RESUMO
Diarrhea caused by zoonotic pathogens is one of the most common diseases in dairy calves, threatening the health of young animals. Humans are also at risk, in particular children. To explore the pathogens causing diarrhea in dairy calves, the present study applied PCR-based sequencing tools to investigate the occurrence and molecular characteristics of three parasites (Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi) and three bacterial pathogens (Escherichia coli, Clostridium perfringens, and Salmonella spp.) in 343 fecal samples of diarrheic dairy calves from five farms in Lingwu County, Ningxia Hui Autonomous Region, China. The total positive rate of these pathogens in diarrheic dairy calves was 91.0% (312/343; 95% CI, 87.9-94.0), with C. perfringens (61.5%, 211/343; 95% CI, 56.3-66.7) being the dominant one. Co-infection with two to five pathogens was found in 67.3% (231/343; 95% CI, 62.4-72.3) of investigated samples. There were significant differences (p < 0.05) in the positive rates of Cryptosporidium spp. and diarrheagenic E. coli among farms, age groups, and seasons. Two Cryptosporidium species (C. parvum and C. bovis) and five gp60 subtypes of C. parvum (IIdA15G1, IIdA20G1, IIdA19G1, IIdA14G1, and a novel IIdA13G1) were identified. Two assemblages (assemblage E and zoonotic assemblage A) of G. duodenalis and six ITS genotypes of E. bieneusi (J, Henan-IV, EbpC, I, EbpA, and ESH-01) were observed. Four virulence genes (eaeA, stx1, stx2, and st) of diarrheagenic E. coli and one toxin type (type A) of C. perfringens were detected. Our study enriches our knowledge on the characteristics and zoonotic potential of diarrhea-related pathogens in dairy calves.
Title: Caractérisation moléculaire des protozoaires parasites zoonotiques courants et des bactéries responsables de diarrhée chez les veaux laitiers dans la région autonome Hui du Ningxia, en Chine. Abstract: La diarrhée causée par des agents pathogènes zoonotiques est l'une des maladies les plus courantes chez les veaux laitiers, menaçant la santé des jeunes animaux. Ceci est également un risque pour la santé humaine, en particulier les enfants. Pour explorer les agents pathogènes responsables de la diarrhée chez les veaux laitiers, cette étude a utilisé des outils de séquençage basés sur la PCR pour étudier l'occurrence et les caractères moléculaires de trois parasites (Cryptosporidium spp., Giardia duodenalis et Enterocytozoon bieneusi) et de trois agents pathogènes bactériens (Escherichia coli, Clostridium perfringens et Salmonella spp.) dans 343 échantillons fécaux de veaux laitiers diarrhéiques provenant de cinq fermes du comté de Lingwu, région autonome Hui du Ningxia, en Chine. Le taux total positif de ces pathogènes chez les veaux laitiers diarrhéiques était de 91,0 % (312/343; IC à 95 %, 87,994,0), et C. perfringens (61,5 %, 211/343; IC à 95 %, 56,366,7) était le plus répandu. Une co-infection avec deux à cinq pathogènes a été trouvée dans 67,3 % (231/343; IC à 95 %, 62,472,3) des échantillons étudiés. Il y avait des différences significatives (p < 0,05) dans les taux positifs de Cryptosporidium spp. et d'E. coli diarrhéogènes entre les fermes, les groupes d'âge et les saisons. Deux espèces de Cryptosporidium (C. parvum et C. bovis) et cinq sous-types de gp60 de C. parvum (IIdA15G1, IIdA20G1, IIdA19G1, IIdA14G1 et un nouveau, IIdA13G1) ont été identifiés. Deux assemblages (assemblage E et assemblage zoonotique A) de G. duodenalis et six génotypes ITS d'E. bieneusi (J, Henan-IV, EbpC, I, EbpA et ESH-01) ont été observés. Quatre gènes de virulence (eaeA, stx1, stx2 et st) d'E. coli diarrhéogènes et un type de toxine (type A) de C. perfringens ont été détectés. Notre étude enrichit les connaissances sur les caractères et le potentiel zoonotique des agents pathogènes liés à la diarrhée chez les veaux laitiers.
Assuntos
Doenças dos Bovinos , Criptosporidiose , Cryptosporidium , Diarreia , Enterocytozoon , Fezes , Giardia lamblia , Zoonoses , Animais , Bovinos , Diarreia/veterinária , Diarreia/parasitologia , Diarreia/microbiologia , Diarreia/epidemiologia , China/epidemiologia , Doenças dos Bovinos/parasitologia , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/microbiologia , Cryptosporidium/genética , Cryptosporidium/isolamento & purificação , Cryptosporidium/classificação , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , Enterocytozoon/classificação , Giardia lamblia/genética , Giardia lamblia/isolamento & purificação , Giardia lamblia/classificação , Fezes/parasitologia , Fezes/microbiologia , Criptosporidiose/epidemiologia , Criptosporidiose/parasitologia , Escherichia coli/isolamento & purificação , Escherichia coli/genética , Escherichia coli/classificação , Giardíase/veterinária , Giardíase/epidemiologia , Giardíase/parasitologia , Coinfecção/veterinária , Coinfecção/epidemiologia , Coinfecção/parasitologia , Coinfecção/microbiologia , Microsporidiose/veterinária , Microsporidiose/epidemiologia , Clostridium perfringens/isolamento & purificação , Clostridium perfringens/genética , Clostridium perfringens/classificação , Salmonella/isolamento & purificação , Salmonella/genética , Salmonella/classificação , Humanos , Reação em Cadeia da Polimerase/veterinária , Indústria de LaticíniosRESUMO
Enterocytozoon bieneusi is a zoonotic pathogen prevalent in mammalian and avian hosts across the globe. Wild small mammals, being abundant worldwide, serve as important sources of zoonotic disease transmission to humans. Here, 227 fecal samples were collected from five rodent and shrew species on 34 pig farms in China to investigate the prevalence and molecular characterization of E. bieneusi. The overall prevalence of E. bieneusi was 17.18% (39/227), with a distribution of 23.53% (32/136) in Rattus tanezumi, 8.62% (5/58) in Rattus norvegicus, and 8.00% (2/25) in Mus musculus. Eight E. bieneusi genotypes were identified, comprising four known genotypes: D (n = 8), EbpC (n = 8), PigEBITS7 (n = 9), and EbpA (n = 2), and four novel genotypes: CHPR1 (n = 7), CHPR2 (n = 1), CHPR3 (n = 2), and CHPR4 (n = 2). This study is the first to report E. bieneusi in rodents from pig farms in Henan, Shaanxi, and Shanxi Provinces in China. The host range of genotype EbpC was expanded with its first detection in M. musculus and R. tanezumi. All identified E. bieneusi genotypes belong to group 1, raising concerns about these sympatric rodents being reservoirs of zoonotic transmission. Moreover, the widespread distribution of genotype EbpC suggests potential cross-species transmission between sympatric rodents and domestic pigs. Our findings highlight the potential role of sympatric rodents in facilitating the spillover of E. bieneusi from pig farms, which could pose a potential public health threat.
