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Background: Microsporidia MB (MB) is a naturally occurring symbiont of Anopheles and has recently been identified as having a potential to inhibit the transmission of Plasmodium in mosquitoes. MB intensity is high in mosquito gonads, with no fitness consequences for the mosquito, and is linked to horizontal (sexual) and vertical (transovarial) transmission from one mosquito to another. Maximising MB intensity and transmission is important for maintaining heavily infected mosquito colonies for experiments and ultimately for mosquito releases. We have investigated how diet affects the MB-Anopheles arabiensis symbiosis phenotypes, such as larval development and mortality, adult size and survival, as well as MB intensity in both larvae and adults. Methods: F1 larvae of G0 females confirmed to be An. arabiensis and infected with MB were either combined (group lines [GLs]) or reared separately (isofemale lines [IMLs]) depending on the specific experiment. Four diet regimes (all mg/larva/day) were tested on F1 GLs: Tetramin 0.07, Tetramin 0.3, Gocat 0.3 and Cerelac 0.3. GLs reared on Tetramin 0.3 mg/larva/day were then fed either a 1% or 6% glucose diet to determine adult survival. Larvae of IMLs were fed Tetramin 0.07 mg and Tetramin 0.3 mg for larval experiments. The mosquitoes in the adult experiments with IMLs were reared on 1% or 6% glucose. Results: Amongst the four larval diet regimes tested on An.arabiensis development in the presence of MB, the fastest larval development highest adult emergence, largest body size of mosquitoes, highest prevalence and highest density of MB occurred in those fed Tetramin 0.3 mg/larva/day. Although adult MB-positive mosquitoes fed on 6% glucose survived longer than MB-negative mosquitoes, there was no such effect for those fed on the 1% glucose diet. Development time, wing length and adult survival were not significantly different between MB-infected and uninfected An. arabiensis fed on the Tetramin 0.07 mg/larva/day diet, demonstrating that the MB-conferred fitness advantage was diet-dependent. Conclusions: Microsporidia MB does not adversely impact the development and fitness of An. arabiensis, even under limited dietary conditions. The diet regime of Tetramin 0.3 mg/larva/day + 6% glucose for adults is the superior diet for the mass rearing of MB-infected An. arabiensis mosquitoes. These results are important for rearing MB-infected An. arabiensis in the laboratory for experiments and the mass rearing required for field releases. [ABSTRACT FROM AUTHOR]

The inland bearded dragon (Pogona vitticeps) is a lizard species commonly kept as a pet worldwide. Endoparasites are among the most important pathogens affecting bearded dragons. The aim of this study was to evaluate the endoparasites in captive P. vitticeps in Italy. Faecal samples from 30 P. vitticeps were analysed by fresh faecal smears, flotation tests, the Mini-FLOTAC technique, and a rapid immunoassay to detect Cryptosporidium spp. To search for microsporidia, PCR and sequencing were performed on the faecal samples. Data were statistically analysed. The overall positivity rate for endoparasites was 83.3% (25/30). The identified endoparasites were oxyurids (17/30, 56.7%), Isosospora amphiboluri (13/30, 43.3%), Encephalitozoon pogonae (4/18, 22.22%), and Cryptosporidium sp. (1/30, 3.33%). The positivity for protozoa was significantly higher in juveniles compared to adults. Moreover, the frequency of clinical signs was significantly higher in the positive animals. The results obtained here emphasize the importance of regular veterinary examinations of captive P. vitticeps, aimed at the diagnosis, treatment, and control of endoparasites. This study is one of the largest surveys on microsporidia infections in living bearded dragons, suggesting that E. pogonae may be widespread in this lizard. [ABSTRACT FROM AUTHOR]

Protein quality control pathways play important roles in resistance against pathogen infection. For example, the conserved transcription factor SKN-1/NRF up-regulates proteostasis capacity after blockade of the proteasome and also promotes resistance against bacterial infection in the nematode Caenorhabditis elegans. SKN-1/NRF has 3 isoforms, and the SKN-1A/NRF1 isoform, in particular, regulates proteasomal gene expression upon proteasome dysfunction as part of a conserved bounce-back response. We report here that, in contrast to the previously reported role of SKN-1 in promoting resistance against bacterial infection, loss-of-function mutants in skn-1a and its activating enzymes ddi-1 and png-1 show constitutive expression of immune response programs against natural eukaryotic pathogens of C. elegans. These programs are the oomycete recognition response (ORR), which promotes resistance against oomycetes that infect through the epidermis, and the intracellular pathogen response (IPR), which promotes resistance against intestine-infecting microsporidia. Consequently, skn-1a mutants show increased resistance to both oomycete and microsporidia infections. We also report that almost all ORR/IPR genes induced in common between these programs are regulated by the proteasome and interestingly, specific ORR/IPR genes can be induced in distinct tissues depending on the exact trigger. Furthermore, we show that increasing proteasome function significantly reduces oomycete-mediated induction of multiple ORR markers. Altogether, our findings demonstrate that proteasome regulation keeps innate immune responses in check in a tissue-specific manner against natural eukaryotic pathogens of the C. elegans epidermis and intestine. Protein quality control pathways play important roles in resistance against pathogen infection. This study shows that proteasome regulation keeps innate immune responses in check in a tissue-specific manner, against natural eukaryotic pathogens of the C. elegans epidermis and intestine. [ABSTRACT FROM AUTHOR]

(1) Background: Microsporidiosis (nosemosis) is an intestinal disorder of adult honey bees caused by the microsporidian pathogens Vairimorpha apis and Vairimorpha ceranae. In Canada, fumagillin is an approved antibiotic used to treat this disease. However, the recommended dosage is based on efficacy studies for V. apis, the native pathogen in European honey bees. Since the detection of V. ceranae in Apis mellifera, V. ceranae became more prevalent in managed European honey bees and seems to have replaced V. apis due to yet unknown reasons. (2) Methods: This colony study investigated the efficacy of fumagillin administered in the fall to colonies infected with both V. apis and V. ceranae and its effects on the Vairimorpha species' prevalence overwinter. Spore loads in control and fumagillin-treated colonies were analysed by microscopy; Vairimorpha species prevalence was determined molecularly and infection and treatment effects on colony productivity were assessed. (3) Results: Fall fumagillin treatment was associated with a temporary reduction in spore load, but there was no difference in spore loads between treated and control colonies the following spring. Interestingly, fumagillin-treated colonies had a significantly greater prevalence of V. ceranae relative to V. apis the following spring, suggesting fumagillin is less effective in controlling V. ceranae. [ABSTRACT FROM AUTHOR]

