Edición Bilingüe

Bilingual Edition
ISSN: 2448-6698
Revista Mexicana de Ciencias Pecuarias Rev. Mex. Cienc. Pecu. Vol. 14 Núm. 3, pp. 488-744, JULIO-SEPTIEMBRE-2023

Rev. Mex. Cienc. Pecu. Vol. 14 Núm. 3, pp. 488-744, JULIO-SEPTIEMBRE-2023

 REVISTA MEXICANA DE CIENCIAS PECUARIAS Volumen 14 Numero 3, Julio-
Septiembre 2023. Es una publicación trimestral de acceso abierto, revisada por pares y
arbitrada, editada por el Instituto Nacional de Investigaciones Forestales, Agrícolas y
Pecuarias (INIFAP). Avenida Progreso No. 5, Barrio de Santa Catarina, Delegación Coyoacán,
C.P. 04010, Cuidad de México, www.inifap.gob.mx.
Distribuida por el Centro de Investigación Regional Sureste, Calle 6 No. 398 X 13, Avenida
Correa Racho, Col. Díaz Ordaz, Mérida Yucatán, C.P. 97130.
Editor responsable: Arturo García Fraustro Reservas de Derechos al Uso Exclusivo número
04-2022-033116571100-102, ISSN: 2448-6698, otorgados por el Instituto Nacional del
Derecho de Autor (INDAUTOR).
Responsable de la última actualización de este número: Arturo García Fraustro, Campo
Experimental Mocochá, Km. 25 Antigua Carretera Mérida–Motul, Mocochá, Yuc. C.P. 97454.
http://cienciaspecuarias. inifap.gob.mx, la presente publicación tuvo su última actualización
en julio de 2023.
Campesina observando formas parasitarias
al microscopio en los laboratorios
interactivos; Rancho ovino pelibuey “La
Finquita”, municipio Cauto Cristo Granma, DIRECTORIO
Cuba.
Autor: Manuel Alejandro La O Arias FUNDADOR
John A. Pino
EDITOR EN JEFE EDITORES ADJUNTOS
Arturo García Fraustro Oscar L. Rodríguez Rivera
Alfonso Arias Medina
EDITORES POR DISCIPLINA

Dra. Yolanda Beatriz Moguel Ordóñez, INIFAP, México
Dr. Ramón Molina Barrios, Instituto Tecnológico de Sonora,
Dr. Alfonso Juventino Chay Canul, Universidad Autónoma de
Tabasco, México
Dra. Maria Cristina Schneider, Universidad de Georgetown,
Estados Unidos
Dr. Feliciano Milian Suazo, Universidad Autónoma de
Querétaro, México
Dr. Javier F. Enríquez Quiroz, INIFAP, México
Dra. Martha Hortencia Martín Rivera, Universidad de Sonora
URN, México
Dr. Fernando Arturo Ibarra Flores, Universidad de Sonora
URN, México
Dr. James A. Pfister, USDA, Estados Unidos
Dr. Eduardo Daniel Bolaños Aguilar, INIFAP, México
Dr. Sergio Iván Román-Ponce, INIFAP, México
Dr. Jesús Fernández Martín, INIA, España
Dr. Maurcio A. Elzo, Universidad de Florida
Dr. Sergio D. Rodríguez Camarillo, INIFAP, México
Dra. Nydia Edith Reyes Rodríguez, Universidad Autónoma del
Estado de Hidalgo, México
Dra. Maria Salud Rubio Lozano, Facultad de Medicina
Veterinaria y Zootecnia, UNAM, México
Dra. Elizabeth Loza-Rubio, INIFAP, México
Dr. Juan Carlos Saiz Calahorra, Instituto Nacional de
Investigaciones Agrícolas, España
Dr. José Armando Partida de la Peña, INIFAP, México
Dr. José Luis Romano Muñoz, INIFAP, México
Dr. Jorge Alberto López García, INIFAP, México
Dr. Alejandro Plascencia Jorquera, Universidad Autónoma de
Baja California, México
Dr. Juan Ku Vera, Universidad Autónoma de Yucatán, México
Dr. Ricardo Basurto Gutiérrez, INIFAP, México
Dr. Luis Corona Gochi, Facultad de Medicina Veterinaria y
Zootecnia, UNAM, México
Dr. Juan Manuel Pinos Rodríguez, Facultad de Medicina
Veterinaria y Zootecnia, Universidad Veracruzana, México
Dr. Carlos López Coello, Facultad de Medicina Veterinaria y
Zootecnia, UNAM, México
Dr. Arturo Francisco Castellanos Ruelas, Facultad de
Química. UADY
Dra. Guillermina Ávila Ramírez, UNAM, México
Dr. Emmanuel Camuus, CIRAD, Francia.
Dr. Héctor Jiménez Severiano, INIFAP., México
Dr. Juan Hebert Hernández Medrano, UNAM, México
Dr. Adrian Guzmán Sánchez, Universidad Autónoma
Metropolitana-Xochimilco, México
Dr. Eugenio Villagómez Amezcua Manjarrez, INIFAP, CENID
Salud Animal e Inocuidad, México
Dr. José Juan Hernández Ledezma, Consultor privado
Dr. Fernando Cervantes Escoto, Universidad Autónoma
Chapingo, México
Dr. Adolfo Guadalupe Álvarez Macías, Universidad Autónoma
Metropolitana Xochimilco, México
Dr. Alfredo Cesín Vargas, UNAM, México
Dra. Marisela Leal Hernández, INIFAP, México
Dr. Efrén Ramírez Bribiesca, Colegio de Postgraduados,
México
Dra. Itzel Amaro Estrada, INIFAP, México

TIPOGRAFÍA Y FORMATO: Oscar L. Rodríguez Rivera

Indizada en el “Journal Citation Report” Science Edition del ISI . Inscrita en el Sistema de Clasificación de Revistas Científicas y
Tecnológicas de CONACyT; en EBSCO Host y la Red de Revistas Científicas de América Latina y el Caribe, España y Portugal
(RedALyC) (www.redalyc.org); en la Red Iberoamericana de Revistas Científicas de Veterinaria de Libre Acceso
(www.veterinaria.org/revistas/ revivec); en los Índices SCOPUS y EMBASE de Elsevier (www.elsevier. com).

I
 REVISTA MEXICANA DE CIENCIAS PECUARIAS
La Revista Mexicana de Ciencias Pecuarias es un órgano
de difusión científica y técnica de acceso abierto, revisada
por pares y arbitrada. Su objetivo es dar a conocer los
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Todas las contribuciones serán cuidadosamente evaluadas
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VISIT OUR SITE IN THE INTERNET
Full articles from year 1963 to date and Instructions for authors can be accessed via the site http://cienciaspecuarias.inifap.gob.mx
II
I
trimestral en formato bilingüe Español e Inglés. El costo
total por publicar es de $ 7,280.00 más IVA por manuscrito
ya editado.
Se publica en formato digital en acceso abierto, por lo que
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.
 REVISTA MEXICANA DE CIENCIAS PECUARIAS

REV. MEX. CIENC. PECU. VOL. 14 No. 3 JULIO-SEPTIEMBRE-2023

CONTENIDO
Contents

ARTÍCULOS
Articles
Pág.

Predicción de la composición química de las heces y digesta ileal de cerdos mediante

espectroscopia de reflectancia en el infrarrojo cercano (NIRS)
Prediction of the chemical composition of pig feces and ileal digesta by near-infrared reflectance
spectroscopy (NIRS)
Ricardo Ramos Cruz, Ricardo Basurto Gutiérrez, Ericka Ramírez Rodríguez, Tércia Cesária Reis de
Souza, Gerardo Mariscal Landín .............................................................................................488

Effect of hesperidin supplementation on blood profile, antioxidant capacity, intestinal

histomorphology and fecal microbial counts in Japanese quails
Efectos de la suplementación con hesperidina en la codorniz japonesa Cordonix cordonix japonica
sobre la química sanguínea, la capacidad antioxidante, la histopatología intestinal y la microflora
fecal
Abdullah Özbilgin, Mahmut Niyazi Moğulkoç, Füsun Erhan Bayçumendur, Nazlı Ercan.................505

Supplementation of goats with mesquite pod meal in deferred ( Urochloa

mosambicenses) grass pasture in the semiarid region
Suplementación de caprinos con harina de vaina de mezquite en una pradera diferida de pasto
(Urochloa mosambicensis) en la región semiárida
Diego da Hora Souto, Mara Lúcia Albuquerque Pereira, Taiala Cristina de Jesus Pereira, Herymá
Giovane de Oliveira Silva, Paulo José Presídio Almeida, Leandro Borges Sousa, Fernando Oliveira
Barreto, Larisse Borges Sousa, Karine Pinheiro Oliveira, Gleidson Giordano Pinto de Carvalho ....523

Parámetros, correlaciones y tendencias genéticas de caracteres reproductivos en

ganado Holstein de México
Genetic parameters, correlations and trends of reproductive traits in Holstein cattle from Mexico
Carolina Durán-Alvarez, Adriana García-Ruiz, Rogelio Alejandro Alonso Morales, Luis E. Eguiarte,
Felipe de Jesús Ruiz-López ....................................................................................................539

III
Efectos genéticos aditivos y no aditivos para características reproductivas en dialelo
Holstein-Suizo Pardo en clima subtropical húmedo
Additive and non-additive genetic effects for reproductive traits in a Holstein- Brown Swiss diallel in
humid subtropical climate
Luis Adrián García Bravo, Vicente Eliezer Vega-Murillo, José Alfredo Villagómez- Cortes, Juan
Prisciliano Zárate-Martínez, Otto Raúl Leyva-Ovalle, René Carlos Calderón Robles, Ángel Ríos-
Utrera, Moisés Montaño Bermúdez, Guillermo Martínez-Velázquez ............................................556

Dinámica del pastoreo en la asociación cultivos y ovinos de agroecosistemas de clima

templado en México
Dynamics of grazing in the association of crops and sheep of temperate climate agroecosystems in
Mexico
Samuel Vargas López, Angel Bustamante González, José Luis Jaramillo Villanueva, Ignacio Vázquez
Martínez, Humberto Vaquera Huerta, Carla Cristina Díaz Sánchez, Miguel Ángel Casiano
Ventura……………………………………………………………………………………………………………………………..572

Laboratorios interactivos para el aprendizaje experiencial y el manejo integrado del

parasitismo gastrointestinal de ovinos y caprinos
Interactive laboratories for experiential learning and integrated management of gastrointestinal
parasitism of sheep and goats
Manuel Alejandro La O-Arias, Francisco Guevara-Hernández, José Roberto Aguilar-Jiménez, René
Pinto-Ruíz, Luis Reyes-Muro, José Nahed-Toral .......................................................................592

Sincronización de estros en ovejas mediante protocolo de 6 días con CIDR de primera,

segunda y tercera reutilización
Estrus synchronization in ewes with a six-day protocol using new, second-use, third-use and fourth-
use CIDR devices
Susana López-García, María Teresa Sánchez-Torres Esqueda, José Luis Cordero-Mora, José Luis
Figueroa-Velasco, José Alfredo Martínez-Aispuro, Teódulo Salinas Ríos .....................................610

Evaluación de indicadores de bienestar animal de bovinos en un rastro Tipo Inspección

Federal
Evaluation of animal welfare indicators of cattle in a Federal Inspection Type slaughterhouse
Jaime Noé Sánchez-Pérez, Horacio Dávila-Ramos, Juan Carlos Robles-Estrada, Jesús José Portillo-
Loera, Armida Sánchez-Escalante, Daniel Diaz ........................................................................622

Evaluación in vitro del potencial acaricida de Beauveria bassiana DS3.17 sobre la

garrapata común (Rhipicephalus microplus) en Oaxaca, México
In vitro evaluation of the acaricidal potential of Beauveria bassiana DS3.17 on the common tick
(Rhipicephalus microplus) in Oaxaca, Mexico
Jared Martínez-García, José Abad-Zavaleta, María de Jesús García-Gómez, Oscar Núñez-Gaona…644

Análisis de la disponibilidad a pagar por carne de cerdo libre de antibióticos, un enfoque

de Experimentos de Elección
Analysis of the willingness to pay for antibiotic-free pork, a Choice Experiments approach
Miriam Susana Hernández Valdivia, Enrique Melo Guerrero, Ramón Valdivia Alcalá, Luis Manuel
Valenzuela Núñez, Juan Hernández Ortiz, Miguel Ángel Martínez Damián ..................................658

IV
REVISIONES DE LITERATURA
Reviews

Proteína Bm86 y su potencial uso como vacuna contra garrapatas en el ganado bovino.
Revisión
Bm86 protein and its potential use as an anti-tick vaccine in cattle. Review
Dulce María Galván Arellano, Patricia Vieyra Reyes, Roberto Montes de Oca Jiménez, Jaime Ortega
Lopez, Saúl Gabriel Martínez Arzate, Bruno Rivas Santiago, Juan Carlos Vázquez Chagoyán .......672

Principios y prácticas agroecológicas para la transición hacia una ganadería bovina

sostenible. Revisión
Agroecological principles and practices for the transition to sustainable cattle farming. Review
Guadalupe Del Angel-Lozano, Miguel Ángel Escalona-Aguilar, Julio Baca del Moral, Venancio
Cuevas-Reyes .......................................................................................................................696

NOTAS DE INVESTIGACIÓN
Technical notes

Suplementación de quitosano en la dieta de pollitas Rhode Island Red y su efecto en el

comportamiento productivo y variables hematológicas
Chitosan supplementation in the diet of Rhode Island Red pullets and its effect on productive
behavior and hematological variables
Edgar Fernando Peña-Torres, José Abel Ortega, Cynthia Esmeralda Lizárraga-Velázquez,
Crisantema Hernández, Jesús Armando León Cañedo, Asahel Benitez-Hernández ......................725

Función Cobb-Douglas de la producción de miel en Aguascalientes, México

Analyzing honey production in Aguascalientes, Mexico, using the Cobb-Douglas Function
Marco Andrés López Santiago, José Inés Zavala Beltrán, Ramón Valdivia Alcalá, Blanca Margarita
Montiel Batalla ......................................................................................................................735

V

NOTAS AL AUTOR
La Revista Mexicana de Ciencias Pecuarias se edita
completa en dos idiomas (español e inglés) y publica tres
categorías de trabajos: Artículos científicos, Notas de
investigación y Revisiones bibliográficas.
Los autores interesados en publicar en esta revista
deberán ajustarse a los lineamientos que más adelante se
indican, los cuales en términos generales, están de
acuerdo con los elaborados por el Comité Internacional de
Editores de Revistas Médicas (CIERM) Bol Oficina Sanit
Panam 1989;107:422-437.
1. Sólo se aceptarán trabajos inéditos. No se admitirán
si están basados en pruebas de rutina, ni datos
experimentales sin estudio estadístico cuando éste
sea indispensable. Tampoco se aceptarán trabajos
que previamente hayan sido publicados condensados
o in extenso en Memorias o Simposio de Reuniones o
Congresos (a excepción de Resúmenes).
2. Todos los trabajos estarán sujetos a revisión de un
Comité Científico Editorial, conformado por Pares de
la Disciplina en cuestión, quienes desconocerán el
nombre e Institución de los autores proponentes. El
Editor notificará al autor la fecha de recepción de su
trabajo.
3. El manuscrito deberá someterse a través del portal de
la Revista en la dirección electrónica:
http://cienciaspecuarias.inifap.gob.mx, consultando
el “Instructivo para envío de artículos en la página de
la Revista Mexicana de Ciencias Pecuarias”. Para su
elaboración se utilizará el procesador de Microsoft
Word, con letra Times New Roman a 12 puntos, a
doble espacio. Asimismo se deberán llenar los
formatos de postulación, carta de originalidad y no
duplicidad y disponibles en el propio sitio oficial de la
revista.
4. Por ser una revista con arbitraje, y para facilitar el
trabajo de los revisores, todos los renglones de cada
página deben estar numerados; asimismo cada
página debe estar numerada, inclusive cuadros,
ilustraciones y gráficas.
5. Los artículos tendrán una extensión máxima de 20
cuartillas a doble espacio, sin incluir páginas de Título,
y cuadros o figuras (los cuales no deberán exceder de
ocho y ser incluidos en el texto). Las Notas de
investigación tendrán una extensión máxima de 15
cuartillas y 6 cuadros o figuras. Las Revisiones
bibliográficas una extensión máxima de 30 cuartillas y
5 cuadros.
VI
Actualización: marzo, 2020
6. Los manuscritos de las tres categorías de trabajos que
se publican en la Rev. Mex. Cienc. Pecu. deberán
contener los componentes que a continuación se
indican, empezando cada uno de ellos en página
aparte.
Página del título
Resumen en español
Resumen en inglés
Texto
Agradecimientos y conflicto de interés
Literatura citada
7. Página del Título. Solamente debe contener el título
del trabajo, que debe ser conciso pero informativo; así
como el título traducido al idioma inglés. En el
manuscrito no es necesaria información como
nombres de autores, departamentos, instituciones,
direcciones de correspondencia, etc., ya que estos
datos tendrán que ser registrados durante el proceso
de captura de la solicitud en la plataforma del OJS
(http://ciencias pecuarias.inifap.gob.mx).
8. Resumen en español. En la segunda página se debe
incluir un resumen que no pase de 250 palabras. En
él se indicarán los propósitos del estudio o
investigación; los procedimientos básicos y la
metodología empleada; los resultados más
importantes encontrados, y de ser posible, su
significación estadística y las conclusiones principales.
A continuación del resumen, en punto y aparte,
agregue debidamente rotuladas, de 3 a 8 palabras o
frases cortas clave que ayuden a los indizadores a
clasificar el trabajo, las cuales se publicarán junto con
el resumen.
9. Resumen en inglés. Anotar el título del trabajo en
inglés y a continuación redactar el “abstract” con las
mismas instrucciones que se señalaron para el
resumen en español. Al final en punto y aparte, se
deberán escribir las correspondientes palabras clave
(“key words”).
10. Texto. Las tres categorías de trabajos que se publican
en la Rev. Mex. Cienc. Pecu. consisten en lo
siguiente:
a) Artículos científicos. Deben ser informes de trabajos
originales derivados de resultados parciales o finales
de investigaciones. El texto del Artículo científico se
divide en secciones que llevan estos
encabezamientos:
 Introducción
Materiales y Métodos
Resultados
Discusión
Conclusiones e implicaciones
Literatura citada
En los artículos largos puede ser necesario agregar
subtítulos dentro de estas divisiones a fin de hacer
más claro el contenido, sobre todo en las secciones de
Resultados y de Discusión, las cuales también pueden
presentarse como una sola sección.
b) Notas de investigación. Consisten en
modificaciones a técnicas, informes de casos clínicos
de interés especial, preliminares de trabajos o
investigaciones limitadas, descripción de nuevas
variedades de pastos; así como resultados de
investigación que a juicio de los editores deban así ser
publicados. El texto contendrá la misma información
del método experimental señalado en el inciso a),
pero su redacción será corrida del principio al final del
trabajo; esto no quiere decir que sólo se supriman los
subtítulos, sino que se redacte en forma continua y
coherente.
c) Revisiones bibliográficas. Consisten en el
tratamiento y exposición de un tema o tópico de
relevante actualidad e importancia; su finalidad es la
de resumir, analizar y discutir, así como poner a
disposición del lector información ya publicada sobre
un tema específico. El texto se divide en:
Introducción, y las secciones que correspondan al
desarrollo del tema en cuestión.
11. Agradecimientos y conflicto de interés. Siempre
que corresponda, se deben especificar las
colaboraciones que necesitan ser reconocidas, tales
como a) la ayuda técnica recibida; b) el
agradecimiento por el apoyo financiero y material,
especificando la índole del mismo; c) las relaciones
financieras que pudieran suscitar un conflicto de
intereses. Las personas que colaboraron pueden ser
citadas por su nombre, añadiendo su función o tipo de
colaboración; por ejemplo: “asesor científico”,
“revisión crítica de la propuesta para el estudio”,
“recolección de datos”, etc. Siempre que corresponda,
los autores deberán mencionar si existe algún
conflicto de interés.
12. Literatura citada. Numere las referencias
consecutivamente en el orden en que se mencionan
por primera vez en el texto. Las referencias en el
texto, en los cuadros y en las ilustraciones se deben
identificar mediante números arábigos entre
paréntesis, sin señalar el año de la referencia. Evite
hasta donde sea posible, el tener que mencionar en el
texto el nombre de los autores de las referencias.
Procure abstenerse de utilizar los resúmenes como
referencias; las “observaciones inéditas” y las
“comunicaciones personales” no deben usarse como
VII
referencias, aunque pueden insertarse en el texto
(entre paréntesis).
Reglas básicas para la Literatura citada
Nombre de los autores, con mayúsculas sólo las
iniciales, empezando por el apellido paterno, luego
iniciales del materno y nombre(s). En caso de
apellidos compuestos se debe poner un guión entre
ambos, ejemplo: Elías-Calles E. Entre las iniciales de
un autor no se debe poner ningún signo de
puntuación, ni separación; después de cada autor sólo
se debe poner una coma, incluso después del
penúltimo; después del último autor se debe poner un
punto.
El título del trabajo se debe escribir completo (en su
idioma original) luego el título abreviado de la revista
donde se publicó, sin ningún signo de puntuación;
inmediatamente después el año de la publicación,
luego el número del volumen, seguido del número
(entre paréntesis) de la revista y finalmente el número
de páginas (esto en caso de artículo ordinario de
revista).
Puede incluir en la lista de referencias, los artículos
aceptados aunque todavía no se publiquen; indique la
revista y agregue “en prensa” (entre corchetes).
En el caso de libros de un solo autor (o más de uno,
pero todos responsables del contenido total del libro),
después del o los nombres, se debe indicar el título
del libro, el número de la edición, el país, la casa
editorial y el año.
Cuando se trate del capítulo de un libro de varios
autores, se debe poner el nombre del autor del
capítulo, luego el título del capítulo, después el
nombre de los editores y el título del libro, seguido del
país, la casa editorial, año y las páginas que abarca el
capítulo.
En el caso de tesis, se debe indicar el nombre del
autor, el título del trabajo, luego entre corchetes el
grado (licenciatura, maestría, doctorado), luego el
nombre de la ciudad, estado y en su caso país,
seguidamente el nombre de la Universidad (no el de
la escuela), y finalmente el año.
Emplee el estilo de los ejemplos que aparecen a
continuación, los cuales están parcialmente basados
en el formato que la Biblioteca Nacional de Medicina
de los Estados Unidos usa en el Index Medicus.
Revistas
Artículo ordinario, con volumen y número. (Incluya el
nombre de todos los autores cuando sean seis o
menos; si son siete o más, anote sólo el nombre de
los seis primeros y agregue “et al.”).
I) Basurto GR, Garza FJD. Efecto de la inclusión de grasa
o proteína de escape ruminal en el comportamiento
de toretes Brahman en engorda. Téc Pecu Méx
1998;36(1):35-48.
Sólo número sin indicar volumen.
II) Stephano HA, Gay GM, Ramírez TC. Encephalomielitis,
reproductive failure and corneal opacity (blue eye) in
pigs associated with a paramyxovirus infection. Vet
Rec 1988;(122):6-10.
III) Chupin D, Schuh H. Survey of present status ofthe use
of artificial insemination in developing countries.
World Anim Rev 1993;(74-75):26-35.
No se indica el autor.
IV) Cancer in South Africa [editorial]. S Afr Med J
1994;84:15.
Suplemento de revista.
V) Hall JB, Staigmiller RB, Short RE, Bellows RA, Bartlett
SE. Body composition at puberty in beef heifers as
influenced by nutrition and breed [abstract]. J Anim
Sci 1998;71(Suppl 1):205.
Organización, como autor.
VI) The Cardiac Society of Australia and New Zealand.
Clinical exercise stress testing. Safety and performance
guidelines. Med J Aust 1996;(164):282-284.
En proceso de publicación.
VII) Scifres CJ, Kothmann MM. Differential grazing use of
herbicide treated area by cattle. J Range Manage [in
press] 2000.
Libros y otras monografías
Autor total.
VIII) Steel RGD, Torrie JH. Principles and procedures of
statistics: A biometrical approach. 2nd ed. New
York, USA: McGraw-Hill Book Co.; 1980.
Autor de capítulo.
IX) Roberts SJ. Equine abortion. In: Faulkner LLC editor.
Abortion diseases of cattle. 1rst ed. Springfield,
Illinois, USA: Thomas Books; 1968:158-179.
Memorias de reuniones.
X) Loeza LR, Angeles MAA, Cisneros GF. Alimentación
de cerdos. En: Zúñiga GJL, Cruz BJA editores.
Tercera reunión anual del centro de investigaciones
forestales y agropecuarias del estado de Veracruz.
Veracruz. 1990:51-56.
VIII
XI) Olea PR, Cuarón IJA, Ruiz LFJ, Villagómez AE.
Concentración de insulina plasmática en cerdas
alimentadas con melaza en la dieta durante la
inducción de estro lactacional [resumen]. Reunión
nacional de investigación pecuaria. Querétaro, Qro.
1998:13.
XII) Cunningham EP. Genetic diversity in domestic
animals: strategies for conservation and
development. In: Miller RH et al. editors. Proc XX
Beltsville Symposium: Biotechnology’s role in
genetic improvement of farm animals. USDA.
1996:13.
Tesis.
XIII) Alvarez MJA. Inmunidad humoral en la anaplasmosis
y babesiosis bovinas en becerros mantenidos en una
zona endémica [tesis maestría]. México, DF:
Universidad Nacional Autónoma de México; 1989.
XIV) Cairns RB. Infrared spectroscopic studies of solid
oxigen [doctoral thesis]. Berkeley, California, USA:
University of California; 1965.
Organización como autor.
XV) NRC. National Research Council. The nutrient
requirements of beef cattle. 6th ed. Washington,
DC, USA: National Academy Press; 1984.
XVI) SAGAR. Secretaría de Agricultura, Ganadería y
Desarrollo Rural. Curso de actualización técnica para
la aprobación de médicos veterinarios zootecnistas
responsables de establecimientos destinados al
sacrificio de animales. México. 1996.
XVII) AOAC. Oficial methods of analysis. 15th ed.
Arlington, VA, USA: Association of Official Analytical
Chemists. 1990.
XVIII) SAS. SAS/STAT User’s Guide (Release 6.03). Cary
NC, USA: SAS Inst. Inc. 1988.
XIX) SAS. SAS User´s Guide: Statistics (version 5 ed.).
Cary NC, USA: SAS Inst. Inc. 1985.
Publicaciones electrónicas
XX) Jun Y, Ellis M. Effect of group size and feeder type
on growth performance and feeding patterns in
growing pigs. J Anim Sci 2001;79:803-813.
http://jas.fass.org/cgi/reprint/79/4/803.pdf.
Accessed Jul 30, 2003.
XXI) Villalobos GC, González VE, Ortega SJA. Técnicas
para estimar la degradación de proteína y materia
orgánica en el rumen y su importancia en rumiantes
en pastoreo. Téc Pecu Méx 2000;38(2): 119-134.
http://www.tecnicapecuaria.org/trabajos/20021217
5725.pdf. Consultado 30 Ago, 2003.
XXII) Sanh MV, Wiktorsson H, Ly LV. Effect of feeding ha hectárea (s)
level on milk production, body weight change, feed h hora (s)
conversion and postpartum oestrus of crossbred i.m. intramuscular (mente)
lactating cows in tropical conditions. Livest Prod Sci i.v. intravenosa (mente)
2002;27(2-3):331-338. http://www.sciencedirect. J joule (s)
com/science/journal/03016226. Accessed Sep 12, kg kilogramo (s)
2003.
km kilómetro (s)
13. Cuadros, Gráficas e Ilustraciones. Es preferible L litro (s)
que sean pocos, concisos, contando con los datos log logaritmo decimal
necesarios para que sean autosuficientes, que se Mcal megacaloría (s)
entiendan por sí mismos sin necesidad de leer el texto. MJ megajoule (s)
Para las notas al pie se deberán utilizar los símbolos
m metro (s)
convencionales.
msnm metros sobre el nivel del mar
14 Versión final. Es el documento en el cual los autores µg microgramo (s)
ya integraron las correcciones y modificaciones µl microlitro (s)
indicadas por el Comité Revisor. Los trabajos deberán
µm micrómetro (s)(micra(s))
ser elaborados con Microsoft Word. Las fotografías e
imágenes deberán estar en formato jpg (o mg miligramo (s)
compatible) con al menos 300 dpi de resolución. ml mililitro (s)
Tanto las fotografías, imágenes, gráficas, cuadros o mm milímetro (s)
tablas deberán incluirse en el mismo archivo del texto. min minuto (s)
Los cuadros no deberán contener ninguna línea ng nanogramo (s)Pprobabilidad (estadística)
vertical, y las horizontales solamente las que delimitan p página
los encabezados de columna, y la línea al final del PC proteína cruda
cuadro.
PCR reacción en cadena de la polimerasa
15. Una vez recibida la versión final, ésta se mandará para pp páginas
su traducción al idioma inglés o español, según ppm partes por millón
corresponda. Si los autores lo consideran conveniente % por ciento (con número)
podrán enviar su manuscrito final en ambos idiomas.
rpm revoluciones por minuto
16. Tesis. Se publicarán como Artículo o Nota de seg segundo (s)
Investigación, siempre y cuando se ajusten a las t tonelada (s)
normas de esta revista. TND total de nutrientes digestibles
17. Los trabajos no aceptados para su publicación se UA unidad animal
regresarán al autor, con un anexo en el que se UI unidades internacionales
explicarán los motivos por los que se rechaza o las vs versus
modificaciones que deberán hacerse para ser xg gravedades
reevaluados.
Cualquier otra abreviatura se pondrá entre paréntesis
18. Abreviaturas de uso frecuente: inmediatamente después de la(s) palabra(s)
cal caloría (s) completa(s).
cm centímetro (s) 19. Los nombres científicos y otras locuciones latinas se
°C grado centígrado (s) deben escribir en cursivas.
DL50 dosis letal 50%
g gramo (s)

