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1.
Acta Trop ; 233: 106568, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35716763

RESUMO

Microsporidia are obligate intracellular fungus-like parasites that infect humans and animals worldwide. However, there is limited epidemiological data on the occurrence and molecular diversity of microsporidia in buffaloes worldwide. In the present study, fecal samples of 300 water buffaloes (Bubalus bubalis) in Kayseri, Sivas, and Samsun provinces of Turkey were investigated using two nested PCR assays targeting the rRNA of E. bieneusi and Encephalitozoon spp. All the fecal samples from water buffalo were found to be negative for Encephalitozoon spp. PCR positive isolates of E. bieneusi were bidirectionally sequenced for genotyping and phylogenetic analyses. Enterocytozoon bieneusi was the only microsporidian species identified in 8 water buffaloes with an overall molecular prevalence of 2.7%. Two known genotypes, YNDCEB-90 (n = 5) and J (n = 3) were identified by ITS sequence analysis. The YNDCEB-90 and J genotypes fall into zoonotic Group 1 and 2 of E. bieneusi in the phylogenetic tree, respectively. These findings suggested that water buffalo in Turkey are harbouring zoonotic genotypes of E. bieneusi and may have a significant risk for zoonotic transmission to humans. This is the first report of detecting E. bieneusi genotypes J and YNDCEB-90 in water buffaloes. Further insight into the epidemiology of E. bieneusi in water buffaloes in different geographical areas in Turkey will be highly important to have determined the public health significance of this pathogen.


Assuntos
Encephalitozoon , Enterocytozoon , Microsporídios , Microsporidiose , Animais , Búfalos , China/epidemiologia , Enterocytozoon/genética , Fezes/parasitologia , Genótipo , Humanos , Microsporidiose/epidemiologia , Microsporidiose/veterinária , Filogenia , Prevalência , Turquia/epidemiologia
2.
Ann Parasitol ; 68(1): 87-92, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35491745

RESUMO

Microsporidia belong to the intracellular spore-like pathogen, that can cause infection in invertebrates and vertebrates, including humans. Encephalitozoon spp. and Enterocytozoon bieneusi, are important causes of chronic diarrhea, especially in patients with HIV/AIDS. Therefore, in this study, modified trichrome staining (MTS) and nested polymerase chain reaction (nested PCR) methods were used for the diagnosis of common intestinal microsporidia in faecal samples of patients with HIV/AIDS in Zahedan, southeastern Iran, for the first time. Stool samples were collected from 50 HIV/AIDS-infected patients with gastrointestinal symptoms whose infections were confirmed by serology test. Prepared smears from each stool sample were stained using the MTS method. Nested PCR was used to amplify 440 bp and 629 bp fragments of 16S rRNA genes in E. bieneusi and Encephalitozoon spp., respectively. Based on the MTS method and the nested PCR, 8 (16%) and 12 (24%) stool samples were positive, respectively. According to the results of nested PCR, eight, three, and one case were infected with E. bieneusi, Encephalitozoon spp., and both of them, respectively. Findings indicated microsporidiosis in HIV/AIDS-infected patients in Zahedan is an important health problem. Therefore, this opportunistic microorganism in HIV/AIDS-infected patients should be diagnosed using sensitive and accurate methods.


Assuntos
Encephalitozoon , Infecções por HIV , Microsporídios , Microsporidiose , Animais , Encephalitozoon/genética , Humanos , Irã (Geográfico)/epidemiologia , Microsporídios/genética , Microsporidiose/diagnóstico , Microsporidiose/epidemiologia , RNA Ribossômico 16S
3.
Front Cell Infect Microbiol ; 11: 694957, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34095003

RESUMO

Microsporidia are a group of spore-forming, fungus-related pathogens that can infect both invertebrates and vertebrates including humans. The primary infection site is usually digestive tract, but systemic infections occur as well and cause damages to organs such as lung, brain, and liver. The systemic spread of microsporidia may be intravascular, requiring attachment and colonization in the presence of shear stress. Von Willebrand Factor (VWF) is a large multimeric intravascular protein and the key attachment sites for platelets and coagulation factors. Here in this study, we investigated the interactions between VWF and microsporidia Encephalitozoon hellem (E. hellem), and the modulating effects on E. hellem after VWF binding. Microfluidic assays showed that E. hellem binds to ultra-large VWF strings under shear stress. In vitro germination assay and infection assay proved that E. hellem significantly increased the rates of germination and infection, and these effects would be reversed by VWF blocking antibody. Mass spectrometry analysis further revealed that VWF-incubation altered various aspects of E. hellem including metabolic activity, levels of structural molecules, and protein maturation. Our findings demonstrated that VWF can bind microsporidia in circulation, and modulate its pathogenicity, including promoting germination and infection rate. VWF facilitates microsporidia intravascular spreading and systemic infection.