Title: Les rongeurs sympatriques sauvages vivant dans les élevages porcins peuvent faciliter la propagation d'Enterocytozoon bieneusi. Abstract: Enterocytozoon bieneusi est un pathogène zoonotique répandu mondialement chez les hôtes mammifères et aviaires. Les petits mammifères sauvages, abondants dans le monde entier, constituent d'importantes sources de transmission de maladies zoonotiques à l'homme. Ici, 227 échantillons fécaux ont été collectés auprès de cinq espèces de rongeurs et de musaraignes dans 34 élevages porcins en Chine pour étudier la prévalence et la caractérisation moléculaire d'E. bieneusi. La prévalence globale d'E. bieneusi était de 17,18 % (39/227), avec une distribution de 23,53 % (32/136) chez Rattus tanezumi, 8,62 % (5/58) chez Rattus norvegicus et 8,00 % (2/25) chez Mus musculus. Huit génotypes d'E. bieneusi ont été identifiés, dont quatre génotypes connus (D (n = 8), EbpC (n = 8), PigEBITS7 (n = 9) et EbpA (n = 2)) et quatre génotypes nouveaux (CHPR1 (n = 7), CHPR2 (n = 1), CHPR3 (n = 2) et CHPR4 (n = 2)). Cette étude est la première à signaler la présence d'E. bieneusi chez des rongeurs provenant d'élevages porcins des provinces du Henan, du Shaanxi et du Shanxi en Chine. La gamme d'hôtes du génotype EbpC a été élargie avec sa première détection chez M. musculus et R. tanezumi. Tous les génotypes d'E. bieneusi identifiés appartiennent au groupe 1, ce qui soulève des inquiétudes quant au fait que ces rongeurs sympatriques soient des réservoirs de transmission zoonotique. De plus, la large distribution du génotype EbpC suggère une transmission interspécifique potentielle entre les rongeurs sympatriques et les porcs domestiques. Nos résultats soulignent le rôle potentiel des rongeurs sympatriques dans la facilitation de la propagation d'E. bieneusi à partir des élevages porcins, ce qui pourrait constituer une menace potentielle pour la santé publique.
Assuntos
Enterocytozoon , Fazendas , Fezes , Genótipo , Microsporidiose , Roedores , Doenças dos Suínos , Zoonoses , Animais , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , Enterocytozoon/classificação , Microsporidiose/veterinária , Microsporidiose/epidemiologia , Microsporidiose/transmissão , Microsporidiose/microbiologia , China/epidemiologia , Suínos , Fezes/microbiologia , Roedores/microbiologia , Ratos , Camundongos , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/microbiologia , Doenças dos Suínos/transmissão , Zoonoses/transmissão , Zoonoses/epidemiologia , Prevalência , Filogenia , Doenças dos Roedores/epidemiologia , Doenças dos Roedores/microbiologia , Doenças dos Roedores/transmissão , Musaranhos/microbiologia , Musaranhos/parasitologia , Humanos , Animais Selvagens/microbiologia , Reservatórios de Doenças/microbiologia , Reservatórios de Doenças/veterináriaRESUMO
Although brought to the forefront in the 1980s with the AIDS pandemic, microsporidia infecting humans are still little known. Enterocytozoon bieneusi, by far the most frequent microsporidia species causing diseases in humans, is responsible for intestinal illness in both non- and immunocompromised patients. This species presents an astonishing genetic diversity with more than 500 genotypes described, some of which have a strong zoonotic potential. Indeed, E. bieneusi infects a broad array of hosts, from wild to domestic animals. This emerging eukaryotic pathogen has thus been associated with foodborne/waterborne outbreaks. Several molecular assays have been developed to enhance its diagnosis or for epidemiological purposes, providing valuable new data. Here, we propose an overview of the current knowledge on this major species among the microsporidia, so far rather neglected in human medicine.
Assuntos
Enterocytozoon , Microsporidiose , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , Enterocytozoon/classificação , Microsporidiose/veterinária , Microsporidiose/epidemiologia , Microsporidiose/microbiologia , Humanos , Animais , Zoonoses/microbiologia , Genótipo , Variação GenéticaRESUMO
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.
