Population Ecology of Freshwater Turtle Species of Pothwar Plateau

Home , Brown roofed turtle, Cane turtle, Chelydridae, Geoemydidae, Indian roofed turtle, Pangshura

POPULATION ECOLOGY OF FRESHWATER TURTLE

SPECIES OF POTHWAR PLATEAU

SAIMA YOUSAF 05-arid-342

Department of Wildlife Management Faculty of Forestry, Range Management and Wildlife Pir Mehr Ali Shah Arid Agriculture University Rawalpindi, Pakistan 2017

POPULATION ECOLOGY OF FRESHWATER TURTLE

SPECIES OF POTHWAR PLATEAU

SAIMA YOUSAF

(05-arid-342)

A thesis submitted in partial fulfillment of

the requirements for the degree of

Doctor of Philosophy

Wildlife Management

Department of Wildlife Management Faculty of Forestry, Range Management and Wildlife Pir Mehr Ali Shah Arid Agriculture University Rawalpindi Pakistan 2017

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I dedicate this thesis to my parents. I hope this achievement completes all those dreams they had for me many years ago when they chose to give me the best education they could.

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CONTENTS Page

List of Tables xii

List of Figures xv

List of Abbreviations xviii

ACKNOWLEDEMENT xix

ABSTRACT xx

1 GENERAL INTRODUCTION 1

1.1 INTRODUCTION 2

1.1.1 Global distribution 3

1.1.2 Asian turtle species 4

1.1.3 Turtles of Pakistan 4

1.1.3.1 Freshwater turtles 4

1.1.3.2 Taxonomic classification of freshwater turtles of Pakistan 4

1.1.3.3 IUCN and CITES status of freshwater turtles of Pakistan 5

1.1.4 Physical Characteristics of Freshwater Turtles 7

1.1.4.1 Brown river turtle (Pangshura smithii) 7

1.1.4.2 Ganges soft-shell turtle (Nilssonia gangetica) 9

1.1.4.3 Indus mud turtle (Lissemys punctata) 9

1.1.4.4 Indian roofed turtle (Pangshura tecta) 9

1.1.4.5 Crowned river turtle (Hardella thurjii) 9

1.1.4.6 Spotted pond turtle (Geoclemys hamiltonii) 10

1.1.4.7 Peacock soft shell turtle (Nilssonia hurum) 11

1.1.4.8 Narrow headed turtle (Chitra indica) 11

1.1.5 Basking Behavior 11

1.1.6 Hibernation 12

1.1.7 Food Habits 12

1.1.8 Ecological Importance 13

1.1.9 Threats to Freshwater Turtles 14

1.1.10 Conservation Need 16

1.2 STUDY AREA 17

1.2.1 District Rawalpindi 18

1.2.2 District Chakwal 19

1.3 RATIONALE OF THE STUDY 20

1.4 STUDY OBJECTIVES 20

2 IDENTIFICATION AND DISTRIBUTION OF FRESHWATER TURTLE 22 SPECIES IN POTHWAR PLATEAU

2.1 INTRODUCTON 22

2.2 REVIEW OF LITERATURE 26

2.3 MATERIALS AND METHODS 30

2.3.1 Study Design 31

2.3.2 Materials and Methods 31

2.4 RESULTS 31

2.4.1 Species Diversity 31

2.4.2 Distribution 38

2.4.2.1 Distribution of the Indian flap-shelled turtle (Lissemys punctata) 38

2.4.2.2 The Indian soft shell turtle (Nilssonia gangetica) 38

2.4.2.3 The Brown River turtle (Pangshura smithii) 38

2.4.2.4 Indian roofed turtle (Pangshura tecta) 38

2.5 DISCUSSION 42

2.6 CHAPTER SUMMARY 45

3 POPULATION ESTIMATION OF FRESHWATER TURTLE SPECIES 47 IN POTHWAR PLATEAU

3.1 INTRODUCTION 47

3.2 REVIEW OF LITERATURE 50

3.3 MATERIALS AND METHODS 55

3.3.1 Study Design 55

3.3.2 Capture Mark and Recapture Method (CMR) 55

3.3.3 Direct Count Method 56

3.3.4 Statistical Analysis 59

3.4 RESULTS 59

3.4.1 Population Estimates by Capture Mark and Recapture Method 59

3.4.1.1 Capture, mark and recapture method- turtles abundance 59

3.4.1.1.1 Frequency (F) and density (D) 62

3.4.1.1.2 Seasonal variation CMR method 62

3.4.1.2 Direct Count Method 68

3.4.1.2.1 Abundance of freshwater turtles 68

3.4.1.2.2 Frequency (F) and density (D) of turtles 68

3.4.12.3 Seasonal variation in turtle populations by direct count method 72

3.4.2 Population Estimates of Freshwater Turtles in District Chakwal 72

3.4.2.1 Capture, mark and recapture method-turtles abundance 73

3.4.2.2 Frequency (F) and density (D) 73

3.4.2.3 Seasonal variation in turtle populations (by CMR method) 73

3.4.2.2 Population estimates of turtles using direct count Method 77

3.4.2.2.1 Abundance of freshwater turtles 77

3.4.2.2.2 Frequency (F) and density (D) of turtles 77

3.4.2.2.3 Seasonal variation in turtle populations by direct count method 78

3.4.2.2.4 Comparison of turtle populations by CMR and direct count methods 78

3.4.3.1 Indian flap shell turtles (Lissemys punctata) 78

3.4.3.2 Indian soft shell turtle (Nilssonia gangetica) 88

3.4.3.3 Brown River turtles (Pangshura smithii) 88

3.4.3.4 Indian roofed turtles (Pangshura tecta) 88

3.4.4 Statistical Analysis of Body Parameters of the Turtles 88

3.4.5 Sex Ratios 90

3.5 DISCUSSION 90

3.6 CHAPTER SUMMARY 95

4 INVESTIGATION OF THE FOOD HABITS OF FRESHWATER 97 TURTLE SPECIES OCCURRING IN POTHWAR PLATEAU

4.1 INTRODUCTION 97

4. 2 REVIEW OF LITERATURE 98

4.3 MATERIALS AND METHODS 102

4.3.1 Stomach Contents Flushing 103

4.3.2 Collection of Reference plant Samples 103

4.3.3 Micro-Histological Analysis of Stomach Content Flushing Plants 104

4.3.4 Seasonal Variation in Diet Composition of Each Turtle Species 106

4.3.4.1 Prey species richness index (S) 106

4.3.4.2 Diversity index (H') 106

4.3.4.3 Evenness index (E) 107

4.3.5 Statistical Analysis 107

4.4 RESULTS 107

4.4.1 Collection of Stomach Samples 108

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4.4.2 Stomach Content Analysis 108

4.4.3 Percent Frequency (% F) and Percent Volume (% V) occurrence of 108 Prey Items 4.4.4 Seasonal Variation in Food Consumption of Freshwater Turtle Species 114

4.4.4.1 Indian flap shell turtle (Lissemys punctata) 114

4.4.4.2 Indian soft shell turtle (Nilssonia gangetica) 119

4.4.4.3 Brown river turtle (Pangshura smithii) 119

4.4.4.4 Indian roofed turtle (Pangshura tecta) 119

4.4.5 Prey Species Richness (S), Diversity Index (H') and Evenness Index 120 (E) 4.4.6 Insects Orders Consumed by Freshwater Turtles 121

4.4.7 Plant Species Recorded from Turtles Species Stomach 122

4.5 DISCUSSION 126

4.6 CHAPTER SUMMAY 129

5 GENERAL DISCUSSION 132

SUMMARY 139

LITERATURE CITED 142

APPENDICES 171

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LIST OF TABLES

Table No. Page 1.1 Taxonomic Classifications of Freshwater Turtles of Pakistan 6

1.2 IUCN and CITES status of freshwater turtles of Pakistan 8

2.1 Some details about selected sampling sites in districts (Rawalpindi and 34 Chakwal) of Pothwar Plateau.

2.2 Distribution of freshwater turtle species in selected sampling sites of the two 40 districts (Rawalpindi and Chakwal) of the Pothwar Plateau.

3.1 Relative abundance of four species of the freshwater turtles live captured from 60 twelve (N=12) different sampling sites of district Rawalpindi using Capture, Mark, and Recapture (CMR) method from 2012 to2014(total time spent at each sampling site was 3 hours, numbers of attempts made by using cast net =10 on 1Km transect during all 24 visits).

3.2 Relative abundance of four species of the freshwater turtles live captured from 63 twelve (N=12) different sampling sites of district Rawalpindi using Capture, Mark, and Recapture (CMR) method during summer/spring (total time spent at each sampling site was 3 hours, numbers of attempts made by using cast net =10 on 1Km transect during all 24 visits). 3.3 Relative abundance of four species of the freshwater turtles live captured from 65 twelve (N=12) different sampling sites of district Rawalpindi using Capture, Mark, and Recapture (CMR) method during winter/fall (total time spent at each sampling site was 3 hours, numbers of attempts made by using cast net =10 on 1Km transect during all 24 visits). 3.4 Relative abundance of four species of the freshwater turtles from twelve 67 (N=12) different sampling sites of the district Rawalpindi using “direct count” method from 2012 to 2014. 3.5 Relative abundance of four species of the freshwater turtles from twelve 69 (N=12) different sampling sites of district Rawalpindi using direct count method during summer/spring season.

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3.6 Relative abundance of four species of the freshwater turtles from twelve 70 (N=12) different sampling sites of district Rawalpindi using direct count method during winter/fall season.

3.7 Relative abundance of freshwater turtle species from four selected sampling 74 sites of district Chakwal during 2012-2014 by using CMR method.

3.8 Relative abundance of freshwater turtle species from four selected sampling 75 sites of district Chakwal during summer/spring (total time spent at each site is 3 hours, no of attempts made by net is 10 on 1Km transect visits 24) by using CMR method.

3.9 Population estimates of freshwater turtle species from four selected sampling 76 sites of district Chakwal during winter/fall season (total time spent at each site is 3 hours, no of attempts made by net is 10 on 1Km transect visits 24)by using CMR method.

3.10 Population estimates of freshwater turtles from four selected sampling sites of 79 district Chakwal during 2012-2014 by using direct count method. 3.11 Relative abundance of freshwater turtles from four selected sampling sites of 81 district Chakwal during summer/spring by using direct count method. 3.12 Relative abundance of freshwater turtle species from four selected sampling 82 sites of district Chakwal during winter/fall (total time spent at each site is 3 hours, no of attempts made by net is 10 on 1Km transect visits 24)by using direct count method. 3.13 Comparison of populations of freshwater turtle species by CMR and direct 83 count methods during 2012 to 2014 at sixteen (16) selected sampling sites of the Pothwar Plateau.

3.14 Statistical comparison of turtles populations by using two methods (CMR and 84 direct count) at sixteen selected sampling sites of the Pothwar Plateau by Two- way Analysis of Variance (ANOVA)

3.15 Body measurements of four turtle species captured from all selected sampling 87 sites of Pothwar Plateau (N=2216).

3.16 Statistical comparison of body measurements of four different freshwater turtle 89

specie’s gender collected from Pothowar Plateau by using Two- way Analysis of Variance (ANOVA).

3.17 Statistical comparison of body measurements of four different freshwater turtle 89 species collected from Pothowar Plateau by using Two- way Analysis of Variance (ANOVA).

4.1 Percent frequency (% F) of the prey items recovered from the stomach contents 110 of freshwater turtles collected from selected study sites 4.2 percent volume (% V) occurrence of the prey items recovered from the stomach 110 contents of freshwater turtles collected from the study area 4.3 Seasonal variation in consumption of different food items by four turtle species 115 in the Pothwar Plateau 4.4 Prey species richness (S), diversity Index (H') and evenness index (E) 117 calculated during three different seasons of the year for each of the four turtle species in the Pothwar Plateau 4.5 Percent volume (% V) of insects orders recovered from the stomach contents of 123 freshwater turtle species (N=410) collected from selected study sites.

4.6 Percent volume (% V) of plant species recovered from the stomach contents of 125 freshwater turtle species (N=410) collected from the study area.

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LIST OF FIGURES

Fig. No. Page

1.1 Map of Pakistan showing location of Pothwar Plateau, B) GIS-based map 18 showing location of selected sampling sites in the two districts (Rawalpindi and Chakwal) of the Pothwar Plateau.

2.1 Topographic map showing location of selected sampling sites in the two 36 districts (Rawalpindi and Chakwal) of the Pothwar Plateau (Downloaded and modified from Google Earth). : 1) Loi Bher, 2) Samli TB Sanitorium, 3) Junction of Korang and Sowan Rivers, 4) Bhander, 5) Saroba, 6) New Katarian, 7) Khayaban e Sir Syed, 8) Moti Mahal, 9) Saintha village, 10) Mohra Rajgan, 11) Choha Khalsa, 12) Thoha Bahadur, 13) Jubairpur, 14) Maswal dam, 15) Kallar Kahar Lake, 16) Village Bunn.

2.2 Photographs of freshwater turtle species recorded in the study area 37

2.3 Distribution map of the Indian flap shell turtle (Lissemys punctata) in 41 Rawalpindi and Chakwal districts of the Pothwar Plateau; red dots showing location points where the species was recorded.

2.4 Distribution map of the Indian soft shell turtle (Nilssonia gangetica) in the 41 Pothwar Plateau; red dots showing location points where this species was recorded.

2.5 Distribution map of the Brown River turtle (Pangshura smithii) in water 43 bodies of the two districts (Rawalpindi and Chakwal) of the Pothwar Plateau; red dots showing location points where this species was recorded

2.6 Distribution map of the Indian roofed turtle (Pangshura tecta) in Pothwar 43 Plateau; red dots showing location points where this species was recorded

3.1 A cast net is being cast at Saroba village near Chakari which is associated 57 with Soan river.

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3.2 Photograph of Pangshura smithii showing marginal scute notching process; 57 A) Notching process for marking purpose, B) a notched specimen.

3.3 Taking visual observations of freshwater turtle species at Village Bhandar 58 site near Sihala which is associated (Korang river) transect.

3.4 Graphical representation of seasonal variation in populations of 66

freshwater turtles in district Rawalpindi using CMR method.

3.5 Seasonal variation in populations of freshwater turtles in district Rawalpindi 66 using direct count method

3.6 Seasonal variation in populations of freshwater turtles from district 71 Chakwal using CMR method during the study period

3.7 Seasonal variation in populations of freshwater turtles from district 71 Chakwal using direct count method during the study period

3.8 Comparative populations of freshwater turtles (all species) by CMR and 85 Direct count methods from sixteen sampling sites of Pothwar Plateau during 2012 to 2014.

3.9 Photograph showing body measurments of Lissemys punctata; A) 86 Carapace length, B) Carapace width, C) Plastron width and D) Shell height of Pangshura tecta.

4.1 Photographs taken during the stomach contents flushing procedure,(A) 105 showing stomach contents flushing apparatus (B) Anesthesia injection (C) Injecting the Anthesia into the turtles muscles, (D) Passing Naso-gastic tube into the turtle’s stomach,(E) Filling the turtle’s stomach with water,(F) showing stomach contents being forced out

4.2 Prey items recovered from stomach contents of freshwater turtle species (A) 109 insects body parts (B) earthworms, (C) snails, (D) plant material,(E) algae, (F) soil particles, (G) unidentified material

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4.3 Mean percent volume (%V) occurrence of prey items recovered from the stomach 111 contents of four species of freshwater turtles captured from the study area (A) L. punctata (B) N. gangetica (C) P.smithii (D) P.tecta.

4.4 Two-ways Analysis of Variance (ANOVA) Box plot comparison showed 112 that prey items consumption was significantly different among four freshwater turtle species.

4.5 Two-ways Analysis of Variance (ANOVA) Box plot comparison showed 112 different prey items significantly differ to each other.

4.6 Two-ways Analysis of Variance (ANOVA) Box plot comparison showed 113 different freshwater turtle species significantly differ to each other

4.7 Comparison of prey items consumed by different turtle species in different 116 season (A) L. punctata (B) N. gangetica (C) P. smithii and (D) P.tecta.

4.8 Prey species richness, diversity and evenness indices of four turtle species 118 captured from Pothwar Plateau (A) Lissemys punctata (B) Nilssonia gangetica (C) Pangshura smithii (D) Pangshura tecta

4.9 Comparison of insect orders consumed by freshwater turtles of the Pothwar 123 Plateau values represent mean ± SE

4.10 Photographs of reference and sample slides plant species prepared from 124 recovered plant materials from the stomach contents of freshwater turtle species from the study area (A) Artemisia scoparia (B) Parthenium hysterophorus (C) Withania somnifera (D)Cynodon dactylon (E) Carthamus oxycantha.

4.11 Percent consumption of five different plant species by freshwater turtles in 125 the study area

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LIST OF ABBREVIATIONS

ANOVA Analysis of Variance

CITES Convention on International Trade in Endandered Species of fauna and flora

CL Carapace Length

Cm Centimeter

CMR Capture Mark and Recapture Method

CW Carapce Width

GML General Linear Model

IUCN International Union for Conservation of Nature

Kg Kilogram

Km2 Kilometer square

L Length

L.punctat Lissemys punctata

N. gangetica Nilssonia gangetica

ºC Celsius

(% F) Percent Frequency (% V) Percent Volume P. tecta Pangshura tecta

P.smithii Pangshura smithii

PL Plastron Length

PW Plastron Width

TCF Turtle Conservation Fund

TCM Traditional Chinese Medicines

W Width

AKNOWLEDGEMENTS

First and foremost, all thanks to Allah the Benevolent and Clement, for the blessing and opportunity for me to finish my dissertation. I offer my honorable thanks to Holy Prophet Hazrat Muhammad (S.A.A.W) who is always a source of endless knowledge, guidance, blessing, love and mercy for whole mankind.

I would like to express my deepest gratitude to Dr. Tariq Mahmood

Assistant Professor, Department of Wildlife Management, for helping me to select the topic for my thesis, supervising and guiding me at each step. I value the indefatigable and skillful way in which my thesis was shaped by him. He provided both technical insight and a broad overview essential to this thesis.

My sincere appreciation goes to members of my supervisory committee,

Prof. Dr. Maqsood Anwar, Chairman Department of Wildlife Management and

Dean Faculty of FRM & W, and Prof. Dr. Mirza Azhar Beg, Department of

Zoology, for their valuable suggestions and constructive criticism during my research work.

I wish to offer my heartily and grateful thanks to my friends, Dr. Durr-e-

Shahwar department of Wildlife Management and Khalid Hameed department of

Zoology for their cooperation during the preparation of this thesis.

I express my indeptendness to my husband Dr. Zia-ud-Din Sandhu for their immense understanding and meticulous care. I shall remain indebeted to my

Parents, Brothers and Sisters for their encouragement throughout my educational life.

Saima Yousaf

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ABSTRACT

Freshwater Turtles are the most dynamic group of ‘wetlands’ associated fauna. They form major biodiversity component of aquatic ecosystems, and serve as keystone species benefiting other animals and plants. Pakistan harbors eight freshwater turtle-s species which are facing threat of illegal hunting for their meat and shell. The current study investigated some important ecological parameters including distribution, population, and food habits of freshwater turtle species inhabiting two districts (Rawalpindi and Chakwal) of the Pothwar Plateau. Based on initial field surveys, sixteen water bodies were selected in the study area for data collection. Distribution of freshwater turtle species was determined by employing

“Visual observation” method using binocular, to record the turtle species.

Populations of different turtle species were estimated by using two different methods direct count method and Capture Mark and Recapture (CMR, while their diet composition was investigated by using stomach contents flushing method.

Results showed four freshwater turtle species (Lissemys punctata, Nilssonia gangetica, Pangshura smithii, and Pangshura tecta) distributed in various selected water bodies of district Rawalpindi and two species (Lissemys punctata and

Panshura smithii) in district Chakwal. The Lissemys punctata was the most abundant while Panshura smithii was found the least abundant species.

A total of N = 3004 and N =1681 individual turtles were recorded (by direct count method, and CMR,respectively), belonging to four species from the study area. Average population densityof Lissemys punctata was 86.9/Km², Nilssonia gangetica 36.6/Km², Pangshura smithii 45.5/ Km² and Pangshura tecta 11.2/ Km²

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by direct count method. However, the CMR method used estimated average density of Lissemys punctata as 104.6/ Km², Nilssonia gangetica 47.9/ Km²,

Pangshura smithii 76.2/ Km² and Pangshura tecta 21.5/ Km². In summer/spring season during the study period, turtles populations were high, however, during winter/fall season, turtles populations were found low. Significant differences (p <

0.05) were found between body weights (kg); carapace length and width; plastron length and width, and shell height of of freshwater turtle species and also between the genders of the four species.

The turtles were found omnivorous in their feeding behavior; mean percent volume (% V) of prey items recovered from stomach contents included contributions from insects 26.3 %, earthworms 19.9 %, snails 16.1%, plants 10.2

%, and algae 11.4 %, besides soil particles 7.5 % and some unidentified material.

There was significant differences (p < 0.05) in food consumption during winter, summer and rainy seasons. The consumption of prey items during three seasons was significantly (p< 0.05) different in P. tecta while in rest of three species viz. L. punctata, N. gangetica and P. smithiiit, it differed non-significant ly (p > 0.05).

The current study highlights that four species of freshwater turtles occur in district Rawalpindi and two in district Chakwal of the Pothwar Plateau; Lissemys punctata is most abundant while Pangshura tecta is least common. All four turtlr species are omnivorous in their food habits and their most preferred prey item is insects.

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Chapter 1

GENERAL INTRODUCTION

1.1 INTRODUCTION

Freshwater ecosystem, including rivers and wetlands, provides virtually all easily accessible drinking water on planet and support a wide variety of other species, including fish, amphibians, reptiles and other aquatic organisms and plants. But currently, wetlands are one of the most threatened natural resources

(next to tropical rain forests and coral reefs) wetlands are the most rapidly vanishing habitats (Khan and Ghalib, 2006).

Freshwater animals are the species which generally depend upon freshwater habitats for any critical part of their life cycle (Balian et al., 2008). Turtles are reptiles of the order Chelonia, characterized by most distinct feature, bony or cartilaginous shell developed from their ribs and acting as shield. Freshwater turtles usually have most prominent feature, high domed shells and elephant like feet

(Dubois and Bour, 2010). They live in riverine and wetland habitats, many of which have vanished or been significantly modified in the last 100 years. Human disturbances include habitat alteration, water pollution, terrestrial alteration, channelization, and sand mining, all of which have a negative effect on freshwater turtles throughout the world (Moll and Moll, 2004).

Freshwater turtles are important biodiversity components of the aquatic ecosystems and have much significance due to their ecological position and

economic value. They play the same role in aquatic habitat which vultures play as scavengers in terrestrial ecosystem (TCF, 2002).

Pakistan is a man’s carved northwestern political division of Indo-Pakistan subcontinent and not a natural geological entity. Its varied topography and bioclimate are reflected in the diversity of its soil, climates, habitats, flora and fauna

(Khan, 1980; Khan, 1996; Mufti et al., 1997). There are about 179 reptile species recorded from the country; which comprisies of turtles, tortoises, crocodile, gavial, lizards and snakes (Rehman and Iffat, 1997).

According to IUCN (2009), there are 1,677 reptile species that have been included in the IUCN Red List, with 293 added in 2009; in total, 469 species are threatened with extinction and 22 are already Extinct or Extinct in the wild (Khan et al., 2010).

1.1.1 Global Distribution

Turtles first appeared in the fossil record, belonging to 210 million years ago (late Triassic period) as a very early side-branch of reptilian evolution, and no transitional form exists. The earliest known turtles had distinctive and well developed chelonian features of the carapace and plastron and the shoulder girdle was located within the rib cage (Gaffney, 1990). Globally, turtles have graced the earth for approximately 300 million years. The worldwide distribution of turtles includes, Atlantic Ocean, Gulf of Mexico, Mediterranean Sea, Indo-Pacific regions,

West coasts of North and South America, from central Baja California to Peru,

Tropical regions of the Atlantic, Indian, and Pacific Oceans of the world (Emst et al., 2005). Today, an estimated 220 species of freshwater turtles alone occupy the

water of all vegetated continents from tropics to just outside the Arctic Circle

(Gaffney, 1990).

Globally, turtles are among the most endangered of the major groups of vertebrates, surpassing birds, mammals, cartilaginous and bony fishes, and amphibians. According to the current IUCN Red List (2013), 135 turtle species are officially regarded as globally threatened [(Critically Endangered (CR),

Endangered (EN), or Vulnerable (VU)]. Of the 335 total species of turtles and tortoises, 107 (31.9%) are CR or EN, 167 (49.9%) are Threatened (CR, EN, or

VU), and 175 (52.2%) are threatened or extinct (Van Dijk et al., 2014).

There are 13 families of the Order Chelonia (Testudine), which include land tortoises, freshwater turtles and marine turtles. There are about 289 living species of turtles and tortoises, which are found in different habitat of the World (Azam and Saeed, 2011).

1.1.2 Asian Turtle Species

Asia is rich in biodiversity and correspondingly also rich in turtle species.

Its highest turtle diversity occurs in hotspot regions, the Indo-Gangetic plain, main land Southeast Asia, South China coastal region and New Guinea but almost any area outside the extreme deserts and the altitude and latitude regions is home to some turtle species. At least 100 species of tortoises and freshwater turtles are native to Asia, and new species continue to be described (Van Dijk and Palasuwan,

2000). According to IUCN Red list (2000), out of 89 species of Asian turtles, 87 are listed as threatened, including critically Endangered (CR), Endangered (EN) or

Vulnerable (VU). 21 species are considered (LR: nt) and low risk, least concern

(LR: lc) or data deficient (DD). This is remarkable increase from the 1996 IUCN

Red list, which listed only 33 Asian turtle species on threatened (Traffic, 2004).

1.1.3 Turtles of Pakistan

Pakistan, a division of Indo-Pak subcontinent, has varied topography and bioclimate, which is reflected in the diversity of its soil, climates, habitats, flora and fauna (Khan, 1980; Khan and Khan, 1996; Roberts, 1992 and Mufti et al.,

1997). Till now 167 reptile species have been recorded from Pakistan (Ghalib et al., 1976; Rehman and Iffat, 1997). Turtles of Pakistan are represented by 5 families, 13 genera and 15 species (Khan, 2006).

