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A 27-year-old immunocompetent woman presented to our clinic with redness, mild pain and blurring of vision in her left eye (LE) for 3 weeks with no preceding history of injury or contact lens wear. At presentation, her visual acuity was 20/20 and 20/60 in the right eye and LE, respectively. Slit

Microsporidial stromal keratitis is an increasingly well-known vision-threatening disease. A large proportion of cases are initially misdiagnosed as herpes simplex keratitis and treated with topical steroids. In most of such cases, medical treatment failed, and corneal transplantation was required. This study reported the results of 0.02% topical chlorhexidine used to treat three cases of microsporidial stromal keratitis and reviewed the literature on the outcomes of microsporidial stromal keratitis treatment. In the first case, histopathology of a specimen from penetrating keratoplasty (PK) revealed severe chronic inflammation involving the entire stromal layer but no microorganism activity after the application of topical chlorhexidine for 10 months. The second case exhibited complete resolution of keratitis after topical chlorhexidine. The patient in the third case did not respond to medical treatment, and therapeutic PK was performed. Histopathological examination revealed numerous microsporidial spores that had colonized in the mid and deep stroma, where few inflammatory cells were observed. These findings explain the variable microsporidial susceptibility to chlorhexidine, suggesting the crucial role of host immunity. In cases of host immunity, topical chlorhexidine may represent a promising option for the treatment of microsporidial stromal keratitis.

Background Microsporidia are rarely reported to cause outbreaks of diarrhea. We describe a foodborne outbreak of microsporidiosis from a workplace canteen in November 2020 in Denmark. Methods A probable case was defined as any person using the canteen between 4 November and 13 December 2020, reporting at least one gastrointestinal symptom, whereas a confirmed case also had an Enterocytozoon bieneusi positive stool sample. A web-based questionnaire was used to collect clinical, epidemiological, and food exposure data. We performed a retrospective cohort study and tested stool samples from affected individuals for bacterial, viral, and parasitic pathogens, including E. bieneusi. Results Altogether, 195 individuals completed the questionnaire. We identified 52 cases (65% male; median age 45 years [range 25–65]). Diarrhea (90%), fatigue (83%), and abdominal pain (79%) were the most commonly reported symptoms. Eight cases were laboratory-confirmed and had E. bieneusi genotype C. The incubation period was between 5 and 12 days, and polymerase chain reaction (PCR)-detectable spore shedding occurred up to 43 days after symptom onset. Disease was associated with consuming food from the workplace canteen on 4 November 2020 (relative risk [RR[, 2.8 [95% confidence interval [CI]: 1.4 – 5.4]) and lunchboxes containing open sandwiches (RR, 3.2 [95% CI: 1.4 – 7.2]) served that day. Conclusions This is the second documented foodborne outbreak of E. bieneusi genotype C-associated diarrhea worldwide. Epidemiological findings advocated an open sandwiches lunchbox from 4 November 2020, as a likely source. E. bieneusi may be an under-reported cause of outbreaks of diarrhea, and testing for it might be useful in foodborne outbreak investigations.

Microsporidial myositis caused by Trachipleistophora hominis is a life-threatening and emerging microsporidiosis among immunocompromised hosts. This article reports a case of disseminated microsporidiosis caused by T. hominis in southern Thailand. The patient had HIV and presented at the clinic with incapacitating muscle pain. She was diagnosed with disseminated microsporidiosis. Molecular identification revealed the sequence of 18S ribosomal RNA gene involving sequences sharing 99% nucleotide identity with T. hominis from an Australian patient. To our knowledge, this is the first study to report the detection of T. hominis microsporidia in an HIV patient in Thailand.

Gastrointestinal microsporidiosis is a major cause of chronic diarrhea in people with acquired immune deficienc syndrome. However, it can also affect individuals with inherited or congenital immunodeficiency. We reported a case of intestinal microsporidiosis and salmonellosis in a young boy with Hyper IgM syndrome. He presented with severe diarrhea and fever for 5 days. Stool examination showed heavy infection of Microsporidia spp. PCR confirmed the species as Enterocytozoon bieneusi. He was treated with albendazole for 25 days which abated the symptoms. To our knowledge, this is the first reported case of intestinal microsporidiosis in inherited immunodeficiency, X-linked Hyper IgM (XHIGM).

Background Microsporidia is a zoonotic pathogen with health consequences in immunocompromised patients. Small ruminants are a potential reservoir of microsporidia for humans in their vicinity. Hence, we aimed to evaluate the molecular prevalence of microsporidian infections with emphasis on Enterocytozoon bieneusi genotypes among sheep and goats at a global scale through systematic review and meta-analysis approach. Methods The standard protocol of preferred reporting items for systematic reviews and meta-analyses (PRISMA) guidelines were followed. Eligible prevalence studies on small ruminant microsporidiosis, published from 1 January 2000 until 15 April 2021 were gathered using systematic literature search in PubMed, Scopus, Web of Science and Google Scholar databases. Inclusion and exclusion criteria were applied. The point estimates and 95% confidence intervals were calculated using a random-effects model. The variance between studies (heterogeneity) was quantified by I2 index. Results In total, 25 articles (including 34 datasets) were included for final meta-analysis. The pooled molecular prevalence of microsporidia in sheep and goats was estimated to be 17.4% (95% CI: 11.8–25%) and 16% (95% CI: 11.2–22.4%), respectively. Likewise, the overall prevalence of E. bieneusi was estimated to be 17.4% (95% CI: 11.8–25%) for sheep and 16.3% (95% CI: 11.3–22.8%) for goats. According to internal transcribed spacer (ITS) gene analysis, E. bieneusi with genotypes BEB6 (15 studies) and COS-1 (nine studies) in sheep, and CHG3 (six studies) and BEB6 (five studies) in goats were the highest reported genotypes. Conclusion The present results highlight the role of sheep and goats as reservoir hosts for human-infecting microsporidia. Therefore, this global estimate could be beneficial on preventive and control measures.

