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1.
MMWR Morb Mortal Wkly Rep ; 70(20): 733-738, 2021 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-34014907

RESUMO

Outbreaks associated with treated recreational water can be caused by pathogens or chemicals in aquatic venues such as pools, hot tubs, water playgrounds, or other artificially constructed structures that are intended for recreational or therapeutic purposes. For the pseriod 2015-2019, public health officials from 36 states and the District of Columbia (DC) voluntarily reported 208 outbreaks associated with treated recreational water. Almost all (199; 96%) of the outbreaks were associated with public (nonbackyard) pools, hot tubs, or water playgrounds. These outbreaks resulted in at least 3,646 cases of illness, 286 hospitalizations, and 13 deaths. Among the 155 (75%) outbreaks with a confirmed infectious etiology, 76 (49%) were caused by Cryptosporidium (which causes cryptosporidiosis, a gastrointestinal illness) and 65 (42%) by Legionella (which causes Legionnaires' disease, a severe pneumonia, and Pontiac fever, a milder illness with flu-like symptoms). Cryptosporidium accounted for 2,492 (84%) of 2,953 cases resulting from the 155 outbreaks with a confirmed etiology. All 13 deaths occurred in persons affected by a Legionnaires' disease outbreak. Among the 208 outbreaks, 71 (34%) were associated with a hotel (i.e., hotel, motel, lodge, or inn) or a resort, and 107 (51%) started during June-August. Implementing recommendations in CDC's Model Aquatic Health Code (MAHC) (1) can help prevent outbreaks associated with treated recreational water in public aquatic venues.


Assuntos
Doenças Transmissíveis/epidemiologia , Surtos de Doenças/estatística & dados numéricos , Recreação , Purificação da Água/estatística & dados numéricos , Banhos/efeitos adversos , Doenças Transmissíveis/etiologia , Cryptosporidium/isolamento & purificação , Estâncias para Tratamento de Saúde/estatística & dados numéricos , Humanos , Legionella/isolamento & purificação , Piscinas/estatística & dados numéricos , Estados Unidos/epidemiologia , Microbiologia da Água
2.
Exp Parasitol ; 225: 108113, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33992605

RESUMO

Cryptosporidiosis remains the leading protozoan induced cause of diarrhoea-associated mortality worldwide. Cryptosporidium hominis, the anthroponotically transmitted species within the Cryptosporidium genus, contributes significantly to the global burden of infection, accounting for the majority of clinical cases in many countries. This study applied high resolution melting analysis, a post-real-time PCR application, to the differentiation of six globally prevalent C. hominisgp60-subtypes. This novel method targeted three microsatellite, tandem repeat containing genetic markers, gp60, the genetic marker upon which current Cryptosporidium subtype nomenclature is based, MSB, and MSE, by which to differentiate between C. hominis isolates. This multi-locus approach successfully differentiated between all six C. hominisgp60-subtypes studied, some of which, such as IbA10G2, are known to exhibit global ubiquity. Thus, this method has the potential to be universally employed as a sensitive, cost effective and highly reproducible means to rapidly differentiate between C. hominisgp60-subtypes. Such a method would be of particular utility in epidemiological studies and outbreak scenarios, providing cost effective, clinically accessible alternative to DNA sequencing. The success of this preliminary study also supports further analysis of an expanded C. hominisgp60-subtype range and the potential expansion of the multi-locus panel in order to improve the discriminatory power of this approach.


Assuntos
Cryptosporidium/genética , Parasitologia/métodos , Criptosporidiose/parasitologia , Cryptosporidium/classificação , Cryptosporidium/isolamento & purificação , DNA de Protozoário/genética , Fezes/parasitologia , Marcadores Genéticos , Genótipo , Humanos , Tipagem de Sequências Multilocus , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Análise de Sequência de DNA
3.
BMC Infect Dis ; 21(1): 349, 2021 Apr 14.
Artigo em Inglês | MEDLINE | ID: mdl-33853528

RESUMO

BACKGROUND: Cryptosporidiosis is a disease caused by infection with an intestinal coccidian parasite Cryptosporidium. Cryptosporidium species are the second leading cause of diarrheal disease and death in children in developing countries. Until now, no data have been available or published on its prevalence among children with diarrhea in Sudan. Therefore, this paper was designed to determine the prevalence rate of Cryptosporidium among children with diarrhea under 5 years who were admitted to Kosti Teaching Hospital. METHODS: A hospital-based cross-sectional study including children under 5 years old admitted to the pediatric section of the hospital between September 2020 and December 2020. A total of one-hundred and fifty stool samples were collected. All stool samples were examined using the modified Ziehl Neelsen (mZN) staining technique and then examined microscopically for Cryptosporidium infection. RESULTS: A total of 150 children were examined out of which 70 presented with diarrhea. A greater prevalence of 19/70 (27.1%) of Cryptosporidium was observed in children with diarrhea than children without diarrhea 7/80 (8.8%). There was a significant relationship between the prevalence of Cryptosporidium and the presence of diarrhea in children under 5 years in the Kosti Teaching Hospital(P < 0.05). It was found that a higher prevalence was registered among children using piped-water sources for drinking. CONCLUSIONS: The overall prevalence of parasite detected was 17.3% among children admitted to Kosti Teaching Hospital. The prevalence rate of the infection among Children with diarrhoea was 27.1%. Studying the prevalence rate of cryptosporidiosis among diarrheic children may predict their health status, leading to a better diagnosis, treatment, and, therefore, patients' status improvement.


