Journal of Research in Biology Volume 3 Issue 7

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Panjab University, India

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Fernando Reboredo [Archaeobotany, Forestry, Ecophysiology]
New University of Lisbon, Caparica, Portugal

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Dr. Mrs. Sreeja Lakshmi PV [Biochemistry and Cell Biology]
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University of jan, Spain.

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Dr. Manoranjan chakraborty [Mycology and plant pathology]
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Table of Contents (Volume 3 - Issue 7)
Serial No Accession No Title of the article Page No

1 RA0387 Population density of Indian giant squirrel Ratufa indica centralis (Ryley,
1913) in Satpura National Park, Madhya Pradesh, India.
Raju Lal Gurjar, Amol S. Kumbhar, Jyotirmay Jena, Jaya Kumar Yogesh,
Chittaranjan Dave, Ramesh Pratap Singh and Ashok Mishra.

1086-1092
2 RA0376 Puntius viridis (Cypriniformes, Cyprinidae), a new fish species from
Kerala, India.

Mathews Plamoottil and Nelson P. Abraham.
1093-1104

3

RA0377

A new species of Agathoxylon Hartig from the Sriperumbudur
formation, Tamil Nadu, India.

Kumarasamy D.

1105-1110

4

RA0420

An assessment of bioactive compounds and antioxidants in some
tropical legumes, seeds, fruits and spices.

Dilworth LL, Brown KJ, Wright RJ, Oliver MS and Asemota HN.

1182-1194

5

RA0411

Characterization of silica nanoporous structures of freshwater diatom
frustules.

Dharitri Borgohain and Bhaben Tanti.

1195-1200

6

RA0413

Saprobic status and Bioindicators of the river Sutlej.

Sharma C and Uday Bhan Singh.

1201-1208

Article Citation:
Raju Lal Gurjar, Amol S. Kumbhar, Jyotirmay Jena, Jaya Kumar Yogesh,
Chittaranjan Dave, Ramesh Pratap Singh and Ashok Mishra.
Population density of Indian giant squirrel Ratufa indica centralis (Ryley, 1913) in
Satpura National Park, Madhya Pradesh, India.
Journal of Research in Biology (2013) 3(7): 1086-1092
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Population density of Indian giant squirrel Ratufa indica centralis (Ryley,
1913) in Satpura National Park, Madhya Pradesh, India
Keywords:
Central Indian landscape, Distance sampling, density estimation, Ratufa
indica centralis.
ABSTRACT:

Information on population and distributional status of Indian giant squirrel
Ratufa indica centralis is poorly known from central Indian hills. The species is
endemic to India and widely distributed in Western Ghats, Eastern Ghats and Central
India. In this study using line transect distance sampling we estimated population
density of giant squirrel in Satpura Tiger Reserve (STR), which is a major biosphere
reserve in central India that harbors wide variety of rare endemic and endangered
species. Density estimate with total effort of 276km line transect shows 5.5 ( 0.82)
squirrels/Km
2
. This study provides first baseline information on ecological density
estimate of Ratufa indica centralis in central Indian landscape. Reduction of
anthropogenic pressure should be the first priority for park managers in Satpura Tiger
reserve.
1086-1092| JRB | 2013 | Vol 3 | No 7
This article is governed by the Creative Commons Attribution License (http://creativecommons.org/
licenses/by/2.0), which gives permission for unrestricted use, non-commercial, distribution and
reproduction in all medium, provided the original work is properly cited.
www.jresearchbiology.com
Journal of Research in Biology
An International
Scientific Research Journal
Authors:
Raju Lal Gurjar
1
,
Amol .S. Kumbhar
1*
,
Jyotirmay Jena
1
,
Jaya Kumar Yogesh
1
,
Chittaranjan Dave
1
,
Ramesh Pratap Singh
2
,
Ashok Mishra
2
.

Institution:
1. WWF - India, Nisha
Building, Near Forest
Barrier, Katra, Mandla,
Madhya Pradesh, India.

2. Field Director Office,
Satpura Tiger Reserve,
Hoshangabad, Madhya
Pradesh, India.

Corresponding author:
Amol S. Kumbhar

Email Id:

Web Address:
http://jresearchbiology.com/
documents/RA0387.pdf.
Dates:
Received: 08 Oct 2013 Accepted: 08 Nov 2013 Published: 25 Nov 2013
An International Scientific Research Journal
Original Research

INTRODUCTION
Habitat fragmentation is cited one of the major
reason for the decrease in abundance of arboreal
mammals and isolation of many species into small
population (Umapathy and Kumar, 2000). Indian Giant
Squirrel Ratufa indica centralis is a maroon and buff
colour and is endemic to India with four sub-species. The
conservation status of Indian giant squirrel (IGS) is the
least concern category of IUCN, Appendix II of
CITES and Schedule II (part II) of Indian Wildlife
(Protection) Act 1972 (Molur et al., 2005). Giant
squirrels occur across a wide range of natural forests.
They have been reported from moist deciduous, dry
deciduous and riparian forests (Datta and Goyal, 1996;
Baskaran et al., 2011; Kanoje, 2008; Jathanna
et al., 2008; Srinivas et al., 2008), old mature teak forests
(Ramachandran, 1988) and teak-mixed forests (Kumara
and Singh, 2006). Habitat fragmentation is one of the
major threats which influence giant squirrel population
due to its arboreal nature. Throughout India several
investigators already studied on population status of
Malabar giant squirrel in Western Ghats (Baskaran et al.,
2011; Ramachandran, 1988; Ganesh and Davidar, 1999;
Madhusudan and Karanth, 2002; Kumara and Singh,
2006; Jathanna et al., 2008; Ramesh et al., 2009;
Umapathy and Kumar, 2000). In central India though
there are studies available on ecobiology of Ratufa
indica centralis (Datta, 1993, 1998, 1999; Datta and
Goyal, 1996; Kanoje, 2008; Kumbhar et al., 2012;
Pradhan et al., 2012; Rout and Swain, 2006) but there is
no study available on status and population density of
this species from central Indian landscape.
In the current study we tried to estimate
population densities of Ratufa indica centralis by line
transect distance sampling (Jathanna et al., 2008) in
Satpura Tiger Reserve of central India. It believes that
this kind of effort will help forest department to take
better management and conservation strategies.

MATERIALS AND METHODS
Study area
The Satpura Tiger Reserve (2219 - 22 30N
and 77 56 - 78 20E) covers an area of 1427.87 km
2

(Figure 1) in south east border of Madhya Pradesh state,
it extends from east to west in the southern part of the
district Hoshangabad in Satpura ranges of Central Indian
landscape. The forest types of satpura tiger reserve
consist of southern moist mixed deciduous forest,
southern dry mixed deciduous forest and dry peninsulas
Sal forest (Champion and Seth, 1968). The terrain of
park is hilly and highly undulating, with dominated tree
species such as Tectona grandis, Shorea robusta,
Buchanania latifolia, Terminalia arjuna, Emblica
officinalis, Madhuca indica and Rauwolfia serpentina.
The faunal diversity comprises of major carnivore such
as Tiger (Panthera tigris), Leopard (Panthera pardus),
Dhole (Cuon alpines) and other small carnivores
including Jungle cat (Felis chaus), Palm civet
(Paradoxurus hermaphroditus) as well as ungulates such
as Spotted deer (Axis axis), Sambar (Cervus unicolor),
Wild boar (Sus scrofa), Barking deer (Muntiacus
muntjak), Rhesus macaque (Macaca mulatta) and
Common langur (Semnopithecus entellus). In satpura
birds of prey like crested hawk eagle, black eagle and
crested serpent eagle were major predators of Ratufa
indica centralis (Datta, 1999; Kumbhar et al., 2012).
Also Mehta (1997) reported leopard attempted to prey on
giant squirrel.
Sampling
Line transect methodology was adopted
(Buckland et al., 2001; Jathanna et al., 2008) and
distance sampling methodology was used to estimate
population density of giant squirrel in our study area.
Field sampling was carried out in the months of
December to February 2011 2012. Dur-ing this period
39 permanent transects were established in different
habitat types including riparian patches. Each transect
was surveyed thrice by well trained observer be-tween
Gurjar et al., 2013
1087 Journal of Research in Biology (2013) 3(7): 1086-1092
06000900 hr. Each transects differed in length, the
average transect length was 2km to 4km. Every time the
species was detected group size, sighting distance and
angle of sighting were recorded. Sighting distances were
measured using lesser rangefinder and the angle of
sighting was recorded using a liquid filled compass. The
field protocols were followed described in Jhala et al.,
(2009). The density of Indian giant squirrel (IGS) was
calculated using DISTANCE program version 6.0 (Laake
et al., 1994). The best model was selected on the basis of
the lowest Akaike Information Criteria (AIC) (Burnham
et al., 1980; Buckland et al., 1993).

RESULTS AND DISCUSSION
A total of 35 Giant squirrel sights comprising
42 individuals were recorded during the study period in
total efforts of 276km. Analysis were done by fitting
different detection functions to the observed data for the
estimation of density. Based on minimum AIC value
(94.9), half normal with cosine proved to be the best fit
for giant squirrel data. As giant squirrel is a arboreal
species its visibility is very high when we compare it
with other terrestrial animals so detection in uniform
manner is normal, AIC value also supports the model
selection. The encounter rate was 0.12 0.06/km
walked, IGS known to be a solitary animal, maximum
two individuals were recorded in a group and mean
group size was calculated as 1.2 0.6 in Satpura Tiger
Reserve.
Studies conducted elsewhere on Indian Giant
Squirrel (IGS) have shown different estimates of
population density (Table. 2). The variation in different
Gurjar et al., 2013
Journal of Research in Biology (2013) 3(7): 1086-1092 1088
Figure 1: Location of Satpura Tiger Reserve in India.

estimates in different studies could be due to the different
habitat types in the different study areas; also seasonal
annual variation and observer differences put limits of
comparison. The present study is the first attempt to
provide baseline information on ecological density status
of Indian giant squirrel in Central Indian landscape
(Table. 1). IGS distribution in STR was observed in
Terminalia arjuna, Madhuca longifolia and Tectona
grandis. These trees are mostly used for feeding and
nesting (Kumbhar et al., 2012). Maximum IGS sightings
were recorded in riparian patches of churna, moist and
dry deciduous forest of watch tower and semi-evergreen
forest of Nimghan to pachmarhi. A viable population is
one that maintains its genetic vigor and potential for
evolutionary adaptation (Kumar et al., 2007), therefore
continuous monitoring of the population status of this
lesser-known mammal in central India should be given
high conservation priority. Excessive amount of
Gurjar et al., 2013
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Figure 2: Result of model fitted in the DISTANCE to estimate detection probability and effective
strip width of giant squirrel in Satpura Tiger Reserve.
Perpendicular distance in meters
Note: DS- estimate average group size; E(S) estimate expected value of cluster size; D estimate of density of
animal; N estimate no. of animals in specified area; Chi-square value P 0.969.
Table 1: Population density and average group size of Indian Giant Squirrel
(density /Km
2
) estimated in Satpura Tiger Reserve.
Parameter Point Estimate Standard Error Percentage Coefficient
of variation
95% Confidence Interval
DS 4.786 0.66 13.83 3.62 6.31
E(S) 1.169 0.59 5.05 1.05 1.29
D 5.595 0.82 14.73 4.17 7.49
N 6.000 0.88 14.73 4.00 7.00
poaching pressure and habitat fragmentation has been
reported in Orissa (Pradhan et al., 2012) which can leads
to population decline. We hope this baseline study will
encourage long-term study, which includes on nesting
breeding habits and resource availability of IGS
populations in Central Indian Forest. Further research
study about population status for this species and
conservation strategies in the central Indian landscape
are recommended.

CONCLUSION:
The present population density of Indian giant
squirrel 5.5 0.8 / Sq Km in Satpura tiger reserve in
central Indian forest is very important as it is first density
estimates from any central Indian forest and will provide
baseline data for future study. Present study is address
the issue of urgent need of survey the status, distribution
and abundance of Indian giant squirrel in central Indian
landscape.

ACKNOWLEDGE:
We are really grateful to Ravi Singh, Secretary
General and CEO, WWF-India and Principal Chief
Conservator of Forest (Wildlife), Chief Wildlife Warden,
Madhya Pradesh for give permission to conduct
phase-IV monitoring of predators and their prey in
Satpura Tiger Reserve. We would like to acknowledge
frontline staff of Satpura tiger reserve, Ratnesh and
Kamal Thakur for their extensive help in field work.

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Gurjar et al., 2013
Table 2: Density of Indian Giant Squirrel (individual/Km
2
) from other part of India.
Study site Density of IGS /Sqkm Authors
Anamalai Hills 11.4 - 64 Umapathy and Kumar 2000
Kudremukh NP 0.25 Madhusudan and Karanth 2002
Bandipur TR 2.36
Jathanna et al., 2008
Nalkeri 4.55
Sunkadakatte 4.86
Muthodi 10.19
Lakkavalli 12.25
Madumalai TR
2.9 Baskaran et al., 2011
1.6 Ramesh et al.,2009
Kalakad-Mudanthurai TR 1.7 Ramesh et al., 2012
Kakachi 1.42 Ganesh and Davidar 1999
Bhimashankar W Sanctuary
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15.89 Mehta et al.,2012

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Gurjar et al., 2013
Article Citation:
Mathews Plamoottil and Nelson P. Abraham.
Puntius viridis (Cypriniformes, Cyprinidae), a new fish species from Kerala, India.
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Puntius viridis (Cypriniformes, Cyprinidae),
a new fish species from Kerala, India
Keywords:
Fish, New species, Puntius parrah, Manimala River, Kallumkal.
ABSTRACT:

Taxonomic analysis of eight specimens of a cyprinid fish collected from
Manimala River, Kerala, India revealed that they present several morphological
differences from their congeners. The new species, Puntius viridis, is diagnosed by a
combination of the following characters: eyes clearly visible from below ventral side;
head depth lesser; one row of prominent elongated black spots on the middle of
dorsal fin; a black band formed of dark spots present outer to operculum. 25-26
lateral line scales; 4- 5 scales between lateral line and dorsal fin; moderate scales
on the breast region
1093-1104| JRB | 2013 | Vol 3 | No 7
An International
Authors:
Mathews Plamoottil and
Nelson P. Abraham.

Institution:
1. Government College,
Chavara, Kollam Dt, Kerala.
Pin code: 691583.

2. St.Thomas College,
Kozhencherry, Kerala.

Mathews Plamoottil.