Enterocytozoon bieneusi is a widespread opportunistic pathogen found in humans and domestic animals, including cattle that poses a public health risk. This study was performed to evaluate the prevalence, genotypic diversity, and zoonotic potential of E. bieneusi among children and calves in Bangladesh. A total of 998 fecal samples were collected from children (n = 299) and calves (n = 699) and screened by nested PCR and sequencing of the ribosomal internal transcribed spacer (ITS). The overall prevalence of E. bieneusi infection was 6.4% in children and 7.9% in calves. ITS sequence analysis of 74 isolates revealed 10 genotypes, including eight known genotypes (A, D, Type IV, PigEBITS7, I, J, BEB4, and BEB6) and two new genotypes (BANEB1 and BANEB3). Specifically, genotypes A, D, Type IV, PigEBITS7, BANEB1, and BANEB3, and genotypes D, PigEBITS7, I, J, BEB4, and BEB6 were detected in children and calves, respectively. Among them, genotypes D and I were dominant genotypes in children and calves, respectively. The genotypes D and PigEBITS7 were found in both children and calves, with PigEBITS7 being observed for the first time in calves. In phylogenetic analysis, six genotypes (A, D, Type IV, PigEBITS7, BANEB1, and BANEB3), detected in 39.2% of the isolates, belonged to zoonotic Group 1. The remaining four genotypes I, J, BEB4, and BEB6 were clustered in Group 2 and are common members of the group with zoonotic potential. To the best of our knowledge, this study provides the first report of E. bieneusi infection in calves in Bangladesh and also the first molecular characterization of the parasite in children and calves in this country. Two new genotypes in children have been found, which is noteworthy. Furthermore, the presence of zoonotic genotypes indicates that cattle may serve as reservoirs for E. bieneusi, which can be a source of human microsporidiosis.

Arctic char Salvelinus alpinus is an important cultural and subsistence resource for Inuit communities. Muscular infections by microsporidia were diagnosed for the first time in Arctic char originating from 2 different lakes in Nunavik (Quebec, Canada). The consumption of these infected fish was associated with digestive tract disorders in people. To better characterize microsporidiosis in these char populations, a cross-sectional study was conducted on 91 fish. The microsporidium was classified as a member of the Microsporidium collective genus by morphological evaluation and phylogenetic analysis using small subunit ribosomal DNA sequence data. The presence and severity of infection were determined histologically. Microsporidian infection occurred in 61% of the fish (56/91) and was significantly associated with an increase in their age, length and weight. The severity of infection (percentage of muscle area affected by microsporidia) was mild in most cases (

Background and objective: To describe the clinical course and propose a morphological classification scheme of microsporidial keratoconjunctivitis. Methods: This is a prospective study at L V Prasa...

CASE REPORT A 1-year-old, neutered male German Shepherd was presented with a 5-month history of episodic lethargy, intermittent fever, weight loss and a hunched posture. The dog was diagnosed with presumptive microsporidian meningoencephalitis based on cytological findings on cerebrospinal fluid analysis and a positive PCR test. The dog initially responded favourably to a 4-week course of trimethoprim-sulfadiazine, pyrimethamine and fenbendazole, and remained well for 12 weeks following cessation of treatment. Disease then recurred, and despite an initial positive response to treatment, he deteriorated and was euthanased 11 weeks later, 7.5 months after definitive diagnosis and 13 months after clinical signs were first reported. CONCLUSION To the authors knowledge, this is the first case of canine microsporidiosis in Australia.

Microsporidia are obligate intracellular fungi with a remarkable ability to infect a wide range of invertebrate and vertebrate hosts. Namely, Enterocytozoon bieneusi is the most frequently microsporidia reported worldwide, and mainly associated with chronic diarrhea and wasting syndrome in AIDS patients. Microscopy and PCR-based detection techniques are effective for diagnosis and identification of species and genotypes; however, these methods should be standardized in each laboratory. In this study, we performed microscopy and nested PCR techniques with PCR product sequencing to detect E. bieneusi in human stool samples. These techniques, if applied together, might prove useful for diagnosis and future epidemiological studies of intestinal microsporidiosis in Argentina.

