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The diversity and biology of Cryptosporidium that is specific for rats (Rattus spp.) are not well studied. We examined the occurrence and genetic diversity of Cryptosporidium spp. in wild brown rats (Rattus norvegicus) by microscopy and polymerase chain reaction (PCR)/sequencing targeting the small subunit rDNA (SSU), actin and HSP70 genes. Out of 343 faecal samples tested, none were positive by microscopy and 55 were positive by PCR. Sequence analysis of SSU gene revealed the presence of Cryptosporidium muris (n = 4), C. andersoni (n = 3), C. ryanae (n = 1), C. occultus (n = 3), Cryptosporidium rat genotype I (n = 23), Cryptosporidium rat genotype IV (n = 16) and novel Cryptosporidium rat genotype V (n = 5). Spherical oocysts of Cryptosporidium rat genotype I obtained from naturally-infected rats, measuring 4.4–5.4 μm × 4.3–5.1 μm, were infectious to the laboratory rats, but not to the BALB/c mice (Mus musculus) nor Mongolian gerbils (Meriones unguiculatus). The prepatent period was 3 days post infection and the patent period was longer than 30 days. Naturally- and experimentally-infected rats showed no clinical signs of disease. Percentage of nucleotide similarities at the SSU, actin, HSP70 loci between C. ratti n. sp. and the rat derived C. occultus and Cryptosporidium rat genotype II, III, IV, and V ranged from 91.0 to 98.1%. These genetic variations were similar or greater than that observed between closely related species, i.e. C. parvum and C. erinacei (93.2–99.5%). Our morphological, genetic and biological data support the establishment of Cryptosporidium rat genotype I as a new species, Cryptosporidium ratti n. sp.

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 Cryptosporidiosis is a gastrointestinal disease with global distribution. It has been a reportable disease in Canada since 2000; however, routine molecular surveillance is not conducted. Therefore, sources of contamination are unknown. The aim of this project was to identify species and subtypes of Cryptosporidium in clinical cases from Ontario, the largest province in Canada, representing one third of the Canadian population, in order to understand transmission patterns. Methods A total of 169 frozen, banked, unpreserved stool specimens that were microscopy positive for Cryptosporidium over the period 2008–2017 were characterized using molecular tools. A subset of the 169 specimens were replicate samples from individual cases. DNA was extracted directly from the stool and nested PCR followed by Sanger sequencing was conducted targeting the small subunit ribosomal RNA (SSU) and glycoprotein 60 ( gp60 ) genes. Results Molecular typing data and limited demographic data were obtained for 129 cases of cryptosporidiosis. Of these cases, 91 (70.5 %) were due to Cryptosporidium parvum and 24 (18.6%) were due to Cryptosporidium hominis . Mixed infections of C. parvum and C. hominis occurred in four (3.1%) cases. Five other species observed were Cryptosporidium ubiquitum ( n = 5), Cryptosporidium felis ( n = 2), Cryptosporidium meleagridis ( n = 1), Cryptosporidium cuniculus ( n = 1) and Cryptosporidium muris ( n = 1). Subtyping the gp60 gene revealed 5 allelic families and 17 subtypes of C. hominis and 3 allelic families and 17 subtypes of C. parvum . The most frequent subtype of C. hominis was IbA10G2 (22.3%) and of C. parvum was IIaA15G2R1 (62.4%). Conclusions The majority of isolates in this study were C. parvum , supporting the notion that zoonotic transmission is the main route of cryptosporidiosis transmission in Ontario. Nonetheless, the observation of C. hominis in about a quarter of cases suggests that anthroponotic transmission is also an important contributor to cryptosporidiosis pathogenesis in Ontario. Graphical Abstract

