ECTOPARASITE FAUNA OF RODENTS AND SCANDENTS IN . … fauna of rodents and scandents... · granulatus, Laelaps nuttalli, Leptotrombidium sp., Leptotrombidium deliense, Sarcoptes scabiei,

ECTOPARASITE FAUNA OF RODENTS AND SCANDENTS IN DIFFERENT ECOLOGICAL HABITATS AND ITS POTENTIAL

ZOONOTIC IMPLICATIONS IN MALAYSIA

Madinah Blnti Adrus

Master of Science 2012

Pusat Khidmat MakJumat Akademik ALAYSIA SA Atlt

ECTOPARASTE FAUNA OF RODENTS AND SCANDENTS IN DIFFERENT ECOLOGICAL HABITATS AND ITS POTENTIAL

ZOONOTIC IMPLICATIONS IN MALAYSIA

MADINAH BINTI ADRUS

A thesis submitted in fulfillment of the requirement for the Degree of

Master of Science

Faculty of Resource Science and Technology UNIVERSITI MALAYSIA SARAWAK

2012

DECLARATION

I hereby declare that all the work presented in the thesis is entirely my own work and has not

been submitted in support of an application for another degree or qualification to this or any

other university or institute of higher learning

(Madinah Binti Adrus)

Date

ii

DEDICATION

I dedicate my work to my beloved parents family mentors and all my loving

friends

111

ACKNOWLEDGEMENT

Alhamdullilah and thank Allah SWT for the bless and grace given to me for completing this

thesis successfully First of all I would like to thank my principle supervisor Professor Dr

Fatimah Abang and my co-supervisors Dr Mariana Ahamad and Professor Dr Mohd

Tajuddin Abdullah for their guidance encouragement advice concern support and

constructive comments during this study

I would like to thank the Faculty of Resource Science and Technology Universiti Malaysia

Sarawak (UNIMAS) and the Institute for Medical Research Kuala Lumpur (lMR) giving me

the various logistic and administrative supports during this study Without the collaboration

between Universiti Malaysia Sarawak (UNIMAS) and Institute for Medical Research Kuala

Lumpur (IMR) this study could not be completed successfully I really appreciated the

experience and knowledge gained during my training in acarology module diploma in

applied parasitology and entomology course and identification of acarine ectoparasites at

Institute Medical Research Kuala Lumpur (21 July -7 August 2009) All knowledge and

experience from the training were very useful to complete this study

I would like to thank UNIMAS for granting me the Zamalah scholarship 2009111 for my

postgraduate financial support This study was mostly supported from the research project

funded by a UNIMAS top-down grant on the zoonosis and EID [E14006 IF07 106 IZRC 103

12007 (03)] and FRGSOl(23)17661201O(47) led by Professor Dr Mohd Tajuddin Abdullah

and colleagues I would also like to thank Sarawak Forestry Corporation and Sarawak

Forestry Department for granting permission under the State Wild Life Protection Rules 1998

and permit number NCCD 9074(IV)29 to enter and collect sample in protected areas in

Sarawak use the faci tities and conduct research and warm hospitality during the field

IV

1

sampling My thanks also go to the Department of Wildlife and National Park (DWNP)

Kuala Lumpur Sabah Wildlife Department and Sabah Forest Department Kota Kinabalu

Sabah for their kind hospitality during my study in Peninsular Malaysia and Sabah

Within two years I have been fortunate to learn about the meaning of friendship I would like

to thank all members of the Department of Zoology Mr Wahap Marni Mr Mohd lalani

Mortada Mr Isa Sait Mr Huzal Irwan Husin Mr Trevor Allen and Mr Besar Keto for

their hard work and assistance throughout this study Besides that special thanks to Mrs