After cardiovascular diseases, infectious diseases are the second most common cause of death worldwide. Although these infections are caused mainly by viruses or bacteria, a systematically growing prevalence of human and animal opportunistic fungal infections is noticeable worldwide. More attention is being paid to this problem, especially due to the growing frequency of recalcitrant and recurrent mycoses. The latter are classically divided into superficial, which are the most common type, subcutaneous, and systemic. This work discusses opportunistic fungal pathogens without proven horizontal transmission between different animal species including humans and microsporidia as spore-forming unicellular parasites related to fungi; however, with a yet undetermined taxonomic position. The review also mentions aetiological agents, risk factors, epidemiology, geographical distribution, and finally symptoms characteristic for individual disease entities. This paper provides insight into fungal infections from a global perspective and simultaneously draws attention to emerging pathogens, whose prevalence is continuously increasing. Finally, this work also takes into consideration the correct nomenclature of fungal disease entities and the importance of secondary metabolites in the pathogenesis of fungal infections.

This study assessed intestinal protozoa with stress on Blastocystis among patients with CKD andthose on hemodialysis (HD) in Nephrology Unit, Medical Research Institute, Alexandria.The patients were divided into three groups. GI: 50 patients on HD for at least one year, GII: 50 CKDpatients not on HD, and GIII: 50 healthy controls. Fresh stool samples were collected in labeled plasticboxes and examined as direct smears after staining with trichrome, Modified Ziehl-Neelsen (MZN)stains and modified trichrome (MT) stains. Besides, stool samples were cultured for Blastocystis hominisand Copro-antigen (ICT) was used to detect E. histolytica /dispar, G. lamblia and C. parvum.The results showed that the parasitic infections were detected in 79% of the patients; 40% in HD and39% in CKD patients. The positive samples were (47%) detected by direct wet mount followed byiodine smear (43%) and then trichrome stain (42%). Microsporidiosis was detected in 55 (55%) HDand CKD patients using the MT stain. ICT agreed with microscopic examination in diagnosis ofE.histolytica/ dispar and G. lamblia and showed perfect agreement with MZN in C. parvum diagnosis.

Enterocytozoon bieneusi is an obligate intracellular parasite and the most common pathogen of microsporidiosis in humans and animals. In this study, a total of 198 fecal samples were collected from 11 captive populations of Rhesus macaque in Chinese zoos, to investigate the prevalence and analyze the zoonotic potential of E. bieneusi by genotype of the Internal Transcribed Spacer (ITS) gene on the rRNA of E. bieneusi via nested PCR. Results showed that the average infection rate of E. bieneusi in the 11 populations was 13.6%, and the highest infection rate was 56.5% in the population of Xinjiang Tianshan Zoo. Seven genotypes were identified including 2 known genotypes (D and CM1) and 5 novel genotypes (Mul1, Mul2, Mul3, Mul4 and Mul5). Phylogenetic analysis revealed that the novel genotypes Mul2, Mul3, Mul4 and Mul5 belonged to Group 1 showed the zoonotic potential．These findings extend the distribution of E. bieneusi genotypes and provide baseline data for controlling E. bieneusi infection.

Enterocytozoon bieneusi is an obligate intracellular fungus-like parasite with high genetic diversity among mammalian and avian hosts. Based on polymorphism analysis of the ribosomal internal transcribed spacer (ITS), nearly 500 genotypes were identified within E. bieneusi. Those genotypes form several genetic groups that exhibit phenotypic differences in host specificity and zoonotic potential and probably have varying public health implications. Some of the genotypes in Group 1 (e.g., D, EbpC, and Type IV) and Group 2 (e.g., BEB4, BEB6, I, and J) are the most common ones that infect a variety of hosts including humans and thus are of public health importance. By contrast, those genotypes in other genetic groups (Groups 3-11) are mostly restricted to the hosts from which they were originally isolated, which would have unknown or limited impacts on public health. Advances on diagnosis and molecular typing of E. bieneusi are introduced in this review. Genotype distribution pattern of E. bieneusi in major domestic animal groups (pigs, cattle, sheep, goats, cats, and dogs), the role of those animals in zoonotic transmission of microsporidiosis, and food and water as potential vehicles for transmission are interpreted here as well. This review highlights the importance of including more genetic or epidemiological data obtained in the same geographical areas and using more reliable genetic markers to analyze the actual extent of host specificity in E. bieneusi, for the purpose of fully appreciating zoonotic risks of those domestic animals in close contacts with men and enhancing our understanding of the modes of transmission.

Microsporidian spores are emerging opportunistic disease agents that are classified as worldwide diseases. In tropical areas, diagnosis of microsporidiosis is still neglected, while it may be associated with water and causes diarrhea. The evaluation of the distribution of microsporidian spores in groundwater of sub-urban areas of the Centre region of Cameroon has been our main objective. This study was conducted from August 2018 to August 2019. Microsporidian spores were identified by Weber’s stain and were observed using the Olympus CK2 inverted microscopy with immersion oil at objective 100 ×. The abundance of microsporidian spores according to species rang from classes is given as follow; 188 ([2.5–4] × [1.8–3])1, 247 ([4.5–6] × [3–5])2, 286 ([5–6] × [4–5])3, 190 ([7–8] × [4–6])4, 88 ([8–9] × [5–7])5, 44 ([9–10] × [3–4])6, 33 ([8–12] × [5–7])7, 120 ([7–11] × [4–8])8 μm with an average density of 1 ± 1 spore/10 mL of 256 samples. The density of spores can highlight epidemiologically study about microsporidia, and of the control of drinking water. The presence of human-pathogenic microsporidia in groundwater may suggest the presence of environmental reservoirs that include animals. Densities of spores were also higher during the short rainy season showing the implication of rain in the distribution of environmental spores. Statistical analysis shows that abundance of microsporidian spores increases with organic matter, turbidity and suspended solid and decreases with the temperature and total dissolved solid (P