1.1.3.1 Freshwater turtles

In Pakistan, two families of freshwater turtles are found, namely

Geoemydidae and the Trionychidae; the former family is represented by hard-shell turtles; spotted pond turtle (Geoclemys hamiltonii), crowned river turtle (Hardella thurjii), brown roofed turtle (Pangshura smithii), and Indian roofed turtle

(Pangshura tecta). The latter family (Trionychidae) comprises of soft-shell turtles including Indian narrow-headedsoft-shell turtle (Chitra indica), Indian soft-shell turtle (Nilssonia gangetica), Indian peacock soft-shell turtle (Nilssonia hurum) and

Indian flap shell turtle (Lissemys punctata and ersonii) (Safi and Khan, 2014).

1.1.3.2 Taxonomic classification of freshwater turtles of Pakistan

In Pakistan, freshwater turtles are represented by two Families six genera and eight species (Khan, 2015) (Table 1.1). The freshwater turtle fauna of Pakistan

is entirely Oriental at both the generic and species levels. The family Geoemydidae consists of four species of hard shelled turtles: spotted pond turtle (Geoclemys hamiltoni), crowned river turtle (Pangshura tecta), brahminy river turtle (Hardella thurji) and brown river turtle (Pangshura smithii). These are primarily herbivorous and distributed in Pakistan, India, Bangladesh, Nepal, Burma, Thailand, Vietnam,

Malaysia and Sumatra (Moll, 1987).

The other family, Trionychidae, comprises of four species of soft-shelled turtles; Indian narrow- headed soft shell turtle (Chitra indica), Indian soft shell turtle (Nilssonia gangetica), Indian peacock soft shell turtle (Nilssonia hurum) and

Indus mud turtle (Lissemys punctata). These are larger in size as compared to hard shelled turtles and are primarily carnivorous, distributed in the temperate eastern

North Amercia, Tropical south and Southeast Asia, countries of the Mediterranean region, and Africa (Khan, 1999; 2004; 2006).

1.1.3.3 IUCN and CITES status of freshwater turtles of Pakistan

In Pakistan turtle population rapidly decreasing as there is no protection or conservation law by Government department previously in Pakistan. Consequently, some species populations are becoming threatened. Pakistan is signatory to the

Convention on International Trade in Endangered Species of Fauna and flora

(CITES) since 1976. The Government of Pakistan approved the Federal CITES

Law in May 2012, for effective enforcement of CITES legislations in Pakistan.

Wildlife being a provincial subject, Khyber Pakhtunkhwa Wildlife Department revised the conservation status of freshwater turtles in 2007. The Ganges soft-shell turtle, Peacock soft-shell turtle, spotted pond turtle, and the Indian saw- backed

Table 1.1 Taxonomic classifications of freshwater turtles of Pakistan

Kingdom Phylum Class Family Genera Species

Geoemydidae Pangshura Pangshura smithii (hard-shell Pangshura Pangshura tecta turtles)

Hardella Hardella thurjii

Geoclemys Geoclemys hamiltonii Animalia Chordata Reptilia

Trionychidae Nilssonia Nilssonia hurum (soft-shell Nilssonia Nilssonia gangetica turtles)

Lissemys Lissemy spunctata

Chitra Chitra indica

turtle are included in Appendix-I, while Indian flap-shell turtle is included in

Appendix-II of the Convention on International Trade in Endangered Species

(CITES) (Anonymous, 2005).The Indian soft-shell turtle is Non-CITES species in

Species data base of CITES of 2014 (Anonymous, 2005).

According to IUCN Red List of Threatened Species (2015) status; Narrow- headed soft-shell Turtle (Chitra indica) is endangered; Indian soft-shell turtle

(Nilssonia gangetica) and Peacock soft- shell Turtle (Nilssonia hurum) are

Vulnerable, while one brown river turtle (Pangshura smithii) is Near Threatened.

The majority of the recorded species are unprotected under territorial laws

(PunjabWildlife Acts and Rule, 1974; Islamabad Wildlife Protection, Preservation,

Conservation and Management Ordinance, 1979) while species belonging to families Emydidae, Trionychidae, Varanidae, Euromastycidae and Colubridae enjoy protection under law (Khan, 2015) (Table 1.2).

1.1.4 Physical Characteristics of Freshwater Turtles

1.1.4.1 Brown river turtle (Pangshura smithii)

Brown river turtle (Pangshura smithii) is reported along tributaries of Indus and Ganges rivers (Das, 1999). Its body length may be 23-27cm; males are smaller than females, and have a longer tail with thick base. It mostly lives in flowing waters; however, also been reported from lentic waters (Moll, 1987). The species becomes inactive in winter and hibernates in dry season (Minton, 1996). It feeds on crayfish, crabs, and macrophyte and rotten flesh (Das, 1991), and also on some

Table 1.2 IUCN and CITES status of freshwater turtles of Pakistan

Family Species IUCN Status CITES Status

Geoemydidae Brown River Turtle Low risk Appendix II

(Pangshura smithii)

Geoemydidae Indian Roofed Turtle Low risk Appendix I

(Pangshura tecta)

Geoemydidae Spotted Pond Turtle Vulnerable Appendix II

(Geoclemys hamiltonii)

Geoemydidae Brahminy River Turtle Vulnerable Non listed

(Hardella thurgii)

Trionychidae Ganges Soft-Shell Turtle Vulnerable Appendix II

(Nilssonia gangetica)

Trionychidae Peacock Soft Shell Turtle Vulnerable Appendix II

(Nilssonia hurum)

Trionychidae Indus mud Turtle Low Risk Appendix II

(Lissemys punctata)

Trionychidae Indian Narrow Headed soft shell Vulnerable Appendix II Turtle (Chitra indica)

plant matter and prawn (Moll. 1987). The species is included in CITES Appendix-

II while IUCN (2015) Red list status is low “risk”.

1.1.4.2 Ganges soft-shell turtle (Nilssonia gangetica)

Ganges soft-shell turtle (Nilssonia gangetica) is wide spread in rivers, canals and large lakes throughout Pakistan. Its Carapace is dull olive to green in color, while plastron is ivory, slightly grayish callosities. Its body length may be42-

75 cm and may weigh up to 17 kilograms (Kg). Male is larger than female with longer and thick tail and there is a vent in male near the tail tip. It is extremely apart from animal diet, it also eats pond vegetation and arthropods have also been recovered from its stomach (Das, 1995; Vyas and Patel, 1992). The species is included inCITES Appendix-II while IUCN (2015) Red list status is vulnerable”.

1.1.4.3 Indus mud turtle (Lissemys punctata)

Indus mud turtle (Lissemys punctata) is also widely distributed in Pakistan.

Its shell is light olive brown, plastron creamy; head and limbs gray, with bright yellow spots on head and neck. Male is smaller than female. It may be 17-28 cm in length and its average weight may be up to 7 kg. It prefers water with sand or mud.

It is known to dig in moist soil to escape desiccation (Minton, 1996) and hibernates from November to February during the year. It feeds upon adult frogs, tadpoles, fishes, crustaceans, fish larvae, bivalve mollusks and snail. The species is included in CITES Appendix-II while IUCN (2015) Red list status is vulnerable”.

1.1.4.4 Indian roofed turtle (Pangshura tecta)

Indian roofed turtle (Pangshura tecta) is also widely distributed along the streams connected to river and Indus associated wetlands. The carapace is rich dark brown, in some cases narrowly edged with deep yellow or orange, ventricle keel reddish. Plastron is yellow to orange and the neck is distinctly striped. Mostly, it feeds on vegetable material (Smith, 1931). The species is included in CITES

Appendix-I while IUCN (2015) Red list status is Lower Risk, least concern (LR: lc).

1.1.4.5 Crowned river turtle (Hardella thurjii)

Crowned river turtle (Hardella thurjii) is distributed generally in Indo-

Gangatic plains and East of Brahmaputra system in Pakistan; it is confined to Indus river and its tributaries. There are four yellow- orange s tripes on each side of the head. Carapace is dark brown and dark grey with grey black veritable keel. Feeding habit is omnivorous, mostly feeding on vegetation and rarely found interested in animal food, although a small turtle ate part of a frog (Tikader and Sharma, 1985).

Its IUCN (2015) status is “vulnerable”.

1.1.4.6 Spotted pond turtle (Geoclemys hamiltonii)

Spotted pond turtle (Geoclemys hamiltonii) is restricted to Indo-gangtic

Brahmaputra system, in which it is widely distributed. Only one species is reported from Pakistan. Head is black with yellow spots and numerous white spots represent on the grey neck and limbs. Carapace is elongated with three keels. The color of carapace is black more or fewer wedges shaped. The turtle species is carnivorous feeding readily on fishes, insects and snails but not on vegetation (Minton, 1966). It is included in CITES Appendix-II, while IUCN (2015) reports it as “vulnerable”.

1.1.4.7 Peacock soft shell turtle (Nilssonia hurum)

Peacock soft shell turtle (Nilssonia hurum) is distributed throughout India,

Bangladesh and Pakistan, it is known from the Indus and its tributaries. Carapace olive with yellow rim and maximum length of carapace 40-60cm. Head and limbs olive, head with dark reticulation and yellow blotches on snout. Plastron colour is light gray. Sexually dimorphic, male with longer and thicker tail, anal aperture close to the tail tip (Grey, 1931).It is included in CITES Appendix-II, while IUCN

(2015) reports it as “vulnerable”.

1.1.4.8 Narrow- headed soft shell turtle (Chitra indica)

Narrow- headed soft shell turtle (Chitra indica) is widely distributed in the river systems of the Oriental region, right from Thailand to Pakistan, where it is common in the Indus and its tributaries. Its maximum body size 35-115 cm, weight

120 kg. The Carapace is dull olive or bluish gray, with a pattern of wavy reticulations, neck and forelimbs with similar pattern. Clutch size ranges from 60 to

120 eggs (Gray, 1931). It is included in CITES Appendix-II, while IUCN (2015) reports it as “vulnerable”.

1.1.5 Basking Behavior

Many freshwater turtles during winter, when the temperature of the water becomes low, bask in the sunlight. Due to cooler weather and cold water, this cold blooded reptile avoids living in water and so they come outside the lake to enjoy sunshine to keep them warm (Manning and Grigg, 1997). There are several advantages of this basking behavior; 1) basking allows the turtle to absorb heat,

heat allows the body to work at maximum capacity, 2) turtle gets Vitamin D from sun light which is necessary requirement for absorption of Calcium, as Calcium is fundamental element of freshwater turtle shell (Lovich, 1988).

1.1.6 Hibernation

Many of the freshwater turtle species hibernate during winter when temperature of water body drops very low. They put them in a dormant state, where they conserve energy during a time when they will not eat, but reduce their oxygen intake, and keep them safe from freezing. During the waning weeks of summer, the turtle eats voraciously to store adequate energy for the winter months. When the temperatures start to drop at night, the turtle stop eating to allow for the digestive tract to get empty. If food is present during hibernation it will not be digested and may cause illness as well. They may awaken and move a bit at times when the temperatures are warmer, but this is minimal. In captivity, many of these turtles retain the instinct to hibernate. Cooler nights and later cooler days, along with shortened day length clues them that winter is coming (Jackson, 2002).

1.1.7 Food Habits

The freshwater turtles play an important role in the aquatic ecosystems by feeding on different slow moving aquatic animals like crabs, snails, insects, dead animals and their fragments, thus reducing the water pollution (Hossain and Sarker

1993). They perform a valuable service as scavengers in the tanks, rivers and stagnant water and thus keep the aquatic systems free from pollution (Rao, 1986).

Turtles are very important scavengers, herbivores, and carnivores and form valuable biomass of an ecosystem. They form an important link for plants by

providing dispersal mechanism and so contribute to environmental diversity and develop symbiotic association with a diverse array of organisms (Brooks et al.,

1988; Lovich, 1994). Most of the juvenile turtles are herbivorous but become omnivorous as growth proceeds (Georges, 1982). Turtles clean up water resources by scavenging on dead organic matter and this regard help maintain healthy populations of fish (Lovich, 1994).

Several studies indicate that the different populations of the same turtle species have different feeding habits, related to either habitat quality or displacement in the feeding niche. In the wild, turtles eat small crustaceans, fish, worms, aquatic insects and water snails and frogs eggs. As they have no tongue or teeth, they can only eat underwater (Georges et al., 1986).

The food of fresh water turtles depends upon the species because turtles show both feeding behavior; omnivorous and carnivorous. They have varied diet comprising of shrimp, fish, shellfish, vegetables and supplements. They need a lot of calcium to develop their shell. Fish and mussel meat is a good food source for turtles (Moll, 1976; Bury, 1979; Gibbons et al., 1979).

1.1.8 Ecological Importance

Freshwater turtles are important component of the biodiversity of the aquatic ecosystems and have much significance due to their ecological position and economic values. Although not as obviously visible as birds or ecologically dominant as large mammals or fish, turtles often fulfill important role in the ecosystem, including seed dispersal and vegetation management, control of insect and snail populations, and keeping water clean and population healthy by

scavenging dead animals and preying upon week and sick individuals (Turtle

Conservation Fund, 2002).

Turtles are exploited as a food source in many parts of the world.

Freshwater turtles are consumed by a group of people as a source of protein for its delicacy (Rao, 1987). The demand of turtles is increasing progressively as their meat and other productive are very delicious. Rare turtle species are also collected for food (Zhao and Adler, 1993).

Medicinal value of turtles makes them very important creature. Traditional

Chinese Medicine (TCM) is prepared from soft shell turtles. The practitioners of these medicines believe that the turtle shells are highly effective for purifying blood and also for curing many diseases (Zuberi, 2007). They are also considered as most palatable non- marine Chelonians in Southeast Asia (Jenkins, 1995). The shell of some freshwater turtle species is highly prized for its ornamental value and some in areas it utilized in making tools and musical instrument by tribal people.

Hatchlings of certain species fetch high prices in the pet trade and are easily smuggled worldwide (IUCN/SSC, 1996). Bangladesh exported approximately

3164.24 metric tons of freshwater turtles in the fiscal years 1980-2002

(Anonymous, 2002). Due to over exploitation and habitat destruction, turtle populations are declining (Rao, 1986). Unsustainable exploitation is of a great threat of extinction in near future (Sarker and Hossain, 1997).

1.1.9 Threats to Freshwater Turtles

Populations of many species of freshwater turtles are declining worldwide

(Gibbons et al., 2000). Turtles face an increasingly serious set of threats to their

survival making them one of the most severely threatened vertebrate clade

(Klemens, 2000).

One of the major threats to freshwater turtles in Pakistan includes their importance abroad as they have become hot smuggled items. In the provinces of

Sindh and Punjab, fishermen illegally hunt turtles and in some cases, a turtle is killed for a mere payment of 2-4 US dollars. The bulk of turtle export, worth millions of dollars, is destined for Thailand, Singapore, Hong Kong, South Korea,

China and Taiwan, where their soft shells and chest pellicles are used to manufacture Traditional Chinese Medicines. Main centers of turtle pet trades are

Jakarta, Singapore and Bangkok. In Pakistan, at present, there is a ban on export of all the reptiles including turtles under various legislations (Zuberi, 2007).

There are two main types of trade in turtles that occurs in Asia. One is high volume commodity trade in whole live turtles or turtle parts for consumption; the other is the pet trade. Hunting of turtles for sustenance might have less impact, but the growing demand in turtle plastron in China for making gelation has up scaled the hunting to an unsustainable level. The high income from turtle plastron has attracted other communities to this trade those who were not engaged traditionally in hunting turtles (Asian Turtle Trade Working Group, 1999).

Habitat destruction is also a major threat to the tortoises and freshwater turtles. Due to habitat destruction magnified changes to their habitat resulting from human activities. From logging to slash-and-burn agriculture to pollution, plus the damming and channeling of rivers and the landscapes, turtles call homes, are being damaged (Doupe et al., 2009).

Hatchlings of certain species fetch high prices in the pet trade and are easily smuggled worldwide (IUCN/SSC, 1996). Bangladesh exported approximately

3164.24 metric tons of freshwater turtles in the fiscal years 1980-2002

(Anonymous, 2002). Due to over exploitation and habitat destruction, turtle populations are declining (Rao, 1986). Unsustainable exploitation is of great threat of extinction in near future (Sarker and Hossain, 1997).

1.1.10 Conservation Needs

As a result of significant threats, some species of freshwater turtles are becoming threatened. The Ganges soft shell turtle, peacock soft shell turtle, spotted pond turtle, and the Indian saw-backed turtle are included in Appendix-I, whereas,

Indian flap-shell turtle is included in Appendix-II of the Convention on

International Trade in Endangered Species (CITES) (Anonymous, 2005). The

Indian soft-shell Turtle, Brahminy turtle and Smith’s terrapin are non-CITES

(Anonymous, 2005).

In Pakistan, conservation of a wildlife species is provincial responsibility.

Each of the four provinces and the capital territory of Islamabad has its own

Wildlife Conservation Legislations. A federal ban was imposed in August 1981 on the export of all wild mammals, reptiles and certain birds including their parts, products and derivatives, except for limited numbers of authorized hunting trophies.

Turtles are considered to be of particular conservation concern because their life history includes low reproductive output, late maturity, and habitat

requirements of wetlands and terrestrial environments (Congdon et al., 1996,

Klemens, 2000). Many turtle and tortoise species depend on high adult survival to off set high egg and juvenile mortality in the wild (Schlaepfer et al., 2005).

Removing even small fractions of adults from a population can cause declines or delay a population recovery (Congdon et al., 1994, Heppell, 1998). Soft shell turtles are among the most threatened groups of freshwater animals that are in need of urgent conservation attention (Kumar et al., 2009).

1.2 STUDY AREA

The current study was conducted during March 2012 to Feburary 2014, in the Pothwar Plateau located at 33º30"0' N to and 73°0" 0' E . The Plateau comprises of four districts, namely Jhelum, Chakwal, Rawalpindi and Attock

(Fig.1.1). The Pothwar Plateau is bounded in the east by Jhelum River, in the west by Indus River, in the north by Kala Chitta Range and Margallah Hills, and in the south by the Salt Range. The approximate area of the Plateau is approximately 2.2 million hectares. Dominant habitat type in the area is dry sub-tropical, semi- ever green scrub forest (Roberts, 1991). The Pothwar Plateau harbors wide spectrum of wildlife species, including Punjab Urial (Ovis vignei punjabiensis), Chinkara

(Gazella bennetti), Indian pangolin (Manis crassicaudata), two species of mongooses, (small Indian mongoose Herpestes javanicus and the grey mongoose

H. edwardsii), chukar partridge (Alectoris chukar), grey francolin (Francolinus pondicerianus), see-see partridge (Ammopaerdix griseogularis) and black francolin

(Francolinu sfrancolinus) (Awan, 1998). Important carnivores include Indian wolf

(Canislapus pallipes), jungle cat (Felischaus), Asiatic jackal (Canus aureus), found

Figure 1.1 A) Map of Pakistan showing location of Pothwar Plateau, B) GIS-based map showing location of selected sampling sites in the two districts (Rawalpindi and Chakwal) of the Pothwar Plateau.

found here include Indian wolf (Canislapus pallipes), Jungle cat (Felischaus),

Asiatic jackal (Canus aureus) red fox (Vulpus vulpus), and yellow throated martin

(Martes flavigula) (Hasnain, 1985).

The climate of the area is sub-humid, sub-tropical continental type. There are two distinct rainy seasons; the summer season or the monsoon rains start by about mid-July and last until the mid of September. Most of the precipitation is received during July and August. The winter rains begin in January and persist up to early March. The maximum temperature is 45oC during summer but drops below

0oC during winter, January being the coldest and June the hottest month of the year

(Awan, 1998).

1.2.1 District Rawalpindi

District Rawalpindi is located in the northern most part of the Punjab province; it features a humid sub-tropical climate with long and very hot summers, a monsoon and short, mild, wet winter season. The average annual rainfall is

990 mm, most of which is received during the summer monsoon season. In summer, the maximum temperature can sometimes soar up to 52 °C, while it may drop to a minimum of−4 °C in the winter (Ahmad et al., 1990). It covers an area of

5286 Km² (Anonymous, 2007).

1.2.2 District Chakwal

District Chakwal borders the Rawalpindi and Attock districts in the north, district Jhelum in the east, district Khushab in the south and district Mianwali in the west. The total area of district Chakwal is approximately 6,524 square kilometers.

The district is also semi-arid area with a shortage of irrigation systems and water sources for agriculture (Ahmad et al., 1990). The total area of Chakwal district is

6,609 square kilometers (http://en.wikipedia.org/wiki/Chakwal_District).

Ecological data are lacking about different species of freshwater turtles in the country, therefore, the current study was designed to investigate the distribution, habitat association, population, and food habits of freshwater turtle species in two districts (Rawalpindi and Chakwal) of the Pothwar Plateau.

1.3 RATIONALE OF THE STUDY

Freshwater turtles play important role in aquatic ecosystems by being omnivores, carnivores and scavengers and so reduce water pollution. Moreover, this group is utilized in preparing Traditional Chinese Medicines (TCM), which is thought to be helpful in purifying blood and curing many other diseases. They also possess ornamental values. For conservation and management purposes, it is pre- requisite to have baseline information about biology and ecology of a species, including habitat requirement, distribution, population estimates and feeding habits

In Pakistan, formal scientific studies on freshwater turtles are very scanty; a few published have reported checklists and their distribution in some parts of the country. However, studies related to the ecological parameters of freshwater turtles like habitat requirements, food habits etc are lacking. Therefore, the current study was designed to investigate the distribution, population, and food habits of freshwater turtle species inhabiting the Pothwar Plateau.

1.4 STUDY OBJECTIVES

The current study was designed to focus on freshwater turtle species inhabiting

two districts (Rawalpindi and Chakwal) of the Pothwar Plateau with following main objectives:

• To identify different freshwater turtle species occurring in the study area

and determine their distribution.

• To estimate populations of various turtle species inhabiting the study area.

• To investigate food habits of freshwater turtle species occurring in the study

area.

Chapter 2

IDENTIFICATION AND DISTRIBUTION OF FRESHWATER

TURTLE SPECIES IN POTHWAR PLATEAU

2.1 INTRODUCTION

Freshwater turtles are one of the most important groups of wetlands associated fauna. They play the similar role in the aquatic ecosystem as is played by vultures in the terrestrial habitats. Freshwater turtles have been documented to live up to 50 years and commonly they live to 20 years life span. Mostly vegetarian, but there are some species that depend heavily on meat and some others are essentially carnivorous in food habits. These possess high, domed shells that are hinged so the animals can completely enclose it self to escape predation

(Klemens, 2000).

Turtles comprise a familiar group of vertebrates that occupy a broad range of habitats and ecological functions. Some species are important in energy flow, and may dominate vertebrate biomass, particularly in aquatic habitats (Congdon and Gibbons, 1986). By virtue of their trophic position, biomass, and vulnerability to anthropogenic perturbations, reptiles can be important indicators of environmental quality (Gibbons and Stangel, 1999). Due to their longevity and habits, turtles do accumulate environmental contaminants and are ideal for monitoring their presence in aquatic environments (Golet and Haines, 2001).

The species richness of turtles and tortoises in the world that have existed in modern time (since 1500 AD), and currently generally recognized as distinct

consists of approximately 324 species and 140 additional sub species or 464 total taxa and of these, 10 taxa have gone extinct (Rhodin, 1994).

Freshwater turtles are divided into four families, each comprising of multiple species featuring unique identification characteristics. Approximately 95 species make up the Emydidae family of freshwater turtles commonly known as

“box” and “water” turtle, the members of Emydidae being extremely diverse. The yellow belly turtle can reach up to 28 cm long and sports vertical yellow marking on its carapace, otherwise known as hard out shell and yellow marking on either side of its head. A smaller member of its family, the chicken turtle (Deirochelys reticularia) is between 10-15 cm long, with prominent yellow striping down its over legs. All species in the Emydidae family have hard domed or low-arching carapaces as well as hardened bottom shells (Legler, 1980).

The second family of soft shell turtles is Trionychidae. Instead of the hard, domed carapaces of other freshwater turtles, these turtles feature soft, leathery outer shells. Most of the species in this family also possess elongated snouts with fleshy lips covering their beaks, and a flattened appearance. The Florida soft shell

(Apalone ferox) has uniformly-colored olive green to brown, oval-shaped shell

(Legler, 1980).

The family Chelydridae mostly comprises of snapping turtles having very large heads and strong jaws; the upper jaw being hooked. Snapping turtles have a surly disposition, which couples with powerful jaws and clawed feet make for a fearsome predator. The common snapping turtle Chelydra serpentina is dark olive brown, can weigh up to 35 pounds and loves to hide in the mud waiting to ambush

its prey insects, amphibians, reptiles, birds and even small mammals (Le DienDuc and Broad, 1995).

Family Kinosternidaeis known as musk or mud turtles, indigenous to the

Americas. Aquatic environment is most preferable habitat. These freshwater turtles have the ability to release a strong, unpleasant scent when disturbed; they feature glands on either side of the body, where the soft bridge or “neck” connects to the shell. Eastern musk turtle (Sternotherus odoratus) is a small species having 2-to-4- inch highly arched, olive brown or black carapace and small spiky organs on the chin and throat (Wilson and Swan, 2003).

About 210 million years ago (late Triassic period), the fossil record of turtles first appeared as a very early side-branch of reptilian evolution and no transitional form exists. Turtles shell is one of the most existence morphological innovations in vertebrate history. The earliest known turtles had distinctive and well developed chelonian features of the carapace and plastron and the shoulder girdle was located within the rib cage. It is the single unifying theme for the great diversity of turtle species that come to occupy our oceans, forests, swamps, lakes, rivers and deserts. An estimated 220 species of freshwater turtles alone occupy the waters of all vegetated continents from tropics to just outside the Arctic Circle. The earliest known turtles had distinctive and well developed chelonian features of the carapace and plastron and the shoulder girdle was located within the rib cage

(Gaffney, 1990).

The distribution of freshwater turtle species is known to their significant relation with habitat attributes (Froese and Burghardt, 1975; Major, 1975; Ernst et

al., 1994). Habitat is most fundamentally defined as the place an animal inhabits.

More rigorous definition that ties habitat with a particular species is that it must possess a set of resources and environmental conditions that allow occupancy, and reproduction of that particular species (Morrison et al., 1992).

