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The present study was aimed to evaluate the diagnostic value of enzyme linked immunosorbent assay (ELISA) using excretory-secretory antigen (ESA) and Zinc sulphate sedimentation technique for coprodiagnosis. A comparison was made between both techniques for rapid and accurate diagnosis based on 200 serum and faecal samples from cattle (n=117) and buffaloes (n=83). In the first step, ELISA was standardized by using sera from Fasciola infected and non-infected animals. The level of sensitivity observed was 100%, while 95% specificity against Gigantocotyle explanatum and Paramphistomum spp. or a mixed infection with both parasites was noted. Fasciolosis rate was significantly (P<0.05) higher in buffaloes in which 60% prevalence was found for indirect ELISA and 53% in case of coprological analysis, while on other hand in cattle prevalence rate was 50 and 37%, respectively. The results showed a significant (P<0.05) breed difference in cattle. Furthermore, the significant difference was noted in buffaloes with reference to age and sex groups. After contrasting data from ELISA and faecal analysis, 11% of the sera analyzed had positive values of indirect-ELISA and negative by faecal analysis. The findings suggest that indirect ELISA is an efficient diagnostic technique and combination of both techniques could be useful for accurate diagnosis of fascioliasis in endemic areas. It is recommended that both cattle and buffaloes should be screened by applying both serological and coprological techniques in other agro-ecological zones of Pakistan. [ABSTRACT FROM AUTHOR]

In present study, 50 samples representing Fasciola (Platyhelminthes: Trematoda: Digenea) from cattle and buffaloes of Pothwar region were characterized genetically by sequences of the first (ITS-1) and second (ITS-2) internal transcribed spacers (ITS) of nuclear ribosomal DNA (rDNA). The ITS rDNA was amplified from individual liver flukes by polymerase chain reaction (PCR), and the amplicons were sequenced directly. The lengths of the FG ITS-1 and ITS-2 sequences were 391 and 329 bp, respectively, for all Fasciola gigantica samples sequenced. For Fasciola hepatica samples sequenced, FH ITS-1 was not amplified and length for ITS-2 sequences was 330 bp. Molecular identification using rDNA internal transcribed spacer 1 and 2 characterizes Fasciola spp. in Pothwar to be an intermediate/ hybrid resembling more with F. gigantica. The results also indicated that Fasciola spp in cattle and buffalo are genetically similar, as both hosts' possess Fasciola with the same number of base pairs in their internal transcribed spacers as proved by gel electrophoresis and sequencing. To determine the phylogenic location of Fasciola spp. of Pakistan based on rDNA molecular data, ribosomal ITS regions were sequenced and compared with other fasciolid species from NCBI databases for sequence homology analysis using BLAST and ClustalW programs. The phylogenetic trees revealed a close relationship with isolates of F. gigantica from China, India, Iran and Vietnam. This is the first demonstration of the existence of hybrid or an intermediate Fasciola species in Pakistan by a genetic approach, which provides foundation for further studies in different agro ecological zones of Pakistan. [ABSTRACT FROM AUTHOR]

The article discusses a research study on the taxonomic position of adult Fasciola spp. that infect cattle and buffalo in the Potohar Region of Punjab in Pakistan. Study subjects include 54 adult gravid flukes gathered from 7 cattle and 153 bufffalo and processed for staining with Gower's carmine then morphological measurements were taken and the biometric parameters for phenotypic analysis were considered including the length cone (CL), maximum width of body (BW) and maximum diameter of oral sucker (OS max). Morphometric analysis showed that measurements of nine paremeters were resembling that of the F. gigantica while 6 parameters resemble Fasciola spp and 5 parameters resembling F. hepatica. Also noted is the use of cluster analysis in the hierarchical approach.