We analyzed the peripheral blood lymphocyte subsets of cancer patients infected with intestinal parasites, with an aim to find out the relationship between the levels of different types of lymphocytes with the prognosis of patients. 201 cancer patients aged 18 and over were included. Stool samples of the patients were examined using native-lugol, trichrome, modified trichrome (Weber's Trichrome stain), and modified Ziehl-Neelsen staining methods. Microsporidia and Cryptosporidium parvum were investigated at the genus and species levels using PCR. Lymphocyte subsets were determined by flow cytometry in blood samples. One or more parasite species were detected in 115 (56.7%) patients. The most common parasite species were Microsporidia, Blastocystis and Entamoeba coli, respectively. The frequency of parasites was high in patients with low lymphocyte percentage, CD3(+) T cell and CD3(+) CD4(+) T (Th) cell levels in blood samples studied by flow cytometry. Microsporidia infection was significantly higher in patients with low lymphocyte percentage and Th cell levels. Similarly, C. parvum infection was found to be significantly higher in patients with low T lymphocyte percentage and Th cell level. Finally, Blastocystis infection was significantly higher in patients with low lymphocyte percentage and CD4/CD8 ratio higher than 1. The decrease in lymphocyte percentage, CD3(+) T cell and Th cell count, and low CD4/CD8 ratio in cancer patients increase the frequency of intestinal parasitic infections. Based on these results, lymphocyte subsets may help identify cancer patients at high risk of opportunistic parasites. We suggest that opportunistic parasitic infections affecting the clinical course of the disease should be considered by clinicians during the follow-up and treatment of patients.

Background Microsporidia of the genus Encephalitozoon are generally connected with severe infections with lethal outcome in immunodeficient hosts. In immunocompetent hosts, microsporidiosis typically establishes a balanced host-parasite relationship that produces minimal clinically overt disease. Although the alimentary tract represents one of the main primary target tissues, the mechanisms of reaching other tissues during systemic microsporidian infections remain unclear. Methods In the present study, we tested the relation between inflammation induction in immunocompetent and immunodeficient mice and the presence of spores of E. cuniculi genotype II in selected organs and in fecal specimens by using molecular and histology methods. Results We reported the positive connection between inflammation induction and the significant increase of E. cuniculi genotype II occurrence in inflammation foci in both immunocompetent BALB/c and immunodeficient severe combined immunodeficient (SCID) mice in the acute phase of infection and the re-activation of latent microsporidial infection following inflammation induction in immunocompetent mice. Conclusion The results imply possible involvement of immune cells serving as vehicles transporting E. cuniculi genotype II purposefully across the whole host body towards inflammation. With increasing number of records of infections, it is necessary to reconsider microsporidia as agents responsible for various pathologies. The elucidation of possible connection with pro-inflammatory immune responses represents an important challenge with consequences for human health and development of therapeutic strategies.

Background Microsporidiosis is an emerging zoonotic disease that is considered a global public health concern. Dogs are suggested as one of potential reservoirs for transmitting the microsporidia infection to humans. However, there is little data on distribution of microsporidia in dogs. The current study aimed to evaluate the global prevalence and genetic diversity of microsporidia infection among the dog population. Methods We searched four major databases for studies reporting the prevalence of microsporidia in dogs until 30 May 2020. A random-effects model was used to estimate the overall and the subgroup-pooled prevalence of microsporidia across studies. Result Finally, a total of 32 studies (including 37 datasets) from 17 countries were included in this meta-analysis. The overall prevalence (95% CI) of microsporidia infection was estimated at 23.1% (13.5–36.8%) using microscopic methods, 20.9% (14.6–29%) using serological methods, and 8.4% (6.1–11.5%) using molecular methods. Molecular methods showed that the highest number of reports was related to Enterocytozoon bieneusi with a pooled prevalence of 6.5% (4.9–8.7%). Considering E. bieneusi genotypes, most studies reported the PtEb IX (10 studies) and the D (eight studies) genotypes. Conclusion These results emphasize the role of a dog as a reservoir host for human-infecting microsporidia. In addition, monitoring programs for human-infecting microsporidia in animals with close contact to humans should be considered.

Little is known regarding the Enterocytozoon bieneusi genotypes in snakes worldwide. In the present study, a total of 273 fecal samples were collected from pet snakes in Beijing, China. They were then tested for the presence of E. bieneusi by PCR amplification of the internal transcribed spacer (ITS) gene. The overall infection rate of E. bieneusi was 4.4% (12/273), with the highest infection rate (20%, 1/5) of E. bieneusi was found in the Black rat snake (Pantherophis obsoletus), whereas no positive samples were detected from both Milk (0/22) and Coast garter snakes (0/2). Eight genotypes were identified, including four known genotypes: EbpA (n = 1), EbpC (n = 5), Henan-III (n = 1), and SHR1 (n = 1), and four novel genotypes: CRep-5 (n = 1), CRep-6 (n = 1), CRep-7 (n = 1), and CRep-8 (n = 1). Among them, EbpC (41.7%, 5/12) was the predominant genotype. Phylogenetic analysis showed that seven genotypes belonged to group 1, while genotype SHR1 belonged to group 2. Genotypes EbpA, EbpC, and Henan-III have been previously reported in humans. This suggests that pet snakes are a potential source of zoonotic microsporidiosis transmission in China.