Freshwater turtles inhabit different water bodies ranging from shallow ponds to deep lakes and rivers, they are divided into two broad categories namely hard-shell turtles; Emydidea turtles, and soft-shell turtles; Trionychid turtles

(Smith, 1933; Pritchard, 1979; Daniel, 1983; Moll, 1984; Das, 1985). Some genera are distributed very widely but others are restricted to small continental areas. Two families of freshwater turtles (Geoemydidae and Trionychidae) are reported to occur in the rivers of the Punjab, Pakistan (Anonymous, 2005).

Ecologically, turtles are found in different habitats; some occurring in still water (like ponds, lakes) while others are found in running water such as rivers, canals, streams and so on. A few species come on land primarily for egg laying

(Holland and Bury, 1998). Soft shell turtles are highly aquatic and are also capable of rapid locomotion on land as well as in water. Their skin is soft with scales which form a few sickle shaped lamellae on limbs after reduction (Ernst et al., 1994).

Earlier studies on chelonians show that most fundamental habitat of family

Trionychidae; soft shell turtle species Nilssonia gangetica and Nilssonia hurum is running rivers, lakes and ponds, Chitra indica mostly found in lotic sandy stretches of rivers and is known to be a carnivore (generalist), Pangshura tentoria has a wide range of habitats, that is, it prefers large running water to small streams and is

herbivore (generalist) feeder (Moll, 2004).

There are some fundamental anthropogenic threats which have significant effect on distribution of freshwater turtle species; Urbanization has led to the homogenization, fragmentation, isolation and degradation of physical environments with significant impact on local biodiversity Urbanization has led to the homogenization, fragmentation, isolation and degradation of physical environments with significant impact on local biodiversity (Forman et al., 2003; McKinney,

2006). Habitat fragments; small habitat patches, landscape and isolation often dispersed widely populations within relatively of that habitats (Hanski, 1999;

McKinney, 2006). Isolation in urban areas results in decreased movement between sub-populations and increases the risk of population extinction (Hanski, 1999).

Many habitat fragments in urban areas are not large enough to support populations of some species (Chelazzi et al., 2007).

Turtle populations appear to be declining globally, due to the combined impacts of habitat loss, degradation, and fragmentation, road kill, invasive species, pollution, disease, unsustainable harvest, and global climate change (Gibbons et al.,

2000, Klemens, 2000).

2.2 REVIEW OF LITERATURE

Approximately 300 extant species of freshwater turtles and tortoises are distributed over 7 major geographic regions worldwide, Asia being the region where there is most species diversity found, however, the region also have the greatest percentage of threatened species, with more than 75% Critically

Endangered, Endangered, or Vulnerable, and 91% included in the IUCN Red List

(Turtle Conservation Fund, 2002).

Freshwater turtles inhabit a great variety of habitats including terrestrial, semi-aquatic and aquatic systems (Bedoya-Gaitan and Godoy, 2008). Freshwater turtles in India inhabit different water bodies ranging from shallow ponds to deep lakes and rivers. The Ganga River is a major river in India, which is a home for different species of aquatic animals including freshwater turtles. In addition, major tributaries of the Ganga River including Chambal, Yamuna, Ken, Son, Ghagra,

Girwa and Ramganga also hold different species of turtles. Few studies conducted on Indian freshwater turtles have mainly dealt with taxonomy and their broad distribution ranges (Smith 1933; Pritchard 1979; Daniel 1983; Das 1985; Tikader and Sharma, 1985).

Freshwater turtles form an important component of aquatic ecosystem, in

Pakistan, different species being distributed in different areas (Khan, 2006).

Turtle’s distribution is not uniform, with aggregations occurring in ponds. Mostly turtle’s distribution related with basking sites at a pool due to this geographical competition increases as a result of limited resources available (Bury, 1972). The turtle fauna of the Mae Klong Basin (Thailand) may be one of the most diverse assemblages in the world (Gibbons, 1990).

In Pakistan, two families of freshwater turtles, namely, Geoemydidae and

Trionychidae occur in the rivers, pond and other water bodies. These two families include eight different freshwater turtles species; Chitra indica (Narrow-headed

Soft-shell Turtle), Geoclemys hamiltonii (Spotted-Pond Turtle) Nilssonia gangetica

(Indian Soft-shell Turtle), Nilssonia hurum (Peacock Soft-shell Turtle), Lissemys

punctata (Indian Flap-shell Turtle), Pangshura smithii (Brown River Turtle),

Pangshura tecta (Indian Saw backed Turtle) and Hardella thurgii (Brahminy River

Turtle) (Anonymous, 2005).

In Punjab province of Pakistan, five rivers, seven irrigation canals were surveyed form 19 different points for identification and distribution of freshwater turtle species. Basking turtles were directly counted, and identified with the help of

Binocular Minolta (10x50 mm). Eight freshwater turtle species found include

Brown River Turtle (Pangshura smithii); Indian Saw-backed turtle (Pangshura tecta); Brahminy River Turtle (Hardella thurgi); Spotted Pond Turtle (Geoclemys hamiltonii), Indian Soft-shell Turtle (Nilssonia gangetica); Peacock Soft-shell

Turtle (Nilssonia hurum), Narrow-headed Soft-shell Turtle (Chitra indica), Indian

Flap-shell Turtle (Lissemys punctata) (Akbar et al., 2006).

Visual obersation and cast net method was applied in Korang river,

Rawalpindi/Islamabad for distribution and relative abundance of freshwater turtle species. Three species of freshwater turtles; Indus mud turtle (Lissemys punctata), the Indian soft-shell turtle (Nilssonia gangetica) and the Brown river turtle

(Pangshura smithii) distributed in Korang River, Rawalpindi/Islamabad (Mahmood et al., 2012).

Six species of freshwater turtles including Pangshura smithii, Pangshura tecta, Hardella thurgi, Chitra indica, Nilssonia gangatica and Lissemys punctata were found in the River Indus, adjacent to Sukktar Barrage, Guddu Barrage,

Jamaldin Wali and pond areas adjacent to Kandhkot. Study was based on direct observation depend on basking and floating turtles. The Chitra indica and

Pangshura smithii were found to be abundant in various parts of the River Indus.

(Azam et al., 2005).

Some already published records about reptiles and freshwater turtles from

Pakistan include Minton (1966) who published a book titled “Herpetology of

Pakistan” in which he described the distribution of the reptiles of Pakistan including turtles. Khan and Mirza (1976) published a key and checklist of Reptiles

Chelonian including distribution of turtles in Pakistan. Ghalib et al., (1981) published checklist of reptiles of Pakistan including their known distribution. Azam et al., 2005 published a paper entitled “Some observations on the distribution and abundance of freshwater turtles in river Indus”.

If we look at the worldwide of freshwater turtle species, Lissemys punctata is distributed in Bangladesh, India, Myanmar, Nepal and Pakistan, Main habitat of this species where it was distributed in the Indian sub-region is the Indus basin of

Pakistan through India (Bhupathy et al., 2011). The species also occurs at 1000 m above sea level. Occurrence of L. punctata is reported from every region of Nepal specifically (Das 1991, 1995, 2001). The species is recorded from the Brahmaputra flood plains Assam, India (Talukdar, 1979). In Pakistan, Lissemys punctata occurs from Indus Drainage to delta, while in India, and Bangladesh its records come from

Ganges River (Minton, 1966).

Indian flap shell turtle (Lissemys punctata) is reported from Bangladesh, from different types of habitats. There are nine categories of habitats that were studied, Marshland, agriculture fields, streams, canals, tanks, derelict ponds and domestic ponds. Most of the time, the turtle species was found in burrowing condition, while it was observed rarely in feeding mode.

Distribution of Pangshura smithii includes its records from Northern India,

Pakistan, Nepal and Bangladesh. In Pakistan, this turtle species is mainly confined to middle Sind, also been reported from Brahmaputra and Ganges Rivers in India,

Bangladesh and also in the Mahanadi Basin (Ghalib et al., 1976).

Das et al., (2011) conducted a study by sending 129 questionnaires to the local fisheries offices, Department of Fisheries, Agricultural and Cooperative

Ministry, in addition, data of live specimens and preserved specimens collected.

The Indian soft-shelled turtle (Nilssonia gangetica) and Indian peacock turtle

(Nilssonia hurum) is distributed, over eastern Pakistan northern and central India,

Bangladesh and Nepal (Das et al., 2011). Tributaries of the rivers Indus, Ganga,

Brahmaputra and Subarnarekha as well as their numerous tributaries some verified examples have come from these areas, as well as from isolated bodies of water, sometimes far from major river drainages (Smith, 1931; Moll et al., 1986).

The Ganges soft-shelled turtle (Nilssonia gangetica and Nilssonia hurum) distributed in the Ganges, Indus, and Mahanadi river systems of Pakistan, northernIndia, Bangladesh, and southern Nepal. Preferable habitats of Ganges soft shell turtles include deep rivers, streams, large canals, lakes and ponds, with a bed of mud or sand. Mostly it inhabits that area where the water is turbid. The species spends more time eating aquatic plants and a large variety of smaller animals, such as fish, mollusks, insects, amphibians, and waterfowls (Bonin et al., 2006).

In Pakistan, protection of wildlife is a provincial responsibility, North-West

Frontier Province (NWFP) Wildlife Department undertook the initiative to protect freshwater turtles in the province after realizing the sensitivity of the issue, On

August 22nd, 2007, the NWFP Wildlife Protection Act, 1975, was amended and all the freshwater turtles of the province were included in the list of protected animals.

On November 30th, 2007 Punjab Wildlife Department revised the Punjab Wildlife

Protection Act 1974 and made an amendment to include the Order Chelonia

(Turtles and Tortoises) in the list of protected animals (Schedule III) (Baig et al.,

2008). However, some species populations are becoming threatened; the Ganges soft-shell turtle, Peacock soft-shell turtle, spotted pond turtle, and the Indian saw- backed turtle are included in Appendix-I, whereas, Indian flap-shell turtle is included in Appendix-II of the Convention on International Trade in Endangered

Species (CITES) (Anonymous2005). The Indian Soft-shell Turtle, Brahminy

Turtleand Smith’s terrapin are non-CITES (Anonymous, 2005).

2.3 MATERIALS AND METHODS

2.3.1 Study Design

A reconnaissance survey of the potential water bodies of the Chakwal and

Rawalpindi districts was conducted to record the presence of freshwater turtles.

Based on the survey 16 sampling sites (water bodies) were selected 4 from district

Chakwal and 12 from district Rawalpindi (Table 2.1). The selected water bodies, included, both running (river, steam, nullahs) and standing (ponds, lakes) water bodies. The sampling sites of the Rawalpindi district were further divided into sub- sampling sites where the water body was a long river, for data collection. Each sampling site was a one kilometer long transect with a width of 0.25 m either site.

2.3.2 Materials and Methods

We surveyed the selected sampling sites (water bodies; such as water

ponds, dams, nullahs, lakes and rivers; N=16) in the two districts (Rawalpindi and

Chakwal) of the Pothwar Plateau during 2012 to 2014. To identify and enlist different freshwater turtle species in the study area, we applied “cast net” to capture and identify different turtle species from selected sites. The captured animals were released back safely. “Visual observation” method using binocular, by walking along the one kilometer long transect (in case of flowing water body like river or stream) and recording number of turtle species sitting on the bank, on a stone in water or on any floating object (in case of standing water body like pond, and lake).

The turtles with their heads out of water were also counted. During visual observations, still camera photographs of the turtle species were taken when flushing to identify the turtle species. The species was identifie by matching photographs of identification species available in literature and by using (Khan

2006; Khan, 2015) keys.

Information was also collected from local people of the area about occurrence of different turtle species in the area by showing them photographs of various turtle species. Geographical coordinates of the sampling sites having turtle populations were recorded using GPS used to construct a distribution map of different freshwater turtle species in the study area occurrence of different turtle species.

2.4 RESULTS

2.4.1 Species Diversity

Four species of freshwater turtles were recorded from the study area Indus mud turtle (Lissemys punctata), Indian soft shell turtle (Nilssonia gangetica),

brown river turtle (Pangshura smithii) and Indian roofed turtle (Pangshura tecta)

(Fig. 2.1). Two species were found in district Chakwal; Indus mud turtle (Lissemys punctata) and brown river turtle (Panshura smithii) while four species were found in district Rawalpindi including Indus mud Chakwal; Indus mud turtle (Lissemys punctata) and Brown river turtle (Panshura smithii) while four species were found in district Rawalpindi including Indus mud turtle (Lissemys punctata), Indian soft shell turtle (Nilssonia gangetica), Brown river turtle (Panshura smithii) and Indian roofed turtle (Pangshura tecta) due to its Plastron with epidermal moveable flanges, seven plastral Callosities (Khan, 2015). Its shell being low, scutes covered with soft skin, fine crystallize olive-green carapace with scattered bright yellow round dashed spots, cream color plastron, grey head and limbs, with bright spots on head and neck as described by (Smith, 1931).

The Indian soft-shelled turtle (Nilssonia gangetica) was identified (Fig. 2.1

B) by flattery surface of jaw raised at its inner marginal edge, head with black lines and no ocelli on carapace of juveniles (Khan, 2015). Most distinct feature was having low carapace dull olive to green, plastron pale, slightly grayish callosities, shorter thancarapace, head broad, massive, dorso-lateral eyes, placed well anterior on snout, lateral ridges on nasal septum (Cuvier, 1825).

The Pangshura smithii (Fig. 2.1 C) was confirmed due to shell bein glow, feeble neural ridge, plastron black, with light boundary (Khan, 2015). According to

Gray, (1863) most notable features this turtle species carapace despressed oval strongly arched highest at midpoint, margin flared spine feeble posterior notched but interiorly shorten plastron. Head is olive with a reddish-brown blotch behind

Table 2.1 Description of sampling sites selected for the study.

Transect Locality (water body) Water quality Major vegetation Water No. type I Samli T.B Sanatorium Water is contaminated with sewage and garbage Cynodon dactylon, Solanum nigrum Running of hospital residual II LoiBher wildlife park Undisturbed area, water is contaminated with sewage Cynodon dactylon,Ziziphusj ujuba Running coming from houses III Junction of Korang and Soan Polluted water with sewage from near by houses Withania somnifera,Cynodon dactylon Running IV Village Bun near Murree Good quality water with stones and rocks are there Parthenium hysterophorus,Carthamu Running soxycantha V Village Bhandar near Sihala Polluted water with sewage coming from houses Datura metal (dhatura), Withania Running of town. somnifera VI Saroba village near Chakari Polluted water due to mixing of sewage from Desmostachya bipinnata (dab) Running vicinity houses VII New Katarian Polluted water due to sewage of factory and Achyranthus aspera (puthkanda) Running houses found nearby VIII Khayaban-e- Sir Syed Polluted water due to mixing of sewage from vicinity Chenopodium ambrosioides(chandan) Running houses IX Rawalpindi Moti Mahal Water bodycontaminated due to sewage of Eichhornia crassipes (water hyacinth) Running vicinity houses and factories polluted water. X Village Sainta Clean and clear, good quality water, stones and rocks Phragmites karka (nar) Running present

XI Village MohraRajgan Contaminated due to the sewage of all adjoining Echinochloa colonum (jungli rice) Running polluted area fall this part of water body XII Village ChohaKhalsa Highly polluted with sewage of surrounding Parthenium hysterophorus (gandi buti) Running residential areas XIII ThohaBahadur Contaminated with sewage of residential areas Acacia modesta (phuali) stagnant XIV JabairPur Least disturbed area, water polluted with garbage Chrysopogon serruletus (grass) stagnant XV Maswal Dam Good quality water Cynodon dactylon (bermuda grass) stagnant XVI KallarKahar Lake More disturbed area, polluted with garbage Typha, Ziziphus jujube and Acacia stagnant modesta (phuali)

Figure 2.1 Topographic map showing location of selected sampling sites in the two districts (Rawalpindi and Chakwal) of the Pothwar Plateau (Downloaded and modified from Google Earth): 1) Loi Bher, 2) Samli TB Sanitorium, 3) Junction of Korang and Sowan Rivers, 4) Bhander, 5) Saroba, 6) New Katarian, 7) Khayaban e Sir Syed, 8) Moti Mahal, 9) Saintha village, 10) Mohra Rajgan, 11) Choha Khalsa, 12) Thoha Bahadur, 13) Jubairpur, 14) Maswal dam, 15) Kallar Kahar Lake, 16) Village Bunn.

A B

C D

Figure 2.2 Photographs of freshwater turtle species recorded in the studyarea

A. The Indian flap shelled turtle (Lissemys punctata)

B. The Indian soft shell turtle (Nilssonia gangetica)

C. The Brown river turtle (Pangshura smithii)

D. The Indian roofed turtle (Pangshura tecta)

eyes, neck and limbs with yellowish brown dots.

The Indian roofed turtle (Pangshura tecta) was identified (Fig. 2.1 D) due to dome-shaped carapace, pastoral surface light, with dark spots with a prominent knobbed neural ridge (Khan, 2015). Arched carapace, more prominent on third vertebral scute, oval with a distinct neural spiked keel, fourth vertebral being longer than wide. Plastron shortening interiorly, carved posterior, pointed snout, posterior half of head with irregular scales, neck dark with narrow yellow streaks (Gray, 1831).

2.4.2 Distribution

Results of the current study showed four freshwater turtle species (Lissemys punctata, Nilssonia gangetica, Pangshura smithi, and Pangshura tecta) were found distributed in various selected water bodies of the district Rawalpindi and two species

(Lissemys punctata and Panshura smithii) in district Chakwal.

2.4.2.1 Indian flap-shelled turtle (Lissemys punctata)

Indian flap shell turtle (Lissemys punctata) were recorded from water bodies of both the districts of Chakwal and Rawalpindi. The species was recorded at all the sampling sites except in village Sainta of district Rawalpindi. Remaining water body of village Sainta was clean and clear with stones and rocks village; which may be unsuitable habitat for this turtle species (Fig. 2.2; Table 2.2). The Lissemys punctata was found distributed in a variety of aquatic habitats in the study area ranging from rivers, shallow streams, salt marshes and stagnant water of ponds and lakes (Fig. 2.2;

Table 2.2).

2.4.2.2 Indian soft shell turtle (Nilssonia gangetica)

The Nilssonia gangetica was recorded at various sampling sites in district

Rawalpindi only was from (Fig. 2.3; Table 2.2) all the sampling sites of district

Chakwal ponds, lakes and dams but probably not suitable habitat for the species. The species prefers running, polluted, muddy and marshy water bodies. In district

Rawalpindi, this turtle species was recorded at 10 out of 12 sampling sites; it was not recorded at village Bun near Murree and Village Sainta, most probably these water bodies were having good water status (Fig 2.2; Table 2.2).

2.4.2.3 Brown river turtle (Pangshura smithii)

The Pangshura smithii exhibited a wide distribution in both Rawalpindi and

Chakwal districts. It was recorded from all the sampling sites except Samli T.B

Sanatorium, Village Bun near Murree, Village Sainta sampling sites in district

Rawalpindi, and Jubair Pur and KallarKahar lake sampling sites in district Chakwal,

(Fig. 2.3, Table 2.2). This turtle is common in river channels and occasionally found in lakes and ponds. It frequently found in muddy water

2.4.2.4 Indian roofed turtle (Pangshura tecta)

Indian roofed turtle was not recorded from any water body samples in the district

Chakwal. However, in district Rawalpindi, it was found distributed at 5 out of 12 sampling sites; Loi Bher Wildlife Park, Junction of Korang and Soan river near judicial colony, Village Bhandar near Sihala, Saroba village near Chakari and Moti Mahal

Rawalpindi, sampling sites were positive regarding distribution of the Indian roofed turtle (Fig 2.5; Table 2.2).

Table 2.2 Distribution of different freshwater turtle species in Rawalpindi and Chakwal districts of the Pothwar Plateau

Transect. Geographical coordinates Locality (water body) L. punctata N. gangetica P. smithii P. tecta No. I 33°51'26.16"N, 73°19'30.218"E Samli T.B Sanatorium + + - - II 33° 34′ 37.857“N73°7' 38.31"E LoiBher wildlife park + + + + III 33°20'13.13", 73°3' 17.94"E Junction of Korang and Soan + + + + river near judicial colony IV 33°33'7.4088"N, 73°14' 41.9676 "E Village Bun near Murree + - - - V 33°33'3.6"N, 73°11'11.4“ E Village bhandar near Sihala + + + + VI 33°18'9.72“ N, 73°46'.13.44"E Soroba village near Chakari + + + + VII 33°38'47.87“N, 73°3'.14.71"E New Katarian + + + - VIII 33°38'19.47“N, 73° 2'.52.7464" E Khayaban-e- Sir Syed + + + - IX 33°36' 34.92“N, 73°3'24.84"E Rawalpindi Moti Mahal + + + + X 33° 28'. 19.56“ N, 73° 29' 22.92 "E Village Sainta - - - - XI 33°28'.25.26“N, 73°24'15.03"E Village Mohra Rajgan + + + - XII 33° 24' 48.24 “N, 73° 28' 32.88 "E Village Choha Khalsa + + + - XIII 32° 56'16.542" N, 72° 42' 10.257"E Thoha Bahadur + - + - XIV 32°55'27.164" N, 72°53'18.491" E Jabair Pur + - - - XV 32° 58'24.85“N, 73°9' 33.48"E Maswal Dam + - + - XVI 32° 46' 25.248“N, 72° 42' 49.96"E Kallar Kahar Lake + - - - *+ sign shows presence of turtle species – shows absence of the turtle species

Figure 2.3 Distribution map of the Indian flap shell turtle (Lissemys punctata) in Rawalpindi and Chakwal districts of the Pothwar Plateau; red dots showing location points where the species was recorded.

Figure 2.4 Distribution map of the Indian soft shell turtle (Nilssonia gangetica) in the Pothwar Plateau; red dots showing location points where this species was recorded.

2.5 DISCUSSION

Reptilian diversity of Pakistan is quite high, the diversity and interesting balance of the herpeto-fauna of the country is probably due to the peculiar zoogeographical position of Pakistan that lies at the transitional zone among

Palearctic, Oriental and Ethiopian zoogeographic region. The protection of the reptiles in the country has received remarkable attention in the recent years. But for the most reptiles as with the majority of other animals, optimum survival depends not on legal protection but as successful functioning of the ecosystem where they live. Freshwater turtle species in tropical and subtropical parts of many Asian countries have been serving as food resource. However, several countries of the region have made few attempts for protection and management of this resource.

The current study focused on freshwater turtles in two districts (Rawalpindi, and Chakwal) of the Pothwar Plateau. The Pothwar Plateau is an important habitat of freshwater turtles, however, detailed ecological studies were lacking. The current study investigated distribution of the freshwater turtle species occurring in the study area. Results revealed in district Chakwal, two species of freshwater turtles were recorded (Lissemys punctata common and Pangshura smithii less common). In district Rawalpindi, four species viz. Lissemys punctata, Nilssonia gangetica, Pangshura smithii and Pangshura tecta, were recorded; Lissemys punctata abundant, Nilssonia gangetica and Pangshura smithii were more common while Pangshura tecta was less common. Akbar et al. (2006) reported 8 freshwater turtles species distributed in different barrages of the Punjab province of Pakistan including P. tecta, P. smithii, Hardella thurjii, Chitra indica, N. gangetica,

Figure 2.5 Distribution map of the Brown River turtle (Pangshura smithii) in water bodies of the two districts (Rawalpindi and Chakwal) of the Pothwar Plateau; red dots showing location points where this species was recorded.

Fig 2.6 Distribution map of the Indian roofed turtle (Pangshura tecta) in the Pothwar Plateau; red dots showing location points where this species was recorded.

Lissemys punctata and Nilssonia hurum; Indian flap shelled turtle (Lissemys

Punctata) being least in number in their study. Similarly, Minton (1996) observed

Indian flap-shelled turtle (Lissemys punctata) as most abundant species, probably because it loves shallow streams, stagnant waters of rivers, marshes, ponds, lakes and often extends in sewage system of metropolitan areas.

Our results have shown that L. punctata has a healthy population in both districts studied and was recorded in about all types of water bodies rivers, streams, canal, ponds, lakes and dams). The second abundant species recorded was P. smithii, which also has a good population in all sampled sites. Mahmood et al.

(2012) investigated the distribution of freshwater turtles in Korang River of

Islamabad Rawalpindi and reported that L. punctata was the most abundant species, N. gangetica less common, while P. smithii was least abundant Minton

(1966) showed that N. gangetica was distributed in Pakistan in the south west part of the country upto Dera Ismail Khan and Sahiwal while Ghalib et al. (1976) reported it from Sindh Indus Valley in Dadu, Hyderabad, Sanghar, Thatta, and

Jacobabad DistrictS, from Punjab, (Multan, Vihari, Muzaffargarh, Lahore, and

Sahiwal districts and the Taunsa Barrage, from Khyber Pakhtunkhwa Mardan, D. I.

Khan,Peshawar Districts and Canal near Charsada) indicating its wider distribution throughout the country.

In the current study L. punctata was distributed in a variety of aquatic habitats ranging from rivers, shallow streams, salt marshes and stagnant water of ponds and lakes. Some earlier studies (Das, 1991; Moll and Moll 2004; Hossain et al., 2008) have shown L. punctata distributed in a variety of aquatic habitats,

ranging from rivers and streams to reservoirs, marshes, ponds, lakes, and even salt marshes, rice fields, gutters, and canals in metropolitan areas.

The Nilssonia gangetica, in the current study was found restricted to rivers, canals and reservoirs where the water was muddy with a turbid bottom generally tending to avoid temporary water bodies. Earlier Khan (2015) indicated that most preferable habitat of the genera N. gangetica and Chitra indica was deep rivers and lakes. The Indian soft-shell turtle also inhabits rivers, lakes, and permanent ponds where it remains buried in the bottom gravel (Das, 1991).

The brown river turtle (Pangshura smithii) has been recorded from muddy water, main river commonly in river channels and occasionally from lakes and ponds. Minton (1966) reported that this turtle species is mainly found in shallow streams, generally found in standing and slow-moving water bodies.

The fourth species, the Pangshura tecta, prefers the aquatic vegetation system, small rivers and stagnant water bodies. This species is mostly confined to emergent vegetation, shallow marginal waters, canals, ponds and puddles. Khan

(2015) showed that Pangshura tecta as confined to all types of stagnant water bodies; domestic ponds, damned ponds (trenches), puddle tanks, old ponds and tributaries of agriculture canals as indicated by Hossain et al. (2006).

