Journal of Research in Biology Volume 3 Issue 7

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Razi University of Agricultural Sciences and Natural Resources, Iran

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Panjab University, India

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Dept of Biotechnology, SDM College (Autonomous),

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Centre for biotechnology visva-bharati, India.

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Acharya Institute of Technology, Bangalore.

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Krishna University, India.

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New University of Lisbon, Caparica, Portugal

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Visva-Bharati (a Central University), India

Dr. Preetham Elumalai [Biochemistry and Immunology] Institute for

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Dr. Mrs. Sreeja Lakshmi PV [Biochemistry and Cell Biology] University of Regensburg, Germany

Dr. Alma Rus [Experimental Biology]

University of jaén, Spain.

Dr. Milan S. Stanković [Biology, Plant Science]

University of Kragujevac, Serbia.

Dr. Manoranjan chakraborty [Mycology and plant pathology]

Bishnupur ramananda college, India.

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

http://jresearchbiology.com/documents/RA0387.pdf






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

Jou

rn

al of R

esearch

in

Biology

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/Km2. 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 Gurjar1,

Amol .S. Kumbhar1*,

Jyotirmay Jena1,

Jaya Kumar Yogesh1,

Chittaranjan Dave1,

Ramesh Pratap Singh2,

Ashok Mishra2.

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

Journal of Research in Biology 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 (22°19’ - 22° 30’N

and 77° 56’ - 78° 20’E) covers an area of 1427.87 km2

(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. During this period

39 permanent transects were established in different

habitat types including riparian patches. Each transect

was surveyed thrice by well trained observer between

Gurjar et al., 2013

1087 Journal of Research in Biology (2013) 3(7): 1086-1092

0600–0900 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


Dete

cti

on

Prob

ab

ilit

y

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 /Km2) 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/Km2) 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

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Jathanna et al., 2008

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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. Journal of Research in Biology (2013) 3(7): 1093-1104

Jou

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esearch

in

Biology

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; 4½- 5½ 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:


Dates: Received: 14 Aug 2013 Accepted: 02 Dec 2013 Published: 18 Jan 2014

Journal of Research in Biology

An International Scientific Research Journal

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. Albert’s 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, 9˚20’0’’N, 76˚30’0’’E,

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 3½ (vs. 2½) 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), 3½ scales between lateral

line and anal fin (vs. 4½), 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 4½- 5½ 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 ½ 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

2½ 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), 2½ scales present in

between lateral line and pelvic fin (vs. 3½ 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

4½ 4½ - 5½ 5½ 4 4½ - 5 4 ½ - 5 ½ 5½

7 LL/V

3½ 3½ 3½ 3 3 - 3½ 2 ½ 3½

8 LL/A

3½ 3½ 3½ 3½ 3½ 3 ½ 4½

9 L/TR

5 ½ / 3½ 5 -5½ /3½ 5/4 5 / 3½ 5½ / 4½ 5 ½ / 2½ 5½ / 4½

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. 4½- 5½ 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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Biology

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



www.jresearchbiology.com

Journal of Research in Biology

An International Scientific

Research Journal

Authors:

Kumarasamy D.

Institution:

Department of Botany,

Annamalai University,

Annamalainagar 608 002,

Tamil Nadu, India.


Kumarasamy D.

Email:


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.

Journal of Research in Biology (2013) 3(7): 1105-1110


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 µm (average 24.7 µm) wide, rectangular to

circular. Radial wall pits mostly uniseriate, in some

places it is biseriate, alternate; pits bordered, circular,

contiguous, 12.5 µm in size. Aperture elliptic, crossed,

6.25 µm long and 2.5 µm wide. Tracheids per mm2

are 1599. Late wood tracheids 10.0-23.7 µm (average

11.1 µm) in radial diameter. Rays uniseriate, a few are

partially biseriate, 1-19 (average 6) cells high,

homocellular, cells 22.3 µm long and 17.5 µm wide.

Kumarasamy, 2013


Both tangential and horizontal walls are smooth. Radial

wall pits 3-9, circular, bordered, 7.5 µm wide, tightly

packed. Aperture circular, ray cells spanning 2½-3

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.

100μm a 100μm b

5μm d 50μm c 5μm 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 ep or t ed fr om th i s for ma t i on n am el 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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Kumarasamy, 2013


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190–197.