Simple Summary: The Vairimorpha ceranae prevalence and the temperature were surveyed and recorded at the National Chung Hsing University apiary for one year. A positive correlation was found between V. ceranae and temperature, suggesting that the prevalence or activity of V. ceranae increases as temperatures rise. Furthermore, the anti-microsporidia activity of the lactic acid bacteria Leuconostoc mesenteroides TBE-8 against V. ceranae was assessed for the management of microsporidiosis. Microsporidiosis, which is caused by the pathogen Vairimorpha ceranae, is a prevalent disease in the honey bee (Apis mellifera) and might lead to significant adult honey bee mortality. In this study, we conducted an annual survey of the mature spore load of V. ceranae in the guts of nurse bees and forager bees in the apiary of National Chung Hsing University (NCHU) in Taiwan. The results indicated that, on average, honey bees hosted approximately 2.13 × 106 mature spore counts (MSCs)/bee in their guts throughout the entire year. The highest number of MSCs was 6.28 × 106 MSCs/bee, which occurred in April 2020, and the lowest number of MSCs was 5.08 × 105 MSCs/bee, which occurred in November 2020. Furthermore, the guts of forager bees had significantly higher (>58%) MSCs than those of nurse bees. To evaluate the potential of the probiotic to treat microsporidiosis, the lactic acid bacterium Leuconostoc mesenteroides TBE-8 was applied to honey bee colonies. A significant reduction (>53%) in MSCs following probiotic treatment was observed, indicating the potential of probiotic treatment for managing microsporidiosis. This research provided information on V. ceranae MSCs in the honey bee gut at NCHU in Taiwan and the MSCs' correlation with the annual season. Furthermore, a potential probiotic treatment for microsporidiosis was assessed for future management. [ABSTRACT FROM AUTHOR]

Porcelain disease, caused by the parasite Astathelohania contejeani, is a fatal disease for freshwater crayfish. Previously published data provide information on the occurrence of A. contejeani in various crayfish species. However, its prevalence in host populations remains largely undetermined. This issue is relevant to the narrow-clawed crayfish Pontastacus leptodactylus, a species of high commercial value in Eastern Europe. A single published report addressed a potential microsporidia infection in P. leptodactylus, but without specific data on geographical location and prevalence. We present the first detailed information on the prevalence of A. contejeani in the P. leptodactylus population from Lake Losvido, including an assessment of the infection rate through both visual and molecular assessment. Astathelohania contejeani was observed in 1.56% of 128 crayfish examined visually. Of the 37 asymptomatic crayfish samples analyzed, 29.7% were found to be carriers. This finding suggests that parasitism of A. contejeani occurs frequently in the P. leptodactylus population in Lake Losvido prior to the manifestation of observable disease symptoms. [ABSTRACT FROM AUTHOR]

During host cell invasion, microsporidian spores translocate their entire cytoplasmic content through a thin, hollow superstructure known as the polar tube. To achieve this, the polar tube transitions from a compact spring-like state inside the environmental spore to a long needle-like tube capable of long-range sporoplasm delivery. The unique mechanical properties of the building blocks of the polar tube allow for an explosive transition from compact to extended state and support the rapid cargo translocation process. The molecular and structural factors enabling this ultrafast process and the structural changes during cargo delivery are unknown. Here, we employ light microscopy and in situ cryo-electron tomography to visualize multiple ultrastructural states of the Vairimorpha necatrix polar tube, allowing us to evaluate the kinetics of its germination and characterize the underlying morphological transitions. We describe a cargo-filled state with a unique ordered arrangement of microsporidian ribosomes, which cluster along the thin tube wall, and an empty post-translocation state with a reduced diameter but a thicker wall. Together with a proteomic analysis of endogenously affinity-purified polar tubes, our work provides comprehensive data on the infection apparatus of microsporidia and uncovers new aspects of ribosome regulation and transport. Microsporidia utilize the polar tube to transport their infectious sporoplasm from the spore to the host for infection. These authors characterize the ultrastructure and composition of the polar tube to identify different states of the tube wall, visualize transported cargo, and discover uniquely arranged ribosomes along the thin tube wall providing new insights into ribosome regulation and transport. [ABSTRACT FROM AUTHOR]

Microsporidia are fungi obligate intracellular pathogens, which infect most animals and cause microsporidiosis. Despite the serious threat that microsporidia pose to humans and agricultural animals, few drugs are available for the treatment and control of microsporidia. To identify novel inhibitors, we took advantage of the model organism Caenorhabditis elegans infected with its natural microsporidian Nematocida parisii. We used this system to screen the Pandemic Response Box, a collection of 400 diverse compounds with known antimicrobial activity. After testing these compounds in a 96-well format at high (100 μM) and low (40 μM) concentrations, we identified four inhibitors that restored the ability of C. elegans to produce progeny in the presence of N. parisii. All four compounds reduced the pathogen load of both N. parisii and Pancytospora epiphaga, a C. elegans-infecting microsporidia related to human-infecting species. One of these compounds, a known inhibitor of a viral protease, MMV1006203, inhibited invasion and prevented the firing of spores. A bis-indole derivative, MMV1593539, decreased spore viability. An albendazole analog, MMV1782387, inhibited proliferation of N. parisii. We tested albendazole as well as 5 other analogs and observed that MMV1782387 was amongst the strongest inhibitors of N. parisii and displayed the least host toxicity. Our study further demonstrates the effectiveness of the C. elegans-N. parisii system for discovering microsporidia inhibitors and the compounds we identified provide potential scaffolds for anti-microsporidia drug development. Author summary: Microsporidia are a large group of parasites which infect both humans and agriculturally important animals such as honey bees and shrimp. These parasites pose a threat to health and food security and there are few therapeutic options available to treat them. To identify compounds that could inhibit microsporidia, we took advantage of the roundworm model organism Caenorhabditis elegans, and the microsporidia Nematocida parisii which naturally infects this worm. We then used this system to screen the Pandemic Response Box, which is a collection of compounds with activity towards pathogens. We identified four compounds which could inhibit microsporidia, and found that these compounds worked by either inactivating microsporidia spores, preventing microsporidia germination, or hindering the growth of the parasite. Together this work provides compounds that could be used to study microsporidia infection and that could be a starting point for the development of drugs against these parasites. [ABSTRACT FROM AUTHOR]

Microsporidia are obligate intracellular parasites that are known to infect most types of animals. Many species of microsporidia can infect multiple related hosts, but it is not known if microsporidia express different genes depending upon which host species is infected or if the host response to infection is specific to each microsporidia species. To address these questions, we took advantage of two species of Nematocida microsporidia, N. parisii and N. ausubeli, that infect two species of Caenorhabditis nematodes, C. elegans and C. briggsae. We performed RNA-seq at several time points for each host infected with either microsporidia species. We observed that Nematocida transcription was largely independent of its host. We also observed that the host transcriptional response was similar when infected with either microsporidia species. Finally, we analyzed if the host response to microsporidia infection was conserved across host species. We observed that although many of the genes upregulated in response to infection are not direct orthologs, the same expanded gene families are upregulated in both Caenorhabditis hosts. Together our results describe the transcriptional interactions of Nematocida infection in Caenorhabditis hosts and demonstrate that these responses are evolutionarily conserved.