2.6 CHAPTER SUMMARY

Pakistan has got extensive range of biodiversity resources including eight species of freshwater turtles, Indian soft-shell turtle, Indian peacock soft shell turtle, Indian narrow-headed soft-shelled turtle, and Indian flap-shell turtle are the

representatives of soft shell species while black spotted turtle, Indian roofed turtle, brown roofed turtle and crowned river turtle are the hard-shelled species. They play important role in biodiversity by cleaning up freshwater resources by scavenging freshwater turtle species in the Pothwar Plateau. The study area included two districts that comprised sixteen selected sampling sites including both lentic and lotic water bodies. Results showed our species in different water bodies of the district Rawalpindi; Indian flap shelled turtle (Lissemys punctata), Indian soft shell turtle (Nilssonia gangetica), Brown river turtle (Panshura smithii) and Indian roofed turtle (Pangshura tecta). The L. punctata was found most abundant, N. gangetica and P. smithii were found common while Pangshura tecta was found least common. However, in district Chakwal, two species of freshwater turtles were recorded; Indian flap shelled turtle (Lessymys punctata) found at Thoha Bahadur,

Jabair Pur, Maswal Dam and Kallar Kahar Lake sampling sites; while Brown river turtle (Panshura smithii) was only found in Thoha Bahdur and Maswal Dam.

Chapter 3

POPULATION ESTIMATION OF FRESHWATER TURTLE

SPECIES IN POTHWAR PLATEAU

3.1 INTRODUCTION

Testudines are the most prosperously developed group of reptiles (Ernst and

Barbour, 1989; Gaffney, 1990). Globally, they consist of 14 families, 319 species and 146 sub-species (Bickham et al., 2007). Turtles existed on the Earth since the rise of the dinosaurs; the first fossil with clear turtle similarity is Odontochelys semitestacea from the Triassic of China (Li et al., 2008; Reisz and Head, 2008), estimated to be 220 million years old, somewhat older than the earliest fossil turtle with a complete shell, Proganochelys quenstedti from the late Triassic of Germany

(Gaffney and Meeker, 1983; Gaffney, 1990; Zug, 1993). Unique turtle shell body and successful body plan have enabled turtles to remain unchanged over some 200 million years of changing climates and despite the evolution of a diverse array of vertebrate predators. Today, tortoises and freshwater turtles are all represented by

460 taxa (species and sub-species) found throughout the tropical and temperate regions of the world (Iverson 1992; Iverson et al., 2003; Fritz and Havas, 2007;

TTWG, 2007; Rhodin et al., 2008).

Turtles contain a familiar group of vertebrates that occupy a broad range of habitats and ecological functions. Some species are important in energy flow, and may dominate vertebrate biomass, particularly in aquatic habitats (Congdon et al.

1986).

Freshwater turtles generally spend majority of their time in aquatic habitats, while many species travel between wetlands (Bowne, 2008; Roe and Georges,

2008) and engage travel overland to upland nesting areas (Carr, 1952; Congdon et al., 1987). The use of both aquatic and upland habitat by freshwater turtles indicates that conservation of wetland habitat alone is inadequate to conserve many populations of freshwater turtles. While nesting-related travel accounts for only a small percentage of female turtle’s annual time budget, the risk of road mortality

(Haxton, 2000; Steen et al., 2006) and human exploitation (Suganuma et al., 1999) increases dramatically during nesting forays, and high mortality of adult females during nesting-related travel may skew population sex ratios (Steen and Gibbs,

2004). Because road mortality is experienced only when turtles are traveling between more broadly used habitat patches, the problem involves both landscape structure and connectivity. The often-extensive travel by freshwater turtles through a terrestrial environment (Bowne, 2008) and use of multiple wetland covers throughout the year (Joyal et al., 2001), suggests that effective protection strategies should be undertaken at the landscape level.

Freshwater turtles live in marshy and wetland habitats, many of which have vanished or been considerably modified in the last 100 years. Human distraction includes habitat alteration, water pollution, terrestrial alteration, channelization, and sand mining, all of which have a negative effect on freshwater turtles throughout the world (Moll and Moll, 2004).

Population data on freshwater turtles is required for management and conservation purposes; however, such information is scanty (Gibbons et al., 2000).

Few studies have been conducted so far on Indian freshwater turtle’s taxonomy and their broad distributional ranges (Smith 1933; Pritchard, 1979; Daniel, 1983; Moll,

1984; Das, 1985). Population dynamics, biology and ecology of freshwater turtles are deemed as one of the presenting problem to device management policies (Moll,

1984).

Turtles are considered to be of distinct protection concern because they have low reproductive output, late maturity and habitat requirement of both wetlands and terrestrial environments (Congdom and Gibbons, 1996; Klemens,

2000). The population composition has salient ecological and conservation implications and can fundamentally affect the structure of communities. For example, sudden change in sex ratios can result in smaller effective population size and can thus lower recruitment and also alter the age composition of the population

(Primack 1998; Smith and Smith, 2001).

The populations of different species of turtles have declined in the recent past. Gibbons et al. (2000) showed that protection disasters are the main reason of decreased population of reptiles all over the world. Moreover, recently habitat loss and fragmentation have also been focused (Curtin, 1997; Sarre, 1998; Gibbons et al., 2000). Similarly, humiliation of habitat and displacement of wetlands may also result in abnormal population structure or population decline and disappearance

(Dodd, 1990; Reese and Welsh, 1996). Loss of aquatic vegetation has also contributed to decrease in turtle populations. The mortification of water ways is another great threat to the viability of freshwater turtles (Cann, 1993). Moreover, construction of dams, water level variation, channelization and water depletion for

irrigation have also badly affected wetland habitats resulting in the loss of biodiversity.

Freshwater turtles are a group of high conservation concern, with about 40

% of the world’s turtle species currently listed as globally “threatened” by IUCN

(Rhodin et al. 2010). The decline of many freshwater turtle species is attributed to habitat loss and fragmentation (Turtle Conservation Fund 2002). Exploitation and uncontrolled trade are the primary causes for acute decline in many turtle species, especially those from Asia, due to some anthropogenic threats; habitat loss and degradation also being major factors in widespread declines (van Dijk et al., 2000,

Gibbons et al., 2000, Turtle Conservation Fund, 2002). The position of Asian chelonians is put into risk by habitat destruction (Collins, 1990). Reproductive rate of most species considered low, decreased drastically from natural habitat (Sandra and Daniela, 2000). The persistence of such an ancient and iconic group is under concerted assault, and turtles have become prominent victim of global biodiversity crisis. Without directed, calculated conservation planning, a significant portion of turtle diversity could be lost over the next century (van Dijk et al., 2000).

Many freshwater turtle populations are found in aquatic habitats under various forms of pressure from human activities (Ban-croft et al., 1983; Cagle,

1942; Gibbons, 1970; Moll, 1980; Parker, 1984), and many of these are located in areas of urban development. Because of anthropogenic effects, populations located in urban areas should exhibit variation in aspects of their population ecology and life history characteristics when compared to populations in nonurban areas.

3.2 REVIEW OF LITERATURE

A total of 167 reptile species have been recorded so far from Pakistan

(Auffenberg and Khan, 1991; Ghalib et al., 1976; Rehman and Iffat, 1997). From the Sindh province of the country, about 105 reptile species have been reported

(Khan et al., 2012).

Freshwater resources of Pakistan are dominated by the Indus River system, which serves as drainage basin for Himalayas. Most of wetlands support the inhabitant the needs of growing population, the humiliation of freshwater ecosystem by a variety of aggravation have increased exponentially. As a result, many ecosystems are in need of some drastic corrective restoration (Khan et al.,

2015).

Different methods are used to estimate the population of turtles species in different countries but in Pakistan applied “direct observations” method on basking and floating turtles in the river Indus where by they reporteds 6 species of turtles including. Pangshura smithi, Kachuga tecta, Hardella thurgi, Chitra indica,

Nilssonia gangetica and Lissemys punctata from the study area. Chitra indica and

Pangshura smithii were found to abundant Azam et al. (2005). Some information was updated by Khan (2015) about freshwater turtles fauna of Indus Valley of

Pakistan; he reported up river turtles belonging to two families; the pond turtle family “Emydidae” that includes the genera Geoclemys, Hardella and Pangshura and the softshell family “Trionychidae” that includes the genera Nilssonia, Chitra and Lissemys punctata.

Akbar et al. (2006) applied different methods for population estimation; direct count, basking method during severe cold weather conditions and baited

hook method. He reported approximately a total of 3528 turtles from 19 barrages of the Punjab province and belonging eight species and two families viz.,

Geoemydidae including the species Brown River turtle (Pangshura smithii), Indian

Saw-backed turtle (Pangshura tecta), Brahminy Rrive turtle (Hardella thurgi),Spotted Pond turtle (Geoclemys hamiltonii) and the family Trionychidae including species Indian Soft-shell turtle (Nilssonia gangetica), Peacock Soft-shell turtle (Nilssonia hurum), Narrow-headed Soft-shell turtle (Chitra indica), and

Indian Flap-shell turtle (Lissemys punctata) in Punjab province by using direct observations and drag net methods.

Safi and Khan (2014), slected three rivers (Jindi, Swat and Kabul) of district Charsadda (Khyber Pakhtunkhwa) district Charsadda to find out the

Population status of freshwater turtle species. Four methods were applied viz; direct count, baited hook, basking during sever weather condition and interviews were couducted. They reported two families of freshwater turtles; “Geoemydidae” including crowned river turtle (Hardella thurjii), Brown roofed turtle (Pangshura smithii) and Indian roofed turtle (Pangshura tecta) and the other family

“Trionychidae” Indian narrow-headed soft-shell turtle (Chitra indica), Indian soft- shell turtle (Nilssonia gangetica), Indian peacock soft-shell turtle (Nilssonia hurum) and Indian Flap-shell turtle (Lissemys punctata) Safi and Khan (2014).

Similarly, two families of freshwater turtles have been reported from the rivers of the Punjab province, Pakistan by Brooks et al. (1988) and Lovich, (1994).

Mahmood et al. (2012) used “visual observation” with the help of a binocular (Olympus 10Xx 50X) by walking along each transect and “cast net”

method for relative abundance of freshwater turtle species from Korang River,

Islamabad/Rawalpindi. They recorded three species of freshwater turtles viz. Indian flap shelled turtle Lissemys punctata, Indian soft shelled turtle (Nilssonia gangetica) and brown river turtle Pangshura smithii. Lissemys puncta was found most abundant species are Nilssonia gangetica and Pangshura smithii were less common. Various researchers (Azam and Saeed, 2011; Noureen, 2007; Noureen et al., 2012; Safi and Khan, 2014) have studied abundance of freshwater turtle species in Pakistan. The main findings include 02 families, 06 genera and 08 species of freshwater turtles. The family Geoemydidae consists of hard-shelled turtles viz.

Spotted Pond turtle (Geoclemys hamiltonii), Crowned river turtle (Hardella thurjii), Brown roofed turtle (Pangshura smithii), and Indian roofed turtle

(Pangshura tectum). The second family, Trionychidae, comprises of softshell turtles viz. Indian narrow-headed soft-shell turtle (Chitra indica), Indian soft-shell turtle (Nilssonia hurum) and Indian flapshelled turtle (Lissemys punctata and ersonii).

In 1990, thefreshwater turtle trade was first highlighted in Pakistan; an analytical study was done by WWF-Pakistan and the Sindh Wildlife Department for TRAFFIC International in 1996. Illegal turtle trade, especially soft-shell species body parts for food and traditional medicine were reported by Azam (2006). In

Khyber Pakhtunkhwa, illegal trade of turtles and their body parts was highlighted in the year 2006 (Noureen et al., 2012).

Lissemys punctata, Nilssonia gangetica and Nilssonia hurum (soft-shell turtles) are being utilized in low quantity in Pakistan. In India, record of the captive

stocks of turtles in zoos was prepared based on questionnaire surveys, response was received from 35 zoos, the listed species in zoos were Lissemys punctata and

Indian star tortoise (Geochelone elegans) covering all but two biogeography zones.

Remarkable endangered species such as Nilssonia hurum, Chitra indica, Cyclemys dentate, Melanochelys tricarinata. Also a new status listing was recommended for turtles and tortoises to include in India (Wildlife Protection Act, 1972). One of the freshwater turtle species (Pangshura smithii) was categorizes as “Endangered” in the Indian Wildlife Protection Act 1972 and also enlisted in IUCN Red Book (Rao,

2006).

Turtle populations are being eliminated rapidly not being protected or conserved seriously by any Government department in Pakistan; therefore, some species populations are becoming threatened (Khan, 2015). In 1976, Pakistan became a signatory to the Convention on International Trade in Endangered

Species of fauna and flora (CITES). In May 2012, Government of Pakistan approved the Federal CITES Law, for effective enforcement of CITES legislations in Pakistan. There are many anthropogenic threats being faced by the freshwater turtles, due to which conservation was needed. Unfortunately, freshwater turtles were not earlier on included in the list of protected animals in all provincial

Wildlife Protection Acts in Pakistan. On 22 August 2007, Khyber Pakhtunkhwa

Wildlife Department revised the Wildlife Protection Act, 1975 and all the freshwater turtles of the Province were included in the list of protected animals.

After that Punjab Wildlife and Parks Department revised the Punjab Wildlife

Protection Act 1974, on November 30 2007, and made an amendment to include the Order Chelonia (Turtles and Tortoises) in the list of protected animals

(Schedule III) (Anonymous, 2005).

3.3 MATERIALS AND METHODS

3.3.1 Study Design

The current study was carried out at 16 water bodies of the study area; a total of sixteen sampling water bodies were visited from March 2012 to Feburary

2014 on monthly basis for data collection about population estimates of the four species of the freshwater turtle using two different methods viz. “Capture Mark and

Recapture” and “Direct count” methods.

3.3.2 Capture, Mark and Recapture Method (CMR)

A cast net was operated at each selected water body to catch the turles; the cast net was made up of nylon, having150 cm radius with a mesh size of 5 cm. Ten

(10) attempts were made with cast net at each site during each visit to capture turtles (Fig. 3.1). During this process, firstly, “bait” was cast at specific sites to attract turtles so that successful netting could be made possible. It was assumed that all turtle species were equally attracted to the bait used. The bait used consisted of poultry intestines and liver of chick, and goat Sutherland (1998). The physical characteristics of the captured turtles like body weight, carapace length and width, plastron length and width, were recorded. The captured turtles were counted species-wise and marked by notching in their marginal scutes (Gibbons, 1976) and released back into the same water body after data recording (Fig. 3.2 A).

The study area was visited each month during the study period. Each time animals were captured by using cast net. The captured animals were marked by

notching in the marginal scutes and again released unharmed back into the environment (Fig. 3.2 B). On the next visit, the turtles were again captured from the same site with the help of cast net and counted for marked and unmarked ones.

The unmarked turtles were marked by notching in the marginalscutes and again released back into the population. In this way on each visit it was expected that the number of unmarked animals were going decrease. The dates of capture of the turtles were recorded and at the end of the study period, the captured histories were mathematically analyzed to estimate the population size and survival by using the formula;

N= (M+1) (C+1) – 1

R+1

Where; N is the Estimate of total population size, M is the total numbers of turtles captured and marked on the first visit, C is the total number of turtles captured on the second visit, and R is the numbers of turtles captured on the first visit that were then recaptured on the second visit.

3.3.3 Direct Count Method

At each selected site, visual observations were made by naked eye and also by using Nikon binocular (10 X 16 mm) to count number of turtles as possible with in a circular central zone along the wetland, such as lake, pond, water reservoirs, and by walking for one kilometer transect along a river or stream with 250 meters width on either side (Fig. 3.3). The turtle species observed were identified and counted on distinct morphological features like color of carapace, and other features following Burnham et al. (1980). The population of each species was then

Figure 3.1 Acast net is being cast at Saroba village near Chakari which is associated with soan river.

A B

Figure 3.2 Photograph of Pangshura smithii showing marginal scute notching process;

A) Notching process for marking purpose, B) a notched specimen.

Figure 3.3 Taking visual observations of freshwater turtles at Village Bhandar site near Sihala district Rawalpindi.

estimated by using following formula:

P = AZ 2YX

Where, P is the Population, A is the total area, Z is the numbers observed, Y is the average flushing distance, and X is the length of strip.

3.3.4 Statistical Analysis

The data about turtles populations were presented in the form of graph prepared in Microsoft excel 2013, the populations of fresh water turtlespecies at different localities, and the two methods used (CMR and direct count methods) were compared through Two-way Analysis of Variance (ANOVA). The significant results of ANOVA were later on processed through Post Hoc Test. p value less than 0.05 was considered as significant.

3.4 RESULTS

3.4.1 Population Estimates of Freshwater Turtles District Rawalpindi

Two methods (CMR and Direct Count) were applied for estimating the populations of various turtle species in district Rawalpindi during the study period.

3.4.1.1 Capture, Mark and Recapture Method (Relative abundance of turtles)

Using Capture, Mark and Recapture (CMR) method, in district Rawalpindi, twelve (N=12) sampling sites or water bodies were sampled for estimating populations of different turtle species. A total of 1681 individuals of turtles belonging to two families (Trionychidae; soft-shell turtles) and Geoemydidae;

Table 3.1 Relative abundance of four species of the freshwater turtles live captured from twelve (N=12) different sampling sites of district Rawalpindi using Capture, Mark, and Recapture (CMR) method from 2012 to2014(total time spent at each sampling site was 3 hours, numbers of attempts made by using cast net =10 on 1Km transect during all 24 visits). Parentheses indicate % population of the species at that particular site.

Soft -shell turtle Hard- shell turtle Sr. No. Selected sites L. punctata D/Km² N. gangetica D/Km² P. smithii D/Km² P. tecta D/Km² Total % 1. Samli T.B Sanatorium 43(6.3) 3.5 17.6 (4.0) 1.4 - - - - 60.6 3.5 2. LoiBher wildlife park 77.1(11.3) 6.4 38.5 (8.7) 3.2 56.9 4.7 15.8 1.3 188.3 11.1 (13.3) (11.6) 3. Junction of Korang and 80.8 (11.8) 6.7 59.4 (13.5) 4.9 69.7 5.8 32.8 2.7 242.7 14.4 Soan river, near judicial (16.3) (24.2) colony 4. Village Bun near Murree 37.9 (5.5) 3.1 ------37.9 2.2 5. Village Bhandar near 64.2 (9.4) 5.3 60.5 (13.7) 5.0 36.5 (8.5) 3.0 30.8 2.5 192 11.4 Sihala (22.7) 6. Soroba village near 68.5 (10.0) 5.7 77.4 (17.6) 6.4 42.8 3.5 29.2 2.4 217.9 12.9 Chakari (10.0) (21.5) 7. New Katarian 62.3 (9.1) 5.1 53.5 (12.1) 4.4 42.6 (9.9) 3.5 - - 158.4 9.3 8. Khayaban-e-Sir Syed 91.2 (13.4) 7.6 52.7 (11.9) 4.3 65.2 5.4 - - 209.1 12.4 (15.2) 9. Rawalpindi MotiMahal 61.4 (9.0) 5.1 36.8 (8.3) 3.0 50.2 4.1 26.6 2.2 175 10.4 (11.7) (19.6)

10. Village Sainta ------11. Village MohraRajgan 45.6 (6.7) 3.8 21.9 (4.9) 1.8 29.9 (7.0) 2.4 - - 97.4 5.7 12. Village ChohaKhalsa 47.9 (7.0) 3.9 21.1 (4.8) 1.7 33.2 (7.7) 2.7 - - 102.2 6.0 Total 679.9 56.2 439.4 36.1 427 35.1 135.2 11.1 1681.5 Mean± S.E 61.8±4.9 5.1±0.4 43.9±5.7 3.6±0.4 49.2±4.0 3.9±0.3 27.0±1.9 2.2±0.1 Relative abundance 40.4% 26.1% 25.3% 8.0%

Frequency (F)F=NT/TT 11/12=0.9 10/12=0.8 9/10=0.8 5/12=0.4 *NT= No. of transects in which turtles were observed, TT= Total no. of transect, F= Frequency=NT/TT, D= Density /Km². *Brackets denote % population of the species at that particular site. *43 is total number of turtles captured from Samli T.B Sanatorium site while 6.3 is relative percentage (%) from sixteen sites.

hard-shell turtles) and four different species were recorded from March 2012 to

February 2014. The most abundant species was Lissemys punctata (40.4%), followed by Nilssonia gangetica (26.1%) and Pangshura smithii (25.3%) whereas least abundant species was Pangshura tecta (8.0%) (Table3.1). These turtle species were recorded in good numbers (N=242) at the “junction of Korang and Soan rivers site near Judicial Colony (Rawalpindi), while least (N=37) were recorded at the “Village Bun” near Murree.

3.4.1.1.1 Frequency (F) and density (D)

Among all four species of freshwater turtles recorded, Lissemys punctata was found the most frequent species (0.9) in district Rawalpindi during the current study period by Capture, Mark and Recapture (CMR) method, followed by

Nilssonia gangetica and Pangshura smithii (0.8 each), while the least frequent species (0.4) was Pangshura tecta (Table 3.1).

Similarly, Lissemys punctata showed highest average density (5.1±0.4/km²) at all selected sampling sites, followed by Nilssonia gangetica and Pangshura smithii (3.6±0.4/km²each) and least (2.2±0.1/km²) by Pangshura tecta (Table 3.1).

3.4.1.1.2 Seasonal variation CMR method

By CMR method during summer/ spring from 2012 to 2014, a total N=1515 freshwater turtles were recorded from all sampling sites of the district Rawalpindi;

L. punctata represented 39.3%, N. gangetica 27.0%, P. smithii 26.1% and P. tecta

8.0% (Table 3.2). Maximum population (218.2) of freshwater turtles was recorded at the “Junction of Korang and Soan river” site and minimum (29.9) at “Village

Table 3.2 Relative abundance of four species of the freshwater turtles live captured from twelve (N=12) different sampling sites of district Rawalpindi using

Capture, Mark, and Recapture (CMR) method during summer/spring (total time spent at each sampling site was 3 hours, numbers of attempts made by using cast net =10 on 1Km transect during all 24 visits).

Sr. No. Selected sites Soft -shell turtle Hard- shell turtle L. D/Km² N. D/Km² P. D/Km² P. tecta D/Km² Total % punctata gangetica smithii 1. Samli T.B Sanatorium 37.2(6.2) 3.1 15.4 (3.7) 1.2 - - - - 52.6 3.4 2. Loi Bher wildlife park 75.3(12.6) 6.2 34 (8.3) 2.8 54.8 4.5 15.8 1.3 179.9 11.8 (13.8) (13.8) 3. Junction of Korang 69.3 (11.6) 5.7 56.9 (13.9) 4.7 62 (15.6) 5.1 30(26.3) 2.5 218.2 14.3 and Soan river 4. Village Bun near 29.9 (5.0) 2.4 ------29.9 1.9 Murree 5. Village Bhandar near 62 (10.3) 5.1 53 (12.9) 4.4 34.2 2.8 24 (21.0) 2.0 173.2 11.4 Sihala (8.6) 6. Soroba village near 62.4 (10.4) 5.2 71.8 (17.5) 5.9 35.1 2.9 23.9 1.9 193.2 12.7 Chakari (8.8) (20.9) 7. New Katarian 53.4 (8.9) 4.4 52.3 (12.7) 4.3 40.9 3.4 - - 146.6 9.6 (10.3) 8. Khayaban-e-Sir Syed 68.8 (11.5) 5.7 51.2 (12.5) 4.2 58 (14.6) 4.8 - - 178 11.7 9. Rawalpindi 52.4 (8.7) 4.3 31.7 (7.7) 2.6 50.3 4.1 20.3 1.6 154.7 10.2 MotiMahal (12.6) (17.8) 10. Village Sainta ------11. Village MohraRajgan 41.2 (6.9) 3.4 21.9 (5.3) 1.8 27.6 2.3 - - 90.7 5.9 (6.9) 12. Village ChohaKhalsa 44.3 (7.4) 3.6 21.1 (5.1) 1.7 33.4 2.7 - - 98.8 6.5 (8.4)

Total 596.2 49.1 409.3 33.6 396.3 32.6 114 9.3 1515.8 Mean± S.E 49.6±6.0 4.4±0.3 34.1±6.7 3.3±0.4 33.0±6.5 3.6±0.2 9.5±3.5 1.8±0.1 Relative 39.3% 27.0% 26.1% 8.0% abundance Frequency F=NT/TT 11/12=0.9 10/12=0.8 9/10=0.8 5/12=0.4 (F) * NT= No. of transects in which turtles were observed, TT= Total no. of transect, F= Frequency=NT/TT, D- Density /Km². *Brackets denote % population of the species at that particular site. *37.2 is total number of turtles captured from Samli T.B Sanatorium site while 6.2 is relative percentage (%) of sixteen sites.

Table 3.3 Relative abundance of four species of the freshwater turtles live captured from twelve (N=12) different sampling sites of district Rawalpindi using Capture, Mark, and Recapture (CMR) method during winter/fall (total time spent at each sampling site was 3 hours, numbers of attempts made by using cast net =10 on 1Km transect during all 24 visits).

Sr. No. Selected sites Soft -shell turtle Hard- shell turtle L. punctata D/Km² N. gangetica D/Km² P. smithii D/Km² P. tecta D/Km² Total %

1. Samli T.B Sanatorium 5.7(6.2) 0.4 2.3 (7.6) 0.1 - - - - 8.0 4.2 2. LoiBher wildlife park 6.8(7.4) 0.5 5(16.7) 0.4 4.8 (9.7) 0.4 - - 16.6 8.7 3. Junction of Korang and Soan 9.8 (10.7) 0.8 3.4 (11.3) 0.2 8.8 (17.9) 0.7 2.3(11.2) 0.1 24.3 12.7 river, near judicial colony

4. Village Bun near Murree 8 (8.7) 0.6 ------8.0 4.2 5. Village Bhandar near Sihala 8 (8.7) 0.6 7.2(24.0) 0.6 5.8 (11.8) 0.3 7.8 (38.2) 0.6 28.8 15.1

6. Soroba village near Chakari 7 (7.6) 0.5 6.8 (22.7) 0.5 7.5 (15.3) 0.6 6.8(33.3) 0.5 28.1 14.7

7. New Katarian 10.3 (11.3) 0.8 - - 6 (12.2) 0.5 - - 16.3 8.5

8. Khayaban-e-Sir Syed 21.4 (23.5) 1.7 2.7 (9.0) 0.5 8 (16.3) 0.6 - - 32.1 16.8

9. Rawalpindi MotiMahal 8 (8.7) 0.6 2.5 (8.3) 0.2 3.5 (7.1) 0.2 3.5 (17.1) 0.2 17.5 9.1

10. Village Sainta ------11. Village MohraRajgan 6 (6.5) 0.5 - - 4.6 (9.3) 0.3 - - 10.6 5.5 12. Village ChohaKhalsa ------Total 91 7.0 29.9 2.5 49 3.6 20.4 1.4 190.3 Mean± S.E 8.2±1.4 0.7±0.1 2.9±0.7 0.4±0.0 5.4±0.7 0.4±0.0 4.0±0.9 1.8±0.1 Relative 47.8% 15.70% 25.7% 10.7% abundance Frequency F=NT/TT 10/12=0.8 7/12=0.5 8/10=0.6 4/12=0.3 (F) * NT= No. of transects in which turtles were observed, TT= Total no. of transect, F= Frequency=NT/TT, D- Density /Km². *Brackets denote % population of the species at that particular site.

