RESUMEN
Cryptosporidiosis has previously been reported in animals, humans, and water sources in the United Arab Emirates (UAE). However, most reports were only to the genus level, or generically identified as cryptosporidiosis. We aimed to investigate the genetic diversity of Cryptosporidium species occurring in diarrhetic ungulates which were brought to the Central Veterinary Research Laboratory (CVRL) in Dubai. Using a combination of microscopic and molecular methods, we identified five species of Cryptosporidium occurring among ungulates in the UAE, namely C. parvum, C. hominis, C. xiaoi, C. meleagridis, and C. equi. Cryptosporidium parvum was the most prevalent species in our samples. Furthermore, we identified subtypes of C. parvum and C. hominis, which are involved in both human and animal cryptosporidiosis. This is also the first reported occurrence of Cryptosporidium spp. in the Arabian Tahr, to our knowledge. Since the animals examined were all in contact with humans, the possibility of zoonotic spread is possible. Our study correlates with previous reports in the region, building upon the identification of Cryptosporidium sp. However, there is a need to further investigate the endemic populations of Cryptosporidium, including more hosts, sampling asymptomatic animals, and location data.
Asunto(s)
Criptosporidiosis , Cryptosporidium , Diarrea , Variación Genética , Emiratos Árabes Unidos/epidemiología , Animales , Criptosporidiosis/parasitología , Criptosporidiosis/epidemiología , Cryptosporidium/genética , Cryptosporidium/clasificación , Cryptosporidium/aislamiento & purificación , Diarrea/veterinaria , Diarrea/parasitología , Diarrea/epidemiología , Heces/parasitología , Bovinos , Filogenia , Cabras/parasitología , ADN Protozoario/genéticaRESUMEN
BACKGROUND: Calf diarrhea is a major cause of morbidity and mortality in the livestock sector worldwide and it can be caused by multiple infectious agents. In Ethiopia, cattle are the most economically important species within the livestock sector, but at the same time the young animals suffer from high rates of morbidity and mortality due to calf diarrhea. However, studies including both screening and molecular characterization of bovine enteric pathogens are lacking. Therefore, we aimed to both detect and molecularly characterize four of the major enteric pathogens in calf diarrhea, Enterotoxigenic Escherichia coli (E. coli K99 +), Cryptosporidium spp., rotavirus A (RVA), and bovine coronavirus (BCoV) in calves from central Ethiopia. Diarrheic and non-diarrheic calves were included in the study and fecal samples were analyzed with antigen-ELISA and quantitative real-time PCR (qPCR). Positive samples were further characterized by genotyping PCRs. RESULTS: All four pathogens were detected in both diarrheic and non-diarrheic calves using qPCR and further characterization showed the presence of three Cryptosporidium species, C. andersoni, C. bovis and C. ryanae. Furthermore, genotyping of RVA-positive samples found a common bovine genotype G10P[11], as well as a more unusual G-type, G24. To our knowledge this is the first detection of the G24 RVA genotype in Ethiopia as well as in Africa. Lastly, investigation of the spike gene revealed two distinct BCoV strains, one classical BCoV strain and one bovine-like CoV strain. CONCLUSIONS: Our results show that Cryptosporidium spp., E. coli K99 + , RVA and BCoV circulate in calves from central Ethiopia. Furthermore, our findings of the rare RVA G-type G24 and a bovine-like CoV demonstrates the importance of genetic characterization.
Asunto(s)
Enfermedades de los Bovinos , Coronavirus Bovino , Cryptosporidium , Diarrea , Heces , Rotavirus , Animales , Bovinos , Etiopía/epidemiología , Diarrea/veterinaria , Diarrea/virología , Diarrea/microbiología , Diarrea/parasitología , Enfermedades de los Bovinos/virología , Enfermedades de los Bovinos/epidemiología , Enfermedades de los Bovinos/microbiología , Enfermedades de los Bovinos/parasitología , Heces/virología , Heces/parasitología , Heces/microbiología , Rotavirus/genética , Rotavirus/aislamiento & purificación , Rotavirus/clasificación , Cryptosporidium/aislamiento & purificación , Cryptosporidium/genética , Cryptosporidium/clasificación , Coronavirus Bovino/genética , Coronavirus Bovino/aislamiento & purificación , Escherichia coli Enterotoxigénica/aislamiento & purificación , Escherichia coli Enterotoxigénica/genética , Genotipo , Criptosporidiosis/epidemiología , Infecciones por Rotavirus/veterinaria , Infecciones por Rotavirus/epidemiología , Infecciones por Rotavirus/virología , Infecciones por Coronavirus/veterinaria , Infecciones por Coronavirus/virología , Infecciones por Coronavirus/epidemiología , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Infecciones por Escherichia coli/veterinaria , Infecciones por Escherichia coli/epidemiología , Infecciones por Escherichia coli/microbiologíaRESUMEN
Snakes are sometimes regarded as pets and are used in traditional Chinese medicine. Cryptosporidium spp. are frequently identified in snakes, representing an important pathogen and causing gastrointestinal diseases. Current data indicate that risk factors for infection and patterns of clinical symptom presentation may differ among Cryptosporidium spp. To better understand the infection status by Cryptosporidium spp., fecal samples were collected from 603 asymptomatic and 147 symptomatic snakes in 26 provinces of China. These samples came from Elaphe guttata, Elaphe obsoleta, Pituophis melanoleucus, Thamnophis sirtalis, Lampropeltis getulus, and Heterodon nasicus. The partial small subunit (SSU) rRNA gene was amplified using nested polymerase chain reaction (PCR) to investigate the infection rate of Cryptosporidium spp., and to assess evolutionary relationships and genetic characterization. A prevalence of 20% was recorded in asymptomatic snakes, with age identified as a significant risk factor. In contrast, 70% of symptomatic snakes were positive for Cryptosporidium spp., with Cryptosporidium serpentis and Cryptosporidium varanii (syn. C. saurophilum). Further analysis revealed a potential association between C. serpentis and regurgitation, and C. varanii and diarrhea, while neither species was linked to flatulence. To our knowledge, this is the first study to report Cryptosporidium spp. and associated clinical signs in symptomatic snakes in China. This study aims to enhance the understanding of Cryptosporidium infections, risk factors, and clinical manifestations in snakes, providing data crucial for the control and prevention of cryptosporidiosis.
