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1.
Rev Bras Parasitol Vet ; 29(2): e017919, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32428181

RESUMO

Cryptosporidium is a zoonotic parasite that causes diarrhea in a broad range of animals, including deer. Little is known about the prevalence and genotype of Cryptosporidium spp. in Père David's deer. In this study, 137 fecal samples from Père David's deer were collected between July 2017 and August 2018 in the Dafeng Reserve and analyzed for Cryptosporidium spp. by nested-PCR based on the small subunit ribosomal RNA (SSU rRNA) gene, followed by sequence analyses to determine the species. The 60 kDa glycoprotein (gp60) gene was used to characterize Cryptosporidium spp. Among 137 samples, 2 (1.46%) were positive for Cryptosporidium spp. according to SSU rRNA gene sequencing results. Both samples belonged to the Cryptosporidium deer genotype, with two nucleotide deletions and one nucleotide substitution. The prevalence data and molecular characterization of this study provide basic knowledge for controlling and preventing Cryptosporidium infections in Père David's deer in this area.


Assuntos
Criptosporidiose/epidemiologia , Cryptosporidium/genética , DNA de Protozoário/genética , Cervos/parasitologia , Epidemiologia Molecular , RNA Ribossômico , Animais , China/epidemiologia , Criptosporidiose/parasitologia , Cryptosporidium/isolamento & purificação , Fezes/parasitologia , Genótipo , Filogenia , Prevalência , Análise de Sequência de DNA
2.
Parasitol Res ; 119(6): 1847-1855, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32350588

RESUMO

Cryptosporidium and Giardia are well-known parasitic protozoans responsible for waterborne and foodborne diarrhoeal diseases. However, data are not available on market vegetables contaminated with Cryptosporidium and Giardia in China. In the present study, 642 different vegetable samples were collected from Xining City street vendors in the Qinghai Province to study the Cryptosporidium and Giardia contamination rates via PCR and sequence analyses. Cryptosporidium spp. and Giardia duodenalis were detected in 16 (2.5%) and 73 (11.4%) samples, respectively. Two species of Cryptosporidium, C. parvum (n = 11) and C. andersoni (n = 5), were identified. G. duodenalis assemblage B was identified in almost all positive samples (n = 72), except one sample that contained G. duodenalis assemblage E. We report on the rate of Cryptosporidium and Giardia contamination in vegetables for the first time from the Qinghai Tibetan Plateau Area (QTPA) in China.


Assuntos
Cryptosporidium/isolamento & purificação , Giardia/isolamento & purificação , Oocistos/isolamento & purificação , Verduras/parasitologia , Animais , China , Cryptosporidium/genética , Contaminação de Alimentos , Giardia/genética , Giardia lamblia/genética , Reação em Cadeia da Polimerase , Tibet
3.
PLoS Negl Trop Dis ; 14(4): e0008195, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32320399

RESUMO

BACKGROUND: Intestinal parasites such as Cryptosporidium spp., Giardia lamblia and Entamoeba histolytica can cause severe diarrhea, especially among children in developing countries. This study aims to determine the frequency of Cryptosporidium spp., Giardia lamblia and Entamoeba histolytica in children with diarrhea and identify risk factors for infection. METHODOLOGY: We conducted a cross-sectional study in children aged 0-168 months hospitalized with diarrhea in three regions of Mozambique, from June 2014 to January 2018. Following consent, caretakers were interviewed and a single stool specimen was collected from each child to diagnose Cryptosporidium spp., G. lamblia and E. histolytica using commercial immune-enzymatic assay (TechLab, Inc, Blacksburg, VA, USA). Anthropometric data were collected from the clinical reports. Multivariable logistic regression models were built to identify risk factors for Cryptosporidium spp. and G. lamblia infection. RESULTS: Twenty-one percent of all specimens (212/1008) presented at least one parasitic infection. Cryptosporidium spp. infection was the most common 12.0% (118/985), followed by G. lamblia 9.7% (95/983) and E. histolytica 2.0% (20/1004). Risk factors for infection by Cryptosporidium spp. were: provenience (children from Nampula province showed the highest risk, OR: 8.176; CI: 1.916-34.894; p-value < 0.01); animal contact (children with animal contact had a protective effect OR: 0.627; CI: 0.398-0.986; p-value < 0.05); underweight (children severely underweight showed a risk of 2.309; CI: 1.310-4.069; p-value < 0.05). Risk factors for infection by G. lamblia were: age (group with highest risk, 60-168 months (OR: 2.322; CI: 1.000-5.393, p-value > 0.05)); and living in a household with five or more members (OR: 2.141; CI: 1.286-3.565, p-value < 0.01). CONCLUSIONS: Parasitic infection is common among children with diarrhea. Routine testing, standard treatment, and assessment for risk exposure of children with diarrhea should be implemented at health facilities in Mozambique.


