458 Asin P J Trop Dis 2017; 7(8): 458-462 Asin Pifi Journl of Tropil Disese journl homepge: http://www.pjtm.om Originl rtile https://doi.org/10.12980/pjtd.7.2017d7-48 2017 y the Asin Pifi Journl of Tropil Disese. All rights reserved. A study on endoprsiti nd etoprsiti fun of snkes in Mizorm, Indi Gutm Ptr 1*, Sonjoy Kumr Borthkur 1, Seikh Shnwz Alm 2, Suhmoy Ghosh 1, Hmr Lllinkimi 3, Hning Llrinkim 1 1 Deprtment of Veterinry Prsitology, College of Veterinry Sienes nd Animl Husndry, Selesih, Aizwl, Indi 2 Deprtment of Veterinry Miroiology, West Bengl University of Animl & Fishery Sienes, K.B. Srni, Kolkt, W.B., Indi 3 Deprtment of Livestok Prodution & Mngement, College of Veterinry Sienes nd Animl Husndry, Selesih, Aizwl, Indi ARTICLE INFO Artile history: Reeived 7 Mr 2017 Reeived in revised form 9 My 2017 Aepted 18 Jun 2017 Aville online 4 Aug 2017 Keywords: Endoprsites Etoprsites Snkes SEM Mizorm Indi ABSTRACT Ojetive: To reord the prevlene of prsiti fun of snkes from different prts of Mizorm, Indi. Methods: Colleted fel smples of different snkes were exmined y sedimenttion nd flottion tehniques. Similrly, lood smples were exmined for presene of ny hemoprotozo following Giems stining tehnique. Etoprsites were identified on the sis of morphologil keys. Snning eletron mirosopy (SEM) ws performed for detiled surfe struture studies of few prsites. Results: Sixty one perent (40/65) snkes exmined were found positive for prsiti infetion. The predominnt endoprsites inluded five speies of nemtodes, one speies of estodes, one speies of tissue protozo, nd two speies of hemoprotozo. Etoprsites reorded were tiks of the gener Aponomm nd Amlyomm. The most undnt nemtode reorded ws Kliephlus speies. Conlusions: The study indites tht prsiti infetion of snkes is quite ommon in this prt of Indi nd deserves ttention for zoologil studies. 1. Introdution Reptile plys signifint role in the eosystem. They eome one of the ommon pet nimls throughout the world. Like other speies, they n lso hrour vrious prsites[1-7]. In zoos, reptilin speies inluding snkes re often losely onfined together nd this uses stress resulting in ltertion of host-prsite reltionship. Sometimes reptile n lso e responsile for trnsmitting disese to nimls nd humns. The reptiles hror prsites for onsiderle period of time without showing ny oservle symptoms unless other ftors undermine the host immunity nd llow the disese to develop[8]. For veterinrins or zookeepers, urte identifition of prsites nd their stges is ruil for helth of reptiles. Aprt from spordi reports on prsiti infetions of reptiles from different prts of Indi[5-7], there is limited informtion on prevlene of prsites in snkes. In view of the ove mentioned, the present study ws onduted *Corresponding uthor: Gutm Ptr, Deprtment of Veterinry Prsitology, College of Veterinry Sienes nd Animl Husndry, Selesih, Aizwl, Indi. Tel: +91 8582859415 E-mil: dr.gutmptr@yhoo.o.in All experimentl proedures were onduted ording to Institutionl Animl Ethi Committee with referene No. CVS/CAU/IAEC/no. 6641, dtd, Selesih, the 25th, April, 2016. Foundtion Projet: Supported y Centrl Agriulturl University, Imphl, Mnipur, Indi y grnting ollege reserh funding support (sntion order No. IRP/1921/NE/2016, dtd, the My, 31, 2016). The journl implements doule-lind peer review prtied y speilly invited interntionl editoril ord memers. to reord endoprsites nd etoprsites of snkes y vrious methods. Snning eletron mirosopy (SEM) ws lso pplied for detiled surfe strutures of few prsites. 2. Mterils nd methods 2.1. Ple of study Study re ws onfined to different prts of Aizwl Distrit of Mizorm nd Aizwl Zoo, Mizorm, where snkes re kept for puli view. The survey ws rried out from Otoer, 2016 to Jnury 2017. Further, few snkes run over y vehiles or killed y people were lso inluded in this survey. A totl of 65 snkes elonging to the fmilies Pythonide, Viperide, Elpide, nd Coluride were exmined. 2.2. Colletion nd exmintion of fel smples Fresh fees (n = 235) were olleted from ptive snkes s prtile s possile. Diret smer, flottion nd sedimenttion tehniques were used for fel exmintion s per stndrd proedure[9-12]. Fel smples of ded snkes were exmined during post mortem exmintion. 2.3. Exmintion of lood smples Blood smples were olleted from few live snkes t ptivity s well s from the reently killed snkes nd stined with Giems stin