Figure 3.4 Graphical representation of seasonal variation in populations of freshwater turtles in district Rawalpindi using CMR method.

Figure 3.5 Seasonal variation in populations of freshwater turtles in district

Rawalpindi using direct count method.

Table 3.4 Relative abundance of four species of the freshwater turtles from twelve (N=12) different sampling sites of the district Rawalpindi using

“direct count” method from 2012 to 2014.

Sr. No Soft –shell turtle Hard- shell turtle

Selected sites L. D/Km² N. D/Km² P. smithii D/Km² P. tecta D/Km² Total % punctata gangetica 1. Samli T.B Sanatorium 74 (8.7) 6.1 41( 7.1) 3.4 - - - - 115 4.6 2. LoiBher wildlife park 107 (12.6) 8.9 48 (8.3) 4 97 (12.8) 8.0 30 (11.6) 2.5 282 11.4 3. Junction of Korang and 7.9 7.7 5.2 Soan river near judicial 4.1 colony 95 (11.2) 50 (8.6) 93(12.3) 63 (24.4) 301 12.1 4. Village Bun near Murree 75 (8.8) 6.2 ------75 3.0 5. Village Bhandar near Sihala 96 (11.3) 8.0 76 (13.2) 6.3 127 (16.8) 10.5 62 (24.4) 5.1 361 14.6 6. Saroba village near Chakari 89 (10.5) 7.4 86 (14.9) 7.1 129 (17.1) 10.7 53 (20.5) 4.4 357 14.4 7. New Kataria 78 (9.2) 6.5 65 (11.3) 5.4 113 (15.0) 9.4 - - 256 8.1 8. Khayaban-e- Sir Syed 92 (10.9) 7.6 70 (12.1) 5.8 71(9.4) 5.9 - - 233 10.3 9. Rawalpindi MotiMahal 76 (9.0) 6.3 70 (12.1) 5.8 45 (5.9) 3.7 50 (19.3) 4.1 241 9.7 10. Village Sainta ------0 0 11. Village MohraRajgan 47 (5.5) 3.9 40 (6.9) 3.3 44 (5.8) 3.6 - - 131 5.3 12. Village ChohaKhalsa 55 (6.5) 4.5 29 (5.0) 2.4 34 (4.5) 2.8 - - 118 4.7 Total 844 73.3 575 47.6 753 62.3 258 21.3 2470

Mean± S.E 73.6±8.3 6.6±0.4 47.9± 8.0 4.7±0.4 62.7± 14.1 7.4±0.8 21.5± 8.0 7.1±0.3

Relative abundance 34.1 % 23.2% 30.4% 10.4% Frequency (F) 11/12=0.9 10/12=0.8 9/12= 0.7 5/12= 0.4

Bun near Murree” site (Table 3.2; Fig. 3.3).

During fall/winter season, total numbers of freshwater turtles recorded at all selected water bodies of the district Rawalpindi from 2012 to 2014 were just 190

(as compared to 1515 during summer season); however, L. punctata represented

47.8%, Nilssonia gangetica 15.70%, Pangshura smithii 25.7% and P. tecta 10.7%

(Table 3.3). Maximum population of turtles (32.1) during this season was recorded at “Khayaban-e-Sir Syed while minimum (zero) at “village Sainta” site (Table 3.3;

Fig. 3.4).

3.4.1.2 Direct Count Method

3.4.1.2.1 Abundance of freshwater turtles

In district Rawalpindi, the same twelve (N=12) sites or water bodies were sampled for estimating populations of different turtle species (Table 3.4) using direct count method. Results were quite similar to those by CMR method. Total numbers of freshwater turtles recorded from all sampling sites of this district were

2470; L. punctata was the most abundant species (34.1%), followed by P. smithii

(30.4 %), N. gangetica (23.2 %), and least (10.4 %) was found P. tecta (Table 3.4).

Highest population of turtles was recorded at village Bhandar near Sihala sampling site (14.6 %) while minimum at village Bun near Murree (3.0%).

3.4.1.2.2 Frequency (F) and density (D) of turtles

The direct count method used also showed L. punctata (0.9) as most frequent species in the district Rawalpindi during the study period, followed by

Nilssonia gangetica (0.8), Pangshura smithii (0.7) and least frequent (0.4) was

Table 3.5 Relative abundance of four species of the freshwater turtles from twelve (N=12) different sampling sites of district Rawalpindi using direct count method during summer/spring season.

Sr. No Soft –shell turtle Hard- shell turtle

Selected sites L. punctata D/Km² N. gangetica D/Km² P. smithii D/Km² P. tecta D/Km² Total % 1. Samli T.B Sanatorium 67 (9.0) 5.5 38 (7.3) 3.1 - - - - 105 4.9 2. LoiBher wildlife park 87 (11.6) 7.2 43 (8.7) 3.5 79 (12.2) 6.5 28 (12.1) 2.3 237 11.2 3. Junction of Korang and Soan river near judicial colony 80(10.7) 6.6 41(8.3) 3.4 87 (13.4) 7.2 52 (22.6) 5.2 260 12.3 4. Village Bun near Murree - - - 63(8.4) 5.2 - - - 63 2.9 5. Village Bhandar near Sihala 80 (10.7) 6.6 60 (12.1) 5 109(16.8) 9.0 52 (22.6) 4.3 301 14.2 6. Saroba village near 6.0 Chakari 73(9.8) 82 (16.6) 6.8 108(16.7) 9.0 49 (21.3) 4.0 312 14.7 7. New Kataria 64 (8.6) 5.3 60 (12.1) 5 92 (14.2) 7.6 - - 216 10.2 8. Khayaban-e- Sir Syed 75 (10.0) 6.2 56 (11.3) 4.6 64 (9.9) 5.3 - - 195 9.2 9. Rawalpindi MotiMahal 5.5 67 (9.0) 58 (11.7) 4.8 41 (6.3) 3.4 49 (21.3) 4.0 215 10.1 10. Village Sainta ------0 11. Village MohraRajgan 40 (5.3) 3.3 32(6.4) 2.6 35(5.4) 2.9 - - 107 5.0 12. Village ChohaKhalsa 48 (6.4) 4.0 23(4.6) 1.9 30 (4.6) 2.5 - - 101 4.7 Total 744 61.4 493 40.7 645 53.4 230 19.8 2112 100

Mean± S.E 67.6±4.0 10.2±0.3 50.5±5.1 7.4±0.4 71.6±8.8 10.6±0.7 46±2.9 6.6±0.3 4.9 Relative abundance 35.2 % 23.3% 30.5% 10.8% Frequency (F) 11/12=0.9 10/12=0.8 9/12= 0.7 5/12= 0.4

Table 3.6 Relative abundance of four species of the freshwater turtles from twelve (N=12) different sampling sites of district Rawalpindi using direct count method during winter/fall season.

Sr. No Soft –shell turtle Hard- shell turtle

Selected sites L. punctata D/Km² N. gangetica D/Km² P. smithii D/Km² P. tecta D/Km² Total % 1. Samli T.B Sanatorium 7(5) 0.5 3(3.7) 0.2 - - - - 10 2.7 2. LoiBher wildlife park 1.6 0.4 1.5 0.1 20(14.2) 5(6.1) 18(15.9) 2(7.1) 45 12.4 3. Junction of Korang and Soan river near judicial colony 1.2 0.7 0.5 0.9 15(10.7) 9 (11.1) 6(5.3) 11 (39.2) 41 11.3 4. Village Bun near Murree - - - 12(8.5) 1.0 - - - 12 3.3 5. Village Bhandar near Sihala 1.3 1.3 1.5 0.8 16 (11.4) 16 (19.7) 18 (15.9) 10(35.7) 60 16.5 6. Saroba village near Chakari 1.3 0.3 1.7 0.3 16(11.4) 4(4.9) 21(18.5) 4(14.2) 45 12.4 7. New Kataria 14(10) 1.1 5(6.1) 0.4 21(18.5) 1.7 - - 40 10.4 8. Khayaban-e- Sir Syed 17(12.1) 1.4 14(17.2) 1.1 7(6.1) 0.5 - - 38 10.4 9. Rawalpindi MotiMahal 0.7 1.0 0.3 0.0 9(6.4) 12(14.8) 4(3.5) 1(3.5) 26 7.1 10. Village Sainta ------0 0 11. Village MohraRajgan 7(5) 0.5 7(8.6) 0.5 9(7.9) 0.7 - - 23 6.3 12. Village ChohaKhalsa 7(5) 0.5 6(7.4) 0.5 9(7.9) 0.7 - - 22 6.0 Total 140 11.1 81 6.4 113 9.1 28 2.1 362 Mean± S.E 12.7± 1.3 1.8±0.1 8.1± 1.2 1.1±0.1 12.5± 1.9 1.8±0.1 5.6± 1.3 0.7±0.1 Relative abundance 38.6 % 22.3% 31.2% 7.7% Frequency (F) 11/12=0.9 10/12=0.8 9/12= 0.7 5/12= 0.4

Figure 3.6 Seasonal variation in populations of freshwater turtles from district

Chakwal using CMR method during the study period.

Figure 3.7 Seasonal variation in populations of freshwater turtles from district

Chakwal using direct count method during the study period.

Pangshura smithii (Table 3.4).

Similarly, the density estimates of the four turtle species from selected sites of the district by direct count method revealed highest average density of

6.6±0.4/km² for Lissemys punctata, followed by 4.7±0.4/km² for Pangshura smithii

7.4±0.8/ Km² for Nilssonia gangetica and least 7.1±0.3 Km² for Pangshura tecta

(Table 3.4).

3.4.1.2.3 Seasonal variation in turtle populations by direct count method

During summer/spring season (from 2012 to 2014), a much greater number of (N=2112) freshwater turtles were recorded from twelve selected sites of the district using direct count method; L. punctata represented 35.2 %, P. smithii

30.5%, N. gangetica 23.3 % and P. tecta 10.8%. Maximum numbers of freshwater turtles (312) were recorded at Saroba village near Chakari and minimum (63) village Bun near Murree (Table 3.5).

However, during winter/fall season, numbers of freshwater turtles recorded was very low (N 362, as compared to 2112 during summer season); Lissemys punctata represented 38.6 %, Nilssonia gangetica 22.3 %, Pangshura smithii 31.2

% and Pangshura tecta 7.7 %. Maximum numbers of turtles (60) were recorded at

“Village Bhandar” near Sihala while minimum (zero) at “village Sainta” site (Table

3.6; Fig. 3.5).

3.4.2 Population Estimates of Freshwater Turtles in District Chakwal

Two methods (CMR and Direct count method) were applied for estimating the populations of turtle species in district Chakwal during the study period.

3.4.2.1 Capture, mark and recapture method-turtles abundance

Using capture, mark and mecapture (CMR) method, in district Chakwal, four (N=4) sampling sites or water bodies were sampled for estimating populations of different turtle species. A total of 483 turtles belonging to two different species were recorded from March 2012 to February 2014; Lissemys punctata was found most abundant (75.1%) whereas the relative abundance of Pangshura smithii was recorded to be 24.9% (Table 3.7). Maximum numbers (N=251) of freshwater turtles (both species) were recorded at the “Thoha Bahadur” sampling site while least (N=59) were recorded at the “Kallar Kahar Lake” sampling site.

3.4.2.2Frequency (F) and density (D)

At four sampling sites of the district Chakwal, Lissemys punctata was found the most frequent species (1), by Capture Mark and Recapture (CMR) method, while less frequent was P. smithii (0.5). Similarly, Lissemys punctata showed highest average density (7.5±2.5/km²) while Pangshura smithii showed an average density of 0.9±1.8/km² (Table 3.7).

3.4.2.3 Seasonal variation in turtle populations (by CMR method)

During summer/spring season from 2012 to 2014, a total N= 349 freshwater turtles were recorded from all foursampling sites of the district Chakwal; Lissemys punctata represented 76.5% and Pangshura smithii 23.4%. Maximum population

(132) of turtles was recorded at the “Thoha Bahadur” site and minimum (47) at

“Kallar Kahar Lake” site (Table 3.8).

Table 3.7 Relative abundance of freshwater turtle species from four selected sampling sites of district Chakwal during 2012-2014 by using CMR method.

Soft- shell turtle Hard- shell turtle

Sr. No Sampling sites Lissemys punctata D/Km² Pangshura smithii D/Km² Total % 1. Thoha Bahadur 183.2 (50.4) 15.2 67.9 (56.3) 5.6 251.1 51.9 2. JabairPur 65.5 (18.0) 5.4 - - 65.5 13.5 3. Maswal Dam 55.6 (15.3) 4.6 52.3 (43.3) 4.3 107.9 22.3 4. KallarKahar Lake 59.1 (16.2) 4.9 - - 59.1 12.2 Total 363.3 30.1 120.6 9.9 483.6 Mean± S.E 90.8±30.8 7.5±2.5 60.1±5.5 0.9±1.8 Relative abundance 75.1% 24.9 Frequency (F) NT/TT 4/4=1 2/5=0.5 * NT= No. of transects in which turtles were observed, TT= Total no. of transect, F= Frequency=NT/TT, D- Density /Km². *Brackets denote % population of the species at that particular site. *183.2 is total number of turtles captured from Samli T.B Sanatorium site while 50.4 is relative percentage (%) from sixteen sites.

Table 3.8 Relative abundance of freshwater turtle species from four selected sampling sites of district Chakwal during summer/spring (total time spent at each site is 3 hours, no of attempts made by net is 10 on 1Km transect visits 24) by using CMR method.

Soft–shell turtle Hard shell-turtle

Sr. No Selected sites (Lissemys punctata) D/Km² (Pangshura smithii) D/Km² Total %

1. Thoha Bahadur 95.2 (35.6) 7.9 36.8 (44.8) 3.6 132 37.7

2. JabairPur 56.9 (21.2) 4.7 - - 56.9 16.2

3. Maswal Dam 67.5 (25.2) 5.6 45.2 (55.1) 3.7 112.7 32.2

4. KallarKahar Lake 47.8 (17.8) 3.9 - - 47.8 13.6

Total 267.4 22.1 82 7.3 349.4

Mean± S.E 20.5±10.2 5.5±0.8 20.5±2.9 1.8±0.0

Relative abundance 76.5% 23.4%

Frequency (F) NT/TT 4/4=1 4/2=0.5

* NT= No. of transects in which turtles were observed, TT= Total no. of transect, F= Frequency=NT/TT, D- Density /Km². *Brackets denote % population of the species at that particular site. *95.2 is total number of turtles captured from Samli T.B Sanatorium site while 35.6 is relative percentage (%) from sixteen sites.

Table 3.9 Population estimates of freshwater turtle species from four selected sampling sites of district Chakwal during winter/fall season (total time spent at each site is 3 hours, no of attempts made by net is 10 on 1Km transect visits 24)by using CMR method.

Soft–shell turtle Hard –shell turtle

Sr. No Selected sites Lissemys punctata D/Km² Pangshura smithii D/Km² Total %

1. Thoha Bahadur 36.8 (70.6) 3.0 4 (32) 0.3 40.8 68.4

2. JabairPur 4.6 (8.8 0.3 - - 4.6 7.7

3. Maswal Dam 1.1 (2.1) 0.0 3.5 (46.6) 0.2 4.6 7.7

4. KallarKahar Lake 9.6 (18.4) 0.8 - - 9.6 16.1

Total 52.1 4.1 7.5 0.5 59.6

Mean± S.E 13.0±8.1 1.0 ±0.6 1.8±0.1 0.1±0.0

Relative abundance 87.4% 12.5 %

Frequency (F) NT/TT 4/4= 1 2/4= 0.5 * NT= No. of transects in which turtles were observed, TT= Total no. of transect, F= Frequency=NT/TT, D- Density /Km². *Brackets denote % population of the species at that particular site.

During winter/fall season, total numbers of turtles recorded at all selectedwater bodies from 2012 to 2014 was just 59 (as compared to 349 during summer season); Relative abundance of Lissemys punctata was 87.4% and

Pangshura smithii 12.5% (Table 3.8). Maximum population of turtles (40) during this season was recorded at “Thoha Bahadur” sampling site while minimum (4.6) at

“JabairPur” site (Table 3.9; Fig. 3.6).

3.4.2.2 Population estimates of turtles using direct count method

3.4.2.2.1 Abundance of freshwater turtles

In district Chakwal, a total of four (N=4) sampling sites or water bodies were sampled for estimating populations of different turtle species (Table 3.10) using direct count method. Total numbers of turtles recorded from all sampling sites of this district by direct count method were 534. Relative abundance of L. punctata was found to be (69.6%) and that of P. smithii was 30.3 % (Table 3.10).

Highest population of turtles was recorded at “Thoha Bahadur” sampling site (36.8

%) while minimum at Kallar Kahar Lake sampling site (12.9 %).

3.4.2.2.2 Frequency (F) and density (D) of turtles

The direct count method used showed frequency (F) of direct sighted

Lissemys punctata (1) being most frequent species in the district Chakwal during the study period, and less frequent (0.5) was Pangshura smithii (Table 3.10).

Density estimates of the two turtle species from this district by direct count method revealed highest average density of 12.3±0.9/km² for Lissemys punctata and

9.0±0.1/km² Pangshura smithii (Table 3.10).

3.4.2.2.3 Seasonal variation in turtle populations by direct count method

During summer/spring season (from 2012 to 2014), a total N=474 turtles were recorded from four sites of district Chakwal using direct count method;

Lissemys punctata represented 69.4% and Pangshura smithii was 30.5%.

Maximum population (172) of turtles was recorded at the “Thoha Bahadur” site and minimum (60) at “Kallar Kahar Lake” site (Table3.11).

However, during winter/fall season, numbers of freshwater turtles recorded at selected water bodies from 2012 to 2014 was just 60 (as compared to 534 during summer season); Relative abundance of Lissemys punctata was 71.6% and

Pangshura smithii as 28.3% (Table 3.12). Maximum turtles (41.6) during this season were recorded at“Thoha Bahadur”site while minimum (15.0) at “Kallar

Kahar Lake” site (Table 3.12; Fig. 3.7).

3.4.2.2.4 Comparison of turtle populations by CMR and direct count methods

A total of N=3004 turtles were observed/ recorded using direct count method during the current study period from sixteen different sampling sites of the two districts of Pothwar Plateau. Whereas lower numbers of turtles (n= 1681) were recorded by CMR method (Table 3.13; Fig. 3.8).

The results of populations of turtles estimated using CMR and direct count method during the study period were compared usingtwo way Analysis of

Variance(ANOVA), a significant difference (p < 0.05) was found between direct count and CMR methods applied for study sites and also between four turtles species at selected sites (Table 3.14). Direct count method gives higher

Table 3.10 Population estimates of freshwater turtles from four selected sampling sites of district Chakwal during 2012-2014 by using direct count method.

Soft- shell turtle Hard- shell turtle

Sr. No Selected sites Lissemys punctata D/Km² Pangshura smithii D/Km² Total %

1. Thoha Bahadur 119 (31.9) 9.9 78 (48.1) 6.5 197 36.8

2. Jabair Pur 82(22.0) 6.8 - - 82 15.3

3. Maswal Dam 102 (27.4) 8.5 84 (51.8) 7.0 186 34.8

4. Kallar Kahar Lake 69 (18.5) 5.7 - - 69 12.9

Total 372 30.9 162 13.5 534 Mean± S.E 148.8±11.0 12.3±0.9 40.5±2.1 9.0±0.1 Relative abundance 69.6% 30.3% Frequency (F) NT/TT 4/4=1 2/5=0.5

estimates as compared to CMR method. The significant differences among populations of different turtle species were calculated by using Tukey’s Post-Hoc test (Appendix I, 1I, III and IV).

3.4.3 Morphometryof the Captured Specimens of Freshwater Turtle Species

Body measurements of the freshwater turtles captured by using cast net from the selected sampling sites of the study area were recorded for their physical parameters like body weight, carapace length and width, plastron length and width, height of shell from the ground (Fig. 3.9).

3.4.3.1 Indian flap shell turtles (Lissemys punctata)

A total 1006 specimens of Lissemys punctata were captured during the current study period; N= 375 were male, N= 424 females and N= 207 were juveniles. Females of this species were found heavier (2510± 3.3 g) on average as compared to males (2130± 4.3 g). Similarly, measurements of carapace and plastron (length and width) as well as height of the shell were found greater in females than those of males. However, Juveniles of this species showed, obviously, lesser body measurements in comparison with adults, average body weight being

880± 2.1(Table 3.15).

3.4.3.2 Indian softshell turtle (Nilssonia gangetica)

A total 491 specimens of Nilssonia gangetica were captured from the study area;

N= 180 were male, N= 207 female and N=104 were juveniles. Average body weights of females of this species were also found greater (4300± 6.5 g) than those of males (3500± 6.1 g). Similarly, measurements of carapace and plastron (length

Table 3.11 Relative abundance of freshwater turtles from four selected sampling sites of district Chakwal during summer/spring by using direct count method.

Soft- shell turtle Hard- shell turtle Sr. No Selected sites L.ssemys punctata D/Km² (Pangshura smithii) D/Km² Total % 1. Thoha Bahadur 102(31.0) 8.5 70(48.2) 5.8 172 36.2

2. JabairPur 72(21.8) 6.0 - - 72 15.1 3. Maswal Dam 95 (28.8) 7.9 75 (51.7) 6.2 170 35.8 4. KallarKahar Lake 60 (18.2) 5.0 - - 60 12.6 Total 329 27.4 145 12.0 474 Mean± S.E 131.6±9.0 10.9±0.8 96.6± 20.9 8.0±0.1 Relative abundance 69.4% 30.5% Frequency (F) NT/TT 4/4=1 2/5=0.5

Table 3.12 Relative abundance of freshwater turtle species from four selected sampling sites of district Chakwal during winter/fall (total time spent at each site is 3 hours, no of attempts made by net is 10 on 1Km transect visits 24) by using direct count method.

Soft- shell turtle Hard- shell turtle Sr. No Selected sites Lissemys punctata D/Km² Pangshura smithii D/Km² Total % 1. Thoha Bahadur 17 (39.5) 1.4 8 (47.0) 0.6 25 41.6

2. JabairPur 10(23.2) 0.8 - - 10 16.6 3. Maswal Dam 7 (16.2) 0.5 9 (52.9) 0.7 16 26.6 4. KallarKahar Lake 9(20.9) 0.7 - - 9 15.0 Total 43 3.4 17 1.3 60 Mean± S.E 17.2 ± 2.1 1.4±0.1 11.3± 0.3 0.5±0.1 Relative abundance 71.6% 28.3% Frequency (F) NT/TT 4/4=1 2/5=0.5

Table 3.13 Comparison of populations of freshwater turtle species by CMR and direct count methods during 2012 to 2014 at sixteen (16) selected sampling sites of the Pothwar Plateau.

CMR Direct count

Sr. No L. punctata N. gangetica P. smithii P. tecta L. punctata N. gangetica P. smithii P. tecta

1. Total number of 1043.3 439.4 547.2 135.2 1256 575 915 258 individuals

2. Mean± S.E 69.5 ± 8.6 43.9 ± 4.9 47.9 ± 3.5 27.0 ± 1.6 63.7 ± 4.7 57.5 ± 4.6 83.1 ± 8.2 51.6 ± 3.3

3. Relative 48.1% 20.2% 25.2% 6.2% 41.8% 19.1% 30.4% 8.5% abundance

4. Frequency (F) 15/16= 0.9 10/16=0.6 11/16=0.6 5/16= 0.3 15/16=0.9 10/16=0.6 11/16=0.66 5/16=0.3

5 Average Density 86.9/Km² 36.6/Km² 45.5/Km² 11.2/Km² 104.6/Km² 47.9/Km² 76.2/Km² 21.5/Km² /Km²

Table 3.14 Statistical comparison of turtles populations by using two methods (CMR and direct count) at sixteen selected sampling sites of the Pothwar Plateau by Two- way Analysis of Variance (ANOVA)

Sr. Species Source of df SS MS F p-values

No variation

1. L.punctata Location 15 20223.9 1348.2 25.7 5.290e-08 ***

Method 1 1953.1 1953.1 37.2 2.011e-05 ***

2. N. gangetica Location 15 27626 1841.7 26.6 4.178e-08 ***

Method 1 288 288.0 4.1 0.05936

3. P. smithii Location 15 39059 2603.9 6.0 0.0006086 ***

Method 1 3894 3894.0 9.0 0.0088273 **

4. P. tecta Location 15 12249.9 816.6 13.5 4.166e-06

Method 1 378.1 378.1 6.2 0.0242 *

Residuals 15 902.9 60.1

Figure 3.8 Comparative populations of freshwater turtles (all species) by CMR and Direct count methods from sixteen sampling sites of Pothwar Plateau during 2012 to 2014. Values represent mean ± SE.

A B

C D

Figure 3.9 Photograph showing body measurments of Lissemys punctata; A)

Carapace length, B) Carapace width, C) Plastron width and D) Shell height of

Pangshura tecta.

