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Parasites & Vectors Nov 2017Microsporidia are intracellular obligate parasites traditionally associated with immunosuppressed patients; their detection in immunocompetent patients has increased,...
BACKGROUND
Microsporidia are intracellular obligate parasites traditionally associated with immunosuppressed patients; their detection in immunocompetent patients has increased, highlighting their possible importance as emerging pathogens. Detection of spores in stools, urine, body fluids and tissues is difficult and immunological techniques such as immunofluorescence have proved to be a useful and reliable tool in the diagnosis of human microsporidiosis. For this reason, we have produced and characterized monoclonal antibodies (MAbs) specific for Encephalitozoon intestinalis (the second most frequent microsporidian infecting humans), and other Encephalitozoon species, that can be used in different diagnostic techniques.
RESULTS
Seven MAbs were selected in accordance with their optical density (OD). Four (4C4, 2C2, 2E5 and 2H2) were isotype IgG2a; two (3A5 and 3C9) isotype IgG3, and one Mab, 1D7, IgM isotype. The selected monoclonal antibody-secreting hybridomas were characterized by indirect immunofluorescence antibody test (IFAT), enzyme-linked immunosorbent assay (ELISA), Western blot, immunoelectron microscopy (Immunogold) and in vitro cultures. The study by IFAT showed different behavior depending on the MAbs studied. The MAbs 4C4, 2C2, 2E5 and 2H2 showed reactivity against epitopes in the wall of the spore (exospore and endospore) epitopes located in Encephalitozoon sp. spores, whereas the MAbs 3A5, 1D7 and 3C9 showed reactivity against internal epitopes (cytoplasmic contents or sporoplasm) of E. intestinalis spores. All MAbs recognized the developing parasites in the in vitro cultures of E. intestinalis. Additionally, 59 formalin-fixed stool samples that had been previously analyzed were screened, with 26 (44%) presenting microsporidian spores (18 samples with E. intestinalis and 8 samples with Enterocytozoon bieneusi). Detection of microsporidian spores by microscopy was performed using Calcofluor stain, Modified Trichrome, Quick-Hot Gram Chromotrope, as well as IFAT using MAbs 4C4, 2C2, 2E5 and 2H2. The 4 MAbs tested clearly recognized the larger spores corresponding to E. intestinalis, but showed no reactivity with Enterocytozoon bieneusi spores. The mass spectrometry and proteomic study revealed that the Mabs 4C4, 2C2, 2E5 and 2H2 recognized the Spore Wall Protein 1 (SWP1) as the antigenic target.
CONCLUSIONS
The IFAT-positive MAbs exhibited excellent reactivity against spores and developmental stages, permitting their use in human and animal diagnosis. The epitopes recognized (exospore, endospore and cytoplasmic contents) by the different MAbs developed need further study, and may reveal potential targets for vaccine development, immunotherapy and chemotherapy.
Topics: Antibodies, Monoclonal; Blotting, Western; Encephalitozoon; Encephalitozoonosis; Enterocytozoon; Feces; Fluorescent Antibody Technique; Humans; Mass Spectrometry; Microscopy; Microsporidiosis; Proteomics; Spores, Fungal
PubMed: 29121996
DOI: 10.1186/s13071-017-2503-z -
Microbiology Spectrum Dec 2021Pallas's squirrel (Callosciurus erythraeus) was introduced in Japan in the 1930s and has since established itself in several areas across the country. Although wild...
Prevalence and Molecular Characterization of the Zoonotic Enteric Protozoans spp., Enterocytozoon bieneusi, and from Pallas's Squirrels (Callosciurus erythraeus) in Kanagawa Prefecture, Japan.
