Authors

A NIKAM¹; S Balogun¹; AL Webb¹; O Polak¹; Z Krupa¹; B Lesetedi¹; M Buerdsell¹; B Davies¹; C Evans²; J King²; J Alexander¹; JA Pachebat¹;  ¹ Aberystwyth University, UK;  ² Wales Veterinary Science Centre, UK

Discussion

Neonatal diarrhoea is caused by the protozoan parasite Cryptosporidium species. Young animals are particularly vulnerable to the parasite, and they might experience severe diarrhoea and a high mortality rate, which results in large economic losses from mortality, anorexia, and stunted growth. Several Cryptosporidium species are known to infect neonates, with different pathogenicity. The diagnostic identification of oocysts under a microscope is not very reliable because oocysts from different Cryptosporidium species share similar morphology. Molecular detection involves PCR followed by restriction fragment length polymorphism (RFLP) or gene sequencing and is expensive and time-consuming. Most investigations have only been done on a small quantity of positive clinical samples and are either genus wide or target a specific species making it challenging to accurately determine the incidence of Cryptosporidium species present in animals and to specifically identify the species causing disease. To address this issue, a multiplex PCR assay is being developed and optimised to identify several Cryptosporidium species in sheep and cattle faecal samples, with the aim of characterising PCR amplicons by nanopore sequencing. With regard to current sequencing techniques, this improvised method for mixed infection diagnosis saves time and money, as the only reliable way of identification of mixed infection samples with high specificity and sensitivity.