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Chapter One Introduction & Literature Review 1.1. General Introduction 1.2. Blood Parasites of Reptiles and Amphibians

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Chapter One Introduction & Literature Review 1.1. General Introduction 1.2. Blood Parasites of Reptiles and Amphibians Chapter one Introduction & literature review 1.1. General introduction The reptiles and amphibians can be infected by different type of parasites. These parasites can be divided into enteric parasites and ectoparasites. Enteric parasites include protozoa, flagellates, ciliates, opalinids, amoebae, and coccidea. Also they can be infected by bacteria from ectoparasites like mites and ticks (De la Navarre, 2009). 1.2. Blood parasites of reptiles and amphibians The blood parasites of the reptiles and amphibian may be divided into two classes depending on whether they infect the blood without attacking the corpuscles or whether they become established within the corpuscles; extracorpusular parasites such as the flagellates of the genera Trypanosoma, Leptomonas, and Leishmania, and nematodes, and intercorpuscular parasites such as many telosporidian species of the suborder Adeleiing belonging to the genera Karyolysus, Hepatozoon and Haemogregarina species (Reichenow, 1953). 1.3. Kinetoplastid protozoa This class contains species that parasitize a wide diversity of hosts ranging from humans to plants. Member of this group are characterized by a single large mitochondrion containing a body- the kinetoplast- that stain darkly in histological preparations. The Kinetoplastids differ considerably in their host distribution, life cycles, and medical and veterinary importance. (Battaglia et al. 1983) 1 1.3.1. Sauroleishmania spp Currently, the Leishmania parasites are classified into three subgenera, The subgenera Leishmania (Leishmania) which is transmitted by female sandfly of the Phlebotomus species in the Old World, and Lutzomyia species in the New World, the subgenera Leishmania (Viannia) which is only found in the New world and transmitted by Lutzomyia spp sanflies, and the subgenera Leishmania (Sauroleishmania) or Lizard’ leishmania, which is transmitted by the sandflies of Sergntomyia spp (Chappuis, et al., 2007). The Leishmania parasites have two forms in their life cycle, the promastigotes form in the sandfly, and the amastigotes form in the vertebrate host. In mammals, the organism is found in the macrophages and has a characteristic, distinctive, bar-shaped extra chromosomal DAN fragment called a kinetoplast. The motile promastigote form is found extracellularly and possesses a flagellum (Frye, 1991). The Sauroleishmania has been reported in the peripheral blood of reptiles, primarily in lizards (Keymer. 1981). The amastigote form in reptiles appears in the cytoplasm of blood cells, particularly the erythrocytes. It appears singly or may be numerous, and may condense to round basophilic inch with a central hollow (Paperna et al., 2001). Microscopic diagnosis is the standard and routine diagnosis for leishmaniasis, which depends on detection of Leishmanai amastigotes in Geimsa stained aspirate materials from lymph nodes, bone marrow, spleen or liver, and in slit skin smears or in peripheral blood, and/ or promastigotes in dissected materials of infected sandfly (Zijlstra, et al., 1992). 2 Hoare (1948) postulated that the lizard leishmania provide convincing evidence of evolution in which an invertebrate parasite has become adapted to life in a vertebrate host. 1.3.2. Trypanosoma spp These are large flagellate protozoa that possess a kinetoplast with a distinct or in distinct, undulating membrane (trypomastigote). These parasites may be found extracellularly in the peripheral blood of many reptiles’ species. They are transmitted by blood –sucking arthropods (biting dipteran flies such as Phlebtomine sandflies) in terrestrial reptiles, and by leeches in aquatic reptiles. Although trypanosomiasis can cause severe parasitemia, it is commonly associated with lifelong subclinical infection (Lane and Mader, 1996). All species of trypanosomatidae have a single nucleus and are either elongated with a single flagellum or rounded with a very short, no protruding flagellum. Many members of the family are heteroxenous: During one stage of their lives they live in the blood and/or fixed tissues of all vertebrate classes, and during other stages they live in the intestine of blood sucking invertebrates (Tibayrenc & Ayala. 1999). 1.4. Apicomplexa spp Apicomplexa species is a large group of protozoa, most of which possess a unique organelle called apicoplast and an apical complex structure involved in penetrating a host's cell. They are unicellular, spore-forming, and exclusively (Jadwiga, 1991) parasites of animals. Motile structures such as flagella or pseudopods are absent except in certain gamete stages. This is a diverse group including organisms such as coccidia, gregarines, piroplasms, haemogregarines, and plasmodia. 