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Cryptosporidiosis of calves is caused by the enteroprotozoan Cryptosporidium spp. The disease results in intense diarrhea of calves associated with substantial economic losses in dairy farming worldwide. The aim of this study was to determine calf, herd, and within-herd Cryptosporidium prevalence and identify Cryptosporidium species and subtypes in calves with diarrhea in intensive dairy herds in central Argentina. A total of 1073 fecal samples were collected from 54 randomly selected dairy herds. Cryptosporidium-oocysts were isolated and concentrated from fecal samples using formol-ether and detected by light microscopy with the modified Ziehl-Neelsen technique. Overall prevalence of oocyst-excreting calves was found to be 25.5% (274/1073) (95% C.I. 22.9; 28.1%). Of the herds studied, 89% (48/54) included at least one infected calf, whereas within-herd prevalence ranged from the absence of infection to 57% (20/35). A highly significant association was found between the presence of diarrhea and C. parvum infection (χ2 = 55.89, p < 0.001). For species determination, genomic DNA isolated from oocyst-positive fecal samples was subjected to PCR-RFLP of the 18S rRNA gene resulting exclusively in Cryptosporidium parvum identification. C. parvum isolates of calves displaying diarrhea and high rate of excretion of oocysts were subtyped by PCR amplification and direct sequencing of the 60 kDa glycoprotein (GP60) gene. Altogether five GP60 subtypes, designated IIaA18G1R1, IIaA20G1R1, IIaA21G1R1, IIaA22G1R1, and IIaA24G1R1 were identified. Interestingly, IIaA18G1R1 and IIaA20G1R1 were predominant in calves with diarrhea and high infection intensity. Notably, IIaA24G1R1 represents a novel, previously unrecognized C. parvum subtype. The subtype IIaA18G1R1, frequently found in this study, is strongly implicated in zoonotic transmission. These results suggest that calves might be an important source for human cryptosporidiosis in Argentina.

Background Cryptosporidium spp. are worldwide protozoan parasites which include species that can lead to cryptosporidiosis in humans. Different animal species can serve as reservoirs and sources of dissemination of the disease, such as rodent species due their potential in transmitting zoonotic pathogens to humans and other animals. In the Canary Islands (Spain), Cryptosporidium parvum and Cryptosporidium hominis have been identified in patients with diarrhea. However, the occurrence of Cryptosporidium spp. in possible reservoirs in this archipelago remains unclear. Considering the zoonotic potential of these protozoans, the aim of the present study was to determine the presence of Cryptosporidium spp. in peridomestic wild rodents and the possible role of these mammals as a source of transmission of these protozoans in Canary Islands. Methods A total of 179 rodents belonging to Rattus rattus and Mus musculus domesticus from four Canary Islands, La Palma, El Hierro, Tenerife and Lanzarote, were analyzed. Feces were screened for Cryptosporidium spp. by nested PCR of the 18S ribosomal RNA fragment and the sequences used for phylogenetic analyses. Results Cryptosporidium spp. were found widely distributed with an overall prevalence of 12.30% in rodents (13.86% for R. rattus and 10.25% for M. m. domesticus ). The overall prevalence by island was 19.60% for Tenerife, 7.14% for La Palma, 5.71% for El Hierro and 0% for Lanzarote. Cryptosporidium tyzzeri , Cryptosporidium meleagridis , Cryptosporidium muris and Cryptosporidium sp. rat genotype I and II/III were successfully identified, in addition to two unidentified Cryptosporidium genotypes. Conclusions This study contributes to the knowledge of the biodiversity and distribution of Cryptosporidium spp. in wild rodents from the Canary Islands, highlighting the presence of three zoonotic species, C. tyzzeri , C. meleagridis and C. muris , being the first detection of these three species in wild rodents in the Canary Islands and the first report of C. meleagridis in R. rattus . Given the results obtained in our study, future studies in non-sampled areas are required to better understand the epidemiology of these protozoans in wild rodents in the archipelago.