IX

INSTRUCTIONS FOR AUTHORS
Revista Mexicana de Ciencias Pecuarias is a scientific
journal published in a bilingual format (Spanish and
English) which carries three types of papers: Research
Articles, Technical Notes, and Reviews. Authors interested
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mentioned directives which are based on those set down
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6. Manuscripts of all three type of articles published in
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X
Updated: March, 2020
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c. Accepted articles, even if still not published, can
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d. In the case of a single author’s book (or more
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the title of the book should be indicated after the
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XI
e. When a reference is made of a chapter of book
written by several authors; the name of the author(s)
of the chapter should be quoted, followed by the title
of the chapter, the editors and the title of the book,
the country, the printing house, the year, and the
initial and final pages.
f. In the case of a thesis, references should be
made of the author’s name, the title of the research,
the degree obtained, followed by the name of the City,
State, and Country, the University (not the school),
and finally the year.
Examples
The style of the following examples, which are partly
based on the format the National Library of Medicine
of the United States employs in its Index Medicus,
should be taken as a model.
Journals
Standard journal article (List the first six authors
followed by et al.)
I) Basurto GR, Garza FJD. Efecto de la inclusión de grasa
o proteína de escape ruminal en el comportamiento
de toretes Brahman en engorda. Téc Pecu Méx
1998;36(1):35-48.
Issue with no volume
II) Stephano HA, Gay GM, Ramírez TC. Encephalomielitis,
reproductive failure and corneal opacity (blue eye) in
pigs associated with a paramyxovirus infection. Vet
Rec 1988;(122):6-10.
III) Chupin D, Schuh H. Survey of present status of the
use of artificial insemination in developing countries.
World Anim Rev 1993;(74-75):26-35.
No author given
IV) Cancer in South Africa [editorial]. S Afr Med J
1994;84:15.
Journal supplement
V) Hall JB, Staigmiller RB, Short RE, Bellows RA, Bartlett
SE. Body composition at puberty in beef heifers as
influenced by nutrition and breed [abstract]. J Anim
Sci 1998;71(Suppl 1):205.
Organization, as author
VI) The Cardiac Society of Australia and New Zealand.
Clinical exercise stress testing. Safety and
performance guidelines. Med J Aust 1996;(164):282-
284.
In press
VII) Scifres CJ, Kothmann MM. Differential grazing use of
herbicide-treated area by cattle. J Range Manage [in
press] 2000.
Books and other monographs
Author(s)
VIII) Steel RGD, Torrie JH. Principles and procedures of
statistics: A biometrical approach. 2nd ed. New
York, USA: McGraw-Hill Book Co.; 1980.
Chapter in a book
IX) Roberts SJ. Equine abortion. In: Faulkner LLC editor.
Abortion diseases of cattle. 1rst ed. Springfield,
Illinois, USA: Thomas Books; 1968:158-179.
Conference paper
X) Loeza LR, Angeles MAA, Cisneros GF. Alimentación
de cerdos. En: Zúñiga GJL, Cruz BJA editores.
Tercera reunión anual del centro de investigaciones
forestales y agropecuarias del estado de Veracruz.
Veracruz. 1990:51-56.
XI) Olea PR, Cuarón IJA, Ruiz LFJ, Villagómez AE.
Concentración de insulina plasmática en cerdas
alimentadas con melaza en la dieta durante la
inducción de estro lactacional [resumen]. Reunión
nacional de investigación pecuaria. Querétaro, Qro.
1998:13.
XII) Cunningham EP. Genetic diversity in domestic
animals: strategies for conservation and
development. In: Miller RH et al. editors. Proc XX
Beltsville Symposium: Biotechnology’s role in
genetic improvement of farm animals. USDA.
1996:13.
Thesis
XIII) Alvarez MJA. Inmunidad humoral en la anaplasmosis
y babesiosis bovinas en becerros mantenidos en una
zona endémica [tesis maestría]. México, DF:
Universidad Nacional Autónoma de México; 1989.
XIV) Cairns RB. Infrared spectroscopic studies of solid
oxigen [doctoral thesis]. Berkeley, California, USA:
University of California; 1965.
Organization as author
XV) NRC. National Research Council. The nutrient
requirements of beef cattle. 6th ed. Washington,
DC, USA: National Academy Press; 1984.
XVI) SAGAR. Secretaría de Agricultura, Ganadería y
Desarrollo Rural. Curso de actualización técnica para
la aprobación de médicos veterinarios zootecnistas
responsables de establecimientos destinados al
sacrificio de animales. México. 1996.
XII
XVII) AOAC. Official methods of analysis. 15th ed.
Arlington, VA, USA: Association of Official Analytical
Chemists. 1990.
XVIII) SAS. SAS/STAT User’s Guide (Release 6.03). Cary
NC, USA: SAS Inst. Inc. 1988.
XIX) SAS. SAS User´s Guide: Statistics (version 5 ed.).
Cary NC, USA: SAS Inst. Inc. 1985.
Electronic publications
XX) Jun Y, Ellis M. Effect of group size and feeder type
on growth performance and feeding patterns in
growing pigs. J Anim Sci 2001;79:803-813.
http://jas.fass.org/cgi/reprint/79/4/803.pdf.
Accesed Jul 30, 2003.
XXI) Villalobos GC, González VE, Ortega SJA. Técnicas
para estimar la degradación de proteína y materia
orgánica en el rumen y su importancia en rumiantes
en pastoreo. Téc Pecu Méx 2000;38(2): 119-134.
http://www.tecnicapecuaria.org/trabajos/20021217
5725.pdf. Consultado 30 Jul, 2003.
XXII) Sanh MV, Wiktorsson H, Ly LV. Effect of feeding
level on milk production, body weight change, feed
conversion and postpartum oestrus of crossbred
lactating cows in tropical conditions. Livest Prod Sci
2002;27(2-3):331-338.
http://www.sciencedirect.com/science/journal/030
16226. Accesed Sep 12, 2003.
12. Tables, Graphics and Illustrations. It is preferable
that they should be few, brief and having the
necessary data so they could be understood without
reading the text. Explanatory material should be
placed in footnotes, using conventional symbols.
13. Final version. This is the document in which the
authors have already integrated the corrections and
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works will have to be elaborated with Microsoft Word.
Photographs and images must be in jpg (or
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15. Thesis will be published as a Research Article or as a
Technical Note, according to these guidelines.
16. Manuscripts not accepted for publication will be
returned to the author together with a note explaining
 the cause for rejection, or suggesting changes which ml milliliter (s)
should be made for re-assessment. mm millimeter (s)
min minute (s)
17. List of abbreviations:
ng nanogram (s)
cal calorie (s) P probability (statistic)
cm centimeter (s) p page
°C degree Celsius CP crude protein
DL50 lethal dose 50% PCR polymerase chain reaction
g gram (s) pp pages
ha hectare (s) ppm parts per million
h hour (s) % percent (with number)
i.m. intramuscular (..ly) rpm revolutions per minute
i.v. intravenous (..ly) sec second (s)
J joule (s) t metric ton (s)
kg kilogram (s) TDN total digestible nutrients
km kilometer (s) AU animal unit
L liter (s) IU international units
log decimal logarithm vs versus
Mcal mega calorie (s) xg gravidity
MJ mega joule (s)
The full term for which an abbreviation stands should
m meter (s) precede its first use in the text.
µl micro liter (s)
µm micro meter (s) 18. Scientific names and other Latin terms should be
written in italics.
mg milligram (s)

XIII
 https://doi.org/10.22319/rmcp.v14i3.6175

Article

Prediction of the chemical composition of pig feces and ileal digesta by

near-infrared reflectance spectroscopy (NIRS)

Ricardo Ramos Cruz a

Ricardo Basurto Gutiérrez b

Ericka Ramírez Rodríguez b

Tércia Cesária Reis de Souza c

Gerardo Mariscal Landín b*

a
Universidad Nacional Autónoma de México. Facultad de Estudios Superiores Cuautitlán,
México.
b
Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, CENID-Fisiología
y Mejoramiento Animal, Km 1 Carretera a Colón, 76280 Ajuchitlán, Querétaro, México.
c
Universidad Autónoma de Querétaro. Facultad de Ciencias Naturales, Querétaro, México.

*Corresponding author: mariscal.gerardo@inifap.gob.mx

Abstract:

Near-infrared reflectance spectroscopy (NIRS) allows the estimation of the chemical

composition of various samples (ingredients, animal products, etc.); however, there is little
information on its use with samples of swine ileal digesta (ID) or feces (F). Therefore, the
objective was to develop prediction equations based on the partial least squares method to
predict the chemical composition of the ID and F through NIRS. 110 ID and 202 F samples
from digestibility experiments were used, their spectra were obtained and a multivariate
model was used to develop the prediction method. The variables analyzed in ID were: crude
protein (CP), leucine (Leu), lysine (Lys) and threonine (Thr) and in F: dry matter (DM), CP
and energy (E). The DI values were: PC: R2 0.98, standard error of calibration (SEC) 0.330,
standard error of prediction (SEP) 0.640; Leu: R2 0.95, SEC 0.040, SEP 0.102; Lys: R2 0.93,

488
 Rev Mex Cienc Pecu 2023;14(3):488-504

SEC 0.077, SEP 0.143; Thr: R2 0.67, SEC 0.209, SEP 0.187. F values were: PC: R2 0.98,
SEC 0.95, SEP 1.19; E (kcal/kg): R2 0.94, SEC 60.8, SEP 95.3; MS: R2 0.87, SEC 0.83, SEP
1.15. The results show that the calibration robustness (SD/SEP) was good for PC, 3.34, Leu
2.07 and Lys 2.48 and fair for Thr 1.94, the prediction (RPD) was good for PC 2.11 in ID. In
F the R2 were high for PC 0.98 and E 0.94. The highest robustness was for PC 5.59 and its
prediction was excellent 4.16 and good for E 2.53. It is concluded that NIRS can predict PC
in ID and PC and E in F. In order to improve the estimation of amino acids in ID, the causes
affecting the robustness of the calibrations should be explored.

Keywords: Prediction, Protein, Energy, Ileal, Fecal.

Received: 16/03/2022

Accepted: 21/02/2023

Introduction

Pig farms are increasingly obliged to improve their production efficiency as they are
estimated to emit 9 % of total greenhouse gas emissions from livestock production; 27.4 %
of this total is contributed by excreta, which is composed of 70.1 % methane and 29.9 %
nitrous oxide(1). Nitrous oxide can be reduced by improving feed efficiency and thus
decreasing nitrogen excretion(2). In this respect, the greatest mitigation potential is found in
the semi-technified system, which is the result of an improvement in herd efficiency(1),
although such a decrease in nitrogen excretion can be brought about in any system by
improving the accuracy of feed formulation(3).

One tool to achieve this objective is the determination of the digestibility of the raw materials
and diets used to feed pigs, especially in a production system such as the Mexican one, which
depends on a wide variety of raw materials, unlike the corn-soybean meal system used in
some countries such as the United States and Brazil. However, developing these studies is
time-consuming and costly, mainly because of the laboratory analyses that must be
performed. The National Institute for Research on Forestry, Agriculture, and Livestock
(Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, INIFAP) has
conducted several studies on ileal digestibility of amino acids and total protein and energy(4).
Therefore, the ileal digesta and stool samples from these studies were used to generate the
calibration model based on the partial least squares method to estimate the chemical
composition of the ileal and stool contents in a near-infrared equipment.

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The near infrared belongs to the electromagnetic spectrum, which is divided into regions
according to the length of the emitted waves, the near infrared region (NIRS) is comprised
between the wavelengths of 780 to 2,500 nm, and is characterized because its radiation does
not modify the structure of matter, but produces vibration of its atoms, hence its wide use in
industry and animal production(5,6). H, C, N, and O atoms form the functional groups C-H,
N-H and O-H, which absorb part of the emitted energy and reflect part of it, creating
absorption bands. The proportion and quantity of these functional groups found in the sample
make its spectrum unique(7); so NIRS can accurately predict the chemical composition of
scanned samples. However, before using it as a tool to predict the chemical composition of
a sample, it is necessary to obtain prediction models for each type of sample and analyte to
be analyzed. This process consists in associating the spectral information with the values
obtained in the laboratory(8). Therefore, the objective of the present work was to obtain
prediction models in a NIRS apparatus to estimate the crude protein, lysine, threonine, and
leucine content in ileal digesta, as well as the dry matter, protein, and energy content in pig
feces.

Material and methods

The study was carried out at the National Center for Research on Animal Physiology and
Improvement (Centro Nacional de Investigación en Fisiología y Mejoramiento Animal,
CENID-Fisiología), in Ajuchitlán Querétaro, Mexico. Ileal digesta and stool samples were
obtained from previously performed experiments(9-15), ileal and fecal digestibility of dry
matter (DM), energy (E), and crude protein (CP), and ileal digestibility of amino acids were
determined (AA). In all experiments, the guidelines of the CIOMS(16) and the Mexican
Official Standard for the production, care, and use of laboratory animals were respected(17).

Animals, cages, and sample collection

All pigs were housed in individual metabolic cages equipped with feeders, nipple water
dispensers, and feces collection trays, all feed was in the form of meal and is described in the
references cited; pigs were fed at 2.5 times their maintenance DE requirements(18). The
facility has a temperature control system which was maintained at 19 ± 2 °C. Ileal digesta
samples were freeze-dried, and feces samples were dried in a forced-air oven at 55 °C for 48
h. Subsequently, freeze-dried digesta samples were ground through a 0.5 mm mesh, and dried
stool samples, were through a 1 mm mesh in a laboratory mill (Arthur H. Thomas Co.
Philadelphia, PA).

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Laboratory analysis

Dry matter and crude protein analyses were performed according to AOAC official methods
934.01 and 976.05(19). The digestibility markers used in the experiments were determined for
chromium oxide according to Fenton(20) and titanium oxide according to Myers(21). The AA
were determined by AOAC(19) method No. 994.12, which consists of hydrolyzing the
samples at 110 °C for 24 h in 6 mol/L HCl. Amino acid analysis was performed according to
Henderson(22) or Csapó(23). Energy analyses were carried out in an adiabatic calorimetric
pump (1281, Parr, Moline, IL).

Procedure for obtaining samples in NIRS equipment

The samples were placed in a quartz cup and scanned with a Nicolet 6700 FT-IR
spectrophotometer (Thermo Fisher Scientific, Inc); the spectra were generated with the
OMNIC™ software. The spectra were obtained in the near infrared (NIR) region (1000-2500
nm); the data absorbance was expressed as the logarithm of the reciprocal of R "(log 1/R)",
where R= reflectance. The spectra thus generated were fed with the values obtained in the
laboratory. The multivariate statistical model used was partial least squares, the first or
second derivative, and the Savitzky-Golay filters with the TQ Analyst v8™ software. The
variables entered in TQ Analyst v8 were: dry matter, protein, energy, and amino acid content.
Calibration was performed with 2/3 of the samples, and validation, with 1/3, either for stool
sample or ileal content. The selection of the best predictive model was based on the
minimization of the standard error of prediction (SEP) and of the standard error of cross-
calibration (SECV), and the maximization of the coefficient of determination (R2). In order
to evaluate The predictive power of the calibration model was assessed considering the index
known as RPD, i.e., the predictive deviation ratio. The predictive power of the calibration is
considered as follows: if the RPD is less than 1.5, the calibration is not useful; if the RPD is
between 1.5 and 2.0, the calibration may distinguish between low and high values; if the RPD
is between 2.0 and 2.5, the calibration has possibilities to make quantitative predictions; if
the RPD is between 2.5 to 3.0, the calibration prediction is good, and if RPD is greater than
3.0, the calibration leads to excellent predictions (Williams, 2003) quoted by Saeys(24).
Samples with differences of over 2.5 standard deviations were considered "outliers" and
eliminated from the study.

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Scanning of samples

Ileal digesta: A total of 119 ileal digesta samples were used. The mathematical analysis
utilized was partial least square (PMS). The mathematical treatment was 1,4,13,6 (the
numbers represent the degree of the derivative, the width of the interval over which the
derivative is calculated, the smoothness level, and the order level of the polynomial).

Feces: A total of 222 samples were utilized. The statistical model used was the partial least
square (PMS), and the mathematical treatment utilized was 1,4,11,5 (see explanation above).

Graphs. The graphs were made using the "ggplot2"(25) and "patchwork"(26) packages of R
software(27).

Results

Spectra

Representative spectra of ileal digesta and feces samples are shown in Figure 1.

Figure 1: Representative spectra of ileal digesta and feces samples

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Calibration for ileal digesta analysis

A total of 119 samples were used, of which 9 were eliminated because they were considered
to be "outliers" according to the aforementioned criteria. Thus, the models were developed
over 110 samples.

Protein. The mean and standard deviation (SD) for protein were 15.57 ± 2.15, ranging from
11.1 to 20.8 (Table 1). The coefficient of determination R2 was 0.98, the calibration standard
error (SEC), cross-validation standard error (SECV) and prediction standard error (SEP)
were 0.330, 1.020 and 0.640, respectively. Its RPD was 2.11 and the SD/SEP ratio was 3.34
(Table 2, Figure 2A).

Table 1: Statistics of the analyzed nutrients in ileal digestaa

Item Protein Leucine Lysine Threonine
N 110 110 110 110
Mean 15.57 0.85 0.58 0.71
SDb 2.15 0.21 0.29 0.36
minimum 11.1 0.33 0.17 0.32
maximum 20.8 1.31 1.82 2.13
a
The values represent the percentage content of the nutrient in the dry matter, except for energy, which is
expressed in Kcal/kg.
b
SD = standard deviation.

Table 2: Results of analyte calibrations in the ileal digesta

Calibration First Second
Parameter Protein Leucine Lysine Threonine Lysine
R2a 0.98 0.95 0.93 0.67 0.92
SECb 0.330 0.040 0.077 0.209 0.070
SECVc 1.020 0.123 0.217 0.230 0.137
SEPd 0.640 0.102 0.143 0.187 0.115
RPDe 2.11 1.70 1.31 1.57 2.08
SD/SEPf 3.34 2.07 2.00 1.94 2.48
a
Coefficient of determination; bStandard error of calibration; cStandard error of cross-calibration; dStandard
error of prediction; eRatio of the standard deviation to the standard error of the cross-calibration; fRatio of the
standard deviation to the SEP (SD/SEP).

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Figure 2: Relationship between the predicted values by NIRS and those determined in the
laboratory for ileal digesta

Table 2 reports the values of the equations

Leucine. The mean and the standard deviation were 0.85 ± 0.21, with a range of 0.33 to 1.31
(Table 1). The coefficient of determination R2 was 0.95. The values for SEC were 0.040; for
SECV, 0.123, and for SEP, 0.102. The results obtained for RPD were 1.70, and for the
SD/SEP ratio, 2.07 (Table 2, Figure 2B).

Lysine. The mean and the standard deviation were 0.58 ± 0.29, with a range of 0.17 to 1.82.
The coefficient of determination R2 was 0.93. The values were 0.077 for SEC, 0.217 for
SECV, 0.143 for SEP, and 1.31 for RPD, and the SD/SEP ratio was 2.00.

In order to improve the lysine prediction model, a second calibration was performed for
lysine only. The SD/SEP and RPD statistics improved, and there was a minimal decrease
(one percentage point) in R2. The mean standard deviation, and the minimum and maximum
values were the same as described in the previous paragraph; therefore, the coefficient of
determination (R2) was 0.92. The results were 0.070 for SEC, 0.137 for SECV, and 0.115 for
SEP. The result obtained for RPD was 2.08, and 2.48 for SD/SEP. (Table 2, Figure 2C).

Threonine. The mean and standard deviation for threonine were 0.71 ± 0.36, with a range of
0.32 to 2.13 (Table 1). The coefficient of determination R2 was 0.67. The values of SEC,
SECV and SEP were 0.209, 0.230 and 0.187, respectively. Its RPD was 1.57, and the SD/SEP
ratio, 1.94 (Table 2, Figure 2D).

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Calibration for fecal analysis

Of the 222 samples, 20 considered "outliers" were eliminated. Table 3 shows summary
statistics generated by NIRS.

Table 3: Statistics of the analyzed nutrients in the fecesa

Item Protein Energy Dry matter
N 202 202 202
Mean 20.05 4444 93.43
SD 6.65 269 2.48
Minimum 4.50 3684 85.83
Maximum 29.20 4966 97.50
a
The values represent the percentage content of the nutrient in the dry matter, except for energy, which is
expressed in Kcal/kg.
b
SD= Standard deviation.

Protein. Its mean and standard deviation were 20.05 ± 6.65, respectively, with a range of 4.5
to 29.2 (Table 3). Its coefficient of determination R2 was 0.98. The value of SEC was 0.950;
that of SECV was 1.600, and that of SEP was 1.190. The statistics for RPD and SD/SEP were
above 4, i.e., of 4.16 and 5.59, respectively (Table 4, Figure 3A).

Table 4: Results of analyte calibrations in the feces

Parameter Protein Energy Dry matter

R2a 0.98 0.94 0.87
SECb 0.950 60.8 0.830
SECVc 1.600 106.3 1.390
SEPd 1.190 95.3 1.150
RPDe 4.16 2.53 1.78
SD/SEPf 5.59 2.82 2.16
a
Coefficient of determination; bStandard error of calibration; cStandard error of cross-calibration; dStandard
error of prediction; eRatio of standard deviation to the standard error or cross-calibration; fRatio of the
standard deviation to the SEP (SD/SEP).

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Figure 3: Relationship between the predicted values by the NIRS and those determined in
the Laboratory for the feces

Table 4 reports the values of the equations.

Energy. The mean and standard deviation were 4,444 ± 269, ranging from 3,684 to 4,966
(Table 3). The coefficient of determination R2 was 0.94. The results for errors were 60.8 for
SEC, 106.3 for SECV, and 95.3 for SEP. The results obtained for RPD were 2.53, and 2.82
for SD/SEP (Table 4, Figure 3B).

Dry matter. The mean and standard deviation were 93.43 ± 2.48, with a range of 85.83 to
97.50 (Table 3). The coefficient of determination R2 was 0.87; the values of SEC, SECV,
and SEP were 0.830, 1.390, and 1.150, respectively. Its RPD was 1.78, and the SD/SEP ratio
was 2.16 (Table 4, Figure 3C).

Discussion

The basis of the spectroscopy technique is based on the interaction of light with the
molecular composition of matter; in the near infrared (wavelengths between 800 and 2,500
nanometers), energy causes the bonds to excite and vibrate in two ways: shortening (high
energy) or bending (low energy). This can be understood as follows: molecules are a set of
atoms joined by bonds, and each bond vibrates at a certain frequency that depends on the
chemical group involved in it; therefore, the energy of an incident light ray will be absorbed

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when its frequency is identical to the natural frequency of the intermolecular bond (7). Thus,
the arrangement of chemical bonds in a sample makes the spectra unique (28). The organic
bonds (C-H, N-H, O-H) react at different wavelengths and react at different wavelengths
(C-H, N-H, O-H): O-H bonds react in the region between 1,450 nm and 1,900 nm; N-H
bonds react in the region between 2,080 to 2,220 nm and 1,560 to 1,670 nm, and C-H bonds
react in the region between 1,100, 1,600, 1,700-1,800, 2,000, and 2,200-2,400 nm(7,29,30).

