UNIVERSIDAD DE CASTILLA -LA MANCHA

ESCUELA TÉCNICA SUPERIOR DE INGENIEROS AGRÓNOMOS Y MONTES

Departamento de Producción Vegetal y Tecnología Agraria
CENTRO REGIONAL DE ESTUDIOS DEL AGUA
Área de Limnología Aplicada e Hidrobiología

Macroalgas bentó nicas de los rı́os de

Castilla - La Mancha.
Bases para el estudio de su potencial
indicador.

Memoria presentada por Laura Monteagudo Canales para optar al grado de Doctor
en Ciencia e Ingeniería Agrarias por la Universidad de Castilla - La Mancha.

Director: José Luis Moreno Alcaraz.

Albacete, 2015
 FINANCIACIÓN

Los trabajos realizados en el desarrollo de esta Tesis Doctoral han sido financiados
por los siguientes convenios y proyectos concedidos por el gobierno regional (Junta
de Comunidades de Castilla-La Mancha):

• PREG01-0016. Convenio específico de colaboración entre la Consejería de Obras

Públicas de la Junta de Comunidades de Castilla-La Mancha y la Universidad de
Castilla-La Mancha en materia de calidad de aguas.
• PREG06-027. Estudio del Estado Ecológico de las masas de agua de CLM:
determinación de caudales ecológicos, estaciones de referencia y cursos naturales.
• PO1109-0190-8090. Uso de la flora acuática como bioindicador de la calidad del
agua de los ecosistemas fluviales de Castilla-La Mancha en aplicación de la
Directiva marco del agua (2000/60/CE).
• PPII10-0271-1349. Macrófitos acuáticos de los ríos de Castilla-La Mancha:
condiciones de referencia, intercalibración de índices de calidad y evaluación del
estado ecológico según la Directiva Marco del Agua (2000/60/CE).
 A mi madre

Brillaste tanto
que sólo con la estela que dejas tras de ti,
iluminas todo mi camino.
Nos vemos al final.
 ÍNDICE

1. RESUMEN ................................................................................................................................. 1
2. INTRODUCCIÓN ........................................................................................................................7
3. OBJETIVOS .............................................................................................................................. 15
3.1. Diversidad y distribución de las macroalgas bentónicas de Castilla-La Mancha ........... 17
3.2. Presiones antropogénicas en el área de estudio ........................................................... 17
3.3. Relación entre la comunidad de cianobacterias y la calidad del agua ........................... 17
4. PLAN DE TRABAJO Y METODOLOGÍA GENERAL ..................................................................... 19
5. ARTÍCULOS CIENTÍFICOS ......................................................................................................... 23
5.1. Lista de géneros de macroalgas fluviales recolectadas en Castilla-La Mancha durante el
período 2001-2014. .................................................................................................................... 25
5.2. Descripción morfológica y ecología de algunas algas consideradas ‘raras’ en los ríos de
Castilla – La Mancha. .................................................................................................................. 41
5.3. Sobre la presencia de Nostochopsis lobata Wood ex Bornet et Flahault en España:
aspectos morfológicos, ecológicos y biogeográficos.................................................................. 53
5.4. Eutrofización fluvial: Comparación del impacto del regadío y secano a través de
diferentes escalas espaciales...................................................................................................... 73
5.5. ¿Son las cianobacterias bentónicas indicadores de presiones antropogénicas en los
sistemas fluviales? ...................................................................................................................... 89
6. DISCUSIÓN GENERAL ............................................................................................................ 103
7. CONCLUSIONES .................................................................................................................... 111
7.1. Diversidad y distribución de las macroalgas bentónicas de Castilla-La Mancha ......... 113
7.2. Presiones antropogénicas en el área de estudio ......................................................... 113
7.3. Relación entre la comunidad de cianobacterias y la calidad del agua ........................ 114
8. BIBLIOGRAFÍA ....................................................................................................................... 117
AGRADECIMIENTOS ...................................................................................................................... 125
 RESUMEN
Nacimiento del Ojuelo, Munera (Albacete)
Fotografía: José Luis Moreno Alcaraz

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1. RESUMEN

Las especies indicadoras ofrecen, simplemente con su presencia, información acerca

de las condiciones físicas y/o químicas del entorno que les rodea. Por tanto, la
evaluación del potencial indicador de las macroalgas fluviales debe partir de una
base sólida de conocimiento acerca de la diversidad de estos organismos y de cómo
responden a las presiones antropogénicas que afectan a la calidad del agua.

En Castilla-La Mancha, la mayoría de estudios sobre algas se han centrado en la

cuenca del Segura, una de las cinco que forman parte de este territorio. En el resto,
los trabajos se centran principalmente en humedales bajo alguna figura de
protección. Por otro lado, del hecho de que la actividad económica principal en la
región sea la agricultura, se puede deducir que este uso de suelo es el primer
causante de la eutrofización en ríos y arroyos. Sin embargo, no todos los tipos de
agricultura afectan con la misma intensidad a las masas de agua, por lo que debería
estudiarse independientemente el impacto de diferentes técnicas agrícolas como el
regadío y el secano, entre otros. Otro factor a tener en cuenta en el análisis de las
presiones es la escala espacial de trabajo, es decir, a qué escala se debe estudiar la
influencia de los usos de suelo en la calidad del agua (p. ej. área total de drenaje,
corredores de un determinado ancho, etc.).

El objetivo final de esta memoria es contribuir a sentar las bases y la metodología

para el estudio del potencial indicador de las macroalgas. Para ello, se parte del
estudio general de la diversidad y distribución de las macroalgas; del análisis de las
presiones responsables de la eutrofización en ríos y arroyos en la región; para
culminar con la aplicación y puesta a prueba de estos conocimientos en el estudio
del potencial indicador de un grupo concreto de macroalgas, las cianobacterias
bentónicas (Figura 1). Como resultado, se han generado cinco artículos científicos
que recogen todas las aportaciones generadas en el desarrollo de esta Tesis
Doctoral.

El primero de ellos (Artículo 5.1.) consiste en un listado de los géneros de

macroalgas bentónicas recolectados en los ríos Castilla-La Mancha durante los

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muestreos llevados a cabo por el Área de Limnología Aplicada e Hidrobiología del
Centro Regional de Estudios del Agua, desde el año 2001 hasta 2014.

En los dos siguientes (Artículos 5.2. y 5.3.), se profundiza en los aspectos

morfológicos, ecológicos y biogeográficos de algunas especies de interés a nivel
europeo, nacional y/o regional que se han detectado en dichos muestreos:
Nostochopsis lobata Wood ex Bornet et Flahault, Batrachospermum atrum (Hudson)
Harvey, Chroothece richteriana Hansg, Oocardium stratum Nägeli, Tetrasporidium
javanicum Möbius e Hydrurus foetidus (Villars) Trevisan.

En el cuarto artículo (5.4.) se recoge el análisis de las presiones antropogénicas que

afectan a la calidad del agua en la región. De este trabajo se desprenden dos ideas
principales: 1) el impacto de la agricultura de regadío en la calidad del agua es
mayor que el de la agricultura de secano; 2) la escala espacial más adecuada para
analizar la presión de los usos de suelo en el área de estudio es la correspondiente a
1km de radio aguas arriba de cada punto de muestreo. Además, se describe la
metodología general para la elección de la escala espacial más adecuada en este tipo
de estudios.

Para finalizar, en el último artículo (5.5.) se aplican las aportaciones de los trabajos
previos para evaluar el potencial indicador de las cianobacterias. El enfoque
estadístico elegido para este trabajo, ha permitido desentrañar la relación
subyacente entre la comunidad de algas y las presiones humanas. Más allá de la
conductividad, que aparece como el factor más determinante en la composición de
especies de estas comunidades, las cianobacterias aportan información acerca de los
niveles de nutrientes en el agua. De entre todas las especies incluidas en este
estudio, cinco han mostrado tener capacidad indicadora de las presiones
antropogénicas y podrían incluirse en los índices de macrófitos empleados en
aplicación de la Directiva Marco del Agua (2000/60/CE): Nostoc verrucosum
Vaucher ex Bornet et Flahault, Phormidium autumnale Gomont, Plectonema
tomasinianum Bornet ex Gomont, Rivularia haematites C. Agardh ex Bornet et
Flahault y Tolypothrix distorta Kützing ex Bornet et Flahault.

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La contribución más importante de esta Tesis Doctoral reside en su interés global, ya
que estas bases pueden ser aplicadas en la evaluación del potencial indicador de
otros grupos de algas (rojas, verdes, etc.) y en otras zonas geográficas.

Figura 1. Estructura general de la presente Tesis Doctoral.

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INTRODUCCIÓN
Río Escabas, Priego (Cuenca)
Fotografía: Laura Monteagudo Canales

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2. INTRODUCCIÓN

La Directiva Marco del Agua (2000/60/CE) (DMA) establece un marco jurídico

comunitario para proteger, regenerar y garantizar el uso sostenible del agua a largo
plazo. En ella se insta a los Estados Miembros a alcanzar el “buen estado” de todas
sus masas de agua antes de diciembre de 2015. El estado de una masa de agua es el
grado de alteración que presenta respecto a sus condiciones naturales y viene
determinado por sus estados químico y ecológico. En cuanto al estado ecológico, los
Estados Miembros deben establecer sistemas de seguimiento de los elementos
biológicos (invertebrados, diatomeas, macrófitos y peces, en el caso de los ríos)
mediante la aplicación de métodos que permitan cuantificar la calidad en las masas
de agua. Dichos métodos utilizan índices o métricas que clasifican finalmente el
estado ecológico de una masa de agua en una de las 5 clases: muy bueno, bueno,
moderado, deficiente y malo (Anexo V, DMA). Como consecuencia de la entrada en
vigor de la DMA, han proliferado este tipo de métricas, siendo más populares las
basadas en invertebrados y diatomeas que las basadas en macrófitos (Ferreira et al.,
2005).

Con el término “macrófitos” se hace referencia a cualquier organismo autótrofo de

tamaño macroscópico que se desarrolla en un ecosistema acuático. Desde un punto
vista sistemático, es un grupo muy amplio y variado, abarcando organismos como
plantas superiores, briófitos y algas (Cirujano et al., 2011). El hecho de que los
índices basados en macrófitos ocupen un segundo plano entre el resto de métricas
parece deberse principalmente a la baja precisión que han mostrado al aplicarse en
diferentes regiones (Demars et al., 2012).

Las algas que desarrollan talos macroscópicos se denominan comúnmente

“macroalgas” y son organismos exclusivamente acuáticos, íntimamente ligados a las
condiciones ambientales del agua. A pesar de que la DMA las propone como
indicador biológico del estado ecológico junto con el resto de macrófitos (Anexo V,
DMA), la mayoría de macroalgas suelen estar poco representadas en los índices de
macrófitos. Sin embargo, recientemente se está corrigiendo esta tendencia. A nivel
nacional, se ha realizado un esfuerzo en generar diversos índices de macrófitos que

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incluyen las macroalgas en sus listados de taxones indicadores, con el objetivo de ser
aplicados en la evaluación del estado ecológico de los ríos peninsulares (Moreno et
al., 2006, 2008; Suárez et al., 2005; Flor-Arnau et al., 2015).

Hay dos factores que pueden estar detrás de esta discriminación: la falta de
conocimiento sobre las respuestas de estos organismos a las presiones
antropogénicas (Thacker and Paul, 2001) y la complejidad de la identificación de
algunos taxones a nivel de especie (Marquardt & Palinska, 2007). Cabe pensar,
entonces, que es posible perfeccionar los índices de macrófitos identificando, e
incorporando en ellos, aquellas especies de macroalgas que cumplan con los
requisitos que debe cumplir un buen bioindicador. Estos serían, según Bellinger y
Sigee (2010): aportar información sobre las condiciones físicas y/o químicas del
entorno que les rodea, tener un rango ecológico más bien estrecho, una distribución
geográfica amplia y que su identificación taxonómica sea fiable.

Como primer paso para identificar estos bioindicadores entre las macroalgas, es
fundamental ahondar en el conocimiento de su diversidad y ecología, así como de las
presiones que afectan a la calidad del agua que habitan. En lo que respecta a la flora
algal, en los últimos años se ha ampliado el conocimiento sobre su diversidad y
distribución geográfica como consecuencia de la aplicación de la DMA. Castilla-La
Mancha cuenta con un escenario hidrogeológico muy diverso, estando bien
representados los ríos silíceos y calcáreos, tramos altos y medios, climas húmedos y
secos, ríos permanentes y temporales (Moreno et al., 2006). En estos ríos
temporales, muy comunes en la zona mediterránea, es difícil que puedan
desarrollarse plantas vasculares y briófitos verdaderamente acuáticos ya que gran
parte del año están secos (Dodkins et al., 2012). Por el contrario, las algas sí tienen la
capacidad de desarrollarse en estos cortos periodos de tiempo y además soportan
condiciones de sequía siendo capaces de restablecerse rápidamente al restaurarse el
caudal (Romani and Sabater, 1997; Robson, 2000, Robson et al., 2008).

Esta diversidad de ecosistemas acuáticos en la región favorece la existencia de una

alta biodiversidad de la flora acuática en general, y de la flora algal en particular. Sin
embargo, los estudios sobre algas en Castilla-La Mancha son muy escasos. De las

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cinco cuencas principales que incluye la región, Tajo, Guadiana, Júcar, Guadalquivir y
Segura, ésta última es la más intensamente estudiada desde el punto de vista de las
algas fluviales (Aboal y Llimona, 1985; Aboal, 1988a, b, c; Aboal, 1989a, b, c; Sabater
et al., 1989). En cuanto al resto de cuencas, las publicaciones se centran
principalmente en los humedales de la región, destacando los trabajos de Aboal
(1996), Álvarez-Cobelas (2007) y Cirujano y Medina (2002).

Con el objeto de contribuir al conocimiento de las macroalgas en la región, el Área de

Limnología Aplicada e Hidrobiología del Centro Regional de Estudios del Agua ha
venido realizando campañas extensivas de muestreo en los ríos de la región durante
el período 2001-2014. Estos datos han constituido la base florística y ecológica en el
desarrollo de la presente Tesis Doctoral. Desde un punto de vista aplicado, destaca la
aportación por dicho equipo de un índice de macrófitos para la evaluación del
estado ecológico de los ríos (IVAM, Índice de Vegetación Acuática Macroscópica;
Moreno et al., 2006). Este índice recoge un listado de 36 géneros de macroalgas
recolectados en la región hasta mitad de la década pasada, constituyendo un primer
referente a escala regional.

En cuanto a las presiones antropogénicas que sufren los ecosistemas acuáticos, uno
de los impactos que producen mayor detrimento en la calidad del agua es la
eutrofización, consecuencia de la transformación de terrenos naturales a sistemas
dominados por la actividad humana (Lund, 1967; Omernik et al., 1981; Smith, 2003).
La eutrofización provoca cambios en la composición de especies y la proliferación de
algas filamentosas, lo que conlleva el descenso del oxígeno disuelto, el
empeoramiento de la calidad del agua y la pérdida de biodiversidad (Carpenter et
al., 1998; Quinn, 1991; Smith et al., 1999).

La relación entre la eutrofización y las especies bioindicadoras puede analizarse a

través de la concentración de nutrientes del agua, una medida directa, aunque
puntual y esporádica; mientras que el análisis de los usos de suelo, representa una
forma indirecta aunque posiblemente más precisa, ya que refleja el escenario real
que genera el impacto a más largo plazo. Para analizar la relación entre usos de
suelo y calidad del agua de forma adecuada, debe tenerse en cuenta que intervienen

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numerosos factores a través de diferentes escalas espaciales y temporales (Frissell
et al., 1986). De ahí que puedan surgir algunas cuestiones como: (1) qué clases de
usos de suelo deben contabilizarse; (2) qué escala espacial de trabajo sería la más
apropiada para ello.

En cuanto a la primera cuestión, los usos de suelo más comúnmente estudiados

como causa de eutrofización de ríos y arroyos a nivel global, han sido el urbano y el
agrícola, ya que son fuentes de contaminación puntual y difusa, respectivamente
(Osborne and Wiley, 1988; Townsend et al., 1997; Schiller et al., 2008). En el caso de
Castilla-La Mancha, prácticamente la mitad de su superficie está destinada a la
agricultura (Ministerio de Agricultura, Alimentación y Medio Ambiente, 2014), por
lo que la contaminación difusa es muy relevante. Sin embargo, no todos los tipos de
agricultura tienen el mismo impacto. Existen trabajos previos en los que se
comparan diferentes actividades agrícolas. Por ejemplo, Johnson et al. (1997) que
compararon el cultivo en hileras con la agricultura de conservación (sin arado) o
Lassaletta et al. (2009) que compararon entre cultivos permanentes, terreno arable
y zonas mixtas en España. Según el Instituto Nacional de Estadística (www.ine.es) el
regadío es más productivo que el secano debido al aporte artificial de agua y
fertilizantes a los cultivos. Mediante esta técnica, aumenta el drenaje de compuestos
de nitrógeno a las aguas superficiales, lo que la convierte en la principal causa de
eutrofización de ríos en España (Cavero et al., 2003; Berzas et al., 2004; Álvarez-
Cobelas et al., 2010). Por tanto, aunque en Castilla-La Mancha el regadío supone
aproximadamente el 12% del total del suelo agrícola (Ministerio de Agricultura,
Alimentación y Medio Ambiente, 2014), parece lógico diferenciarlo de otros tipos de
agricultura más extensos pero con menor impacto potencial. Por todo ello, en el
desarrollo de los trabajos que componen esta memoria se ha dividido el uso agrícola
en tres subclases: regadío, secano y agricultura de bajo impacto (zonas agrícolas con
una alta densidad de vegetación natural).

En cuanto a la segunda cuestión, hay autores que sugieren que debe emplearse la
escala de cuenca de drenaje (Figura 2.a.) (Omernik et al., 1981; Richards y Host,
1994; Roth et al., 1996) ya que consideran que todo impacto aguas arriba de un
punto tiene efecto en él; mientras otros argumentan que los usos de suelo más

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cercanos tienen mayor influencia y, por tanto, deben emplearse escalas locales,
como los corredores o zonas de influencia (Figura 2.b. y 2.c., respectivamente)
(Harding et al., 1998; Nerbonne y Vondracek, 2001).

Figura 2. Representación de las diferentes escalas espaciales empleadas en el estudio de la

influencia de los usos de suelo en la calidad del agua. a: subcuenca de drenaje; b: corredor
longitudinal; c: zona de influencia (buffer radial).

En estudios más recientes, algunos autores optan por hacer una evaluación previa a
través de varias escalas espaciales para determinar cuál es la más apropiada para el
estudio en cuestión (p. ej. Chang, 2008; Tran et al., 2010). Este marco de trabajo
‘multi-escala’ es el que se ha considerado en el desarrollo de esta tesis. Así, se
contabilizaron los usos de suelo que afectan a cada punto a través de diferentes
escalas espaciales, de forma que los datos obtenidos variaron de una escala a otra. El
análisis profundo de estos datos ha permitido detectar cuál es la escala más
apropiada para el estudio del potencial indicador de las macroalgas bentónicas.

De entre todas las algas, las cianobacterias (o algas verde-azules) destacan por ser
un grupo muy diverso, extendido y cuyas características fisiológicas exclusivas
hacen que se puedan ver influenciadas por los nutrientes de manera distinta al resto
de algas. Estos organismos son capaces de fijar nitrógeno atmosférico, lo que les
permite vivir en condiciones de escasez de compuestos de nitrógeno disueltos. Por
ello, algunos autores sugieren que su dependencia de nitrógeno en comparación con
otros grupos de algas puede ser mucho menor (Larkum et al., 1988). Además,
también existen diferencias entre cianobacterias con y sin heterocitos. Las primeras,
son capaces de fijar nitrógeno atmosférico (N 2 ) en condiciones aerobias, mientras

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que las segundas tienen esta capacidad limitada a condiciones anaerobias y/o de
oscuridad (Potts, 1979; Lee, 2008). Loza et al. (2014) señalan que esta ventaja
ecofisiológica puede ser el motivo de que las cianobacterias con heterocitos sean
dominantes en ambientes pobres en compuestos de nitrógeno, y que las
cianobacterias sin heterocitos tengan preferencia por zonas con altos niveles de los
mismos. Con respecto al fósforo, algunas especies, como las del género Rivularia
pueden sobrevivir en concentraciones limitantes de fósforo gracias a la actividad
fosfatasa, la cual puede utilizarse como un buen indicador de condiciones
oligotróficas (Mateo et al., 2010). Con todo ello, hay diversos estudios en todo el
mundo que las relacionan con actividades humanas causantes de eutrofización en
sistemas acuáticos (p.ej. Johansson, 1982; Jafari y Gunale, 2006; Parikh et al., 2006).

Todos estos factores ponen en evidencia que las cianobacterias son un grupo de
organismos muy diverso, en el que encontramos especies propias tanto de sitios
impactados como de sitios limpios. Por tanto, cabe esperar que algunas de estas
especies sean buenos indicadores que puedan tenerse en cuenta en el desarrollo de
índices biológicos de la calidad de agua. Para identificarlas, es esencial analizar la
relación entre la comunidad de cianobacterias y el entorno, así como aplicar los
requisitos que debe cumplir un buen bioindicador.

A través del caso concreto de las cianobacterias, este trabajo pretende sentar las
bases generales del estudio del potencial indicador de las macroalgas en un marco
de trabajo adecuado, de forma que sean aplicables al estudio de otros grupos de
algas en el futuro.

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 OBJETIVOS
Arroyo Alarconcillo, Ossa de Montiel (Albacete)
Fotografía: José Luis Moreno Alcaraz

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3. OBJETIVOS

En este capítulo quedan recogidos los objetivos generales de esta Tesis Doctoral,
agrupados según las líneas de trabajo planteadas en la estructura de la misma
(Figura 1). Los objetivos específicos de cada publicación pueden ser consultados en
su correspondiente capítulo.

3.1. Diversidad y distribución de las macroalgas bentónicas de Castilla-La

Mancha:

 Conocer la diversidad y la distribución de las macroalgas bentónicas de

Castilla-La Mancha (Artículos 5.1., 5.2. y 5.3.).
 Describir la morfología y ecología de algunas especies consideradas ‘raras’ a
nivel europeo o que suponen primeras citas en la región (Artículos 5.2. y
5.3.).
 Revisar y comparar los aspectos morfológicos, ecológicos y la distribución
geográfica de Nostochopsis lobata Wood ex Bornet et Flahault (Artículo 5.3.).

3.2. Presiones antropogénicas en el área de estudio:

 Demostrar que la escala espacial influye en los resultados de los estudios que
relacionan los usos de suelo con la calidad del agua (Artículo 5.4.).
 Determinar qué tipo de agricultura es responsable, en mayor medida, de la
eutrofización de ríos y arroyos en la región, así como cuantificar su umbral de
presión (Artículo 5.4.).
 Proponer un protocolo general que ayude a la elección de la escala espacial
más adecuada en estudios relacionados con el impacto de la contaminación
difusa (Artículo 5.4.).

3.3. Relación entre la comunidad de cianobacterias y la calidad del agua:

 Analizar las variables ambientales que determinan la comunidad de

cianobacterias bentónicas (Artículo 5.5.).

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 Examinar las diferencias entre cianobacterias con y sin heterocitos (Artículo
5.5.).
 Identificar qué especies son útiles como indicadoras de presiones
antropogénicas en la zona de estudio (Artículo 5.5.).

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PLAN DE TRABAJO Y
METODOLOGÍA
Río Sorbe, Galve de Sorbe (Guadalajara)
Fotografía: Laura Monteagudo Canales

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4. PLAN DE TRABAJO Y METODOLOGÍA GENERAL

El trabajo desarrollado durante estos años puede agruparse en tres fases o líneas de
estudio (Figura 3).

Figura 3. Esquema del plan de trabajo. Se relacionan los aspectos metodológicos y objetivos
principales de cada una de las tres fases.

Las tres fases se abordaron siguiendo un orden cronológico. La primera fase

comenzó en 2001, cuando se constituyó el equipo de Limnología Aplicada e
Hidrobiología del Centro Regional de Estudios del Agua, años antes del comienzo de
este proyecto doctoral. Los datos reunidos hasta el momento fueron incluidos y
ampliados durante el desarrollo de la misma, hasta el año 2014. Durante los

21
muestreos se procedió a la recogida de las colonias de macroalgas en tramos de 100
m recorridos en zigzag, cubriendo la mayor variedad de hábitats posible. A su vez,
también se tomaron muestras de agua para su análisis en laboratorio. Todas las
macroalgas recogidas se identificaron, como mínimo, a nivel de género y se procedió
a su conservación en formaldehido 3%. Con parte del material, se realizaron
preparaciones permanentes con glicero-gelatina, pliegues o sobres en seco, y viales
con gel de sílice. La colección se encuentra almacenada en el citado centro de
investigación.

La fase referente al análisis de las presiones antropogénicas comenzó en 2010 y se

llevó a cabo paralelamente a los muestreos de campo. En ella se analizaron capas
vectoriales de usos de suelo (Corine Land Cover) y se establecieron las clases de
usos de suelo a analizar: forestal, uso urbano, cultivos de regadío, cultivos de secano
y terrenos agrícolas considerados de bajo impacto ambiental (pastos o cultivos
abandonados con alta densidad de vegetación natural). El análisis del impacto de
estas clases de suelo se analizó a través de diferentes escalas: área de drenaje,
corredores, y zonas de influencia de 1 y 5 km de radio aguas arriba de los puntos de
muestreo.

La última, fue la fase de aplicación de los conocimientos adquiridos y metodologías

descritas en fases anteriores. Para ello se seleccionó un subgrupo de macroalgas, las
cianobacterias, con buena representación territorial, y fueron identificadas a nivel
de especie. Con este grupo ‘piloto’ se evaluó el efecto de diferentes variables
ambientales en la composición de especies de la comunidad. Se aplicaron diferentes
enfoques estadísticos para desentrañar las relaciones más sutiles, ya que a veces
quedan enmascaradas bajo la influencia de factores que actúan a una mayor escala
(p. ej conductividad).

La descripción detallada de la metodología empleada en cada procedimiento puede

consultarse en la sección de material y métodos de los artículos en cuestión
(Capítulo 5).

22
 ARTÍCULOS
CIENTÍFICOS
Rivularia biasolettiana
Fotografía: José Luis Moreno Alcaraz

23
24
5. ARTÍCULOS CIENTÍFICOS

5.1. Lista de géneros de macroalgas fluviales recolectadas en Castilla-La

Mancha durante el período 2001-2014.

Título original: Lista de géneros de macroalgas fluviales

recolectadas en Castilla-La Mancha durante el
período 2001-2014
Autores: José Luis Moreno y Laura Monteagudo
Estado: En revisión
Fecha: 2015

RESUMEN

El presente trabajo recoge la lista de géneros de macroalgas fluviales de Castilla-La

Mancha, resultado del muestreo de 178 estaciones situadas en ríos y arroyos a lo
largo de la región, durante el periodo comprendido entre los años 2001 y 2014. Esta
lista incluye un total de 64 géneros, todos ellos capaces de formar colonias o talos de
tamaño macroscópico. También se presenta la localización geográfica precisa de los
tramos fluviales donde se han recolectado, con indicación de la provincia y cuenca
hidrológica a la que pertenecen. Este listado supone un primer acercamiento a la
comunidad de macroalgas fluviales de la región, y puede servir como base para
futuros estudios de calidad del agua, ficológicos y ecológicos.

25
26
Lista de géneros de macroalgas fluviales recolectadas en Castilla-La
Mancha durante el período 2001-2014
José Luis Moreno y Laura Monteagudo
Universidad de Castilla-La Mancha, Centro Regional de Estudios del Agua (CREA)
Crta. de Las Peñas km. 3, Albacete 02071, España.

RESUMEN

El presente trabajo recoge la lista de géneros de macroalgas fluviales de Castilla-La

Mancha, resultado del muestreo de 178 estaciones situadas en ríos y arroyos a lo largo
de la región, durante el periodo comprendido entre los años 2001y 2014. Esta lista
incluye un total de 64 géneros, todos ellos capaces de formar colonias o talos de tamaño
macroscópico. También se presenta la localización geográfica precisa de los tramos
fluviales donde se han recolectado, con indicación de la provincia y cuenca hidrológica a
la que pertenecen. Este listado supone un primer acercamiento a la comunidad de
macroalgas fluviales de la región, y puede servir como base para futuros estudios de
calidad del agua, ficológicos y ecológicos.

