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Turkiye Parazitolojii Dergisi Jun 2023In this study, we aimed to determine the prevalence of in healthy sheep in Van province using molecular techniques and to reveal genotypes of the detected isolates.
OBJECTIVE
In this study, we aimed to determine the prevalence of in healthy sheep in Van province using molecular techniques and to reveal genotypes of the detected isolates.
METHODS
A total of 200 healthy appearance sheep comprise 38 male and 162 female, 32 preweaned, 38 postweaned lamb and 130 adult sheep from several farms in the Van region were included in the study between May and September 2021. Genomic DNA (gDNA) extractions were utilized on fecal samples collected from sheep by commercial kits, and DNA was investigated by Nested polymerase chain reaction (PCR) amplifying ITS rRNA in the gDNA isolates. PCR products of the positive isolates were subjected to sequence analyze for genotyping and phylogenetic analyses of .
RESULTS
DNA was determined in 16 out of 200 examined sheep fecal gDNA samples (8.0%) by Nested PCR. The highest prevalence was determined in preweaned lambs with a rate of 18.8%. This was followed by postweaned lambs and adult sheep with a prevalence of 10.5% and 4.6%, respectively. The prevalence of the infection in males and females was 7.9% and 9.3%, respectively. All the ITS rRNA amplicons from 16 positive isolates were subjected to sequence analyses for genotyping and phylogenetic analyses. Sequence analyses revealed that all the isolates determined in sheep belonged to the BEB6 genotype and clustered in genogroup 2 of with the BEB6 isolates from different hosts in several countries.
CONCLUSION
Molecular epidemiological data on the prevalence of in sheep in Turkey were obtained with this study and the common genotype was determined as BEB6 in the research area. The obtained data contribute to the molecular epidemiology and diversity of in sheep.
Topics: Male; Animals; Sheep; Female; Enterocytozoon; Phylogeny; Prevalence; Microsporidiosis; Sheep Diseases; Genotype; Feces
PubMed: 37249107
DOI: 10.4274/tpd.galenos.2022.76476 -
Euro Surveillance : Bulletin Europeen... Aug 2023BackgroundEpidemics of keratoconjunctivitis may involve various aetiological agents. Microsporidia are an uncommon difficult-to-diagnose cause of such...
BackgroundEpidemics of keratoconjunctivitis may involve various aetiological agents. Microsporidia are an uncommon difficult-to-diagnose cause of such outbreaks.AimDuring the third quarter of 2022, a keratoconjunctivitis outbreak was reported across Israel, related to common water exposure to the Sea of Galilee. We report a comprehensive diagnostic approach that identified as the aetiology, serving as proof of concept for using real-time metagenomics for outbreak investigation.MethodsCorneal scraping samples from a clinical case were subjected to standard microbiological testing. Samples were tested by calcofluor white staining and metagenomic short-read sequencing. We analysed the metagenome for taxonomical assignment and isolation of metagenome-assembled genome (MAG). Targets for a novel PCR were identified, and the assay was applied to clinical and environmental samples and confirmed by long-read metagenomic sequencing.ResultsFluorescent microscopy was suggestive of microsporidiosis. The most abundant species (96.5%) on metagenomics analysis was . Annotation of the MAG confirmed the species assignment. A unique PCR target in the microsporidian rRNA gene was identified and validated against the clinical sample. The assay and metagenomic sequencing confirmed in an environmental sludge sample collected at the exposure site.ConclusionsThe real-time utilisation of metagenomics allowed species detection and development of diagnostic tools, which aided in outbreak source tracking and can be applied for future cases. Metagenomics allows a fully culture-independent investigation and is an important modality for public health microbiology.
Topics: Humans; Metagenome; Metagenomics; Israel; Keratoconjunctivitis; Microsporidia; Disease Outbreaks; High-Throughput Nucleotide Sequencing
PubMed: 37535472
DOI: 10.2807/1560-7917.ES.2023.28.31.2300010 -
BMC Infectious Diseases Nov 2023Most cases of microsporidial keratoconjunctivitis are found in the Southern hemisphere. Our purpose was to investigate the first outbreak of microsporidial... (Observational Study)
Observational Study
BACKGROUND
Most cases of microsporidial keratoconjunctivitis are found in the Southern hemisphere. Our purpose was to investigate the first outbreak of microsporidial keratoconjunctivitis in Japan among healthy, immunocompetent soccer players from the same team during a 1-month period.