Halimaton Ibrahim Mr Mohd Kulaimi Bujang and all staff in the Acarology Unit of Institute

for Medical Research (lMR) for their guidance during the acarology training

To the Department of Zoology colleagues Muhamad Ikhwan Idris Kishen Bunya

Christharina Gintoron Irene Christianus Roberta Chaya Tawie Mohd Ridwan Abd

Rahman Pang Sing Tyan Sigit Wirantoro Nor Haliza Hasan Eileen ak Lit Mohd Fizl Sidq

Nur Salmizar Zahirunisa Abdul Rahim Debby Lim Siti Zuriani Ismail Wan Nurainie Wan

Ismail Farhana Anwarali Khan Badiozaman Sulaiman Mohd HanifRidzuan Mat Daud Nur

Aida Tamrin and Mr Faisal Anwarali Khan I wish to thank them for their wonderful

friendship for the great times shared knowledge and support

I would like to take this opportunity to thank those who are closest to me Firstly my loving

father Adrus Bakarudin and mother Rohana Hj 10lhi whose tender support and prayers and

to my brothers Fadillah Hussien and Abdul Ar-Razzaaq who always motivate me in

completing this thesis Therefore finally I dedicated this thesis for them

v

ABSTRACT

Field surveys of ectoparasites on Rodentia and Scandentia were conducted in 16 different

locations in Malaysia from June 2008 to May 20 I O The objectives of this study were to

determine the taxonomic composition of ectoparasites on Rodentia and Scandentia to

determine the relationship between ecological diversity and the host-parasite in different

types of ecological habitat in Malaysia and to examine whether the presence of ectoparasites

(ticks) have any potential health risks of rickettsial disease to local communities living

around the study area A total of 142 small mammals comprising of four species of squirrels

(Sciuridae) 12 species of rats (Muridae) and seven species of treeshrews (Tupaiidae) were

captured and examined for ectoparasites The hosts were derived from two orders namely

Rodentia (Sciuridae and Muridae) and Scandentia (Tupaiidae) The majority of these captures

was predominated by the Mullers rat (Sundamys muelleri) followed by Brown spiny rat

(Maxomys rajah) and Common treeshrew (Tupaia glis) Of these hosts six groups of

ectoparasites were recovered in this study namely ticks mesostigmatid listrophorid

chiggers lice and itch mites Two orders (Acarina and Anoplura) and seven families of

ectoparasites were present in this study namely Ixodidae Laelaptidae Trombiculidae

Atopomelidae Sarcoptidae Hoplopleuridae and Polyplacidae Twenty-three genera with 47

species of ectoparasites of Rodentia and Scandentia were recorded The ectoparasites were

sampled from all the seven types of habitats namely secondary forest lowland dipterocarp

forest riverine forest peat swamp mixed dipterocarp forest hill dipterocarp forest and

limestone forest Among the 16 localities secondary forest recorded the highest number of

host caught ectoparasites collected and with high infestation while the limestone forest was

found to have the highest number of host species and ectoparasite species compared to the

other five habitats Among the ectoparasites found Dermacentor sp Haemaphysalis sp

VI

I

Ixodes sp 1 granulatus Laelaps nuttalli Leptotrombidium sp Leptotrombidium deliense

Sarcoptes scabiei Polypax spinulosa and Hoplopleura dissicula are of known medical

importance Fifty individuals of ticks collected from the 16 locations of study sites (44

individuals) and museum collections (six individuals) were used for detection of Rickettsia

species The results of Polymerase Chain Reaction (PCR) detections were negative showing

that there was no infection of ticks by Rickettsia species

Keywords ectoparasites Rodentia Scandentia host ecological habitat Polymerase Chain

Reaction (PCR)

Vll

Ektoparasit Fauna ke atas Roden dan Scanden pada Ecologi Hahitat yang Berlainan dan

Pemhahitan dalam Potensi Zoonotik di Malaysia

Ahstrak

Tinjauan ke atas ektoparasit pada Rodentia dan Scandentia telah dijalankan di J6 kawasan

berainan di Malaysia dari bulan Jun 2008 sehingga bulan Mei 2010 Tujuan kajian ini

dijalankan adalah untuk mengenalpasti komposisi taksonomi ke atas ektoparasit pada