Title: Cryptosporidium spp. chez les serpents captifs de 26 provinces de Chine : prévalence, caractérisation moléculaire et symptômes. Abstract: Les serpents sont parfois considérés comme animaux de compagnie et sont utilisés en médecine traditionnelle chinoise. Des Cryptosporidium spp. sont fréquemment identifiés chez les serpents, ont un rôle d'agent pathogène important et provoquent des maladies gastro-intestinales. Les données actuelles indiquent que les facteurs de risque d'infection et les schémas de présentation des symptômes cliniques peuvent varier en fonction des espèces de Cryptosporidium. Pour mieux comprendre l'état d'infection par Cryptosporidium spp., des échantillons fécaux ont été collectés auprès de 603 serpents asymptomatiques et 147 serpents symptomatiques dans 26 provinces de Chine. Ces échantillons provenaient d'Elaphe guttata, Elaphe obsoleta, Pituophis melanoleucus, Thamnophis sirtalis, Lampropeltis getulus et Heterodon nasicus. Le gène de l'ARNr de la petite sous-unité partielle (SSU) a été amplifié à l'aide d'une réaction en chaîne par polymérase (PCR) imbriquée pour étudier le taux d'infection par Cryptosporidium spp. et évaluer les relations évolutives et la caractérisation génétique. Une prévalence de 20 % a été trouvée chez les serpents asymptomatiques, l'âge étant identifié comme un facteur de risque important. En revanche, 70 % des serpents symptomatiques étaient positifs à Cryptosporidium spp. avec Cryptosporidium serpentis et Cryptosporidium varanii (syn. C. saurophilum). Une analyse plus approfondie a révélé une association potentielle entre C. serpentis et la régurgitation, et C. varanii et la diarrhée, alors qu'aucune des deux espèces n'était liée aux flatulences. À notre connaissance, il s'agit ici de la première étude à signaler la présence de Cryptosporidium spp. et les signes cliniques associés chez des serpents symptomatiques en Chine. Cette étude vise à améliorer la compréhension des infections à Cryptosporidium, des facteurs de risque et des manifestations cliniques chez les serpents, en fournissant des données cruciales pour le contrôle et la prévention de la cryptosporidiose.
Asunto(s)
Criptosporidiosis , Cryptosporidium , Heces , Serpientes , Animales , Cryptosporidium/genética , Cryptosporidium/aislamiento & purificación , Cryptosporidium/clasificación , Criptosporidiosis/epidemiología , Criptosporidiosis/parasitología , China/epidemiología , Prevalencia , Heces/parasitología , Serpientes/parasitología , Filogenia , Factores de Riesgo , Reacción en Cadena de la Polimerasa/veterinaria , Masculino , Femenino , ADN Protozoario/aislamiento & purificación , Diarrea/parasitología , Diarrea/veterinaria , Diarrea/epidemiología , Mascotas/parasitologíaRESUMEN
The domestic chinchilla (Chinchilla lanigera) is kept as a pet and previous studies suggest that it may play an important role as a source of zoonotic parasites, including Giardia intestinalis, Cryptosporidium spp. and microsporidia. In this study, we examined the occurrence and genetic diversity of above mentioned parasites in pet chinchillas in the Czech Republic by PCR/sequencing of the 18S rRNA, TPI, and ITS genes. Of 149 chinchillas from 24 breeders, 91.3â¯% were positive for G. intestinalis, 8.1â¯% for Cryptosporidium spp., 2.0â¯% for Encephalitozoon spp., and 5.4â¯% for E. bieneusi. Molecular analyses revealed presence of G. intestinalis assemblage B, C. ubiquitum (XIIa family), E. bieneusi genotypes D, SCF2, and, CHN-F1, and E. intestinalis. The infection intensity of G. intestinalis determined by qRT-PCR reached up to 53,978 CPG, C. ubiquitum up to 1409 OPG, E. intestinalis up to 1124 SPG, and E. bieneusi up to 1373 SPG. Only two chinchillas with C. ubiquitum and five with G. intestinalis had diarrhoea at the time of the screening. Three chinchillas in the long-term study were consistently positive for G. intestinalis, with intermittent excretion of C. ubiquitum, E. intestinalis, and E. bieneusi over 25 weeks. The findings indicate that chinchillas are frequently infected with zoonotic parasitic protists, but that these infections rarely show clinical signs. The lack of visible signs could reduce the vigilance of pet owners when handling their chinchillas, increasing the risk of transmission within breeding groups and possibly to humans.