1. INTRODUCCION posible papel como bioindicadores de

Castilla-La Mancha incluye en su territorio
cinco grandes cuencas hidrográficas: Tajo,
Guadiana, Júcar, Segura y Guadalquivir. Es
una de las regiones más extensas del país y
cuenta con un escenario hidrogeológico muy
diverso, estando bien representados los ríos
silíceos y calcáreos, tramos altos y medios,
climas húmedos y secos, ríos permanentes y
temporales (Moreno et al., 2006). Ello se
traduce en una alta biodiversidad de la flora
acuática en general y, de la flora algal en
particular. Sin embargo, son muy escasos los
estudios sobre algas que se han llevado a
cabo en la región. Entre ellos, cabe destacar
los trabajos realizados por Aboal (1988a;
1988b; 1989a; 1989b) y Sabater et al.
(1989), centrados fundamentalmente en la
cuenca del Segura, cuya cabecera se localiza
en la provincia de Albacete.
Como consecuencia de la entrada en vigor
de la Directiva Marco del Agua
(2000/60/CE), en adelante DMA, se ha
incrementado el interés por las algas y su
calidad del agua. Las algas que desarrollan
talos macroscópicos, denominadas
comúnmente “macroalgas”, forman parte,
junto con briófitos y plantas vasculares, del
indicador biológico denominado
“macrófitos” (Anexo V, DMA). Por ello, las
macroalgas son habitualmente incluidas en
los índices de macrófitos, índices que son
utilizados en la evaluación del estado
ecológico de los ríos mediante la flora
acuática (p.ej. Moreno et al., 2006a; Flor-
Arnau et al., 2015). Por tanto, ampliar la
información disponible sobre la
distribución, diversidad y ecología de las
algas fluviales, es fundamental para poder
desarrollar herramientas eficaces en
materia de bioindicación.
Este trabajo pretende contribuir al
conocimiento de la diversidad de
macroalgas en la región, proporcionando
una lista de los géneros detectados durante
un periodo de trece años (2001-2014), junto
con su localización geográfica precisa. La
información aportada también es de interés

27
a nivel peninsular, dado que son muy
escasos los estudios extensivos sobre
macroalgas fluviales publicados hasta el
momento. Por último, algunas especies con
mayor interés, bien por su rareza o bien por
ser primeras citas nacionales y/o
regionales, se han tratado de forma más
extensa en dos trabajos previos (Moreno et
al., 2012; 2013).
2. MATERIAL Y MÉTODOS
2.1. Área de estudio
El régimen climático al que pertenece la
región es el mediterráneo-continentalizado,
marcado por inviernos fríos y veranos
calurosos con fuertes oscilaciones térmicas.
En cuanto a las precipitaciones,
encontramos desde zonas áridas como la
llanura manchega y el sureste de la región
con valores inferiores a los 300mm al año,
hasta zonas montañosas donde se superan
los 1000mm (Fernández, 2000).
Geológicamente, se diferencian tres grandes
zonas: la zona oeste de naturaleza silícea,
formada por esquistos, gneises, pizarras y
granitos; la llanura central sedimentaria,
abundante en calizas, arcillas y yesos; y la
zona este, caracterizada por la presencia de
rocas calcáreas como calizas, dolomías,
margas, arcillas y conglomerados (Porras
Martín et al., 1985). Esta diversidad
climática, geológica y geográfica ha
originado también una red hidrológica
formada por ríos y arroyos de diversas
tipologías (Moreno et al., 2006b).
2.2. Diseño del muestreo y análisis del
material
El presente estudio fue llevado a cabo en
ríos y arroyos de Castilla-La Mancha, entre
los años 2001-2014. En total, se
recolectaron macroalgas en 179 puntos de
muestreo (Figura 1), abarcando todo el
gradiente climático y litológico presente en
la región. En cada estación de muestreo, se
recogieron a mano o con navaja las
macroalgas detectadas, es decir, aquellas
algas capaces de desarrollar talos de tamaño
macroscópico, visibles simple vista. Estas
28
presentaron diversas morfologías
(laminares, globulares, mechones, madejas
filamentosas, masas gelatinosas) y formas
de vida (adheridas o tapizando diferentes
sustratos, enraizadas en sedimentos,
flotantes).
Los muestreos se realizaron recorriendo
tramos de 100 m en zigzag de orilla a orilla
y río arriba, cubriendo toda la variedad de
microhábitats y sustratos presentes. El
material recogido fue refrigerado y
trasladado al laboratorio para su
identificación. Las obras empleadas para
ello fueron principalmente las siguientes:
Bornet and Flahault (1887), Geitler (1932),
Bourrelly (1957; 1990), Desikachary
(1959), Aboal (1988a; 1988b; 1989a;
1989b), Komárek and Anagnostidis (2005)
Eloranta and Kwandrans (2007), Ettl and
Gartner (2009) y Eloranta (2011). El nivel
taxonómico alcanzado ha sido el de género,
ya que es el alcanzado habitualmente en la
aplicación de los índices de macrófitos
utilizados en la evaluación del estado
ecológico de los ríos. Por otra parte,
actualmente existe una gran inestabilidad
taxonómica en numerosos grupos de algas a
nivel específico (e incluso genérico), debido
a la reciente aplicación de nuevas técnicas
de análisis genéticos.
Posteriormente, el material fue fijado con
formaldehido 3% para su conservación. En
los casos de interés, también se realizaron
preparaciones permanentes con glicero-
gelatina, pliegues o sobres en seco, y viales
con gel de sílice. La colección se encuentra
almacenada en el Centro Regional de
Estudios del Agua (Universidad de Castilla-
La Mancha), Laboratorio de Limnología
Aplicada e Hidrobiología, Albacete.
2.3. Información geográfica
La localización de los puntos de muestreo se
realizó en el entorno ArcGis 9, generando un
mapa con cuadrícula superpuesta de
coordenadas UTM. El listado de géneros
incluye información sobre las provincias de
la región donde fueron recolectados,
mientras que en el listado de puntos de
muestreo se incluye el municipio y la cuenca
hidrológica a la que pertenecen.
Figura 1. Mapa de la zona de estudio dividida en cuadrículas UTM. Las líneas blancas marcan los límites
provinciales (Ab, Albacete; Cu, Cuenca; CR, Ciudad Real; Gu, Guadalajara; To, Toledo).

3. RESULTADOS

El listado reúne un total de 64 géneros de macroalgas fluviales detectados en Castilla-La

Mancha durante el periodo de estudio. Entre ellos aparecen; 27 representantes del filo
Cyanophyta (=Cyanobacteria) (algas verdeazules); 1 del filo Bacillariophyta (diatomeas); 3
del filo Ochrophyta (algas ocres); 11 del filo Rhodophyta (algas rojas); 15 del filo
Chlorophyta y 7 del filo Charophyta ambos pertenecientes al grupo de las llamadas algas
verdes. Según la nomenclatura actualizada proporcionada por el proyecto www.algaebase.org.

Anabaena Bory de Saint-Vincent ex Bornet & Flahault - Ab, CR, To - 13, 17, 18, 19, 20,
21

Audouinella Bory de Saint-Vincent - Ab, CR, Cu, Gu - 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,
14, 15, 16

Bangia Lyngbye - Cu, Gu - 134, 140, 166, 173

29
Batrachospermum Roth - Ab, CR, Cu, Gu - 1, 3, 4, 5, 6, 7, 8, 13, 14, 16, 30, 40, 41, 42, 47,
56, 57, 59, 68, 81, 86, 87, 88, 89, 90, 91, 92, 99, 100, 103, 105, 109, 112, 118, 122,
125, 127, 128, 134, 177

Bulbochaete C.Agardh - Cu, Gu - 111, 128

Calothrix C.Agardh ex É.Bornet & C.Flahault – CR - 18

Chaetophora F.Schrank - Ab, CR, Cu, Gu - 112, 119, 86, 15, 111, 8, 4, 68, 42, 134, 5, 105,
11, 124, 155, 66, 82, 102, 108, 87, 89, 151, 79, 18, 148

Chamaesiphon A.Braun – Gu - 146, 148, 151, 153, 178, 179

Chara Linnaeus - Ab, CR, Cu, Gu, To - 3, 4, 6, 7, 13, 14, 15, 16, 17, 22, 29, 36, 42, 44, 45,
46, 53, 54, 56, 59, 64, 65, 77, 79, 85, 86, 87, 88, 102, 105, 108, 109, 112, 113, 116,
118, 119, 125, 128, 129, 134, 142, 144, 147, 155, 161, 166, 169, 173

Chroodactylon Hansgirg – Ab - 16

Chroothece Hansgirg - Ab, Gu - 7, 14, 118

Cladophora Kützing - Ab, CR, Cu, Gu, To - 1, 3, 4, 6, 7, 9, 10, 12, 13, 14, 16, 18, 21, 22, 23,
24, 25, 28, 30, 33, 35, 36, 37, 39, 40, 41, 44, 46, 50, 51, 52, 53, 54, 55, 56, 58, 61, 62,
63, 64, 65, 66, 68, 75, 77, 78, 79, 83, 85, 88, 89, 90, 91, 92, 93, 94, 95, 97, 98, 100,
101, 102, 106, 107, 108, 109, 110, 112, 113, 116, 117, 118, 120, 121, 123, 124, 125,
127, 128, 133, 134, 135, 136, 137, 138, 139, 140, 141, 146, 150, 154, 155, 156, 157,
160, 161, 163, 166, 168, 169, 171, 173, 174

Coleodesmium A.Borzì ex L.Geitler - CR, Gu - 2, 141, 148, 153

Compsopogon Montagne – Ab - 7

Cylindrospermum - F.T.Kützing ex É.Bornet & C.Flahault - Ab, CR - 7, 73, 108

Draparnaldia Bory de Saint-Vincent - Ab, CR, Cu, Gu - 2, 10, 11, 42, 66, 81, 82, 105, 146,
148, 149, 151, 152, 179

Enteromorpha Link – Ab - 107

Geitlerinema (Anagnostidis & Komárek) Anagnostidis – Ab - 54

Gloeocapsa Kützing - Ab, Gu - 94, 154

Gloeotrichia J.Agardh ex Bornet & Flahault – CR - 73

Heteroleibleinia (L.Geitler) L.Hoffmann - CR, Gu - 83, 179

Hildenbrandia Nardo - Ab, Cu, Gu, To - 4, 7, 57, 116, 133, 140, 146

30
Homeothrix (Thuret ex Bornet & Flahault) Kirchner – Gu - 179

Hyalotheca Ehrenberg ex Ralfs – Cu - 123

Hydrococcus Kützing – Ab - 7

Hydrocoleum Kützing ex Gomont – Ab – 87

Hydrodictyon Roth – CR – To - 18, 19, 20, 27, 32, 79, 84, 114, 132

Hydrurus C.Agardh – Gu - 141, 179

Klebsormidium P.C.Silva, Mattox & W.H.Blackwell – Ab - 89

Kyliniella Skuja – Ab - 108

Lemanea Bory - CR, Cu, Gu, To - 2, 5, 11, 66, 71, 79, 80, 82, 116, 125, 133, 141, 143, 148,
151, 153, 155, 178, 179

Leptolyngbya Anagnostidis & Komárek – To - 133, 161

Lyngbya C.Agardh ex Gomont - Ab, To - 13, 164, 168

Melosira C.Agardh - Ab, CR, Cu, Gu - 10, 18, 39, 55, 95, 107, 140, 149, 153, 154, 177

Microcoleus Desmazières ex Gomont - Cu, Gu, To - 19, 134, 140, 143

Microspora Thuret - Ab, CR, Cu, Gu, To - 1, 8, 14, 16, 17, 19, 40, 41, 42, 84, 85, 89, 114,
127, 128, 133, 172

Monostroma Thuret - Cu, Gu, CR - 83, 58, 124

Mougeotia C.Agardh - Ab, CR, Cu, Gu, To - 13, 14, 27, 34, 47, 54, 56, 66, 76, 145,
149, 153, 166, 169, 175

Nitella C.Agardh - Ab, CR - 13, 17, 69, 70, 76

Nostoc Vaucher ex Bornet & Flahault - Ab, CR, Cu, Gu, To - 1, 2, 3, 5, 6, 7, 11, 12, 15, 18,
19, 22, 28, 51, 54, 58, 62, 64, 66, 79, 84, 87, 89, 90, 95, 97, 99, 104, 108, 109, 116,
123, 125, 129, 141, 148, 151, 153, 166, 169, 178, 179

Nostochopsis H.C.Wood ex É.Bornet & C.Flahault – CR - 18

Oedogonium Link ex Hirn - Ab, CR, Cu, Gu, To - 5, 7, 8, 9, 11, 12, 13, 14, 15, 16, 17, 18,
19, 21, 34, 41, 42, 47, 50, 51, 56, 57, 64, 66, 70, 71, 76, 78, 79, 80, 82, 84, 85, 87, 88,
89, 91, 110, 111, 123, 128, 135, 139, 154, 155, 161, 163, 166, 168, 169, 177

Oocardium Nägeli – Ab - 14

31
Oscillatoria Vaucher ex Gomont - Ab, CR, Cu, Gu, To - 25, 44, 46, 72, 93, 123, 150, 164,
173, 175

Paralemanea (P.C.Silva) Vis & Sheath – To - 161

Phormidium Kützing ex Gomont - Ab, CR, Cu, Gu, To - 1, 3, 4, 6, 7, 10, 11, 12, 13, 14, 18,
21, 22, 27, 28, 29, 30, 31, 32, 33, 34, 36, 37, 38, 39, 40, 41, 42, 43, 48, 50, 51, 52, 54,
56, 57, 64, 67, 68, 70, 72, 81, 83, 87, 88, 89, 91, 92, 94, 96, 97, 98, 100, 102, 103, 106,
108, 116, 118, 125, 133, 139, 140, 142, 143, 148, 153, 155, 159, 161, 164, 177, 178,
179

Plectonema Thuret ex Gomont - Ab, Cu, Gu - 1, 13, 14, 30, 42, 56, 64, 87, 89, 93, 100, 104,
175

Pseudanabaena Lauterborn - Ab, Cu, Gu, To - 16, 21, 47, 159

Rhizoclonium Kützing - Ab, CR, Cu, Gu, To - 9, 21, 23, 25, 30, 32, 37, 39, 110, 114, 115,
120, 123, 126, 132, 135, 137, 145, 147, 157, 168

Rivularia C.Agardh ex É.Bornet & C.Flahault - Ab, CR, Cu, Gu - 3, 5, 13, 14, 34, 42, 43,
44, 45, 47, 53, 59, 60, 66, 68, 85, 86, 87, 88, 89, 91, 92, 99, 101, 102, 103, 104, 105,
108, 109, 112, 128, 129, 134, 142, 155, 166, 169, 173, 175

Schizothrix F.T.Kützing ex M.Gomont - Ab, Cu, Gu - 14, 15, 16, 56, 57, 88, 90, 92, 96,
100, 109, 134, 175

Scytonema C.Agardh ex É.Bornet & C.Flahault - Ab, CR, Cu, Gu - 2, 6, 34, 42, 53, 85, 88,
91, 102, 104, 128, 141, 153, 170

Spirogyra Link in Nees - Ab, CR, Cu, Gu, To - 1, 2, 3, 5, 7, 9, 10, 11, 12, 13, 14, 15, 17, 18,
19, 20, 21, 23, 24, 26, 27, 32, 33, 34, 36, 38, 39, 41, 42, 43, 45, 47, 51, 54, 56, 57, 58,
59, 60, 71, 75, 76, 77, 78, 79, 80, 81, 82, 84, 87, 90, 91, 93, 96, 97, 99, 100, 101, 102,
103, 104, 105, 108, 109, 113, 114, 115, 116, 119, 123, 124, 128, 131, 133, 135, 141,
142, 145, 147, 148, 149, 150, 151, 152, 153, 155, 161, 162, 166, 168, 169, 172, 173,
177, 178

Stigeoclonium Kützing - Ab, CR, Gu, To - 12, 16, 18, 42, 78, 95, 107, 116, 133, 135, 168,
171, 172, 178

Tetraspora Link ex Desvaux - Ab, CR, Cu, Gu, To - 18, 41, 68, 70, 71, 79, 82, 97, 149, 151,
162, 179

Tetrasporidium Möbius - CR, Gu - 2, 18, 70, 71, 149, 151

32
Thorea Bory – Ab - 7, 49

Tolypella (A.Braun) A.Braun – Gu - 142

Tolypothrix F.T.Kützing ex É.Bornet & C.Flahault - Ab, Cu, Gu - 6, 7, 11, 13, 42, 54, 56,
66, 86, 88, 92, 93, 100, 101, 103, 108, 116, 118, 122

Tribonema Derbès & Solier - Gu, To - 19, 21, 149, 153

Trichocoleus K.Anagnostidis – Gu - 175

Ulothrix Kützing - Ab, Cu, Gu - 1, 4, 11, 95, 148, 153

Vaucheria A.P.de Candolle - Ab, CR, Cu, Gu, To - 3, 6, 7, 9, 10, 12, 29, 30, 33, 36, 39, 41,
51, 52, 54, 55, 58, 62, 63, 68, 75, 80, 83, 89, 93, 94, 95, 97, 106, 110, 114, 120, 126,
127, 130, 138, 140, 143, 154, 157, 162, 176

Zygnema C.Agardh - Ab, CR, Cu, Gu - 5, 13, 14, 15, 17, 18, 26, 27, 32, 41, 42, 47, 53, 54,
56, 59, 60, 66, 71, 73, 76, 80, 82, 85, 87, 88, 91, 97, 99, 101, 103, 104, 105, 108, 109,
111, 113, 116, 119, 128, 141, 142, 144, 147, 153, 166, 169, 172

Agradecimientos

Este estudio ha sido realizado en el Centro Regional de Estudios del Agua (Universidad de
Castilla-La Mancha, Albacete) y financiado por los siguientes convenios y proyectos
concedidos por el gobierno regional (Junta de Comunidades de Castilla-La Mancha):
PREG01-0016, PREG06-027, PO1109-0190-8090 y PPII10-0271-1349.

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2012. On the presence of Nostochopsis
lobata Wood ex Bornet et Flahault in
Spain: morphological, ecological and
biogeographical aspects. Nova
Hedwigia 95, 373–390.
Moreno, J.L., Monteagudo, L. and Aboal, M.,
2013. Morphological description and
ecology of some rare macroalgae in
south-central Spanish rivers (Castilla-
La Mancha Region). An. Jardín
Botánico Madr. 70, 81–90.
Porras Martín, J., Ruiz, C., Fernández, J.A.,
Gómez de las Heras J. y Fabregat, V.
1985. Síntesis Hidrogeológica de
Castilla-La Mancha. Instituto
Geológico y Minero de España,
Madrid.
Sabater, S., Aboal, M. and Cambra, J. 1989.
New Rhodophyceae records for the NE
and SE Spanish continental waters.
Limnetica 5, 93–100.

ANEXO

Listado y localización geográfica de los puntos o estaciones de muestreo, expresada de la

siguiente manera: Número de estación: Nombre del cuerpo de agua (Cuenca hidrográfica)
Localidad, Municipio; Cuadrícula UTM 10x10 km; Altitud (m)

1: Río Horcajo (Guadalquivir) en El Horcajo, Alcaraz; 30SWH48; 1017 m

2: Río Estena (Guadiana) en Rocigalgo, Hontanar; 30SUJ68; 816 m
3: Río Cabriel (Júcar) en Salvacañete; 30TXK24; 1164 m
4: Río Guadazaón (Júcar) en Yemeda; 30SXK00; 852 m
5: Río Huécar (Júcar) en Palomera; 30TWK83; 1104 m
6: Río Júcar (Júcar) en Cuasiermas, Albacete; 30SXJ03; 623 m
7: Río Júcar (Júcar) en Las Mariquillas, Valdeganga; 30SXJ03; 607 m

34
8: Río Pesebre (Júcar) en Pesebre, Peñascosa; 30SWH58; 1148 m
9: Río Valdemembra (Júcar) en Villanueva de la Jara; 30SWJ96; 743 m
10: Río Bornova (Tajo) en San Andrés de Congosto; 30TVL93; 843 m
11: Río Sorbe (Tajo) en Valverde de los Arroyos, La Huerce; 30TVL85; 1060 m
12: Arroyo Alarconcillo (Guadiana) 200m aguas abajo del puente a Rochafrida, Ossa de
Montiel; 30SWJ10; 865 m
13: Río Vado Blanco (Guadiana) en Osero, Villahermosa; 30SWJ10; 880 m
14: Arroyo Fuente de la Parra (Segura) en Royo-Odrea, Ayna; 30SWH76; 851 m
15: Río Zumeta (Segura) en Las Juntas, Yeste; 30SWH42; 749 m
16: Río Montemayor (Júcar) en los Batanes, Peñascosa; 30SWH68; 1036 m
17: Río Fresnedas (Guadalquivir) en Cortijo de Cobatillas, Calzada de Calatrava; 30SVH26;
540 m
18: Arroyo Nava del Rey (Guadalquivir) en Huertezuelas, Calzada de Calatrava; 30SVH26;
591 m
19: Río Tietar (Tajo) aguas arriba del embalse Rosarito, Oropesa (Dehesa del Verdugal);
30TUK14; 317 m
20: Río Tiétar (Tajo) en Parrillas; 30TUK24; 371 m
21: Río Tiétar (Tajo) en Palacio Rosarito, Oropesa (Dehesa del Verdugal); 30TTK94; 289 m
22: Arroyo de las Ánimas (Guadiana) en Sotuélamos, El Bonillo; 30SWJ31; 882 m
23: Río Azuer (Guadiana) en Carrizosa, Montiel; 30SVH99; 811 m
24: Río Arroyo del cortijo (Bañuelos) (Guadiana) en Cortijo de Abajo, Los Cortijos;
30SVJ15; 739 m
25: Río Bullaque (Guadiana) en Casa de Durán, Piedrabuena; 30SUJ91; 539 m
26: Río Bullaque (Guadiana) en El Torno, Porzuna; 30SUJ94; 597 m
27: Río Estena (Guadiana) en Boquerón del Estena, Navas de Estena; 30SUJ67; 630 m
28: Río Córcoles (Guadiana) en Cortijo del Santo, Munera; 30SWJ41; 928 m
29: Río Gigüela (Guadiana) en Naharros; 30TWK43; 917 m
30: Río Gigüela (Guadiana) en Segóbriga, Saelices; 30SWK11; 778 m
31: Río Gigüela (Guadiana) en Villar del Horno; 30TWK53; 996 m
32: Río Guadiana (Guadiana) en Las Hoces-Casa Majalahoz, Puebla de Don Rodrigo;
30SUJ53; 446 m
33: Río Jabalón (Guadiana) en Montiel; 30SWH18; 869 m
34: Río Pinilla (Guadiana) en Villahermosa; 30SWJ10; 890 m
35: Río Riánsares (Guadiana) en Huelves; 30TWK03; 810 m
36: Río Riánsares (Guadiana) en Paredes; 30TWK13; 852 m
37: Río Tirteafuera (Guadiana) en Abenójar; 30SUJ70; 589 m
38: Río Valdeazoques (Guadiana) en Estación de Chillón, Chillón; 30SUH38; 376 m
39: Río Záncara (Guadiana) en Zafra de Záncara; 30SWK31; 899 m

35
40: Río Záncara (Guadiana) en Zafra de Záncara; 30SWK31; 829 m
41: Río Arquillo (Júcar) en Laguna del Arquillo, Masegoso; 30SWH58; 988 m
42: Río Arquillo (Júcar) en Peñascosa; 30SWH57; 1230 m
43: Río Cabriel (Júcar) en Balneario Fuente Podrida, Villatoya; 30SXJ45; 400 m
44: Río Cabriel (Júcar) en Cardenete; 30SXK10; 812 m
45: Río Guadazaón (Júcar) en Valdemoro de la Sierra; 30TXK04; 1121 m
46: Río Jardín (Júcar) en merendero El Zarzalejo, Casas de Lázaro; 30SWH69; 849 m
47: Río Júcar (Júcar) La Losilla, Alarcón; 30SWJ77; 729 m
48: Río Júcar (Júcar) en Albaladejito, Cuenca (Los Llecos); 30TWK63; 905 m
49: Río Júcar (Júcar) en la piscifactoría de Bolinches, Valdeganga; 30SXJ13; 602 m
50: Río Júcar (Júcar) en La Parra de las Vegas; 30SWK61; 824 m
51: Río Júcar (Júcar) en Villalgordo del Júcar; 30SWJ75; 670 m
52: Río Júcar (Júcar) en Casas del Palancar, Villar de Olalla; 30SWK62; 868 m
53: Río Júcar (Júcar) en Alarcón; 30SWJ77; 749 m
54: Río Júcar (Júcar) en Fuensanta; 30SWJ84; 659 m
55: Río Júcar (Júcar) en Valdemeca, Huélamo; 30TXK06; 1217 m
56: Río Júcar (Júcar) en Presa La Losa, La Losa; 30SWJ75; 680 m
57: Río Júcar (Júcar) en Molino de los Frailes, Valdeganga; 30SXJ03; 609 m
58: Río Júcar (Júcar) en Molino Juan Romero, Cuenca (Los Llecos); 30TWK95; 1178 m
59: Río Júcar (Júcar) en La Noguera (Barranco Boca de la Hoz), Tébar; 30SWJ87; 715 m
60: Río Júcar (Júcar) en Rambla Rubielos, Picazo; 30SWJ76; 695 m
61: Río Júcar (Júcar) en Villalba de la Sierra, Cuenca (Los Llecos); 30TWK74; 929 m
62: Río Júcar (Júcar) en Venta de Juan Romero, Huélamo; 30TWK95; 1180 m
63: Río Laguna (Júcar) en Huerta del Marquesado; 30TXK14; 1250 m
64: Río Masegoso (Júcar) en Masegoso; 30SWH58; 1130 m
65: Río Ojos de Moya (Júcar) en Garaballa; 30SXK30; 906 m
66: Río Pedregoso (Júcar) en Molino de Juan Romero, Beamud; 30TWK95; 1194 m
67: Río Valdemembra (Júcar) en Tarazona de la Mancha; 30SWJ94; 682 m
68: Río Vencherque (Júcar) en Villar del Humo; 30SXK11; 970 m
69: Arroyo Aliseda (Guadalquivir) en Carretera a Solana del Pino, Fuencaliente; 30SUH86;
701 m
70: Arroyo de la Aliseda (Guadalquivir) en Ventillas (Pueblo), Fuencaliente; 30SUH86; 684
m
71: Río Cereceda (Guadalquivir) en Cueva Batanera, Fuencaliente; 30SUH85; 822 m
72: Arroyo Cereceda (Guadalquivir) en Fuencaliente; 30SUH85; 663 m
73: Arroyo Dañador (Guadalquivir) en Cortijo de la Cañada, Villamanrique; 30SWH05; 761
m

36
74: Arroyo Caballeros del Escorial (Guadalquivir) en Casas del Escorial, Brazatortas;
30SUH76; 749 m
75: Río Guadalmena (Guadalquivir) en Villapalacios; 30SWH27; 754 m
76: Arroyo de las Navas (Guadalquivir) en Cortijo de las Navas, Solana del Pino; 30SUH95;
671 m
77: Arroyo Ontavia (Guadalquivir) en Hoz de Terrinches, Montiel; 30SWH16; 687 m
78: Río Pradillo (Guadalquivir) en Fuencaliente; 30SUH84; 552 m
79: Río Rigüelo (Guadalquivir) en San Lorenzo de Calatrava; 30SVH25; 524 m
80: Río Robledillo (Guadalquivir) en Solanilla del Tamaral, Mestanza; 30SVH15; 421 m
81: Río Robledillo (Guadalquivir) en Solanilla del Tamaral, Mestanza; 30SVH15; 459 m
82: Arroyo Ruicastaño (Guadalquivir) en Viso del Marqués; 30SVH46; 740 m
83: Río Villanueva (Guadalquivir) en Barranco Zárcenas, Villanueva de la Fuente;
30SWH27; 820 m
84: Río Yeguas (Guadalquivir) en Fuencaliente; 30SUH84; 564 m
85: Río Bogarra (Segura) en Batán de Bogarra, Bogarra; 30SWH67; 844 m
86: Arroyo Fuenfría (Segura) en El Encebrico (Chorraeros), Paterna del Madera; 30SWH56;
1431 m
87: Arroyo de las Hoyas (Segura) en El Puerto (Umbría), Paterna del Madera; 30SWH56;
1281 m
88: Arroyo Madera (Segura) en Arguellite, Yeste; 30SWH54; 706 m
89: Río Madera-Endrinales (Segura) en área recreativa Fuentelisa, Paterna del Madera;
30SWH57; 1044 m
90: Río Mundo (Segura) en el aforo del molino, aguas arriba de la piscifactoría, Riópar;
30SWH56; 937 m
91: Río Mundo (Segura) en Aguas abajo del embalse, Isso; 30SWH96; 470 m
92: Río Mundo (Segura) en Puente de Isso, Hellín; 30SXH05; 438 m
93: Río Mundo (Segura) en Liétor; 30SWH96; 515 m
94: Río Mundo (Segura) en Agramón; 30SXH15; 360 m
95: Río Mundo (Segura) en Mesones (aguas abajo del camping), Molinicos; 30SWH56; 886
m
96: Río Mundo (Segura) en Las Minas, Hellín; 30SXH14; 307 m
97: Río Mundo (Segura) en Tavizna, Hellín; 30SXH05; 402 m
98: Río Segura (Segura) en Calasparra (Est.de Las Minas), Hellín; 30SXH14; 301 m
99: Río Segura (Segura) en Cortijo de Abajo (aguas abajo del aforo), Hellín; 30SXH04; 349
m
100: Río Segura (Segura) en Salida Embalse Cenajo, Hellín; 30SXH04; 350 m
101: Río Segura (Segura) en El Hondo (Maeso), Hellín; 30SXH14; 334 m
102: Río Segura (Segura) en La Donal, Yeste; 30SWH53; 647 m
103: Río Segura (Segura) en Puente de Híjar, Férez; 30SWH95; 460 m

37
104: Río Segura (Segura) en Salmerón; 30SXH14; 310 m
105: Río Taibilla (Segura) en Nerpio; 30SWH52; 1092 m
106: Río Taibilla (Segura) en Molino la Tercia, Nerpio; 30SWH52; 1159 m
107: Arroyo Tobarra (Segura) en Saladar de Cordovilla, Hellín; 30SXH26; 500 m
108: Río Tus (Segura) en Vado de Tus, Yeste; 30SWH44; 835 m
109: Río Vadillos (Segura) en Yeguarizas, Bogarra; 30SWH66; 834 m
110: Río de la Vega (Segura) en El Laminador, Riópar; 30SWH56; 926 m
111: Arroyo Cedrón (Tajo) en Ablanque; 30TWL63; 1113 m
112: Río Ablanquejo (Tajo) en Ablanque; 30TWL62; 1008 m
113: Río Ablanquejo (Tajo) en Huertahernando; 30TWL52; 914 m
114: Río Alberche (Tajo) en Escalona; 30TUK74; 415 m
115: Río Alberche (Tajo) en Talavera de la Reina; 30SUK42; 367 m
116: Río Bornova (Tajo) en Molino de la Oportuna, Villares de Jadraque; 30TVL95; 971 m
117: Río Bornova (Tajo) en San Andrés de Congosto, La Toba; 30TVL93; 831 m
118: Río Bullones (Tajo) en Escalera; 30TWL81; 1032 m
119: Río Cañamares (Tajo) en Naharros, La Minosa; 30TWL05; 995 m
120: Río Cabrillas (Tajo) en Megina; 30TWK99; 1192 m
121: Arroyo Cedron (Tajo) en La Guardia, Corral de Almaguer; 30SVK70; 694 m
122: Río Cuervo (Tajo) en el nacimiento del Río Cuervo, Cuenca (Los Llecos); 30TWK97;
1428 m
123: Río Cuervo (Tajo) en Puente de Vadillos, Cañizares; 30TWK78; 943 m
124: Río Dulce (Tajo) en Jodra de Pinares; 30TWL34; 1079 m
125: Río Dulce (Tajo) en La Pelegrina, Sigüenza; 30TWL34; 980 m
126: Río Dulce (Tajo) en Villaseca de Henares, Villaseca de Henares; 30TWL13; 831 m
127: Río Escabas (Tajo) en el Estrecho de Priego, Priego; 30TWK67; 862 m
128: Río Escabas (Tajo) en Fuente del Cayo, Cuenca (Los Llecos); 30TWK77; 971 m
129: Río Gallo (Tajo) en Castilnuevo; 30TWL91; 1080 m
130: Río Gallo (Tajo) en Ermita de la Hoz, Corduente; 30TWL82; 1017 m
131: Río Guadarrama (Tajo) en Bargas, Bargas; 30SVK02; 452 m
132: Río Guadarrama (Tajo) en Chozas de Canales; 30TVK14; 515 m
133: Río Gévalo (Tajo) en Buenasbodas, Sevilleja de la Jara; 30SUJ39; 569 m
134: Río Guadiela (Tajo) en Canalejas del Arroyo; 30TWK47; 720 m
135: Río Henares (Tajo) en Azuqueca de Henares; 30TVK78; 604 m
136: Río Henares (Tajo) en Fontanar, Guadalajara (Monte el Villar); 30TVL80; 663 m
137: Río Henares (Tajo) en Jadraque; 30TWL03; 802 m
138: Río Henares (Tajo) en Mojares, Sigüenza; 30TWL35; 1056 m
139: Río Henares (Tajo) aguas abajo de la depuradora Sigüenza, Sigüenza; 30TWL24; 986 m

38
140: Río Henares (Tajo) en Tórtola de Henares; 30TVL80; 667 m
141: Río de la Hoz (Tajo) en Camping de la Venta (ruinas), Cantalojas; 30TVL76; 1329 m
142: Río Hoz Seca (Tajo) en Orea; 30TXK08; 1507 m
143: Río Jarama (Tajo) en El Cardoso de la Sierra; 30TVL54; 1240 m
144: Río Jarama (Tajo) en Casa de Uceda; 30TVL62; 744 m
145: Río Jarama (Tajo) en Dehesa Nueva del Rey, Seseña; 30TVK43; 484 m
146: Río Jarama (Tajo) en Puente Medieval, Valdesotos; 30TVL73; 803 m
147: Río Jarama (Tajo) en Puebla de Valles, Tortuero; 30TVL73; 802 m
148: Río Lillas (Tajo) en Barranco de Carretas, Cantalojas; 30TVL66; 1429 m
149: Río Lillas (Tajo) en parking del Parque Tejera Negra, Cantalojas; 30TVL76; 1404 m
150: Río Pusa (Tajo) en Bernuy, Malpica de Tajo; 30SUK61; 386 m
151: Río Sonsaz (Tajo) en área recreativa del Parque Tejera Negra, Cantalojas; 30TVL75;
1446 m
152: Río Sonsaz (Tajo) en Cabezo de la Mesta, Cantalojas; 30TVL75; 1657 m
153: Río Sonsaz (Tajo) en Sierra del Ocejón, Cantalojas; 30TVL75; 1541 m
154: Río Sorbe (Tajo) aguas arriba de la unión del Valdecimbrio, Galve de Sorbe; 30TVL86;
1254 m
155: Río Sorbe (Tajo) en Beleña del Sorbe, Cogolludo; 30TVL83; 803 m
156: Río Sorbe (Tajo) en Muriel, Tamajón; 30TVL83; 857 m
157: Río Tajuña (Tajo) en Abánades; 30TWL42; 1026 m
158: Río Tajuña (Tajo) en Archilla, Abánades; 30TWL00; 768 m
159: Río Tajuña (Tajo) en Loranca de Tajuna; 30TVK97; 700 m
160: Río Tajuña (Tajo) en Masegoso de Tajuna, Cifuentes; 30TWL21; 884 m
161: Río Tiétar (Tajo) en La Iglesuela; 30TUK55; 442 m
162: Río Tajo (Tajo) en Albáreal del Tajo, Polan; 30SUK91; 432 m
163: Río Tajo (Tajo) en Algarga, Illana; 30TVK94; 559 m
164: Río Tajo (Tajo) en Santuario Ntra Sra Carrasca, Villarrubia de Santiago; 30TVK63; 528
m
165: Río Tajo (Tajo) en Dehesa Nueva del Rey, Seseña; 30TVK43; 487 m
166: Río Tajo (Tajo) en Los Cuchillos, Peñalén; 30TWL80; 1099 m
167: Río Tajo (Tajo) en Malpica del Tajo; 30SUK61; 390 m
168: Río Tajo (Tajo) en Mocejon, Toledo; 30SVK21; 465 m
169: Río Tajo (Tajo) en Peralejos de las Truchas, Peralejos de las Truchas; 30TWK89; 1124
m
170: Río Tajo (Tajo) en Sacedón; 30TWK18; 690 m
171: Río Tajo (Tajo) en Las Herencias, Talavera de la Reina; 30SUK31; 357 m
172: Río Tajo (Tajo) en Trillo; 30TWL30; 749 m
173: Río Tajo (Tajo) en Valtablado del Rio; 30TWL50; 777 m

39
174: Río Tajo (Tajo) en Las Vegas y San Antonio, La Pueblanueva; 30SUK52; 377 m
175: Río Tajo (Tajo) en Zorita de los Canes; 30TWK06; 595 m
176: Río Trabaque (Tajo) en Arcos de la Sierra; 30TWK76; 980 m
177: Arroyo Valdecimbrio (Tajo) en su desembocadura en el Sorbe, Galve de Sorbe;
30TVL86; 1272 m
178: Río Zarzas (Tajo) en la entrada del Parque Tejera Negra, Cantalojas; 30TVL76; 1381 m
179: Río Zarzas (Tajo) en Barranco de las Lagunas, Cantalojas; 30TVL66; 1564 m

40
5.2. Descripción morfológica y ecología de algunas algas consideradas
‘raras’ en los ríos de Castilla – La Mancha.

