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Opportunistic Sampling as a Tool for Surveillance of Parasitic Diseases in Wild Animals; a Case of Mikumi Natonal Park

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SOKOINE UNIVERSITY OF AGRICULTURE

FACULTY OF FORESTRY AND NATURE CONSERVATION DEPARTMENT OF WILDLIFE MANAGEMENT SPECIAL PROJECT PROPOSAL TITLE: OPPORTUNISTIC SAMPLING AS A TOOL FOR SURVEILLANCE OF PARASITIC DISEASES IN WILD ANIMALS; A CASE OF MIKUMI NATONAL PARK

STUDENT NAME: KIBONA, Juma A REG. NUMBER: WLM/D/11/T/0057 A SPECIAL PROJECT PROPOSAL SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE BARCHELOR OF SCIENCE IN WILDLIFE MANAGEMENT FROM THE SOKOINE UNIVERSITY OF AGRICULTURE, MOROGORO – TANZANIA

SUPERVISOR: Prof. E.K. BATAMUZI 2013/2014

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Table of Contents
1.0 INTRODUCTION................................................................................................................................. 2 1.1 BACKGROUND OF INFORMATION ............................................................................................. 2 1.2 Statement of the Research Problem .................................................................................................... 4 1.3 OBJECTIVE ....................................................................................................................................... 6 1.3.0 General objective ............................................................................................................................. 6 1.3.1 Specific objectives ....................................................................................................................... 6 1.4 RESEARCH QUESTIONS................................................................................................................. 6 2.0 LITERATURE REVIEW ................................................................................................................... 7 2.1 General overview of diseases .............................................................................................................. 7 3.0 RESEARCH METHODOLOGY ........................................................................................................ 8 3.1 Description of study area .................................................................................................................... 8 3.1.1 Location of study ......................................................................................................................... 8 3.1.2 Wild animals .............................................................................................................................. 10 3.1.3 Climate ....................................................................................................................................... 10 3.1.4 Landforms, topography, and biodiversity .................................................................................. 10 3.2 Materials to be used .......................................................................................................................... 11 3.3 Study design ...................................................................................................................................... 11 3.4 Sample collection .............................................................................................................................. 12 3.4.1 Ectoparasites .............................................................................................................................. 12 3.4.2 Faecal samples ........................................................................................................................... 12 3.4.3 Biopsy samples .......................................................................................................................... 12 3.4.4 Laboratory assessment of the samples ....................................................................................... 13 4.0 DATA ANALYSIS AND PRESENTATION .................................................................................... 13 4.1 Statistical Data analysis .................................................................................................................... 13 4.2 Data Presentation .............................................................................................................................. 13 7.0 REFERENCES .................................................................................................................................... 14 8.0 List of appendices................................................................................................................................ 18