3 1.4.1. Haemogregarina spp Four genera of intracellular parasites are included among the Haemogregarines; Haemogregarina, Hepatozoon, Karyolysus, and Hemolivie. These genera cannot be accurately classified based on their appearance in blood cells alone (Al-Farraj, 2008). On blood films, the gamonts of Haemogregarines appear as sasusage-shaped inclusion with pale to purple cytoplasm and one centrally to slightly eccentrically placed, darker purple staining nucleus except in Haemogregarines infection where erythrocytic meronts may be present. These are unpigmented and are typically found of the cytoplasm of red cells and sometimes in white blood cells (Jacobson, 1983). The gamonts may push the nucleus of the host cells to inside or surround it. The hosts cells may appear irregular in shaped and size (Shazly, 2000). Rarely, two or more organisms may be found in one erythrocyte, or the gamonts may be found extracellularly. Because gamonts of different Haemogregarines are morphologically indistinguishable in the peripheral blood, the general terms Haemogregarines is used to report their presence. Haemogregarines belonging to the genus Hepatozoon are commonly found in terrestrial and aquatic snakes. Haemogregarines sporozoites are often transmitted by infected arthropods and leeches. Haemogregarines protozoa are well adapted to their natural host, but can cause significant clinical inflammatory disease in unnatural host species (Keymer, 1981). 1.4.2. Plasmodium spp Five genera of the family plasmodiidae are reported: plasmodium, Fallisia, Saurocytozoon, Haemocytstidium, and Haemoproteus. Snakes and turtles may become infected with Hemoprotus. Whereas plasmodium, Fallisa, 4 Saurocytozoon, and Haemocytstidium infections have been reported in lizards (Lane and Mader, 1996). 1.5. Microfilaria spp Various genera of filaridae worms can be found in reptiles. Some of which are specific, such as Macdoualdius spp which have been detected in some species of snake and lizards, Saurositus spp which have been detected in lacertid lizards, Foleyella spp which have been detected in some species of chameleons lizards, and cardinema spp which have been detected in chelonians lizards (Lane & Mader, 1996). Microfilariae are transmitted by blood sucking mosquitoes or ticks or some sandfly species such as Phlebotomus duboscqi. Filariasis in reptiles is mostly subclinical, but with heavy infestation, thrombosis and blockage of blood vessels may occur resulting in edema, fibrosis and/or necrosis in the affective area (Lane & Mader, 1996). 1.6. Zoonotic diseases of reptiles and amphibian There are several diseases which can be transmitted from reptiles and amphibain to humans such as Salmonellosis, Tuberculosis, also many reptiles can harbour some protozoan organisms capable of causing diseases such as Cryptosporidiosis which is caused by a coccidian protozoan (Kolle &Hoffmann, 1998). Rationale: Due to the lack of knowledge regarding the zoonoses diseases of reptiles and amphibian in the Sudan, there is a great need to identify those parasites firstly in order to know their zoonoses importance. 5 Objectives: Thus the present study was conducted to carry out the following: Survey of different blood parasites in reptiles and amphibians Identify types of blood parasites in two lizards’ species (Mabuya striata, Mobuya quinquetaeniatus) and toad (Bufo regularis). Comparison of blood parasites in reptiles and amphibians according to microscopic diagnosis. 6 Chapter two Material and methods 2.1. Study area The samples were collected from two locations in Khartoum state; Tuti Island which lies in the union of the Blue and the White Nile and Jebel Awlyia area near the White Nile in Khartoum south (Fig.1). 2.2. Sample collection and preparation Thirteen lizards’ specimens from species Mabuya striata and two specimens from species Mobuya quinquetaeniatus from Tuti Island (Blue Nile), and 15 toads’ specimens from species Bufo regularis from Jebel Awlyia area (White Nile) were captured during this study. The animals were dissected and tissue samples were obtained from spleen and liver and preserved in eppendorf tube in Isopropanol alcohol 95%. Blood samples were taken, blotted on filter paper (Whattman No.3) and air dried, then preserved in separated plastic bags. 2.3. Microscopic examination for tissue samples Tissue from spleen and liver were spotted on slides, fixed with absolute methanol, air dried, stained with Giemsa, and examined under light microscope with oil emersion lens to search for parasites. Then the positive slides were photographed using a microscope digital camera (DCE-2). 7 Fig.1: Satellite image showing the location of the two study areas in Khartoum. 8 2.4. Identification of the parasites The identification for microfilaria nematodes were done according to the characteristics described by Bain, et al., (1992), identification for Sauro-
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