Intestinal microsporidiosis is most often caused by Enterocytozoon bieneusi, and to a lesser extent by species of the genus Encephalitozoon. Until now, Encephalitozoon hellem was not clearly known to induce disease restricted to the intestine, or rarely in HIV subjects or in tropical countries. We report here 11 cases of delineated intestinal microsporidioses due to E. hellem diagnosed in France in non-HIV patients. Briefly, all patients were immunocompromised. They all suffered from diarrhoea, associated in nearly 50% of cases with weight loss. Concerning treatment, 5/11 patients had a discontinuation or a decrease of their immunosuppressive therapy, and 4/11 received albendazole. All patients recovered. Five different genotypes were identified based on the rRNA ITS sequence.

Background: The demonstration that the recently discovered Anopheles symbiont Microsporidia MB blocks malaria transmission in Anophelesarabiensis and undergoes vertical and horizontal transmission suggests that it is a promising candidate for the development of a symbiont-based malaria transmission-blocking strategy. The infection prevalence and characteristics of Microsporidia MB in Anopheles gambiae sensu stricto (s.s.), another primary vector species of malaria in Kenya, were investigated. Methods: Field-collected females were confirmed to be Microsporidia MB-positive after oviposition. Egg counts of Microsporidia MB-infected and non-infected individuals were used to infer the effects of Microsporidia MB on fecundity. The time to pupation, adult sex ratio and survival were used to determine if Microsporidia MB infection has similar characteristics in the host mosquitoes An. gambiae and An. arabiensis. The intensity of Microsporidia MB infection in tissues of the midgut and gonads, and in carcasses, was determined by quantitative polymerase chain reaction. To investigate horizontal transmission, virgin males and females that were either Microsporidia MB-infected or non-infected were placed in standard cages for 48 h and allowed to mate; transmission was confirmed by quantitative polymerase chain reaction targeting Microsporidia MB genes. Results: Microsporidia MB was found to naturally occur at a low prevalence in An. gambiae s.s. collected in western Kenya. Microsporidia MB shortened the development time from larva to pupa, but other fitness parameters such as fecundity, sex ratio, and adult survival did not differ between Microsporidia MB-infected and non-infected hosts. Microsporidia MB intensities were high in the male gonadal tissues. Transmission experiments indicated that Microsporidia MB undergoes both maternal and horizontal transmission in An. gambiae s.s. Conclusions: The findings that Microsporidia MB naturally infects, undergoes maternal and horizontal transmission, and is avirulent in An. gambiae s.s. indicate that many of the characteristics of its infection in An. arabiensis hold true for the former. The results of the present study indicate that Microsporidia MB could be developed as a tool for the transmission-blocking of malaria across different Anopheles species. [ABSTRACT FROM AUTHOR]

Microsporidia are a class of obligate intracellular parasitic unicellular eukaryotes that infect a variety of hosts, even including humans. Although different species of microsporidia differ in host range and specificity, they all share a similar infection organelle, the polar tube, which is also defined as the polar filament in mature spores. In response to the appropriate environmental stimulation, the spore germinates with the polar filament everted, forming a hollow polar tube, and then the infectious cargo is transported into host cells via the polar tube. Hence, the polar tube plays a key role in microsporidian infection. Here, we review the origin, structure, composition, function, and application of the microsporidian polar tube, focusing on the origin of the polar filament, the structural differences between the polar filament and polar tube, and the characteristics of polar tube proteins. Comparing the three-dimensional structure of PTP6 homologous proteins provides new insight for the screening of additional novel polar tube proteins with low sequence similarity in microsporidia. In addition, the interaction of the polar tube with the spore wall and the host are summarized to better understand the infection mechanism of microsporidia. Due to the specificity of polar tube proteins, they are also used as the target in the diagnosis and prevention of microsporidiosis. With the present findings, we propose a future study on the polar tube of microsporidia. [ABSTRACT FROM AUTHOR]

Background: The interaction networks between coding and non-coding RNAs (ncRNAs) including long non-coding RNA (lncRNA), covalently closed circular RNA (circRNA) and miRNA are significant to elucidate molecular processes of biological activities and interactions between host and pathogen. Congenital infection caused by vertical transmission of microsporidia N. bombycis can result in severe economic losses in the silkworm-feeding industry. However, little is known about ncRNAs that take place in the microsporidia congenital infection. Here we conducted whole-transcriptome RNA-Seq analyses to identify ncRNAs and regulatory networks for both N. bombycis and host including silkworm embryos and larvae during the microsporidia congenital infection. Results: A total of 4,171 mRNAs, 403 lncRNA, 62 circRNAs, and 284 miRNAs encoded by N. bombycis were identified, among which some differentially expressed genes formed cross-talk and are involved in N. bombycis proliferation and infection. For instance, a lncRNA/circRNA competing endogenous RNA (ceRNA) network including 18 lncRNAs, one circRNA, and 20 miRNAs was constructed to describe 14 key parasites genes regulation, such as polar tube protein 3 (PTP3), ricin-B-lectin, spore wall protein 4 (SWP4), and heat shock protein 90 (HSP90). Regarding host silkworm upon N. bombycis congenital infection, a total of 14,889 mRNAs, 3,038 lncRNAs, 19,039 circRNAs, and 3,413 miRNAs were predicted based on silkworm genome with many differentially expressed coding and non-coding genes during distinct developmental stages. Different species of RNAs form interacting network to modulate silkworm biological processes, such as growth, metamorphosis and immune responses. Furthermore, a lncRNA/circRNA ceRNA network consisting of 140 lncRNAs, five circRNA, and seven miRNAs are constructed hypothetically to describe eight key host genes regulation, such as Toll-6, Serpin-6, inducible nitric oxide synthase (iNOS) and Caspase-8. Notably, cross-species analyses indicate that parasite and host miRNAs play a vital role in pathogen-host interaction in the microsporidia congenital infection. Conclusion: This is the first comprehensive pan-transcriptome study inclusive of both N. bombycis and its host silkworm with a specific focus on the microsporidia congenital infection, and show that ncRNA-mediated regulation plays a vital role in the microsporidia congenital infection, which provides a new insight into understanding the basic biology of microsporidia and pathogen-host interaction. [ABSTRACT FROM AUTHOR]

Microsporidia are a large phylum of intracellular parasites that can infect most types of animals. Species in the Nematocida genus can infect nematodes including Caenorhabditis elegans, which has become an important model to study mechanisms of microsporidia infection. To understand the genomic properties and evolution of nematode-infecting microsporidia, we sequenced the genomes of nine species of microsporidia, including two genera, Enteropsectra and Pancytospora, without any previously sequenced genomes. Core cellular processes, including metabolic pathways, are mostly conserved across genera of nematode-infecting microsporidia. Each species encodes unique proteins belonging to large gene families that are likely used to interact with host cells. Most strikingly, we observed one such family, NemLGF1, is present in both Nematocida and Pancytospora species, but not any other microsporidia. To understand how Nematocida phenotypic traits evolved, we measured the host range, tissue specificity, spore size, and polar tube length of several species in the genus. Our phylogenetic analysis shows that Nematocida is composed of two groups of species with distinct traits and that species with longer polar tubes infect multiple tissues. Together, our work details both genomic and trait evolution between related microsporidia species and provides a useful resource for further understanding microsporidia evolution and infection mechanisms. Author summary: Microsporidia are microbial parasites that can infect many animals. Nematodes have become a useful system to study microsporidia as these animals are commonly infected by microsporidia and these infections can be easily studied in a laboratory environment. To better understand how microsporidia evolve and change their properties as they infect different hosts, we sequenced the genomes of nine microsporidia species which infect nematodes. We found that metabolic pathways were mostly conserved between different clades of these species. Surprisingly, we found a family of proteins predicted to facilitate host interactions that is present in two distinct genera. We also determined the hosts and tissues that these microsporidia can infect as well as morphological properties of the microsporidia. We show that these properties are quite variable in related microsporidia species. Our results allow insight into the evolution of microsporidia and provide a resource for studying microsporidia infections in nematodes. [ABSTRACT FROM AUTHOR]