Protein and aminoacids

This is the first work in which the NIRS technology was used to predict protein and amino
acid content in ileal digesta in pigs. The results obtained show that, based on the R2 statistic
= 0.97 and the RPD value of 2.11, the calibration for crude protein is acceptable for predicting
ileal digesta composition. These values are in agreement with those indicated by Saeys (24),
who report that the calibration to estimate pig excreta N had a R2= 0.89 and an RPD above
3.0, suggesting that NIRS can predict crude protein content. According to the RPD statistic
(1.70, 1.31, and 1.57), the lysine and leucine concentration prediction is unreliable, but the
calibration obtained can be used to estimate their concentration in the ileal digesta. However,
in the case of threonine, the results were not favorable, as the calibration and prediction
values obtained and the RPD (R2=0.67, SEP=1.87, and RPD=1.57) suggest that this is a
poorly performing calibration and therefore it is not recommended. The low reliability of
threonine prediction may be due to its richness in endogenous ileal protein losses, given that
mucin is rich in threonine and mucin secretion is modulated by several dietary factors such
as fiber and antinutritional factors(31,32). Thus, the concentration of threonine is more variable
than that of the amino acids lysine and leucine; this argument is supported by the ability of
NIRS to predict amino acid content in wheat (213 samples), barley (185 samples), and corn
(258 samples)(33), since the variability in amino acid content in these samples is lower than
in ileal digesta. The prediction was improved in the (second) calibration, in which only lysine
was included; this was attributed to the fact that the correlations obtained were only for that
amino acid ⸻a similar situation to that reported by Owens et al(34), who predicted more than
20 wheat traits and obtained good predictions for only a few of them, including protein. The
results of the present work for predicting stool protein content were superior to or equal to
those obtained in previous studies(35-38).

The range of crude protein values in pig feces (45 to 292 g/kg) is higher than that reported in
previous works(35,36,38), due to the diets used in them; for example, in the study by
Bastianelli(36), the samples came from a single experimental diet, whereas in the present study
they came from several experiments and, consequently, from different diets. However, the
greater variation in the data of the present work makes the resulting calibration more robust;

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this can be seen in the R2, which is higher than those reported in those other works (0.98 vs
0.84, 0.88, and 0.89)(35,36,38) and indicates that the calibration was excellent. According to
Saeys(24) calibration can be used to predict the crude protein content of the stools. This is
reflected in the results of the present study, where the error for prediction was 11.9 g/kg, a
lower value than those reported in other studies. In addition, the calibration is considered to
have an RPD statistic value of 4.15, which is higher than the value of 3 recommended by
Saeys(24), which indicates that the prediction is accurate and reliable.

Gross energy

The energy prediction is achieved thanks to the organic C-H bond absorption bands present
in the scanned samples and corresponds to those bonds present in the carbohydrates, lipids,
and protein of the food. Carbohydrates present in the feed constitute the main fraction of the
pigs’ diet, and are classified into sugars (mono and disaccharides), oligosaccharides, and
polysaccharides (starch and non-starch polysaccharides (NSP)). Most of the disaccharides
and starch are digested and absorbed in the small intestine as shown by the work of Reis et
al(10); while the pig's digestive enzymes do not hydrolyze NSP, and therefore their
degradation is carried out by the microflora of the cecum and colon through fermentation(39).

The dietary protein source varies in its amino acid content according to the pig's stage of
production. For example, piglets consume mainly animal sources such as whey proteins, fish,
animal plasma, etc., which are highly digestible. As the pig’s digestive system matures,
vegetable proteins (mainly from soybean meal and other oilseeds) increase until only
vegetable proteins are used, their digestibility being more variable than that of carbohydrates
or fats(40). In ileal digesta samples, these bonds correspond mostly to the undigested bonds of
the consumed feed and to the endogenous losses contributed by the animal's organism(41).
Substrates not absorbed in the small intestine reach the large intestine and are fermented by
the microbiota or excreted in the feces(42). In the case of amino acids, they can be used for
microbial protein synthesis or fermented and used as a source of energy(43). Therefore, the C-
H bonds in the feces correspond mainly to the bonds in the microbiota and the fermentation
products produced by it.

The results obtained in the calibration to predict energy were excellent; the R 2 was 0.94,
which indicates an excellent calibration that can be used in the prediction of the energy
content of feces(24). The results of the present study are consistent with other studies(37,38). In
addition, the robustness of the energy prediction (RPD of 2.52) means that quantitative
predictions are possible.

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The results regarding the predictive ability of stool protein and energy content are similar to
those of Cruz-Conesa(44), whose objective was to develop predictive models to estimate the
chemical composition of the feces of three different poultry species: broilers, layers, and
turkeys. These authors(44) reported that their calibrations accounted for over 93 % of the
variation, and the RPDs were greater than 3.7, concluding that NIRS is capable of estimating
feces' protein and energy content.

Dry matter

The chemical composition of the samples is affected by the collection site (ileal or fecal),
since, although most of the water is reabsorbed in the small intestine, the large intestine is
where the digesta is dehydrated(45); therefore, the feces had a higher dry matter content.

The moisture content of the samples has a direct impact on the generation of the spectrum,
as water increases the absorption of light producing a higher absorption in the whole
spectrum, on the other hand, the solid particles cause the light to be reflected and thus to
reduce the absorption and generate higher peaks. For example, the region between 1,450 nm
and 1,900 nm corresponds to the O-H chemical bonds present in water; consequently, there
are more pronounced peaks in this region.

The coefficient of determination of the calibration for dry matter R2 was 0.87, which indicates
that the calibration is accurate and reliable for determining the dry matter content in ileal
samples according to Saeys(24), with a calibration error of 8.3 g/kg DM. On the other hand,
the prediction error was 11.5 g/kg, and the RPD was 1.78; this value indicates that a
quantitative approximation of the dry matter in the feces samples is possible(24). The
relatively low prediction values obtained for dry matter compared to protein and energy
values result from the fact that the scanned samples were from experiments that had
previously been analyzed for digestibility, and, therefore, the samples had been stored.
Although the samples were stored correctly, they may have become rehydrated with ambient
moisture. This argument is supported by the work of Garnsworthy et al(46), who evaluated
156 wheat grain samples and compared the dry matter values obtained by NIRS on the
previously dehydrated samples versus the dry matter values obtained on the samples that
were dehydrated at the time of scanning, lower values were observed in the previously
dehydrated samples. Another factor is that the variation of the laboratory analysis of the dry
matter was narrow (2.48 %); this small variation means that, when the samples are hydrated,
they easily fall out of the calibrated range, affecting the results.

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Conclusions and implications

It is concluded that NIRS can be used as a rapid and reliable analytical tool for predicting
crude protein content of ileal digesta and crude protein and energy content of feces. In order
to improve the estimation of amino acid content in ID, possible causes affecting the
robustness of the calibrations should be explored The use of NIRS represents a reduction in
the cost of experiments and less time to generate the results of the chemical content of the
samples.

Acknowledgments

This study was partially funded by the National Institute for Research on Forestry,
Agriculture, and Livestock (Instituto Nacional de Investigaciones Forestales, Agrícolas y
Pecuarias) and the Autonomous University of Querétaro (Universidad Autónoma de
Querétaro).

Literature cited:
1. Gerber PJ, Steinfeld H, Henderson B, Mottet A, Opio C, Dijkman J, et al. Enfrentando
el cambio climático a través de la ganadería – Una evaluación global de las emisiones y
oportunidades de mitigación. Roma: FAO; 2013.

2. Milgen JV, Dourmad JY. Concept and application of ideal protein for pigs. J Anim Sci
Biotechnol 2015;6(1):1-11.

3. Andretta I, Pomar C, Rivest J, Pomar J, Lovatto PA, Neto JR. The impact of feeding
growing-finishing pigs with daily tailored diets using precision feeding techniques on
animal performance, nutrient utilization, and body and carcass composition. J Anim Sci
2014;92:3925–3936.

4. Rentería FJA, Gómez RS, López HLH, Ordaz OG, Anaya EAM, Mejía GCA, et al.
Principales aportes de la investigación del INIFAP a la nutrición porcina en México:
retos y perspectivas. Rev Mex Cienc Pecu 2021;12:79-110.

5. Givens DI, Deaville ER. The current and future role of near infrared reflectance
spectroscopy in animal nutrition: a review. Aust J Agric Res 1999;50:1131–1145.

6. Osborne BG. Near-infrared spectroscopy in food analysis. In: Encyclopedia of analytical

chemistry. John Wiley & Sons, Ltd.; 2006:1-14.

500
 Rev Mex Cienc Pecu 2023;14(3):488-504

7. Bertrand D. La spectroscopie proche infrarouge et ses applications dans les industries

de l'alimentation animale. INRA Prod Anim 2002;15(3):209-219.

8. Shenk JS, Westerhaus MO. The application of near infrared reflectance spectroscopy
(NIRS) to forage analysis. In forage quality, evaluation, and utilization. In: Jr. GCF
editor. Forage quality evaluation, and utilization. 1994:406-449.10.2134/1994.

9. Balderrama-Pérez VA, Gómez-Soto JG, Reis de Souza TC, Ramírez RE, Mariscal-
Landín G. Is the kafirin profile capable of modulating the ileal digestibility of amino
acids in a soybean meal-sorghum diet fed to pigs? Anim Nutr 2019;5:124-129.

10. Reis de Souza TC, Ávila AIE, Ramírez RE, Mariscal-Landín G. Effects of kafirins and
tannins concentrations in sorghum on the ileal digestibility of amino acids and starch,
and on the glucose and plasma urea nitrogen levels in growing pigs. Livest Sci
2019;227:29-36.

11. Aguilera A, Reis de Souza TC, Mariscal-Landín G, Escobar K, Montaño S, Bernal MG.
Standardized ileal digestibility of proteins and amino acids in sesame expeller and soya
bean meal in weaning piglets. J Anim Physiol Anim Nutr 2015;99:728-736.

12. Reis de Souza TC, Escobar García K, Aguilera AB, Ramirez RE, Mariscal-Landín G.
Sesame meal as the first protein source in piglet starter diets and advantages of a phytase:
a digestive study. S Afr J Anim Sci 2017;47:606-615.

13. Mariscal-Landín G, Ramírez RE, Cuarón IJA. Valor nutritivo de subproductos de

cártamo para cerdos en finalización. Rev Mex Cienc Pecu 2017;8:331-340.

14. Mariscal-Landín G, Ramirez RE. Determinación de la digestibilidad de la proteína,

aminoácidos y energía de canola integral en cerdos en crecimiento. Rev Mex Cienc Pecu
2017;8:297-304.

15. Luján OC. Desarrollo tecnológico para el uso del Ronozyme Proact en la producción
porcina [tesis Maestría]. Universidad Nacional Autónoma de México. México: 2014

16. CIOMS. International guiding principles for biomedical research involving animals. In:
Organization WH editor. International guiding principles for biomedical research
involving animals. Council for International Organizations of Medical Sciences ed.
Geneva; 2012.

17. Diario Oficial de la Federación. Especificaciones técnicas para la producción, cuidado

y uso de los animales de laboratorio. Norma Oficial Mexicana NOM-062-ZOO-1999.
Diario Oficial de la Federación 2001(2 de agosto).

18. INRA. L'alimentation des animaux monogastriques: porc, lapin, volailles. Paris, France:
Institut National de la Recherche Agronomique; 1984.

501
 Rev Mex Cienc Pecu 2023;14(3):488-504

19. AOAC. Official Methods of Analysis. 17 th. ed. Arlington, VA. USA: Assoc Offic Anal
Chem; 2000.

20. Fenton TW, Fenton M. An improved procedure for determination of chromic oxide in
feed and feces. Can J Anim Sci 1979;59:631-634.

21. Myers WD, Ludden PA, Nayigihugu V, Hess BW. Technical Note: A procedure for the
preparation and quantitative analysis of samples for titanium dioxide. J Anim Sci
2004;82:179-183.

22. Henderson JH, Ricker RD, Bidlingmeyer BA, Woodward C. Rapid, accurate and
reproducible HPLC analysis of amino acids. Amino acid analysis using Zorbax Eclipse
AAA columns and the Agilent 1100 HPLC. Agilent technologies 2000(Part No.5980-
1193E):10 pag. Agilent technologies home page at: www.agilent.com/chem/supplies.

23. Csapó J, Loki K, Csapó-Kiss Z, Albert C. Separation and determination of the amino
acids by ion exchange column chromatography applying post-column derivatization.
Acta Agr Kapos 2005;9:33-51.

24. Saeys W, Darius P, Ramon H. Potential for on-site analysis of hog manure using a visual
and near infrared diode array reflectance spectrometer. J Near Infrared Spectrosc
2004;12:299–309.

25. Wickham H. Elegant Graphics for data analysis. Nueva York: Springer-Verlag; 2016.
260 Pag. doi:10.1007/978-3-319-24277-4.

26. Patil, I. Visualizations with statistical details: The 'ggstatsplot' approach. J Open Source
Softw 2021;6:3167. doi:10.21105/joss.03167.

27. R Core Team. R: A language and environment for statistical computing. Vienna,
Austria. R Foundation for Statistical Computing. 2021.

28. Tester RF, Karkalas J, Qi X. Starch—composition, fine structure and architecture. J

Cereal Sci 2004;39:151-165.

29. Saeys W, Xing J, de Baerdemaeker J, Ramon H. Comparison of transflectance and

reflectance to analyse hog manures. J Near Infrared Spectroscopy 2005;13:99-107.

30. Wehling RL. Infrared spectroscopy. Food analysis. Boston, MA Springer; 2010:407-
420.

31. Montagne L, Piel C, Lallès JP. Effect of diet on mucin kinetics and composition:
nutrition and health implications. Nutr Rev 2004;62(3):105-114.

502
 Rev Mex Cienc Pecu 2023;14(3):488-504

32. Mariscal-Landín G, Reis de Souza TC, Hernández DAA, Escobar GK. Pérdidas
endógenas de nitrógeno y aminoácidos en cerdos y su aplicación en la estimación de los
coeficientes de digestibilidad ileal de la proteína y aminoácidos de las materias primas.
Téc Pecu Méx 2009;47:371-388.

33. Fontaine J, Schirmer B, Hörr J. Near-infrared reflectance spectroscopy (nirs) enables the
fast and accurate prediction of essential amino acid contents. 2. Results for wheat,
barley, corn, triticale, wheat bran/middlings, rice bran, and sorghum. J Agric Food Chem
2002;50:3902-3911.

34. Owens B, McCann MEE, McCracken KJ, Park RS. Prediction of wheat chemical and
physical characteristics and nutritive value by near-infrared reflectance spectroscopy. Br
Poult Sci 2009;50:103-122.

35. Schiborra A, Bulang M, Berk A, Susenbeth A, Schlecht E. Using faecal near-infrared

spectroscopy (FNIRS) to estimate nutrient digestibility and chemical composition of
diets and faeces of growing pigs. Anim Feed Sci Technol 2015;210:234-242.

36. Bastianelli D, Bonnal L, Jaguelin-Peyraud Y, Noblet J. Predicting feed digestibility from

NIRS analysis of pig faeces. Animal 2015;9:781-786.

37. de la Roza-Delgado B, Modroño SFV, Martínez-Fernández A, Soldado A. Suitability of

faecal near-infrared reflectance spectroscopy (NIRS) predictions. Spanish J Agric Res
2015;13:1-7.

38. Nirea KG, Pérez de Nanclares M, Skugor A, Afseth NK, Meuwissen THE, Hansen JØ,
et al. Assessment of fecal near-infrared spectroscopy to predict feces chemical
composition and apparent total-tract digestibility of nutrients in pigs. J Anim Sci
2018;96:2826–2837.

39. Velayudhan DE, Kim IH, Nyachoti CM. Characterization of dietary energy in swine
feed and feed ingredients: a review of recent research results. Asian Australas J Anim
Sci 2015;28:1-13.

40. Aluthge ND, Van Sambeek DM, Carney-Hinkle EE, Li YS, Fernando SC, Burkey TE.
Board invited review: The pig microbiota and the potential for harnessing the power of
the microbiome to improve growth and health. J Anim Sci 2019;97(9):3741-3757.

41. Stein HH, Sève B, Fuller MF, Moughan PJ, de Lange CFM. Invited review: Amino acid
bioavailability and digestibility in pig feed ingredients: Terminology and application. J
Anim Sci 2007;85:172-180.

42. Jha R, Fouhse JM, Tiwari UP, Li L, Willing BP. Dietary fiber and intestinal health of
monogastric animals. Front Vet Sci 2019;6:48.

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 Rev Mex Cienc Pecu 2023;14(3):488-504

43. Lin R, Liu W, Piao M, Zhu H. A review of the relationship between the gut microbiota
and amino acid metabolism. Amino Acids 2017:1-8.

44. Cruz-Conesa A, Ferré J, Pérez-Vendrell AM, Callao MP, Ruisánchez I. Use of visible-
near infrared spectroscopy to predict nutrient composition of poultry excreta. Anim Feed
Sci Technol 2021;283:115-169.

45. Thiagarajah J, Verkman A. Water transport in the gastrointestinal tract. In: Johnson LR
editor. Physiology of the gastrointestinal tract. Fifth ed. Amsterdam: Academic Press;
2012:1757-1780.

46. Garnsworthy PC, Wiseman J, Fegeros K. Prediction of chemical, nutritive and

agronomic characteristics of wheat by near infrared spectroscopy. J Agric Sci
Cambridge 2000;135:409–417.

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 https://doi.org/10.22319/rmcp.v14i3.6294

Article

Effect of hesperidin supplementation on blood profile, antioxidant

capacity, intestinal histomorphology and fecal microbial counts in
Japanese quails

Abdullah Özbilgin a*

Mahmut Niyazi Moğulkoç b

Füsun Erhan Bayçumendur c

Nazlı Ercan d

a
Sivas Cumhuriyet University. Veterinary Faculty. Department of Animal Nutrition and
Nutritional Disorders, Sivas, Turkey.
b
Sivas Cumhuriyet University. Veterinary Faculty. Department of Veterinary
Microbiology, Sivas, Turkey.
c
Sivas Cumhuriyet University. Veterinary Faculty. Department of Veterinary Histology
and Embriology, Sivas, Turkey.
d
Sivas Cumhuriyet University. Veterinary Faculty. Department of Veterinary
Biochemistry, Sivas, Turkey.

*Corresponding author: abdullahozbilgin@gmail.com

Abstract:

This study was conducted to determine the effects of hesperidin, a flavonoid added to
quail diets, on blood serum, enzymes in tissues, intestinal histomorphology and fecal
microflora. In the study, first treatment [(control) (0g hesperidin/kg feed)], second
treatment [(HES1) (1 g hesperidin/kg feed)], third treatment [(HES2) (2 g hesperidin/kg
feed)] was added to with the basal diet through 35 d. The study was carried out with 3
main groups, 20 quails with 5 sub-repeats in each group and a total of 300 quails. At the
end of the study, blood, liver and thigh muscle tissue and fecal samples were taken.
Alanine transaminase (ALT) and lactate dehydrogenase (LDH) decreased in the HES1
group but increased in the HES2 group compared to the control group (P<0.05). Aspartate
transaminase (AST) increased in the HES1 group compared to the control group and

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decreased in the HES2 group (P<0.05). Amylase, on the other hand, showed a regular
increase in HES1 and HES2 groups to which hesperidin was added to the control group
in hesperidin added groups (P<0.05). The tissue antioxidant GSH, CAT and SOD enzyme
parameters showed a significant increase in the hesperidin added groups compared to the
control group, and this increase was found to be significant compared to the control group
(P<0.05). In intestinal histomorphology, in hesperidin treatment groups increased the
height of villus in the cecum tissue; in colon tissue, it was determined that hesperidin
added groups increased villus height but decreased crypt depth (P<0.05). Consequently,
diets hesperidin with treatments positively is thought to affect the lipid, thigh, liver and
serum antioxidant enzyme levels, intestinal histomorphology and fecal microflora in
quail.

Keywords: Blood, Feces, Flavonoid, Lipid, Tissue.

Received: 25/07/2022

Accepted: 08/03/2023

Introduction

Japanese quail (Coturnix coturnix Japonica) has recently received more attention in the
poultry industry due to increased meat and egg yield, especially in European and Latin
American countries. Moreover, quail has become an important model animal for research
due to its short life cycle and great resistance to many poultry diseases(1,2). In quail
feeding, alternative compounds are used today. Citrus have an additional advantage
compared other sources due to the presence of bioactive compounds (flavonoids), used
as functional components for providing health benefits. Bioflavonoids such as hesperidin
and naringenin are abundant as an inexpensive by-product of citrus cultivation(3).
Flavonoids are secondary metabolites of plant origin called antioxidant heterocyclic
organic compounds divided into 14 different subgroups according to the chemical
structure and positions of the components on rings A, B and C(4). Hesperidin (5, 7, 3’-
trihydroxy-4’-methoxy-flavanone 7-rhamno glucoside), one of the primary flavonoids
found in citrus fruit(5), was formed from hesperidin and rutinose(6,7), and have many
biological properties, including antimicrobial, antioxidant and vascular activities(8). These
compounds prevent or delay the oxidation of structures such as proteins, lipids,
carbohydrates. Flavonoids can be conjugated with glucuronic acid, and O-methylation or
sulfate ester formation may occur to facilitate. This type of biotransformation occurs in
the lower parts of the gastrointestinal tract. Some of the flavonoids that are not absorbed
in the small intestine and other compounds are secreted by bile. They also contribute to
the degradation process in the colon, in which microorganisms disrupt the ring

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structures(9). In general, data on the intestinal absorption of hesperidin are insufficient,

and its fate in the intestine is still unclear(10).

In recent studies, it has been reported that when hesperidin is added to quail rations, it
increases meat quality at the end of fattening, positively affects fatty acids in meat, and
egg quality parameters in layer quails(11,12). Other studies reported that it promotes the
activity of cellular antioxidant enzymes such as Superoxide Dismutase (SOD),
Hemeoxygenase-1 (HO-1) and catalase (CAT)(13-17).

In this study, it was hypothesized that the addition of hesperidin to the quail diet would
have positive effects on blood parameters, antioxidant concentration in tissues, intestinal
histomorphology and fecal microflora.

Material and methods

Chemicals

Hesperidin (molecular formula: C28H34O15, cas no: 520-26-13, purity 91%, Chem-Impex
International Company, USA) was purchased commercially in powder form.

Animals and dietary treatments

This study was carried out with the Ethics Committee Approval number 327/2022 of
Sivas Cumhuriyet University Animal Experiments Local Ethics Committee. Japanese
quails (300 quails, weighing 40-45 g and 1-2 wk old) were obtained from breeders in
Sivas (39°42'34.8"N 37°01'13.0"E). The animals were kept in metal cages (Çimuka,
Türkiye) (height: width: depth) (20*45*90 cm) with light for 21 h and darkness for 3 h at
room temperature (25 ± 2 °C) for 35 d. The study was carried out with three main groups,
20 quails with 5 sub-repeats in each group and a total of 300 quails. The first treatment
(control) (0 g hesperidin/kg feed), second treatment (HES1) (1 g hesperidin/kg feed), third
treatment (HES2) (2 g hesperidin/kg feed) was added to with the basal diet through 35 d.

The diets used in the study were formulated according to the recommendations of NRC(18).
(Table 1). The granular feed and water were given ad libitum. Starting the study, an
adaptation period for the environment and the feed was applied for one week. The dose
of hesperidin was determined as is reported in previous study(3). Quails were divided into
three groups as control and study groups to study parameters such as blood parameters,
antioxidant activity, intestinal histomorphology, and fecal microbial counts.

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Table 1: Ingredients and nutrient composition of quail diet

Diets1
Ingredients (%) C HES1 HES2
Corn 44.29 44.29 44.29
Wheat grain 14.80 14.80 14.80
Soybean meal, % 48 37.65 37.65 37.65
Limestone2 0.81 0.71 0.61
Dicalcium phosphate 0.74 0.74 0.74
DL-methionine 0.50 0.50 0.50
L-threonine 0.50 0.50 0.50
Salt 0.42 0.42 0.42
L-lysine, hydrochloride 0.04 0.04 0.04
3
Vitamin-mineral mix 0.25 0.25 0.25
4
Hesperidin 0.00 0.10 0.20
Calculated values
Dry matter, % 89.73 89.63 89.53
Crude protein, % 24.05 24.05 24.05
Ether extract, % 1.85 1.85 1.85
Crude ash, % 5.30 5.30 5.30
Crude cellulose, % 2.89 2.89 2.89
Metabolic energy, kcal/kg 2.900 2.900 2.900
Methionine+Cystine, % 1.27 1.27 1.27
Lysine, % 1.31 1.31 1.31
Threonine, % 1.37 1.37 1.37
Tryptophan, % 0.32 0.32 0.32
Calcium, % 0.61 0.57 0.53
Available phosphorus, % 0.31 0.31 0.31
1
Diets: C – basal diet, HES1 – basal diet with 1 g/kg hesperidin, HES2 – basal diet with 2 g/kg
hesperidin.
2
hesperidin replaced limestone in the same amount in the groups with hesperidin addition.
3
contained per kg: mg: retinol (vitamin A) 3, tocopherol (vitamin E) 30, menadione (vitamin K3) 5,
thiamine (vitamin B1) 1, riboflavin (vitamin B2) 5, pyridoxin (vitamin B6) 3, nicotinic acid 30,
pantothenic acid 10, folic acid 0.8, ascorbic acid (vitamin C) 10, choline chloride 450, Co 0.2, I 0.5, Se
0.3, Fe 25, Mn 120, Cu 10, Zn 100; μg: cholecalciferol (vitamin D3) 62.5, cobalamin (vitamin B12) 20,
biotin 100; 4 obtained from Chem-Impex Int. company, molecule formula (C28H34O15), cas no (520-26-
13), purity grade 91% (Chem-Impex, Wood Dale, IL, USA).

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Slaughter procedures

At the end of the study (35 d) (n=8*3 groups total 24 animals), quails were not fed for 6
h and were slaughtered in a commercial slaughterhouse under hygienic conditions. The
blood of slaughtered quails was taken into blood collection tubes and centrifuged at 3,000
rpm for 10 min. The serum was transferred to 2 ml of microcentrifuge tubes (Eppendorf,
Germany) and kept in a freezer at -80 °C until the analysis were performed. The carcasses
were placed in plastic bags one by one after the internal organs were evacuated and kept
at +4˚C for 24 h. The blood serum, the thigh (Musculus gastrocnemius) and the liver
tissues were kept in a freezer at -80 oC until the analysis were performed.