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of some Upper Gondwana deposits of Palar basin,

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Kumarasamy, 2013



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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. Journal of Research in Biology (2014) 3(7): 1182-1194

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esearch

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Biology

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, IP6, total polyphenols and antioxidant activity. IP6 was determined by anion exchange chromatography while flavonoids, polyophenols, minerals and antioxidant activity were determined by standard methods. Results: The highest concentrations of IP6 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:


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; Pérez-

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 IP6 (also known as phytic

acid or phytate when in salt form), is also thought to play

a role in antioxidant activity of cells. IP6 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). IP6 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


http://en.wikipedia.org/wiki/Inositol

http://en.wikipedia.org/wiki/Salt

http://en.wikipedia.org/wiki/Phosphorus

http://en.wikipedia.org/wiki/Plant

http://en.wikipedia.org/wiki/Biological_tissue

http://en.wikipedia.org/wiki/Bran

http://en.wikipedia.org/wiki/Seed

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 Na2CO3 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. IC50 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.

IP6

Assessment of IP6 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 IP6 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 (µg/g) Flavonoids (CE/100 mg) Total phenolics (mg/100 g)

kidney bean 2750.20 ± 9.02a 145.21 ± 5.03d 16.38 ± 1.40 a

broad bean 1466.67 ± 15.15ab 90.61 ± 20.21d 5.61 ± 1.79d

Pigeon peas 2483.67 ± 13.21a 119.91 ± 2.09d 11.63 ± 0.72 a

Jackfruit seeds 462.50 ± 62.51c 105.65 ± 34.07d 22.38 ± 1.73ab

Pimento 1183.33 ± 16.66bc 2685.68 ± 15.30a 2.87 ± 0.17d

Pumpkin seeds (h) 2558.21 ± 18.67a 60.93 ± 3.21d 8.23 ± 3.41d

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

Corn 2025.52 ± 75.83a 50.11 ± 2.54d 80.21 ± 2.14c

Okra 700.21 ± 17.21c 595.91 ± 85.53c 27.95 ± 2.67b

Sorrel 1520.83 ± 23.52d 1665.64 ± 18.81b 5.30 ± 1.30b

Onion 941.66 ± 16.67bc 85.86 ± 5.34d 36.72 ± 1.29b

Ginger 441.67 ± 25.25c 470.86 ± 50.34c 87.99 ± 4.05c

Golden apple 1945.83 ± 20.83ab 325.66 ± 35.35c 28.25 ± 1.70b

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

Duncan’s multiple range test where significance was

confirmed by a cutoff p value <0.05, (Sokal and Rohlf,

1969).


IP6

Since IP6 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 µg/g),

pigeon peas (2483.67 ± 13.21 µg/g), broad bean

(1466.67 ± 15.15 µg/g), pumpkin seeds (2558.21 ± 18.67

µg/g) and corn (2025.52 ± 75.83 µg/g) with significantly

higher IP6 concentration compared to other samples

(Table 1). Golden apple also recorded similar IP6 content

(1945.83 ± 20.83 µg/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 IP6 levels therefore warrant

further investigations since this research suggests that

high IP6 concentrations may be found in the parts of

foods other than seeds. Jackfruit seeds recorded lower

IP6 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 IP6 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 IP6 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 IP6

concentrations.

It is important to assess ways in which food

samples with high IP6 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 ± 0.41a 0.018

Kidney bean 50.85 ± 0.13b 0.781

Broad Bean 9.21 ± 2.60c 8.976

Pigeon Peas 9.17 ± 0.86c 5.413

Jackfruit seeds 21.01 ± 0.55d 2.052

Pimento 95.54 ± 0.18a 0.021

Pumpkin seeds (u) 4.67 ± 0.11c 8.844

Pumpkin seeds (h) 4.63 ± 0.42c 7.618

Okra 23.51 ± 4.30d 2.385

Sorrel 59.52 ± 0.87 b 0.391

Onion 8.67 ± 0.44 c 5.779

Ginger 92.16 ± 0.52a 0.050

Corn 28.68 ± 0.15d 1.410

Golden apple 19.93 ± 0.23d 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 IP6 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, IP6

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 IP6

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, IP6 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 IP6 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 cancer–specific 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.