The morphological, biological, and molecular characteristics of Cryptosporidium muris strain TS03 are described, and the species name Cryptosporidium proliferans n. sp. is proposed. Cryptosporidium proliferans obtained from a naturally infected East African mole rat (Tachyoryctes splendens) in Kenya was propagated under laboratory conditions in rodents (SCID mice and southern multimammate mice, Mastomys coucha) and used in experiments to examine oocyst morphology and transmission. DNA from the propagated C. proliferans isolate, and C. proliferans DNA isolated from the feces of an African buffalo (Syncerus caffer) in Central African Republic, a donkey (Equus africanus) in Algeria, and a domestic horse (Equus caballus) in the Czech Republic were used for phylogenetic analyses. Oocysts of C. proliferans are morphologically distinguishable from C. parvum and C. muris HZ206, measuring 6.8-8.8 (mean = 7.7 μm) × 4.8-6.2 μm (mean = 5.3) with a length to width ratio of 1.48 (n = 100). Experimental studies using an isolate originated from T. splendens have shown that the course of C. proliferans infection in rodent hosts differs from that of C. muris and C. andersoni. The prepatent period of 18-21 days post infection (DPI) for C. proliferans in southern multimammate mice (Mastomys coucha) was similar to that of C. andersoni and longer than the 6-8 DPI prepatent period for C. muris RN66 and HZ206 in the same host. Histopatologicaly, stomach glands of southern multimammate mice infected with C. proliferans were markedly dilated and filled with necrotic material, mucus, and numerous Cryptosporidium developmental stages. Epithelial cells of infected glands were atrophic, exhibited cuboidal or squamous metaplasia, and significantly proliferated into the lumen of the stomach, forming papillary structures. The epithelial height and stomach weight were six-fold greater than in non-infected controls. Phylogenetic analyses based on small subunit rRNA, Cryptosporidium oocyst wall protein, thrombospondin-related adhesive protein of Cryptosporidium-1, heat shock protein 70, actin, heat shock protein 90 (MS2), MS1, MS3, and M16 gene sequences revealed that C. proliferans is genetically distinct from C. muris and other previously described Cryptosporidium species.

Background Cryptosporidium infection occurs in humans, domestic animals, and wildlife. To date, at least 49 species and 120 genotypes have been identified. Hitherto, molecular identification of Cryptosporidium species in wildlife has seldom been reported in China. Methods During 2014–2025, a total of 1855 small intestinal or fecal specimens were collected from 1500 mammals, 121 reptiles, and 234 birds in Xinjiang Uygur Autonomous Region (XUAR) and Inner Mongolia Autonomous Region (IMAR), northern China. The identification of each animal species was based on morphological characteristics and mitochondrial gene amplification. Detection of  Cryptosporidium  species was performed by amplifying part of the small subunit ( SSU ) ribosomal RNA ( rRNA ) gene. The 60 kDa glycoprotein ( GP60 ) gene was used to confirm their species and subtypes. Results The samples were collected from 39 mammalian, 6 reptilian, and 30 avian species. In these samples, the average rate of infection with Cryptosporidium  species was 8.09% (150/1855). In total, 18 known Cryptosporidium species and genotypes were identified, including Cryptosporidium hominis , Cryptosporidium ubiquitum , Cryptosporidium muris , Cryptosporidium canis , Cryptosporidium felis , Cryptosporidium equi , Cryptosporidium proventriculi , Cryptosporidium ryanae , Cryptosporidium rubeyi , chipmunk genotype V, vole genotype III, vole genotype V, muskrat genotype I, bat genotype IV, yak genotype, deer genotype, goose genotype I, and one unnamed Cryptosporidium sp. In addition, a novel genotype, here designated as Cryptosporidium Mongolian pika genotype, was identified in the Mongolian pika ( Ochotona pallasi ). Conclusions Investigation of Cryptosporidium infection was carried out by screening 75 animal species. Overall, 19 Cryptosporidium species and genotypes were detected, including a novel genotype in Mongolian pika and first-time diagnosis of this infection in several rodent species (e.g., red-cheeked ground squirrels, great gerbils, northern mole voles, and Libyan jirds). Graphical Abstract

The North African hedgehog ( Atelerix algirus ) is an introduced species from Northwest Africa and is currently distributed in the Canary Islands. This species of hedgehog has been studied as a reservoir of enteropathogens, including Cryptosporidium spp. However, there are no data at species level. Therefore, the aim of the present study was to identify the Cryptosporidium species present in a population of hedgehogs ( n  = 36) in the Canary Islands. Molecular screening was performed using conventional polymerase chain reaction (PCR) targeting the small subunit ribosomal RNA ( 18S rRNA ) gene of Cryptosporidium spp. Seven of the 36 fecal samples (19.45%) were positive and confirmed by nested PCR targeting the 18S rRNA gene and Sanger sequencing. Cryptosporidium parvum and Cryptosporidium muris were identified in 11.1% (4/36) and 5.6% (2/36) of the samples, respectively, while one sample could only be identified at the genus level. The zoonotic subtypes IIdA15G1 ( n  = 1), IIdA16G1b ( n  = 1), and IIdA22G1 ( n  = 1) of C. parvum were identified by nested PCR followed by analysis of the 60 kDa glycoprotein ( gp60 ) gene sequence. This study is the first genetic characterization of Cryptosporidium spp. in A. algirus , identifying zoonotic species and subtypes of the parasite.