Rodentia dan Scandentia untuk mengenalpasti hubungan di antara kepelbagaian ekolog dan

perumah-parasit di kawasan berlainan ekologi habitat di Malaysia dan untuk mengllji sama

ada kehadiran ektoparasit (sengkenit) berkemungkinan untuk menyebar penyakit yang

disebabkan oleh Rickettsia dan seterusnya berpotensi untuk membahayakan kesihatan

penduduk yang tinggal di kawasan kajian Sejumlah J42 mammalia kecil yang terdiri

daripada empat spesies tupai (Sciuridae) J2 spesies tikus (Muridae) dan tlljuh spesies tupai

muncung (Tupaiidae) telah ditangkap dan diperiksa untuk kehadiran ektoparasit Kesemua

perumah telah dikenalpasti tergolong dalam dua order iaitu Rodentia (Sciuridae dan

Muridae) dan Scandentia (Tupaiidae) Hasil tangkapan ini adalah didominasikan oleh tikus

Muller (Sundamys muelleri) diikuti oleh tikus cokelat berbulu duri (Maxomys rajah) dan

tupai muncong besar (Jupaia glis) Hasil daripada perumah yang di tangkap ektoparasit

yang dikenalpasti dalam kajian ini terdiri daripada enam kumpulan iaitu sengkenit hama

mesostigmatid listrophorid tungau kutu dan hama gatal Dua order (Acarina dan

Anopura) dan tujuh famili ektoparasit telah dikenalpasti dalam kajian ini iailu Ixodidae

Laelaptidae Trombiculidae Atopomelidae Sarcoptidae Hoplopleuridae dan Polyplacidae

Dua puluh tiga genus dengan 47 spesies ektoparasit pada Rodentia dan Scandentia telah

direkodkan Ektoparasit telah diperolehi daripada kesemua tujuh jenis habitat iaitu hutan

viii

sekunder hutan dipterokarp tanah pamah hutan sungai paya gambut hutan dipterokarp

campuran hutan dipterokarp bukit dan hutan batu kapur Di antara 16 lokasi yang dikaji

hutan ekunder merekodkan hasil tangkapan yang tertinggi bagi perumah ektoparasit dan

kadar serangan manakala hutan batu kapur didapati mempunyai bilangan yang tertinggi

bagi spesies perumah dan spesies ektoparasit berbanding dengan lima habitat yang lain Di

antara ektoparasit yang dijumpai ialah Dermacentor sp Haemaphvsalis sp Ixodes sp l

granulatus Laelaps nuttalli Leptotrombidium sp Leptotrombidium deliense Sarcoptes

scabiei Polypax spinulosa dan Hoplopleura dissicula yang dikenalpasti mempunyai

kepentingan perubatan Lima puluh ekor sengkenit yang dikumpulkan dari 16 lokasi kajian

(44 individu) dan koleksi dari muzium (enam individu) telah digunakan untuk mengesan

kehadiran spesies Rickettsia Keputusan Tindakbalas Rantai Polimerase menunjukkan

keputusan negatif dan liada jangkitan spesies Rickettsia ke alas sengkenit yang diuji

Kata cunei ektoparasit Rodentia Scandenlia perumah ekologi habitat Tindakbalas Rantai