Blood serum parameters

Tchol (Total cholesterol), BUN (Blood Urea Nitrogen), ALT (Alanine amino
Transferase), AST (Aspartate amino Transferase), GGT (Gamma Glutamyl Transferase),
ALP (Alkaline Phosphatase), TP (Total Protein), TG (Triglyceride), CK (Creatin Kinase),
LDH (Lactate Dehydrogenase), Ca (Calcium), Mg (Magnesium) P (Phosphorus), Urea,
Kreatin, Albumin biochemical values were determined in serum samples using an auto-
analyzer device (Mindray BS200, China).

Tissue parameters

Liver, and thigh tissues were homogenized. Subsequently, GSH levels were determined
using the ELABSCIENCE (E-BC-K030-M); SOD levels were detected using the
ELABSCIENCE (E-BC-K022-M) CAT levels were determined using the
ELABSCIENCE (E-BC-K031-M) MDA (Malondialdehyde) was measured using the
ELABSCIENCE (E-BC-K298-M) LPO levels were detected using the ELABSCIENCE
(E-BC-K176-M) corresponding to the absorbances of the serum and tissue samples
calculated.

Intestinal histomorphology

Ileum, cecum and colon samples were taken into tissue containers for histological
preparation in a 10% buffered formula and washed in tap water for 48 h; 5 µm thick
sections were taken from the tissues, followed by the histological tissue follow-up method
that was applied routinely, were blocked in the paraffin wax. Hematoxylin - eosin staining

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was performed on sections to determine the overall histological structures and perform
histometric measurements(19). The stained preparations were examined under a research
microscope (Zeiss Primo Star, Germany) and their photos were taken. Villus height, villus
width and crypt depth of the 10 pieces obtained from different parts of three sections
belonging to each animal were measured using the ImageJ software. The villus height
was measured at the vertical distance from the villus peaks to the starting point of the
crypts, while the villus widths were obtained from measurements taken at the medium
height of the villus. The depth of the crypt was calculated as the vertical distance from
the villus-crypt junction to the lower boundary of the crypt(20).

Fecal microbial counts

One gram of fecal content from each quail was aseptically collected and homogenized
with 9 mL of 0.1% peptone water. Serial 10-fold dilutions were made in sterile peptone
water from 10–1 to 10–6 and 0.1 mL from last three dilutions were plated in duplicate onto
respective selective medias.

Escherichia coli counts were performed on Tryptone Bile X-Glucuronide (TBX) agar and
plates were incubated for 24 h at 37 °C. Enterococci were cultured on Slanetz Bartley
agar (SB, Oxoid CM377, England) and enumerated after 24-48 h of incubation at 37 ºC.
Enterobacteriaceae and coliforms were grown on violet red bile glucose agar (VRBG,
Oxoid CM485, England) and violet red bile agar (VRB, Oxoid CM107, England)
respectively, using the pour plate technique and enumerated after 24-48 h of incubation
at 37 ºC. Tryptose Sulfite Cycloserine Agar (TSC Agar) Base (Merck 1.11972, Germany)
was utilized for the Clostridium count. The plates were incubated for 24 h at 45 °C under
anaerobic conditions, anaerobic indicator (Mitsubishi Gas Chemical, America) was
included to monitor the atmospheric condition. Petri dishes (90 mm*15 mm) (Fırat
Plastic, Türkiye) observed 30 to 300 colonies were counted using a colony counter(21).
The microbial counts were expressed as log10 CFU per gram of fecal contents.

Statistical analysis

The obtained data were evaluated using the SPSS 20.0 statistical package program. The
data showed normal distribution in all parameters. One-way analysis of variance
(ANOVA) was used and Bonferroni and Tamhane’s T2 multiple comparison test was
used for comparisons between groups (P<0.05).

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Results

Blood serum parameters

Serum alanine transaminase (ALT) and lactate dehydrogenase (LDH) in the HES1 and
the HES2 groups, were no different compared to the control group (P<0.05). Aspartate
transaminase (AST) decreased in the HES2 group compared control group and
HES1(P< 0.05). Amylase, on the other hand, showed an increase in HES2 compared to
the control group (P<0.05) (Table 2). No significant differences were found between the
experiment groups in the parameters of Tchol, TG, GGT, ALP, CK, BUN, Albumin, TP,
Urea, Creatine, Glucose, Ca, Mg and P (P>0.05).

Table 2: The effect of hesperidin added to quail diets on blood serum biochemistry
Control HES1 HES2
Mean± SEM Mean± SEM Mean± SEM P
Tchol, mg/dL 239.82±9.09 262.38±7.22 259.23±5.32 0.12
TG, mg/dL 265.35±24.77 248.40±25.59 247.43±26.77 0.86
ALT, U/L 5.50±0.65ab 4.25±0.48b 6.60±0.25a 0.01*
AST, U/L 237.50±6.89a 243.00±5.51a 194.60±9.66b 0.001*
GGT, U/L 2.00±0.58 2.40±0.40 3.50±0.50 0.14
ALP, U/L 265.73±22.42 279.37±26.89 261.70±26.62 0.89
CK, U/L 2.422±197.57 2.614±211.07 2.650±242.93 0.78
LDH, U/L 711.93±36.69ab 678.36±27.69b 767.81±13.88a 0.03*
Amylase, U/L 243.50±15.50b 302.75±12.15ab 355.50±18.68a 0.01*
BUN, mg/dL 4.00±0.00 4.67±0.33 4.71±0.36 0.26
Albumin, g/dL 1.83±0.11 1.55±0.17 1.83±0.10 0.24
TP, g/dL 6.26±0.71 3.97±0.21 4.74±0.55 0.08
Creatin, mg/dL 0.18±0.02 0.16±0.01 0.19±0.04 0.82
Glucose, mg/dL 56.58±9.66 61.60±5.40 59.20±2.20 0.93
Ca, mg/dL 10.52±0.20 8.87±0.76 9.99±0.10 0.16
Mg, mg/dL 7.58±0.64 6.18±0.50 7.56±0.24 0.09
Tchol= total cholestrol, BUN= blood urea nitrogen, ALT= alanine amino transferase, AST= aspartate
amino transferase, GGT= gamma glutamyl transferase, ALP= alkaline phosphatase, TP= total protein,
TG= triglyceride, CK= creatin kinase, LDH= lactate dehydrogenase, P= phosphorus.
*There is a statistically significant difference between the experimental groups (P<0.05)

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Tissue parameters

GSH level increased in thigh, liver and serum tissue in HES1 group compared to control
group (P<0.05). In thigh tissue, it decreased in the HES2 group compared to the control
group. LPO level increased in thigh and serum tissue in HES1 group compared to control
group. In thigh tissue, it decreased in the HES2 group compared to the control group. and
increased in HES2 groups in serum. In liver tissue, it decreased in HES1 and HES2 groups
compared to the control group (P<0.05). The CAT level in the thigh tissue was decreased
in the hesperidin groups compared to the control group (P<0.05). It increased in liver
tissue (P>0.05). In serum tissue, it decreased in HES1 and increased in HES2 (P<0.05).
MDA level decreased in thigh, liver and serum in HES1 and HES2 groups compared to
control group (P<0.05). On the other hand, while SOD level decreased in hesperidin
groups in thigh tissue compared to the control group (P<0.05); increased in HES1 and
HES2 groups in liver tissue (P<0.05). In the serum tissue, it decreased in the HES1 group
compared to the control group, but increased in the HES2 group (P>0.05) (Table 3).

Table 3: The effect of hesperidin added to the diet on oxidant parameters in thigh, liver
and blood
Control HES1 HES2
Mean± SEM Mean± SEM Mean± SEM P
Thigh 10.79±0.49 12.61±0.16 10.04±0.75 0.06
GSH, µmol/L Liver 6.33±1.01c 12.77±0.65b 19.62±0.65a 0.001*
b a
Serum 51.68±1.34 75.16±1.24 70.88±0.42a 0.01*
Thigh 1.03±0.04a 0.81±0.04b 0.85±0.07b 0.02*
SOD, U/ml
Liver 1.09±0.02 1.11±0.03 1.16±0.01 0.14
Serum 0.37±0.12 0.32±0.13 0.42±0.11 0.84
Thigh 211.62±0.72a 101.55±1.16b 61.56±0.91c 0.001*
CAT, U/ml Liver 76.13±1.31b 88.40±4.18ab 96.69±3.43a 0.01*
Serum 54.30±1.06ab 51.33±1.25b 61.65±1.03a 0.02*
Thigh 758.53±42.62 642.30±43.17 379.71±40.09b 0.02*
a ab

MDA, µmol/L Liver 1.091.16±40.52a 318.98±39.98b 308.31±31.66c 0.001*

Serum 777.69±36.18a 438.24±35.80b 298.32±38.67b 0.01*
Thigh 278.79±1.46a 285.13±1.15a 267.28±2.34b 0.03*
a
LPO, µmol/L Liver 257.09±1.39 241.16±0.78b 249.30±1.29a 0.01*
Serum 241.56±1.56b 279.88±1.42a 284.71±1.52a 0.001*
GSH= reduced glutathione, SOD= superoxide dismutase, CAT= catalase, MDA= malondialdehyde,
LPO= lipid peroxide.
*There is a statistically significant difference between the experimental groups (P<0.05).

Intestinal histomorphology

In terms of histomorphology in the intestine, in ileum tissue, villus height increased in

HES1 and HES2 groups compared to the control group (P<0.05) (Table 4). villus height

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increased in HES1 and HES2 groups compared to the control group in the caecum and
colon tissue (P<0.05). However, crypt depth in colon tissue decreased in HES1 and HES2
groups compared to control group (P<0.05) (Figure 1).

Table 4: The effect of hesperidin added to the diet on villus and crypt depth in the
ileum, caecum and colon
Control HES1 HES2
Mean±SEM Mean±SEM Mean± SEM P
VH 222.42±13.53 234.53±21.69 264.24±22.43 0.33
Ileum VW 67.21±1.97 64.70±5.47 67.10±2.54 0.86
CD 30.50±1.24 31.71±1.82 35.02±3.55 0.41
VH 71.40±3.79b 85.56±4.24a 79.86±1.28ab 0.03*
Caecum VW 41.33±2.47 42.76±1.28 47.81±1.55 0.06
CD 44.45±3.45 38.15±1.78 41.80±1.58 0.22
VH 226.38±18.2b 298.58±22.10ab 312.50±28.75a 0.04*
Colon VW 60.88±2.01b 73.70±3.84a 62.54±3.13ab 0.02*
CD 53.47±4.65a 40.24±2.89b 42.62±2.39ab 0.04*
VH= villus height, VW= villus width, CD= crypt depth.
*There is a statistically significant difference between the experimental groups (P<0.05).

Figure 1: The effect of hesperidin addition to diet on ileum caecum and colon tissues

Control group: ileum (A), caecum (D), colon (G), HES1 group: ileum (B), caecum (E), colon
(H), HES2 group: ileum (C), caecum (F), colon (I) tissue. a: villus hight, b: villus width, c: crypt
depth. Hematoxylin-eosin staining X10.

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Fecal microbial counts

In terms of fecal microflora, Clostridium spp. number decreased in HES1 and HES2
groups compared to the control group (P<0.05) (Table 5). However, there is no significant
difference between the experiment groups in terms of Escherichia coli, Enterococcus
spp., Coliform spp. and Enterobactericeae counts (P>0.05).

Table 5: The effect of dietary hesperidin on bacterial concentration (gram/Log10) in

fecal samples
Control HES1 HES2
Mean± SEM Mean± SEM Mean± SEM p
Escherichia coli 6.51±0.30 6.55±0.27 6.11±0.43 0.61
Enterococcous spp. 6.37±0.34 6.05±0.28 6.77±0.25 0.27
Coliform spp. 6.27±0.36 6.09±0.25 5.44±0.18 0.20
Enterobactericeae 6.43±0.24 5.65±0.26 6.65±0.24 0.88
Clostridium spp. 6.55±0.07a 5.93±0.27a 5.26±0.11b 0.01*
*There is a statistically significant difference between the experimental groups (P<0.05).

Discussion

Serum biochemistry

Dietary antioxidant supplements can improve the physiology and yield characteristics of
animals by changing metabolic processes(17). The current study aimed to investigate the
effects of hesperidin on biochemical parameters and antioxidant indices in quails. Citrus
flavonoids, such as hesperidin and naringin, obtained from citrus fruits and fruit juices
have been associated with lower cholesterol and triglyceride levels in animals in the
previous studies(22,23). The hypocholesterolemic effect of hesperidin has been reported to
be mediated by a decrease in the activity of HMG-Co A reductase(24). In the current study,
both triglyceride and cholesterol levels no decreased in the groups with hesperidin
(P>0.05).

ALT and AST and LDH are enzymes related to liver health, function and it is desirable
that they be low. In rats given 5g/kg of flavonoid daily, it was reported that ALT and AST
concentrations decreased compared to the control group(25). Abdel-Kareem and El-
Sheikh(26) also reported 250, 500 and 1000 mg/kg, Galal et al(27) found that 100 and 150
mg/kg propolis significantly reduced AST and ALT activities in laying hens. In the

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current study, AST enzyme levels decreased in the reason for the decrease in ALT, AST
and LDH with different doses of hesperidin is unknown.

Antioxidant compounds taken from the diet, such as bioflavonoids, can provide some
protection against the early stage of diabetes and the development of complications. It has
been reported that hesperidin significantly reduces glucose in the blood(28). However,
unlike previous studies, the serum glucose concentration increased slightly in the current
study (P>0.05). To interpret the increase in glucose concentration, it is necessary to look
at the plasma insulin concentration.

Tissue oxidant vs antioxidant enzymes

The concentration of SOD enzymes in thigh samples was found to be low, but as high as
expected in liver tissue. In general, this effect of hesperidin on the oxidation parameter
indicates that it can terminate oxidation by adding hydrogen atoms to free radicals. Under
physiological conditions, reactive oxygen species (ROS) production is regulated both
endogenously and by antioxidant supplementation(29,30). The main endogenous
antioxidant enzymes are superoxide dismutase (SOD), catalase (CAT) and glutathione
peroxidase (GPX)(31). SOD converts hydrogen peroxide, which is one of the superoxide
radicals, into oxygen and water under the action of CAT and/or GPX. While SOD and
GPX are formed in many tissues(32), CAT is highly present in liver, kidney tissues and red
blood cells(33). The membranes of immune cells are made of highly polyunsaturated fatty
acids, making them highly sensitive to oxidative stress(34). Therefore, they present high
concentrations of antioxidant enzymes since membrane-related signaling and gene
expression are critical for maintaining immune cell functionality(32). In the current study,
it is believed that the concentrations of thigh and serum tissues and liver and serum tissues
in terms of GSH parameter and CAT parameter, respectively, increase in relation to the
hesperidin supplementation. In another study, it has been reported that the concentration
of hesperidin in animals was lower in terms of SOD activity in rats undergoing a fatigue
test(35). Lien et al(36) and Ting et al(37) reported that the concentration of superoxide
dismutase (SOD) in the blood serum was high with the addition of hesperidin and naringin
(0.5-4.0 g/kg) to broiler rations.

Previous studies reported that a significant decrease in MDA concentration were observed
in broilers consuming rations with grape pulp (1.5, 3 and 6 %) in terms of MDA, which
is one of the oxidant parameters, compared to those consuming a basal diet(38,39). In
another study, when broilers were fed a commercial essential oil mixture (0.05 % of the
diet), a decrease in MDA was observed in the breast (22.4 %) and thigh 62.3 %) muscles
when fed with a commercial essential oil mixture(40). In addition, the inclusion of
hesperidin (0.15 % or 0.3 %) to the rations of broilers has been reported to reduce the
concentration of MDA in the breast muscle(41). In the current study, as in the studies noted
above, it is denser in the liver due to hesperidin, and the MDA value decreased in the

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same way in thigh and serum. In addition, a decrease in LPO, which is an essential
parameter in terms of tissue damage, is expected from the point of view of the current
study due to the dose of hesperidin in thigh and liver tissue. The reason for this is that the
body fat composition of adult quails is rich in unsaturated fatty acids, which is seen in
studies conducted due to the increase in the dose of hesperidin in recent years(11,12).

Intestinal histomorphology

Hesperidin sulfates will be conjugated and absorbed in the duodenum and ileum after oral
administration. Previous studies have shown a small amount of absorption of flavonoid
glycosides in the small intestine(42). The absence of glucuronide detection in intestinal
tissue may be due to the restriction of tissue penetration of glucuronide(43).

Recent studies have reported that flavonoids in a natural form (flavonoid-rich plants) or
the extracts there of was positive effects on immunomodulatory and intestine, including
the increasing villus length in broilers and surface area of the small intestine(44-46).
Kamboh and Zhu(20) found that, among broilers consuming added rations of hesperidin,
the length of the intestinal villus and the width of the villus (21st and 42nd d) increased,
while the depth of the crypt (21 d in the duodenum and ileum, and 42 d in the duodenum)
decreased. In the current study, the height and width of the villus in the ileum, cecum and
colon also increased relatively. Flavonoid compounds that improve intestinal morphology
differences measured in villus height and width have shown that these compounds can
stimulate epithelial cell mitosis. That is because longer or thicker (or both) villus are
associated with active mitosis, and it is believed that the villus continues long-term
absorption without the need for regeneration and a reduced crypt depth. This is because
the crypt is considered a villus factory, and that the large crypt presents rapid tissue
transformation due to inflammation caused by pathogens, toxins, or both(47,48). In
addition, increased villus length/crypt depth ratios with genistein and hesperidin can be
considered an improvement in the digestive system(49). It is that similar results to previous
studies will be obtained in the current study.

Fecal microbial counts

In the current study, a lower concentration was determined in hesperidin groups in terms
of the population of Clostridium spp. in a similar way to the previous studies. Today,
plant extracts commercially used in livestock due to their performance-enhancing and
antimicrobial effects are available in pure or mixed form. It is known that phenolic
compounds and essential oils obtained from plants have antimicrobial effects(16,50). The
intestinal flora contained in the colon promotes the absorption of certain nutrients,

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including polyphenols, from the diet and forms bioactive and absorbable molecules from
dietary compounds(51). Metabolites, consisting of both polyphenols and flora, can
positively effect on health(52). Additionally, numerous benefits of polyphenol metabolites
derived by microflora on the host(53) have been reported.

Flavonoids have the property of low bioavailability and poor absorption. Therefore, they
significantly affect intestinal health(54,55). Phenolic compounds possess bactericidal(53) and
bacteriostatic(54) properties. They minimize the adhesion of pathogenic bacteria (E. coli,
Clostridium spp.) to the intestinal wall and prevent infections in the digestive tract. They
also improve the use of nutrients and animals' performance(55-58).

Zhang et al(59) have reported in their study that polyphenols added in piglet rations did
not have a significant effect of feces E. coli and Clostridia spp. numbers on the 11th and
21st d. Kırkpınar et al(60) have noted a decrease in Clostridium spp. populations in the
ileums of broilers fed with thyme oil, garlic oil and both substances compared to animals
in other groups. Supplementation of cranberry extract (80 mg/kg feed), which is a source
of phenolic acid, anthocyanin, flavonol and flavan-3-ol, to broiler rations has decreased
Enterococcus spp. populations significantly in broilers on the 28th d of the study(61).

Conclusions and implications

As a result, hesperidin supplementation has positive effects in terms of blood parameters,

antioxidant activity, intestinal histomorphology and fecal microflora.

Acknowledgements and conflict of interest

All the authors worked in a coordinated way. The experiment was planned and conducted
by AÖ. Antioxidant parameter analyzes by NE; intestinal histomorphology by FEB,
microflora analysis by AÖ and MNM; The editing part and article writing was done by
AÖ

Literature cited:
1. Ikhlas B, Huda N, Noryati I. Chemical composition and physicochemical properties of
meatballs prepared from mechanically deboned quail meat using various types of
flour. Int J Poult Sci 2011;10:30–37.

2.Purohit AS, Reed C, Mohan A. Development and evaluation of quail breakfast sausage.
LWT 2016;69:447–453.

517
 Rev Mex Cienc Pecu 2023;14(3):505-522

3. Goliomytis M, Kartsonas N, Charismiadou MA, Symeon GK, Simitzis PE, Deligeorgis

SG. The influence of naringin or hesperidin dietary supplementation on broiler meat
quality and oxidative stability. PLoS One 2015;10(10):e0141652.

4.Cushnie TPT, Lamb AJ. Recent advances in understanding the antibacterial properties
of flavonoids. Int J Antimicrob Agents 2011;38:99–107.

5.Cano A, Medina A, Bermej A. Bioactive compounds in different citrus varieties.

Discrimination among cultivars. J Food Compos Anal 2008;21(5):377-381.

6.Cho J. Antioxidant and neuroprotective effects of hesperidin and its aglycone

hesperetin. Arch Pharm Res 2006;29:699-706.

7.Kumar A, Lalitha S, Mishra J. Hesperidin potentiates the neuroprotective effects of

diazepam and gabapentin against pentylenetetrazole-induced convulsions in mice:
Possible behavioral, biochemical and mitochondrial alterations. Indian J Pharmacol
2014;46:309-315.

8.Martini ND, Katerere DRP, Eloff JN. Biological activity of five antibacterial flavonoids
from Combretum erythrophyllum (Combretaceae). J Ethnopharmacol 2004;93:207–
212.

9.Hollman PCH, Katan MB. Dietary flavonoids: intake, health effects and bioavailability.
Food Chem Toxicol 1999;37:937-942.

10.Guo X, Li K, Guo A, Li E. Intestinal absorption and distribution of naringin,

hesperidin, and their metabolites in mice. J Funct Foods 2020;74:104158.

11.Özbilgin A, Kara K, Gümüş R, Tekçe E. Fatty acid compositions and quality of egg
and performance in laying quails fed diet with hesperidin. Trop Anim Health Prod
2021;53:518.

12.Özbilgin A, Kara K, Urcar Gelen S. Effect of hesperidin addition to quail diets on

fattening performance and quality parameters, microbial load, lipid peroxidation and
fatty acid profile of meat. J Anim Feed Sci 2021;30(4):367–378 .

13.Khedr NF. Protective effect of mirtazapine and hesperidin on cyclophosphamide-

induced oxidative damage and infertility in rat ovaries. Exp Biol Med 2015;240(12):
1682–1689.

14.Roohbakhsh A, Parhiz H, Soltani F, Rezzee R, Iranshahi M. Molecular mechanisms

behind the biological effects of hesperidin and hesperetin for the prevention of
cancer and cardiovascular diseases. Life Sci 2015;1(124):64–74 .

15.Parhiz H, Roohbakhsh A, Soltani F, Rezaee R, Iranshahi M. Antioxidant and anti-

inflammatory properties of the citrus flavonoids hesperidin and hesperetin: an
updated review of their molecular mechanisms and experimental models. Phytother
Res 2015;29(3):323–331.

518
 Rev Mex Cienc Pecu 2023;14(3):505-522

16.Purba RAP, Yuangklang C, Paengkoum S, Paengkoum P. Milk fatty acid composition,

rumen microbial population and animal performance in response to diets rich in
linoleic acid supplemented with Piper betle leaves in Saanen goats. Anim Prod Sci
2020. http://dx.doi.org/10.1071/AN20182.

17.Purba RAP, Paengkoum S, Yuangklang C, Paengkoum P, Salem AZM, Liang JB.

Mammary gene expressions and oxidative indicators in ruminal fluid, blood, milk,
and mammary tissue of dairy goats fed a total mixed ration containing piper meal
(Piper betle L.). Ital J Anim Sci 2022;21(1):129-141.

18. National Research Council (NRC). 1994. Nutrient requirements of poultry. 9th
Edition, National Academy Press, Washington DC.

19.Bancroft JD, Gamble M. Theory and practice of histological techniques. 5th ed.
Philadelphia PA. Elsevier Limited, 2002.

20.Kamboh AA, Zhu WY. Individual and combined effects of genistein and hesperidin
on immunity and intestinal morphometry in lipopolysacharide-challenged broiler
chickens. Poult Sci 2014;93:9:2175–2183. https://doi.org/10.3382/ps.2014-03971.

21.Jin LZ, Ho YW, Abdullah N, Jalaludin S. Influence of dried Bacillus subtilis and
lactobacilli cultures on intestinal microflora and performance in broilers. Asian-
Australas J Anim Sci 1996;9:397-404.

22.Selvaraj P, Pugalendi KV. Efficacy of hesperidin on plasma, heart and liver tissue
lipids in rats subjected to isoproterenol-induced cardiotoxicity. Exp Toxicol Pathol
2012;64(5):449-452.

23.Ohtsuki K, Abe A, Mitsuzumi H, Kondo M, Uemura K, Iwasaki Y, Kondo Y. Glucosyl

hesperidin improves serum cholesterol composition and inhibits hypertrophy in
vasculature. J Nutr Sci Vitaminol 2003;49(6):447-450.

24.Lee SJ, Yun YS, Lee IK, Ryoo IJ, Yun BS, Yoo ID. An antioxidant lignan and other
constituents from the root bark of Hibiscus syriacus. Planta Med 1999;65:658–660.

25. Alghazeer R, Elgahmasi S, Elnfati AH, et al. Antioxidant activity and

hepatoprotective potential of flavonoids from Arbutus pavarii against CCl4 induced
hepatic damage. Biotechnol J Int 2017;21(1):1–12.

26. Abdel-Kareem AA, El-Sheikh TM. Impact of supplementing diets with propolis on
productive performance, egg quality traits and some haematological variables of
laying hens. J Anim Physiol Anim Nutr (Berl) 2017;101:441–448.

27. Galal A, Abd El-Motaal AM, Ahmed AMH, Zaki TG. Productive performance and
ımmune response of laying hens as affected by dietary propolis supplementation. Int
J Poult Sci 2008;7:272-278.

519
 Rev Mex Cienc Pecu 2023;14(3):505-522

28.Jung UJ, Lee MK, Jeong KS, Chol MS The hypoglycemic effects of hesperidin and
naringin are partly mediated by hepaticglucose regulating enzymes in C57 BL/KsJ-
db/db Mice. J Nutr 2004;134:2499–2503.

29.Birben E, Sahiner UM, Sackesen C, Erzurum S, Kalayci O. Oxidative stress and

antioxidant defense. World Allergy Organ J 2012;5:9–19.

30. Martemucci G, Costagliola C, Mariano M, D’andrea L, Napolitano P, D’Alessandro

AG. Free radical properties, source and targets, antioxidant consumption and health.
Oxygen 2022;2(2):48-78.