Asunto(s)
Chinchilla , Cryptosporidium , Encephalitozoon , Encefalitozoonosis , Enterocytozoon , Giardia lamblia , Giardiasis , Microsporidiosis , Mascotas , Zoonosis , Animales , Chinchilla/parasitología , Encephalitozoon/genética , Encephalitozoon/aislamiento & purificación , Encephalitozoon/clasificación , Zoonosis/parasitología , Cryptosporidium/genética , Cryptosporidium/clasificación , Cryptosporidium/aislamiento & purificación , Giardiasis/veterinaria , Giardiasis/parasitología , Giardia lamblia/genética , Giardia lamblia/aislamiento & purificación , Giardia lamblia/clasificación , República Checa/epidemiología , Encefalitozoonosis/veterinaria , Encefalitozoonosis/epidemiología , Encefalitozoonosis/microbiología , Enterocytozoon/genética , Enterocytozoon/aislamiento & purificación , Microsporidiosis/veterinaria , Microsporidiosis/epidemiología , Criptosporidiosis/parasitología , Criptosporidiosis/epidemiología , ARN Ribosómico 18S/genética , Heces/parasitología , Heces/microbiología , MasculinoRESUMEN
Freshwater samples (n = 199) were obtained from 41 sites with contrasting land-uses (avian, low impact, dairy, urban, sheep and beef, and mixed sheep, beef and dairy) and the E. coli phylotype of 3980 isolates (20 per water sample enrichment) was determined. Eight phylotypes were identified with B1 (48.04%), B2 (14.87%) and A (14.79%) the most abundant. Escherichia marmotae (n = 22), and Escherichia ruysiae (n = 1), were rare (0.68%) suggesting that these environmental strains are unlikely to confound water quality assessments. Phylotypes A and B1 were overrepresented in dairy and urban sites (p < 0.0001), whilst B2 were overrepresented in low impact sites (p < 0.0001). Pathogens ((Salmonella, Campylobacter, Cryptosporidium or Giardia) and the presence of diarrhoeagenic E. coli-associated genes (stx and eae) were detected in 89.9% (179/199) samples, including 80.5% (33/41) of samples with putative non-recent faecal inputs. Quantitative PCR to detect microbial source tracking targets from human, ruminant and avian contamination were concordant with land-use type and E. coli phylotype abundance. This study demonstrated that a potential recreational health risk remains where pathogens occurred in water samples with low E. coli concentration, potential non-recent faecal sources, low impact sites and where human, ruminant and avian faecal sources were absent.
Asunto(s)
Escherichia coli , Agua Dulce , Salud Pública , Calidad del Agua , Nueva Zelanda , Escherichia coli/genética , Escherichia coli/aislamiento & purificación , Escherichia coli/clasificación , Agua Dulce/microbiología , Animales , Humanos , Microbiología del Agua , Filogenia , Heces/microbiología , Cryptosporidium/genética , Cryptosporidium/aislamiento & purificación , Cryptosporidium/clasificación , Giardia/genética , Giardia/aislamiento & purificación , Giardia/clasificaciónRESUMEN
Wild rodents serve as reservoirs for Cryptosporidium and are overpopulated globally. However, genetic data regarding Cryptosporidium in these animals from China are limited. Here, we have determined the prevalence and genetic characteristics of Cryptosporidium among 370 wild rodents captured from three distinct locations in the southern region of Zhejiang Province, China. Fresh feces were collected from the rectum of each rodent, and DNA was extracted from them. The rodent species was identified by PCR amplifying the vertebrate cytochrome b gene. Cryptosporidium was detected by PCR amplification and amplicon sequencing the small subunit of ribosomal RNA gene. Positive samples of C. viatorum and C. parvum were further subtyped by analyzing the 60-kDa glycoprotein gene. A positive Cryptosporidium result was found in 7% (26/370) of samples, involving five rodent species: Apodemus agrarius (36), Niviventer niviventer (75), Rattus losea (18), R. norvegicus (155), and R. tanezumi (86). Their respective Cryptosporidium positive rates were 8.3%, 5.3%, 11.1%, 7.1%, and 7.0%. Sequence analysis confirmed the presence of three Cryptosporidium species: C. parvum (4), C. viatorum (1), and C. muris (1), and two genotypes: Cryptosporidium rat genotype IV (16) and C. mortiferum-like (4). Additionally, two subtypes of C. parvum (IIdA15G1 and IIpA19) and one subtype of C. viatorum (XVdA3) were detected. These results demonstrate that various wild rodent species in Zhejiang were concurrently infected with rodent-adapted and zoonotic species/genotypes of Cryptosporidium, indicating that these rodents can play a role in maintaining and dispersing this parasite into the environment and other hosts, including humans.
Title: Transmission interspécifique de Cryptosporidium chez les rongeurs sauvages de la région sud de la province chinoise du Zhejiang et son impact possible sur la santé publique. Abstract: Les rongeurs sauvages servent de réservoirs à Cryptosporidium et ont des grandes populations à l'échelle mondiale. Cependant, les données génétiques concernant Cryptosporidium chez ces animaux en Chine sont limitées. Ici, nous avons déterminé la prévalence et les caractéristiques génétiques de Cryptosporidium parmi 370 rongeurs sauvages capturés dans trois endroits distincts de la région sud de la province du Zhejiang, en Chine. Des excréments frais ont été collectés dans le rectum de chaque rongeur et l'ADN en a été extrait. L'espèce de rongeur a été identifiée par amplification par PCR du gène du cytochrome b des vertébrés. Cryptosporidium a été détecté par amplification PCR et séquençage d'amplicons de la petite sous-unité du gène de l'ARN ribosomal. Les échantillons positifs de C. viatorum et C. parvum ont ensuite été sous-typés en analysant le gène de la glycoprotéine de 60 kDa. Un résultat positif pour Cryptosporidium a été trouvé dans 7 % (26/370) des échantillons, impliquant cinq espèces de rongeurs : Apodemus agrarius (36), Niviventer niviventer (75), Rattus losea (18), R. norvegicus (155) et R. tanezumi (86). Leurs taux respectifs de positivité pour Cryptosporidium étaient de 8,3 %, 5,3 %, 11,1 %, 7,1 % et 7,0 %. L'analyse des séquences a confirmé la présence de trois espèces de Cryptosporidium : C. parvum (4), C. viatorum (1) et C. muris (1), et de deux génotypes : Cryptosporidium génotype IV de rat (16) et C. mortiferum-like (4). De plus, deux sous-types de C. parvum (IIdA15G1 et IIpA19) et un sous-type de C. viatorum (XVdA3) ont été détectés. Ces résultats démontrent que diverses espèces de rongeurs sauvages du Zhejiang sont simultanément infectées par des espèces/génotypes de Cryptosporidium zoonotiques et adaptés aux rongeurs, ce qui indique que ces rongeurs peuvent jouer un rôle dans le maintien et la dispersion de ce parasite dans l'environnement et d'autres hôtes, y compris les humains.