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Article Citation: Dharitri Borgohain and Bhaben Tanti. Characterization of silica nanoporous structures of freshwater diatom frustules. Journal of Research in Biology (2014) 3(7): 1195-1200

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Biology

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:


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 2µm 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 world’s 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 km2 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 km2 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(NO3)2.

4H2O – 4g, KH2PO4– 2.48 g, MgSO4.7H2O - 5 g,

NaHCO3 – 3.18 g, EDTAFeNa – 0.45g, EDTANa2 –

0.45g, H3BO3 – 0.496g, MnCl2.4H2O – 0.278g, (NH4)

6Mo7O24.4H2O – 0.20g, Cyanocobalamine - 0.008g,

Thiamine HCl – 0.008g, Biotin – 0.008g and

Na2SiO3.9H2O – 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 µMol photons

m-2sec-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 30-48μm and width

ranges from 5.5-7.5μm. 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 75.5-90μm and width

ranges from 17-20μm. 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 6-21μm and

width ranges from 1.5-3μm. 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 12-42μm and width

ranges from 3.5-4.5μm. 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

21-90μm and width ranges from 5.6-7.2μm. 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. Journal of Research in Biology (2014) 3(7): 1201-1208

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Biology

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), (S1) could be categorized as oligosaprobic, (S2) as polysaprobic, (S3) as mesosaprobic, and (S4) as meso-polysaprobic. Data on the Palmer's Algal Index values revealed that S2 and S4 were grossly polluted, S1 was least polluted, whereas in S3, 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 C1 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:


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 environmental conditions 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 (S1, S2, S3

and S4) were set up on the river to collect water

samples.

S1: River Sutlej at Ropar Headworks: This is

located at Ropar Headworks (lat. 30°59'N; long.

76°31' 12"E; alt. 272m above m.s.l.) in Punjab.

S2: River Sutlej downstream after the

confluence with Budha Nallah: It is 95 km

downstream S1, where Budha Nallah joins river

Sutlej at village Wallipur (lat. 30°58'N; long. 75°

37'49"E; alt. 228 above m.s.l.).

S3: 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. 31°07'N; long. 75°06'58"E;

alt. 209m above m.s.l.).

S4: River Sutlej at Harike before the

confluence with river Beas: It is downstream S3

after the confluence of East Bein with river Sutlej

and before the confluence of river Beas (lat. 31°

08'N; long. 74°59' 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

BOD5 (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

BOD5 (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

BOD5 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 S1, S2,

S3 and S4 respectively.

On the basis of saprobic classification

given by Sladecek (1973), Ropar Headworks (S1)

could be categorized as oligosaprobic, River Sutlej

at village Wallipur (S2) after the confluence of

Budha Nallah as polysaprobic, at village Lohian

before the confluence of East Bein with river

Sutlej (S3) as mesosaprobic, and after the

confluence of East Bein with river Sutlej (S4) 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 (4–17) and 15 ± 2.99 (11–20) in 2009-10,

and 5 ± 2.18 (1–8), 19 ± 4.16 (10–24), 8 ± 4.29 (3–

16) and 18 ± 5.20 (10–27) in 2010-11 at S1, S2, S3

and S4 respectively. Data on the Palmer's Algal



Index values revealed that S2 and S4 was grossly

polluted, S1 least polluted, whereas S3, 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 S1, 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 S2, 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 S3, 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 S4, 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 S1, 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

Anuraeopsis sp. (FI 0.50), Brachionus

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 S2, 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 S3, 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 S4, Protozoa were Stylonychia sp. (FI

0.58), Epistylis sp. (FI 0.67) and Colpidium sp. (FI



Sta

tion

In

dex

Y

ear

Nov.

Dec

. Jan

. F

eb.

Mar.

A

pr.

M

ay.

Ju

n.

Ju

l.

Au

g.

Sep

. O

ct.