Microsporidia is a phylum of obligate emergent intracellular protist-like fungi pathogens that infect a broad range of hosts including vertebrates and invertebrates. Enterocytozoon bieneusi is the most common cause of microsporidiosis in humans, affecting primarily immunosuppressed patients but also reported in immunocompetent individuals. Epidemiological information on the presence and molecular diversity of E. bieneusi in livestock and wildlife in Spain is limited. Therefore, the occurrence of this microsporidia was investigated in sympatric extensively reared Iberian pigs (n = 186) and free ranging wild boars (n = 142) in the province of Cordoba, Southern Spain. Forty-two Iberian pigs (22.6%) and three wild boars (2.1%) were found E. bieneusi positive by PCR. In Iberian pigs, occurrence of E. bieneusi was significantly higher in sows than in fattening pigs (31.6% vs. 11.4%; p = .001). Five genotypes were identified in Iberian pigs, four previously reported (EbpA, PigEb4, O, Pig HN-II) and a novel genotype (named PigSpEb1), while only two genotypes were identified in wild boars, EbpA and novel genotype PigSpEb1. All five genotypes identified belong to Group 1 suggesting zoonotic potential. This study constitutes the first report on the occurrence and molecular characterization of E. bieneusi in Iberian pigs and wild boars. The identification of two genotypes with zoonotic potential in sympatric Iberian pigs and wild boars suggests that E. bieneusi can be potentially transmitted between those two hosts, but also implies that they may act as natural sources of microsporidia infection to other hosts including humans.

Research conducted on model organisms may be biased due to undetected pathogen infections. Recently, screening studies discovered high prevalence of the microsporidium Pseudoloma neurophilia in zebrafish (Danio rerio) facilities. This spore-forming unicellular parasite aggregates in brain regions associated with motor function and anxiety, and despite its high occurrence little is known about how sub-clinical infection affects behaviour. Here, we assessed how P. neurophilia infection alters the zebrafish´s response to four commonly used neurobehavioral tests, namely: mirror biting, open field, light/dark preference and social preference, used to quantify aggression, exploration, anxiety, and sociability. Although sociability and aggression remained unaltered, infected hosts exhibited reduced activity, elevated rates of freezing behaviour, and sex-specific effects on exploration. These results indicate that caution is warranted in the interpretation of zebrafish behaviour, particularly since in most cases infection status is unknown. This highlights the importance of comprehensive monitoring procedures to detect sub-clinical infections in laboratory animals.

A new microsporidian species,Globosporidium parameciigen. nov., sp. nov., fromParamecium primaureliais described on the basis of morphology, fine structure, and SSU rRNA gene sequence. This is the first case of microsporidiosis inParameciumreported so far. All observed stages of the life cycle are monokaryotic. The parasites develop in the cytoplasm, at least some part of the population in endoplasmic reticulum and its derivates. Meronts divide by binary fission. Sporogonial plasmodium divides by rosette-like budding. Early sporoblasts demonstrate a well-developed exospore forming blister-like structures. Spores with distinctive spherical shape are dimorphic in size (3.7 ± 0.2 and 1.9 ± 0.2μm). Both types of spores are characterized by a thin endospore, a short isofilar polar tube making one incomplete coil, a bipartite polaroplast, and a large posterior vacuole. Experimental infection was successful for 5 of 10 tested strains of theParamecium aureliaspecies complex. All susceptible strains belong to closely relatedP. primaureliaandP. pentaureliaspecies. Phylogenetic analysis placed the new species in the Clade 4 of Microsporidia and revealed its close relationship toEuplotespora binucleata(a microsporidium from the ciliateEuplotes woodruffi), toHelmichia lacustrisandMrazekia macrocyclopis, microsporidia from aquatic invertebrates.

Background: Diagnosis of microsporidiosis is difficult due to its small size needing special stains and identification by an expert. Nowadays, application of polymerase chain reaction (PCR) made diagnosis more sensitive, specific and easier. Objectives: To update the prevalence of intestinal microsporidia and to clarify their genotype patterns in symptomatic and asymptomatic immune compromised and immune competent cases. Patients and Methods: Totally, 323 stool samples were collected and subdivided as 173 from immune compromised (group I) and 150 from immune competent (group II) individuals. Samples were examined for microsporidiosis by light microscopy smears stained by Weber's modified trichrome (WMT) and modified ZiehlNeelsen (MZN), as well as by nested and RFLP PCR techniques. Results: Microsporidial spores were microscopically detected in 45/323 (13.9%) individuals; 25/173 (14.5%) immune compromised and 20/150 (13.3%) immune competent cases. In the two groups, 25/45 (55.6%) were symptomatic complaining of diarrhea, abdominal pain, distension, mal-digestion and weight loss, with statistical significant difference (P

Microsporidia are a group of obligate intracellular parasites that naturally infect domestic and wild animals. Human microsporidiosis is an increasingly recognized multisystem opportunistic infection. The clinical manifestations are diverse with diarrhea being the most common presenting symptom. We present a 52-year-old woman with a history of amyopathic dermatomyositis complicated by interstitial lung disease managed with mycophenolate mofetil and hydroxychloroquine who presented with a 7-month history of recurrent subcutaneous nodules as well as intermittent diarrhea and chronic sinusitis. A punch biopsy showed superficial and deep lymphocytic and granulomatous dermatitis with focal necrosis. Tissue stains for microorganisms revealed oval 1 to 3 μm spores within the necrotic areas in multiple tissue stains. Additional studies at the Centers for Disease Control and Prevention confirmed cutaneous microsporidiosis. This case is one of very few confirmed examples of cutaneous microsporidiosis reported in the literature.