31.Pingitore A, Pace G, Lima P, Mastorci F, Quinones A, Iervasi G, Vassalle C. Exercise

and oxidative stress: Potential effects of antioxidant dietary strategies in sports. Nutr
2015;31:916–922.

32.Griess B, Tom E, Domann F, Teoh-Fitzgerald M. Extracellular superoxide dismutase

and its role in cancer. Free Radic Biol Med 2017;112:464–479.

33.Glorieux C, Calderon PB. Catalase, a remarkable enzyme: Targeting the oldest

antioxidant enzyme to find a new cancer treatment approach. Biol Chem 2017;398:
1095–1108.

34.Knight JA. Review: Free radicals, antioxidants, and the immune system. Ann Clin Lab
Sci 2000;30:145–158.

35.Estruel-Amades S, Massot-Cladera PM, Garcia-Cerdà FJ, Pérez-Cano À, Franch M,

Castell M, Camps-Bossacoma M. Protective effect of hesperidin on the oxidative
stress induced by an exhausting exercise in intensively trained rats. Nutrients
2019;11(4):783.

36.Lien TF, Yeh HS, Su WT. Effect of adding extracted hesperetin, naringenin and pectin
on egg cholesterol, serum traits and antioxidant activity in laying hens. Arch Anim
Nutr 2008;62(1):33-43.

37.Ting S, Yeh HS, Lien TF. Effects of supplemental levels of hesperetin and naringenin
on egg quality, serum traits and antioxidant activity of laying hens. Anim Feed Sci
Technol 2011;163:59-66.

38.Brenes A, Viveros A, Goni I, Centeno C, Sayago-Ayerdy SG, Arija I, Saura-Calixto

F. Effect of grape pomace concentrate and vitamin E on digestibility of polyphenols
and antioxidant activity in chickens. Poult Sci 2008;87:307–316.

39.Goni I, Brenes A, Centeno C, Viveros A, Saura-Calixto F, Rebole A, Arija I, Estevez

R. Effect of dietary grape pomace and vitamin E on growth performance, nutrient
digestibility, and susceptibility to meat lipid oxidation in chickens. Poult Sci 2007;
86:508–516.

520
 Rev Mex Cienc Pecu 2023;14(3):505-522

40.Botsoglou NA, Christaki E, Florou-Paneri P, Giannenas I, Papageorgiou G, Spais AB.

The effect of a mixture of herbal essential oils or α-tocopheryl acetate on
performance parameters and oxidation of body lipid in broilers. S Afr J Anim Sci
2004;34:52–61.

41.Simitzis PE, Symeon GK, Charismiadou MA, Ayoutanti AG, Deligeorgis SG. The
effects of dietary hesperidin supplementation on broiler performance and chicken
meat characteristics. Can J Anim Sci 2011;91:275–282.

42.Escudero-Lopez B, Calani L, Fernandez-Pachon MS, Ortega A, Brighenti F, Crozier

A, Del Rio D. Absorption, metabolism, and excretion of fermented orange juice
(poly)phenols in rats. Biofactors 2014;40(3):327–335.

43.Joshi R, Kulkarni YA, Wairkar S. Pharmacokinetic, pharmacodynamic and

formulations aspects of Naringenin: An update. Life Sci 2018;215;43–56. https://doi.
org/10.1016/j.lfs.2018.10.066.

44.Hanieh H, Gerile C, Narabara K, Gu Z, Abe A, Kondo Y. In vivo immunomodulatory

effects of dietary purple sweet potato after immunization in chicken. Anim Sci J
2010;81:116-121

45.Hassanpour H, Zamani MAK, Yazdani A, Cheraghchi BM. Evaluation of intestinal

morphology and nitric oxide metabolites in broiler chickens supplemented by green
tea. Comp Clin Path 2010;19:43-47.

46.Wallace J, Oleszek W, Franz C, Hahn I, Baser KH, Mathe A, Teichmann K. Dietary

plant bioactives for poultry health and productivity. Br Poult Sci 2010;51:461-487.

47.Giannenas I, Tontis D, Tsalie E, Chronis EF, Doukas D, Kyriazakis I. Influence of

dietary mushroom Agaricus bisporus on intestinal morphology and microflora
composition in broiler chickens. Res Vet Sci 2010;89:78-84.

48.Awad WA, Ghareeb K, Bohm J. Evaluation of the chicory inulin efficacy on

ameliorating the intestinal morphology and modulating the intestinal
electrophysiological properties in broiler chickens. J Anim Physiol Anim Nutr (Berl)
2011;95:65-72.

49.Hu Z, Guo Y. Effects of dietary sodium butyrate supplementation on the intestinal

morphological structure, absorptive function and gut flora in chickens. Anim Feed
Sci Technol 2007;132:240–249.

50.Cross DE, McDevitt RM, Hillman K, Acamovic T. The effect of herbs and their
associated essential oils on performance, dietary digestibility and gut microflora in
chickens from 7 to 28 days of age. Br Poult Sci 2007;48:496-506.

51.Danneskiold-Samsøe NB, Dias de Freitas QBH, Santos R, Bicas JL, Cazarin CBB,
Madsen L, et al. Interplay between food and gut microbiota in health and disease.
Food Res Int 2019;115:23–31.

521
 Rev Mex Cienc Pecu 2023;14(3):505-522

52.Anhê FF, Nachbar RT, Varin TV, Trottier J, Dudonné S, Le Barz M, et al. Treatment
with camu camu (Myrciaria dubia) prevents obesity by altering the gut microbiota
and increasing energy expenditure in diet-induced obese mice. Gut 2019;68:453–
464.

53.Chávez-Carbajal A, Nirmalkar K, Pérez-Lizaur A, Hernández-Quiroz F, Ramírez-Del-

Alto S, García-Mena J, Hernández-Guerrero C. Gut microbiota and predicted
metabolic pathways in a sample of Mexican women affected by obesity and obesity
plus metabolic syndrome. Int J Mol Sci 2019;20:438.

54.Etxeberria U, Fernández - Quintela A, Milagro FI, Aguirre L, Martínez JA, Portill MP.
Impact of polyphenols and polyphenol-rich dietary sources on gut microbiota
composition. J Agric Food Chem 2013;61:9517–9533.

55.Brenes A, Viveros A, Chamorro S, Arija I. Use of polyphenol-rich grape by-products

in monogastric nutrition. A review. Anim Feed Sci Technol 2016;211:1–17.

56.Gordon NC, Png K, Wareham DW. Potent synergy and sustained bactericidal activity
of vancomycin-colistin combination versus multidrug-resistant strains of
Acinetobacter baumannii. Antimicrob Agents Chemother 2010;54:5316-5322.

57.Viveros A, Chamorro S, Pizarro M, Arija I, Centeno C, Brenes A. Effects of dietary

polyphenol-rich grape products on intestinal microflora and gut morphology in
broiler chicks. Poult Sci 2011;90:566–578.

58.Dueñas M, Muñoz - González I, Cueva C, Jiménez – Girón A, Sánchez - Patán F,

Santos - Buelga C, Bartolomé B. A survey of modulation of gut microbiota by dietary
polyphenols. Biomed Res Int 2015;2015:850902.

59.Zhang YS, Li Y, Wang Y, Sun SY, Qu XJ. Naringin, a natural dietary compound,
prevents intestinal tumorigenesis in apc (min/+) mouse model. J Cancer Res Clin
Oncol 2015;178(5):1-13. http://dx.doi.org/10.1007/s00432-015-2097-9.

60.Kırkpınar F, Ünlü KB, Özdemir G. Effects of oregano and garlic essential oils on
performance, carcase, organ and blood characteristics and intestinal microflora of
broilers. Livest Sci 2011;137:219-225.

61.Leusink G, Rempel H, Skura B, Berkyto M, White W, Yang Y, et al. Growth

performance, meat quality, and gut microflora of broiler chickens fed with cranberry
extract. Poult Sci 2010;89(7):1514–1523.

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 https://doi.org/10.22319/rmcp.v14i3.6122

Article

Supplementation of goats with mesquite pod meal in deferred

(Urochloa mosambicenses) grass pasture in the semiarid region

Diego da Hora Souto a

Mara Lúcia Albuquerque Pereira a*

Taiala Cristina de Jesus Pereira a

Herymá Giovane de Oliveira Silva a

Paulo José Presídio Almeida a

Leandro Borges Sousa a

Fernando Oliveira Barreto a

Larisse Borges Sousa a

Karine Pinheiro Oliveira a

Gleidson Giordano Pinto de Carvalho b

a
State University of Southwest Bahia. Postgraduate Program in Animal Science. Campus
of Itapetinga, BR 415, Km 03, 45700-000, Itapetinga, Bahia, Brazil.
b
Federal University of Bahia. School of Veterinary Medicine and Animal Science.
Adhemar de Barros Avenue, 500, Ondina, Salvador, Bahia, Brazil.

∗Corresponding author: marauesb@yahoo.com.br

Abstract:

Supplementation emerges as a simple strategy to improve animal productivity in systems

that adopt deferred grass pasture as the basic diet. This study aimed to evaluate the effect
of levels of supplementation of mesquite pod meal on intake, digestibility of nutrients,
and body weight (BW) gain of goats in grazing deferred. Thirty-five (35) goat males (24.0
± 2.9 kg BW) were distributed in a completely randomized design, with five treatments
and seven replications. Treatments constituted a supplement control (0.05% BW of

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 Rev Mex Cienc Pecu 2023;14(3):523-538

protein-mineral salt) and increasing levels at 0.5; 1.0; 1.5, and 2.0% BW of
supplementation with mesquite pod meal. Dry matter intake and nutrients of the forage
and total diet increased linearly (P<0.0001) as a function of the levels of the supplement.
A quadratic effect was observed (P<0.0001) for the digestibility variables, except for
ethereal extract and non-fiber carbohydrates, which presented an increasing linear effect
(P<0.0001). Supplementation levels increased linearly on the performance variables
(P<0.05). It is recommended the concentrate supplementation at 2.0% BW with mesquite
pod meal at 30 % in its composition for promoting the reduction of the productive cycle
of goats in deferred Urochloa grass pastures.

Keywords: Alternative supplementation, Animal production, Pasture management,

Prosopis juliflora, Semiarid, Weight gain.

Received: 16/12/2021

Accepted: 21/02/2023

Introduction

Goats breeding for meat production in the northeast region of Brazil is predominantly
extensive and seasonal because the rainfall distribution is irregular and the adverse
edaphoclimatic characteristics affect the forage production(1). The quantity and the quality
of the food become obstacles to the productive chain, especially during the dry season. In
this critical period, animals lose body weight, thus delaying the age at slaughter, causing
losses to producers and to the economy in general.

These conditions justify the use of pasture management techniques such as deferral,
which consists in selecting a pasture area of the property and excluding grazing, usually
at the end of the rainy season(2), so that forage accumulation occurs to be grazed during
the dry period, minimizing the negative effects of seasonal forage production on animal
productivity(3). Nonetheless, changes in pasture structure occur during the deferment
period and are also characterized by low nutritional value as a consequence of changes in
environmental conditions and of the forage plant phenology itself, which tend to reduce
the performance of ruminants(4).

Supplementation emerges as a simple strategy used for both an attempt to address the
nutritional deficiencies of the pasture, by providing the balance of the animal’s diet, and
also to reduce the risk caused by fluctuating pasture dry matter production(5,6). However,
the effect of adding highly degradable carbohydrates to forage-based diets can be
beneficial or undesirable, depending on the source used and, above all, the amount eaten

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by the animals(7). The constant search for alternative feeds to corn, which is the most used
energy concentrate in animal production systems, is fundamental, especially in regions
distant from those producing grain.

The use of mesquite pod meal (Prosopis juliflora (Sw.) D.C.) as a substitute for corn
becomes an alternative due to its easy accessibility in semiarid regions and its use in diets
for small ruminants has shown better productive indices compared to corn(8-13). However,
the consumption of P. juliflora pods as the main source of food causes intoxication in
animals(14-16). In this context, the objective of this study was to evaluate the effect levels
of concentrate with mesquite pod meal on the performance of goats in deferred Urochloa
grass pastures.

Material and methods

Ethical principles of experimentation

All the animal care and handling procedures were approved by the Ethics Committee on
Animal Use f the State University of Southwest Bahia – UESB, with protocol number
23/2017.

Experimental area

The experiment was conducted at the Research Center for Sheep and Goat, located in the
Iaçu municipality, State of Bahia, Brazil. The experimental period was from April 2018
to July 2018 with mean rainfall at 31.2 mm. The climate of the region is characterized as
a tropical climate with a dry season(17).

Experimental procedures, animals, and diets

The total pasture area was 4.4 ha composed exclusively of Urochloa grass (Urochloa
mosambicensis (Hack) Daudy). The pasture was fenced for 110 d and used from April of
the same year until July 2018 (92 d). The employed grazing method was continuous
stocking with a variable stocking rate. The evaluated treatments were five supplements:
protein-mineral salt and levels of concentrate containing mesquite pod meal. The
structural variables of the deferred pasture were evaluated every 23 d during the whole
usage period. Pasture height was measured with a graduated ruler in centimeters, with

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100 readings performed per picket at the average curvature height of the leaves. Forage
mass was estimated by cutting close to the forage soil (12 samples) with a square of 0.25
m2 as described by McMeniman(18) and the pasture composition was measured (Table 1).
Thirty-five uncastrated male goats of the Boer breed, at approximately 4-mo old, an initial
body weight (BW) of 24.0 ± 2.9 kg were used, distributed in a completely randomized
design with five treatments and, 7 replicates were adopted. The animals were kept in a
Urochloa grass pasture under continuous stocking during the day (0007 to 1600 h) and
housed in sheds in collective stalls during the night where they received: protein-mineral
salt at fixed at 0.05% BW (control) and increasing levels at 0.5, 1.0, 1.5, and 2.0% BW
of mesquite pod meal as an energy supplement.

Table 1: Pasture composition

Green leaf, g kg-1 240.38

Green stem, g kg-1 530.00
Senescent material, g kg-1 670.44
Leaf/stem ratio 0.460
Availability of dry matter (DM), kg ha-1 3.264
kg DM leaf ha-1 795.76
kg DM stem ha-1 1.729.92
kg DM senescent material ha-1 737.99

The supplements were formulated to meet the protein requirements for maintenance and
to provide an average daily gain of 150 g, according to the NRC(19). Table 2 shows the
chemical composition of the supplements and Urochloa grass. A 15-d adaptation period
was used for the animals to acclimatize to the supplement and to the research facilities,
followed by 92 d of the experiment divided into four subperiods of sample collection that
lasted 5 d.

Table 2: Composition (g 100 g-1 of DM) of the supplements in ingredients and nutritive
value of protein-mineral salt, Urochloa grass and concentrate containing mesquite pod
meal
Ingredient Supplement
Concentrate Protein-mineral
Corn meal 45.0 33.6
salt
Soybean meal 22.0 20.0
Mesquite pod meal 30.0 -
Urea 2.0 9.1
Mineral salta 1.0 13.6
Ammonium sulfate - 1.0
Sodium chloride - 22.7
Total 100.0 100.0

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Nutrient Protein-mineral salt Urochloa grass Concentrate

Dry matter 88.6 91.3 86.1

Organic matter 72.2 89.1 98.8
Crude protein 41.2 13.7 20.3
Ether extract 1.5 1.9 2.6
Total carbohydrates 29.4 73.5 75.9
Non-fiber 44.3 13.1 45.4
NDF free of ash protein
carbohydrates 13.2 60.4 36.2
Acid detergent fiber 7.6 43.0 28.2
Indigestible NDF 2.9 12.4 4.8
Lignin 1.4 7.6 13.7
Ash 27.8 10.9 1.2
a
Quantity/kg of product: Calcium (max.)= 120 g; phosphorus= 87 g; copper= 590 mg; cobalto= 40 mg;
iodine= 80 mg; manganese= 1,300 mg; molybdenum= 300 mg; fluorine (max)= 870 mg.
NDF= neutral detergente fiber.

Mature pods were obtained after harvesting in the ground, manually selected, discarding
those attacked by insects, fungi, and of a small development. The pods were dried sun
drying was used. And then, processed in a Wiley knife mill (A. H. Thomas, Philadelphia,
PA, USA) using a 1-mm sieve, to obtain the pod meal.

Evaluation of intake, digestibility, and live weight gain

The dry matter (DM) intake of forage and digestibility of nutrients was estimated from
the fecal output, with the use of Enriched and Purified Isolated Lignin from Eucalyptus
Grandis (LIPE®; Belo Horizonte, MG, Brazil) as an external marker(20), and indigestible
acid detergent fiber (iADF) as an internal marker. The DM intake per supplement was
estimated using titanium dioxide. The titanium dioxide (TiO2) was analyzed according to
the methodology described by Titgemeyer(21). Titanium dioxide was mixed with the
supplement and supplied in the amount of 5 g per animal. LIPE® capsule oral
administration for each animal happened for 7 consecutive days; the first 2 d were to
stabilize the fecal excretion of the marker(20,22). Fecal samples were collected directly
from the rectum twice a day (0800 and 1700 h), for 5 d, and stored in a cold chamber at -
10 °C.

The concentration of iADF in supplement samples, consumed forage, and feces were
obtained after incubation in situ for 264 h according to Casali et al(23). The voluntary
intake of DM was estimated by the ratio between fecal excretion and indigestibility from
the internal indicator iADF, as described above, using the equation proposed by
Detmann(24):

DMI = {[(FE × MCF) – CIS] /CIFOR} + DMIS

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Where: DMI= dry matter intake (kg d-1); FE= fecal excretion (kg d-1); MCF= marker
concentration in the animal feces (kg kg-1); CIS= concentration of iADF in the
supplement (kg d-1); CIFOR = concentration of iADF in forage (kg kg-1); and DMIS=
intake of supplement DM (kg d-1).

Supplement intake was measured by the quantity supplied divided by the number of
animals in the treatment. The estimate of the quality of forage consumed was performed
by analyzing the samples, using the technique for manual simulation of grazing(25), by
visual observation of the animals.

The animals were weighed at the start, every 23 d, and at the end of the experiment. At
the beginning of the experimental period, the animals were subjected to a 16-h solid fast
and weighed to determine initial body weight (IBW). Total weight gain (TWG) was
estimated as the difference between final body weight (FBW) and initial body weight
(IBW): TWG= (FBW - IBW). Average daily gain (ADG) was calculated by dividing
TWG by the total number of days in the experiment: ADG= TWG/days in the experiment.
Finally, the feed conversion ratio was calculated as the ratio between dry matter intake
(kg d-1) and TWG (kg d-1).

Sample processing and laboratory analyses

The contents of DM (method INCT-CA G - 003/ 1), ash (method INCT-CA M-001/1),
crude protein (CP) (method INCT-CA N-001/1), ether extract (EE) (method INCT-CA
G-004/1) were determined in the forage and supplement samples, according to the
recommendations described by AOAC(26). For the neutral detergent fiber (NDF) analyses,
the samples were treated with thermostable alpha-amylase, without the use of sodium
sulfite, and corrected for residual ash(27). The correction of NDF for the nitrogen
compounds and the estimate of the concentration of nitrogen-neutral detergent insoluble
compounds (NDIN) and acid (ADIN) were performed according to Licitra(28).

Total carbohydrates (TC) were estimated according to Sniffen(29), non-fibrous

carbohydrates were calculated according to the methodology proposed by Hall(30), using
NDFap and total digestible nutrients (TDN) were calculated according to Weiss(31), but
using NDF and NFC corrected for ash and proteins.

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Statistical analysis

The statistical analysis of the data was achieved by the MIXED procedure of the SAS
statistical computer program (SAS, 2006), considering a mixed model. The data were
submitted for analysis of variance (ANOVA) and was realized the contrast between the
control treatment with supplementation levels of concentrate. Also, the polynomial
contrast and regression analysis were performed for supplementation levels (0.5, 1.0, 1.5,
and 2.0% BW), adopting a 5 % to 10 % probability for type 1 error.

Results

The DM intake and of the nutrients of the total diet, forage (deferred Urochloa grass), and
supplements were greater (P<0.0001) for the animals who received the supplementation
mesquite pod meal, independently of the levels, compared to animals fed only with
protein-mineral salt, due to the higher supply of nutrients from concentrate (Tables 3, 4,
and 5). The nutrient concentrations were proportionally unchanged as a function of DM
intake, independent of supplementation levels since the concentrate supplement was the
same.

Table 3: Nutrient intakes of the diet of goats on grazing of deferred Urochloa grass
with supplementation levels
Supplementation P value
Concentrate level (% BW)
SE Contrast L Q
Item PMS 0.5 1.0 1.5 2.0
Total intake (g d-1)
DM 353. 506. 936. 1196. 1738. 93. <0.0001 <0.000 0.590
CP 52.9
008 80.0
3 151.
4 196.7
6 286.7
2 15.
6 <0.0001 <0.000
1a 0.614
7
EE 6.6 10.6 20.1 26.0 37.8 2.1 <0.0001 b
<0.000 0.608
6 8 1 0
NFC 51.2 113. 235. 316.0 464.2 27. <0.0001 c
<0.000 0.663
1 9
NDFa 206. 270. 480. 603.2 872.2 45. <0.0001 d
<0.000 0.563
3 8 4 1 1
TND 80.0 260. 640. 890.0 1390. 80. <0.0001 e
<0.000 0.516
p 2 5 6 3 1 3
Total intake (g kg0-1 BW) 0 0 0 1 f
8
DM 13.8 17.4 31.7 40.1 54.0 2.9 <0.0001 <0.000 0.958
NFCa 8.0 9.3 16.3 20.2 27.1 1.4 <0.0001 1g
<0.000 0.983
3
-1 0.75 h
Total
p intake (g kg BW ) 1 6
CP 4.6 6.3 12.0 15.4 21.2 1.2 <0.0001 <0.000 0.954
NFC 4.5 9.0 18.6 24.7 34.3 2.0 <0.0001 1i
<0.000 0.997
9
j
PMS= protein-mineral salt; SE= mean standard error; Contrast= PMS vs supplementation levels;
1 6 L=
linear effect; Q= quadratic effect; BW= body weight; DM= dry matter; CP= crude protein; EE= ether
extract; NFC= non-fiber carbohydrates; NDFap= neutral detergent fiber corrected for ash and protein;
TDN= total digestible nutrients; Significant * (P<0.0001); ** (P<0.001); *** (P<0.01); **** (P<0.05); ns
(P>0.05); aŶ= 110.68ns + 792.82 X *; bŶ= 12.4414ns + 133.69X *; cŶ= 1.8056 ns + 17.5525 X *; dŶ=
(0.5534ns + 226.31X *; eŶ= 77.0549 ns + 387.13X *; fŶ= - 0.099 *** + 0.725X *; gŶ= 5.7387ns + 24.3310 X
; Ŷ= 3.6930 **** + 11.7801 X *; hŶ= 12.1673ns + 58.1458X *; iŶ= 15091ns + 9.8537X *; jŶ= 0.7727ns +
* g

16.8385X *

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Table 4: Nutrient intake of forage of goats on grazing of deferred Urochloa grass with
supplementation levels
Supplementation P value
Concentrate level (% BW)
Item SE Contrast L Q
PMS 0.5 1.0 1.5 2.0
-1
Forage intake (g d )
DM 338.2 361.0 586.3 704.0 1006.5 47.4 <0.0001 <0.0001a 0.5011
CP 46.5 49.7 80.6 96.8 138.4 6.5 <0.0001 <0.0001b 0.5011
EE 6.4 6.8 11.1 13.3 19.0 0.9 <0.0001 <0.0001c 0.5011
NFC 44.3 47.3 76.9 92.3 131.9 6.2 <0.0001 <0.0001d 0.5011
NDFap 204.2 218.0 354.0 425.0 607.7 28.6 <0.0001 <0.0001e 0.5011
Forage intake (g kg-1 BW)
DM 13.2 12.5 19.9 23.6 31.3 1.5 <0.0001 <0.0001f 0.9504
NDFap 7.9 7.5 12.0 14.3 18.9 0.9 <0.0001 <0.0001g 0.9504
Forage intake (g kg-1 BW0.75)
CP 4.1 4.0 6.4 7.6 10.3 0.5 <0.0001 <0.0001h 0.8407
NFC 3.9 3.8 6.1 7.2 9.8 0.5 <0.0001 <0.0001i 0.8407
PMS= protein-mineral salt; SEM= mean standard error; Contrast= PMS vs supplementation levels; L=
linear effect, Q= quadratic effect, BW= body weight; DM= dry matter; CP= crude protein; EE= ether
extract; NFC= non-fiber carbohydrates; NDFap= neutral detergent fiber corrected for ash and protein;
TDN= total digestible nutrients; Significant * (P <0. 0001); ** (P <0.001); *** (P <0.01); **** (P <0.05); ns
(P >0.05); aŶ= 152.70 *** + 414.60X *; bŶ = 21.0037*** + 57.0255X *; cŶ = 2.8887 *** + 7.8429X ****; d=
20.0181 *** + 54.3495X *; eŶ = 92.1912 *** + 250.30 X *; fŶ = 6.6600*** + 12.3386X *; gŶ = 4.0208*** +
7.4491 X *; hŶ = 2.0117*** + 4.0883 X *; iŶ = 1.9173 *** + 3.8965X *

Table 5: Nutrient intake of concentrate of goats on grazing deferred Urochloa grass

pasture with supplementation levels

Supplementationn P value
Item PMS Concentrate level (% BW) SE Contrast L Q
0.5 1.0 1.5 2.0
Concentrate intake (g d-1)
DM 15.6 145.2 350.1 492.6 731.6 47.2 <0.0001 <0.0001a 0.7327
CP 6.4 29.4 71.0 99.8 148.3 9.4 <0.0001 <0.0001b 0.7327
EE 0.2 3.7 8.98 12.6 18.8 1.2 <0.0001 <0.0001c 0.7327
NFC 6.9 66.0 159.0 223.7 332.2 21.4 <0.0001 <0.0001d 0.7327
NDFap 2.1 52.5 126.6 178.1 0.73 17.2 <0.0001 <0.0001e 0.7327
Concentrate intake (g kg-1 BW)
DM 0.6 5.0 11.8 16.5 22.7 1.5 <0.0001 <0.0001f 0.8462
NDFap 0.07 1.8 4.3 6.0 8.2 0.6 <0.0001 <0.0001g 0.8462
Concentrate intake (g kg BW0. 75)
-1

CP 0.6 2.3 5.6 7.8 10.9 0.7 <0.0001 <0.0001h 0.9460

NFC 0.6 5.2 12.5 17.5 24.5 1.6 <0.0001 <0.0001i 0.9460
PMS= protein-mineral salt; SE= mean standard error; Contrast= contrasts between the SP and the levels of
supplementation; L= linear effect, Q= quadratic effect; BW= body weight; DM= dry matter; CP= crude protein; EE=
ether extract; NFC= non-fiber carbohydrates; NDFap= neutral detergent fiber corrected for ash and protein; *P<0.
0001; ** P<0.001; *** P<0.01; **** P<0.05; ns P>0.05; aŶ= - 43.8774 ns + 379.65 X *; bŶ = - 8.8931ns + 76.9487X *; cŶ
= - 1.1263ns + 9.7457 X *; dŶ = - 19.9255 ns + 172.41X*; eŶ = - 15.8682 ns + 137.30X *; fŶ = -0.9681 ns +12.0713X*;
gŶ = - 0.3501 ns + 4.3656X*; hŶ = - 0.5191 * + 5.7915 X; iŶ = - 1.1631ns + 12.9761 X *

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The dry matter digestibility and of the other nutrients were greater (P<0.0001) for
supplementation levels compared to protein-mineral salt. There was a quadratic effect for
the digestibility of most nutrients, except for EE and NFC which showed a linear increase
(Table 6). Maximum points were calculated for digestibilities of DM, OM, CP, and NDF
near the upper limit of supplementation (2.0% BW), with the same response for the
variation of TDN content. Therefore, it was not possible to estimate the maximum point
because the range of the supplementation levels studied was restricted to a range of the
quadratic fit in which the rate of increase in digestibility coefficient was not proportional
to the supplement increment, that is, it was in points previous to curve inflection.