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1.0 INTRODUCTION 1.1 BACKGROUND OF INFORMATION Parasitic diseases are infectious diseases caused or transmitted by parasites, but many parasites do not cause disease. Practically all living organisms can be affected by parasitic diseases including plants and animals but this study is much interested in animals (wildlife). Disease surveillance is an important tool in disease detection programmes to investigate which diseases present in wildlife settings and the further assessment of risks involved as well as the prevalence of disease and patterns of new and emerging conditions as an early warning system for diseases in wildlife (Worthington and Bigalke, 2001) The surveillance of parasitic disease in the wildlife setting is an important means for immediate recognition and research to characterize the nature of the host-pathogen interaction and to reexamine the potential risk of spread out or cross over infections to domestic animals populations since (Morner et al, 2002). Wildlife are generally considered to transmit diseases to human and other domestic animals that is why it is important for this study to carry out parasitic diseases detection through opportunity from the highway in order to know the health status of our wild animals Wildlife diseases are of growing concern worldwide because they threaten not only wildlife populations, but also domestic animals and human health (Vallet, 2008; Wentworth, 2008) In review of this, surveillance of parasitic disease is of great important as early warning and a means for planning control or eradication programmes. With thinning natural habitats and increasing numbers of threatened species worldwide, the surveillance wildlife diseases has become an essential component in the management of free-ranging wildlife (Morner et al, 2002) Wildlife may provide a reservoir of parasitic infections for some diseases but may also act as a sensitive indicator of diseases that are not clinically perceptible in adjacent livestock populations. In detecting the presence of a disease, characterizing its prevalence, spread and monitoring its progression in the wildlife population disease surveillance and monitoring in wildlife population always involved (Woodford, 2002)
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In sampling wild animals for disease surveillance different techniques are been used, Invasive and non-invasive. In fact the invasive method is of many disadvantageous since always it disturb animals, involves handling and manipulation of the animal, Example darting/immobilization of animals for the purpose of taking samples. The technique can cause death to animal, exposes animal to infections and needs a lot of money to run the exercise Opportunistic sampling as a non-invasive technique is an important tool for monitoring disease in wildlife populations. This involves the researcher taking advantage of unexpected opportunities in the course of the fieldwork, using available encounters and events as they arise. This needs clear knowledge on which sample belong to which animal because some time it involve taking of samples in absence of animal concern. Various opportunities can be used by researcher in taking biological samples from animal for diseases investigation purposes, such as natural death, Infrastructures such as road kills and opportunity for live animal sampling which arises when wildlife are captured for relocation and restocking efforts (Jessup et al. 1993) Like humans and other animals, wild animals suffer from diseases and parasites which can be lethal wildlife and to other domesticated animals including humans since wildlife are known to be a potential source of human infections such Tuberculosis and monkey pox, especially to infants and young children, the elderly, and those with weakened immune systems (Wikipedia, the free encyclopedia.htm). It is typically difficult to vaccinate wild animals against these diseases because vaccines developed for domestic animals have not been proved to be effective on wild animals Wild animals also harbor parasites such as ascaris worms, tape worms, flukes, and protozoa that can cause debilitating and often fatal diseases in humans, while external parasites such as ticks and fleas vector-borne diseases in cats and dogs and other serious diseases. Highways in mikumi national park reported have serious impacts to the wildlife and their environment due to some pollutants from highways being introduced to wildlife settings In wild animals to obtain samples is a challenging task due to difficulties and handling the animal, Cost of immobilization, risk of animal attack and scavengers compared to domestic animals. Due to lack of information on how highways can be used as an opportunity to collect

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biological samples this study aim to use highway as an opportunity to collect samples from animals for parasitic disease detection 1.2 Statement of the Research Problem Wild animals like other animals are affected by parasites .The parasite affecting different animal species are resulting from the ecological interaction of wild animals and the environment favorable in hosting parasites which affect the health status of wild animals. Due to increased rate of environmental pollution in wildlife ecosystem the animals occupying such ecosystem become infested with a number of diseases causing pathogens including the intestinal worms Parasites are highly diverse group of organisms that have evolved different strategies for infecting their host. Some parasites such as lice and ticks are found on external part of the animal’s body (ectoparasites) but most of the parasites are found internally (endoparasite).Some parasites can be seen by naked eyes (macroscopic) while others are microscopic, such as protozoans that cause avian malaria. Many parasite species have been detected from free-ranging wild birds (Spalding, 1993); however the presence of parasite does not necessarily associated with diseases due to the fact that parasites in wildlife in protected areas considered being part of a natural functioning ecosystem Parasitic diseases are of economic importance in a sense that they affect the productivity of animal through deaths, by decreasing growth rate, efficiency in feed conversion ratio and levels of the total plasma proteins due to a fall in absorbed amino acids that are essential for protein synthesis as well as lowering the quality of the meat (Alexandre and Mandonnet , 2005) .In ensuring our wildlife remain healthy and productive disease detection is of great importance for the benefits of our nation

Surveillance of parasitic disease in wild animals is a challenging task; this is due to the fact that the infected animals do not display the signals of being sick. Sometimes the animals seems to be physically fit and run away and some time the animal tend to hide when is sick. A number of significant challenges also arise in detecting some parasitic disease in a free-ranging wildlife, even in susceptible species; the majority of infected animals show no clinical signs of disease.