Email Id:

Web Address:
Dates:
Received: 14 Aug 2013 Accepted: 02 Dec 2013 Published: 18 Jan 2014
Original Research
Print: 2231 6280;
Online: 2231- 6299.
ISSN No:
http://zoobank.org / urn:lsid:zoobank.org:pub:F091CFE1-4510-419E-89B4-EBE147BFD9D6
http://zoobank.org / urn:lsid:zoobank.org:act:7569C0D4-1236-473F-AE67-541C6A4C9A10

INTRODUCTION
The tropical Asian cyprinid genus Puntius
contains 120 valid species (Pethiyagoda et al., 2012).
The genus as currently known (Pethiyagoda et al., 2012)
is characterized by the absence of rostral barbels, last
unbranched dorsal fin ray smooth, dorsal fin with 3-4
unbranched and eight branched rays, anal fin with three
unbranched and five branched rays, lateral line complete
with 22- 28 pored scales, presence of free uroneural,
simple and acuminate gill rakers and presence of a post-
epiphysial fontanelle.
Jayaram (1991) revised the fishes of the genus
Puntius from the Indian region. He classified different
species of Puntius into 10 groups with 14 complexes.
But it is now understood that five lineages are present
within South Asian genus Puntius, which are recognized
as distinct genera namely Puntius, Systomus, Dawkinsia,
Haludaria and Pethia.( (Pethiyagoda et al., 2012;
Pethiyagoda, 2013); of these Puntius and Dawkinsia are
the common cyprinid fishes of the country. In Kerala
different species of Puntius preponderate in number than
any other scaled fresh water fishes.
Since the presently described specimens from
Manimala River did not have rostral barbels, possession
of smooth last unbranched dorsal ray and was similar in
morphology to the genus Puntius (sensu stricto), the
authors compared the specimens with comparative
materials of the currently known species in that genus
and found that the new species differs in enough
characters to distinguish it from other similar fishes of
the genus. So it is described here as a new species
Puntius viridis. The descriptions are based on eight
specimens collected from main stream of Manimala
River at Kallumkal.

Fishes were collected using cast nets and
preserved in 10% formalin. Methods used are those of
Jayaram (2002) and measurements follow standard
practices. In the table values of holotype as percentages
are given first, then ranges (holotype + paratypes) as
percentages followed by their mean values. Body depth
and body width were measured both at dorsal-fin origin
and anus, vertically from dorsal-fin origin to belly, and
from anus to dorsum, respectively.
Abbreviations
ZSI/WGRC/IR-Identified Register, Zoological
Survey of India, Western Ghats Regional Centre,
Kozhikode; ZSI/SRC-Zoological Survey of India,
Southern Regional Centre, Chennai; ZSI- Zoological
Survey of India, Kolkata; UOK/AQB- University of
Kerala, Department of Aquatic Biology, Kariavattom,
Thiruvananthapuram; CRG-SAC- Conservation
Research Group, St. Alberts College, Kochi; STC/DOZ-
St. Thomas College, Kozhencherry, Department of
Zoology; BDD- Body Depth at Dorsal origin; BDA-
Body Depth at Anal origin; BWD- Body Width at Dorsal
origin; BWA- Body Width at Anal origin; PROD-Pre
Occipital Distance; D-OD- Distance from Occiput to
Dorsal fin origin; LCP- Length of Caudal Peduncle; DCP
- Depth of Caudal Peduncle; DP-PL- Distance from
Pectoral fin to Pelvic fin; DPL-A- Distance from Pelvic
fin to Anal fin; DA-C- Distance from Anal fin to Caudal
fin; DAV- Distance from Anal to Vent; DVV- Distance
from Ventral to Vent; LMB- Length of Maxillary
Barbels; LLS- Lateral Line Scales; PDS- Pre Dorsal
Scales; PRPLS- Pre Pelvic Scales; PRAS- Pre Anal
Scales; CPS- Circum Peduncular Scales; LL/D- Scales
Between Lateral Line and Dorsal fin; LL/V- Scales
between Lateral Line and Ventral fin; LL/A- Scales
between Lateral Line and Anal fin; L/TR- Lateral
Transverse Scales; D- Dorsal fin; P- Pectoral fin; V-
Ventral fin; A- Anal fin; C- Caudal fin; HT- Holotype;
PT- Paratype.
Puntius viridis, sp. nov.,
http://zoobank.org / urn:lsid:zoobank.org:act:7569C0D4-
1236-473F-AE67-541C6A4C9A10
(Figures 1-4, 5. F & Tables 1 & 2)
Plamoottil and Abraham, 2013
Type materials examined
Holotype
ZSI/ WGRC/IR/2382, 81 mm SL, Kallumkal,
Manimala River, Kerala, India, 9200N, 76300E,
collected by Mathews Plamoottil, 21.08.2011.
Paratypes
ZSI FF 4932, 2 examples, 63- 74 mm SL,
Manimala River at Kallumkal, Kerala, India, collected
by Mathews Plamoottil, 10. 10. 2012.
ZSI/ WGRC/ IR/2383, 5 examples, 72- 76 mm SL,
Kallumkal, Manimala River, Kerala, India, coll.
Mathews Plamoottil, 21.08.2011.

Diagnosis:
Puntius viridis can be differentiated from
P. dorsalis in having a terminal mouth (vs. sub terminal
mouth), a comparatively short snout (22.7- 31.8 vs. 31.8
- 37.1 in % of HL), LL/V 3 (vs. 2) and caudal fin
with 18- 19 rays (vs.17). The new species differs from
Puntius sophore in having 10- 12 pre anal scales (vs. 13
pre anal scales in P. sophore), 3 scales between lateral
line and anal fin (vs. 4), a black band present outer to
operculum (vs. black band absent), a black blotch present
in front of occiput (vs. black blotch absent) and absence
of spot on the base of dorsal fin (vs. black spot present at
the base of dorsal fin), body depth at dorsal origin 31.5-
Figure 1: Puntius viridis, sp. nov, (fresh specimen), Paratype, 76 mm SL, ZSI/WGRC/IR/2383.
Figure 2: Puntius viridis, sp. nov, (preserved in formalin), Holotype, 81 mm SL, ZSI/ WGRC/IR/2382.

33.8 in % of SL (vs. 36.2- 37.3), eye diameter 26.1- 31.6
in % of HL (vs. 34.7- 36.0) and head depth 68.2- 80.0 in
% of HL (vs. 80.3- 86.7). The new species differs from
Puntius parrah in having nine pre dorsal scales (vs. 8 in
P. parrah), a deep black caudal spot (vs. diffused caudal
spot), green dorsal and caudal fin (vs. dusky dorsal and
caudal fin), longer head, 26.4- 31.1 % of SL (vs. 25.6-
26.0), shorter caudal peduncle, 16.3- 17.8 % of SL (vs.
19.1- 21.2) and shorter head depth (68.2-80.0 vs. 84.2-
89.5 % of HL); the new species differs from Puntius
madhusoodani in having 4- 5 scales between lateral
line and dorsal fin (vs. 4 scales), 8 branched rays in
dorsal fin (vs. 7), 5 branched rays in anal fin (vs. 6), a
deep black caudal spot (vs. diffused caudal spot) and
lesser body depth at dorsal fin origin (31.5- 33.8 vs. 34.5
- 36.2); the new species can be differentiated from
Puntius chola in having 8 anal fin rays (vs. 7 in P.
chola), 10-12 pre anal scales (vs. 12-13), 9- 10
circumpeduncular scales (vs. 11- 12), protrusible mouth
(vs. non- protrusible mouth) and a row of black spots
present in the middle of dorsal fin (vs. absent).
Description:
General body shape and appearance is shown in
Figures 1- 4. Morphometric data as in Table 1 and
meristic counts as in Table 2. Body laterally
compressed; dorsal and ventral profiles convex; region
from dorsal front to occiput a little bent, after sinking
down very slightly goes straight to snout tip; post dorsal
region slightly concave. Eyes situated considerably
behind and above the angle of jaws, protruding above the
surface of head and distinctly visible from below the
ventral side; inter orbital region slightly convex; nostrils
situated nearer to eyes than to snout tip and covered by a
flap originating from the anterior end; jaws equal, upper
jaw broader than lower jaw; tip of upper jaw a little
bulging and so can be easily demarcated from the rest of
it; barbels one pair maxillaries only, shorter than orbit,
feeble and never reach the eyes or nostrils; mouth
terminal, slightly upturned and protruding; width of gape
of mouth shorter than inter narial distance; operculum
rigid and moderately hard.
Dorsal fin originates considerably behind the
pectoral tip and a little behind the ventral origin, upper
margin fairly concave, first ray very minute, soft and
seemingly absent, commonly fused to second ray which
is slightly osseous, soft, tip a little filamentous, form a
little less than and above 1/3 of the third ray; third ray
osseous but not much strong, tip filamentous, inner
margin slightly roughened but not serrated. Last dorsal
ray branched to root and so considered as one. Pectoral
tip just reaches or reach nearer to ventral origin; its upper
margin convex. Ventral originates just in front of dorsal
origin and a little behind pectoral tip; its tip never
reaching anal origin, but only reaching the vent; upper
Figure 3: Dorsal fin of Puntius viridis Figure 4: Head region of Puntius viridis
margin of ventral fin convex; two scales present on either
side of base of ventral, one above the other, of this the
upper one soft and delicate, lower one more fleshy, form
2 of the length of ventral. Anal roughly rectangular,
upper margin fairly concave, originates a little in front of
dorsal tip, considerably behind the ventral tip and a little
behind anal opening; its tip never reach caudal base; no
prominent ridge on the base of anal; considerable
distance in between anal fin origin and vent; first anal
ray small; unbranched rays are slightly osseous; last anal
ray not divided to root. Caudal lobes equal.
Scales relatively large, not easily deciduous and
clearly countable; scales on the breast region moderate.
Lateral line passes through lower half of the body and
fairly distinct throughout.

Coloration:
Fresh specimens:
Dorsal and dorso lateral sides green to silvery
green; ventro lateral sides silvery green; eyes greenish
blue; a prominent yellowish green rectangular spot on
opercle; a black band formed of dark spots present outer
to operculum; a black blotch present just in front of
occiput, in the middle of it present a small elongated
depression; dorsal and caudal fins light green, pectoral
and anal light green to hyaline, distal end of anal black;
ventral hyaline to white. A row of distinct black spots
present on the middle of dorsal fin; a deep black caudal
blotch present well behind anal tip on 20-22 or 21-23 or
23-25 scales; 2- 3 rows of mid lateral scales have dark
spots at its base, so appear to have 2-3 broken lines on
mid lateral side.

Figure 5: General body shape and appearance of Puntius viridis and relative species.
Puntius dorsalis ZSI/F 2730 (coll. Francis Day) B. P. parrah ZSI/F 2718 (coll. Francis Day) C.P.
chola ZSI/F 2804 D. P. madhusoodani Paratype CRG-SAC 457 E. Puntius sophore ZSI/F 13827 F.
P. viridis Holotype, SL, ZSI/ WGRC/IR/2382.


SL.N0.

Characters
Puntius viridis sp. ov.
Mean

SD

P. parrah
ZSI/F2718,4934
(n=5)

P. madhusoodani
CRG/SAC 456- 459
(n=4)
HT Range HT+PT
(n=8)
1 Total length (mm) 103.0 91.2 -103.0 96.5 4.04 86.5 - 102.0 90.5 - 118.3
2 Standard Length (mm) 81.0 72.0 - 81.0 74.9 3.26 65.5 - 78.0 67.6 - 91.4
% SL
3 Head length 28.4 26.4 - 31.1 28.7 1.75 25.6 - 26.0 27.5 - 29.5
4 Head depth 22.2 19.7 - 22.9 21.6 1.13 21.6 - 24.0 20.7- 23.1
5 Head width 16.7 15.8 - 17.8 17.1 0.45 15.4 - 17.6 15.0 - 16.7
6 BDD 33.3 31.5 - 33.8 32.9 0.94 32.1 - 33.1 34.5 - 36.2
7 BDA 22.2 21.1 - 23.9 22.6 0.98 23.7 - 24.4 22.1 - 23.7
8 BWD 18.5 16.2 - 19.1 17.7 1.16 17.3 - 19.7 17.6 - 19.1
9 BWA 12.3 10.8 - 13.2 12.2 0.88 13.4 - 15.2 11.7 - 14.5
10 PROD 19.1 18.9 - 23.0 20.9 1.22 20.5 - 24.3 18.9 - 22.9
11 D-OD 30.6 30.4 - 31.7 30.9 0.31 24.3 - 29.8 29.0 - 32.9
12 Pre-dorsal length 50.6 48.2 - 54.8 52.2 1.61 50.0 - 52.1 49.3 - 50.6
13 Post-dorsal length 50.6 48.2 - 54.8 52.2 1.61 48.7 - 53.5 50.2 - 58.6
14 Pre-pectoral length 27.2 25.8 - 29.7 28.3 0.92 27.0 - 28.2 26.2 - 28.9
15 Pre-pelvic length 49.4 47.9 - 50.0 49.0 0.73 47.2 - 51.3 46.5 - 50.3
16 Pre-anal length 72.2 72.2 - 76.6 73.3 1.68 70.3 - 74.4 67.6 - 74.3
17 Length of dorsal fin 23.5 22.4 - 26.5 24.2 1.58 22.1 - 24.4 25.2 - 28.7
18 Length of pectoral fin 17.3 16.7 - 19.7 18.5 1.19 17.6 - 19.8 17.7 - 19.1
19 Length of pelvic fin 17.3 17.3 - 20.3 19.0 1.13 20.3 - 21.4 20.7 - 21.1
20 Length of anal fin 14.8 14.8 - 18.9 17.4 1.58 13.3 - 16.8 19.2 - 21.5
21 Length of caudal fin 29.5 29.3 - 30.0 29.6 0.20 28.4 - 32.1 24.8 - 27.0
22 Length of base of dorsal fin 18.5 17.6 - 19.2 18.5 0.60 18.0 - 21.0 19.0 - 20.0
23 Length of base of anal fin 9.8 9.8 - 11.1 10.7 0.43 12.0 - 15.4 9.0 - 12.0
24 Length of base of pectoral fin 4.3 4.1 - 5.3 4.5 0.48 3.3 - 4.2 3.7 - 4.1
25 Length of base of pelvic fin 5.2 5.0 - 6.9 5.9 0.77 4.2 - 5.4 6.0 - 7.1
Table 1: Morphometric characters of Puntius viridis and its relative species from Kerala
Preserved specimens:
Dorsal and upper lateral sides blackish green,
lower lateral and ventral sides whitish yellow; spot on
the operculum becomes brownish black colored; a
greenish line present above the ventral origin to caudal
spot which is distinct in some specimens in preserved
condition; pectoral, pelvic and anal becomes hyaline,
dorsal and caudal become dirty black, base of caudal
turns to black.
Distribution:
Puntius viridis sp. nov is presently known only
from Manimala River, Kerala, India.
Habitat:
Manimala River at Kallumkal the type locality of
P. viridis is blanketed by mud dominant sediments. Sand
occurs as discrete patches within the mud dominant
deposits. The depth and width of the channel at
Kallumkal ranges from 1 to 10 and 30 to 85 m
respectively. The reach has a bank height of 1 to 2 m
from the general water level. Riparian vegetation is
moderate. Dominant flora include Bambusa bambos,
B. vulgaris, Hibiscus tiliaceus and Ochreinauclea
missionis. The other species include Thespesia populnea,
Artocarpus heterophyllus, Areca catechu, Anacardium
occidentale, Aporosa lindleyana and Ficus exasperata.
Cynodon dactylon and Cymbopogon flexuosus are major
grass species in this area. Rasbora daniconius,
Osteobrama bakeri, Amblypharyngodon microlepis,
Dawkinsia filamentosa, Haludaria fasciatus, Puntius
parrah, Systomus subnasutus, Pethia ticto,
Gonoproktopterus kurali, Catla catla, Labeo rohita,
Labeo dussumieri, Cirrhinus mrigala, C. cirrhosus,
26 Length of base of caudal 13.6 13.5 - 14.2 13.8 0.34 12.2 - 14.1 12.4 - 13.8
27 Length of caudal peduncle 17.3 16.3 - 17.8 17.0 0.62 19.1 - 21.2 12.6 - 17.5
28 Depth of caudal peduncle 13.6 13.5 - 14.5 13.8 0.37 12.9 - 13.5 12.8 - 14.6
29 LCP/DCP 78.6 77.0 - 88.0 81.2 3.20 63.6 - 74.3 73.1 - 84.6
30 Width of caudal peduncle 7.4 5.5 - 7.4 6.5 0.77 4.1 - 5.4 6.2 - 6.6
31 DP- PL 21.0 21.0 - 21.6 21.4 0.20 20.4 - 20.9 22.8 - 25.0
32 DPL-A 24.2 23.8 - 25.0 24.3 0.60 24.3 - 26.8 25.0 - 28.9
33 DA-C 26.0 25.9 - 27.5 26.6 0.51 27.7 - 29.6 25.5 - 27.0
34 DAV 3.7 2.6 - 4.1 3.2 0.61 _ 4.8 - 6.6
35 DVV 22.8 19.1 - 22.8 21.2 1.29 23.0 - 25.6 22.4 - 23.4
% HL
36 Head depth 78.3 68.2 - 80.0 74.3 4.26 84.2 - 89.5 95.0 - 100.0
37 Head width 58.7 56.5 - 63.2 59.8 2.52 60.0 - 68.4 55.0 - 61.9
38 Eye diameter 30.4 26.1 - 31.6 29.6 2.07 32.5 - 36.8 27.5 - 33.3
39 Inter orbital width 39.1 31.6 - 40.9 37.5 3.37 42.1 - 42.5 37.5 - 41.9
40 Inter narial width 28.3 23.9 - 28.9 26.8 1.95 23.5 - 30.0 25.0 - 28.6
41 Snout length 30.4 22.7 - 31.8 29.1 3.39 26.3 - 30.0 28.6 - 30.0
42
Width of gape of mouth