Assuntos
Criptosporidiose/epidemiologia , Diarreia/parasitologia , Entamebíase/epidemiologia , Giardíase/epidemiologia , Adolescente , Animais , Criança , Pré-Escolar , Estudos Transversais , Cryptosporidium/isolamento & purificação , DNA de Protozoário/análise , Diarreia/epidemiologia , Entamoeba histolytica/isolamento & purificação , Fezes/parasitologia , Feminino , Giardia lamblia/isolamento & purificação , Humanos , Lactente , Recém-Nascido , Modelos Logísticos , Masculino , Moçambique/epidemiologia , Análise Multivariada , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Risco
4.
PLoS Negl Trop Dis ; 14(3): e0008146, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32226011

RESUMO

BACKGROUND: Cryptosporidiosis is an emerging infectious disease of public health significance worldwide. The burden of disease caused by Cryptosporidium varies between and within countries/areas. To have a comprehensive understanding of epidemiological status and characteristics of human Cryptosporidium infection in China since the first report in 1987, a retrospective epidemiological analysis was conducted by presenting differences in the prevalence of Cryptosporidium by province, year, population, living environment and season and possible transmission routes and risk factors as well as genetic characteristics of Cryptosporidium in humans. METHODOLOGY/PRINCIPAL FINDINGS: A systematic search was conducted to obtain epidemiological papers of human Cryptosporidium infection/cryptosporidiosis from PubMed and Chinese databases. Finally, 164 papers were included in our analysis. At least 200,054 people from 27 provinces were involved in investigational studies of Cryptosporidium, with an average prevalence of 2.97%. The prevalence changed slightly over time. Variable prevalences were observed: 0.65-11.15% by province, 1.89-47.79% by population, 1.77-12.87% and 0-3.70% in rural and urban areas, respectively. The prevalence peak occurred in summer or autumn. Indirect person-to-person transmission was documented in one outbreak of cryptosporidiosis in a pediatric hospital. 263 Cryptosporidium isolates were obtained, and seven Cryptosporidium species were identified: C. hominis (48.3%), C. andersoni (22.43%), C. parvum (16.7%), C. meleagridis (8.36%), C. felis (3.04%), C. canis (0.76%) and C. suis (0.38%). CONCLUSIONS/SIGNIFICANCES: This systematic review reflects current epidemiological status and characteristics of Cryptosporidium in humans in China. These data will be helpful to develop efficient control strategies to intervene with and prevent occurrence of human Cryptosporidium infection/cryptosporidiosis in China as well as have a reference effect to other countries. Further studies should focus on addressing a high frequency of C. andersoni in humans and a new challenge with respect to cryptosporidiosis with an increasing population of elderly people and patients with immunosuppressive diseases.


Assuntos
Criptosporidiose/epidemiologia , Cryptosporidium/isolamento & purificação , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , China/epidemiologia , Doenças Transmissíveis Emergentes/epidemiologia , Doenças Transmissíveis Emergentes/transmissão , Criptosporidiose/transmissão , Surtos de Doenças , Transmissão de Doença Infecciosa , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Prevalência , Fatores de Risco , Adulto Jovem
5.
Ann Agric Environ Med ; 27(1): 146-150, 2020 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-32208594

RESUMO

INTRODUCTION AND OBJECTIVE: The problem of occupational biohazards is very important, especially in the field of agriculture and in human and veterinary medicine. The aim of the study was to determine the potential sources of infection in veterinary professionals with selected zoonotic agents, including: Toxoplasma gondii, Giardia duodenalis, Leptospira spp., Cryptosporidium spp. and Coxiella burnetii. MATERIAL AND METHODS: A total of 50 air samples from barns, piggeries and veterinary surgeries were examined for the presence of Leptospira spp. and C. burnetii DNA. Serum samples of 86 pigs and 80 cows were tested for the presence of antibodies to Leptospira spp. and to phase I and II C. burnetii antigens. Serum of 70 cats were tested for the presence of antibodies to T. gondii and 65 samples of cat faeces for the presence of T. gondii oocysts. The presence of G. duodenalis and Cryptosporidium spp. were examined in 50 of dog faeces and 50 of bovine faeces samples. RESULTS: DNA of Leptospira spp. was detected in 2 air samples from the piggeries (4%). C. burnetii DNA was not found in any sample. Anti-Leptospira spp. antibodies were detected in 51 (59.3%) of examined pigs. Neither anti-Leptospira spp. nor anti-C. burnetii antibodies were found among samples of bovine serum. Anti-T. gondii antibodies was found in 52 cat serum samples (74.3%). Among samples of cat faeces, no T. gondii oocysts were detected. In one sample of cattle stool (2%), G. duodenalis was detected and in another (2%) - Cryptosporidium spp. G. duodenalis was detected in 7 samples (14%) and Cryptosporidium spp. in 2 samples (2%) of dog faeces. CONCLUSIONS: The results of this study demonstrate the potential risk of infection with Leptospira spp. in veterinarians working with pigs. Veterinarians could be also be at risk of infection with T. gondii and G. duodenalis.


Assuntos
Microbiologia do Ar , Fezes/microbiologia , Fezes/parasitologia , Exposição Ocupacional/análise , Animais , Anticorpos Antiprotozoários/análise , Gatos , Bovinos , Coxiella burnetii/isolamento & purificação , Criptosporidiose , Cryptosporidium/isolamento & purificação , DNA Bacteriano/análise , DNA de Protozoário , Cães , Giardia lamblia/isolamento & purificação , Giardíase/veterinária , Hospitais Veterinários , Abrigo para Animais , Leptospira/isolamento & purificação , Leptospirose/veterinária , Projetos Piloto , Polônia/epidemiologia , Febre Q/veterinária , Suínos , Toxoplasma/isolamento & purificação , Toxoplasmose Animal , Zoonoses/epidemiologia
6.
Vet Clin North Am Food Anim Pract ; 36(1): 223-238, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32029186

RESUMO

Cryptosporidium and Giardia are ubiquitous protozoan parasites that infect a broad range of vertebrate hosts, including domestic and wild animals as well as humans. Both parasites are of medical and veterinary importance. Infections with Cryptosporidium and Giardia in ruminants are associated with diarrhea outbreaks, mainly in young animals. Ruminants are potential sources of infection for humans because some species of Cryptosporidium and assemblages of Giardia duodenalis have been isolated from both ruminants and humans. Knowledge of these parasites has greatly expanded in the last 2 decades from simple microscopic observations of organisms to the knowledge acquired from molecular tools.