CASE PRESENTATION
This study is an observational case series. The medical records were analyzed for five cases with microsporidial keratoconjunctivitis who presented within September 2022. All five cases were males between 28 and 36 years old. These previously healthy individuals belonged to the same football team. Their eyes were considered susceptible to contaminated water or dirt from the turf at game and practice sites. All cases involved unilateral conjunctivitis, with scattered round white lesions that showed positive fluorescein staining in the corneal epithelium. All cases experienced diminution of vision in the affected eye. In three cases, direct smears showed spores of approximately 2-3 μm in diameter. Polymerase chain reaction (PCR) analysis of corneal scrapes revealed partial amplification of microsporidial 18 S ribosomal RNA gene in four cases. Sequences of PCR products from all four cases showed 100% identity with strains of Vittaforma corneae previously reported from an outbreak in Singapore. All cases were treated with topical therapy, including voriconazole, fluorometholone, and levofloxacin. Four eyes underwent corneal scraping. After treatment, all eyes healed without residual opacities.
CONCLUSIONS
Only a few sporadic case reports of this disease have previously been reported in Japan. We detected V. corneae in our case series, representing what appears to be the first outbreak of microsporidial keratoconjunctivitis in Japan. Exposure to contaminated water or soil, in addition to inadequate sanitary facilities, represents a potential source of infection. Further investigations to clarify the characteristics of microsporidia seem warranted.
Topics: Male; Humans; Adult; Female; Microsporidiosis; Japan; Keratoconjunctivitis; Disease Outbreaks; Water
PubMed: 37915107
DOI: 10.1186/s12879-023-08767-y -
Journal of Laboratory Physicians Sep 2023Human microsporidiosis presents as an important and rapidly emerging opportunistic infection. However, the exact burden of this infection especially in the pediatric...
Human microsporidiosis presents as an important and rapidly emerging opportunistic infection. However, the exact burden of this infection especially in the pediatric population of Northern India remains unknown. In this study, we investigated the prevalence of microsporidia among human immunodeficiency virus (HIV)-positive and HIV-negative pediatric patients who presented with diarrhea. A total of 263 children were recruited consisting of 98 HIV seropositive with diarrhea and 165 HIV seronegative but with diarrhea. Morning stool samples were collected and both direct and formol ether concentrated samples were examined for the presence of intestinal parasites. The modified acid-fast staining was done for coccidian parasites and trichrome stain for microsporidia detection. Further, the species were detected using a real-time polymerase chain reaction (PCR) targeting a conserved region of the small ribosomal subunit rRNA gene of , , , and . Overall, one or more parasites were detected in 52.04% (51/98) of HIV seropositive and 53.33% (88/165) of seronegative children ( = 0.8391). However, coccidian parasites were detected in a significantly huge number of HIV seropositive children (21.43% [21/98]) as compared with HIV seronegative children (4.24% [7/165]). Microsporidial DNA could be detected in HIV seropositive with diarrhea children (17.35% [17/98]) by PCR. A significant correlation between low CD4 count (≤ 200/μL) and intestinal parasite positivity could be established. Microsporidia is a significant cause of diarrhea in HIV seropositive pediatric patients and should be kept in mind as one of the differential diagnoses in such patients.
PubMed: 37564235
DOI: 10.1055/s-0043-1764482 -
Emerging Infectious Diseases Jun 2024We retrospectively analyzed of 211 frozen cerebrospinal fluid samples from immunocompetent persons in the Czech Republic and detected 6 Encephalitozoon cuniculi-positive...
We retrospectively analyzed of 211 frozen cerebrospinal fluid samples from immunocompetent persons in the Czech Republic and detected 6 Encephalitozoon cuniculi-positive samples. Microsporidiosis is generally underestimated and patients are not usually tested for microsporidia, but latent infection in immunodeficient and immunocompetent patients can cause serious complications if not detected and treated.
Topics: Humans; Czech Republic; Encephalitozoon cuniculi; Encephalitozoonosis; Male; Female; Middle Aged; Adult; Retrospective Studies; Aged; Immunocompetence
PubMed: 38782145
DOI: 10.3201/eid3006.231585 -
Animals : An Open Access Journal From... Nov 2023(EHP) is highly contagious and can cause hepatopancreatic microsporidiosis (HPM), which is typically characterized by the slow growth of shrimp. In this study, the...