Assuntos
Encephalitozoon , Microsporídios , Animais , Humanos , Fator de von Willebrand
4.
Parasite Immunol ; 43(6): e12828, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33682117

RESUMO

Microsporidia are a group of obligate, intracellular, spore-forming eukaryotic pathogens, which predominantly infects immunocompromised individuals worldwide. Encephalitozoon spp. is one of the most prevalent microsporidia known to infect humans. Host immune system plays a major role in combating pathogens including Encephalitozoon spp. infecting humans. Both innate and adaptive arms of host immune system work together in combating Encephalitozoon infection. Researchers are conducting studies to elucidate the role of both arms of immune system against Encephalitozoon infection. In addition to cell-mediated adaptive immunity, role of innate immunity is also being highlighted in clearance of Encephalitozoon spp. from host body. Therefore, the current review will give a clear and consolidated update on the role of innate as well as adaptive immunity in protection against Encephalitozoon spp.


Assuntos
Encephalitozoon , Encefalitozoonose , Humanos , Imunidade Celular , Imunidade Inata , Hospedeiro Imunocomprometido
5.
Cytometry A ; 99(4): 343-347, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33704904

RESUMO

Genome size information is sparse across fungi, with information being available for less than 2000 species. So far, most records have been obtained using static, microscope-based cytometry methods or derived from genome sequencing projects. Flow cytometry is now considered the state-of-the-art method for obtaining genome size measurements, and appropriate methods and DNA standards are available, enabling the analysis of most genome size ranges in a rapid, robust and inexpensive way. The average fungal genome size is 60 Mbp, but sizes vary across phylogeny, ranging from 2.2 (Encephalitozoon romaleae) to 3706 Mbp (Jafnea semitosta). In several fungal clades, genome size expansion seems to accompany evolution either to plant mutualism or to plant parasitism (particularly biotrophy), and fungi that interact with plants seem to have larger genomes than saprobes and those that interact with animals. Whereas flow cytometry for nuclear DNA quantification is routinely employed in plant sciences for genome size and ploidy studies, its use in fungal biology is still infrequent. Appropriate standards, methods and best practices are described here, with the aim of stimulating a more generalized and widespread use of flow cytometry for fungal genome size measurement.


Assuntos
Fungos , Genoma de Planta , Animais , Ascomicetos , DNA Fúngico/genética , DNA de Plantas , Encephalitozoon , Citometria de Fluxo , Fungos/genética , Tamanho do Genoma
6.
J Cell Sci ; 134(5)2021 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-33589497

RESUMO

Microsporidia are a large phylum of obligate intracellular parasites. Approximately a dozen species of microsporidia infect humans, where they are responsible for a variety of diseases and occasionally death, especially in immunocompromised individuals. To better understand the impact of microsporidia on human cells, we infected human colonic Caco2 cells with Encephalitozoon intestinalis, and showed that these enterocyte cultures can be used to recapitulate the life cycle of the parasite, including the spread of infection with infective spores. Using transmission electron microscopy, we describe this lifecycle and demonstrate nuclear, mitochondrial and microvillar alterations by this pathogen. We also analyzed the transcriptome of infected cells to reveal host cell signaling alterations upon infection. These high-resolution imaging and transcriptional profiling analysis shed light on the impact of the microsporidial infection on its primary human target cell type.This article has an associated First Person interview with the first authors of the paper.


Assuntos
Encephalitozoon , Células CACO-2 , Encephalitozoon/genética , Enterócitos , Humanos , Transdução de Sinais
7.
J Mol Diagn ; 23(4): 417-423, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33387699