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It is probably adaptive for wild animals not to disclose when they are hurt or sick because predators and competitors often attack the sick and the weak animal

According to Morner et al, (2002), it is more difficult to detect diseases in wildlife than in domestic animals because wild animals are not constrained by boundaries and can roam over large distances so to handle the animal in order to obtain samples becomes a challenging task. The scavengers is also a challeng,when the animal die scavengers such as hyenas and vultures eat the animal which make collection of samples difficult, the whole carcass may be eaten

Statistic show that 2-4 animals are killed daily due to road accident in the park.Research conducted on collection of vital samples from an immobilized giraffe as part of an ear and skin disease investigation in Mikumi National Park on ecosystem health. (Kazwala et al, 2009). The disease was first observed in 1999 in two giraffes but now has spread to affect giraffes throughout Mikumi NP and is spreading further south into Selous GR, the largest wildlife area in Africa (Osofsky, 2005) Construction of highways contributes to economic development through improvement of living conditions and expansion of social overhead capital. But on other hand, they influenced the fragmentation and deterioration of wildlife habitat (Bennett et al, 1994; Trombulak and Frissell, 2000). In Mikumi National Park some researches and investigations have been done on the impact of highways to wildlife s and diseases which affect the health of the animals and productivity by lowering the immune system of the animal but collecting samples from road kills and investigate for the parasitic diseases not yet done

The 50Km road and 50m away from the road in both sides will provide an opportunity to sample the animals for parasitic diseases surveillance through the road kills so that to provide a warning for diseases in wild animals in mikumi ecosystem. Early intervention and investigation of unusual or unexpected disease events are essential to the goal of determining the cause and significance of the disease outbreaks in the protected area. Report of this nature may be the first indication of the introduction of an exotic disease agent in wildlife settings

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This study aim at identifying the disease causing agents (Parasites) from sampled road kills in mikumi ecosystem, the disease that affect the health of the animals as the result of environment stress especially due to human interaction with wild environment through the road and other highways which pass through the park The implication of this study is yielding information that will be useful in conservation, disease surveillance and monitoring of wild animals health status in Mikumi National Park and Tanzania in general Information on the occurrence of parasitic disease in wildlife is essential to both the local and global understanding of free-living animal population (Artois et al, 2001). Report of illness or deaths involving many animals from a free-living population may represent the initial alert to the likely presence of a new disease agent (Woodford, 1982) 1.3 OBJECTIVE 1.3.0 General objective The overall objective is to apply opportunistic sampling in parasitic disease surveillance and characterization and parasitic disease in wild animals from road kills in Mikumi national park. 1.3.1 Specific objectives 1. To carry out opportunistic sampling for parasitic disease detection along the highway within a Park 2. To identify and characterize disease causing agents from sampled road kills 1.4 RESEARCH QUESTIONS 1. Can a 50 km road provide opportunity to sample the wild animals from road kills? 2. What samples are to be collected for laboratory investigation? 3. What are the parasites in Mikumi National Park? 4. Is there any disease condition in wild animal’s samples from road kills in Mikumi National Park?

