Journal of Research in Biology Volume 4 Issue 1 by Journal of Research in Biology

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List of Editors of Editors in the Journal of Research in Biology Managing and Executive Editor: Abiya Chelliah [Molecular Biology] Publisher, Journal of Research in Biology. Editorial Board Members: Ciccarese [Molecular Biology] Universita di Bari, Italy. Sathishkumar [Plant Biotechnologist] Bharathiar University. SUGANTHY [Entomologist] TNAU, Coimbatore. Elanchezhyan [Agriculture, Entomology] TNAU, Tirunelveli. Syed Mohsen Hosseini [Forestry & Ecology] Tarbiat Modares University (TMU), Iran. Dr. Ramesh. C. K [Plant Tissue Culture] Sahyadri Science College, Karnataka. Kamal Prasad Acharya [Conservation Biology] Norwegian University of Science and Technology (NTNU), Norway. Dr. Ajay Singh [Zoology] Gorakhpur University, Gorakhpur Dr. T. P. Mall [Ethnobotany and Plant pathoilogy] Kisan PG College, BAHRAICH Ramesh Chandra [Hydrobiology, Zoology] S.S.(P.G.)College, Shahjahanpur, India. Adarsh Pandey [Mycology and Plant Pathology] SS P.G.College, Shahjahanpur, India Hanan El-Sayed Mohamed Abd El-All Osman [Plant Ecology] Al-Azhar university, Egypt Ganga suresh [Microbiology] Sri Ram Nallamani Yadava College of Arts & Sciences, Tenkasi, India. T.P. Mall [Ethnobotany, Plant pathology] Kisan PG College,BAHRAICH, India. Mirza Hasanuzzaman [Agronomy, Weeds, Plant] Sher-e-Bangla Agricultural University, Bangladesh Mukesh Kumar Chaubey [Immunology, Zoology] Mahatma Gandhi Post Graduate College, Gorakhpur, India. N.K. Patel [Plant physiology & Ethno Botany] Sheth M.N.Science College, Patan, India. Kumudben Babulal Patel [Bird, Ecology] Gujarat, India.

Dr. Afreenish Hassan [Microbiology] Department of Pathology, Army Medical College, Rawalpindi, Pakistan. Gurjit Singh [Soil Science] Krishi Vigyan Kendra, Amritsar, Punjab, India. Dr. Marcela Pagano [Mycology] Universidade Federal de São João del-Rei, Brazil. Dr.Amit Baran Sharangi [Horticulture] BCKV (Agri University), West Bengal, INDIA. Dr. Bhargava [Melittopalynology] School of Chemical & Biotechnology, Sastra University, Tamilnadu, INDIA. Dr. Sri Lakshmi Sunitha Merla [Plant Biotechnology] Jawaharlal Technological University, Hyderabad. Dr. Mrs. Kaiser Jamil [Biotechnology] Bhagwan Mahavir Medical Research Centre, Hyderabad, India. Ahmed Mohammed El Naim [Agronomy] University of Kordofan, Elobeid-SUDAN. Dr. Zohair Rahemo [Parasitology] University of Mosul, Mosul,Iraq. Dr. Birendra Kumar [Breeding and Genetic improvement] Central Institute of Medicinal and Aromatic Plants, Lucknow, India. Dr. Sanjay M. Dave [Ornithology and Ecology] Hem. North Gujarat University, Patan. Dr. Nand Lal [Micropropagation Technology Development] C.S.J.M. University, India. Fábio M. da Costa [Biotechnology: Integrated pest control, genetics] Federal University of Rondônia, Brazil. Marcel Avramiuc [Biologist] Stefan cel Mare University of Suceava, Romania. Dr. Meera Srivastava [Hematology , Entomology] Govt. Dungar College, Bikaner. P. Gurusaravanan [Plant Biology ,Plant Biotechnology and Plant Science] School of Life Sciences, Bharathidasan University, India. Dr. Mrs Kavita Sharma [Botany] Arts and commerce girl’s college Raipur (C.G.), India. Suwattana Pruksasri [Enzyme technology, Biochemical Engineering] Silpakorn University, Thailand. Dr.Vishwas Balasaheb Sakhare [Reservoir Fisheries] Yogeshwari Mahavidyalaya, Ambajogai, India.

CHANDRAMOHAN [Biochemist] College of Applied Medical Sciences, King Saud University.

Dr. Pankaj Sah [Environmental Science, Plant Ecology] Higher College of Technology (HCT), Al-Khuwair.

B.C. Behera [Natural product and their Bioprospecting] Agharkar Research Institute, Pune, INDIA.

Dr. Erkan Kalipci [Environmental Engineering] Selcuk University, Turkey.

Kuvalekar Aniket Arun [Biotechnology] Lecturer, Pune.

Dr Gajendra Pandurang Jagtap [Plant Pathology] College of Agriculture, India.

Mohd. Kamil Usmani [Entomology, Insect taxonomy] Aligarh Muslim university, Aligarh, india.

Dr. Arun M. Chilke [Biochemistry, Enzymology, Histochemistry] Shree Shivaji Arts, Commerce & Science College, India.

Dr. Lachhman Das Singla [Veterinary Parasitology] Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, India.

Dr. AC. Tangavelou [Biodiversity, Plant Taxonomy] Bio-Science Research Foundation, India.

Vaclav Vetvicka [Immunomodulators and Breast Cancer] University of Louisville, Kentucky.

Nasroallah Moradi Kor [Animal Science] Razi University of Agricultural Sciences and Natural Resources, Iran

José F. González-Maya [Conservation Biology] Laboratorio de ecología y conservación de fauna Silvestre, Instituto de Ecología, UNAM, México.

T. Badal Singh [plant tissue culture] Panjab University, India

Dr. Kalyan Chakraborti [Agriculture, Pomology, horticulture] AICRP on Sub-Tropical Fruits, Bidhan Chandra Krishi Viswavidyalaya, Kalyani, Nadia, West Bengal, India. Dr. Monanjali Bandyopadhyay [Farmlore, Traditional and indigenous practices, Ethno botany] V. C., Vidyasagar University, Midnapore. M.Sugumaran [Phytochemistry] Adhiparasakthi College of Pharmacy, Melmaruvathur, Kancheepuram District. Prashanth N S [Public health, Medicine] Institute of Public Health, Bangalore. Tariq Aftab Department of Botany, Aligarh Muslim University, Aligarh, India. Manzoor Ahmad Shah Department of Botany, University of Kashmir, Srinagar, India. Syampungani Stephen School of Natural Resources, Copperbelt University, Kitwe, Zambia. Iheanyi Omezuruike OKONKO Department of Biochemistry & Microbiology, Lead City University, Ibadan, Nigeria. Sharangouda Patil Toxicology Laboratory, Bioenergetics & Environmental Sciences Division, National Institue of Animal Nutrition and Physiology (NIANP, ICAR), Adugodi, Bangalore. Jayapal Nandyal, Kurnool, Andrapradesh, India. T.S. Pathan [Aquatic toxicology and Fish biology] Department of Zoology, Kalikadevi Senior College, Shirur, India. Aparna Sarkar [Physiology and biochemistry] Amity Institute of Physiotherapy, Amity campus, Noida, INDIA. Dr. Amit Bandyopadhyay [Sports & Exercise Physiology] Department of Physiology, University of Calcutta, Kolkata, INDIA . Maruthi [Plant Biotechnology] Dept of Biotechnology, SDM College (Autonomous), Ujire Dakshina Kannada, India.

Dr. Satish Ambadas Bhalerao [Environmental Botany] Wilson College, Mumbai Rafael Gomez Kosky [Plant Biotechnology] Instituto de Biotecnología de las Plantas, Universidad Central de Las Villas Eudriano Costa [Aquatic Bioecology] IOUSP - Instituto Oceanográfico da Universidade de São Paulo, Brasil M. Bubesh Guptha [Wildlife Biologist] Wildlife Management Circle (WLMC), India Rajib Roychowdhury [Plant science] Centre for biotechnology visva-bharati, India. Dr. S.M.Gopinath [Environmental Biotechnology] Acharya Institute of Technology, Bangalore. Dr. U.S. Mahadeva Rao [Bio Chemistry] Universiti Sultan Zainal Abidin, Malaysia. Hérida Regina Nunes Salgado [Pharmacist] Unesp - Universidade Estadual Paulista, Brazil Mandava Venkata Basaveswara Rao [Chemistry] Krishna University, India. Dr. Mostafa Mohamed Rady [Agricultural Sciences] Fayoum University, Egypt. Dr. Hazim Jabbar Shah Ali [Poultry Science] College of Agriculture, University of Baghdad , Iraq. Danial Kahrizi [Plant Biotechnology, Plant Breeding,Genetics] Agronomy and Plant Breeding Dept., Razi University, Iran Dr. Houhun LI [Systematics of Microlepidoptera, Zoogeography, Coevolution, Forest protection] College of Life Sciences, Nankai University, China. María de la Concepción García Aguilar [Biology] Center for Scientific Research and Higher Education of Ensenada, B. C., Mexico Fernando Reboredo [Archaeobotany, Forestry, Ecophysiology] New University of Lisbon, Caparica, Portugal Dr. Pritam Chattopadhyay [Agricultural Biotech, Food Biotech, Plant Biotech] Visva-Bharati (a Central University), India

Veeranna [Biotechnology] Dept of Biotechnology, SDM College (Autonomous), Ujire Dakshina Kannada, India.

Dr. Preetham Elumalai [Biochemistry and Immunology] Institute for Immunology Uniklinikum, Regensburg, Germany

RAVI [Biotechnology & Bioinformatics] Department of Botany, Government Arts College, Coimbatore, India.

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

Sadanand Mallappa Yamakanamardi [Zoology] Department of Zoology, University of Mysore, Mysore, India.

Dr. Alma Rus [Experimental Biology] University of jaén, Spain.

Anoop Das [Ornithologist] Research Department of Zoology, MES Mampad College, Kerala, India.

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 4 - Issue 1) Serial No

Accession No

Title of the article

Page No

RA0410

Diversity of freshwater diatoms from few silica rich habitats of Assam,

1162-1173

India. Dharitri Borgohain and Bhaben Tanti.

RA0395

Detection of biofilm formation in urinary isolates: need of the hour.

1174-1181

Saha R, Arora S, Das S, Gupta C, Maroof KA, Singh NP and Kaur IR.

RA0415

Foraging and pollination behavior of Apis mellifera adansonii Latreille

1209-1219

(Hymenoptera: Apidae) on Glycine max L. (Fabaceae) flowers at Maroua. Fernand-Nestor Tchuenguem Fohouo and Dounia. 4

RA0421

Determining the Natural Gypsophila L. (Coven) Taxa Growing in Tunceli

1220-1227

(Turkey). Mustafa Korkmaz and Hasan Ozรงelik.

RA0422

Distribution pattern of birds in Banni Grassland of Kachchh district,

1228-1239

Gujarat, India Mukesh H. Koladiya, ArunKumar Roy Mahato, Nikunj B. Gajera and Yatin S. Patel. 6

RA0414

Determination of age and growth by scale of a population of common trout (Salmo trutta macrostigma, Dumeril, 1858) at the level of Sidi Rachid River (Ifrane. Morocco). Abba H, Belghity D, Benabid M and Chillasse L.

1240-1246

Journal of Research in Biology

An International Scientific Research Journal

Original Research

Journal of Research in Biology

Diversity of freshwater diatoms from few silica rich habitats of Assam, India Authors: Dharitri Borgohain and Bhaben Tanti*.

Institution: Department of Botany, Gauhati University, Guwahati - 781014, Assam, India.

ABSTRACT: Diatoms are a ubiquitous class of phytoplankton of extreme importance for the biogeochemical cycling of minerals such as silica. Few places of Nagaon district of Assam, India viz., Jiajuri, Borhola, Thanajuri and Chapanala have been recognized as the highest silica zones by Geological Survey of India. No any research has been conducted to explore the diatom diversity at this important silica rich habitat. In the present investigation, the morphology and diversity of freshwater diatom species were investigated during May 2012 to April 2013. The samples were subjected to acid wash treatment followed by microscopic observations. Altogether 103 species of diatoms belonging to 20 genera were recorded. Occurrence of diatom varied in all the four different study sites. The dominant genera includes: Stauroneis, Kobayasiella, Eunotia, Pinnularia, Nitzschia, Gomphonema, Frustulia, Surirella, Achnanthes, Rhopalodia, Navicula, Synendra, Encyonema, Achnanthidium, Cymbella, Hippodonta, Tabularia, Actinella, Encyonopsis and Luticola. Notably, all the diatom species belonged to pennate type.

Corresponding author: Bhaben Tanti.

Keywords: Freshwater diatoms, silica rich soil, diatom diversity, Geological Survey of India.

Email Id:

Article Citation: Dharitri Borgohain and Bhaben Tanti. Diversity of freshwater diatoms from few silica rich habitats of Assam, India. Journal of Research in Biology (2014) 4(1): 1162-1173

Web Address:

http://jresearchbiology.com/ documents/RA0410.pdf.

Dates: Received: 07 Jan 2014

Accepted: 29 Jan 2014

Published: 15 Feb 2014

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. Journal of Research in Biology An International Scientific Research Journal

1162-1173 | JRB | 2014 | Vol 4 | No 1

www.jresearchbiology.com

Borgohain and Tanti, 2014 (26°20′10″ N latitude and 92°51′30″ E longitude) deposits

INTRODUCTION Diatoms belonging to the class Bacillariophyceae

occur friable quartzite covering an area of 0.373 km 2 and

are the major group of single-celled photosynthetic

possible reserve is 3.5 million tones. Thanajuri hill

eukaryotic algae which can be found in almost all

(26°12′ 35’’ to 26°13′10″ N latitude 92°48′40″ to 92°50′35″

aqueous and humid environments. Diatoms are an

E longitude and) is situated in the northern part of Karbi-

important component of phytoplankton in freshwaters.

Anglong plateau and southern part of Nagaon district.

There are over 250 genera of diatoms with more than

The possible reserves of glass sand is about 1.788

100,000 species (Gurung et al., 2012, Van Den Hoek

million tones. Friable quartzite occurs in Borhola (26°26′

et al., 1997), which includes both marine and the

15″ N latitude and 92°56′45″ E longitude) covering an

freshwater environments. These microscopic autotrophic

area of 0.595 km2 and the possible reserve of glass sand

microalgae

wall

is about 1.25 million tones. Till date, there is no any

composed of glass silica (SiO2) called frustules which

extensive work on the detailed investigation of diatom

provide a variety of shapes from nano to micro-scale

diversity in these silica rich regions of Assam. Set in this

structures. Diatoms can occur in large amounts, either

backdrop, the present investigation is assessed for the

solitary or in colony and is cosmopolitan in distribution.

exploration of diatom, having the genetic ability to

A major constituent of the plankton family, diatoms are

deposit natural silica over their cell surface in

free floating, planktonic or attached to a substrate and

characteristics nanoporous forms.

possess

highly ornamented

cell

benthic forms (Werner, 1977). Diatoms are important from the point of the biogeochemical cycling of silica.

MATERIALS AND METHODS

Diatoms play a very significant ecological role by fixing

Sample collection and growth conditions

about 25% carbon globally. The diatoms of North East

Samples were collected from aquatic and semi-

region of India are still largely unexplored and

aquatic habitats of the four study sites- Jiajuri, Borhola,

unexploited. Friable quartzite’s belonging to the Shillong

Thanajuri and Chapanala from May 2012 to April 2013

groups of rocks occur sporadically along eastern most

(Fig.1). The freshly collected samples were immediately

part of the Nagaon district. Borhola, Chapanala, Jiajuri

transferred to Diatom Medium (DM) proposed by

and Thanajuri are some of the important places where

Beakes et al., (1988) which was standardized with slight

friable quartzites are found abundantly. About 75% of

modifications and the composition of stock (per 200ml)

the glass sand may be recovered from this friable

includes- Ca(NO3)2. 4H2O – 4g, KH2PO4– 2.48 g,

quartzite by using different methods of beneficiation

MgSO4.7H2O - 5 g, NaHCO3- 3.18 g, EDTAFeNa-

(Goswami, 2006).

0.45g, EDTANa2 – 0.45g, H3BO3 – 0.496g, MnCl2.

The Geological Survey of India (GSI) has found

4H2O

–0.278g,

(NH4)

6Mo7O24.4H2O

–

0.20g,

significant reserves of silica deposits in the Jiajuri region

Cyanocobalamine - 0.008g, Thiamine HCl – 0.008g,

between the district of Nagaon and Karbi Anglong in

Biotin – 0.008g and Na2SiO3.9H2O – 22.8g.

Assam (Borpuzari, 2012). The area is located about

One ml of each stock solution was added to make

30kms South-East from Nagaon and is adjacent to Jiajuri

the final volume of 1L with distilled water, and adjusted

′ ″

Tea Estate. The deposit is bounded by latitude 26° 18 0

to pH 6.8. For solid medium, 1.5% agar was added. The

to 26°19′ 0″ N and longitude 92°52′ 55″ to 92°54′ 15″ E.

cultures were allowed to grow at 3K light at 18-20°C for

Jiajuri hill covers an area of 2.9 km and the possible

20-22 days. Repeated sub-cultures were done on the

friable quartzite is about 7.4 million tones. Chapanala

solid medium to obtain pure cultures of diatom species.