RESUMO

Microsporidiosis and cryptosporidiosis are associated with chronic diarrhea in immunocompromised patients. The objectives of this study were to: i) assess a multiplex quantitative PCR assay targeting Cryptosporidium spp and the microsporidian Enterocytozoon bieneusi and Encephalitozoon spp, and ii) provide an update on the epidemiology of these pathogens. A prospective study was conducted from January 2017 to January 2019. Performance of the assay was assessed, and all cryptosporidia and microsporidia isolates were genotyped. The sensitivity of the multiplex PCR method reached 1 copy/µL for each targeted pathogen. The sensitivity of co-proantigen testing in the diagnosis of cryptosporidiosis was 73%. The sensitivity of microscopy in the diagnosis of cryptosporidiosis was 64%, and microsporidiosis, 50%. Among the 456 patients included, 14 were positive for Cryptosporidium spp (4 different species); 5, for E. bieneusi; and 2, for Encephalitozoon intestinalis. The overall prevalence of cryptosporidia was 3.1%, and of microsporidia, 1.5%; in kidney transplant recipients (n = 82), corresponding values were 7.3% and 2.4% (6 and 2 patients), respectively. Two cases of E. intestinalis infection were diagnosed in children who had traveled to the tropics. This study is the first to assess a multiplex quantitative PCR method for the simultaneous diagnosis of intestinal microsporidiosis and cryptosporidiosis. The highest prevalences of both pathogens were observed in kidney transplant recipients.


Assuntos
Criptosporidiose/diagnóstico , Criptosporidiose/epidemiologia , Cryptosporidium/genética , Encephalitozoon/genética , Enterocytozoon/genética , Microsporidiose/diagnóstico , Microsporidiose/epidemiologia , Reação em Cadeia da Polimerase Multiplex/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Criptosporidiose/parasitologia , Cryptosporidium/isolamento & purificação , Diarreia/microbiologia , Diarreia/parasitologia , Encephalitozoon/isolamento & purificação , Enterocytozoon/isolamento & purificação , Fezes/microbiologia , Fezes/parasitologia , Feminino , França/epidemiologia , Genótipo , Humanos , Hospedeiro Imunocomprometido , Lactente , Recém-Nascido , Masculino , Microsporidiose/microbiologia , Pessoa de Meia-Idade , Prevalência , Estudos Prospectivos , Sensibilidade e Especificidade , Adulto Jovem
8.
Cornea ; 40(2): 242-244, 2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-32826651

RESUMO

PURPOSE: To report a case of microsporidia (Encephalitozoon hellem) keratoconjunctivitis acquired through avian transmission in an immunocompetent adult, diagnosed by metagenomic deep sequencing (MDS), and confirmed by polymerase chain reaction. METHODS: A case report. RESULTS: An 18-year-old woman was referred with unilateral keratoconjunctivitis unresponsive to topical and systemic therapy after exposure to birdcage debris. Slit-lamp examination of the left eye revealed a follicular papillary reaction of the palpebral conjunctiva and multiple corneal punctate epithelial opacities that stained minimally with fluorescein. In vivo confocal microscopy revealed bright double-walled structures and smaller bright round structures in the superficial epithelial debris and epithelium. Molecular diagnosis with MDS of E. hellem was confirmed by polymerase chain reaction. Clinical resolution and normalization of in vivo confocal microscopy was observed after a 6-week course of topical azithromycin. The patient elected a 3-week course of topical voriconazole 1% for definitive antimicrosporidial treatment, with no evidence of persistent infection 1 month later. CONCLUSIONS: Microsporidial (E. hellem) keratoconjunctivitis can occur through avian transmission in immunocompetent hosts. Topical azithromycin may be effective against this pathogen. MDS has utility in the diagnosis of atypical keratoconjunctivitis.


Assuntos
Encephalitozoon/isolamento & purificação , Infecções Oculares Fúngicas/diagnóstico , Ceratoconjuntivite/diagnóstico , Microsporidiose/diagnóstico , Adolescente , Antibacterianos/uso terapêutico , Antifúngicos/uso terapêutico , Azitromicina/uso terapêutico , Quimioterapia Combinada , Encephalitozoon/genética , Infecções Oculares Fúngicas/tratamento farmacológico , Infecções Oculares Fúngicas/microbiologia , Feminino , Humanos , Imunocompetência , Ceratoconjuntivite/tratamento farmacológico , Ceratoconjuntivite/microbiologia , Metagenômica , Microscopia Confocal , Microsporidiose/tratamento farmacológico , Microsporidiose/microbiologia , Reação em Cadeia da Polimerase , Voriconazol/uso terapêutico
9.
Acta Parasitol ; 66(2): 397-405, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33033999

RESUMO

PURPOSE: Encephalitozoon intestinalis affects many physiological processes of host cells to survive, proliferate, and spread to different regions within the body. In this study, the effects of the parasite on host cell apoptosis and proliferation were investigated. METHODS: To determine the impact of the parasite on the host cell apoptosis, changes in the expression profile of genes were investigated with the qPCR array using the Human Apoptosis Panel in infected and non-infected macrophage cells. Also, the rate of apoptosis in the cells was determined by Giemsa staining method. Cell proliferation was determined by measuring the DNA concentration in infected and non-infected cells. RESULTS: The thirty-six of apoptosis-related genes were down-regulated, while 20 of apoptosis-related genes were up-regulated in infected cells compared to uninfected cells. However, there were no significant changes detected in 32 analyzed genes between infected and control groups. E. intestinalis was determined to decrease cell proliferation in U937 macrophage cells. Unexpectedly, Giemsa staining showed an increase in the rate of apoptosis in infected cells. CONCLUSION: Regulated genes after infection are involved in many different biological pathways and various components of the cell. This suggests that the parasite uses highly sophisticated ways to maintain the viability of the cell.