Pallas's squirrel (Callosciurus erythraeus) was introduced in Japan in the 1930s and has since established itself in several areas across the country. Although wild Sciuridae populations have been demonstrated to be potential reservoirs for zoonotic enteric protozoa, epidemiological studies of such pathogens in Japan are scarce. Here, we examined 423 fecal samples from Pallas's squirrels captured in Kanagawa Prefecture, Japan, using PCR and DNA sequencing to determine the occurrence of spp., Enterocytozoon bieneusi, and . The overall prevalence of spp., E. bieneusi, and was 4.3% (18/423 samples), 13.0% (55/423 samples), and 44.0% (186/423 samples), respectively. The prevalence of and E. bieneusi was significantly higher in spring (60.1% and 17.4%, respectively) than in winter (27.6% and 8.6%, respectively [0.01]). Sequence analysis of spp., targeting the partial small subunit ribosomal RNA gene (SSU rDNA), showed 100% identity (541/541 bp) to Cryptosporidium ubiquitum, and analysis of the gp60 gene showed 99.76% (833/835 bp) identity to C. ubiquitum subtype XIIh. The sequences of the ribosomal internal transcribed spacer region of E. bieneusi and the partial SSU rDNA of were identified as E. bieneusi genotype SCC-2 and subtype 4, respectively. This study confirmed the presence of C. ubiquitum, E. bieneusi, and in Pallas's squirrels in Kanagawa Prefecture. Because Pallas's squirrels inhabit urban areas, living close to humans, the species may serve as a potential source of infection in human populations. Pallas's squirrel is designated a "regulated organism" under the Invasive Alien Species Act in Japan, and municipal authorities are introducing control measures to reduce its populations. It has been suggested that wild mammals may play a role in contaminating the environment with zoonotic pathogens. The present study detected the enteric pathogens Cryptosporidium ubiquitum, Enterocytozoon bieneusi, and in the feces of Pallas's squirrels inhabiting Kanagawa Prefecture, Japan. These pathogens persist in the environment and contaminate soils and water, which may potentially infect humans. Because Pallas's squirrels in Kanagawa Prefecture are found in urban areas, where they are in close contact with human populations, continued monitoring of zoonotic diseases among squirrel populations will be important for evaluating the significance of wildlife in pathogen transmission.
Topics: Animals; Blastocystis; Blastocystis Infections; Cryptosporidiosis; Cryptosporidium; Enterocytozoon; Genes, Protozoan; Japan; Microsporidiosis; Prevalence; RNA, Ribosomal; Ribosome Subunits, Small; Sciuridae; Seasons
PubMed: 34730381
DOI: 10.1128/Spectrum.00990-21 -
BMC Microbiology Nov 2023Enterocytozoon bieneusi, Encephalitozoon spp., Cryptosporidium spp., and Giardia duodenalis (G. intestinalis) are enteric pathogens that cause diarrhea in pigs. This...
First identification and coinfection detection of Enterocytozoon bieneusi, Encephalitozoon spp., Cryptosporidium spp. and Giardia duodenalis in diarrheic pigs in Southwest China.
BACKGROUND
Enterocytozoon bieneusi, Encephalitozoon spp., Cryptosporidium spp., and Giardia duodenalis (G. intestinalis) are enteric pathogens that cause diarrhea in pigs. This study aimed to determine the prevalence of these enteric parasites and their coinfection with E. bieneusi in diarrheic pigs in Southwest China (Chongqing and Sichuan) using nested polymerase chain reaction (nPCR) based methods.