26.1 23.0 - 27.3 25.5 1.53 28.9 - 30.0 25.0 - 27.6
43 LMB 17.4 13.0 - 21.1 17.8 3.69 15.0 - 17.6 14.3 - 15.0
Horabagrus brachysoma, H. melanosoma Mystus
indicus, Wallago attu etc are some of the co- occurring
species.
Etymology:
Species name comes from the Latin word viridis
meaning green, an adjective, given here in reference to
greenish colored body and fins of the new species.
Comparisons:
Puntius viridis is related to Puntius parrah,
P. madhusoodani, P. dorsalis, P. chola and P. sophore
(Figure 5). Puntius dorsalis (Jerdon, 1849) [Figure.5 A]
was described from the fresh water bodies of Madras
(Jayaram, 1991; Talwar & Jhingran, 1991; Pethiyagoda
et al., 2008). It differs from the new species in many
meristic and morphometric characters (Table 2). In
Puntius dorsalis a black spot present at the posterior
portion of the base of dorsal fin (vs. no black spot in the
present species), mouth sub terminal (vs. mouth
terminal), dorsal fin inserted nearer to caudal fin base
than tip of snout (vs. dorsal fin inserted in the middle
between snout tip and caudal base), 2 scales present in
between lateral line and pelvic fin (vs. 3 scales ),
caudal fin with 17 rays (vs. 18 or 19 caudal rays) and
snout length 31.8-37.1 (vs. 22.7- 31.8) in percent of head
length, dorsal fin inserted in front of ventral (vs. dorsal
originates a little behind ventral fin) and black spots
absent in the middle of dorsal fin (vs. one row of
prominent elongated black spots present on the middle of
dorsal fin).
Puntius parrah Day (1865, 1878 and 1889)
[Figure. 5. B] of Karavannoor River of Kerala shows
distinct differences to the new species. In P. parrah, a
dark bluish line present along mid lateral line, which is
more distinct in preserved state (vs. dark bluish line
SL
No
Counts

Puntius viridis (n=8) P.parrah ZSI/
F2718, STC/
DOZ 20 (n=5)
P. madhusoodani
CRG/SAC 456- 459
STC/DOZ 21(n=6)
P. chola
ZSI/F2203,
4009(n=2)
P. dorsalis ZSI/
F2730,ZSI/
SRC4954 (n=3)
P. sophore ZSI/
F13827, STC/
DOZ 22 (n=3)
Holotype

Range

Scale Counts
1
LLS

25 25 - 26 25 25 - 26 26 - 28 25 - 26 25
2
PDS

9 9 8 9 9 9 9
3
PRPLS

5 5 6 6 5 - 6 5 - 6 5
4
PRAS

11 10 - 12 14 14 12 - 13 11 - 13 13
5
CPS

10 9 - 10 10 10 11 - 12 9 - 10 10
6
LL/D

4 4 - 5 5 4 4 - 5 4 - 5 5
7
LL/V

3 3 3 3 3 - 3 2 3
8
LL/A

3 3 3 3 3 3 4
9
L/TR

5 / 3 5 -5 /3 5/4 5 / 3 5 / 4 5 / 2 5 / 4
Fin Ray Counts
10
D

iii , 8 iii , 8 iii , 8 iii , 7 iii , 8 iii , 8 iii , 8
11
P

i , 14 i , 14 i , 14 i , 14 i , 13-16 i , 14-15 i , 13-14
12
V

i , 8 i , 8 i , 8 ii , 8 i , 8 i , 7 i , 8
13
A

iii , 5 iii , 5 ii , 5 ii , 6 iii , 5 iii , 5 iii , 5
14
C

18 18 - 19 19 19 19 17 18
Table 2: Meristic Counts of Puntius viridis sp.nov and its relative species
absent in fresh or preserved condition in the new
species), eyes golden (vs. greenish blue), pectoral,
ventral and anal tinged with yellow (vs. pectoral and anal
light green to hyaline, ventral hyaline to white), dorsal
and caudal are dusky (vs. dorsal and caudal are green), 8
pre dorsal scales (vs. 9), 6 pre pelvic scales (vs. 5), 14
pre anal scales (vs. 10-12), dorsal fin originate just over
ventral fin (vs. dorsal fin originate a little behind ventral
origin), caudal spot diffused (vs. caudal spot deep black),
smaller head (25.6- 26.0 % of SL vs. 26.4- 31.1 % of
SL), greater head depth at occiput, 84.2- 89.5 % of HL
(vs. 68.2- 80.0 % of HL), longer anal fin base (12.0- 15.4
% of SL vs. 8.8- 11.1), longer caudal peduncle (19.1-
21.2 % of SL vs. 16.3- 17.8) and greater distance
between ventral to vent (23.0- 25.6 % of SL vs. 19.1-
22.8). Above all, in the present species, just in front of
occiput a black blotch present, in the middle of which is
a small elongated depression, a black band present outer
to operculum, 2-3 broken lines on mid lateral side, a row
of elongated green dots on dorsal fin and a row of
distinct black spots present in the middle of the anal
which are all absent in P. parrah.
Puntius viridis sp. nov resembles Puntius chola
(Hamilton) [Figure. 5. C] of Gangetic plains in having a
blotch on caudal base, possession of a single pair of
maxillary barbels and in the number of ventral fin rays
(Hamilton, 1822; McClelland, 1839; Nath & Dey, 2000);
however, the new species shows differences to P. chola
in a number of characters. In P. chola anal fin has seven
rays (vs. eight rays in new species), no scale like
appendants above ventral fins (vs. an axillary ventral
scale present), a slight ridge present along the middle of
lower jaw (vs. no ridge along the middle of lower jaw),
arch of the back rising abruptly from the nape to the base
of the dorsal (vs. arch of back rising gradually from the
nape to the base of dorsal), a dark mark present along the
base of anterior dorsal ray (vs. dark mark absent ), lateral
line scales are 26- 28 (vs. 25- 26), pre anal scales 12- 13
(vs. 10-12), circum peduncular scales 11- 12 (vs. 9-10),
width of gape of mouth 19.0- 23.0 (vs. 23.0- 27.3), eyes
not visible from below the ventral side (vs. eyes
protruding above the surface of head and distinctly seen
from below ventral side), mouth not protrusible (vs.
mouth fairly protruding), no black band present outer to
operculum (vs. a black band present outer to operculum),
no black blotch in front of occiput (vs. a black blotch
present in front of occiput) and no black spots present in
the middle of dorsal fin (vs. a row of distinct black spots
present in the middle of dorsal fin).
The new species can also be easily distinguished
from Puntius madhusoodani [Figure.5. D] described by
Kumar et al., (2011) from Manimala River. In
P. madhusoodani, 4 scales present between dorsal fin
and lateral line (vs. 4- 5 scales in the new species),
dorsal side dusky black (vs. dorsal side greenish), dorsal
fin with seven branched rays (vs. dorsal fin with eight
branched rays), ventral fin with two unbranched and
eight branched rays (vs. ventral fin with one unbranched
and eight branched rays), anal with two unbranched and
six branched rays (vs. anal fin with three unbranched and
five branched rays), branched rays of dorsal and anal
rays black (vs. branched rays of dorsal and anal not
black), absence of spots except at caudal base (vs.
presence of spots other than on caudal base such as a
black blotch just in front of occiput, a thin dark band
present outer to operculum and a row of green dots
present in the middle of dorsal fin), mouth sub terminal
(vs. mouth terminal), pelvic fin slightly posterior to
dorsal origin (vs. pelvic origin just in front of dorsal
origin), body depth at dorsal origin 34.5- 36.2 (vs. 31.5-
33.8) and length of anal 19.2- 21.5 (vs. 14.8- 18.9) in
percent of standard length.
Puntius sophore (Hamilton), [Figure. 5. E]
described from Gangetic provinces shows many
similarities to present species in meristic and
morphometric features (Misra, 1962; Rema devi, 1992;
Datta & Srivastava, 1988; Talwar and Jhingran, 1991;
Jayaram, 2010). In P. sophore, a black spot present at
the root of the dorsal fin (vs. black spot absent at the root
of dorsal fin in the new species), barbels absent (vs. one
pair of maxillaries present), a faint band present on the
lateral side (vs. lateral band absent), no black band
present outer to operculum (vs. a black band present
outer to operculum), no black blotch in front of occiput
(vs. a black blotch present in front of occiput , in the
middle of which a small elongated depression), no black
spots present in the middle of dorsal fin (vs. a row of
distinct black spots present in the middle of dorsal fin),
body depth at dorsal origin 36.2- 37.3 (vs. 31.5- 33.8),
pre anal length 71.2- 72.2 (vs. 72.3- 76.6), length of
pelvic fin 20.7- 22.0 (vs. 17.3- 20.3) and distance from
pelvic to anal fin 25.8- 27.6 (vs. 23.8- 25.0) all in percent
of SL; head depth at occiput 80.3- 86.7 in % of HL (vs.
68.2- 80.0) and eye diameter 34.7- 36.0 in % of HL (vs.
26.1- 31.6).

