Abstract
Haemonchus contortus commonly called the stomach worm or wire worm of ruminants inhabits the abomasum and is considered to be one of the economically important gastrointestinal strongyles in goats. In the present study, H. contortus was identified by PCR using the primers targeting partial 5.8S and partial internal transcribed spacer region 2 (ITS-2). Adult worms were identified morphologically and genomic DNA was extracted using DNeasy Blood and Tissue kit (QIAGEN, Germany). Gradient PCR protocol was standardised using the extracted genomic DNA. Ten-fold serial dilution of adult DNA was used to analyse the minimum detection limit and the products were amplified upto the tenth dilution. Cross reaction of primer sets was checked using the DNA extracted from predominant adult srongyles like Oesophagostomum columbianum and Trichostrongylus colubriformis and no cross reaction was seen at the optimum annealing temperature (60.7°C).
Highlights
Haemonchus contortus commonly called the stomach worm or wire worm of ruminants inhabits the abomasum and is considered to be one of the economically important gastrointestinal strongyles in goats
Gradient polymerase chain reaction was standardised with initial denaturation at 94 oC for 5min followed by 39 cycles of denaturation (94 oC for 30s), annealing (57 to 63°C for 30s), extension ( 72 oC for 30s) and final extension at 72oC for 10min
After performing gradient PCR, the amplicons were subjected to agarose gel electrophoresis in 1.5 per cent agarose gel at 80V, 400mA for 35 min and the gel was visualised in Gel Doc TM EZ imager and documented using Image lab software
Summary
Haemonchus contortus commonly called the stomach worm or wire worm of ruminants inhabits the abomasum and is considered to be one of the economically important gastrointestinal strongyles in goats. Adult worms are identified based on morphological features. Identification of nematode species based on features of the strongyle egg is difficult during coprological examination. Coproculture aids in species identification but it takes seven to ten days to identify the infective larvae.
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