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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. [ABSTRACT FROM AUTHOR]

Background: Microsporidiosis has been largely reported in patients with acquired immunodeficiency syndrome, but emerged as a cause of persistent diarrhea in solid organ transplant patients. Methods: Through the French Microsporidiosis Network and the Groupe français de recherche en greffe de foie, we collected all microsporidiosis cases identified in liver transplant patients between 1995 and 2020 in France. Results: We identified 24 liver transplant recipients with microsporidiosis. Sex ratio was balanced and median age was 58.8 (3.5‐83.5) years (there were 4 children). Microsporidiosis occurred at a median time of 3.9 (0.1‐18.9) years post‐transplant. Median duration of diarrhea before diagnosis was 22 days (12‐45). Therapeutic care included immunosuppressive therapy changes in 20 patients, as follows: stop cyclosporine or tacrolimus (n = 2), dose reduction of cyclosporine or tacrolimus (n = 12), stop MMF (n = 5), and dose reduction of corticosteroids (n = 1). In addition, 15 patients received specific therapy against microsporidiosis: fumagillin (n = 11) or albendazole (n = 4). Median duration of treatment was 14 days (8‐45 days). Finally, 7 patients had immunosuppressive treatment tapering only. Microsporidiosis was complicated by renal failure in 15 patients, requiring dialysis in one case. Two patients had infection relapse. No patient presented proven rejection within the 3 months after microsporidiosis. None of the patients died within the 3 months after microsporidiosis. Conclusions: Microsporidiosis is a very rare infection after liver transplantation but can induce severe dehydration and renal failure. Therefore, it must be systematically sought in any case of persistent diarrhea after first line screening of frequent infectious causes. [ABSTRACT FROM AUTHOR]

The recently discovered Anopheles symbiont, Microsporidia MB , has a strong malaria transmission-blocking phenotype in Anopheles arabiensis , the predominant Anopheles gambiae species complex member in many active transmission areas in eastern Africa. The ability of Microsporidia MB to block Plasmodium transmission together with vertical transmission and avirulence makes it a candidate for the development of a symbiont-based malaria transmission blocking strategy. We investigate the characteristics and efficiencies of Microsporidia MB transmission between An. arabiensis mosquitoes. We show that Microsporidia MB is not transmitted between larvae but is effectively transmitted horizontally between adult mosquitoes. Notably, Microsporidia MB was only found to be transmitted between male and female An. arabiensis , suggesting sexual horizontal transmission. In addition, Microsporidia MB cells were observed infecting the An. arabiensis ejaculatory duct. Female An. arabiensis that acquire Microsporidia MB horizontally are able to transmit the symbiont vertically to their offspring. We also investigate the possibility that Microsporidia MB can infect alternate hosts that live in the same habitats as their An. arabiensis hosts, but find no other non-anopheline hosts. Notably, Microsporidia MB infections were found in another primary malaria African vector, Anopheles funestus s.s. The finding that Microsporidia MB can be transmitted horizontally is relevant for the development of dissemination strategies to control malaria that are based on the targeted release of Microsporidia MB infected Anopheles mosquitoes. [ABSTRACT FROM AUTHOR]

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 < 0.05). These results show that microsporidia may be waterborne pathogens and that population may be exposed of health risk. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The model organism Caenorhabditis elegans is susceptible to infection by obligate intracellular pathogens, specifically microsporidia and viruses. These intracellular pathogens infect intestinal cells, or, for some microsporidia, epidermal cells. Strikingly, intestinal cell infections by viruses or microsporidia trigger a common transcriptional response, activated in part by the ZIP-1 transcription factor. Among the strongest activated genes in this response are ubiquitin-pathway members and members of the pals family, an intriguing gene family with cross-regulations of different members of genomic clusters. Some of the induced genes participate in host defense against the pathogens, for example through ubiquitin-mediated inhibition. Other mechanisms defend the host specifically against viral infections, including antiviral RNA interference and uridylation. These various immune responses are altered by environmental factors and by intraspecific genetic variation of the host. These pathogens were first isolated 15 years ago and much remains to be discovered using C. elegans genetics; also, other intracellular pathogens of C. elegans may yet to be discovered. • The model organism Caenorhabditis elegans is susceptible to infection by natural intracellular pathogens. • Intestinal infections by microsporidia and viruses trigger a common transcriptional immune response. • Immunity against intracellular infections includes ubiquitin-mediated pathways, viral RNA interference and uridylation. • Immunity varies among natural C. elegans isolates and with abiotic and biotic environmental parameters. • New intracellular pathogens of nematodes are still being discovered, such as the bacterium Bordetella atropi. [ABSTRACT FROM AUTHOR]

Microsporidiosis is a fungal infection that generally causes digestive disorders, especially in immunocompromised hosts. Over a 4-day period in January 2018, 3 patients with hematologic malignancies who were admitted to the hematology unit of a hospital in France received diagnoses of Enterocytozoon bieneusi microsporidiosis. This unusually high incidence was investigated by sequence analysis at the internal transcribed spacer rDNA locus and then by 3 microsatellites and 1 minisatellite for multilocus genotyping. The 3 isolates had many sequence similarities and belonged to a new genotype closely related to genotype C. In addition, multilocus genotyping showed high genetic distances with all the other strains collected from epidemiologically unrelated persons; none of these strains belonged to the new genotype. These data confirm the epidemiologic link among the 3 patients and support a common source of infection.