Assuntos
Encephalitozoon , Encefalitozoonose , Apoptose , Humanos , Células U937
10.
Med Mycol J ; 62(3): 59-62, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34471036

RESUMO

To evaluate the role of companion birds as a reservoir of Encephalitozoon hellem infection in humans, the present study determined the prevalence and genotypes of E. hellem from 269 birds in 4 pet shops using polymerase chain reaction (PCR) assay. E. hellem was identified in 4.8% (13/269) of the birds and was detected in all pet shops. Every positive sample corresponded to zoonotic genotype 1A. Considering the low prevalence of E. hellem infection, it is likely that the risk of zoonotic transmission from companion birds kept in pet shops to humans is low in Japan.


Assuntos
Encephalitozoon , Animais , Aves , Fezes , Humanos , Japão
11.
Sci Rep ; 11(1): 17467, 2021 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-34471179

RESUMO

Intestinal parasitic infections have high prevalence rate in many regions especially in developing countries. The aim of this study was to determine the presence and genotype/subtype of some intestinal protozoa in livestock in Iran. Stool samples were collected from cattle, sheep, chickens, and horses. The presence of targeted parasites was evaluated using real-time PCR. Genotyping/subtyping of positive samples was characterized using sequencing of the ITS and barcoding region, respectively. Blastocystis sp., 27.7% (48/173) and Enterocytozoon bieneusi 26.0% (45/173) were the most frequent protozoa followed by Encephalitozoon spp., 0.57% (1/173). Cryptosporidium spp. were not detected among samples. Encephalitozoon spp., was detected only in chickens 2.2% (1/45). A statistically correlation was seen between animals and the prevalence of targeted protozoa. E. bieneusi genotypes I (9/38; 23.68%), BEB6 (22/38; 57.89%), D (6/38; 15.79%), and horse1 (1/38; 2.63%) were detected among samples. A statistically significant correlation was seen between the genotypes and animals (P ≤ 0.05). Blastocystis sp., ST1 (1/45; 2.22%), ST5 3/45; 6.66%), ST7 (1/45; 2.22%), ST10 (24/45; 53.33%), and ST14 (16/45; 35.55%) were characterized among samples. There was no significant correlation between certain subtypes and animals (P = 0.173). The presence of zoonotic potential genotypes of E. bieneusi in animals and zoonotic potential subtypes ST1 and ST7 among our samples provide a clue about the transmission dynamic of E. bieneusi and Blastocystis sp. between animals-animals and humans-animals.


Assuntos
Blastocystis/fisiologia , Encephalitozoon/fisiologia , Enterocytozoon/fisiologia , Zoonoses/epidemiologia , Animais , Blastocystis/genética , Infecções por Blastocystis/epidemiologia , Infecções por Blastocystis/parasitologia , Bovinos , Galinhas , Encephalitozoon/genética , Encefalitozoonose/epidemiologia , Encefalitozoonose/microbiologia , Enterocytozoon/genética , Genótipo , Cavalos , Irã (Geográfico)/epidemiologia , Gado , Microsporidiose/epidemiologia , Microsporidiose/microbiologia , Epidemiologia Molecular , Filogenia , Prevalência , Ovinos , Zoonoses/microbiologia , Zoonoses/parasitologia
12.
Acta Parasitol ; 66(2): 346-353, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32996015

