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The Feeding Behaviour and General Histological Characteristics of the Gastrointestinal Tract of South African Cave-Dwelling Amphipods

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The Feeding Behaviour and General Histological Characteristics of the Gastrointestinal Tract of South African Cave-Dwelling Amphipods THE FEEDING BEHAVIOUR AND GENERAL HISTOLOGICAL CHARACTERISTICS OF THE GASTROINTESTINAL TRACT OF SOUTH AFRICAN CAVE-DWELLING AMPHIPODS SIMONE VAN TONDER DISSERTATION SUBMITTED IN FUFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF MAGISTER SCIENTIAE in BIODIVERSITY AND CONSERVATION in the FACULTY OF SCIENCE at the UNIVERSITY OF JOHANNESBURG Supervisor: Dr J.F. Durand NOVEMBER 2006 DECLARATION DECLARATION I declare that this dissertation hereby submitted by me, Simone van Tonder, to the University of Johannesburg for the degree of Magister Scientiae in Biodiversity and Conservation has not been submitted by me for a degree at another university and that this is my own original work. Simone van Tonder Department of Zoology The University of Johannesburg P.O. Box 524 Auckland Park Johannesburg South Africa ii TABLE OF CONTENTS TABLE OF CONTENTS DECLARATION .................................................................................................... ii TABLE OF CONTENTS....................................................................................... iii LIST OF FIGURES ............................................................................................ ..vi LIST OF TABLES..................................................................................................x ACKNOWLEDGEMENTS ...................................................................................xiii ABSTRACT.................................................................................................. xiv OPSOMMING………………………………………………………………………xvi CHAPTER 1....................................................................................................... xvi GENERAL INTRODUCTION.............................................................................1 1.1 INTRODUCTION .....................................................................................1 1.2 MOTIVATION AND PROBLEM STATEMENT.........................................4 1.3 AIMS OF THE STUDY.............................................................................6 CHAPTER 2..........................................................................................................8 LITERATURE REVIEW.....................................................................................8 CAVE AND KARST SYSTEMS .....................................................................8 AMPHIPOD BIOLOGY ................................................................................11 AMPHIPOD FEEDING.................................................................................14 THREATS TO AMPHIPODS........................................................................19 CHAPTER 3........................................................................................................23 MATERIALS AND METHODS.........................................................................23 3.1 FIELD STUDIES ....................................................................................23 3.2 ENVIRONMENTAL ROOM STUDIES ...................................................26 3.3 STUDIES OF THE MOUTHPARTS OF SOUTH AFRICAN CAVE- DWELLING AMPHIPODS...................................................................28 3.4 HISTOLOGY OF THE GASTROINTESTINAL TRACT (GIT), AND THE USE OF HISTOCHEMISTRY IN DETERMINING THE GUT CONTENTS OF AMPHIPODS............................................................29 3.4.1 General morphology and orientation of the GIT was established using histology..............................................................................................29 3.4.2 The use of histochemistry in determining the constituents of the GIT contents ..............................................................................................31 3.5 FEEDING BEHAVIOUR OF SOUTH AFRICAN CAVE-DWELLING AMPHIPODS ......................................................................................34 3.5.1 Starvation of amphipods ....................................................................38 3.5.2 Bat faeces, leaf litter, sediment, and yeast as food sources for amphipods ..........................................................................................39 3.5.3 0.04g of bat faeces, 0.04g of leaf litter, 0.04g of sediment, and 0.04g of yeast as a food source for amphipods over a period of 36 days.....40 3.5.4 Twenty four hour study ......................................................................42 3.5.5 Extraction of amphipod gut contents, to determine if bat faeces, leaf litter, sediment, and yeast were ingested............................................43 iii TABLE OF CONTENTS 3.5.6 Predation, cannibalism, and