Cryptosporidium and Giardia are two important zoonotic intestinal parasites responsible for diarrhoea in humans and other animals worldwide. Rodents, as reservoirs or carriers of Cryptosporidium and Giardia, are abundant and globally widespread. In the present study, we collected 232 fecal specimens from commensal rodents captured in animal farms and farm neighbourhoods in China. We collected 33 Asian house rats, 168 brown rats and 31 house mice. 6·0% (14/232) and 8·2% (19/232) of these rodents were microscopy-positive for Giardia cysts and Cryptosporidium oocysts, respectively. All 14 Giardia isolates were identified as Giardia duodenalis assemblage G at a minimum of one or maximum of three gene loci (tpi, gdh and bg). By small subunit rRNA (SSU rRNA) gene sequencing, Cryptosporidium parvum (n = 12) and Cryptosporidium muris (n = 7) were identified. The gp60 gene encoding the 60-kDa glycoprotein was successfully amplified and sequenced in nine C. parvum isolates, all of which belonged to the IIdA15G1 subtype. Observation of the same IIdA15G1 subtype in humans (previously) and in rodents (here) suggests that rodents infected with Cryptosporidium have the potential to transmit cryptosporidiosis to humans.

Giardia duodenalis is a common and widespread intestinal protozoan parasite of both humans and animals. Previous epidemiological and molecular studies have identified Giardia infections in different animals and humans, but only limited information is available about the occurrence and genotypes of Giardia in cattle in China. In this study, we determined the occurrence of giardiasis and genetically characterized G. duodenalis in dairy cattle in Henan Province, central China. The overall prevalence of G. duodenalis was 7.2% (128/1777) on microscopic analysis, with the highest infection rate (22.7%) in calves aged less than 1 month. G. duodenalis assemblages and subtypes were identified with multilocus genotyping based on the SSU rRNA, β-giardin (bg), glutamate dehydrogenase (gdh), and triosephosphate isomerase (tpi) genes. Two assemblages were detected in the successfully sequenced samples: assemblage A (n = 58), assemblage E (n = 21), with a mixed E and A assemblage (n = 2). Four novel subtypes of the gdh gene and seven of the bg gene were found among the G. duodenalis assemblage E isolates. Using the nomenclature for the multilocus genotype (MLG) model, nine novel multilocus genotypes E (MLGs E1-E9) and three MLGs A (a novel subtype AI, previously detected subtype AII-1, and a combination of both) were identified. MLG AII-1 identified in this study may be an important zoonotic subtype. The dairy cattle in Henan are a potential public health concern.