Asunto(s)
Animales Salvajes , Criptosporidiosis , Cryptosporidium , Heces , Enfermedades de los Roedores , Roedores , Animales , Criptosporidiosis/epidemiología , Criptosporidiosis/parasitología , Criptosporidiosis/transmisión , China/epidemiología , Cryptosporidium/genética , Cryptosporidium/aislamiento & purificación , Cryptosporidium/clasificación , Heces/parasitología , Enfermedades de los Roedores/parasitología , Enfermedades de los Roedores/epidemiología , Enfermedades de los Roedores/transmisión , Animales Salvajes/parasitología , Ratas/parasitología , Roedores/parasitología , Prevalencia , Salud Pública , Reservorios de Enfermedades/parasitología , Reservorios de Enfermedades/veterinaria , Filogenia , Humanos , ADN Protozoario/aislamiento & purificación , Murinae/parasitología , Reacción en Cadena de la Polimerasa , Zoonosis/parasitología , Zoonosis/transmisión , Zoonosis/epidemiología , GenotipoRESUMEN
Genetic variation in Cryptosporidium, a common protozoan gut parasite in humans, is often based on marker genes containing trinucleotide repeats, which differentiate subtypes and track outbreaks. However, repeat regions have high replication slippage rates, making it difficult to discern biological diversity from error. Here, we synthesized Cryptosporidium DNA in clonal plasmid vectors, amplified them in different mock community ratios, and sequenced them using next-generation sequencing to determine the rate of replication slippage with dada2. Our results indicate that slippage rates increase with the length of the repeat region and can contribute to error rates of up to 20%.
Asunto(s)
Cryptosporidium , Replicación del ADN , Cryptosporidium/genética , Cryptosporidium/clasificación , Humanos , ADN Protozoario/genética , Secuenciación de Nucleótidos de Alto Rendimiento , Código de Barras del ADN Taxonómico/métodos , Criptosporidiosis/parasitología , Variación GenéticaRESUMEN
Cryptosporidium infection is a common occurrence in rodents worldwide. In this study, 435 wild brown rats were captured from an animal feedlot in Xinjiang, China, with a fecal sample obtained directly from the rectal contents of each rat. The DNA extracted from these fecal samples was analyzed for Cryptosporidium spp. using PCR targeting the SSU rRNA gene. The prevalence of Cryptosporidium infection in brown rats was found to be 5.5% (24 out of 435). Interestingly, the infection rates varied among different animal enclosures, with rates of 0% in the chicken coop (0/51), cowshed (0/3), and varying rates in other areas including the sheepfold (6.1%, 6/98), the pigsty (7.6%, 10/132), the dovecote (7.0%, 5/71), and outdoor environments (3.8%, 3/80). The study identified three species and one genotype of Cryptosporidium, namely C. occultus (n = 10), C. parvum (n = 4), C. ditrichi (n = 1), and Cryptosporidium rat genotype IV (n = 9). Additionally, two of the C. parvum isolates were successfully subtyped as IIdA19G1 (n = 2) at the gp60 gene. These results offer valuable insights into the prevalence and genetic diversity of Cryptosporidium in brown rats within the region.
Asunto(s)
Criptosporidiosis , Cryptosporidium , Heces , Animales , Cryptosporidium/genética , Cryptosporidium/clasificación , Cryptosporidium/aislamiento & purificación , Criptosporidiosis/parasitología , Criptosporidiosis/epidemiología , China/epidemiología , Ratas/parasitología , Heces/parasitología , Prevalencia , Genotipo , ADN Protozoario/genética , Filogenia , Enfermedades de los Roedores/parasitología , Enfermedades de los Roedores/epidemiología , Reacción en Cadena de la PolimerasaRESUMEN
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.
Asunto(s)
Criptosporidiosis , Cryptosporidium , Erizos , Filogenia , Animales , Criptosporidiosis/parasitología , Criptosporidiosis/epidemiología , Cryptosporidium/genética , Cryptosporidium/clasificación , Cryptosporidium/aislamiento & purificación , ADN Protozoario/genética , ADN Ribosómico/genética , ADN Ribosómico/química , Heces/parasitología , Genotipo , Erizos/parasitología , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , ARN Ribosómico 18S/genética , Análisis de Secuencia de ADN , EspañaRESUMEN
Non-human primates (NHPs) are the group that most share infectious agents with humans due to their close taxonomic relationship. The southern brown howler monkeys (Alouatta guariba clamitans) are endemic primates from Brazil and Argentina's Atlantic Forest. This study aimed to investigate the presence of intestinal parasites in free-living (FL) and captive (CA) southern brown howler monkeys. Thirty-nine stool samples were collected in two areas in southern Brazil, 15 FL and 24 CA. Stool sediments obtained by centrifugal sedimentation technique were used for microscopic analysis and direct immunofluorescence assay and evaluated by molecular analysis through amplification and sequencing of TPI fragments. Intestinal parasites Giardia duodenalis, Cryptosporidium spp., and Trypanoxyuris minutus were detected at coproparasitological analysis. This is the first report of the presence of Cryptosporidium spp. in free-living howlers. The molecular characterization of G. duodenalis isolates indicated assemblage B for the first time found in free-living A. guariba clamitans. The high prevalence of G. duodenalis transmission in CA howler monkeys can be explained by direct contact with humans and frequent soil contact. The presence of a potentially zoonotic assemblage in these animals indicates that the process of fragmentation and cohabitation with humans and livestock affects the wildlife, thus indicating a need for eco-health measures.