RESULTS
A total of 514 fecal samples were collected from diarrheic pigs from 14 pig farms in Chongqing (five farms) and Sichuan (nine farms) Provinces. The prevalence of Encephalitozoon spp., Cryptosporidium spp. and G. duodenalis was 16.14% (83/514), 0% (0/514), and 8.95% (46/514), respectively. Nested PCR revealed 305 mono-infections of E. bieneusi, six of E. cuniculi, two of E. hellem, and nine of G. duodenalis and 106 concurrent infections of E. bieneusi with the other enteric pathogens. No infections of E. intestinalis and Cryptosporidium species were detected. The highest coinfection was detected between E. bieneusi and E. cuniculi (10.5%, 54/514), followed by E. bieneusi and G. duodenalis (5.8%, 30/514) and E. bieneusi and E. hellem (2.9%, 15/514). E. bieneusi was the most frequently detected enteric pathogen, followed by E. cuniculi, G. duodenalis and E. hellem. There was a significant age-related difference in the prevalence of E. cuniculi in fattening pigs (χ = 15.266, df = 3, P = 0.002) and G. duodenalis in suckling pigs (χ = 11.92, df = 3, P = 0.008) compared with the other age groups. Sequence analysis of the ITS region of Encephalitozoon species showed two genotypes (II and III) for E. cuniculi and one (TURK1B) for E. hellem. Only G. duodenalis assemblage A was identified in all nested PCR-positive samples. E. bieneusi was found more often than other enteric pathogens.
CONCLUSIONS
This study showed that E. bieneusi, Encephalitozoon spp. [E. cuniculi and E. hellem] and G. duodenalis were common enteric parasites in diarrheic pigs in Chongqing and Sichuan Provinces. In case of both mono-infection and coinfection, E. bieneusi was the most common enteric pathogen in diarrheic pigs. Thus, it may be a significant cause of diarrhea in pigs. Precautions should be taken to prevent the spread of these enteric parasites.
Topics: Animals; Swine; Giardia lamblia; Giardiasis; Enterocytozoon; Cryptosporidiosis; Cryptosporidium; Encephalitozoon; Coinfection; Microsporidiosis; China; Genotype; Feces; Diarrhea
PubMed: 37951859
DOI: 10.1186/s12866-023-03070-x -
Parasite (Paris, France) Sep 2008Human microsporidian infections have emerged following the onset of the AIDS pandemic. Microsporidia are unicellular eukaryotic parasites that form spores. They are an... (Review)
Review
Human microsporidian infections have emerged following the onset of the AIDS pandemic. Microsporidia are unicellular eukaryotic parasites that form spores. They are an exceptionally diverse group of parasites that infect a wide range of eukaryotic cells in numerous invertebrate and vertebrate hosts. Of the 14 species newly described as pathogens in human, Enterocytozoon bieneusi, which causes gastrointestinal diseases, is the most common agent of human infections. In the past fifteen years, E. bieneusi was also identified in environmental sources, especially in surface water, as well as in wild, domestic and farm animals. These findings raised concerns for waterborne, foodborne and zoonotic transmission. Molecular analyses of the 243-bp internal Transcribed spacer-(ITS) of the rRNA gene have revealed a considerable genetic variation within E. bieneusi isolates of human and animal origins, supporting the potential for zoonotic transmission. The focus of this revue is to present and discuss recent advances in diagnosis and zoonotic potential of E. bieneusi infections.
Topics: AIDS-Related Opportunistic Infections; Animals; DNA, Ribosomal Spacer; Diagnosis, Differential; Enterocytozoon; Humans; Microsporidiosis; Molecular Epidemiology; Sequence Analysis, DNA; Species Specificity; Spores, Fungal; Zoonoses
PubMed: 18814706
DOI: 10.1051/parasite/2008153349 -
BMJ Case Reports Feb 2022We describe the diagnosis and successful management of a case of stromal microsporidiosis, an important emerging ocular disease. Stromal microsporidiosis is recalcitrant...
We describe the diagnosis and successful management of a case of stromal microsporidiosis, an important emerging ocular disease. Stromal microsporidiosis is recalcitrant and very often requires therapeutic keratoplasty for effective eradication. We successfully managed a steroid-treated case diagnosed only after 9 months, with a combination of polyhexamethyl biguanide 0.04%, chlorhexidine 0.04% and fluconazole 0.3% eye drops supplemented with tablet albendazole. However, complete resolution was achieved only after epithelial debridement. Toxicity due to the drugs was not noted. Diagnostic delays, steroid use and inappropriate therapy are commonly observed in stromal microsporidiosis. In spite of these potential disadvantages, our case responded well with complete eradication of the infection. The disease being fairly indolent and slowly progressive, medical therapy should be continued, in the absence of progression or other complications. Epithelial debridement may facilitate healing.