CONCLUSION
Puntius viridis is a barb usually caught along
with Puntius mahecola and Dawkinsia filamentosa. It is
an edible fish can usually be collected by small- meshed
gill nets. They show similarities with Puntius parrah
and P. madhusoodani of Kerala, P. dorsalis of Madras
and Puntius chola of northern parts of India. They can
be easily identified from their congeners in having a
black band formed of dark spots present outer to
operculum and a row of distinct black spots present on
the middle of dorsal fin. They have also a less deep
head. It is expected that further research works may
unveil its more biological aspects.
Comparative material
Puntius dorsalis: 27.10.95, 1 example, 62 mm
SL, Thunakadavu dam, Parambikulam wild life
sanctuary, Kerala, ZSI/WGRC/IR 8466, coll. P.M.
Sureshan, identified by K. C. Gopi; 23.2.2000, 2
examples, 56- 63 mm SL, Pampa River at Parumala,
Kerala, ZSI/WGRC/IR/10379, coll. K. C. Gopi; 11.02.
58; 1 example, 53 mm SL, Usteri tank, 7 miles north
west of Pondicherry, ZSI/F 2801, coll. A.G.K. Menon;
16.02. 1996, 2 examples, 52- 53 mm SL, Sethumadai
canal, Indira Gandhi Wild Life sanctuary, Tamil nadu,
ZSI/SRC/F 4954, coll. M.B. Reghunathan; undated, 1
example, Madras, ZSI/F 2730, coll. Francis Day;
undated, 1 example, 53 mm SL, Tunga River at
Shimoga, ZSI/F 12320/1, coll. H.S. Rao; undated, 5
examples, 55- 62 mm SL, Cauvery River, Coorg,
Karnataka, ZSI/F 12319/1, coll. C.R. Narayan Rao;
Puntius parrah: 10.01. 2012, 4 examples, 65.5-
78.0 mm SL, Arattupuzha, Karavannoor River,
Iringalakuda, Kerala, ZSI FF 4934, coll. Mathews
Plamoottil; 15.12.1994; 1 example, 60 mm SL, Kuruva
Island, Wayanad, ZSI/WGRC/IR/742, coll. C.
Radhakrishnan; 24.03.1997, 1 example, 44 mm SL,
Parambikulam WLS, ZSI/WGRC/IR/10696, coll. K. C.
Gopi; 10.8.2001, 2 examples, 100.0- 103.0 mm SL,
Achankoil River, UOK/AQB/F/ 102, coll. Bijukumar;
undated, 1 example, Kariavannoor River, Kerala, ZSI/F
2718 Syntype, coll. Francis Day; 08.05. 1977, 6
examples, 71 mm- 94 mm SL, Cauvery River at
Chunchinagatte, ZSI/SRC Uncat, coll. K. C. Jayaram.
Puntius chola: 08.11.1939, 1 example, 41.5 mm
SL, Soni Gaon Bheel, Lokpa, Batipara, Assam, ZSI/F
2203, coll. S.L. Hora; 1963, 1 example, 54 mm SL,
Sukla Talai, Jhalwar, Rajasthan, ZSI/F 4009/2, coll. N.
Majumdar & R.N. Bhargava; 18.03.1958, 2 examples,
32.5- 55 mm SL, Raxanal, Bihar, ZSI/F/2804/2, coll.
Keval Singh; 3 examples, 50- 62 mm SL, Rajastan,
ZSI/F/4379/2, coll. Birla college, Pilani; 1 example, 71
mm SL, Mahanadi Irrigation Canal, Rudri, Orissa, ZSI/F
13082/1, coll. H.S. Rao.
Puntius madhusoodani: 17.11.2010, Holotype,
91.43mm SL, Manimala River, near Thirumoolapuram,
Thiruvalla, Kerala, CRG-SAC 456, coll. K.
Krishnakumar; 17. 11. 2010, 3 examples, 67.6 -
80.91mm SL, Manimala River, near Thirumoolapuram,
Thiruvalla, Pattanamthitta District, CRG-SAC 457 459
paratypes, coll. K. Krishnakumar and Benno Pereira.
Puntius sophore: 10.05.2012, 2 examples, 58- 59
mm SL, Serrampore, River Ganges, Kolkata, ZSI FF
4938, Coll. Mathews Plamoottil; 20.06. 1963, 4
examples, 62.5- 70.0 mm SL, Sukla Talai, Jhalawar,
Rajasthan, ZSI/F 4008/2, coll. N. Majumdar & R. N.
Bhargava; 24.10.1939, 1 example, 40 mm SL, Siwane
River, east of Hazaribagh Barthi Road, ZSI/F 13827,
H.S. Rao; 22.06.1963, 4 examples, 66- 102 mm SL,
Gadhuli Talai, Shergarh, Rajasthan, ZSI/F 4023, SE
Rajastan Survey of ZSI; 30.06.1983, 4 examples, 58.0-
67.5 mm SL, Talbi, N. of Bimmal Railway station, ZSI/F
4029/2, S. E. Rajasthan Survey of ZSI.

ACKNOWLEDGEMENTS
First author acknowledges the University Grants
Commission of India for sanctioning Faculty
Development Programme to undergo research. Both the
authors acknowledge the Principal, St. Thomas College,
Kozhencherry for providing the facilities.

REFERENCES
Datta MJS, Srivastava MP. 1998. Natural history of
fishes and systematics of fresh water fishes of India.
Narendra Publishing House, Delhi, 178-196.

Day F. 1865. The Fishes of Malabar. Bernard Quaritch,
London., 208-211.

Day F. 1878. The fishes of India: being a natural history
of the fishes known to inhabit the seas and fresh waters
of India, Burma, and Ceylon. William Dawson & Sons,
London, 556-574.

Day F. 1889. Fauna of British India including Ceylon
and Burma. Fishes. I, Taylor and Francis, London, 209-
334.

Hamilton F. 1822. An account of fishes found in the
River Ganges and its branches. Edinburgh Hurst,
Robinson & Co, London, 312-389.
Jayaram KC. 1991. Revision of the genus Puntius
Hamilton from the Indian region. Records of Zoological
Survey of India, Occasional Paper No. 135, 178.

Jayaram KC. 2002. Fundamentals of Fish Taxonomy.
Narendra Publishing House, Delhi. 53-65.

Jayaram KC. The Freshwater fishes of the Indian
region. Narendra Publishing House, Delhi.; 118-134.

Jerdon TC. 2010. On the freshwater fishes of southern
India. Madras Journal of Literature and Science, 15 (2):
302- 346.

Kumar KK, Pereira FGB and Radhakrishnan KV.
2011. Puntius madhusoodani (Teleosti: Cyprinidae), a
new species of barb from Manimala River, Kerala, South
India. Biosystematica, 5 (2); 31- 37.

McClelland J. Indian Cyprinidae. 1839. Cosmo
Publications, New Delhi, 246.

Misra KS. 1962. An aid to the identification of the
common commercial fishes of India & Pakistan.
Records of Indian Museum, 57(1-4): 320.

Nath P, Dey SC. 2000. Fish and fisheries of North
Eastern India (Arunachal Pradesh). Narendra Publishing
House, Delhi, 39-43.

Pethiyagoda R, Silva A, Maduwage K and
Meegaskumbura M. 2008. Puntius kelumi, a new
species of cyprinid fish from Sri Lanka (Teleostei:
Cyprinidae). Ichthyological Exploration of Freshwaters,
19 (3): 201- 214.

Pethiyagoda R, Meegaskumbura M and Maduwage
K. 2012. A synopsis of the South Asian fishes referred to
Puntius (Pisces: Cyprinidae). Ichthyological Exploration
of Freshwaters, 23 (1): 69-95.

Pethiyagoda R. 2013. Haludaria, a replacement
generic name for Dravidia (Teleostei: Cyprinidae).
Zootaxa, 3646 (2): 199.

Remadevi K. 1993. On a small collection of fish from
Javadi hills, North Arcot district, Tamil Nadu. Records
of Zoological Survey of India.; 91(3-4): 353-360.

Talwar PK, Jhingran A. 1991. Inland fishes of India
and adjacent countries. Oxford and IBH Publishing Co.
Pvt. Ltd, Delhi, 250-286.

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A new species of Agathoxylon Hartig from the Sriperumbudur
formation, Tamil Nadu, India
Keywords:
Agathoxylon, Sriperumbudur Formation, Upp. Jurassic-Low Cretaceous.
Abstract:

Sriperumbudur Formation is one of the Upper Gondwana rock Formations
found along the Palar basin, Tamil Nadu, India. The rock units found in this Formation
are arenaceous and argillaceous, consists of green shales, clays and sandstones with
limestone intercalations. These shales contain animal and plant remains of Upper
Jurassic-Lower Cretaceous age. The present work is about a piece of petrified
secondary wood of conifer having affinity with Araucariaceae. Based on the
anatomical characters the present wood is identified as a new species of Agathoxylon
Hartig.
1105-1110 | JRB | 2013 | Vol 3 | No 7

An International Scientific
Research Journal
Authors:
Kumarasamy D.

Institution:
Department of Botany,
Annamalai University,
Annamalainagar 608 002,
Tamil Nadu, India.

Kumarasamy D.

Email:

Web Address:
Dates:
Received: 14 Aug 2013 Accepted: 21 Sep 2013 Published: 18 Jan 2014
Article Citation:
Kumarasamy D.
A new species of Agathoxylon Hartig from the Sriperumbudur formation, Tamil Nadu, India.
Original Research

INTRODUCTION
The out crops of sedimentary rocks exposed in
patches all along the eastern shoreline of Indian
Peninsula starting from Cuttack in Orissa to Sivagangin
Tamil Nadu are collectively referred to as the East Coast
Gondwanas. These exposures occur along the Mahanadhi
basin, the Krishna-Godavari basin, the Palar basin and
the Cauvery basin. The upper Gondwana exposures
found along the Palar basin are divided into the lower
Sriperumbudur Formation and the upper Satyavedu
Formation. Equivalent to these two Formations there is a
marine Formation known as Avadi Formation
(Kumaraguru,1991).
The Upper Gondwana rocks exposed near
Sriperumbudur are part of a large Sriperumbudur
Formation found along the Palar basin (Kumarasamy and
Jeyasingh, 1995). The rock units found in this formation
are arenaceous and argillaceous, consist of green shales,
clays and sandstones with limestone intercalations.
These shales contain both marine animal and
plant remains of Upper Jurassic-Lower Cretaceous age.
These fossilferous shales are covered by the recent
lateritic and alluvial Formations.
Plant fossils found in this Formation includes
impressions of leaves of petridophytes and gymnosperms
and petrifield woods of gymnosperms. Many
publications came out regarding the fossils found in this
Formation, they are Feistmantel, 1879; Seward and
Sahni, 1920; Sahni, 1928 and 1931; Suryanarayana, 1953
and 1954; Ramanujam and Srisailam, 1974; Ramanujam
and Varma, 1977 and 1981; Varma, 1983 and 1984;
Varma and Ramanujam, 1984; Jeyasingh and
Kumarasamy, 1994a, 1994b and 1995; Kumarasamy and
Jeyasingh, 1995, 2004 and 2007. The present work is
about the observation of a new species of Agathoxylon,
from this Formation.

The present observation is about a piece of petrified
secondary wood (SPR/VK/52) collected from Vallakottai, a
village near Sriperumbudur (Formation named after this
town). The specimen was sectioned using rock cutting
and grinding machine. Thin sections (TS, TLS and RLS)
were prepared and observed under light microscope.
Photomicrographs were prepared using Olympus digital
camera attached with Olympus microscope.
Agathoxylon aptiana sp. nov.
Holotype : Specimen-SPR/VK/52
Slides : SPR/VK/52/1, 2, 3 and 4
Type locality : Vallakottai
Stratigraphic horizon : Sriperumbudur Formation, Upper
Jurassic-Early Cretaceous
Etymology : Named after the probable
age (Aptian) of the sediment
from where the specimen was
picked up.
Description (Fig. 1-a,b,c,d and e)
The study is based on a single piece of
decorticated pycnoxylic wood, measuring 5 cm long and
4 cm wide. The specimen is impregnated with ferrous
compounds. Growth rings distinct, almost straight,
almost equal, 600-710 mm (26-33 cells) wide. All
growth rings have more of early wood than late wood
(four rows of tracheids in average). Tracheids are
regularly arranged in radial rows. Transition from early
wood to late wood gradual. No reaction wood and false
ring. Early wood tracheids 2.0-3.3 mm long, radially
15-50 m (average 24.7 m) wide, rectangular to
circular. Radial wall pits mostly uniseriate, in some
places it is biseriate, alternate; pits bordered, circular,
contiguous, 12.5 m in size. Aperture elliptic, crossed,
6.25 m long and 2.5 m wide. Tracheids per mm
2

are 1599. Late wood tracheids 10.0-23.7 m (average
11.1 m) in radial diameter. Rays uniseriate, a few are
partially biseriate, 1-19 (average 6) cells high,
homocellular, cells 22.3 m long and 17.5 m wide.
Kumarasamy, 2013
Both tangential and horizontal walls are smooth. Radial
wall pits 3-9, circular, bordered, 7.5 m wide, tightly
packed. Aperture circular, ray cells spanning 2-3
tracheids, end walls vertical. Vertical parenchyma, resin
tracheids or resin canals are completely absent.
Diagnosis
Wood pycnoxylic, growth rings distinct. Only
radial wall of the tracheids are pitted. Radial wall pits
uni-biseriate, alternate, contiguous, circular with
elliptical crossed apertures, cross field pits 3-9, circular
and contiguous. Rays simple, uniseriate, 1-19 cells high;
xylem parenchyma and resin tracheids are absent.
The present wood shows alternate, uni-biseriate
pits (araucarioid pitting) on the radial wall of the
tracheids, uniseriate rays, and 3-9 pits per cross field.
These characters indicate that the present wood having
affinity with Araucariaceae.

DISCUSSION
There are sixteen morphogenera of fossil plants
have araucarian affinity. They are Agathoxylon Hartig,
Araucariopsis Caspary, Araucarioxylon Kraus in
Schimper, Araucarites Endlicher Sensu Goppert,
Baieroxylon Greguss, Cedroxylon Kraus in Schimper,
Kumarasamy, 2013
Fig. 1. Agathoxylon aptiana. a) transverse section showing growth ring, b) tangential longitudinal section
showing uniseriate rays, c) radial longitudinal section showing alternate pitting, d) tracheid radial wall
pits showing crossed apertures and e) gross field pits.
100m a
100m b
5m d 50m
c
5m e

Cordaioxylon Lignier, Cordaioxylon Lignier,
Cormaraucarioxylon Lignier, Dadoxylon Endlicher,
Dammaroxylon Schultze-Motel, Palaeoxylon Brongniart,
Peuce Lindley and Hutton, Pinites Witham,
Platyspiroxylon Greguss, Simplicioxylon Andreanzsky.
Among these names Araucarioxylon and Dadoxylon are
considered to be invalid names. Agathoxylon Hartig is
the earliest validly published name that can be used to
name fossil woods with an Araucarioxylon-type anatomy
(Philippe, 1993 and 2011)
So far, there are three species Araucarioxylon
r epor t ed fr om t hi s f or ma t i on namel y
A. rajivii (Jeyasingh and Kumarasamy (1994a)),
A. giftii (Jeyasingh and Kumarasamy (1994a)) and
A. mosurense (Jeyasingh and Kumarasamy (1995)). The
present fossil wood differ from A. rajivii in having
3-9 cross field pits, whereas in the latter wood there are
1-2 cross field pits per field, similarly in A. giftii the
cross field pits are 1-3. In A. mosurense the rays are
1-3 seriate, where as in the present wood the rays are
exclusively uniseriate.
The present specimen superficially resembles
Araucarioxylon bikanerense reported by Harsh and
Sharma (1988) from the tertiary deposits of Rajasthan
and A. agathioides reported by Krausel and Jain (1964)
from the Rajmahal hills. But the present specimen differs
from A. bikanerense in having uniseriate pits on the
radial walls of the tracheids, whereas in A. bikanerense
the radial wall pits upto triseriate. A. agathioides differs
from the present specimen is having frequent resin
tracheids but in the present specimen there are no resin
tracheids at all.
In the presence of biseriate radial wall pits with
elliptical, crossed apertures, 3-9 cross field pits per field
and the complete absence of xylem parenchyma and
resin tracheids, the present specimen stands apart from
all other species, so it is assigned to a new species.
So far, many species of fossil conifer woods
reported from this formation viz. Cupressinoxylon
coromandelianum Sahni (1931), M. thirumangalense
Suryanarayana (1953), Dadoxylon rajmahalense
Suryanarayana (1954), Araucarioxylon rajivii Jeyasingh
and Kumarasamy (1994a), A. giftii Jeyasingh and
Kumarasamy (1994a), A. mosurense Jeyasingh and
Kumarasamy (1995), Cupressinoxylon gondwanensis
Kumarasamy and Jeyasingh (2004) and Sahnioxylon
savitrii Kumarasamy and Jeyasingh (2007) have been
reported from this formation. Apart from these petrified
woods, many impression fossils of petridophytes and
gymnosperms were reported from this Formation
(Jeyasingh and Kumarasamy, 1994b; Kumarasamy and
Jeyasingh, 1995).
Recently a species of Agathoxylon was also
reported from this Formation (Kumarasamy, 2013). This
species (Agathoxylon gondwanensis) differs from the
present species in having one pit per cross field and long
xylem rays (1-39 cells high).
In general the overall climate during the
deposition of the sedimentary rocks in the Palar basin
should have been warm, humid and uniform. This is
indicated by the abundance of cycodophyte foliage in
these sediments. However, there must have been yearly,
seasonal variations as evident from the distinct growth
rings found in all the secondary wood pieces coming
from this formation. Most of the wood pieces show C
type growth-rings (as per Creber and Chaloner, 1984) in
which the early wood is more than the late wood and the
transition from the early wood to late wood is gradual.
These features indicate that the climate of this region was
almost uniform through the growing season except at its
close.