Título original: Morphological description and ecology of some rare

macroalgae in south-central Spanish rivers (Castilla-La
Mancha Region)
Autores: José Luis Moreno, Laura Monteagudo y Marina Aboal
Publicación: Publicado en Anales del Jardín Botánico de Madrid, 70(1).
Impact Factor: 0.912
DOI: 10.3989/ajbm.2323
Fecha 2013

RESUMEN

El conocimiento sobre la biodiversidad algal de la región de Castilla-La Mancha,

situada en la zona centro-sur de España, es escaso en comparación con el de otras
regiones peninsulares. Sin embargo, la aplicación de la Directiva Marco del Agua
(2000/60/CE) y la evaluación del estado ecológico de los ecosistemas acuáticos
europeos, ha traído consigo un aumento en la frecuencia e intensidad en el muestreo
de ríos, lagos y humedales. De esta forma, durante los últimos años, se han
producido nuevos hallazgos en la región que han permitido ampliar el conocimiento
de la biodiversidad de algas así como de la distribución geográfica de muchas de sus
especies. En este trabajo se describen las condiciones ecológicas y las características
morfológicas de cinco especies que pueden considerarse raras a nivel europeo:
Nostochopsis lobata Wood ex Bornet et Flahault, Batrachospermum atrum (Hudson)
Harvey, Chroothece richteriana Hansgirg, Oocardium stratum Nägeli and
Tetrasporidium javanicum Möbius; y de una sexta especie, frecuente en España y
Europa, pero que supone la primera cita para esta región, Hydrurus foetidus (Villars)
Trevisan. Finalmente, se comparan las características morfológicas y ecológicas de
las poblaciones estudiadas con otras citas Europeas.

41
42
 Anales del Jardín Botánico de Madrid 70(1): 81-90, enero-junio 2013. ISSN: 0211-1322. doi: 10.3989/ajbm. 2323

Morphological description and ecology of some rare macroalgae

in south-central Spanish rivers (Castilla-La Mancha Region)
Jose Luis Moreno Alcaraz1*, Laura Monteagudo Canales1 & Marina Aboal Sanjurjo2
1
Centro Regional de Estudios del Agua, Universidad de Castilla-La Mancha, ctra. de Las Peñas km 3, E-02071 Albacete, Spain
2
Departamento de Biología Vegetal, Universidad de Murcia, Campus de Espinardo, E-30100 Murcia, Spain
joseluis.moreno@uclm.es; laura.monteagudo@uclm.es; maboal@um.es

Abstract
Moreno Alcaraz, J.L., Canales Monteagudo, L. & Aboal Sanjurjo, M. 2013.
Morphological description and ecology of some rare macroalgae in south-
central Spanish rivers (Castilla-La Mancha Region). Anales Jard. Bot.
Madrid 70(1): 81-90.
The Castilla-La Mancha Region (south-central Spain) is scarcely studied in
terms of freshwater algae. However, both the implementation of the Wa-
ter Framework Directive (2000/60/CE) and the evaluation of the ecological
state of European aquatic ecosystems have increased the intensity and fre-
quency of water body monitoring, including the rivers, lakes and wetlands
of this region. Thus, our knowledge on algal biodiversity and the geogra-
phical distribution of many species is rapidly increasing. In this study we
describe the occurrence, ecological conditions and morphological charac-
teristics of five algal species which are rare at the European level: Nosto-
chopsis lobata Wood ex Bornet & Flahault, Batrachospermum atrum (Hud-
son) Harvey, Chroothece richteriana Hansg., Oocardium stratum Nägeli
and Tetrasporidium javanicum Möbius. In addition, we include Hydrurus
foetidus (Vill.) Trev., a more common alga in Spain, since this is the first
record for the region. Finally, we compare morphological and ecological
characteristics of the studied populations with other European records.
Keywords: Algae, stream, river, Spain, Cyanophyceae, Rodophyceae,
Chlorophyceae.
Resumen
Moreno Alcaraz, J.L., Canales Monteagudo, L. & Aboal Sanjurjo, M. 2013.
Descripción morfológica y ecología de algunas macroalgas fluviales de la
España centromeridional (Castilla-La Mancha). Anales Jard. Bot. Madrid
70(1): 81-90 (en inglés).
El conocimiento sobre la biodiversidad algal de la región de Castilla-La
Mancha, situada en la zona centro-sur de España, es escaso en compara-
ción con el de otras regiones peninsulares. Sin embargo, la aplicación de la
Directiva Marco del Agua (2000/60/CE), y la evaluación del estado ecoló-
gico de los ecosistemas acuáticos europeos, ha traído consigo un aumen-
to en la frecuencia e intensidad en el muestreo de ríos, lagos y humedales.
De esta forma, durante los últimos años se han producido nuevos hallaz-
gos en la región que han permitido ampliar el conocimiento de la biodi-
versidad de algas así como de la distribución geográfica de muchas de sus
especies. En este trabajo se describen las condiciones ecológicas y las ca-
racterísticas morfológicas de cinco especies que pueden considerarse raras
a nivel europeo: Nostochopsis lobata Wood & Bornet & Flahault, Batra-
chospermum atrum (Hudson) Harvey, Chroothece richteriana Hansg., Oo-
cardium stratum Nägeli y Tetrasporidium javanicum Möbius; y de una sex-
ta especie, frecuente en España y Europa, pero que supone la primera cita
para esta región, Hydrurus foetidus (Vill.) Trev. Finalmente, se comparan
las características morfológicas y ecológicas de las poblaciones estudiadas
con otras citas Europeas.
Keywords: Algae, arroyo, río, España, Cyanophyceae, Rodophyceae,
Chlorophyceae.

INTRODUCTION “macrophytes and phytobenthos”. As a consequence, knowl-

The Castilla-La Mancha administrative region (south-central
Spain) remains as one of the most unknown of the Iberian Penin-
sula in relation to river algae diversity. Five main river basins
are included in this region: Tajo, Júcar, Guadiana, Guadalquivir
and Segura, but only the last has been intensely surveyed with
regards to algae (e.g. Aboal & Llimona, 1985; Aboal, 1988a-c,
1989a-c; Sabater & al., 1989; Aboal & al., 1996). The rest of re-
gion is scarcely studied (e.g. Aboal, 1996; Álvarez & al., 2007)
although a species list of Charophytes in Castilla-La Mancha
focusing mainly on wetland areas has been published by Ciru-
jano & Medina (2002). Additionally, a recent review on the sta-
tus of river aquatic plants in this region (Moreno & al., 2011)
provides an up to date regional catalogue of aquatic bryophyte
species as well as a list of macroalgae genera.
The implementation of the Water Framework Directive
(2000/60/CE) and the evaluation of the ecological state of Euro-
pean aquatic ecosystems have increased the intensity and fre-
quency of monitoring of water bodies, including the rivers,
lakes and wetlands of the study area. The assessment of the eco-
logical state of rivers by applying the Water Framework Direc-
tive (WFD) implies the monitoring of the biological element
edge on macroalgal biodiversity and the geographical distribu-
tion of many species has rapidly increased in recent years.
In this study we describe the occurrence, ecological condi-
tions and morphological characteristics of some uncommon al-
gal taxa. One of them has been the first record for Spain and the
third for Europe: Nostochopsis lobata Wood ex Bornet & Fla-
hault; four additional species are hardly cited in Europe: Batra-
chospermum atrum (Hudson) Harvey, Chroothece richteriana
Hansg., Oocardium stratum Nägeli and Tetrasporidium java-
nicum Möbius; and finally, Hydrurus foetidus (Vill.) Trev.,
which has been collected in cold streams of some mountains of
Spain although our record is the first cite for the study area.
All the taxa were found within the boundaries of the Au-
tonomous Community of Castilla-La Mancha (south-central
Spain) (Fig. 1) which occupies an area of 79409 km2. This re-
gion includes the upper and middle reaches of five large river
basins: Tajo, Guadiana, Guadalquivir, Júcar and Segura. Land
uses are mainly agriculture (46 % of the regional area) and fo-
rest (44 %). Regarding geology, three zones can be distin-
guished: the western zone is rich in Precambrian siliceous
rocks (mostly quartzite, slate, shale, granite and gneiss); Meso-

* Corresponding author.
43
82 J.L. Moreno Alcaraz & al.
Fig. 1. Map of the Iberian Peninsula showing the limits of the Castilla-La Mancha
Region, the main rivers crossing the study area and the location of the species
recorded. ♦ Hydrurus foetidus; ☐ Oocardium stratum; ● Nostochopsis lobata; ▲
Chroothece richteriana; ◯ Batrachospermum atrum; ■ Tetrasporidium javanicum;
Castilla-La Mancha Region.
zoic calcareous rocks (limestone, dolomite, sandstone and con-
glomerates) are dominant in the eastern area; and finally, Ter-
tiary sedimentary fills are accumulated in the great central
plateau located at 700 m a.s.l. called “La Mancha”, where
clays, sandstones, gravels, stones, conglomerates, marls and
gypsum are predominant (González & Vázquez, 2000). Moun-
tains are located mainly on the edges of the region and can
reach more than 2000 m (2273 m, Pico del Lobo)
MATERIAL AND METHODS
As part of a regional river monitoring program, a stream
reach of approximately 100 m long was deeply surveyed for
macroalgae, including all microhabitats present, e.g. riffles,
pools, runs or stream margins as well as different kinds of sub-
strata (sand, gravel, stones, aquatic vegetation, etc.). Macroal-
gal samples were collected in the river by hand and taken to the
laboratory. One part of the samples was fixed in 3 % formalde-
hyde in the field whereas the other part was maintained fresh
until laboratory observations. Collected material was exam-
ined under a Leica M165C stereoscope and a light microscope
OLYMPUS BX50. Glycerin-gelatine was used to make per-
manent slides which were used to take cell measurements.
Drawings were made with the help of macroscopic and micro-
scopic images taken by a Leica DFC 420 C camera and also by
direct observation of samples. Electric conductivity, pH and
dissolved oxygen were measured in situ using appropriate sen-
sors (Multiline P4 WTW). Alkalinity was also obtained in the
field using the sulphuric acid method (APHA, 1998). In addi-
tion, water samples were collected in polyethylene bottles (500
ml) and were kept in the refrigerator at 4 ºC. All physico-chem-
ical parameters were analyzed within 48 h after sampling. The
44
concentration of nutrients (N-NO3-, N-NO2-, N-NH4+, and P-
PO4-3) was determined photometrically with MERCK Kits
(Spectroquant®); ion chromatography was used to analyse
chloride and sulphate; turbidity with a turbidimeter TN-100
(Eutech Instruments; infrared light); calcium and magnesium
by complexometry (volumetry); sodium and potassium by
atomic emission. All these parameters were analysed following
standard procedures detailed in APHA (1998).
RESULTS
Nostochopsis lobata Wood ex Bornet & Flahault (Fig. 2a-c)
Some specimens of N. lobata were found at the Nava del Rey
stream (Fig.1), a temperate temporary stream tributary of the
Guadalquivir river. The stream was located in the southern part
of Castilla-La Mancha (Province of Ciudad Real), and it flows
over Palaeozoic siliceous metamorphic rocks (quartzites, slates
and shales). The study site was located at medium altitude (590
m) at 9.5 km from the source with an upstream drainage area of
41.76 km2. N. lobata was found when the stream was at base
flow condition (June 2009). Two morphological forms were
collected: compact globular specimens corresponding to young
colonies (Fig. 2a,b) were found attached to a moss stem where-
as free floating diffluent fragments coming from senescent
colonies were collected entangled with other macrophytes.
Young colonies measured 0.5-3 cm in diameter while diffluent
fragments were about 0.1-1 cm; and its colour changed from
brownish to bluish, respectively. The radial disposition of fila-
ments could be clearly observed in the outer part of the
colonies, in transversal sections (Fig. 2b). Cells were isodia-
metrical to cylindrical, measuring from 2.0 µm up to 6.17 µm in
diameter and length up to 7 µm. Heterocytes were predomi-
nantly lateral, sessile or pedicellate (Fig. 2c), measuring from
6.7 µm to 8.0 µm in diameter. Intercalary heterocytes were ra-
re. The waters were oligo-mesotrophic, with low conductivity
and alkalinity values (Table 1). The specimens were collected
at marginal depositional habitats along with other aquatic
macroflora such as Ranunculus peltatus Schrank, Cladophora
sp., Spirogyra sp., Oedogonium sp., Zygnema sp. and Tetras-
poridium sp.
Batrachospermum atrum (Hudson) Harvey (Fig. 2d-f)
B. atrum was found at three study sites: the Vado Blanco
stream, Cabriel stream and Júcar river (Fig. 1). The Vado Blan-
co stream connects two lagoons of the Ruidera lagoon complex
made up of 15 karstic lagoons located on a high altitude cal-
careous plateau (around 900 m a.s.l.) where forest and irriga-
tion land are the main land uses. Nitrate concentration in
groundwater and surface water is high due to intensive agricul-
ture practises developed in the last decades (Berzas & al.,
2004). The Cabriel Stream is the main tributary of the Júcar
River and the study site was located at a high altitude near its
source (1156 m a.s.l.) in a mountainous area where forest is
predominant. Groundwater inputs from springs and upwellings
are important in this stream reach. The last study site was lo-
cated in a middle reach of the River Júcar at medium altitude
(600 m) in a wide valley dedicated to agricultural uses. The in-
fluence of a reservoir located 96.65 km upstream is very signi-
 Rare macroalgae in south-central Spanish rivers 83

ficant due to flow regulation for agriculture. The river reach is

(Vill.) Trev.

26/03/2012
Hydrurus
foetidus

<0.010
siliceous

0.02

7.53

7.48
16.61
0.92
0.41
0.98
Zarzas
also influenced by groundwater upwelling and seepages that

1564

2.4

7.8

9.5

0.5

0.1
12
0

8
contribute to feed the river. The specimens were collected sub-
merged in the stream margins, mainly over other macroalgae or
entangled with them and exposed to a low current velocity.
Thallus size ranged from 2.5-8 cm long and 60-120 µm wide,
Nava del Rey
28/04/2010

siliceous

colour red-brownish, without mucilage (Fig. 2d and 2e), show-

9.11

<0.03
0.33

104.00
15.44
41.50
23.00
10.00
8.57
1.27
n.d.
590

3.5

24.5
8.3
ing a regular cortication (Fig. 2f) along its central axis and
Tetrasporidium

70

193
javanicum

branches. The mean distance between nodes was of 250-525

Möbius

µm and decreasing from the central axis base towards the api-
cal branches (up to 80 µm in branch tips). The whorls were very
13/05/2002

0.112
siliceous

reduced, 100-200 µm diameter, non-adherent to the main axis,

Estena

0.08

7.46
7.85

0.08

12.82
8.93

2.24
3.73
816

1.5

13.7

0.9

15.5

1.1
and ring or pearl shaped (obconical). Primary fascicles were
29

30 composed of 3-6 barrel-shaped cells of 3.3-6.8 µm in diameter.

Secondary fascicles were composed of 1-3 cells, abundant and
scattered along the internodes or concentrated in nodes, the api-
Fte. de la Parra

cal cell of fascicles usually sharp-pointed. Gonimoblasts were

Oocardium

27/02/2012

calcareous

0.001

0.027
stratum

hemispherical, 135-220 µm in diameter and 27-80 µm high, at-

Nägeli

7.91
9.21

19.47
31.05
55.17
25.19
10.73
0.91
851

11.5

2.4

tached to the axis in internodes or nodes. Tricogyns shape was

1
–

547

284

clavate, 9.3-16.2 µm long and 6.2-8.3 µm wide, located main-

ly in internodes but also in nodes and axillas at the base of
branches. Tricogyns usually included 1-2 round spermatia of
4.7-6 µm diameter attached to the tip. Regarding environmen-
Chroothece
richteriana

29/05/2007

calcareous

0.137
Bullones

tal conditions, it is important to highlight that the three locali-

Hansg.

7.34
9.81

1.14

383.02
313.66
126.12

174.76
4.62
1032

12.9

53.1

ties were under groundwater influence. Dissolved nutrients

2
–

2470

0
288

showed low values except for one site (Vado Blanco stream,
Lagunas del Ruidera) located at an irrigation area with high
levels of nitrate contents (Table 1). Accompanying aquatic
03/11/2009

calcareous

0.746
0.019

macroflora included Anabaena sp., Batrachospermum gelati-

0.13

23.68
167.65

25.39
13.19
2.18
Júcar

607

9.8
1.8
15.6
8.1
9.2

161.3

77.2

nosum (L.), Chara aspera C.L. Dethard. ex Willd. and Chara

847

hispida L. in Vado Blanco stream; Apium nodiflorum (L.) Lag.,

B. gelatinosum and Bryum sp. in Júcar river; and Apium nodi-
Batrachospermum

florum, Audouinella sp. and B. gelatinosum in Cabriel stream.

atrum (Hudson)

30/03/2005

calcareous

0.029

0.007
Cabriel

0.38

7.98

0.44

6.71
85.52

2.85
0.64
Harvey

1156

6.9

13.5

15.5

Chroothece richteriana Hansg (Fig. 3a-c)

577

212

67
41

C. richteriana was found in Bullones stream (Fig. 1), a head-

water tributary of the River Tajo situated in a calcareous land-
scape in contact with marls and limestones. The drainage area
Vado Blanco
27/05/2009

of subcatchment was 137.97 km2, with 30 % of the area occu-

calcareous

0.185

0.045
11.103
0.02

48.45
114.19
110.71
19.35
24.17
1.79
860

2.5

19.9

8.8

218.1

pied by agriculture and 70 % by natural vegetation. The imme-

Table 1. Geographical and ecological data for the algae species studied.

870

diate vicinity of the site was a semi-natural area mainly occu-

pied by Juniperus communis L. and J. thurifera L. The site was
located at 1032 m a.s.l., 25 km downstream from the source. In
general, colonies of this species were bright green, gelatinous
Nostochopsis lobata

and hemispheric in shape. Transversal sections revealed a strat-

Wood ex. Bornet

ified structure of the hemispherical colony (Fig. 3a): the sur-

Nava del Rey
09/06/2009
& Flahault

0.042

0.001
siliceous

0.03

7.47
6.84

0.33

15.44

8.57
1.27

face was conformed by a continuous layer of cells on the top of

590

3.5

18.8

41.5
237

92

23
10

their radially-arranged stalks, with some parallel lines of sedi-

ments and carbonate precipitates probably corresponding to
growth rings. C. richteriana was found growing epilithic, cov-
ering around 80 % of most submerged rocks. Cells were cylin-
conductivity 25º (μS·cm-1)
dissolved oxygen (mg·L-1)
mean channel width (m)
lithology (predominant)

ortophosphate (mg L-1)

drical and elongated, measuring from 5 µm to 9 µm in width

magnessium (mg L-1)
ammonium (mg L-1)

(6.9 µm average) and from 10 µm to 18 µm in length (15.9 µm

date (dd/mm/yyyy)

potassium (mg L-1)

alcalinity (mg L-1)
discharge (m3·s-1)

sulphate (mg L-1)

chloride (mg L-1)

calcium (mg L-1)

average) (Fig. 3b,c). The cells were surrounded by a thick cell

sodium (mg L-1)
nitrate (mg L-1)
River/Stream

wall (from 2 to 6 µm wide) and disposed on the stalks (Fig. 3b).

altitude (m)

Inside some cells, a star-shaped chromatophore was visible

Species

T (ºC)

(Fig. 3c). Regarding physico-chemical conditions (Table 1)

pH

water was slightly saline due to the presence of marls alterna-

45
84 J.L. Moreno Alcaraz & al.
Fig. 2. Nostochopsis lobata: a, macroscopic view; b, transversal section; c, mature filament with heterocytes (from Moreno & al., in press). Batrachos-
permum atrum: d, macroscopic view; e, filament; f, detail of gonimoblast and cortication of the thallus. Scale: a, d = 50 mm; b = 1 mm; c, f = 50 μm;
e = 500 μm.
ting with limestones (I.T.G.E., 2000), with a high content of
chloride and sulphate. An industry of salt extraction is located
8 km upstream (Salinas de Almallá) due to the presence of
evaporite outcorps.
Other aquatic taxa detected at the site were: Apium nodiflo-
rum, Veronica anagallis-aquatica L., Chara vulgaris sp.,
Cladophora sp., Tetraspora sp., Vaucheria sp., Batrachosper-
mum sp., Tolypothrix sp., and Phormidium sp.
Oocardium stratum Nägeli (Fig. 3d-i)
The desmid zygnematal O. stratum was found in Fuente de
la Parra stream (Fig. 1), a small tributary of the Mundo River
(Segura river basin). This stream runs over calcareous rocks in
a mountainous area covered by a Pinus pinaster Ait. forest. The
site was located at 851 m a.s.l. very close to the source (1.73 km
downstream). The biodiversity and proliferation of macroalgae
was low, due to the dense canopy reducing the entry of light.
O. stratum appeared on mosses, sticks and wet rocks in a small
dam 3 m in height built to retain water from the source, where
a travertine formation was extensive across the waterfall. To
the naked eye, colonies first formed hemispherical structures
like small grains a few millimetres long (Fig. 3d). However,
with the growth and the increase of carbonate precipitates, the
colonies formed a lobulated layer of calcareous concretions
46
several centimetres thick on twigs and sticks (Fig. 3e). Under
the microscope, a lateral vision of a colony (Fig. 3b) showed
that this species formed gelatinous branched cylinders sur-
rounded by calcite with bright green cells at the top (Fig. 3g).
Cells were heart-shaped, showing a small median constriction
as in other desmids such as Cosmarium, and measured around
13.5 µm in diameter (11-16 µm) (Fig. 3h).
In some cases, colonies of Chroothece rupestris Hangs.
were found over colonies of O. stratum. After using acetic acid
to eliminate carbonate precipitates from Oocardium colonies,
filaments of Rivularia sp. were observed inside them (Fig. 3i).
These three algae species, together with the moss Didymodon
tophaceus (Brid.) Lisa, were revealed to be the main biological
builders of the global travertine structure. The accompanying
aquatic macroflora included Callitriche stagnalis Scop., Vero-
nica anagallis-aquatica, Chara vulgaris, Batrachospermum
gelatinosum, Phormidium sp., Oedogonium sp., Cladophora
glomerata (L.) and Microspora sp.
Tetrasporidium javanicum Möbius (Fig. 4a-e)
In the study area, this clorophyte was found at two sites. The
first site was Estena stream (Fig. 1), a small tributary of the
Guadiana river running over siliceous rocks. The study site was
located in a mountainous area of the Cabañeros National Park,
 Rare macroalgae in south-central Spanish rivers 85

Fig. 3. Chroothece richteriana: a, transversal section of a colony; b, lateral view of pedunculated cells; c, cells with star-shaped chromatophores. Oocar-
dium stratum: d, small colonies covering a moss branch; e, aged colony forming a thick lobulated layer; f, lateral microscopic view showing the calcite
cylinders surrounding cell stalks; g, upper view of cells surrounded by calcite; h, detail of cell with stalk; i, colony of O. stratum together with Rivularia af-
ter carbonate removal. Scale: a, i = 500 μm; b, c, h = 20 μm; d = 5 mm; e = 1cm; f = 100 μm; g = 200 μm.

and 6.1 km downstream from the source. The drainage area
was covered by a near natural forest of Quercus Mediterranan
maquia. T. javanicum appeared as small (environ 8 mm in
length) free floating colonies forming a light green gelatinous
tubular-net thallus (Fig. 4a). These small colonies probably
corresponded to fragments of a bigger colony. The ‘ribbons’
making up the thallus (Fig. 4b) measured 100-300 µm wide and
presented revoluted margins. Cells were heterogeneously ar-
ranged in the mucilage and were spherical to slightly oval in
shape (Fig. 4c). A wide range of cell size was observed: from
5.1-10.7 µm in diameter, with 8.2 µm on average. Chloroplas-
ts were parietal with a single large pyrenoid (around 1.6 µm in
diameter). Pseudo-flagella, typical of the genus Tetraspora,
were absent. Environmental and physico-chemical data record-
ed for this site indicated that the stream was mesotrophic,
showing a very low mineralisation due to the siliceous litho-
logy (Table 1). The waters showed a carbonated character but
with a high proportion of chloride. Accompanying aquatic
macroflora included Audouinella sp., Draparnaldia sp., Zyg-
nema sp., Tolypothrix sp., Scytonema sp., Nostoc sp., and Chi-
loscyphus polyantos (L.) Corda.
In addition, we collected Tetrasporidium in another
siliceous oligo-mesotrophic stream, the Nava del Rey stream
47
(described in Nostochopsis lobata section), but different mor-
phological characteristics were observed. In this case, colonies
were thin perforated sheets (Fig. 4d), the biggest one measur-
ing 1.5 ⫻ 2.1 cm. Cells were smaller and surrounded by con-
centric gelatinous sheaths (Fig. 4e). The smaller cell size (from
3.3 µm to 5.7 µm in diameter) of this specimen matched with
the description of T. fottii Couté et Traccana included in the key
of Ettl & Gärtner (2009), although for other authors Tetras-
poridium is a monospecific genus (see discussion).
Hydrurus foetidus (Vill.) Trev. (Fig. 4f-h)
H. foetidus was found at Zarzas stream (Fig. 1), a headwater
tributary of the Hoz stream which belongs to the largest Span-
ish river basin, the Tajo river. The Zarzas stream runs through
the Natural Park of Sierra Norte of Guadalajara, located in the
northernmost part of the region. This area is mountainous and
is covered by a forest of Pinus sylvestris L. with the presence of
Fagus sylvatica L. The study site was located at an altitude of
1564 m, in one of the coldest areas of the region, 7.90 km
downstream from its source. The drainage area of subcatch-
ment is 26.90 km2. This alga was found as gold-brown tufts up
to 20 cm long attached to boulders and stones (Fig. 4f,g) some-
86 J.L. Moreno Alcaraz & al.