Assuntos
Criptosporidiose/diagnóstico , Diarreia/diagnóstico , Animais , Pré-Escolar , Estudos Transversais , Criptosporidiose/complicações , Criptosporidiose/epidemiologia , Cryptosporidium/crescimento & desenvolvimento , Cryptosporidium/isolamento & purificação , Diarreia/complicações , Fezes/parasitologia , Feminino , Hospitalização , Hospitais de Ensino , Humanos , Lactente , Masculino , Oocistos/fisiologia , Prevalência , Sudão/epidemiologia
4.
Parasitol Res ; 120(5): 1837-1844, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33649965

RESUMO

Cryptosporidium is an important intestinal protozoan parasite that causes diarrhoea in humans and animals. To rapidly and specifically detect Cryptosporidium spp., we designed a pair of primers based on the small subunit ribosomal RNA (SSU rRNA) gene of Cryptosporidium spp. to be used in a new nanoparticle-assisted PCR (nano-PCR) assay. The minimum detectable concentration (1.02 pg) of this nano-PCR was 10 times more sensitive than conventional PCR using the same primer pair. The DNA samples of C. parvum, C. baileyi, C. xiaoi, C. ryanae, and C. andersoni were successfully detected by the nano-PCR. No amplifications were evident with DNA samples of some common intestinal pathogens, including Eimeria tenella, Blastocystis sp., Giardia lamblia, Enterocytozoon bieneusi, and Balantidium coli. To validate the clinical usefulness of the novel nano-PCR, a total of 40 faecal samples from goats, camels, calves, and chickens were examined. The positive rate of Cryptosporidium spp. was 27.5% (11/40), which was consistent with that of an established nested PCR. These results indicate that the novel nano-PCR assay enables the rapid, specific, and accurate detection of Cryptosporidium infection in animals. The findings provide a technical basis for the clinical diagnosis, prevention, and control of cryptosporidiosis.


Assuntos
Criptosporidiose/diagnóstico , Cryptosporidium/isolamento & purificação , Nanopartículas , Reação em Cadeia da Polimerase/métodos , Animais , Camelus , Bovinos , Galinhas , Criptosporidiose/parasitologia , Cryptosporidium/genética , Cryptosporidium parvum/genética , DNA de Protozoário , Fezes/parasitologia , Cabras , Análise de Sequência de DNA
5.
J Parasitol ; 107(2): 163-171, 2021 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-33662118

RESUMO

Cryptosporidium species are the causative agent of cryptosporidiosis and common intracellular parasites that can infect a wide range of vertebrates, including snakes. In previous studies, Cryptosporidium species infections have been reported in snakes in Asia, Europe, and North America. However, limited information is available about the prevalence and molecular characterization of Cryptosporidium in captive snakes in China. Fecal specimens from 609 captive snakes were collected from Beijing (n = 227), Chengdu (n = 12), Dazhou (n = 359), and Ziyang (n = 11). The partial small-subunit (SSU) rRNA gene was amplified by nested polymerase chain reaction to determine the prevalence of Cryptosporidium, and a phylogenetic tree was constructed to assess evolutionary relationships and genetic characteristics. The overall prevalence of Cryptosporidium was 1.97% (12/609). BLAST and phylogenetic analysis of the small subunit ribosomal RNA (SSU rRNA) gene showed that the parasites belonged to Cryptosporidium serpentis. To the best of our knowledge, this is the first study to report the prevalence of Cryptosporidium in snakes of southwestern and northern China and provides preliminary data for the control and prevention of cryptosporidiosis in the investigated areas.


Assuntos
Criptosporidiose/parasitologia , Cryptosporidium/genética , Serpentes/parasitologia , Animais , Animais Domésticos , China/epidemiologia , Criptosporidiose/epidemiologia , Cryptosporidium/classificação , Cryptosporidium/isolamento & purificação , DNA de Protozoário/isolamento & purificação , Fezes/parasitologia , Genótipo , Animais de Estimação , Filogenia , Reação em Cadeia da Polimerase/veterinária , Prevalência , RNA Ribossômico/química , RNA Ribossômico/genética , Análise de Sequência de DNA/veterinária
6.
Parasitol Res ; 120(4): 1281-1289, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33615408

RESUMO

Cryptosporidium is a genus of single-celled protozoa, infecting a wide range of animals and humans. Although Cryptosporidium infections of cattle have been reported in some provinces in China, there is no available information on the prevalence and predominant species of Cryptosporidium in cattle in Jiangxi province. To investigate the prevalence of Cryptosporidium in cattle in Jiangxi province of China, 556 fecal samples were collected from eight farms in four cities and the SSU rRNA locus of Cryptosporidium was amplified from the DNA of each fecal sample by PCR. The overall prevalence of Cryptosporidium was 12.8% (71/556) in cattle in Jiangxi province, with 24.3% (54/222) in Nanchang city, 7.8% (13/166) in Gao'an city, 3.7% (4/108) in Xinyu city, and 0.0% (0/60) in Ji'an city. The differences of the prevalence rates by region, breed, and age groups were statistically significant. All positive PCR products of Cryptosporidium were successfully sequenced and identified as three Cryptosporidium species, namely Cryptosporidium bovis (1/556, 0.18%), Cryptosporidium ryanae (7/556, 1.3%), and Cryptosporidium andersoni (63/556, 11.3%). Furthermore, 36 C. andersoni isolates were successfully classified into three MLST (multilocus sequence typing) subtypes based on four genetic loci (MS1, MS2, MS3, and MS16). The predominant MLST subtype was A4, A4, A4, A1 (n = 30). These findings not only revealed the prevalence and predominant species of Cryptosporidium in cattle in Jiangxi province, but also provided a baseline for studying the genetic structure of C. andersoni, offering a novel resource for better understanding of the epidemiology of Cryptosporidium infection in cattle in Jiangxi province, southeastern China.