1163

Journal of Research in Biology (2014) 4(1): 1162-1173

Borgohain and Tanti, 2014

Figure 1: Map showing the four study areas (source: www.mapsofindia.com). Cleaning diatom frustules by acid wash method for

until the cell suspension become less acidic. To confirm

microscopic analysis

the complete removal of organic matters, a drop of

In order to analyze the diatom frustules for

cleaned samples was observed under the microscope.

microscopic studies, a cleaning procedure was needed

For light microscopy (LM) observation, the

that removed the external organic matrix covering the

slides were prepared by evaporating drops of the cleaned

frustules. Plankton samples were subjected to acid wash

diatoms suspended in distilled water onto cover-slips and

method according to the protocol of Hasle and Fryxell

the mounting was done by using Naphrax (a specific

(1970) before light microscopic observations. About

diatom mountant with refractive index 1.74). The slides

20ml of liquid cultures were transferred into a beaker and

were examined carefully under 1000x magnification and

treated with equal quantity of concentrated H2SO4 and

the diatom images were documented in Nikon ECLIPSE

agitated gently. Freshly prepared KMnO4 was added to

E200 with photo micrographic attachment.

the sample until the sample had a purple tint. Then

Identification of diatoms

freshly prepared oxalic acid (COOH)2 was added to

The diatoms obtained through laboratory pure

obtain clear solution. The sample was centrifuged at

cultures were identified by consulting various literatures

2500 rpm for 15 min and then rinsed with distilled water

and monographs (Gandhi, 1955; Husted, 1959; Hendey,

Journal of Research in Biology (2014) 4(1): 1162-1173

1164

Borgohain and Tanti, 2014 1964; Patrick and Reimer 1966; Prescott, 1975; Desikachary, 1989; Round et al., 1990; Nautiyal et al., 1996; Anand, 1998; Gurung et al., 2013).

culture were enumerated. Out of 103 diatoms species obtained in pure cultures, 25 diatoms were found to be of different species of Nitzschia representing 24.3% of the total diatom flora. Further, there were 17 different species of Gomphonema,

RESULTS AND DISCUSSION During the present investigation, a total of 103

15 different species of Navicula, 14 different species of

species of freshwater diatoms belonging to 20 genera of

Pinnularia

class Bacillariophyceae were reported from the silica rich

representing

soils of Nagaon district of Assam i.e. Jiajuri, Borhola,

respectively. There were four different species of

Thanajuri and Chapanala. The prominent genera in

Stauroneis, Cymbella (3.9% each), followed by Frustulia

terms of its abundance and frequency were Nitzschia

and Synendra (2.9% each) and Achnanthes and

(25), Gomphonema (17), Navicula (15), Pinnularia (14),

Achnanthidium (1.9% each). The remaining diatoms viz.

Eunotia (5), Stauroneis (4), Cymbella (4), Frustulia (3),

Surirella,

Synendra (3), Achnanthes (2), Achnanthidium (2) and

Encyonopsis, Rhopalodia, Luticola, Hippodonta and

single species of the following diatoms: Actinella,

Kobayasiella were represented by only one species

Luticola, Encyonema, Hippodonta, Surirella, Tabularia,

showing 8.7% out of the total diatoms identified in pure

Encyonopsis,

cultures (Fig. 2).

Kobayasiella

and

Rhopalodia.

Pure

cultures of diatoms obtained in this study were identified

and

different

species

14.6%,

13.6%

16.5%,

Tabularia,

Encyonema,

of and

Eunotia 4.9%

Actinella,

Taxonomic account:

upto their genus level (Fig. 3-9). Morphological

Taxonomic description of the 20 pennate

descriptions of the diatom isolates obtained in pure

freshwater diatom genera obtained in the four silica rich

Diversity of diatom flora

Figure 2: Representation of diatom flora diversity. 1165

Journal of Research in Biology (2014) 4(1):1162-1173

Borgohain and Tanti, 2014 sites during the study period are described below:

coarse, 2-4 middle striae short and thick, radiate in the

Class: Bacillariophyceae

middle, convergent towards apices.

Order: Bacillariales

Class: Bacillariophyceae

Family: Naviculaceae

Order: Cymbellales

Genus: Navicula Bory 1822, Cleve 1894

Family: Gomphonemataceae

Navicula sp. (Fig. 3 A-O)

Genus: Gomphonema C.A. Agardh 1824

Valves 36 µm long, 14 µm broad, broadly

Gomphonema sp. (Fig. 5 A-L, 6 M-Q)

elliptical with convex margins; ends slightly produced,

Valves 45 µm long and 8 µm broad, clavate with

slightly capitate rounded; raphe thin, straight; central

capitate head pole and slightly capitate foot pole; axial

nodules distinct; axial area narrow, linear; central area

area linear, narrow, and widening into a small circular

somewhat obliquely rectangular; striae 23 in 10 µm, very

central area with an isolated pore on the primary side of

fine.

the central nodule; raphe straight with distinct central

Class: Bacillariophyceae

nodules; striae 10-11 in 10 µm, punctate and slightly

Order: Naviculales

radiate, wider at the centre of the valve.

Family: Pinnulariaceae

Class: Bacillariophyceae

Genus: Pinnularia Ehrenberg 1843

Order: Naviculales

Pinnularia sp. (Fig. 4 A-N)

Family: Amphipleuraceae

Valves 53 µm long, 11 µm broad, linear, more or less parallel margins with slightly tapering, broadly rounded ends; raphe thick, straight, placed on one side

Genus: Frustulia Lange-Bertalot Frustulia sp. (Fig. 6 A-C) Valves

71-160 µm long and 15.3-30.2 µm

with distinct, unilaterally curved central nodules and

broad, rhombic-lanceolate, narrowing sharply to the

curved terminal fissures; axial area distinct, linear;

rounded apices. Axial and central areas narrow but

central area large reaching the sides; striae 7 in 10 µm,

distinct. Transverse striae perpendicular to the raphe at

B F

J O L

Figure 3(A-O): Navicula. Journal of Research in Biology (2014) 4(1): 1162-1173

1166

Borgohain and Tanti, 2014

Figure 4: (A-N) Pinnularia the center of the valve, sometimes becoming slightly

Family: Diadesmidaceae

convergent towards the ends of the valve, but radiate at

Genus: Luticola (Ehrenberg) D. G. Mann, 1990

the apices, striae 20-30 in 10 µm.

Luticola sp. (Fig. 6 E)

Class: Bacillariophyceae

Valves 12-24 µm long and 7-9 µm broad, linear

Order: Cymbellales

to linear-elliptical. Transapical striae radiate throughout,

Family: Cymbellaceae

composed of two to four rounded areolae. Largest

Genus: Encyonema (Berkeley) Kutzing

areolae near the valve margins. One isolated, circular

Encyonema sp. (Fig. 6 D)

stigma present, striae 18-20 in 10 µm.

Valves 37-91 µm long and 15-30 µm broad,

Class: Bacillariophyceae

robust and broadly dorsiventral and symmetrical to the

Order: Cymbellales

transapical axis. Dorsal margin normally arched, ventral

Family: Cymbellaceae

margin biarcuate to convex. Valve apices bluntly

Genus: Encyonopsis (Grunow) Krammer, 1997

rounded. Raphe straight with central endings deflected

Encyonopsis sp. (Fig. 6 F)

dorsally and apical ends deflected ventrally, striae coarse

Valves 21-25 µm long and 5.1-6.3 µm broad,

and 8-21 in 10 µm.

cymbelloid with dorsal margin strongly curved and

Class: Bacillariophyceae

straight ventral margin. Axial area narrow, straight and

Order: Naviculales

without a central area. Small central nodule. A stigmoid

1167

Journal of Research in Biology (2014) 4(1):1162-1173

Borgohain and Tanti, 2014

D E

J K

Figure 5 (A-L):Gomphonema. Presented near the dorsal central striae, striae 14.2-16 in

Class: Bacillariophyceae

10 µm.

Order: Eunotiales

Class: Bacillariophyceae

Family: Eunotiaceae

Order: Rhopalodiales

Genus: Actinella Lewis, 1864

Family: Rhopalodiaceae

Actinella sp. (Fig. 7 C)

Genus: Rhopalodia Otto Muller, 1895: 57 Rhopalodia sp. (Fig. 7 A)

Valves 76-140 µm long and 5.7-8 µm broad, arcuate, asymmetrical to both the apical and transapical

Valves 21-30 µm long and 6-9 µm broad,

axes. External distal raphe ends extending slightly to the

isopolar and dorsiventral, lanceolate-elliptical in shape,

valve face on both ends. Striae parallel, striae 13-19 in

acute apices. The dorsal margin curved and straight at

10 µm.

the ventral margin. Striae composed of a single row of

Class: Bacillariophyceae

puncta composes. Fibulae radiate, striae 14-20 in 10 µm.

Order: Achnanthales

Class: Bacillariophyceae

Family: Achnanthaceae

Order: Naviculales

Genus: Achnanthidium Kutzing, 1844

Family: Naviculaceae

Achnanthidium sp. (Fig. 7 D and E)

Genus: Kobayasiella Lange-Bertalot, 1999 Kobayasiella sp. (Fig. 7 B)

Valves 6.2-14 µm long and 2-3.7 µm broad, linear-elliptic, slightly or more elongated near the end,

Valves 22-26 µm long and 5-7 µm broad, linear-

and with bluntly rounded poles. Striae slightly radiate

lanceolate with convex sides and short, capitate apices.

and often a shortened striae near the small central area,

The axial area is narrow and nearly linear. The central

axial area narrow, striae 19-21 in 10 µm.

area is small and elliptical and bordered by alternately

Class: Bacillariophyceae

long and short striae, striae 35-40 in 10 µm.

Order: Bacillariales

Journal of Research in Biology (2014) 4(1): 1162-1173

1168

Borgohain and Tanti, 2014

Figure 6: (A-C) Frustulia, D– Encyonema, E-Luticola, F-Encyonopsis, (M-Q) Gomphonema. Family: Eunotiaceae

and distantly placed, striae 13 in 10 µm.

Genus: Eunotia Ehrenberg 1837

Class: Bacillariophyceae

Eunotia sp. (Fig.7 F-J)

Order: Bacillariales

Valves 68µm long, 12 µm broad, slightly arched,

Family: Bacillariaceae

dorsal margin convex with two wavy ridges at the

Genus: Nitzschia Hassall, 1845: 435

middle, gradually narrowing towards the ends, ventral

Nitzschia sp. (Fig. 8 A-Y)

margin concave; ends slightly constricted on the dorsal

Valves 27-30 µm long and 5.2-6.7 µm broad,

side, slightly produced, rounded; raphe thin; polar

linear with concave sides and wedge shaped, constricted

nodules distinct, on the ventral side near the apices;

produced ends, striae very fine, almost indistinct, striae

striae 13 in 10µm, coarse, lineate, parallel, somewhat

31-35 in 10 µm.

radiate and closely placed near apices.

Class: Bacillariophyceae

Order: Naviculales

Order: Fragilariales

Family: Naviculaceae

Family: Fragilariaceae

Genus: Hippodonta (Ehrenberg)

Genus: Synendra Ehrenberg 1832: 87

Hippodonta sp. (Fig. 9 A)

Synendra sp. (Fig. 7 K-M)

Valves 20.2-29 µm long and 5.5-8 µm broad,

Valves 44 µm long and 3.2- 3.8 µm broad, linear

elliptic-lanceolate, ends subcapitate to capitate. Raphe

with narrow and capitate ends. The central area reaches

straight, filiform, central pores fairly close. Striae

the margins. Pseudo raphe linear and broad. Striae strong 1169

Journal of Research in Biology (2014) 4(1): 1162-1173

Borgohain and Tanti, 2014

Figure 7 A: Rhopalodia, B- Kobayasiella, C- Actinella, D and E- Achnanthidium, (F-J) Eunotia, (K-M) Synendra. noticeably broad, radiate in the middle, convergent at the

Class: Bacillariophyceae

ends, striae 9-11 in 10 µm.

Order: Fragilariales

Class: Bacillariophyceae

Family: Fragilariaceae

Order: Surirellales

Genus: Tabularia (C. Agardh) D.M. Williams and

Family: Surirellaceae

Round

Genus: Surirella Turpin 1828

Tabularia sp. (Fig. 9 E)

Surirella sp. (Fig. 9 B)

Valves 21-400 µm long and 3.1-5.3 µm broad,

Valves 55-65 µm long and 30-34 µm broad,

elliptic or elongate and variable in outline, from narrowly

heteropolar, ovate with broad rounded ends. Middle line

linear to linear- lanceolate or lanceolate valves with

absent. Middle field linear-lanceolate. Striae very thick,

rounded or capitate ends, striae 7.4-25 in 10 µm.

widening towards the middle, set at unequal distances,

Class: Bacillariophyceae

Striae 11-16 in 10 µm.

Order: Cymbellales

Class: Bacillariophyceae

Family: Cymbellaceae

Order: Achnanthales

Genus: Cymbella, C.A. Agardh 1830

Family: Achnanthaceae

Cymbella sp. (Fig. 9 F-I)

Genus: Achnanthes C.A. Agardh (1824) Achnanthes sp. (Fig. 9 C & D)

Valves 118 µm long, 24 µm broad, ventricose, curved, asymmetric, dorsal side convex, ventral side

Valves 12.5-16 µm long and 5-7 µm broad,

slightly concave with middle inflation; ends slightly

rectangular-elliptical to almost quadrate in the middle

constricted, produced rounded; raphe thick, arcuate,

portion, constricted at the ends which are rostrate. Axial

excentric with ventrally curved central nodules; axial

area narrow and central area linear reaching the margins.

area not narrow; central area elliptical with 3-4 isolated

Journal of Research in Biology (2014) 4(1): 1162-1173

1170

Borgohain and Tanti, 2014

Figure 8(A-R):Nitzschia

Figure 8(S-Y):Nitzschia stigmata at the ends of the middle ventral striae; striae

central pores and curved terminal fissures. Axial area

8-10 in 10 µm, punctate, radiate.

moderate, linear or slightly widened between the middle

Class: Bacillariophyceae

and ends: Striae radial, striae 20-22 in 10 µm.

Order: Naviculales

It is interesting to note that all the diatom taxa

Family: Stauroneidaceae

belonged to pennate type. No centric forms of diatom

Genus: Stauroneis Ehrenberg, 1843

were found in all the four sampling sites. Majority of the

Stauroneis sp. (Fig. 9 J-M)

forms were solitary and colonial forms were absent. The

Valves 62-66 µm long and 15-18 µm broad,

dominant genera includes- Gomphonema, Nitzschia,

lanceolate with abruptly constricted, somewhat produced

Stauroneis, Navicula, Frustulia, Eunotia and Pinnularia

capitate ends. Raphe thick with slightly unilaterally bent

which were common in all the sampling sites in all the

1171

Journal of Research in Biology (2014) 4(1): 1162-1173

Borgohain and Tanti, 2014

Figure 9. A- Hippodonta, B- Surirella, C and D- Achnanthes, E- Tabularia, (F-I) Cymbella, (J-M) Stauroneis. seasons throughout the year. Kobayasiella, Cymbella,

ACKNOWLEDGEMENT

Synendra, Achnanthidium and Tabularia were abundant

The author would like to acknowledge UGC-

only in Chapanala while Luticola, Encyonema occurred

SAP (Special Assistance Programme) for providing

Basic Scientific Research (BSR) fellowship in carrying

Borhola.

Pennate

diatoms

Achnanthes,

Encyonopsis, Hippodonta, Actinella and Rhopalodia

out the work.

were found only in Jiajuri. Only pennate diatom Surirella was found in Thanajuri.

REFERENCES Anand N. 1998. Indian fresh water microalgae. Bishen

CONCLUSION Silica rich soils Jiajuri, Borhola, Thanajuri and Chapanala of Nagaon district of Assam harbours rich assemblage of various forms of diatoms; many of which are new to the region. As detailed taxonomic investigations on the diatom flora of North- East India is very limited, the present basic information of diversity

Singh Mahendra Pal Singh Publication, Dehradun, India. p. 1-94. 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.

and distribution of diatoms would form a useful tool for

Borpuzari P. 2012. Ministry to exploit silica reserves in

further monitoring and ecological assessment of these

N-E India. The Financial Express 20 March.

silica rich soils of Assam. Further, the diversity of freshwater diatoms could also be used as a resource database for future applications.

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Desikachary TV. 1989. Atlas of

Diatoms: Marine

diatoms of the Indian Ocean region. Madras Science Foundation. 6(1-13): 622-809.

1172

Borgohain and Tanti, 2014 Gandhi HP. 1955. A contribution to our knowledge of

Van Den Hoek C, Mann DG and Jahns HM. 1997.

fresh-water diatoms of Pratapgarh, Rajasthan. J. Indian

Algae: An introduction to phycology, Cambridge

Bot Soc., 34(4): 307-338.

University Press, London.

Goswami ID. 2006. Mineral resources of Assam, In

Werner D. 1977. The Biology of Diatoms. University of

Envis Assam, July-September, p. 2-4.

California Press. p. 498.

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 Deepar Beel â&#x20AC;&#x201C; 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. Hendey NI. 1964. An introductory account of the smaller algae of British coastal water, Part V, Bacillariophyceae (Diatoms). H.M.S.O., London. 317323. Husted

1959.

Die Kieselalgen Deutschlands,

Osterreichs Und Der Schweiz, Vol.2. Koeltz Scientific Books, USA. p. 845. Nautiyal R, Nautiyal P and Singh HR. 1996. Pennate diatom flora of a cold water mountain river, Alaknanda II suborder Araphideae. Phykos. 35(1-2): 57-63. Patrick R and Reimer CW. 1966. The diatoms d Hawaii.I. Monograph of the Acad. Nat. Sci. Philad. 13

Submit your articles online at www.jresearchbiology.com

(1): 668-672.

Advantages

Prescott GW. 1975. Algae of the Western Great Lakes Area. Michigan State University, USA. p. 998-1012. Round FE, Crawford RM and Mann DG. 1990. The

Easy online submission Complete Peer review Affordable Charges Quick processing Extensive indexing You retain your copyright

diatoms: biology and morphology of the genera,

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Journal of Research in Biology (2014) 4(1): 1162-1173

Journal of Research in Biology

An International Scientific Research Journal

Original Research

Journal of Research in Biology

Detection of biofilm formation in urinary isolates: need of the hour Authors: ABSTRACT: Saha R1*, Arora S1, Das S1, Gupta C1, Maroof KA2, The purpose of the study was to estimate biofilm (BF) formation in urinary Singh NP1 and Kaur IR1. catheterized patients, by comparing three methods i.e. Tissue culture plate method (TCP), Congo Red Agar method (CRM) and Tube method (TM) and to study the antimicrobial resistance pattern in BF producing and non BF producing isolates. A total Institution: of 130 urinary catheterized patients were taken as the study group. From one milli 1. Department of litre of urine sample isolates > 102 colony forming units per milli litre were screened Microbiology, University College of Medical Sciences for the detection of BF by TCP, TM and CRM. Antibiotic sensitivity test for both BF producing and non BF producing bacterial and fungal isolates were done as per CLSI and Guru Teg Bahadur guidelines. From 130 urine samples in our study group, 55 samples grew Hospital, Dilshad Garden, microorganisms of significance, of which 11 samples were poly-microbial in nature. Of Delhi â&#x20AC;&#x201C; 110095, India. these biofilm production was seen in 49 microorganisms (89.09%) by any of the three methods used. TCP method picked up 69% of biofilm producers as compared to TM 2. Department of and CRM which picked up only 36% and 27% biofilm producers respectively. Our study Community Medicine, University College of reveals TCP method as the more dependable one as compared to TM and CRA Medical Sciences and Guru methods for the quantitative biofilm detection, so it can be recommended as a Teg Bahadur Hospital, screening method in laboratories. Dilshad Garden, Delhi â&#x20AC;&#x201C; 110095, India. Keywords: Biofilm, biofilm detection, Congo Red Agar. Corresponding author: Rumpa Saha.