Table 3.15 Body measurements of four turtle species captured from all selected sampling sites of Pothwar Plateau (N=2216)

Species Sex Body weight (g). Carapace Plastron Shell height (cm.) Length width Length width (cm.) (cm.) (cm.) (cm.) Lissemys punctata Males (N= 375) 2130± 4.3 14.3±0.5 12.2±0.2 11.9 ±0.3 8.9± 0.4 2.1 ± 0.1 Females(N=424) 2510± 3.3 16.7± 0.3 14.2±0.5 12.5± 0.1 10.9± 0.4 2.4 ± 0.1 Juveniles (N= 207) 880± 2.1 8.1±0.8 7.4±0.4 6.8± 0.3 5.9±0.6 0.8 ± 0.5 Nilssonia gangetica Males (N= 180) 3500± 6.1 17.1± 1.1 15.8±0.9 16.9± 1.1 14.7±0.4 2.5±0.5 Females(N=207) 4300± 6.5 18.7± 1.2 16.6± 0.7 17.4± 1.3 15.9± 1.4 2.7±0.5 Juveniles (N= 104) 1360± 5.7 12.5± 0.5 10.4± 0.2 11.6± 0.9 9.9± 0.3 1.1±0.1 Pangshura smithii Males (N= 215) 2830± 3.3 15.6± 0.8 13.3± 0.2 14.9± 0.4 11.7± 0.5 2.2± 0.5 Females(N=231) 3100± 3.1 16.9± 0.5 14.6± 0.5 15.4± 0.3 14.9± 0.3 2.3± 0.4 Juveniles (N= 115) 890± 0.8 9.5± 0.3 7.4± 0.5 8.6± 0.4 6.9± 0.1 1.3±0.1 Pangshura tecta Males (N= 67) 2170± 4.6 15.9± 0.9 14.3±0.3 14.8± 0.8 10.9± 0.3 2.4± 0.9 Females(N=65) 2590± 4.8 16.3± 0.1 15.2± 1.1 15.4± 1.3 13.9± 0.5 2.6± 1.1 Juveniles (N= 26) 690± 0.9 8.7± 0.5 7.1± 0.3 7.7± 0.3 6.5± 0.3 1.9± 0.7

and width) and the shell height were greater in females, while Juveniles showed much lesser body measurements in comparison with adults, and their average bodyweight being 1360± 5.7 g (Table 3.15).

3.4.3.3 Brown river turtles (Pangshura smithii)

A total 561 specimens of Pangshura smithii were captured during the current study period; out of these N=215 were males, N=231were females and

N=115 juveniles. This species also showed heavier females (3100± 3.1g), on average, as compared to male (2830± 3.3g). Similarly, measurements of carapace and plastron (length and width) were found greater in females than those of males.

Whereas, average body weight of juveniles was found to be 890± 0.8 g (Table

3.15).

3.4.3.4 Indian roofed turtles (Pangshura tecta)

The numbers of specimens of Pangshura tecta during the current study were N= 158 including N= 67 were found males, N= 65 females and N= 26 juveniles. Female of this species showed greater body weights (2590± 4.8 g) compared to males (2170± 4.6 g). Similarly, carapace and plastron (length and width) measurements were greater in females whereas average body weight of juveniles was 690± 0.9 g (Table 3.15).

3.4.4 Statistical Analysis of Body Parameters of the Turtles

The body measurements of four freshwater turtle species were compared using Two-way Analysis of Variance (ANOVA), a significant difference

(p < 0.05) was found between body weights (kg); carapace length, width; plastron

Table 3.16 Statistical comparison (gender-wise) of body measurements of four different freshwater turtle specie by using Two- way Analysis of Variance (ANOVA) that were collected from Pothwar Plateau.

Source of Variance df SS MS F p-Value

Body weight 2 1.043E7 5217308.333 13.245 0.002 Carapace length 2 125.112 62.556 29.951 0.000

Carapace width 2 114.065 57.032 29.168 0.000

Plastron length 2 103.940 51.970 12.327 0.003

Plastron width 2 89.527 44.763 9.848 0.005

Shell height 2 3.455 1.728 18.345 0.001

Total 11 141.882

Table 3.17 Statistical comparison of body measurements of four different freshwater turtle species collected from Pothowar Plateau by using Two- way

Analysis of Variance (ANOVA).

Source of Variance df SS MS F p-Value

Body weight 3 300509.1 1001697.2 0.730 0.562

Carapace length 3 15.9 5.33 0.333 0.802

Carapace width 3 15.6 5.2 0.359 0.784

Plastron length 3 36.1 12.06 0.913 0.477

Plastron width 3 37.4 12.4 1.07 0.413

Shell height 3 0.469 0.159 0.326 0.807

Total 12 2115.65

length, width and shell height of freshwater turtle species and between the genders; and non-significant difference (p ˃ 0.05) was found between four turtle species

(Table 3.16, 3.17).

3.4.5 Sex Ratio

During current study, a total of 1006 specimens of Lissemys punctata were captured from sixteen selected sites of Pothwar Plateau. Out of these (375) individuals were found to be males while 424 females and remaining were juveniles (Table 3.15). It gave a sex ratio of Indian flap shell turtle as 1.13:1 (M: F, respectively).

A total 491 of Nilssonia gangetica were captured by cast net (Table 3.15).

One hundred and four of the captured individuals of this species were juveniles.

The remaining individuals were found to be in the ratio of 1.15:1 (M: F, respectively). Pangshura smithii captured in the cast net were (561); two hundred and fifteen were male while two hundred and thirtyone were female, giving sex ratio 1.07: 1 (M: F, respectively). Pangshura tecta captured specimens (158) out of this 67 were found males, 65 females sex ratio was 1.03:1 (M: F, respectively)

(Table 3.15).

3.5 DISCUSSION

Freshwater turtles have served as an important food resource in Asian tropical and sub-tropical parts of many countries. However, several countries of the region have made only few efforts for protection and management of these freshwater turtle species. In Pakistan, very few studies have focused turtles for their

population estimates, although many checklists of the turtle species are available from different parts of the country. Current study estimated population abundance of different species in the Pothwar Plateau. Two methods (CMR and direct count) were employed for estimating turtle populations in the study area.

Results showed a total of 3004 and 1681 turtles were captured by CMR, and direct count method, respectively, belonging to two families viz., Trionychidae

(Lissemys punctata and Nilssonia gangetica) and Geoemydidae (Pangshura smithii and Pangshura tecta) were recorded from all sixteen selected sampling sites. Four different species were recorded in water bodies of the district Rawalpindi whereas only two species were found in district Chakwal. Both methods used revealed

Lissemys punctata as most abundant turtle species, followed by Pangshura smithii, and Nilssonia gangetica while Pangshura tecta was found least abundant.

Some earlier published studies are available about the turtle species occurring in the country. For example, Khan (2015) observed six freshwater turtle species in district Thatta of Sindh province; among those Lissemys punctata and

Pangshura smithii, were found abundant. Chitra indica, Nilssonia gangetica and

Nilssonia hurum were common, Hardella thurjii was less common.Similarly, Khan et al. (2015) reported four species of freshwater turtles from district Badin (Sindh);

Lissemys punctata (37%,) and Chitra indica (40%), were abundant, Nilssonia hurum (18%) was common while Geoclemys hamiltonii (4.31%), was found rare.

Similarly, another study by (Safi and Khan, 2014) reported five freshwater turtle species in district Pehawar of the Khyber Pakhtoonkhwa (KP) province; L. punctata as (34.05%), N. gangetica (22.09 %), A. hurum (18.40%), P. smithii

(15.64%), and C. indica (9.82%). Akber et al. (2006) studied the on distribution of fresh water turtles in Punjab, recorded a total of 3528 specimens belonging to eight different species Pangshura smithii (43.62%) and P. tectum (42.06%) were found abundant, whereas; Hardella thurjii (0.88%) and Chitra indica (0.54%) were found rare. An other study done by Safi et al. (2014) reported a total of 365 Individual turtles from district Charsadda of Khyber Pakhtunkhwa (KP) province and the most abundant turtle species found was Lissemys punctata (30.95%), followed by

Nilssonia gangetica and Nilssonia hurum (27.39% and 18.90%, respectively,

Hardella thurjii (1.36%) and Pangshura tecta (3.01%) were rare,whereas the status of Pangshura smithii (10.13%) and Chitra indica (8.21%) were frequent, Lissemys punctata (30.95%) was found common.

Average population density of the four recorded freshwater turtle species was estimated in the current study using data from sixteen sampling sites of the study area by applying two methods (CMR and direct count) Results showed an average densityof Lissemys punctata 86.9/Km², Nilssonia gangetica 36.6/Km²,

Pangshura smithii 45.5/ Km² and Pangshura tecta 11.2/ Km² by direct count method. However, the CMR method used estimated average density of Lissemys punctata as 104.6/ Km², Nilssonia gangetica 47.9/ Km², Pangshura smithii 76.2/

Km² and Pangshura tecta 21.5/ Km². These results indicate that maximum population density in the study area occurs for Lissemys punctata and minimum for

Pangshura tecta. An earlier study by Mahmood et al. (2012) had reported average density of freshwater turtles in Korang River Islamabad- Rawalpindi, as being

38.3individuals/km²; highest population density was recorded for Indus mud turtle

(Lissemys punctata). Seasonal populations of the freshwater turtles were found

different (summer and winter populations); the summer population of turtles was recorded high while winter population was found low in the study area. (Souza and

Abe, 2001) studied that difference in populations of turtles between two seasons is most probably due to the well known phenomenon of hibernation that occurs in turtles (being reptiles and ectothermic. Since freshwater turtles cannot maintain body temperature constant to a certain level, therefore, during winter season when environmental temperature falls too low, these species undergo the hibernation.

This fact was also reported by Gibbons et al. (1990) on seasonal variation in populations of Painted turtles; hibernated overwinter and active during summer.

Jackson (1970) had reported that temperature is known to control both feeding and activity of most heterothermic species. In general, aquatic turtles are not active during winter months (Bury, 1979), with exceptions in populations inhabiting tropicaland subtropical areas. Occurrence of winter basking activity are restricted to warm spells or calm, sunny days (Moll and Legler, 1971).

The direct count method used for estimating turtles populations recorded greater numbers of turtles (N=3006) while CMR method used recorded lower numbers of turtles (N= 2195) There are so many suppositions about the authenticity of two methods. In the current study, CMR method showed low population estimation of turtle species, but most importantly this could be due to low captured success. Similarly there is also a question mark why population estimations of turtle species were high by direct count method; this could be due to the fact that turtles specimens were double counted due to human eye error Vaughn et al. (1997). Smith et al. (2000) had shown that visual observation method allows a good rough estimation of animal populations but suggested both methods as

reliable, suitable and easily applicable for wildlife population estimations.

Body measurements of four different turtle’s species recorded from sixteen sampling sites showed clear sexual dimorphism between males and females of all turtle species studied. Average body weight, carapace (length and width) and plastron (length and width) size were found greater in females in comparison with males. Regarding overall body size, Nilssonia gangetica species was found heaviest (4300 g) and largest among the four turtle species recorded during the current study, followed by Pangshura smithii (3100 g) while the other two species showed moderate body weight. Earlier on Kayani et al. (2015) recorded three freshwater turtle species from Rawalpindi region, Lissemys punctata, Nilssonia gangetica and Pangshura smithii. Their average body weights were 974.8 g, 2276 g and 510 g, respectively, for Lissemys punctata, Nilssonia gangetica and

Pangshura smithii. They also reported Nilssonia gangetica as the largest and heaviest turtle species among the three, very similar to results of the current study.

Similarly, carapace and plastron length and width were greater for Nilssonia gangetica. However, in the current study, females of this species were found heavier (3510 g) than earlier on reported. Similarly, (Das, 1995) recorded maximum carapace length of P. tecta as 23.0 cm where the sex and weight were not mentioned. Minton (1966) indicated that the male was 17.0 cm and its shell was 10.5 cm in Pangshura tecta. Whereas Moll (1987) mentioned that average carapace length of male was 6.6 cm, carapace width 5.4 cm, plastron length 6.1 cm, plastron width 4.1cm, body weight 540g found in P. tecta. Frazier (1997) stated that average carapace length of P. tecta was 10.7 cm, shell was 4.6 cm and body weight was 110g without mentioning the sex.

3.6 CHAPTER SUMMARY

Freshwater turtles are the most dynamic group of wetlands associated fauna

They play same role in the aquatic habitat which is played by vultures in the terrestrial areas. Eight species of freshwater turtles belonging to two families and six genera are reported from Pakistan. The two families of turtles in Pakistan include Geoemydidae which consists of hard-shelled turtles while the second family Trionychidae comprises of softshell turtles. Soft shelled turtles of Pakistan are illegally exported to China and other countries for their use in Traditional

Chinese Medicines. The current study estimated populations of four turtle species recorded form the Pothwar Plateau. Two different methods were employed viz

“Capture, Mark and recapture (CMR)” and “direct count” methods.

A total of 2166 specimens of freshwater turtles were recorded using CMR method; Lissemys punctata was found most abundant species (48.1%), and most frequent turtle species in the study area, followed by Pangshura smithii

(25.2%),and Nilssonia gangetica (20.2%) while the least common was Pangshura tecta (6.2%).A total of 3004 specimens of turtles were counted during the study period using direct count method; again Lissemys punctata was the most abundant species (41.8%), followed by Pangshura smithii (30.4%), and Nilssonia gangetica

(19.1%) whereas Pangshura tecta (8.5%) was found least common.

Estimates of average population density (per square kilometer) of freshwater turtles were (20.8/ Km²) by direct count method as compared to CMR

(15.0/Km²) method during study period. Seasonal populations (summer and winter) of turtles species were found significantly different; In summer/spring season,

turtles populations were high. However, during winter/fall season, turtles populations were found low.

Body measurements captured specimens of freshwater turtle species from selected sites were recorded. Results showed Nilssonia gangetica species was found heaviest (4300 g) in body weight and largest among the four turtle species recorded during the current study.

Chapter 4

INVESTIGATION OF THE FOOD HABITS OF FRESHWATER TURTLE

SPECIES OCCURRING IN POTHWAR PLATEAU

4.1 INTRODUCTION

Turtles play vital role in the ecosystem by feeding on a variety ofaquatic and semi-aquatic animals, viz. worms, insects, snails, slow movingaquatic crabs, dead animals and fragments of dead bodies, and are helpful to reducing thewater pollution (Hossain and Sarker, 1995). Turtle species are also important incontrolling larval and adult stages of aquatic insects, aquatic vertebrates, trash fish, mollusks, other debris and valuable organisms (Fugler, 1984). They also perform a valuable service by acting as scavengers in water bodieskeeping the aquatic systems free from pollution (Rao, 1985).

Diets of freshwater turtles have often indicated theavailability of food resources in the territory. The diet of bottom-feeding turtles is commonly composed of the benthic macro-invertebrate fauna, insects and mollusks (Ernst and

Lovich, 2009). Freshwater turtles how feeding behavior viz. omnivore, herbivore and carnivore. Herbivorous species of freshwater turtles control the growth of aquatic weeds and other floating vegetation, and conserve healthy aquatic environment for wildlife and fish (Philip et al., 1998). Turtles also show strategic carnivore behavior by obtaining food from anextensive variety of sources including planktons, nektons, benthic macro-organisms, carrion, and terrestrial organisms that fall upon the water (Richard and Wensing, 1986). The plant matter forms a notable proportionof the diet of freshwater turtle species (Carr, 1952; Pritchard,

1979). However, many turtles display some sort of selective feed, on the basis of particular resources (Mahmoud, 1968; Ernst and Lovich, 2009). Freshwater turtles display particular tactics and strategies in their feeding habits: differences in diet among sympatric species (Lagler, 1943; Brecken-ridge, 1944; Berry, 1975; Vogt,

1981).

Temperature is an important factor that controls the rate of both chemical and biochemical reactions, which in turn regulate both growth and reproduction

(Vembergand Vemberg, 1972; Prosser, 1973). Temperature fluctuation regulates both activities ecto-thermis and feeding of species. In general, aquatic turtles are not active during winter months (Bury, 1979), with exceptions in populations inhabiting tropical (Moll and Legler, 1971) and subtropical (Jackson, 1970) areas.

Examples of winter basking activity are confined to warm spells or calm, sunny days, approximately below 14 ºC temperature feeding activity usually ceases

(Ernst, 1972, 1976; Bury, 1 979).

4.2 REVIEW OF LITERATURE

Fresh water turtles play a vital role in the health of aquatic ecosystem; they act as scavengers, herbivores, and carnivores, and also by playing important role in dispersal of plants seeds (Brooks et al., 1988; Lovich, 1994). Bottom feeding freshwater turtles often indicate the structure of the benthic macro-invertebrate communities where they feed, which impact by the presence of certain bivalve mollusks (Newell, 2004).

Turtles are also important component of the ecosystem since they reduce the water pollution by feeding on dead animals and fragments of dead bodies,

along with a variety of aquatic and semi-aquatic animals, viz. worms, insects, snails, slow moving aquatic crabs, (Hossain and Sarker, 1995). Turtle species also performa valuable service to control larval and adult stages of aquatic insects, aquatic vertebrates, trash fish, mollusks, other detritus and beneficial organisms

(Fugler, 1984). In the present decade, commercial values have increased significantly that cause exploitation of natural resources (Khan, 1982).

Some researchers (Hossain and Sarker, 1993; Rashid and Swingland, 1997) used stomach contents analysis method in different countries. They were captured turtles mainly at day time from open water area in the field and were anesthetized instantly. The stomachs of the turtles were dissected out in the field and then preserved in an air tight container. A solution of 5% formalin and 1% glycerin was used as preservatives. The qualitative analysis of food items was made. The stomach contents of each turtle were separated and washed in a Petridish and categorized into two groups: (a) animals and (b) unidentified groups, by examining under the microscope. The food items were identified at species level. They reported that the turtles are totally carnivore. Das (1995), however, is of the view point that they feed on adult frogs, tadpoles, fish, crustaceans, snails, earthworm, carrions, insects and aquatic weeds. Similarly, Deraniyagala (1953) detected that turtle coming to shore at night fed on carrion or decaying matters and other animal foods.

Turtles are omnivore in their feeding behavior (Whitaker and Romulus, 1997).

Vijaya (1981) reported their carnivorous as well as herbivorous behavior, by preying upon living organisms, and scavenging on organic debris from the pond

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bottom and household garbage; often feedingon earthworms, frogs, tadpoles, wasp and beetles.

Freshwater turtles feed on aquatic and terrestrial vertebrates and terrestrial invertebrates, carrion, and a great variety of plants, seeds and algae (Souza and

Abe, 1995, 1997, 1998; Lima, Magnusson and Costa, 1997; Portal et al., 2002;

Balensifer, 2003; Malsavio et al., 2003; Souza, 2004). Bury (1986) reported the presence of prey items in stomach contents of turtles (Georges et al., 1986) belonging to the prey species from class reptilia Clemmys marmorata and

Chelodina longicollis, also differentiating the stomach contents with the prey items available in the environment. They also investigated that terrestrial preys were consumed more, probably because they fell on the water.

Strecker (1927) studied the food habits inTexas Cooter (Pseudemys texana) andreported only mollusks species (Sphaerium planorbis and Lymnaea), insects, crayfish, and other invertebrates recovered from in the digestive tracts of young P. texana. Similarly, Vermersch (1992) indicated that other members of this genus consumed more invertebrates when young but becoming less carnivorous with maturation. Some other researchers (Ernst and Barbour, 1989; Vermersch, 1992;

Ernst et al., 1994) assumed that P. texana was herbivorous when mature, although quantitative analysis of the diet was lacking.

Earlier studies (Gibbons et al., 1983; Ernst et al., 1994; Rowe et al., 2009) on common Musk Turtle (Sternotherus odoratus) have indicated that this species is widely distributed and habitat of this species was relatively shallow waters with plentiful swamp vegetation and woody detritus; it rarely used to leave the water

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even under drought conditions. Feeding habits of Sternotherus odoratus showed it as a bottom-feeding omnivore, feeding on crayfish, insects, mollusks, fish, amphipods, arachnids, algae, seeds, and other plant material throughout its range

(Berry, 1975; Marion et al., 1991; Ernst et al., 1994; Ford and Moll, 2004).

Dietary habits of many turtle species show some sort of selectivity, by often selecting for particular resources (Mahmoud, 1968; Ernst and Lovich 2009). The availability of certain food resources can also be highly affected by aquatic invasive species and can even lead to shifts in diet (Preston et al., 2012; Rush et al.,

2012). Some earlier studies (Legler, 1966; Moll and Legler, 1971) have demonstrated the feeding behavior of geotropically freshwater turtles, white-lipped mud turtle (Kinosternon leucostomum) being omnivorous in feeding behavior.

Some other turtle species like Yellow-bellied Slider Turtle (Trachemys scripta) in

Panama (Moll and Legler, 1971) and Giant Musk Turtle (Staurotypus salvinii) in

Chiapas, Mexico (Dean, 1980) have also been shown to be omnivorous. Minton

(1966) in Pakistan observed a turtle eating aquatic vegetation. But others like

Barbour and Ernst (1989) statethat turtles are omnivore.

Indian flap shell turtle (Lissemys punctata) is carnivorous in nature was studied by a number of researchers (Vijaya, 1981; Auffenberg, 1981; Whitaker and

Romulus, 1997; Deraniyagala, 1953). Similarly, Hossain et al. (2012) indicated that this species iscarnivore in nature and feeds on natural available foods, insects by digging the soil with the help of snout, neck and forelimbs. Mostly turtles moveto lands to feed on garden snails, broken the shells with the help of bony shields like teeth and consumed the flesh.

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Indian soft-shell turtles Nilssonia gangetica were collected and stomach contents flushing technique (Moll, 2004) was used to study their food habits.

Samples of stomach contents were stored in containers with 70% alcohol (Moll,

2004). Results showed that Nilssonia gangetica are known to be omnivores in their food habits Tikader and Sharma (1985) found that N. gangetia were omnivorous in nature, mainly feeding on aquatic vertebrates, but Das (1985) reported that sometimes they used to eat spoiled eggs of own species and carrion.

Cuvier (1825) also reported N. gangetica as omnivorous, ingesting mostly water plants, invertebrates and vertebrates.

The Indian roofed turtle (Pangshura tecta) has beenfound to feed all the daytime by continuous movement in the aquatic habitats’, however, sometimes they depended on roots of floating vegetation and also on semi submerged weeds and fed stems or tender leaves by extending their neck. By rapid crawling at the bottom of water, the turtle, sometimes, tried to grab carrion from the down water body (Gray, 1831). This turtle species isan important biotic component of freshwater bodies and mostly feeds on meat (Das, 1995 and Daniela, 2000).

According to some researchers (Smith, 1931; Minton, 1966; Das, 1985) brown roofed turtle (Pangshura smithii) are omnivorous but prefer flesh; eat meat of fish, frogs and fruits of several kinds. Tikader and Sharma (1985) demonstrated eating behavior of P. smithii, showingboth types being herbivorous as well as carnivorous; most of the time feeding on vegetation, but sometime also consuming animals

4.3 MATERIALS AND METHODS

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Investigation of food habits of the freshwater turtles inhabiting the study area were carried out by stomach contents flushing method.

4.3.1 Stomach Contents Flushing

Freshwater turtles belonging to four different species were live captured using cast net at selected sampling sites at day time. After the capture, A “Naso– gastric” tube attached with a plastic syringe was introduced through the esophagus into the stomach of the turtle. The neck muscles of the turtles were partially anesthetized by injecting anesthesia (30 mg/kg body weight Zoletil 50; Vivric). In order to induce the regurgitation of food items, approximately 100-200 ml tap water was delivered into the stomach. The pyloric end of the stomach can normally be felt. The entire content of the syringe was flushed into the stomach; the empty syringe was separated from the tube, refilled with water and connected to the tube again to repeat the flushing procedure. This procedure was repeated 5 times as long as stomach contents are forced out. When no more stomach contents were left, the animal was flushed for one last time. The water with stomach contents was then drained into a sieve. The food items were picked up with forceps and fixed in 70% ethanol in a vial. Then the stomach contents were directly washed and segregated with tap water from the gauze. Each sample was segregated into different categories including animal matter (insects and other invertebrates), plant matter like vegetation (including algae), and other materials. Each item was identified to the lowest possible order level (Legler, 1977) (Fig. 4.1 A-F).

4.3.2 Collection of Reference Plant Samples

A reference collection of plants was carried out from the same study area simultaneously along with the stomach contents flushing samples. The collected

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plant samples were identified and used for preparation of reference slides for comparison.

4.3.3 Micro-Histological Analysis of Stomach Content Flushing Plants

Analysis of stomach contents samples by micro-histological techniques is the most commonly used method for determining the botanical composition of herbivore diet (Alipayo et al., 1992). The following steps were followed for the micro histological analysis of plant samples obtained from stomach contents flushing of freshwater turtle species The plant materials were separate from stomach content samples then washed in following water and soaked in a soaking solution (1 part distilled water, 1 part Ethyl alcohol, 1 part glycerin) for overnight.The samples were ground in Vertis Homogenizer (50%) of the samples were transferred to a labeled test tube and (5%) warm sodium hydroxide solution was added to it. After that test tube was heated in a boiling water bath for 4 to 6 minutes. The particles were allowed to settle before removing the supernatant dark fluid and this treatment was, repeated until a relatively clear solution produced then the material was washed with distilled water for 2 minutes. It was dehydrated through a series of 25%, 50 %75 % and 100 % alcohol treatments, each for 10 minutes. The alcohol was removed through a series of xylene and alcohol mixtures

(25%, 50 %, 75 % and 100 % for overnight). Afterkeeping the material into xylene

(100 %) for overnight, it was transferred to a clean glass slide and was evenly spread and mounted indibutyl phthalate xylene (DPX) mounting medium under a cover slip. The same procedure was followed for preparation of slides of reference

Plants collection, except for using 10 percent NaOH solution in step five. Five

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A B

C D

E F

Figure 4.1 Photographs taken during the stomach contents flushing procedure, (A) showing stomach contents flushing apparatus (B) Anesthesia injection (C) Injecting the Anthesia into the turtles muscles, (D) Passing Naso-gastic tube into the turtle’s stomach, (E) Filling the turtle’s stomach with water, (F) showing stomach contents being forced out.

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slides were prepared for each sample ten microscopic fields per slide were observed. The photographs taken of cells from prepared reference were used to match the cells in the sample slides. The “frequency of occurrence” of plant species was calculated as the numbers of specific plant species in the sample slide/number of total plant species identified in sample slide. The relative frequency of each food item was calculated as total number of fragments identified for a given food item, and were divided by the total number of all counts made in the sample slide, multiplied by 100. The results were expressed as total number of dietary species in the sample.