Polimerase

IX

TABLE OF CONTENTS

Title Page

Declaration II

Dedication 111

Acknowledgements IV

Abstract VI

Abstrak Vlll

Table ofContents x

List of Tables XVI

List of Figures XVII

List of Abbreviations XXI

Chapter 1 General Introduction

Research Overview

Objectives 3

Chapter 2 Literature Review

Ectoparasites 4

Category of Ectoparasi tes 5

Hosts ofEctoparasites 8

Previous Study of Ectoparasites 8

Ectoparasites as Carriers of Disease 12

Rickettsia species 14

x

26 The Effects of Rickettsia species 15

27 Previous Reports of Rickettsia in Malaysia 15

Chapter 3 General Materials and Methods

31 Study Sites 17

32 Host Collection 19

33 Processing Animals 21

34 Mounting ofEctoparasites 22

35 Identification ofEctoparasites 22

36 Detection of Rickettsiae in Selected Ticks 23

361 Isolation of DNA from Selected Tick Samples 23

362 Polymerase Chain Reaction (PCR) Amplification of Rickettsiae from 2S

Selected Tick Samples

Chapter 4 Taxonomic Composition of Ectoparasites on Rodentia and

Scandentia in Malaysia

Introduction 28

Materials and Methods 29

Results 313

43 t Taxonomic Diversity 31

432 Geographical Distribution 32

433 Systematic Accounts 34

4331 Key to the Orders of Ectoparasites Collected in this Study 34

xi

4332 Families of Ectoparasites Collected in this Study 34

43321 Key to Families of the Order Acarina (Ectoparasitic Acarine) 34

43322 Key to Families of the Order Anoplura (Ectoparasitic Insect) 37

4333 Genera of the Ectoparasites Collected in this Study 37

43331 Key to Genera of the Family Ixodidae 37

43332 Key to Genera of the Family Laelapidae 49

433321 Key to the Species ofLaeaps in Malaysia 57

43333 Key to Genera of the Family Trombiculidae 85

43334 Genera of the Family Atopomelidae 100

43335 Genera ofthe Family Sarcoptidae 113

43336 Key to Genera ofthe Family Hoplopleuridae 117

43337 Key to Genera of the Family Polyplacidae 119

Discussion 126

Conclusion 130

Chapter 5 Ecological diversity and host-parasite relationships in Rodentia and

Scandentia

Introduction 131

Materials and Methods 133

521 Description of Study Area 133

5211 Bukit Aup Jubilee Park Sibu Sarawak 134

5212 Sungai Menyarin Lanjak Entimau Wildlife Sanctuary Sarawak 134

5213 Sungai Bloh Lanjak Entimau Wildlife Sanctuary Sarawak 136

XII

5214 Kampung Giam Padawan Sarawak 136

5215 Endau Kluang Wildlife Reserve lohore 136

521 6 Kubah Nationat Park Kuching Sarawak 137

5217 Balambangan Island Sabah 137

5218 Tasek Bera Pahang 138

5219 Rumah Temuai Nanga Merit Kapit Sarawak 138

521 10 Sungai Beletik Nanga Merit Kapit Sarawak 139

52111 Gunung Regu Padawan Sarawak 139

52112 Sungai Dusun Wildlife Reserve Selangor 139

52113 Lata Bujang Krau Wildlife Reserve Pahang 140

521 14 Bukit Pueh Sematan Sarawak 140

521 15 Human settlement Kanowit Sarawak 141

52 116 Niah National Park Miri Sarawak 141

522 Methodology 141

Results 142

531 Taxonomic Composition of Rodents and Scandents 142

532 Infestation Rates in Small Mammals 144

533 Taxonomic Composition of Ectoparasites Collected from Host 144

5331 Ticks 144

5332 Mesostigmatid Mites 146

5333 Chiggers or Larval of Trombiculid Mites 149

5334 Lice 149

5335 Listrophorid Mites 150

X111

5336 Astigmatid mites (Itch Mites) 150

534 The Relationships of Hosts and Ectoparasites in Different Ecological Habitats 151

5341 The Host in Different Habitats 151

5342 Ectoparasites in Different Habitats 153

53421 Secondary Forest 153

53422 Limestone Forest 153

53423 Hill Dipterocarp Forest 155

53424 Lowland Dipterocarp Forest 158

53425 Mixed Dipterocarp Forest 158

53426 Peat Swamp 158

53427 Riverine Forest 159

Discussion 159

Conclusion 167

Chapter 6 Detection of Rickettsia Species in Ectoparasite (Tick) Samples

Introduction 168

Materials and Methods 170

621 Tick Samples Used for Test 170

622 Detection ofRickettsia Species in Selected Ticks 173

173Results

Discussion 175

Conclusion 177

xiv

Chapter 7 General Discussion and Conclusion

General Discussion 178

711 Ectoparasite Composition 178

712 Host-parasite Relationships in Contrasting Ecological Habitats 180

713 Potential Zoonotic Ectoparasites 181

Conclusion 182

Future Direction and Recommendation 183

REFERENCES 185

APPENDIX 205

xv

LIST OF TABLES

Table 31 Summary of 16 locations of study sites in Malaysia with coordinate and 19