Abbreviations BF - Biofilms; TCP - Tissue Culture Plate; CRM - Congo Red Method; TM - Tube Method; CLSI - Clinical Laboratory Standard Institute; CAUTI - Catheter associated Urinary Tract Infection; CLED - Cysteine Lactose Electrolyte Deficient; BHIB - Brain Heart Infusion Broth; TSB - Trypticase soy broth; ELISA - Enzyme linked immunosorbent assay; MHA - Muller Hinton Agar; MIC -Minimum Inhibitory Concentration; ATCC - American type culture collection; GPC -Gram positive cocci; GNB - Gram negative bacilli.

Web Address:

Article Citation: Saha R, Arora S, Das S, Gupta C, Maroof KA, Singh NP and Kaur IR. Detection of biofilm formation in urinary isolates: need of the hour. Journal of Research in Biology (2014) 4(1): 1174-1181

http://jresearchbiology.com/ documents/RA0395.pdf.

Dates: Received: 01 Dec 2013

Accepted: 08 Feb 2014

Published: 17 Feb 2014

1174-1181 | JRB | 2014 | Vol 4 | No 1

www.jresearchbiology.com

Saha et al., 2014 patient to become incontinent, thus leading to failure of

INTRODUCTION Indwelling urinary catheters play an essential

medical device.

part in the management of disorders of the urinary tract,

There are different methods for the estimation of

especially in the elderly and disabled patients. These

biofilm formation including Tissue culture plate method,

urinary catheters serve as a portal of entry for

Tube method, Congo Red agar method, bioluminescent

microorganisms leading to Catheter Associated Urinary

assay, light or fluorescence microscopic examination,

Tract Infections (CAUTI). Many of these microbes

confocal laser scanning microscope and piezoelectric

colonize and adhere to the artificial surface of the

sensor (Mathur et al., 2006).

indwelling catheters, which then forms biofilms.

There is paucity of data in Indian literature

Biofilms are communities of microorganisms which are

regarding biofilm formation in urinary catheterized

embedded within a matrix of extracellular polymeric

patients. This study was undertaken with the aim to

material and display an altered phenotype. Based on the

estimate biofilm formation in urinary catheterized

type and length of the stay of a gadget, composition of

patients, to compare three methods i.e. Tissue culture

microorganism in a biofilm may vary from one to

plate method (TCP), Congo Red method (CRM) and

numerous. The same is true for urinary catheter biofilms

Tube method (TM) for biofilm production and to study

where number of organisms is directly proportional to

antimicrobial resistance pattern in biofilm producing

length of exposure.

isolates.

Microorganisms indwelling

urinary

commonly

catheters

are

isolated

from

Staphylococcus

MATERIALS AND METHODS

epidermidis, Escherichia coli, Klebsiella pneumoniae,

The study was done over a period of one year

Enterococcus faecalis, Proteus mirabilis and Candida sp

from April 2008 â&#x20AC;&#x201C; March 2009 at department of

(Donlan, 2001).

Microbiology, of our tertiary care hospital after obtaining

Biofilms carry important clinical repercussions

clearance from Institutional Ethical Committee. A total

as they provide a niche for survival of microbes, by

of 130 urinary catheterized patients were taken as study

conferring

drying,

group who gave informed consent to the work. One ml of

mechanical damage and other influences from external

urine samples were collected from catheter with aseptic

environment, human immune system and antimicrobial

precautions and the samples were immediately sent to

agents (Costerton et al., 1995; Mah and Toole, 2001).

the Microbiology laboratory. The samples were plated on

High antimicrobial concentrations are required to

Cysteine Lactose Electrolyte Deficient (CLED) medium.

inactivate organisms growing in biofilms and resistance

The age, sex, days of catheterization of the patients were

may often increases thousand folds. (Stewart and

noted.

Costerton, 2001)

microbiological procedures. The presence of > 102 c.f.u./

protection

microbes

from

Isolates

were

identified

standard

Moreover biofilms act as a persistent source of

ml in aseptically collected urine was taken as significant

infection or may provide reservoir for new infections.

bacteriuria (Winn et al., 2006). The cultures were

The biofilms often leads to crystalline material blocking

maintained on nutrient agar slopes, Enterococci were

the catheters and induce complications like painful

maintained on brain heart infusion slopes and Candida

distension of the bladder, urolithiasis, reflux of infected

species were maintained on Sabouraudâ&#x20AC;&#x2122;s Dextrose Agar

urine resulting in pyelonephritis and sometimes urinary

(SDA) slopes.

leakage around the outside of the catheter causing the

production in the study were: S. epidermidis ATCC

1175

Control

strains used for

biofilm

Journal of Research in Biology (2014) 4(1): 1174-1181

Saha et al., 2014 35984 (strong biofilm producer), S. epidermidis ATCC

of bacteria adhering to surface and forming biofilms.

35983 (moderate biofilm producer) and S. epidermidis

Experiments were performed in triplicate. Interpretation

ATCC 12228 (non biofilm producer), Acinetobacter

of biofilm production was done according to the criteria

baumannii ATCC 19606 and Candida albicans ATCC

of Stepanovie et al., (2007). (Table 1)

90028.

Tube method: Biofilm formation was detected by the following

A quantitative method was used as described by

three methods:-

Christensen et al., (1982). Ten milli litre of BHI broth

Tissue culture plate method (Christensen et al., 1995):

with 1% w/v glucose was taken in test tubes and was

Isolates from freshly subcultured plates were

inoculated with loop full of microorganism from

inoculated in trypticase soy broth (TSB) with 1% w/v

overnight culture plates and incubated at 37˚C for 24 hrs.

glucose and incubated for 18 hours at 37˚C in stationary

The tubes were washed with PBS (pH 7.3) after

conditions and then diluted to 1:100 with fresh TSB.

decanting the culture. The dried tubes were then stained

Individual wells of sterile polystyrene 96 well flat

with crystal violet (0.1% w/v) for 30 minutes after fixing

bottom microtitre plates were filled with 200μl aliquots

with sodium acetate (2% w/v) for 10 minutes. Through

of diluted culture. Un-inoculated TSB served as a control

washing was again done with de-ionized water to remove

to check sterility and non specific binding of media.

excess stain. Tubes were then kept in inverted position

Control strains were also inoculated in triplicate. The

for complete drying. Biofilm formation was detected by

microtitre plate was incubated for 24 hrs at 37˚C. After

the presence of visible film on the wall and bottom of the

incubation contents of each well was removed by tapping

tube. Ring formation at the liquid culture interface was

the plates. After washing the wells for four times with

taken as negative. The amount of biofilm formation was

200μl of phosphate buffer saline (PBS pH 7.2), the

scored according to the results of control strains and

floating planktonic bacteria were removed. The biofilms

graded as 0, 1, 2 and 3 denoting absent, weak, moderate

thus formed in plates were fixed using 2% w/v sodium

and strong biofilm formation respectively. Experiments

acetate for 10 minutes and tainted with 0.1% w/v crystal

were performed in triplicate.

violet for 30 minutes. After washing thoroughly with de-

Congo red agar method (Freeman et al., 1989):

ionized water to remove any excess stain, the plates were

Congo red media was prepared as a concentrated

dried. Micro-ELISA auto-reader at the wavelength of

aqueous solution of 0.8 g/l of Congo red and autoclaved

540 nm was used to measure the Optical Density (OD) of

separately from other medium constituents [brain heart

the stained adherent micro-organisms. The OD540 value

infusion broth (37 g/l), sucrose (50 g/l), agar (10 g/l)];

of sterile medium, fixative and dye were averaged and

then added when agar gets cooled to 55˚C. The required

subtracted from all test values. The mean OD540 value

microbial strains were inoculated on the prepared media

from a control well was deducted from all test OD540

and incubated aerobically for 24 hrs at 37˚C. Growth of

values. These OD540 values were considered as an index

black colonies with a dry crystalline consistency was

Table 1. Interpretation of biofilm production Average OD value

Biofilm production

≤ OD540C/ OD540C < ~ ≤ 2x OD540C

Non/weak

2x OD540C < ~ ≤ 4x OD540C

Moderate

> 4x OD540C

Strong

Journal of Research in Biology (2014) 4(1): 1174-1181

taken as positive biofilm production; pink colonies with occasional darkening at the centre of the colonies were non biofilm producers. Black colonies without dry crystalline colonial morphology indicated indefinite results. The experiment was performed in triplicate and repeated for three times. 1176

Saha et al., 2014 Table 2. Comparison of biofilm production by three methods – TCP, TM and CRM Isolate Gram positive organism n-12 Staphylococcus aureus n = 8 Enterococcus sp n=4 Gram negative organism n-37 Escherichia coli n = 20 Klebsiella sp n=7 Citobacter sp n=2 Proteus sp n=2 Acinetobacter sp n=2 Pseudomonas sp n=4 Candida sp n-6 Candida albicans n=2 Candida tropicalis n=4 Total n = 55

TCP (%) 11(91.66) 7 4 24(64.86) 13 4 1 1 2 3 3 (50) 1 2

TM (%) 2(16.66) 1 1 17(45.94) 11 3 0 1 1 1 1(16.66) 0 1

CRM (%) 2 (16.66) 2 0 12 (32.43) 5 3 1 2 0 1 1 (16.66) 0 1

No BF producer (%) 1 (8.33) 1 0 4 (10.81) 3 1 0 0 0 0 1 (16.66) 1 0

38(69.09)

20(36.36)

15 (27.27)

6 (10.90)

Antimicrobial susceptibility testing was done

RESULTS

on Muller-Hinton agar (MHA) for both biofilm

Among 130 urine samples from our study group,

producing and non biofilm producing bacterial isolates

55 samples grew microorganisms of significance of

by Kirby Bauer disk diffusion method as per Clinical and

which 11 samples were polymicrobial in nature. Of these

Laboratory Standards Institute guidelines (CLSI, 2006).

biofilm production was seen in 49 microorganisms

The antibiotic panels used were 25μg Cotrimoxazole,

(89.09%) by any of the three methods used. All sets of

30μg

units

polymicrobial organisms were biofilm producers. All

Nitrofurantoin, 10μg Norfloxacin, 120μg High level

comparisons were done keeping TCP as gold standard.

gentamicin, 30μg Tetracycline, 30μg Amikacin, 10μg

The different organism isolated and their biofilm

Gentamicin, 10μg Imipenam, 100μg Piperacillin; 10μg

producing capacity is compared in Table 2.

Cefotaxime,

30μg

Vancomycin,

300

Tazobactam and 300 units Polymyxin B . Antibiotics

TCP method picked up 69% (38) of biofilm

discs were procured from HiMedia Laboratories Pvt. Ltd,

producers as compared to TM and CRM which picked up

India.

only 36% (20) and 27% (15) of biofilm producers Antifungal susceptibility profile of BF forming

respectively. This difference was found to be highly

and non biofilms forming Candida isolates was done by

significant (x2 = 17.55, P < 0.001). Table 3 shows

determining MIC for Amphotericin B, Itraconazole and

sensitivity and specificity of TM and CRM. By TCP

Fluconazole by microdilution method as described by

method, the number of strong biofilm producers were 20

CLSI guidelines (CLSI, 2008). Candida albicans ATCC 90028 were used as control. Statistical Analysis: Data entered in MS Excel and SSPS 17.0 were used for data analysis. Chi square test was used to compare

proportions

between

various

groups.

Sensitivity, Specificity and predictive values were calculated using the standard formulae. 1177

Table 3. Diagnostic parameters TM and CRM for Biofilm detection Parameters

CRM

Sensitivity

34.21%

21.05%

Specificity

58.82%

Positive Predictive Value

65.00%

53.33%

Negative Predictive Value

28.57%

25.00%

Journal of Research in Biology (2014) 4(1): 1174-1181

Saha et al., 2014 Table 4. Comparison of antimicrobial resistance pattern of BF producer with non BFproducers Antimicrobial agents Staphylococcus aureus n =8 Cotrimoxazole Cefotaxime Vancomycin Nitrofurantoin Norfloxacin Enterococcus n-4 Vancomycin High level Gentamicin Nitrofurantoin Norfloxacin Tetracycline Gram negative organism n=33 Amikacin Gentamicin Cotrimoxazole Imipenam Piperacillin-Tazobactam Norfloxacin Nitrofurantoin Pseudomonas n = 5 Amikacin Gentamicin Imipenam Piperacillin-Tazobactam Polymyxin B Norfloxacin Candida spp n = 6 Fluconazole Itraconazole Amphotericin B

BF producer (%) n= 7 6(85.71) 5(71.42) 0 3(42.86) 6(85.71) n=4 1 (25) 4(100) 2 (50) 4(100) 4(100) n= 21 15 (71.43) 15 (71.43) 18 (85.71) 7 (33.33) 15 (71.43) 17 (80.95) 13 (61.90) n=4 3 (75) 3 (75) 3 (75) 2 (50) 0 3 (75) n=3 2 (66.67) 3 (100) 0

Non BF producer (%) n=1 1 (100) 1 (100) 0 0 0 n=0 n = 12 6 (50) 6 (50) 6 (50) 1 (8.33) 5 (41.67) 8 (66.67) 4 (33.33) n=1 1 (100) 1 (100) 0 0 0 0 n=3 1 (33.33) 2 (66.67) 0

and the same by TM and CRM was 3 and 14 respectively

however it was not significant except for Cotrimoxazole

and this difference was found to be highly significant

(x2 = 4.911, P = 0.0266).

(x2 = 21.4, P < 0.001, d.f = 2). (Figure1). When degree

Biofilm

production

has

also

increased

of biofilm production was compared, TM showed similar

significantly with the days of catheterization (x2 = 16.88,

detection rate with TCP for moderate biofilm producers,

P < 0.001) (Figure 3).

but the same is not true for strong biofilm producers. This difference was also highly significant.(x2 = 21.06, P < 0.001, d.f = 1). Figure 2 shows colonies of biofilm and non biofilm producers on Congo Red medium.

DISCUSSION More than 40% of all healthcare associated infections are due to CAUTI. Eradication of biofilm

The antimicrobial resistance pattern of the

based catheter related infection is often challenging

biofilm producing isolates is given in Table 4. Among

because they exhibit increased resistance to antimicrobial

the gram negative organism, the resistance was more for

therapies by various mechanisms (Douglas, 2003).

biofilm producers as compared to non biofilm producers Journal of Research in Biology (2014) 4(1): 1174-1181

1178

Saha et al., 2014

Figure 1 Degree of biofilm formation by TCP, TM and CRM In this study we evaluated 55 isolates by three

CRM picked up greater number of biofilm producers

different screening methods for their ability to form

among the Gram negative bacilli (GNB). This difference

biofilms. In our study we have found that TCP method

was however not significant (x2 = 197, P = 0.1226,

detected biofilm formation in 69% of isolates. We have

d.f = 2).

used 1% sucrose in BHI for growing biofilms in

TM detected 36% of isolates as biofilm

microtitre plate. Addition of sugar increases the biofilm

producers while 63% isolates were identified as non

production; as reported by other authors (Mathur

biofilm producers. TM is only 34.21% sensitive, 58.82%

et al.,2006; Bose et al., 2009 ; Hassan et al., 2011).

specific for biofilm detection. This is not consistent with

Overall TCP method detected maximum biofilm

the findings of Mathur et al., 2006; Bose et al., 2009

producers. The ability to detect biofilm production of

from India, who reported higher sensitivity and

Gram Positive Cocci (GPC) was less for TM and CRM

specificity for Tube method. In our study, this method

method as compared to TCP method whereas TM and

correlated well with TCP for identifying moderate biofilm producers (30.90% i.e. 17 / 55), but detection rate for high biofilm producer was very low (5.45% i.e. 3/55). This difference may be due to the inter-observer variability in the reading of results, resulting in low sensitivity and specificity in our study. Only 27% isolates were identified as biofilm producers by CRM similar to Ruzicka et al., 2004 who detected 43.5% of biofilm producers by this method. This was higher in comparison to the 3-6% detection rate by other workers from India and Pakistan (Mathur et al., 2006; Bose et al., 2009; Hassan et al., 2011). The sensitivity and specificity, however, remained low

Figure 2. Colonies of biofilm and non biofilm producers on Congo Red agar medium 1179

(21.05% and 58.82% respectively). Surprisingly, in this study CRM outscores TM in the detection of high Journal of Research in Biology (2014) 4(1): 1174-1181

Saha et al., 2014 CONCLUSIONS The ability of microorganisms to form biofilms on the medical devices is a challenge for the clinicians because biofilm associated microorganisms are much more resistant to antimicrobial agents, which may result in treatment failure. Therefore effective treatment strategies should be explored to deal such infections. Our findings indicate that TCP is a suitable and reproducible

Days of catheterization

method for the screening of biofilm producers in health

Figure 3. Relationship of Biofilm production with duration of catheterization.

care setups.

biofilm producers. CRM detected 25.45% (14/55) while

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TM detected 5.45% isolates as high biofilm producers

Bose S, Khodke M, Basak S and Mallick SK. 2009.

and this difference was highly significant. CRM is a

Detection of biofilm producing staphylococci: need of

comparatively easier method and also over-rules

the hour. J Clin Diagn Res., 3(6):1915-1920.

inconsistency by observation which could possibly explain such finding. The antimicrobial

susceptibility pattern

microbes isolated from clinical samples has important implications especially in clinical settings as it helps

Christensen GD, Simpson WA, Bisno AL and Beachey EH. 1982. Adherence of slime-producing strains of Staphylococcus epidermidis to smooth surfaces. Infect Immun. 37(1): 318-26.

clinicians to decide treatment protocol for patients and

Christensen GD, Simpson WA, Younger JJ, Baddour

also help hospital infection control team to formulate

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Journal of Research in Biology

An International Scientific Research Journal

Original Research

Journal of Research in Biology

Foraging and pollination behavior of Apis mellifera adansonii Latreille (Hymenoptera: Apidae) on Glycine max L. (Fabaceae) flowers at Maroua Authors: Fernand-Nestor Tchuenguem Fohouo1 and Dounia1-2*.