Water and food sources play a major role in the distribution and transfer of microsporidia infection to animals and humans. So, this systematic review and meta-analysis aimed to assess the status and genetic diversity of microsporidia infection in water, vegetables, fruits, milk, cheese, and meat. The standard protocol of Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines was followed. Scopus, PubMed, Web of Science, and Google Scholar were searched from 1 January 2000 and 1 February 2023. The point estimates and 95% confidence intervals (CIs) were calculated using a random-effects model. Of the 1,308 retrieved studies, 35 articles were included in the final meta-analysis. The pooled prevalence of microsporidia infection in mixed water, mixed fruits, mixed vegetables, and milk was 43.3% (95% CI, 33-54.2%; I2, 94.86%), 35.8% (95% CI, 5.3-84.8%; I2, 0), 12% (95% CI, 4.9-26.6%; I2, 96.43%), and 5.8% (95% CI, 2.7-12%; I2, 83.72%), respectively. Considering the genotypes, microsporidia with genotype D in water sources and genotype CD6 in vegetables/fruits were the highest reported genotypes. Given the relatively high prevalence of microsporidiosis (especially in water sources), designing strategies for control, and prevention of microsporidia infection in these sources should be recommended. [ABSTRACT FROM AUTHOR]

Objectives: We aimed to assess the performance of the Novodiag® Stool Parasites (NSP) assay in the diagnosis of the most common intestinal protozoan and microsporidia infections. Methods: A panel of 167 selected stool samples was retrospectively analysed with the NSP assay and compared to routine microscopy and qPCR methods for the detection of pathogenic protozoa and microsporidia. Results: Whereas specificity was high for all protozoa and microsporidia, NSP sensitivity was strongly dependent on the comparative method used as reference. When compared to microscopic methods, NSP sensitivity was high (96.7 to 100%) for Blastocystis hominis, Entamoeba histolytica and Cyclospora cayetanensis but was lower for Giardia intestinalis (85.2%) and ≤50% for Cystoisospora belli and Dientamoeba fragilis. In comparison to conventional qPCR, the NSP assay demonstrated lower sensitivity characteristics dependent on parasite loads, reaching 60 to 70% for G. intestinalis, D. fragilis, Cryptosporidium spp. and E. histolytica. Sensitivity was 100% for Enterocytozoon bieneusi, but none of the five samples containing Encephalitozoon spp. were detected. Conclusions: The overall performance of the NSP assay in the diagnosis of gastrointestinal protozoa and microsporidia seems to be better than or equivalent to that observed with microscopic methods but inferior to that obtainable with classical targeted qPCR. [ABSTRACT FROM AUTHOR]

Recent declines of wild pollinators and infections in honey, bumble and other bee species have raised concerns about pathogen spillover from managed honey and bumble bees to other pollinators. Parasites of honey and bumble bees include trypanosomatids and microsporidia that often exhibit low host specificity, suggesting potential for spillover to co-occurring bees via shared floral resources. However, experimental tests of trypanosomatid and microsporidial cross-infectivity outside of managed honey and bumble bees are scarce. To characterize potential cross-infectivity of honey and bumble bee-associated parasites, we inoculated three trypanosomatids and one microsporidian into five potential hosts - including four managed species - from the apid, halictid and megachilid bee families. We found evidence of cross-infection by the trypanosomatids Crithidia bombi and C. mellificae, with evidence for replication in 3/5 and 3/4 host species, respectively. These include the first reports of experimental C. bombi infection in Megachile rotundata and Osmia lignaria, and C. mellificae infection in O. lignaria and Halictus ligatus. Although inability to control amounts inoculated in O. lignaria and H. ligatus hindered estimates of parasite replication, our findings suggest a broad host range in these trypanosomatids, and underscore the need to quantify disease-mediated threats of managed social bees to sympatric pollinators.

Microsporidiosis, which is caused by the pathogen Vairimorpha ceranae, is a prevalent disease in the honey bee (Apis mellifera) and might lead to significant adult honey bee mortality. In this study, we conducted an annual survey of the mature spore load of V. ceranae in the guts of nurse bees and forager bees in the apiary of National Chung Hsing University (NCHU) in Taiwan. The results indicated that, on average, honey bees hosted approximately 2.13 × 106 mature spore counts (MSCs)/bee in their guts throughout the entire year. The highest number of MSCs was 6.28 × 106 MSCs/bee, which occurred in April 2020, and the lowest number of MSCs was 5.08 × 105 MSCs/bee, which occurred in November 2020. Furthermore, the guts of forager bees had significantly higher (>58%) MSCs than those of nurse bees. To evaluate the potential of the probiotic to treat microsporidiosis, the lactic acid bacterium Leuconostoc mesenteroides TBE-8 was applied to honey bee colonies. A significant reduction (>53%) in MSCs following probiotic treatment was observed, indicating the potential of probiotic treatment for managing microsporidiosis. This research provided information on V. ceranae MSCs in the honey bee gut at NCHU in Taiwan and the MSCs’ correlation with the annual season. Furthermore, a potential probiotic treatment for microsporidiosis was assessed for future management.

Bats may carry various viruses and bacteria which can be harmful to humans, but little is known about their role as a parasitic source with zoonotic potential. The aim of this study was to test wild bats for the presence of selected parasites: Toxoplasma gondii, Neospora caninum and microsporidia Encephalitozoon spp. In total, brain and small intestine tissues of 100 bats (52 Myotis myotis, 43 Nyctalus noctula and 5 Vespertilio murinus) were used for the DNA isolation and PCR detection of the abovementioned agents. Toxoplasma gondii DNA was detected by real-time PCR in 1% of bats (in one male of M. myotis), while all bats were negative for N. caninum DNA. Encephalitozoon spp. DNA was detected by nested PCR in 25% of bats, including three species (twenty-two M. myotis, two N. noctula and one V. murinus). Positive samples were sequenced and showed homology with the genotypes Encephalitozoon cuniculi II and Encephalitozoon hellem 2C. This is the first study on wild vespertilionid bats from Central Europe and worldwide, with a relatively high positivity of Encephalitozoon spp. detected in bats. [ABSTRACT FROM AUTHOR]