Background Little is known on the occurrence and identity of Cryptosporidium species in sheep and goats in Algeria. This study aimed at investigating the occurrence of Cryptosporidium species in lambs and goat kids younger than 4 weeks. Methods A total of 154 fecal samples (62 from lambs and 92 from kid goats) were collected from 13 sheep flocks in Médea, Algeria and 18 goat flocks across Algiers and Boumerdes. They were screened for Cryptosporidium spp. by nested-PCR analysis of a fragment of the small subunit ( SSU ) rRNA gene, followed by restriction fragment length polymorphism and sequence analyses to determine the Cryptosporidium species present. Cryptosporidium parvum and C. ubiquitum were further subtyped by sequence analysis of the 60 kDa glycoprotein gene. Results Cryptosporidium spp. were detected in 17 fecal samples (11.0%): 9 from lambs (14.5%) and 8 from goat kids (8.7%). The species identified included C. parvum in 3 lambs, C. xiaoi in 6 lambs and 6 goat kids, and C. ubiquitum in 2 goat kids. Cryptosporidium infections were detected mostly in animals during the first two weeks of life (7/8 for goat kids and 7/9 for lambs) and in association with diarrhea occurrence (7/17 or 41.2% goat kids and 7/10 or 70.0% lambs with diarrhea were positive for Cryptosporidium spp.). Subtyping of C. parvum and C. ubiquitum isolates identified the zoonotic IIaA13G2R1 and XIIa subtype families, respectively. Minor differences in the SSU rRNA gene sequences were observed between C. xiaoi from sheep and goats. Conclusions Results of this study indicate that three Cryptosporidium species occur in lambs and goat kids in Algeria, including zoonotic C. parvum and C. ubiquitum . They are associated with the occurrence of neonatal diarrhea.

PCR-based diagnostics has revealed the previously largely unknown transmission and infections in high-income countries. This study aimed to determine domestic and imported subtypes of species in Norway, evaluate their demographic distribution, and identify potential small outbreaks. -positive human faecal samples were obtained from six medical microbiology laboratories between February 2022 and January 2024, together with 22 -positive animal samples. Species and subtypes were identified by sequencing PCR products from gp60 and SSU rRNA genes. Most cryptosporidiosis cases occurred during late summer/early autumn, primarily in children and young adults. Of 550 human samples, 359 were successfully characterized molecularly (65%), revealing infection with 10 different species. occurred in 245 (68%) human isolates with IIa and IId being major allele families, with distinct regional distribution patterns of common subtypes. A kindergarten outbreak with 5 cases was due to IIaA14G1R1. was identified in 33 (9.2%) human cases of which 24 were known to be of domestic origin, making it the second most common species in human autochthonous cases in Norway. All isolates were of the same genotype; XIVaA20G2T1, including 13 cases from a suspected small outbreak in Trøndelag. occurred in 68 typed cases (19%), but mostly in infections acquired abroad, with allele families Ib and If occurring most often. In conclusion, this study of recent spp. and subtypes in Norway, highlights the predominance of and the emergence of among autochthonous cases.

Objective This study aimed to assess the prevalence of Cryptosporidium genotypes in the canal water bodies of Minya Al-Qamh District in Sharqia Governorate, Northern Egypt. Rural populations in Egypt lack access to clean water. They obtain their water supplies from different drains and canals, which are frequently exposed to contamination by human activities and untreated agricultural waste, introducing Cryptosporidium infection to unprotected waterways. A total of (72) canal water samples served for the molecular detection of Cryptosporidium species by PCR amplification and sequencing. Results Only one sample 1.4% (1/72) belonging to the village of Al-Aziziyyah was PCR-positive for Cryptosporidium contamination. Based on the ( COWP ) gene sequencing, this species was identified as Cryptosporidium parvum (C. parvum). The current work’s findings marked the first report on the molecular characterization of Cryptosporidium species in the canal water of Sharqia Province. Our results suggested that the source of C. parvum contamination could be of human and/or animal origin. Therefore, further studies are warranted to track the source of human infection and mitigate contamination for improved water quality.