Topics: Albendazole; Chlorhexidine; Eye Infections, Fungal; Humans; Microsporidiosis
PubMed: 35165126
DOI: 10.1136/bcr-2021-246233 -
International Journal of Infectious... Jul 2021
Topics: Humans; Microsporidiosis; Organ Transplantation; Transplant Recipients
PubMed: 33940190
DOI: 10.1016/j.ijid.2021.04.084 -
Parasites & Vectors May 2019Enterocytozoon bieneusi and Giardia duodenalis are common human and animal pathogens. Studies have increasingly shown that non-human primates (NHPs) are common hosts of...
BACKGROUND
Enterocytozoon bieneusi and Giardia duodenalis are common human and animal pathogens. Studies have increasingly shown that non-human primates (NHPs) are common hosts of these two zoonotic parasites. However, few studies have explored the genetic diversity and public health potential of these pathogens in laboratory monkeys. In this study, we examined the genetic diversity of the two pathogens in crab-eating macaques (Macaca fascicularis) in a commercial facility in Hainan, China.
RESULTS
Enterocytozoon bieneusi and G. duodenalis were detected by PCR analysis in 461/1452 (31.7%) and 469/1452 (32.3%) fecal specimens from the animals, respectively. Significantly higher detection rates of E. bieneusi were detected in males (36.5%, 258/706) than in females (26.7%, 160/599; χ = 14.391, P = 0.0001), in animals with loose stools (41.4%, 151/365) than those with normal stool (28.5%, 310/1087; χ = 20.83, P < 0.0001), and in animals of over 3 years of age (38.6%, 135/350) than those of 1-3 years (29.6%, 326/1,102; χ = 9.90, P = 0.0016). For G. duodenalis, the detection rate in males (33.4%, 236/706) was higher than in females but not statistically significant (30.2%, 181/599; χ = 1.54, P = 0.2152), in monkeys with loose stools (41.1%, 150/365) than those with normal stools (29.3%, 319/1087; χ = 17.25, P < 0.0001), and in monkeys of 1-3 years of age (36.6%, 403/1102) than those over 3 years (18.9%, 66/350; χ = 38.11, P < 0.0001). Nine E. bieneusi genotypes were detected in this study by DNA sequence analysis of the internal transcribed spacer of the rRNA gene, namely Type IV (236/461), Peru8 (42/461), Pongo2 (27/461), Peru11 (12/461), D (4/461) and PigEbITS7 (1/461) previously seen in NHPs as well as humans, and CM1 (119/461), CM2 (17/461) and CM3 (3/461) that had been only detected in NHPs. DNA sequence analyses of the tpi, gdh and bg loci identified all G. duodenalis specimens as having assemblage B. Altogether, eight (4 known and 4 new), seven (6 known and 1 new) and seven (4 known and 3 new) subtypes were seen at the tpi, gdh and bg loci, leading to the detection of 53 multi-locus genotypes (MLG-B-hn01 to MLG-B-hn53). Most of them were genetically related to those previously seen in common Old-World monkeys.
CONCLUSIONS
Data from this study indicate a common occurrence of zoonotic genotypes of E. bieneusi and assemblage B of G. duodenalis in farmed crab-eating macaques in Hainan, China.