Assuntos
Doenças dos Bovinos/epidemiologia , Criptosporidiose/epidemiologia , Cryptosporidium/genética , Cryptosporidium/isolamento & purificação , Animais , Sequência de Bases , Bovinos , Doenças dos Bovinos/parasitologia , China/epidemiologia , Criptosporidiose/parasitologia , Cryptosporidium/classificação , DNA de Protozoário/genética , Fezes/parasitologia , Genótipo , Tipagem de Sequências Multilocus , Filogenia , Reação em Cadeia da Polimerase , Prevalência
7.
Braz J Microbiol ; 52(2): 977-988, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33575990

RESUMO

Neonatal calf diarrhea (NCD) and mortality cause significant losses to the dairy industry. The preweaning dairy calf mortality risk in Uruguay is high (15.2%); however, causes for these losses are largely unknown. This study aimed to assess whether various pathogens were associated with NCD and death in Uruguayan dairy calves and whether these infections, diarrhea, or deaths were associated with the failure of transfer of passive immunity (FTPI). Contemporary diarrheic (n = 264,) and non-diarrheic (n = 271) 1- to 30-day-old calves from 27 farms were sampled. Feces were analyzed by antigen-capture ELISA for Cryptosporidium spp., rotavirus, bovine coronavirus, and Escherichia coli F5+, RT-PCR for bovine astrovirus (BoAstV), and bacterial cultures for Salmonella enterica. Blood/serum was analyzed by RT-PCR or antigen-capture ELISA for bovine viral diarrhea virus (BVDV). Serum of ≤ 8-day-old calves (n = 95) was assessed by refractometry to determine the concention of serum total proteins (STP) as an indicator of FTPI. Whether the sampled calves died before weaning was recorded. At least one pathogen was detected in 65.4% of the calves, and this percentage was significantly higher in diarrheic (83.7%) versus non-diarrheic (47.6%) calves. Unlike the other pathogens, Cryptosporidium spp. and rotavirus were associated with NCD. Diarrheic calves, calves infected with any of the pathogens, and calves infected with rotavirus had significantly lower concentrations of STP. Diarrheic calves had higher chances of dying before weaning than non-diarrheic calves. Diarrheic calves infected with S. enterica were at increased risk of mortality. Controlling NCD, salmonellosis, cryptosporidiosis, and rotavirus infections, and improving colostrum management practices would help to reduce calf morbi-mortality in dairy farms in Uruguay.


Assuntos
Doenças dos Bovinos/etiologia , Doenças dos Bovinos/mortalidade , Diarreia/veterinária , Animais , Animais Recém-Nascidos , Astroviridae/isolamento & purificação , Estudos de Casos e Controles , Bovinos , Cryptosporidium/isolamento & purificação , Indústria de Laticínios/métodos , Diarreia/etiologia , Diarreia/mortalidade , Vírus da Diarreia Viral Bovina/isolamento & purificação , Ensaio de Imunoadsorção Enzimática/veterinária , Escherichia coli/isolamento & purificação , Fezes/microbiologia , Fezes/parasitologia , Fezes/virologia , Feminino , Imunização Passiva/veterinária , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Rotavirus/isolamento & purificação , Salmonella enterica/isolamento & purificação , Inquéritos e Questionários , Síndrome , Uruguai/epidemiologia
8.
BMC Infect Dis ; 21(1): 9, 2021 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-33407180

RESUMO

BACKGROUND: To determine the prevalence of enteric infections in Aboriginal children aged 0-2 years using conventional and molecular diagnostic techniques and to explore associations between the presence of pathogens and child growth. METHODS: Cross-sectional analysis of Aboriginal children (n = 62) residing in a remote community in Northern Australia, conducted from July 24th - October 30th 2017. Stool samples were analysed for organisms by microscopy (directly in the field and following fixation and storage in sodium-acetate formalin), and by qualitative PCR for viruses, bacteria and parasites and serology for Strongyloides-specific IgG. Child growth (height and weight) was measured and z scores calculated according to WHO growth standards. RESULTS: Nearly 60% of children had evidence for at least one enteric pathogen in their stool (37/62). The highest burden of infection was with adenovirus/sapovirus (22.9%), followed by astrovirus (9.8%) and Cryptosporidium hominis/parvum (8.2%). Non-pathogenic organisms were detected in 22.5% of children. Ten percent of children had diarrhea at the time of stool collection. Infection with two or more pathogens was negatively associated with height for age z scores (- 1.34, 95% CI - 2.61 to - 0.07), as was carriage of the non-pathogen Blastocystis hominis (- 2.05, 95% CI - 3.55 to - 0.54). CONCLUSIONS: Infants and toddlers living in this remote Northern Australian Aboriginal community had a high burden of enteric pathogens and non-pathogens. The association between carriage of pathogens/non-pathogens with impaired child growth in the critical first 1000 days of life has implications for healthy child growth and development and warrants further investigation. These findings have relevance for many other First Nations Communities that face many of the same challenges with regard to poverty, infections, and malnutrition.