Cryptosporidium species can infect humans and more than 260 animal species, including 54 rodent species. However, data on the occurrence and genetic characterizations of Cryptosporidium spp. in laboratory rodents are limited. The present study aimed to determine the occurrence rate and genetic characterizations of Cryptosporidium spp. in laboratory mice and rats. We collected 506 fresh combined fecal pellet specimens (457 from mice and 49 from rats) of more than 2,000 laboratory rodents in Heilongjiang Province and Shanghai City, China. Cryptosporidium spp. were identified and subtyped by DNA sequencing of the SSU rRNA and the gp60 genes, respectively. By sequence analysis of the SSU rRNA gene, the occurrence rate of Cryptosporidium spp. was 16.6% (84/506) in combined fecal specimens, with 18.2% (83/457) for mice and 2.0% (1/49) for rats. Cryptosporidium parvum ( n  = 39), C. tyzzeri ( n  = 33), and C. parvum  +  C. tyzzeri ( n  = 11) were identified in mice. Cryptosporidium parvum was only detected in one rat fecal specimen. At the gp60 locus, 71.4% (60/84) of the Cryptosporidium -positive specimens were successfully amplified, and they all came from mice. We identified five C. parvum subtypes (IIaA14G2R1, IIaA16G2R1, IIaA17G1R1, IIaA17G2R1, and IIaA18G2R1) and two C. tyzzeri subtypes (IXaA6R1 and IXbA8). Based on the identification in laboratory mice of C. parvum subtypes that have been reported previously in humans, the mice infected with this species may threaten human health, especially for people who have contact with the animals and their feces. Les espèces de Cryptosporidium peuvent infecter les humains et plus de 260 espèces animales, dont 54 espèces de rongeurs. Cependant, les données sur la présence et les caractérisations génétiques de Cryptosporidium spp. chez les rongeurs de laboratoire sont limitées. La présente étude visait à déterminer le taux de présence et les caractérisations génétiques de Cryptosporidium spp. chez des souris et des rats de laboratoire. Nous avons collecté 506 échantillons de boulettes fécales fraîches combinées (457 de souris et 49 de rats) de plus de 2 000 rongeurs de laboratoire dans la province du Heilongjiang et la ville de Shanghai, en Chine. Les Cryptosporidium spp. ont été identifiés et sous-typés par séquençage de l’ADN des gènes SSU rRNA et gp60 , respectivement. Par analyse de séquence du gène SSU rRNA, le taux de présence de Cryptosporidium spp. était de 16,6% (84/506) dans les échantillons fécaux combinés, avec 18,2 % (83/457) pour les souris et 2,0 % (1/49) pour les rats. Cryptosporidium parvum ( n  = 39), C. tyzzeri ( n  = 33) et C. parvum  +  C. tyzzeri ( n  = 11) ont été identifiés chez la souris. Cryptosporidium parvum n’a été détecté que dans un échantillon fécal de rat. Au locus gp60 , 71,4 % (60/84) des échantillons positifs à Cryptosporidium ont été amplifiés avec succès, et ils provenaient tous de souris. Nous avons identifié cinq sous-types de C. parvum (IIaA14G2R1, IIaA16G2R1, IIaA17G1R1, IIaA17G2R1 et IIaA18G2R1) et deux sous-types de C. tyzzeri (IXaA6R1 et IXbA8). Sur la base de l’identification, chez des souris de laboratoire, de sous-types de C. parvum qui ont déjà été signalés chez l’homme, les souris infectées par cette espèce peuvent menacer la santé humaine, en particulier pour les personnes qui sont en contact avec les animaux et leurs excréments.

Cryptosporidium is one of the most important parasitic protozoa of concern within the food production industry, worldwide. This review describes the evolution and its development, and it monitors the methodology that has been used for Cryptosporidium in food material since 1984, when the first publication appeared regarding the detection of Cryptosporidium parvum in food materials. The methods that are currently being used for the detection of Cryptosporidium oocysts in food material (mainly vegetables) and all of the other available published methods are discussed in this review. Generating more consistent and reliable data should lead to a better understanding of the occurrence, transport and fate of the oocysts in food material. Improvements in monitoring and developing effective methodology, along with food security, offer more practical possibilities for both the developed and developing worlds.