Simple Summary: Microsporidia are obligatory intracellular fungi that cause infection in a broad variety of eukaryotes. However, there are few epidemiological studies on microsporidia in red pandas in China. Therefore, we conducted an epidemiological survey of microsporidia in red pandas from six zoos in Sichuan Province, China. In 198 fecal samples, Enterocytozoon bieneusi was detected in 24 samples (12.1%), while Encephalitozoon spp. was detected in 8 fecal samples (4.0%). Further analysis revealed the presence of genotypes of E. bieneusi with zoonotic concerns. Interestingly, we also observed that the infection rate of microsporidia was higher in semi-free red pandas with more frequent contact with other animals or humans. This is the first report on the occurrence and genotypes of E. bieneusi and Encephalitozoon spp. in red pandas in Sichuan Province, China, contributing to our understanding of the worldwide distribution and genetic diversity of this pathogen. Enterocytozoon bieneusi and Encephalitozoon spp. are microsporidian pathogens with zoonotic potential that pose significant public health concerns. To ascertain the occurrence and genotypes of E. bieneusi and Encephalitozoon spp., we used nested PCR to amplify the internal transcribed spacer (ITS) gene and DNA sequencing to analyze 198 fecal samples from red pandas from 6 zoos in China. The total rate of microsporidial infection was 15.7% (31/198), with 12.1% (24/198), 1.0% (2/198), 2.0% (4/198) and 1.0% (2/198) for infection rate of E. bieneusi, Encephalitozoon cuniculi, Encephalitozoon intestinalis and Encephalitozoon hellem, respectively. One red panda was detected positive for a mixed infection (E. bieneusi and E. intestinalis). Red pandas living in semi-free conditions are more likely to be infected with microsporidia (χ2 = 6.212, df = 1, p < 0.05). Three known (SC02, D, and PL2) and one novel (SCR1) genotypes of E. bieneusi were found. Three genotypes of E. bieneusi (SC02, D, SCR1) were grouped into group 1 with public health importance, while genotype PL2 formed a separate clade associated with group 2. These findings suggest that red pandas may serve as a host reservoir for zoonotic microsporidia, potentially allowing transmission from red pandas to humans and other animals. [ABSTRACT FROM AUTHOR]

Simple Summary: It is well known that mammals can harbor various pathogens that can affect humans (known as zoonotic pathogens) including viruses, bacteria, fungi and parasites. Microsporidia are a group of pathogens related to fungi and parasites of several animals that can cause diarrhea or systemic infection in humans. Due to the limited knowledge about microsporidia infection in hedgehogs worldwide, this study aimed to analyze the presence and identity of microsporidia in a group of North African hedgehogs from the Canary Islands (Spain). Enterocytozoon bieneusi and Encephalitozoon cuniculi, two zoonotic species of microsporidia, were identified. These results suggest that microsporidia species with zoonotic risk circulate in the archipelago. Microsporidia are unicellular eukaryotic obligate intracellular parasites with a wide range of hosts reported worldwide; however, little is known about the epidemiological data on microsporidia infection in animals from the Canary Islands. Since data on microsporidia infection in hedgehog species are scarce, the aim of this study was to analyze the presence and identity of microsporidia in a group of North African hedgehogs (Atelerix algirus) using microscopic and molecular methods. From December 2020 to September 2021, a total of 36 fecal samples were collected from naturally deceased hedgehogs from Tenerife and Gran Canaria. All samples showed spore-compatible structures (100%; 36/36) under microscopic analysis, of which 61.1% (22/36) were amplified via the nested-polymerase chain reaction (PCR) targeting the partial sequence of the 16S rRNA gene, the internal transcribed spacer (ITS) region, and the partial sequence of the 5.8S rRNA gene. After Sanger sequencing and ITS analysis, Enterocytozoon bieneusi was detected in 47.2% (17/36) of the samples, identifying two novel genotypes (AAE1 and AAE2), followed by the detection of an undetermined species in 8.3% (3/36) and Encephalitozoon cuniculi genotype I in 5.6% (2/36) of the samples. This study constitutes the first report of microsporidia species in Atelerix algirus worldwide, highlighting the high prevalence of zoonotic species. [ABSTRACT FROM AUTHOR]

Introduction: Nosema is a diverse genus of unicellular microsporidian parasites of insects and other arthropods. Nosema muscidifuracis infects parasitoid wasp species of Muscidifurax zaraptor and M. raptor (Hymenoptera: Pteromalidae), causing ~50% reduction in longevity and ~90% reduction in fecundity. Methods and Results: Here, we report the first assembly of the N. muscidifuracis genome (14,397,169bp in 28 contigs) of high continuity (contig N50 544.3 Kb) and completeness (BUSCO score 97.0%). A total of 2,782 protein-coding genes were annotated, with 66.2% of the genes having two copies and 24.0% of genes having three copies. These duplicated genes are highly similar, with a sequence identity of 99.3%. The complex pattern suggests extensive gene duplications and rearrangements across the genome. We annotated 57 rDNA loci, which are highly GC-rich (37%) in a GC-poor genome (25% genome average). Nosema-specific qPCR primer sets were designed based on 18S rDNA annotation as a diagnostic tool to determine its titer in host samples. We discovered high Nosema titers in Nosemacured M. raptor and M. zaraptor using heat treatment in 2017 and 2019, suggesting that the remedy did not completely eliminate the Nosema infection. Cytogenetic analyses revealed heavy infections of N. muscidifuracis within the ovaries of M. raptor and M. zaraptor, consistent with the titer determined by qPCR and suggesting a heritable component of infection and per ovum vertical transmission. Discussion: The parasitoids-Nosema system is laboratory tractable and, therefore, can serve as a model to inform future genome manipulations of Nosema-host system for investigations of Nosemosis. [ABSTRACT FROM AUTHOR]

Featured Application: Microsporidiosis leads to the formation of dead intestine cell spots, which can cause interruption of the host's intestinal continuity and lead to intestinal leaking. Background: Microsporidia from the Nosema (Vairimorpha) genus are pathogenic fungi that complete their life cycle in the honeybee intestine. Therefore, the aim of this study was to determine the impact of the course of infection on the viability of honeybee intestine cells. Methods and Results: Intestines isolated from healthy and N. ceranae-infected honeybees were stained using two dyes, SYTO 9 and propidium iodide, and analyzed under an Axiovert 200M fluorescence microscope immediately after the isolation of the intestines. The ImageJ program was used for the quantitative analysis of the cell structure parameters. Our study demonstrated for the first time that healthy bees have a higher number of live cells in their intestines than infected bees, and that the intestines of N. ceranae-infected honeybees contain dead cells concentrated in spots. The results obtained for these two cases differed significantly, and were confirmed by statistical tests. Conclusions: The intestines of infected honeybees contain dead cells concentrated in red/dead spots, which can lead to necrotic changes, the interruption of the host's intestinal continuity, intestinal leaking and the increased mortality of the host. [ABSTRACT FROM AUTHOR]