RESUMO

BACKGROUND: Microsporidia infection was originally described as an immunocompromised associated pathogen. Limitations to correct microscopic diagnosis of microsporidia include size of the organism presenting a challenge even to a highly competent laboratory expert. OBJECTIVE: The present study aimed to detect microsporidia infection among leukemic children. The performance of modified trichrome stain and PCR in the diagnosis of microsporidia was evaluated with further speciation. METHODS: Stool samples of 100 leukemic children on chemotherapy were examined microscopically for microsporidia. DNA was extracted from all samples. Amplification was performed by conventional and nested PCR. Sequencing of amplified products was performed on unspeciated samples. RESULTS: Microsporidia were detected in 23% of the children by MTS and 29% by PCR. The 29 positive samples were subjected to PCR for speciation. Enterocytozoon bieneusi was found to predominate in 20 cases, Encephalitozoon intestinalis in three cases, two cases had co-infection, and the remaining four samples were not amplified with either E. bieneusi or E. intestinalis specific primers. By DNA sequencing of the unspeciated samples, three samples shared high homology with Encephalitozoon hellem and one sample with Encephalitozoon cuniculi. Referring to PCR as a gold standard, MTS exhibited 72.4% sensitivity and 97.2% specificity with 90% accuracy. Among a number of studied variables, diarrhea and colic were significantly associated with microsporidia infection when diagnosed by either technique. CONCLUSION: The use of sensitive and discriminative molecular tools will contribute to determining the true prevalence of microsporidiosis and possibly their potential transmission source depending on species identification.


Assuntos
Encephalitozoon , Enterocytozoon , Microsporídios , Microsporidiose , Criança , Fezes , Humanos
13.
BMC Vet Res ; 16(1): 212, 2020 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-32571322

RESUMO

BACKGROUND: Microsporidia are common opportunistic parasites in humans and animals, including rabbits. However, only limited epidemiology data concern about the prevalence and molecular characterization of Enterocytozoon bieneusi and Encephalitozoon spp. in rabbits. This study is the first detection and genotyping of Microsporidia in pet rabbits in China. RESULTS: A total of 584 faecal specimens were collected from rabbits in pet shops from four cities in Sichuan province, China. The overall prevalence of microsporidia infection was 24.8% by nested PCR targeting the internal transcribed spacer (ITS) region of E. bieneusi and Encephalitozoon spp. respectively. E. bieneusi was the most common species (n = 90, 15.4%), followed by Encephalitozoon cuniculi (n = 34, 5.8%) and Encephalitozoon intestinalis (n = 16, 2.7%). Mixed infections (E. bieneusi and E. cuniculi) were detected in five another rabbits (0.9%). Statistically significant differences in the prevalence of microsporidia were observed among different cities (χ2 = 38.376, df = 3, P < 0.01) and the rabbits older than 1 year were more likely to harbour microsporidia infections (χ2 = 9.018, df = 2, P < 0.05). Eleven distinct genotypes of E. bieneusi were obtained, including five known (SC02, I, N, J, CHY1) and six novel genotypes (SCR01, SCR02, SCR04 to SCR07). SC02 was the most prevalent genotype in all tested cities (43.3%, 39/90). Phylogenetic analysis showed that these genotypes were clustered into group 1-3 and group 10. Meanwhile, two genotypes (I and II) were identified by sequence analysis of the ITS region of E. cuniculi. CONCLUSION: To the best of our knowledge, this is the first report of microsporidia infection in pet rabbits in China. Genotype SC02 and four novel genotypes were classified into potential zoonotic group 1, suggesting that pet rabbits may cause microsporidiosis in humans through zoonotic transmissions. These findings provide preliminary reference data for monitoring microsporidia infections in pet rabbits and humans.


Assuntos
Encephalitozoon/isolamento & purificação , Enterocytozoon/isolamento & purificação , Microsporidiose/veterinária , Animais , China/epidemiologia , Encephalitozoon/classificação , Encephalitozoon/genética , Enterocytozoon/classificação , Enterocytozoon/genética , Fezes/microbiologia , Genótipo , Microsporidiose/epidemiologia , Reação em Cadeia da Polimerase/veterinária , Prevalência , Coelhos
14.
Parasitol Res ; 119(6): 1903-1913, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32385710

RESUMO

Microsporidia is a group of spore-forming microorganisms with zoonotic potential. This study aimed to compare intestinal microsporidia infections in cat owners and non-pet owners. In total, 210 fecal samples were collected from indoor cats, cat owners, and non-pet owners. DNA extraction was performed and the small subunit ribosomal RNA (SSU rRNA) gene was amplified. To characterize the genotypes, the internal transcribed spacer (ITS) fragment was amplified and sequenced. The phylogenetic trees were drawn to evaluate the relationship among Enterocytozoon bieneusi isolates. Two (2.9%) and one (1.4%) fecal samples from cat owners and one (1.4%) and two (2.9%) fecal samples from non-pet owners were positive for E. bieneusi and Encephalitozoon intestinalis, respectively. E. bieneusi was detected in two cat samples (2.9%). Same infection was not seen between infected cats and their owners. There was no significant difference between the prevalence rate of microsporidia among the cat owners and non-pet owners. Indeed, the genotypes L and type IV were seen in cats, while the genotype D was only detected in human. In this study, E. bieneusi and E. intestinalis were more prevalent among the cat owners and non-pet owners, respectively. Indeed, the higher prevalence of E. bieneusi in cats and their owners might be resulted from the worldwide distribution of this species.