(EHP) is highly contagious and can cause hepatopancreatic microsporidiosis (HPM), which is typically characterized by the slow growth of shrimp. In this study, the differences in histology, metabolism, oxidative stress and growth between healthy and EHP-infected were analyzed using an EHP challenge experiment. Histology showed that EHP caused lesions in the hepatic tubules of , such as hepatic tubular atrophy and epithelial cell shedding, with mature spores. Meanwhile, white feces may appear when the infection is severe. Furthermore, the content of total protein, glycogen, ATP and glucose in the EHP challenge group was significantly reduced. The qPCR results showed that EHP infection changed the expression of key genes in glucose metabolism, among which hexokinase (), phosphofructokinase (), pyruvatekinase (), citrate synthase () and isocitric dehydrogenase () were significantly down-regulated, while phosphoenolpyruvate carboxykinase (), fructose bisphosphatase () and glucose-6-phosphatase () were significantly up-regulated. Obviously, the expression of growth-related genes was disordered. Simultaneously, the antioxidant genes manganese superoxide dismutase (), catalase (), glutathione peroxidase (), glutathione-S-transferases () and nuclear factor E2-related factor2 () were up-regulated to varying degrees in the EHP challenge group, and EHP infection induced significant increases in the oxidative damage products lipid peroxide (LPO) and malondialdehyde (MDA). Ultimately, the shrimp weight of the challenge group was 6.85 ± 0.86 g, which was significantly lower than that of the control group (8.95 ± 0.75 g). Taken together, we speculate that EHP changes the substance metabolism and growth process by causing oxidative damage to the hepatopancreas, which may lead to the growth retardation of .
PubMed: 38067012
DOI: 10.3390/ani13233661 -
Parasitology Apr 2024spp., and microsporidia are unicellular opportunistic pathogens that can cause gastrointestinal infections in both animals and humans. Since companion animals may...
spp., and microsporidia are unicellular opportunistic pathogens that can cause gastrointestinal infections in both animals and humans. Since companion animals may serve as a source of infection, the aim of the present screening study was to analyse the prevalence of these intestinal protists in fecal samples collected from dogs living in 10 animal shelters in central Europe (101 dogs from Poland and 86 from the Czech Republic), combined with molecular subtyping of the detected organisms in order to assess their genetic diversity. Genus-specific polymerase chain reactions were performed to detect DNA of the tested species and to conduct molecular subtyping in collected samples, followed by statistical evaluation of the data obtained (using or Fisher's tests). The observed prevalence was 15.5, 10.2, 1 and 1% for , , spp. and , respectively. Molecular evaluation has revealed the predominance of dog-specific genotypes ( XXe1 subtype; assemblages C and D; genotype II; genotypes D and PtEbIX), suggesting that shelter dogs do not pose a high risk of human transmission. Interestingly, the percentage distribution of the detected pathogens differed between both countries and individual shelters, suggesting that the risk of infection may be associated with conditions typical of a given location.
Topics: Animals; Dogs; Dog Diseases; Enterocytozoon; Cryptosporidium; Microsporidiosis; Poland; Cryptosporidiosis; Feces; Czech Republic; Giardiasis; Prevalence; Giardia; Genotype; Giardia lamblia; Host Specificity
PubMed: 38305092
DOI: 10.1017/S003118202400009X -
Food and Waterborne Parasitology Jun 2024is one of the most prevalent microsporidia species, responsible for more than 90% of human and animal microsporidiosis. Microsporidia species, particularly are...
is one of the most prevalent microsporidia species, responsible for more than 90% of human and animal microsporidiosis. Microsporidia species, particularly are frequently reported from waterborne and foodborne outbreaks. Therefore, early detection is crucial in clinics and outbreak investigations. This study aimed to design a loop-mediated isothermal amplification (LAMP) for rapid detection of Total DNA was extracted from 30 -positive samples, which had been confirmed with nested PCR. LAMP primers were designed based on the identical fragment of small subunit ribosomal RNA () gene. LAMP reactions were performed at 63 °C for 60 min. The sensitivity and specificity of the assay were analyzed and the results of amplification were compared to real-time PCR. Our results showed that the LAMP assay successfully amplified 25/30 (83.3%) samples. The specificity results indicated no false positive with other microorganisms. Furthermore, the LAMP method exhibited a sensitivity (limit of detection, LoD) as low as 34 ag/μL of total DNA. Compared to the LAMP assay, real-time PCR was able to detect all 30 nested PCR-positive samples. Our findings showed that the LAMP assay was able to detect 83.3% of positive samples. Although the current assay was not able to detect all nested PCR-positive samples, the lack of need for specific instruments, rapid processes, and high specificity makes LAMP assay a suitable tool for screening.