habitat types of each location

16 study sites

genes

Table 32 Summary of dates of sampling total days sampling and methods used in 20

Table 33 Primers and nucleotide sequence used for amplification of rickettsial 25

Table 34 Component of mastermix and volume used for each sample 26

Table 35 The parameter of peR process for a total of 40 cycles 27

Table 41 Taxonomic list and relative abundance of ectoparasites collected from 16 33

study sites

rodents and scandents were collected from 2008 to 20 I O

Table 51 Locality information for 16 sites in Malaysia at which ectoparasites of 135

52 Taxonomic list of hosts and the number of parasite infestation in 16 sites 143

53 Ectoparasite infestation rates on rodents and scan dents in 16 locations in 145

Malaysia (2008-2010)

54 Taxonomic list and number of ectoparasites fauna found on rodents and 147

scandents in 16 sites (2008-2010)

55 Ecological habitat versus total number of rodents and scandents caught 152

56 Percentage of ectoparasite infestation in seven habitats 154

57 Total number of ectoparasites extracted from rodents and scandents In 156

seven different types of habitats

61 List of species field code host species localities and habitats of ticks 171

used in the detection of Rickettsia species

xvi

LIST OF FIGURES

Figure 31 Map of sixteen localities of sampling sites in Malaysia 18

Figure 41 Order Acarina (a) hypostome present in the mouthparts (b) spiracular plates present (ventral view of Ixodidae) (c) stigmata and peritremes present (ventral view of Laelaptidae) (d) scutum present (dorsal view of Trombiculidae) (e) tarsal suckers present in Sarcoptidae (ventral view) and (f) long body shape of Atopomelidae

36

Figure 42 Order Anoplura (a) stout spines present and (b) stout spines absent 37

Figure 43 Family Ixodidae (a) anal groove contouring the anus posteriorly (ventral view of genus Amblyomma) (b) anal groove contouring the anus anteriorly (ventral view of genus Ixodes) (c) palpi laterally beyond the basis caputuli (dorsal view of genus Haemaphysalis) (d) palpi long and slender (dorsal view of genus Amblyomma) and (e) basis caputuli hexagonal (dorsal view of genus Rhipicephalus)

39

Figure 44 Distribution ofAmblyomma spp in Malaysia 40

Figure 45 Distribution ofDermacentor spp in Malaysia 42

Figure 46 Distribution ofHaemaphysalis spp in Malaysia 44

Figure 47 Distribution of Ixodes spp in Malaysia 46

Figure 48 Distribution of Ixodes granulatus in Malaysia 48

Figure 49 Distribution ofRhipicephalus spp in Malaysia 49

Figure 410 Family Ixodidae (ticks) (a) Amblyomma sp ~ (dorsal view) (b) Dermacentor sp ~ (ventral view) (c) Haemaphysalis sp ~ (ventral view) (d) Ixodes sp ~ (dorsal view) Ixodes granulatus ~ (ventral view) 0 (dorsal view) (f) Rhipicephalus sp ~ (dorsal view)

50

Figure 411 Family Laelapidae (ventral view) (a) genital shield with a pair of setae (b) genital shield with 3 or 4 pair of setae (c) hooklike structure on coxa II (d) no hooklike structure on coxa II (e) leg 1 not longer than leg II (f) leg 1 longer than leg II (g) transverse striated presternal area present and (h) transverse striated presternal area absent