Fig. 4. Tetrasporidium javanicum: a, macroscopic view of a tubular-net colony (“ribbon-like”); b, detail of a part of the colony; c, detail of cells; d, macro-
scopic view of a laminar colony; e, detail of the colony showing cells surrounded by gelatinous sheaths. Hydrurus foetidus: f,g, macroscopic view of two
thallus shapes; h, detail of cell arrangement within the mucilage. Scales: a = 2 mm; b = 100 μm; c, h = 20 μm; d = 500 μm; e = 50 μm; f, g = 1 mm.

times covering 100 % of the substratum. Thalli were branched
and gelatinous. Cells embedded in the mucilage were peripher-
ally arranged and oval to ellipsoidal in shape (Fig. 4h). Cell size
was 5-8 µm wide (6.5 µm on average) and 7-13.5 µm long
(10.5 µm on average) in. Regarding nutrients, waters were oli-
go-mesotrophic (Table 1). Accompanying aquatic macroflora
was made up of the following taxa: Lemanea sp., Hildenbran-
dria sp., Scapania undulata (Hedw.) P. Beauv., and Marchan-
tia polymorpha L.
DISCUSSION
N. lobata is included in the family Nostochopsaceae (Anag-
nostidis & Komarek, 1990; Komárek & al., 2003). This species
is characterized by a globular lobed thallus composed of unise-
riate and branched thricomes with intercalar or lateral hetero-
cytes (pedicellate or sessile) (Bornet & Flahault, 1886-1888).
N. lobata is widely distributed in tropical areas of the world and
has been reported in several African countries (Frémy, 1929),
North and South America and Australia (Cáceres, 1973; Sarma
& Chapman, 1975; Branco & al., 2001) and Asia (Geitler,
48
1930-1932; Desikachary, 1959; Tiwari, 1978; Peerapornpisal
& al., 2006). In Europe there are only third records from flow-
ing warm waters: Banyuls-sur-Mer (France) (Frémy & Feld-
mann, 1934), Corsica (France) (Hoffmann, 1990), and L’Aqui-
la (Italy) (Del Grosso, 1977). The specimens of N. lobata col-
lected in Nava del Rey were the first recorded in Spain (Mo-
reno & al., in press). Morphologically, our material matched
fairly well the detailed description of the first specimen report-
ed in Europe by Frémy & Feldmann (1934). In both cases N. lo-
bata was found in small temporary streams running over
siliceous metamorphic rocks, located at low altitude and living
in warm and oligotrophic waters, and it was collected in sum-
mer. In terms of habitat, we found N. lobata growing epiphytic
(young) and free floating (senescent) even though it has been
reported in different habitats: lakes (Tiwari, 1978; El Saied,
2007), running waters (Frémy & Feldmann, 1934; Palmer,
1941; Pandey & Pandey, 2008), thermal springs (Yoneda,
1939), epiphytic (Sarma & Chapman, 1975), attached to rocks
(Hoffmann, 1990; Peerapornpisal, 2006), free floating (Wood,
1872), on moist soils (Skinner & Entwisle, 2001; Aziz, 2008),
and on humid walls (Skinner & Entwisle, 2001) among others.
 Rare macroalgae in south-central Spanish rivers
However, the most extreme habitat where N. lobata has been
found is probably as cryptoendolithic in an arid climate (Weber
& al., 1996). Current data point out that this species is dis-
tributed worldwide with an underlying tendency: it grows
abundantly in tropical regions where its optimal climatic con-
ditions are stable for several months whereas it grows scarcely
in other regions only when conditions allowing its germination
and growth are reached, probably for a few weeks in occasion-
al years (Moreno & al., in press). This fact explains the diffi-
culty in detecting this species in Europe, where it has always
been recorded only once per site.
B. atrum belongs to subgenus Batrachospermum, which is
characterized by a carposporophyte multicellular, and to sec-
tion Setacea which is characterized by short carpogonium-
bearing branches and whorls/fascicules (Eloranta & Kwan-
drans, 2007). B. atrum (section Setacea) is on divergent
branches of molecular trees (Kumano, 2002) and constitutes a
distinct section, as proposed by Sheath & al. (1993). Goni-
moblasts form mamilliform or semispherical swellings on the
central axis and the trichogyn is clavate; the thallus is subge-
latinous (Starmach, 1977). The habitus shows a compressed
and compact thallus due to its short fascicules and whorls, re-
sembling a bamboo stem, similar to the thallus of the red algae
Lemanea. According to Kumano (2002), B. atrum is distribut-
ed throughout Europe (UK, France, Belgium, Germany,
Poland, Portugal and Sweden), Eastern Asia (China, Korea and
Japan), Australia, New Zealand, South America (Brazil) and
Africa (Angola), in addition to the USA (California and Texas;
Sheath & al, 1993). In Spain there is only one previous record
from the Murcia Region (Aboal & al., 1995). The morphologi-
cal characteristics match with other descriptions provided by
other authors (Israelson, 1942; Sheath & al., 1993; Eloranta &
Kwandrans, 2007). B. atrum is an oligotrophic hard water
species (Sládecek, 1973; Rott & al., 1999; Eloranta & Kwan-
drans, 1996, 2002). In Spain, Aboal & al. (1995) found this
species in calcareous springs and small streams with environ-
mental conditions similar to those described in this work. The
three sites where B. atrum was collected were calcareous and
they showed groundwater influence associated to upwelling ar-
eas, springs or seepages. The trophic conditions in our sites
were oligo-mesotrophic, with one site showing a high concen-
tration of dissolved nitrate due to its location within an inten-
sively irrigated cropland area (Álvarez-Cobelas & al., 2005).
According to Eloranta & Kwandrans (2007) the species grows
epiphytic or epilithic, but in our sites it was collected as epi-
phytic, floating among other macrophytes close to river banks
and avoiding high current velocity, more limnephilic than
rheophilic. According to Israelson (1942) and Aboal & al.
(1995) the species seem to occur only in spring, but in the study
area the species was recorded both in spring and autumn, simi-
lar to the seasonal pattern frequently shown by the rest of the
Batrachospermaceae in lotic waters (Aboal & al., 1995). The
fact that the species was collected only once per site despite
several visits may be due to the rapid development and short life
cycle of the species or to the occasional interannual occurrence.
After C. richteriana was described by Hansgirg (1884), few
publications about this species have been released. C. richte-
riana has been reported in Britain (Sheath & Sherwood, 2002;
Eloranta & al., 2011) and Spain (Eloranta & al., 2011). Recent-
49
87
ly it has been found growing in slightly saline streams of south-
eastern Spain (Murcia), mostly on subaerial conditions (un-
published data). This rare Rhodophyte included in the Por-
phyridiaceae family is characterized by mucilaginous colonies
conformed by elliptical cells enclosed in a layered gelatinous
matrix (Eloranta & Kwandrans, 2007; Eloranta & al., 2011).
Additionally, a star-shaped chromatophore is clearly visible in-
side the cells. Starmach (1977) and Eloranta & al. (2011) con-
sider cell size as a differential characteristic between the two
species of Chroothece present in Castilla-La Mancha: C. rich-
teriana measuring 6-10 µm wide and 15-18 µm long; C. ru-
pestris measuring 5-7 µm wide and 9-15 µm long. Additional-
ly, Sheath & Sherwood (2002) reported a cell size for C. rich-
teriana of 8-12 µm wide and 10-21 µm long (British Islands).
Both studies match with the cell size of our colonies collected
in Bullones stream (5-9 µm wide, 10-18 µm long). However,
we noticed that according to our observations, the cell size
range of a Chroothece population can exceed that one reported
from identification keys. On the other hand, the relatively high
water salinity of Bullones stream was in accordance with the
“more or less saline” environment described for C. richtheria-
na in the identification keys (Eloranta & al., 2011; Starmach,
1977).
Regarding Chroothece populations from Fuente de la Parra
(see Oocardium stratum section), they were identified as C. ru-
pestris according to the identification keys criteria: lower cell
size, low water mineralisation, and also the environment “wet
rocks”. In the same way, Margalef (1989) recorded C. rupes-
tris from wet rocks of calcareous springs, described by him as
“hygropetric environment”, a similar environment to that of
Fuente de la Parra stream.
Oocardium is a monospecific genus belonging to the family
Desmidiaceae, characterized by being mostly unicellular, ei-
ther solitary or in colonies, with cells divided into two com-
partments separated by an isthmus. West (1904) described
Oocardium as the most extraordinary of all the genera of
Desmids usually occurring in large colonies. According to
West (1904) and Bourrelly (1990), the colonies of O. stratum
are hemispherical in shape (1-2 mm in diameter), and are
formed by cells standing on more or less parallel, radiating
strands of mucus which are encrusted in calcite. During the
growing period, calcite crystals are formed by continuous de-
position by surrounding cells which, following division, calci-
fy into slightly different directions upwards (Sanders & Rott,
2009). The colonies detected in Fuente de la Parra stream
matched exactly with the above description and appeared joint-
ed, forming a nodular crust covering substrates, similarly to
colonies observed by Pentecost (1991). Regarding the environ-
mental conditions, O. stratum seems to have very specific re-
quirements: it is only found in waterfalls and springs with cal-
careous water (Margalef, 1983; Pentecost, 1991) usually being
associated to deposits of travertine and tufa (Pentecost, 2001;
Sanders & Rott, 2009; Bellinger & Sigee, 2010). Because of
these environmental restrictions, O. stratum is often considered
as an unusual alga (e.g. Margalef, 1983; Bellinger & Sigee,
2010) even though its presence has been detected worldwide:
Switzerland (Nägeli, 1849) the United States of America
(Prescott, 1981), Austria (Sanders & Rott, 2009), United King-
dom (West, 1904; Pentecost, 1991), Ireland (Carter, 1923),
88 J.L. Moreno Alcaraz & al.
Belgium (van Oye & Hubert, 1937), France (Kouwets, 1999),
Slovenia (Vrhovsek & al., 2006). Particularly in Spain, O. stra-
tum has been previously reported in Barcelona (Margalef,
1955), Murcia (Aboal, 1991) and in Castilla-La Mancha
(Ruidera Lakes) (Álvarez-Cobelas & al., 2007), although we
think that it could be a common species in calcareous moun-
tains and springs.
T. javanicum is a clorophyte classified within the order
Tetrasporales, family Palmellopsidaceae (Ettl & Gärtner,
2009). In the mature state the thallus is a thin, flat, more or less
circular sheet with numerous irregular perforations with a
smooth or lobed margin (Iyengar, 1932). Nevertheless, the
specimen collected in the Estena River was a tubular-net thal-
lus, probably as a result of the increase in the size perforation
with age. This fact supports the idea that ribbon-like and sheet-
like thallus could represent plants of different ages (Entwisle &
Skinner, 2001). According to Ettl & Gärtner (2009) cells are
oval to spherical in shape ranging from 6 µm to 15 µm in diam-
eter, slightly larger than our material (from 5.1 to 10.7). Re-
garding distribution, this species is abundant in tropical areas
within India (Iyengar, 1932), China (Hu & Wei, 2006), Hawai-
ian Islands (Sherwood, 2004), Java, the United States, Australia
and Bangladesh (Entwisle & Skinner, 2001) being quite com-
mon in tropical areas (Entwistle & Skinner, 2000). In Europe,
T. javanicum has been reported in the Czech Republic, France,
Portugal and Spain (Fott & al., 1965; Coute & Tracanna, 1981;
Calado & Rino, 1992; Aboal & al., 1994, respectively). In
Spain, this species has been found in the Alicante Province
(Aboal & al., 1994), Cáceres Province (Marín-Murcia & Aboal,
2007), Galicia (López-Rodríguez & Penalta-Rodríguez, 2007)
and, recently, in the Ebro river basin (NE Spain) (Tomás & al.,
in press). In almost all cases, the species is found growing in
conditions of low current velocity, well oxygenated alkaline
waters, high turbidity and nutrient enrichment due to agricul-
tural practises. However, the material described in this work
was found in low-mountain clear water streams. These condi-
tions are more similar to those described by Calado & Rino
(1992) and López-Rodríguez & Penalta-Rodríguez (2007) for
specimens collected in Portugal and Galicia, respectively. De-
spite the fact that the Estena stream is oligotrophic, T. javani-
cum was found coinciding with a relatively high concentration
of nitrogen compounds (nitrite, nitrate and ammonium). This
fact could be due to the preference of T. javanicum for eutroph-
ic waters, as suggested by some authors (Aboal & al., 1994;
Marín-Murcia & Aboal, 2007; Tomás & al., in press) pointing
to this species as a potential good indicator of eutrophy. Our ob-
servations lead us to think that T. javanicum is more common
than reported up to now in Spain and it will be collected more
frequently in upcoming river macrophyte surveys.
Regarding T. fottii, the species was described by Couté &
Tracanna (1981) from some specimens found in Breton ponds.
However, we were able to find only one other report on this
species in literature, made by Neag & al. (2005) in the "Alexan-
dru Borza" Botanical Garden, Cluj-Napoca, Romania. Ettl &
Gärtner (2009) differenced T. fottii from other Tetrasporidium
species according to the lateral position of the pyrenoid and its
cell size (from 6.4 µm to 9.5 µm in diameter). However, the
material collected in Nava del Rey was slightly smaller, mea-
suring from 3.3 µm to 5.7 µm in diameter. Unfortunately, an
50
appropriate observation of the ultrastructure of cells was not
possible and, like Entwisle & Skinner (2001), we could not
conclusively identify our material as T. fottii. On the other
hand, Calado & Rino (1992) suggest that the specific charac-
teristics of T. fottii could be explained by the cytologic vari-
ability within Tetrasporidium javanicum and, therefore, T. fot-
tii should not be considered as an independent species. This
idea is contemplated by other authors that classify Tetraspori-
dium as a monospecific genus (e.g. Bourrelly, 1990). Thus, we
assigned our material to T. javanicum until more studies unra-
veling the taxonomy of this interesting genus are released.
The chrysophyte Hydrurus foetidus is included in the order
Chromulinales, family Hydruraceae. The thallus is gelatinous,
branched and attached to substrate reaching up to 30 cm long.
Cells are ellipsoid to subspherical and they are randomly ar-
ranged in the mucilage (Rodriguez & Vergon, 1996). After col-
lecting, it emanates a characteristic smell. This species is a cold
water stenotherm also adapted to strong current velocity, being
an exclusive inhabitant of cold mountain streams worldwide
(2-12 ºC) (Hieber & al., 2001; Wehr & Sheath, 2003; Kriz-
manic & al., 2008). Under optimal conditions, H. foetidus be-
comes dominant and may entirely cover submerged rocks,
making them very slippery. In Europe it is a well-known inhab-
itant of alpine streams and those draining glaciated landscapes,
where it is described as a predominant or very common
macroalga by several authors (e.g. Hieber & al., 2001; Canto-
nati & al., 2006; Krizmanic & al., 2008). In Spain, H. foetidus
has been reported in the Pyrenees (NE Spain) (Margalef, 1948;
Llimona & al.,1985), Cantabrian mountains (N Spain) (Mar-
galef, 1950) and Sierra Nevada (Granada, S Spain) (Sánchez-
Castillo, 1984) with this study being the first recordin for the
Castilla-La Mancha region. As for habitat conditions, H. foe-
tidus is clearly rheophylic and stenothermic, disappearing when
water temperature rises above 10 ºC or 15 ºC according to Wehr
& Sheath (2003) and Starmach (1985), respectively. A combi-
nation of hydrological factors and light conditions controls the
seasonal fluctuations of this macroalga (Cantonati & al., 2006).
In general, this species is commonly found during winter and
spring (Hieber & al., 2001). In Spain, it is a vernal species found
occasionally in summer at high altitudes (>2000 m) (Llimona &
al., 1985). Our material was collected in early spring at a high
altitude (1564 m) when water temperature was 7.8 ºC. Both the
cellular size (from 7 µm to 13.5 µm in length) and the thallus
length (up to 20 cm) of the specimens coincide with previous
literature which describes large specimens (up to 30 cm) with
cells measuring from 8 µm to 12 µm in length (Bourrelly, 1957;
Rodriguez & Vergon, 1996; Wehr & Sheath, 2003).
ACKNOWLEDGEMENTS
This work was supported by the Junta de Comunidades de Castilla-La
Mancha, Consejería de Educación y Ciencia, through two regional research
projects (PO1109-0190-8090 1349 and PPII10-0271-1349).
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Sheath, R.G., Vis, M.L. & Cole, K.M. 1993. Distribution and systematics of
Batrahcospermum (Batrachospermales, Rhodophyta) in North America. 3.
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Sladecek, V. 1973. System of water quality from the biological point of view.
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Starmach, K. 1977. Phaeophyta. Rhodophyta. Flora Stodkowodna Polslci, vol.
14. Polska Academia Nauk, Instytut Botaniki.Warsaw.
Tiwari, D.N. 1978. The heterocysts of the blue-green alga Nostochopsis loba-
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bución de Tetrasporidium javanicum Möbius, 1893 (Chlorophyta: Tetras-
porales) en la cuenca del río Ebro. Limnetica (in press).
Van Oye, P. & Hubert, B., 1937. Recherches sur les “crons” du jurassique bel-
ge. Données générales. Biologische Jaarboek Dodonaea 4: 231-235.
Vrhovsek, D., Kosi, G., Krivograd-Klemencic, A., Smolar-Zvanut, N., Bernik,
T., Grbovic, J., Roser-Drev, A. & Remec-Rekar, S. 2006. Monograph on
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Associate Editors: Javier Fuertes / Santos Cirujano
Received: 19-VI-2012
Accepted: 7-IV-2013
5.3. Sobre la presencia de Nostochopsis lobata Wood ex Bornet et Flahault
en España: aspectos morfológicos, ecológicos y biogeográficos.

Título original: On the presence of Nostochopsis lobata Wood ex Bornet

et Flahault in Spain: morphological, ecological and
biogeographical aspects
Autores: José Luis Moreno, Marina Aboal y Laura Monteagudo
Publicación: Publicado en Nova Hedwigia, 85(3-4).
Impact Factor: 0.809
DOI: 10.1127/0029-5035/2012/0052
Fecha 2012

RESUMEN

Durante el muestreo habitual en ríos de la región, se detectó un cianófito

considerado ‘raro’ a escala europea, Nostochopsis lobata Wood ex Bornet et Flahault,
siendo la primera cita publicada para España. El ejemplar se recolectó en el Arroyo
Nava del Rey, un arroyo temporal que discurre sobre rocas metamórficas en una
zona de altitud media. En este trabajo se compara la morfología y ecología
descrita para la especie en diferentes regiones y se revisa su distribución
biogeográfica. En cuanto a morfología y preferencias de hábitat, el ejemplar
encontrado en el Arroyo Nava del Rey resultó ser similar a aquellos especímenes
detectados en Europa (Francia, Italia y Portugal). Sin embargo, difería de aquellos
descritos en zonas no europeas. En cuanto a su distribución geográfica, la revisión
de la bibliografía y de la base de datos de Biodiversidad Global (GBIF) reveló
que N. lobata es una especie cosmopolita, ya que se ha detectado en todas
las zonas climáticas a excepción de los polos (tropical templada, árida y fría).
Sin embargo, más allá de las zonas tropicales ha aparecido de forma mucho
más ocasional en el tiempo y en el espacio, habiendo sido detectada en la
mayoría de los casos una única vez. Además, cabe destacar que en las zonas
templadas la abundancia y tamaño de las colonias suele ser mucho menor, lo que
las hace más difíciles de detectar en campo. Un estudio ecológico y genético más
profundo permitiría comprender mejor la variabilidad de esta especie.

53
54
 Nova Hedwigia Vol. 95 issue 3–4, 373–390 Article
published online September 7, 2012

On the presence of Nostochopsis lobata Wood ex Bornet

et Flahault in Spain: morphological, ecological and
biogeographical aspects

Jose Luis Moreno1*, Marina Aboal2 and Laura Monteagudo1

1
Centro Regional de Estudios del Agua (CREA), Universidad de Castilla-La Mancha,
Crtra. de las Peñas, km3, Albacete-02071, Spain
2
Departamento de Biología Vegetal, Universidad de Murcia, Campus Espinardo, Murcia,
Spain

With 5 figures and 2 tables

Moreno, J.L., M. Aboal & L. Monteagudo 2012: On the presence of Nostochopsis lobata Wood
ex Bornet et Flahault in Spain: morphological, ecological and biogeographical aspects. – Nova
Hedwigia 95: 373–390.
Abstract: During a general biological assessment in rivers of southern Spain, a rare Cyanophyte
Nostochopsis lobata Wood ex Bornet et Flahault was detected for the first time in this country.
The species was collected in a temporary stream located in low altitude metamorphic mountains
(Nava del Rey stream). Morphological comparisons, as well as an ecological and biogeographical
review of the species are provided in this work. N. lobata from Spain shares similar morphology
and habitat preferences to specimens previously reported from Europe (France, Italy and Portugal).
However, when compared with specimens collected from non European sites, differences were
detected. In terms of geographical distribution, this study revealed that N. lobata is a cosmopolitan
species, distributed worldwide throughout tropical, temperate, arid and cold climate zones, with
the exception of the polar regions. The presence of this species in cold and dry climates was most
restricted. However, outside the tropical climate zone, the species shows a very sparse occurrence in
space and time, being collected in the majority of cases only once per site. Additionally, in temperate
climates, the colony size is rather small, the abundance is much lower and the colonies are very hard
to detect in the field. Further research on ecological and genetic aspects is needed in order to obtain
comprehensive knowledge of this variable species.
Key words: Blue-green algae, biogeography, environmental conditions, temporary stream, tropical species.

Introduction

The Cyanophyte genus Nostochopsis Wood ex Bornet et Flahault is included in the

Nostochopsaceae family (Bourrelly 1985, Komárek et al. 2003) and, like most of the

*Corresponding author: Jose Luis Moreno; e-mail: joseluis.moreno@uclm.es

© 2012 J. Cramer in Gebr. Borntraeger Verlagsbuchhandlung, Stuttgart, www.borntraeger-cramer.de

Germany. DOI: 10.1127/0029-5035/2012/0052 0029-5035/2012/0052 $ 4.50

55
eschweizerbart_xxx
genera of Stigonematales, is monospecific (Anagnostidis & Komárek 1990). A number
of other Nostochopsis species have been described, including N. radians described
by Bhâradwâja (1934) and also reported by Desikachary (1959) and N. wichmannii
Weber van Bosse 1913 (Yoneda 1939, Hirano & Hirose 1977). However these species
have subsequently been accepted as synonyms of N. lobata.
Nostochopsis lobata was first described in 1869 by Wood (in Wood 1872) in the
temperate climatic zone around Philadelphia, though traditionally it has been mainly
considered a tropical species rarely found in temperate zones (Frémy & Feldmann
1934, Sarma & Chapman 1975, Anagnostidis and Komárek 1990). In fact, it is widely
distributed in tropical areas worldwide, including Africa (Frémy 1929), South America
(Cáceres 1973, Branco et al. 2001), Asia (Geitler 1930-1932, Watanabe & Komárek
1988, Peerapornpisal et al. 2006, Aziz 2008) and Oceania (Sarma & Chapman 1975,
Sheath & Cole 1996). However, the number of records of N. lobata in climatic zones
other than the tropics has increased in recent decades. Thus, according to Komárek
(2003) the species is distributed within temperate, subtropical and tropical regions, but it
is mainly confined within the tropics and subtropics (Anagnostidis and Komárek 1990).
In Europe, there are only two published records of Nostochopsis lobata previous
to this study: one from Banyuls sur Mer in France (Frémy & Feldmann 1934) and
the other from L’Aquila in Italy (Del Grosso 1975). Hansgirg (1892) reported this
species in Bohemia (Czechoslovakia) under the name of Nostochopsis lobatus var.
stagnalis, but some authors have doubted the validity of this identification (Frémy
and Feldmann 1934, Bourrely 1985, Anagnostidis and Komárek 1990). It might be
noted that Hoffmann (1990) found a specimen in Corsica which had originally been
assigned to Mastigocladopsis jogensis but subsequently identified as Nostochopsis sp.
(Gugger & Hoffmann 2004). In addition, The Portuguese Algal Collection includes a
strain of N. lobata (http://www.gbif.org/) but no publications describing this finding
have been released.
During a general monitoring study in rivers of south-central Spain, several specimens
of N. lobata were found among the material collected in a temporary stream, Nava
del Rey stream. The main objective of this study was to review and compare the
morphology, ecological conditions and geographical distribution of this rare species.

Materials and methods

Sampling and processing: Since the year 2001 more than three hundred stream reaches in central
Spain have been surveyed repeatedly for aquatic macrophytes (i.e. submerged and floating aquatic
vegetation including macroalgae, bryophytes, pteridophytes and phanerogams). The stream reaches
surveyed were 100m long. Macrophytes were collected by hand and fixed in the field with 4% formalin.
This material was examined in the laboratory under a Leica M165C stereoscope and an Olympus
BX50 light microscope, both equipped with a Leica DFC420C digital camera. Measurements and
photomicrographs were taken and Glycerin-gelatin was used to make permanent slides.
Physico-chemical parameters: Physico-chemical analyses were performed and environmental
conditions in the Nava del Rey stream were measured on four dates in order to cover seasonal changes
in the aquatic environment (Table 1). Water flow (discharge) was calculated by integrating depth
and current velocity sections (Platts et al., 1983) measured at regular distances along three random
transects, and using a MiniAir© current meter. Electrical conductivity, pH and dissolved oxygen

56
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were measured in situ using appropriate sensors (Multiline P4 WTW). Alkalinity was also obtained
in field using the sulphuric acid method (APHA, 1998). One water sample per date was collected in
polyethylene bottles (500 ml) after filtration using a syringe with a Swinnex filter holder (Millipore
0.7 µm) and was kept in the refrigerator at 4°C for analysis, always within 48 h after sampling.
Nutrient concentration (N-NO3-, N-NO2-, N-NH4+, and P-PO4­-3) was determined photometrically with
MERCK kits (Spectroquant©). Detection levels for nutrients were: 0.5 mg L-1 for nitrate, 0.05 mg L-1
for nitrite and 0.03 mg L-1 for ammonium and phosphate. In the case of nitrate analysed on the last
date (Table 1), the detection level was decreased to 0.01 mg L-1 performing the cadmium reduction
method (APHA, 1998). Ion chromatography was used to analyse chloride and sulphate; while total
organic carbon (TOC) and inorganic carbon (IC) were analysed using infra-red spectrophotometry;
turbidity with a turbidimeter TN-100 (Eutech Instruments; infrared light); calcium and magnesium
by complexometry (volumetry); and sodium and potassium by atomic emission. All these parameters
were analysed following standard procedures detailed in APHA (1998).
Biogeographycal approach: For the biogeographical study of this species, references included in
both the scientific literature and the Global Biodiversity Information Facility (GBIF) were compiled
and a data base was created. Finally, this data set was combined in ArcMap (ESRI), with updated
Köppen-Geiger climate classification maps provided by the Food and Agriculture Organization of the
United Nations (http://www.fao.org/sd/EIdirect/climate/EIsp0002.htm) and by Kottek et al. (2006),
for the periods 1951 to 1976 and 1976 to 2000.

Results

Study site, ecological conditions and habitat: N. lobata was found among the material
collected in the Nava del Rey stream sampling site (Fig. 1) (UTM: X = 429700, Y
= 4265000) at medium altitude (590 m) (Fig. 2), a temporary warm stream located
in the southern part of the Iberian Peninsula (Sierra Morena mountains, province of
Ciudad Real, Autonomic Community of Castilla-La Mancha). On average, the stream
was 3 m wide and 20 cm deep . The sampling site was located 9.5 km from the source
(2nd order; upstream drainage area: 41.76 km2). The stream is a tributary of one of the
largest rivers in the Iberian Peninsula, the Guadalquivir River. This stream runs over
Palaeozoic siliceous metamorphic rocks (quartzites, slates and shales) (González &
Vázquez 2000). The climate is Mediterranean semiarid with a mean annual precipitation
of 600 mm (Fernández 2000). The main land use is extensive farming for cattle, and
the landscape corresponds to Spanish "dehesa", a kind of managed Mediterranean
maquia with Quercus ilex L. as the main tree species. The percentages of land uses
in the drainage area were 81.23% for forestry, 18.62% for dry land agriculture and
0.15% for urban use.
Spring-summer water temperature ranged from 18–27°C, which is typical of warm
water, Mediterranean streams located at low altitudes in a semiarid climate. Discharge
was low (0–70 L s-1) showing a seasonal flow pattern which varied from high spring
flow to summer isolated pools; the wet stream bed ranged from 1 m to 6 m wide. Phy-
sico-chemical characteristics of water included low conductivity values around 200 µS
cm-1 due to the presence of siliceous rocks and pH values around neutral or slightly
basic. According to its nutrient load, the water can be classified as oligo-mesotrophic
waters (Table 1).
Several specimens of N. lobata were found attached as epiphytes on a dead stem of
moss as well as free floating fragments entangled with other macrophytes. The species

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Table 1. Environmental features of the Nava del Rey Stream (Central Spain) on four dates. "-" no data.

Sampling date 09/06/2009 28/04/2010 05/08/2010 13/06/2011

Nostochopsis lobata present not detected not detected not detected

flow conditions base flow high flow isolated pools base flow

Field measurements (in situ)

wet channel width range (m) 1–4 1.5–6 1–3 1–5

discharge (L s-1) 30 70 0 40

conductivity 20º (µS cm-1) 237 192.70 262 180.8

dissolved oxygen (mg L-1) 6.84 9.11 6.41 10.16

dissolved oxygen (% 82.6 117.50 87.3 142.5
saturation)
pH 7,47 8.30 7.27 9.33

T (ºC) 18.8 24.50 26.5 29.1

total alkalinity (mg L-1) - 104.00 92 104.0

Laboratory analysis

ammonium (N-NH4+ mg L-1) - < 0.03 0.03 0.042

nitrite (N-NO2- mg L-1) - < 0.05 0.063 0.004

nitrate (N-NO3- mg L-1) - < 0.50 0.34 0.33

ortophosphate
- - 0.198 0.001
(P-PO4-3 mg L-1)
chloride (mg L-1) - 15.44 - 12.19

sulphate (mg L-1) - 41.50 - 30.17

potassium (mg L-1) - 1.27 - 2.02

sodium (mg L-1) - 8.57 - 8.67

calcium (mg L-1) - 23.00 - 25.97

magnesium (mg L-1) - 10.00 - 8.08

TOC (C mg L-1) - 2.44 - 7.69

turbidity (UNF) - 2.79 0.48 1.58

IC (C mg L-1) - 9.26 - 9.83

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Accompanying aquatic flora Ranunculus
R. peltatus R. peltatus R. peltatus
(macrophytes) peltatus
Tetrasporidium
Hydrodyction
Cladophora sp. javanicum H. reticulatum
reticulatum
Möbius
Spirogyra sp. Cladophora sp. Cladophora sp. Cladophora sp.

Zygnema sp. Spirogyra sp. Oedogonium sp. Nostoc sp.

Oedogonium Stigeoclonium
Spirogyra sp. Spirogyra sp.
sp. sp.
Stigeoclonium
Oedogonium sp. Zygnema sp.
sp.
Melosira sp. Anabaena sp.

Oscillatoria sp.

was associated with aquatic plants living in depositional habitats of pools or stream
banks. Abundance in the field was very low since the species was not detected with
the naked eye and only one small stem with Nostochopsis was found within a 100 m
stream reach.

Associated flora: Nostochopsis lobata was found only in June 2009, when the stream
showed its typical base flow conditions. The associated aquatic plants and macroalgae
were represented mainly by Cladophora sp. mats along with other filamentous green
algae such as Zygnematales and Oedogonium sp., and also irregularly dispersed stands
of Ranunculus peltatus Shrank (Table 1). In April 2010 discharge was the highest,
corresponding to spring flow, but N. lobata was not detected despite the high diversity
of the aquatic vegetation. In August 2010 the stream was a mosaic of isolated pools
nearly chocked with high biomass of chlorophytes Hydrodyction reticulatum (L.) Bory,
Oedogonium sp. and Zygnematales, with colonies of Cyanophytes (Anabaena sp.,
Nostoc verrucosum (Vaucher) Bornet & Flahault) and stands of Ranunculus peltatus
also dispersed along shallow pools. N. lobata was not recorded. Finally, we returned to
the site in June 2011 (the same month that it had previously appeared) but the species
was not detected (Table 1). In this last date the stream showed the base flow conditions
with Cladophora sp. and Zygnematales as the predominant aquatic flora.

Morphology: Several compact young colonies were found attached to a small dead
moss stem while numerous senescent fragments were found free floating in the
sample. Young colonies were globular and irregularly lobed, measuring 0.5–3 cm in
diameter (Table 2, Figs 3a, 3b and 4a). Additionally, numerous gelatinous diffluent
fragments were found entangled with filamentous green algae such as Cladophora
sp. or Zygnematales, and also in stands of R. peltatus (Figs 3c and 3d). The size of
these fragments were smaller than young colonies, measuring around 0.1–1 cm. The
colour of the colonies after formalin fixation was rusty-brownish in young specimens,
due to ferric precipitates, and bluish in the case of free floating fragments (Fig. 3a,
3b, 3c and 3d).