Assuntos
Infecções por Adenovirus Humanos/epidemiologia , Adenovírus Humanos/genética , Infecções por Astroviridae/epidemiologia , Infecções por Caliciviridae/epidemiologia , Criptosporidiose/epidemiologia , Cryptosporidium/genética , Gastroenterite/epidemiologia , Mamastrovirus/genética , Sapovirus/genética , Infecções por Adenovirus Humanos/virologia , Adenovírus Humanos/isolamento & purificação , Animais , Infecções por Astroviridae/virologia , Austrália/epidemiologia , Infecções por Caliciviridae/virologia , Pré-Escolar , Estudos Transversais , Criptosporidiose/parasitologia , Cryptosporidium/isolamento & purificação , Diarreia/epidemiologia , Diarreia/parasitologia , Diarreia/virologia , Fezes/parasitologia , Fezes/virologia , Feminino , Gastroenterite/parasitologia , Gastroenterite/virologia , Humanos , Lactente , Recém-Nascido , Masculino , Mamastrovirus/isolamento & purificação , Grupo com Ancestrais Oceânicos , Reação em Cadeia da Polimerase/métodos , Prevalência , Sapovirus/isolamento & purificação
9.
BMC Infect Dis ; 21(1): 7, 2021 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-33407198

RESUMO

BACKGROUND: Little is known about the etiology of childhood diarrhea in the United Arab Emirates (UAE) especially after the introduction of rotavirus vaccines. This study aimed to identify gastrointestinal pathogens in children with diarrhea (cases) and the carriage rate of these pathogens in asymptomatic children (controls). METHODS: Stool samples were collected from 203 cases and 73 controls who presented to two major hospitals in Al Ain city, UAE. Samples were analyzed with Allplex™ Gastrointestinal Full Panel Assay for common entero-pathogens. The association between diarrhea and the isolated pathogens was calculated in a multivariate logistic regression model. The adjusted attributable fractions (aAFs) were calculated for all pathogens significantly associated with cases. RESULTS: At least one pathogen was identified in 87 samples (42.8%) from cases and 17 (23.3%) from controls (P < 0.001). Rotavirus, norovirus GII and adenovirus were significantly more prevalent in cases. Their aAFs with 95% ci are 0.95 (0.64, 1.00) for rotavirus, 0.86 (0.38, 0.97) for norovirus GII and 0.84 (0.29, 0.96) for adenovirus. None of the 13 bacteria tested for were more commonly found in the cases than in controls. Cryptosporidium spp. were more significantly detected in cases than in controls. Co-infections occurred in 27.9% of the children. Viruses and parasites were significantly more likely to occur together only in the cases. CONCLUSIONS: Multiplex PCR revealed high positivity rates in both cases and controls which demand a cautious interpretation. Rotavirus remains the main childhood diarrhea pathogen in UAE. Effective strategies are needed to better control rotavirus and other causative pathogens.


Assuntos
Infecções por Adenovirus Humanos/epidemiologia , Adenovírus Humanos/genética , Infecções por Caliciviridae/epidemiologia , Coinfecção/epidemiologia , Criptosporidiose/epidemiologia , Cryptosporidium/genética , Diarreia/epidemiologia , Norovirus/genética , Infecções por Rotavirus/epidemiologia , Rotavirus/genética , Infecções por Adenovirus Humanos/virologia , Adenovírus Humanos/isolamento & purificação , Animais , Infecções por Caliciviridae/virologia , Estudos de Casos e Controles , Pré-Escolar , Coinfecção/parasitologia , Coinfecção/virologia , Criptosporidiose/parasitologia , Cryptosporidium/isolamento & purificação , Diarreia/parasitologia , Diarreia/virologia , Fezes/parasitologia , Fezes/virologia , Feminino , Humanos , Lactente , Masculino , Reação em Cadeia da Polimerase Multiplex/métodos , Norovirus/isolamento & purificação , Rotavirus/isolamento & purificação , Infecções por Rotavirus/virologia , Vacinas contra Rotavirus , Emirados Árabes Unidos/epidemiologia
10.
Parasitol Res ; 120(2): 615-628, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33415392

RESUMO

Cryptosporidium and Giardia are important intestinal zoonotic pathogens that can infect various hosts and cause diarrhoeal diseases. There are few reports of the epidemiological prevalence and molecular characterization of Cryptosporidium and Giardia in wild birds around Qinghai Lake and in the surrounding areas on the Qinghai-Tibetan Plateau, Northwest China. Therefore, the aim of this study was to determine the Cryptosporidium spp. and Giardia duodenalis genotypes and their epidemiological prevalence in wild birds by PCR amplification. To our knowledge, this is the first report of a variety of Cryptosporidium spp. and G. duodenalis infections in wild birds from that area, with overall prevalence rates of 8.98% (61/679) and 3.39% (23/679), respectively. Furthermore, PCR sequencing confirmed the presence of Cryptosporidium baileyi (n = 3), Cryptosporidium parvum (n = 58), and G. duodenalis assemblage B (n = 19) and E (n = 4) in wild birds from the areas around Qinghai Lake. The results of the present study demonstrated the wide distribution of Cryptosporidium and Giardia among wild birds, which has potential public health significance. Moreover, the study findings also provided useful molecular epidemiological data for monitoring and investigating the two parasitic protozoa in wild animals and surrounding environments.