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Article Citation:
Dilworth LL, Brown KJ, Wright RJ, Oliver MS

and Asemota HN.
An assessment of bioactive compounds and antioxidants in some tropical legumes,
seeds, fruits and spices.
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An assessment of bioactive compounds and antioxidants in some tropical
legumes, seeds, fruits and spices
Keywords:
Antioxidants, bioactive compounds, spices, legumes, seeds
ABSTRACT:
Objective:
The main objective of this research was to assess bioactive compounds,
antioxidant potential and mineral concentration of commonly consumed foods as well
as underutilized ones for improved health and food security.
Methods:
Twelve food samples were assessed for minerals, flavonoids, IP
6
, total
polyphenols and antioxidant activity. IP
6
was determined by anion exchange
chromatography while flavonoids, polyophenols, minerals and antioxidant activity
were determined by standard methods.
Results:
The highest concentrations of IP
6
were recorded in legumes and corn while
appreciable levels were also found in golden apple and sorrel samples. The highest
concentrations of flavonoids and total polyphenols were found in non-
leguminaceaous samples. Pimento and ginger samples recorded highest antioxidant
activity (p<0.05) with values comparable to the standard ascorbic acid while pumpkin
seeds and onion samples recorded lowest antioxidant activities. Mineral
concentrations varied with the samples of pimento, golden apple and sorrel having
highest calcium concentrations. Sorrel, ginger and pimento recorded highest iron
concentrations, while zinc levels were as highest in both hulled and unhulled pumpkin
seed samples. Okra samples recorded the highest copper concentrations.
Conclusion:
Food samples analysed are rich in minerals, bioactive compounds and
antioxidants hence their increased exploitation for nutraceutical and nutritional
benefits are advocated. Data from this study argues well for increased production and
consumption of rarely consumed pumpkin and jackfruit seeds in light of their
nutritional profile and antioxidant activity. Most samples assessed are valuable in
supplementing nutrient-poor diets.
1182-1194 | JRB | 2014 | Vol 3 | No 7
An International
Authors:
Dilworth LL
*
, Brown KJ,
Wright RJ, Oliver MS

and
Asemota HN.

Institution:
Department of Basic
Medical Sciences,
University of the West
Indies, Mona campus.

Dilworth LL.

Email Id:

Web Address:
Dates:
Received: 31 Jan 2014 Accepted: 17 Feb 2014 Published: 28 Feb 2014
Original Research

INTRODUCTION
In light of concerns regarding food security and
quality, there is great interest in ascertaining the
nutritional benefits of foods commonly consumed
throughout the tropics. Functional food researchers
generally agree that in addition to macronutrients, it is
also important to assess minerals as well as levels of
bioactive compounds that may contribute to the overall
quality and health benefits of foods consumed by a wide
cross section of people in different geographical
locations. To that effect, assessing the antioxidant
activities of food samples is also important as it indicates
their ability to counteract the effects of free radicals. Free
radicals are independently existing atoms or molecules
that have one or more unpaired electrons (Williams
et al., 2006). They are generated daily in living systems
arising from the metabolic processes that form a part of
normal aerobic metabolism (Saha et al., 2008). The
increased incidences of many diseases including cell
tumours, type II diabetes mellitus and coronary heart
diseases are attributed to the effects of highly active free
radicals (Marinova et al., 2005; Olajire et al., 2011).
Throughout the Caribbean, there are many food
crops which are believed to possess therapeutic
properties. These beliefs are largely based on tradition
and have resulted in increased interest in the area of
ethnopharmacology. It is now theorized that traditional
medicine has immense value and the therapeutic
properties of foods may be due in part to the presence of
bioactive compounds (Sreeramulu et al., 2013). The
development of an industry from this knowledge is
considered an important contributor to economic growth
in the tropics (Dilworth et al., 2013). Some of the foods
of interest are spices and condiments including pimento,
ginger, onions, okra and sorrel that are reported to
possess important health benefits (Rubio et al., 2013;
Kaefer and Milner, 2008; Tsai et al., 2014; Prez-
Gregorio et al., 2014). Other foods of interest include
legumes and seeds including corn, pumpkin seeds,
jackfruit seeds, pigeon peas, broad beans, kidney beans
as well as golden apple (Hanson et al., 2014; Swami
et al., 2012; Kalogeropoulos et al., 2013; Islam et al.,
2013). Some of these foods are commonly consumed and
are reported to have a myriad of health benefits while
others are not commonly consumed but are easily
available and also have health promoting properties
which should be explored. The health benefits and
reported underutilization of some samples along with the
potential economic benefits of their incorporation into
mainstream consumption prompted research interest in
the food samples selected.
Since antioxidants are shown to significantly
delay or prevent the oxidation of easily oxidizable
substances, there is now an increased interest in the role
of natural antioxidants from different food sources.
Inositol hexakisphosphate or IP
6
(also known as phytic
acid or phytate when in salt form), is also thought to play
a role in antioxidant activity of cells. IP
6
is the principal
storage form of phosphorus in many plant tissues,
especially bran and seeds, where it exhibits antioxidant
properties via chelation of hydroxyl radicals (Graf and
Eaton, 1990; Johnson et al., 2000). IP
6
concentrations in
most of the food samples previously mentioned are not
known and therefore warrant investigation.
Minerals are an important contributor to the
nutritional value of foods as they play significant roles in
many essential metabolic processes. They are important
in cognitive development, function as enzyme cofactors,
and play important roles in structural and epithelial
integrity among numerous other functions. Reduced
levels and bioavailability of minerals is thought to be a
major health challenge in developing countries. There is
however, a paucity of information regarding overall
antioxidant properties and health benefits of many
commonly eaten foods. In light of the current boom in
the nutraceutical industry, it is important to assess their
antioxidant properties since this will positively
contribute to their marketability. This will also enhance
Dilworth et al., 2014
the commercial viability of the region since specific
foods and their value added products can be marketed for
economic development.
This study was geared at assessing the nutritional
value of foods delivered to the market for consumption
by the local population, since the average consumer
purchases food from the market and not directly from the
farm. Checks were done to ensure that all samples were
delivered to the market directly from the farm within
three days or less since older samples may have reduced
bioactive compounds and antioxidant activity owing to
improper storage. This research was aimed at
ascertaining antioxidant properties, bioactive compounds
and mineral concentration of commonly consumed foods
while assessing some other uncommon foods for
incorporation into mainstream consumption or for use as
nutraceuticals.

METHODOLOGY
MATERIALS:
Chemicals and Reagents were purchased from
Sigma-Aldrich Co. (MO, USA).
Samples
A wide variety of commonly eaten foods
including tuber crops, fruits, vegetables, condiments and
spices were selected for analyses. They were as follows:
Kidney bean (Phaseolus vulgaris), Butter bean
(Phaseolus lunatus), Pigeon peas (Cajanus cajan), Okra
(Hibiscus esculentus), golden apple (Spondias dulcis),
Jackfruit (Artocarpus heterophyllus), Sorrel (Hibiscus
sabdariffa), Onion (Allium cepa), Ginger (Zingiber
officinale), Pimento (Pimenta dioica) and Corn (Zea
mays). Samples were collected from the main market in
the city of Kingston, Jamaica, then taken to the
laboratory, washed and oven dried to a constant weight.
Samples were then crushed in a General electric motor
and industrial system laboratory mill with the mesh size
of 0.2 mm and stored frozen for further use.

METHODS
Determination of Minerals
The minerals calcium, copper, zinc and iron were
determined by standard methods (AOAC, 2005). A
specified amount of ground sample was completely
ashed followed by acid digestion and dilution with
deionized water. Samples were read using a Unicam 939
atomic absorption spectrophotometer equipped with
background correction and cathode lamps. Accuracy of
the analytical method was confirmed through a series of
certified analyses on reference materials. Appropriate
spikes were added to specific samples for recovery
determinations.
Total phenol
Total phenol levels were determined by a
modification of the Folin-Ciocalteu assay method as
described by Sun et al., (2006) and Prasad et al., (2010).
Following methanol extraction, 0.5 mL of Folin reagent
was added to samples and then Na
2
CO
3
was also added
.
Samples were vortexed and incubated, diluted with
deionized water, centrifuged and absorbance read at
725 nm. A standard curve for gallic acid was done based
on a similar procedure as outlined above. Extrapolations
for total polyphenol concentration were then carried out
from the curve and values given as mean SD mg gallic
acid equivalents/mL.
DPPH radical scavenging activity:
DPPH radical scavenging activity was
determined by slight modifications of methods outlined
by Matkowski et al., (2008), Veeru et al., (2009) and
Hasan et al., (2006). Plant extracts were double extracted
with methanol for 24 hours then rotor evaporated to
dryness and the DPPH assay was carried out to
determine the concentration of each extract required to
cause 50% inhibition. Samples were read at 517 nm
against a pure methanol blank in duplicates and
the percentage inhibition was determined according to
the equation below. IC
50
values were determined
from the graph of the percentage inhibition

against extract concentration.
Flavonoids
Total flavonoid content was assessed by the
aluminum chloride colorimetric assay as previously
reported (Marinova et al., 2005). An aliquot of the
methanolic extract was centrifugated and added to
deionized water, sodium nitrateand aluminium chloride
.
Sodium hydroxide was then added and the volume made
up to 10 mL with deionized water. Solutions were mixed
thoroughly and the absorbance was read at 510 nm
against a reagent blank. Total flavonoid content was
expressed as catechin equivalents (CE)/100 g dry mass.

IP
6

Assessment of IP
6
was done by a method
previously described by Siddhuraju and Becker (2001). It
involved a colorimetric method in addition to ion
exchange purification. Duplicate ground samples were
stirred with HCl at room temperature followed by
centrifugation. Aliquots were diluted with distilled water
and the pH was adjusted to 6. The diluted extract was
quantitatively transferred to a column with anioinic
exchange resin. Inorganic phosphate was eluted with 0.1
M NaCl while IP
6
was eluted with 1M NaCl and
collected. The purified extract, standards and water were
added to the modified Wade reagent. It was vortexed for
5 seconds and the absorbance was read immediately at
500 nm.

abs of control abs of sample
% inhibition = x 100
abs control
Samples IP6 (g/g) Flavonoids (CE/100 mg) Total phenolics (mg/100 g)
kidney bean 2750.20 9.02
a
145.21 5.03
d
16.38 1.40
a

broad bean 1466.67 15.15
ab
90.61 20.21
d
5.61 1.79
d

Pigeon peas 2483.67 13.21
a
119.91 2.09
d
11.63 0.72
a

Jackfruit seeds 462.50 62.51
c
105.65 34.07
d
22.38 1.73
ab

Pimento 1183.33 16.66
bc
2685.68 15.30
a
2.87 0.17d
Pumpkin seeds (h) 2558.21 18.67
a
60.93 3.21
d
8.23 3.41
d

Pumpkin seeds (u) 2554.67 20.59
a
95.64 24.55
d
21.32 1.57
ab

Corn 2025.52 75.83
a
50.11 2.54
d
80.21 2.14
c

Okra 700.21 17.21
c
595.91 85.53
c
27.95 2.67
b

Sorrel 1520.83 23.52
d
1665.64 18.81
b
5.30 1.30
b

Onion 941.66 16.67
bc
85.86 5.34
d
36.72 1.29
b

Ginger 441.67 25.25
c
470.86 50.34
c
87.99 4.05
c

Golden apple 1945.83 20.83
ab
325.66 35.35
c
28.25 1.70
b

Table 1.0: IP6, Flavoniods and total phenolics in legumes, seeds and spices
Values in the same column with different letter subscripts are significantly different p<0.05. Values are
expressed as mean SEM.
Statistical analyses
Data were finally expressed as means SEM.
Analysis of variance was used to ascertain differences
among different samples by using the Statistical package
for the social sciences software version 16.0.
Differences among means were assessed by the
Duncans multiple range test where significance was
confirmed by a cutoff p value <0.05, (Sokal and Rohlf,
1969).