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Fig. 1. Location of study site: Nava del Rey Stream, southern Spain. In Guadalquivir River, water
flows from east to west.

In young colonies as well as in senescent fragments, filaments were uniseriate with

true branches and were embedded in mucilage which contained occasional filaments
of Calothrix fusca Bornet.
Two different regions were clearly observed in transversal section of young colonies
(Fig. 4b). The internal region was characterized by an intricate mass of main trichomes
(Fig. 3e) from which several branches emerged. Main trichomes were composed of
barrel-shaped and sometimes torulose cells measuring from 1.8–4 µm in diameter
and from 1.9–8.3 µm in length (Table 2; Figs 4c and 4d). In this region, branches
were sinuous to spiral in arrangement, formed by cylindrical cells measuring from
1.4–2.9 µm in diameter and from 4.6–12 µm in length (Table 2). The layout of main
trichomes was gradually radial in the external region of young colonies. In this case,
cells composing main trichomes were slightly smaller (from 1.6–3.4 µm in diameter
and from 2.7–6.9 µm in length) than those in the internal region but similar in shape
(Table 2). The same pattern was observed when comparing cells from external branches
with those from internal branches, the cell shape was similar but a decrease in size
was observed, particularly in cell length, since measurement of cells from external
branches was 1.3–2.6 µm in diameter and from 2.7–8.1 µm in length (Table 2). The
layout of branches in the external region was slightly sinuous to straight (Fig. 3f). In
both regions, T- type branching was predominant although some reverse V-shaped
branches were also observed. Short branches frequently formed a right angle with main
trichomes (Fig. 4d), while longer branches often formed an acute angle (Fig. 4c). Some
variability in filament morphology was noted irrespective of the position inside the
colony. On some occasions main trichomes, and branches which emerged from them,

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Fig. 2. Study site: Nava del Rey Stream. a) General view. b) Habitat view.

were formed by almost isodiametrical cells measuring from 1.5–2.5 µm in diameter

(Fig. 4d), probably representing the young and actively growing state of filaments.
In senescent fragments, the cell-type of main trichomes was torulose and slightly
elongated with a diameter range from 2.3–6.2 µm and a mean length of 7.2 µm (Fig. 4c).
Branches were composed of cylindrical cells, longer than in young colonies, measuring
up to 13 µm, with a diameter/length ratio of 1:5.
In both young colonies and senescent fragments, filaments were gradually narrowed
to the tip with end cells pointed, barrel or clavate shaped (Fig. 4f). Wide individual
hyaline sheaths were clearly visible especially near the outer margin of the colony (Fig.
4e). Heterocytes were colourless and spherical to conical in shape, ranging from 3.5 µm–
8 µm in diameter and 3.2 µm–8 µm in length, being slightly bigger in senescent than
in young specimens and more abundant in the central region. Intercalary heterocytes
were rare (< 5%) while lateral ones were frequent. Both being sessile and stalked
heterocytes were present (at the end of 1 to 4 celled branches) (Fig. 4c).
Hormogonia were observed in the outer colony regions and were more abundant in
senescent fragments. They comprised 2 to 10 isodiametrical cells which resulted from
filament fragmentation (1:1 diameter/length ratio) (Fig. 4g).

Biogeography: The combined distributional records show that N. lobata has a largely
worldwide distribution (Fig. 5). From a total of 96 sites included in the dataset, 52
(54%) were located within the tropics and 44 (45%) elsewhere. The northernmost
and southernmost sites were located in Canada (53°35'12.04"N) and New Zealand
(36°51'26"S) respectively.
In terms of the Köppen-Geiger climatic zones, N. lobata has been recorded in 4 of the
5 main climate zones: tropical, temperate, dry and cold. Polar is the only climate in
which this species has never been reported. As shown in Fig. 5, 89% of sites included
in this study corresponded with tropical (46%) and temperate (42%) climates while
only 10% corresponded with dry (5%) and cold (5%) climates. N. lobata was found in

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Table 2. Morphological description and ecological observations of Nostochopsis lobata from Spain
and other European specimens.

Reference and finding This study

location Ciudad Real, Spain
Life form senescent colony: young colony:
soft laminar solid, irregularly lobed, becoming hollow as it
fragments gets older; colour: brownish-rusty due to ferric
precipitate
Colony diameter (cm) - 0.1–3

Main trichomes external zone internal zone

layout tangled radial tangled

cell shape torulose barrel-shaped barrel-shaped

cell diameter (µm) 2.3–6.2(3.4) 1.6–3.4(2.3) 1.8–4(2.6)

cell length (µm) 4–13(7.2) 2.7–6.9(4.3) 1.91–8.3(5.1)

cell ratio diameter:length 1:2.4 1:1.8 1:2

Branches T- and V- branching

branches layout sinuous to spiral straight- sinuous-spiral

sinuous
cell form cylindrical cylindrical cylindrical

cell diameter (µm) 1.7–3.1(2.3) 1.3–2.6(1.8) 1.4–2.9(2)

cell length (µm) 7–16(11.2) 2.7–8.1(5.5) 4.6–12(8)

cell diameter:length 1:5 1:3 1:4

Trichomes ending gradually narrowed to the tip

Heterocytes lateral, sessile and stalked; intercalary less than 5%; spherical to
conical; colourless

size (µm) 4–8(6.7) wide; 3.5–6.5(5) wide;

5–8(7.2) long 3.2–7.2(5) long
Sheath hyaline, sometimes visible, especially in the outer margin.
7–10 µm wide
Mucilage (endophytic containing Calothrix fusca.
species)
Hormogonia 2–10 isodiametrical celled branches
size (µm) 2.2–3.2 wide , 2.2–3.9 long
Ecological observations sampling date: 9 June;
water conditions: T = 26.5°C, pH = 7.27, oligo-mesotrophic waters;
free floating (senescent) and epiphyte (young);
accompanying flora: filamentous green algae;
slow water shallow stream
siliceous metamorphic rocks (shales, quartzites, slates)

62
eschweizerbart_xxx
Frémy & Feldmann, 1934 Del Grosso, 1978 Hoffman, 1990
Banyuls sur Mer, France L’Aquila, Italy Corsica, France
solid globular thallus becoming spherical, subspherical or lobed similar to Nostoc or Rivularia;
hollow when adult; colonies; hollow and irregularly lobed;
colour: green-brownish colour: green-brownish colour: blue-green
1–1.5 3–5 maximum 4

external zone internal external zone internal -

zone zone
radial tangled radial tangled -

cylindri- isodiamtetrical cylindri- torulose; barrel-shaped to

cal cal cylindrical
2–6 3–5 8 4–5 -

10–12 8 9–11 -

1:1 1:2.75 1:1 1:2 -

T- and V- branching T- branching T- and V- branching

sinuous - spiral helicoidal -

cylindrical cylindrical cylindrical

2–3 2 2.0–4.8

10 12 2.6–18.0

1:5 1:6 -

gradually narrowed to the tip gradually narrowed to the tip tapering and slightly pointed.

lateral, sessile and stalked; lateral, sessile and stalked;

- intercalary rare; spherical to intercalary less than 5%;
elongated; colourless spherical to ovate
6–8 diameter 8 diameter 6.2–9.6 wide and 5.4–12.0
long
pseudosheath and true sheath - hyaline, sometimes visible.

containing Calothrix fusca - -

- 9–18 celled branches barrel-shaped cells branches

sampling date: 1 July;
water temperature = 21°C;
associated to Nostoc linkia;
attached to submerged rocks;
slow water;
siliceous metamorphic rocks
(shales)
sampling date: 25 August;
water conditions: T = 13°C,
pH = 7.4;
associated with Cladophora
glomerata;
found at 1m depth in slow
water calcareous stream
3.4–4.2 wide and 2.4–3.8 long
sampling date: September;
growing together with
Hildenbrandia rivularis;
attached to submerged rocks;
granitic region

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eschweizerbart_xxx
Fig. 3. Some images of Nostochopsis lobata. a) Habitus of a young colony. b) Fragment of a young
colony. c) Fragments of a senescent colony. d) Young colony (left) and senescent fragments (right).
e) General view of the inner of a young colony. f) Outer margin showing the layout of filaments
(young colony).

15 of the 31 Köppen-Geiger sub-climatic classes, which were defined by temperature

and precipitation conditions (Kottek et al 2006). Surprisingly, among the sites included
in this study, the sub-climate with the highest percentage of appearance was temperate-
fully humid with hot summers (25.00%) followed by tropical-winter dry (19.79%) and

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Fig. 4. Nostochopsis lobata. a) Macroscopic view of a young colony; arrows represent the union
point with a dead moss stem. b) Transversal section of a young colony; discontinuous line separates
internal (tangled) and external (radial) layout of filaments. c) Mature filament with sessile and stalked
heterocytes; branches are mostly in acute angle with main trichome. d) Filament in active growing;
branches are in right angle with main trichome. e) Filaments with visible sheaths and formation of a
T-type branch. Arrow indicates the septation of the branch-point cell. f) Different ends of filaments.
g) Hormogones. Scale bar represents: 1500 µm for a); 500 µm for b); 20µm for c) and d); 5 µm for
e), f) and g).

tropical-fully humid (13.54%). N. lobata has been detected in all tropical sub-climate
conditions. However, it has never been detected in 3 of the 9 temperate sub-climates:
fully humid with cool summer, winter dry with cool summer and summer dry with

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Fig. 5. World distribution of Nostochopsis lobata throughout main climates (Köppen-Geiger
classification system): Tropical (black), Temperate (medium gray), Arid (light gray), Cold (striped)
and Polar (white). Circles represent records supported by scientific publications; squares represent
data obtained from the Global Biodiversity Information Facility (GBIF).

cool summer. Thus, temperate sub-climates in which N. lobata has not been detected
are characterized by both cool summer and cold winter.
The presence of this species in cold and dry climates was the most restricted. Only
five references have been found corresponding with both climates. All cold-climate
sites were located in North America while dry-climate sites were distributed among
North America, Africa and Europe.
Focusing its European distribution, N. lobata has been referred to from two different
climates: temperate (France, Italy, and Portugal) and arid (Spain). In the case of Spain,
the site was located in a sub-climate zone currently designated as steppe-cold arid (for
the 1972–2000 period), thus constituting the first European arid zone in which N. lobata
has been detected. However, it should be noted that a recent change in the climatic
conditions in the Nava del Rey catchment was detected. The climate for the period
1950–1975 was classified as temperate according to the Köpen-Geiger classification.

Discussion

Morphology: Bhâradwâja (1934) described Nostochopsis radians as a new species of

Nostochopsis differing from N. lobata by a much smaller, solid thallus, in the absence
of special unseptate branches, and in the ends of the ultimate branches which were
not club-shaped or clavate. Most recently, N. radians was reported from the Hawaiian
Islands by Filkin et al (2003). These authors found both N. lobata and N. radians

66
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growing in distant streams. They differentiated the two species by virtue of N. lobata
colonies having hollow interiors, and increased branching towards the tip. Whereas N.
radians appeared as a solid mat with branching that did not increase towards the tip.
In terms of environmental conditions, both streams were similar in pH (8.1 and 8.5
respectively) and water temperature (22°C, 24°C respectively) although the stream
where N. lobata was found was wider, deeper and with lower water conductivity
(90 µS cm-1) than the stream with presence of N. radians (200 µS cm-1). Thus, even
though this species is accepted by some authors (Desikachary 1959), differences
between N. radians and N. lobata were not consistent enough to be considered a
separate species, possibly due to the variability existing among different growth stages
and environmental adaptations of Nostochopsis (Frémy and Feldmann 1934).
Comparing our material with N. lobata specimens described worldwide, the greatest
differences were found with those from New Zealand (Sarma & Chapman 1975),
Australia (Skinner & Entwisle 2001), Thailand (Peerapornpisal et al 2006) and
Bangladesh (Aziz 2008) with respect to heterocytes and branching. Sarma & Chapman
(1975) and Aziz (2008) described branches formed by cells measuring up to 18 µm
in length and emerging unilaterally whereas in the present study slightly smaller
cells and both-direction branching were observed. Heterocytes described by Sarma
& Chapman (1975) and Skinner & Entwisle (2001) showed a wide range of sizes
reaching up to 12 µm in length, although such large heterocytes were not observed in
the Spanish material from Nava del Rey. In contrast, Peerapornpisal (2006) described
smaller spherical heterocytes ranging from 3–4 µm in diameter. According to Frémy &
Feldmann (1934), Del Grosso (1975), Tiwari (1978) and Hoffmann (1990), intercalary
heterocytes were rare, rounded and situated in main trichomes while Cáceres (1973) and
Skinner & Entwisle (2001) described frequent intercalary heterocytes characterized by
a cylindrical shape. Nevertheless, variability in frequency of intercalary heterocytes has
been related with nitrogen content in culture (Tiwari 1978) with intercalary heterocytes
suppressed in the presence of ammonium.
Some discrepancies were found with respect to branching type. Various authors
described T-branching ( Del Grosso 1978, Skinner & Entwisle 2001) while others also
mentioned reverse V-shaped branching (Frémy and Feldmann 1934, Komárek 2003).
However, a recent study by Gugger & Hoffmann (2004) revealed that V- or Y- shaped
branches in Nostochopsis start out as typical T-branches and subsequent modifications in
the branch-point cell could lead to a final V- or Y-shaped appearance. This observation
supports the idea proposed by Begum et al. (1994) that Mastigocladopsis jogensis is a
synonym of N. lobata since reverse V-shaped branching is not a stable characteristic
and could be formed at a late stage of growth.
A further comparison of our material with the other specimens collected within
Europe (Frémy & Feldmann 1934, Del Grosso 1978, Hoffmann 1990) revealed few
morphological differences. For instance, cells composing the main trichomes and
branches in young colonies seemed to be slightly smaller and less elongated than those
described by Frémy & Feldmann (1934) and Del Grosso (1978) while the opposite
occurred for cells from senescent fragments (Table 2). Additionally, our material showed
wider size ranges than other European specimens. This may be due to differences in

67
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age and growth stage among the specimens observed. Frémy and Feldmann (1934),
observed two different filament morphologies (Figs 4c and 4d). Filaments highly
branched, formed by small isodiametrical cells (Fig. 4 d) which were observed forming
spreading in small groups among young colonies. Therefore, these regions could be
zones of active growth, supporting the hypothesis of Cáceres (1973) that growth in
this species is produced by a lateral increase in the number of trichomes and not by
centrifugal growth.

Habitat and environmental conditions: In terms of habitat, N. lobata shows a high

degree of variability. It has been encountered in lakes (Tiwari 1977, El Saied 2007),
running waters (Frémy & Feldmann 1934, Palmer 1941, Pandey & Pandey 2008),
thermal springs (Yoneda 1939), as an epiphyte (Sarma & Chapman 1975, this study),
attached to rocks (Hoffmann 1990, Peerapornpisal 2006), free floating (Wood 1872), on
moist soils (Skinner & Entwisle 2001, Aziz 2008) and even on humid walls (Skinner &
Entwisle 2001) among others. However the most extreme habitat where N. lobata has
been found is probably as cryptoendolithic in an arid climate (Weber et al 1996). These
authors found this species in South Africa growing together with Chroococcidiopsis
sp. in the pore spaces inside sandstones. This is the only place where N. lobata has
been found growing as cryptoendolithic and the strain isolated by the authors (SAG
2.97) has been used in geobiological (Büdel 1999, Büdel et al 2004) and genetic
studies (Gugger & Hoffmann 2004). In this case, the polyphyly of true branching
cyanophytes was confirmed as well as the possible need for including Westiellopsis,
Fischerella and Nostochopsis in the same genus, due to sequence similarity above
95%. These genera show great morphological variability depending on growth stage
and environmental conditions, thus the differentiation of these genera by means of
traditional alpha taxonomy, based mainly on morphological characteristics, can lead
to possible misidentifications. Polyphasic studies are necessary in order to avoid this
problem. Additionally, genetic analyses are also necessary to test the monospecific
character of the genus Nostochopsis.
Focusing on European reports, there are some similarities in environmental conditions
of sites: (1) in all cases the collection took place in summer in shallow warm waters,
(2) all streams run over siliceous metamorphic rocks, except the Tirino stream (Italy)
(Table 2). Thus, the underlying lithology could be a very important factor conditioning
the presence of N. lobata. This is supported by the work of Branco et al. (2001) in
Sao Paulo State (Brazil) who suggested that N .lobata shows a preference for low
conductivity waters, although further research on ecological preferences of this
species is required.
With respect to the environmental conditions in Nava del Rey stream, N. lobata was not
found in three additional site visits (Table 1). Comparing the two dates in June, some
differences in pH, temperature, dissolved oxygen and conductivity were apparent. A
narrow range of these ecological factors may define the restricted ecological niche where
this species grows and, therefore, explain why this species and other Stigonematales
are absent most of the time from its type localities (Hoffmann 1996). We can conclude
that only occasionally the studied stream achieves the restricted ecological conditions
required by N. lobata, during a short time in the summer.

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Finally, it must be highlighted that the interest in nutritive and pharmacological
properties of N. lobata is rapidly growing, especially in Asia (e.g. Peerapornpisal
2006, Pandey & Pandey 2008), which could increase the knowledge of this interesting
species.

Biogeography: In terms of geographic distribution, N. lobata can be considered as a

(sub-) cosmopolitan species since it is distributed worldwide, being found in tropical,
temperate, arid and cold climatic zones, i.e., in all climates except polar (Fig. 5).
However, some differences in form exists among climatic zones. For example, in
several tropical zones as well as adjacent temperate sub-climatic zones, N. lobata grows
as to macroscopic proportions (more than 10 cm after Peerapornpisal et al 2006) and
is locally abundantly, and used as a local food and medical resource (Peerapornpisal
et al 2006, Pandey & Pandey 2008). In contrast, in temperate zones, N. lobata grows
as small spherical colonies which are difficult to detect in the field. Some examples
include specimens from Philadelphia described by Wood (1872) measuring about
4 cm; Banyuls sur Mer (France) 1–1.5 cm according to Frémy & Feldmann (1934)
or those from New Zealand described by Sarma & Chapman (1975) measuring up
to 0.8 cm in diameter. This morphologic pattern is shown by some Cyanophytes
which have many specialized types restricted to small defined habitats and localities
but with morphologies that can be uniform in one area and variable in another
(Komárek 2003).
In the case of N. lobata from Spain, visits to the sampling site were repeated and the
species was not found again. Most of the stigonematalean genera are monospecific
and known only from their first discovered biotope (Anagnostidis & Komárek 1990).
Furthermore, it has been stated that majority of stigonematalean species have world-
wide distribution but usually occur in very limited and special conditions or under
limited ecological events, that is, in disjunct areas, in spite of their wide geographical
distribution (Anagnostidis & Komárek, 1990). For example, species associated
to restricted niches or habitats like thermal springs typically show a mosaic-like
geographical distribution reflecting the corresponding distribution of ecological
niches (Hoffmann 1996).
Thus, despite its worldwide distribution and its climatic tolerance, N. lobata may
show an underlying tendency: it grows abundantly in tropical regions where its
optimal climatic conditions are stable for several months whereas it grows scarcely in
other regions only when conditions allowing its germination and growth are reached,
probably for a few weeks in occasional years. This observation along with its small size
and low abundance could also explain why this species is hard to detect in temperate
sites where it has been previously found.

Acknowledgements

This study has been financed by Project PO1109-0190-8090 of the Ministry of Education and Science,
Autonomic Community of Castilla-La Mancha.

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Manuscript received September 15, 2012; accepted February 2, 2012.

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5.4. Eutrofización fluvial: Comparación del impacto del regadío y secano a
través de diferentes escalas espaciales.

Título original: River eutrophication: Irrigated vs. Non-irrigated

agriculture through different spatial scales
Autores: Laura Monteagudo, José Luis Moreno y Félix Picazo
Publicación: Publicado en Water Research, 46.
Impact Factor: 4.655
DOI: 10.1016/j.watres.2012.02.035
Fecha 2012

RESUMEN

En este trabajo se analizó la relación entre los diferentes usos de suelo y la

eutrofización de ríos y arroyos en la zona de estudio, haciendo especial hincapié en
conocer cómo afecta la elección de la escala espacial de estudio a los resultados. Se
calculó la composición porcentual de usos de suelo (agrícola, urbano y forestal) y se
midió la concentración de nitrato, amonio y ortofosfato en 130 puntos de muestreo
repartidos por la región. Además, para evaluar qué tipo de agricultura es
responsable en mayor medida del aumento de nutrientes en el agua, se dividió el uso
agrícola en tres subclases (regadío, secano y agricultura de bajo impacto).
Posteriormente se hicieron correlaciones entre la concentración de nutrientes en el
agua y los usos de suelo obtenidos a través de cuatro escalas espaciales diferentes:
dos amplias (área de drenaje total y corredores de 100m de anchura) y dos locales
(zonas de influencia de 1 y 5 kilómetros de radio). Los resultados mostraron que
utilizar diferentes escalas espaciales puede producir resultados diferentes y, por
tanto, puede llevar a conclusiones muy distintas. En el caso de escalas amplias, la
autocorrelación espacial y la representación insuficiente de algunos usos de suelo
produjeron resultados irreales. Por el contrario, con el uso de las escalas locales no
surgió el inconveniente de la autocorrelación y los usos de suelo estuvieron
representados de manera más uniforme. La escala de local de 1km radio aguas

73
arriba resultó ser la más apropiada para detectar eutrofización en el sistema fluvial
de Castilla-La Mancha, con el regadío como principal causante del aporte de nitrato
al agua. Además, se realizó un análisis de regresión segmentada mediante el cual se
detectó un umbral del 50% de regadío en la escala de 1km de radio. Este resultado
tiene una clara implicación en la gestión de la calidad del agua: para impedir que
aumente la eutrofización de ríos y arroyos es necesario evitar que haya zonas con
más de un 50% de regadío en 1 km de radio desde el cauce. Por último, se describió
una metodología base para elegir la escala espacial más apropiada en el estudio de la
eutrofización causada por contaminación difusa.

74
 w a t e r r e s e a r c h 4 6 ( 2 0 1 2 ) 2 7 5 9 e2 7 7 1

Available online at www.sciencedirect.com

journal homepage: www.elsevier.com/locate/watres

River eutrophication: Irrigated vs. non-irrigated agriculture

through different spatial scales

Laura Monteagudo a,*, José Luis Moreno a, Félix Picazo b

a
University of Castilla-La Mancha, Regional Center of Water Research (CREA), Crtra. de Las Peñas km3, Albacete 02071, Spain
b
University of Murcia, Department of Ecology and Hydrology, Murcia, Spain

article info abstract

Article history: The main objective of this study was to determine how spatial scale may affect the results
Received 5 October 2011 when relating land use to nutrient enrichment of rivers and, secondly, to investigate which
Received in revised form agricultural practices are more responsible for river eutrophication in the study area. Agri-
26 January 2012 culture was split into three subclasses (irrigated, non-irrigated and low-impact agriculture)
Accepted 16 February 2012 which were correlated to stream nutrient concentration on four spatial scales: large scale
Available online 28 February 2012 (drainage area of total subcatchment and 100 m wide subcatchment corridors) and local
scale (5 and 1 km radius buffers). Nitrate, ammonium and orthophosphate concentrations
Keywords: and land use composition (agriculture, urban and forest) were measured at 130 river reaches
Stream in south-central Spain during the 2001e2009 period. Results suggested that different spatial
Nitrate scales may lead to different conclusions. Spatial autocorrelation and the inadequate repre-
Land use sentation of some land uses produced unreal results on large scales. Conversely, local scales
Irrigation did not show data autocorrelation and agriculture subclasses were well represented. The
Catchment local scale of 1 km buffer was the most appropriate to detect river eutrophication in central
Spanish rivers, with irrigated cropland as the main cause of river pollution by nitrate. As
regards river management, a threshold of 50% irrigated cropland within a 1 km radius buffer
has been obtained using breakpoint regression analysis. This means that no more than 50%
of irrigation croplands should be allowed near river banks in order to avoid river eutrophi-
cation. Finally, a methodological approach is proposed to choose the appropriate spatial
scale when studying river eutrophication caused by diffuse pollution like agriculture.
ª 2012 Elsevier Ltd. All rights reserved.

1. Introduction proliferation of filamentous algae which leads to a decrease in

Land use transformation from natural to human dominated
systems has globally caused environmental impacts, making
land use analysis a useful indicator of changes in stream
ecosystems (Meyer and Turner, 1994). One of the most
extended impacts produced by landscape modification is
eutrophication of rivers, lakes and marine ecosystems (Lund,
1967; Omernik et al., 1981; Smith, 2003). Eutrophication
produces changes in community composition and the
dissolved oxygen levels, water quality, fish death and often to
a global loss of biodiversity (Carpenter et al., 1998; Quinn, 1991;
Smith et al., 1999). In order to plan effective prevention
measures against eutrophication, a deep understanding of the
relationship between land cover and stream conditions is
needed. However, this relationship is characterized by
numerous factors and environmental variables operating and
interacting through different scales, both spatial and temporal
(Frissell et al., 1986). Thus, several questions may arise when

* Corresponding author. Tel.: þ34 967599200x2576; fax: þ34 967599269.

E-mail address: laura.monteagudo@uclm.es (L. Monteagudo).
0043-1354/$ e see front matter ª 2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.watres.2012.02.035