Simple Summary: Microsporidia are intracellular parasites that have detrimental effects on their various invertebrate and vertebrate hosts. The microsporidia pathogenic to honeybees and bumblebees belong to the genus Vairimorpha (previously Nosema). Among these, V. bombi is the most widespread species worldwide and it infects a variety of bumblebee species. In this study, we investigated the infection of V. bombi among Japanese native bumblebees and the alien species, Bombus terrestris, established in Japan, using PCR and microscopy. We discovered that V. bombi infected the subgenus Bombus s. str. species/subspecies with a low prevalence in areas both with and without the presence of B. terrestris, suggesting that V. bombi may be native to Japan. In contrast, a newly discovered microsporidian parasite that is phylogenetically distinct from V. bombi was found to infect the subgenus Diversobombus species/subspecies with a high prevalence. Hence, if the new microsporidian species does not negatively affect the host and is vertically transmitted from queen to offspring, it may coexist with its host. In addition, this may suggest that V. bombi and the new microsporidium have different life history strategies. Microsporidia are spore-forming intracellular parasites of various invertebrates and vertebrates. Vairimorpha bombi negatively affects the fitness of bumblebees and its prevalence correlates with declining bumblebee populations. The invasive alien species Bombus terrestris colonized Japan and possibly introduced new parasites. To assess the infection prevalence of V. bombi in Japanese bumblebees and B. terrestris, we investigated V. bombi infections using PCR and microscopy. The prevalence of sporulating V. bombi infections in three Bombus s. str. species/subspecies was low, whereas that of non/low-sporulating Vairimorpha sp. infections in three Diversobombus species/subspecies was high. Invasive B. terrestris showed low prevalence of non/low-sporulating V. bombi infections and shared the same V. bombi haplotype with B. hypocrita found in Hokkaido, where B. terrestris is present, and in Honshu, where B. terrestris is absent. Although V. bombi may have been introduced with B. terrestris colonies imported from Europe, it seems to be originally distributed in Japan. Furthermore, a new Vairimorpha sp. was found in Japanese bumblebee species. V. bombi and Vairimorpha sp. showed different organ and host specificities in bumblebees. There are no reports on the specific effects of other Vairimorpha spp. on bumblebees; further studies are needed to clarify the individual characteristics of Vairimorpha spp. [ABSTRACT FROM AUTHOR]

We report a case of Anncaliia algerae microsporidia infection in an immunosuppressed kidney transplant recipient in China. Light microscopy and transmission electron microscopy initially failed to identify A. algerae, which eventually was detected by metagenomic next-generation sequencing. Our case highlights the supporting role of metagenomic sequencing in early identification of uncommon pathogens.

Animals are under constant selective pressure from a myriad of diverse pathogens. Microsporidia are ubiquitous animal parasites, but the influence they exert on shaping animal genomes is mostly unknown. Using multiplexed competition assays, we measured the impact of four different species of microsporidia on 22 wild isolates of Caenorhabditis elegans. This resulted in the identification and confirmation of 13 strains with significantly altered population fitness profiles under infection conditions. One of these identified strains, JU1400, is sensitive to an epidermal-infecting species by lacking tolerance to infection. JU1400 is also resistant to an intestinal-infecting species and can specifically recognize and destroy this pathogen. Genetic mapping of JU1400 demonstrates that these two opposing phenotypes are caused by separate loci. Transcriptional analysis reveals the JU1400 sensitivity to epidermal microsporidia infection results in a response pattern that shares similarity to toxin-induced responses. In contrast, we do not observe JU1400 intestinal resistance being regulated at the transcriptional level. The transcriptional response to these four microsporidia species is conserved, with C. elegans strain-specific differences in potential immune genes. Together, our results show that phenotypic differences to microsporidia infection amongst C. elegans are common and that animals can evolve species-specific genetic interactions. Author summary: Animals display a large diversity of different characteristics within a population. One characteristic that varies within populations is susceptibility to infectious disease. Microsporidia are ubiquitous animal parasites that infect most types of animals, but the influence that these parasites have had on animal evolution and diversity is not well understood. Using wild isolates of the model organism Caenorhabditis elegans, we tested the diversity of responses to infection by four naturally occurring microsporidia species. These experiments were done with a pooled assay that allowed for the determination of the relative abundance of strains within a population under different infection conditions. These experiments identified a C. elegans strain that is sensitive to infection by one species of microsporidia but resistant to another. Genetic mapping experiments show that different genetic regions are responsible for these different infection responses. Together our study demonstrates that there is variability in the susceptibility of C. elegans to microsporidia infection and suggests that these parasites have likely had a large influence on animal evolution. [ABSTRACT FROM AUTHOR]

Aquaculture combines techniques for breeding and harvesting aquatic organisms used in shrimp production. It is important as a source of income and for generating foreign exchange in the regions where it is practiced. However, the timely detection of diseases continues to be a great challenge for aquaculture and fisheries production. In recent years, Enterocytozoon hepatopenaei (EHP) has emerged as a major pathogen of the Pacific white shrimp Penaeus vannamei in many Asian countries (Vietnam, China, Indonesia, Malaysia, Thailand, India, and Korea). In Latin America, only in Venezuela, and to date, there is no report of its presence in Mexico. It is transmitted directly from shrimp to shrimp by oral or fecal means, cannibalism, or exposure to contaminated water. Hepatopancreatic microsporidiosis (HPM) is mainly associated with stunted growth and severe infections that can cause a poor production cycle, mortality, and problems in larva-producing laboratories. This review aims to overview the main microsporidian parasites and diseases found in white shrimp, including the clinical signs, control and prevention measures for EHP infection, and the detection of HPM using different techniques. In order to offer timely detection tools, different techniques are available for the detection and study of microsporidia. Such as optical microscopy, transmission electron microscopy, and histology; however, for diagnostic purposes, molecular methods are preferred due to their sensitivity, specificity, and short-time analysis. Our review suggests that constant monitoring in shrimp hatcheries and farms is essential to avoid the entry or transference of infected organisms, affecting shrimp production and the ideal development of healthy shrimp. [ABSTRACT FROM AUTHOR]

Introduction: Enterocytozoon bieneusi (E. bieneusi) Microsporidia can cause opportunistic infections in immunocompromised patients and is also an emerging disease in these individuals. Its clinical manifestations are chronic diarrhea and severe wasting syndrome, these can be extremely debilitating and carry a significant risk of death for immunocompromised patients. Often, microsporidia cannot be confirmed immediately by routine examination and culture. Effective and available treatment options are limited for infections caused by E. bieneusi in humans. Such cases are very rare in Chinese Mainland. Case presentation: A 47-year-old male had recurrent, profuse watery diarrhea and abdominal discomfort for more than 7 months, with a fever for 5 days. Two years earlier, he received treatment with amodified BFM-90 protocol for acute B cell lymphoblastic leukemia and is currently in the final stages of maintenance therapy with oralmethotrexate and mercaptopurine. The leukemia was assessed as still in remission two months ago. PET/CT showed massive peritoneal fluid accumulation and a high uptake area in the diffused peritoneum (SUVmax 12.57), suggesting tumor invasion or microbial infections. However, broad-spectrum antibacterial therapies were ineffective. Metagenomic sequencing of plasma and peritoneal fluid showed no suggestion of the existence of a tumor but instead showed a high sequence number of DNA and RNA of the Microsporidia. His albendazole treatment failed and subsequent treatment with nitazoxanide successfully resolved the infection. Conclusion: This case shows that we should consider the possibility of atypical pathogen infection in patients with hematologic malignancy who repeatedly develop unexplained diarrhea with wasting. mNGS can help rule out malignant neoplasms and diagnose infections. Our results suggest that nitazoxanide effectively treats E. bieneusi microsporidia infections. [ABSTRACT FROM AUTHOR]