Assuntos
Doenças das Aves/epidemiologia , Criptosporidiose/parasitologia , Cryptosporidium/isolamento & purificação , Diarreia/veterinária , Giardia lamblia/isolamento & purificação , Giardíase/veterinária , Animais , Animais Selvagens , Doenças das Aves/parasitologia , Aves , Criptosporidiose/epidemiologia , Cryptosporidium/genética , Diarreia/epidemiologia , Diarreia/parasitologia , Genótipo , Giardia lamblia/genética , Giardíase/epidemiologia , Giardíase/parasitologia , Lagos , Epidemiologia Molecular , Reação em Cadeia da Polimerase/veterinária , Prevalência , Tibet/epidemiologia
11.
Parasitol Res ; 120(2): 629-635, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33415396

RESUMO

This study evaluated the technology of detection of Giardia spp. cysts and Cryptosporidium spp. oocysts in environmental matrices obtained after water treatment on a bench scale. Calcium carbonate flocculation with immunomagnetic separation was the selected method to quantify the protozoa, and the importance of the number of acid dissociations in the immunomagnetic separation was assessed. When adding the third acid dissociation, an increase of 71% ± 6 in floated residue and 31.9% ± 28.7 in filter backwash water in cyst recovery was observed, while in oocyst recovery, a non-significant increase was detected. In the filtered water, this increased dissociation was important in the protozoa recovery with increases greater than 33%. The results showed that there is a strong interaction of these target organisms with the magnetic microspheres, since protozoa were still recovered in the third acid dissociation and some of them were still adhered to the magnetic microspheres.


Assuntos
Cryptosporidium/isolamento & purificação , Giardia/isolamento & purificação , Purificação da Água/métodos , Água/parasitologia , Animais , Carbonato de Cálcio/química , Floculação , Separação Imunomagnética , Oocistos/isolamento & purificação
12.
J Med Microbiol ; 70(3)2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33507141

RESUMO

Cryptosporidium species are responsible for causing the majority of parasite-related gastrointestinal infections in the UK. This report describes an outbreak of 12 laboratory-confirmed cryptosporidiosis cases identified as part of a Scottish swimming pool investigation, with 9 primary and 3 secondary cases occurring over an 8-week period. Molecular speciation was successful for 11/12 cases, which revealed 10 Cryptosporidium hominis cases and 1 Cryptosporidium parvum case. Of the 10 C. hominis cases, further typing identified 7 as being an unusual sub-type, IbA6G3, which is the first description in the UK of this rare variant. The remaining three C. hominis cases were identified as the common IbA10G2 subtype. Following implementation of control measures on two occasions, no further cases were reported. This report highlights the importance of molecular typing to identify and characterize outbreaks, and emphasizes the need to adhere to swimming pool guidance. It also raises awareness of the potential for outbreaks to involve multiple species/sub-types, and emphasizes the importance of strong public health leadership to ensure effective multi-agency investigations and management of outbreaks.


Assuntos
Criptosporidiose , Cryptosporidium/isolamento & purificação , Surtos de Doenças , Piscinas , Criptosporidiose/epidemiologia , Criptosporidiose/parasitologia , Cryptosporidium/classificação , Humanos , Tipagem Molecular , Escócia
13.
Appl Environ Microbiol ; 86(23)2020 11 10.
Artigo em Inglês | MEDLINE | ID: mdl-32978132

RESUMO

This study investigated the use of biofilms to monitor Cryptosporidium in water. Benthic rock and submersible slide biofilms were sampled upstream and downstream of point sources in a suburban watershed in southeastern Pennsylvania. More oocysts were detected in biofilms scraped from rocks downstream than upstream of a wastewater treatment plant (WWTP) (19 versus 5, respectively; n = 1). Although not statistically significant, Cryptosporidium oocysts were detected more frequently, and in greater numbers, in biofilms grown on slides downstream than upstream of this same WWTP (83.3% positive samples [n = 12] versus 45.5% positive samples [n = 11], respectively; P = 0.0567). Similarly, Cryptosporidium oocysts were detected more frequently, and in greater numbers, in rock biofilms collected downstream than upstream of a stormwater outfall impacted by defective sewer laterals (50% positive samples downstream and 17% positive samples upstream; n = 6; P = 0.2207). While oocyst detection data obtained by slide biofilms versus filters did not necessarily agree on a given day, there was no seasonal difference in the frequency of oocyst detection (P > 0.05) or numbers of oocysts detected (P > 0.05) whether the water was monitored by filtration or slide biofilm sampling. Within any given season, there was no difference in the frequency of oocyst detection (P > 0.05) or the numbers of oocysts detected (P > 0.05) whether the water was monitored by filtration or slide biofilm sampling. These data show that oocyst detection in biofilms is comparable to oocyst detection in filtered water samples. Biofilm sampling offers significant cost savings compared to the filtration-based EPA Method 1623.1 and could be used to identify watershed locations at potential risk for increased oocyst loads.IMPORTANCE Monitoring Cryptosporidium occurrence in watersheds that provide drinking water is necessary to determine where limited resources should most effectively be directed to protect consumers from waterborne exposure to pathogenic oocysts. Biofilms are a useful tool to monitor complex watersheds and identify point sources of Cryptosporidium oocyst contamination that need to be managed to protect public health. Compared to EPA Method 1623.1, the cost benefit of using biofilms to monitor for Cryptosporidium contamination will enable utilities to sample water supplies more frequently, and at more locations, than is currently possible given limited operating budgets. Biofilm sampling could be used to identify high-risk regions within a large, complex watershed and the associated water treatment plants at potential risk for increased oocyst loads in the water supply; this information could then be used to select the locations within the watershed where the more expensive EPA Method 1623.1 is warranted.