Institution: 1. Laboratory of Zoology, Faculty of Science, University of Ngaoundéré, Ngaoundéré, Cameroon. 2. Laboratory of Zoology, Higher Teacher Training College, University of Yaoundé I, Yaoundé, Cameroon.

ABSTRACT: To assess the impact of Apis mellifera adansonii on pod and seed yields of Glycine max, its foraging and pollinating activities were studied in Maroua, during the two season seasons (August-September 2010 and 2011). Observations were made on 51 to 17866 flowers per treatment. Treatment 1 represented by free flowers; treatment 2 bagged flowers and treatment 3 flowers visited only by A. m. adansonii. In addition, all flower visitors were recorded. The abundance of bee, duration of visits, impact of activity of A. m. adansonii on fruiting percentage, the influence of this bee on formation of pods, number of seeds in each pods and average of normal seeds (well developed) were recorded. Individuals from 28 species of insects were recorded on the flowers of G. max, after two years of observations. Apis mellifera adansonii with 23.18% of 954 visits was the most frequent, followed by Polyrachis sp. 1 (14.77%), Macronomia vulpina (14.22%), Lipotriches collaris (11.07%). This honey bee intensely and exclusively foraged for nectar. The mean foraging speed was 12.56 ± 5.79 flowers per minute. Flowers visited by insects had higher fruiting rate compared with the others while those bagged had the lowest. Apis mellifera adansonii foraging resulted to a significant increment in fruiting rate by 14.14 and 11.98%, as well as the number of seeds per pod by 36.95 and 35.65%, and the percentage of normal seeds by 32.61 and 29.26% respectively in 2010 and 2011. The installation of A. m. adansonii colonies in G. max plantations is recommended to improve pod and seeds production of this species.

Corresponding author: Dounia.

Keywords: Apis mellifera adansonii, Glycine max, flower, visit, nectar, pollination.

Email Id:

Article Citation: Fernand-Nestor Tchuenguem Fohouo and Dounia. Foraging and pollination behavior of Apis mellifera adansonii Latreille (Hymenoptera: Apidae) on Glycine max L. (Fabaceae) flowers at Maroua. Journal of Research in Biology (2014) 4(1): 1209-1219

Web Address:

Dates: Received: 15 Jan 2014

http://jresearchbiology.com/ documents/RA0415.pdf.

Journal of Research in Biology An International Scientific Research Journal

Accepted: 04 Feb 2014

Published: 11 April 2014

1209-1219 | JRB | 2014 | Vol 4 | No 1

www.jresearchbiology.com

Fohouo and Dounia, 2014 INTRODUCTION

MATERIALS AND METHODS

Glycine max is an annual plant originated from

Study site, experimental plot and biological material

Northern and Central regions of China (Hymowitz, 1970).

The experimental is carried, from June to October,

The plant is an annual, herbaceous, erect, and can reach a

in 2010 and 2011 at Ma ye l - I bb é (Latitude 10° 62' N,

height of 1.5m; there are cultivars of soybean

Longitude 14°33' E and altitude 400 m), Maroua, Far

indeterminate, determinate and semi-determinate growth

North Region of Cameroon. This Region belongs to the

(Gallais and Bannerot, 1992).

The first leaves are

Savanna zone, with unimodal rainfall (Letouzey, 1985).

simple, opposite and swallowed, while the following are

It has a Sahel-Sudanian climate type, characterized by

trifoliate and alternate; the pod is straight or slightly

two seasons: a more extended dry season (November to

curved, with a length of two to seven cm; the seed is

May) and a brief rainy season (June to October) (Kuete

generally oval, but may vary depending on the cultivar,

almost spherical, elongated and flattened (Hymowitz and

temperature are 1100mm and 38°C respectively

Harlan, 1983). Flowers are grouped by two to eight on a

(Kuete et al., 1993). The experimental plot was 28m x

short racemes inserted on the stem axile sheets and are

5m. The biological material was represented by Apis

purple or white (Boyeldieu, 1991). Each flower has a

mellifera adansonii Latreille (Hymenoptera:

tubular calyx of five sepals, a corolla of five petals, a

and others insects present in the environment. Seed of

single carpel and ten stamens, nine of which being

G. max was provided by the Institute of Agricultural

welded and the tenth is free (Hymowitz and Harlan,

Research for Development (IARD).

1983). Each flowers Produce nectar and pollen which

Sowing and weeding

al.,

1993).

The

maximum

rainfall

and

Apidae),

attract insects (Milfont et al., 2013). The reproduction

On t h e June 12, 2010 and June 15, 2011, the

system is autogam/allogam (Ibarra-Perez et al., 1999).

experimental plot was cleaned and divided into 24

Soybean is grown primarily for its seeds, which have

subplots, each measuring 1m × 1m. Sowing and weeding

many uses in the food and industrial sectors (USDA,

was done as described by Douka and Tchuenguem

2002). It is a major edible oil and vegetable sources of

(2013).

protein (38-40%) for the feed of men and other animals

Determination of the reproduction system of Glycine

(Boyeldieu, 1991; Tien et al., 2002; USDA, 2002).

max

Currently the production of G. max in Cameroon is low

On July 22, 2010, eight subplots carrying 106

whereas the demand for seeds is high (MINADER,

plants with 34395 flowers at the bud stage were labeled.

2010). Therefore, it is important to investigate on the

Four subplots carrying 80 plants with 17187 flowers

possibilities of increasing the production of this plant

were left to be open pollinated (treatment 1) (figure 1)

in the country. This can be done if flowering insects of

and four subplots carrying 17208 flowers were

G. max in each region are well known and exploited

protected with gauze mesh prevent to insect or other

(Milfont et al., 2013). Unfortunately no research has

pollinating animals vi s i t s (treatment 2) (figure 2).

been reported on the relationships between G. max and

On July 28, 2011, the experiment was repeated, for

its anthophilous insects in Cameroon. In Maroua A. m.

treatment 1 four subplots carrying 80 plants with 17866

adansonii visit flowers of G. Max (unpublished data), and

flowers and four treatment 2 four subplots carrying 80

this study is carried out to assess the effects of foraging

plants with 15875 flowers.

activities of A. m. adansonii on yields of G. max.

Twenty days after shading of the last flower, the number of pods was assessed in each treatment.

1210

Journal of Research in Biology (2014) 4(1): 1209-1219

Fohouo and Dounia, 2014

Figure 1. Glycine max plot showing unprotected plants in bloom. The podding index (Pi) was then calculated as described

Figure 2. Glycine max plot showing isolated plants in bloom. (Tchuenguem, 2005).

by Tchuenguem et al., (2004): Pi = F2/F1, where F2 is

During the same time that A. m. adansonii

the number of pods formed and F1 the number of

encountered on flowers were registered, the types of

viable flowers initially set.

floral products collected by this bee were noted. This

The allogamy rate (Alr) from which derives the

parameter was measured to determine if A. m. adansonii

autogamy rate (Atr) was expressed as the difference in

is strictly a pollenivore, nectarivore or pollenivore and

podding indexes between treatment 1 (unprotected

nectarivore. This could give an idea on its implication as

flowers) and treatment 2 ( bagged flowers) as follows

a cross pollinator of G. max.

(Demarly, 1977):

In the morning of each day, the number of

Alr = [(Pi1 - Pi2) / Pi1] × 100, Where Pi1 and Pi2

opened flowers was counted. The determination of

are respectively the podding average indexes of

frequency of visits, the duration of A. m. adansonii on

treatments I and II. Atr = 100 – Alr.

the flower of G. max was recorded according to

Study of the foraging activity of Apis mellifera

Tchuenguem (2005). The number of pollinated visits,

adansonii on Glycine max flowers

the abundance of foragers, the number of flowers

The frequency of A. m. adansonii in the flowers

visited by A. m. adansonii per minute was recording

of G. max was determined based observations on

every day of observation. The method of observation was

flowers of treatments 1 in 2010 and 2011. Experience

followed as given by Tchuenguem et al., (2004).

were made on 17187 individual opened pollinated

The foraging speed was calculated according to

flowers (treatment 1) each day, from July 26 to August

Jacob – Renacle (1989) by this formula: Vb = (Fi/di) x

20, 2010 and from August 2, to August 2 4 , 2011 at

60 where di is the time (s) given by a stopwatch and Fi is

7 – 8 h, 9 – 10 h, 11 – 12 h, 13 – 14 h, 15 – 16 h and 17

the number of flowers visited during di.

– 18 h. Capture and determination of insects that visited

int era cti on bet ween A. m. adansonii and the

G. max flowers was realize as described by Borror and

competitors and the attractiveness exerted by the flower

White (1991).

of other plant species around the experimental plot on

The

The determination of the relative frequency of all

A. m. adansonii were recorded (Tchuenguem et al.,

insects visit the G. max flowers was calculated

2004). The climatic factor (temperature and humidity)

Journal of Research in Biology (2014) 4(1): 1209-1219

1211

Fohouo and Dounia, 2014 was registered as described by Douka and Tchuenguem

seeds (well developed) was then calculated for each

(2013).

treatment 3.

Evaluation of the impact of Apis mellifera adansonii

Data analysis Data were analyzed using descriptive statistics,

and other insects on Glycine max yields This evaluation was based on the impact of

student’s t-test for the comparison of means of the two

visiting flowers on pollination, the impact of pollination

samples, correlation coefficient (r) for the study of the

on fructification of G. max, and the comparison of

association between two variables, chi-square (χ2) for

yields [fruiting rate, mean number of seeds per pod and

the comparison of two percentages using SPSS statistical

percentage of normal (well developed) seeds] of

software and Microsoft Excel.

treatments 1 and 2. The fruiting rate due to the activity of insects (Fri ) was calculated as follows by Tchuenguem

RESULTS

et al., (2004): Fri = {[(Fr1– Fr2) / Fr1] × 100}

Reproduction system of Glycine max

Where Fr1 and Fr2 are the fruiting rate in treatments 1 and 2.

According to table 2 : the allogamy rate was 6.59% and 5.38% respectively in 2010 and 2011 and

The fruiting rate (Fr) is: Fr = [(F2/F1) × 100]

autogamy rate was 93.41% and 94.62% respectively in

Where F2 is the number of pods formed and F1 the

2010 and 2011. Glycine max (used in our experiments)

number of flowers initially set.

has a mixed reproduction

At maturity, pods were harvested from each

system autogamous -

allogamous, with the predominance of autogamy.

treatment. The mean number of seeds per pod and the

Frequency of A. m. adansonii in the floral entomofauna

percentage of normal seeds were then calculated for

of Glycine max

each treatment. Evaluation

Among the 532 and 422 visits of 24 and 24

the

pollination

efficiency

insect species counted on G. max flower in 2010 and 2011, respectively, A. m. adansonii was the most

Apis mellifera adansonii on Glycine max In 2010, along with the development of

r e p r e s e n t e d insect with 132 visits (24.81 %) and 91

treatment 1 and 2, 11 plants belonging to four subplots

visits (21.56 %), in 2010 and 2011, respectively. The

and carrying 47 flowers were protected using gauze mesh

difference between these two percentages is not

(treatment 3). In 2011 the same experience was repeated

significant (χ2 = 1.39‚ df = 1‚ p > 0.05) (Table 1). In

but on 16 plants carrying 51 flowers. Between 7 and

2010, the highest mean number of A. m. adansonii

9am, of each observation date, the evaluation or the

simultaneously in activity was one per flower (n = 50; s

efficiency pollination of A. m. adansonii on G. max

= 0) and 2.88 per 1000 flowers (n = 60; s = 3.53; maxi

was realized as according of Douka and Tchuenguem

= 19). In 2011, the corresponding figures were one per

(2013). The impact (Frx) of A. m. adansonii to fruiting

flower (n = 50; s = 0) and 1.97 p er 10 0 0 fl ow er s (n

rate was calculated as follows by Tchuenguem et al.,

= 60; s = 2.59; maxi = 12). The difference between the

(2004) the formula:

mean number of foragers per 1000 flowers in 2010 and

Frx = {[(Fr3– Fr2) / Fr3] x 100}

2011 was highly significant (t = 9.19; df = 118, p <

Where Fr3 and Fr2 are the fruiting rates in treatment

0.001).

Activity of Apis mellifera adansonii on Glycine max

(protected

flowers

visited

exclusively

A. m. adansonii) and treatment 2 (protected flowers). The number of seeds per pod, the percentage of normal 1212

Floral reward harvested During each of the two flowering periods, A. m. Journal of Research in Biology (2014) 4(1): 1209-1219

Fohouo and Dounia, 2014 respectively. The difference between the duration of the visit in 2010 and 2011 is higher significant (t = 22.25; df = 221, p < 0.001). For the two cumulated yearsâ&#x20AC;&#x161; the mean duration of a flower visit were 2.55 sec. Foraging speed of Apis mellifera adansonii on Glycine max flowers On the pot of G. max, A. m. adansonii visited between 4 and 24 flowers/min in 2010 and between five and 25 flowers/min in 2011. The mean foraging speed was 11.65 flowers/min (n = 50; s = 5.77) in 2010 and 13.48 flowers/min (n = 50; s = 5.82) in 2011. The Figure 3. Apis mellifera adansonii collecting nectar in a flower of Glycine max

difference between these means is highly significant (t =

adansonii was found to collect nectar intensively and

the mean foraging speed was 12.56 flowers /min.

exclusively (Figure 3).

Effect of climate on foraging activity of Apis mellifera

Relationship between visits and flowering stages

adansonii on Glycine max flowers

- 7.95; df = 98, p < 0.001). For the two cumulated yearsâ&#x20AC;&#x161;

Visits were most numerous when the number of

Climatic condition seem not to influence the

open flowers was highest (Figure 4) Furthermore a

activity of A. m. adansonii. T he correlation was

positive and significant correlation was found between

negative and not significant (r2010 = - 0.34; df = 11; p

the number of G. max opened flowers and the number

> 0.05 and r2011 = 0.28; df = 11; p > 0.05) between the

of A. m. adansonii visits in 2010, as well as 2011

number of A. m. adansonii visits on G. max flowers

(r2010 = 0.90; df = 8; p < 0.05; r2011 = 0.85; df = 8; p <

and the temperature. It was positive and not significant

0.05). Apis mellifera adansonii foraged on G. max

flowers throughout the blooming period, with a peak of

p > 0.05) between the number of A. m. adansonii visits

activity situated between 10 and 11am (Figure 5).

and relative humidity (Figure 6).

Duration of visits per flower

Impact of anthophilous insects on pod formation and

In 2010 and 2011, the mean duration of A. m.

2010 =

0.48; df = 11; p > 0.05 and r2011 = 0.07; df = 11;

seed yields of Glycine max

adansonii visit is 2.50 sec (n = 132; s = 1.34; maxi = 6

During nectar harvest on G. max, foraging

sec) and 2.61 sec (n = 91; s = 1.40; maxi = 6 sec)

insects always shook flowers and are regularly in contact

Figure 4: Variation of number of flowers and number of visits of Apis mellifera adansonii on the flowers of Glycine max in 2010 and 2011. Journal of Research in Biology (2014) 4(1): 1209-1219

Figure 5. Variation of number of flowers and visits of Apis mellifera adansonii on the flowers of Glycine max according to daily time in 2010, 2011. 1213

Fohouo and Dounia, 2014 with the anthers and stigma (Figure 3), increasing cross

normal seeds in opened flowers was higher than that of

pollination possibility of G. max fruiting rate, number of

protected flowers in 2010 and 2011. The percentage of

seeds per pod and percentage of normal seeds in different

the normal seeds due to the action of insects was 24.81%

treatments (Table 2).

in 2010 and 20.90% in 2011. For all the flowers studied,

a - The difference observed

was highly

the percentage of the normal seeds due to flowering

significant between fruiting rate of free opened flowers

insects was 22.85%.

(treatment 1) and that of bagged flowers (treatment 2),

Pollination efficiency of Apis mellifera adansonii on

the first year (χ2 = 248.73, df = 1, p < 0.001) and the

Glycine max

second year (χ2 = 299.84, df = 1, p < 0.001). T he

Apis mellifera adansonii foragers were always

fruiting rate of t r e a t m e n t 1 ( unprotected flowers)

in contact with the stigma and the anthers of G. max

was higher than treatment 2 (protected flowers) in 2010

(contacts with anthers and stigma was 100% for all

and in 2011. The fruiting rate due to the action of insects

visits).

was 5.92 and 5. 81% in 2010 and 2011 respectively.

possibilities of the pollination of G. max flowers.

For the two cumulated years, the fructification rate due to the influence of insects was 5.86%.

C on sequ en t l y

t his

bee

increased

a - the difference observed between the fruiting rate of treatments 2 and that of treatment 3 was highly

b - For the mean number of seeds per pod,

significant in 2010 (χ2 = 7.73; df = 1; p < 0.001) as

there was a highly significant difference between

well as 2011 (χ2 = 6.93; df = 1; p < 0.001). T he

treatments 1 and 2 (t2010 = 4315.78; df = 30462; p <

fruiting rate of flowers exclusively visited by A. m.