Microsporidiosis is an emerging opportunistic human infection, capable of producing from a localized infection to a systemic infection. The objective was to detect the presence of Microsporidia in samples from HIV positive patients with diarrheal syndrome at the "Dr. José Daniel Rodríguez" from Guayaquil, 2021-2022. 87 patients were studied using the optical microscopy method, with fast-hot Gram Chromotrope staining, the analysis of results was carried out through descriptive statistics and Excel and SPSS software were used. About the total number of individuals studied, 13 (14.9%) patients were found positive for Microsporidia and 74 (85.1%) did not present spores. Regarding the level of education, 8 (61.5%) have primary school and 5 have secondary school (38.5%), marital status 10 (76.9%) are single and 3 (23.1%) are married, the highest concentration is in Guayaquil 8 (6.5%), Durán 1 (7.7%), Esmeralda 1 (7.7%) and Riobamba 3 (23.1%). The levels of LTCD4+, 7 (53.85%) patients had below 500 xml3 cells and 6 (46.15%) had more than 500 To the type of diarrhea, 13 (100%) presented chronic diarrhea. In conclusion, the optical microscopy method using fast-hot Gram Chromotrope staining allowed the detection of microsporidia spores, demonstrating high sensitivity and specificity. [ABSTRACT FROM AUTHOR]

Obligately intracellular microsporidia regulate their host cell life cycles, including apoptosis, but this has not been evaluated in phagocytic host cells such as macrophages that can facilitate infection but also can be activated to kill microsporidia. We examined two biologically dissimilar human-infecting microsporidia species, Encephalitozoon cuniculi and Vittaforma corneae, for their effects on staurosporine-induced apoptosis in the human macrophage-differentiated cell line, THP1. Apoptosis was measured after exposure of THP-1 cells to live and dead mature organisms via direct fluorometric measurement of Caspase 3, colorimetric and fluorometric TUNEL assays, and mRNA gene expression profiles using Apoptosis RT2 Profiler PCR Array. Both species of microsporidia modulated the intrinsic apoptosis pathway. In particular, live E. cuniculi spores inhibited staurosporine-induced apoptosis as well as suppressed pro-apoptosis genes and upregulated anti-apoptosis genes more broadly than V. corneae. Exposure to dead spores induced an opposite effect. Vittaforma corneae, however, also induced inflammasome activation via Caspases 1 and 4. Of the 84 apoptosis-related genes assayed, 42 (i.e. 23 pro-apoptosis, nine anti-apoptosis, and 10 regulatory) genes were more affected including those encoding members of the Bcl2 family, caspases and their regulators, and members of the tumour necrosis factor (TNF)/TNF receptor R superfamily.

Natural genetic variation can determine the outcome of an infection, and often reflects the co-evolutionary battle between hosts and pathogens. We previously found that a natural variant of the nematode Caenorhabditis elegans from Hawaii (HW) has increased resistance against natural microsporidian pathogens in the Nematocida genus, when compared to the standard laboratory strain of N2. In particular, HW animals can clear infection, while N2 animals cannot. In addition, HW animals have lower levels of initial colonization of Nematocida inside intestinal cells, compared to N2. Here we investigate how this natural variation in resistance relates to autophagy. We found that there is much better targeting of autophagy-related machinery to parasites under conditions where they are cleared. In particular, ubiquitin targeting to Nematocida cells correlates very well with their subsequent clearance in terms of timing, host strain and age, as well as species of Nematocida. Furthermore, clearance correlates with targeting of the LGG-2/LC3 autophagy protein to parasite cells, with HW animals having much more efficient targeting of LGG-2 to parasite cells than N2 animals. Surprisingly, however, we found that LGG-2 is not required to clear infection. Instead, we found that LGG-2/LC3 regulates Nematocida colonization inside intestinal cells. Interestingly, LGG-2/LC3 regulates intracellular colonization only in the HW strain, and not in N2. Altogether these results demonstrate that there is natural genetic variation in an LGG-2-dependent process that regulates microsporidia colonization inside intestinal cells, although not microsporidia clearance.

Simple Summary: Microsporidia are a group of fungal-related pathogens widely distributed in the environment, with some species having a negative impact on animals and public health. The European rabbit (Oryctolagus cuniculus) is considered the natural host of Encephalitozoon cuniculi, a microsporidian pathogen of mammals, including humans. The infection caused by E. cuniculi, encephalitozoonosis, ranges from asymptomatic to severe lesions in rabbits, with clinical signs involving the central nervous system, kidney, and eye being the most common. The majority of reported cases have been in domestic rabbits, while cases in wild rabbits are uncommon. Due to the lack of data on microsporidia in the Canary Islands, the aim of this work was to analyze the prevalence and identity of microsporidia in fecal samples from rabbits in Tenerife. Enterocytozoon bieneusi and Encephalitozoon spp. are microsporidia with zoonotic potential that have been identified in humans, as well as in a large group of wild and domestic animals. Several wildlife species have been studied as reservoirs of zoonotic microsporidia in mainland Spain, including the European rabbit (Oryctolagus cuniculus). Due to a lack of data on microsporidia infection in wildlife on the Canary Islands, the aim of this work was to analyze the prevalence and identify the species of microsporidia in rabbits in Tenerife. Between 2015 and 2017, a total of 50 fecal samples were collected from rabbits in eight municipalities of Tenerife, Canary Islands, Spain. Seven of the fifty samples (14%) were amplified using nested polymerase chain reaction (PCR) targeting the partial sequence of the 16S rRNA gene, the internal transcribed spacer (ITS) region, and the partial sequence of the 5.8S rRNA gene. Sanger sequencing reveals the presence of Encephalitozoon cuniculi genotype I in two samples (4%), and undescribed microsporidia species in five samples (10%). This study constitutes the first molecular detection and genotyping of E. cuniculi in rabbits in Spain. [ABSTRACT FROM AUTHOR]