Table 6: Nutrient digestibility (g 100 g-1 of DM) of goats on grazing deferred Urochloa
grass pasture with supplementation levels
Supplementation P value
Concentrate level (%
SE Contrast L Q
PMS BW)
Item
0.5 1.0 1.5 2.0
DM 21.8 45.6 66.2 73.2 80.0 3.8 <0.0001 <0.0001 0.0011a
OM PMS 48.1
21.9 68.3 74.7 81.0 3.8 <0.0001 <0.0001 0.0008b
NDFap 23.7 43.2 63.0 69.7 77.5 3.5 <0.0001 <0.0001 0.0081c
EE 40.4 43.9 60.3 68.1 76.4 2.7 <0.0001 <0.0001d 0.2183
CP 31.1 48.8 68.1 75.8 79.4 3.3 <0.0001 <0.0001 0.0002e
NFC 10.7 57.5 82.4 86.3 90.4 5.3 <0.0001 <0.0001f 0.0056
TDN 21.6 47.1 67.0 73.6 79.9 3.8 <0.0001 <0.0001 0.0008g
PMS= protein-mineral salt; SEM= mean standard error; Contrast= PMS vs supplementation levels; L=
linear effect; Q= quadratic effect; BW= body weight; DM= dry matter; CP= crude protein; EE= ether
extract; NFC= non-fiber carbohydrates; NDFap= neutral detergent fiber corrected for ash and protein;
TDN= total digestible nutrients; Significant **(P<0. 0001); ** (P<0.001); *** (P<0.01); ****(P<0.05); ns
(P>0.05); aŶ= 20.099* + 58.357 X * - 14.294 X2 **; bŶ= 23.499* + 56.554 X * - 13.993 X2 **; cŶ=19.817 **
+ 52.997 X * - 12.166 X2 ***; dŶ= 39.7744* +/ 18.596 X *; eŶ= 23.452 * + 58.868 X * - 15.490 X2 **; fŶ=
72.547* + 8.996 X *; gŶ= 22.343* + 56.718 X* - 14.070 X2 **

The supplementation levels with the energy source containing mesquite pod meal
provided greater final body weight (FBW), average daily gain (ADG), and total weight
gain (TWG) when compared to the protein-mineral salt supplementation (Table 7). There
was a linear effect for supplementation levels on the performance variables (P<0.05).
Supplementation levels promoted a linear increase (P<0.0001) in feed conversion and the
level at 0.5% BW was efficient considering 92 d of pasture supplementation to reach 35
kg for slaughter weight.

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Table 7: Performance of goats on grazing deferred Urochloa grass pasture with

supplementation levels for 92 days
Supplementation P value
Concentrate level (% BW)
SE Contrast L Q
Item PMS
0.5 1.0 1.5 2.0
IBW 23.3 24.1 24.2 23.7 24.4 0.5 P value
0.4669 0.9372 0.7711
FBW 28.9 34.6 34.6 37.6 40.3 0.8 <0.0001 0.0004a 0.3039
BW 26.1
PMS 29.4 29.4 30.7 32.4 0.6 0.0001 0.02802b 0.4471
0.75
BW 11.5 12.6 12.6 13.0 13.6 0.2 0.0002 0.02810c 0.4437
TWG 5.7 10.5 10.4 13.9 15.9 0.7 <0.0001 0.0003d 0.3311
ADG 0.06 0.12 0.12 0.16 0.18 0.01 0.0001 0.0003e 0.3312
FC 22.7 4.4 8.1 8.0 10.0 3.4 0.3877 < 0.1987
f levels; L=
PMS= protein-mineral salt; SE= mean standard error; Contrast= PMS vs supplementation
0.0001
linear effect; Q= quadratic effect; BW= body weight; IBW= initial body weight (kg); FBW= final body
weight (kg); TWG= total weight gain (kg); ADG= average daily gain; FC= feed conversion (kg DMI/ kg
BW); Significant *(P<0.0001); ** (P<0.001); ***(P<0.01); ****(P< .05); ns (P>0.05); aŶ= 31.560 * + 4.263X
; Ŷ= 27.812* + 2.132 X ****; cŶ= 12.112 * + 0.681X ****; dŶ= 3.016 * + 4.273X *; eŶ= 0.574 ns + 0.047 X
* b

; Ŷ= 0.666 *** + 1.234 X*

* f

Discussion

The concentrate supplement provided an improvement of nutrient supply to the ruminal

microbiome, leading to greater digestion of the fiber, which consequently promoted an
increase in total and forage DM intake characterizing the additive effect (Tables 3, 4, and
6). Moore(32) stated that if supplement intake does not influence forage intake, the
substitution coefficient is zero and, when positive, it means that forage intake was
increased by supplementation. This fact can be explained by the relationship between the
total digestible nutrients and the crude protein (TDN/CP) of the forage, which were 0.4,
1.8, 3.3, 4.0, and 4.7 for respective supplementation levels.

However, the increase in DM intake of forage supplemented with concentrate 0.5% BW

was 6 % higher than protein-mineral salt supplementation. Still, when corrected for BW,
the DM intake of forage was 5 % lower. It indicates that the supplementation at 0.5% BW
was insufficient to avoid the ingestion's empty physical effect. Due to the increase in DM
intake, the nutrient intake of CP, NDFap, and NFC also increased (Table 4).

For concentrate supplementation levels, there was an increase in forage intake of 12.3 g
kg-1 BW for each concentrate percentual unit. As the supply of concentrate was
controlled, it can be evidenced that the 0.5% BW supplied would not be indicated to
increase the forage intake, despite the improvement of fiber and other nutritional
components' digestibilities (Table 6). The forage proportions were 95.59, 71.31, 62.61,
58.83, and 57.91 % in the respective diets with protein-mineral salt and concentrate levels.

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In the deferred pasture, there is usually a decrease in CP and fiber digestibilities, because
of the maturation process. In this study, the CP content of the Urochloa grass was 13.7 %
and 85.8 % was in the NDF fraction, presenting a lower rumen degradation rate,
especially when the microbial growth was affected by the lower content of soluble
nutrients. The forage contributed to decreasing proportion of CP according to the
supplementation levels, whose respective values were 13.1; 9.8; 8.6; 8.1, and 7.9 %.
Likewise, the proportional values of NDF from forage also decreased: 57.7; 43.1; 37.8;
35.5, and 35.0 %, respectively.

The concentrate supplementation levels provided an increase in CP, NFC, and TDN
intakes, being that the CP concentration in the total DM ingested was similar when
comparing the supplementation with protein-mineral salt and levels of concentrate (Table
3). In addition, it was observed that, regardless of the use of protein-mineral salt or
concentrate levels, goat’s kids did not change the composition of the forage consumed,
with an average of 13.7 % CP, 13.1 % NFC, and 60.4 % NDFap, characterizing non-
selectivity during grazing (Table 4).

The greater supply of concentrate containing mesquite pod meal at 30 %, improved the
rumen fermentation and digestibility of nutrients. The maximum points for digestibility
were estimated for DM, OM, NDF, and CP in the range of 1.9 % to more than 2.0% BW
in supplementation, with the same changing, for the TDN concentration. The quadratic
fit was possible because the digestibility did not change proportionally to the increase in
supplementation, probably due to the increase in the rumen passage rate. The linear
increase in the digestibility of EE and NFC is consistent with the fact that there was an
increase in the intake of these nutritional components, due to the levels of
supplementation and, thus, the increment of intestinal utilization.

It can be inferred that the rumen passage rate affected the CP and NDF digestibilities
because the highest proportion of CP from forage belongs to the fibrous fraction. The
greater passage rate can reduce the extent of ruminal degradation of the fibrous fraction
of the diet when the DMI rises. Considering that this fraction of the diet is not effectively
digested in the small intestine.

To increase the intake of forage, it is necessary to manipulate the diet through two
mechanisms, increasing the ruminal digestion rate and/or accelerating the rate of passage
of indigestible components(33,34). In this study, it was observed that the supplementation
with concentrate provided an increase in forage intake, as a consequence of both increased
ruminal digestion rates, since the NDF digestibility increased. The supplementation with
concentrate has an associative effect with the pasture, that is, it entails changes in
digestibility (Table 6) and/or in forage intake (Table 3), which may have additive and
substitutive effects. An additive effect was observed because there was an increase in
TDN intake as a consequence of a greater intake of concentrate, without a decrease in
forage intake (Table 3).

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The major intake of mesquite pod meal occurred at 2% supplementation showing an

average of 219.5 g d-1, which is equivalent to 126.2 g kg-1 of total DM consumed. Studies
have reported that the use of mesquite pod meals in diets should not exceed 200 g kg-1 of
DM consumed in goats, both for BW gain and for better lactating performance. Therefore,
in this study, the toxic effect of mesquite pod meal did not occur, since the level of 2.0%
BW of the concentrate supplement showed a greater ADG (Table 7).

There was a linear effect of supplementation levels on the performance variables (P<0.05)
because the concentrate supplementation levels increased the total DM intake and
improved the digestibility, reflecting a greater ADG.

The supplementation with protein-mineral salt resulted in reduced ADG as a response to

the restriction of its intake and the advanced stage of maturation of the Urochloa grass,
which presented high contents of NDF indigestible and CP bound to the fibrous fraction
(Table 1). However, animals kept in grazing under semiarid conditions usually present a
loss of BW during the critical period of forage production. Thus, the use of simple
technology, such as supplementation with protein-mineral salt, softens the effects of low
availability and quality of biomass. Additionally, the supplementation levels with
concentrate provided higher ADG when compared to protein-mineral salt
supplementation, and the levels of 1.5% and 2.0% BW provided 155 and 176 g in the
ADG, respectively.
The DM intake has an influence on performance, as it determines the number of nutrients
ingested, which are necessary to meet the requirements of maintenance and animal
production. The feed conversion at 0.5% BW of mesquite pod meal supplementation did
indicate the best productive response than the other levels since the goat kids reached the
ideal slaughter weight (35 kg) at 92 d of supplementation.

Conclusions and implications

The use of deferred pasture enables a high forage supply in the dry season of the year,
even if it is of low quality; and when combined with minimum supplementation levels
(0.5% BW), it is possible to increase the rate of average daily gain contributing to the
reduction of the productive cycle. The mesquite pod meal supplementation at 2.0% BW
provides a higher average daily gain and slaughter body weight at 92 d under grazing,
enabling greater gain per pasture area.

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Acknowledgements

The authors thank the State University of Southwest Bahia for providing the physical
infrastructure and staff necessary for this study. We are also grateful and financial support
and grants from the Brazilian National Research Council (CNPq).

Literature cited:
1. Gurgel ALC, Difante GS, Emerenciano Neto JV, Roberto FFS, Zaros LG, Costa MG,
Ítavo LCV, Ítavo CCBF. Impact of supplementation with different protein sources
on the parasitological profile of ovine matrices and development of lambs. Biosci J
2020;36(2):496-506.

2. Shio AR, Veloso CM, Silva FF, Ìtavo LCV, Mateus RG, Silva RR. Ofertas de
forragem para novilhas nelore suplementadas no período de seca e transição
seca/águas. Acta Sci Anim Sci 2011;33(1):9-17.
https://doi.org/10.4025/actascianimsci.v33i1.9112.

3. Santos MER, Castro MRSA, Gouvêia SC, Gomes VM, Fonseca DM, Santana SS.
Contribuição de perfilhos aéreos e basais na dinâmica de produção de forragem do
capim-braquiária após o pastejo diferido. Biosci J 2014;30(1):24-430.

4. Euclides VPB, Montagner DB, Macedo MCM, Araújo AR, Difante GS, Barbosa RA.
Grazing intensity affects forage accumulation and persistence of Marandu
palisadegrass in the Brazilian savannah. Grass Forage Sci 2019;74:450-462.
https://doi.org/10.1111/gfs.12422.

5. Fernandes LS, Difante S, Montagner DB, Emerenciano Neto JV, Araújo IMM,
Campos NRF. Structure of massai grass pasture grazed on by sheep supplemented
in the dry season. Grassl Sci 2017;1-7. https://doi.org/10.1111/grs.12165.

6. Geron LJV, Mexia AA, Garcia J, Silva MM, Zeoula LM. Suplementação
concentrada para cordeiros terminados a pasto sobre custo de produção no período
da seca. Semina: Ciênc Agrár 2012;33(2):797-808. https://doi.org/10.5433/1679-
0359.2012v33n2p797.

7. Nascimento ML, Paulino MF, Detmann E, Leão MI, Valadares Filho SC, Henriques
LT. Fontes de energia em suplementos múltiplos para novilhos em pastejo durante o
período das águas. Rev Bras Zootec 2010;39(4):861-872.

8. Santos EJ, Pereira MLA, Cruz JF, Figueiredo MP, Almeida PJP, Novaes EJ, Souza
ACS, Alencar DO, Sousa LB, Pereira TCJ. Crude protein levels in diets containing
pelleted concentrate for lactating goats: intake, digestibility, milk production and
composition. Semina: Ciênc Agrár 2015;36(4):2849-
2860.https://doi.org/10.5433/1679-0359.2015v36n4p2849.

535
 Rev Mex Cienc Pecu 2023;14(3):523-538

9. Santos EJ, Pereira MLA, Almeida PJP, Pereira TCJ, Chagas DMT, Silva TVBS.
Excreções de derivados de purina obtidos por duas metodologias de coleta de urina
em ovinos alimentados com farelo da vagem de algaroba em substituição a silagem
de capim Elefante. Rev Eletr Nutritime 2015; 12.

10. Pereira TCJ, Pereira MLA, Almeida PJP, Carvalho GGP, Silva FF, Silva HGO,
Santos AB. Substitution of corn for mesquite pod meal in diets for lambs. Ital J Anim
Sci 2014;13:3278. https://doi.org/10.4081/ijas.2014.3278.

11. Santos AB, Pereira MLA, Silva HGO, Pedreira MS, Carvalho GGP, Ribeiro LSO,
Almeida PJP, Pereira TCJ, Moreira JV. Nitrogem metabolism in lactating goats fed
with diets containing different protein sources. Asian Australas J Anim Sci
2014;27(5):658-666. https://doi.org/10.5713 / ajas.2013.13493.

12. Pereira TCJ, Pereira MLA, Oliveira CAS, Argôlo LS, Silva HGO, Pedreira MS,
Almeida PJP, Santos AB. Mesquite pod meal in diets for lactating goats. Rev Bras
Zootec 2013:42(2):102-108. https://doi.org/10.1590/S1516-35982013000200004.

13. Almeida PJP, Pereira MLA, Silva FF, Santos AB, Pereira TCJ, Santos EJS, Moreira
JV. Santa Inês sheep supplementation on Urochloa grass pasture during the dry
season: intake, nutrient digestibility and performance. Rev Bras Zootec
2012;41(3):668-674. http://dx.doi.org/10.1590/S1516-35982012000300029.

14. Silva VDA, Silva AMM, Silva JHC, Costa SL. Neurotoxicity of Prosopis juliflora:
from natural poisoning to mechanism of action of its piperidine alkaloids. Neurotox
Res 2018. https://doi.org/10.1007/s12640-017-9862-2.

15. William K, Jafri L. Mesquite (Prosopis juliflora): livestock grazin, its toxicity and
management. J Bior Management 2015;2(2):49–58.
https://doi.org/10.35691/JBM.5102.0021.

16. Tabosa IM, Riet-Correa F, Barros SS, Summers BA, Simões SVD, Medeiros RMT,
Nobre VMT. Neurohistologic and ultrastructural lesions in cattle experimentally
intoxicated with the plant Prosopis juliflora. Vet Pathol 2006;43(5):695–701.
https://doi.org/10.1354/vp.43-5-695.

17. Thorthwaite CW. An approach toward a rational classification of climate. Geogr Rev
1948;38:55-94.

18. McMeniman NP. Methods of estimating intake of grazing animals. In: Reunião
Anual da Sociedade Brasileira de Zootecnia. Juiz de Fora. Anais Juiz de Fora:
Sociedade Brasileira de Zootecnia. 1997;34:131-168.

19. National Research Council. Nutrient requirements of small ruminants: Sheep, goats,
cervids and New World camelids. National Academies Press, Washington, DC.
2007.

536
 Rev Mex Cienc Pecu 2023;14(3):523-538

20. Saliba EOS, Faria EP, Rodriguez NM, Moreira GR, Sampaio IBM, Saliba JS,
Gonçalves LC, Borges I, Borges ALCC. Use of infrared spectroscopy to estimate
fecal output with Marker LIPE®. Int J Food Sci Nutri 2015;S4:001:1-10.
https://dx.doi.org/10.19070/2326-3350-SI04001.

21. Titgemeyer EC, Armendariz CK, Bindel DJ, Greenwood RHCA, Loest CA.
Evaluation of titanium dioxide as a digestibility marker for cattle. Animal Sci J
2001;79(4):1059-1063. https://doi.org/10.2527/2001.7941059x.

22. Freitas TB, Felix T, Pedreira MS, Silva RR, Silva FF, Silva HGO, Tigre JS.
Replacement of soybean meal with treated castor bean meal in supplements for
grazing lambs. Rev Bras Saúde Prod Anim 2017;18(3):465–478.
https://doi.10.1590/s1519-99402017000300007.

23. Casali AO, Detmann E, Valadares Filho SC, Pereira JC, Henriques LT, Freitas SG,
Paulino MF. Influência do tempo de incubação e do tamanho de partículas sobre os
teores de compostos indisgestíveis em alimentos e fezes bovinas obtidos por
procedimento in situ. Rev Bras Zootec 2008;37(2):335-342.
http://dx.doi.org/10.1590/S1516-35982008000200021.

24. Detmann E, Paulino MF, Zervoudakis JT, Valadares Filho SC, Euclydes RF, Lana
RP, Queiroz SQ. Cromo e indicadores internos na estimação do consumo de novilhos
mestiços suplementados a pasto. Rev Bras Zootec 2001;30(5):1600-1609.
https://doi.org/10.1590/S1516-35982001000600030.

25. Euclides VPB, Macedo MCM, Oliveira MP. Avaliação de diferentes métodos de
amostragem sob pastejo. Ver Bras Zootec 1992;21(4):691-702.

26. AOAC. Official methods of analysis. 16th ed. Association of Official Analysis
Chemists, Arlington, Virginia. 1995.

27. Mertens DR. Gravimetric determination of amylase-treated neutral detergent fiber in

feeds with refluxing in beakers or crucibles: collaborative study. J AOAC Int
2002;85(6):1217-1240.

28. Licitra G, Hernandez TM, Van Soest PJ. Standardization of procedures for nitrogen
fracionation of ruminant feed. Anim Feed Sci Technol 1996;57(4):347-358.
https://doi.org/10.1016/0377-8401(95)00837-3.

29. Sniffen CJ, O´Connor JD, Van Soest PJ, Fox DG, Russell JB. A net carbohydrate
and protein system for evaluating cattle diets: II. Carbohydrate and protein
availability. J Anim Sci 1992;70(11):3562-3577. https://doi.org/10.2527 /
1992.70113562x.

30. Hall MB. Neutral detergent-soluble carbohydrates. Nutritional relevance and

analysis. Gainesville: University of Florida. 2003.

537
 Rev Mex Cienc Pecu 2023;14(3):523-538

31. Weiss W. Energy prediction equations for ruminant. 1999. In: Cornell Nutrition
Conference for Feed Manufacturers, 61. Ithaca. Proc Cornell University. Ithaca.
1999:176-185.

32. Moore JE, Brant MH, Kunkle WE, Hopkins DI. Effects of supplementation on
voluntary forage intake, diet digestibility, and animal performance. J Anim Sci
1999;77(Suppl 2):122-135.

33. Costa VAC, Detmann E, Valadares Filho SC, Paulino MF, Henriques LT, Mantovani
HC. Degradação in vitro da fibra em detergente neutro de forragem tropical de baixa
qualidade em função de suplementação com proteína e/ou carboidratos. Rev Bras
Zootec 2008;37(3):494-503. https://doi.org/10.1590/S1516-35982008000300015.

34. Costa VAC, Detmann E, Valadares Filho SC, Paulino MF, Henriques LT, Mantovani
HC. Degradação in vitro da fibra em detergente neutro de forragem tropical de alta
qualidade em função de suplementação com proteína e/ou carboidratos. Rev Bras
Zootec 2009;38(9):1803-1811. https://doi.org/10.1590/S1516-35982009000900024.

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 https://doi.org/10.22319/rmcp.v14i3.6251

Article

Genetic parameters, correlations and trends of reproductive traits in

Holstein cattle from Mexico

Carolina Durán-Alvarez a,b,c

Adriana García-Ruiz b

Rogelio Alejandro Alonso Morales c

Luis E. Eguiarte d

Felipe de Jesús Ruiz-López b*

a
Universidad Nacional Autónoma de México. Posgrado en Ciencias Biológicas, Circuito de
Posgrados, Edificio D, 1er Piso, Ciudad Universitaria, Alcaldía Coyoacán. 04510, Ciudad de
México, México.
b
Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias. Centro Nacional de
Investigación Disciplinaria en Fisiología y Mejoramiento Animal. Querétaro, México.
c
Universidad Nacional Autónoma de México. Facultad de Medicina Veterinaria y Zootecnia.
Ciudad de México, México.
d
Universidad Nacional Autónoma de México. Instituto de Ecología, Departamento de
Ecología Evolutiva. Ciudad de México, México.

*Corresponding author: ruiz.felipe@inifap.gob.mx

Abstract:

The objective was to calculate the parameters, correlations and describe the genetic trends of
reproductive traits in Holstein cattle from Mexico in three different periods of time, in
females born between 2006 and 2019, using records of the Mexican Holstein association.
The reproductive parameters calculated were: calving to first insemination interval (CFI),
number of services per conception (NSC), days open (DO), interval between calving (IBC),

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and age at first calving (AFC). The components of variance were estimated using restricted
maximum likelihood in an animal model, to calculate genetic parameters (narrow-sense
heritability (h2), repeatability (r) and genetic correlations) and phenotypic correlations.
Additive genetic variances and genetic values were also estimated in three periods of time
(P1: 2006-2009, P2: 2010-2013 and P3: 2014-2017). The calculated heritabilities were from
4 to 9 % and the repeatabilities from 8 to 9 %, values close to previous reports for fertility
traits. Phenotypic correlations were positive for almost all reproductive parameters and
genetic correlations were positive over a wide range (0.13-0.99). Analyses by period showed
changes, possibly derived from the influence of the import and use of germplasm from
foreign bulls (mainly from the United States of America and Canada) which implement
genomic selection and include fertility traits. The present research has made it possible to
update the information on reproductive and genetic parameters in fertility traits that can be
incorporated into national genetic evaluations.

Keywords: Reproductive traits, Holstein cattle, Genetic parameters, Heritability,

Repeatability.

Received: 30/05/2022

Accepted: 23/01/2023

Introduction

In domestic animals such as cattle, the study of reproduction is based on records of the
reproductive events of herds, including dates of birth, insemination and calving, with which
it is possible to calculate reproductive parameters (RPs)(1,2). Examples of RPs are the days
from calving to first service/insemination, the percentage of conception at first service, the
interval between calvings, the days from calving to conception, the number of services per
conception and the age at first calving, which may be correlated with each other(3,4). The
reproductive parameters that are used for the study of cow fertility differ between
countries(3,5).

In dairy cattle of the Holstein breed, there are several reports published worldwide on a
reduction in fertility from the 80s to the early 2000s(6,7), apparently due to the genetic
improvement of milk production(7,8), which is related to a negative genetic correlation
reported between fertility and milk production(9,10).

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For dairy farmers, a reduction in fertility causes significant losses and economic
impacts(10,11). By requiring a greater number of inseminations for a cow to become pregnant,
insemination costs increase and the interval between calvings lengthens(7,9), decreasing the
average milk production of the herd, as well as the number of calves per cow(10,11). Likewise,
fewer cows will become pregnant within the required period, which increases the
“involuntary disposal” or slaughter of the cow due to low fertility(10,11), increasing costs due
to an additional replacement animal is bought or raised in the farm(11,12).

Therefore, the fertility and the beginning of the productive life of the cow are priority factors
in the lifetime productivity of dairy cattle(2,12). Their study is important for the future
profitability of production systems and reduction of slaughter generated by deficient
fertilities(9,11).

To increase milk production in cattle, genetic selection has been used, with methodologies
developed and implemented from phenotypic and genealogical records(13,14). For a little over
20 yr, the use of genome-wide genetic markers, such as single-nucleotide polymorphisms
(SNPs), has been proposed to aid selection, through genomic selection (GS)(15), first used in
Holstein cattle from the United States in late 2009(16).

For several generations in countries such as the United States of America and Canada,
fertility traits in Holstein cattle have been evaluated and improved(16,17), and the use of GS
resulted in an increase in the rate of improvement, despite being generally low heritable
traits(16,17) and the possible impact that this selection has had on the genetic parameters and
values of populations that import germplasm from these countries, such as Mexico, is not
known. On the other hand, in Mexico there are few works with the aim of studying genetic
aspects of fertility traits(18,19,20), despite their high economic and functional impact on
Mexican herds.

