TDK conference 2019

Haemonchus contortus, the parasite species which lives in the abomasum, causes important animal health and economical damage in in sheep and goat flocks worldwide. In the last decade the parasitosis got to the centre of attention in Europe related to climate change and the fact that resistance to anthelmintics is becoming more common. The presence of H. contortus in Hungary has long been known, but there is still no data about its prevalence, animal health and economical importance in the local sheep flocks. The infection of this nematode species cannot be diagnosed by eggs, the culture and morphological examination of 3rd stage larvae are time-consuming and they require considerable experience. Besides the nationwide questionnaire survey, the occurrence of the parasite was examined in sheep flocks with two types of molecular biological methods first time in Hungary.

The presence of haemonchosis was mentioned in six out of 911 complete questionnaires, and more owners observed the typical clinical signs of parasitosis such as anaemia and submandibular oedema. Pooled faecal samples were collected once in 37 flocks of 14 counties. Strongylid type eggs of gastrointestinal parasites were examined with mini-FLOTAC method. This kind of egg was found in all samples, except one. Eggs were washed out from the samples of 10 flocks, and DNA was extracted with NucleoSpin Tissue© (Macherey-Nagel) kit. H. contortus was detected with conventional PCR and another amplification method called Loop-mediated isothermal amplification (LAMP). In PCR reaction the extracted DNA of adult worms were used as positive control. In LAMP the positive control DNA and PCR water as negative control were used. The products obtained with PCR were sequenced. Altogether this nematode species was detected in six flocks in six counties, where the presence of H. contortus had not been known earlier.

The results of the questionnaire survey and the molecular biological methods contribute to our knowledge of this important parasitosis. The presence of H. contortus in a flock can be detected with LAMP quickly. This method is 10 times more sensitive than PCR, the result is reliable even if faecal egg count is only 2 eggs in one gram faeces.