Assuntos
Criptosporidiose/parasitologia , Cryptosporidium/isolamento & purificação , Giardia/isolamento & purificação , Giardíase/parasitologia , Giardíase/veterinária , Animais , Fezes/parasitologia , Humanos , Prevalência , Ruminantes/parasitologia
7.
Exp Parasitol ; 210: 107848, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32004534

RESUMO

Marine bivalves are usually cultivated in shallow, estuarine waters where there is a high concentration of nutrients. Many micro-pollutants, including the protozoan parasites Giardia duodenalis and Cryptosporidium spp., which also occur in such environments, may be concentrated in shellfish tissues during their feeding process. Shellfish can thus be considered as vehicles for foodborne infections, as they are usually consumed lightly cooked or raw. Therefore, the main objective of this study was to investigate the presence of both parasites in Mediterranean mussels, Mytilus galloprovincialis that are cultivated in Thermaikos Gulf, North Greece, which is fed by four rivers that are contaminated with both protozoa. Moreover, the occurrence of these protozoa was monitored in treated wastewaters from 3 treatment plants that discharge into the gulf. In order to identify potential sources of contamination and to estimate the risk for human infection, an attempt was made to genotype Giardia and Cryptosporidium in positive samples. Immunofluorescence was used for detection and molecular techniques were used for both detection and genotyping of the parasites. In total, 120 mussel samples, coming from 10 farms, were examined for the presence of both protozoa over the 6-month farming period. None of them were found positive by immunofluorescence microscopy for the presence of parasites. Only in 3 mussel samples, PCR targeting the GP60 gene detected Cryptosporidium spp. DNA, but sequencing was not successful. Thirteen out of 18 monthly samples collected from the 3 wastewater treatment plants, revealed the presence of Giardia duodenalis cysts belonging to sub-assemblage AII, at relatively low counts (up to 11.2 cysts/L). Cryptosporidium oocysts (up to 0.9 oocysts/L) were also detected in 4 out of 8 samples, although sequencing was not successful at any of the target genes. At the studied location and under the sampling conditions described, mussels tested were not found to be harboring Giardia cysts and the presence of Cryptosporidium was found only in few cases (by PCR detection only). Our results suggest that the likelihood that mussels from these locations act as vehicles of human infection for Giardia and Cryptosporidium seems low.


Assuntos
Bivalves/parasitologia , Cryptosporidium/isolamento & purificação , Doenças Transmitidas por Alimentos/parasitologia , Giardia/isolamento & purificação , Animais , Criptosporidiose/transmissão , Cryptosporidium/genética , DNA de Protozoário/isolamento & purificação , Giardia/genética , Giardíase/transmissão , Grécia , Humanos , Oocistos/isolamento & purificação , Reação em Cadeia da Polimerase , Rios/parasitologia , Águas Residuárias/parasitologia
8.
Parasitol Res ; 119(3): 947-956, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31950252

RESUMO

Nowadays, snakes established as domestic exotic pets, harboring numerous (zoonotic) gastrointestinal parasites. In this parasitological survey, we used direct saline fecal smears (DSFS) to examine 586 stool samples from 71 different snake species either kept as pets in households or in zoological gardens in Germany. In addition to DSFS, carbol-fuchsin-fecal smears (n = 296), coproantigen ELISA tests (n = 98), and immunofluorescence assays (IFA; n = 77) for the detection reptile Cryptosporidium infections were conducted. Complete dissections of deceased snakes (n = 63) were also performed in order to gain data on endoparasite species burdens affecting domestic snakes. Overall, examined fecal samples contained 20 different parasite taxa: Ancylostomatid Kalicephalus spp. were the most prevalent nematode species (3.3%), followed by Strongyloides/Rhabdias (2.6%), flagellated protozoan trophozoites (e. g., Proteromonadida, Reteromonadida) (2.3%), Monocercomonas spp. (1.9%), Entamoeba spp. (1.4%), unsporulated coccidian oocysts (1.4%), Kapsulotaenia spp. (0.9%), Capillaria spp. (0.7%), indet. trematodes (0.5%), pentastomids (0.5%), spirurids (0.4%), Eimeria spp. (0.4%), ascarids (0.4%), Blastocystis sp. (0.2%), heterakids (0.2%), cestodes (Proteocephalidae) (0.2%), Plagiorchis spp. (0.2%), Cryptosporidium spp. (0.2%), Caryospora epicratesi (0.2%), and Sarcocystis spp. (0.2%). For Cryptosporidium, four carbol-fuchsin-stained smears (1.4%), 12 (12.2%) coproantigen ELISA-examined samples and 5.2% of examined samples were diagnosed with IFA. Fourteen (22.2%) of dissected snakes showed infections with various pathogenic nematode genera and 8 of them (12.7%) died due to protozoan parasitic infections. High prevalences of intestinal protozoan parasites resulting in severe pathological findings observed in dissected snakes call for more detailed investigations on gastrointestinal parasites.