Therefore, the objective of the present study was to calculate the parameters, correlations and
describe the genetic trends of reproductive traits in Holstein cattle from Mexico in three
different periods of time.

Material and methods

Data and reproductive parameters analyzed

This study included 415,859 reproductive records of Holstein cows (Bos taurus taurus) that
had information on milk production, insemination events (artificial (AI) or natural mounting

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(NM)) and calving, occurred between January 2006 and December 2019; information
provided by the Holstein association of Mexico.

The RPs calculated and included in the present study were: calving to first insemination
interval (CFI), measured as the days elapsed between calving and the first
service/insemination; number of services per conception (NSC), it corresponds to the number
of services/inseminations until gestation; days open (DO), are the days elapsed between the
calving and the new gestation of the cow, the interval between calvings (IBC), measured as
the number of months between two consecutive calvings, and finally, the age at first calving
(AFC), which corresponds to the recorded age on which each cow has its first calving. The
first four RPs together indicate the ability of cows to conceive and restore the estrous cycle;
and the ability of farmers to detect estrus and insemination(5). The last RP (age at first calving)
is a measure of the female calf’s ability to grow and conceive at an early age.

To ensure the quality of the data of the RPs studied, biologically feasible limits were applied
to prevent the use of misinformation (outliers), considering the values reported by the
International Bull Evaluation Service (INTERBULL)(21). Thus, the following records were
excluded: days from calving to first service < 20 or > 365 d, when the number of services per
conception was greater than 10, when the days open were <20 or > 365, when the intervals
between calving were outside the range of 290 to 762 d and when the age at first calving was
<18 or > 40 mo, the descriptive statistics of the variables studied are shown in Table 1. After
quality control, the dataset consisted of 202,545 records for CFI and NSC; 194,816 records
for DO; 139,901 for IBC and 103,467 for AFC. The pedigree files included between 103,467
and 202,545 animals, depending on the trait evaluated and included up to three generations
of ancestors of animals with reproductive records.

Estimation of variance components

The variance components for CFI, NSC, DO and IBC were estimated with a repeatability
animal model, separately for each characteristic to take advantage of as much information as
possible. In the animal model, the availability of individual records allows the prediction of
genetic values for all individuals in the population (even if they do not have phenotypic
information but genealogical information). The genetic values are calculated with the best
linear unbiased predictor, with a mixed equation model (equation 1)(13), where the random
effect refers to the animal that has records and its ancestors, and the fixed effects are all those
environmental effects, such as the year of birth(14).

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In order to control sources of variation other than those studied in the models of the RPs, the
following were included as fixed effects: the subclasses of herd/year/season of calving or
herd-year season of birth for AFC, with four seasons defined according to the month of
calving (January to March, April to June, July to September and October to December); the
level of production (classified into four levels for all farms from 1 to 4, seeking to assign
cows to the quartile that describes their level of production within their herd/year/season of
calving, with 1 being the one with the lowest production and 4 being the one with the highest
production); the number of calving (grouping cows with four lactations or more in a single
class) and age at calving. The animal and the cow’s permanent environment were included
as random effects. In the case of AFC, there was no permanent environment.

The general animal model used was:

y= Xb + Zu + e [equation 1]

Where:
y=variable of interest (vector of records of: CFI, NSC, DO, IBC, AFC);
b= vector of fixed effects (herd-year-season of calving (hys) or herd-year season of birth
(hysb) for AFC, production level (pl), number of calving (noc) and age at calving (ac));
u= vector of random effects (effect of animal and permanent environment when applicable);
X= incidence matrix that relates observations to fixed effects;
Z= incidence matrix that relates observations to random effects (animal and permanent
environment);
e= vector of the effects of the error or residuals.

No transformations of the studied variables were performed because previous studies(12)

showed that they are not necessary. Estimates of variance components were made by means
of restricted maximum likelihood(22), using the BLUPF90 software (23).

Multivariate analyses

In order to estimate the genetic covariances and correlations for each pair of traits, bivariate
analyses were performed using the models described above.

The general model used was:

𝑦1 𝑋 0 𝑏1 𝑍 0 𝑢1 𝑒1
[𝑦 ] = [ 1 ] [ ]+[ 1 ] [𝑢 ] + [𝑒 ]
2 0 𝑋2 𝑏2 0 𝑍2 2 2
where subindices 1 and 2 identify the pair of traits to be evaluated (CFI, NSC, DO, IBC,
AFC) with their respective fixed and random effects.

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Genetic parameters and correlations

Narrow-sense heritabilities (h2), repeatability (r) and genetic correlations(24) between the RPs
were obtained based on the estimates of the variance components using the BLUPF90
software (23). Standard deviations of the estimated parameters were approximated based on
their expectations using the REMLF90 software (23).

Analysis by periods (additive genetic variance and genetic values)

Taking into consideration that the import of germplasm from different countries worldwide,
especially from the United States and Canada, has been increasing in recent years,
representing almost 80 % of the bulls used in the population studied, as well as changes in
the values of the reproductive parameters of these countries (due to the introduction of
genomic information in the evaluation and selection processes since 2009), data were
grouped into three periods, each of four years (P1: 2006-2009, P2: 2010-2013 and P3: 2014-
2017). In the first period, selection was considered to be based on pedigree, which uses
information from the records of cows and their relatives. In the second period, the
incorporation of genomic information into the selection began. In the third period, it is
considered that genomic selection is established, and with it the possibility of increasing the
accuracy of evaluations of low heritability traits.

Results and discussion

Reproductive parameters

The average for CFI was 71.5 d, which could indicate that the first inseminations are being
carried out between 10 and 30 d after the restoration of the estrous cycle (21 d) and the
subsequent voluntary waiting period (usually 40 d). In comparison, measurements made in
different countries reported averages ranging from 70 to 93 d (Table 1).

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Table 1: Descriptive statistics of reproductive parameters in Holstein populations,

including those of the present study

CFI NSC DO IBC AFC

Author Country
± SD ± SD ± SD ± SD ± SD
29 USA 87.6 ± 26 - 109.2 ± 38 - 33.6 ± 12.1
6 USA - - 131.5 - -
26 CAN 80.2 1.66 108.4 - -
34 USA - - - - 27.2 ± 3.3
36 USA 70.5 ± 26 - - - -
36 USA 91.5 ± 40 - - - -
31 ITA 84.5 ± 37 1.7 ± 0.1 - 413.5 ± 82 -
12 ESP 81 ± 28 1.9 ± 1.2 117 ± 57.1 400 ± 60 -
33 MEX - - - 406 ± 67 27.6 ± 2.9
33 USA - - - 401 ± 62 27.1 ± 3.1
28 DNK 81.3 ± 40 2.2 ± 1.5 133.3 ± 76 413.1 ± 76 -
18 MEX 73 ± 45 2.0 ± 1.3 101 ± 54 389 ± 57 -
25 TUN 93.2 ± 80 2.6 ± 1.7 150.9 ± 76 444.2 ± 102 -
19 MEX - - - 418.6 ± 89 -
30 IRN 72.9 ± 35 2.1 ± 1.4 117.7 ± 64 393.9 ± 63 -
27 COL - 1.6 ± 1.0 127.2 ± 77 410.3 ± 78 -
32 CZE - - - 400 ± 59 25.3 ± 2.5
Present study MEX 71.5 ± 23 2.7 ± 1.9 131.6 ± 74 413.6 ± 85 24.2 ± 2.7
CFI = calving to first insemination interval in days, NSC= number of services per conception, DO = days
open, IBC= interval between calvings in days, AFC= age at first calving in months, = average for the traits,
SD= standard deviation

The results for NSC were 2.7 on average (Table 1), similar to the 2.55 reported in a population
of this same breed (Holstein) from Tunisia(25). Lower averages from 1.58 and up to 2 services
per conception have been reported in Holstein populations of Spain(12), Mexico(18), Canada(26)
and Colombia(27). The difference can be caused by the upper limits of 7 services established
by the aforementioned authors, with respect to the 10 services considered in the estimates of
this work.

For DO, the results showed 131.6 d, in accordance with the 131.5 d reported in a population
of the United States(6), of Denmark with 133.3 d(28); or in Colombia with 127.2 d(27) (Table
1). While lower averages were obtained in other studies from 101 d to 117 d(18,29,30). In
contrast, a value of 150 d was reported in Tunisia(25). As in the traits discussed above, the
differences between this study and those cited here could be due to the variation of the limits

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used for each population, as well as to postpartum health problems specific to each population
studied(29).

The average of IBC of 413 d coincides with other populations(28,31) (Table 1), as well as
reports ranging from 400 to 416 d(19,27,32). On the other hand, lower averages of 389 d were
reported in 2010 in a Holstein and Brown Swiss Mexican population under subtropical
conditions(18).

For AFC, the results were 24.15 mo (Table 1), similar to that reported in several studies, such
as the 25.34 mo mentioned in the Czech Republic(32), 27.1 in populations of the United
States(33,34), as well as 27.6 mo in another Mexican population(33) of the Holstein association
of Mexico that includes data from 1971 to 1995. In contrast, 33.6 mo were reported in a
population of the United States(29). This parameter shows about the female calf’s ability to
grow and conceive at an early age(5), which may be different for each population, since there
is a cost-benefit ratio in achieving earlier ages at first calving(11). That is, it is a voluntary and
economic decision of the administrators of each population, because the age at first
insemination determines AFC and the first is almost always an administrative decision, since
the cost associated with developing the female calf faster can be important in some
environments (countries and production systems)(11,35).

The differences obtained in the RPs evaluated and those mentioned above illustrate the
difficulty in estimating them and the lack of consensus on acceptable biological limits in the
measurements of these traits(3,5), which generates possible biases to the estimates and
difficulties for comparison. Other issues such as the dependence of these traits on the
conditions of each country(26,27,30), the economic purposes of each farm(12), as well as the
conditions and management that vary from one herd to another(35), may be generating the
changes observed between countries.

Genetic parameters

Heritability ranged from 4 % to 9 % for the RPs studied, being higher for AFC (9 %), of 5 %
for NSC, DO and IBC, and the lowest for CFI with 4 % (Table 2), similar to that previously
reported for fertility traits(12,26,27).

The lowest heritability obtained was for CFI, with 4 %, similar to the 3 % reported in
populations of Tunisia(25) and Canada(26), as well as 5 % of a Spanish population(12). However,
the results of the present study were lower than the 6 % reported by Italy(31) and Iran(30), as
well as the 9 % estimated in Denmark(28).

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Table 2: Narrow-sense heritability and repeatability in Holstein populations, including

those of the present study

CFI NSC DO IBC AFC

Author Country
h2 r h2 r h2 r h2 r h2

6 USA - - - - 0.05 - - - 0.05

26 CAN 0.03 0.08 0.03 0.07 0.05 0.10 - - -
31 ITA 0.06 0.14 0.03 - - - 0.07 - -
12 ESP 0.05 - 0.02 - 0.04 - 0.04 - -
28 DNK 0.09 - 0.03 - 0.07 - 0.07 - -
25 TUN 0.03 0.13 0.03 0.03 0.04 0.14 0.06 0.15 -
19 MEX - - - - - - 0.01 - 0.03
30 IRN 0.06 - 0.05 - 0.08 - 0.07 - -
27 COL - - 0.04 0.07 0.08 0.1 0.09 0.09 -
32 CZE - - - - - - 0.03 0.09 0.03
0.04 0.09 0.05 0.08 0.05 0.08 0.05 0.08 0.09
PS MEX
(0.003) (0.003) (0.004) (0.003) (0.004) (0.003) (0.004) (0.003) (0.007)
RP: reproductive parameter, CFI: calving to first insemination interval in days, NSC: number of services per conception,
DO: days open, IBC: interval between calvings in days, AFC: age at first calving in months, h2: heritability, r:
repeatability, standard error in parentheses, PS: present study.

The estimated heritability for NSC was 5 %, consistent with that reported in an Iranian
population(30), in a Colombian(27) with 4 %, and the 3 % reported in populations of Spain(12),
Tunisia(25) and Canada(26).

Regarding DO, the heritability is consistent with that reported in Canada(26) and the United
States(6) of 5 %, as well as those estimated in Spain(12) and Tunisia(25) with 4 %; on the other
hand, 7 % was reported in Denmark(28) and 8 % estimated in populations of Colombia and
Iran(27,30).

For IBC, a heritability of 5 % was estimated, which was also similar to previous works done
in Tunisia (6 %)(25), Spain (4 %)(12), Czech Republic (3 %)(32) and lower than the 7 % in
Italy(31), Denmark(28) and Iran(30). The maximum heritability value reported for this
reproductive parameter is 9 % in Colombia(27), and the lowest value reported is 1 %, in the
same study population over a period of time from 1998 to 2003(19).

The highest heritability obtained in the present study was for AFC, with 9 %, higher than the
5 % reported in one population of the United States(6), or the 3 % reported in another Mexican
population(19) and in the Czech Republic(32). Therefore, it would be expected that, in our
population, the response to selection for this trait would have a better result, due to
differences in heritabilities.

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The repeatability estimate for NSC and DO was the same, slightly higher for CFI and IBC
with 9 % (Table 2). Other values reported for the Holstein breed are similar to those obtained
in the present study(26,27,32).

The genetic parameters of the reproductive traits included in this study have relatively low
values (heritability < 10 % and repeatability < 15 %)(12,19,26), so, in addition to genetics, the
environment has an important effect as a source of variation(36). That is, other factors such as
health(29,36), nutritional aspects(36), as well as administrative management issues (voluntary
waiting period) by producers impact these traits(6,26,29).

It is important to highlight that, although the heritabilities found in this study are low, genetic
improvement can allow positive, permanent and cumulative benefits to be obtained in
populations(11,16,17).

The estimated phenotypic and genetic correlations were positive for almost all reproductive
parameters (Table 3). The highest genetic correlations were found between DO and IBC
(0.99), as well as high values of 0.84 and 0.88 for NSC with DO and IBC, respectively. For
CFI with DO and IBC, an intermediate correlation (0.55 and 0.51) was found. For the other
combinations of fertility traits, low correlations (from 0.09 to 0.22) were found.

Table 3: Genetic correlations above the diagonal (standard error), phenotypic correlations
below the diagonal (number of records for each pair of traits)
CFI NSC DO IBC AFC
CFI - 0.13 (0.06) 0.55 (0.05) 0.51 (0.06) 0.09 (0.07)
NSC -0.05 (202,545) - 0.84 (0.02) 0.88 (0.02) 0.22 (0.08)
DO 0.26 (192,527) 0.77 - 0.99 (0.002) 0.09 (0.08)
(194,816)
IBC 0.23 (138,277) 0.78 0.98 - 0.12 (0.11)
(139,901) (137,158)
AFC 0.08 (103,467) 0.01 (84,675) 0.06 (80,467) 0.07 (62,999) -
CFI = calving to first insemination interval in days; NSC= number of services per conception; DO = days
open; IBC= interval between calvings in days; AFC= age at first calving in months.
All phenotypic correlations had a P<0.001.

In particular, the lowest estimated genetic correlation was for the association between CFI
and NSC (0.13), similar to the 0.12 reported for these parameters in an Italian population(30),
and it was also the lowest estimated for another Mexican population(18) with a value of 0.25.
CFI and DO had a positive and moderate genetic association of 0.55, in contrast, in another
population of Spain(12) and Mexico(18), a value of 0.82 was obtained for this association. The
correlation estimator for CFI and IBC indicates that these two reproductive traits show a

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positive and moderate genetic association (0.51), and lower than another Mexican population
(0.89)(18).

The correlation between NSC and DO was positive and high (0.84), although lower than that
reported in Spain(12), Mexico(18) and Colombia(27) (0.94, 0.97, 0.98, respectively). A similar
correlation value of 0.88 was found between NSC and IBC, close to the 0.89 reported in
Spain(12), lower than the 0.95 and 0.97 in Colombia(27) and Mexico(18), respectively; unlike
the 0.61 reported in Italy(30). High values of genetic correlation indicate the possibility of
selecting for a trait, because as one can be decreased or increased, the same effect on the
other is expected. In addition, the close relationship and dependence of the estrus detection
rate and pregnancy between these measurements may explain these results, as well as the fact
that the same genetic regions are encoding these traits(29,37).

Between DO and IBC, high genetic correlation values (0.99) were found, identical to those
reported in Spain(12) and Iran(30) and similar to the 0.98 reported in Colombia(27). The results
found are to be expected, because the IBC includes DO and the length of gestation(29), as a
consequence, any variation of days open should result in changes in the interval between
calvings. Additionally, these two traits, being almost equivalent, suggest that they are
controlled by the same genes or genetic regions, which is known as the pleiotropic effect(37).

The age at first calving is influenced by the environment and management practices common
to female calves and heifers born in the same period (year or year-season)(6), which was
corroborated by the low correlations found between it and the other RPs (Table 3), so these
results suggest genetic and phenotypic independence among them.

Analysis by periods

Additive genetic variance

The results of the estimates of variances by period can be divided into two groups of
comparisons P1 vs P2 and P2 vs P3 (Table 4). In P1 vs P2, for additive genetic variances,
was observed similar values in CFI (12.48-11.17), NSC (0.15-0.16), DO (229.8-214.6) and
IBC (478-466.7), except for AFC (1.23-0.19) which showed a significant decrease, possibly
due to the selection on milk production during this period, because as a cow becomes
pregnant, it will begin its productive life.

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Table 4: Additive genetic variance and heritability for five reproductive parameters in three
periods of time (P1: 2006-2009, P2: 2010-2013 and P3: 2014-2017)

Reproductive Number of 2
Period σ a ± SE h2
parameter records
P1:06-09 71,667 12.48 ± 2.2 0.03
CFI P2:10-13 63,952 11.17 ± 2.0 0.03
P3:14-17 43,233 19.88 ± 3.0 0.06
P1:06-09 71,667 0.15 ± 0.02 0.05
NSC P2:10-13 63,952 0.16 ± 0.02 0.05
P3:14-17 43,233 0.10 ± 0.02 0.04
P1:06-09 69,417 229.8 ± 35.32 0.04
DO P2:10-13 62,783 214.6 ± 33.87 0.04
P3:14-17 42,790 194.8 ± 25.66 0.05
P1:06-09 54,005 478.0 ± 51.1 0.07
IBC P2:10-13 48,388 466.7 ± 52.5 0.07
P3:14-17 25,801 266.8 ± 53.9 0.05
P1:06-09 31,542 1.23 ± 0.12 0.22
AFC P2:10-13 34,094 0.19 ± 0.04 0.04
P3:14-17 31,222 0.15 ± 0.03 0.04
CFI = calving to first insemination interval in days, NSC= number of services per conception, DO = days
open, IBC= interval between calving in days, AFC= age at first calving in months, σ 2a= additive genetic
variance, SE= standard error, h2= narrow-sense heritability.

In P2 vs P3, an increase in CFI (11.17-19.88) was observed in additive genetic variance,

which suggests genetic diversity and the larger it is, the greater the opportunity for response
to selection(38). In contrast, decreases were observed for NSC (0.16-0.10), DO (214.6-194.8)
and IBC (466.7-266.8), while AFC was similar to the previous period (0.19-0.15). The
decrease in the additive genetic variances of NSC, DO and IBC may be due to a selective
pressure of these traits.

Genetic values

The changes over time between groups P1 vs P2, and P2 vs P3 are shown in Figures 1a and
1b, where it can be seen how, in general, genetic trends were similar between P1 and P2,
decreasing in P3. These results are in line with the farmers’ criteria for choosing semen, who,
during the first two periods, emphasized the volume of milk, which decreased in P3, while
in case of reproductive traits, there were no changes in P1 and P2, but a positive change was
observed in P3 [Durán C., 2022, unpublished data].

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Figure 1: Genetic trends for CFI (=IPPS), NSC and AFC (=EPP) (a) and for DO (=DAB)
and IBC (=IEP) (b) in three periods of time (P1: 2006-2009, P2: 2010-2013 and P3: 2014-
2017)

For group II (P2 vs P3), a decrease in average genetic values can be observed for CFI, DO,
IBC and AFC, but not for NSC. This reduction in genetic values is desirable because what is
sought in these fertility traits studied is the decrease.

These changes in fertility traits could possibly be attributed to the import and use of
germplasm from the United States since 1950(39), which has improved fertility traits since
2003(17) and uses genomic selection since 2009(16).

For NSC, no changes were observed in genetic values, probably because this characteristic
depends significantly on insemination techniques, which do not depend on the population of
origin of the germplasm.

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The results of this study show how the incorporation of genomic information in the selection
processes of the countries of origin of imported germplasm has apparently had a positive
effect on the population (Figures 1a and 1b), regardless of the low heritability and difficulty
of measuring fertility traits(16,17).

Conclusions and implications

The estimates of heritability and repeatability of the reproductive parameters obtained in the
present study were low and similar to previous reports in the breed. However, the existing
additive genetic variation is sufficient to expect favorable and cumulative (although slow)
results in the selection of these traits. The estimated correlations between the PRs were
positive, highlighting that they were high between NSC, DO and IBC, in particular the
correlation between DO and IBC, with a correlation of 0.99, which suggests that it would be
advisable to select only for one parameter, since it could be considered that its effects are
controlled by the same genetic regions. The results of the estimates of additive genetic
variances and genetic values in different periods show the existence of changes in the
population over time, possibly derived from the implementation of genomic selection in
germplasm supplier countries, such as Canada and the United States.

Acknowledgements and conflict of interest

This paper is part of the requirements for obtaining a Doctoral degree of CDA at the Posgrado
en Ciencias Biológicas, of the Universidad Nacional Autónoma de México (UNAM). Special
thanks to M. Sc. Enoc Mejía, to M. Sc. José Cortés and to MVZ Marisol Morales for their
advice on the management of databases, as well as to the Holstein Association of Mexico for
the information provided. The study was funded by the Instituto Nacional de Investigaciones
Forestales, Agrícolas y Pecuarias with the project No. SIGI: 15352034772. The Consejo
Nacional de Ciencia y Tecnología is also thanked for scholarship no. 631475 provided to
CDA. The authors declare no conflict of interest.

Literature cited:
1. Flaws JA, Spencer TE. Female reproduction, content and volume overview. In: Academic
Press. Encyclopedia of reproduction. 2nd ed. USA: Academic Press Books – Elsevier;
2018: V2:1-2.

552
 Rev Mex Cienc Pecu 2023;14(3):539-555

2. Walsh SW, Williams EJ, Evans ACO. A review of the causes of poor fertility in high milk
producing dairy cows. Anim Reprod Sci 2011;123(3-4):127-138.

3. Banos G. Survey on genetic evaluation procedures for functional traits in cattle in various
countries. Interbull Bulletin 1996;(12).

4. VanRaden PM, Sanders AH, Tooker ME, Miller RH, Norman HD, Kuhn MT, et al.
Development of a national genetic evaluation for cow fertility. J Dairy Sci 2004;87(7):
2285-2292.

5. Jorjani H. International genetic evaluation for female fertility traits. Interbull Bulletin
2006;(34):57.

6. Seykora AJ, McDaniel BT. Heritabilities and correlations of lactation yields and fertility
for Holsteins. J Dairy Sci 1983;66(7):1486-1493.

7. Hageman WHD, Shook GE, Taylor WJ. Reproductive performance in genetic lines
selected for high or average milk yield. J Dairy Sci 1991;74:4366–4376.

8. Peñagaricano F. Genetics and genomics of dairy cattle. In: Academic Press. Animal
Agriculture. Texas, USA: Academic Press Books – Elsevier; 2020:101-119.

9. Van Arendonk JA, Hovenier R, De Boer W. Phenotypic and genetic association between
fertility and production in dairy cows. Livest Prod Sci 1989;21(1):1-12.

10. Vollema AR. Selection for longevity in dairy cattle [doctoral thesis]. Wageningen, The
Netherlands: Wageningen University and Research; 1998.

11. Pritchard T, Coffey M, Mrode R, Wall E. Genetic parameters for production, health,
fertility and longevity traits in dairy cows. Animal 2012;7(1):34-46.

12. González-Recio O, Alenda R. Genetic parameters for female fertility traits and a fertility
index in Spanish dairy cattle. J Dairy Sci 2005;88:3282-3289.

13. Henderson CR. Applications of linear models in animal breeding. Ontario, Canada:
University of Guelph, Guelph; 1984.

14. Mrode RA. Linear models for the prediction of animal breeding values. SRUC UK: Cabi;
2005.

15. Meuwissen TH, Hayes BJ, Goddard ME. Prediction of total genetic value using genome-
wide dense marker maps. Genetics 2001;157:1819–1829.

16. Wiggans GR, Cole JB, Hubbard SM, Sonstegard TS. Genomic selection in dairy cattle:
the USDA experience. Annu Rev Anim Biosci 2017;5:309-327.

553
 Rev Mex Cienc Pecu 2023;14(3):539-555

17. García-Ruiz A, Cole JB, VanRaden PM, Wiggans GR, Ruiz-López FJ, Van Tassell CP.
Changes in genetic selection differentials and generation intervals in US Holstein dairy
cattle as a result of genomic selection. Proc Natl Acad Sci 2016;113(28):E3995-E4004.

18. Ríos-Utrera A, Calderón-Robles R, Rosete-Fernández J, Lagunes-Lagunes J. Análisis

genético de características reproductivas de vacas Holstein criadas en un ambiente
subtropical. Agronomía Mesoamericana 2010;21:245-253.

19. Montaldo HH, Castillo-Juárez H, Valencia-Posadas M, Cienfuegos-Rivas EG, Ruiz-

López FJ. Genetic and environmental parameters for milk production, udder health, and
fertility traits in Mexican Holstein cows. J Dairy Sci 2010;93(5):2168-2175.

20. Montaldo HH, Pelcastre-Cruz A, Castillo-Juárez H, Ruiz-López FJ, Miglior F.

Genotype×environment interaction for fertility and milk yield traits in Canadian,
Mexican and US Holstein cattle. Span J Agric Res 2017;15(2):e0402-e0402.

21. INTERBULL. International Bull Evaluation Service. National genetic evaluation forms
provided by countries. 2020. https://interbull.org/ib/geforms Accessed Sep 1, 2020.

22. Patterson HD, Thompson R. Recovery of interblock information when block sizes are
unequal. Biometrika 1971;58:545-554.

23. Misztal I, Aguilar I, Legarra A, Vitezica Z, Tsuruta S, Lourenco D. Manual for BLUPF90
family of programs. 2014. http://nce.ads.uga.edu/wiki/lib/exe/fetch.php?
media=blupf90_all7.pdf Accessed Nov 1, 2020.

