Inverstigation And Species Identification On Parasites From Three Species Of Snakes

Snake in the process of long-term animal evolution, formed as a reptile, is on^-ofthe important part of biodiversity. Snakes in maintaining natural ecological balance,have the important and irreplaceable role, and is closely related to human health. Theexistence of snake parasites, seriously affected the snake’s growth, to reduce the snakebody，lead to snakes are more likely to infect other disease, serious when still canmake the snake died. So investigation and identification of the type of snake parasite,have a vital role to the protection of snakes.1. Some snakes parasite infection situation preliminary investigation of ShanghaiTo investigate the infective status of parasites in snakes from Shanghai,29snakes from3species (9Dinodon rufozonatum,10Elaphe carinata and10Elaphetaeniura) confiscated from the market were dissected. The viscera (body,subcutaneous，muscles, heart, lungs, liver, stomach, intestinal organs) and bloodsmears of the snakes were examined. The parasites from these viscera were collectedand observed by microscope. The results showed nematodes, tapeworms andHepatozoon were found, but no ectoparasites, trematodes and acanthocephalan. Theparasitic infection rate of snakes checked was96.55%. The infection rate ofnematodes, tapeworms and Hepatozoon were77.88%,100%,0in Dinodonrufozonatum,100%,100%,100%in Elaphe carinata and60%，40%，0in Elaphetaeniura respectively. A total of241nematodes and1260tapeworms were collected in29snakes, and69.79%of nematodes and86.55%of tapeworms were from Elaphecarinata. According to the viscera,93.20%of tapeworms were found in subcutaneousand65.63%of nematodes in stomach. The results indicated the parasitic infection rateand intensity of snakes from Shanghai market were very high. Sparganum mansonifound in this investigation was zoonotic parasite, and it is easy to infect humansthrough eating snake skin, meat and gall. Therefore, protecting wild animals likesnakes is also to protect human themselves. 2. Morphologic observation and species identification of HepatozoonHepatozoon species are the most common hemoparasites of snakes. The resultsshowed that10Elaphe carinata are infected Hepatozoon. The snakes were naturallyinfected. Blood smears were prepared immediately upon collection and the remainingblood was rfozen at-20°C for molecular characterization studies. All ten Elaphecarinata were found to include intraerythrocytic parasites and each of them infectedwith one gamont type. Mature gamonts were slender with a round posterior end andanterior extremity that tapered slightly to a blunt point and folded back in a hook-wisefashion for about3fxm. They measured15.25土0.76x2.45土0.59|xm (13.24-17.25x1.12-4.1, n=188) in size with an area of43.92±9.252\im (20.48-68.77, n=188). Thenucleus was elongated, compact and always present in second quarter of gamont,usually extending into first and third quarter. They measured6.92士1.26x2.02±1.18jim (4.13-10.06x1.08-3.22) with an area of12.99土4.082\im (5.49-22.85).Double erythrocyte infections were also detected. For molecular characterization,HepF300/Hep900and HEMOl/HEM02were used to amplify and distinguishdifferent isolates of Hepatozoon spp. from snakes. The topology of the tree indicatesthat sister taxa to this lineage appears as Hepatozoon sp. from Bandicota indica and H.ayorgbor from Python regius. But they have obvious differences on the form. In thisstudy，the gamont folded back hookwise for about3p,m, this phenomenon has notbeen reported. Based on the molecular and morphologic, a new species was proposed:Hepatozoon chinensis n. sp. This is the first description of Hepatozoon species fromsnakes, based on molecular characterization and morphological data in China.3. Morphologic observation and species identification of Plerocercoid fromsnakesPlerocercoid is Diphyllobothriidea tapeworm the floorboard of the larva. Seizedby figuring out the the kinds of Plerocercoid, this study using the methods ofmorphology and molecular biology, to identification of the Plerocercoid. The resultshows: the Plerocercoid was milk-white, about0.68?5.18mm long, for long stripentity structure. Plerocercoid begin to end is wider, with head quarter of the adult Plerocercoid, and thinner. Species development tree analysis showed that within thegroup can be divided into3big teams, the ifrst team is composed of overlappingpalace is a tapeworm, the survey of the Plerocercoid in the group, and with previouslyreported Spirometra erinaceieuropaei in the same branch. Such a grouping, conformto the natural species isolation conditions, and the in the evolutionary tree，thebootstrap value is very high, shows the result is reliable. So Plerocercoid found in thissurvey can be identified as Spirometra erinaceieuropaei, consistent withmorphological analysis.4. Morphologic observation and species identification of Nematodes from snakesThis study deals with the method of morphology and molecular biology of seizedsnakes nematodes, verification results show that:(1) Ascarid, parasites in the stomachor intestines. In the gut of specimen body tend to form a smooth dough, often inintestinal parasitic in the lining of the stomach of the insect body arranged in line,then the worm’s head and tail again out of inner cavity of the stomach, while theworm in the stomach, specimen body are tan or iron gray, head end there are three lips,lips have a row of serrated teeth, no intermediate lip, no neck wing membranes. Malemeasured length54.03?83.56mm, wide0.80?1.37mm. The esophagus long3.40-4.68mm, The tail end with a small thorn, two root, such as long slenderspiculum, length of about1.27mm. The tail distribution measurement has two beforea row of anal papilla. Female is relatively larger than males, length70.36?100.08mm,and wide1.02?1.49mm. Esophagus was also longer than male,3.85-9.55mm.. Vulvain the back-end, specimen head to the distance of the vulva accounted for54.60%?61.10%of specimen body length. And consistent to literature of Ophidascaris iflariadescribe.(2) Kalicephalus: specimen have slender body, small mouth capsule, straightforward, cutin ridge is wide, before atfer abdominal Angle square, dorsal horn atfer anarc, back channel base wide, back groove ridge consists of moniliform cutin piece.The esophagus blunt, the back of the significant enlargement, nerve ring located in thefront of the esophagus, drain openings in the base of the esophagus, neck mastoid islocated in the rear of the drain hole. The specimen was consistent the literature description of Kalicephalus indicus. According to the morphological identification,the survey can be seized by the Ascarid identified as Ophidascaris iflaria, andKalicephalus identified as Kalicephalus indicus. Lack of snake nematodes molecularappraisal reports, so want to pass on the phylogenetic tree analysis and identificationof snake nematode species，it seem Impossible. So the molecular identification ofsnake’s nematode needs further research.