Assuntos
Criptosporidiose/parasitologia , Enteropatias Parasitárias/parasitologia , Serpentes/parasitologia , Strongyloides/classificação , Strongyloides/isolamento & purificação , Estrongiloidíase/veterinária , Animais , Cryptosporidium/isolamento & purificação , Entamoeba/isolamento & purificação , Fezes/parasitologia , Jardins , Alemanha/epidemiologia , Oocistos/isolamento & purificação , Rhabdiasoidea/classificação , Rhabdiasoidea/isolamento & purificação
9.
BMC Infect Dis ; 20(1): 16, 2020 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-31910816

RESUMO

BACKGROUND: Cryptosporidium is a genus of common intestinal protozoa, members of which cause diarrhea in a wide variety of hosts. Previous studies on Cryptosporidium in China have mainly focused on diarrhea sufferers, children, and immunodeficient individuals such as HIV/AIDS patients. However, the epidemiological characteristics of Cryptosporidium in the population in rural areas remain unclear. Herein, we investigated the prevalence of, and risk factors for, Cryptosporidium in rural areas of Binyang County, Guangxi Zhuang Autonomous Region, China, and genetically characterized the Cryptosporidium isolates we obtained. METHODS: From August to December 2016, two villages in Binyang County, Guangxi, were sampled using a random cluster sampling method. Fresh fecal samples were collected from all eligible residents (residence time > 6 months). Molecular characterization of Cryptosporidium was carried out based on its SSU rRNA, gp60, actin and hsp70 gene sequences. Fisher's exact test were conducted to assess the risk factors for Cryptosporidium infection. RESULTS: A total of 400 fecal samples were collected from 195 males (48.8%) and 205 females (51.2%). Two samples (0.5%) were positive for Cryptosporidium and were identified as C. viatorum and C. occultus respectively. Moreover, a new C. viatorum subtype XVaA3h was identified based on the sequence of the gp 60 gene. CONCLUSIONS: To our knowledge, this is the first report of C. viatorum and C. occultus infections in humans in China and of C. viatorum subtype XVaA3h. The findings provide important information on the prevalence of Cryptosporidium in the Chinese population, and expand the range of Cryptosporidium species known to infect people in China.


Assuntos
Sequência de Bases/genética , Criptosporidiose/epidemiologia , Cryptosporidium/classificação , Cryptosporidium/genética , DNA de Protozoário/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Criança , Pré-Escolar , China/epidemiologia , Estudos Transversais , Criptosporidiose/parasitologia , Cryptosporidium/isolamento & purificação , Diarreia/parasitologia , Fezes/parasitologia , Feminino , Genótipo , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Filogenia , Prevalência , Fatores de Risco , Inquéritos e Questionários , Adulto Jovem
10.
Parasitol Res ; 119(1): 105-114, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31773309

RESUMO

The protistan pathogens Cryptosporidium and Enterocytozoon bieneusi can cause significant intestinal diseases in animals and humans. However, limited information is available regarding prevalence and molecular characterization of Cryptosporidium and E. bieneusi in ruminants in Northern China. In this study, the overall prevalence of Cryptosporidium and E. bieneusi was 19.3% (62/321) and 28.97% (93/321) in dairy calves and 1.10% (9/818) and 13.57% (111/818) in sika deer (Cervus nippon) in four provinces in Northern China, respectively. The prevalence of Cryptosporidium and E. bieneusi in different factor groups was various. Five Cryptosporidium species/genotypes were identified, of which C. parvum, C. ryanae, C. bovis, and C. andersoni were only found in dairy calves, and only Cryptosporidium deer genotype was found in sika deer. Moreover, J, I, and BEB4 ITS genotypes of E. bieneusi were found in dairy calves, and six known genotypes (JLD-III, JLD-IX, JLD-VII, EbpC, BEB6, and I) and ten novel genotypes (namely LND-I and JLD-XV to JLD-XXIII) were found in sika deer in this study. Cryptosporidium parvum and E. bieneusi genotype J were identified as the predominant species/genotypes in dairy calves, whereas the predominance of Cryptosporidium spp. and E. bieneusi in sika deer was Cryptosporidium deer genotype and BEB6, respectively. The present study reported the prevalence and genotypes of Cryptosporidium and E. bieneusi in dairy calves and sika deer in four provinces in northern China. The present findings also suggest that investigated dairy calves and sika deer may play an important role in the transmission of E. bieneusi and Cryptosporidium to humans and other animals, and also in an effort to better understand the epidemiology of these enteric pathogens in China.