Abstract Background Eurycea sosorum (Barton Springs salamander) and Eurycea nana (San Macros salamander) are listed as endangered and threatened species, respectively, by the U.S. Fish and Wildlife Service (USFWS) with habitats restricted to small regions near Austin, Texas, USA. The conservation efforts with the Eurycea salamanders at the captive breeding program in San Marcos Aquatic Resources Center (SMARC), a USFWS facility, have seen an unexpected and increased mortality rate over the past few years. The clinical signs of sick or dead salamanders included erythema, tail loss, asymmetric gills or brachial loss, rhabdomyolysis, kyphosis, and behavior changes, suggesting that an infectious disease might be the culprit. This study aimed to identify the cause of the infection, determine the taxonomic position of the pathogen, and investigate the potential reservoirs of the pathogen in the environment. Results Histopathological examination indicated microsporidian infection (microsporidiosis) in the sick and dead Eurycea salamanders that was later confirmed by PCR detection. We also determined the near full-length small subunit ribosomal RNA (SSU rRNA) gene from the microsporidian pathogen, which allowed us to determine its phylogenetic position, and to design primers for specific and sensitive detection of the pathogen. Phylogenetic analysis indicated that this pathogen was closely related to the insect parasites Vavraia spp. and the human opportunistic pathogen, Trachipleistophora hominis. This Vavraia-like microsporidium was present in dead salamanders at SMARC archived between 2011 and 2015 (positive rates ranging between 52.0–88.9% by PCR detection), as well as in some aquatic invertebrates at the facility (e.g. snails and small crustaceans). Conclusions A Vavraia-like microsporidian was at least one of the major pathogens, if not solely, responsible for the sickness and mortality in the SMARC salamanders, and the pathogen had been present in the center for years. Environmental invertebrates likely served as a source and reservoir of the microsporidian pathogen. These observations provide new knowledge and a foundation for future conservation efforts for Eurycea salamanders including molecular surveys, monitoring of the pathogen, and discovery of effective treatments.

Microsporidian infections of insects are important natural constraints of population growth, often reducing lifespan, fecundity and fertility of the infected host. The recently discovered Tubulinosema suzukii infects Drosophila suzukii (spotted wing drosophila, SWD), an invasive pest of many fruit crops in North America and Europe. In laboratory tests, fitness effects on larval and adult stages were explored. High level infection after larval treatment caused up to 70% pupal mortality, a decreased lifespan and a 70% reduced oviposition of emerging adults in biparental infection clusters. A shift to higher proportion of female offspring compared to controls suggested a potential parthenogenetic effect after microsporidian infection. A clear sex-linkage of effects was noted; females were specifically impaired, as concluded from fecundity tests with only infected female parents. Additive effects were noted when both parental sexes were infected, whereas least effects were found with only infected male parents, though survival of males was most negatively affected if they were fed with T. suzukii spores in the adult stage. Although most negative effects on fitness parameters were revealed after larval treatment, infection of offspring was never higher than 4%, suggesting limited vertical transmission. For that reason, a self-reliant spread in natural SWD populations would probably only occur by spore release from cadavers or frass. [ABSTRACT FROM AUTHOR]

Introduction: Human microsporidiosis represents an important and rapidly emerging opportunistic disease. The present study investigated the prevalence of microsporidia among HIV positive and HIV negative patients with or without diarrhoea in Vhembe and Mopani Districts in the Limpopo Province. Methodology: A total of 170 stool samples were collected from these patients and microsporidia species was detected using a Real-Time PCR targeting a conserved region of the small ribosomal subunit rRNA (SSU-rRNA) gene of Enterocytozoon bieneusi, Encephalitozoon intestinalis, Encephalitozoon hellem, and Encephalitozoon cuniculi. Results: Fifty six (32.9%) were positive for microsporidia. The prevalence was higher in HIV negative patients (36.6%) while 24.1% of patients who were HIV positive had microsporidia. Microsporidia was more common among patients aged between 1 and 10 years (52.6%). However among the HIV positive patients, microsporidia prevalence was higher among those that were not taking antiretrovirals (ARVs) compared to those who were on ARVs, (36.6%) and (24.1%), respectively. Microsporidia was also noted to be significantly associated with diarrheal and stomach pains; p = 0.02 and p = 0.048, respectively. Furthermore, microsporidia infections was more prevalent among patients who had animals at home (p = 0.037). Conclusions: Study has shown a high prevalence of microsporidia among patients attending primary health centers in the Mopani District for the first time. Prevalence of microsporidia was higher among HIV negative and HIV positive patients who were not on ARV treatment. Keeping animals in the household appeared to be a risk of getting infected with microsporidia. Further studies are needed to determine the genetic characteristics of these organisms in the study population. [ABSTRACT FROM AUTHOR]

Microsporidiosis is an emerging opportunistic infection causing severe digestive disorders in immunocompromised patients. The aim of this study was to investigate the prevalence of intestinal microsporidia carriage among immunocompromised patients hospitalized at a major hospital complex in the Tunis capital area, Tunisia (North Africa), and perform molecular epidemiology and population structure analyses of Enterocytozoon bieneusi, which is an emerging fungal pathogen. We screened 250 stool samples for the presence of intestinal microsporidia from 171 patients, including 81 organ transplant recipients, 73 Human Immunodeficiency Virus (HIV)-positive patients, and 17 patients with unspecified immunodeficiency. Using a nested PCR-based diagnostic approach for the detection of E. bieneusi and Encephalitozoon spp., we identified 18 microsporidia-positive patients out of 171 (10.5%), among which 17 were infected with E. bieneusi. Microsporidia-positive cases displayed chronic diarrhea (17 out of 18), which was associated more with HIV rather than with immunosuppression other than HIV (12 out of 73 versus 6 out of 98, respectively, p = 0.02) and correlated with extended hospital stays compared to microsporidia-negative cases (60 versus 19 days on average, respectively; p = 0.001). Strikingly, internal transcribed spacer (ITS)-based genotyping of E. bieneusi strains revealed high-frequency occurrence of ITS sequences that were identical (n = 10) or similar (with one single polymorphic site, n = 3) to rare genotype WL12. Minimum-spanning tree analyses segregated the 17 E. bieneusi infection cases into four distinct genotypic clusters and confirmed the high prevalence of genotype WL12 in our patient population. Phylogenetic analyses allowed the mapping of all 17 E. bieneusi strains to zoonotic group 1 (subgroups 1a and 1b/1c), indicating loose host specificity and raising public health concern. Our study suggests a probable common source of E. bieneusi genotype WL12 transmission and prompts the implementation of a wider epidemiological investigation. View Full-Text [ABSTRACT FROM AUTHOR]

