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.

(1) Background: Blastocystis is a microbial eukaryote inhabiting the gastrointestinal tract of a broad range of animals including humans. Several studies have shown that the organism is associated with specific microbial profiles and bacterial taxa that have been deemed beneficial to intestinal and overall health. Nonetheless, these studies are focused almost exclusively on humans, while there is no similar information on other animals. (2) Methods: Using a combination of conventional PCR, cloning and sequencing, we investigated presence of Blastocystis along with Giardia and Cryptosporidium in 16 captive water voles sampled twice from a wildlife park. We also characterised their bacterial gut communities. (3) Results: Overall, alpha and beta diversities between water voles with and without Blastocystis did not differ significantly. Differences were noted only on individual taxa with Treponema and Kineothrix being significantly reduced in Blastocystis positive water voles. Grouping according to antiprotozoal treatment and presence of other protists did not reveal any differences in the bacterial community composition either. (4) Conclusion: Unlike human investigations, Blastocystis does not seem to be associated with specific gut microbial profiles in water voles.

Cryptosporidium parvum is one of the major causes of neonatal calf diarrhoea resulting in reduced farm productivity and compromised animal welfare worldwide. Livestock act as a major reservoir of this parasite, which can be transmitted to humans directly and/or indirectly, posing a public health risk. Research reports on the prevalence of Cryptosporidium in ruminants from east Mediterranean countries, including Cyprus, are limited. This study is the first to explore the occurrence of Cryptosporidium spp. in cattle up to 24 months old on the island of Cyprus. A total of 242 faecal samples were collected from 10 dairy cattle farms in Cyprus, all of which were screened for Cryptosporidium spp. using nested-PCR amplification targeting the small subunit of the ribosomal RNA (18S rRNA) gene. The 60 kDa glycoprotein (gp60) gene was also sequenced for the samples identified as Cryptosporidium parvum-positive to determine the subtypes present. The occurrence of Cryptosporidium was 43.8% (106/242) with at least one positive isolate in each farm sampled. Cryptosporidium bovis, Cryptosporidium ryanae and C. parvum were the only species identified, while the prevalence per farm ranged from 20­‒64%. Amongst these, the latter was the predominant species, representing 51.8% of all positive samples, followed by C. bovis (21.7%) and C. ryanae (31.1%). Five C. parvum subtypes were identified, four of which are zoonotic—IIaA14G1R1, IIaA15G1R1, IIaA15G2R1 and IIaA18G2R1. IIaA14G1R1 was the most abundant, representing 48.2% of all C. parvum positive samples, and was also the most widespread. This is the first report of zoonotic subtypes of C. parvum circulating in Cyprus. These results highlight the need for further research into the parasite focusing on its diversity, prevalence, host range and transmission dynamics on the island.

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Cryptosporidium parvum is one of the major causes of neonatal calf diarrhoea resulting in reduced farm productivity and compromised animal welfare worldwide. Livestock act as a major reservoir of this parasite, which can be transmitted to humans directly and/or indirectly, posing a public health risk. Research reports on Cryptosporidium prevalence from the east Mediterranean region are scarce with even less originating from Cyprus. This study is the first to explore occurrence of Cryptosporidium spp. in young dairy calves on the island. A total of 242 faecal samples were collected from 10 dairy cattle farms in Cyprus, all of which were screened for Cryptosporidium spp. using nested-PCR amplification targeting the small subunit of the ribosomal RNA (18S rRNA) gene. The 60 kDa glycoprotein (gp60) gene was also sequenced for the samples identified as C. parvum positive to determine the subtypes present. Occurrence of Cryptosporidium was 43.8% (106/242) with at least one positive isolate in each farm sampled. Prevalence per farm ranged from 20¬‒64%, while C. bovis, C. ryanae and C. parvum were the only species identified. Amongst those, the latter was the predominant species, representing 51.8% of all positive samples. Four zoonotic subtypes were identified – IIaA14G1R1, IIaA15G1R1, IIaA15G2R1 and IIaA18G2R1. IIaA14G1R1 was the most abundant; representing 48.2% of all C. parvum positive samples and was also the most widespread. This is the first report of zoonotic subtypes of C. parvum circulating in Cyprus. These results highlight the need for further research into the parasite focusing on its diversity, prevalence, host-range and transmission dynamics on the island.