Assuntos
Doenças do Gato/parasitologia , Enteropatias Parasitárias/parasitologia , Microsporídios , Microsporidiose/diagnóstico , Adulto , Animais , Estudos de Casos e Controles , Doenças do Gato/epidemiologia , Gatos , Encephalitozoon/isolamento & purificação , Enterocytozoon/isolamento & purificação , Fezes/parasitologia , Feminino , Genótipo , Humanos , Enteropatias Parasitárias/veterinária , Irã (Geográfico)/epidemiologia , Masculino , Microsporídios/classificação , Microsporídios/genética , Microsporídios/isolamento & purificação , Microsporidiose/epidemiologia , Microsporidiose/veterinária , Pessoa de Meia-Idade , Animais de Estimação/parasitologia , Filogenia , Prevalência , Zoonoses/epidemiologia
15.
Microb Pathog ; 141: 103988, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31972268

RESUMO

Intestinal microsporidiosis is known as an opportunistic infection in immunocompromised patients. The current study aimed to investigate intestinal microsporidia infection in human subjects with/without immunodeficiency. Totally, 600 stool samples were collected from immunocompromised (254) and immunocompetent (346) subjects. DNA extraction was performed and the SSU rRNA and the ITS genes were amplified to detect and characterize microsporidia and the relevant genotypes. Phylogenetic trees were drawn using MEGA7 software to illustrate the correlation between isolates. From 600 enrolled subjects, 283 and 317 were male and female, respectively. The average age ± SD of all tested subjects was 28.85 ± 26.92. The results of PCR demonstrated the presence of E. bieneusi and Encephalitozoon sp., among 10/600 (1.67%) and 26/600 (4.33%) of samples, respectively. Accordingly, E. bieneusi was seen among 4/346 (1.15%), 1/53 (1.88%), 3/124 (2.42%), and 2/63 (3.17%), and Encephalitozoon sp., was detected from 17/346 (4.91%), 3/53 (5.36%), 4/124 (3.22%) and 2/63 (3.17%) of healthy subjects, RA patients, cancer patients, and transplantation recipients, respectively. Statistical significant correlation was not seen between the presence of microsporidia and age, gender, stool appearance, and geographical region. Molecular analysis showed that all E. bieneusi were the genotype D. Phylogenetic tree demonstrated no classification according to the presence/absence of immunodeficiency, geographical locations and presence of diarrhea. The high prevalence of Encephalitozoon sp., in comparison to E. bieneusi in this study suggested the importance of this genus alongside with E. bieneusi in Iran. In addition, predominance of the genotype D highlighted the wide distribution of this genotype in Iran.


Assuntos
Encephalitozoon , Enterocytozoon , Microsporidiose/epidemiologia , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , DNA Espaçador Ribossômico , Encephalitozoon/classificação , Encephalitozoon/genética , Encephalitozoon/isolamento & purificação , Enterocytozoon/classificação , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , Feminino , Genes Fúngicos , Humanos , Hospedeiro Imunocomprometido , Irã (Geográfico)/epidemiologia , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Infecções Oportunistas/epidemiologia , Patologia Molecular/métodos , Filogenia , RNA Ribossômico 18S , Adulto Jovem
16.
Med Mycol ; 58(6): 810-819, 2020 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31868212

RESUMO

Despite the fact that many approaches have been developed over years to find efficient and well-tolerated therapeutic regimens for microsporidiosis, the effectiveness of current drugs remains doubtful, and effective drugs against specific targets are still scarce. The present study is the first that was designed to evaluate the potency of auranofin, an anti-rheumatoid FDA approved drug, against intestinal Encephalitozoon intestinalis. Evaluation of the drug was achieved through counting of fecal and intestinal spores, studying the intestinal histopathological changes, measuring of intestinal hydrogen peroxide level, and post therapy follow-up of mice for 2 weeks for detection of relapse. Results showed that auranofin has promising anti-microsporidia potential. It showed a promising efficacy in mice experimentally infected with E. intestinalis. It has revealed an obvious reduction in fecal spore shedding and intestinal tissue spore load, amelioration of intestinal tissue pathological changes, and improvement of the local inflammatory infiltration without significant changes in hydrogen peroxide level. Interestingly, auranofin prevented the relapse of infection. Thus, considering the results of the present work, auranofin could be considered a therapeutic alternative for the gold standard drug 'albendazole' against the intestinal E. intestinalis infection especially in relapsing cases.