Background: Intestinal microsporidiosis due to Enterocytozoon bieneusi is a cause of chronic diarrhoea in immunocompromised patients. Fumagillin has been approved in France for its treatment.Objectives: To investigate the efficacy and safety of fumagillin in a real-life setting.Methods: As required by the French Medicine Agency, all patients receiving fumagillin were enrolled in a prospective study to evaluate its efficacy and safety. Stool examination with identification of E. bieneusi by PCR was performed at baseline, end of treatment and monthly thereafter for 6 months. Safety was monitored up to 6 months and full blood counts were monitored up to 42 days after treatment initiation. The primary endpoint was safety. Parasite clearance and relapses were secondary endpoints.Results: From 2007 to 2018, 166 patients received fumagillin, including 6 children. Patients were transplant recipients (84%), HIV-infected patients (13%) or had another cause of immunosuppression (5%). Serious adverse events were reported in 41 patients (25%), mainly thrombocytopenia (15%) and neutropenia (5%), with two haemorrhagic events leading to one death. Severe thrombocytopenia (<50 G/L) developed in 50 patients (29.6%), neutropenia (<1 G/L) in 20 patients (11.8%) and severe anaemia (<8 g/dL) in 21 patients (12.4%). At the end of treatment, 94% of patients with available stool examination (n = 132) had no spores detected. Among 99 patients with available follow-up after the end of treatment, three parasite relapses were documented.Conclusions: E. bieneusi microsporidiosis was mainly diagnosed in transplant recipients. Fumagillin was associated with haematological toxicity but showed high efficacy with a low relapse rate. [ABSTRACT FROM AUTHOR]

A novel microsporidial disease was documented in two ornamental fish species, black tetra Gymnocorymbus ternetzi Boulenger 1895 and cardinal tetra Paracheirodon axelrodi Schultz 1956. The non-xenoma-forming microsporidium occurred diffusely in most internal organs and the gill, thus referring to the condition as tetra disseminated microsporidiosis (TDM). The occurrence of TDM in black tetra was associated with chronic mortality in a domestic farmed population, while the case in cardinal tetra occurred in moribund fish while in quarantine at a public aquarium. Histology showed that coelomic visceral organs were frequently necrotic and severely disrupted by extensive infiltrates of macrophages. Infected macrophages were presumed responsible for the dissemination of spores throughout the body. Ultrastructural characteristics of the parasite developmental cycle included uninucleate meronts directly in the host cell cytoplasm. Sporonts were bi-nucleated as a result of karyokinesis and a parasite-produced sporophorous vesicle (SPV) became apparent at this stage. Cytokinesis resulted in two spores forming within each SPV. Spores were uniform in size, measuring about 3.9 ± 0.33 long by 2.0 ± 0.2 μm wide. Ultrastructure demonstrated two spore types, one with 9–12 polar filament coils and a double-layered exospore and a second type with 4–7 polar filament coils and a homogenously electron-dense exospore, with differences perhaps related to parasite transmission mechanisms. The 16S rDNA sequences showed closest identity to the genus Glugea (≈ 92%), though the developmental cycle, specifically being a non-xenoma-forming species and having two spores forming within a SPV, did not fit within the genus. Based on combined phylogenetic and ultrastructural characteristics, a new genus (Fusasporis) is proposed, with F. stethaprioni n. gen. n. sp. as the type species. [ABSTRACT FROM AUTHOR]

The Indian meal moth, Plodia interpunctella (Lepidoptera: Pyralidae) is one of the most important stored product pests. Fumigation plays a significant role in the management of insect pests in stored-products. However, the use of fumigants is problematic because of their effects on the environment and high costs. Entomopathogenic organisms are environmentally friendly control agents and suppress pest populations under natural conditions. In this study, distribution and occurrence of a microsporidian pathogen, Vairimorpha plodiae (Opisthokonta: Microspora) in the populations of P. interpunctella from 12 localities representing Turkey between 2019 and 2020 are presented for the first time by confirming its effectiveness on natural populations. The presence of the microsporidian pathogen was found in 11 of 12 (91.7%) populations. In total, 863 of 3,044 samples were infected by the pathogen. Infection mean was 28.4% for all populations. Our results showed that V. plodiae infection reached to a considerably high prevalence (88.77%) in P. interpunctella populations and varied from 5.1 to 88.7% between the populations. In addition, microsporidia infections have been identified throughout Turkey. We found that V. plodiae can infect all life stages of P. interpunctella. Totally, 623 (28.5%) of 2187 larvae, 14 (37.8%) of 37 pupae, 226 (27%) of 820 adults were found to be infected by the pathogen. There were considerable differences between the dead and living larvae. The microsporidian infection was found in 26 (11.6%) of 225 living larvae, whereas it was found in 595 (30.5%) of 1,952 dead larvae. These results confirm that the microsporidia pathogen has a high spreading potential in P. interpunctella populations and can be a natural biological suppression factor on pest populations. [ABSTRACT FROM AUTHOR]

Microsporidia are a group of unicellular and opportunistic intestinal parasites in which Enterocytozoon bieneusi is a frequent species causing microsporidial infections in humans. Many domesticated and wild animals have been shown to be hosts of E. bieneusi and other microsporidia. The role of hedgehogs in the ecology of microsporidia is unclear; therefore, we investigated the prevalence and genetic diversity of E. bieneusi, Cryptosporidium, and Blastocystis spp. in hedgehogs (Erinaceus amurensis) collected from Hubei Province in Central China. PCR amplification of the internal transcribed spacer region of the ribosomal DNA indicated that 9.8% (4/41) hedgehogs were positive to E. bieneusi, but none (0/41) was positive to Cryptosporidium and Blastocystis spp. Phylogenetic analysis showed the strains detected from the hedgehogs belong to four novel genotypes (EA1–EA4), which were most closely related to type IV of group 1c. This study demonstrated that hedgehogs are hosts of E. bieneusi and may play a role in the transmission of E. bieneusi to humans in the process of being caught and slaughtered. [ABSTRACT FROM AUTHOR]

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 Córdoba, 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. [ABSTRACT FROM AUTHOR]