0.001; t2011 = 5958.33; df = 30670; p < 0.001).

adansonii (treatment 3) was higher than those of bagged

Consequently, a high mean number of seeds per pod in

flowers (treatment 2). The fruiting rate due to A. m.

treatment 1 (opened flowers) were noticed compared to

adansonii activity was 14.14% and 11.98% respectively

treatments 2 (bagged flowers). The number of seeds per

in 2010 and 2011. The percentage of the fruiting rate

pod attributed to the activity of insects was 26.11% in

due to A. m. adansonii activity was 13.06 %

2010 and 36.47% in 2011, giving an overall mean of 31.29%.

b - There was a highly significant difference between treatments 2 and 3 (t = 64.76; df = 14821; p <

c - There was a highly significant difference

0.001) the first year and the second year (t = 49.28; df =

between the percentage of normal seed of treatment 1

14023; p < 0.001). High mean number of seeds per pod

and that of treatment 2 in the first year (χ2 = 4329.98; df

of flowers of treatment 3 was noticed compared to

= 1; p < 0.0001) as well as the second year (χ2 =

flowers of treatment 2. The augmentation of the number

6094.38; df = 1; p <0.0001). Thus, the percentage of

of seeds per pod due to A. m. adansonii was 36.95% and B

Figure 6. Daily distribution of A. m. adansonii visits on 17187 and 17866 G. max flowers over 10 days in 2010 (A) and 10 days in 2011 (B) respectively, mean temperature and mean humidity of the study site. 1214

Journal of Research in Biology (2014) 4(1): 1209-1219

Fohouo and Dounia, 2014 Table 1. Diversity of floral insects on Glycine max in 2010 and 2011, number and percentage of visits of different insects Insects Order Hymenoptera

Family Apidae

Halictidae Megachilidae

Formicidae Vespidae Sphecidae Lepidoptera

Diptera

Hemiptera Orthroptera Nevroptera Total

Pieridae

Nymphalidae Acraeidae Muscidae Drosophilidae Syrphidae Calliphoridae Coreidae Pyrrhocoridae

2010 Genus, species, sub-species

Apis mellifera adansonii Amegilla sp. 1 n Xylocopa sp. 1 n Macronomia vulpina n Lipotriches collaris n Chalicodoma sp.1 n Megachile sp. 1 n Megachile sp. 2 n Polyrachis sp. 1 sh Synagris cornuta n (1 sp.) n Philanthus triangulum pr (1 sp.) pr Catopsilia florella n (sp. 1) n (sp. 2) n (1 sp.) n Acraea acerata n Musca domestica n Drosophila sp. 1 n (1 sp.) n (1.sp.) n Anoplocnemis curvipes n Dysdercus voelkeri n (sp.1) lv (sp.2) lv (sp.1) pr (sp.2) pr 28 species

2011

p1%

p2%

132 4 3 87 56 13 3 0 79 11 1 6 1 28 17 12 19 13 26 12 2 3 1 1 0 0 2 0

24.81 0.75 0.56 16.35 10.53 2.44 0.56 0 14.85 2.07 0.19 1.13 0.19 5.26 3.20 2.26 3.57 2.44 4.89 2.26 0.38 0.56 0.19 0.19 0 0 0.38 0

91 0 1 51 49 2 1 4 62 4 0 2 0 29 8 3 23 17 49 8 3 0 1 2 5 2 1 4

21.56 0 0.24 12.09 11.61 0.47 0.24 0.95 14,69 0.95 0 0.47 0 6.87 1.90 0.71 5.45 4.03 11.61 1.90 0.71 0 0.24 0.47 1.18 0.47 0.24 0.95

532

100

422

100

Comparison of percentages of Apis mellifera adansonii visits for two years: Ď&#x2021;2 = 1.39 ([df = 1; P > 0.05]). n1: number of visits on 17187 flowers in 10 days. n2: number of visits on 17866 flowers in 10 days. p1 and p2: percentages of visits. p1 = (n1 / 532) x 100. p2= (n2 / 422) x 100. n: Visitor collected nectar. lv: Visitor eating leaves. sh: visitor shelter pr: Predation. sp.: Undetermined species.

Journal of Research in Biology (2014) 4(1): 1209-1219

1215

Fohouo and Dounia, 2014 35.65% respectively in 2010 in 2011. The percentage of

attributed to the variation of the number of colonies of

the mean number of seeds per pod attributed to the

this honey bee around the experimental site. The peak of

activity of A. m. adansonii was 36.30%.

activity of A. m. adansonii on G. max flowers was at

c - There was highly significant difference

between 10 and 11am, which correlated to the period

between the percentage of normal seed of treatment 3

of highest availability of nectar on G. max flowers. The

and that of treatment 2 in first year (χ2 = 67.76; df = 1;

positive and highly significant correlation between the

p < 0.001) as well as the second year (χ2 = 58.58; df

number of G. max flowers and the number of A. m.

= 1; p < 0.001). The percentage of normal seeds in

adansonii visits indicates the attractiveness of G. max

treatment 3

nectar

was higher than in treatment 2. The

with respect

this bee. The significant

percentage of the normal seeds due to A. m. adansonii

difference observed between the duration of visits in

was 32.61% in 2010 and 29.26% in 2011. T he

2010 and 2011 could be attributed to the availability of

percentage of the number of seeds per pod attributed to

nectar, the floral morphology of this crop or the variation

the activity of A. m. adansonii was 30.93%.

in the diversity of flowering insects from one year to another. At Maroua in 2010 and 2011 (in the rainy season),

DISCUSSION Honey bee was the main floral visitor of

harvested

m. adansonii intensely and regularly

nectar on the flowers of G. max during

G. max during the observation period. This bee has

flowering periods. This could be attributed to the needs

been reported as the main floral visitor of this Fabaceae

of colonies during the flowering period. During our

in USA (Rortais et al., 2005) and Brazil (Milfont et al.,

investigations, the interruption of visits by other insects

2013). Apis mellifera adansonii was also shown to be the

or the same honey bee reduced the duration of A. m.

most abundant floral visitors of other Fabaceae members

adansonii

such as Phaseolus coccineus in Yaoundé, Cameroon

Cameroun by Tchuenguem et al., (2009b) and Douka

(Pando et al., 2011a), and Phaseolus vulgaris in

and Tchuenguem (2013) on flowers of Vigna

Ngaoundéré, Cameroon (Kingha et al., 2012) and in

unguiculata (L.) (Fabaceae) and Phaseolus vulgaris

Maroua by Douka and Tchuenguem (2013). The

(Fabaceae)

significant difference between the percentages of A. m.

adansonii visits for the two studied years could be

pollination of G. max flowers. During the collection of

visits.

Similar

results

respectively.

were

found

indicates

that

adansonii can increased the possibility of

Table 2. Glycine max yields under pollination treatments. Treatment

Seeds / Pod

Total seeds

Normal % normal seed seed

Year

Flowers

Pods

Fruiting rate

Unlimited visits

2010

17187

15688

91.28%

Mean 3.14

sd 1.42

48853

42609

87.22

Protected plot

2010

17208

14776

85.87%

2.32

1.01

34162

22415

65.61

Protected plot

2011

17866

16697

93.46%

3.92

2.06

66020

57137

86.54

Bagged flowers

2011

15875

13974

88.03%

2.49

1.52

63176

43250

68.46

A. m. adansonii

2010

100.00%

3.68

1.84

152

148

97.37

A. m. adansonii

2011

100.00%

3.87

1.88

217

201

92.63

1216

Journal of Research in Biology (2014) 4(1): 1209-1219

Fohouo and Dounia, 2014 nectar, A. m. adansonii foragers regularly come into

differ between plant varieties and /or region.

contact with the stigma and carry the pollen to the anthers for stigma. The weight of A. m. adansonii sh oot t h e

CONCLUSION

fl ower s of G. max dur in g nectar collection and this

This study reveals that t h e v a r i e t y o f G. max

movement played a positive role in liberation of pollen

st udi ed is a nectariferous bee plant that obtained

by anthers for the optimal occupation of the stigma.

benefits from the pollination by insects among which A.

This phenomenon was also reported by Ahrent and

m. adansonii is the must important. The comparison of

Caviness (1994) and Rortais et al., (2005) on G. max.

pods and seeds set of unprotected flowers with that of

Thus in addition to their direct pollination role,

flowers visited exclusively

A. m. adansonii foragers also indirectly effected self-

underscores the value of this bee in increasing pods and

pollination and cross-pollination of G. max flowers. The

seed yields as well as seed quality. The installation of

positive and significant contribution of A. m. adansonii

A. m. adansonii c o l o n i e s to G. max fi el d s should

in pods, seed yields and percentage of normal seeds of

be recommended for the increase of pod and seeds

G. max is justified by the action of this bee on

yields of this valuable crop.

adansonii

pollination. The similar have been obtain in Britain (Kendall and Smith, 1976) on Phaseolus coccineus

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(Fabaceae), USA (Ibarra-Perez

1999) on

Ahrent DK, Caviness CE. 1994. Natural cross-

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al.,

(Fabaceae),

(Cameroon) (Kingha et al., 2012) on Phaseolus vulgaris (Fabaceae),

Maroua

(Cameroon)

(Douka

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absence

Crop. Sci. 34 (2): 376-378. Borror DJ, White RE. 1991. Les insectes de l’Amérique du Nord (au nord du Mexique). Broquet (ed.), Laprairie. p. 408.

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Boyeldieu J. 1991. Produire des grains oléagineux et

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protéagineux. Ed. Tec et Doc/Lavoisier Col. Agriculture

treatment with unlimited visits or treatment visiting

d’aujourd’hui. 115 - 152

exclusively by A. m. adansonii compared to treatment with protected, showing that insect visits were effective in increasing cross-pollination or self- pollination. Our results confirmed those of Caviness (1970), Ahrent

Caviness CE. 1970. Cross-pollination in the soybean. In : The indispensable pollinators, Ark. Agr. Ext. Serv. Misc. Pub. 127: 33-36.

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Demarly Y. 1977. Genetic and amelioration of plants.

et al., (2013) who revealed that G. max flowers set little

Masson, Paris. p. 577.

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insects

was

not

always

needed.

Woodworth (1922) showed that self-pollination of G. max flowers produced as many pods and seeds as

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Apidae)

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vulgaris

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Phaseolus

multiflorus and Phaseolus vulgaris by honeybees.

(Ngaoundere-Cameroon) J. Agric. Extension and Rural

vulgaris

(Fabaceae)

flowers

Dang

Apicult. Res. 5: 87-91.

Development. 4 (10): 330-339.

Free JB. 1993. Insect pollination of crops. Academic

Letouzey

Press. p. 544

phytogéographique du Cameroun au 1/500000. Inst.

Freitas BM. 1997. Number and distribution of cashew (Anacardium occidentale) pollen grains on the

1985.

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carte

Carto. Intern. Végétation, Toulouse et Inst. Rech. Agron., Yaoundé.

bodies of its pollinators, Apis mellifera and Centris

Milfont MO, Rocha EEM, Lima AON, Freitas BM.

tarsata. J. Apicul. Res. 36 (1): 15-22.

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Gallais A, Bannerot H. 1992. Amélioration des espèces végétales cultivées. INRA, Paris.p.768.

wild pollinators, a sustainable alternative to pesticides and autopollination. Environ. Chem. Lett. 11(4) :335341. Doi 10.1007/s10311-013-0312 .8.

Hymowitz T. 1970. On the domestication of the soybean. Econ. Bot. 24 (4): 408-421.

Minader. 2010. Annuaire des Statistiques du Secteur Agricole, Campagnes 2007 & 2008. Direction des

Hymowitz T, Harlan JR. 1983. Introduction of soybean

Enquêtes et des Statistiques, Agricoles. AGRI-STAT

to North America by Samuel Bowen in 1765. Econ. Bot.

Cameroun n° 16, p.98.

37 (4): 371-379.

Pando JB, Tchuenguem FF-N, Tamesse JL. 2011a.

Ibarra-Perez FJ, Barnhart D, Ehdaie B, Knio KM,

Foraging and pollination behaviour of Xylocopa calens

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coccineus

Jacob-Remacle A. 1989. Comportement de butinage de

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l’abeille domestique et des abeilles sauvages dans des

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vergers de pommiers en Belgique. Apidologie. 20 (4):

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271-285.

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Kendal DA, Smith BD. 1976. The pollinating efficiency of honey bee and Bumblebee visits to flowers of the

and nectar consumed by different categories of bees. Apidologie., 36 (1): 71-83).

runner bean Phaseolus coccineus L. J. Applied Ecology

Tchuenguem FF-N, Messi J, Brückner D, Bouba B,

13 (3): 749-752.

Mbofung G, Hentchoya HJ. 2004. Foraging and

Kuete M, Melingui A, Mounkam J, Nofiele D. 1993. Nouvelle géographie du Cameroun. EDICEF, Paris. p. 207.

Brückner D. 2012. Foraging and pollination activities

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mellifera adansonii) on Callistemon rigidus flowers in Ngaoundéré

(Cameroon).

Cam.

Acad.

Sci.

4(2): 133-140.

Kingha BMT, Tchuenguem FF-N, Ngakou A, of Xylocopa olivacea (Hymenoptera, Apidae)

pollination behaviour of the African honey bee (Apis

Tchuenguem FF-N, Ngakou A, Kengni BS. 2009b. Pollination and yield responses of cowpea (Vigna unguiculata L. Walp.) to the foraging activity of Apis Journal of Research in Biology (2014) 4(1): 1209-1219

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adansonii

Ngaoundéré

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(Cameroon).

Afr.

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8 (9): 1988-1996. Tien HH, Hien TM, Son MT, Herridge D. 2002. Rhizobial Inoculation and N2 fixation of soybean and mungbean in the Eastern region of South Vietnam. In” inoculants and nitrogen Fixation of legumes in Vietnam”. Edited by D. Herridge. ACIAR proceedings 109e. USDA. 2002. Oilseeds : World markets and trade. Foreign Agricultural Service. Circular series FOP. Woodworth CM. 1922. The extent of natural crosspollination in soybeans. Jour. Amer. Soc. Agron. 14: 278-283.

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Journal of Research in Biology (2014) 4(1): 1209-1219

1219

Journal of Research in Biology

An International Scientific Research Journal

Original Research

Journal of Research in Biology

Determining the Natural Gypsophila L. (Coven) Taxa Growing in Tunceli (Turkey) Authors: Mustafa Korkmaz1* and Hasan Ozçelik2.

ABSTRACT: 56 species belonging to 60 taxa (out of 126 species in the World) of Caryophyllaceae family grows naturally in Turkey with Gypsophila sps L. as the third largest genus. The endemism ratio of the genus is 60% in Turkey. Because Turkey is the gene center of Gypsophila and economically very valuable; determining the geographic distribution and biological characteristics of the taxa is very necessary. They have well-developed roots, that prevent soil erosion. Because of containing saponin (10-25 %) in their root, its extract is used as fire extinguisher, gold polisher, Institution: cleaner and softener of delicate fabrics and crispness giving substance for halva. It is 1. Erzincan Üniversity, also used for making liqueur, herbal cheese, ice cream and some other foods. Some Science and Arts Faculty, taxa are boron hyper acumulators and vegetative mining can be conducted by hyper Department of Biology, accumulation. They are also thought to be the cleaning tools for toxid areas by Erzincan-Turkey. fitoremediation. In this study, 12 records from eight Gypsophila taxa were collected around 2. Süleyman Demirel Tunceli. These are G. aucheri Boiss. (1), G. elegans Bieb. (1), G. pallida Stapf. (2), Üniversity, Science and Arts G. perfoliata L. var. perfoliata (1), G. ruscifolia Boiss. (3), G. sphaerocephala Fenzl ex Faculty, Department of Tchihat var. cappadocica Boiss. (1), G. venusta Fenzl (1) and G. viscosa Murray (2). Biology, Isparta-Turkey. With addition of G. briquetiana Schischk. and G. hispida Boiss. the total number is reaching to 10 and it shows that the city is an important diversity center of the genus. G. aucheri, G. briquetiana and G. sphaerocephala var. cappadocica are endemic to Turkey and G. pallida, G. perfoliata L. var. perfoliata, G. venusta and G. viscosa are determined to be new records for Tunceli. Corresponding author: Mustafa Korkmaz.

Keywords: Coven, Gypsophila, Habitat, Biodiversity, Tunceli, Turkey.

Email Id:

Article Citation: Mustafa Korkmaz and Hasan Ozçelik. Determining the Natural Gypsophila L. (Coven) Taxa Growing in Tunceli (Turkey). Journal of Research in Biology (2014) 4(1): 1220-1227

Web Address:

Dates: Received: 04 Feb 2014

http://jresearchbiology.com/ documents/RA0421.pdf.

Accepted: 05 Mar 2014

Published: 16 April 2014

1220-1227 | JRB | 2014 | Vol 4 No 1

www.jresearchbiology.com

Korkmaz and Ozçelik, 2014 2008; Korkmaz et al., 2010; Korkmaz and Özçelik,

INTRODUCTION Caryophyllaceae family distributes mostly in

2011a).

Mediterranean region of southern hemisphere. It has a

Turkish Covens are commonly obtained from

large diversity with over 2000 species. Gypsophila L.

Gypsophila graminifolia Bark. G. arrostii Guss.var.

genus, which has 126 species on the World, has natural

nebulosa (Boiss. and Heldr.) Bark., G. eriocalyx Boiss.,

distribution in the Irano-Turanian and Mediterranean

G. bicolor

phytogeographic regions (Williams, 1989; Sumaira et al.,

G. venusta Fenzl

2008). There are about 500 species of Caryophyllaceae

gypsophiloides Fenzl. (İnan, 2006; Kılıç, et al., 2008).

family in Turkey. More than half of totally 126

G. ruscifolia Boiss. and G. bitlisensis Bark. are the least

Gypsophila species in the world are found in Caucasian,

preferred species. The most preferred species are

the North Iraq and the North Iran regions. There are

G. bicolor, G. arrostii and A. Gypsophiloides (Baytop,

about 56 Gypsophila species found in Turkey. Many of

1984; Özçelik, and, Özgökçe, 1999; Korkmaz and

them are known from the type collection. G. heteropoda

Özçelik, 2011a).