Gutm Ptr et l./asin P J Trop Dis 2017; 7(8): 458-462 459 s per proedure nd exmined under oil immersion mirosope. 2.4. Postmortem exmintion At lest 30 ded snkes were rought to the Deprtment of Prsitology during the whole study period nd postmortem exmintions were onduted. Snkes were ut open in the lortory, nd viser were exmined nd ny prsites reovered were kept in 70% lohol for further study. 2.5. Etoprsite survey Skin of live or ded snkes ws refully exmined for presene of ny etoprsites. Etoprsites found were lso stored in 70% lohol for further study. Bodies of the tiks were perforted with entomologil pin efore oiling with 10% potssium hydroxide for 5 10 min. The treted smples were then pled in 70% lohol for 5 min nd 90% lohol for nother 5 min for dehydrtion. Finlly, the speimens were kept in roli id for 10 min efore mounting in DPX. 2.6. Preprtion of smple for SEM Nemtodes were fixed in 3% glutrldehyde. Before dehydrtion, ny unwnted mterils like fel deris, muus, lood or other ody fluids were refully removed y wshing severl times with nulese free wter (NFW) with the help of mel rush. The smples were then fixed t 4 C for 24 h. The fixed nemtodes were wshed in phosphte uffered sline (ph 7.2) for three times nd then in doule distilled wter followed y etone dehydrtion. After etone dehydrtion, the speimens were dried with liquid CO 2 t its ruil point i.e. 31.5 C t 1 100 psi. The speimen ws then dipped in tetr methyl sline (TMS) for 5 10 min with two hnges t 4 C. They were then dried t room temperture (25 26 C ). The smples were then mounted on luminium stus. Finlly the smples were oted with gold sputter nd oserved under SEM (JSL 6300 JEOL) t sophistited nlytil instrument filities lortory t North Estern Hill Univeristy (NEHU), Shillong, Meghly, Indi. 2.7. Ethil sttement All experimentl proedures were onduted ording to Institutionl Animl Ethi Committee with referene No. CVS/ CAU/IAEC/no. 6641, dtd, Selesih, the 25th, April, 2016. 3. Results A totl of 65 snkes elonging to 5 fmilies (Pythonide, Coluride, Elpide nd Viperide) were exmined nd the results of the fel smples nd lood smples re shown in Tle 1. Thirty snkes (46.15%) were found to e infeted with six different speies of nemtodes nd one speies of estode. Two snke fel smples were positive for Isospor sp. nd seven snkes were found infeted with two speies of hemoprotozons, nmely, Heptozoon sp. nd Trypnosom sp. Tle 1 Endoprsites of snkes. Endoprsites Venomous Non-venomous (n = 35) (n = 30) Nemtodes Kliephlus sp. 10 3 Ophidsris sp. 5 2 Polydelphis sp. 0 2 Gnthostom sp. 1 0 Tng tir 1 0 Strongylus sp. 2 2 Cestodes Ophioteni sp. 1 1 Tissue protozo Isospor sp. 1 1 Hemoprotozo Heptozoon sp. 2 3 Trypnosom sp. 2 0 Results of the etoprsiti fun of different snkes re shown in Tle 2. Nine snkes (13.84%) were positive for tiks Aponomm vrnense nd 5 (7.69%) positive for Amlyomm gervisi. Tle 2 Etoprsites of snkes. Etoprsites Speies of snkes P. molurus King or Green viper Ptys muos (n = 20) (n = 20) (n = 15) (n = 10) Aponomm vrnense 4 3 1 1 Amlyomm gervisi 1 2 1 1 Bsed on egg nd morphology under light nd eletron mirosopy, five speies of nemtodes were identified s Kliephlus sp. (Figure 1), Ophidsris sp. (Figure 2), Polydelphis sp. (Figure 3), Strongylus sp. (Figure 4) nd Gnthostom sp. (Figures 5 nd 6). Further, tpeworm ov (Figure 7) were deteted only in rt snkes (Ptyts muos) nd isosporn ooysts deteted in two fel smples (Figure 8). Five snkes (one python, two green vipers nd two king ors) were found positive for Heptozoon sp.; on the other hnd, two venomous snkes were found positive for Trypnosom sp. (Figure 9). Light mirosopil (LM) nd SEM pitures of Aponomm vrnense nd Amlyomm gervisi re shown in Figure 10. Figure 1. Egg of Kliephlus sp. (40 ) (), nterior prt of Kliephlus sp. () nd posterior prt of Kliephlus sp. ().
460 Gutm Ptr et l./asin P J Trop Dis 2017; 7(8): 458-462 200 100 μm Figure 2. Adult Ophidsris sp. from ded python (), egg of Ophidsris sp. () nd nterior prt of Ophidsris sp. () under SEM. 150 100 μm 95 200 μm Figure 3. Egg of Polydelphis sp. (), nterior end of Polydelphis sp. () nd posterior end of mle Polydelphis sp. (). Figure 6. Posterior end of femle Gnthostom sp. Figure 4. Egg of Strongylus. 95 200 μm 150 100 μm Figure 5. Anterior end of Gnthostom sp. Figure 7. Egg of tpeworm (10 ).