scavenging in amphipods .....................44 3.6 THE USE OF MICROBIOLOGY IN DETERMINING THE CONSTITUENTS OF THE GUT CONTENTS OF CAVE-DWELLING AMPHIPODS ......................................................................................46 3.6.1 Heterotrophic (Standard) Plate Count (Pour plate method) ...............48 3.6.2 Total Coliforms (Membrane filtration procedure)................................50 3.6.3 Faecal Coliforms (Membrane filtration procedure).............................52 3.6.4 Confirmatory test for Escherichia coli ................................................54 3.6.5 Faecal Streptococci/Enterococci (Membrane filtration procedure).....55 3.6.6 Clostridium (Membrane filtration procedure)......................................57 3.6.7 Pseudomonas (Membrane filtration procedure).................................59 3.6.8 Detection of Salmonella spp. (Presence / Absence)..........................61 CHAPTER 4........................................................................................................64 RESULTS........................................................................................................64 4.1 FIELD STUDIES ....................................................................................64 4.2 ENVIRONMENTAL ROOM STUDIES ...................................................66 4.3 STUDIES OF THE MOUTHPARTS OF SOUTH AFRICAN CAVE- DWELLING AMPHIPODS...................................................................67 4.4 HISTOLOGY OF THE GASTROINTESTINAL TRACT (GIT), AND THE USE OF HISTOCHEMISTRY IN DETERMINING THE GUT CONTENTS OF AMPHIPODS............................................................76 4.4.1 General morphology and orientation of the GIT was established using histology..............................................................................................76 4.4.2 The use of histochemistry in determining the constituents of the GIT contents ..............................................................................................82 4.5 FEEDING BEHAVIOUR OF SOUTH AFRICAN CAVE-DWELLING AMPHIPODS ......................................................................................87 4.5.1 Starvation of amphipods ....................................................................94 4.5.2 Bat faeces, leaf litter, sediment, and yeast as a food source for amphipods ..........................................................................................95 4.5.3 0.04g of bat faeces, 0.04g of leaf litter, 0.04g of sediment, and 0.04g of yeast as a food source for amphipods over a period of 36 days...103 4.5.4.1 Twenty four hour study .................................................................104 4.5.4.2 Forty eight hour study ...................................................................105 4.5.4.3 Seventy two hour study.................................................................106 4.5.5 Extraction of amphipod gut contents, to determine if bat faeces, leaf litter, sediment, and yeast were ingested..........................................107 4.5.6 Predation, cannibalism, and scavenging in amphipods. ..................114 4.6 THE USE OF MICROBIOLOGY IN DETERMINING THE CONSTITUENTS OF THE GUT CONTENTS OF CAVE-DWELLING AMPHIPODS ....................................................................................120 4.6.1 Heterotrophic (Standard) Plate Count (Pour plate method) .............120 4.6.2 Total coliforms (Membrane filtration procedure) ..............................125 4.6.3 Faecal Coliforms (Membrane filtration procedure)...........................130 4.6.4 Confirmatory test for Escherichia coli ..............................................135 4.6.5 Faecal Streptococci/Enterococci (Membrane filtration procedure)...136 iv TABLE OF CONTENTS 4.6.6 Clostridium (Membrane filtration procedure)....................................141 4.6.7 Pseudomonas (Membrane filtration procedure)...............................145 3.6.8 Detection of Salmonella spp. (Presence / Absence)........................150 CHAPTER 5......................................................................................................155 DISCUSSION................................................................................................155 5.1 FIELD STUDIES ..................................................................................155 5.2 ENVIRONMENTAL ROOM STUDIES .................................................157 5.3 STUDIES OF THE MOUTHPARTS OF SOUTH AFRICAN CAVE- DWELLING AMPHIPODS.................................................................158 5.4 HISTOLOGY OF THE GASTROINTESTINAL TRACT (GIT), AND THE USE OF HISTOCHEMISTRY IN DETERMINING THE GUT CONTENTS OF AMPHIPODS..........................................................158 5.4.1 General morphology and orientation of the GIT was established using histology............................................................................................158
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