Assuntos
Chinchila , Cryptosporidium , Encephalitozoon , Encefalitozoonose , Enterocytozoon , Giardia lamblia , Giardíase , Microsporidiose , Animais de Estimação , Zoonoses , Animais , Chinchila/parasitologia , Encephalitozoon/genética , Encephalitozoon/isolamento & purificação , Encephalitozoon/classificação , Zoonoses/parasitologia , Cryptosporidium/genética , Cryptosporidium/classificação , Cryptosporidium/isolamento & purificação , Giardíase/veterinária , Giardíase/parasitologia , Giardia lamblia/genética , Giardia lamblia/isolamento & purificação , Giardia lamblia/classificação , República Tcheca/epidemiologia , Encefalitozoonose/veterinária , Encefalitozoonose/epidemiologia , Encefalitozoonose/microbiologia , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , Microsporidiose/veterinária , Microsporidiose/epidemiologia , Criptosporidiose/parasitologia , Criptosporidiose/epidemiologia , RNA Ribossômico 18S/genética , Fezes/parasitologia , Fezes/microbiologia , MasculinoRESUMO
Nowadays, the use of qPCR for the diagnosis of intestinal microsporidiosis is increasing. There are several studies on the evaluation of qPCR performance but very few focus on the stool pretreatment step before DNA extraction, which is nevertheless a crucial step. This study focuses on the mechanical pretreatment of stools for Enterocytozoon bieneusi spores DNA extraction. Firstly, a multicenter comparative study was conducted evaluating seven extraction methods (manual or automated) including various mechanical pretreatment. Secondly, several durations and grinding speeds and types of beads were tested in order to optimize mechanical pretreatment. Extraction methods of the various centers had widely-varying performances especially for samples with low microsporidia loads. Nuclisens® easyMAG (BioMérieux) and Quick DNA Fecal/Soil Microbe Microprep kit (ZymoResearch) presented the best performances (highest frequencies of detection of low spore concentrations and lowest Ct values). Optimal performances of mechanical pretreatment were obtained by applying a speed of 30 Hz during 60 s with the TissueLyser II (Qiagen) using commercial beads of various materials and sizes (from ZymoResearch or MP Biomedicals). Overall, the optimal DNA extraction method for E. bieneusi spores contained in stool samples was obtained with a strong but short bead beating using small-sized beads from various materials.
Assuntos
DNA Fúngico , Enterocytozoon , Fezes , Microsporidiose , Fezes/microbiologia , Enterocytozoon/isolamento & purificação , Enterocytozoon/genética , Humanos , Microsporidiose/diagnóstico , Microsporidiose/microbiologia , DNA Fúngico/isolamento & purificação , DNA Fúngico/genética , DNA Fúngico/análise , Manejo de Espécimes/métodos , Esporos Fúngicos/isolamento & purificação , Esporos Fúngicos/genética , Reação em Cadeia da Polimerase em Tempo Real/métodosRESUMO
Objective: In patients with end-stage kidney disease, kidney transplantation is the kidney replacement therapy option that provides the most successful survival. However, immunosuppression agents administered after kidney transplantation can increase the risk of opportunistic infections. Microsporidia are obligate intracellular pathogens that can be fatal in immunosuppressed patients. The present study aimed to determine the prevalence of microsporidia in kidney transplantation recipients and the molecular characterization of the detected species. Methods: To evaluate the prevalence of renal microsporidiosis in kidney transplant recipients, the urine samples from a total of 325 patients were analyzed by real-time and nested polymerase chain reaction for Encephalitozoon spp. and Enterocytozoon bieneusi. Results: Only one (0.4%) sample from the adult patient was positive for the Encephalitozoon species, while no positivity was found in pediatric patients. It was determined as Encephalitozoon intestinalis by ITS rRNA gene region sequence analysis. A microsporidia species obtained from humans in Türkiye has been characterized for the first time and registered in GenBank. Conclusion: Our epidemiological results show that the prevalence of renal microsporidiosis in kidney transplant recipients is very low. In addition, as a result of the phylogenetic analysis of the detected isolate, it was observed that it was 100% identical to the isolates reported from dogs in Kayseri, Türkiye. This situation provided essential data regarding the zoonotic transmission dynamics of microsporidia.
Assuntos
Encephalitozoon , Encefalitozoonose , Transplante de Rim , Microsporidiose , Filogenia , Humanos , Transplante de Rim/efeitos adversos , Prevalência , Masculino , Adulto , Encefalitozoonose/epidemiologia , Feminino , Encephalitozoon/genética , Encephalitozoon/isolamento & purificação , Criança , Turquia/epidemiologia , Microsporidiose/epidemiologia , Pessoa de Meia-Idade , Adolescente , Adulto Jovem , Reação em Cadeia da Polimerase , Hospedeiro Imunocomprometido , Pré-Escolar , Idoso , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , AnimaisRESUMO
Introduction: Wild rodents can serve as reservoirs or carriers of E. bieneusi, thereby enabling parasite transmission to domestic animals and humans. This study aimed to investigate the prevalence of E. bieneusi in wild rodents from the Inner Mongolian Autonomous Region and Liaoning Province of China. Moreover, to evaluate the potential for zoonotic transmission at the genotype level, a genetic analysis of the isolates was performed. Methods: A total of 486 wild rodents were captured from two provinces in China. Polymerase chain reaction (PCR) was performed to amplify the vertebrate cytochrome b (cytb) gene in the fecal DNA of the rodents to detect their species. The genotype of E. bieneusi was determined via PCR amplification of the internal transcribed spacer (ITS) region of rDNA. The examination of genetic characteristics and zoonotic potential requires the application of similarity and phylogenetic analysis. Results: The infection rates of E. bieneusi in the four identified rodent species were 5.2% for Apodemus agrarius (n = 89), 4.5% for Cricetulus barabensis (n = 96), 11.3% for Mus musculus (n = 106), and 38.5% for Rattus norvegicus (n = 195). Infection was detected at an average rate of 17.4% among 486 rodents. Of the 11 identified genotypes, nine were known: SHR1 (detected in 32 samples), D (30 samples), EbpA (9 samples), PigEbITS7 (8 samples), HNR-IV (6 samples), Type IV (5 samples), HNR-VII (2 samples), HNH7 (1 sample), and HNPL-V (1 sample). Two novel genotypes were also discovered, NMR-I and NMR-II, each comprising one sample. The genotypes were classified into group 1 and group 13 via phylogenetic analysis. Discussion: Based on the initial report, E. bieneusi is highly prevalent and genetically diverse in wild rodents residing in the respective province and region. This indicates that these animals are crucial for the dissemination of E. bieneusi. Zoonotic E. bieneusi-carrying animals present a significant hazard to local inhabitants. Therefore, it is necessary to increase awareness regarding the dangers presented by these rodents and reduce their population to prevent environmental contamination.