4.3.4 Seasonal Variation in Diet Composition of Each Turtle Species

For investigating seasonal variation in the diet composition of each freshwater turtle species, the samples were pooled season-wise including summer, winter and rainy seasons. In addition, prey species richness (S), Diversity Index

(H') of prey species and Evenness Index (E), were calculated from the data on seasonal variation in the diet composition of each turtle species as follows:

4.3.4.1 Prey species richness index (S)

It was calculated by taking into account the total numbers of plant and animal prey species consumed by each turtle species in a specific season.

S= number of prey species present per sample

4.3.4.2 Diversity index (H')

Diversity index (H') of prey species was calculated by Shannon’s index:

H' = [Pi× logPi]

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Where Pi is the proportion (n/N) of individuals of particular prey species (n) divided by total number of prey species (N).

4.3.4.3 Evenness index (E)

Evenness index (E) refers to how close in numbers each species in an environment is. The Shannon diversity index (H) is another index that is commonly used to characterize species diversity in a community. Evenness index was

Calculated using Pielou’s formula:

E= H'/ Log of S

Where 'H' representing Shannon’s index and S the prey species richness index.

4.3.5 Statistical Analysis

Prey items and different species of freshwater turtles were compared by using twoways Analysis of Variance (ANOVA). The significant results of ANOVA were later on processed through Box plot, thesignificance was alsocalculated by using Post-Hoc test (LSD) and Duncan test. Insect orders, plants and turtle species were compared through ANOVA, using General Linear Model (GLM SPSS-17)

Analysis. The p value less than 0.05 was considered as significant.

4.4 RESULTS

For food habits investigations of freshwater turtle species, stomach flushing method was used; the specimens were collected from selected sampling sites and

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stomach contents were collected on spot by applying the procedure and preserved in 70% ethanol for later analysis. The size of stomach contents collected varied from 10 g to 55 g at maximum.

4.4.1 Collection of Stomach Samples

A total of N = 410 samples of stomach contents of four freshwater turtle species were collected from various water bodies of the study area; including N=20 samples each from sites from site I, XIII and XV; N=40 samples eachfrom sites II,

III, V, VI and IX; N=10 samples each from sites IV, XIV and XVI; and N=30each from sites VII, VIII, XI and XII.

The collected samples included N=150 samples of L. punctata (having average body weight 3.1±1.1 kg), N= 100 samples of N. gangetica (average body weight 4.8 ±0.1 kg), N=110 samples of Pangshura smithii (average body 2.9± 0.1 kg) and N= 50 samples of Pangshura tecta (average body weight was 2.8±0.3 kg).

4.4.2 Stomach Content Analysis

Analysis of the stomach contents samples (N=410) in the laboratory revealed five different kinds of prey items consumed by freshwater turtles of the study area including insects earthworm, snails, plant parts, algae, in addition to soil particles and unidentified material (Fig. 4.2).

4.4.3 Percent Frequency (% F) and Percent Volume (% V) Occurrence of

Prey Items

A total of five food items were identified in the stomach contents of the

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A B

C D

E F

Figure 4.2 Prey items recovered from stomach contents of freshwater turtle species (A) insects body parts (B) earthworms, (C) snails, (D) plant material, (E) algae, (F) soil particles, (G) unidentified material.

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Table 4.1 Percent frequency (% F) of the prey items recovered from the stomach contents of freshwater turtles collected from selected study sites.

Prey item L.Punctata N. gangetica P. smithii P. tecta Mean % (n=150) (n=100) (n=110) (n=50) (n=410)

Insects parts 93.3(140) 85 (85) 71.8 (79) 80 (40) 83.9 (344) Earthworm 87.3 (131) 72 (72) 60.9 (67) 66 (33) 73.9 (303) Snails 65.3 (98) 66 (66) 45.4 (50) 50 (25) 58.2 (239) Plant parts 40 .6(61) 20 (20) 59.0 (65) 46 (23) 41.2 (169) Algae(Oscillatoria 20 (30) 20 (20) 31.8 (35) 40 (20) 25.6 (105) acuminate) Soil particles 32.6 (49) 33 (33) 48.1 (53) 46 (23) 38.5 (158) Unidentified 25.3 (38) 15 (15) 17.2 (19) 20 (10) 20 (82) Material

Table 4.2 Percent volume (% V) occurrence of the prey items recovered from the stomach contents of freshwater turtles collected from the study area.

Prey items L. punctata N. P. P. Mean % (n=150) gangetica smithii tecta (N=410) (n=100) (n=110) (n=50) Insects parts (%) 36.2 30.4 22.4 16.6 26.3

Earthworm (%) 30.8 25.8 12.6 9.8 19.9

Snails (%) 20.9 19.7 12.7 11.6 16.1

Plant parts (%) 9.1 8.6 12.5 10.7 10.2 algae 10.2 10.1 15.7 9.8 11.4 (Oscillatoria acuminate) (%) Soil particles (%) 8.2 7.9 7.4 5.6 7.5 Unidentified material (%) 9.9 8.8 8.5 6.7 8.4

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Figure 4.3 Mean percent volume (%V) occurrence of prey items recovered from the stomach contents of four species of freshwater turtles captured fromthe study area (A) L. punctata (B) N. gangetica (C) P.smithii (D) P.tecta.

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Figure 4.4 Two-ways Analysis of Variance (ANOVA) Box plot comparison showed that prey items consumption was significantly different among four freshwater turtle species.

Figure 4.5 Two-way Analysis of Variance (ANOVA) showing Box plot giving comparison that different prey items significantly differ from each other.

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Figure 4.6 Two-way Analysis of Variance (ANOVA) showing Box plot giving comparison that different freshwater turtle species differ in their food habits significantly among themselves.

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freshwater turtle species captured from the Pothwar Plateau; percent frequency (%

F) of prey items recovered included insect body parts 83.9 % (344), earthworms

73.9 % (303), snails 58.2 % (239), plant parts 41.2 % (169), and algae 25.6 %(105), in addition soil particles 38.5 % (158) and unidentified material 20 % (82) (Table

4.1).

The percent volume (% V) occurrence of food items in stomach contents of all four freshwater turtles studied showed (Table 4.2) insects body parts in highest quantity, followed by earthworms and snails, while plants parts and algae were also represented in reasonable volume. On average, insects contributed 26.3% (by volume), followed by earthworm (19.9%), snails (16.1%), algae (11.4%), and plants (10.2%) (Table 4.2; Fig. 4.3 A, B, C and D).

Statistical analysis (Two- way Analysis of Variance, ANOVA) showed significant (p< 0.05) difference between consumption of prey items consumed by four turtle species (Fig. 4.4). Similarly, consumption of prey items was significantly (p< 0.05) different to each other (Fig. 4.5). The significance difference also found within freshwater turtle species wascalculated by using Box plot comparison (Fig. 4.6; Appendix V, VI).

4.4.4 Seasonal Variation in Food Consumption of Freshwater Turtle Species

Food habits of four freshwater turtle species were variable during three different seasons of years including winter, summer and rainy seasons (Table 4.3).

4.4.4.1 Indian flapshell turtle (Lissemys punctata)

Analysis of N=50 samples each for each of the three seasons (winter,

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Table 4.3 Seasonal variation in consumption of different food items (g) by four turtle species in the Pothwar Plateau.

Species Seasons Insects Earthworms Snails Plant Algae Soil particles Unidentified

material material

Lissemys punctata Winter 25.5 ± 3.3 22.8± 1.4 19.7±1.5 11.1±1.2 7.8± 1.4 6.5±1.4 6.6±1.0

Summer 29±2.1 23±2.8 19.8± 1.3 9.9± 2.0 9.8±2.0 5±0.8 3.5±0.4

Rainy 30.2±2.0 24±1.9 17.3±1.3 9.2±0.5 7.8±0.3 5.2±0.3 6.3±0.5

Nilssonia gangetica Winter 22.4±1.5 20.7±0.6 18.7±0.5 12.7±0.7 8.9±0.9 9.9±0.7 6.7±0.3

Summer 24.3±1.1 20.9 17.6±1.7 11.7±0.3 8.9±0.4 9.9±0.2 6.7±0.4

Rainy 31.2±0.8 21.4±1.2 15.4±1.6 13.4±0.5 9.8±1.1 3.9±0.9 4.9±1.1

Pangshura smithii Winter 19.8±0.1 29.5±0.3 17.6 ±3.8 12.6±0.4 7.8±0.6 6.4±0.4 6.3±0.4

Summer 20.9±0.5 32.9±0.5 12.5±0.6 12.6±1.4 8.8±1.4 5.0±1.2 7.7±0.9

Rainy 12.6±1.6 12.3±1.6 26.8±3.7 27.2±2.2 8.8±0.8 8.8±0.8 4.0±0.3

Pangshura tecta Winter 19.8±0.1 12.6±0.4 17.6 ±3.8 29.5±0.3 7.8±0.6 6.3±0.4 6.4±0.4

Summer 12.6±1.4 32.9±0.5 12.5±0.6 20.9±0.5 8.8±1.4 7.7±0.9 5.0±1.2

Rainy 27±2.2 12.3±1.6 26.5±3.7 12.6±1.6 7.8±0.8 9.0±0.8 4.8±0.3

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Figure 4.7 Comparison of prey items consumed by different turtle species in different season (A) L. punctata (B) N. gangetica (C) P. smithii and (D) P. tecta.

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Table 4.4 Prey species richness (S), diversity Index (H') and evenness index (E) calculated during three different seasons of the year for each of the four turtle species in the Pothwar Plateau.

Seasons

Turtle Species Winter Summer Rainy

Lissemys punctata Prey species richness (S) 09 14 13

Diversity index (H') 2.2 2.6 2.6

Evenness index (E) 0.4 0.6 0.7

Nilssonia gangetica Prey species richness (S) 10 11 12

Diversity index (H') 2.3 2.4 2.5

Evenness index (E) 0.8 0.7 0.8

Pangshura smithii Prey species richness (S) 13 12 12

Diversity index (H') 2.6 2.5 2.5

Evenness index (E) 0.7 0.9 0.7

Pangshura tecta Prey species richness (S) 12 11 12

Diversity index (H') 2.5 2.3 2.5

Evenness index (E) 0.8 0.5 0.9

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16 14 Winter season summer season 16 Winter season summer season 14 13 14 Rainy season Rainy season 14 13 12 12 9 10 10 9 8 8

Seasons 6 Seasons 6 4 2.1 2.6 2.6 4 2.12.62.6 2 0.4 0.6 0.7 2 0.40.60.7 0 0 Prey species richness Diversity index Eveness index Prey species Diversity index Eveness index richness A Prey species richness, diversity and eveness index Prey species richness, diversity and eveness index B

14 16 14 Winter season summer season 12 12 Winter season summer season 14 13 Rainy season 12 11 Rainy season 12 10 9 10 8 8

6 Seasons

6 Seasons 4 2.1 2.6 2.6 4 2.52.42.5 2 0.4 0.6 0.7 2 0.80.50.9 0 0 Prey species richness Diversity index Eveness index Prey species Diversity index Eveness index richness C Prey species richness, diversity and eveness index D Prey species richness, diversity and eveness index

Figure 4.8 Prey species richness, diversity and evenness indices of four turtle species captured from Pothwar Plateau (A) Lissemys punctata (B) Nilssonia gangetica (C) Pangshura smithii (D) Pangshura tecta.

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summer, and rainy season) of stomach contents of L. punctata. Results showed higher consumed food item in gram was insects (25.5±3.3, 29±2.1 and 30.2±2.0) in every season and minimum was algae (Oscillatoria acuminate) (7.8±1.4, 9.8±2.0 and 7.8±0.3) respectively (Appendix VII). The mean consumption of different food items was highest in rainy season followed by summer and winter (Table 4.3, Fig.

4.7 A).

4.4.4.2 Indian softshell turtle (Nilssonia gangetica)

The food consumption of the soft shell turtle (N=33, 33 and 34) during three seasons (winter, summer and rainy) in stomach contents of N. gangetica was analyzed (AppendixVIII). Results showed that highest consumption of food item was insects (22.4±1.5, 24.3±1.1 and 31.2±0.8) and minimum was algae (8.9±0.9,

8.9±0.4 and9.8±1.1) respectively. The mean consumption of different food items was highest in rainy season followed by summer and winter (Table 4.3, Fig. 47 B).

4.4.4.3 Brown River turtle (Pangshura smithii)

In the three seasons viz, summer, winter and rainy (N= 36, 37 and 37) respectively analyzed (Appendix IX). The highest consumed food item was earthworms (29.5±0.3, 32.9±0.5 and 12.3±1.6) and lowest was algae (Table 4.3,

Fig. 4.7 C).

4.4.4.4 Indian roofed turtle (Pangshura tecta)

During three seasons stomach samples (N=16, 17 and 17, respectively) of

P. smithii were analyzed (AppendixX). The higest consumption of food items during different season was plant material (29.5±0.3, 20.9±0.5 and 12.6±1.6) and

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lowest was algae (Oscillatoria acuminate) (7.8±0.6, 8.8±1.4 and 7.8±0.8).

Mean consumption of food items in winter, summer and rainy season was statically compared by using two ways ANOVA. The consumption of prey items of three seasons was found to be significantly different (p < 0.05) in P. tecta while in rest of three species viz. L. punctata, N. gangetica and P. smithii it was non- significant (p > 0.05). (Appendix XI, XII, XIII, XIV).

4.4.5 Prey Species Richness (S), Diversity Index (H') and Evenness Index (E)

The diet composition of the four freshwater turtle species varied in three different seasons of the year, and this data was used to compute prey species richness (S), diversity Index (H') and evenness Index (E) of each turtle species studied (Table 4.4; Fig. 4.8).

Prey species Richness (S) for Lissemys punctata was found maximum (14) in summer season while least (09) in winter. Diversity Index (H') of prey species was high during summer and rainy season (2.6) but lowest least during winter season (2.2). The Evenness Index (E) of the prey species of Lissemys punctata was also high during summer and rainy seasons (0.6) as compared to winter season

(0.4) (Table 4.4; Fig.4.8A). For Nilssonia gangetica, prey species richness was maximum (12) during rainyseason but minimum (10) during winter season;

Diversity and evenness indices (2.5; 0.8, respectively) were also high during rainy season (Table 4.4; Fig. 4.8 B). For Pangshura smithii, prey species richness (13) and Diversity index (2.6) were high during winter season; however, evenness index was high during summer season (Table 4.4; Fig. 4.8 C). For Pangshura tecta, prey

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species richness and diversity index were high during winter and rainy seasons, whereas evenness index was found high during summer season (Table 4.4; Fig. 4.8

D).

4.4.6 Insects Orders Consumed by Freshwater Turtles

During current study, percent volume (% V) consumption of major insect’s orders by different turtle species from the study area revealed five different insect orders during the study period. The consumed insect’s orders included

Hymenoptera (29.5± 9.5), Coleoptera (28.5± 7.8), Diptera (23.5± 7.8), Homoptera

(18.2± 7.3) and Odonata (19.7± 4.3) (Table 4.5; Fig. 4.9). The Indian flap shell turtle (Lissemys punctata) consumed Hymenoptera and Coleoptera in equal percentage (40 %), followed by Diptera (36 %), Homoptera (25 %) and least

Odonata (22 %) (Table 4.5).

The Indian soft-shell turtle (Nilssonia gangetica) consumed Hymenoptera in highest percentage (50 %), followed by Coleoptera (43%) while least consumed was Odonata (30%). (Table 4.8). The Brown river turtle (Pangshura smithii) consumed Coleoptera in highest percentage (21 %), followed by Hymenoptera (20

%), and least Diptera (12 %). P. smithii were 17± 1.7 (Table 4.8). The Indian roofed turtle (Pangshura tecta) consumed only four insects’ orders including

Coleoptera in highest percentage (10.0 %), and Dipterain least (8 %) (Table 4.5;

Fig. 4.9).

A comparisons of insect orders consumed by four freshwater turtle species showed that Hymenoptera were most heavily consumed (29.5± 9.5 %), followed by

Coleoptera (28.5± 7.8 %), and Diptera (23.5± 7.8 %), while least consumed among

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five orders was Odonata (19.7± 4.3 %) (Table 4.5; Fig 4.9).

Insect orders and turtle species were compared through ANOVA, using

General Linear Model (GML SPSS-17). Analysis showed a significant difference

(F = 25.2df =3 and p< 0.05) between turtle species consumption of different insect orders, and non-significant difference among various insects orders consumed (F =

= 0.29df = 6 and p =0.87) (Appendix XV).

4.4.7 Plant Species Recorded from Turtles Species Stomach

Results showed that freshwater turtle species fed upon animal as well as plant matter. Determining the botanical composition of herbivore diet, micro- histological technique was applied. Analysis of 410 stomach flushed samples of four different freshwater turtle species of the study area revealed that they consumed five different plant species during the study period; redstem wormwood

(Artemisia scoparia), carrot Grass (Parthenium hysterophorus), ashwa gandha

(Withania somnifera), couch grass (Cynodon dactylon) and wild saff lower

(Carthamus oxycantha) (Fig. 4.10).

The Indian flap shell turtle (Lissemys punctata) consumed Parthenium hysterophorus in highest percent volume (11.3 %) followed by Artemisia scoparia

(10.2 %), while least consumed was Carthamus oxycantha (6.9 %). One average, this turtle species consumed 9.2 ± 1.0 % plant species, as a part of its overall diet

(Table 4.9; Fig. 4.10).

For Indian soft shell turtle (N. gangetica), plant material comprised of only

2.1 ± 0.8 %, on average (Table 4.9; Fig. 4.11). The plant species consumed in

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Table 4.5 Percent volume (% V) of insects orders recovered from the stomach

contents of freshwater turtle species (N=410) collected from selected study sites.

Species Hymenoptera Coleoptera Diptera Homoptera Odontata Mean± S.E L. punctata 40 40 36 25 22 32.6± 3.8 N. gangetica 50 43 38 34 30 39± 3.5 P. smithii 20 21 12 14 18 17± 1.7 P. tecta 8 10 8 0 9 7± 1.8 Mean± S.E 29.5± 9.5 28.5± 7.8 23.5± 7.8 18.2± 7.3 19.7± 4.3

Figure 4.9 Comparison of insect ordersconsumed by freshwater turtles of the

Pothwar Plateauvalues represent mean ± SE.

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Reference slides Sample slides

A A

B B

C C

D D

E E Figure 4.10 Photographs of reference and sample slides plant species prepared from recovered plant materials from the stomach contents offreshwater turtle species from the study area (A) Artemisia scoparia (B) Parthenium hysterophorus (C) Withania somnifera (D) Cynodon dactylon (E) Carthamus oxycantha.

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Table 4.6 Percent volume (% V) of plant species recovered from the stomach contents of freshwater turtle species (N=410) collected from the study area.

Species Artemisia Parthenium Withania Cynodon Carthamus Mean ± scoparia hysterophorus somnifera dactylon oxycantha S.E L. punctata 10.2 11.3 7.8 9.8 6.9 9.2 ± 1.0

N.gangetica 3.2 2.7 0 1.5 3.0 2.1 ± 0.8 P. smithii 9.6 10.9 10.7 8.2 8.6 9.6 ± 0.5 P. tecta 4.0 6.2 5.6 3.9 11.3 6.2 ± 1.4 Mean ± 8.0 ± 2.3 6.3 ±2.3 5.4 ± 1.8 6.6 ± 1.7 7.4 ± 1.7 S.E

Figure 4.11 Percent consumption of five different plant speciesby freshwater turtles in the study area.

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highest percentage volume was Artemisia scoparia (3.2 %), the other three plant species were consumed in minor percentages, while Withania somnifera was not in the diet of this turtle species.

For Brown river turtle (Pangshura smithii), the contribution of plant material in the diet ranged from 8.2 to 10.9 %, the mostly consumed plant species by this turtle species was Parthenium hysterophorus 10.9 %, followed by Withania somnifera 10.7 % (Table 4.9; Fig. 4.11).

For Indian roofed turtle (Pangshura tecta), the average plant material in the diet contributed 6.2 ± 1.4 % (Table 4.8) however Carthamus oxycantha was consumed inhighest percentage (11.3 %). On average, the four freshwater turtle species consumed Artemisia scoparia 8.0±2.3 %, Parthenium hysterophorus

6.3±2.3 %, Withania somnifera 5.4 ± 1.8%, Cynodon dactylon 6.6 ± 1.7 % and

Carthamus oxycantha 7.4 ± 1.7% (Table 4.9; Fig 4.11). Recorded plant species from stomach contents of turtle species were compared through ANOVA, using

General Linear Model (GML SPSS-17). There was a significant difference (F =

15.4 df =3 and p< 0.05) between turtle species, consumption different plant species, and non-significant difference between plant species (F = 0.18 df = 6 and p

=0.94) (Appendix XVI).

4.5 DISCUSSION

Food is a primary link between an animal and its environment. Chelonians play significant role in maintaining a healthy ecosystem and are considered as indicators of healthy aquatic ecosystem. They are the natural scavengersof aquatic ecosystem, thereby, cleaning water from dead organic detritus. Turtles feed on a

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variety of aquatic and semi-aquatic organisms’ viz. worms, insects, snails, slow movingaquatic crabs, dead animals and fragments of dead bodies, thus lowering the water pollution. Diets of freshwater turtles often reflect the availability of food resources in the environment also an opportunisticcarnivore that obtains its food from a wide variety of sources like plankton, nekton, benthic macro-organisms, carrion, and terrestrial organisms that fall upon the water.

There is scanty little literature available on the feedingecology of freshwater turtle species in Pakistan. In the current study, we applied stomach contents flushing method to investigate the diet composition of the freshwater turtle species in the Pothwar Plateau. Since the turtles are very shy, direct observations for feeding habits are veryrare. On the basis of analysis of stomach contents, the most preferred food items of freshwater turtles in the study area were found insects, earthworm, plants, snails and algae. Earlier on, Vijaya (1981) had reported similar findings that the freshwater turtles feed on earthworms and tadpoles. Lima et al.

(1997) concluded that turtles are omnivorous and feed in a strategic way on a greater quantity of benthonicmacro-invertebrates; insects, snails and crustaceans.

Among all stomach contents samples of the freshwater turtles analysed, insect body remains constituted highest percent frequency (%F) of occurrence 83.9

% (344) whereas the minimum % F was recorded for algae 25.6 % (105). Kennett and Tory (1996) agreed that most favorable food items of turtles are aquatic insects

(87.5 %) and less favorable were molluscaand gastropoda (8.8 %).

Results of the current study have indicated freshwater turtle species as omnivores because diverse types of food items were recovered from the stomach

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contents of the specimens captured. Stomach contents analysis revealed, on average insects body parts 26.3%, earthworm 19.9%, snails 16.1%, algae 11.4% and plants 10.2%. Whitaker andRomulus (1997) reported similar findings that turtles are omnivores feeding on vegetation as well as on animal flushes.

Auffenberg (1981) showed that turtles prey on living organisms, scavenge on organic detritus fromthe pond bottom and household garbage. One earlier published study (Bury, 1986) reported that freshwater turtles feed byvolume occurrence of prey items, insects 45.1 %, vegetation 0.8%, algae 24 %, crustacean

18.2 % and amphibian (Anura) 11.9 %.

The average consumption of food items in turtle species varied during different seasons, viz. winter, summer and rainy, in the study area. During rainy season maximum food items were consumed by different turtle species, followed by in summer season, but in winter season quantity of food items consumption was found lower as compared to other seasons. Luiselli et al. (2004) studied seasonal feeding habits of four freshwater turtle species and found significant difference in the frequency of consumption of prey items that were consumed more in the wet season but less in dry season by four species. The inter-seasonal differences in frequency of food items consumption was mainly due to the availability and preference of food items.

Five different insect orders consumed by the four freshwater turtle species in the current study included Hymenoptera, Coleoptera, Diptera, Homoptera and

Odonata. The most preferred insect order for turtle species during study period was hymenoptera whereas the least preferred was homoptera. Caputo and Vogt (2008) had reported that Testudines (Chelidae) feed on 14 different insect orders;

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Trichoptera, Diptera, Coleoptera, Cyclorrhapha, Hymenoptera, Blattodea,

Hemiptera, Orthoptera, Psocoptera, Ephemeroptera, Plecoptera, Collembola and

Arachnida. Similarly, Novelli et al. (2013) reported that turtles consume 16 categories of prey items, belonging to 16 orders and 70 families of Insects, including 3 families of Diplopoda, 2 of Crustacea, and 13 families of Arachnida.

Yamashita (1990) identified several orders of insects in feces of turtle species including Coleoptera, Isoptera, Hemiptera and Neuroptera.

Results of the current study have also indicated that freshwater turtle species of the study area are opportunistic omnivore’s thatconsume diversity of plant and animal items. Plant material recovered from the stomach contents showed that freshwater turtles consumed five different plant species; wormwood (Artemisia scoparia), carrot grass (Parthenium hysterophorus), ashwagandha (Withania somnifera), couch grass (Cynodon dactylon) and wild safflower Carthamus oxycantha. In a similar studyBrock (1988) had investigated that plant material is important component of turtle’s diet. Similar findings related to foraging strategy among Emydidae species, were recorded by Caputo and Vogt (2008) who concluded that turtles feed on any available prey item in their habitat. Most preferable food items of family Chelidae included different stages of plant material

(seed, fruit, and leaves) including seven different plant families; Arecaceae (seed)

Rubiaceae seed, Leguminosae fruit, Rodophyta, Batrachospermum, Guttiferae fruit and Rodophyta.

4.6 CHAPTER SUMMARY

The diet composition of freshwater turtle species inhabiting the Pothwar

Plateau was investigated by using stomach contents flushing method. Stomach

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contents samples of N=410 specimens of four different turtle species were collected from the study area, and ﬁxed in 70% ethanol till final analysis. During analysis, each sample was segregated into different categories including animal matter (insects and other invertebrates), plant matter (like vegetation including algae), and other materials. Contents were identified up to the lowest possible taxa.

Analysis of stomach contents of freshwater turtles revealed five different kinds of prey items including insects, earthworms, snails, plants, and algae, in addition to soil particles and some unidentified material. The turtles were found omnivorous in their feeding behavior, but highly opportunistic, feeding on any availableprey item in the environment. Mean percent volume (% V) of prey items recovered from stomach contents included insects 26.3 %, earthworms 19.9 %, snails 16.1%, plants 10.2 %, and algae 11.4 %, besides soil particles 7.5 % and unidentified material 8.4 %.

