First nationwide investigation of Cryptosporidium species and gp60 subtypes in dairy cattle in Cyprus

Cryptosporidium spp. are globally important enteric pathogens in livestock and a leading cause of neonatal calf diarrhoea, with zoonotic potential. This study presents the first nationwide molecular survey of Cryptosporidium spp. and C. parvum gp60 subtypes in dairy cattle in Cyprus. A total of 517 faecal samples were collected from dams (n = 256) and their 3–7-day-old calves (n = 261) across 18 farms in five districts. Samples were screened by nested PCR and sequencing of the SSU rRNA and gp60 genes. Overall, Cryptosporidium spp. prevalence was 28.2%, with higher rates in calves (39.5%) than in dams (16.8%). Cryptosporidium parvum was the dominant species (85.6%), followed by C. bovis (8.2%), C. andersoni (4.8%), and C. ryanae (0.7%). Eight C. parvum gp60 subtypes were identified, six from family IIa and two from IId, the latter reported for the first time in Cypriot cattle. Subtypes IIaA14G1R1 and IIdA16G1 were strongly associated with severe diarrhoea, while IIaA17G2R1 predominated in asymptomatic calves. Several identified subtypes have been implicated in human outbreaks globally. Cyprus’s confined livestock population and strategic location at the crossroads of Europe, Asia, and Africa provide a unique context for understanding Cryptosporidium transmission. This study establishes essential baseline data on the prevalence, molecular diversity, and zoonotic potential of Cryptosporidium in an island setting. The findings underscore the need for integrated surveillance and One Health strategies to control transmission, protect public health, and monitor the emergence of high-risk subtypes locally and beyond.