Encephalitozoon cuniculi genotype III disseminated intensively into most of the organs in all strains of mice, followed by a chronic infection with massive microsporidia persistence in immunodeficient mice and a partial decrease in C57Bl/6 mice. Treatment with 0.2 mg Albendazole/mouse/day temporarily reduces the number of affected organs in immunocompetent C57Bl/6 mice, but not in CD4−/− and CD8−/− mice. The application of medication temporarily decreased the spore burden at least by one order of magnitude in all groups. These results demonstrate that the E. cuniculi genotype III infection had a progressive course and surprisingly, Albendazole treatment had only a minimal effect. The E. cuniculi genotype III spore burden in individual organs reached up to 108 or 109 in immunocompetent or immunodeficient mice, respectively; however, these mice did not demonstrate any obvious clinical signs of microsporidiosis, and the immunodeficient mice survived longer. Our findings clearly show that the survival of mice does not correspond to spore burden, which provides new insight into latent microsporidiosis from an epidemiological point of view. [ABSTRACT FROM AUTHOR]

• NO level and NOS activity were influenced by microsporidia immune stimulation. • AMP and immune genes can be influenced by NO content in Antheraea pernyi gut. • Microsporidia infection rate change under NOS inhibitor treatment. The insect gut participates in initial local immune responses by producing reactive oxygen and nitrogen species as well as anti-microbial peptides to resist pathogenic invasions. Nitric oxide (NO), a signaling and an immune effector molecule synthesized by the enzyme NO synthase (NOS), mediates an early step of the signal transduction pathway. In this study, we evaluated NO levels after Nosema pernyi infection in Antheraea pernyi gut. NOS activity was higher in the microsporidia-infected gut of A. pernyi than in that of control. Three NOS -related genes were cloned, and their spatio-temporal expression patterns were evaluated. ApNOS2 was expressed quickly in the midgut after N. pernyi infection. Sodium nitroprusside, dihydrate (SNP), or Nω- L -nitro-arginine methyl ester, hydrochloride (L -NAME), altered the NO content in A. pernyi midgut. Anti-microbial peptides (AMPs) in the groups exposed to N. pernyi plus SNP and N. pernyi plus L -NAME exhibited higher and lower expression, respectively, relative to the control. These results indicate that microsporidia infection triggers short-term activation of NO and NOS genes in the A. pernyi gut that is downregulated after 24 h. Notably, infection rates can be influenced by a NOS inhibitor. Furthermore, NO can be induced by pathogens. Similarly, NO content in the A. pernyi gut also influences AMPs in humoral immunity and some immune-related genes. Our results suggest that nitric oxide plays a vital role in A. pernyi gut immunity. [ABSTRACT FROM AUTHOR]

Microsporidia, a divergent group of single-celled eukaryotic parasites, harness a specialized harpoon-like invasion apparatus called the polar tube (PT) to gain entry into host cells. The PT is tightly coiled within the transmissible extracellular spore, and is about 20 times the length of the spore. Once triggered, the PT is rapidly ejected and is thought to penetrate the host cell, acting as a conduit for the transfer of infectious cargo into the host. The organization of this specialized infection apparatus in the spore, how it is deployed, and how the nucleus and other large cargo are transported through the narrow PT are not well understood. Here we use serial block-face scanning electron microscopy to reveal the 3-dimensional architecture of the PT and its relative spatial orientation to other organelles within the spore. Using high-speed optical microscopy, we also capture and quantify the entire PT germination process of three human-infecting microsporidia species in vitro: Anncaliia algerae, Encephalitozoon hellem and E. intestinalis. Our results show that the emerging PT experiences very high accelerating forces to reach velocities exceeding 300 μm⋅s-1, and that firing kinetics differ markedly between species. Live-cell imaging reveals that the nucleus, which is at least 7 times larger than the diameter of the PT, undergoes extreme deformation to fit through the narrow tube, and moves at speeds comparable to PT extension. Our study sheds new light on the 3-dimensional organization, dynamics, and mechanism of PT extrusion, and shows how infectious cargo moves through the tube to initiate infection. Author summary: Microsporidia infect a wide range of hosts: from economically important invertebrates such as silkworms and honey bees, to vertebrates including humans, where infection in immunocompromised patients can be fatal. In order to infect the host, microsporidia utilize a unique harpoon-like invasion organelle called the polar tube (PT), which serves as a pathway for transport of infectious cargo to the host. Knowledge of how the long PT is packaged in the spore, the kinetics of its firing and how the cargo is transported through the PT are still poorly understood. We use electron microscopy techniques to generate 3D reconstructions of intact spores, which reveal the configuration of the PT and other organelles within. Using high-speed live-cell imaging, we capture and quantify the kinetics of PT firing, and how cargo is deformed during transport through the narrow PT. Our study provides new insights into architecture and dynamics of the PT, which serve as foundations for our understanding of microsporidia infection. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Microsporidia Amblyospora opacita Kudo, 1922 was tested in laboratory conditions as an agent of biological control of vectors of pathogens of many dangerous infectious and invasive diseases - blood-sucking mosquitoes (Aedes caspius Pallas, 1771). The effect of microsporidiosis on mortality, growth, developmental period, molting and sex of larvae was studied. It was found that the prepared suspension titer of 1.1×105 spores / ml is more effective (up to 90.7%) against the larvae of bloodsucking mosquitoes. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

• Success of kidney and/or pancreas transplantation is sometimes tarnished by opportunistic infections such as cryptosporidiosis and microsporidiosis, two neglected infections in developed countries. • We conducted the first prospective study in a French referral center for kidney-pancreas transplantation over a one-year period to gain further insights into the prevalence and risk factors for cryptosporidiosis and microsporidiosis. All patients declaring to have diarrhea underwent, at inclusion, a systematic and standardized etiological exploration and an evaluation of their immunity. A questionnaire about environmental risk factors was also administered. Our prospective study provides new insights and is the first to underline both cryptosporidiosis and microsporidiosis as significant causes of diarrhea in kidney transplant recipients. Gastrointestinal disorders in solid organ recipients may have various origins including cryptosporidiosis and microsporidiosis. The prevalence of these infections is poorly known in solid organ transplant (SOT) patients in industrialized countries. We prospectively assessed the infectious causes of diarrhea in SOT patients. Secondary objectives were to gain further insight into the main characteristics of cryptosporidiosis, and to assess risk factors for this infection. All adult kidney and/or pancreas recipients presenting with diarrhea and admitted to our facility between May 1, 2014 and June 30, 2015 were enrolled. A stool sample was analyzed using a standardized protocol including bacteriological, virological, and parasitological investigations. Data related to clinical symptoms, immunosuppression, and environmental potential risk factors were collected through a self-administered questionnaire and computerized medical records. Out of 73 enrolled patients, 36 had infectious diarrhea (49.3%). Viruses ranked first (17/36), followed by parasites and fungi (11/17). Cryptosporidiosis was the most common parasitic disease (n = 6 patients). We observed four microsporidiosis cases. The estimated prevalence of cryptosporidiosis and microsporidiosis in this cohort was 3.7 and 2.40/ 00 , respectively. No significant risk factor for cryptosporidiosis or microsporidiosis, neither environmental nor immunological, could be evidenced. Both cryptosporidiosis and microsporidiosis represent a significant cause of diarrhea in kidney transplant recipients. [ABSTRACT FROM AUTHOR]