75
2760 w a t e r r e s e a r c h 4 6 ( 2 0 1 2 ) 2 7 5 9 e2 7 7 1
assessing the relationship between stream conditions and
surrounding landscape. For instance: 1) which land use classes
may be analysed? 2) which spatial scale/s would be the most
appropriate (e.g. catchment, riparian buffers, circular buffers,
among others)? and 3) how may this spatial perspective
influence the interpretation of results? In this paper, we have
studied the influence of land uses on river nutrient concen-
tration focussing on different agricultural practices (Section
1.1). In addition, we have analysed how different spatial scales
may affect the results and how to identify which scale is the
most appropriate in this type of studies (Section 1.2).
1.1. Agricultural land use
Regarding the first issue, the land uses most commonly
studied as causes of river eutrophication have been both
agriculture and urban (Osborne and Wiley, 1988; Townsend
et al., 1997; Von Schiller et al., 2008). Whereas urban efflu-
ents cause point pollution, agriculture often causes the most
important source of non-point or diffuse pollution worldwide.
However, the level of eutrophication impact depends on the
kind of agriculture, for instance irrigated cropland vs. non-
irrigated. Irrigation allows crops to grow in permanently
water-scarce or temporarily water-stressed environments,
such as in Mediterranean countries and other semi-arid areas
worldwide (World Water Assessment Programme, 2009). Since
the increase in population and demand for food, along with
expected climate change over the next decades, suggest an
increase in irrigation surface and water demand in arid and
semi-arid areas (Tilman, 2001; World Water Assessment
Programme, 2009), the control of diffuse pollution in this
future scenario will be an important challenge, thus making
the identification of land uses responsible for freshwater
eutrophication a real need.
Few studies have been published comparing the eutro-
phication impact caused by different agricultural land use
classes. For example Johnson et al. (1997) compared rowcrop
vs. non-rowcrop agriculture in North America whereas
Lassaletta et al. (2009) compared arable lands, permanent
crops, pastures and heterogeneous areas in Spain.
According to the Spanish National Statistics Institute
(www.ine.es), irrigated agriculture is more intensive than
non-irrigated in terms of production due to the artificial
supply of water and fertilizers to croplands. This technique
increases the export of nitrogen compounds to ground- and
surface waters, making irrigated agriculture the main cause of
eutrophication in Spanish rivers (Álvarez-Cobelas et al., 2010;
Berzas et al., 2004; Cavero et al., 2003).
In this paper, we analyse the effect of some land uses
(agriculture, urban and forest) on river eutrophication, taking
into account three classes of agricultural land use with
different expected impact intensities: irrigated, non-irrigated
and low-impact agriculture.
1.2. Spatial scale
Regarding spatial scale, some authors have suggested that the
influences of land uses on stream ecosystems and water
quality must be analysed at the catchment scale (Omernik
et al., 1981; Richards and Host, 1994; Roth et al., 1996)
76
whereas others have argued that land uses located closer to
the stream (i.e. reach scale, local buffers, riparian corridors)
are more important (Harding et al., 1998; Nerbonne and
Vondracek, 2001). Recent studies suggest that a multiple
spatial scale approach could be a necessary step to identify the
appropriate framework (e.g. Chang, 2008; Tran et al., 2010). In
a multi-scale approach, different spatial scales would provide
different land use datasets and therefore different results. In
order to correctly interpret the results, some issues must be
taken into account to select the appropriate spatial scale: (1)
spatial autocorrelation between sites and (2) the lack of a wide
range of values (%) for each land use class. Autocorrelated
data violate the assumption of independence of most stan-
dard statistical procedures such as correlation analysis
(Legendre, 1993). In many cases, when working with large
scales (e.g. subcatchment drainage area or subcatchment
riparian corridors) autocorrelation is caused by overlapping of
subcatchment drainage areas of consecutive downstream
sites. The effect of overlapping on statistical analysis is also
known as pseudoreplication (Hurlbert, 1984) and the conse-
quence is an increase in the statistical power (higher corre-
lations than expected) but not in the biological or ecological
significance (Townsend et al., 1997).
The second issue is a consequence of working with land use
percentage values. Frequently, datasets include land use
classes that cover a wide range across the impact gradient (i.e.
sites with low, medium and high percentages of agriculture)
while other land uses (e.g. urban) only reach low percentages
within subcatchments (for example 10e20% as maximum). In
addition, as land uses are quantified in percentages of a surface,
the higher the percentage value for one land use, the lower
percentage value for the remaining land uses. The statistical
consequence is a masking effect of classes reaching low cover
percentage by those more extended in the subcatchment.
In this study, we relate land uses to nutrient concentration
in rivers through four different spatial scales (total drainage
area, 100 m wide corridors, and 5 and 1 km radius buffers
upstream sites) in order to identify the most appropriate spatial
framework. The novelty of this research lies in testing new
spatial scales (1 and 5 km upstream buffers) and comparing the
eutrophication effect caused by two kinds of agriculture
common in Mediterranean countries: irrigated vs. non-
irrigated croplands. Hence, the main aims are: (1) to demon-
strate that spatial scale affects the results of studies relating
land use to stream condition; (2) to determine which type of
agriculture (irrigated vs. non-irrigated) is more responsible for
river eutrophication and to define pressure thresholds (3) to
propose a method to select the most adequate spatial scale to
assess the eutrophication impact by diffuse pollution.
2. Material and methods
2.1. Study area
This study was carried out within the boundaries of the
Spanish administrative region of Castilla-La Mancha (south-
central Spain). The study area includes the upper and middle
reaches of five large river basins: Tajo, Guadiana, Guadalquivir,
Júcar and Segura. This region currently has the lowest
 w a t e r r e s e a r c h 4 6 ( 2 0 1 2 ) 2 7 5 9 e2 7 7 1
population density in Spain (26.4 inhab/km2). Industry has
noticeably increased during recent years but agriculture
continues to be the main economic activity. Land dedicated to
agriculture covers almost half of the entire surface of the region
(Ministerio de Medio Ambiente y Medio Rural y Marino, 2010),
most of which is dedicated to extensive non-irrigated cropland
(mainly cereals, vineyards and olive trees) (Table 1). Irrigated
cropland (mainly maize and sunflowers) is usually located
close to river banks. Due to water eutrophication by agricul-
ture, six zones within the study area have been designated by
law as “vulnerable” with regard to nitrate pollution (Consejerı́a
de Agricultura y Medio Ambiente, 1998, 2003; Consejerı́a de
Industria, Energı́a y Medio Ambiente, 2009). Despite this,
groundwater and river eutrophication continue to be a serious
problem for water quality in the study area (Álvarez-Cobelas
et al., 2010; Berzas et al., 2004; Moreno et al., 2006).
The climate in the study area is Mediterranean with
continental elements, exhibiting remarkable fluctuations in
daily and seasonal temperatures. Annual precipitation ranges
from 300 mm on the plains (semi-arid) to more than 1000 mm
in the mountains (humid), with rainfall mainly concentrated
in autumn (Fernández, 2000). As for geology, three zones can
be distinguished: the western zone is rich in Precambrian
siliceous rocks (mostly quartzite, slate, shale, granite and
gneiss); Mesozoic calcareous rocks (limestone, dolomite,
sandstone and conglomerates) are dominant in the eastern
area; and finally, Tertiary sedimentary fills are accumulated in
the great central plateau located at 700 m a.s.l. called “La
Mancha” where clays, sandstones, gravels, stones, conglom-
erates, marls and gypsum are abundant (González and
Vázquez, 2000). Mountains are located mainly on the edges
of the region and can reach 2000 m a.s.l.
2.2. Survey design
Sites were chosen in order to reflect the widest possible
gradient of land use and nutrient concentration. Nutrient
concentration was measured at 130 sites during the
2001e2009 period. In order to take seasonal differences into
account, samples were taken during two periods per year:
springesummer (SS) from March to September, and
autumnewinter (AW) from September to December. Some
sites subject to variations in nutrient concentration due to
human impact and/or seasonal conditions were visited more
than once. Thus, the total dataset was composed of 216 water
samples from 130 sites (Fig. 1), 148 were collected in the
Table 1 e Estimated land use composition in the study
area according to official data (Ministerio de Medio
Ambiente y Medio Rural y Marino, 2010).
km2
Total area 79,409
Urban 7941
Agriculture 36,530
Irrigated 5110
Non-irrigated 31,017
Forest 34,940
2761
springesummer season and 68 in autumnewinter season.
Sites were separated by a minimum distance of 5 km (Johnson
et al., 1997). This distance avoided subcatchment overlapping
when local scales were selected (see Section 2.4).
2.3. Chemical analyses
In order to compare different types of agriculture in terms of
eutrophication impact, dissolved inorganic nutrients (nitrate,
ammonium and phosphate e as soluble reactive phospho-
rous, SRP) were measured since these are the main nutrient
compounds released by the inorganic fertilizers applied to
crops. Water samples were taken in 250 ml polyethylene
bottles after filtration, and kept at 4  C in a cooler until anal-
yses. Nutrient concentration was determined photometrically
with MERCK kits (Spectroquant) within 48 h of sampling. The
values were expressed as mg L1 of N-NO þ 3
3 , N-NH4 and P-PO4 .
Detection levels were: 0.10 mg L1 for nitrate and 0.01 mg L1
for ammonium and phosphate. Confidence intervals for these
commercial kits were: 0.07 for phosphates, 0.06 for
ammonium and 0.3 for nitrate (http://photometry.merck.
de). In the case of nitrate it was necessary to decrease the
detection level for unpolluted sites by performing the
cadmium reduction method to reach the lowest detection
level of 0.01 mg L1 N-NO 3 (APHA, 1989).
2.4. Land use data
All analyses and geographic data management were carried
out with ArcGIS 9.2 (Esriª). Land use percentage composition
for each site was quantified with four different spatial scales:
total drainage area of subcatchment upstream from each site
(DA), 100 m wide subcatchment buffer corridors upstream
from each site (CD), and 1 and 5 km radius semicircular
buffers oriented upstream (B1 and B5 respectively) (Fig. 2).
The total upstream drainage area for each location (sub-
catchment drainage area) was delineated based on data and
images from the Spanish Spatial Data Infrastructure (IDEE;
http://www.idee.es/show.do?to¼pideep_desarrollador_wms.es).
Corridors were obtained by generating a 100 m wide buffer
along the stream drainage net upstream from each site.
Finally, buffers of 1 and 5 km radii were generated from each
site, but surfaces draining downstream sites (after intersect-
ing with DA polygons) were removed (Fig. 2). As mentioned in
Section 2.2, minimum distance between sites was 5 km. Thus,
only local scales (B1 and B5) failed to produce overlapping
areas on consecutive sites. Regarding the meaning of the term
“buffer”, in literature it usually refers to riparian corridors
created from longitudinal features (streams). However, here
we have used ArcGis terminology, which defines “buffers”
also as circular surfaces created around points (sites).
Land use data were obtained from the Corine Land Cover
% (CLC, 2000) Project. Land cover classes characterized in CLC
100 were reclassified into the following categories (Table 2): urban
10 and industrial land cover (URB); areas dedicated to agricultural
46 uses (AGR); and areas with natural or reforested vegetation
7 (FOR). Agricultural land use was also split into three classes:
39 irrigated cropland (IRR), non-irrigated cropland (NIR) and
44 agricultural land uses expected to have a low impact on river
ecosystems (LIA).
77
2762 w a t e r r e s e a r c h 4 6 ( 2 0 1 2 ) 2 7 5 9 e2 7 7 1
Fig. 1 e Site distribution map showing the entire study area and main basin boundaries. Black arrows show the flow
direction.
2.5. Statistical analyses
Statistical analyses were performed using STATISTICA v.9
(StatSoftª) and XLSTAT v.7.5.2 (Addinsoftª). Normality and
homogeneity of variance were tested using ShapiroeWilk and
Bartlett tests, respectively. These assumptions were not
satisfied and therefore, non-parametric analyses were
required. A KruskaleWallis test was applied to test statistical
differences between land use classes depending on the
selected spatial scale.
A preliminary interpretation of eutrophy trends was
carried out using correlation analysis between land use
percentages and dissolved nutrient concentrations,
a common statistical approach in this type of studies. In this
case, we calculated Spearman’s rank correlation coefficient
for each spatial scale dataset and for both springesummer
and autumnewinter seasons.
Spatial autocorrelation is defined as the property of
random variables which take values, at pairs of sites sepa-
rated a given distance, that are more similar (positive auto-
correlation) or less similar (negative autocorrelation) than
78
expected for randomly associated pairs of observations
(Legendre and Legendre, 1998). Autocorrelation indicates lack
of independence among the observations and creates prob-
lems when attempting to use statistical tests that require
independence of the observations. In order to test for spatial
autocorrelation in our data, we applied the simple Mantel test
which measures the correlation between two similarity or
dissimilarity matrices (rM(AB)): a matrix (A) of distances
computed for the environmental variable tested, and a matrix
(B) of geographic distances. Therefore, a correlation between
matrices A and B measures the extent to which the variations
in the similarities of A correspond to the variations in B. Thus,
similarity matrices containing the Euclidean distance
between observations were generated for the predictor vari-
able (land use composition through each spatial scale) and
also for the response variable (nutrient concentration). Each
matrix was then related with the geographic distance matrix
generated from UTM site coordinates.
A ManneWhitney test was used to test for differences in
nitrate concentration between two groups representing
different practices in agriculture: irrigated and non-irrigated
 w a t e r r e s e a r c h 4 6 ( 2 0 1 2 ) 2 7 5 9 e2 7 7 1 2763

Fig. 2 e Schematic representation of the spatial scales used in this study. Large scales: DA, total drainage area of upstream
subcatchment; CD, 100 m wide upstream subcatchment corridor. Local scales: B5, 5 km upstream radius buffer; B1, 1 km
upstream radius buffer. Subcatchment corridors in this figure are 500 m wide for an easier display.

cropland. Linear regression analyses between nitrate As for ammonium, 2.3% of the sites surpassed the maximum
concentration and both groups of agriculture were performed. toxic levels for aquatic life (around 2.5 mg L1; USEPA, 1999).
Finally, a breakpoint regression analysis was applied in order These values were recorded in the Tajo River below the joining
to identify any significant pressure threshold in the relation- of two tributaries collecting urban and industrial wastes from
ship nitrate-irrigated agriculture. For this purpose, we used the huge metropolitan area of Madrid. Nitrate concentration
SegReg software (Oosterbaan, 2010). never surpassed the limit established (50 mg L1) by the
Nitrate Directive (European Commission, 1991).
Results for ShapiroeWilk (W) test ranged from 0.347 to
3. Results 0.960 ( p < 0.001, a ¼ 0.05) suggesting that samples (% of each
land use class per scale) did not follow a normal distribution.
3.1. Exploratory analysis In addition, Bartlett’s test was significant (c2(23) ¼ 35.172,
p < 0.0001, a ¼ 0.05) and therefore, the null hypothesis of
Nutrient concentration ranged from <0.01 to 30 mg L1 for homogeneity of variances was rejected. Thus, the non-
nitrate (N-NO3 ), <0.01 to 5.5 mg L
1
for ammonium (N-NHþ 4) parametric Spearman’s rank correlation test between land
1
and <0.01 to 12.0 mg L for orthophosphate (P-PO34 ) (Table 3). use classes and nutrients was applied as a first exploration of
79
2764 w a t e r r e s e a r c h 4 6 ( 2 0 1 2 ) 2 7 5 9 e2 7 7 1
Table 2 e Description of land uses analysed in the present study with their corresponding Corine Land Cover classes.
Reclassification code Description
Urban Urban and industrial land use
Agriculture Areas dedicated to agricultural uses
Irrigated agriculture Including crops, fruit trees and
olive groves with a permanent
irrigation infrastructure
Non-irrigated Cereals, legumes, fodder crops,
agriculture root crops, fallow land, fruit trees
and olive groves without
irrigation infrastructures
Low-impact agriculture Pastures and abandoned agricultural
fields with dense grass cover and
not under a rotation system
Forest Areas with natural or
reforested vegetation
the relationship between land use and river eutrophication.
The value of the correlation coefficient varied clearly
depending on spatial scale selected and, to a lower extent, on
the season (Table 4). In general, positive and high correlation
values were obtained between agricultural land uses (AGR,
IRR, NIR) and nitrogen compounds, while phosphate concen-
trations were more correlated to urban land use percentage
(URB). The influence of spatial scale on the land use defined as
low-impact agriculture (LIA) was complex and difficult to
interpret, probably due to the heterogeneity of land uses
compiled in this class. For this reason, this land use class was
eliminated from statistical analyses. Natural or semi-natural
land cover classified as forest (FOR) was strongly and nega-
tively correlated with all nutrients irrespective of scale.
A global trend in relationship between nutrients and land
use was observed: for large scales correlation values were
higher than for local scales (Table 4). However, the relation-
ship between nitrate and irrigated cropland (IRR) showed the
opposite trend: the highest correlation values were observed
at 1 km buffer scale (r ¼ 0.451, p < 0.001) decreasing gradually
towards larger scales. Regarding the relationship nitrate-
irrigated cropland, the effect of seasonality was also remark-
able with higher values in springesummer, coinciding with
the irrigation season in the study area when nitrate leaching is
enhanced. However, for non-irrigated cropland, the highest
80
Corine Land Cover classes
1. Artificial areas
2. Agricultural areas
212. Permanently irrigated land
2212. Irrigated vineyards
2222. Irrigated fruit trees and berry plantations
2232. Irrigated olive groves
2412. Annual crops associated with permanent crops
on the same irrigated parcel
2422. Juxtaposition of irrigated small parcels of diverse
annual crops, pasture and/or permanent crops
2423. Juxtaposition of irrigated and non-irrigated small
parcels of diverse annual crops, pasture and/or
permanent crops
2432. Land principally occupied by irrigated agriculture,
with significant areas of natural vegetation
211. Non-irrigated arable land
2211. Non-irrigated vineyards
2221. Non-irrigated fruit trees and berry plantations
2231. Non-irrigated olive groves
2411. Annual crops associated with permanent
crops on the same non-irrigated parcel
2421. Juxtaposition of non-irrigated small parcels
of diverseannual crops, pasture and/or permanent crops
231. Pastures
2431. Land principally occupied by non-irrigated
agriculture, with significant areas of natural vegetation
2433. Land principally occupied by pastures,
with significant areas of natural vegetation
244. Agro-forestry areas (annual crops or grazing
land under the cover of forestry species)
3. Forest and semi-natural areas
correlation with nitrate was observed on large scales
(r ¼ 0.504, p < 0.001 for CD; r ¼ 0.491, p < 0.001 for DA) and for
autumnewinter samples (rainy season), a logical effect stating
that non-irrigated cropland is rain fed.
Regarding ammonium, the highest correlation values were
recorded for irrigated cropland on a large scale (DA) in
autumnewinter (r ¼ 0.510, p < 0.001). The same seasonal
pattern but with a lower correlation value was observed for
the relationship ammoniumeurban use (r ¼ 0.459, p < 0.001) at
DA scale. For non-irrigated cropland and forest, the correla-
tion values with this nutrient were lower and less significant
(Table 4).
The correlation values between phosphate and land uses
were in general the lowest of all nutrients analysed, showing
the maximum value in the case of urban use for large scales
(CD, DA) and in springesummer samples (r ¼ 0.357, p < 0.001
and r ¼ 0.348, p < 0.001 respectively).
3.2. Range of land use percentage by scale
As mentioned in Section 1.2, in a multi-scale approach
different scales would provide different land use datasets
(Fig. 3). In order to demonstrate these differences statistically,
a KruskaleWallis test was performed. Results of this test
showing that observations (% of each land use class per
 w a t e r r e s e a r c h 4 6 ( 2 0 1 2 ) 2 7 5 9 e2 7 7 1 2765

and forest (H3,516 ¼ 19.741, p < 0.001). Therefore, percentages

Table 3 e Mean values, standard deviation (SD) and range
of nutrient concentration (mg LL1) measured during obtained for each land use class (URB, IRR, NIR, AGR, FOR) were
2001e2009. AW, autumnewinter season; SS, significantly different across the four spatial scales. As
springesummer season. expected, all land uses showed a wide range of percentage
n Mean SD Minimuma Maximum values at local scales (B1 and B5) and corridors (CD) (Fig. 3)
except for urban land use, which never reached a value higher
Total N-NO 3 202 2.612 5.111 0.010 30.000
than 30% irrespective of scale. Intensive land uses near river
N-NHþ 4 213 0.166 0.515 0.010 5.500
banks such as irrigated croplands were also better represented
P-PO3
4 156 0.467 1.570 0.010 12.000
at those scales (B1, B5 and CD) (Fig. 3b). In fact, when the largest
AW N-NO 3 68 1.793 2.325 0.010 10.900 scale (DA) was selected, only the most extensive land uses
N-NHþ 4 68 0.249 0.853 0.010 5.500
(non-irrigated agriculture, total agriculture and forest) showed
P-PO3
4 68 0.149 0.324 0.010 1.380
a wide range of percentage values (Fig. 3cee, respectively). The
SS N-NO 3 134 31.000 6.009 0.010 30.000 lack of a wide range for some land uses may lead to a masking
N-NHþ 4 146 19.000 0.208 0.010 1.400
3
of those land uses which are poorly represented by those
P-PO4 88 77.000 2.038 0.010 12.000
better represented. Therefore, for the study area and the land
a Detection limit by chemical analyses. uses analysed, the largest spatial scale tested (DA) does not
seem to be the most appropriate for the study of river eutro-
scale) did not come from the same distribution were as phication due to the poor representation of irrigated agricul-
follows: total agriculture (H3,516 ¼ 9.493, p < 0.05), irrigated ture (only low percentage values). These results demonstrate
cropland (H3,516 ¼ 41.489, p < 0.0001), non-irrigated cropland the influence of spatial scale on land use quantification and
(H3,516 ¼ 32.110, p < 0.0001), urban (H3,516 ¼ 83.796, p < 0.0001) therefore on detecting river eutrophication.

Table 4 e Spearman’s rank correlation coefficients for land use classes at different spatial scales and seasons. IRR, irrigated
agriculture; NIR, non-irrigated agriculture; AGR, total agriculture; URB, urban; FOR, forest; B1, 1 km upstream radius buffer;
B5, 5 km upstream radius buffer; CD, 100 m wide upstream subcatchment corridor; DA, total drainage area of upstream
subcatchment; AW, autumnewinter; SS, springesummer. Signification level (*<0.05, **<0.01 and ***<0.001) and sample
size (n) are shown. Values in bold highlight the trend followed by the relationship nitrate-irrigated agriculture: higher
correlation values at local scales decreasing towards large scales.
B1 B5 CD DA

AW SS AW SS AW SS AW SS

N-NO
3 IRR 0.445*** 0.451*** 0.351** 0.422*** 0.266* 0.292*** 0.243* 0.315***
n ¼ 68 n ¼ 134 n ¼ 68 n ¼ 134 n ¼ 68 n ¼ 134 n ¼ 68 n ¼ 134
NIR 0.309* 0.484*** 0.319*** 0.504*** 0.433*** 0.491*** 0.449***
n ¼ 68 n ¼ 68 n ¼ 134 n ¼ 68 n ¼ 134 n ¼ 68 n ¼ 134
AGR 0.577*** 0.480*** 0.467*** 0.427*** 0.423*** 0.405*** 0.484*** 0.471***
n ¼ 68 n ¼ 134 n ¼ 68 n ¼ 134 n ¼ 68 n ¼ 134 n ¼ 68 n ¼ 134
URB 0.229** 0.349** 0.337*** 0.390*** 0.436*** 0.427***
n ¼ 134 n ¼ 68 n ¼ 134 n ¼ 134 n ¼ 68 n ¼ 134
FOR 0.550*** 0.485*** 0.470*** 0.445*** 0.339** 0.320*** 0.484*** 0.480***
n ¼ 68 n ¼ 134 n ¼ 68 n ¼ 134 n ¼ 68 n ¼ 134 n ¼ 68 n ¼ 134

N-NHþ
4 IRR 0.252* 0.290*** 0.315** 0.387*** 0.418*** 0.345*** 0.510*** 0.398***
n ¼ 68 n ¼ 146 n ¼ 68 n ¼ 146 n ¼ 68 n ¼ 146 n ¼ 68 n ¼ 146
NIR 0.177* 0.207* 0.261** 0.270* 0.288***
n ¼ 146 n ¼ 146 n ¼ 146 n ¼ 68 n ¼ 146
AGR 0.259** 0.274* 0.309*** 0.309*** 0.279* 0.341***
n ¼ 146 n ¼ 68 n ¼ 146 n ¼ 146 n ¼ 68 n ¼ 146
URB 0.261* 0.306* 0.195* 0.335** 0.279*** 0.459*** 0.188*
n ¼ 68 n ¼ 68 n ¼ 146 n ¼ 68 n ¼ 146 n ¼ 68 n ¼ 146
FOR 0.264** 0.282* 0.316*** 0.259* 0.406*** 0.297* 0.346***
n ¼ 146 n ¼ 68 n ¼ 146 n ¼ 68 n ¼ 146 n ¼ 68 n ¼ 146

P-PO3
4 IRR 0.251* 0.248* 0.270*
n ¼ 88 n ¼ 68 n ¼ 88
NIR 0.281* 0.239* 0.303* 0.268* 0.335** 0.258*
n ¼ 68 n ¼ 88 n ¼ 68 n ¼ 88 n ¼ 68 n ¼ 88
AGR 0.305** 0.297* 0.230* 0.332** 0.358** 0.289**
n ¼ 88 n ¼ 68 n ¼ 88 n ¼ 68 n ¼ 68 n ¼ 88
URB 0.241* 0.357*** 0.348***
n ¼ 68 n ¼ 88 n ¼ 88
FOR 0.304** 0.300* 0.242* 0.312** 0.355** 0.321**
n ¼ 88 n ¼ 68 n ¼ 88 n ¼ 68 n ¼ 68 n ¼ 88

81
2766 w a t e r r e s e a r c h 4 6 ( 2 0 1 2 ) 2 7 5 9 e2 7 7 1

Fig. 3 e Arrangement of sites (n [ 130) by land use percentages through the four spatial scales analysed. Land use classes:
a) urban, b) irrigated agriculture, c) non-irrigated agriculture, d) total agriculture, e) forest. Local scales: B1, 1 km radius
upstream buffer; B5, 5 km radius buffer upstream from a site. Large scales: CD, 100 m wide corridor drainage area upstream
from a site; DA, drainage area upstream from a site. Close rhombus represents the percentage of each land use class
obtained at a particular spatial scale and for each site.

3.3. Spatial autocorrelation high correlation values can be due to pseudoreplication

When sites located downstream show overlapping of drainage
areas (nested subcatchments), a problem of spatial autocor-
relation may arise. This is more probable for large drainage
basins where each site shares a great part of its drainage area
with the upstream preceding site (Fig. 2a and b). In these cases
geographical proximity of sites and the directional component
of spatial autocorrelation can lead to an absence of indepen-
dence among data (Legendre and Legendre, 1998). Thus, land
use composition obtained at each spatial scale as well as
nutrient concentration were checked for spatial autocorrela-
tion using the Mantel test. In the case of large scales, this test
found significant autocorrelation in land use composition
for total drainage area (DA: r ¼ 0.066, p < 0.05) and corridors
(CD: r ¼ 0.153, p < 0.001) while no autocorrelation was detec-
ted at local scales (buffers B1 and B5) for the response variable.
Therefore, the underlying assumption of data independence
might be violated and Spearman’s correlation coefficients
must be taken with care for large scales (CD, AD) since the
82
(Hurlbert, 1984).
3.4. Agricultural eutrophication: irrigated vs. non-
irrigated cropland
Taking into account the correlation analysis between nutrient
concentration and land use (Table 4), and the problems derived
from both the lack of a wide range of land use percentages and
spatial autocorrelation among sites, attention was focused on
nitrate level at the local scale. The criteria for making such
a choice can be outlined as follows: urban land use only
reached low percentage values and its effect could be masked
by other land uses; forest land use did not contribute to river
eutrophication; large scales were not appropriate due to
spatial autocorrelation and also due to the narrow range in
percentage values of irrigated cropland (Fig. 3); and finally,
nutrients other than nitrate showed the highest correlation
values at large scales. Thus, the impact of eutrophication due
to irrigation practices became more evident and reliable for
 w a t e r r e s e a r c h 4 6 ( 2 0 1 2 ) 2 7 5 9 e2 7 7 1
nitrate, at the most local scale of 1 km buffer (B1), and for
springesummer data (Table 4). In order to check for significant
differences in nitrate concentration between irrigated and
non-irrigated agriculture, a non-parametric test of mean
comparison (ManneWhitney) between these two groups was
performed. One group (irrigated) included sites with more than
10% of irrigated land and 0% of non-irrigated cropland. The
second group (non-irrigated cropland) included sites with
more than 10% of non-irrigated cropland and 0% of irrigated
land. In order to avoid influences other than agriculture, data
from sites impaired by high point pollution were removed. The
unilateral ManneWhitney test showed that the nitrate values
from the irrigated cropland group were significantly higher
than those from the non-irrigated group (U ¼ 48.500, p < 0.01,
a ¼ 0.05). Such an eutrophication pattern can be illustrated in
Fig. 4. Linear regression analyses showed that an increase of
land dedicated to irrigated agriculture led to an increase in
nitrate concentration (R2 ¼ 0.680, p < 0.0001, n ¼ 17). In the case
of non-irrigated cropland, the relationship with nitrate was
not significant. The performance of a breakpoint regression
analysis showed a significant threshold of about 50% (Break-
point ¼ 49.55, R2 ¼ 0.74, p < 0.01, n ¼ 17) of irrigated agriculture
(Fig. 5). Thus, the relationship between percentage of irrigated
agriculture and nitrate concentrations may be represented by
two trends: a non-significant horizontal segment for under
Fig. 4 e Irrigated vs. non-irrigated agriculture. Nitrate
concentration (NOL 3 -N mg L
L1
) against percentage of
agricultural land at the 1 km buffer scale (B1) and
springesummer season. (a) Sites with more than 10% of
irrigated agriculture and 0% of non-irrigated agriculture.
(b) Sites with more than 10% of non-irrigated agriculture
and 0% of irrigated agriculture. Regression analyses were
performed after data transformation of land use
percentages (arcsin [(3/100)1/2]) and nitrate concentration
(log( y D 1)).
83
2767
Fig. 5 e Breakpoint regression analysis of nitrate
concentration against percentage of irrigated agriculture.
Regression analyses were performed after data
transformation of land use percentages (arcsin [(3/100)1/2])
and nitrate concentration (log( y D 1)).
50% of irrigated croplands followed by a sloping line explaining
74% of the variance and described by the function
y ¼ 1.580x  0.679. These results imply a clear measure of
management: development of irrigated agriculture higher
than 50% within a 1 km radius from the river banks leads to
river eutrophication in the study area.
4. Discussion
4.1. Influence of spatial scale
One of the main goals of this study was to demonstrate that
spatial scale may affect results relating land uses to stream
eutrophication. Each particular spatial scale entails some
considerations that must be taken into account; otherwise
results may be unreliable, giving way to misleading interpre-
tations. The most important considerations are (1) the lack of
a wide range of land use percentages to be analysed and (2)
spatial autocorrelation among sites. Regarding the first issue,
a wide range of percentages for land use must be reached in
order to relate these percentages to nutrient concentration. In
this study, when land uses within the whole drainage area of
subcatchments (DA) were taken into account, percentages of
non-irrigated cropland ranged from 0 to 80%, while percent-
ages of irrigated cropland hardly reached 20% (Fig. 3b and c).
Thus, despite the fact that irrigation could be more important
in terms of eutrophication, most extended land uses may
mask the effect of irrigation cropland just because this land
use class was poorly represented. Consequently, Spearman’s
rank correlations were higher for non-irrigated than irrigated
cropland, but to conclude that non-irrigated agriculture is
more influential in eutrophication than irrigation practices
would be a misleading and unreal interpretation. Therefore,
results were scale-dependent in this study. This fact demon-
strates the need to consider multi-scale approaches for
unravelling the dynamics of water quality over space and time
(Chang, 2008).
Regarding the spatial scales assessed in the present study,
those based on 100 m wide corridors (CD) and 1 km buffers
2768 w a t e r r e s e a r c h 4 6 ( 2 0 1 2 ) 2 7 5 9 e2 7 7 1
(B1) provided the most complete range for irrigated cropland
(Fig. 3) mainly because this land use is usually located close to
river banks. However, spatial autocorrelation was detected in
data from drainage areas (DA) and corridor (CD) scales (large
scales) by means of a Mantel test. Thus, any statistical test
requiring independence of data, such as correlation or
regression analysis, would not be suitable (Legendre, 1993)
and should be applied only as exploratory analyses. In this
study, spatial autocorrelation detected on large scales (DA and
CD) was due to subcatchment overlapping which produced
pseudoreplication and enhanced correlation values. Some
researchers avoid autocorrelation and subcatchment over-
lapping by selecting single sites in independent subcatch-
ments (Lassaletta et al., 2009; Snyder et al., 2003) despite the
fact that it is difficult to reach a satisfactory sample size when
this strategy is selected. The approach suggested in this study
is to use those spatial scales which allow a satisfactory sample
size while, at the same time, providing no autocorrelated data.
A Mantel test indicated that data from 1 and 5 km radii
buffers were not autocorrelated. In addition, ranges of land
use percentages for these local scales were wider than for
large scales such as subcatchment drainage areas or sub-
catchment corridors. Therefore, both local scales can be
considered as appropriate for assessing land use effect on
river pollution by means of correlation analysis, except for
urban land use which never reached more than 28%, as
usually occurs in agricultural landscapes.
Even though the effect of urban land use on stream
eutrophication could be masked by the effects of better rep-
resented land uses, positive significant correlation values
relating urban land use to nitrogen compounds were found.
The intensity of urban influence was higher in spring-
esummer, probably due to the decrease in water flow (Robles
et al., 2002) coupled to the increase in population and waste
water effluents during summer holidays (National Statistics
Institute, www.ine.es). Correlation values relating urban use
with phosphate level at local buffer scales were not signifi-
cant, and this trend may be due to the fact that an important
part of phosphate comes from effluents of urbanized or
industrial zones (source point pollution) (Vega et al., 1998) that
are not included in local buffers. In addition, a weak positive
correlation was found between phosphate and agricultural
uses supporting the idea that a part of the phosphate comes
from the leaching of crop fertilizers. The strong relationship
between agriculture and nitrate concentration detected at 1
and 5 km buffer scales contrasted with the results of Strayer
et al. (2003) who observed a weak influence of land uses
within a 300 m radius buffer on the nitrate flux. This result
was probably obtained because the size of buffers might not
include the surrounding agricultural land uses beyond
riparian vegetation.
4.2. Agricultural eutrophication: irrigated vs. non-
irrigated cropland
Results of a correlation analysis indicated that agriculture
(AGR) measured at local scales (B1 and B5) was the most
important land use in terms of river eutrophication due to
nitrate (Spearman’s coefficient from 0.427 to 0.577 depending
on season and spatial scale, p < 0.001, n ¼ 68e134; Table 2).
84
Other studies carried out in Spain (Moreno et al., 2006;
Lassaletta et al., 2009) also relate high nitrate concentration
in streams with high percentages of agriculture in the
drainage area by means of correlation analysis and regression
(R2 ¼ 0.69, p < 0.001, n ¼ 30; r ¼ 0.632, p < 0.001, n ¼ 61, respec-
tively). Similar correlation values were obtained by Dodds and
Oakes (2008) between agriculture and nitrate when land use
was estimated for riparian corridors throughout the entire
watersheds (r ¼ 0.623, p < 0.05, n ¼ 68) and riparian corridors
adjacent to the first-order streams (0.650, p < 0.05, n ¼ 68).
Buck et al. (2004) found a strong relationship between agri-
culture and nitrate (r ¼ 0.67, p < 0.01, n ¼ 24e35) but only when
first-order streams were excluded from the analysis. Other-
wise, correlation values were not significant; probably
because the agricultural land in their study area was mainly
pastures (less intensive than agriculture in our study area)
located at the middle and lower reaches of the catchment.
Some studies do suggest, however, that urban land use is
a more important predictor of water quality than agriculture,
irrespective of the scale (Sliva and Williams, 2001; Chang,
2008). This result can be expected in highly urbanized areas
where urban surface is an important component of non-point
pollution, as in the watersheds studied by Chang (2008).
Nevertheless, Sliva and Williams (2001) point out that this
result may have been influenced by point-sources associated
with urbanized areas in their study area.
Non-irrigated agriculture is in essence not intensive due to
climatic constraints but, when irrigation is applied, farming
practices tend to intensify: water consumption of crops during
hot summers must be compensated by irrigation inputs
associated to a continued use of N fertilizers exceeding
optimum rates (Diez et al., 1997). In the study area, irrigated
lands are located close to the rivers and, as in the case of
maize, they receive a quantity of fertilizers four-fold higher
than non-irrigated cropland (Consejerı́a de Industria, Energı́a
y Medio Ambiente, 2010). This, of course, means that
nutrient inputs are higher and mobilized faster under irriga-
tion practices. When N fertilization exceeds that required for
the crop, this surplus is considered a risk for river eutrophi-
cation (Smith et al., 1999). Fertilization surpluses are often
detected throughout Spain: e.g. Isidoro et al. (2006) described
an application of 398e453 kg-N/ha/year to irrigated maize
crops located in La Violada district (NE Spain). In areas
vulnerable to nitrates located within the study area, N fertil-
ization limits have been established in 50e60 kg-N/ha/year for
most common non-irrigated cropland, 80e100 kg-N/ha/year
for irrigated cereals, 200e210 kg-N/ha/year for maize, and
80e100 kg-N/ha/year for sunflower (Consejerı́a de Industria,
Energı́a y Medio Ambiente, 2010). These facts explain the
high impact of irrigated agriculture in terms of eutrophication
when a threshold of approximately 50% of irrigated agricul-
ture is detected (Fig. 5). A threshold can be defined as the point
at which there is an abrupt change in an ecological quality,
property, or phenomenon or where small changes in a river
can produce large responses in the ecosystem (Groffman et al.,
2006). Thus, our results suggest that surpassing the threshold
of 50% of irrigated agriculture in a radius of 1 km from a river
bank would lead to a rapid and important increase in nitrate
concentration and, therefore, to river eutrophication. In other
studies, Wang et al. (1997) observed a decline in habitat quality
 w a t e r r e s e a r c h 4 6 ( 2 0 1 2 ) 2 7 5 9 e2 7 7 1
Fig. 6 e Methodological approach proposed to choose the appropriate spatial scale when studying river eutrophication
caused by agriculture.
when 50% of agriculture was exceeded in both total catch-
ment and 100-m corridors. These facts support the idea that
further research in predicting pressure thresholds is needed
due to the importance in ecosystem responses. Ecosystem
recovery by means of restoration projects might require far
more resources and time than avoiding entering the eutro-
phication state (Dodds et al., 2010).
Our results were clear and consistent: irrigation practices
are more influential in nitrate export than non-irrigated
agriculture as Scanlon et al. (2005) stated for groundwater
quality. Johnson et al. (1997) also studied two types of agri-
culture, one more intensive (rowcrop agriculture) than the
other (non-rowcrop agriculture). These authors found a strong
relationship between nitrate and rowcrop agriculture
(R2 ¼ 0.654, p < 0.0001, n ¼ 62) very similar to the relationship
between nitrate and irrigated cropland described in the
present study (R2 ¼ 0.680, p < 0.0001, n ¼ 17).
As regards seasonality, nitrate leaching in non-irrigated
croplands was associated with the rainy season, while for
irrigated lands it increased during the dry season (irrigation
season). The correlations reached between irrigated cropland
and nitrate concentration for springesummer data were
higher than for autumnewinter samples whereas, for non-
irrigated croplands, the correlations were higher in the rainy
season. This seasonal nutrient pattern has also been recorded
in northern Spanish basins (Isidoro et al., 2006) and is sup-
ported by correlation values between nitrate and total agri-
culture class (AGR): at local scales (B1 and B5) and for
85
2769
springesummer data, the relationship between nitrate and
both total agriculture and irrigation cropland was very similar.
However, for autumnewinter data, correlation values
between nitrate and total agriculture were higher than those
reached when considering only irrigated cropland. This result
could be explained by the increase in the impact of non-
irrigated cropland in the autumnewinter season.
Summarizing, the fact that reduced land use such as irri-
gated cropland caused a higher impact than more extended
land uses supports the idea that measurement of agricultural
intensity may be a better indicator of the impacts within
a river system than the percentage of differing land uses
(Harding et al., 1999). On the other hand, the variety of results
obtained among different spatial perspectives throughout the
literature suggests that different land uses may have distinc-
tive particularities and operate at different spatial scales.
Therefore, multi-scale approaches adapted to land use
particularities could be the better choice when assessing their
influence on stream water quality.
5. Conclusions
(1) Irrigated cropland was determined as the main land use
responsible for eutrophication in streams of south-central
Spanish rivers, suggesting that the intensity of agriculture
and its proximity to river banks are important factors to
take into account.
2770 w a t e r r e s e a r c h 4 6 ( 2 0 1 2 ) 2 7 5 9 e2 7 7 1
(2) A pressure threshold was found at 50% of irrigated agri-
culture within a 1 km radius of river banks. Surpassing this
percentage value may lead to a rapid increase in nitrate
concentration in streams and, therefore, to river
eutrophication.
(3) The election of the spatial scale may have important
influences on the final results and ecological interpretation
of studies assessing the relationship between land uses
and eutrophication in streams.
(4) In the study area, the adequate spatial scale to assess the
impact of non-irrigated and irrigated agriculture in stream
water quality was a 1 km radius buffer upstream from sites.
(5) Depending on the subject under study, the most appro-
priate spatial scale may vary and it may even be possible to
find more than one adequate scale. As a new contribution,
the following methodological approach may be outlined
when analysing relations between land use and eutro-
phication indicators (Fig. 6):
a) It is important to check for the spatial scale(s) that
provide(s) more complete percentage ranges for all
land uses under study. The wider the range of land use
percentages analysed, the more reliable the results
obtained.
b) Secondly, it is also necessary to check for autocorre-
lation of data when geographical distance between
sites is short (a few kilometres, sites located within the
same reach or segment), and always in the case of
nested subcatchments.
c) The most appropriate spatial scale(s) to relate land
uses to river eutrophication will be the one that
provides the largest number of sites without gener-
ating data autocorrelation.
This work suggests that a multiple perspective may
provide valuable information when relating land use to
stream eutrophication. This multiple perspective refers not
only to the spatial scale but also the segregation of land use
classes. In further research, more precise distinctions
between land uses that are pooled into one land use class
(agriculture in general), together with more information
about pressure thresholds, would provide interesting infor-
mation about the effect of different types of agricultural
practices such as irrigated cropland, organic farming or
precision agriculture, among others. Finally, as a main
precept for river management, irrigated cropland should not
be located close to river banks (<1 km) and, if allowed,
should generally not surpass the 50% threshold of
irrigated vcroplands around rivers. In addition the develop-
ment of good agricultural practices in terms of avoiding
surpluses in fertilization would also help to prevent stream
eutrophication.
Acknowledgements
We would like to thank all reviewers for their helpful
comments, which improved the quality and understanding of
this paper. Special thanks to Walter K. Dodds for his sugges-
tions and to Kathryn Ann Walsh for the review of English
language. This study was financed by the Regional Project PAI-
86
06-0057 granted by the Junta de Comunidades de Castilla-La
Mancha.
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88
5.5. ¿Son las cianobacterias bentónicas indicadores de presiones
antropogénicas en los sistemas fluviales?