Microsporidia are a rare and commonly misdiagnosed cause of corneal infection, accounting for approximately 0.4% of cases of microbial keratitis in some populations. Ocular microsporidiosis most often presents as either microsporidial keratoconjunctivitis (MKC) or microsporidial stromal keratitis (MSK). Though these two clinical entities exhibit similar symptomology, they are distinguished from one another by the time course for disease progression, findings on slit-lamp examination, and response to medical therapy. This review summarizes the current literature on the etiology and clinical presentation of microsporidial infections of the cornea and highlights ongoing developments in available diagnostic modalities and treatment regimens.

Purpose: Microsporidium is a spore-forming intracellular parasite that affects a wide range of hosts including humans. The tumor necrosis factor alpha (TNF-α) plays a key role in the immunity to infection with microsporidia. Recently, the TNF-α antagonists have proven successful in treating variable autoimmune diseases. In the current study, we aimed to investigate the impact of using TNF-α antagonists as a therapeutic regimen in the prevalence of infections with microsporidia. Materials and Methods: Diarrheal patients with distinct autoimmune diseases (n = 100) were assigned to the study. Patients taking anti-TNF-α medications (n = 60) were allocated to Group 1A and those undergoing non-TNF-α inhibitor treatment (n = 40) to Group 1B. Furthermore, patients with diarrhea without autoimmune disorders (n = 20) were allocated as controls. Stool specimens, 3 per patient, were collected and microscopically examined for microsporidia spores. A microsporidia-specific stool polymerase chain reaction was used to confirm the microscopic findings. Results: Microsporidia infection was identified in 28.3% (17/60), 10% (4/40), and in 5% (1/20) of patients in Group 1A, Group 1B, and in the control group, respectively. Overall, infection was significantly high in cases compared to the controls and in patients receiving TNF-α antagonists compared to patients not given TNF-α inhibitors (P < 0.05). Finally, infection was significantly higher in cases treated with TNF-α antagonists for ≥2 months compared to cases treated for

Cryptosporidium spp. and Enterocytozoon bieneusi are two well-known protist pathogens which can result in diarrhea in humans and animals. To examine the occurrence and genetic characteristics of Cryptosporidium spp. and E. bieneusi in pet red squirrels (Sciurus vulgaris), 314 fecal specimens were collected from red squirrels from four pet shops and owners in Sichuan province, China. Cryptosporidium spp. and E. bieneusi were examined by nested PCR targeting the partial small subunit rRNA (SSU rRNA) gene and the ribosomal internal transcribed spacer (ITS) gene respectively. The infection rates were 8.6% (27/314) for Cryptosporidium spp. and 19.4% (61/314) for E. bieneusi. Five Cryptosporidium species/genotypes were identified by DNA sequence analysis: Cryptosporidium rat genotype II (n = 8), Cryptosporidium ferret genotype (n = 8), Cryptosporidium chipmunk genotype III (n = 5), Cryptosporidium rat genotype I (n = 4), and Cryptosporidium parvum (n = 2). Additionally, a total of five E. bieneusi genotypes were revealed, including three known genotypes (D, SCC-2, and SCC-3) and two novel genotypes (RS01 and RS02). Phylogenetic analysis revealed that genotype D fell into group 1, whereas the remaining genotypes clustered into group 10. To our knowledge, this is the first study to report Cryptosporidium spp. and E. bieneusi in pet red squirrels in China. Moreover, C. parvum and genotype D of E. bieneusi, previously identified in humans, were also found in red squirrels, suggesting that red squirrels may give rise to cryptosporidiosis and microsporidiosis in humans through zoonotic transmissions. These results provide preliminary reference data for monitoring Cryptosporidium spp. and E. bieneusi infections in pet red squirrels and humans.

Enterocytozoon bieneusi, a unicellular enteric microsporidian parasite, can infect humans and a wide range of animals throughout the world. Although E. bieneusi has been identified in many animals, there is no information regarding the genotypes of E. bieneusi in pet birds in China. Birds are important sources of emerging infectious diseases that affect humans, and immunosuppressed individuals can be exposed to potential zoonotic agents shed by birds. The aim of the present study was to determine the prevalence and genotypic diversity of E. bieneusi in pet birds, as well as assessed its zoonotic potential. A total of 387 fecal samples were collected from Psittaciformes (n = 295), Passeriformes (n = 67), and Galliformes (n = 16) from four pet markets in Sichuan province, Southwestern China. The overall prevalence of E. bieneusi in pet birds was 25.1% based on nested polymerase chain reaction analysis of the internal transcribed spacer (ITS) region of the ribosomal RNA (rRNA) gene (Psittaciformes, 21.7%; Passeriformes, 37.3%; Galliformes, 50.0%). Eight genotypes of E. bieneusi were identified, including five known genotypes (D, SC02, BEB6, CHB1, and MJ5) and three novel genotypes (SCB-I, SCB-II, and SCB-III). In phylogenetic analysis, genotypes D and SC02 and one novel genotype SCB-II were clustered within group 1, genotype BEB6 was classified within group 2, and the remaining genotypes (CHB1, MJ5, SCB-I, and SCB-III) clustered with group 10. To the best of our knowledge, this is the first report of E. bieneusi infection in pet birds in China. Genotypes D, SC02, and BEB6 that have been previously identified in humans, were found in pet birds in this study, suggesting that these pet birds can be a potential source of human microsporidiosis in China. Keywords: Enterocytozoon bieneusi, ITS, Pet bird, Zoonotic potential, China