52

Figure 412 Distribution ofEchinonyssus nasutus in Malaysia 53

Figure 413 Distribution ofHaemolaelaps audyi in Malaysia 54

Figure 414 Distribution ofHaemolaelaps traubi in Malaysia 56

XVll

Figure 415 Distribution of Haemolaelaps spp in Malaysia 57

(dorsal view) (b) Haemolaelaps audyi ~ (ventral view) (c) Haemolaelaps traubi ~ (ventral view) (d) Laelaps aingworthae ~

(ventral view) (e) Laelaps echidninus ~ (ventral view) (f) Laelaps flagellifer ~ (ventral view) (g) Laelaps insignis ~ (ventral view) and

Figure 416 Distribution ofLaelaps aingworthae in Malaysia 61

Figure 417 Distribution ofLaelaps echidninus in Malaysia 62

Figure 418 Distribution ofLaelapsflagellifer in Malaysia 64

Figure 419 Distribution ofLaelaps insignis in Malaysia 66

Figure 420 Distribution ofLaelaps mercedeae in Malaysia 68

Figure 421 Distribution of Laelaps nuttalli in Malaysia 69

Figure 422 Distribution ofLaelaps rex in Malaysia 70

Figure 423 Distribution ofLaelaps sanguisugus in Malaysia 73

Figure 424 Distribution ofLaelaps sculpturatus in Malaysia 75

Figure 425 Distribution ofLaelaps sedlaceki in Malaysia 79

Figure 426 Distribution of Laelaps turkestanicus in Malaysia 81

Figure 427 Distribution ofLongolaelaps longulus in Malaysia 83

FJ8UIe 428 Distribution ofLongolaelaps whartoni in Malaysia 84

Figure 429 Distribution ofHirstionyssus sp in Malaysia 85

Figure 430 Family Laelapidae (mesostigmatid mites) (a) Echinonyssus nasutus ~ 86

(b) Laelaps mercedeae ~ (ventral view)

figure 431 Family Laelapidae (mesostigmatid mites) (a) Laelaps nuttalli ~ 87 (ventral view) (b) Laelaps rex ~ (ventral view) (c) Laelaps sanguisugus ~ (ventral view) (d) Laelaps sculpturatus ~ (ventral view) (e) Laelaps sedlaceki ~ (ventral view) (f) Laelaps turkestanicus ~ (ventral view) (g) Longolaelaps longulus ~ (ventral view) and (h) Longolaelaps whartoni ~ (ventral view)

XVIll

Figure 432 Family Trombiculidae (a) anterior median seta present (b) anterior 89 median seta absent (c) scutum with anterolateral seta shoulders (d) posterolateral setae short than anterolateral setae and (e) anterolateral setae short than posterolateral setae

view) (b) Gahrliepia (gahrliepia) sp (dorsal view) (c) Gahrliepia (walchia) sp (dorsal view) (d) Leptotrombidium deliense (dorsal view (e) Leptotrombidium sp (dorsal view) (t) Siseca sp (dorsal view) (g) Walchiella sp (dorsal view)