IP
6

Since IP
6
is found mostly in the aleurone layer of
cereals and grains we would expect highest levels in
grain and seed samples. This was generally observed in
the samples of kidney beans (2750.20 9.02 g/g),
pigeon peas (2483.67 13.21 g/g), broad bean
(1466.67 15.15 g/g), pumpkin seeds (2558.21 18.67
g/g) and corn (2025.52 75.83 g/g) with significantly
higher IP
6
concentration compared to other samples
(Table 1). Golden apple also recorded similar IP
6
content
(1945.83 20.83 g/g) but this was unexpected as
analyses were carried out on the fruit itself and not on the
seed portion. This is of significance as golden apple
(referred to as Jew plum in some countries), is one of the
most commonly consumed fruits in the Pacific and
Tropical regions. Its high IP
6
levels therefore warrant
further investigations since this research suggests that
high IP
6
concentrations may be found in the parts of
foods other than seeds. Jackfruit seeds recorded lower
IP
6
concentrations than other seed samples and this was
unexpected. Bioavailability of minerals from this food
source may therefore be higher than that of other seed
foods, since IP
6
may act as a divalent mineral chelator
especially in low mineral nutrient states. This need to be
further explored since food quality is adversely affected
by low mineral bioavailabity. Pimento and sorrel are
versatile foods as they are used as condiments, spices
and for preparing various drinks. While these samples
had lower IP
6
compared to leguminaceous crops, they
still had appreciable levels that can be exploited for
anticarcinogenic and antioxidant properties. Other spices
including ginger, onion and okra recorded low IP
6

concentrations.
It is important to assess ways in which food
samples with high IP
6
concentrations can be exploited
since this bioactive compound is shown to be effective in
reducing the incidences and complications of numerous
metabolic disorders including hyperlipidaemias, diabetes
mellitus and some cancers (Lee et al., 2007; Lehtihet
et al., 2004; Kumar et al., 2010; Vucenik and
Shamsuddin, 2006). While increased consumption of
these foods are encouraged, purified extracts can also be
prepared and marketed for their reported health benefits
Samples % DPPH Inhibition
*
IC50 (mg/mL)
Ascorbic acid 97.42 0.41
a
0.018
Kidney bean 50.85 0.13
b
0.781
Broad Bean 9.21 2.60
c
8.976
Pigeon Peas 9.17 0.86
c
5.413
Jackfruit seeds 21.01 0.55
d
2.052
Pimento 95.54 0.18
a
0.021
Pumpkin seeds (u) 4.67 0.11
c
8.844
Pumpkin seeds (h) 4.63 0.42
c
7.618
Okra 23.51 4.30
d
2.385
Sorrel 59.52 0.87
b
0.391
Onion 8.67 0.44
c
5.779
Ginger 92.16 0.52
a
0.050
Corn 28.68 0.15
d
1.410
Golden apple 19.93 0.23
d
1.779
Table 2.0: Free radical scavenging activity of
methanolic extracts of legumes seeds and spices
*
The % DPPH inhibition represents the mean SD.
+
IC50 values were calculated based on duplicate analysis
of each plant sample.

as nutraceuticals. This assessment of IP
6
in a wide
variety of beans, seeds condiments and fruits provides us
with new knowledge from which further studies can be
carried out. This work indicates immense potential for
increased crop production along with preparation and
promotion of beneficial nutraceutical products.
It was observed thatfor some samples, IP
6

concentration deviated widely from other reported
values. Differences may however be due to variations in
the assessment methods used since some methods may
measure all phosphate containing compounds within the
sample resulting in the overestimation of IP
6

concentrations.
Bioactive compounds and Antioxidant activity
The DPPH assay is used as an indication of the
free radical scavenging activity of various samples and
as such may identify potentially beneficial antioxidant
components. It measures the ability of the extracts to
donate an H
+
ion to DPPH effectively for reducing it.
Screening foods for bioactive compounds may lead to
the discovery of highly active compounds with
significant health benefits. Secondary metabolites
including flavonoids, IP
6
and total phenolics contribute
to overall antioxidant activity which was assessed by
DPPH inhibition. Pimento and ginger samples (with
values of 95.54 0.18 % and 92.16 0.52 % inhibition)
recorded significantly increased antioxidant activity
compared to other samples with IC50 values comparable
to the ascorbic acid standard (table 2). This observation
is corroborated by other studies (Padmakumari et al.,
2011; Ghasemzadeh et al., 2011). These two food
samples along with others are used widely in various
traditional preparations as treatment for various ailments
including cancers and inflammatory diseases (Tsai et al.,
2005; Marzouk et al., 2007). Data on flavonoid content
of similar foods from the literature is sparse, however
foods with high flavonoid content are reported to have
antioxidant and anti-inflammatory properties and
contribute positively to cardiovascular health (Verena
et al., 2006). The ability of ginger and pimento to reduce
inflammation, among other health benefits may therefore
be due in part to the high levels of flavonoids (which
contribute to total polyphenolic compounds) and other
phytochemicals that contribute to their overall
antioxidant status and reported therapeutic benefits.
Samples of corn and ginger had significantly
higher phenolic content than other samples assessed with
values of 80.21 2.14 mg/100 g and 87.99 4.05
Fig 1. Calcium concentration in legumes, seeds and spices.
Columns with different assigned letter superscripts are significantly
different, (P<0.05). Six sample replicates were used to assess significant
difference among groups.
mg/100 g respectively, while appreciable levels of
polyphenols were also recorded for samples of onion
(36.72 1.29 mg/100 g), okra (27.95 2.67 mg/100 g)
and golden apple (28.25 1.70 mg/100 g) (Table 1). We
therefore theorize that other compounds in addition to
polyphenols may be contributing to antioxidant activity
of some samples since some samples with high
polyphenol concentrations did not show high antioxidant
activities. High values for DPPH inhibition were also
obtained for kidney bean and sorrel samples suggesting
that extracts from these foods are high in antioxidants.
This research suggests that these food samples in
addition to ginger and pimento, may be useful in
lowering the incidences of some inflammatory diseases
since foods that display high antioxidant are shown to be
beneficial in this regard (Wang et al., 2010; Ramadan
et al., 2011).
In light of these results, other plant preparations
with similar therapeutic benefits should be assessed for
overall antioxidant activity with the aim of producing
nutraceutically beneficial and commercially viable
proprietary preparations. Sorrel for example, matures
during the winter months and the calyces of the flower
are traditionally used to prepare a drink following hot
water extraction. The resulting solution which has a deep
red colour is reported to be high in nutrients and
antioxidants and has hypolipidaemic properties (Ochani
and D'Mello, 2009; Bako et al., 2009). Other research
also suggest a role for sorrel in modulating blood
pressure in hypertensive patients, with flavonoids and
other phytochemicals thought to be the beneficial
compounds in this regard (McKay et al., 2010). Our
results show appreciable antioxidant activity and IP
6
in
sorrel samples with only pimento samples having higher
flavonoid concentrations. Further studies should be
conducted and geared at identifying the specific
compound or compounds responsible for the reported
health benefits in this food sample. This data argues well
for continued consumption and study of pimento, ginger
and sorrel with the aim of correlating therapeutic benefits
based on traditional knowledge with scientific data.
Minerals
Pimento samples displayed significantly higher
calcium concentrations than other samples assessed with
8055.31 347.60 mg/Kg as shown in Figure 1. Data
from the literature on mineral content of this spice is
sparse, however this research indicates that with such
high calcium concentrations, pimento seeds are an
Fig 2. Iron concentration in legumes, seeds and spices. Columns with
different assigned letter superscripts are significantly different, (P<0.05). Six
sample replicates were used to assess significant difference among groups.
explorable source of dietary calcium. This may prove
important especially in aging populations in which
calcium availability and assimilation is a problem.
Golden apple samples have displayed high calcium
levels with a value of 2236.48 140.91 mg/Kg, however
the literature reports higher calcium concentrations for
sorrel compared to our data (Glew et al., 2010). Little
data is available from the literature on mineral content of
golden apple samples however the level of minerals
present in this fruit makes it a prime candidate for further
studies. All other samples recorded calcium values of
less than 1000 mg/Kg. Calcium, copper and iron content
of jackfruit seeds are lower than recorded elsewhere,
however higher levels of zinc were found in samples
from this study compared to another recent study (Ocloo
et al., 2010).
Calcium is important for skeletal development
and integrity while also playing key roles in muscle
function and transmission of neuronal impulses.
Adequate intake is therefore recommended throughout
life. Reduced calcium intake is of special concern in
vulnerable populations including the young, the elderly
and in populations with below average food intake. In
addition to supplementing the diet with traditional
calcium sources, increased intake of these high calcium
foods identified by this study is recommended. Overall,
this research shows that in addition to having high
antioxidant activity, sorrel and pimento samples are also
good sources of calcium. Increased utilization of these
foods to supplement the diet will therefore contribute
significantly to satisfy the recommended daily allowance
of 100 mg for calcium.
Samples of sorrel, ginger and pimento had
significantly higher iron content than all other samples
analysed with pimento samples recording the highest
concentrations (Figure 2). Appreciable levels of iron
were also found in the samples of kidney bean, broad
bean and hulled pumpkin seeds. The values recorded for
iron content of pimento were notably higher than
recorded elsewhere, indicating that levels of these
minerals vary with geographical location and cultivation
methods (Aberoumand, 2011).
Iron is an essential micronutrient with adequate
levels needed for preventing anaemia. It also has
important functions in cellular redox reactions. As a
result foods with high levels of this mineral are therefore
highly desirable. High iron content of some samples
analysed make them prime candidates for micronutrient
Fig 3. Copper concentration in legumes, seeds and spices. Columns with
supplementation especially in mineral deficient diets. In
this regard sorrel was shown to be an important
micronutrient source as its addition to cakes as
supplements improved calcium and iron content
significantly (Almana, 2001).
In addition to high iron concentrations in sorrel
(of 64.29 1.06 mg/Kg), ginger (62.84 1.19 mg/Kg),
and pimento samples (75.25 11.68 mg/Kg), we
theorize that iron from these samples may also be readily
available for metabolism owing to relatively low levels
of mineral chelating agents in these samples compared to
legumes and seeds. Further studies assessing in vitro
bioavailability of iron are however needed since not all
forms of iron present in foods are available for
absorption and utilization by the body. This was
highlighted in previous studies where low iron
bioavailability was observed in some tuber samples with
high overall iron content (Dilworth et al., 2007).
Zinc has many important functions including
maintenance of epithelial structures, neuronal
development and immune cell functioning (Haase and
Rink, 2009). It is therefore important that adequate
amounts are ingested since zinc deficiency is thought to
be a widespread but under reported problem (Prasad,
2003). Our research shows that pumpkin seeds are an
excellent source of this micronutrient (43.23 0.62 mg/
Kg) with significantly higher concentration than other
samples assessed (Figure 4). This bears some
significance as in many countries, pumpkin seeds are not
normally consumed but are instead discarded. This work
therefore adds to the growing body of advocating
arguments for increased promotion and processing of
pumpkin seeds, thereby making them suitable for wide
scale consumption. The high zinc content of pumpkin
seeds may also be a reason for its reported positive
effects on prostate health, since adequate zinc is required
for normal prostate functioning and reduced incidences
of prostate cancerspecific mortality (Epstein et al.,
2011). Pigeon peas, jackfruit seeds, okra and sorrel
samples also had high levels of zinc and may also be
useful in this regard. Jackfruit seeds are also not
normally consumed but can be made edible after
cooking. Seeds from both pumpkin and jackfruit samples
which are not normally consumed should therefore be
promoted for their high zinc content. These are dynamic
food samples which can be prepared as snacks,
appetizers or as ingredients in baked products.

Fig 4. Zinc concentration in legumes, seeds and spices. Columns with
The highest copper concentrations were observed
in okra samples with values of 9.09 1.57 mg/Kg
(Figure 3). There were no significant variations in copper
levels in approximately 50% of samples analyzed
however, the levels found in corn, onion and ginger
samples were significantly lower than all other samples
analysed. Copper is important for electron transport and
oxygen transportation and serve as a catalyst to
numerous enzymes, therefore, intake of a small amount
is indicated (RDA of 1.5-3 mg). Most of the food
samples analysed are therefore good sources of dietary
copper.
Although zinc and copper are important from a
nutritional and biochemical standpoint, national food
surveys have revealed marginal to moderately low
contents of both nutrients in the typical American diet
(Ma and Betts, 2000). From a health perspective, this is
significant since there is a direct correlation between the
dietary Zn/Cu ratio and incidence of cardiovascular
diseases (Cabrera et al., 2003). Supplementing the diet
with foods having sufficient zinc and copper should
therefore contribute significantly to the nutritional
efficacy of the typical diet and may lead to reduced
incidences of cardiovascular diseases.
This research which provides information on
mineral contents and other nutritional properties of food
samples consumed frequently and infrequently, argues
well for their increased consumption. The results of this
study bears significance for the food industry, that some
rarely consumed foods and food products e.g. jackfruit
and pumpkin seeds should be incorporated into
mainstream consumption. This could contribute to
enhanced regional food security. Data also showed that
dynamic ways need to be found for increased utilization
of condiments and natural spices in light of their high
nutrient and antioxidant properties.

CONCLUSIONS
This research shows that some food samples
derived from tropical and temperate plants are high in
essential minerals and bioactive compounds. Some
samples displayed high antioxidant activities which may
be a contributory factor to their reported therapeutic
benefits as seen by their extensive use in traditional and
homeopathic medicine. This work indicates that these
foods should be promoted for their health benefits while
further research should be geared at developing
nutraceutical products from them. This work also
provides evidence for increased production, preparation
and consumption of some underutilized highly nutritious
food samples including jackfruit and pumpkin seeds in
order to supplement general or otherwise nutrient poor
diets. Since preserved samples were used in this study,
further comparative work should be carried out with
farm fresh samples.

ACKNOWLEDGEMENTS
The authors are grateful to the Postgraduate
Research and Publications committee at UWI Mona for
providing financial support for this research. Authors are
also indebted to Sannette Hall for her editorial input.

DECLARATION :
The authors declare no conflict of interest.

REFERENCES
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Article Citation:
Dharitri Borgohain and Bhaben Tanti.
Characterization of silica nanoporous structures of freshwater diatom frustules.
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Characterization of silica nanoporous structures of
freshwater diatom frustules
Keywords:
Freshwater diatom, Frustule, Silica, SEM, Geological Survey of India.
ABSTRACT:

A phytoplanktonic unicellular alga known as diatoms belonging to the class
Bacillariophyceae, possess a distinct, highly ornamented siliceous cell wall consisting
of two overlapping halves. Diatoms are found both in marine and freshwater
environment and also in moist habitats. A study was designed to assess and examine
the morphology of diatoms in Chapanala and Jiajuri, two silica rich sites in Nagaon
district of Assam as reported by Geological Survey of India. Samples were collected
from aquatic and semi-aquatic habitats of the study sites and immediately transferred
to Diatom specific Media. The samples were then subjected to acid wash treatment
for detailed microscopic observations. Nanoporous structures of freshwater diatom
frustules have been well characterized through extensive SEM analysis. The prominent
forms include - Pinnularia sp., Navicula sp., Achnanthidium sp., Nitzschia sp. and
Eunotia sp. The SEM micrographs very clearly showed the presence of fine
nanostructure pores, the valve view and distinct raphe of the diatoms. In the present
study, the sizes of nanoporous silica were found in the range of ~60-170 nm under
SEM observations, suggesting the potentiality to use the diatoms in various
nanotechnological applications.
1195-1200 | JRB | 2014 | Vol 3 | No 7
An International
Authors:
Dharitri Borgohain and
Bhaben Tanti*.

Institution:
Gauhati University,
Guwahati - 781014, Assam,
India.

Bhaben Tanti.