Título original: Could benthic freshwaters Cyanobacteria be biological

indicators of anthropogenic pressures?

Autores: Laura Monteagudo y José Luis Moreno

Publicación: Under Review en Ecological Indicators
Impact Factor: 3.444
Fecha 2015

RESUMEN

El uso de las cianobacterias como indicadores biológicos de presiones

antropogénicas y en la evaluación del estado ecológico de las masas de agua es
bastante limitado en comparación con otros elementos biológicos. Esto puede
deberse fundamentalmente a dos razones: (1) el conocimiento limitado sobre su
respuesta a estas presiones; (2) la complejidad en la identificación de ciertas
especies de cianobacterias para técnicos e investigadores. Sin embargo, las
cianobacterias, como otras macroalgas, son organismos estrechamente ligados a las
condiciones del agua y pueden ser potenciales indicadores de las presiones
humanas, pudiendo llegar a reforzar los índices basados en macrófitos.

En este trabajo se analizó la variación en la comunidad de cianobacterias como

respuesta a ocho variables ambientales (pH, conductividad, temperatura, altitud,
nitrato, ortofosfato y dos usos de suelo: regadío y secano) en 85 estaciones de
muestreo a lo largo de la región. Con ello, se evaluó la capacidad de las
cianobacterias como indicadores de dos factores de estrés: el fosfato procedente de
las aguas residuales urbanas y el nitrato derivado de los terrenos agrícolas.

Los análisis de modelos lineales realizados (Distance-based linear modelling)

mostraron que las variables que mejor explicaban la variación en la composición de

89
especies fueron el pH y la conductividad. Además, se observaron diferencias entre la
especies con y sin heterocitos. Mediante la técnica del ‘linkage tree’ o árbol de
agrupación, se dividió la muestra en grupos biológicos diferentes atendiendo a
diferencias en las variables ambientales por orden de influencia. Esto reveló que la
variable ambiental más influyente en la variedad biológica entre grupos fue la
conductividad. Comparando los nutrientes, el ortofosfato fue más determinante para
la composición de especies que el nitrato. El análisis de porcentajes de similaridad
mostró los porcentajes de contribución de cada especie a la similaridad total de cada
grupo. Así, Coleodesmium wrangelii, Nostoc caeruleum y Phormidium fonticola
resultaron ser las especies principales en los sitios con valores más bajos de
conductividad (<30µS·cm-1), sin presiones antropogénicas. P. favosum fue la especie
más común en los sitios con los valores más altos de ortofosfato (>0.15 mg P-PO 4 3-·l-
1), es decir, aquellos afectados por vertidos urbanos. Por otro lado, en ambientes con
valores bajos de ortofosfato (<0.02 mg P-PO 4 3-·l-1) las especies del género Rivularia
fueron las más comunes. Al contrario de lo esperado, una cianobacteria con
heterocitos fue la más frecuente en sitios con los valores más elevados de nitrato (>2
mg N-NO 3 -·l-1), independientemente de si el origen de este compuesto era natural o
procedente de la agricultura. En ambientes bajo presión agrícola, caracterizados con
valores elevados de nitrato y con un alto porcentaje de regadío en su proximidad,
una de las especies más comunes fue P. autumnale. Por último, N. verrucosum mostró
una gran tolerancia a la mayoría de las variables ambientales medidas, exceptuando
las derivadas de las presiones humanas.

De todos estos resultados se puede deducir que algunas cianobacterias pueden

ser útiles en la bioindicación de las presiones antropogénicas (p. ej. Nostoc
verrucosum, Phormidium autumnale, Plectonema tomasinianum, Rivularia
haematites, Tolypothrix distorta); mientras que otras ofrecen más información
acerca de las condiciones físico-químicas naturales del agua en sitios no
afectados por las actividades humanas (p. ej. N. caureuleum, Phormidiumm
fonticola). Por tanto, ampliar el conocimiento sobre la relación entre
comunidades de cianobacterias y otras macroalgas con el medio ambiente,
en diferentes escenarios de impacto, podría ayudar a mejorar la precisión de los
índices basados en macrófitos en un futuro.

90
Could benthic freshwater Cyanobacteria be biological indicators of
anthropogenic pressures?
Laura Monteagudo* and José Luis Moreno
University of Castilla-La Mancha, Regional Center of Water Research (CREA)
Crta. de Las Peñas km. 3, Albacete 02071, Spain.
*Corresponding author: Tel.: +34 967599200x2576;fax:+34 967599269; E-mail: laura.monteagudo.c@gmail.com

ABSTRACT
Cyanobacteria are hardly used as biological indicators of anthropogenic pressures or for assessing the ecological
status of rivers, possibly for two main reasons: a) their response to anthropogenic pressures is often poorly
known; b) reliably identifying cyanobacteria species is a challenge for technicians and researchers. However,
some species can be good indicators of anthropogenic pressures and could, therefore, reinforce macrophyte
methods to assess ecological status of rivers. We analysed variation in benthic cyanobacterial assemblages at 85
sites in (mainly agricultural) South-Central Spain as a response to eight environmental variables: pH,
conductivity, temperature, altitude, nitrate, orthophosphate, irrigation land use and non-irrigation land use. We
assessed the usefulness of cyanobacteria species as biological indicators of two human stressors: the high
orthophosphate concentration that derives from urban wastewaters; the high nitrate concentration produced by
agricultural land use.
Distance‐based linear modelling showed that pH and conductivity defined the best model to explain variation in
species composition. Differences in the response of heterocystous and non-heterocystous cyanobacteria were
found. A linkage tree was used to generate divisive biotic samples clustering according to significant differences
in environmental variables. Conductivity was the main environmental factor that contributed to differences
between assemblages. Othophosphate was a more influential nutrient for community composition than nitrate. A
similarity percentage analysis revealed that Coleodesmium wrangelii, Nostoc caeruleum and Phormidium
fonticola were the main contributors at very low conductivity sites (30 µS·cm-1) with no human pressures. P.
favosum was the commonest species at the sites with the highest orthophosphate levels (>0.15 mg P-PO 4 3-· l-1)
impacted by urban wastewater. Rivularia species were dominant at the sites characterised by low
orthophosphate values (<0.02 mg P-PO 4 3-· l-1), but not impacted by urban wastewater. Unexpectedly, a
heterocystous cyanobacterium (R. haematites) became the most frequent species at the sites with highest nitrate
values (> 2 mg N-NO 3 -·l-1), regardless of being impacted by agriculture or not. P. aumtumnale was present at the
impacted sites with the highest orthophosphate and irrigated agriculture values. Finally, Nostoc verrucosum
tolerated a wide range of environmental conditions, but avoided anthropogenic pressures, and was absent at the
sites with high orthophosphate, nitrate or irrigation land use values.
These results suggest that some cyanobacteria species (e.g. Nostoc verrucosum, Phormidium autumnale,
Plectonema tomasinianum, Rivularia haematites, Tolypothrix distorta) could be useful tools for the bioindication
of anthropogenic pressures, while others provide more information about natural physicochemical reference
conditions (N. caeruleum, Phormidium fonticola). Further research into cyanobacteria and macroalgae
assemblages in different impacted scenarios could help improve macrophyte indices.
1. INTRODUCTION those based on macroinvertebrates or diatoms
(Ferreira et al., 2005). Demars et al. (2012)
Since the Water Framework Directive (WFD)
reviewed several macrophyte indices and
came into force, EU Member States are
highlighted the poor accuracy observed in this
required to achieve a ‘good status’ in all surface
metrics when applied to different regions.
and groundwater bodies, which implies
Some criticisms about macrophyte indices
achieving a ‘good chemical’ and ‘good
indicate that they are based on riverine plants
ecological’ status. Regarding ecological status,
prior to exclusive aquatic taxa and the usual
Member States shall establish monitoring
exclusion of benthic macroalgae (including
systems to estimate the values of the biological
cyanobacteria), despite these organisms being
quality elements specified per surface water
closely linked to water properties. Such
category (WFD - Annexe V - 1.4.1.). This fact
discrimination could be due to a poor
has led to increasingly used metrics based on
understanding of their response to
different groups of organisms (e.g.
anthropogenic pressures (Thacker and Paul,
invertebrates, diatoms or macrophytes) to
2001), and technicians and researchers’
assess ecological status. In general, indices
difficulty in reliably identifying several species
based on macrophytes are less popular than
(Marquardt and Palinska, 2007). Despite these

91
difficulties, studies worldwide have related
cyanobacteria to several human pressures
responsible for eutrophication in freshwater
ecosystems, such as agricultural loadings,
urban pollution or industrial discharges (e.g.
Johansson, 1982; Jafari and Gunale, 2006;
Parikh et al., 2006). Although cyanobacteria
have been usually included in studies that have
related algal communities to environmental
factors (e.g. Potapova et al., 2005; Dell’Uomo
and Torrisi, 2009), some physiological features
have established remarkable differences in the
way they are influenced by nutrients. Unlike
other algae, cyanobacteria are able to fix
atmospheric nitrogen, which allows them to
grow at low rates of dissolved nitrogen
compounds. Some authors have suggested that
this characteristic may diminish their
dependence on nitrogen availability in the
water column (e.g. Larkum et al., 1988). In
addition, heterocysts, allow some
cyanobacteria to fix atmospheric nitrogen (N 2 )
under aerobic conditions, while N 2 fixation is
limited to anaerobic and (or) dark conditions
with non-heterocystous cyanobacteria (Potts,
1979; Lee, 2008). Loza et al. (2014) suggested
that this ecophysiological advantage could
explain not only the dominance of
heterocystous species at low levels of
combined nitrogen, but also the preference of
non-heterocystous species for high levels of
these compounds. With phosphorus, certain
cyanobacteria, such as Rivularia sp., are able to
live at phosphorus limitation because of
phosphatase activity, which is a good indicator
of oligotrophic conditions (Mateo et al., 2010).
All these facts evidence that cyanobacteria are
a large diverse group of organisms which
includes freshwater species that grow at
unimpaired and impaired sites. Thus some of
these species are expected to be suitable
indicators to account for the development of
biological indices of water quality. In order to
identify them, it is essential to examine the
relationship between the cyanobacteria
community and the surrounding environment
conditions in depth, and to also apply
requirements for good indicator species. Hence
the main objectives of this study were to: (1)
analyse which variables determine the benthic
cyanobacteria community in the study area; (2)
examine the differences between
92
heterocystous and non-heterocystous species;
(3) identify which species of cyanobacteria in
the study area are suitable as indicators of
human pressures.
2. MATERIAL AND METHODS
2.1. Study area and sampling design
This study was carried out in Castilla-La
Mancha (South-Central Spain), a region marked
by agriculture as its main anthropogenic land
use. As semi-arid climatic conditions
predominate in the study area, irrigation is
needed for high-productivity crops. Thus
6.26% of agriculture is conducted under
irrigation conditions (Spanish Survey of
Surfaces and Crop Yields (ESYRCE), 2013), and
these cultures are usually located on the banks
of streams and rivers.
Sites were selected to cover the most complete
gradient of environmental conditions, such as
lithology and altitude, among others (Figure 1).
Regarding lithology, three main types were
defined: calcareous (Cretacic and Jurassic
limestones), mixed (Quaternary sedimentary
valleys) and siliceous (granites, slates and
shales). Reaches were waded in an upstream
zigzag pattern and sampling was addressed in
all habitat types. Thus the macroscopic
cyanobacteria thalli that grew submerged or in
the splash zone were collected by hand and
fixed with 4% formalin. Species determination
was carried out in the laboratory under a Leica
M165C stereoscope and a light microscope
Olympus BX50. Glycerine-gelatine was used to
make permanent slides. In this study, we
considered only those species that
predominated in macroscopic colonies.
During sampling, pH and conductivity were
measured in situ with the appropriate sensors
(Multiline P4 WTW). Phosphate (P-PO 4 3−),
nitrate (N-NO 3 -) and ammonium (N-NH 4 +)
were determined photometrically with MERCK
kits (Spectroquant®) in the laboratory. The
percentage of four land uses (irrigated
agriculture, non-irrigated agriculture, urban
and forestry) was quantified within a 1-
kilometre radius buffer upstream of each site,
according to Monteagudo et al., 2012.
Geographic data management was carried out
with ArcGIS 9.2.
Figure 1. Study area showing land uses and location of sites. Site symbols correspond to three lithology categories:
calcareous (circles), mixed (squares) and siliceous (triangles).

2.2. Statistical analyses the ‘Best’ procedure for the variables selection
and the ‘AIC’ (“An Information Criterion”;
Boxplots of the environmental variables for
Akaike, 1973) criterion for model comparisons
each species were performed with Statistica
(Anderson et al., 2008). The criterion comes
v.9. In order to explore the relationship
from the likelihood theory and smaller AIC
between the environmental variables and the
values indicate a better model. Unlike the R2
cyanobacteria community, we performed the
criterion, AIC is not influenced by the number
DISTLM and LINKTREE routines implemented
of predictor variables (Anderson et al., 2008).
in Primer v6. Biological matrices were built
Pseudo-F is the statistic for testing the general
using the presence/absence data to avoid the
null hypothesis of no relationship, which is an
influence of factors such as hydrologic stability,
analogue of Fisher’s F ratio used in traditional
water clarity, grazers or light availability
regression. In addition to overall species
(Porter et al., 2008).
composition, DISTLM was also performed for
In order to analyse the relationship between both heterocystous and non-heterocystous
cyanobacteria assemblage and the cyanobacteria separately as they were
environmental variables, we used the distance- expected to show a different relation with
based linear model routine (DISTLM). This nitrate.
analysis performs ‘marginal tests’ for each
For a further understanding as to how the
single variable, and also tests all the possible
cyanobacteria community changes in different
combinations of variables to find the ‘best
environmental scenarios, we used the linkage
overall solution’; in other words, the best
tree analysis (LINKTREE) included in Primer
model to explain variation in biological data.
v6. This classification analysis first identifies
Among the model-building options, we selected

93
the best subset of environmental variables so
that the Euclidean distances of the scaled
environmental variables show a maximum
correlation with community dissimilarities
(biological Bray-Curtis similarity matrix); then
it uses these variables to describe how the best
assemblage samples are split into groups
(Clarke and Gorley, 2001). Thus while DISTLM
indicates which variables explain variation in
the overall species composition, LINKTREE
combines the most influencing variables,
together with the biological assemblage
similarities among sites, and successively
divides all the samples into (statistically
different) groups. The B% value obtained with
this analysis is an absolute measure of group
differences. Therefore, low B% values
correspond to the samples that come close
together in a multidimensional scaling plot. As
divisions are carried out in an influential order
of environmental variables, dissimilarity
between groups decreases in successive
divisions, as does the B% value. The
significance of difference between groups is
measured by a simultaneous SIMPROF test
(Clarke & Gorley, 2006). This routine performs
a series of “similarity profile” permutation tests
to check if each split produces groups of sites
that belong to the same assemblage or not (π
statistic). For the purpose of interpreting
biological differences between groups, we also
applied a similarity percentage analysis
(SIMPER) to identify the indicator species
within each group by calculating its
contribution based on similarity percentages.
Table 1. The species considered in this study and their frequency of appearance as number of sites (n).
Taxon
Coleodesmium wrangelii Borzì ex Geitler
Microcoleus subtorulosus Gomont ex Gomont
Microcoleus vaginatus Gomont ex Gomont
Nostoc caeruleum Lyngbye ex Bornet & Flahault
Nostoc sphaericum Vaucher ex Bornet & Flahault
Nostoc verrucosum Vaucher ex Bornet & Flahault
Phormidium autumnale Gomont
Phormidium favosum Gomont
Phormidium fonticola Kützing
Phormidium retzii Kützing ex Gomont
Phormidium uncinatum Gomont ex Gomont
Plectonema tomasinianum Bornet ex Gomont
Rivularia biasolettiana Meneghini ex Bornet & Flahault
Rivularia haematites C.Agardh ex Bornet & Flahault
Scytonema hofmannii C.Agardh ex Bornet & Flahault
Scytonema myochrous C.Agardh ex Bornet & Flahault
Tolypothrix distorta Kützing ex Bornet & Flahault
Tolypothrix robusta N.L.Gardner
3. RESULTS
3.1. Recorded species
In this study, we considered only the benthic
species that formed macroscopic thalli,
recorded at two sites, at least, with available
environmental data. Eighteen cyanobacteria
species that belong to orders Nostocales and
Oscillatoriales were selected, including five
heterocystous genera (Coleodesmium Borzì ex
Geitler, Nostoc Vaucher ex Bornet & Flahault,
Rivularia C.Agardh ex Bornet & Flahault,
Scytonema C.Agardh ex Bornet & Flahault and
Tolypothrix Kützing ex Bornet & Flahault) and
four non-heterocystous genera (Microcoleus
Desmazières ex Gomont, Phormidium Kützing
ex Gomont, Plectonema Thuret ex Gomont and
Schizothrix Kützing ex Gomont) (Table 1). The
most frequent species were Nostoc verrucosum,
Rivularia haematites and Phormidium favosum.
On the contrary, Microcoleus subtorulosus, M.
vaginatus, Phormidium uncinatum and
Tolypothrix robusta were recorded only twice.
Certain genera (e.g. Leptolyngbya sp.,
Geitlerinema sp., Schizothrix sp. or Lyngbya sp.)
were identified as secondary/occasional taxa in
macroscopic colonies, which were not taken
into account in this study. Some taxa that
formed macroscopic thalli, like
Cylindrospermum sp., Anabaena sp. or
Nostochopsis lobata Wood ex Bornet et Flahault
(Moreno et al., 2012), were also recorded in the
study area, but were excluded from this study
due to insufficient data.
n Sites
4 4 70 72 74
2 71 77
2 69 79
5 4 70 72 73 76
7 2 22 26 33 50 61 62
26 1 4 15 18 21 24 28 31 32 35 36 37 38 42 44 45 52 57 60 64 65 66 77 80 81 85
10 5 6 22 24 35 43 48 51 68 79
23 1 2 7 8 9 10 13 14 15 17 19 25 26 27 32 42 44 46 49 50 56 78 79
5 64 71 76 84 85
6 5 11 20 34 59 64
2 3 60
13 5 6 12 25 32 35 40 42 44 48 53 57 83
16 5 13 16 29 33 44 47 55 56 57 60 63 67 69 75 80
25 5 8 12 13 15 18 23 30 39 40 41 42 43 46 52 54 57 58 59 60 61 69 81 82 83
3 8 12 23
4 4 39 57 67
15 5 12 24 25 26 33 41 43 47 54 56 57 60 64 76
2 53 60
94
Figure 2. Boxplots of the environmental variables for the species considered herein. Species codes in
Table 1.

95
3.2. Environmental ranges vaginatus and both Rivularia grew at the sites
with the highest non-irrigated land use values.
The environmental ranges for each species are
compiled in Figure 2. In pH terms, the most 3.3. Distance-based linear model
tolerant cyanobacterium was N. verrucosum
Regarding species composition (Table 2.a.),
(Figure 2.a.), which was able to grow from 6.65
marginal tests revealed that pH, conductivity,
to 9.40 pH. Almost all the species were more
altitude, nitrate and non-irrigated land use
frequent at alkaline sites with pH > 8. However,
were all statistically significant variables to
Coleodesmium wrangelii, Microcoleus
explain variation in species composition when
subtorulosus and Nostoc caeruleum were
considered alone (p≤0.05). Conductivity was
stenoic species that were recorded mainly at
the most explanatory variable, despite the
acid-neutral values (pH < 7.5). According to
proportion of variation explained being low
Figure 2.b., C. wrangelii and N. caeruleum were
(5.6%). The best model was achieved by a
also recorded at the lowest conductivity levels,
combination of ‘pH’ and ‘conductivity’, which
together with Phormidium fonticola. All three
accounted for 9.3% of variability of the
are characteristic species of siliceous mountain
biological data cloud.
streams. Overall, more than 75% of the species
records corresponded to sites with
Table 2. The distance-based linear model analysis results. P, p-
conductivity values below 900 µS·cm . The -1
value; Prop. and R2, proportion of explained variation in the
wider conductivity range was shown by biological data for each variable and for the model, respectively.
Tolypothrix distorta and three Phormidium a) Species Composition
Marginal tests: Pseudo-F P Prop.
species (P. retzii, P. autumnale and P. favosum). pH 3.248 0.0004 0.038
Regarding temperature (Figure 2.c.), all the Temperature 1.164 0.3044 0.014
Conductivity 4.907 0.0001 0.056
species were found to be more frequent at 15- P-PO43- 1.285 0.1901 0.015
20ºC, and P. fonticola and N. caeruleum N-NO3
-
3.618 0.0002 0.042
preferred the coldest streams. On the contrary, Altitude 2.385 0.0123 0.028
Irrigated land use 1.566 0.1131 0.019
N.verrucosum and P. favosum grew at the Non-irrigated land use 2.977 0.0018 0.035
warmest sites and also showed the widest Best overall solution: AIC R2
temperature range. Most species were more pH & Conductivity 703.96 0.093

frequent in medium-altitude mountain streams b) Heterocystous species composition

(from 400 to 1,000 m a.s.l.), while C. wrangelii, Marginal tests: Pseudo-F P Prop.
pH 8.401 0.0002 0.092
N. caeruleum and P. fonticola were recorded Temperature 0.601 0.6012 0.007
exclusively at sites above 800 m a.s.l., which Conductivity
3-
2.690 0.0654 0.031
P-PO4 0.687 0.4896 0.008
constituted the typical assemblage in high N-NO3
-
1.972 0.1470 0.023
mountain siliceous streams (>1,000 m altitude) Altitude 0.792 0.5054 0.009
(Figure 2.d.). As shown in Figure 2.e., all the Irrigated land use 1.078 0.3964 0.013
Non-irrigated land use 2.770 0.0567 0.032
species were tolerant to low nitrate Best overall solution: AIC R
2

concentrations. However, only N. verrucosum pH & Non-irrigated agriculture 473.07 0.124

and P. favosum were found to be growing over c) Non-Heterocystous species composition

12 mgN-NO 3 -1·l-1. P. fonticola was recorded at Marginal tests: Pseudo-F P Prop.
pH 1.059 0.3770 0.013
the lowest nitrate values and showed less Temperature 1.718 0.2090 0.028
tolerance to this nutrient. Once again (Figure Conductivity 1.183 0.0001 0.125
2.f.), all the species were more frequent at low P-PO43- 1.910 0.1649 0.022
-
N-NO3 2.312 0.1053 0.027
phosphate concentration levels; indeed P. Altitude 2.683 0.0745 0.031
favosum appeared to be the most phosphate- Irrigated land use 8.788 0.0002 0.096
Non-irrigated land use 1.957 0.1602 0.023
tolerant species. In general terms, all the Best overall solution: AIC R2
species grew at sites with low agricultural land Conductivity, P-PO4
3-
& Irrigated land use 434.88 0.207
use percentages (from 0% to 20%), as shown
in Figure 2.g. and 2.h. Tolerance to agricultural With heterocystous species (Table 2.b.), pH
land use varied according to species and was the only statistically significant variable in
agriculture type. For irrigated agriculture, the marginal tests and explained 9.2% of variation,
most tolerant species were M. vaginatus, P. while the best model was that built with the pH
autumnale and P. favosum, which were able to and non-irrigated land use combination, which
grow in streams surrounded by more than accounted for 12.4% of variation. The best
80% of land used for irrigated agriculture; M. model for the non-heterocystous species
explained 20.7% of variation, and was built

96
upon the conductivity, phosphate and irrigated
land use combination.
3.4. Linkage tree and similarity
percentage contribution
By starting with the whole group of samples,
the first split (1) of the classification
dendrogram provided by LINKTREE (Figure 3)
resulted in two groups based on conductivity
values: group A, included very low conductivity
sites (<30 µS·cm-1) and was separated from the
other sites. Therefore, conductivity was the
most influential variable, as suggested by the
DISTLM results for species composition (Table
2.a.). The indicator species of group A were
Coleodesmium wrangelii, Nostoc caeruleum and
Phormidium fonticola, and they all equally
contributed (33.33%) to average within-group
similarity. Conductivity also determined splits
2 and 5 by generating groups B and E,
respectively. Group B was characterised by low
conductivity values (from 43 to 142 µS·cm-1),
while group E included sites with medium
conductivity values (Figure 2.b.). The main
species to contribute to both groups (B and E)
was N. verrucosum. The groups generated after
group E (F to J) were characterised by medium-
high conductivity values (>435 µS·cm-1). After
conductivity, the next influential variable was
temperature. This variable determined splits 3
and 7, which brought about groups C and G,
characterised by warm water sites (>20 ºC).
Therefore, the species that typified these
groups were those that tolerated higher than
average temperatures, mainly N. verrucosum,
Phormidium favosum, Rivularia biasolettiana
and Tolypothrix distorta (Figure 2.c.).
The first nutrient in order of importance was
phosphate, which determined divisions 4 and
6. The sites that were impacted by urban
wastewater gave the highest phosphate
concentration (>0.15 mg P-PO 4 3-·l-1) and were
included in group D, where P. favosum
contributed to the highest similarity
percentage. It is noteworthy that the indicator
species in group D were all non-heterocystous
species, which could explain why the best
model explained why the non-heterocystous
species composition included phosphate
among the selected variables (Table 2.c.). Split
6 separated the low from the medium
phosphate sites. Irrigation and nitrate had an
influence only in the last classification steps
once conductivity was medium-high and the
phosphate concentration was almost negligible.
97
Group H included the sites impacted by
intensive agriculture practices under irrigation,
while groups I and J included the sites with a
nitrate concentration below 1.9 mg N-NO 3 -·l-1
and over 2.0 mg N-NO 3 -·l-1, respectively. R.
haematites was the best indicator species in
these three groups, although it reached the
highest overall contribution percentage
(78.62%) in group J – a high nitrate
concentration.
4. DISCUSSION
4.1. Variables that influence the benthic
cyanobacteria assemblages in the study
area
According to the results of this study,
conductivity was the most influential variable
in the cyanobacteria species composition in the
study area, and it was only when conductivity
was not that relevant that the influence of
nutrients (phosphate and nitrate, in order of
importance) became evident. Biggs (1990) also
described conductivity as the most important
variable for distinguishing periphyton
community types, and pointed out that
conductivity integrates several higher-level
processes. Therefore, conductivity may mask
the influence of nutrients among other
variables in the cyanobacteria species
composition. For instance, the result of the
distance-based linear models on the overall
species composition indicated that the pH and
conductivity combination was the best model
to explain variation. Despite nitrate being
statistically significant in the marginal tests,
both nutrients were excluded from this model.
According to this result, community
composition would not be explained firstly by
nutrient concentration, as other studies have
reported (e.g. Larkum et al., 1988; Thacker and
Paul, 2001). However, the LINKTREE routine
helped reveal the underlying relationships
between the cyanobacteria community and
environmental variables, which are
summarised in Figure 4. Three main
conductivity groups were distinguished: very
low (<145 µS·cm-1), low (from 145 to 430
µS·cm-1) and medium-high (>430 µS·cm-1).The
very low-conductivity sites corresponded to
the unimpaired freshwater streams located on
siliceous mountains (impaired sites are
expected to show higher conductivity values).
Here the typical reference cyanobacteria
community included Nostoc caeruleum,
Coleodesmium wrangelii and Phormidium
Figure 3. Biotic clustering based on environmental variables and species contributions to groups. COND,
conductivity (µS·cm-1); T, temperature (ºC); IRR, irrigation land use (%); P-PO 4 3-, phosphate concentration
(mg P-PO 4 3-·l-1); N-NO 3 -, nitrate concentration (mg N-NO 3 -·l-1); π: SIMPROF statistic; p: p-value of the SIMPROF
test. Species codes in Table 1.