Assuntos
Criptosporidiose/epidemiologia , Cryptosporidium/genética , Enterocytozoon/genética , Microsporidiose/epidemiologia , Microsporidiose/veterinária , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/parasitologia , China/epidemiologia , Cryptosporidium/isolamento & purificação , Cervos/parasitologia , Enterocytozoon/isolamento & purificação , Fezes/parasitologia , Feminino , Genótipo , Humanos , Masculino , Prevalência
11.
Sci Total Environ ; 700: 134447, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31677419

RESUMO

Identification of Cryptosporidium oocyst is essential in ensuring water quality fit for human use, consumption, and recreation. This communication proposes the supplemental analysis of substrate-associated biofilms, in particular, freshwater sponges in improving case finding of waterborne-protozoan pathogens (WBPP) in environmental aquatic samples. In this study, a small portion of a mature freshwater sponge under the Genus Spongilla was subjected to microscopic and molecular analysis to identify the presence of Cryptosporidium. Microscopic screening with modified Kinyoun's staining (MK) and microscopic confirmation using direct antibody fluorescent testing (IFT) returned with Cryptosporidium spp. positive findings. Molecular investigation resulted in the confirmation of Cryptosporidium hominis upon sequencing of PCR products and phylogenetic analysis. This is the first report of a pathogenic protozoan, C. hominis isolated from a freshwater sponge. The results of this study provide evidence of the value of expanding water quality assessment strategies to the analysis of substrate-associated biofilms and sponges in improving case finding of WBPP in natural aquatic environments.


Assuntos
Água Doce/microbiologia , Poríferos/parasitologia , Animais , Cryptosporidium/isolamento & purificação , Oocistos , Filogenia , Reação em Cadeia da Polimerase
12.
Exp Parasitol ; 209: 107824, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31870927

RESUMO

Giardiasis and cryptosporidiosis are recognized by the WHO as important emerging diseases of the 21st century. Symptoms are similar and include diarrhoea and vomiting, which may be severe, even life-threatening, for the immunocompromised and children under five years of age. Between 2013 and 2017, the Institute for Public Health in Serbia recorded 10 waterborne epidemics that manifested as gastrointestinal disease. Routine testing for enteropathogenic bacteria and viruses did not identify the aetiological agents of these outbreaks. As water is not examined for the presence of protozoa in Serbia, we performed a pilot study to analyse samples from four major rivers and their tributaries using a newly implemented methodology for detection of Giardia and Cryptosporidium, based on the ISO 15553:2006 standard. Using immunofluorescence microscopy, Giardia was detected in 10 out of the 31 samples, Cryptosporidium in five, while two samples were positive for both. Presence of G. duodenalis gDNA was confirmed by amplification of the ß-giardin gene in eight samples, of which one and two, respectively, were identified by RFLP as potentially zoonotic assemblages A and B. The results suggest that surface water in Serbia may be a potential source of infection and call for more in-depth studies using sophisticated molecular tools.


Assuntos
Cryptosporidium/isolamento & purificação , Giardia/isolamento & purificação , Rios/parasitologia , Animais , Cryptosporidium/genética , Proteínas do Citoesqueleto/genética , DNA de Protozoário/química , DNA de Protozoário/isolamento & purificação , Técnicas de Genotipagem , Giardia/classificação , Giardia/genética , Humanos , Complexo Mediador/genética , Microscopia de Fluorescência , Projetos Piloto , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Proteínas de Protozoários/genética , RNA Ribossômico/genética , Sérvia
13.
Artigo em Inglês | MEDLINE | ID: mdl-31817316

RESUMO

This study quantified the association of rodent fruit damage and the microbiological quality of irrigation water on the risk of microbiological contamination of strawberries collected from 18 U-pick farms across five different districts in the Guangzhou metropolitan region of southern China. Fifty-four composite strawberries samples, with or without evidence of rodent or avian foraging damage (i.e., bitten), along with 16 irrigation water samples, were collected during the spring of 2014 and winter of 2015 from our cohort of 18 farms. Composite strawberry samples and irrigation water were analyzed for total coliforms, E. coli, Salmonella, E. coli O157, Giardia, and Cryptosporidium. Total coliforms and E. coli were detected in 100% and ~90% of irrigation water samples, respectively. In contrast, Cryptosporidium was detected in only two water samples, while Salmonella, E. coli O157, and Giardia were not detected in any water samples. Strawberries with signs of being bitten by wildlife had significantly higher concentrations of total coliforms and E. coli, compared to strawberries with no physical evidence of rodent damage (p < 0.001). Similarly, Cryptosporidium was detected in 7/18 (39%) of bitten, 4/18 (22%) of edge, and 5/18 (28%) of central strawberry samples, respectively. Concentration of E. coli on strawberries (p < 0.001), air temperature (p = 0.025), and presence of Cryptosporidium in irrigation water (p < 0.001) were all associated with the risk of Cryptosporidium contamination on strawberries. Salmonella and Giardia were detected in <4% strawberry samples and E. coli O157 was not detected in any samples. These results indicate the potential food safety and public health risks of consuming unwashed strawberries from U-pick farms, and the need for improved rodent biosecurity of U-pick strawberry fields and enhanced microbiological quality of irrigation water used at these facilities.