Asunto(s)
Alouatta , Giardia lamblia , Giardiasis , Enfermedades de los Monos , Animales , Alouatta/parasitología , Brasil/epidemiología , Enfermedades de los Monos/parasitología , Enfermedades de los Monos/epidemiología , Giardiasis/veterinaria , Giardiasis/parasitología , Giardiasis/epidemiología , Giardia lamblia/aislamiento & purificación , Giardia lamblia/genética , Giardia lamblia/clasificación , Heces/parasitología , Animales de Zoológico/parasitología , Cryptosporidium/aislamiento & purificación , Cryptosporidium/clasificación , Cryptosporidium/genética , Prevalencia , Masculino , Animales Salvajes/parasitología , Femenino , Criptosporidiosis/parasitología , Criptosporidiosis/epidemiologíaRESUMEN
Cryptosporidium spp. are protozoa commonly found in domestic and wild animals. Limited information is available on Cryptosporidium in deer worldwide. In this study, 201 fecal samples were collected from Alpine musk deer on three farms in Gansu Province, China. Detection and subtyping of Cryptosporidium were performed by PCR and sequence analysis of the SSU rRNA and gp60 genes. The prevalence of Cryptosporidium infection in Alpine musk deer was 3.9% (8/201), with infection rates of 1.0% (1/100), 2.8% (1/36), and 9.2% (6/65) in three different farms. All positive samples for Cryptosporidium were from adult deer. Two Cryptosporidium species were identified, including C. parvum (n = 2) and C. xiaoi (n = 6). The C. parvum isolates were subtyped as IIdA15G1, while the C. xiaoi isolates were subtyped as XXIIIa (n = 2) and XXIIIg (n = 4). The IIdA15G1 subtype of C. parvum was found for the first time in deer. These results provide important insights into the identity and human infectious potential of Cryptosporidium in farmed Alpine musk deer.
Asunto(s)
Criptosporidiosis , Cryptosporidium , Ciervos , Heces , Animales , Ciervos/parasitología , Criptosporidiosis/parasitología , Criptosporidiosis/epidemiología , Cryptosporidium/genética , Cryptosporidium/aislamiento & purificación , Cryptosporidium/clasificación , China/epidemiología , Heces/parasitología , Prevalencia , ADN Protozoario/genética , Filogenia , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADN , Genotipo , ADN Ribosómico/genética , ADN Ribosómico/químicaRESUMEN
Cryptosporidiosis is an infectious enteric disease caused by species (some of them zoonotic) of the genus Cryptosporidium that in many countries are under surveillance. Typing assays critical to the surveillance of cryptosporidiosis typically involve characterization of Cryptosporidium glycoprotein 60 genes (gp60). Here, we characterized the gp60 of Cryptosporidium suis from two samples-a human and a porcine faecal sample-based on which a preliminary typing scheme was developed. A conspicuous feature of the C. suis gp60 was a novel type of tandem repeats located in the 5' end of the gene and that took up 777/1635 bp (48%) of the gene. The C. suis gp60 lacked the classical poly-serine repeats (TCA/TCG/TCT), which is usually subject to major genetic variation, and the length of the tandem repeat made a typing assay incorporating this region based on Sanger sequencing practically unfeasible. We therefore designed a typing assay based on the post-repeat region only and applied it to C. suis-positive samples from suid hosts from Norway, Denmark, and Spain. We were able to distinguish three different subtypes; XXVa-1, XXVa-2, and XXVa-3. Subtype XXVa-1 had a wider geographic distribution than the other subtypes and was also observed in the human sample. We think that the present data will inform future strategies to develop a C. suis typing assay that could be even more informative by including a greater part of the gene, including the tandem repeat region, e.g., by the use of long-read next-generation sequencing.
Asunto(s)
Criptosporidiosis , Cryptosporidium , Secuencias Repetidas en Tándem , Animales , Criptosporidiosis/parasitología , Criptosporidiosis/epidemiología , Porcinos , Humanos , Cryptosporidium/genética , Cryptosporidium/clasificación , Filogenia , Enfermedades de los Porcinos/parasitología , Proteínas Protozoarias/genética , Heces/parasitologíaRESUMEN
Cryptosporidium species, causing diarrheal illnesses in humans and animals worldwide, are under investigation for their molecular epidemiology in Japan. The study focuses on detecting Cryptosporidium species in humans, animals, water, and the environment, revealing three species in people: C. parvum, C. meleagridis, and C. hominis. Subtype IIa of the C. parvum gp60 gene is prevalent, indicating potential zoonotic transmission. Animal studies identified sixteen species, mainly cattle and pets, with C. parvum (subtype IIa) common in cattle and C. canis and C. felis prevalent in pets. Additionally, C. bovis and C. ryanae were found in cattle and sika deer. Knowledge gaps exist, particularly in water and environmental source typing, with limited research revealing five species and five genotypes, suggesting a significant role of water in transmission. Further research is needed to understand the molecular diversity and transmission dynamics across humans, animals, water, and the environment in Japan.