Assuntos
Biofilmes , Cryptosporidium/isolamento & purificação , Água Doce/parasitologia , Cryptosporidium/fisiologia , Água Potável/parasitologia , Oocistos/isolamento & purificação , Pennsylvania , Abastecimento de Água
14.
Parasitol Res ; 119(11): 3659-3673, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32960371

RESUMO

Alongside exotic reptiles, amphibians, such as toads, frogs, salamanders, and newts, are nowadays considered popular pets worldwide. As reported for other exotic pet animals, amphibians are known to harbor numerous gastrointestinal parasites. Nonetheless, very little data are available on captive amphibian parasitic diseases. In this study, we applied direct saline fecal smears (DSFS) to examine in total 161 stool samples from 41 different amphibian species belonging to the orders Anura and Caudata. In addition, carbolfuchsin-smear (CFS) staining (n = 74 samples) was used to detect amphibian Cryptosporidium oocysts. Also, complete dissections of deceased amphibians (n = 107) were performed to specify parasite infections and to address parasite-associated pathogenicity. Overall, examined amphibian fecal samples contained 12 different parasite taxa. The order Rhabditida with the species Rhabdias spp. and Strongyloides spp. were the most prevalent nematode species (19.3%), followed by flagellated protozoans (8.7%), Amphibiocapillaria spp./Neocapillaria spp. (7.5%), Oswaldocruzia spp. (4.3%), Blastocystis spp. (3.1%), Cosmocerca spp. (3.1%), oxyurids (Pharyngonoidae) (3.1%), spirurids (1.2%), un-sporulated coccidian oocysts (0.6%), Tritrichomonas spp. (0.6%), Karotomorpha spp. (0.6%), and Cryptosporidium spp. (0.6%). One CFS-stained fecal sample (1.4%) was positive for Cryptosporidium oocysts. Within dissected amphibians, 31 (48.4%) of the anurans and 11 (26.2%) of the salamanders were infected with gastrointestinal parasites. One cutaneous Pseudocapillaroides xenopi infection was diagnosed in an adult African clawed frog (Xenopus laevis). Etiologically, 17 (15.9%) of them died due to severe parasitic and/or bacterial infections (e.g., Chryseobacterium indologenes, Citrobacter freudii, Sphingobacterium multivorum, Klebsiella pneumoniae). High prevalence and pathological findings of several clinical amphibian parasitoses call for more detailed investigation on gastrointestinal parasite-derived molecular mechanisms associated with detrimental lesions or even death.


Assuntos
Animais Exóticos , Animais de Zoológico/parasitologia , Anuros/parasitologia , Enteropatias Parasitárias/veterinária , Urodelos/parasitologia , Animais , Anuros/microbiologia , Blastocystis/isolamento & purificação , Chryseobacterium/isolamento & purificação , Criptosporidiose/parasitologia , Cryptosporidium/isolamento & purificação , Fezes/parasitologia , Feminino , Enteropatias Parasitárias/parasitologia , Nematoides/isolamento & purificação , Oocistos , Sphingobacterium , Strongyloides/isolamento & purificação , Urodelos/microbiologia
15.
PLoS Negl Trop Dis ; 14(9): e0008560, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32898180

RESUMO

BACKGROUND: Food and water-borne illness caused by ingestion of (oo)cysts of Cryptosporidium and Giardia is one of the major health problems globally. Several methods are available to detect Giardia cyst and Cryptosporidium oocyst in food and water. Most of the available methods require a good laboratory facility and well-trained manpower and are therefore costly. There is a need of affordable and reliable method that can be easily implemented in resource limited settings. METHODOLOGY/PRINCIPLE FINDINGS: We developed a smartphone based microscopic assay method to screen (oo)cysts of Cryptosporidium and Giardia contamination of vegetable and water samples. The method consisting of a ball lens of 1 mm diameter, white LED as illumination source and Lugols's iodine staining provided magnification and contrast capable of distinguishing (oo)cysts of Cryptosporidium and Giardia. The analytical performance of the method was tested by spike recovery experiments. The spike recovery experiments performed on cabbage, carrot, cucumber, radish, tomatoes, and water resulted in 26.8±10.3, 40.1±8.5, 44.4±7.3, 47.6±11.3, 49.2 ±10.9, and 30.2±7.9% recovery for Cryptosporidium, respectively and 10.2±4.0, 14.1±7.3, 24.2±12.1, 23.2±13.7, 17.1±13.9, and 37.6±2.4% recovery for Giardia, respectively. The spike recovery results are comparable with data obtained using commercial brightfield and fluorescence microscope methods. Finally, we tested the smartphone microscope system for detecting (oo)cysts on 7 types of vegetable (n = 196) and river water (n = 18) samples. Forty-two percent vegetable and thirty-nine percent water samples were found to be contaminated with Cryptosporidium oocyst. Similarly, thirty-one percent vegetable and thirty-three percent water samples were contaminated with Giardia cyst. CONCLUSIONS: The newly developed smartphone microscopic method showed comparable performance to commercial microscopic methods. The new method can be a low-cost and easy to implement alternative method for simultaneous detection of (oo)cysts in vegetable and water samples in resource limited settings.