Topics: Animals; China; Enterocytozoon; Feces; Female; Genetic Variation; Genotype; Giardia lamblia; Giardiasis; Macaca fascicularis; Male; Microsporidiosis; Phylogeny; Prevalence; Public Health; Sequence Analysis, DNA; Zoonoses
PubMed: 31118092
DOI: 10.1186/s13071-019-3511-y -
BMJ Case Reports Apr 2021
Topics: Anterior Chamber; Cytomegalovirus Infections; Eye Infections, Viral; Humans; Iris; Microsporidiosis
PubMed: 33795265
DOI: 10.1136/bcr-2020-237490 -
Annals of Parasitology 2017Enterocytozoon bieneusi is the most commonly identified Microsporidia in humans and has also been detected worldwide in a large group of wild living and domestic... (Review)
Review
Enterocytozoon bieneusi is the most commonly identified Microsporidia in humans and has also been detected worldwide in a large group of wild living and domestic animals. The identification of E. bieneusi in wildlife has raised the question of the importance of animal reservoirs in the epidemiology of microsporidiosis and the implications of the infection with this pathogen in hosts. This review summarizes the available molecular data on the variety of E. bieneusi genotypes, both potentially zoonotic and host-specific isolated from wild living mammals and birds. In contrast to microsporidial infections of humans and domestic animals, wildlife deserves attention as a source of significant environmental reservoir of E. bieneusi.
Topics: Animals; Animals, Wild; Disease Reservoirs; Enterocytozoon; Humans; Microsporidiosis; Zoonoses
PubMed: 29396924
DOI: 10.17420/ap6304.113 -
Tuberkuloz Ve Toraks Mar 2016Microsporidia spp. are ubiquitous and infect a wide variety of intervertebrates and vertebrates, including humans. Pulmonary microsporidiosis, characterized by...
INTRODUCTION
Microsporidia spp. are ubiquitous and infect a wide variety of intervertebrates and vertebrates, including humans. Pulmonary microsporidiosis, characterized by nonspecific symptoms like fever, cough and dyspnea, is often overlooked in the differential diagnosis of pulmonary infections in immunsupressed patients. In this study, we aimed to determine the prevalence of pulmonary microsporidiosis in iatrogenically immunosuppressed patients and to evaluate the patient characteristics.
MATERIALS AND METHODS
Bronchoalveolar lavage (BAL) specimens from 63 iatrogenically immunosuppressed patients and 28 controls were examined with PCR. By using PMP1 and PMP2 common primers specifically designed for Enterocytozoon bieneusi, Encephalitozoon intestinalis, Encephalitozoon cuniculi and Encephalitozoon hellem small-subunit ribosomal DNA (SSU-rDNA) regions at 250-279 bp were amplified. In addition, PCR positive BAL specimens were examined with modified trichrome staining method for Microsporidia spores.
RESULT
Out of 63 immunosuppressed patients, nine (14.2%) had Microsporidia spp., but none of the control patients had Microsporidia spp. on PCR. This difference between two groups was statistically significant (χ² =4.439; p=0.035). On the other hand there was not a statistically significant relationship between PCR positivity and patient characteristics such as gender and age. Of nine patients with Microsporidia PCR positive, only one had spores of Microsporidia sp. Out of eight patients without spores, one had Mycobacterium tuberculosis, one patient had Klebsiella pneumoniae and five patients had Pneumocystis jirovecii DNA.
CONCLUSION
This is the first study to evaluate the pulmonary microsporidiosis in immunosupressed patients in Turkey. The results of the study indicated that Microsporidia spp. should be taken into account in the differential diagnosis of pulmonary infections in immunosupressed patients and it is important to use molecular methods such as PCR in the laboratory diagnosis of the causative agent.
Topics: Adolescent; Adult; Aged; Animals; Bronchoalveolar Lavage Fluid; Common Variable Immunodeficiency; DNA, Fungal; Enterocytozoon; Female; Follow-Up Studies; Humans; Immunosuppression Therapy; Lung Diseases, Fungal; Male; Microsporidiosis; Middle Aged; Polymerase Chain Reaction; Retrospective Studies; Turkey; Young Adult
PubMed: 27266280
DOI: 10.5578/tt.10207