Nosema ceranae is a microsporidian parasite that is pathogenic to honey bees, causing disease at the individual and colony level. N. ceranae infection can be controlled by treatment with Fumagillin, but as this the future of this drug is uncertain, alternative treatment strategies are critical. Genomic examination of proteostasis pathways in N. ceranae and its host, Apis mellifera, has revealed key differences in component conservation. One example is the amino acid response (AAR), which responds to amino acid limitation through the sensing of uncharged tRNA molecules. The AAR is conserved in the honey bee, but not in N. ceranae. Pharmacological inhibition of prolyl-tRNA synthetase activity resulted in a substantial reduction in N. ceranae infection intensity. Evident toxicity to bees suggests further work is required to make this approach safe and feasible. However, these results provide proof of principle that tRNA synthetase inhibition could be an effective future treatment strategy. [ABSTRACT FROM AUTHOR]

Nosema ceranae infections in honey bees (Apis mellifera) pose a severe threat to colony health. Beekeepers have used dicyclohexylammonium fumagillin to control Nosema apis , although it may be ineffective against N. ceranae. We investigated the ability of various propolis extracts collected from Upstate New York (United States) to decrease in vivo N. ceranae infection levels when fed ad libitum to N. ceranae -infected honey bees. Propolis extracts, most notably a dichloromethane extract, significantly lowered spore levels in a dose-dependent fashion 4 days post inoculation. When testing the in vitro anti- Nosema activity of propolis extracts, we report for the first time that spore viability was unaffected after a 24 h exposure to propolis extracts. These results present evidence that propolis extracts may effectively lower Microsporidia infections in honey bees, and that direct exposure of environmental spores to propolis alone does not kill N. ceranae. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Enterocytozoon bieneusi is a human pathogen with a broad range of animal hosts. Initially, E. bieneusi was considered an emerging opportunistic pathogen in immunocompromised, mainly HIV-infected patients, but it has been increasingly reported in apparently healthy individuals globally. As in other African countries, the molecular epidemiology of E. bieneusi in Mozambique remains completely unknown. Therefore, we undertook a study to investigate the occurrence and genetic diversity of E. bieneusi infections in children with gastrointestinal symptoms as well as in asymptomatic children in Mozambique. Individual stool specimens were collected from 1,247 children aged between 0 and 14 years-old living in urban and rural settings in Zambézia (n = 1,097) and Maputo (n = 150) provinces between 2016 and 2019. Samples were analysed for E. bieneusi by nested-PCR targeting the internal transcribed spacer (ITS) region of the rRNA gene. All positive amplicons were confirmed and genotyped. Penalised logistic regression (Firth) was used to evaluate risk associations. The overall prevalence of E. bieneusi in this children population was 0.7% (9/1,247). A 10-fold higher prevalence was found in Maputo (4.0%; 6/150) than in Zambézia (0.3%; 3/1,097). All E. bieneusi-positive samples were from children older than 1-year of age, and most (8/9) from asymptomatic children. Nucleotide sequence analysis of the ITS region revealed the presence of four genotypes, three previously reported (Peru11, n = 1; Type IV, n = 2, and S2, n = 2) and a novel genotype (named HhMzEb1, n = 4). Novel genotype HhMzEb1 was identified in both asymptomatic (75%, 3/4) and symptomatic (25%, 1/4) children from a rural area in Maputo province in southern Mozambique. Genotypes HhMzEb1, Peru11, S2, and Type IV belonged to the Group 1 that includes genotypes with low host specificity and the potential for zoonotic and cross-species transmission. Being infected by enteric protozoan parasites and no handwashing were identified as risk associations for E. bieneusi infection. This study reports the first investigation of E. bieneusi genotypes in Mozambique with the identification of three previously reported genotypes in humans as well as a novel genotype (HhMzEb1). Findings highlight the need to conduct additional research to elucidate the epidemiology of E. bieneusi in the country, especially in rural areas where poor hygiene conditions still prevail. Special attention should be paid to the identification of suitable animal and environmental reservoirs of this parasite and to the characterization of transmission pathways. Author summary: Enterocytozoon bieneusi is an obligate intracellular parasite that infects a wide range of vertebrate hosts. It is the most important etiological agent of human microsporidiasis. Most clinical and epidemiological studies conducted to date have focused on immunodeficient or immunosuppressed individuals including HIV+ patients and solid organ transplant recipients, as in those E. bieneusi infection causes life-threatening chronic diarrhoea. In contrast, latent microsporidia infections in immunocompetent individuals have received far less attention. Molecular epidemiological studies in humans and animals have revealed that E. bieneusi encompasses a very large diversity of genetic variants (genotypes) with marked differences in host specificity and even geographical distribution. In Mozambique, as in many other African countries, the epidemiology of E. bieneusi is completely unknown. Therefore, to identify the occurrence and genetic diversity of this pathogen in Mozambique stool samples were obtained from children, including apparently healthy and symptomatic, in Zambézia and Maputo provinces and tested for E. bieneusi by molecular methods. Results demonstrated the presence of E. bieneusi genotypes for the first time in Mozambique. Four genotypes were identified, three genotypes that have previously been reported in humans (Peru11, Type IV, and S2) and a novel genotype (HhMzEb1). Two of the genotypes Peru11, Type IV have also been frequently identified in animals indicating that potentially zoonotic E. bieneusi genotypes are inadvertently circulating in the surveyed populations. Additional population genetic studies are needed to elucidate the actual extent of the epidemiology and transmission dynamics of E. bieneusi in Mozambique. [ABSTRACT FROM AUTHOR]