(Freyn&Sint.) Grossh., G. perfoliata L., subsp. venusta and Ankyropetalum

Freyn & Sint. subsp. minutiflora Bark. is a rare endemic

Saponin chemical was first produced from the

taxon peculiar to Cappadocica sub region in Inner

roots of Saponaria officinalis (Baytop, 1984). The

Anatolia of Turkey and an endangered taxa on global

amount of saponin in the roots of Gypsophila taxa differs

scale (Ekim et al., 2000; Ozhatay et al., 2005).

from 4 % to 25 % (Sezik, 1982). Gypsophila bicolor

Gypsophila

(Van Çöveni), G. arrostii var. Nebulosa (Beyşehir,

the

third

biggest

genus

Caryophyllaceae family after Silene L. and Dianthus L.

Isparta

Çöveni),

(Davis, 1967; Davis et al., 1988; Güner et al., 2000;

Çelik et al., 2008; Korkmaz and Özçelik, 2011b).The

(Çorum-Yozgat Çöveni) are most preferred taxa for

most important factor for the distribution of this genus is

obtaining coven extract in Turkey (H´eroldand Henry,

the soil structure which contains gypsum, lime and

2001; Battal, 2002).

venusta

subsp.

perfoliata Venusta

(Niğde

and

Çöveni), eriocalyx

calcium; these are important for these plants to grow.

Soap root extract is composed of sugar, resin and

There are gypsum habitats around Sivas, Çankırı,

saponin. It protects the plant from germ and fungal

Çorum, Ankara, Eskişehir, Niğde and Erzincan. Because

infection, increases the nutritive value and facilitates the

of that, Gypsophila taxa are rich in these areas.

digestion. The production phases of the extract starts

Soap root has been exported from Anatolia for a

with cutting the roots in the form of chips and continuous

long time. The collection of coven from natural habitats

with boiling them for two times. After second boiling

and extraction have been increasing rapidly especially in

stage the extract can be obtained. (Korkmaz et al., 2010;

the Eastern and South-east Anatolia for nearly 40 years

Korkmaz and Özçelik, 2011a).

(Kılıç, et al., 2008). In Turkey Gypsophila taxa are

The main areas of the use of them are in the food

generally known by the name “Çöven Otu” and they are

industry, the chemistry, in hygiene industry, in

mostly used by the public for different purposes. The

horticulture, in mining, in whitening gold and in fire

word “Soaproot” or “Soapworth” terms are used for

extinguishers. They have antimicrobial effect and used in

Gypsophila species; in Europe the members of the genus

medicines. Every year the average export of soap root

are widely known as “Baby’s Breath”. In Turkey the

from Turkey is about 90 tones by gaining approximately

plants are also called “Dişi Çöven, Tarla Çöveni, Helva

66 000 US Dollars (Baytop, 1984; Korkmaz and Özçelik,

Çöveni, Şark Çöveni” by the local people (Kılıç, et al.,

2011a; Özçelik and Özgökçe, 1996).

1221

Journal of Research in Biology (2014) 4(1): 1220-1227

Korkmaz and Ozçelik, 2014 This study was aimed to determine the Gypsophila taxa

herbarium specimen. Economic importance of the taxa is

naturally distribute in the province of Tunceli city of

given according to our early papers (Özçelik and

Turkey.

Özgökçe, 1999; Korkmaz et al., 2010; Korkmaz and Özçelik, 2011a,b).

MATERIALS AND METHODS

As it is given in the Table-2, endemic taxa and

Material of this study contains Gypsophila taxa

the risk categories, phytogeographic regions, altitudes,

growing around Tunceli. With regard to this aim we have

life forms and new records have been determined.

collected eight taxa of the genus from 13 different

Turkish names of Gypsophila taxa grows around Tunceli

localities in the area. Collection date, record number,

have been determined from Türkiye Bitkileri Listesi

habitat types and some other properties of the identified

(Güner et al., 2012) as they were given in Table 2.

taxa were determined (and given in Table 1). For the

Endemic taxa of the genus and their threat categories

identification of taxa Flora of Turkey and the East

have been determined from Ekim et al. (2000) and given

Aegean Islands (Davis, 1967) has been used extensively.

in the same table.

Identifications were done with the help of stereo-zoom microscope. Identified samples were converted to Table 1. Locality and habitat informationof Gypsophila taxa collected around Tunceli Taxon

Record number

Date

Locality

G. aucheri Boiss.

K: 1769

03.07.2009

G. elegans Bieb.

K: 1741

02.07.2009

K: 1740

02.07.2009

K: 1748

02.07.2009

K: 1745

02.07.2009

K: 1746

02.07.2009

K: 1760

02.07.2009

K: 1761

02.07.2009

G. sphaerocephala Fenzl ex Tchihat var. cappadocica Boiss.

K: 2588

12.06.2011

K: 2638

11.07.2011

G. venusta Fenzl

K: 1749

02.07.2009

G. viscosa Murray

K: 1750

02.07.2009

K: 1752

02.07.2009

Tunceli: Tunceli-Pertek, 10 km to Pertek Tunceli: Erzincan- Pülümür, near to Pülümür Tunceli: Erzincan- Pülümür, near to Pülümür Tunceli: Tunceli- Ovacık, 40 km to Ovacık Tunceli: Pülümür-Tunceli, near to Pülümür Tunceli: Pülümür-Tunceli, 30 km to Tunceli Tunceli: Tunceli-Ovacık, 10 km to Ovacık Tunceli: Ovacık, Munzur Çayı Gözeleri Tunceli-Erzincan, Munzur Mountain Tunceli-Erzincan Munzur Mountain Tunceli: Tunceli- Ovacık, 25 km to Ovacık Tunceli: Tunceli Ovacıkarası, 25 km to Ovacık Tunceli: Tunceli-Ovacık, 10 km to Ovacık

G. pallida Stapf.

G. perfoliata L. var. perfoliata

G. ruscifolia Boiss.

Habitat Rocky places Rocky places Rocky places Inclined slopes Rocky slopes Rocky slopes Flowing slopes Rocky places Rocky slopes Slopes Rocky slopes Rocky slopes Rocky places

K: Korkmaz Journal of Research in Biology (2014) 4(1): 1220-1227

1222

Korkmaz and Ozçelik, 2014 Table 2. Taxonomic information of Gypsophila taxa growing around Tunceli No

Taxon name (Turkish name)

Endemic

Fl.

P.G. region

Altitude (m)

Life form

New record or recorded before

G. aucheri Boiss. (Taş Çöveni)

Endemic (VU)

6-7

Ir.-Tur.

1200-1600

Tunceli, Pertek

*G. briquetiana Schischk. (Gül Çevgeni)

Endemic (LR)

7-8

Ir.-Tur.

1700-2500

Tunceli, Ovacık, Munzur Mountain

G. elegans Bieb. (Hoş Çöven)

6-7

Ir.-Tur.

650-2600

New record to Tunceli

*G. hispida Boiss. (Kıllı Çöven)

6-7

Ir.-Tur.

1100-2150

Tunceli, between Tunceli and Ovacık

G. pallida Stapf. (Şark Çöveni)

6-8

Ir.-Tur.

850-2000

New record to Tunceli

G. perfoliata L. var. Perfoliata (Helvacı Çöveni)

6-8

1000-1500

New record to Tunceli

G. ruscifolia Boiss. (Acem Çöveni)

6-7

Ir.-Tur.

300-1800

Tunceli, Ovacık

G. sphaerocephala Fenzl ex Tchihat var. cappadocica Boiss.

Endemic (LR)

7-8

Ir.-Tur.

800-1900

Tunceli, Munzur Mountain

G. venusta Fenzl (Kara Çöven)

5-7

Ir.-Tur.

300-1600

New record to Tunceli

G. viscosa Murray (Sadırlı Çöven)

4-6

Ir.-Tur.

350-1400

New record to Tunceli

* :Gypsophila taxa not available in the area, P: Perennial, A: Annual, P.G.: Phyto-geographic, Fl.: Flowering period RESULTS AND DISCUSSION

G. aucheri, G. briquetianaand G. sphaerocephala var.

The results of the study are summarized in Table

cappadocica are endemic taxa available in the vicinity.

-1 and Table-2. As seen in Table-1, 8 Gypsophila taxa

Threat (risk) category of G. aucheri is Vulnerable (VU)

were collected from the area in 2009 and 2011. All of the

and the other two taxa is at the category of Low Risk

plant samples were collected from Pülümür, Tunceli,

(LR). Flowering periods of the taxa changes from April

Ovacık and Munzur Mountains. Generally, the collected

to August. All of the determined taxa are Irano-Turanian

plants are naturally grown in rocky and slopy places.

phytogeographic region elements and distributes from

Photograph of all collections were taken during the field

800 to 2500 m altitudes in the area.

work. Totally 8 Gypsophila taxa were collected from 13

and G. viscose are annual life forms and the others are

different localities. As seen in Table-2 there are 10

perennial life forms. G. aucheri, G. briquetiana,

Gypsophila taxa determined in the flora of Tunceli.

1223

elegans,

hispida,

G. elegans

ruscifolia

and

Journal of Research in Biology (2014) 4(1): 1220-1227

Korkmaz and Ozçelik, 2014 G. sphaerocephala var. cappadocica are early recorded

as forming a natural border between Erzincan and

in Tunceli but, G. pallida, G. perfoliata var. perfoliata,

Tunceli. The width of the mountain is 25-30 km and the

G. venusta and G. viscose (4 taxa) are new records.

length of it is 100-130 km. Altitude of the area changes

Habitat types of Gypsophila taxa growing naturally in

from nearly 850 to 3462 m. The climate of the area is hot

the province are rocky places, in clined or flowing slopes

and dry summers and long and snowy winters.

and slopes of mountains. Their flowering period starts in

According to the study there are 1407 vascular plant

July. The general vegetation type of the plants are arid or

species. The number of endemic species is 275 and some

semiarid steppes.

of them were described as new to science. In this study

Soap roots have economic value in medicine,

briquetiana

Schischk.,

sphaerocephala,

food, decoration and cleaning and chemistry to produce

G. ruscifolia, G. elegans Bieb, G. bitlisensis Bark. and

saponin. It is used as fire extinguisher, gold polisher,

G. hispida Boiss. are given in the list of the plants.

fabric, cleaner and for purification of contaminated soil

Munzur Dağları is one of the most important ÖBA

such as by removing the boron. In addition, it is possible

(Önemli Bitki Alanı) of Turkey with its very rich floristic

boron

diversity. Munzur Valley is also an important national

hyper-acumulation from soil to the upper parts of the

park of the country. There are 43 plant species peculiarto

plant (Babaoğlu et al., 2004; Korkmaz and Özçelik,

Munzur Dağları. In addition to the study of Yıldırımlı

2011a). Turkish soaproot is mostly obtained from

(1995) Özhatay et al. (2005), this is another important

G. graminifolia, G. bicolor, G. arrostii var. nebulosa,

study on biological diversity of the mountains.

G. eriocalyx, G. perfoliata var. anatolica, G. venusta and

Gypsophila briquetiana Schischk., Gypsophila elegans

Ankyropetalum gypsophiloides species and the gene

Bieb. and Gypsophila ruscifolia Boiss. are three species

center of both of the species is Turkey (Korkmaz and

of the genus growing in the area of Munzur mountains

Özçelik, 2011a,b). The harvest time of these plants is

(Koyuncu and Arslan, 2009). Polat et al. (2012)

from March to June. Because the roots of these plants are

evaluated ethno botanical studies performed in the

generally used, the plants don’t produce seeds for the

Eastern Anatolian region including Tunceli. According

next years. So, the plants are increasingly disappearing

to this study there are only five ethnobotanical study

from the nature and under the threat of extinction. This

(Tuzlacı ve Doğan, 2010; Yıldırımlı, 1985; 1991; 1994

problem becomes more important when the plants are

a;b) conducted in Tunceli. Also in another study

rare or endemic. Because of unemployment soap roots

performed by Karlıdağ in (2009) related with both of

have been collected for a long time in the rural parts of

Elazığ and Tunceli, they determined local names and

the country. For preservation of Gypsophila species they

medicinal uses of 53 plants.The least studied cities in

should not only be collected from nature but also its

East Anatolian region are Ağrı, Ardahan, Bingöl, Bitlis,

cultivation should be planned and other soap root

Erzincan, Kars, Muş, Hakkari and Tunceli. So, it is

yielding plant species should be identified.

necessary to record and prevent ethnobotanical culture in

perform

vegetative

mining

The most important floristic study related with Tunceli in the area is Flora of Munzur Dağları

these cities by conducting news tudies (Polat et al., 2012).

(Yıldırımlı, 1995). The mountains are situated between Erzincan and Tunceli in B7 grid square and in

CONCLUSION:

Irano-Turanian phytogeographic region. It starts from

There are 60 naturally growing Gypsophila taxa

Kemaliye and reach to Pülümür at the west-east direction

in the Turkey. Many species of the genus are highly

Journal of Research in Biology (2014) 4(1): 1220-1227

1224

Korkmaz and Ozçelik, 2014 potential to be used in economy. G. sphaerocephala and

the support provided by the institution

G. perfoliata are known as boron hyper accumulators and they are very important for boron mining. Because

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Journal of Research in Biology (2014) 4(1): 1220-1227

Journal of Research in Biology

An International Scientific Research Journal

Original Research

Journal of Research in Biology

Distribution pattern of birds in Banni Grassland of Kachchh district, Gujarat, India Authors: ABSTRACT: Mukesh H. Koladiya1, ArunKumar Roy Mahato2, Birds are interesting group of animals which are distributed in all major types Nikunj B. Gajera3 and habitat. Banni is one of the large grassland of India invaded by Prosopis juliflora, an Yatin S. Patel4. alien plant species. Invasion of this species and some other natural and anthropogenic factor leads the grassland converted into a mixture of heterogeneous habitats. A study was attempted to understand the distribution of birds in this heterogeneous grassland. The habitats were identified based on dominant species of plants. The population estimates of birds were surveyed using line transects method and point Institution: count census method. 1,2,3. Gujarat Institute of A total of 91 species were recorded during the survey in the various habitats Desert Ecology, Bhuj, of this grassland. Among the seven habitats, sparse Prosopis was the most diverse Kachchh-370001, Gujarat. habitat for bird species whereas Prosopis-Capparis was the least diverse habitat for bird species. The highest mean population density of birds were recorded in Prosopis4. Samarth Organization Capparis (15.9 individuals/km2), while lowest recorded in sparse Prosopis habitat (9 Trust, Vijapur, Mehsanaindividuals/km2). It was found that, Prosopis-Salvadora (23.10Âą9.47) was the most 382870, Gujarat. dense and Prosopis-Capperis (8.84Âą5.26) was the least dense habitat for common birds of Banni grassland. In conclusion, bird species diversity and their population density estimates were varied among the various heterogeneous habitats of Banni grassland both in time and space gradients. Corresponding author: Mukesh H. Koladiya.

Keywords: Bird, distribution, density, habitat, Banni grassland, Kachchh

Email Id:

Article Citation: Mukesh H. Koladiya, ArunKumar Roy Mahato, Nikunj B. Gajera and Yatin S. Patel. Distribution pattern of birds in Banni Grassland of Kachchh district, Gujarat, India. Journal of Research in Biology (2014) 4(1):1228-1239

Web Address:

Dates: Received: 10 Feb 2014

http://jresearchbiology.com/ documents/RA0422.pdf.

Journal of Research in Biology An International Scientific Research Journal

Accepted: 24 Feb 2014

Published: 16 April 2014

1228-1239 | JRB | 2014 | Vol 4 | No 1

www.jresearchbiology.com

Koladiya et al., 2014 understanding on the distribution pattern and habitat

INTRODUCTION: Various group of animals varied from survival

preference of bird communities over heterogeneous

strategies in a landscape which are evolved in long

environment is very much essential for conservation and

course of evolution. The distribution patterns of animals

management of birds in regional as well as in local

in various habitats are preferred in response to various

environment (Kattan and Franco, 2004).

uses and selective processes (Clark and Shutler, 1999).

Banni grassland is one of the rich areas of birds

The distributions of life forms are not typically random

due to its varied micro-habitat and act as a seasonal

in the habitat and it is generally assumed that non-

wetland. The distribution pattern of birds across the

random distribution of life forms is due to natural

grassland is very less understood due to the lack of study

selection (Southwood, 1977).

The distribution range

in the area. Therefore, the present study was conducted

across a heterogeneous landscape may depend on the

to understand the pattern of distribution of birds in time

habitat selected by the species, and animal which favors

and space gradient in the grassland for their conservation

their distribution in a greater proportion of the habitat

and management.

(Veech et al., 2011). Banni grassland is one of the largest remnant grassland of India. The landscape of this grassland is flat and most part of it is filled with water during monsoon which makes the grassland as a wetland.

MATERIALS AND METHODS: Study Area: Banni, the largest remnant grassland in India,

The soil

situated on the south-west portion of the Kachchh

salinity is normally high in most of the part due to its

Biosphere Reserve (KBR) and attached to the fringes of

connection with Great Rann of Kachchh (GRK), a salt

greater Runn of Kachchh (23°19' to 23°52' N latitude and

inflated marshy land. To protect the grassland from salt

68°56' to 70°32' E longitude), encompassing an area of

intrusion from GRK, Prosopis juliflora was introduced

over 2,600 km2 is taken into consideration for our study

in fringe areas of GRK to check desertification in Banni

(Fig-1). A large tract of the southern part of Banni

grasslands. In present, P. juliflora is proved to be an

grassland is marshy land and salty waste remains a

invasive species for the grassland and now major part of

wetland in the monsoon season, known as Little Rann of

the grassland is invaded by the species.

Banni, which separates the Banni grassland from the

Birds are very important animal for this

mainland of Kachchh district (Shah and Somusundaram,

ecosystem as they are good indicators of biodiversity.

2010). The climate of the Banni is arid and semi-arid

Birds are one of the typical groups of animal distributed

type therefore, the temperature is high during most of the

in large landscape and even some species prefer to live in

time and it reaches a maximum of 48°-49°C during May-

heterogeneous environment distributed over continents.