Assuntos
Animais Selvagens , Enterocytozoon , Fezes , Genótipo , Especificidade de Hospedeiro , Microsporidiose , Filogenia , Roedores , Zoonoses , Animais , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , Enterocytozoon/classificação , China/epidemiologia , Zoonoses/microbiologia , Zoonoses/transmissão , Microsporidiose/epidemiologia , Microsporidiose/veterinária , Microsporidiose/microbiologia , Roedores/microbiologia , Fezes/microbiologia , Animais Selvagens/microbiologia , Prevalência , Citocromos b/genética , Reservatórios de Doenças/microbiologia , Camundongos , DNA Espaçador Ribossômico/genética , Humanos , Doenças dos Roedores/microbiologia , Doenças dos Roedores/epidemiologia , Reação em Cadeia da Polimerase , DNA Fúngico/genética , RatosRESUMO
Enterocytozoon bieneusi microsporidia are emerging pathogens infecting a wide range of vertebrate and invertebrate hosts, known to have zoonotic features since they infect both wild and domestic animals, and humans. Despite their significance, there is very limited epidemiological data on microsporidia in hedgehogs, especially European hedgehogs (Erinaceus europaeus) and long-eared hedgehogs (Hemiechinus auritus), the former known as synantropic hedgehogs, and the latter suited as pets. As such, the present study aimed to assess the presence of E. bieneusi in hedgehogs from Portugal. For this purpose, fecal samples from 110 hedgehogs of three species-E. europaeus (n = 106), H. auritus (n = 1), and Atelerix albiventris (n = 3)-were collected and tested for E. bieneusi by PCR targeting the internal transcribed spacer region and the flanking small and large subunits of the rRNA. We found an overall occurrence of 22.7% (25/110; 95% confidence interval [CI]: 15.28-31.70), with 22.6% (24/106; 95% [CI]: 15.08-31.79) in E. europaeus, 100% (1/1) in H. auritus, and 0% in A. albiventris. Interestingly, three novel genotypes were identified, all belonging to the potentially zoonotic Group 1. Our findings highlight the importance of hedgehogs as potential reservoirs for E. bieneusi and emphasize the need for further research to understand their role in transmission dynamics and assess the associated risks to public and veterinary health.
Synanthropic hedgehogs were tested for Enterocytozoon bieneusi, the main cause of human microsporidiosis. Results showed 22.7% of hedgehogs were shedding E. bieneusi spores, with three new genotypes from the zoonotic Group 1. Hedgehogs may transmit to humans/animals, warranting more research.
Assuntos
DNA Fúngico , Enterocytozoon , Fezes , Ouriços , Microsporidiose , Ouriços/microbiologia , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , Enterocytozoon/classificação , Animais , Microsporidiose/veterinária , Microsporidiose/epidemiologia , Microsporidiose/microbiologia , Portugal/epidemiologia , Fezes/microbiologia , DNA Fúngico/genética , DNA Espaçador Ribossômico/genética , Reação em Cadeia da Polimerase , Filogenia , Análise de Sequência de DNA , GenótipoRESUMO
Enterocytozoon bieneusi is an obligate intracellular microsporidian parasite with a worldwide distribution. As a zoonotic pathogen, E. bieneusi can infect a wide range of wildlife hosts through the fecal-oral route. Although the feces of flying squirrels (Trogopterus xanthipes) are considered a traditional Chinese medicine (as "faeces trogopterori"), no literature is available on E. bieneusi infection in flying squirrels to date. In this study, a total of 340 fresh flying squirrel fecal specimens from two captive populations were collected in Pingdingshan city, China, to detect the prevalence of E. bieneusi and assess their zoonotic potential. By nested PCR amplification of the ITS gene, six specimens tested positive, with positive samples from each farm, with an overall low infection rate of 1.8%. The ITS sequences revealed three genotypes, including known genotype D and two novel genotypes, HNFS01 and HNFS02. Genotype HNFS01 was the most prevalent (4/6, 66.7%). Phylogenetic analysis showed that all genotypes clustered into zoonotic Group 1, with the novel genotypes clustering into different subgroups. To our knowledge, this is the first report of E. bieneusi infection in flying squirrels, suggesting that flying squirrels could act as a potential reservoir and zoonotic threat for E. bieneusi transmission to humans in China.