Enterocytozoon bieneusi is one of the most common intestinal pathogens in humans and animals. E. bieneusi has been confirmed to be complex microsporidian species. Approximately 500 ITS genotypes of E. bieneusi have been defined. With the establishment and application of multilocus sequencing typing and population genetic tools in E. bieneusi , the studies on these aspects have been carried out worldwide, but little information is available. To understand genetic variation of mini-/micro-satellites and the population structure and substructure of E. bieneusi in northeastern China, 305 E. bieneusi DNA specimens composed of 28 ITS genotypes were from 13 mammal species and five bird species in the investigated areas. They were characterized by nested-PCR amplification and sequencing at four mini-/micro-satellite loci (MS1, MS3, MS4, and MS7). At the MS1, MS3, MS4, and MS7 loci, 153 (50.16%), 131 (42.95%), 133 (43.61%), and 128 (41.97%) DNA specimens were amplified and sequenced successfully with 44, 17, 26, and 24 genotypes being identified, respectively. Multilocus genotypes (MLGs) showed a higher genetic diversity than ITS genotypes. 48 MLGs were produced out of 90 ITS-positive DNA specimens based on concatenated sequences of all the five genetic loci including ITS. Linkage disequilibrium (LD) and limited genetic recombination were observed by measuring LD using both multilocus sequences and allelic profile data, indicating an overall clonal population structure of E. bieneusi in the investigated areas. These data will aid in the longitudinal tracking of the attribution of source of infection/contamination and in elucidating transmission dynamics, and will provide valuable information for making efficient control strategies to intervene with and prevent occurrence of microsporidiosis caused by E. bieneusi among animals and transmission of E. bieneusi from animals to humans in the investigated areas. Phylogenetic and network analyses identified three different subgroups, revealing the presence of host-shaped segregation and the absence of geographical segregation in E. bieneusi population. Meanwhile, the MLGs from zoonotic ITS genotypes were observed to be basically separated from the MLGs from host-adapted ones. Assessment of substructure will have a reference effect on understanding of zoonotic or interspecies transmission of E. bieneusi and evolution direction from zoonotic genotypes to host-adapted genotypes. [ABSTRACT FROM AUTHOR]

Microsporidia are obligate intracellular and eukaryotic pathogens that can infect immunocompromised and immunocompetent mammals, including humans. Both innate and adaptive immune systems play important roles against microsporidian infection. The innate immune system can partially eliminate the infection by immune cells, such as gamma delta T cell, natural killer cells (NKs), macrophages and dendritic cells (DCs), and present the pathogens to lymphocytes. The innate immune cells can also prime and enhance the adaptive immune response via surface molecules and secreted cytokines. The adaptive immune system is critical to eliminate microsporidian infection by activating cytotoxic T lymphocyte (CTL) and humoral immune responses, and feedback regulation of the innate immune mechanism. In this review, we will discuss the cellular and molecular responses and functions of innate and adaptive immune systems against microsporidian infection. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Enterocytozoon bieneusi is the mainly pathologies or intestinal disorders that causes approximately 90% of reported cases of human microsporidiosis. To understand the prevalence and genotype distribution of E. bieneusi in the Xinjiang Uygur Autonomous Region, China, 609 fecal samples were collected from children in kindergarten in Southern Xinjiang and screened for this pathogen by PCR and sequencing of the internal transcribed spacer (ITS). Thirty-six fecal samples (5.9%, 36/609) were positive for E. bieneusi, with the highest prevalence observed in children from Yopurga (17.5%, 11/63). Nine genotypes were identified, of which six were known (A, CHN6, D, EbpA, KB-1, and NIA1) and three were novel (CXJH1, CXJH2 and CXJH3). Genotype NIA1 was most prevalent (52.8%, 19/36), followed by genotypes D (16.7%, 6/36), A (8.3%, 3/36), and EbpA (8.3%, 3/36). The remaining five genotypes were detected in one sample each. Phylogenetic analysis revealed that the E. bieneusi isolates clustered into two groups, one consisting of six genotypes (Group 1: A, CXJH1, D, EbpA, KB-1, and NIA1) and another consisting of three genotypes (Group 2: CHN6, CXJH2, and CXJH3). Our results confirmed that infection of E. bieneusi unusual dominant genotype NIA1 occurs in children in Xinjiang, China. Further epidemiological studies must be conducted to clarify potential sources of E. bieneusi infection in this area. Author summary: This study reports the infection rates and genetic characteristics of Enterocytozoon bieneusi in 609 children in kindergarten in Southern Xinjiang, China. All samples were screened for this pathogen with PCR, based on the internal transcribed spacer (ITS) of E. bieneusi. Thirty-six fecal samples (5.9%, 36/609) were positive, with the highest prevalence observed in children from Yopurga (17.5%, 11/63). Three novel genotypes were identified (CXJH1, CXJH2 and CXJH3). Phylogenetic analysis revealed that the E. bieneusi isolates clustered into two groups: Group 1 (A, CXJH1, D, EbpA, KB-1, and NIA1) and Group 2 (CHN6, CXJH2, and CXJH3). Genotype NIA1 used to be detected in HIV-positive patients, however, it was most prevalent (52.8%, 19/36) among the nine genotypes identified in this study. Additionally, we confirmed the zoonotic potential of E. bieneusi genotype D and this is the first report of human infection by E. bieneusi genotypes KB-1 and CHN6. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Sulbutiamine is a thiamine derivative developed in Japan in the mid-60's as a beriberi treatment drug. Since then, different potential applications have been described. For instance, there is some evidence that sulbutiamine can have anti-fatigue, nootropic, and antioxidant effects, which led to its use as a sport supplement (although some authors argue it is actually a masking doping strategy). Moreover, this molecule has been proposed as a possible treatment for some microsporidial infections and even for certain types of cancer. Despite these potential effects, sulbutiamine is still a relatively unknown molecule, which justifies the present review, where we discuss its history and the existing literature on its health applications. We conclude that there is a great potential for sulbutiamine use, well beyond its first described function (to increase thiamine tissue concentration). Indeed, new mechanisms of action have been found, mainly associated with its derivatives. Nevertheless, and although the research on sulbutiamine started 50 years ago, only a limited number of studies were conducted during this time frame. As so, methodological concerns need to be addressed and new studies are necessary, especially randomized controlled trials. Only then will the full potential of this versatile molecule be identified. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Microsporidia are found worldwide and both vertebrates and invertebrates can serve as hosts for these organisms. While microsporidiosis in humans can occur in both immune competent and immune compromised hosts, it has most often been seen in the immune suppressed population, e.g., patients with advanced HIV infection, patients who have had organ transplantation, those undergoing chemotherapy, or patients using other immune suppressive agents. Infection can be associated with either focal infection in a specific organ (e.g., keratoconjunctivitis, cerebritis, or hepatitis) or with disseminated disease. The most common presentation of microsporidiosis being gastrointestinal infection with chronic diarrhea and wasting syndrome. In the setting of advanced HIV infection or other cases of profound immune deficiency microsporidiosis can be extremely debilitating and carries a significant mortality risk. Microsporidia are transmitted as spores which invade host cells by a specialized invasion apparatus the polar tube (PT). This review summarizes recent studies that have provided information on the composition of the spore wall and PT, as well as insights into the mechanism of invasion and interaction of the PT and spore wall with host cells during infection. [ABSTRACT FROM AUTHOR]