This study was conducted to molecularly identify and classify spp. in fecal samples (n=150) from patients with diarrhea received at the microbiology laboratory of a private hospital in Denizli. In this study, the positivity of spp. in fecal samples was investigated using direct microscopy, Kinyoun's acid-fast staining method, and Nested polymerase chain reaction (PCR) techniques. Positive PCR products were sequenced. In the examined fecal samples of patients with diarrhea, no parasites were detected through direct microscopic examination. Using the Kinyoun acid-fast staining method, spp. was identified in 2.7% (n=4) of the samples, while Nested PCR detected it in 4.67% (n=7) of the samples. The four positive samples were sequenced using primers that amplify the 18S gene region. The sequencing results identified the isolates as . Cryptosporidiosis is an important public health issue as it is a zoonotic disease caused by the parasite that can be transmitted from animals to humans. This study focuses on the molecular characterization of species detected in human fecal samples, which is significant for understanding which specific strains or species are involved in human infections. According to the findings, it is recommended that control measures be implemented to reduce the risk of exposure to in both humans and animals in Türkiye.

Background Whole genome sequencing (WGS) of Cryptosporidium spp. has previously relied on propagation of the parasite in animals to generate enough oocysts from which to extract DNA of sufficient quantity and purity for analysis. We have developed and validated a method for preparation of genomic Cryptosporidium DNA suitable for WGS directly from human stool samples and used it to generate 10 high-quality whole Cryptosporidium genome assemblies. Our method uses a combination of salt flotation, immunomagnetic separation (IMS), and surface sterilisation of oocysts prior to DNA extraction, with subsequent use of the transposome-based Nextera XT kit to generate libraries for sequencing on Illumina platforms. IMS was found to be superior to caesium chloride density centrifugation for purification of oocysts from small volume stool samples and for reducing levels of contaminant DNA. Results The IMS-based method was used initially to sequence whole genomes of Cryptosporidium hominis gp60 subtype IbA10G2 and Cryptosporidium parvum gp60 subtype IIaA19G1R2 from small amounts of stool left over from diagnostic testing of clinical cases of cryptosporidiosis. The C. parvum isolate was sequenced to a mean depth of 51.8X with reads covering 100 % of the bases of the C. parvum Iowa II reference genome (Bioproject PRJNA 15586), while the C. hominis isolate was sequenced to a mean depth of 34.7X with reads covering 98 % of the bases of the C. hominis TU502 v1 reference genome (Bioproject PRJNA 15585). The method was then applied to a further 17 stools, successfully generating another eight new whole genome sequences, of which two were C. hominis ( gp60 subtypes IbA10G2 and IaA14R3) and six C. parvum ( gp60 subtypes IIaA15G2R1 from three samples, and one each of IIaA17G1R1, IIaA18G2R1, and IIdA22G1), demonstrating the utility of this method to sequence Cryptosporidium genomes directly from clinical samples. This development is especially important as it reduces the requirement to propagate Cryptosporidium oocysts in animal models prior to genome sequencing. Conclusion This represents the first report of high-quality whole genome sequencing of Cryptosporidium isolates prepared directly from human stool samples.

Surveillance monitoring for microbial water quality typically involves collecting single discrete grab samples for analyzing only one contaminant. While informative, current approaches suffer from poor recoveries and only provide a limited snapshot of the microbial contaminants only at the time of collection. To overcome these limitations, bivalves have been proposed as effective biosentinels of water quality particularly for their ability to efficiently concentrate and retain microbial contaminants for long periods of time. In this study, we examined the use of indigenous blue mussels (Mytilus spp.) as biosentinels to monitor for the presence of Toxoplasma gondii and Cryptosporidium water. An efficient method to extract oocyst DNA from various mussel tissues followed by PCR-based detection of these pathogens was developed, which resulted in the detection down to 10 oocysts. This method was then used to conduct a small survey in Point Lobos and Morro Bay, California to determine prevalence T. gondii and Cryptosporidium. Results revealed that mussels from Morro Bay were contaminated with T. gondii (33 %), while mussels from Point Lobos were contaminated with T. gondii (54 %) and Cryptosporidium (26.9 %) oocysts. Phylogenetic analysis using the SSU rRNA gene identified two novel Cryptosporidium parvum-like genotypes. Overall, this study demonstrated the application of using native California Mytilus spp. as biosentinels for pathogen contamination along the central California shorelines. More importantly, T. gondii and Cryptosporidium were found at higher prevalence rates in Morro Bay and in Point Lobos, an area not previously reported to be contaminated with these pathogens.