Cross-Border Investigations on the Prevalence and Transmission Dynamics of Cryptosporidium Species in Dairy Cattle Farms in Western Mainland Europe

Abstract

Cryptosporidium is an apicomplexan parasitic protist, which infects a wide range of hosts, causing cryptosporidiosis disease. In farms, the incidence of this disease is high in animals such as cows, leading to extensive economic loss in the livestock industry. Infected cows may also act as a major reservoir of Cryptosporidium spp., in particular C. parvum, the most common cause of cryptosporidiosis in these animals. This poses a risk to the trading of livestock, to other farms via breeding centres, and to human health. This study is a part of a global project aimed at strategies to tackle cryptosporidiosis. To reach this target, it was essential to determine whether prevalence was dependent on the studied countries or if the issue was borderless. Indeed, C. parvum occurrence was assessed across dairy farms in certain regions of Belgium, France, and the Netherlands. At the same time, the animal-to-animal transmission of the circulating C. parvum subtypes was studied. To accomplish this, we analysed 1084 faecal samples, corresponding to 57 dairy farms from all three countries. To this end, 18S rRNA and gp60 genes fragments were amplified, followed by DNA sequencing, which was subsequently used for detection and subtyping C. parvum. Bioinformatic and phylogenetic methods were integrated to analyse and characterise the obtained DNA sequences. Our results show 25.7%, 24.9% and 20.8% prevalence of Cryptosporidium spp. in Belgium, France, and the Netherlands respectively. Overall, 93% of the farms were Cryptosporidium positive. The gp60 subtyping demonstrated a significant number of the C. parvum positives belonged to the IIa allelic family, which has been also identified in humans. Therefore, this study highlights how prevalent C. parvum is in dairy farms and further suggests cattle as a possible carrier of zoonotic C. parvum subtypes, which could pose a threat to human health.

High Occurrence of Zoonotic Subtypes of Cryptosporidiumparvum in Cypriot Dairy Farms

“…Herein, the most numerically abundant C. parvum subtype was IIaA14G1R1 (red colour, Figure 2), and it was also the most widespread, having been found in 6/10 farms. This subtype has been reported previously in cows from Turkey [31], Austria [53,54], Estonia [55], Poland [56], Germany [57] and the Netherlands [24], though it was typically less abundant in all cases. Outbreaks of human cryptosporidiosis in Norway and New Zealand [58,59] have been attributed to IIaA14G1R1, and the subtype has also been found in human samples from the USA [60], Ethiopia [61], Slovenia [62] and Slovakia [63].…”

“…Samples were screened for Cryptosporidium spp. using nested-PCR amplification of the 631 bp region of the 18S rRNA gene [ 24 , 28 ]. Positive (genomic DNA from a pure culture of C. parvum ) and negative (water was used as template instead of DNA) controls were included in both the reactions.…”

“…Prevalence investigations have frequently been carried out in almost all continents [ 22 , 23 ]. Studies of Cryptosporidium occurrences in Europe point towards a high prevalence [ 24 , 25 , 26 ]. These investigations have primarily focused on mainland Europe, while island nations have been largely overlooked—with the exception of the UK [ 27 ].…”

High Occurrence of Zoonotic Subtypes of <em>Cryptosporidium parvum</em> in Cypriot Dairy Farms

“…Herein, the most numerically abundant C. parvum subtype was IIaA14G1R1 (red colour, Figure 2) and it was also the most widespread, having been found in 6/10 farms. This subtype has been reported previously in cows from Turkey [31], Austria [50,51], Estonia [52], Poland [53], Germany [54] and the Netherlands [24], though it was typically less abundant in all cases.…”

“…The subtype has also been identified in humans with diarrhoea from England [67], Australia [68], Egypt [46], Slovenia [59] and Lebanon [69]. IIaA18G2R1 has been previously identified in cattle in the USA [70], Northern Ireland [71], Germany [72], Italy [73] and France [24], though typically was one of the less common subtypes found. This is a potentially zoonotic subtype having been identified sporadically in humans from England [67], USA [57,74] and Australia [75].…”