Microsporidiosis is a rare, but emerging opportunistic infection in solid organ transplant and stem cell transplant recipients. Renal involvement in microsporidiosis is very rarely seen in these recipients. We describe two cases of pediatric renal microsporidiosis, diagnosed on renal biopsies, following bone marrow transplantation presenting as severe acute kidney injury. The first patient died, whereas the second survived due to early diagnosis based on high index of suspicion and prompt treatment with Albendazole. We believe these are the first such reported cases of renal microsporidiosis in pediatric bone marrow transplant recipients. [ABSTRACT FROM AUTHOR]

Representatives of the genus Anncaliia are known as natural parasites of dipteran and coleopteran insects, amphipod crustaceans, but also humans, primarily with immunodeficiency. Anncaliia algerae‐caused fatal myositis is considered as an emergent infectious disease in humans. A. (=Nosema, Brachiola) algerae, the best studied species of the genus, demonstrates the broadest among microsporidia range of natural and experimental hosts, but it has never been propagated in Drosophila. We present ultrastructural analysis of development of A. algerae in visceral muscles and adipocytes of Drosophila melanogaster 2 wk after per oral experimental infection. We observed typical to Anncaliia spp. features of ultrastructure and cell pathology including spore morphology, characteristic extensions of the plasma membrane, and presence of "ridges" and appendages of tubular material at proliferative stages. Anncaliia algerae development in D. melanogaster was particularly similar to one of A. algerae and A.(Brachiola) vesicularum in humans with acute myositis. Given D. melanogaster is currently the most established genetic model, with a fully sequenced genome and easily available transgenic forms and genomic markers, a novel host–parasite system might provide new genetic tools to investigate host–pathogen interactions of A. algerae, as well to test antimicrosporidia drugs. [ABSTRACT FROM AUTHOR]

The microsporidian Nosema bombycis is an obligate intracellular parasitic fungus that causes devastating disease in sericulture. To date, no efficient biotechnological method to inhibit the proliferation of microspores has been established. Here, we developed a powerful genetic engineering technique involving microsporidia-inducible genome editing in transgenic silkworm that confers resistance to N. bombycis. This system includes an HSP70 promoter-induced expression of the Cas9 protein line and a target BmATAD3A gene line. The double-positive HSP70-Cas9(+)×sgATAD3A(+) lines were obtained by hybridization and activation of the CRISPR/Cas9 system under the condition of microsporidia infection, although it is silenced in uninfected individuals. Genome editing analysis showed that the system could efficiently edit the BmATAD3A gene and induce large deletions. It is notable that the HSP70-induced system could effectively improve the survival rate of transgenic silkworm after microsporidia infection and inhibit the expression of key microsporidia genes. Moreover, no significant developmental differences between the transgenic silkworms infected with microsporidia and normal individuals were observed. In this study, we effectively inhibited microsporidia proliferation in transgenic individuals through disruptive techniques, thereby providing a method for microsporidia treatment and prevention, paving the way for economically advantageous insect breeding. [ABSTRACT FROM AUTHOR]

Enterocytozoon hepatopenaei (EHP) is abundant in the shrimp hepatopancreas and intestines and can be transmitted through the digestive system. But the relationship between EHP and intestines or gut microbiota is still unclear. In this study, the microbiome and proteome of shrimp Penaeus vannamei intestine were used to analyze the changes of the gut microbiome and the immune protein after EHP infection. Our results showed that EHP infection brought about a marked increase in the proportion of pathogenic bacteria, such as Shigella , Aeromonas , Faecalibacterium , and Streptococcus , suggesting that EHP infection would lead to the increasing of pathogenic bacteria. Proteome results indicated showed that down-regulated proteins were mainly concentrated in immune-related domains such as C-type lectins (CTLs), hemoglobin, and glutathione transferase, suggesting that EHP might lead to a weakening of the innate immunity of shrimp. In shrimp, CTLs can involve in innate immunity by regulating the transcription of various antimicrobial peptides, and directly participate in the inhibition of pathogens through binding and agglutination. On this basis, after successfully cloned and recombination expressed, three down-regulated CTLs (perlucin, mannose receptor 1 and c-type lectin domain family 17 with typical CTL domains) were proved to have binding and agglutinin ability against four pathogenic bacteria (Enterococcus faecalis , Shigella castellani , Salmonella enteritidis , Aeromonas hydrophila). These results suggested that EHP infection led to decrease the expression of three CTLs and increase secondary infection caused by pathogenic bacteria. And CTLs played important roles in maintaining the balance of shrimp gut microbiome in microsporidia infection. In a word, these results are helpful for us to better understand the interaction between EHP and intestinal microbiota of P. vannamei and will gain new insights into prevention and control of hepatopancreatic microsporidiosis. [Display omitted] • Enterocytozoon hepatopenaei infection disrupted intestinal microbiota and reduced the immunity of P. vannamei. • Three CTLs (PvPer, PvMR1 and PvCTL17) could bind and agglutinate directly to all tested bacterial pathogens • PvPer, PvMR1 and PvCTL17 might regulate intestinal microbiome homeostasis via inhibiting pathogenic bacteria. [ABSTRACT FROM AUTHOR]