The diet composition of freshwater turtles varied among different seasons; during rainy and summer seasons, there was greater consumption of insects

(30.2%, and 45.3%, respectively) as compared to the winter season (20.2%).

Similarly, earthworms were consumed in greater percentage in summer (20.2%) and rainy seasons (29.9%) than in winter (17.2 %). Snails were consumed heavily in summer (15.8%) and rainy (24.4%) seasons. The plant material was also consumed in maximum percentage in rainy season (12.1%), followed by summer season (9.5%), but least in winter (5.5%).

The insect consumed by freshwater turtles were identified belonging to five different orders including Hymenoptera (29.5 ± 9.5 %) Coleoptera (28.5 ± 7.8%),

Diptera (23.5 ± 7.8%), Homoptera (18.2 ± 7.3%) and Odonata (19.7 ± 4.3%).

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Similarly, five different species of plants were consumed on regular basis by freshwater turtles studied; average consumption included Artemisia scoparia (8.0 ±

2.3%), Parthenium hysterophorus (6.3 ± 2.3 %), Withania somnifera (5.4 ± 1.8 %),

Cynodon dactylon (6.6 ± 1.7 %) and Carthamus oxycantha (7.4 ± 1.7 %).

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Chapter 5

GENERAL DISCUSSION

Freshwater turtles are a metamorphic primitive and ecologically important group with long and compound biography in a variety of aquatic ecosystem (Bour, 2008).They have vital role in aquatic habitat similar to vultures which they play as scavengers in terrestrial ecosystem (TCF, 2002). They often participate as fundamental species keystone for other animals and plants, and show major importance for environments. They serve as web of interacting and co- dependent species that constitute a healthy functioning of the ecosystem. However, freshwater turtles are a group of high conservation concern, with approximately

40% of the world’s turtle species currently listed as globally threatened by IUCN

Rhodin et al. (2010). It is estimated that freshwater turtles are being traded atnational and international levels worth billions of US dollars ($) annually (Roe and Georges, 2008; Smith et al., 2009). Demand of turtle specimens for various purposes includes the markets for food, luxury goods, tourist curios, medicines and live animals for pet trade (Nijman, 2010). Resultantly, many species of freshwater turtles have become threatened. The freshwater turtle species including Nilssonia gangetica, Nilssonia hurum and Geoclemys hamiltoni have been included in

Appendix-I, whereas, Lissemys punctata is in Appendix-II of the Convention on

International Trade in Endangered Species (CITES) (Anonymous, 2005). It is important to mention that several countries of the Asian region have not been up to the mark and made just few attempts for conservation and management of freshwater turtle resource.

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Pakistan, a part of Indian subcontinent, has a variety of flora and fauna in different areas basically due to diversity in its landscape and bioclimatic zones in its different regions (Khan, 1980; Mufti et al. 1997). In Pakistan, freshwater turtles are widely distributed, and some check lists are also available about various species in different parts of the country. However, the distribution and abundance of freshwater turtle species in the Pothwar Plateau was unexplored largely. In the current study, the two main factors that restricted the distribution and occurrence of freshwater turtle species in the study area are water bodies and availability of food.There were also significant relationships observed distribution of turtles from the study area and characteristics of the habitat. The current study has revealed four species of freshwater turtles, distributed in the water bodies of the Pothwar Plateau;

Indian flap shelled turtle (Lissemys punctata), Indian soft shell turtle (Nilssonia gangetica), Brown river turtle (Pangshura smithii) and Indian roofed turtle

(Pangshura tecta), with the species Lisseymes punctata occurring predominantly in the study area. In Pakistan, two families of freshwater turtles, namely,

Geoemydidae and Trionychidae occur in the rivers, ponds and other water bodies, the two families includingeight different freshwater turtle’s species(Anonymous,

2005). Earlier on, Akbar et al. (2006) reported eight freshwater turtle species including Brown River Turtle (Pangshura smithii), Indian Saw-backed turtle

(Pangshura tecta), Indian Soft-shell Turtle (Nilssonia gangetica), and Indian Flap- shell Turtle (Lissemys punctata), in addition to four other species. Similarly,

Mahmood et al. (2012) had reported three species of freshwater turtles distributed in Korang River, Rawalpindi/Islamabad, including Indusmud turtle (Lissemys punctata), the Indian soft shell turtle (Nilssonia gangetica) and the Brown river

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turtle (Pangshura smithii). But the current study has shown four species of freshwater turtles distributed in the Pothwar Plateau. Azam et al. (2005) reported six species of freshwater turtles including Pangshura smithii, Pangshura tecta,

Hardella thurgi, Chitra indica, Nilssonia gangatica and Lissemys punctata occurringin the River Indus, adjacent to Sukktar Barrage, Guddu Barrage,

JamaldinWali and pond areas adjacent to Kandhkot. The Chitra indica and

Pangshura smithii were found to be abundant in various parts of the River Indus

Azam et al. (2005). Williams (1980) had shown that turtle distribution and abundance is influenced by availability of potential food and approach of that area, and if resources are restricted eventually through habitat degradation, then reproduction success suffered badly and population of turtles may decline.

The freshwater turtles are present all over the world including the Asian countries; Pakistan, India, Nepal, Bangladesh and Sri Lanka, however, still being data deficient, especially regarding their population estimates Buhlmann et al.

(2009). The studies on population estimates of freshwater turtles are lacking not only in Pakistan, but in South Asia as well. They are rarely observed due to several reasons such as change in habitat, food resources and seasonal incline Donner

Wright et al. (1999). Population status of freshwater turtle species estimated in

Pothwar Plateau; present study results showed a total of N= 3004 and N=1681 individual turtles (by CMR, and direct count method, respectively), belonging to two families viz., Trionychidae (Lissemys punctata and Nilssonia gangetica) and

Geoemydidae (Pangshura smithii and Pangshura tecta) were recorded from all sixteen selected sampling sites. Four different species were recorded in water bodies of the district Rawalpindi whereas only two species were found in district

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Chakwal. Both methods used revealed Lissemys punctataas most abundant turtle species, followed by Pangshura smithii, and Nilssonia gangetica while Pangshura tectawas found least abundant. Some earlier published studies are available about the turtle species occurring in the country. For example, Khan (2015) observed six freshwater turtle species in district Thatta of Sindh province; among those Lissemys punctata and Pangshura smithii, were found abundant. Chitra indica, Nilssonia gangetica and Nilssonia hurum were common, Hardella thurjii was less common.

Similarly, Khan et al.(2015) reported four species of freshwater turtles from district

Badin (Sindh); Lissemys punctata (37 %) and Chitra indica(40%), were abundant,

Nilssonia hurum (18%) was common while Geoclemys hamiltonii (4.31%), was found rare.

In the current study, density estimates of freshwater turtles have shown an overall of 92% variation during winter/fall season (in comparison summer /spring season) in the Pothwar Plateau during two years of study period (2012-2014). This may be attributed to various factors including mainly hibernation, insufficient food items cold weather, illegal capture and kill, professional hunters and local people for getting their shell to sell in the black market for which various reports exist.

Hibernation is an established fact in turtle’s life history in cold winter weather during annual cycle of temperate-zone turtles. Ultsch and Jackson (1982) showed that during hibernation condition turtles restricted their movement and activities.

Kayani et al. (2015 have reported some general body measurements of freshwater turtle species; in the current study, different age classes (juveniles and adults) have been focused for their morphometric studies. During the current study,

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a total of N=1681 specimens of four different freshwater turtle species were collected from the different sampling sites of the study area for their morphometrical analysis. Recorded body measurements showed Nilssonia gangetica being the largest and heaviest turtle species with maximum body weight

(4300± 6.5 g) and size among all four turtle species recordedin the study area.

There was sexual dimorphism between males and females of this species; females weighed on average 4300 ± 6.5g while males weighed 3500± 6.1 g.

Somewhat similar morphological characteristics were described by Mahmood et al.

(2012) in three freshwater turtle species Lissmeys punctata, Nilssonia gangetica and Pangshura smithii from Rawalpindi and Islamabad area. According to their findings body weight of Nilssonia gangetica (3045.9g) as well as size of its carapace and plastron was found maximum among the three turtle species studied.

The populations of turtles in a habitat depend upon different factors, most importantly the availability of food sources and ability of turtles to access that food source. Population of turtles may decrease in a particular habitat if recourses are limited due to overuse of recourses or due to modification of local habitat. The scarcity of food may also affect reproductive potential of turtles, their growth rate and survival rate due to increased mortality, which ultimately may result in decline in population. For diet analysis of the freshwater turtle species, during current study, a total of (N=410) stomach content samples of four species of freshwater turtles were collected from different sampling sites and analysed for investigating their feeding habits. Results showed freshwater turtles having an omnivorous feeding behavior, but in all species, the most preferred and most frequently

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consumed prey items were insects (26.3 %). Among insects, the preferred insect orders included Hymenoptera Coleoptera, Diptera, Homoptera and Odonata. The prey items of the four turtle species studied were found similar.

During different seasons of the year including; winter, summer and rainy, during study period food consumption in freshwater turtles was variable; maximum food items were consumed during rainy season followed by summer and winter. In

Lissemys punctata and Nilssonia gangetica most preferred food items during rainy season were insects (30.2±2.0 %) while algae (Oscillatoria acuminate) were less preferred (7.8±0.3 %. In Pangshura smithii earthworms were heavily consumed

(29.5±0.3 %) while lowest were algae (8.8±0.8 %). In Pangshura tecta, during rainy season, highestly consumed food items were plants (29.5±0.3%) and lowest

(7.8±0.6) was algae (Oscillatoria acuminate). Hossain et al. (2012) reported seasonal variation in diet consumption of Bellamaya species. The mean consumption of different food items in summer, rainy and winter season was (57.2

± 34.6, 86.4 ± 54.4 and 28.16 ± 19.5%), respectively. Kennett and Tory (1996) described dietary preference in two freshwater turtle species; Chelodina rugosa and

Elseya dentata during wet and dry seasons. According to their findings composition of food items during wet, and dry seasons were similar. But percentage of food items consumed by the two species was found more (69 %) in wet season as compared to dry season (31%).

Results of the current study have indicated that most preferred food items of all four species of freshwater turtle during the study period. All four species consumed five different insect orders viz. Hymenoptera, Coleoptera, Diptera,

Homoptera and Odonata. The most preferred insect order was found to be

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hymenoptera while the least preferred was homoptera. Allanson and Georges

(1999) had reported similar findings in two species of freshwater turtles (Elseya purvisi and Elseya georges) from Australia whereby they were found extensively insectivorous, mostly consuming insect orders; Coleoptera, Diptera, Herniptera,

Odonata and Plecoptera.

In the current study, all four freshwater turtle species also consumed five different plant species; wormwood (Artemisia scoparia), carrot grass (Parthenium hysterophorus), ashwagandha (Withania somnifera), couch grass (Cynodon dactylon) and wild safflower Carthamus oxycantha. Brock (1988) had investigated and concluded that plant material was an important component of turtle’s diet.

Similar findings related to foraging strategy among Emydidae species, were recorded by Caputo and Vogt (2008), who reported that turtles feed on any available prey item in their habitat. Most preferable food items of family Chelidae included different stages of plant material (seed, fruit, and leaves) including seven different plant families; Arecaceae (seed) Rubiaceae seed, Leguminosae fruit,

Rodophyta, Batrachospermum, Guttiferae fruit and Rodophyta.

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SUMMARY

Freshwater turtles form important biodiversity component of the aquatic ecosystems and have much significance due to their ecological position and economic value. They are generally covered with a protective shell, a special bony or cartilaginous, one of the most enduring morphological metamorphoses in vertebrate history. However, the shell also imposes biggest threat to turtles populations since it is used as ingredients in the Traditional Chinese Medicines

(TCM), ornamental value, and also utilized in making tools and musical instruments. Freshwater turtles are also an important food source for human in some Asian countries. Therefore, the turtle species are illegally captured and killed for obtaining their shell, which are then traded across the border to other countries in Asia including China. In Pakistan, scientific studies focusing on ecological parameters and habitat requirements of freshwater turtles have been scanty although a few have reported checklists and their distribution in some parts of the country. The current study, therefore, was designed to investigate the distribution, populations, and food habits of different freshwater turtle species inhabiting the

Pothwar Plateau.

The distribution of freshwater turtle species was studied by conducting surveys of the water bodies in the study area, and recording visual observationsalong the water body (250 m on either side of the one kilometer transect). In addition, cast net was operated and used to capture different turtle species to confirm their species identification and distribution. Population estimation of freshwater turtles was carried out by using “capture, mark and

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recapture (CMR)” method and also direct count methods while their food habits were investigated by analysis of stomach contents flushing.

Results showed four freshwater turtle species found distributed in water bodies throughout the districtRawalpindi including Indian flap shell turtle

(Lissemys punctata), Indian soft shell turtles (Nilssonia gangetica), The Indian roofed turtle (Pangshura tecta), and brown river turtle (Pangshura smithii). The

Lissemys punctata was found most abundant, Nilssonia gangetica and Pangshura smithii were common whereas Pangshura tecta was least common. While only two species (Lissemys punctata and Panshura smithii) occurred in the water bodies of the other district (Chakwal), the remaining two species being absent. In the whole study area, Lissemys punctata occurred at 15 out of 16 total sampling water bodies,

N. gangetica at 10 sites, P. smithii at 11 sites and P. tecta at only 5 sites.

A total of N = 2166 specimens of freshwater turtles were captured using

CMR method; Lissemys punctata was found most abundant species (48.1%), followed by Pangshura smithii (25.2%), and Nilssonia gangetica (20.2%) while the least common was Pangshura tecta (6.2%)A total of 3004 specimens of turtles were counted during the study period using direct count method; again Lissemys punctata was the most abundant species (41.8%), followed by Pangshura smithii

(30.4%), and Nilssonia gangetica (19.1%) whereas Pangshura tecta (8.5%)was foundleast common. Estimates of average population density (per square kilometer) of freshwater turtles were (20.8/ Km²) by direct count method as compared to CMR method during study period.

Average population density offreshwater turtles recorded by direct count

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method (20.8/ Km²) was higher as compared to the CMR method (15.0/Km²) used.

The estimated density of Lissemys punctat was 104.6/Km², Nilssonia gangetica

47.9/Km², Pangshura smithii 76.2/Km² and Pangshura tecta 2 1.5/Km²during the study period from 2012-2014. Seasonal populations (summer and winter) of freshwater turtle species were found significantly different; in summer/spring season, turtle populations were found high while during winter/fall season, turtlespopulations were found low.

For investigating diet composition of freshwater turtle species, a total of four hundred and ten (N=410) stomach contents samples of four different species of freshwater turtles were collected from different sampling sites and analysed.

Results showed that all four turtle species studied were found omnivorous in their feeding behavior but mainly consuming animal and plant matters. From stomach contents of all four turtle species insects and earthworms were recovered in large proportions (26.3 % and 19.9 %, respectively) as compared to plant parts and algae

(10.2 % and 11.4 %, respectively).

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LITERATURE CITED

Akbar, M., M. Mushtaq-ul-Hussain and Zaib- ul- Nisa. 2006. Distribution of fresh

water turtles in Punjab, Pakistan. Caspian J. Env. Sci., 4: 142-146.

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APPENDICES

Appendix- I Statistical comparison of (Lissemys punctata) populations by using two methods (CMR and direct count) at sixteen selected sampling sites of the

Pothowar Plateau by Tukey’s Post-Hoc test.

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

2 NS

3 NS NS

4 NS NS NS

5 NS NS NS NS

6 NS NS NS NS NS

7 NS NS NS NS NS NS

8 NS NS NS NS NS NS NS

9 NS NS NS NS NS NS NS NS

10 NS NS NS NS NS NS NS NS NS

11 S S S NS NS NS NS NS NS NS

12 S S S NS NS NS NS NS NS NS NS

13 S S S NS NS NS NS NS NS NS NS NS

14 S S S NS NS NS NS NS NS NS NS NS NS

15 S S S NS NS NS NS NS NS NS NS NS NS NS

16 S S S NS NS NS NS NS NS NS NS NS NS NS NS

* NS=Non significant difference of (Lissemys punctata) population among selected sites. * S= Significant difference of (Lissemys punctata) population among selected sites.

172

Appendix- II Statistical comparison of (Nilssonia gangetica) populations by using two methods (CMR and direct count) at sixteen selected sampling sites of the

Pothowar Plateau by Tukey’s Post-Hoc test.

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

2 NS

3 NS NS

4 NS NS NS

5 NS NS NS NS

6 NS NS NS NS NS

7 NS NS NS NS NS NS

8 NS NS NS NS NS NS NS

9 NS NS NS NS NS NS NS NS

10 NS NS NS NS NS NS NS NS NS

11 S NS NS NS NS NS NS NS NS NS

12 S NS NS NS NS NS NS NS NS NS NS

13 S NS NS NS NS NS NS NS NS NS NS NS

14 S S NS NS NS NS NS NS NS NS NS NS NS

15 S S S NS NS NS NS NS NS NS NS NS NS NS

16 S S S S S S S S S S S S S NS NS

* NS=Non significant difference of (Nilssonia gangetica) population among selected sites. * S= Significant difference of (Nilssonia gangetica) population among selected sites.

173

Appendix- III Statistical comparison of (Pangshura smithii) populations by using two methods (CMR and direct count) at sixteen selected sampling sites of the Pothowar Plateau by Tukey’s Post-Hoc test.

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

2 NS

3 NS NS

4 NS NS NS

5 NS NS NS NS

6 NS NS NS NS NS

7 NS NS NS NS NS NS

8 NS NS NS NS NS NS NS

9 NS NS NS NS NS NS NS NS

10 NS NS NS NS NS NS NS NS NS

11 NS NS NS NS NS NS NS NS NS NS

12 NS NS NS NS NS NS NS NS NS NS NS

13 NS NS NS NS NS NS NS NS NS NS NS NS

14 NS NS NS NS NS NS NS NS NS NS NS NS NS

15 NS NS NS NS NS NS NS NS NS NS NS NS NS NS

16 S S NS NS NS NS NS NS NS NS NS NS NS NS NS

* NS=Non significant difference of (Pangshura smithii) population among selected sites. * S= Significant difference of (Pangshura smithii) population among selected sites.

174

Appendix- IV Statistical comparison of (Pangshura tecta) populations by using two methods (CMR and direct count) at sixteen selected sampling sites of the Pothowar Plateau by Tukey’s Post-Hoc test.

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

2 NS

3 NS NS

4 S NS NS

5 S NS NS NS

6 S NS NS NS NS

7 S NS NS NS NS NS

8 S NS NS NS NS NS NS

9 S NS NS NS NS NS NS NS

10 S NS NS NS NS NS NS NS NS

11 S S NS NS NS NS NS NS NS NS

12 S S NS NS NS NS NS NS NS NS NS

13 S S NS NS NS NS NS NS NS NS NS NS

14 S S S S S S S S NS NS NS NS NS

15 S S S S S S S S S S NS NS NS NS

16 S S S S S S S S S S S S S S S

* NS=Non significant difference of (Pangshura tecta) population among selected sites. * S= Significant difference of (Pangshura tecta) population among selected sites.

175

Appendix-V Two way analysis of variance (ANOVA) prey items consumption and different four turtle species.

Source of variation df SS MS F P- value

Prey items 6 14635.0 2439.2 7.8907 < 0.05

Species 3 10223.9 3411.0 11.0344 < 0.05

Residuals 1 85564.1 309.1

Appendix-VI Two way analysis of variance (ANOVA) prey items consumption and different locations selected during study period.

Source of variation df SS MS F P- value

Locations 14 5423.1 387.36 17.2 < 0.05

Prey items 6 3950.4 658.4 29.3 <0.05

Residuals 84 1886.8 22.4

176

Appendix- VII Seasonal variation of food consumption of Lissemys punctata collected from selected sampling sites of Pothwar Plateau.

Seasons Months Insects Earthworms Snails Plant material Algae Soil particles Unidentified material November 35.6 25.7 22.8 14.8 10.5 9.8 8.8 Winter December 25.6 22.9 20.9 10 9.8 7.8 7.8 Season January 22.8 19.8 19.4 9.8 6.3 5.3 5.6 (N=50) February 18 22.8 15.7 9.8 4.6 3.1 4.2 Mean ± S.E 25.5±3.3 22.8± 1.4 19.7±1.5 11.1±1.2 7.8± 1.4 6.5±1.4 6.6±1.0 March 33.8 29.8 23.4 13.5 13.4 6.5 4.5 Summer April 29.9 25.6 19.7 12.7 12.4 5.5 3.5 Season May 28.7 18.9 18.9 9.1 9.1 5.5 3.5 (N=50) June 23.6 17.7 17.2 4.3 4.3 2.5 2.5 Mean ± S.E 29±2.1 23±2.8 19.8± 1.3 9.9± 2.0 9.8±2.0 5±0.8 3.5±0.4 July 30.6 29.7 19.9 9.8 8.7 5.8 7.6 Rainy Season August 25.7 21.5 19.2 7.6 7.6 4.9 5.9 (N=50) September 21.1 22.2 15.8 9.6 7.4 4.3 6.5 October 22.6 22.6 14.3 9.8 7.5 5.8 5.2 Mean ± S.E 30.2±2.0 24±1.9 17.3±1.3 9.2±0.5 7.8±0.3 5.2±0.3 6.3±0.5

177

Appendix- VIII Seasonal variation of food consumption of Nilssonia gangetica collected from selected sampling sites of Pothwar Plateau.

Seasons Months Insects Earthworms Snails Plant Algae Soil particles Unidentified material material

November 19.8 30.3 18.1 11.6 9.8 5.6 7.2 Winter December 19.6 29.8 12.7 13.9 7.5 7.5 5.2 Season January 20.1 28.7 11.3 12.5 6.7 6.7 6.3 (N=36) February 19.7 29.2 28.3 12.4 7.2 5.8 6.5 Mean ± S.E 19.8±0.1 29.5±0.3 17.6 ±3.8 12.6±0.4 7.8±0.6 6.4±0.4 6.3±0.4 March 22.3 32 12.8 10.8 9.2 4.2 5.3 Summer April 20.9 31.8 12.5 9.6 7.1 3.1 7.9 Season May 19.7 33.9 13.8 15.9 12.6 8.7 9.7 (N=37) June 20.7 33.9 10.9 14.1 6.3 4 7.9 Mean ± S.E 20.9±0.5 32.9±0.5 12.5±0.6 12.6±1.4 8.8±1.4 5±1.2 7.7±0.9 July 7.9 7.9 19.7 21.5 10.9 9.9 1.5 Rainy August 13.8 13.7 33.7 32.1 6.9 7.9 0.3 Season September 12.9 11.7 32.9 29.1 8.2 9.3 0.5 (N=37) October 15.8 15.9 20.9 26.1 9.2 8.1 1.7 Mean ± S.E 12.6±1.6 12.3±1.6 26.8±3.7 27.2±2.2 8.8±0.8 8.8±0.8 4±0.3

178

Appendix-IX Seasonal variation of food consumption of Pangshura smithii collected from selected sampling sites of Pothwar Plateau.

Seasons Months Insects Earthworms Snails Plant Algae Soil particles Unidentified material material November 26.9 21.7 19.1 12.8 10.8 11.8 7.6 Winter December 21.9 21.9 17.3 11.5 7.6 8.9 6.5 Season January 20.9 19.3 18.5 11.6 6.9 8.6 5.9 (N=33) February 19.9 19.9 19.9 14.9 10.3 10.3 6.8 Mean ± S.E 22.4±1.5 20.7±0.6 18.7±0.5 12.7±0.7 8.9±0.9 9.9±0.7 6.7±0.3 March 21.4 19.8 19.8 11.5 8.7 10.7 7.8 Summer April 25.8 17.6 17.6 11.8 9.6 9.6 5.7 Season May 23.5 20.3 20.3 12.7 9.7 9.6 6.6 (N=33) June 26.5 25.9 25.9 10.8 7.6 9.7 6.7 Mean ± S.E 24.3±1.1 20.9 17.6±1.7 11.7±0.3 8.9±0.4 9.9±0.2 6.7±0.4 July 32.5 23.7 19.8 13.9 11.9 6.5 6.5 Rainy August 32.8 23.5 14.7 13.9 10.8 3.4 4.4 Season September 29.8 19.8 15.4 14.1 9.8 3.9 6.9 (N=34) October 29.7 18.6 11.7 11.7 6.7 1.8 1.8 Mean ± S.E 31.2±0.8 21.4±1.2 15.4±1.6 13.4±0.5 9.8±1.1 3.9±0.9 4.9±1.1

179

Appendix-X Seasonal variation of food consumption of Pangshura tecta collected from selected sampling sites of Pothwar Plateau

Seasons Months Insects Earthworms Snails Plant Algae Soil particles Unidentified material material

November 19.8 11.6 18.1 30.3 9.8 7.2 5.6 Winter December 19.6 13.9 12.7 29.8 7.5 5.2 7.5 Season January (N=16) 20.1 12.5 11.3 28.7 6.7 6.3 6.7 February 19.7 12.4 28.3 29.2 7.2 6.5 5.8 Mean ± S.E 19.8±0.1 12.6±0.4 17.6 ±3.8 29.5±0.3 7.8±0.6 6.3±0.4 6.4±0.4 March 10.8 32 12.8 22.3 9.2 5.3 4.2 Summer April 9.6 31.8 12.5 20.9 7.1 7.9 3.1 Season May (N=17) 15.9 33.9 13.8 19.7 12.6 9.7 8.7 June 14.1 33.9 10.9 20.7 6.3 7.9 4 Mean ± S.E 12.6±1.4 32.9±0.5 12.5±0.6 20.9±0.5 8.8±1.4 7.7±0.9 5±1.2 July 21.5 7.9 19.7 7.9 9.2 10.5 6.5 Rainy August 32.1 13.7 33.7 13.8 7.4 9.8 3.2 Season September (N=17) 29.1 11.7 32.9 12.9 7.7 8.8 4.6 October 26.1 15.9 20.9 15.8 6.9 6.9 4.9 Mean ± S.E 27±2.2 12.3±1.6 26.5±3.7 12.6±1.6 7.8±0.8 9.0±0.8 4.8±0.3

180

Appendix- XI Mean consumption of food items in (Lissemys punctata) during different season was statically compared by using two -ways ANOVA.