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2.0 LITERATURE REVIEW 2.1 General overview of diseases Many of the emerging diseases around the world affect large ecosystems and cross country boundaries, including those of Tanzania, there is a need to strengthen local capability to detect and identify disease threats in protected areas, to launch efficient reporting mechanisms of the disease, and to develop intensive efforts to manage and mitigate their effects (Keusch et al, 2009). Tanzania National Parks (TANAPA) has been developing a wildlife veterinary unit to address these numerous and emerging wildlife health challenges in the country so as to ensure that the wild life remain healthy and productive. According to the World Organization for Animal Health (OIE), Terrestrial Animal Health Code (2006), Disease surveillance in animal health is an essential component necessary to detect diseases, to monitor disease trends, to control endemic and exotic diseases, and to support claims of freedom from disease (OIE, 2006). According to Karesh (2008), wild animals are more susceptible to new parasitic diseases than domesticated animals due to the way their natural ecosystem functioning which make easy to detect the presence of disease and are good indicators of emerging disease outbreaks in the protected areas. The clinical signs of disease in wildlife are not easily observed as in domestic animals, and the hands-on collection of samples from specimens is more difficult. These factors make surveillance and response to disease outbreaks sometimes much difficult to implement in wildlife disease surveillance (Vallat, 2008). Highways also directly reduce wildlife populations through mortality and habitat loss (Fahrig et al, 1995 and Forman, 2000). Habitat loss is further increased for species that avoid habitats near roads (Reijnen et al, 1996). In Brazil, the majority of studies on highway mortality of the fauna have been descriptive resulting merely in lists of species without any standardization of the

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methods used or a clear definition of sampling effort of wild animals for disease detection (Bager et al, 2007). The researches done in Mikumi National park indicates that wild animals species found in the park are infected by different parasitic diseases that affects the animal health, reducing growth rates or fecundity, increasing metabolic requirements, changing patterns of behaviour and some severe parasites infestation ultimately may cause death to the animal. Research conducted on collection of vital samples from an immobilized giraffe as part of an ear and skin disease investigation in Mikumi National Park on ecosystem health (Kazwala et al, 2009). Also environmental stress, habitat fragmentation, climatic changes which mainly on temperature changes, predators’ pressure, and competition with other animals occupying the same habitat and human activities impacts the wildlife in Mikumi National Park

3.0 RESEARCH METHODOLOGY 3.1 Description of study area 3.1.1 Location of study This study will be conducted at Mikumi National Park found in Morogoro region located along Latitude 60 51' South and Longitude 31 41' East at an altitude of 530 m above the sea level. The Samples that will be used in this study will be collected from wild animals in the park which is located 283 km (175 miles) west of Dar es Salaam, north of Selous, and in the route to Ruaha, and Udzungwa national parks

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Fig: 1 Map of Mikumi National Park Showing High Ways. (TANAPA) The park established in 1964, currently covers an area of 3230 km² (1,250 sq miles), the fourthlargest park in Tanzania, and part of a much larger ecosystem centered on the uniquely vast Selous Game Reserve and it lies between the villages of doma and mikumi on the Dare s salaam – Tunduma Highway, Mikumi national park falls into a gap in the eastern Arc mountains (http://www.tanzaniaparks.com/mikumi.html, Accessed on 2013/25 /10) Mikumi National Park is bisected by the Tanzania-Zambia highway as one of the major challenge of the park which crosses Mikumi National Park over a stretch of 50 km. The road was
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rehabilitated between May 1990-November 1991 resulting in higher average driving speeds of vehicles. High speed and the danger of collisions with wildlife are causing regular accidents in Mikumi National Park (DREWS, 1995) 3.1.2 Wild animals Wild animals expected to see in Mikumi National Park are, the African elephant (Loxodonta Africana), and the African hunting dog (Lycaon pictus), yellow baboons (Papio cynocephalus, Linnaeus), Maasai giraffe ( giraffe camelopadalis), and Reticulate giraffe ( giraffa reticulate), Blue wildebeest (Connochaetes taurinus),African buffalo (Syncerus caffer), Lion(Panthera leo),zebra (Equus quagga), impala (Aepyceros melampus), Hippopotamus (Hippopotamus amphibius), greater kudu (Tragelaphus strepsiceros), sable antelope (Hippotragus niger) and over 450 bird species. 3.1.3 Climate Morogoro region in general has both bimodal and unimodal rainfall patterns. The northern part has bimodal rainfall and, therefore, two growing seasons in a year. Short rains known in Swahili as vuli start between mid-September and mid-October and continue to December while long rains commonly known as masika start in mid-March to late May (Turner and Paavola, 2003) Unimodal rainfall dominates the southern part, which also includes Mikumi National Park. The rainfall pattern has one long rain season, which starts from December to May and a single dry season from June to November (Hawkins & Norton, 1998). March and April are the wettest months. 3.1.4 Landforms, topography, and biodiversity Mikumi national park falls into a gap in the eastern Arc Mountains which stretch from Kenya down to southern Tanzania and are seen as isolated ranges. The park is located in an area where four vegetation zones intersect making it a diverse ecotone. The four vegetation types are miombo woodland in the south, arid bush land in the north, coastal zone in the east and mountain climate in the east and west (Hawkins and Norton 1998). The miombo woodland consists of mainly Brachystegia spp, while Combretum-Terminalia woodland dominates between hill areas and in floodplain (Mercer, 1983). The park is also dominated by other species like Sclerocarya caffra, Cassia abbreviata, Borassus flabellifer and Hyphaene ventricosa palms. Balanites aegyptiaca and Ficus spp.The open grassland is a reliable
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site for Africa’s largest antelope, the skittish eland, while the woodland supports seldom-seen sable antelope and greater kudu. Predator sightings are less varied than in some parks in the Country, but lions are often seen lazing below shady acacias, spotted hyenas are vociferous by night, and endangered African wild dogs occasionally range across from the Selous. The pools situated north of the park headquarters where hippopotamus reside, usually supported by water birds 3.2 Materials to be used  Examination gloves for collecting biological samples         Alcohol 70% for preservation of parasites Formalin for preserving samples Transport medium Plastics bags for specimens’ collection GPS for taking the cardinal points Data sheet and notebook Field guide books for identification of parasites Digital Camera for taking photographs for reference