Assuntos
Irrigação Agrícola/normas , Fazendas , Contaminação de Alimentos/análise , Fragaria/microbiologia , Abastecimento de Água/normas , Animais , Aves , China , Cryptosporidium/isolamento & purificação , Enterobacteriaceae/isolamento & purificação , Inocuidade dos Alimentos , Giardia/isolamento & purificação , Roedores
14.
Parasitol Res ; 118(12): 3409-3418, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31729572

RESUMO

The arctic fox (Vulpes lagopus), an apex predator with an omnipresent distribution in the Arctic, is a potential source of intestinal parasites that may endanger people and pet animals such as dogs, thus posing a health risk. Non-invasive methods, such as coprology, are often the only option when studying wildlife parasitic fauna. However, the detection and identification of parasites are significantly enhanced when used in combination with methods of molecular biology. Using both approaches, we identified unicellular and multicellular parasites in faeces of arctic foxes and carcasses of sibling voles (Microtus levis) in Svalbard, where molecular methods are used for the first time. Six new species were detected in the arctic fox in Svalbard, Eucoleus aerophilus, Uncinaria stenocephala, Toxocara canis, Trichuris vulpis, Eimeria spp., and Enterocytozoon bieneusi, the latter never found in the arctic fox species before. In addition, only one parasite was found in the sibling vole in Svalbard, the Cryptosporidium alticolis, which has never been detected in Svalbard before.


Assuntos
Arvicolinae/parasitologia , Raposas/parasitologia , Enteropatias Parasitárias/veterinária , Ancylostomatoidea/isolamento & purificação , Animais , Animais Selvagens/parasitologia , Regiões Árticas , Cryptosporidium/isolamento & purificação , Cães , Eimeria/isolamento & purificação , Fezes/parasitologia , Helmintos/isolamento & purificação , Nematoides/isolamento & purificação , Svalbard , Toxocara canis/isolamento & purificação , Trichuris/isolamento & purificação
15.
Infect Dis Poverty ; 8(1): 96, 2019 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-31775877

RESUMO

BACKGROUND: There is significant need for accurate diagnostic tools for Cryptosporidium spp. and Giardia duodenalis infections in resource limited countries where diarrhoeal disease caused by these parasites is often prevalent. The present study assessed the diagnostic performance of three commercially available rapid diagnostic tests (RDTs) based on faecal-antigen detection for Cryptosporidium spp. and/or G. duodenalis infections in stool samples of children admitted with severe acute malnutrition (SAM) and diarrhoea. An established multiplex PCR was used as reference test. METHODS: Stool samples from children with SAM and diarrhoea enrolled in a randomized controlled trial (registered at clinicaltrials.gov/ct2/show/NCT02246296) in Malawi (n = 175) and Kenya (n = 120) between December 2014 and December 2015 were analysed by a multiplex PCR for the presence of Cryptosporidium spp., G. duodenalis or Entamoeba histolytica parasite DNA. Cryptosporidium-positive samples were species typed using restriction fragment length polymorphism analysis. A sub-sample of the stool specimens (n = 236) was used for testing with three different RDTs. Diagnostic accuracy of the tests under evaluation was assessed using the results of PCR as reference standard using MedCalc software. Pearson Chi-square test and Fisher's exact test were used to determine (significant) difference between the number of cryptosporidiosis or giardiasis cases found by PCR in Malawi and Kenya. The overall diagnostic accuracy of each RDT was calculated by plotting a receiver operating characteristic (ROC) curve for each test and to determine the area under the curve (AUC) using SPSS8 software. RESULTS: Prevalence of Cryptosporidium spp. by PCR was 20.0 and 21.7% in Malawi and Kenya respectively, mostly C. hominis. G. duodenalis prevalence was 23.4 and 5.8% in Malawi and Kenya respectively. E. histolytica was not detected by PCR. RDT testing followed the same pattern of prevalence. RDT sensitivities ranged for cryptosporidiosis from 42.9 to 76.9% and for G. duodenalis from 48.2 to 85.7%. RDT specificities ranged from 88.4 to 100% for Cryptosporidium spp. and from 91.2 to 99.2% for G. duodenalis infections. Based on the estimated area under the curve (AUC) values, all tests under evaluation had an acceptable overall diagnostic accuracy (> 0.7), with the exception of one RDT for Cryptosporidium spp. in Malawi. CONCLUSIONS: All three RDTs for Cryptosporidium spp. and Giardia duodenalis evaluated in this study have a moderate sensitivity, but sufficient specificity. The main value of the RDTs is within their rapidness and their usefulness as screening assays in surveys for diarrhoea.


Assuntos
Criptosporidiose/epidemiologia , Cryptosporidium/isolamento & purificação , Testes Diagnósticos de Rotina/métodos , Giardia lamblia/isolamento & purificação , Giardíase/epidemiologia , Pré-Escolar , Criptosporidiose/diagnóstico , Testes Diagnósticos de Rotina/instrumentação , Fezes/parasitologia , Giardíase/diagnóstico , Humanos , Lactente , Recém-Nascido , Quênia/epidemiologia , Malaui/epidemiologia , Prevalência , Sensibilidade e Especificidade , Desnutrição Aguda Grave/etiologia
16.
BMC Vet Res ; 15(1): 417, 2019 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-31752852