A microsporidial keratopathy is described in two dogs. Both dogs presented with a unilateral stromal keratopathy characterized by multifocal coalescing opacities, and the diagnosis was made on histopathologic examination of keratectomy specimens. Transmission electron microscopy (TEM) on formalin-fixed, paraffin-embedded corneal tissue was performed in one dog, and the morphologic features were consistent with Nosema species infection. Both dogs were initially diagnosed and treated by superficial keratectomy. One dog received additional antifungal medication and underwent a penetrating keratoplasty following local recurrence two years later. No other systemic lesions attributable to the microsporidial infection were identified clinically. The clinical and diagnostic pathology findings, treatment, and follow-up are discussed.

We analyzed 2 batches of environmental samples after a microsporidial keratoconjunctivitis outbreak in Taiwan. Results indicated a transmission route from a parking lot to a foot washing pool to a swimming pool and suggested that accumulation of mud in the foot washing pool during the rainy season might be a risk factor.

Microsporidia are obligate intracellular pathogens identified ∼150 years ago as the cause of pébrine, an economically important infection in silkworms. There are about 220 genera and 1,700 species of microsporidia, which are classified based on their ultrastructural features, developmental cycle, host-parasite relationship, and molecular analysis. Phylogenetic analysis suggests that microsporidia are related to the fungi, being grouped with the Cryptomycota as a basal branch or sister group to the fungi. Microsporidia can be transmitted by food and water and are likely zoonotic, as they parasitize a wide range of invertebrate and vertebrate hosts. Infection in humans occurs in both immunocompetent and immunodeficient hosts, e.g., in patients with organ transplantation, patients with advanced human immunodeficiency virus (HIV) infection, and patients receiving immune modulatory therapy such as anti-tumor necrosis factor alpha antibody. Clusters of infections due to latent infection in transplanted organs have also been demonstrated. Gastrointestinal infection is the most common manifestation; however, microsporidia can infect virtually any organ system, and infection has resulted in keratitis, myositis, cholecystitis, sinusitis, and encephalitis. Both albendazole and fumagillin have efficacy for the treatment of various species of microsporidia; however, albendazole has limited efficacy for the treatment of Enterocytozoon bieneusi. In addition, immune restoration can lead to resolution of infection. While the prevalence rate of microsporidiosis in patients with AIDS has fallen in the United States, due to the widespread use of combination antiretroviral therapy (cART), infection continues to occur throughout the world and is still seen in the United States in the setting of cART if a low CD4 count persists.

The aetiology of diarrhoea in a patient in Cuba with HIV was investigated. Although molecular diagnostics are still not used in many under-resourced settings, here traditional methods were supported by use of PCR. This approach enabled detection of a dual infection (Cystoisospora belli and Enterocytozoon bieneusi), the latter of which was not identified by microscopy with Didier's trichromic staining.

Simple Summary We here report the first detection of Enterocytozoon hepatopenaei (EHP) in cultured Pacific whiteleg shrimp (Litopenaeus vannamei) with a growth disorder in Korea. The histopathology, electron microscopy and DNA phylogeny indicated that the EHP is a unique strain first recorded in Korea. This finding highlights the need for closer monitoring and surveillance to control EHP in aquaculture to prevent disastrous economic losses. Abstract The consumption of cultured crustaceans has been steadily increasing, and Pacific whiteleg shrimp (Litopenaeus vannamei) are major cultivated invertebrates worldwide. However, shrimp productivity faces a variety of challenges, mainly related to outbreaks of lethal or growth retardation-related diseases. In particular, hepatopancreatic microsporidiosis caused by the microsporidian parasite Enterocytozoon hepatopenaei (EHP) is an important disease associated with growth retardation in shrimp. Here, we report the detection of EHP through histopathological, molecular and electron microscopy methods in the hepatopancreas of Pacific whiteleg shrimp with growth disorder in a South Korean farm. Phylogenetic analysis showed a clade distinct from the previously reported EHP strains isolated in Thailand, India, China and Vietnam. An EHP infection was not associated with inflammatory responses such as hemocyte infiltration. Although EHP infection has been reported worldwide, this is the first report in the shrimp aquaculture in Korea. Therefore, an EHP infection should be managed and monitored regularly for effective disease control and prevention.