3.3 Study design A Cross sectional study whereby opportunistically data will be collected once Data will be collected opportunistically from the site namely Mikumi National Park along the 50Km stretches early in the morning during road patrol where samples from animals and samples from the environments will be collected. The choice of the site basically based on the fact that the Park is passed by the Dar es salaam-Tunduma road with high human activities including southands of vehicles using the road daily. The high speed of the moving vehicles collides with wild animals which eventually causes animal injury and death.

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3.4 Sample collection The samples will be collected using various materials and apparatus by the assistance of the staffs from road kill patrol group in Mikumi National Park. The samples from the dead or injured animals and environments are to be collected for Laboratory investigation of parasitic diseases in wild animals. 3.4.1 Ectoparasites The majority of these parasites will be collected just by picking them from the skin of infested animals. Examples, Lice, Fleas, Ticks some of mites which causes manges barrow inside the skin, to collect them scalpel blade to scrape them to get skin scraping specimens will be used. Ectoparasites will be stored in 70% Ethanol (ethyl alcohol).Mites can be stored in a piece of paper 3.4.2 Faecal samples Faecal samples will be collected directly from the rectum or from the ground (passive) by using examination gloves, and then turn the glove outside inside and tie up, or by using stool bottles. About 5grams of faecal material will be sufficient to perform the required tests. This is for diagnosis of gastrointestinal parasites such as worms, Giardia, Coccidia Sample will be Stored at +40C to +80C in a refrigerator, but not stored in a freezer, to prevent worm egg destruction. 3.4.3 Biopsy samples These are pieces of tissues or organs chopped from the animals. The biopsy can be a piece of liver, lymph node, skin or tissue mass. A clean cut of piece of tissue from animal body will be made and stored in whirl-Paks for fixed and fresh tissues and must be sealed correctly. The sample will be preserved in 10% formalin. In the field, transport medium and the cool box with cold ice packs will be used to transport the samples to Sokoine University Agriculture laboratory of Microbiology and Parasitology where identifications and characterization of the parasites will be done as a surveillance of diseases in animal body. A carefully observation using naked eyes and Microscope will be done.