Title: Occurrence et génotypage d'Enterocytozoon bieneusi chez les écureuils volants (Trogopterus xanthipes) de Chine. Abstract: Enterocytozoon bieneusi est un parasite microsporidien intracellulaire obligatoire présent dans le monde entier. En tant qu'agent pathogène zoonotique, E. bieneusi peut infecter un large éventail d'hôtes sauvages par la voie fécale-orale. Bien que les excréments d'écureuils volants (Trogopterus xanthipes) soient considérés comme un ingrédient de médecine traditionnelle chinoise (comme « faeces trogopterori ¼), aucune littérature n'est disponible à ce jour sur l'infection par E. bieneusi chez les écureuils volants. Dans cette étude, un total de 340 spécimens fécaux frais d'écureuils volants provenant de deux populations captives ont été collectés dans la ville de Pingdingshan, en Chine, pour détecter la prévalence d'E. bieneusi et évaluer leur potentiel zoonotique. Par amplification PCR nichée du gène ITS, six échantillons se sont révélés positifs, avec des échantillons positifs dans chaque ferme, et un taux d'infection global faible, à 1,8 %. Les séquences ITS ont révélé trois génotypes, dont le génotype D connu et deux nouveaux génotypes, HNFS01 et HNFS02. Le génotype HNFS01 était le plus répandu (4/6, 66,7 %). L'analyse phylogénétique a montré que tous les génotypes se regroupaient dans le groupe zoonotique 1, les nouveaux génotypes se regroupant en différents sous-groupes. À notre connaissance, il s'agit du premier rapport d'infection par E. bieneusi chez des écureuils volants, ce qui suggère que les écureuils volants pourraient agir comme un réservoir potentiel et une menace zoonotique pour la transmission d'E. bieneusi aux humains en Chine.
Assuntos
Enterocytozoon , Fezes , Genótipo , Microsporidiose , Filogenia , Sciuridae , Animais , Sciuridae/microbiologia , Sciuridae/parasitologia , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , Enterocytozoon/classificação , China/epidemiologia , Microsporidiose/veterinária , Microsporidiose/epidemiologia , Microsporidiose/microbiologia , Fezes/microbiologia , Fezes/parasitologia , Prevalência , Zoonoses , Reação em Cadeia da Polimerase/veterinária , DNA Fúngico/genética , Doenças dos Roedores/epidemiologia , Doenças dos Roedores/microbiologia , Doenças dos Roedores/parasitologia , DNA Espaçador Ribossômico/genética , Animais Selvagens/microbiologiaRESUMO
BACKGROUND: Parasites Entamoeba spp., Enterocytozoon bieneusi and Blastocystis are prevalent pathogens causing gastrointestinal illnesses in animals and humans. Consequently, researches on their occurrence, distribution and hosts are crucial for the well-being of both animals and humans. Due to the confined spaces and frequent interaction between animals and humans, animal sanctuaries have emerged as potential reservoirs for these parasites. In this study, the wildlife sanctuary near the Huang Gorge of the Qinling Mountains in northwest China is chosen as an ideal site for parasite distribution research, considering its expansive stocking area and high biodiversity. RESULTS: We collected 191 fecal specimens from 37 distinct wildlife species and extracted genomic DNA. We identified these three parasites by amplifying specific gene regions and analyzed their characteristics and evolutionary relationships. All the parasites exhibited a high overall infection rate, reaching 90.05%. Among them, seven Entamoeba species were identified, accounting for a prevalence of 54.97%, with the highest infection observed in Entamoeba bovis. In total, 11 Enterocytozoon bieneusi genotypes were discovered, representing a prevalence of 35.08%, including three genotypes of human-pathogenic Group 1 and two novel genotypes (SXWZ and SXLG). Additionally, 13 Blastocystis subtypes were detected, showing a prevalence of 74.87% and encompassing eight zoonotic subtypes. All of the above suggests significant possibilities of parasite transmission between animals and humans. CONCLUSIONS: This study investigated the occurrence and prevalence of three intestinal parasites, enhancing our understanding of their genetic diversity and host ranges in northwest China. Furthermore, the distribution of these parasites implies significant potential of zoonotic transmission, underscoring the imperative for ongoing surveillance and implementation of control measures. These efforts are essential to mitigate the risk of zoonotic disease outbreaks originating from wildlife sanctuary.
Assuntos
Animais Selvagens , Blastocystis , Entamoeba , Enterocytozoon , Microsporidiose , Zoonoses , Animais , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , China/epidemiologia , Blastocystis/genética , Blastocystis/classificação , Blastocystis/isolamento & purificação , Animais Selvagens/parasitologia , Zoonoses/parasitologia , Entamoeba/genética , Entamoeba/isolamento & purificação , Entamoeba/classificação , Microsporidiose/veterinária , Microsporidiose/epidemiologia , Filogenia , Fezes/parasitologia , Entamebíase/veterinária , Entamebíase/epidemiologia , Entamebíase/parasitologia , Infecções por Blastocystis/veterinária , Infecções por Blastocystis/epidemiologia , Infecções por Blastocystis/transmissão , Infecções por Blastocystis/parasitologia , Prevalência , Genótipo , HumanosRESUMO
Enterocytozoon bieneusi is a widespread intracellular fungus that can infect both humans and animals, making it a significant zoonotic threat. In the current study, a total of 208 fecal samples were assayed to investigate the prevalence of E. bieneusi in pigs reared in Zhejiang Province, China. Employing polymerase chain reaction (PCR) amplification techniques specifically designed to target the internal transcribed spacer (ITS) region of the small subunit ribosomal RNA (rRNA) gene, the results revealed that 78 samples (37.5â¯%) tested positive for the presence of E. bieneusi. A total of 19 different genotypes of E. bieneusi were detected. Nine of these genotypes were already known: EbpC (n = 36), KIN-1 (n = 10), PigEbITS7 (n = 8), EbpA (n = 6), Henan III (n = 3), PigEbITS5 (n = 2), Henan-IV (n = 1), EbpD (n = 1), and TypeIV (n = 1), and 10 were novel: ZJP-I to ZJP-X (one each). The present investigation revealed that all the nine known genotypes identified in pigs here, have also been previously discovered in humans. Additionally, the novel genotypes of E. bieneusi discovered here were all classified as belonging to Group 1. These findings suggest the potential for cross-species transmission between humans and pigs.