June and low during winter season (8°-10°C) in the

To understand the processes of habitat selection and

month of January and February. The average yearly

preference by birds is dependent on an accurate

rainfall of this grassland is 317 mm with scanty rainfall

representation of the patterns of habitat occupancy

and droughts are the common phenomenon of this area.

(Wiens et al., 1987). Birds generally colonize in an area

The grassland is situated in the semi-arid bio-

having presence of suitable habitat for their survival

climatic zone of India. The major part of grassland is

needs (Veech et al., 2011). The distribution pattern of

now invaded by Prosopis juliflora, an invasive alien

birds might also influence by distribution patterns of bird

species. The grassland has varied types of habitat patches

species richness (Shiu and Lee, 2003). The above

that attract large number of birds. Further, the seasonal

1229

Journal of Research in Biology (2014) 4(1): 1228-1239

Koladiya et al., 2014

Figure 1. A map of Banni grassland, and its location in the Kachchh district of Gujarat. water bodies (locally known as Dhandh) inside the Banni region serve as the wintering ground for many migratory species of birds.

Habitat classification: Banni was earlier divided by 10 habitat types by Koladiya et al. (2012). In the present study, the Banni grassland was divided into 7 major habitat types based

METHODOLOGY: A preliminary survey was made to whole of the

on the dominant plant species. Prosopis,

Moderate

Prosopis

It includes; Dense (medium

Prosopis

Banni grassland for identifying transect location and

density), Sparse Prosopis, Prosopis-Capparis Mixed,

number of transect location required for the survey.

Prosopis-Suaeda-Calotropis Mixed, Prosopis-Salvadora

Based on this survey various micro-habitats were

Mixed and Suaeda Dominant. The vegetation of the

identified. A total of 60 km distance was covered by

study area was also recorded by making quadrate on the

walking through various transects. The field data were

line transect and calculated the density of vegetation by

collected by two observers during the whole study period

using Misra (1968).

between the months of June 2009 to May 2011. The

Avi-faunal Survey:

birds were identified using the field guide produced by

The population and distribution of birds were

Ali (1996) and survey was conducted by using standard

recorded using line transect method and point count

data sheet, GPS-Garmin, binocular (8X40) and camera.

census method (Bibby et al., 1992; Bhupathy, 1991). A total of 51 transects were laid down in the whole

Journal of Research in Biology (2014) 4(1): 1228-1239

1230

Koladiya et al., 2014 Table 1. Major plant species density and birds population density in various micro-habitats of Banni grassland Habitat class

Mean individuals of bird/Km2

Vegetation Dominant species

Density/ Ha

Winter

Summer

Monsoon

Dense Prosopis (DP)

Prosopis juliflora

1200.00

12.4

4.50

20.5

Moderate Prosopis (MP)

Prosopis juliflora

833.33

12.3

4.30

17.4

Sparse Prosopis (SP)

Prosopis juliflora

483.33

8.9

2.80

15.3

Prosopis juliflora

733.33 15.5

3.00

29.1

Capparis decidua

1400.00

Prosopis juliflora.

1050.00

Suaeda sps.

2133.30

7.8

4.40

16.6

Calotropis sps.

8933.30 21.4

5.70

17.2

10000.00

13.0

4.20

20.4

13.1±4.50

4.12±0.98

19.5±4.64

Prosopis-Capparis mixed (PC)

Prosopis-Suaeda-Calotropis mixed (PSC)

Prosopis-Salvadora mixed (PS)

Prosopis juliflora

433.33

Salvadora sps.

366.67

Suaeda dominant (SD)

Suaeda sps.

Mean±SD

surveyed area. The presence of individual and group of

and found in all habitats except Suaeda dominant habitat.

birds within 25 m radius of circular plot was made in

The flag ship and dominant species of plants in the seven

every 200 m distance along the line transect. The species

identified habitat were Prosopis juliflora, Capparis

of bird was identified using binoculars and with the help

decidua, Suaeda spp., Calotropis spp. and Salvadora

of Ali and Ripley (1983) and Grimmett et al.(2006).

spp. The density of major plant species calculated in

Generally, the surveys were made during the morning

each habitat type is given in table-1.

(7.30 am to 11.30 am) and afternoon (4.00 pm to 6.30

Species Richness and diversity:

pm) hours of each season during 2009 and 2011.

A total of 91 Species of avi-fauna belonging to

The data recorded during the study was used to

62 genera under 35 families and 11 orders were observed

calculate vegetation density, bird’s population density

during the whole study period (given in Annexure-I).

(Gaston, 1973; Burnham et al., 1980) and tested by

Among the total observed bird species, 59 were resident

ANOVA between micro-habitat using Microsoft Excel

and 32 were migratory in nature. The number of bird

2007.

species recorded in Banni grassland based on their feeding guilds included; granivorous (32 species),

RESULTS AND DISCUSSION:

insectivorous (30 species), frutivorous (12 species),

Habitat category & Vegetation density:

piscivorous (10 species) and others (7 species).

Based

Among the seven identified habitats of Banni

on the transect survey in various seasons, the maximum

grassland Prosopis juliflora is the most dominant species

bird species recorded during monsoon (83 species), next

1231

Journal of Research in Biology (2014) 4(1): 1228-1239

Koladiya et al., 2014

Figure 2. Seasonal Avian species richness in various habitat of Banni grassland to that in winter (67 species) and minimum during

lowest in Prosopis-Capparis mixed (H= 0.91) habitat

summer (32 species).

(fig-2). The above result highlighted that avian species

The total number of avian species was recorded

diversity was also lower in comparison to the species

lower than number of species (163) recorded by Gajera

diversity recorded by Gajera et al. (2012, 2013a, 2013b)

et al. (2012, 2013a, 2013b) in wetland, arid grasslands

in wetland, grassland and mining areas distributed in

and mining areas respectively distributed in western part

western parts of Kachchh district.

of Kachchh district. It is also noted that 56 species of

Distribution of birds in various micro-habitat:

birds recorded alone from the Pena thattah, a seasonal

Out of the total species recorded during the

wetland located in the western part of Banni grassland by

whole study period, the number of bird species recorded

Koladiya et al. (2013).

in 7 identified habitats were as follows; dense Prosopis

The species diversity (Shannon_H) was recorded

(45 species), moderate Prosopis means Prosopis density

to found highest in Sparse Prosopis (H=2.20) habitat and

between more than 500 and less than 1000 individuals/

Figure. 3. Bird species diversity in various habitats of Banni grassland, Kachchh Journal of Research in Biology (2014) 4(1): 1228-1239

1232

Koladiya et al., 2014

Figure 4. Seasonal abundance (%) of birds in Banni grassland of Kachchh, Gujarat ha. (56 species), sparse Prosopis (60 species), Prosopis-

season;

Capparis

Prosopis-Suaeda-

Calotropis are the most preferred habitat during the

Calotropis mixed (50 species), Prosopis-Salvadora

month of summer (Fig-3). The percent of species

mixed (30 species) and Suaeda dominant (40 species)

recorded in each type of habitat in seasonal basis is

respectively. The above result highlighted that sparse

shown in Figure-4.

mixed

(28

species),

moderate

Prosopis

and

Prosopis-Suaeda-

Prosopis was the rich habitat for bird species diversity

We found that the mean population density

and Prosopis-Capparis mixed was the least supportive

(Mean Âą SD) of birds was highest during monsoon

habitat for bird species diversity in Banni grassland. The

season (19.5Âą4.64) and least density during summer

number of species diversity between three season

season (4.12Âą0.98). The seasonal population density of

(summer, monsoon and winter) was significantly varied

birds in various habitats of Banni grassland is given in

(F=14.40, df=2, p<0.001) while species diversity

table-1. It was found that the highest population density

between various habitat were significantly not varied.

of birds was found in Prosopis-Capparis mixed habitat

On analysis of seasonal distribution of bird

(29.1 individuals/km2) during monsoon and least density

species in 7 identified habitats of Banni grassland, it was

was recorded in sparse Prosopis habitat (2.8 individuals/

found that sparse Prosopis, Prosopis-Suaeda-Calotropis

km2)

and dense Prosopis were the preferred habitat during

density of birds recorded in various habitats of Banni

monsoon season; moderate Prosopis, dense Prosopis and

grassland is shown in fig-5. Among the various habitat,

Suaeda dominant are the preferred habitat during winter

the highest mean population density of birds were

1233

during summer season. The mean population

Journal of Research in Biology (2014) 4(1): 1228-1239

Koladiya et al., 2014

Figure 5. Population density of birds in various habitats of Banni grassland, Kachchh

recorded in Prosopis-Capparis (15.9 individuals/km2) 2

the

most

dense

habitat

and

Prosopis-Capparis

and Prosopis-Salvadora habitats (14.8 individuals/km )

(8.84Âą5.26) was the least dense habitat for the common

while lowest mean population density was recorded in

birds of Banni grassland.

sparse Prosopis habitat (9 individuals/km ). The result revealed that the density of birds in Banni grassland was higher in relation to the density of birds recorded by

CONCLUSION: In conclusion, the diversity of birds in banni

Gajera et. al (2013b) in western part of Kachchh.

grassland is rich with sparse Prosopis is the richest

Distribution pattern of common birds:

habitat compare to other habitat in relation to species

We analyse the population density estimates of

diversity. Prosopis juliflora, an invasive alien species of

commonnly occuring 10 species of birds in identified

plant in the grassland is playing major role in the

seven habitat types of Banni grassland (Table-1). It was

distribution of avi-fauna in this region. Prosopis juliflora

found that, Prosopis-Salvadora was the most dense

is the dominant species of plant of this grassland which

habitat of six common species of birds viz. house crow,

provide habitat for nesting of birds and greater visibility

lark, babblar, dove, bee eater and bul bul; sparse

of birds for preying. Based on the results of the study, it

Prosopis was the most dense habitat of pegion and

was found that monsoon season attracts more number of

drongo; dense Prosopis for sand groose and Prosopis-

species of birds in the grassland because large portion of

Suaeda-Capparis was the most dense habitat for

the grassland is converted into seasonal wetland during

francolin. Similarly, Suaeda dominent was the least

the season. However, habitats with dominance of mixed

dense habitat of four species viz. babblar, dove, bee eater

vegetation are the dense in habitat for birds compared to

and bul bul; Prosopis-Capparis and Prosopis-Suaeda-

other habitats of the grassland.

Capparis were the least dense habitat for three species of common birds viz. house crow, francolin, dansgroose

ACKNOWLEDGEMENTS:

and lark, pigeon, drongo respectively. On estimating the

We would like to thank Dr. R. V. Asari, Director,

overall mean density (MeanÂąSD) of common birds, it

Gujarat Institute of Desert Ecology (GUIDE) for

was found that, Prosopis-Salvadora (23.10Âą9.47) was

providing logistic supports and his encouragement. We

Journal of Research in Biology (2014) 4(1): 1228-1239

1234

Koladiya et al., 2014 are thankful to Mr. Yatin Patel for his help in Plant data

(3):166-170.

analysis for the manuscript. We are also thankful to all

Gaston

scientist and scholars of Terrestrial Ecology Division of

populations. J Bomb Nat Hist Soc., 72(2): 271–283.

GUIDE for their help and valuable comments. We are grateful to State Forest Department, Gujarat for providing funds for conducting this study.

AJ.

1973.

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Grimmett R, Inskipp C and Inskipp T. 2006. Pocket Guide to the birds of the Indian sub-continent. Oxford University Press, New Delhi. 384 p.

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2012. Wetland birds of arid region-a study on their

Southwood TRE. 2011. Habitat, the Templet for

diversity and distribution pattern in Kachchh. Columban

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J Life Sci., 13(1&2): 47-51.

Veech JA, Small MF and Baccus JT. 2011. The effect

Gajera NB, Mahato AK Roy and Vijay Kumar V.

of habitat on the range expansion of a native and an

2013a. Status, Distribution and Diversity of Birds in

introduced bird species. J Biog. 38(1): 69–77.

mining environment of Kachchh, Gujarat. Intern

Biod., http://dx.doi.org/10.1155/2013/471618.

Wiens JA, Rotenberry JT. Horne BV. 1987. Habitat Occupancy Patterns of North American Shrubsteppe

Gajera NB, Mahato AK Roy and Vijay Kumar V.

Birds: The Effects of Spatial Scale. Oikos. 48(2):132-

2013b. Status, diversity and conservation of grassland

147.

birds in arid region of Kachchh. Intern J Rec Sci Res., 4 1235

Journal of Research in Biology (2014) 4(1): 1228-1239

Koladiya et al., 2014 Annexure I List of bird species recorded in various habitat of Banni grassland S. No

Family

Scientific Name

Common Name

Habitat

Phasianidae

Francolinus pondicerianus

Grey Francolin

DP, MP, SP, PSC,SD

Upupidae

Upupa epops

Common Hoopoe

MP, SP, SD

Coraciidae

Coracias garrulus

European Roller

MP, SP, SD

Coracias benghalensis

Indian Roller

SP, PC

Merops orientalis

Green Bee-eater

DP, MP, SP, PC, PS

Merops leschenaulti

Chestnut-Headed Bee-Eater

DP, MP, SP, PC, PS

4 5

Meropidae

6 7

Cuculidae

Eudynamys scolopacea

Asian Koel

SP, PC, PS

Centropodidae

Centropus sinensis

Greater Coucal

MP, SP, SD

Psittacidae

Psittacula krameri

Rose-Ringed Parakeet

PC, PS, SD

Apodidae

Apus affinis

House Swift

MP, SP, SD

Strigidae

Bubo bubo

Eurasian Eagle-Owl

DP, MP, SP

Columbidae

Columba livia

Blue Rock Pigeon

DP, MP, SP, PC

Streptopelia decaocto

Eurasian Collared Dove

DP, MP, SP, PC

Streptopelia tranquebarica

Red Collared Dove

DP, MP, SP, PC

Streptopelia chinensis

Spotted Dove

DP, MP, SP, PC

Streptopelia senegalensis

Little Brown Dove

DP, MP, SP

Pterocles exustus

Chestnut-bellied Sandgrouse

DP, MP, SP, PC

Pterocles indicus

Painted Sandgrouse

DP, MP, SP, PC

Circus pygargus

Montagu's Harrier

MP, PSC

Circus aeruginosus

Eurasian Marsh Harrier

DP, MP, PSC

Circus cyaneus

Hen Harrier

DP, MP, PSC

Circus macrourus

Pallid Harrier

DP, MP, PSC

Accipiter badius

Shikra

MP, SP, PSC

Elanus caerulus

Black-Shouldered Kite

MP, SP, PSC

Milvus migrans

Black Kite

MP, SP

Pandion haliaetus

Osprey

SP, SD

Aquila pomarina

Lesser Spotted Eagle

Aquila nipalensis

Pteroclididae

18 19

Accipitridae

DP, MP, PSC

Steppe Eagle

DP, MP, PSC

Falconidae

Falco tinnunculus

Common Kestrel

DP, MP, PSC

Lanidae

Lanius excubitor

Grey Shrike

DP, MP, PSC, PS

Journal of Research in Biology (2014) 4(1): 1228-1239

1236

Koladiya et al., 2014 31

Lanius cristatus

Brown Shrike

DP, MP, PS, SD

Lanius vittatus

Bay-backed Shrike

DP, MP, PS, SD

Lanius schach

Rufous-tailed Shrike

DP, MP, PS, SD

Lanius meridionalis

Southern Grey Shrike

DP, MP, PS, SD

Corvus splendens

House Crow

DP, MP, SP, SD

Corvus macrorhynchos

Jungle Crow

DP, MP, SP, SD

Dicrurus macrocerus

Black Drongo

DP, MP, PS, SD

Saxicola jerdoni

Jerdon's Bushchat

MP, SP, PS, SD

Saxicola caprata

Pied Bush Chat

MP, SP, PS, SD

Oenanthe deserti

Desert Wheatear

Oenanthe picata

Variable Wheatear

SP, PSC, SD

Oenanthe isabellina

Isabelline Wheatear

SP, PSC, SD

Copsychus saularis

Oriental Magpie Robin

DP, MP, SP, PC, SD

Saxicoloides fulicata

Indian Robin

DP, MP, SP, PC, SD

Sternus roseus

Rosy Starling

DP, MP, PS

Acridotheres tristis

Common Myna

DP, MP, PS

Acridotheres ginginias

Bank Myna

DP, MP, PS

MP, SP, SD

Corvidae

Muscicapidae

Sturnidae

Paridae

Parus nuchalis

Pied Tit

Hirundinidae

Hirundo rustica

Barn Swallow

Hirundo smithii

51 52

MP, SP, PSC, SD

SP, SD

Wire-tailed Swallow

SP, SD

Hirundo daurica

Red-Rumped Swallow

SP, SD

Delichon urbica

Northern House-Martin

SP, SD

Pycnonotus cafer

Red-Vented Bulbul

DP, MP, PC, PSC,PS,SD

Pycnonotus leucotis

White-eared Bulbul

DP, MP, PC, PSC,PS,SD

Prinia buchanani

Rufous-fronted Prinia

DP, SP, PSC, PS

Prinia inornata

Plain Prinia

DP, SP, PSC, PS

Prinia sylvatica

Jungle Prinia

DP, SP, PSC, PS

Prinia socialis

Ashy Prinia

DP, SP, PSC, PS

Orthotomus sutorius

Common Tailorbird

DP, MP, PC, PSC, PS

Hippolais caligata

Booted Warbler

DP, SP, PC, PSC, PS

Turdoides caudatus

Common Babbler

DP, MP, PC, PSC, PS

Turdoides malcolmi

Large Grey Babbler

DP, MP, PC, PSC, PS

Turdoides striatus

Jungle Babbler

DP, MP, PC, PSC, PS

Galerida cristata

Crested Lark

SP, PC, PSC

Eremopterix grisea

Ashy-crowned, Sparrow-Lark

SP, PC, PSC

Pycnonotidae

54 55

64 65

1237

Cisticolidae

Sylvidae

Alaudidae

Journal of Research in Biology (2014) 4(1): 1228-1239

Koladiya et al., 2014 66

Mirafra erythroptera

Indian Bushlark

DP, MP, PC, PSC

Mirafra cantillans

Singing Bushlark

MP, SP, PC, PSC

Calandrella raytal

Short-toed lark

MP, SP

Galerida deva

Sykes's Crested Lark

MP, SP, PSC

Nectarinidae

Nectarinia asiatica

Purple Sunbird

DP, SP, PC, PSC, PS

Passeridae

Passer domesticus

House Sparrow

SP, PSC, PS

Anthus rufulus

Paddyfield Pipit

DP, PSC, PS

Lonchura malabarica

Indian Silverbill

Motacilla alba

White Wagtail

SP, PSC

Motacilla flava

Yellow Wagtail

SP, PSC

Motacilla cinerea

Grey Wagtail

SP, PSC

Ploceus philippinus

Baya Weaver

SP, PC

DP, PC, PSC, PS

Alcedinidae

Alcedo atthis

Common Kingfisher

MP, PSC

Dacelonidae

Halcyon smyrnensis

White-breasted Kingfisher

SP, PSC

Cerylidae

Ceryle rudis

Pied Kingfisher

SP,PC, SD

Gruidae

Grus grus

Common Crane

SP, PSC, SD

Grus virgo

Demoiselle Crane

SP, PSC, SD

82 83

Charadridae

Vanellus indicus

Red-Wattled Lapwing

MP, PSC, SD

Anhingidae

Anhinga melanogaster

Darter

PSC, SD

Ardeidae

Bubulcus ibis

Cattle Egret

MP, PSC, SD

Casmerodius albus

Great Egret

SP, PSC, SD

Egretta garzetta

Little Egret

SP, PSC, SD

Mesophoyx intermedia

Intermediate Egret

SP, PSC, SD

Pseudibis papillosa

Black Ibis

MP, PC, SD

Platalea leucorodia

Eurasian Spoonbill

SP, PC, SD

Mycteria leucocephala

Painted Stork

SP, PC, SD

Threskiornithidae

90 91

Ciconidae

MS: Migratory Status, R: Resident, RM: Resident Migratory, WV: Winter visitor, DP: Dense Prosopis, MP: Moderate Prosopis, SP: Sparse Prosopis, PC: Prosopis-Capparis mixed PSC: Prosopis-Suaeda-Calotropis mixed, PS: Prosopis-Salvadora mixed, SD: Suaeda dominant