Cryptosporidium is a significant zoonotic parasite with broad distribution in both humans and rodents. In this study, 510 fecal samples were collected from nine species of wild rodents across Guangxi, Yunnan, and Hunan Provinces in China. Nested PCR analysis targeting the SSU rRNA gene revealed an overall Cryptosporidium infection rate of 1.8% (9/510) among rodents in these provinces. The highest positivity rate was observed in Guangxi Province at 4.9% (5/103), followed by Yunnan Province (2.3%, 2/88), and Hunan Province (0.6%, 2/319). Notably, Rattus losea exhibited the highest prevalence rate at 9.8% (4/41), while Rattus flavipectus and Niviventer lotipes showed rates of 5.1% (2/39) and 4.4% (1/23), respectively. Various genotypes/species were identified, including Cryptosporidium viatorum , Cryptosporidium muris , Cryptosporidium vole genotype VII, and Cryptosporidium ratti , rat genotypes II, and IV. The study also found that wild rodents inhabiting mountainous areas had a higher prevalence rate at 4.9% (5/103) compared to those residing in fields and lake beaches, where prevalence rates were 2.1% (2/95) and 0.6% (2/312), respectively. This study provides new insights into Cryptosporidium infection rates among wild rodents and identifies two zoonotic species, C. viatorum and C. muris . These findings underscore the potential risk posed by Chinese wild rodent populations in transmitting zoonotic Cryptosporidium , which could significantly impact public health. Therefore, effective control strategies are needed to prevent transmission between humans and rodents. Cryptosporidium est un parasite zoonotique important avec une large distribution chez l’homme et les rongeurs. Dans cette étude, 510 échantillons fécaux ont été collectés de neuf espèces de rongeurs sauvages dans les provinces du Guangxi, du Yunnan et du Hunan en Chine. L’analyse par PCR imbriquée ciblant le gène ARNr SSU a révélé un taux global d’infection par Cryptosporidium de 1,8 % (9/510) parmi les rongeurs de ces provinces. Le taux de positivité le plus élevé a été observé dans la province du Guangxi avec 4,9 % (5/103), suivie de la province du Yunnan (2,3 %, 2/88) et de la province du Hunan (0,6 %, 2/319). Notamment, Rattus losea a présenté le taux de prévalence le plus élevé à 9,8 % (4/41), tandis que Rattus flavipectus et Niviventer lotipes ont affiché des taux de 5,1 % (2/39) et 4,4 % (1/23), respectivement. Divers génotypes/espèces ont été identifiés, notamment Cryptosporidium viatorum , Cryptosporidium muris , Cryptosporidium de campagnol génotype VII et Cryptosporidium ratti , génotypes de rats II et IV. L’étude a également révélé que les rongeurs sauvages habitant les zones montagneuses avaient un taux de prévalence plus élevé, de 4,9 % (5/103), par rapport à ceux résidant dans les champs et les bords de lacs, où les taux de prévalence étaient respectivement de 2,1 % (2/95) et de 0,6 % (2/312). Cette étude apporte de nouvelles perspectives sur les taux d’infection à Cryptosporidium chez les rongeurs sauvages et identifie deux espèces zoonotiques, C. viatorum et C. muris . Ces résultats soulignent le risque potentiel posé par les populations de rongeurs sauvages chinois dans la transmission des Cryptosporidium zoonotiques, ce qui pourrait avoir des conséquences importantes sur la santé publique. Par conséquent, des stratégies de contrôle efficaces sont nécessaires pour prévenir la transmission entre les humains et les rongeurs.

Background There are only limited number of reports on molecular epidemiology of Cryptosporidium spp. and Giardia duodenalis in dogs and cats in China. This study was conducted to assess the infection rates, genetic identity, and public health potential of these parasites in dogs and cats in Guangdong, China. Methods PCR and sequence analyses were used to identify and genotype Cryptosporidium spp. and G. duodenalis in fecal samples from 641 dogs and 418 cats in Guangdong. Chi-square test and odds ratio analysis were used to compare the occurrence rates of these pathogens and identify risk factors for infection. Results The overall infection rates of Cryptosporidium spp. and G. duodenalis were 6.9% (44/641) and 9.4% (60/641) in dogs, and 6.2% (26/418) and 3.6% (15/418) in cats. Purebred cats (12.4%; χ 2  = 5.110, OR = 2.8, P  = 0.024) and dogs (10.8%; χ 2  = 5.597, OR = 4.8, P  = 0.018) were more likely to be infected by Cryptosporidium spp. and G. duodenalis , respectively. Dogs (12.0%; χ 2  = 7.589, OR = 2.6, P  = 0.006) and cats (13.6%; χ 2  = 8.235, OR = 3.5, P  = 0.004) under 6 months had significantly higher infection rates of Cryptosporidium spp. than older animals. Household (13.9%; χ 2  = 10.279, OR = 2.6, P  = 0.008) and pet shop dogs (11.0%; χ 2  = 7.182, OR = 2.0, P  = 0.048) had higher occurrence of Cryptosporidium spp., as was the case for G. duodenalis occurrence in experimental dogs (13.4%; χ 2  = 9.223, OR = 1.9, P  = 0.017). Cryptosporidium canis ( n  = 42), C. muris ( n  = 1) and Cryptosporidium rat genotype IV ( n  = 1) were identified in dogs, while C. felis ( n  = 21), C. parvum ( n  = 3), C. muris ( n  = 1) and Cryptosporidium rat genotype IV ( n  = 1) were identified in cats. In contrast, the canine-specific assemblages C ( n  = 27) and D ( n  = 26) and the feline-specific assemblage F ( n  = 14) were almost exclusively the only genotypes of G. duodenalis in dogs and cats, respectively. There was no significant difference in infection rates of Cryptosporidium spp. and G . duodenalis between diarrheal and non-diarrheal pets. Conclusions While domestic pets in Guangdong are infected with zoonotic Cryptosporidium species, they are mainly infected with host-specific G. duodenalis genotypes. Risk factors for infections differ between Cryptosporidium spp. and G. duodenalis and between dogs and cats.