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Criptosporidiosis , Cryptosporidium , Epidemiología Molecular , Animales , Japón/epidemiología , Criptosporidiosis/epidemiología , Criptosporidiosis/parasitología , Criptosporidiosis/transmisión , Cryptosporidium/genética , Cryptosporidium/clasificación , Cryptosporidium/aislamiento & purificación , Humanos , Bovinos , Zoonosis/epidemiología , Zoonosis/parasitología , Zoonosis/transmisión , Genotipo , Enfermedades de los Bovinos/epidemiología , Enfermedades de los Bovinos/parasitología , Enfermedades de los Bovinos/transmisión , Ciervos/parasitología , Mascotas/parasitologíaRESUMEN
BACKGROUND: Enteric parasitic infections remain a major public health problem globally. Cryptosporidium spp., Cyclospora spp. and Giardia spp. are parasites that cause diarrhea in the general populations of both developed and developing countries. Information from molecular genetic studies on the speciation of these parasites and on the role of animals as vectors in disease transmission is lacking in Ghana. This study therefore investigated these diarrhea-causing parasites in humans, domestic rats and wildlife animals in Ghana using molecular tools. METHODS: Fecal samples were collected from asymptomatic school children aged 9-12 years living around the Shai Hills Resource Reserve (tourist site), from wildlife (zebras, kobs, baboons, ostriches, bush rats and bush bucks) at the same site, from warthogs at the Mole National Park (tourist site) and from rats at the Madina Market (a popular vegetable market in Accra, Ghana. The 18S rRNA gene (18S rRNA) and 60-kDa glycoprotein gene (gp60) for Cryptosporidium spp., the glutamate dehydrogenase gene (gdh) for Giardia spp. and the 18S rDNA for Cyclospora spp. were analyzed in all samples by PCR and Sanger sequencing as markers of speciation and genetic diversity. RESULTS: The parasite species identified in the fecal samples collected from humans and animals included the Cryptosporidium species C. hominis, C. muris, C. parvum, C. tyzzeri, C. meleagridis and C. andersoni; the Cyclopora species C. cayetanensis; and the Gardia species, G. lamblia and G. muris. For Cryptosporidium, the presence of the gp60 gene confirmed the finding of C. parvum (41%, 35/85 samples) and C. hominis (29%, 27/85 samples) in animal samples. Cyclospora cayetanensis was found in animal samples for the first time in Ghana. Only one human sample (5%, 1/20) but the majority of animal samples (58%, 51/88) had all three parasite species in the samples tested. CONCLUSIONS: Based on these results of fecal sample testing for parasites, we conclude that animals and human share species of the three genera (Cryptosporidium, Cyclospora, Giardia), with the parasitic species mostly found in animals also found in human samples, and vice-versa. The presence of enteric parasites as mixed infections in asymptomatic humans and animal species indicates that they are reservoirs of infections. This is the first study to report the presence of C. cayetanensis and C. hominis in animals from Ghana. Our findings highlight the need for a detailed description of these parasites using high-throughput genetic tools to further understand these parasites and the neglected tropical diseases they cause in Ghana where such information is scanty.
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Animales Domésticos , Animales Salvajes , Criptosporidiosis , Cryptosporidium , Cyclospora , Ciclosporiasis , Heces , Animales , Ghana/epidemiología , Cyclospora/genética , Cyclospora/aislamiento & purificación , Cyclospora/clasificación , Cryptosporidium/genética , Cryptosporidium/aislamiento & purificación , Cryptosporidium/clasificación , Heces/parasitología , Ciclosporiasis/epidemiología , Ciclosporiasis/parasitología , Ciclosporiasis/veterinaria , Animales Salvajes/parasitología , Criptosporidiosis/parasitología , Criptosporidiosis/epidemiología , Criptosporidiosis/transmisión , Humanos , Niño , Animales Domésticos/parasitología , Ratas , ADN Protozoario/genética , ARN Ribosómico 18S/genética , Giardiasis/veterinaria , Giardiasis/parasitología , Giardiasis/epidemiología , Diarrea/parasitología , Diarrea/veterinaria , Diarrea/epidemiología , Filogenia , Giardia/genética , Giardia/aislamiento & purificación , Giardia/clasificaciónRESUMEN
Cryptosporidium spp. are significant zoonotic intestinal parasites that induce diarrhea and even death across most vertebrates, including humans. Previous studies showed that sheep are important hosts for Cryptosporidium and that its distribution in sheep is influenced by geography, feeding patterns, age, and season. Environmental factors also influence the transmission of Cryptosporidium. Molecular studies of Cryptosporidium in sheep have been conducted in only a few regions of China, and studies into the effect of sheep-housing environments on Cryptosporidium transmission are even rarer. To detect the prevalence of Cryptosporidium in large-scale sheep-housing farms, a total of 1241 fecal samples were collected from sheep, 727 environmental samples were taken from sheep housing, and 30 water samples were collected in six regions of China. To ascertain the existence of the parasite and identify the species of Cryptosporidium spp., we conducted nested PCR amplification of DNA extracted from all samples using the small-subunit (SSU) rRNA gene as a target. For a more in-depth analysis of Cryptosporidium spp. subtypes, C. xiaoi-and C. ubiquitum-positive samples underwent separate nested PCR amplification targeting the 60 kDa glycoprotein (gp60) gene. The amplification of the Cryptosporidium spp. SSU rRNA gene locus from the whole genomic DNA of all samples yielded a positive rate of 1.2% (20/1241) in fecal samples, 0.1% (1/727) in environmental samples, and no positive samples were found in water samples. The prevalence of Cryptosporidium spp. infection in large-scale housed sheep was 1.7%, which was higher than that in free-ranging sheep (0.0%). The highest prevalence of infection was found in weaning lambs (6.8%). Among the different seasons, the peaks were found in the fall and winter. The most prevalent species were C. xiaoi and C. ubiquitum, with the former accounting for the majority of infections. The distribution of C. xiaoi subtypes was diverse, with XXIIIc (n = 1), XXIIId (n = 2), XXIIIe (n = 2), and XXIIIl (n = 4) identified. In contrast, only one subtype, XIIa (n = 9), was found in C. ubiquitum. In this study, C. xiaoi and C. ubiquitum were found to be the predominant species, and Cryptosporidium was found to be present in the environment. These findings provide an important foundation for the comprehensive prevention and management of Cryptosporidium in intensively reared sheep. Furthermore, by elucidating the prevalence of Cryptosporidium in sheep and its potential role in environmental transmission, this study deepens our understanding of the intricate interactions between animal health, environmental contamination, and public health dynamics.