RESUMO

BACKGROUND: With worldwide distribution and importance for veterinary medicine, Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi have been found in a wide variety of vertebrate hosts. At present, few available molecular data can be used to understand the features of genetic diversity of these pathogens in areas without or less intensive farming. Dominated by grazing, Tibet is a separate geographic unit in China and yaks are in frequent contact with local herdsmen and necessary for their daily life. Therefore, to investigate the distribution of these pathogens in yaks of Tibet, 577 fecal specimens were screened using nested PCR for the presence and genotypes of the three intestinal pathogens. RESULTS: The overall prevalence of Cryptosporidium spp., G. duodenalis, and E. bieneusi were 1.4% (8/577), 1.7% (10/577), and 5.0% (29/577), respectively. Cryptosporidium andersoni (n = 7) and Cryptosporidium bovis (n = 1) were detected by sequence analysis of the SSU rRNA gene. Genotyping at the SSU rRNA and triosephosphate isomerase genes suggested that all G. duodenalis positive specimens belonged to assemblage E. Sequence analysis of the internal transcribed spacer gene identified six known E. bieneusi genotypes: BEB4 (n = 11), I (n = 6), D (n = 5), J (n = 2), CHC8 (n = 1), and BEB6 (n = 1). One subtype (A5,A4,A2,A1) for C. andersoni and three multilocus genotypes for E. bieneusi were identified by multilocus sequence typing. CONCLUSIONS: We report for the first time the status of three enteric pathogens infection simultaneously for grazing yaks in Tibet. Yaks in our study are likely to impose a low zoonotic risk for humans. The molecular epidemiology data add to our knowledge of the characteristics of distribution and transmission for these pathogens in Tibet and their zoonotic potential and public health significance.


Assuntos
Doenças dos Bovinos/parasitologia , Cryptosporidium/isolamento & purificação , Enterocytozoon/isolamento & purificação , Giardia lamblia/isolamento & purificação , Giardíase/veterinária , Microsporidiose/veterinária , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Criptosporidiose/epidemiologia , Criptosporidiose/parasitologia , Cryptosporidium/genética , Enterocytozoon/genética , Giardia lamblia/genética , Giardíase/epidemiologia , Giardíase/parasitologia , Microsporidiose/epidemiologia , Microsporidiose/microbiologia , Filogenia , Especificidade da Espécie , Tibet/epidemiologia
17.
Parasitol Res ; 118(12): 3555-3559, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31722067

RESUMO

The aim of this study was to survey the Cryptosporidium species in peafowls (Pavo cristatus) in Henan Province, China. A total of 143 fecal specimens collected from a breeding farm were tested for Cryptosporidium by nested PCR targeting the small subunit rRNA (SSU rRNA), 70-kDa heat shock protein (HSP70), and actin genes of Cryptosporidium followed by sequence analysis. Only one isolate from an asymptomatic host was obtained, and the isolate differed from a new C. xiaoi-like genotype by one nucleotide and from C. xiaoi or C. bovis at the SSU rRNA locus by six nucleotides. Likewise, the actin gene shared 99% identity with the C. xiaoi-like genotype, accompanied by four nucleotide mutations. A complete sequence of the HSP70 gene was obtained, and exhibited 96% similarity with that from C. xiaoi and differed by one nucleotide from that with the C. xiaoi-like genotype. Phylogenetic analysis of the current isolate revealed genetic relatedness to the C. xiaoi-like genotype and distinction from C. xiaoi and C. bovis. Therefore, our results provided the first documentation of avian infection with a C. xiaoi-like genotype in China and further insight into the diversity of Cryptosporidium spp. in avians.


Assuntos
Doenças das Aves/parasitologia , Criptosporidiose/parasitologia , Cryptosporidium/isolamento & purificação , Animais , Doenças das Aves/epidemiologia , China/epidemiologia , Criptosporidiose/epidemiologia , Cryptosporidium/classificação , Cryptosporidium/genética , DNA de Protozoário/genética , Fezes/parasitologia , Galliformes/parasitologia , Genótipo , Filogenia , Reação em Cadeia da Polimerase/veterinária , RNA Ribossômico/genética
18.
Parasit Vectors ; 12(1): 543, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31730024

RESUMO

BACKGROUND: Cryptosporidium spp. and Giardia duodenalis are major intestinal pathogens that can cause diarrheal diseases in humans, especially children. Enterocytozoon bieneusi is another parasite which can cause gastrointestinal tract disorders, with diarrhea being the main clinical symptom. However, few genetic studies of these parasites in pediatric inpatients in China have been published. METHODS: To assess the genetic characteristics and epidemiological status of these parasites, a total of 2284 fecal samples were collected from children in the pediatric departments of three hospitals in Zhengzhou, central China, and screened for these protozoans with PCR, based on the small subunit ribosomal RNA (SSU rRNA) genes of Cryptosporidium spp. and G. duodenalis and the internal transcribed spacer (ITS) of E. bieneusi. RESULTS: Six (0.26%), 14 (0.61%), and 27 (1.18%) of the samples were positive for Cryptosporidium spp., G. duodenalis and E. bieneusi, respectively. Of the 12 successfully sequenced G. duodenalis isolates, four were identified as assemblage A and eight as assemblage B. In subtype and multilocus genotype (MLG) analyses, C. parvum IIdA19G1 (n = 4) and two novel G. duodenalis MLGs belonging to subassemblage AII (n = 3) and BIV (n = 5) were successfully identified. The E. bieneusi isolates included genotypes D (n = 17), J (n = 2), PigEBITS7 (n = 1), BEB6 (n = 1), and CM8 (n = 1). This is the first report of C. parvum subtype IIdA19G1 in HIV-negative children and E. bieneusi genotype CM8 in humans. CONCLUSIONS: The dominance of zoonotic C. parvum subtype IIdA19G1 indicates that this parasite is turning into zoonotic origin from human-to-human transmission. The phylogenetic analysis also revealed the zoonotic origins and anthroponotic transmission potential of G. duodenalis and E. bieneusi, suggesting more efforts must be made to minimize the threat these pathogens pose to public health.