Assuntos
Cryptosporidium/isolamento & purificação , Doenças Transmitidas por Alimentos/prevenção & controle , Giardia/isolamento & purificação , Oocistos/isolamento & purificação , Imagem Óptica/métodos , Água Potável/parasitologia , Doenças Transmitidas por Alimentos/parasitologia , Humanos , Microscopia de Fluorescência/métodos , Smartphone , Verduras/parasitologia
16.
Folia Parasitol (Praha) ; 672020 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-32812530

RESUMO

Although the microscopic examination of stool samples remains the reference method of choice for the diagnosis of intestinal protistan infections, this method is time-consuming and requires experienced and well-trained operators. The purpose of this study was to evaluate the level of agreement between the BD MAX TM Enteric Parasite Panel (EPP) and microscopy for the detection of Giardia intestinalis (Lambl, 1859), Cryptosporidium spp. and Entamoeba histolytica Schaudinn, 1903 in stool samples. The study included faecal samples of 362 patients who were admitted to our hospital due to gastrointestinal complaints. In the microscopic examination, which was made with the native-lugol method on the stool samples that were taken from the patients, cysts, trophozoites and eggs of the parasite were examined. The diagnosis of G. intestinalis, Cryptosporidium parvum Tyzzer, 1912 and Cryptosporidium hominis Morgan-Ryan, Fall, Ward, Hijjawi, Sulaiman, Fayer, Thompson, Olson, Lal et Xiao, 2002, and E. histolytica was made in the faecal samples using the EPP assay. In the microscopic examination, Cryptosporidium spp. positive stool samples were stained with kinyoun's acid-fast. In the microscopic examination, parasites were detected in 41 (11%) of the 362 stool samples. In contrast, EPP assay identified parasites in 23 (6.3%) of the samples. In the microscopic examination, E. histolytica and Entamoeba dispar Brumpt, 1925 were detected in 22 (6.1%) of the samples, G. intestinalis was seen in 15 (4.1%), and C. parvum or C. hominis were detected in three (0.8%); these values were five (1.4%), 16 (4.4%) and two (0.5%) positive with the EPP assay. Although C. parvum or C. hominis were detected as positive in the microscopic examination of three samples, only two of the samples were positive in both EPP assay and kinyoun's acid-fast method. The EPP assay is a relatively simple test that can distinguish E. histolytica and E. dispar, but it cannot replace microscopy in the diagnosis of amoebiasis. Diagnosis for G. intestinalis and C. parvum/C. hominis with the BD MAXTM enteric parasite panel was equivalent to that with microscopy. We believe that E. histolytica must be diagnosed with nucleic acid amplification tests that have a high sensitivity and specificity like EPP assay in certain patient groups.


Assuntos
Criptosporidiose/epidemiologia , Entamebíase/epidemiologia , Fezes/parasitologia , Giardíase/epidemiologia , Adulto , Criptosporidiose/parasitologia , Cryptosporidium/isolamento & purificação , Cryptosporidium parvum/isolamento & purificação , Entamoeba histolytica/isolamento & purificação , Entamebíase/parasitologia , Feminino , Giardia lamblia/isolamento & purificação , Giardíase/parasitologia , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Turquia/epidemiologia
17.
Parasitol Res ; 119(9): 3119-3123, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32743725

RESUMO

Few reports of Cryptosporidium spp. in snakes in China have been published. To determine the infection rate and document the presence of Cryptosporidium in pet snakes using molecular methods, 273 fecal samples were collected from eight species of pet snakes from 13 pet households in Beijing, China, and were examined by PCR amplification of the small subunit ribosomal RNA gene. Cryptosporidium was detected from 17 of 273 (6.2%) samples, and nine out of 13 households tested positive for Cryptosporidium with a range of 3.3 to 33.3% among households showing significant difference (p < 0.01). The infection rate of Cryptosporidium for females and males was 6.5% (13/201) and 5.6% (4/72), respectively, showing no significant difference (p > 0.05). Six out of eight pet snake species tested positive for Cryptosporidium with a range of 4.2 to 9.1% among species, showing no significant difference (p > 0.05). Two Cryptosporidium species were identified: Cryptosporidium serpentis in 10 samples and Cryptosporidium varanii in seven samples. No zoonotic Cryptosporidium species occur in our study populations.


Assuntos
Criptosporidiose/parasitologia , Cryptosporidium/isolamento & purificação , Animais de Estimação/parasitologia , Serpentes/parasitologia , Animais , Pequim , Cryptosporidium/classificação , Cryptosporidium/genética , Fezes/parasitologia , Reação em Cadeia da Polimerase
18.
Parasitol Res ; 119(9): 3033-3040, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32748039

RESUMO

We assessed the potential contribution of hospitals to contaminations of wastewater by enteric protists, including Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi in raw wastewater. Wastewater samples were collected from storage tanks in two hospitals and one associated wastewater treatment plant in Shanghai, China, from March to November 2009. Enteric pathogens were detected and identified using PCR and DNA sequencing techniques. Among a total of 164 samples analyzed, 31 (18.9%), 45 (27.4%), and 122 (74.4%) were positive for Cryptosporidium spp., G. duodenalis, and E. bieneusi, respectively. Altogether, three Cryptosporidium species, four G. duodenalis assemblages, and 12 E. bieneusi genotypes were detected. Cryptosporidium hominis, G. duodenalis sub-assemblage AII, and E. bieneusi genotype D were the dominant ones in wastewater from both hospitals and the wastewater treatment plant. A similar distribution in genotypes of enteric pathogens was seen between samples from hospitals and the wastewater treatment plant, suggesting that humans are one of the major sources for these pathogens and hospitals are important contributors of enteric parasites in urban wastewater. Data from this study might be useful in the formulation of preventive measures against environmental contamination of waterborne pathogens.