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3.4.4 Laboratory assessment of the samples A qualitative laboratory sample analysis will be done where faecal samples will be processed and examined for Helminths and eggs identification. For faecal samples will be first screened for presence of Helminths eggs by use of wet smear preparation, 1-2 drops of normal saline will be put on glass slide and loopful of faecal sample added, mixed and examined at 10x eye piece on light microscope. Identification of eggs of different Helminths species will be done by using standard Helminths egg identification keys. Then the slide will be taken away and examined by using 10x eye piece and a 4x (40x total magnification) on a light microscope. Again, Helminths eggs will be identified based on their shape, size and color as shown standard Helminths egg identification keys. Tissue samples will be processed for laboratory analysis

4.0 DATA ANALYSIS AND PRESENTATION 4.1 Statistical Data analysis The data collected will be analyzed and arranged by using statistical methods which includes descriptive and quantification by tables and excel using simple statistics 4.2 Data Presentation The information to be collected from the study area will be presented in the specific day, time and place to be allocated by the responsible department at Sokoine University of Agriculture.

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7.0 REFERENCES Alexandre, G., & Mandonnet, N. (2005) Goat meat production in harsh environments Small Ruminant Research, 60(1), 53-66 Artois M., Delahay R., Guberti V. & Cheeseman C. (2001).-Control of infection diseases in wildlife in Europe. Vet.J. 162,141-145 Baskaran, N. & D. Boominathan (2010). Road kill of animals by highway traffic in the tropical forests of Mudumalai Tiger Reserve, southern India Journal of Threatened Taxa 2(3): 753-759. Bennet, A.F., K. Henein, and G. Merriam 1994 Corridor use and the elements of corridor quality: chipmunks and fencerows in a farmland mosaic. Biological Conservation 68, 155-165. Bennet, A.F., K. Henein, and G. Merriam 1994 Corridor use and the elements of corridor quality: chipmunks and fencerows in a farmland mosaic. Biological Conservation 68, 155-165. CHILDS, J.E. 2002. Epidemiology Pages 113-162 in A.C. Jackson and W.H. Wunner, editors Rabies Academic Press, San Diego, California Directions Ecological Economics 46(3): 493-510. Drews, C., 1995. Road kills of animals by public traffic in Mikumi National Park, Tanzania, with notes on baboon mortality. African Journal of Ecology 33, 89- 100 H. M.Munang’andu, V. Siamudaala, A.Nambota, M.Unyeme, and K. S. Nalubamba, “Detection of Babesis species in free-ranging puku, Kobus vordonii, on a game ranch in Zambia,”Korean Journal of Parasitology, vol. 166, no. 1-2, pp. 163–166,2009 Ornithology 64:169 – 178. Hawkins, D. and G. Norton (1998) Mikumi National Park: the unknown & neglected National Park, Animal Behaviour Research Unit (ABRU) Mikumi National Park, Tanzania.