Ameson portunus (Microsporidia) has caused serious economic losses to the aquaculture industry of swimming crab, Portunus trituberculatus. The hemolymph and hepatopancreas are the main immune organs of P. trituberculatus , and the main sites of A. portunus infection. Elucidating the response characteristics of hemolymph and hepatopancreas to microsporidian infection facilitates the development of microsporidiosis prevention and control strategy. This study performed comparative transcriptomic analysis of hemolymph (PTX/PTXA) and hepatopancreas (PTG/PTGA) of P. trituberculatus uninfected and infected with A. portunus. The results showed that there were 223 and 1309 differentially expressed genes (DEGs) in PTX/PTXA and PTG/PTGA, respectively. The lysosome pathway was significantly enriched after the invasion of the hemolymph by A. portunus. Also, immune-related genes were all significantly up-regulated in the hemolymph and hepatopancreas, suggesting that the invasion by A. portunus may activate host immune responses. Unlike hemolymph, antioxidant and detoxification-related genes were also significantly up-regulated in the hepatopancreas. Moreover, metabolism-related genes were significantly down-regulated in the hepatopancreas, suggesting that energy synthesis, resistance to pathogens, and regulation of oxidative stress were suppressed in the hepatopancreas. Hemolymph and hepatopancreas have similarity and tissue specificity to microsporidian infection. The differential genes and pathways identified in this study can provide references for the prevention and control of microsporidiosis. • Hemolymph and hepatopancreas transcriptomes of Portunus trituberculatus infected with Ameson portunus were analyzed. • Upregulation of immune-related genes suggests that microsporidian infection activates host immune responses. • Downregulation of metabolism-related genes affects host energy synthesis, resistance to pathogens, oxidative stress regulation. • Hemolymph and hepatopancreas have similarity and tissue specificity to microsporidian infection. [ABSTRACT FROM AUTHOR]

In April 2014, a kidney transplant recipient in the United States experienced headache, diplopia, and confusion, followed by neurologic decline and death. An investigation to evaluate the possibility of donor-derived infection determined that 3 patients had received 4 organs (kidney, liver, heart/kidney) from the same donor. The liver recipient experienced tremor and gait instability; the heart/kidney and contralateral kidney recipients were hospitalized with encephalitis. None experienced gastrointestinal symptoms. Encephalitozoon cuniculi was detected by tissue PCR in the central nervous system of the deceased kidney recipient and in renal allograft tissue from both kidney recipients. Urine PCR was positive for E. cuniculi in the 2 surviving recipients. Donor serum was positive for E. cuniculi antibodies. E. cuniculi was transmitted to 3 recipients from 1 donor. This rare presentation of disseminated disease resulted in diagnostic delays. Clinicians should consider donor-derived microsporidial infection in organ recipients with unexplained encephalitis, even when gastrointestinal manifestations are absent.

Objetivo: Reportar los casos de queratitis estromal por microsporidios de nuestra institución, así como realizar una revisión de la literatura para actualizar el conocimiento sobre los tratamientos y métodos diagnósticos disponibles. Métodos: Se recopilaron un total de 4 casos de queratitis estromal por microsporidios con datos histológicos completos. Se realizó una revisión exhaustiva de la literatura sobre el tema. Resultados: El examen microscópico de los 4 casos reveló con hematoxilina y eosina cúmulos de numerosos organismos redondos a ovales en el estroma corneal, intensamente positivos con las tinciones de Ziehl-Neelsen y azul de toluidina. Conclusión: Esta entidad continúa representando un problema oftalmológico serio debido a su curso indolente y a la mala respuesta a los tratamientos existentes, aunado a que aún no se cuenta con un estándar diagnóstico ampliamente aceptado que permita un diagnóstico certero sin realizar procedimientos invasivos.

Microsporidia are intracellular fungus-like pathogens and the infection symptoms include recurrent diarrhea and systematic inflammations. The major infection route of microsporidia is the digestive tract. Since microsporidia are hard to fully eliminate, the interactions and persistence of the pathogen within epithelium may modulate host susceptibility to digestive disorders. In this study, both in vitro and in vivo infection models were applied. The alterations of epithelial barrier integrity, permeability, and tight junction proteins after microsporidia infection were assessed on MDCK/Caco-2 monolayers. The fecal intestinal microbiota and tissue alterations after microsporidia infection were assessed on C57BL/6 mice. Moreover, the susceptibility to develop dextran sulfate sodium (DSS)-induced inflammatory bowel diseases (IBDs) was also analyzed by the murine infection model. The results demonstrated that microsporidia infection increased epithelium permeability, weakened wound healing capability, and destructed tight junction protein zonula occludens-1. Microsporidia infection also dysregulates intestinal microbiota. These impairing effects of microsporidia increased host vulnerability to develop enteritis as shown by the murine model of DSS-induced IBD. Our study is the first to elucidate molecular mechanisms of the damaging effects of microsporidia on host epithelium and pointed out the cryptic threats of latent microsporidia infection to public health as reflected by the increased chances of developing more severe diseases.IMPORTANCEMicrosporidia are widely present in nature and usually cause latent and persistent infections in hosts. Given the fact that the digestive tract is the major infection route, it is of great importance to explore the consequences of microsporidia infection on the intestinal epithelial barrier and the risks to the host. In this study, we demonstrated the destructing effects of microsporidium infection on epithelial barriers manifested as increased epithelial permeability, weakened wound healing ability, and disrupted tight junctions. Moreover, microsporidia made the host more susceptible to dextran sulfate sodium-induced inflammatory bowel disease. These findings provide new evidence for us to better understand and develop novel strategies for microsporidia prevention and disease control., Competing Interests: The authors declare no conflict of interest.

Importance: The multiplex-crRNA CRISPR/Cas12a detection method saves hands-on time, reduces the risk of aerosol pollution, and can be directly applied to detecting silkworms infected with Nosema bombycis . This study provides a new approach for the inspection and quarantine of silkworm pébrine disease in sericulture and provides a new method for the detection of other pathogens., Competing Interests: The authors declare no conflict of interest.