Journal of Research in Biology (2014) 4(1): 1228-1239

1238

Koladiya et al., 2014 Annexure II. Photographs showing Banni grassland and a few birds sited

Banni grassland

Galerida deva

Accipiter badius

Grus grus

Aquila nipalensis

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Journal of Research in Biology (2014) 4(1): 1228-1239

Journal of Research in Biology

An International Scientific Research Journal

Original Research

Journal of Research in Biology

Determination of age and growth by scale of a population of common trout (Salmo trutta macrostigma, Dumeril, 1858) at the level of Sidi Rachid River (Ifrane. Morocco) Authors:

Abba H 1, Belghity D1, Benabid M2 and Chillasse L3.

Institution: 1. Biology and Health Laboratory. Environmental and Parasitology Team / UFR Doctoral Parasitology compared: Medical and Veterinary Applications." Sciences Faculty. Ibn Tofail University. Kﾃｩnitra B.P. 133, 14000. Morocco. 2. National Center of Hydrobiology and Pisciculture (NCHP) Azrou Morocco.

ABSTRACT:

The determination of age and growth from the scales of trout river (Salmo trutta macrostigma, Dumeril, 1858) at Sidi Rachid River; was employed out of 438 specimens used the size varies between 6.3 cm and 37.5 cm, the relation linking the growth in length of the fish and the growth of the scale. Varied according to the equation Log Lt = 0.8674 ﾃ有og Rt + 0.5349, with a coefficient of correlation( r) = 0.86592138. The period of the end of growth to this population of trout is between December and January, this period is characterized in the middle of the atlas by important reductions in temperature on one hand, the decrease of the network trophique on the other hand which gets coincided with the period of reproduction of the trout. The resumption of the growth is made in a important way from March. The age of the trout's determined by scales varies between 0 + to 5 +. The retro measures are lower than those observed and the equation of theoretical growth of Van Bertalanffy is of the following type: Lt = 34, 96 (1-exp-0,309 (t-0, 27)).

3. Laboratory of biodiversity and wet zones .Uni My Ismail. Faculty of Science. Meknes. Corresponding author: Abba H

Keywords: River trout, age, growth, scales, Sidi Rachid River. Morocco

Email Id:

Article Citation: Abba H, Belghity D, Benabid M and Chillasse L. Determination of age and growth by scale of a population of common trout (Salmo trutta macrostigma, Dumeril, 1858) at the level of Sidi Rachid River (Ifrane. Morocco)

Journal of Research in Biology (2014) 4(1): 1240-1246 Web Address: http://jresearchbiology.com/ documents/RA0414.pdf.

Journal of Research in Biology An International Scientific Research Journal

Dates: Received: 19 Dec 2013

Accepted: 15 Jan 2014

Published: 16 April 2014

1240-1246 | JRB | 2014 | Vol 4 | No 1

www.jresearchbiology.com

Abba et al., 2014 seasonal rhythm with fast growth at the spring and

INTRODUCTION The fishing of salmonids constitutes one of the

summer and a stops its growth during winter period. This

main concerns of the members of fishing associations in

annual growth rate is marked on the various osseous

the nation. Both the common trout (Salmo trutta

structures among which scales are present. The study of

macrostigma, Dumeril, 1858) and the rainbow trout

these osseous structures will allow determining the

(Oncorhynchus mykiss) are appreciated in the fishing

period of the stop of growth and consequently the age

sport. This activity plays an important role in the

and its relation with the size of the specimens of the

socioeconomic development of the region. To alleviate

population of trout in the Sidi Rachid River.

the disappearance of the endemic common trout, the administrators in Morocco resort to the repopulation of

MATERIALS AND METHODS

rivers with vesicle alevins stemming from artificial

Presentation of the environment of study

reproduction which is carried out at the salmon farming station of Ras El Ma.

The environment of study (Figure-1) is Sidi Rachid River, present in the geographical coordinates of

For a long time, numerous studies were

5°9'N N and 33°28'W W. It is at a height of 1620m and

conducted in the determination and knowledge of the

belongs to the rural district of Ait Ali Ouikoub (province

lines of fish the populations of in various aquatic circles.

of Ifrane). The brook is fed by the sources of Sidi Rachid

Besides the parameters size and mass, we also quote the

of which it takes its name with a maximum debit of 172

age of the fish. These various biological lines once

L/S (Abba, 2011) for a main source as well as the waters

determined, can be exploited in the perspectives of

from the station of salmon farming of Ras El Ma (Abba

management

peaches

et al., 2011). From the morphométric point view, the

professionally. The estimation of the age of a fish is of a

River presents a low width which can vary from 2m to 6

big importance to understand the dynamics of a

m, and a depth which does not exceed 1m generally.

population. This determination of the age can be made

Biological material

either in a direct way, or in an indirect way. In this study,

Sampling of fishes

the

various

types

we limited ourselves to one of the direct methods by

The method used in our case is the electric

means of the osseous structures (Spillmann, 1961;

fishing realized by technicians' team of the National

Bagliniere et Maisse, 1990). Although the use of scales

Center of Hydrobiology and Fish farming of Azrou. The

for certain species are questioned (Pikitch et Demory,

number of fish every month varies between 30 and 50

1988), the scales are used for a majority of families with

specimens. For every sinned fish, we have proceed to the

species dulçaquicoles and amphihalines temperate or

measure of its total length (Lt (cm)) with an ichtyometer,

cold regions to be known, almonds, cyprinids and

and before putting it back

precedes (Bagliniere et Lelouarn, 1987; Meunier, 1987;

scales in number from 6 to 20 were removed in the zone

Bouhbouh, 2002). During this study, method used for the

recommended for salmonids according to Ombredane

determination of age and growth of brown trout (Salmo

and Richard(1990). Scales are then tidied up in

trutta macrostigma, Duerile 1858) is by the number, size

envelopes and numbered for further study in the

and pattern of

laboratory with a microfiche (×40).

scales. Indeed, the growth of the

structures mineralized as the scales is proportional to the

in the housing environment,

Determination of the structure of the population

length of the fish (Lea, 1910; Hattour et al., 2005). In

The determination of the number of classes of

temperate zones, the growth of the fish presents a

size of the population of trout at the level of the Sidi

1241

Journal of Research in Biology (2014) 4(1): 1240-1246

Abba et al., 2014

Figure 1: Situation of area of study (Extracted from the map of Azrou. E: 1/50. OOO; division of the map, 1974)

Rachid River during the period of study was made by

extension (AM). The latter is used to determining of the

applying the ruler of Sturge. Number of class = 1 + (3, 3

period of stop of growth. The front of the scale generally

log N), were N is the sample Size.

held to salmonids (Bagliniere et al., 1991) is used for the

Preparation and reading of scales

determination of the total shelf R and other shelves r

The preserved scales dried on the referenced

corresponding to the various annuli, r1, r2, r3 to rn. The

envelopes were taken and rubbed between fingers and

measure was made by means of a graduated ruler on a

cleaned by the water to eliminate any sorts of residues

device microfiche for the same swelling (×42). To work

(Jearld, 1983). The examination of scales can be made by

always in the same condition, the measure of the beam

several ways. The reading chosen in this work as the

was made on the main line, which corresponds to the

determination of the age of the fish was made by a reader

previous field of the scale. The Extension Margin (EM)

of microfiche. The criteria used for the determination of

was calculated according to, Benabid (1990).

rings for the stop of growth vary according to the

Determination of the retro calculation on growth

species. For the salmon kind, the criteria are generally as follows:

The relation binding the size of the fish and the shelf of its scale is linear and is determined by the

Contraction of several circuli in the form of a

following formula (Bryuzgin, 1970): L = b Ra (or Log L

concentric band making the tour (ballot) of the scale

= a Log R + Log b), with, , ‘L’:: length of the fish (cm)

(Bagliniere and Lelouarn , 1987);

in the capture, ‘R’: the previous shelf of the scale of the

Discontinuity of circuli or absence of discontinuity

fish (cm) ie., distance between the center of the scale and

of the circuli in which the thickness decreases;

its outside edge according to a direction strictly constant,

Stepping of the circuli of the annulus on those

‘a’: and , ‘b’: are constants.

previously trained in the side fields either Measures made on scales

The formula of Le Cren (1947) and Philippart, (1975) allows then the retro calculation of the size of the

The rings of ruling of growth allow making

fish every age. Log Ln = Log L + a (Log Rn - Log R).

measurements on the scale to calculate the marginal

With, ‘Ln’ length calculated at the time of the training of the nème ring of the stop of growth in mm; ‘L’: length

Journal of Research in Biology (2014) 4(1): 1240-1246

1242

Abba et al., 2014 observed by some fish in mm; , ‘R’:: length observed by

44 % (Ombredane and Richard, 1990).

the previous beam of the scale in mm; , ‘Rn’: length of

The determination of the period of appearance of

the previous beam of the scale up to the nème ring in

the rings to the stop of growth was made by monthly

mm; and , ‘a’: constant. The theoretical model of growth

analysis of the variations of average Marginal Extension

used is the one of Von Bertalanffy (1938): (Lt = L∞ [1-

(AM) on 387 trout's which presents normal scales.

exp (-K (t-t0))]). (Benabid, 1990;

Bouhbouh, 2002).

During this study, some scales do not present rings on

With K (years- 1): growth rate; L∞ (cm): cut that the fish

the stop of growth; it is about scales of truitelles

in time infinite should have; t0 (years): the age in the

stemming from on-the-spot cross-posted or born alevins

worthless length.

from March, 2007. The (figure-3) shows the results obtained for all the scales of fishes representing stops of

RESULTS AND DISCUSSION

growths.

The histogram of the structure of population of

The analysis of variations of the results showed

the trout (Figure-2) shows a good representation of the

that Marginal Extension presents the minimum only one

individuals and the size of which is between 14 (the

marked well for December and January. This minimum

Middle = 13.8) and 17 cm (the Middle of 16.8).This type

translates not only shows the ring of wintry stop of

of structure is a characteristic of young populations. This

growth but also it corresponds to the period of

structure is explained by the fact that the adults are

heavyweight at the river trout. Indeed this stop of growth

generally fished by farmers in the station of fish farming

is not only due to the period of reproduction which slows

as a source of gametes during the period of artificial

down the growth of the fish but also on the severe

reproduction which comes true in the station of Ras El

conditions which exist during this period of year as the

Ma.

important decrease of temperature and trophiques Among 438 individuals sampled during the

resources (Pourriot and Meybeck, 1995) which are

period of study, the number of river trout presenting

generally due to the snow coverage which knows in this

scales of regeneration is 50 specimens, this constitutes a

region. The resumption of the growth begins gradually

number raised with regard the size of the sample; it is 11,

from February and reaches its maximum during August.

Figure 2: Representation schedules of various classes of common trout and their staff at the level of the Sidi Rachid River during the period of study

1243

Journal of Research in Biology (2014) 4(1): 1240-1246

Abba et al., 2014

Figure 3: Monthly evolution of Average Marginal Extension (AME) of the river trout

This important growth is due to the favorable conditions

The introduction of the coefficient of regression

of the housing environment as the temperature and the

of the relation length (Lt) and length (R) of the scale

abundance of the food reserves, on 438 scales examined

gives the following equation: Log Ln = Log L + 0.8674

(51, scales of regeneration), the age is between 0 + and

× (Log Rn - Log R) (Le Cren, 1947; Benabid, 1990;

4+ for sizes going from 6.3 cm to 37.5 cm. The

Bouhbouh, 2002). The total retro measure lengths from

determination of the size of the trout's at the various

the equation above are listed in the table -1.

moments of their life is based on the principle of

The results obtained for the total retromeasures

proportionality of the growth of the scale with that of its

lengthes are used for the determination of the annual

body. For this end, the equation connecting the previous

average linear increase ( C ) as well as the specific speed

beam R of the scale and the total length (Lt) used in this

of growth noted VSC established by Ricker ( 1958 ):

study was determined as continuation. Log Lt = 0, 8674

C = Ln-Ln-1. (Ln and Ln-1: annual lengthes retro

×Log Rt + 0, 5349. The relation between the total length

measures in time n and n-1 expressed in years. VCS = Ln

of the body of the trout (Lt) and the length of the

-Ln-1 × 100/Ln-1. The obtained results showed that, the

previous shelf of its scale (R) (Figure-4) can be

calculated total retro measures lengths are quite lower

allometrique (Giles and Giguere, 1992).

than the observed annual average lengths. This

Figure 4: Relation between the length of the fish and the previous shelf of its scale at the common trout of the Sidi Rachid River. Journal of Research in Biology (2014) 4(1): 1240-1246

1244

Abba et al., 2014 Table 1: Linear retro measures at the Growth of common trout (combined Sexes) Age Age Observed average group group length (mm) 2008 I 134.60 2007 II 152.40 2006 III 190.09 2005 IV 263.35 2004 V 318.11 Number of fish retro measures Annual average length retro measures Standard deviation Increase in annual average length (mm) Specific speed of growth

I 91.20 103.27 104.71 112.20 128.82 358.00 108.63 13.84 91.20

Length averages retro measures (mm) II III IV 141.25 147.90 165.95 190.54 285.00 161.41 20.04 37.98 44.39

177.07 213.79 234.42 194.00 208.42 29.04 29.17 28.33

245.47 275.42 83.00 260.44 21.17 31.68 14.85

V 309.02 9.00 309.03 33.61 12.20

difference of length can give some explanation by the

of the age. The use of reliable software can give even

fact that the observed average lengths correspond to the

more reliable results for this equation because the sizes

lengths of fish at various moments of the year or the

sinned in other circles sometimes exceed 40cm.

growth is made. On the other hand the total retro measures lengths correspond to the lengths of fish at the

CONCLUSION

time of the training of annuli stag of stop of growth

The use of scales and other osseous structures

during December generally. The average lengths

allow determining particularly the aspects of age and the

observed to both sexes and individuals of the indefinite

analysis of dynamics of a fish population growth. With

sex do not present notable difference for age groups I (1

salmonids, the most recommended method is the scale,

+) and II (2 +). Beyond this age, we notice a variation in

despite some disadvantages such as the difficulty of

favour of females (age groups III (3 +) and IV (4 +)), to

scales reading or the high number of scales of

become slightly raised to the males of age group V (5 +).

regeneration that we obtain. Similarly, the use of another

These variations can be due to the sexual maturity which

method can be very beneficial and will allow having

influences the growth and which is premature in a

more information.

general way at males. Also, the retro measure averages are slightly superior at the females than at the males of the same age group. As for the specific speed of the growth, it is very important for the class II (2 +) and it

ACKNOWLEDGEMENTS I thank the persons in charge of the station of fish farming of Ras El Ma/ Azrou/ Morocco.

exceeded 40 % (combined sexes and various sexes). The decrease is in a very remarkable way as the age of the fish increases and achieves approximately 10 % for fishes of age group V (5 +). For the theoretical model of the growth of Von Bertalaffy (the obtained results watch that the theoretical maximal size of the fish is of L8 = 34.96cm. The theoretical equation becomes then for the population of trout of the Sidi Rachid River is Lt = 34.96 (1-exp-0.309 (t-0.27)). The theoretical length (34.96cm) is lower than the maximal length of the biggest trout (37.5cm) scales of which are used for the determination 1245

REFERENCES Abba H. 2011. Etude écologique et biologique de la truite commune (Salmo trutta macrostigma, Dumeril 1858) dans une rivière de Moyen Atlas (Oued Sidi Rachid) Maroc. Thèse de Doctorat Es Science. Univ. Ibn Tofail. Fac .Sc. Kenitra. Maroc. pp.194. Abba H, Belgghyti D Benabid M, ELIbaoui H, Fadli M and Eby Ould Mohamadou A. 2011. Physicochemical Typology of water of a Middle Atlas River (Morocco) where the common trout (Salmo trutta macrstigma, Duméril, 1858) live: Oued Sidi Rachid. Journal of Research in Biology (2014) 4(1): 1240-1246

Abba et al., 2014 African Journal of Environmental Technology. 5(5): 348-354.

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