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Criptosporidiosis , Cryptosporidium , Granjas , Heces , Variación Genética , Enfermedades de las Ovejas , Animales , Cryptosporidium/genética , Cryptosporidium/aislamiento & purificación , Cryptosporidium/clasificación , Ovinos/parasitología , China/epidemiología , Criptosporidiosis/epidemiología , Criptosporidiosis/parasitología , Criptosporidiosis/transmisión , Enfermedades de las Ovejas/epidemiología , Enfermedades de las Ovejas/parasitología , Enfermedades de las Ovejas/transmisión , Prevalencia , Heces/parasitología , FilogeniaRESUMEN
As data accumulate in GenBank, the difficulties of delineating species of Cryptosporidium based on nuclear small subunit ribosomal RNA (ssu rRNA) gene information alone becomes increasingly evident. Here, we summarize currently available evidence suggesting that several ssu rDNA sequences primarily referred to as Cryptosporidium suis (some of them from non-suid hosts) should be considered Cryptosporidium occultus.
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Criptosporidiosis , Cryptosporidium , ADN Ribosómico , Cryptosporidium/genética , Cryptosporidium/clasificación , Cryptosporidium/aislamiento & purificación , Criptosporidiosis/parasitología , Animales , ADN Ribosómico/genética , ADN Protozoario/genética , Humanos , Filogenia , Análisis de Secuencia de ADN/métodosRESUMEN
Cryptosporidium spp. is an important foodborne and waterborne pathogen in humans and animals, causing diarrhoea in humans and respiratory and gastrointestinal diseases in birds. However, reports of Cryptosporidium infection in bar-headed goose are limited. To determine the infection rate and species/genotypes of Cryptosporidium in bar-headed goose in China, a total of 358 fecal samples were collected from 3 regions. Nested PCR was used to amplify Cryptosporidium SSU rRNA regions from the fecal extracted-DNA samples. The total infection rate of Cryptosporidium in bar-headed in China was 3.9 % (14/358), with 4.2 % (5/120) in Aba (Ngawa) Tibetan and Qiang Autonomous Prefect, Sichuan province, 7.6 % (9/119) in Maqu county, Gansu province, and 0.0 % (0/119) in Caohai, Wei ning county, Guizhou province. The differences in prevalence rate by region were statistically significant. All positive samples were identified as Cryptosporidium goose genotype I (n = 14). This is the first systematic investigation of the epidemiological status and dominant species/genotypes of Cryptosporidium in bar-headed goose in China, thereby enhancing our understanding of the epidemiology of Cryptosporidium infection in wild migratory birds.
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Criptosporidiosis , Cryptosporidium , Heces , Gansos , Genotipo , Animales , Cryptosporidium/genética , Cryptosporidium/aislamiento & purificación , Cryptosporidium/clasificación , Criptosporidiosis/epidemiología , Criptosporidiosis/parasitología , China/epidemiología , Prevalencia , Gansos/parasitología , Heces/parasitología , Reacción en Cadena de la Polimerasa/veterinaria , Enfermedades de las Aves de Corral/epidemiología , Enfermedades de las Aves de Corral/parasitología , ADN Protozoario/análisisRESUMEN
Cryptosporidium spp., Enterocytozoon bieneusi, and Giardia duodenalis are common intestinal pathogens that infect humans and animals. To date, research regarding these three protozoa in the Ningxia Hui Autonomous Region (Ningxia) has mostly been limited to a single pathogen, and comprehensive data on mixed infections are unavailable. This study aimed to evaluate the zoonotic potential of these three protozoa. In this study, small subunit ribosomal RNA (SSU rRNA) and 60 kDa glycoprotein (gp60) genes of Cryptosporidium; internal transcribed spacer (ITS) gene of E. bieneusi; and SSU rRNA, glutamate dehydrogenase (gdh), triosephosphate isomerase (tpi), and beta-giardin (bg) genes of G. duodenalis were examined. DNA extraction, polymerase chain reaction, and sequence analysis were performed on fecal samples collected from 320 dairy cattle at three intensive dairy farms in Ningxia in 2021 to determine the prevalence and genetic characteristics of these three protozoa. The findings revealed that 61.56% (197/320) of the samples were infected with at least one protozoan. The overall prevalence of Cryptosporidium was 19.38% (62/320), E. bieneusi was 41.56% (133/320), and G. duodenalis was 29.38% (94/320). This study identified four Cryptosporidium species (C. bovis, C. andersoni, C. ryanae, and C. parvum) and the presence of mixed infections with two or three Cryptosporidium species. C. bovis was the dominant species in this study, while the dominant C. parvum subtypes were IIdA15G1 and IIdA20G1. The genotypes of E. bieneusis were J, BEB4, and I alongside the novel genotypes NX1-NX8, all belonging to group 2, with genotype J being dominant. G. duodenalis assemblages were identified as assemblages E, A, and B, and a mixed infection involving assemblages A + E was identified, with assemblage E being the dominant one. Concurrently, 11 isolates formed 10 different assemblage E multilocus genotypes (MLGs) and 1 assemblage A MLG and assemblage E MLGs formed 5 subgroups. To the best of our knowledge, this is the first report on mixed infection with two or three Cryptosporidium species in cattle in Ningxia and on the presence of the C. parvum subtype IIdA20G1 in this part of China. This study also discovered nine genotypes of E. bieneusis and novel features of G. duodenalis assemblages in Ningxia. This study indicates that dairy cattle in this region may play a significant role in the zoonotic transmission of Cryptosporidium spp., E. bieneusi, and G. duodenalis.