Assuntos
Antifúngicos/uso terapêutico , Auranofina/uso terapêutico , Encephalitozoon/efeitos dos fármacos , Encefalitozoonose/tratamento farmacológico , Intestinos/microbiologia , Animais , Antifúngicos/farmacologia , Auranofina/farmacologia , Modelos Animais de Doenças , Encephalitozoon/genética , Encefalitozoonose/microbiologia , Fezes/microbiologia , Humanos , Masculino , Camundongos , Microsporídios/efeitos dos fármacos , Microsporidiose/tratamento farmacológico , Microsporidiose/microbiologia , Prevenção Secundária
17.
mBio ; 10(4)2019 08 20.
Artigo em Inglês | MEDLINE | ID: mdl-31431557

RESUMO

Microsporidia are opportunistic intracellular pathogens that can infect a wide variety of hosts ranging from invertebrates to vertebrates. During invasion, the microsporidian polar tube pushes into the host cell, creating a protective microenvironment, the invasion synapse, into which the sporoplasm extrudes. Within the synapse, the sporoplasm then invades the host cell, forming a parasitophorous vacuole (PV). Using a proteomic approach, we identified Encephalitozoon hellem sporoplasm surface protein 1 (EhSSP1), which localized to the surface of extruded sporoplasms. EhSSP1 was also found to interact with polar tube protein 4 (PTP4). Recombinant EhSSP1 (rEhSSP1) bound to human foreskin fibroblasts, and both anti-EhSSP1 and rEhSSP1 caused decreased levels of host cell invasion, suggesting that interaction of SSP1 with the host cell was involved in invasion. Coimmunoprecipitation (Co-IP) followed by proteomic analysis identified host cell voltage-dependent anion channels (VDACs) as EhSSP1 interacting proteins. Yeast two-hybrid assays demonstrated that EhSSP1 was able to interact with VDAC1, VDAC2, and VDAC3. rEhSSP1 colocalized with the host mitochondria which were associated with microsporidian PVs in infected cells. Transmission electron microscopy revealed that the outer mitochondrial membrane interacted with meronts and the PV membrane, mitochondria clustered around meronts, and the VDACs were concentrated at the interface of mitochondria and parasite. Knockdown of VDAC1, VDAC2, and VDAC3 in host cells resulted in significant decreases in the number and size of the PVs and a decrease in mitochondrial PV association. The interaction of EhSSP1 with VDAC probably plays an important part in energy acquisition by microsporidia via its role in the association of mitochondria with the PV.IMPORTANCE Microsporidia are important opportunistic human pathogens in immune-suppressed individuals, such as those with HIV/AIDS and recipients of organ transplants. The sporoplasm is critical for establishing microsporidian infection. Despite the biological importance of this structure for transmission, there is limited information about its structure and composition that could be targeted for therapeutic intervention. Here, we identified a novel E. hellem sporoplasm surface protein, EhSSP1, and demonstrated that it can bind to host cell mitochondria via host VDAC. Our data strongly suggest that the interaction between SSP1 and VDAC is important for the association of mitochondria with the parasitophorous vacuole during microsporidian infection. In addition, binding of SSP1 to the host cell is associated with the final steps of invasion in the invasion synapse.


Assuntos
Interações Hospedeiro-Patógeno/fisiologia , Microsporídios/metabolismo , Mitocôndrias/microbiologia , Canais de Ânion Dependentes de Voltagem/metabolismo , Citoplasma , Encephalitozoon , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Técnicas de Silenciamento de Genes , Inativação Gênica , Humanos , Mitocôndrias/metabolismo , Proteínas de Transporte da Membrana Mitocondrial/genética , Proteômica , Canal de Ânion 1 Dependente de Voltagem/genética , Canal de Ânion 2 Dependente de Voltagem/genética , Canais de Ânion Dependentes de Voltagem/genética
18.
Diagn Microbiol Infect Dis ; 95(1): 34-37, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31079868

RESUMO

Cryptosporidium spp. Enterocytozoon bieneusi and Encephalitozoon intestinalis are opportunistic pathogens responsible for gastrointestinal diseases. We evaluated the ParaGENIE Crypto-Micro Real-Time PCR kit (Ademtech, France), the first CE-IVD compliant PCR assay available for these pathogens. This study was conducted blindly against a reference panel of 115 stool specimens including positive samples for Cryptosporidium spp. (n = 48) and E. bieneusi (n = 38) as well as negative or positive samples for other parasites to test for cross-reactivity. An additional set of samples corresponding to 8 rare Cryptosporidium species was also included. Discrepancies were evaluated with external in-house PCR tests. The ParaGENIE Crypto-Micro PCR assay displayed a sensitivity/specificity of 91.7%/100% and 97.3%/98.7% for Cryptosporidium spp. and E. bieneusi, respectively, and was able to detect all 12 Cryptosporidium species of the reference panel, including rare species. This new CE-IVD assay will facilitate the diagnosis of intestinal cryptosporidiosis and microsporidiosis, a major concern in immunocompromised patients and travelers.