S1

Bio

chem

ical

ox

ygen

dem

and

(m

g L

–1)

2009-1

0

0.4

1

0.6

2

0.8

5

1.2

1

1.3

5

1.6

6

2

.41

2

.70

2

.03

2

.30

1

.45

1

.00

2010-1

1

0.3

5

0.9

8

1.2

4

1.4

0

1.6

7

1.8

6

2

.48

2

.13

2

.00

1

.84

1

.63

0

.96

Pal

mer

‟s A

lgal

In

dex

2009-1

0

9.0

0

5.0

0

5.0

0

9.0

0

9.0

0

7.0

0

7

.00

7

.00

7

.00

8

.00

8

.00

8

.00

2010-1

1

8.0

0

4.0

0

4.0

0

4.0

0

7.0

0

7.0

0

7

.00

7

.00

7

.00

1

.00

4

.00

8

.00

S2

Bio

chem

ical

ox

ygen

dem

and

(m

g L

–1)

2009-1

0

24.7

2

21.1

3

25.6

5

29.8

2

31.4

4

38.7

3

34

.41

40

.12

36

.22

38

.34

28

.22

25

.41

2010-1

1

19.2

3

16.1

6.

18.4

3

20.7

2

25.4

1

23.0

0

30

.15

26

.43

24

.72

21

.73

19

.43

17

.42

Pal

mer

‟s A

lgal

In

dex

2009-1

0

22.0

0

16.0

0

13.0

0

14.0

0

19.0

0

22.0

0

24

.00

30

.00

16

.00

10

.00

24

.00

20

.00

2010-1

1

23.0

0

17.0

0

15.0

0

20.0

0

19.0

0

21.0

0

23

.00

24

.00

21

.00

10

.00

22

.00

24

.00

S3

Bio

chem

ical

ox

ygen

dem

and

(m

g L

–1)

2009-1

0

2.0

3

1.9

5

2.2

4

2.4

6

2.8

1

3.6

6

4

.23

4

.92

4

.12

3

.86

3

.13

2

.64

2010-1

1

1.2

0

1.4

4

1.6

5

1.8

3

2.2

0

2.7

5

3

.65

3

.34

3

.32

2

.76

2

.94

2

.13

Pal

mer

‟s A

lgal

In

dex

2009-1

0

6.0

0

5.0

0

4.0

0

6.0

0

11.0

0

11.0

0

11

.00

11

.00

10

.00

15

.00

16

.00

17

.00

2010-1

1

6.0

0

3.0

0

4.0

0

4.0

0

9.0

0

13.0

0

7

.00

7

.00

7

.00

9

.00

15

.00

16

.00

S4

Bio

chem

ical

ox

ygen

dem

and

(m

g L

–1)

2009-1

0

16.2

4

15.3

1

16.6

3

18.9

4

20.8

3

24.2

1

26

.44

28

.33

25

.14

22

.46

19

.63

17

.88

2010-1

1

16.0

5

15.2

0

16.2

1

17.8

9

19.4

6

22.3

7

24

.11

25

.41

22

.12

19

.31

16

.43

15

.55

Pal

mer

‟s A

lgal

In

dex

2009-1

0

14.0

0

11.0

0

11.0

0

16.0

0

20.0

0

15.0

0

16

.00

13

.00

16

.00

14

.00

18

.00

20

.00

2010-1

1

18.0

0

11.0

0

10.0

0

21.0

0

22.0

0

14.0

0

15

.00

19

.00

19

.00

14

.00

24

.00

27

.00

Ta

ble

: 1

Mo

nth

ly f

luctu

ati

on

s in

th

e b

ioch

em

ica

l o

xy

gen

dem

an

d a

nd

Pa

lmer's

alg

al

ind

ex

at

dif

feren

t st

ati

on

s d

urin

g N

ov

em

ber

20

09

to

Octo

ber 2

01

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 . , Te t raedron min imum and

Tr ache lo mon as l acus t r i x . Z oopla nk t 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 (S1) could be categorized as oligosaprobic,

(S2) as polysaprobic, (S3) as mesosaprobic, and (S4) 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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Schernewski G, Neumann T. The trophic state of the Baltic Sea a century ago: a model simulation study. J Mar Sys., 2005;53:109–

124.

Kaufman PD, Cseke LJ, Warber S, Duke JA and Brielman HL. Natural Products from plants. CRC press, Bocaralon, Florida. 1999;

15-16.

Kala CP. Ecology and Conservation of alphine meadows in the valley of flowers national park, Garhwal Himalaya. Ph.D Thesis,

Dehradun: Forest Research Institute, 1998; 75-76.

http://www.ethnobiomed.com/content/pdf/1746-4269-1-11.pdf.

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Journal of Research in Biology Volume 3 Issue 7

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