Assuntos
Enterocytozoon , Genótipo , Doenças dos Suínos , Zoonoses , Animais , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , China/epidemiologia , Suínos , Doenças dos Suínos/microbiologia , Doenças dos Suínos/epidemiologia , Zoonoses/microbiologia , Filogenia , Medição de Risco , Fezes/microbiologia , Humanos , Prevalência , Microsporidiose/veterinária , Microsporidiose/epidemiologia , Microsporidiose/microbiologia , Reação em Cadeia da Polimerase/veterinária , DNA Espaçador Ribossômico/genética , DNA Fúngico/genéticaRESUMO
White feces syndrome (WFS) is a multifactorial disease that affects global shrimp production. The diagnostic approach to identify WFS involves traditional and molecular scientific methods by examining histopathology, bioassays, PCR (polymerase chain reaction), and calorimetric estimation. The pathogenesis of WFS is closely associated with Vibrio spp., intestinal microbiota (IM) dysbiosis, and Enterocytozoon hepatopenaei (EHP). It also has caused over 10-15 % loss in the aquaculture industry and is also known to cause retardation, lethargy and slowly leading to high mortality in shrimp farms. Therefore, it is necessary to understand the molecular mechanisms processed under the association of IM dysbiosis, Vibrio spp., and EHP to analyze the impact of disease on the innate immune system of shrimp. However, only very few reviews have described the molecular pathways involved in WFS. Hence, this review aims to elucidate an in-depth analysis of molecular pathways involved in the innate immune system of shrimp and their response to pathogens. The analysis and understanding of the impact of shrimp's innate immune system on WFS would help in developing treatments to prevent the spread of disease, thereby improving the economic condition of shrimp farms worldwide.
Assuntos
Imunidade Inata , Penaeidae , Animais , Penaeidae/imunologia , Penaeidae/genética , Penaeidae/microbiologia , Imunidade Inata/genética , Microbioma Gastrointestinal/imunologia , Vibrio/fisiologia , Disbiose/imunologia , Disbiose/veterinária , Enterocytozoon/genética , Enterocytozoon/imunologia , AquiculturaRESUMO
Enterocytozoon bieneusi is a common cause of human microsporidiosis and can infect a variety of animal hosts worldwide. In Thailand, previous studies have shown that this parasite is common in domestic animals. However, information on the prevalence and genotypes of this parasite in other synanthropic wildlife, including bats, remains limited. Several pathogens have been previously detected in bats, suggesting that bats may serve as a reservoir for this parasite. In this study, a total of 105 bat guano samples were collected from six different sites throughout Thailand. Of these, 16 from Chonburi (eastern), Ratchaburi (western), and Chiang Rai (northern) provinces tested positive for E. bieneusi, representing an overall prevalence of 15.2%. Based on ITS1 sequence analysis, 12 genotypes were identified, including two known genotypes (D and type IV) frequently detected in humans and ten novel potentially zoonotic genotypes (TBAT01-TBAT10), all belonging to zoonotic group 1. Lyle's flying fox (Pteropus lylei), commonly found in Southeast Asia, was identified as the host in one sample that was also positive for E. bieneusi. Network analysis of E. bieneusi sequences detected in this study and those previously reported in Thailand also revealed intraspecific divergence and recent population expansion, possibly due to adaptive evolution associated with host range expansion. Our data revealed, for the first time, multiple E. bieneusi genotypes of zoonotic significance circulating in Thai bats and demonstrated that bat guano fertilizer may be a vehicle for disease transmission.
Assuntos
Quirópteros , Enterocytozoon , Genótipo , Microsporidiose , Filogenia , Quirópteros/parasitologia , Quirópteros/microbiologia , Animais , Tailândia/epidemiologia , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , Enterocytozoon/classificação , Microsporidiose/veterinária , Microsporidiose/epidemiologia , Microsporidiose/microbiologia , Prevalência , Humanos , Análise de Sequência de DNA , Zoonoses/parasitologia , DNA Espaçador Ribossômico/genética , DNA Fúngico/genéticaRESUMO
BACKGROUND: Enterocytozoon bieneusi is the most common species found in humans. Although E. bieneusi has been investigated in humans, genotype profile of E. bieneusi is not known in Türkiye. METHODS: In this study, we screened E. bieneusi in patients (n = 94) with different types of malignant solid tumors by Real Time PCR and then sequenced E. bieneusi positive samples. All cancer patients were undergoing chemotherapy and had diarrhea. Moreover, as control groups, we also screened E. bieneusi in patients with diarrhea (n = 50) and without diarrhea (n = 50). RESULTS: Among all patients analyzed, 33 (17%) were found to be E. bieneusi-positive. As the patients were categorized, the molecular prevalence of E. bieneusi increased to 25.5% among cancer patients with diarrhea. However, the molecular prevalence of E. bieneusi was found to be lower in patients with presenting only diarrhea (8%) and patients without diarrhea (10%). The high molecular prevalence value detected among cancer patients with diarrhea was also statistically significant compared to other patient groups (P = 0.00112 and P = 0.0269). Among the 33 Real Time PCR positive samples, 10 of them were amplified by nested PCR and among these 10 samples, 6 of them were successfully genotyped. The phylogenetic tree showed the presence of D and Type IV which were also identified in stray cats living in Izmir in our previous study. CONCLUSIONS: High molecular prevalence value indicates the importance of screening stool samples of cancer patients with diarrhea for E. bieneusi and genotyping results indicate that D and Type IV are circulating between humans and cats.