Assuntos
Infecção Hospitalar/microbiologia , Infecção Hospitalar/parasitologia , Cryptosporidium/isolamento & purificação , Enterocytozoon/isolamento & purificação , Giardia lamblia/isolamento & purificação , Águas Residuárias/microbiologia , Águas Residuárias/parasitologia , China/epidemiologia , Infecção Hospitalar/epidemiologia , Criptosporidiose/epidemiologia , Criptosporidiose/parasitologia , Cryptosporidium/classificação , Cryptosporidium/genética , Enterocytozoon/classificação , Enterocytozoon/genética , Genótipo , Giardia lamblia/classificação , Giardia lamblia/genética , Giardíase/epidemiologia , Giardíase/parasitologia , Hospitais , Humanos , Microsporidiose/epidemiologia , Microsporidiose/microbiologia , Reação em Cadeia da Polimerase
19.
Exp Parasitol ; 218: 107967, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32858044

RESUMO

Reported incidence rates of cryptosporidiosis in Ireland are consistently among the highest in Europe. Despite the national prevalence of this enteric parasite and the compulsory nature of incidence surveillance and reporting, in-depth analyses seeking to genotype clinical isolates of Cryptosporidium on an intra-species level are rarely undertaken in Ireland. This molecular epidemiology study of 163 clinical Cryptosporidium isolates was conducted in Southern Ireland, from 2015 to 2018, in order to ascertain population subtype heterogeneity. Analysis was conducted via real-time PCR amplification and gp60 gene sequencing, which successfully determined the subtype designation of 149 of the 163 (91.4%) tested isolates. Overall, 12 C. parvum and five C. hominis subtypes were identified, with the incidence of the regionally predominant C. parvum species found to primarily occur during springtime months, while C. hominis incidence was largely confined to late summer and autumnal months. Additionally, one C. parvum and four C. hominis subtypes were newly reported by this study, having not been previously identified in clinical or livestock infection in Ireland. Overall, these data give insight into the diversification of the Cryptosporidium population and emergent subtypes, while also allowing comparisons to be made with clinical epidemiological profiles reported previously in Ireland and elsewhere.


Assuntos
Criptosporidiose/parasitologia , Cryptosporidium/classificação , Zoonoses/parasitologia , Animais , Criptosporidiose/epidemiologia , Cryptosporidium/genética , Cryptosporidium/isolamento & purificação , Cryptosporidium parvum/classificação , Cryptosporidium parvum/genética , Cryptosporidium parvum/isolamento & purificação , DNA de Protozoário/isolamento & purificação , Fezes/parasitologia , Gastroenterite/parasitologia , Glicoproteínas/química , Glicoproteínas/genética , Humanos , Incidência , Irlanda/epidemiologia , Estudos Longitudinais , Prevalência , Proteínas de Protozoários/química , Proteínas de Protozoários/genética , RNA Ribossômico 18S/genética , Reação em Cadeia da Polimerase em Tempo Real , Estudos Retrospectivos , Estações do Ano , Alinhamento de Sequência
20.
PLoS Negl Trop Dis ; 14(7): e0008448, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32658930

RESUMO

BACKGROUND: Cryptosporidium is a protozoan parasite that causes mild to severe diarrhoeal disease in humans. To date, several commercial companies have developed rapid immunoassays for the detection of Cryptosporidium infection. However, the challenge is to identify an accurate, simple and rapid diagnostic tool for the estimation of cryptosporidiosis burden. This study aims at evaluating the accuracy of CerTest Crypto, a commercialized rapid diagnostic test (RDT) for the detection of Cryptosporidium antigens in the stool of children presenting with diarrhoea. METHODS: A cross-sectional study was conducted in four study sites in Sub-Saharan Africa (Gabon, Ghana, Madagascar, and Tanzania), from May 2017 to April 2018. Stool samples were collected from children under 5 years with diarrhoea or a history of diarrhoea within the last 24 hours. All specimens were processed and analyzed using CerTest Crypto RDT against a composite diagnostic panel involving two polymerase chain reaction (PCR) tests (qPCR and RFLP-PCR,) as the gold standard. RESULTS: A total of 596 stool samples were collected. Evaluation of the RDT yielded a very low overall sensitivity of 49.6% (confidence interval (CI) 40.1-59.0), a specificity of 92.5% (CI 89.8-94.7), positive predictive value of 61.3% (CI 50.6-71.2), and negative predictive value of 88.5% (85.3-91.1) when compared to the composite reference standard of qPCR and RFLP-PCR for the detection of Cryptosporidium species. Moreover, the performance of this test varied across different sites. CONCLUSION: The weak performance of the studied RDT suggests the need to carefully evaluate available commercial RDTs before their use as standard tools in clinical trials and community survey of Cryptosporidium infections in pediatric cohorts.


Assuntos
Criptosporidiose/diagnóstico , Cryptosporidium/isolamento & purificação , Diarreia/parasitologia , África ao Sul do Saara/epidemiologia , Pré-Escolar , Estudos Transversais , Criptosporidiose/epidemiologia , Fezes/parasitologia , Feminino , Humanos , Lactente , Masculino , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Fragmento de Restrição , Sensibilidade e Especificidade
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