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Hels, T. and Buckwald 2001 The effect of road-kills on amphibian populations Biological Conservation 99, 331-340 Jessup et al, 1993. Wildlife health implications in the translocation of wildlife In: Bissonette JA, Krausman PR eds. Integrating People and Wildlife for a Sustainable Future. Lawrence KS: Allen Press. P 381–385 Karesh, W. 2008 Africa needs warning systems for wildlife diseases. Keusch, G. T., Pappaioanou, M., Gonzalez, M. C., Scott, K. A., & Tsai, P. (Eds.) (2009) Sustaining global surveillance and response to emerging zoonotic diseases National Academies Press Kim, O. H. An, J. Song, and J. Lee 2001 The road kills damage of wild animal by fragmentation of habitat in Gyeongsangnam-do province, Dept. of Forest Resources, Chinju National University 14, 97-110. Knobel, d.l., s. Cleaveland, p.g. Coleman, e.m. Fever, m.l. Meltzer,m.e.g. Miranda, a. Shaw. J.zinsstag and f.x. Meslin 2005 reevaluating the burden of rabies in Africa and Asia Bulletin of the world health organization 83: 360-368 Laboratories Agency 2006 Diseases of Wildlife Scheme Morner, T., Obendorf, D.L., Artios, M., & Woodford, M.H. 2002 Revue Scientific Office International des Epizooties Surveillance and monitoring of wildlife diseases, 67-72 Noss R., (2004). The Ecological effects of Roads In PAW NET, 117 Main Street, Brattleboro, VT 05301 Olokesusi, F. (1990): Assessment of the Yankari Game Reserve, Nigeria: Problems and Prospects. Butterworth Heinemann Ltd., pp 153 – 155. Osofsky, S.A., Cleaveland, S., Karesh, W.B., Kock, M.D., Nyhus, P.J., Starr, L. and Yang, A. (Eds) (2005) Romin L. A. and Bissonette J. A. (1996) Temporal and spatial distribution of highway mortality of mule deer on newly constructed roads at Jordanelle Reservoir, Utah. The Great Basin Naturalist 56: 1-11.
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Spalding, M. G., & Forrester, D. J. (1993) Disease monitoring of free-ranging and released wildlife. Journal of Zoo and Wildlife Medicine, 271-280 Trumbo lak, S.C. and C.A. Fr issell 2000.Review of eco logical effects of roads on terrestrial and aquatic communities Conservation Bio logy 14, 18-30 Trumbulak, S.C. and C.A. Fr issell 2000 Review of eco logical effects of roads on terrestrial and aquatic communities. Conservation Bio logy 14, 18-30 Turner, R. K. and J. Paavola (2003) "Valuing nature: lessons learned and future research Vallat, B. 2008 OIE: Wildlife surveillance key to biosecurity. In: The Pig Site news. Veterinary Varland, D. E., Klaas, E. E., Loughin, T. M. (1993). Use of habitat and perches, causes of mortality and time until dispersal in post-fledging American Kestrels Journal of Field Williams, E.S., Yuill, T., Artois, M., Fischer, J., & Haigh, S.A. 2002 Emerging infectious diseases in wildlife In Infectious diseases of wildlife: detection, diagnosis and management (RG Bengis, Ed.). Revue Scientific Office International des Epizooties 21(1), 139 – 157 Woodford, M.H. 1982. Tuberculosis in wildlife in the Ruwenzori National Park, Uganda (Part II) Tropical Animal Health and Production, 14, 155-160 Worthington and Bigalke, 2001 R.W. Worthington and R.D. Bigalke, A review of the infectious diseases of African wild ruminants, Onderstepoor J. Vet Res. 68 (2001), pp.291–323 www. Wikipedia, the free encyclopedia.htm, accessed on 10th December 2013

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CERTIFICATE OF APPROVAL

Name of Student: KIBONA, Juma A Reg.Number: WLM/D/11/T/0057

Signature: ___________________ Date: ___________________

Name of Supervisor: ________________

Signature: ________________ Date: _____________________

Head of Department: ___________________ Signature: _____________________ Date: _________________________

Dean of the Faculty: __________________

Signature: _____________________ Date: _________________________

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8.0 List of appendices Appendex 1: Schedule of activities. Duration of the study is as shown below Activity/Month Oct 2013 Proposal writing Proposal presentation and submission Data collection Data analysis Report writing Report Submission Nov 2013 Dec 2013 Jan 2014 Feb. 2014 Mar 2014 Apr 2014 May 2014 Jun 2014

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Appendix 2: Break down of the projected budget for the study Source of funds to facilitate this work will be Higher Education Loan Board (HESLB) with the total loan amount of TShs. 180,000. The budget breakdown is shown below:

S/N 1 2 3 4

ITEMS Transport Accommodation and meal Stationery Research materials for data collection and analysis TOTAL

COST(TSHS) 15,000/= 80,000/= 35,000/= 50,000/= 180,000/=

Appendix 3 Sample of table shows the general information of sample to be collected S/N ORIGN SAMPLE

19…...

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