Assuntos
Diarreia , Enterocytozoon , Genótipo , Microsporidiose , Neoplasias , Humanos , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , Microsporidiose/microbiologia , Microsporidiose/epidemiologia , Microsporidiose/veterinária , Neoplasias/complicações , Neoplasias/tratamento farmacológico , Masculino , Feminino , Diarreia/microbiologia , Diarreia/epidemiologia , Pessoa de Meia-Idade , Prevalência , Adulto , Idoso , Reação em Cadeia da Polimerase em Tempo Real , Adulto Jovem , Filogenia , Análise de Sequência de DNA , Antineoplásicos , DNA Fúngico/genética , Idoso de 80 Anos ou mais , Fezes/microbiologiaRESUMO
To explore the relationship between the intestinal flora of Exopalaemon Carinicauda and infection by Enterocytozoo Hepatopenaei (EHP), we analyzed the species and richness of gut microbiota in infected individuals in different EHP load groups [i.e., control (C), high load (H), and low load (L)] using gene sequencing after infection. The results showed that the abundance of intestinal flora in the high-load EHP group was significantly lower than that in the healthy group. Based on the UPGMA cluster tree and PCoA analysis, with comparisons to healthy shrimp, the gut microbiota of the EHP high load and low load groups were clustered into one branch, which indicated that EHP infection changed the composition of the gut microbiota of infected shrimps. The heat map analysis of species abundance clustering revealed that the dominant bacteria in the low EHP load group and the control group were beneficial genera such as Lactococcus, Ligilactobacillius, and Bifidobacterium, but the dominant bacteria in the high EHP load group were harmful genera such as Pseudomonas, Photobacterium, and Candidatus hepatincola. The functions of the intestinal flora predicted that most genes related to metabolism were more abundant in healthy shrimp, most genes related to metabolism and the organisms' system were more abundant in the low EHP load group, and most genes related to diseases and environmental information processing were more abundant in the high EHP load group. After separation and purification, the dominant bacteria (Bifidobacterium animalis in healthy shrimp and Lactococcus garvieae in the low EHP load group) and the non-dominant bacteria (Macrococus caseolyticus in the low EHP load group) were obtained. Each of these isolated strains were used together with EHP to infect E. carinicauda, and the results showed that Bifidobacterium animali and Lactococcus garvieae significantly reduced the EHP load in EHP-infected individuals. At the same time, the morphology and structure of the hepatopancreas and intestinal tissue of EHP-infected E. carinicauda were improved. No improvement was seen in tissue that was infected with Macrococus caseolyticus.
Assuntos
Enterocytozoon , Microbioma Gastrointestinal , Palaemonidae , Animais , Palaemonidae/microbiologia , Enterocytozoon/genética , Enterocytozoon/fisiologia , Penaeidae/microbiologiaRESUMO
Enterocytozoon bieneusi is the most common microsporidian species in humans and can affect over 200 animal species. Considering possible increasing risk of human E. bieneusi infection due to close contact with pet dogs and identification of zoonotic E. bieneusi genotypes, 589 fresh fecal specimens of pet dogs were collected from Yunnan Province, China to determine the occurrence of E. bieneusi, characterize dog-derived E. bieneusi isolates, and assess their zoonotic potential at the genotype level. Enterocytozoon bieneusi was identified and genotyped by PCR and sequencing of the internal transcribed spacer (ITS) region of the ribosomal RNA (rRNA) gene. Twenty-nine specimens (4.9%) were positive. A statistical difference was observed in occurrence rates of E. bieneusi in pet dogs among 11 sampling sites by Fisher's exact test. Fifteen genotypes were identified and all of them phylogenetically belonged to zoonotic group 1, including four known genotypes (EbpC, D, Peru 8, and Henan-III) and 11 novel genotypes. Genotype Henan-III was reported in dogs for the first time. The finding of known genotypes found previously in humans and novel genotypes falling into zoonotic group 1 indicates that dogs may play a role in the transmission of E. bieneusi to humans in the investigated areas.
Title: Occurrence et caractérisation génétique d'Enterocytozoon bieneusi chez les chiens de compagnie dans la province du Yunnan, Chine. Abstract: Enterocytozoon bieneusi est l'espèce de microsporidies la plus répandue chez l'homme et peut affecter plus de 200 espèces animales. Compte tenu du risque accru possible d'infection humaine à E. bieneusi en raison d'un contact étroit avec des chiens de compagnie et de l'identification de génotypes zoonotiques d'E. bieneusi, 589 échantillons fécaux frais de chiens de compagnie ont été collectés dans la province du Yunnan, en Chine, pour déterminer la présence d'E. bieneusi, caractériser les isolats obtenus de chiens, et évaluer leur potentiel zoonotique au niveau du génotype. Enterocytozoon bieneusi a été identifié et génotypé par PCR et séquençage de la région d'espacement transcrit interne (ITS) du gène de l'ARN ribosomal (ARNr). Vingt-neuf échantillons (4,9%) étaient positifs. Une différence statistique a été observée dans les taux de présence d'E. bieneusi chez les chiens de compagnie parmi 11 sites d'échantillonnage par le test exact de Fisher. Quinze génotypes ont été identifiés et tous appartenaient phylogénétiquement au groupe zoonotique 1, dont quatre génotypes connus (EbpC, D, Peru 8 et Henan-III) et 11 nouveaux génotypes. Le génotype Henan-III est signalé pour la première fois chez le chien. La découverte de génotypes connus précédemment trouvés chez l'homme et de nouveaux génotypes appartenant au groupe zoonotique 1 indique que les chiens peuvent jouer un rôle dans la transmission d'E. bieneusi aux humains dans les zones étudiées.