Gutm Ptr et l./asin P J Trop Dis 2017; 7(8): 458-462 461 4. Disussion Figure 8. Sporulted ooyst of Isospor sp. Figure 9. Heptozoon sp. () nd Trypnosom sp. () in the lood of snke. The present study ws undertken to reord the seline dt of etoprsiti nd endoprsiti fun of snkes of different prts of Mizorm, Indi. Prsites my hve detrimentl effets on the well-eing of reptiles kept in zoos, frms or s domesti pets[4,5]. The different tehniques were used for detetion of prsiti ov in fees of snkes s per previous reserhers[9-12]. For the identifition of nemtode eggs nd protozon ooysts, fel flottion ws found superior to other methods. Eggs nd ooysts were identified sed on morphometry. The result indited high prevlene of Kliephlus sp. (20%) whih greed with the finding of previous reports[13,14]. The speies of Kliephlus is hrterized y strongly developed ul psule with three teeth like ppille nd well developed oesophgus with ulous posterior end. The high prevlene of Kliephlus sp. my e due to su-optiml onditions of the ptivity, lk of deworming prties in zoo s well s different feeding hits of snkes. At postmortem exmintions, dult worms were identified on the sis of morphology nd ov were deteted from the gut ontents. Asrid eggs nd oidin ooysts s well s tpeworm were lso deteted. Asrid eggs were spheril to su-spheril in shpe with rownish yellow shell, strited nd were out 5 6 µm in thikness. The prevlene rtes of srid in poisonous nd non-poisonous snkes were found similr. Ophidsris ws more ommon in venomous snkes, while Polydelphis ws more undnt in non-poisonous snkes. Severl speies of the fmily Asridide inluding Ophidsris nd Polydelphis hve een reported[15-17]. De Souz et l.[18] nd Bino Sundr et l.[19] lso reported Ophidsris in snkes. Eggs of Ophidsris nd Polydelphis were found in the fel smples from pythons, ors, green vipers nd rt snkes. Out of 26 venomous snkes, 10 were found to hve mixed infetions. On the other hnd, out of 14 nonvenomous snkes, 4 hd mixed infetions. Only one rt snke ws found positive for estode infetion. This is in ontrst to the previous report y Rjesh et l.[11] who filed to reord ny tpeworm infetion. In the present investigtion, Gnthostom infetion in one Indin or ws reorded. The prsite ws found morphologilly similr to the speies Tngr tir oserved y Chnhome nd Chiyutr[13]. Two hemoprotozon prsites nmely Heptozoon sp. nd Trypnosom sp. were lso oserved in this present study. Smith[20] reported tht Heptozoon speies re the most frequently 100 100 μm 170 100 μm Figure 10. Amlyomm gervisi (), nterior prt of Amlyomm gervisi under SEM () nd mouth prts of Aponomm vrnense () under SEM.
462 Gutm Ptr et l./asin P J Trop Dis 2017; 7(8): 458-462 ourred hemogregrins in different speies of snkes. Two venomous snkes nd three non-venomous snkes were found positive for Heptozoon sp. Two venomous snkes were lso found positive for Trypnosom sp. Lower level of oidin infetions ws found in the present study. Aponomm vrnense nd Amlyomm gervisi were the only two gener of tiks reovered from 14 snkes in the present study. The Aponomm vrnense ws identified on the sis of keys provided y Hoogstrl et l.[21]. Burridge[22] reported tht Aponomm vrnense is one of the most widespred speies of the genus Aponomm, ourring in snkes of mny ountries inluding Indi. Hork et l.[23] synonymized Aponomm nd Amlyomm under the sme genus Amlyomm; however, mny sientists hve not epted this modifition. Amlyomm gervisi ws found more undnt in poisonous snkes thn in non-poisonous one. Etoprsites, espeilly tiks, re frequently seen in wild reptiles ut the level of infetion nd the effets of tiks on the snkes re poorly studied[24]. Although tik rrely ours in lrge numers in snkes to use diret injuries, they n t s rriers of vetor orne diseses to other nimls nd even to humns. The findings indited tht mny snkes hroured etoprsites or endoprsites whih my e due to severl ftors tht re omplementry to eh other suh s feeding hits, environmentl onditions, mngement prties, immune defiieny, et. Hene, it is worthy to exmine linil smples for helth sttus of snkes under ptivity. Furthermore, ntiprsiti tretment nd etter mngement ensure the helth nd well eing of snkes. Conflit of interest sttement We delre tht we hve no onflit of interest. Aknowledgments The uthors re thnkful to the Den, College of Veterinry Sienes nd Animl Husndry, Centrl Agriulturl University, Selesih, Aizwl, Indi for providing neessry filities for onduting the experiment. Referenes [1] Al-Mousswi AA. 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