000			04241nam a2200181Ia 4500
005			20151006142314.0
008			150525s2013 xx 000 0 und d
041			_aeng
082			_a1681,T
100			_aAli Abbas _98427
110			_cDr. Muhammad Avais _95199
245			_aPrevalence And Molecular Genetic Characterization Of Different Isolates Of Hydatid Cyst In Ruminants In Okara
260			_c2013
502			_aCystic hydatid disease has a worldwide distribution. Echinococcusis cause great economic loses in Pakistan and lead to the loss of worth 276.20$ per 100 sheep and goats, as well as 165.72$ per 100 infected buffaloes, cattle and camels. It has zoonotic importance and also have well-recognized zoonosis in Pakistan and numerous cases have been reported in the medical literature. Cystic hydatid disease is caused by Echinococcus granulosus, (E. granulosus), tapeworms. Adult tapeworms of this specie are small in size. Their measurement revealed that they reach upto 2-11 mm in length and contain protoscolex, the cephalic end for attchment and 2-5 proglottids segments. Two rows of keratinized hooks and four number of suckers are present on scolex. The last gravid proglottids segments bear the large number of fertilized eggs, which are shed after every 7-14 days. Eggs are oval in shape, having clearly distinct oncophore and 30-36 micrometer in diameter. There are several different strains of E. granulosus, which are genetically distinct. These strains vary phenotypically and therefore, this feature can be used for the control of this parasite. 10 genotypes of E. granulosus are identified up till now. Therefore, in this study we used PCR technique for prevalence determination and molecular characterization of specific strain of E. granulosus. The primers specific for E. granulosus were used in this study. The E.g.ss1for (5¡Ç-GTA TTT TGT AAA GTT GTT CTA-3¡Ç) worked as forward primer, while E.g.ss1rev (5¡Ç-CTA AAT CAC ATC ATC TTA CAA T-3¡Ç) worked as reverse primer. For this purpose, total 200 numbers of cyst samples (100 from each district) were collected from liver and lungs of cattle, buffaloes, goats and sheep slaughtered at different private and public abattoirs in Okara and Jhang. An antimortem examination was performed on each animal LIV and data regarding each animal entered in data capturing form before slaughter. Whole cyst sample was collected without rupturing and preserved in ice packs. Then these isolates were transferred to laboratory and stored at -20 ¢ªC for further processing. For the genetic analysis of E. granuous (Hydatid cyst) DNA was extracted from germinal layer and cystic fluid by using DNA extraction reagent (TRIREAGENT¢ç, Molecular Research Center, Ohio, USA) according to the manufacturer.s instructions. PCR was carried out by using primers specific for G1 strain of E. granulosus. The whole three steps (Denaturation, Anealing, Amplification) of PCR was carried out in PCR thermo cycler under conditions specific for E. granulosus. The final PCR product was electrophoresed in a 1% agarose gel containing 0.5 ug/ml of ethedium bromide. Electrophoresis was completed by applying 90 volts for 40 min. After required time, gel was placed in UV trans-illuminator to visualize the band shown by specific genotype of E. granulosus. Specific bands at 254 bp confirmed the G1 strain of E. granulosus. Results indicated the prevalence of 37% in Okara and 65% in Jhang district of Punjab. While in cattle (58.00%), buffalo (76.00%), sheep (36.00%) and goats (34.00%) prevalence was observed. As well as, (44.7%) prevalence of E. granulosus was calculated in liver and (54.8%) was observed in lungs in both districts. So, this study predicted the prevalence of hydatid cystic disease in ruminants in Okara and Jhang. PCR technique was used for the diagnosis of hydatid cyst in animals. This molecular characterization technique enables us to know the specific strain of E. granulous existing in these two districts. Finally the data gathered from this study help to understand the disease structure and to develop future plan.
650			_aDepartment of Clinical Medicine & Surgery _94563
700			_aProf. Dr. Muhammad Sarwar Khan _95200
942			_cTH
999			_c3389 _d3389