Assuntos
Doenças do Cão , Enterocytozoon , Fezes , Genótipo , Microsporidiose , Filogenia , Zoonoses , Cães , Animais , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , Enterocytozoon/classificação , China/epidemiologia , Microsporidiose/veterinária , Microsporidiose/epidemiologia , Microsporidiose/microbiologia , Doenças do Cão/epidemiologia , Doenças do Cão/microbiologia , Doenças do Cão/parasitologia , Fezes/microbiologia , Fezes/parasitologia , Animais de Estimação/microbiologia , DNA Espaçador Ribossômico/genética , DNA Fúngico/genética , Humanos , Reação em Cadeia da Polimerase/veterinária , Análise de Sequência de DNARESUMO
The increasing economic losses associated with growth retardation caused by Enterocytozoon hepatopenaei (EHP), a microsporidian parasite infecting penaeid shrimp, require effective monitoring. The internal transcribed spacer (ITS)-1 region, the non-coding region of ribosomal clusters between 18S and 5.8S rRNA genes, is widely used in phylogenetic studies due to its high variability. In this study, the ITS-1 region sequence (~600-bp) of EHP was first identified, and primers for a polymerase chain reaction (PCR) assay targeting that sequence were designed. A newly developed nested-PCR method successfully detected the EHP in various shrimp (Penaeus vannamei and P. monodon) and related samples, including water and feces collected from Indonesia, Thailand, South Korea, India, and Malaysia. The primers did not cross-react with other hosts and pathogens, and this PCR assay is more sensitive than existing PCR detection methods targeting the small subunit ribosomal RNA (SSU rRNA) and spore wall protein (SWP) genes. Phylogenetic analysis based on the ITS-1 sequences indicated that the Indonesian strain was distinct (86.2% nucleotide sequence identity) from other strains collected from Thailand and South Korea, and also showed the internal diversity among Thailand (N = 7, divided into four branches) and South Korean (N = 5, divided into two branches) samples. The results revealed the ability of the ITS-1 region to determine the genetic diversity of EHP from different geographical origins.
Assuntos
DNA Espaçador Ribossômico , Enterocytozoon , Microsporidiose , Penaeidae , Filogenia , Reação em Cadeia da Polimerase , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , Enterocytozoon/classificação , Penaeidae/microbiologia , Penaeidae/parasitologia , Animais , DNA Espaçador Ribossômico/genética , Reação em Cadeia da Polimerase/métodos , Microsporidiose/microbiologia , Microsporidiose/diagnóstico , DNA Fúngico/genética , Primers do DNA/genética , Fezes/microbiologia , Fezes/parasitologia , Análise de Sequência de DNA , TailândiaRESUMO
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.
Assuntos
Doenças dos Bovinos , Criptosporidiose , Cryptosporidium , Enterocytozoon , Giardia lamblia , Giardíase , Microsporidiose , Animais , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , Bovinos , Cryptosporidium/genética , Cryptosporidium/isolamento & purificação , Cryptosporidium/classificação , Giardia lamblia/genética , Giardia lamblia/isolamento & purificação , Criptosporidiose/epidemiologia , Criptosporidiose/parasitologia , China/epidemiologia , Prevalência , Microsporidiose/veterinária , Microsporidiose/epidemiologia , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/parasitologia , Doenças dos Bovinos/microbiologia , Giardíase/veterinária , Giardíase/epidemiologia , Giardíase/parasitologia , Feminino , Fezes/parasitologia , Fezes/microbiologiaRESUMO
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.
Assuntos
Quirópteros , Criptosporidiose , Cryptosporidium , Enterocytozoon , Fezes , Genótipo , Microsporidiose , Animais , Quirópteros/parasitologia , Quirópteros/microbiologia , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , Enterocytozoon/classificação , Cryptosporidium/genética , Cryptosporidium/classificação , Cryptosporidium/isolamento & purificação , China/epidemiologia , Microsporidiose/veterinária , Microsporidiose/epidemiologia , Microsporidiose/parasitologia , Microsporidiose/microbiologia , Criptosporidiose/parasitologia , Criptosporidiose/epidemiologia , Fezes/parasitologia , Fezes/microbiologia , Variação Genética , Filogenia , Análise de Sequência de DNA , DNA Espaçador Ribossômico/genética , Reação em Cadeia da Polimerase , DNA Fúngico/genética , Repetições de Microssatélites , DNA de Protozoário/genética , Parques RecreativosRESUMO
Microsporidia is a diverse group of obligate, intracellular, and spore-forming parasites that infect a wide range of animals. Among them, Enterocytozoon bieneusi and Encephalitozoon spp. are the most frequently reported species in humans. Limited information is available about the presence and molecular diversity of microsporidian species in the endangered Iberian lynx (Lynx pardinus). Presence of Enterocytozoon bieneusi and Encephalitozoon spp. was investigated by molecular methods in wild and captive Iberian lynxes from Spain. Overall, E. bieneusi was detected in 3.2% (8/251) of the animals examined. None of the samples tested were positive for Encephalitozoon spp. Four known (D, EbfelA, PigEBITS7, and Type IV) and a novel (named as LynxSpEb1) E. bieneusi genotypes were identified. All the genotypes found belonged to the zoonotic Group 1 of E. bieneusi. This study provides the first genotyping data of E. bieneusi in Iberian lynx in Spain. Our result indicate that the Iberian lynx does not seem to play a relevant role in the epidemiology of Encephalitozoon spp., and that this endangered felid is likely acting as spillover host rather than a true reservoir of E. bieneusi. Additional studies should be conducted to assess the impact of this parasite in the health status of the endangered Iberian lynx.