Email Id:

Web Address:
Dates:
Received: 07 Jan 2014 Accepted: 29 Jan 2014 Published: 28 Feb 2014
Original Research

INTRODUCTION
Diatoms areeukaryotic, unicellular or colonial
microalgae inhabiting a wide variety of habitats. Diatoms
are microscopic, sizes ranging from 2m to 2mm and
species are classified mostly by the shapes and patterns
of their hard silica parts. The most characteristic feature
of diatoms is their cell wall or exoskeleton which is built
up of amorphous silica. These extremely diverse group
of phytoplankton form the basis of many aquatic food
chains, and are thought to be responsible for upto 25% of
the worlds net primary productivity. The frustules
possess intricate nanoscale features such as pores, ridges,
areoles, spikes and spines imbedded within the periodic
two-dimensional pore arrays. They are the only
organisms known to possess genetic ability to mineralize
amorphous silica into complex structures. Diatoms are
particularly attractive for nanotechnology because they
build their highly symmetric skeletons with a
nanopattern directly in 3D form (Round et al.,1990).
Biomineralize silica cell walls confer the diatoms diverse
and impressive exoskeletal architecture (Montsant et al.,
2005; Bozarth et al., 2009). The diversity of the silica
structures on the diatom cell walls appears to be quite
significant and extends possibilities for their use in nano-
fabrication of a multitude of devices having wide ranging
applications in biochemical analyses, microsensors,
computing and telecommunications, optical devices,
microrobotics, micro batteries etc. (Gordon and
Parkinson, 2005).
Silica sand deposits have been reported by the
Geological Survey of India (GSI) in the Jiajuri and
Chapanala region of Nagaon district of Assam
(Borpuzari, 2012). Jiajuri hill (26

18

0

to 26

19 0

N
latitude and 92

52

55

to 92

54 15

E longitude)
covers an area of 2.9 km
2
and the possible friable
quartzite is about 7.4 million tones. The friable quartzite
deposits of Jiajuri occurs on plateau with undulating
topography. Chapanala is bounded by latitude 26

20

10 N and longitude 92

51

30

E, covering an area of
0.373 km
2
and possible reserve is 3.5 million tones
(Borgohain and Tanti, 2014). No any extensive
investigation has been carried out to characterize the
diatom from these silica rich areas.

Cell collection and culture
Water and semi-aquatic soil samples were
collected from the sampling sites, Chapanala and Jiajuri
on the basis of habitat stratification (Fig.1). The collected
samples were then transferred in the DM (Diatom
Medium) proposed by Beakes et al., (1988). The medium
was standardized with slight modification and the
composition of stock (per 200ml) includes- Ca(NO
3
)
2
.
4H
2
O 4g, KH
2
PO
4
2.48 g, MgSO
4
.7H
2
O - 5 g,
NaHCO
3
3.18 g, EDTAFeNa 0.45g, EDTANa
2

0.45g, H
3
BO
3
0.496g, MnCl
2
.4H
2
O 0.278g, (NH
4
)
6Mo
7
O
24
.4H
2
O 0.20g, Cyanocobalamine - 0.008g,
Thiamine HCl 0.008g, Biotin 0.008g and
Na
2
SiO
3
.9H
2
O 22.8g (Borgohain and Tanti, 2014).
The cultures were kept in a Bio Chemical
Oxygen (BOD) incubator where cultures were allowed to
grow at 3K light and 18-20 C under 50 Mol photons
m
-2
sec
-1
on a 14:10 hr L : D (Complete light : Dark)
cycle (Fluorescent light, FL40S : D National) and were
growing in an exponential phase for 20-22 days. Pure
cultures of diatoms were preserved and maintained on
DM liquid medium and transferred to fresh medium at a
regular interval of 1 month (Gurung et al., 2012; 2013).
Preparation of diatom frustule for microscopic study
The diatom cells were cleaned by acid to remove
the organic matrix present external to the cell wall (Hasle
and Fryxell, 1970). The cleaned frustule valves were
then stored in ethanol to avoid contamination and
bacterial growth. The structural morphology of the
cleaned diatom frustules were examined by Scanning
Electron Microscope JEOL JSM 6360. The cleaned
frustules were partly mounted on brass stubs and coated
Borgohain and Tanti, 2013
with gold for SEM analysis and digital images were
taken using the system.

SEM analysis
The ultra-structure and morphology of nano-
porous silica frustules of the freshwater diatoms were
investigated from the silica rich sites- Chapanala and
Jiajuri of Nagaon district of Assam. The structural
morphology of the acid treated cleaned frustules were
examined by SEM and the images along with their
nanopore sizes are described.
Class: Bacillariophyceae
Order: Naviculales
Family: Pinnulariaceae
Genus: Pinnularia
Fig. 2. showed that valves are linear to linear-
lanceolate with obtusely rounded, subrostrate apices.
Striae chambered and with abrupt transition. The
external proximal raphe ends dilated, bent slightly.
Length of the valve ranges from 30-48m and width
ranges from 5.5-7.5m. From the SEM images, the
diatom was identified as Pinnularia sp. having the
silicon pore sizes of ~81nm.
Order: Bacillariales
Family: Naviculaceae
Fig.1. Map showing the sampling sites (Source: www.mapsofindia.com).

Genus: Navicula
Fig. 3. showed a scanning electron micrograph
(SEM) where, it was observed that the frustules of the
diatom was rhombic-lanceolate with cuneate apices.
Length of the valve ranges from 75.5-90m and width
ranges from 17-20m. From the SEM images, the diatom
was identified to be Navicula sp. The silica nanopores of
this diatom species showed ~63nm in size.
Order: Achnanthales
Family: Achnanthaceae
Genus: Achnanthidium
Fig. 4. showed that frustules are monoraphid,
valves are linear-lanceolate with slightly capitate ends.
Striae usually uniseriate and radiate throughout both
valves. Length of the valve ranges from 6-21m and
width ranges from 1.5-3m. From the SEM images, the
diatom was identified to be Achnanthidium sp. having
silica nanoporous structure of frustule of ~140-160nm.
Family: Bacillariaceae
Genus: Nitzschia
Fig. 5. revealed that the valves are lanceolate
with sides parallel and tapering rapidly at the poles,
terminating with subcapitate apices. Striae barely visible.
Length of the valve ranges from 12-42m and width
ranges from 3.5-4.5m. From the SEM images, the
diatom was identified as Nitzschia sp. having the silicon
pore sizes of ~60-65 nm.
Family: Eunotiaceae
Genus: Eunotia
Figure 2. SEM micrographs of Pinnulariainterrupta(A) Full view (B) detail surface of the valve showing
A
B
Figure 3. SEM micrographs of Naviculabacillum (A) Full view (B) detail surface of the valve showing pores.
A
B
Fig. 6. revealed that the valves are arched
slightly, the dorsal margin convex and narrowing
towards the ends and ventral margin concave. Striae
radiate at apices. Length of the valve ranges from
21-90m and width ranges from 5.6-7.2m. From the
SEM images, the diatom was identified to be Eunotia sp.
which revealed ~150-170 nm of pore sizes.

CONCLUSION
Inspite of immense potentiality of diatoms in
nanoengineering and technology, no any proper scientific
exploration and exploitation of the freshwater diatoms
has been carried out from North-Eastern part of India.
Silica rich soil has a distinctive type of ecological habitat
supporting specific types of diatoms with different type
of features. Diatom frustules display a diversity of
patterns and structures at the nano to millimetre scale. In
this study, we observed very exciting results in case of
Pinnularia, Navicula and Nitzschia species where their
nanoporous silica sizes are less than 100 nm.
Nanoporous silica with less 100 is considered as
excellent materials for wide range of applications in IT
based industries. Further, as these particles are
biologically generated, so they are most stable, cost-
effective and eco-friendly. The two other diatoms
namely, Achnanthidium and Eunotia are also showing
considerable range of nanoporous silica of ~ 150 nm
over their frustules. Their varied geometries and
nanopore sizes offer a wide range of attributes for
exploitation in nanotechnology based industries. The
highly ordered 3D porous silica nanostructures hold a
promising vicinity for the biological fabrication of
Figure 4. SEM micrographs of Achnanthidiumminutissumum (A) Full view (B) detail surface of the valve showing pores.
A
B
A
B
Figure 5. SEM micrographs of Nitzschiapalea (A) Full view (B) detail surface of the valve showing pores.

nanostructured devices and materials from these silica
rich sites. For that, more characterization is needed for
confirmation and authentication.

ACKNOWLEDGEMENT
The author would like to acknowledge UGC-
SAP (Special Assistance Programme) for providing
financial assistance in the form of Basic Scientific
Research (BSR) fellowship to carryout the work.

REFERENCES
Beakes GW, Canter HM and Jaworski GHM. 1988.
Zoospore ultrastructure of Zygorhizidium affluens and
Z. planktonicum, two chytrids parasitizing the diatom
Asterionella formosa. Canadian J Bot.,66(6): 1054-1067.

Borgohain D and Tanti B. 2014. Diversity of
freshwater diatoms from few silica rich habitats of
Assam, India. J. Res. Bio., 4(1): 1162-1173.

Borgohain D and Tanti B. 2014. Seasonal variations of
freshwater diatoms in the silica rich soils of Assam. J.
Res. Plant Sci., 3(1): 242-248.

Borpuzari P. 2012. Ministry to exploit silica reserves in
N-E. The Financial Express, 20 March.

Bozarth A, Maier UG and Zauner S. 2009. Diatoms in
biotechnology: modern tools and applications. App
Microbiol Biotechnol., 82(2): 195-201.

Gordon R and Parkinson J. 2005. Potential roles for
diatomists in nanotechnology. Journal of Nanoscience
and Nanotechnology. 5: 35-40.

Gurung L, Tanti B, Buragohain AK and Borah SP.
2012. Studies on the freshwater diatom diversity in
Deepar Beel, Assam, India. J Assam Sci Soc., 53(2): 1-6.

Gurung L, Buragohain AK, Borah SP and Tanti B.
2013. Freshwater diatom diversity in Deepor Beel a
Ramsar site. J. Res. Plant Sci., 2(2):182-191.

Hasle GR and Fryxell GA. 1970. Diatoms: cleaning
and mounting for light and electron microscopy.
Transactions of the Americans Microscopical Society. 89
(4): 469-474.

Montsant A, Aheshwari U, Bowler C. and Lopez PJ.
2005.Diatomics: towards diatom functional genomics.
Journal of Nanoscience and Nanotechnology. 5: 5-14.

Round FE, Crawford RM and Mann DG. 1990. The
Diatoms: Biology and Morphology of the Genera,
Cambridge University Press. p. 747.
A
B
Figure 6. SEM micrographs of Eunotiasubarcuatioides (A) Full view (B) detail surface of the valve showing pores.

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Article Citation:
Sharma C and Uday Bhan Singh.
Saprobic status and Bioindicators of the river Sutlej.
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Saprobic status and Bioindicators of the river Sutlej
Keywords:
Saprobity, Bioindicators, River Sutlej, Palmer's Algal Index, BOD
ABSTRACT:

Saprobic status and bioindicators of river Sutlej was conducted at (S1) Ropar
Headworks, (S2) downstream after the confluence with BudhaNallah, (S3) Harike
before the confluence with river Beas, (S4) Harike before the confluence with river
Beas. Water samples were collected on the monthly basis for two consecutive years
(November, 2009-October, 2011), on the basis of saprobic classification given by
Sladecek (1973), (S
1
) could be categorized as oligosaprobic, (S
2
) as
polysaprobic, (S
3
) as mesosaprobic, and (S
4
) as meso-polysaprobic. Data on the
Palmer's Algal Index values revealed that S
2
and S
4
were grossly polluted, S
1
was least
polluted, whereas in S
3
, there were chances of medium degree of organic pollution.
Bioindicator organism may have higher frequency index and they are major peak
forming organisms at different stations and in different seasons. The results also
indicate that the bioindicator species may also behave as peak forming organisms and
their abundant depends upon diverse parameters.
1201-1208 | JRB | 2014 | Vol 3 | No 7
An International
Authors:
Sharma C
1
and
Uday Bhan Singh
2
.

Institution:
1. Department of Zoology,
Panjab University,
Chandigarh-160 014, India.

2. Laboratory of Algal
Biology and Diversity,
Panjab University,
Chandigarh-160 014, India.

Uday Bhan Singh.

Email Id:

Web Address:
Dates:
Received: 10 Dec 2013 Accepted: 15 Jan 2014 Published: 14 Mar 2014
Original Research

INTRODUCTION
Planktons are very sensitive to the change in
the environment they inhabit. Any change in the
habitat in terms of tolerance, abundance, diversity
and dominance leads to the change in the plankton
communities (Verma et al., 2012; Sharma et al., 2013;
Jindal et al., 2013). Biological assessment has
emerged as a valuable alternative for aquatic
ecosystems assessments; since planktonic species
are cosmopolitan in distribution and inhabiting
biological communities show the integrated effects
of the environment including water chemistry
(Singh et al., 2013a; Thakur et al., 2013; Singh and
Sharma, 2014). Trivedy (1988) concluded the use of
phytoplanktons for assessing the degree of pollution
of different water bodies. Phytoplankton or
microalgae are diverse group of chlorophyllous
microorganisms with simple nutritional requirements, be
they eukaryotes (for instance, green algae) or
prokaryotes e.g. cyanobacteria (Singh and Ahluwalia,
2013). Nowadays, macrophytes are also considered as
indicators of water quality (Singh et al., 2013b,c). The
change in envir onmental conditi ons and
phytoplankton community further affects the
zooplankton communities which also respond
quickly to changes in environmental quality.
The use of bioindicators to evaluate trophic state
of water bodies, have often been neglected in the contrast
to physical and chemical methods for analysis of water
(Thadeus and Lekinson, 2010). In the present
investigation, the pollution load of river Sutlej was
assessed on basis of bioindicators and saprobic
assessment.
STUDY AREA
The prosperities of Punjab are based on its
river system. The river Sutlej is the easternmost and
longest river of Punjab. It originates near the
Mansarowar Lake in Tibet. It flows west through
deep Himalayan valleys entering India in the
Kinnaur district, the Sutlej enters Punjab near
Nangal, moves on to plains at Ropar, passes
through district Ludhiana. Four stations (S
1
, S
2
, S
3

and S
4
) were set up on the river to collect water
samples.
S
1
: River Sutlej at Ropar Headworks: This is
located at Ropar Headworks (lat. 3059' N; long.
7631' 12"E; alt. 272m above m.s.l.) in Punjab.
S
2
: River Sutlej downstream after the
confluence with Budha Nallah: It is 95 km
downstream S
1
, where Budha Nallah joins river
Sutlej at village Wallipur (lat. 3058' N; long. 75
37' 49"E; alt. 228 above m.s.l.).
S
3
: River Sutlej upstream before the
confluence with East Bein: This is located at
village Lohian before the confluence of East Bein
with river Sutlej (lat. 3107' N; long. 7506'58"E;
alt. 209m above m.s.l.).
S
4
: River Sutlej at Harike before the
confluence with river Beas: It is downstream S
3
after the confluence of East Bein with river Sutlej
and before the confluence of river Beas (lat. 31
08' N; long. 7459' 13"E; alt. 211m above m.s.l.).