PubMed: 38523772
DOI: 10.1016/j.fawpar.2024.e00225 -
Parasitology International Jun 2024Blastocystis sp., Enterocytozoon bieneusi, and Giardia duodenalis are three common zoonotic intestinal parasites, and cattle are important hosts of these three...
Blastocystis sp., Enterocytozoon bieneusi, and Giardia duodenalis are three common zoonotic intestinal parasites, and cattle are important hosts of these three intestinal protozoa. In this study, 1632 fecal samples were collected from dairy farms in Heilongjiang Province, China, and screened for Blastocystis sp., E. bieneusi, and G. duodenalis using polymerase chain reaction. Of these, 149 (9.13%) were positive for three zoonotic pathogens, including 104 (6.40%), 22 (1.35%), and 23 (1.41%) for Blastocystis sp., E. bieneusi, and G. duodenalis, respectively. Based on partial SSU rRNA gene sequencing analysis, 104 positive samples of Blastocystis sp. were found, and a total of nine known subtypes were identified, including ST10 (61), ST3 (18), ST14 (6), ST26 (7), ST24 (3), ST25 (2), ST1 (2), ST5 (2), and ST21 (1). Among these, three subtypes (ST1, ST3, and ST5) were recognized as zoonotic subtypes, and two subtypes (ST10 and ST14) were specific to animals. All 23 Giardia duodenalis-positive samples belonged to assemblage E (n = 23) based on sequenced beta-giardin (bg) and triosephosphate isomerase (tpi) genes. Three known genotypes of E. bieneusi, namely J (n = 9), I (n = 6), and BEB4 (n = 7), were identified by sequence analysis of the internal transcriptional spacer region gene. Our study provides basic data for prevention and control in Heilongjiang Province; however, further research is required to better understand the prevalence and public health significance of these pathogens in the Heilongjiang region.
Topics: Animals; Cattle; Giardia lamblia; Giardiasis; Enterocytozoon; Microsporidiosis; China; Genotype; Feces; Prevalence; Cryptosporidium
PubMed: 38403046
DOI: 10.1016/j.parint.2024.102871 -
BMC Veterinary Research Feb 2024Enterocytozoon bieneusi is a zoonotic pathogen widely distributed in animals and humans. It can cause diarrhea and even death in immunocompromised hosts. Approximately...
BACKGROUND
Enterocytozoon bieneusi is a zoonotic pathogen widely distributed in animals and humans. It can cause diarrhea and even death in immunocompromised hosts. Approximately 800 internal transcribed spacer (ITS) genotypes have been identified in E. bieneusi. Farmed foxes and raccoon dogs are closely associated to humans and might be the reservoir of E. bieneusi which is known to have zoonotic potential. However, there are only a few studies about E. bieneusi genotype identification and epidemiological survey in foxes and raccoon dogs in Henan and Hebei province. Thus, the present study investigated the infection rates and genotypes of E. bieneusi in farmed foxes and raccoon dogs in the Henan and Hebei provinces.
RESULT
A total of 704 and 884 fecal specimens were collected from foxes and raccoon dogs, respectively. Nested PCR was conducted based on ITS of ribosomal RNA (rRNA), and then multilocus sequence typing (MLST) was conducted to analyze the genotypes. The result showed that infection rates of E. bieneusi in foxes and raccoon dogs were 18.32% and 5.54%, respectively. Ten E. bieneusi genotypes with zoonotic potential (NCF2, NCF3, D, EbpC, CHN-DC1, SCF2, CHN-F1, Type IV, BEB4, and BEB6) were identified in foxes and raccoon dogs. Totally 178 ITS-positive DNA specimens were identified from foxes and raccoon dogs and these specimens were then subjected to MLST analysis. In the MLST analysis, 12, 2, 7 and 8 genotypes were identified in at the mini-/ micro-satellite loci MS1, MS3, MS4 and MS7, respectively. A total of 14 multilocus genotypes were generated using ClustalX 2.1 software. Overall, the present study evaluated the infection of E. bieneusi in foxes and raccoon dogs in the Henan and Hebei province, and investigated the zoonotic potential of the E. bieneusi in foxes and raccoon dogs.
CONCLUSIONS
These findings expand the geographic distribution information of E. bieneusi' host in China and was helpful in preventing against the infection of E. bieneusi with zoonotic potential in foxes and raccoon dogs.
Topics: Humans; Animals; Multilocus Sequence Typing; Enterocytozoon; Foxes; Raccoon Dogs; Molecular Epidemiology; Microsporidiosis; Feces; Prevalence; Phylogeny; China; Genotype
PubMed: 38341563
DOI: 10.1186/s12917-024-03883-6