24. Falconer DS. Introduction to quantitative genetics. 2nd ed. New York, USA: Longman
Scientific & Technical; 1981.

25. M’hamdi N, Aloulou R, Brar SK, Bouallegue M, Ben Hamouda M. Phenotypic and
genetic parameters of reproductive traits in Tunisian Holstein cows. Biotechnol Anim
Husb 2010;26:297-307.

26. Hayes JF, Cue RI, Monardes HG. Estimates of repeatability of reproductive measures in
Canadian Holsteins. J Dairy Sci 1992;75(6):1701-1706.

27. Zambrano JC, Echeverri J. Genetic and environmental variance and covariance
parameters for some reproductive traits of Holstein and Jersey cattle in Antioquia
(Colombia). Rev Bras Zootec 2014;43(3):132-139

28. Sun C, Madsen P, Lund M, Zhang Y, Nielsen US, Su G. Improvement in genetic

evaluation of female fertility in dairy cattle using multiple-trait models including dairy
yield traits. J Anim Sci 2010;88:871-878.

554
 Rev Mex Cienc Pecu 2023;14(3):539-555

29. Fonseca FA, Britt JH, McDaniel BT, Wilk JC, Rakes AH. Reproductive traits of
Holsteins and Jerseys. Effects of age, milk yield, and clinical abnormalities on
involution of cervix and uterus, ovulation, estrous cycles, detection of estrus, conception
rate, and days open. J Dairy Sci 1983;66(5):1128-1147.

30. Ghiasi H, Pakdel A, Nejati-Javaremi A, Mehrabani-Yeganeh H, Honarvar M, González-

Recio O, et al. Genetic variance components for female fertility in Iranian Holstein
cows. Livest Sci 2011;139(3):277-280.

31. Biffani S, Canavesi R, Samore AB. Estimates of genetic parameters for fertility traits of
Italian Holstein Friesian cattle. Stocarstvo 2005;59:145-153.

32. Brzáková M, Zavadilová L, Přibyl J, Pešek P, Kašná E, Kranjčevičová A. Estimation of

genetic parameters for female fertility traits in the Czech Holstein population. Czech J
Anim Sci 2019;64(5):199-206.

33. Cienfuegos-Rivas EG, Blake RW, Oltenacu PA, Castillo-Juarez H. Fertility responses of
Mexican Holstein cows to US sire selection. J Dairy Sci 2006;89(7):2755-2760.

34. Castillo-Juarez H, Oltenacu PA, Blake RW, McCulloch CE, Cienfuegos-Rivas EG.
Effect of herd environment on the genetic and phenotypic relationships among milk
yield, conception rate, and somatic cell score in Holstein cattle. J Dairy Sci 2000;83(4):
807-814.

35. Garcia-Peniche TB, Cassell BG, Pearson RE, Misztal I. Comparisons of Holsteins with
Brown Swiss and Jersey cows on the same farm for age at first calving and first calving
interval. J Dairy Sci 2005;88(2):790-796.

36. Weigel KA, Rekaya R. Genetic parameters for reproductive traits of Holstein cattle in
California and Minnesota. J Dairy Sci 2000;83(5):1072-1080.

37. Bulmer MG. The mathematical theory of quantitative genetics. New York, USA:
Clarendon Press, Oxford; 1980.

38. Woolliams JA, Oldenbroek JK. Genetic diversity issues in animal populations in the
genomic era. In Oldenbroek JK editor. Genomic management of animal genetic
diversity. Wageningen, The Netherlands: Wageningen Academic Publishers; 2017:13-
45.

39. Cienfuegos-Rivas EG, Oltenacu PA, Blake RW, Schwager SJ, Castillo-Juarez H, Ruiz
FJ. Interaction between milk yield of Holstein cows in Mexico and the United States. J
Dairy Sci 1999;82(10):2218-2223.

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Article

Additive and non-additive genetic effects for reproductive traits in a

Holstein- Brown Swiss diallel in humid subtropical climate

Luis Adrián García Bravo a

Vicente Eliezer Vega-Murillo a*

José Alfredo Villagómez- Cortes a

Juan Prisciliano Zárate-Martínez b

Otto Raúl Leyva-Ovalle c

René Carlos Calderón Robles d

Ángel Ríos-Utrera a

Moisés Montaño Bermúdez e

Guillermo Martínez-Velázquez f

a
Universidad Veracruzana. Facultad de Medicina Veterinaria y Zootecnia, 91710, Veracruz,
Ver. México.
b
Instituto Nacional de Investigaciones Forestales Agrícolas y Pecuarias (INIFAP). Campo
Experimental La Posta, Veracruz, México.
c
Universidad Veracruzana. Facultad de Ciencias Biológicas y Agropecuarias. Veracruz, Ver.
México.
d
INIFAP, Sitio Experimental Las Margaritas, Puebla, México.
e
INIFAP, CENID Fisiología y Mejoramiento Animal, Querétaro, México.
f
INIFAP, Campo Experimental Santiago Ixcuintla, México.

*Corresponding author: vvega@uv.mx

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Abstract:

Crossbreeding allows taking advantage of additive genetic differences between breeds, they
also allow making use of heterosis and complementarity. Therefore, it is necessary to
generate information on the efficacy of crosses compared to pure breeds under the conditions
of interest. The objective was to quantify the impact of additive and non-additive genetic
effects for days to first estrus (DFE), days to first service (DFS), days open (DO), services
per conception (SPC), calving interval (CI) and gestation length (GL). The productive and
genealogical information of females from a diallel between Holstein (HO) and Brown Swiss
(BS), a total of 148 cows of the breeds HO (n=43), BS (n=64) and their reciprocal crosses
HO-BS (n=20) and BS-HO (n=21), was used. Contrasts were used to estimate individual
heterosis and differences between direct genetic effects and between maternal genetic effects
based on Dickerson models. The results showed that heterosis and differences between
maternal effects were not significant (P>0.05) for any of the traits studied. Differences
between direct genetic effects were only important (P<0.05) for SPC and GL. In conclusion,
the heterosis generated by the crossbreeding between HO and BS did not influence the
reproductive efficiency of females. Maternal effects were not different between HO and BS.
Direct genetic effects for SPC and GL favored the BS breed.

Keywords: Bovine, Diallel crossbreeding, Direct genetic effect, Heterosis, Statistical

models, Linear models, Reproductive parameters, Reproductive traits.

Received: 27/05/2022

Accepted: 06/03/2023

Introduction

The selection of purebred animals allows improving traits of zootechnical importance, such
as milk production in dairy cattle(1,2). And precisely the high pressure of selection on this
phenotype has generated antagonistic genetic correlations with longevity(3), udder health(4)
and fertility(5). Generating deteriorations(6,7,8) observable in a low general productive
performance(9). Establishing a system of crossbreeding between breeds is an alternative to
solve this problem(10).

The purpose of crossbreeding between genetically distant individuals is to generate new

combinations of genes that allow taking advantage of direct (average deviation caused by the
direct effect of the genes of the purebred individual) and maternal (average deviation due to

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the effects of the dam’s genes through the maternal environment) additive genetic differences
between breeds, and mainly to use non-additive genetic effects due to dominance and
epistasis, such as heterosis(10-14), which allows the average of hybrid offspring to be
phenotypically superior to the average of contemporary parental pure breeds(15,16).
Individuals from crosses may have advantages over their parents if the traits of interest show
positive or useful heterosis(17), which is reflected in better reproductive rates(18).

On the other hand, in the tropical conditions of Mexico the main interest is milk production,
for this reason, crosses are made with Brown Swiss or Holstein cattle(19), but not among them.
The crossbreeding of these breeds could allow preserving the dairy ability and improving the
general reproductive indexes. Therefore, evaluating the phenotypic behavior of the hybrid
progeny based on the environment and the productive conditions of interest will provide the
necessary knowledge when deciding the mating strategies that will be used to maximize the
estimators of the reproductive parameters. The objective of the present study was to quantify
the additive and non-additive genetic effects on the reproductive efficiency of Holstein,
Brown Swiss females and their reciprocal crosses in a subtropical environment.

Material and methods

Geographical conditions

The study was conducted at the “Las Margaritas” experimental site of the National Institute
of Forestry, Agricultural and Livestock Research (INIFAP, for its acronym in Spanish),
located in the municipality of Hueytamalco (latitude 19°, 20'N and longitude 97°, 20'W), in
the northeastern sierra of the state of Puebla, Mexico, at 450 m asl. The climate is classified
as humid subtropical Af(c)(20) and the average annual temperature is 20.8 °C, ranging from
15.3 (winter) to 24.2 °C (summer), and the average annual rainfall is 1,270 mm. The abundant
rainy season occurs from July to October; drizzle with temperature drop occurs from
November to February(21).

Herd characteristics

The information used was from reproductive and genealogical records of 148 Holstein (HO;
n=43) and Brown Swiss (BS; n=64) cows and their reciprocal crosses HO-BS (n=20) and
BS-HO (n=21), which were produced with 85 sires and 121 dams through artificial

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insemination and natural mating, and which were born between 1997 and 2006 and calved
between 1998 and 2014.

Animals and reproductive management

Heifers weighing 350 kg or more and presenting no abnormalities in the reproductive tract
after transrectal palpation were included. The detection of estrus was carried out from 0600
to 0700 h and from 1700 to 1800 h, with the help of a bull with a chin-ball marker. Females
in estrus were inseminated with the conventional technique 12 h after visual detection of
estrus. The diagnosis of gestation was made by transrectal palpation 45 d after the last service.

Health

The cows were vaccinated against clostridiosis, pasteurellosis (twice a year; March and
September) and bovine paralytic rabies (September). Tests for the control of brucellosis and
tuberculosis were also performed every 14 mo (free herd). To control ectoparasites (ticks and
flies), immersion bath was performed every 14 to 30 d, depending on the degree of infestation
and the ectoparasiticidal product used. Dried cows were dewormed one month before
calving, in case the coproparasitoscopic diagnosis indicated it.

Feeding management

The cows were kept in an intensive rotational grazing system, mainly in African star grass
(Cynodon plectostachyus) and pangola grass (Digitaria decumbens). The periods of
occupation and rest of the paddocks were from 2 to 3 d and from 35 to 40 d respectively,
depending on the season of year, with a stocking rate of 2.5 animal units per hectare per year.
During the dry season (November to March), the cows received 20 to 30 kg/animal/d of
chopped fresh fodder, mainly Japanese cane (Saccharum sinense) or sugarcane (Saccharum
officinarum), added with a mixture of molasses (97 %) and urea (3 %) at a rate of two to three
kilograms per animal per day. In addition, lactating cows received 3.5 kg/d of a commercial
concentrated feed (16 % crude protein and 70 % total digestible nutrients) during milking,
while dry cows were fed 2 kg/d of the same type of feed. Together, they were provided with
a mixture of minerals and water ad libitum.

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Milking management

The cows were separated from their calves on the third day postpartum, then classified and
managed in three batches: 1) cows from calving to fifth month of lactation, 2) cows from the
fifth month of lactation to drying and 3) dry cows. Milking of cows began on the fourth day
after calving. They were milked twice a day (0600-0800 and 1400-1600 h) with a milking
machine (Alfa Laval Agri, In-churn, United Kingdom). The cows were dry off at seven
months of gestation or when their milk production was less than 2 kg per day. The averages
of lactation length (days) and production per lactation (kg) were for BS: 323 ± 65 and 3190.55
± 88, HO: 324 ± 69 and 3685.65 ± 1092, BS-HO: 323 ± 61 and 3887.29 ± 1058, and HO-BS
328 ± 59 and 3910.44 ± 1315, respectively.

Variables analyzed

The variables analyzed were: 1) days to first estrus (DFE), defined as the days that elapsed
from the last calving to the first manifest estrus; 2) days to first service (DFS), defined as the
days that elapsed from the last calving to the first service of artificial insemination or natural
mating; 3) days open (DO), defined as the period between the last calving and the next
conception; 4) services per conception (SPC), defined as the number of inseminations or
natural mating necessary to achieve gestation; 5) calving interval (CI), defined as the days
elapsed between two consecutive calvings and 6) gestation length (GL), defined as the days
that elapse between conception and calving(22).

Data analysis

The MIXED procedure of the SAS(23) statistical package was used. The model included the
fixed effects of year of calving (17 levels: 1998 to 2014), season of calving (three levels:
November-February; March-June and July-October), number of calving (six levels: 1 to 6),
the cows’ breed group (four levels: HO, BS, HO-BS and BS-HO) and the random effect of
the sire nested within the sire’s breed. For the tests of fixed effects, the Satterthwaite
approximation was used for the denominator degrees of freedom.

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Estimation of heterosis

Contrasts were used to estimate individual heterosis and differences between direct genetic
effects and between maternal genetic effects of Brown Swiss and Holstein based on the
following models(24,25):

HO = μn + giHO + gMHO + gNHO

BS = μn + giBS + gMBS + gNBS
HOBS = μn + ½(giHO + giBS) + gMBS + gNBS + hiHOBS
BSHO = μn + ½(giBS + giHO) + gMHO + gNHO + hiBSHO

Where HO and BS = Holstein and Brown Swiss; HOBS and BSHO = reciprocal crosses
between HO and BS; μn = average of the pure breeds involved in the diallel cross; giHO and
giBS = deviation due to the average direct effect of the individual’s genes, which come from
the HO or BS breed; gMHO and gMBS = deviation due to the average effects, through the
maternal environment, due to genes of dams of the HO or BS breed; gNHO and gNBS =
deviation due to average effects, through the maternal environment of HO or BS granddams,
which may affect the maternal ability of their daughters; hiBSHO and hiHOBS = deviation due to
the increase in average heterozygosity of the F1 crosses BS-HO and HO-BS. To estimate the
differences between the direct genetic effects of HO and BS, the contrast (HO + HOBS - BS
- BSHO) was used, while the differences between maternal genetic effects were estimated
with the contrast BSHO - HOBS, assuming that gNHO - gNBS was equal to zero. Individual
heterosis was calculated by the contrast [HOBS + BSHO - HO - BS] / 2.

Results

Table 1 shows the significance levels for the fixed effects considered in the models for each
trait analyzed.

Table 1: Levels of statistical significance of the fixed effects considered in the statistical
model for analyzing reproductive traits
Effects DFE DFS DO SPC CI GL
Cow’s breed group 0.298 0.216 0.738 0.008 0.364 <0.0001
Year of calving 0.001 0.001 0.003 0.079 0.019 0.233
Season of calving 0.737 0.859 0.286 0.192 0.050 0.214
Number of calving 0.739 0.770 0.338 0.781 - 0.560
DFE= days to first estrus, DFS= days to first service, DO= days open, SPC= services per conception, CI=
calving interval, GL= gestation length.

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The effect of the cow’s breed group was significant for SPC (P=0.008) and GL (P=0.001).
In the case of year of calving, it was significant for DFE (P=0.001), DFS (P=0.001), DO
(0.003) and CI (P=0.019). The season and number of calving were not significant (P>0.05)
for any of the traits studied, but they were still included in the model. The least square means
and their standard errors for the traits studied are shown in Table 2.

Table 2: Least square means and standard errors for DFE, DFS, DO, SPC, CI and GL, of
HO, BS cows and their reciprocal crosses (HO-BS, BS-HO)
Cow’s
breed DFE DFS DO SPC CI GL
group
HOBS 97.81±7.18 97.65±7.03 118.78±7.39 1.70±0.12ab 405.83±6.67 281.22±1.00ab
BSHO 94.25±8.15 94.71±7.84 116.05±7.80 1.61±0.11a 406.55±7.11 283.65±1.08a
HO 88.79±6.95 89.19±6.84 124.47±7.41 1.88±0.12b 414.52±6.50 279.84±0.97b
BS 103.31±6.35 104.98±6.23 123.40±6.81 1.45±0.11a 417.74±5.82 287.29±0.88c
DFE= days to first estrus, DFS= days to first service, DO= days open, SPC= services per conception, CI=
calving interval, GL= gestation length.
a,b,c
Means with different literals within columns are different (P<0.05).

No differences (P>0.05) were found between the pure breeds and their reciprocal crosses in
DFE, DFS, DO and CI. In the case of SPC, HO cows had significantly (P<0.05) higher
number of SPC (1.88 ± 0.12) than the BS-HO (1.61 ± 0.11) and BS ones (1.45 ± 0.11), while
HO-BS cows had an intermediate behavior (1.70 ± 0.12) to parent breeds (P>0.05).
Regarding GL, the BS cows had significantly (P<0.05) longer gestations (283.65 ± 1.08)
than the HO, BS-HO and HO-BS ones, and the HO-BS crosses had an intermediate behavior
with respect to the breeds HO and BSHO (P>0.05). The estimate of heterosis, difference
between direct genetic effects and difference between maternal genetic effects for the traits
under study are shown in Table 3.

Table 3: Estimates of differences between direct genetic effects, difference between

maternal genetic effects and heterosis for reproductive traits
Contrast DFE DFS DO SPC CI GL
Direct -10.95±12.80 -12.86±12.45 3.79±11.54 0.52± 0.17** -3.94 ±11.47 -10.24±1.68**
Maternal 3.56±10.30 2.94±9.96 2.72±9.05 0.09±0.13 -0.72±8.93 -2.43±1.34
Heterosis -0.02±6.75 -0.90±6.55 -6.52±6.00 -0.01±0.09 -9.94±5.75 -0.95±0.87
DFE= days to first estrus, DFS= days to first service, DO= days open, SPC= services per conception, CI=
calving interval, GL= gestation length.
*(P<0.05).

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Direct genetic effects were important (P<0.05) for services per conception and gestation
length. Heterosis and the difference between maternal genetic effects were not significant
(P>0.05) for any of the traits studied.

Discussion

The effect of the cow’s breed group was only significant for SPC and GL (P<0.01). A
possible explanation for the effect of the cow’s breed group on SPC and GL can be attributed
to the fact that highly milk-producing cows tend to have a low conception rate(26). Decreased
fertility in dairy cattle may have a pathological cause and be related to poor nutrition(27). Lack
of adequate nutrition during the prepartum and early postpartum periods can lead to a
reduction in levels of glucose, insulin, insulin-like growth factor 1 and a decrease in the
frequency of LH pulses. At the same time, the levels of beta-hydroxybutyrate, non-esterified
fatty acids and triacylglycerol can increase(28,29). Metabolic disorders predispose cows to
gynecological disorders, thus reducing reproductive efficiency(28,29). On the other hand, it has
been proven that some metabolites present in plasma are also found in the ovarian follicular
microenvironment, and that the single profiles of these metabolites can affect the
reproductive performance of dairy cattle at different physiological stages, such as heifers,
primiparous and multiparous(30). It could also be attributed to differences in the genetic
component resulting from the adaptability of pure breeds to tropical conditions(31). The origin
of the phenomenon is unclear and is described as multifactorial.

The year of calving was significant for DFE, DFS, DO and CI (P<0.01). These results can
be attributed to fluctuations in climatic conditions that influence the physiology of the animal
through adverse or favorable environments(32,33); the season and number of calving were not
significant (P>0.05) for any of the traits studied. These results partially agree with those
obtained by Hundie et al(34) and Niraj et al(35), who also found no differences between seasons
or the number of calvings(36). Contrary to this, in an evaluation of different breeds and crosses
of cattle (Brahman, Indubrasil, BS, BS x Zebu) in southeastern Mexico, it was found that,
during the first and second calving, there are longer periods (CI) due to the stress caused by
lactation, which are erroneously attributed to postpartum management. It is possible that this
stress caused by lactation is not so intolerable in the dairy breeds (HO and BS) used in the
present study(37).

Regarding the least squares means, no differences (P>0.05) were found between the pure
breeds and their reciprocal crosses in DFE, DFS, DO and CI. These results agree with those
obtained by Blöttner et al(2), who found no difference in DO between HO and BS-HO.
Contrary to this, when evaluating some reproductive traits of dairy cows in a rotational

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crossing system using three breeds (HO, Jersey and Montbeliarde) in the highlands of
Mexico, HO cows had more DO and a longer CI than any of the crosses evaluated(38). In
another study, BS, HO-BS and BS-HO cows had significantly less DO(39,40) and the BS-HO
cross a shorter CI than HO cows(39,40,41). Similarly, in a study conducted with databases from
the Canadian Dairy Network (CDN), they evaluated 128,376 reproductive records of 55,648
HO cows, pure and crossed with BS, Jersey, Norwegian Red and Swedish Red bulls, they
found that differences in the number of services and in rates of no return significantly favor
crossbred calves over the pure HO ones, therefore, hybrid animals will avoid the costs
associated with a greater number of services per conception, will reduce the intervals between
calvings and between lactations, which will result in a higher overall production (40). In
addition to the above, the results of the present study show that HO cows require a greater
number of SPC (P<0.05) than the BS-HO and BS ones, while HO-BS cows had an
intermediate behavior. These results partially agree with those of another study where BS
and BS-HO cows had fewer SPC (2.92 and 2.96; respectively) than HO and HO-BS cows
(3.54 and 3.37; respectively)(40). Some authors even indicate that the BS breed and its F1
crosseswith Holstein have better reproductive performance and greater adaptability to
tropical conditions(42). One of the possible causes of the reproductive inefficiency of the
Holstein breed dates back to its historical selection focused on its high milk production(43,44).
The base method for its genetic improvement is the selection of individuals that exceed the
productive merit for this trait(45), but the incorrect and widespread use of assisted reproductive
technologies results in hundreds or thousands of calves from a single parent(46). The strong
pressure in the selection and the poor orientation of the use of reproductive tools have caused
inbreeding problems of consanguinity, which are negatively reflected on reproductive
traits(46,47). Inbreeding generated by mating genetically related animals increases the
frequencies of homozygous genes(48) and restricts heterozygosity(47,49), causing an increase in
the frequency of harmful recessive alleles that manifest as less phenotypic variability(50,51).

Regarding other traits analyzed, Blöttner et al(2) found no differences in the number of SPC
between BS-HO and HO in three consecutive lactations analyzed independently, on the
contrary, in the present study the analysis was carried out considering six lactations together,
it is possibly the explanation of why statistical differences were found between breed groups.
It should be considered that the number of SPC may be influenced by the reproductive health
status of the cows, the management, the type of feeding and the seminal quality of both the
straws and the ejaculate during natural mating(52), however, these factors were not included
in the model due to lack of availability and because all the animals were under the same
climatic and management conditions. BS-HO cows had significantly longer gestations
(P<0.05) (283.65) than the HO (279.84) and BS ones (287.29); HO-BS cows had an
intermediate behavior (281.22). This result agrees with that obtained by Blöttner et al(2), who
reported a longer gestation in BS-HO females (282 d) than in pure Holstein cows (280 d).
Nevertheless, an important factor influencing GL is the sex of the calves(53), unfortunately
these data were not available to be included in the statistical model.

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Heterosis and the difference between maternal genetic effects were not significant (P>0.05)
for any of the traits studied. Conversely, the results in the Mexican highlands where DO, CI
and SPC were evaluated in a rotational crossbreeding with Holstein, Jersey, Montbeliarde
and Swedish Red, found that crosses needed fewer SPC and showed shorther DO and CI than
the HO ones, due to the effect of heterosis(38). This is consistent with results obtained in
subtropical conditions in Egypt(42). Nonetheless, both studies do not mention the heterosis
estimated in the traits analyzed. For example, in BS-HO crosses, 7 % heterosis is reported
for DO (in Mediterranean conditions in the United States)(54), and 1.3, -1.0 and -8.4 %
heterosis for CI, DO and SPC, respectively(40). On the other hand, in the southern region of
Brazil, they evaluated crosses of Simmental cattle (SM) with Holstein (F1, ¾ Holstein x ¼
SM and ¾ SM x ¼ Holstein ) and obtained shorter CI, DO and DFS in F1 and ¾ SM x ¼
Holstein cows(55). The authors mention that crossing HO with SM decreases body weight loss
and guarantees a better postpartum energy balance that benefits the expression of
reproductive traits(55,56). This effect is the result of complementarity between breeds, that is,
combining the dairy ability of the HO with the reproductive ability of the SM cattle (57),
despite the fact that both breeds are Bos taurus. The breeds are complementary to each other
when they excel in different traits and the hybrid calf manifests a desirable performance for
a greater number of phenotypes than pure breeds(57). The hybrid calf manifests superior
phenotypic behavior if the parents contribute genes with dominant effect(18), the new
combinations of different alleles at each locus (heterozygous)(13) will give rise to superior
animals(58). However, for this to occur, it is necessary to use genetically divergent
breeds(59,60). Then, the lack of significance (P>0.05) of the heterosis estimated in the
crossbreeding of the breeds included in the diallel suggests little genetic divergence and
similar gene frequencies between HO and BS for the traits evaluated, which translates into a
scarce or null manifestation of heterosis(11); in this case, some authors recommend increasing
the number of breeds involved in the crosses in order to increase heterozygosity(13,61). On the
other hand, the differences between direct genetic effects were important (P<0.05) for SPC
(0.52; with the HO breed exerting a greater effect) and GL (-10.24; with the effect of the BS
breed predominating).

Conclusions and implications

The heterosis generated by the crossbreeding between HO and BS did not influence the
reproductive efficiency of the females. The estimated heterosis suggests little genetic
divergence between breeds for the traits evaluated. Maternal effects were no different
between Holstein and Brown Swiss. Direct genetic effects favored the BS breed for SPC and
GL. It is recommended to cross with genetically distant breeds to maximize the use of hybrid

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vigor, depending on the production objective and always considering a balance between
productive and reproductive traits.

Acknowledgements

The National Council of Science and Technology is thanked for the grant provided to the
author, with support number 781818.

Conflict of interest

The authors declare that they have no conflict of interest.

Literature cited:
1. Harris BL, Kolver ES. Review of holsteinization on intensive pastoral dairy farming in NZ
J Dairy Sci 2001;84(E.Suppl.):E56–E61. https://doi.org/10.3168/jds.S0022-
0302(01)70197-X.

2. Blöttner S, Heins BJ, Wensch-Dorendorf M, Hansen LB, Swalve HH. Brown Swiss ×
Holstein crossbreds compared with pure Holsteins for calving traits, body weight,
backfat thickness, fertility, and body measurements. J Dairy Sci 2011;94(2):1058–1068.
https://doi.org/10.3168/jds.2010-3305.

3. Hare E, Norman HD, Wright JR. Survival rates and productive herd life of dairy cattle in
the United States. J Dairy Sci 2006;89(9):3713–3720.

4. Seegers H, Fourichon C, Beaudeau F. Production effects related to mastitis and mastitis

economics in dairy cattle herds. Vet Res 2003;34(5):475–491.
doi:10.1051/vetres:2003027.

5. Windig JJ, Calus MPL, Veerkamp RF. Influence of herd environment on health and
fertility and their