Introduction Microsporidiosis is an emerging opportunistic infection in renal transplantation (RT) recipients. We aimed to describe its clinical presentation and treatment. Materials and methods We collected microsporidiosis cases identified in RT recipients between 2005 and 2019 in six French centers from the Crystal, Divat and Astre prospective databases. Results We report 68 RT recipients with intestinal microsporidiosis; the patients were predominantly male (61.8%), with a median age of 58 (46-69) years. Infection occurred at a median time of 3 (0.8-6.8) years posttransplant. Only Enterocytozoon bieneusi was found. Microsporidiosis manifested as diarrhea (98.5% of patients) with weight loss (72.1%) and acute renal injury (57.4%) without inflammatory biological parameters. The therapeutic approaches were no treatment (N = 9), reduction of the immunosuppressive regimen (∆IS) (N = 22), fumagillin alone (N = 9), fumagillin and ∆IS (N = 19), and albendazole or nitazoxanide and ∆IS (N = 9). Overall clinical remission was observed in 60 patients (88.2%). We observed no acute kidney rejection, renal transplant failure or death within 6 months after microsporidiosis. Conclusion E. bieneusi is an underestimated opportunistic pathogen in RT recipients, and infection with E. bieneusi leads to diarrhea with important dehydration and acute renal injury. The treatment is based on the reduction of the immunosuppressive regimen and the administration of fumagillin if available. This article is protected by copyright. All rights reserved.

Enterocytozoon bieneusi is a microsporidian and zoonotic species. This study investigated the prevalence and distribution of E. bieneusi genotypes in farmed masked palm civets using nested PCR, as well as assessed their zoonotic potential by phylogenetic analysis of the ITS region of the rRNA region. Here, we collected 251 fecal specimens from farmed masked palm civets (Paguma larvata) from the Hainan Island, China. In total, 128 of 251 samples were positive for E. bieneusi, with an average infection rate of 51.0%. Seventeen genotypes were identified including 12 known genotypes—HNR-VI (n = 56), SHR1 (n = 45), SHW7 (n = 6), KIN-1 (n = 3), D (n = 3), New1 (n = 3), EbpC (n = 2), CHC5 (n = 1), CHG19 (n = 1), CHN4 (n = 1), EbpA (n = 1), and Henan-III (n = 1)—and five novel genotypes (HNPL-I to HNPL-II; one each). Phylogenetic analysis categorized these genotypes into two groups. Thirteen of them were members of the zoonotic group 1, and the remaining four genotypes were in group 12. This study has shown that the infection rates of E. bieneusi in masked palm civets from Hainan were relatively high and provide baseline data to control and prevent microsporidiosis in farm-related communities. Therefore, infections in masked palm civets with zoonotic genotypes D, EbpC, CHN4, EbpA, KIN-1, and Henan-III should be considered potential threats to public health.

The microsporidia Nosema ceranae is an obligate intracellular parasite that causes honey bee mortality and contributes to colony collapse. Fumagillin is presently the only pharmacological control for N. ceranae infections in honey bees. Resistance is already emerging, and alternative controls are critically needed. Nosema spp. exhibit increased sensitivity to heat shock, a common proteotoxic stress. Thus, we hypothesized that targeting the Nosema proteasome, the major protease removing misfolded proteins, might be effective against N. ceranae infections in honey bees. Nosema genome analysis and molecular modeling revealed an unexpectedly compact proteasome apparently lacking multiple canonical subunits, but with highly conserved proteolytic active sites expected to be receptive to FDA-approved proteasome inhibitors. Indeed, N. ceranae were strikingly sensitive to pharmacological disruption of proteasome function at doses that were well tolerated by honey bees. Thus, proteasome inhibition is a novel candidate treatment strategy for microsporidia infection in honey bees.

A vertically transmitted microsporidian, Microsporidia MB, with the ability to disrupt Plasmodium development was reported in Anopheles arabiensis from Kenya, East Africa. To demonstrate its range of incidence, archived DNA samples from 7575 Anopheles mosquitoes collected from Ghana were screened. MB prevalence was observed at 1.8%. An. gambiae s.s constituted 87% of positive mosquitoes while the remaining were from An. coluzzii. Both sibling species had similar positivity rates (24% and 19%; p = 0.42) despite the significantly higher number of An. gambiae s.s analysed (An. gambiae s.s = 487; An. coluzzii = 94; p = 0.0005). The microsporidian was also more prevalent in emerged adults from field-collected larvae than field-caught adults (p Microsporidia MB in Anopheles mosquitoes in West Africa. It indicates possible widespread among malaria vector species and warrants investigations into the symbiont’s diversity across sub-Saharan Africa.

AIM To report a case of bilateral microsporidiosis with coexisting fungal infection in one eye. METHOD Retrospective interventional case report. RESULTS A 61-year-old man with uncontrolled diabetes presented with clinical and microbiological features of non-resolving fungal keratitis in the right eye since 3 months and underwent therapeutic penetrating keratoplasty (TPK) for the same. Fungal filaments along with oval bodies suspicious of microconidia were noted on calcofluor stain. A week following TPK, the patient presented with features of viral keratouveitis in the left eye which on microbiology was confirmed as microsporidiosis. Retrospectively, the right eye microbiology slides were reassessed, which confirmed the coexistence of fungus with microsporidiosis by acid-fast stain and polymerase chain reaction. CONCLUSION Structural resemblance of microconidia with microsporidial spores can be misleading, thus creating a need for awareness regarding the possible coexistence along with a need to suspect microsporidiosis in nonresponding clinically resembling viral keratitis.