98
fonticola. This is in accordance with Kastovsky cyanobacteria considered herein. These
et al. (2010), who reported these three species conductivity levels were recorded in middle
in oligotrophic mountain waters. N. verrucosum reaches, as well as in calcareous mountains,
was also a typical species at such sites despite and regardless of them being impacted or not.
it showing better tolerance to conductivity in Under such conditions, the influence of
the study area (Figure 2). variables other than conductivity arose. Thus
the next three groups were described
The sites included in the low-conductivity
according to phosphate concentration (high-
group were located at middle reaches, but had
urban, medium and low) (Figure 4). The high-
no remarkable anthropogenic influence. The
urban-phosphate group (>0.15 mg P-PO 4 3-·l-1)
community at these sites was dominated by N.
included the sites located downstream of the
verrucosum, R. biasolettiana and Tolypothrix
urbanised areas and small villages with poor
distorta. Despite these species having been
water treatment. The typical species in this
often recorded in alkaline and mineralised
group were mainly P. favosum, followed by P.
waters (e.g. Aboal, 1988; Margalef, 1955), they
autumnale and Plectonema tomasinianium.
were found more frequently at the sites with
Both Phormidium species have been previously
lower conductivity values than the majority of
described to be sewage- and organic pollution-
species studied herein. Medium high-
tolerant (Lukavský et al., 2006 and Barinova et
conductivity values (>430 µS·cm-1) were the
al., 2010, respectively).
preferred range for the majority of

Figure 4. Summary scheme of the typical cyanobacteria assemblages according to the most relevant
variables.

The medium-phosphate group was verrucosum replaced P. autumnale when

characterised by phosphate values within the compared with the high-phosphate group
0.02-0.15 P-PO 4 3-·l-1 range, which were (Figure 4). This fact supports the statement
adequate nutrient conditions for almost all the that N. verrucosum is found mainly in habitats
cyanobacteria species compiled herein. Here N. with a low nutrient content (Mollenhauer et al.,

99
1999). Finally, the low-phosphate group
included a wide variety of sites with null or
negligible values (<0.02 mg P-PO 4 3-· l-1). When
the phosphate concentration was low, nitrate
influenced the cyanobacteria assemblages. As
expected in nitrogen-fixing taxa, phosphate had
a stronger impact than nitrate on community
composition since the influence of nitrate was
perceptible only under low phosphate
conditions. The dominant species at the low-
phosphate sites was Rivularia haematites,
irrespectively of nitrate concentration. This
could be explained by phosphatase activity,
which would allow Rivularia to metabolise
organic phosphorus and to tolerate
oligotrophic conditions (Sabater et al., 2000;
Mateo et al., 2010). Differences in community
composition, depending on nitrate
concentrations, were given by species other
than R. haematites, including heterocystous and
non-heterocystous species (Figure 4). The high
agricultural-nitrate group included the sites
located in intensive agricultural areas under
irrigation systems, which is considered the
most important origin of nitrate pollution in
the study area (Monteagudo et al., 2012). A
direct estimation of nitrate from a water
sample would provide a particular measure
that would correspond to a given time. Unlike
nitrate concentration, the percentage of land
used for irrigated agriculture would represent
an indirect long-term nitrate measure, and
would more accurately reflect the real impact
scenario. Therefore, the typical species in this
group are expected to be either nitrophilous-
or nitrate-tolerant. The characteristic
cyanobacteria assemblages in nitrogen-rich
sites influenced by irrigated agriculture
included R. haematites, P. autumnale and T.
distorta. This result agrees with Komárek and
Anagnostidis (2005), who described P.
autumnale to be a nitrophilous species.
However, the same authors related T. distorta
with unpolluted waters, which suggests that T.
distorta shows good tolerance and is able to
grow in the absence and presence of nitrate, as
indicated by Ogawa and Carr (1969). In the
medium-nitrate group, the concentration was
over 2.0 mg N-NO 3 -·l-1, and the source of this
nutrient could be anthropogenic or natural, or
even both. The sites in this group would be
located mainly on calcareous mountains with
natural nitrate inputs or in non-irrigated
agricultural areas where low fertilisation is
applied. The cyanobacteria assemblage in this
group was also dominated by R. haematites,
100
while P. favosum replaced P.autumnale. Finally,
the low-nitrate group corresponded to the
oligotrophic sites represented mainly by R.
haematites, N. verrucosum and, to a lesser
extent, by P. favosum, according to the
percentage contribution values shown in
Figure 3 (Group I). This suggests that
heterocystous species may prevail over non-
heterocystous species under low nitrate
conditions, as indicated for other regions (e.g.
Stancheva et al., 2013).
4.2. Suitability of species as indicators of
human pressures
According to Bellinger and Sigee (2010), given
their presence, good indicator species should
provide information on the surrounding
physical and/or chemical environment at a
particular site, and are expected to generally
have a narrow ecological range, a wide
geographical distribution and reliable
identification. Following these criteria, not all
cyanobacteria species are suitable as
bioindicators of the human pressures in the
study area. For instance, Nostoc caeruleum,
Coleodesmium wrangelli and Phormidium
fonticola only provided information on
conductivity conditions; Rivularia biasolettiana
appeared to be more related to temperature
than to other variables; P. favosum was seen to
be tolerant to a wide range of nitrate and
phosphate concentrations; several species
were infrequent in the study area. Conversely,
other species met the above-mentioned
requirements and could be useful tools to
develop biological indices, these being R.
haematites, N. verrucosum, P. autumnale,
Tolypothrix distorta and Plectonema
tomasinianum.
N. verrucosum is a well-known worldwide
distributed species that is normally related to
low nutrient contents (Ennis, 1978;
Mollenhauer et al., 1999). In our study area it
was the most frequent species located at the
sites characterised by low phosphate, nitrate
and agriculture land use values. Thus the
results suggest that N. verrucosum is a sensitive
species which could be a useful indicator of
absence of anthropogenic pressures.
P. autumnale exclusively characterised sites
under anthropogenic pressure, which agrees
with other authors who have reported this
species in polluted waters (Barinova et al.,
2010; Loza et al., 2013). Therefore, the
presence of this species could provide valuable
information about the water quality at a given
site, providing it is not confused with P.
favosum, a morphologically similar species that
is sometimes misidentified as P.autumnale, and
vice versa (Komárek and Anagnostidis, 2005).
P. tomasinianum has often been related to
oligotrophic conditions (Wehr and Sheath,
2003; Kastovsky et al., 2010), and less
frequently to medium to high phosphorus
concentrations (Schneider and Melzer, 2003).
In this study, P. tomasinianum appeared among
the typical species in the cyanobacterial
community of both high and medium
phosphate concentrations, which suggests its
potential as an indicator of pollution in the
study area. This divergence of results may
suggest that the suitability of some species as
bioindicators may vary in different geographic
regions.
R. haematites grew mainly at the sites with
nitrate enrichment from both agricultural and
natural sources, provided that the phosphate
concentration was low (<0.02 mg P-PO 4 3-·l-1).
Previous references have also related this
species with surrounding arable lands and
nutrient enrichment (Johansson, 1982), and
also with high N:P ratios (Stancheva et al.,
2013; Aboal et al., 2014). However, this species
also appears under nitrate-reduced conditions.
Therefore, it could be a good indicator of
absence of urban impacts in our study area, but
not a good indicator of agricultural impact or
nitrate enrichment given its high level of
tolerance to this nutrient.
References on T. distorta to low nutrient
contents (mainly total phosphorus or
phosphate) are frequent (e.g. Schneider and
Melzer, 2003; Kastovsky et al., 2010; Mateo et
al., 2010). Even the genus Tolypothrix is
described as being an inhabitant of clear
unpolluted waters (Wehr and Sheath, 2003).
However in our study area, T. distorta grew at
sites with low phosphate concentrations, but in
agricultural areas, and could be an indicator of
agricultural impact. This fact reinforces the
statement that the ecological ranges of species
should be studied regionally prior to
establishing the indicator potential of a
particular species, as suggested by Rott and
Schneider (2014).
5. CONCLUSIONS
(1) In the study area, the most influential
variable on species composition was
conductivity. Among the considered nutrients,
101
phosphate had a stronger influence than
nitrate.
(2) Nostoc verrucosum, Phormidium autumnale,
Plectonema tomasinianum, Rivularia haematites
and Tolypothrix distorta have been found to be
sensitive taxa to nutrient contents, and could
be suitable indicators of anthropogenic
pressures. Therefore, they could be useful tools
for developing biological water quality indices.
(3) Regional studies on the ecological ranges
and optima of species are desirable prior to
selecting those appropriate for bioindications.
(4) The suitability of some species as
bioindicators may vary from one different
geographic region to another.
(5) Further research into the indicator
potential of benthic cyanobacteria is desirable
to use them to improve the accuracy of
biological indices to assess the ecological status
of rivers according to the Water Framework
Directive.
Acknowledgements:
This study has been supported by several
projects granted by the Regional Government
of Castilla-La Mancha (Spain): PO1109-0190-
8090, PPII10-0271-1349, PREG01-0016 and
PREG06-027.
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Demars, B.O.L., Potts, J.M., TréMolièRes, M., ThiéBaut,
G., Gougelin, N., Nordmann, V., 2012. River
macrophyte indices: not the Holy Grail!: River
macrophyte indices: an enticing mirage. Freshw.
Biol. 57, 1745–1759. doi:10.1111/j.1365-
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Ennis, G.L., 1978. Algal indicators of water quality in
the Flathead River Basin. J PhycolUnited States 14.
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Albuquerque, A., 2005. Assessing biotic integrity in
Iberian rivers: Development of a multimetric plant
index. Ecol. Indic. 5, 137–149.
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Jafari, N. G., & Gunale, V. R., 2006. Hydrobiological study
of algae of an urban freshwater river. J. Appl. Sci.
Environ. Manag. 10 (2).153–158.Johansson, C.,
1982. Attached algal vegetation in running waters of
Jämtland, Sweden.
Kastovsky, J., Hauer, T., Komarek, J., Skacelova, O., 2010.
The list of cyanobacterial species of the Czech
Republic to the end of 2009. Fottea 245–249.
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Komárek, J., Anagnostidis, K., 2005. Süßwasserflora von
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Larkum, A.W.D., Kennedy, I.R., Muller, W.J., 1988.
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Lee, R. E. (2008). Phycology. Cambridge
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Loza, V., Berrendero, E., Perona, E., Mateo, P., 2013.
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molecular, and ecological approaches 1. J. Phycol. 49,
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Loza, V., Perona, E., Mateo, P., 2014. Specific responses
to nitrogen and phosphorus enrichment in
cyanobacteria: Factors influencing changes in
species dominance along eutrophic gradients.
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Lukavský, J., Moravcová, A., Nedbalová, L., Rauch, O.,
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streams in the Bohemian Forest under the influence
of recreational activity. Biologia (Bratisl.) 61.
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dulces del noroeste de España. Publicaciones Inst.
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phenotypic diversity of cyanobacteria assigned to
the genus Phormidium (Oscillatoriales) from
different habitats and geographical sites. Arch.
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Mateo, P., Berrendero, E., Perona, E., Loza, V., Whitton,
B.A., 2010. Phosphatase activities of cyanobacteria
as indicators of nutrient status in a Pyrenees river.
Hydrobiologia 652, 255–268. doi:10.1007/s10750-
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Mollenhauer, D., Bengtsson, R., Lindstrøm, E.-A., 1999.
Macroscopic cyanobacteria of the genus Nostoc: a
neglected and endangered constituent of European
inland aquatic biodiversity. Eur. J. Phycol. 34, 349–
360. doi:10.1080/09670269910001736412
Monteagudo, L., Moreno, J.L., Picazo, F., 2012. River
eutrophication: Irrigated vs. non-irrigated
agriculture through different spatial scales. Water
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doi:10.1016/j.watres.2012.02.035
Moreno, J.L., Aboal, M., Monteagudo, L., 2012. On the
presence of Nostochopsis lobata Wood ex Bornet et
Flahault in Spain: morphological, ecological and
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on heterocyst production in blue-green algae.
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DISCUSIÓN GENERAL
Arroyo de Nava del Rey, Huertezuelas (Ciudad Real)
Fotografía: José Luis Moreno Alcaraz

103
104
6. DISCUSIÓN GENERAL

Los resultados de esta Tesis Doctoral, muestran que Castilla-La Mancha es una
región rica en macroalgas. En general, los filos Cyanophyta (=Cyanobacteria) (algas
verdeazules), Rhodophyta (algas rojas) y Chlorophyta (algas verdes) están
ampliamente distribuidos en el área de estudio y cuentan con numerosos
representantes. Muchos de ellos están mundialmente extendidos, mientras que
algunas especies han sido documentadas de forma ocasional en Europa. Estas
especies raras han sido: Nostochopsis lobata Wood ex Bornet et Flahaut,
Batrachospermum atrum (Hudson) Harvey, Chroothece richteriana Hansgirg,
Oocardium stratum Nägeli y Tetrasporidium javanicum Möbius. Sin embargo, con la
revisión en el futuro de todo el material recolectado a nivel específico, no se descarta
la aparición de más especies de interés regional, nacional o europeo. En concreto N.
lobata únicamente ha sido detectada en cuatro localidades europeas, localizadas en
cuatro países (Italia, Francia, Portugal y España), y en sólo una única ocasión por
localidad. Sin embargo, esta especie es muy abundante en la zona tropical, donde
desarrolla colonias de mayor tamaño que las descritas en Europa. Tal y como se
refleja en el Artículo 5.3., las citas de esta especie tropical en la zona templada han
aumentado en los últimos años. A pesar de no ser una especie apta para la
bioindicación de presiones antropogénicas debido a su escasez en
nuestro continente, sí podría ser útil como indicadora de calentamiento global. De
la misma manera, la inestabilidad de poblaciones de Hydrurus foetidus (Vilars)
Trevisan, en algunas zonas montañosas, también puede ser un efecto de cambios
en el clima, ya que es una especie estenoterma, mundialmente extendida, que
se desarrolla en aguas frías dentro de un rango muy reducido de temperatura
(entre 2 y 12°C).

En cuanto al resto de taxones, la mayoría presentan una distribución

geográfica amplia, tal y como se espera en especies indicadoras. Sin embargo, sólo
unos pocos de ellos han sido incluidos en algún índice de macrófitos. En la
comparación de métricas realizada por Demars et al. (2012), se observó que menos
de la mitad de los índices incluían macroalgas y, de hacerlo, se consideraban
preferentemente a nivel de género, a pesar de que la diversidad ecológica
intragenérica puede ser muy

105
elevada. Todo ello indica que mejorar la precisión de las métricas basadas en
macrófitos sería posible si se incluyeran macroalgas indicadoras a nivel de especie.

Otro de los requisitos que debe cumplir una especie indicadora es, obviamente, que
sea capaz de aportar información sobre los impactos que afectan al medio en el que
vive, por lo que el estudio de estas presiones es un punto clave en la búsqueda de
especies indicadoras. Una de las contribuciones más relevantes de este trabajo tiene
que ver con el análisis del impacto de los usos de suelo en la calidad del agua y, más
concretamente, en cómo analizar este factor de la manera más adecuada. En Castilla-
La Mancha las principales presiones sobre los ríos y arroyos son la agricultura
(principal fuente de contaminación difusa) y los vertidos de aguas residuales
urbanas (contaminación puntual). Según los resultados obtenidos en el Artículo 5.4.,
la agricultura aparece como la causa principal de eutrofización en los sistemas
fluviales, en concordancia con otros estudios realizados previamente en la zona
(Moreno et al., 2006; Lassaletta et al., 2009). Comparando las diferentes técnicas
agrícolas, los resultados indicaron que el regadío es la que más impacto causa en el
sistema fluvial castellano-manchego, a pesar de ocupar una superficie mucho menor
que el secano. Además, se detectó un umbral de impacto que sugiere que al superar
un 50% de suelo dedicado al cultivo en regadío en zonas muy cercanas a los cauces
(1km), se incrementa considerablemente el riesgo de eutrofización. Este hecho
apoya la idea de que la intensidad del uso agrícola puede ser un mejor indicador del
impacto que el porcentaje de suelo que ocupa (Harding et al., 1999).

En el caso del regadío, la superficie que ocupa es mucho menor que la del secano
por lo que, empleando escalas espaciales amplias (p. ej. área total de drenaje),
el secano enmascara los resultados y no es posible detectar el efecto del regadío.
Para evitar este efecto, se testaron diversas escalas, y los resultados mostraron que
la más adecuada fue la zona de influencia de 1km de radio aguas arriba, ya que
con ella se obtiene el gradiente de datos más completo para todos los usos de suelo
y se evita el problema de la autocorrelación espacial (Figura 4). Estos resultados
subrayan que la escala de estudio afecta a los resultados y que, tal y como sugiere
Chang (2008) es necesario considerar los enfoques ‘multiescala’ para poder aclarar
la dinámica de la calidad del agua en el tiempo y el espacio. La contribución más
novedosa del Artículo

106
5.4. es la propuesta de una metodología a seguir para la elección de la escala más
adecuada en este tipo de estudios.

Figura 4. Elección de la escala espacial adecuada al caso de estudio.

Una vez estudiada la diversidad de macroalgas e identificadas las presiones se

procedió a analizar cómo afectan las variables ambientales a la comunidad de
macroalgas (Artículo 5.5.). En este caso, los usos de suelo se contabilizaron a través
de áreas de influencia de 1 km de radio aguas arriba, tal y como se estableció en el
trabajo anterior. El grupo de algas elegido como objeto de estudio fue el de las
cianobacterias, ya que estaban ampliamente representadas y fueron detectadas en
una gran variedad de ambientes (zonas con impacto, aguas puras, etc.). Según los
resultados obtenidos, la conductividad fue la variable más determinante en la
composición de especies de la comunidad de cianobacterias (Figura 5). Una vez
descartado su efecto, se pudo observar el efecto de los nutrientes, fosfato y nitrato,
por orden de importancia. Tal y como apunta Biggs (1990), esto puede deberse a
que la conductividad es un factor que integra otros factores de gran envergadura
ecológica que actúan a gran escala, tales como litología, clima, etc. Aunque no se
encontraron grandes diferencias entre cianobacterias con heterocitos y sin ellos, las
primeras fueron más frecuentes en los sitios con un nivel de nitrato bajo, tal y como

107
se esperaba y como se ha descrito en otras zonas geográficas (p. ej. Stancheva et al.,
2013).

Figura 5. Factores determinantes y especies típicas en las comunidades de cianobacterias.

Observando los rangos ecológicos de las especies y dados los resultados de análisis
de la composición de las comunidades según las condiciones ambientales, se
pudieron identificar cinco especies adecuadas para la bioindicación de las presiones
humanas. Nostoc verrucosum Vaucher ex Bornet et Flahault parece indicar de
ausencia de presiones, de acuerdo con descripciones previas de la ecología de la
especie (Ennis, 1978; Mollenhauer et al., 1999). Phormidium autumnale Gomont se
encontró en sitios impactados tanto por vertidos urbanos como por agricultura, en
concordancia con los estudios que relacionan esta especie con aguas contaminadas
(Barinova et al., 2010; Loza et al., 2013). Tanto Plectonema tomasinianum Bornet ex
Gomont como Tolypothrix distorta Kützing ex Bornet et Flahault aparecen

108
relacionados en la literatura con aguas oligotróficas (p. ej Schneider and Melzer,
2003 y Kastovsky et al., 2010, respectivamente). Sin embargo, en el área de estudio
P. tomasinianum parece indicar impacto urbano y T. distorta impacto agrícola, lo que
apoya la teoría sugerida por Rott y Schneider (2014) de que antes de establecer el
potencial indicador de una especie es necesario estudiar los rangos ecológicos de la
misma a nivel regional. Por último, Rivularia haematites C. Agardh ex Bornet et
Flahault mostró predilección por los sitios con alto nivel de nitrato, de acuerdo con
trabajos previos que relacionan esta especie con ratios elevados de N:P (Stancheva
et al., 2013; Aboal et al., 2014). Sin embargo, no puede considerarse un buen
indicador de impacto agrícola ya que también se desarrolla en lugares pobres en
nitrato. Por otro lado, su tolerancia al fosfato fue muy reducida, por lo que sí puede
considerarse un buen indicador de ausencia de impacto urbano.

Tanto la metodología establecida para el estudio de las presiones como el enfoque

aplicado en la evaluación del potencial indicador de las cianobacterias, pueden
servir de base para identificar más especies de algas indicadoras en ésta y otras
áreas geográficas. Ampliar el abanico de bioindicadores disponibles podrá
contribuir, en un futuro, al perfeccionamiento de los índices biológicos de calidad del
agua empleados en aplicación de la Directiva Marco De Agua (2000/60/CE).

109
110
CONCLUSIONES
Río Júcar en Las Mariquillas (Albacete)
Fotografía: José Luis Moreno Alcaraz

111
112
7. CONCLUSIONES

De los resultados obtenidos en los trabajos comprendidos en esta Tesis Doctoral, se

desprenden las siguientes conclusiones, agrupadas según las líneas de trabajo
planteadas en la misma:

7.1. Diversidad y distribución de las macroalgas bentónicas de Castilla-La

Mancha:

 Castilla-La Mancha es una región con una gran diversidad de macroalgas

fluviales. Los filos Cyanophyta (=Cyanobacteria) y Chlorophyta son los más
frecuentes y más ampliamente distribuidos.
 Hasta el momento, la región cuenta con la presencia de cinco especies
consideradas ‘raras’ a nivel europeo: Nostochopsis lobata Wood ex Bornet et
Flahaut, Batrachospermum atrum (Hudson) Harvey, Chroothece richteriana
Hansgirg, Oocardium stratum Nägeli y Tetrasporidium javanicum Möbius.
 El hallazgo de Nostochopsis lobata en el Arroyo de Nava del Rey (Ciudad
Real), supone la primera cita nacional publicada de esta especie y la cuarta
europea.
 Los ejemplares de Hydrurus foetidus (Villars) Trevisan encontrados en el
Parque Natural del Hayedo de Tejera Negra (Guadalajara), suponen la
primera cita de esta especie en la región.

7.2. Presiones antropogénicas en el área de estudio:

 El regadío es la principal causa de eutrofización en el sistema fluvial

castellano-manchego. Lo que indica que tanto la intensidad de la práctica
agrícola como su proximidad a los cauces son factores importantes a tener en
cuenta.
 Sobrepasar el umbral de presión del 50% del suelo dedicado al regadío en un
radio de influencia de 1km en ambas márgenes de los ríos, puede ocasionar
un aumento rápido de la concentración de nitrato y, por tanto, eutrofización
de las aguas.

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  La elección de la escala espacial puede afectar a los resultados en estudios
que relacionan los usos de suelo con la calidad del agua y, por tanto, puede
influir también en la interpretación ecológica de los mismos.
 En el área de estudio, la escala espacial más adecuada para analizar el
impacto de la agricultura fue el radio de influencia de 1km aguas arriba de las
estaciones de muestreo.
 La escala más adecuada puede variar según el objeto de estudio y, en
ocasiones, puede haber incluso más de una escala espacial que proporcione
resultados reales y fiables.
 Como contribución novedosa, la siguiente metodología puede servir de base
en la búsqueda de la escala más apropiada para el estudio de los usos de
suelo como causantes de la eutrofización en ríos y arroyos:
a) Es importante analizar qué escala(s) proporciona(n) el rango o
gradiente más amplio de datos para cada clase de uso de suelo que se
estudia. Cuanto más amplio sea este rango, más fiables serán los
resultados obtenidos.
b) Es necesario comprobar que no exista autocorrelación espacial de los
datos, algo frecuente en distancias geográficas reducidas.
c) La escala más adecuada será aquella que nos permita trabajar con la
muestra más completa, sin generar autocorrelación.

7.3. Relación entre la comunidad de cianobacterias y la calidad del agua:

 En el área de estudio, la conductividad es la variable más determinante en la

composición de especies de la comunidad de cianobacterias. Comparando los
nutrientes estudiados, el fosfato influye más en la comunidad que el nitrato.
 Para evaluar el potencial indicador de una determinada especie es
conveniente analizar previamente sus rangos de tolerancia y sus óptimos
ecológicos a nivel regional.
 El potencial indicador de una determinada especie puede variar de una zona
geográfica a otra.
 Nostoc verrucosum Vaucher ex Bornet et Flahault, Phormidium autumnale
Gomont, Plectonema tomasinianum Bornet ex Gomont, Rivularia haematites C.

114
Agardh ex Bornet et Flahault y Tolypothrix distorta Kützing ex Bornet et
Flahault han mostrado ser especies sensibles a los niveles de nutrientes y
tener potencial como bioindicadores de las presiones antropogénicas. Por
tanto, podrían tenerse en cuenta en el desarrollo de índices biológicos.

115
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BIBLIOGRAFÍA
Tapetes de Phormidium sp.
Fotografía: José Luis Moreno Alcaraz

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118
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 AGRADECIMIENTOS

Durante todos estos años, ha sido mucha la gente que me ha tendido la mano y ha
contribuido, de una forma u otra, a que hoy esté escribiendo las últimas líneas de
esta Tesis. Os quiero dar las gracias a todos por haber formado parte de esta etapa
de mi vida, espero no dejarme a nadie en el tintero.

La primera persona que me viene a la mente eres tú, José Luis. Muchas gracias por
todo esto que hoy queda materializado en papel, por haber hecho siempre todo lo
posible y más para ayudarme, por contagiarme de ilusión y esperanza siempre que
lo he necesitado, por tu comprensión… pero sobre todo, por tu generosidad. Has
compartido conmigo tu tiempo, conversaciones geniales, recursos de todo tipo,
sabiduría y ¡hasta alguna lección de conducir!. Me quedo con muchas cosas de ti que
me acompañaran siempre. Que conste que esto no es una despedida, ¡aún te debo
unos cuántos mapas!.

Gracias a mis padres. Aunque se me encoja un poco el corazón pensando que no

leeréis estas líneas, me consuela pensar que de alguna forma os llega el mensaje.
Gracias por absolutamente todo. Me enseñasteis lo más importante en esta vida: que
un hogar no tiene nada que ver con cemento ni ladrillos, que tiene que ver con el
amor, y el amor es eterno. Y precisamente por eso, gracias también a mis hermanos,
Alberto y Ricardo, porque siempre y en cualquier parte del mundo, estando con
vosotros, estaré en casa.

Jose, gracias por recordarme que la vida es muy sencilla siempre que se me olvida.
Por compartir conmigo miles de pequeños momentos que me llenan el corazón y por
hacerme reír. Me alegro de no haber hecho caso a aquello de ‘no te eches un novio
murciano que son muy mentirosos y muy negociantes’ jejeje. Y, cómo no, gracias al
pequeño Mojo que me regala el sol, el aire y la lluvia sacándome a pasear todos
los días, y que es capaz de hacer desaparecer un mal día haciéndome sentir la
persona más importante del mundo. Todo es mejor con vosotros dos.

Gran parte del impulso necesario para llegar hasta aquí me lo ha dado mi familia
pricense al completo, que es un tesoro. Sobretodo tú Tita, que eres experta en

125
querer y cuidar a los que te rodean. Me alegro mucho de poder decirte que esto ya
está.

Gracias a mis amigos, que son geniales… En especial a Myriam, Lourdes y Milla,
porque sois personas excepcionales (literalmente, dudo mucho que abunde la gente
como vosotros). Sois la mejor compañía que una podría tener en esta vida y es un
privilegio poder contar con vosotros siempre. Cuanto más os conozco, más os
admiro y os quiero. Y a Estela, qué buenas esas tardes de cañas y de buen cine, el
mejor aderezo para estos años de trabajo.

Al grupo de los miércoles, gracias por haber compartido tanto con mi madre y con
nosotros. Habéis llenado su memoria y la nuestra de muy buenos recuerdos, de
risas, viajes y un sinfín de anécdotas. Gracias por haber sido nuestros ángeles de la
guarda, sin vosotras todo hubiera sido mucho más difícil.

Gracias a todas las personas que formaron parte del Área de Limnología Aplicada e
Hidrobiología antes de mi llegada y cuyo trabajo y esfuerzo forma parte de esta
memoria.

Al grupo de Ecología Acuática de la Universidad de Murcia, mil gracias por acogerme

como una más en diversas ocasiones académico-sociales, espero poder veros en
muchas otras… me suena que algo se cuece en Villalgordo.

A Pedro Tomás, gracias por invitarme a compartir con vosotros aquellos días de
muestreo en Huesca y ¡enhorabuena por tu Tesis!.

Por último, gracias a David Sánchez y Daniel Moya por su amabilidad revisando este
trabajo.

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