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Enfermedades de los Bovinos , Criptosporidiosis , Cryptosporidium , Enterocytozoon , Giardia lamblia , Giardiasis , Microsporidiosis , Animales , Enterocytozoon/genética , Enterocytozoon/aislamiento & purificación , Bovinos , Cryptosporidium/genética , Cryptosporidium/aislamiento & purificación , Cryptosporidium/clasificación , Giardia lamblia/genética , Giardia lamblia/aislamiento & purificación , Criptosporidiosis/epidemiología , Criptosporidiosis/parasitología , China/epidemiología , Prevalencia , Microsporidiosis/veterinaria , Microsporidiosis/epidemiología , Enfermedades de los Bovinos/epidemiología , Enfermedades de los Bovinos/parasitología , Enfermedades de los Bovinos/microbiología , Giardiasis/veterinaria , Giardiasis/epidemiología , Giardiasis/parasitología , Femenino , Heces/parasitología , Heces/microbiologíaRESUMEN
Bats stand as one of the most diverse groups in the animal kingdom and are key players in the global transmission of emerging pathogens. However, their role in transmitting Enterocytozoon bieneusi and Cryptosporidium spp. remains unclear. This study aimed to evaluate the occurrence and genetic diversity of the two pathogens in fruit bats (Rousettus leschenaultii) in Hainan, China. Ten fresh fecal specimens of fruit bats were collected from Wanlvyuan Gardens, Haikou, China. The fecal samples were tested for E. bieneusi and Cryptosporidium spp. using Polymerase Chain Reaction (PCR) analysis and sequencing the internal transcribed spacer (ITS) region and partial small subunit of ribosomal RNA (SSU rRNA) gene, respectively. Genetic heterogeneity across Cryptosporidium spp. isolates was assessed by sequencing 4 microsatellite/minisatellite loci (MS1, MS2, MS3, and MS16). The findings showed that out of the ten specimens analyzed, 2 (20 %) and seven (70.0 %) were tested positive for E. bieneusi and Cryptosporidium spp., respectively. DNA sequence analysis revealed the presence of two novel Cryptosporidium genotypes with 94.4 to 98.6 % sequence similarity to C. andersoni, named as Cryptosporidium bat-genotype-XXI and bat-genotype-XXII. Three novel sequences of MS1, MS2 and MS16 loci identified here had 95.4 to 96.9 % similarity to the known sequences, which were deposited in the GenBank. Two genotypes of E. bieneusi were identified, including a novel genotype named HNB-I and a zoonotic genotype PigEbITS7. The discovery of these novel sequences provides meaningful data for epidemiological studies of the both pathogens. Meanwhile our results are also presented that the fruit bats infected with E. bieneusi, but not with Cryptosporidium, should be considered potential public health threats.
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Quirópteros , Criptosporidiosis , Cryptosporidium , Enterocytozoon , Heces , Genotipo , Microsporidiosis , Animales , Quirópteros/parasitología , Quirópteros/microbiología , Enterocytozoon/genética , Enterocytozoon/aislamiento & purificación , Enterocytozoon/clasificación , Cryptosporidium/genética , Cryptosporidium/clasificación , Cryptosporidium/aislamiento & purificación , China/epidemiología , Microsporidiosis/veterinaria , Microsporidiosis/epidemiología , Microsporidiosis/parasitología , Microsporidiosis/microbiología , Criptosporidiosis/parasitología , Criptosporidiosis/epidemiología , Heces/parasitología , Heces/microbiología , Variación Genética , Filogenia , Análisis de Secuencia de ADN , ADN Espaciador Ribosómico/genética , Reacción en Cadena de la Polimerasa , ADN de Hongos/genética , Repeticiones de Microsatélite , ADN Protozoario/genética , Parques RecreativosRESUMEN
Cryptosporidia (Cryptosporidium) is a protozoan that is widely parasitic in the intestinal cells of humans and animals, and it is also an important zoonotic parasite. However, there is no epidemiological investigation on Cryptosporidium spp. infection in infants with diarrhea of Inner Mongolia, the largest livestock region in China. To investigate the prevalence of Cryptosporidium, 2435 fresh fecal samples were collected from children with diarrhea in Inner Mongolia Maternal and Child Health Care Hospital. Molecular characterization of Cryptosporidium was carried out based on its 18S rRNA and gp60 gene sequences. The overall prevalence was 12.85% (313/2435), and in Hohhot (12.15%), it was lower than that in the surrounding city (14.87%) (P < 0.05). Moreover, Cryptosporidium was detected in different seasons and sexes. Concerning the age of children with diarrhea, the prevalence of those age groups between 0 and 1 was obviously lower than others, and there were significant differences in the prevalence at different ages (P < 0.001). Analysis of the 18S rRNA gene sequence revealed that all the positive samples were Cryptosporidium parvum, and there were 5 subtypes (IIdA23G3, IIdA24G3, IIdA24G4, IIdA25G3, and IIdA25G4). To the best of our knowledge, the above subtypes have not been reported. Our results provide a relevant basis for control and education on food safety and foodborne illness prevention.