Assuntos
Técnicas Bacteriológicas/métodos , Cryptosporidium/genética , Encephalitozoon/genética , Enterocytozoon/genética , Fezes/microbiologia , Fezes/parasitologia , Reação em Cadeia da Polimerase Multiplex , Cryptosporidium/classificação , DNA Fúngico/genética , DNA de Protozoário/genética , Encephalitozoon/classificação , Enterocytozoon/classificação , Humanos , Kit de Reagentes para Diagnóstico , Sensibilidade e Especificidade
19.
J Eukaryot Microbiol ; 66(5): 771-777, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-30849216

RESUMO

Microsporidia are opportunistic pathogens that infect a wide range of invertebrates and vertebrates. To assess the potential role of dogs in the transmission of these zoonotic pathogens, a total of 282 fecal samples from dogs in the Central Anatolia Region of Turkey were analyzed by utilizing species specific polymerase chain reaction for the four most frequent human microsporidia. Two microsporidia species were recognized in 41 samples (14.5%). Encephalitozoon intestinalis was detected in 35 samples (12.4%) and it was the most common microsporidium. The second microsporidium, E. cuniculi, was identified in six (2.1%) of the samples. Sequence analysis of the intergenic spacer of the ribosomal ribonucleic acid (RNA) internal transcribed spacer (ITS) gene revealed the presence of three E. intestinalis haplotypes closely associated with each other. No polymorphic region was found among the ITS sequences of E. cuniculi isolates and they were characterized as genotype III. This study provides the first data on the zoonotic microsporidia species from dogs in Turkey.


Assuntos
Doenças do Cão/microbiologia , Encephalitozoon/isolamento & purificação , Encefalitozoonose/microbiologia , Microsporídios/isolamento & purificação , Microsporidiose/microbiologia , Animais , Doenças do Cão/epidemiologia , Cães , Encephalitozoon/classificação , Encephalitozoon/genética , Encefalitozoonose/epidemiologia , Fezes/microbiologia , Genótipo , Humanos , Microsporídios/classificação , Microsporídios/genética , Microsporidiose/epidemiologia , Turquia/epidemiologia , Zoonoses/microbiologia , Zoonoses/transmissão
20.
J Vet Diagn Invest ; 31(3): 467-470, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30795726

RESUMO

Two male juvenile central bearded dragons ( Pogona vitticeps) were submitted for postmortem examination after dying at their respective homes. Dragon 1 had marked hemopericardium with restrictive epicarditis. The inner aspect of the distended pericardial sac was lined by a fibrinoheterophilic membrane. In addition, granulomas abutted the testes. Dragon 2 had acute hemopericardium and granulomatous arteritis of the great vessels exiting the heart. Histologically, both animals had granulomatous arteritis of the large arteries with intrahistiocytic gram-positive, slightly elongated, up to 2 µm long microorganisms that contained a vacuole. These microorganisms were also present in the paratesticular granulomas. On transmission electron microscopy, the microorganisms were identified as microsporidians given the presence of exospore, endospore, vacuole, nucleus, and a filament with 4-6 coils. The microsporidia were identified as Encephalitozoon pogonae based on sequencing of the internal transcribed spacer 1 of the ribosomal RNA genes. Microsporidia are agents of disease in bearded dragons. Intrapericardial arteritis of large arteries with hemopericardium or restrictive epicarditis is a fatal manifestation of this infection.


Assuntos
Arterite/veterinária , Encephalitozoon/isolamento & purificação , Encefalitozoonose/veterinária , Lagartos , Derrame Pericárdico/veterinária , Animais , Arterite/microbiologia , Arterite/patologia , Encephalitozoon/genética , Encephalitozoon/ultraestrutura , Encefalitozoonose/microbiologia , Encefalitozoonose/patologia , Evolução Fatal , Masculino , Microscopia Eletrônica de Transmissão , Derrame Pericárdico/microbiologia , Derrame Pericárdico/patologia
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