Figure 433 Distribution of Ascoschoengastia spp in Malaysia 91

Figure 434 Distribution of Gahrliepia (gahrliepia) spp in Malaysia 92

Figure 435 Distribution of Gahrliepia (walchia) spp in Malaysia 93

Figure 436 Distribution of Leptotrombidium deliense in Malaysia 95

Figure 437 Distribution ofLeptotrombidium spp in Malaysia 96

Figure 438 Distribution ofMicrotrombicula spp in Malaysia 97

Figure 439 Distribution of Siseca spp in Malaysia 99

FIgUre 440 Distribution of Walchiella spp in Malaysia 100

Figure 441 Family Trombiculidae (chiggers) (a) Ascoschoengastia sp (dorsal 10 I

figure 442 Trochanter without setae (genus L istroph oro ides ) 102

Distribution ofListrophoroides sp1 in Malaysia 104

Distribution ofListrophoroides sp2 in Malaysia 106

Distribution ofListrophoroides sp3 in Malaysia 107

Distribution ofListrophoroides spA in Malaysia 108

Distribution ofListrophoroides sp5 in Malaysia 109

Distribution ofListrophoroides sp6 in Malaysia 110

Distribution ofListrophoroides sp7 in Malaysia III

Distribution ofListrophoroides sp8 in Malaysia 112

Distribution ofListrophorid 1 (unidentified) in Malaysia 113

xix

Figure 452 Family Atopomelidae (listrophorid mites) (a) Listrophoroides spt (b) 114 Listrophoroides sp2 (c) Listrophoroides sp3 (d) Listrophoroides sp4 (e) Listrophoroides sp5 (t) Listrophoroides sp6 (g) Listrophoroides sp7 (h) Listrophoroides sp8 (i) Listrophorid 1 (Unknown)

Figure 453 Tarsal sucker present (genus Sarcoptes) 115

Figure 454 Distribution ofSarcoptes scabiei in Malaysia 116

Figure 455 Sarcoptes scabiei (itch mites) (ventral view) 116

Figure 456 Family Hoplopleuridae (a) antenna four segmented and (b) antenna 117 five segmented

Figure 457 Distribution ofAncistroplax nasuta in Malaysia 118

Figure 458 Distribution ofHoplopleura dissicula in Malaysia 120

Pipre 459 Family Hoplopleuridae (a) Ancistroplax nasuta (ventral view) and (b) 120 Hoplopleura dissicula (ventral view)

Figure 460 Family Polyplacidae (a) tubercles present and (b) tubercles absent 121

Figure 461 Distribution ofNeohaematopinus callosciuri in Malaysia 122

Distribution of Polypax spinulosa in Malaysia 123

Distribution ofSathrax sp in Malaysia 124

Family Polyplacidae (a) Neohaematopinus callosciuri (ventral view) 125 (c) Polypax spinulosa (ventral view) (d) Sathrax sp (dorsal view)

Map of Malaysia and the 16 localities where ectoparasites of rodents 133 and scandents were sampled during the present study

Maps showing the 16 localities of selected ticks used in the detection 170 of rickettsial disease in Malaysia

Polymerase chain reaction products 174

xx

ABBREVIATIONS

bp base pair

BAJP Bukit Aup Jubilee Park

BIS Balambangan Island Sabah

BPS Bukit Pueh Sematan

degC degree Celcius (temperature)

DNA deoxyribonucleic acid

dNTP deoxyribonucleotide triphosphate

ddH20 double-distilled water

EtBr ethidium bromide

EKWR Endau Kluang Wildlife Reserve

g gram

GRP Gunung Regu Padawan

HCI hydrochloric acid

HSK Human Settlement Kanowit

KGP Kampung Giam Padawan

km kilometer

km2 kilometre square

KNP Kubah National Park

LBKWR Lata Bujang Krau Wildlife Reserve

M molar

MgCh magnesium chloride

ml mililitre

XXI

mm

mM

mtDNA

mg

nm

NP

NR

NNP

peR

PBS

RNase

rpm

RTNM

SBNM

SBLEWS

SDWR

SMLEWS

TAE

TBE

TE

TBRS

Jil

metre square

millimetre

millimolar

mitochondrial deoxyribonucleic acid

milligram

nanometre

National Park

Nature Reserve

Niah National Park

polymerase chain reaction

phosphate buffered saline

ribonuclease

rotation per minute

Rumah Temuai Nanga Merit

Sungai Beletik Nanga Merit

Sungai Blob Lanjak Entimau Wildlife Sanctuary

Sungai Dusun Wildlife Reserve

Sungai Menyarin Lanjak Entimau Wildlife Sanctuary

tris-acetate-etbylenediaminetetraacetic

tris borate EDT A

tris EDTA

T asek Bera Ramsar Site

microlitre

xxu

flM micromolar

UNIMAS Universiti Ma1aysia Sarawak

UV ultraviolet

V volt

xxiii