The collections were made monthly for a
period of two year i.e. November 2009 -October
2011.
Physico-chemical analysis:
Physico-chemical parameters of the water
were analyzed according to the standard methods
given in Trivedy and Goel (1986) and APHA
(2005).
Biological analysis:
(i) Collection:
For the collection of biota 100 L of water
was sieved through a ring type bolting silk net (24
meshes mm
2
), fitted with a wide mounted glass
bottle. The samples collected were preserved in 4%
Sharma and Singh, 2014
formaldehyde solution on the spot for the counting
of plankton. For living study and identification of
the biota, separate water sample was collected in
the similar manner.
(ii) Identification:
The books consulted for the identification of
phyto- and zooplankton are: Smith (1950),
Edmondson (1959), Hynes (1960), Pennak (1978)
and Kudo (1986).
(iii) Counting of plankton:
Counting of plankton was done with the help
of Sedgwick-Rafter counting cell as per the
procedure given in Wetzel and Likens (2000).
(iv) Saprobic status:
Saprobic condition in the different stretches
of the river Sutlej was determined on the basis of
BOD
5
(organic pollution load) and by the use of
Palmer' s Algal Index (Palmer, 1969).

Saprobic condition in the different stretches
of the river Sutlej was determined on the basis of
BOD
5
(organic pollution load) and by the use of
Palmer' s Algal Index (Palmer, 1969). To
authenticate the relation between saprobes and bio
indicators, we dealt them separately.
Saprobic status in the different stretches of the
river Sutlej
Sanghu et al., (1987) studied the impact of
various human activities on the water quality of
river Ganga at Garhmukteshwar. They reported
high value of BOD (9.15 mg L
1
), indicats pollution
stress in the river. Bhatnagar and Garg (1998)
studied the interrelationship of plankton population
and water quality of river Ghaggar (Sirsa in
Haryana) and concluded that among all the factors
DO and BOD appeared to be more important in
effecting the biotic populations. Kaur and Saxena
(2002) made water pollution studies of river Sutlej
and reported that higher values of BOD (140-242
ppm), and lower values of DO (0.01-3.40 ppm),
alkalinity (253-337 ppm) were due to mixture of
industrial effluents in the river. Kumar et al.,
(2009) assessed the pollution status of river Ganga
at Kanpur. They reported that due to dumping of
huge quantity of sewage and industrial effluents
directly into the river, serious degradation in water
quality with DO reducing to zero level and other
chemical parameters including BOD and COD load
increasing sharply were resulted. Thakur et al.,
(2013) used Palmer' s Algal Species Pollution Index
for rating water quality of three lakes of Himachal
Pradesh.
The monthly fluctuations in the values of
BOD
5
and Palmer's Algal Index have been given in
Table 1.
Monthly average value of BOD (mg L
-1
) was
1.49 0.74 (0.41-2.7), 31.18 06.33 (21.13-40.12),
3.17 0.97 (1.95-4.92) and 21.00 4.29 (15.31-
28.33) in 2009-10, and 1.54 0.59 (0.35-2.48),
22.42 3.92 (16.16-30.15), 2.43 0.81 (1.2-3.65)
and 19.17 3.55 (15.2-25.41) in 2010-11 at S
1
, S
2
,
S
3
and S
4
respectively.
On the basis of saprobic classification
given by Sladecek (1973), Ropar Headworks (S
1
)
could be categorized as oligosaprobic, River Sutlej
at village Wallipur (S
2
) after the confluence of
Budha Nallah as polysaprobic, at village Lohian
before the confluence of East Bein with river
Sutlej (S
3
) as mesosaprobic, and after the
confluence of East Bein with river Sutlej (S
4
) as
meso-polysaprobic.
The monthly average value of Palmer' s
Algal Index was 7 1.37 (5-9), 19 5.63 (13-30),
10 4.33 (417) and 15 2.99 (1120) in 2009-10,
and 5 2.18 (18), 19 4.16 (1024), 8 4.29 (3
16) and 18 5.20 (1027) in 2010-11 at S
1
, S
2
, S
3

and S
4
respectively. Data on the Palmer' s Algal

Index values revealed that S
2
and S
4
was grossly
polluted, S
1
least polluted, whereas S
3
, there were
chances of medium degree of organic pollution.
Bioindicators
Bio-indicators approach, using the responses
of organisms to evaluate trophic state, have often
been neglected in favour of physical and chemical
analysis of water (Thadeus and Lekinson, 2010;
Thakur et al., 2013). Keeping this in view, present
study was conducted on bioindicators of river
Sutlej. On the basis of presence, absence,
abundance and frequency of appearance and
disappearance, the following organisms could be
designated as bioindictors of saprobic status.
Frequency index of peak forming Phytoplankton
at different stations of river Sutlej
At S
1
, diatoms were mainly constituted by
forms like Cymbella affinis (FI 0.50) and
Fragilaria sp. (FI 0.75), Pinnularia sp. (FI 0.75),
Navicula sp. (FI 0.92) and Amphora pediculus (FI
0.54). Chlorococcales was represented by
Pediastrum simplex (FI 0.92), Scenedesmus
abundans (FI 1). Volvocales were Chlamydomonas
sp. (FI 0.75) and Gonium pectorale (FI 0.79).
Zygnematales were Cosmarium sp. (FI 0.46) and
Hydrodictyon sp. (FI 0.46). Euglenophyceae were
Trachelomonas lacustris (FI 0.33), Euglena tuba
(FI 0.83) and Phacus longicauda (FI 0.50).
Cyanophyceae were Oscillatoria subbrevis (FI
1.00), Calothrix sp. (FI 0.42) and Microcystis sp.
(FI 0.75).
At S
2
, diatoms were Synedra ulna (FI 0.79),
Achnanthes sp. (FI 0.67), Navicula cuspidata (FI
0.79) and Nitzschia palea (FI 0.46). Chlorococcales
were constituted by species like Ankistrodesmus
falcatus (FI 0.88), Chlorella vulgaris (FI 0.67) and
Scenedesmus quadricauda (FI 0.79). Volvocales
were Eudorina elegans (FI 0.75) and Pandorina
morum (FI 0.54). Zygnematales were Closterium
acerosum (FI 0.54), Spirogyra sp. (FI 0.71),
Ulothrix sp. (FI 0.50) and Cladophora glomerata
(FI 0.42). Euglenophyceae were Euglena viridis
(FI 0.58), Phacus pleuronectus (FI 0.88) and
Lepocynclis ovum (FI 0.50). Cyanophyceae were
Oscillatoria princeps (FI 0.79), Anabaena sp., (FI
0.50) Arthrospira jenneri (FI 0.58) and Spirulina
gomontii (FI 0.71).
At S
3
, diatoms were Navicula cryptocephala
(FI 0.38), Cymbella sp. (FI 0.0.54), Navicula
cryptocephala (FI 0.42), Gomphonema gracile (FI
0.42) and Syndera ulna (FI 0.38). Chlorococcales
were Scenedesmus quadricauda (FI 0.42),
s. dimorphous (FI 0.63) and Pediastrum tetras (FI
0.63). Volvocales were Chlamydomonas (FI 0.38),
Chlorogonium sp., (FI 0.63) and Eudorina sp. (FI
0.75). Zygnematales were Closterium acerosum (FI
0.92), Cladophora glomerata (FI 0.42), Spirogyra
sp. (FI 0.58) and Zygnema sp. (FI 0.50).
Euglenophyceae were Euglena acus (FI 0.63),
Lepocinclis sp. (FI 0.50), Phacus pleuronectus (FI
0.83) and Trachelomonas sp. (FI 0.38). Blue-greens
were Oscillatoria princeps (FI 0.88), Microsystis
sp. (FI 0.46) and Spirulina gomontii (FI 0.63).
At S
4
,

diatoms were Cymbella ventricosa (FI
0.58), Syndera ulna (FI 0.50), Navicula cuspidata
(FI 0.58) and Melosira varians (FI 0.54), Diatoma
vulgare (FI 0.50) and Navicula cryptocephala
(FI 0.50). Chlorococcales were Ankistrodesmus
falcatus (FI 0.50), Chlorella vulgaris (FI 0.58),
Scenedesmus quadricauda (FI 0. 58) and
Pediastrum tetras (FI 0.71). Volvocales were
Chlorogonium elongatum (FI 0.71), Eudorina
elegans (FI 0.46) and Pleudorina sp. (FI 0.38).
Zygnematales were Closterium acerosum (FI 0.50),
Cladophora glomerata (FI 0.50), Stigeoclonium
tenue (FI 0.38), Spirogyra sp. (FI 0.54) and
Ulothrix sp. (FI 0.29). Euglenophyceae were
Euglena acus (FI 0.67), Lepocynclis ovum
(FI 0.50), Phacus pleuronectus (FI 0.58) and
Trachelomonas sp. (FI 0.38). Blue-green algae were
Oscillatoria princeps (FI 0.67), Phormidium sp. (FI
0.38) and Spirulina gomontii (FI 0.42).
Frequency index of peak forming Zooplankton
at different stations of river Sutlej
At S
1
, Protozoa were Coleps sp. (FI 0.50),
Colpoda sp. (FI 0.50) and Vorticella sp. (FI 0.67)
and Actinophrys sp. (FI 0.46). Rotifera were
Anuraeopsi s sp. (FI 0. 50), Brachi onus
quadridentatus (FI 0.46), B. forficula (FI 0.75),
Monostyla sp. (FI 0.33) and Notholca sp. (FI 0.54).
Copepods were Cyclops viridis (FI 0.83),
Diaptomus gracilis (FI 0.58), Mesocyclops
leuckarti (FI 0.75) and nauplii (FI 1.00).
Cladocerans were Daphnia sp. (FI 0.75), Moina
brachiata (FI 0.58) and Diaphanosoma sarsi
(FI 0.63).
At S
2
, Protozoa were Colpidium sp.
(FI 0.63), Epistylis sp. (FI 0.63) and Aspidisca sp.
(FI 0.46). Rotifera were Brachionus angularis
(FI 0.42), B. calyciflorus (FI 0.71), Asplanchna
brightwelli (FI 0.67), Epiphanes senta (FI 0.67) and
Rotaria rotatoria (FI 0.50). Copepoda were
Cyclops strenus (FI 0.63), Mesocyclops leuckarti
(FI 0.63) and nauplii (FI 0.96). Cladocerans were
Daphnia pulex (FI 0.79) and Chydorus sp.
(FI 0.79).
At S
3
, Protozoa were Colpoda sp. (FI 0.54),
Stylonychia sp. (FI 0.67), Vorticella convallaria
(FI 0.75) and Colpidium sp. (FI 0.92). Rotifera
were Brachionus quadridentatus (FI 0.67),
B. calyciflorus (FI 0.71) and Asplanchna
brightwelli (FI 0.58). Copepoda were Cyclops
leuckarti (FI 0.67), Mesocyclops leuckarti (FI 0.58)
and nauplii (FI 0.92). Cladocerans were Daphnia
sp. (FI 0.67) and Moina brachiata (FI 0.50).
At S
4
, Protozoa were Stylonychia sp. (FI
0.58), Epistylis sp. (FI 0.67) and Colpidium sp. (FI
S
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1

0.71). Rotifera were Brachionus angularis (FI
0.54), B. calyciflorus (FI 0.50), Asplanchna
brightwelli (FI 0.71), Filinia longiseta (FI 0.50)
and Rotaria rotatoria (FI 0.38). Copepoda were
Cyclops brevcornis (FI 0.75), Cyclops strenuus (FI
0.58) Mesocyclops leuckarti (FI 0.83) and nauplii
(FI 0.83). Cladocerans were Daphnia pulex (FI
0.67) and Moina brachiata (FI 0.46).
On the basis of presence, absence,
abundance and frequency of appearance and
disappearance, the following organisms could be
designated as bioindictors of saprobic status.
Oligosaprobic- Phytoplankton:
Anomoenes sp., Amphora sp., Asterionella
sp., Ceratium sp., Cymbella affinis, Closterium sp.,
Dinobryon sp., Euastrum sp., Sorastrum sp.,
Peridinium sp., Meridion sp., Oscillatoria
subbrevis, Pediastrum simplex, Phacus longicauda,
Polybotrya gracilis, Scenedesmus abundance,
Synura sp. , Tet raedron mi ni mum and
Tr achel omonas l acust ri x . Zoopl ankt on:
Actinophrys sp., Anuraeopsis sp., Bosmina
longirostris, Coleps sp., Cyclops bicuspidatus,
Diaptomus gracilis, Daphnia sp., Difflugia sp.,
Keratella procurva, K. tropica, Notholca sp. and
Vorticella sp.
Polysaprobic- Phytoplankton:
Ankistrodesmus falcatus, Chlorella vulgaris,
Closterium acerosum, Cyclotella sp., Cymbella
ventricosa, Euglena viridis, Gomphonema gracile,
Melosira varians, Navicula cryptocephala,
Oscillatoria princeps, Scenedesmus quadricauda,
Lepocinclis ovum and Synedra ulna. Zooplankton:
Aspidisca sp., Asplanchana brightwelli, Brachionus
angularis, B. calyciflorus, Chydorus sp., Colpidium
sp., Epiphanes senta, Epistylis sp., Eucyclops sp.,
Lecane sp. and Stylonychia sp.

CONCLUSION
Based on our results, it has been concluded that
there is a visionable correlation between saprobity and
bioindicators, which is further strengthened by frequency
index. But, it is not mandatory that abundant species may
act as indicator or any indicator organism should be the
peak forming species. This baseline data clearly explains
that, station (S
1
) could be categorized as oligosaprobic,
(S
2
) as polysaprobic, (S
3
) as mesosaprobic, and (S
4
) as
meso-polysaprobic. But these findings are not
appropriate to make a concrete conclusion and it need
more time and diverse parameters along with their
correlations to make an authenticate results, and this is
now open for further studies.

ACKNOWLEDGEMENTS
The authors are thankful to the Chairperson,
Department of Zoology, Panjab University, Chandigarh,
for providing necessary research facilities. One of the
authors (Uday Bhan Singh) thankfully acknowledges the
Council of Scientific and Industrial Research, New
Delhi, for providing financial assistance in the form of
Junior Research Fellowship and Senior Research
Fellowship.

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