Assuntos
Cryptosporidium/classificação , Cryptosporidium/genética , Enterocytozoon/classificação , Enterocytozoon/genética , Fezes/parasitologia , Giardia lamblia/classificação , Giardia lamblia/genética , Adolescente , Criança , Criança Hospitalizada , Pré-Escolar , China , Análise por Conglomerados , Criptosporidiose/parasitologia , Cryptosporidium/isolamento & purificação , DNA de Protozoário/química , DNA de Protozoário/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Transmissão de Doença Infecciosa , Enterocytozoon/isolamento & purificação , Feminino , Variação Genética , Genótipo , Giardia lamblia/isolamento & purificação , Giardíase/parasitologia , Hospitais Pediátricos , Humanos , Lactente , Masculino , Microsporidiose/parasitologia , Filogenia , RNA Ribossômico 18S/genética , Análise de Sequência de DNA
19.
Parasit Vectors ; 12(1): 502, 2019 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-31661007

RESUMO

BACKGROUND: Cryptosporidium viatorum is a minor Cryptosporidium pathogen in humans. Currently, there is limited information regarding the prevalence and genotypes of C. viatorum in animals in China. METHODS: In this study, 228 faecal samples were collected from two wild rat species (Leopoldamys edwardsi and Berylmys bowersi) in Chongqing Municipality and Guangdong Province, China. These specimens were analyzed for C. viatorum and then subtyped it using PCR and sequence analysis of the small subunit ribosomal RNA (SSU rRNA) and 60-kilodalton glycoprotein (gp60) genes, respectively. RESULTS: A total of 25 (11.0%) faecal samples were tested positive for C. viatorum by SSU rRNA assay. Of these samples, 4 (3.6%) came from L. edwardsi and 21 (18.0%) from B. bowersi. Of the 25 C. viatorum-positive samples, 17 were successfully amplified at the gp60 gene locus, which represented four subtypes belonging to two subtype families, including XVa (XVaA6, XVaA3g, XVaA3h) and XVc (XVcA2G1). Phylogenetic analysis based on the gp60 amino acid sequences indicated that all of the C. viatorum isolates grouped together, supporting the conclusion that C. viatorum from the wild rats represent two subtype families. CONCLUSIONS: These results indicate an occurrence of C. viatorum XVa subtype family from rats which is genetically identical to those found in humans. Our findings suggest that wild rats may be a potential source of human cryptosporidiosis.


Assuntos
Criptosporidiose/parasitologia , Cryptosporidium/isolamento & purificação , Murinae/parasitologia , Doenças dos Roedores/parasitologia , Sequência de Aminoácidos , Animais , China/epidemiologia , Criptosporidiose/epidemiologia , Criptosporidiose/transmissão , Cryptosporidium/classificação , Cryptosporidium/genética , DNA de Protozoário/genética , DNA de Protozoário/isolamento & purificação , Fezes/parasitologia , Humanos , Filogenia , Reação em Cadeia da Polimerase/veterinária , Prevalência , RNA Ribossômico/química , Doenças dos Roedores/epidemiologia , Doenças dos Roedores/transmissão , Sialoglicoproteínas/química , Sialoglicoproteínas/genética
20.
PLoS Negl Trop Dis ; 13(10): e0007750, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31661498

RESUMO

BACKGROUND: The World Health Organization WHO has estimated that in developed countries, up to 30% of the population may suffer from foodborne diseases each year, and that in developing countries up to 2 million deaths per annum can be attributed to cryptosporidiosis. Reports have already emphasized the role of immigrants in outbreaks of parasitic diseases especially those working in food processing industries. METHODOLOGY/PRINCIPAL FINDINGS: Herein we assessed Cryptosporidium spp. infections among immigrants in Qatar with a special focus on food handlers and housemaids. The overall prevalence of Cryptosporidium spp. by q-PCR among 839 asymptomatic subjects was 4.5%. Based on the Gp60 gene, the majority of isolates were identified as C. parvum subtype IIdA20G1b. The positive sample for C. hominis was subtyped as IeA12G3T3. Seven mixed infections were also identified (four C. parvum + C. hominis, and three C. parvum + C. meleagridis). The prevalence of Cryptosporidium spp. did not differ significantly between the sexes or age classes but varied significantly between subjects affiliated to different religions with the lowest prevalence among the Muslims. Multifactorial analysis retained also marked significance with education, income, and a house contents index. CONCLUSIONS/SIGNIFICANCE: Our results contribute to a better understanding of the epidemiology of cryptosporidiosis and the risk factors associated with the likelihood of carrying this infection among immigrant workers from developing countries.


Assuntos
Criptosporidiose/epidemiologia , Cryptosporidium/isolamento & purificação , Migrantes , Adolescente , Adulto , Criptosporidiose/diagnóstico , Cryptosporidium/genética , DNA de Protozoário/genética , Surtos de Doenças , Feminino , Genes de Protozoários , Genótipo , Técnicas de Genotipagem , Humanos , Masculino , Pessoa de Meia-Idade , Tipagem de Sequências Multilocus , Patologia Molecular , Prevalência , Catar/epidemiologia , Fatores de Risco , Adulto Jovem
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