Nosemosis is the most common disease in honey bee Apis mellifera L., and is a major issue related to bee health worldwide. Therefore, the purpose of this research study was to determine prevalence of microsporidia parasitic infection of the genus Nosema spp. in East Kazakhstan Region (EKR). In the years of 2018 -2021, 394 honey bee samples were collected at 30 apiaries located in four districts of East Kazakhstan Region (Katon-Karagay, Urzhar, Borodulikhinsky, and Shemonaikhinsky). In order to determine the level of infestation, firstly, the presence of Nosema spp. spores was detected using optical microscopy, and then the average amount of spores per bee was counted using a hemocytometer. The degree of nosemosis prevalence was determined in points by means of a semi-quantitative method, and as a percentage from the total of samples and of the amount of positive tests. At the outcome of the study, microsporidia of the genus Nosema spp. were detected in 23.3% of cases (92 samples). Prevalence at its low degree was found in six samples (1.5%), at an average degree in 55 samples (14%), and at a high one in 31 samples (7.9%). This research study proved that microsporidia of the genus Nosema spp. are widely spread at the apiaries of East Kazakhstan Region in different orographic and climatic conditions. Notwithstanding that it was impossible to statistically determine any significant differences between the dependence of nosemosis prevalence and the apiary location, this indicator is actually higher in the mountainous regions than in the steppe. Concurrently, a close inverse correlation was recognized between the amount of spores in one bee and the level of infestation in bee families from the duration of the vegetation season at the apiary location. This gives grounds to assert that the environmental factors have an impact on formation and development of nosemosis. The results of the research presented in the article indicate the need for further research aimed at increasing the number of studied apiaries, and above all the use of molecular biology methods to distinguish the species that cause nosemosis infection (PCR). [ABSTRACT FROM AUTHOR]

Regulation of immunity throughout an organism is critical for host defense. Previous studies in the nematode Caenorhabditis elegans have described an "ON/OFF" immune switch comprised of the antagonistic paralogs PALS-25 and PALS-22, which regulate resistance against intestinal and epidermal pathogens. Here, we identify and characterize a PALS-25 gain-of-function mutant protein with a premature stop (Q293*), which we find is freed from physical repression by its negative regulator, the PALS-22 protein. PALS-25(Q293*) activates two related gene expression programs, the Oomycete Recognition Response (ORR) against natural pathogens of the epidermis, and the Intracellular Pathogen Response (IPR) against natural intracellular pathogens of the intestine. A subset of ORR/IPR genes is upregulated in pals-25(Q293*) mutants, and they are resistant to oomycete infection in the epidermis, and microsporidia and virus infection in the intestine, but without compromising growth. Surprisingly, we find that activation of PALS-25 seems to primarily stimulate the downstream bZIP transcription factor ZIP-1 in the epidermis, with upregulation of gene expression in both the epidermis and in the intestine. Interestingly, we find that PALS-22/25-regulated epidermal-to-intestinal signaling promotes resistance to the N. parisii intestinal pathogen, demonstrating cross-tissue protective immune induction from one epithelial tissue to another in C. elegans. Author summary: Multicellular organisms need to monitor the health and function of multiple tissues simultaneously to respond appropriately to pathogen infection. Here, we study an ON/OFF switch in the roundworm C. elegans that controls immune responses to diverse natural pathogens of the skin and gut. We show a physical association between the 'ON switch' protein PALS-25 and the 'OFF switch' protein PALS-22, and that this association is disrupted in a mutant, activated form of PALS-25. When either PALS-22 is lost, or PALS-25 is activated, a downstream immune regulator ZIP-1 is activated more prominently in the skin than in the gut. Excitingly, our findings show that skin-specific loss of PALS-22 or skin-specific activation of PALS-25 can induce immune responses in the worm gut. These findings highlight the coordination of immune responses across different tissues that are commonly infected by microbial pathogens. [ABSTRACT FROM AUTHOR]

Simple Summary: The Iberian lynx, which inhabits the Iberian Peninsula, is one of the most endangered felines in the world. Wild Iberian lynx populations have suffered a constant regression over the past century, with a rapid decline of 90% in the last 20 years. Infectious diseases are one of the most critical threats that cause the population decline of these animals, either in the wild or captivity. Different studies have revealed positive seroprevalence against various pathogens, confirming contact and exposure to bacteria, viruses, and parasites. In this sense, searching for pathogens related to the depopulation of the Iberian lynx is vital for conserving and maintaining this threatened species. The present work confirmed the presence of microsporidia, opportunistic intracellular parasites recently related to fungi, in the lynx environment. Also, different species of microsporidia were determined for the first time in the urine, feces, and tissue samples of Lynx pardinus. Further studies are needed to establish the impact of microsporidia infection on the survival of the Iberian lynx. These studies would contribute to the endurance and conservation of this feline by implementing new prevention strategies. Lynx pardinus is one of the world's most endangered felines inhabiting the Iberian Peninsula. The present study was performed to identify the presence of microsporidia due to the mortality increase in lynxes. Samples of urine (n = 124), feces (n = 52), and tissues [spleen (n = 13), brain (n = 9), liver (n = 11), and kidney (n = 10)] from 140 lynxes were studied. The determination of microsporidia was evaluated using Weber's chromotrope stain and Real Time-PCR. Of the lynxes analyzed, stains showed 10.48% and 50% positivity in urine and feces samples, respectively. PCR confirmed that 7.69% and 65.38% belonged to microsporidia species. The imprints of the tissues showed positive results in the spleen (38.46%), brain (22.22%), and liver (27.27%), but negative results in the kidneys. PCR confirmed positive microsporidia results in 61.53%, 55.55%, 45.45%, and 50%, respectively. Seroprevalence against Encephalitozoon cuniculi was also studied in 138 serum samples with a positivity of 55.8%. For the first time, the results presented different species of microsporidia in the urine, feces, and tissue samples of Lynx pardinus. The high titers of anti-E. cuniculi antibodies in lynx sera confirmed the presence of microsporidia in the lynx environment. New studies are needed to establish the impact of microsporidia infection on the survival of the Iberian lynx. [ABSTRACT FROM AUTHOR]

Fruit black spot (FBS), a fungal disease of pecan (Carya illinoinensis (Wangenh) K. Koch) caused by the pathogen Pestalotiopsis microspora, is a serious disease and poses a critical threat to pecan yield and quality. However, the details of pecan responses to FBS infection at the transcriptional level remain to be elucidated. In present study, we used RNA-Seq to analyze differential gene expression in three pecan cultivars with varied resistance to FBS infection: Xinxuan-4 (X4), Mahan (M), and Wichita (W), which were categorized as having low, mild, and high susceptibility to FBS, respectively. Nine RNA-Seq libraries were constructed, comprising a total of 58.56 Gb of high-quality bases, and 2420, 4380, and 8754 differentially expressed genes (DEGs) with |log2Fold change| ≥ 1 and p-value < 0.05 were identified between M vs. X4, W vs. M, and W vs. X4, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathway analyses were performed to further annotate DEGs that were part of specific pathways, which revealed that out of 134 total pathways, MAPK signaling pathway, plant–pathogen interaction, and plant hormone signal transduction were highly enriched. Transcriptomic profiling analysis revealed that 1681 pathogen-related genes (PRGs), including 24 genes encoding WRKY transcription factors, potentially participate in the process of defense against Pestalotiopsis microspora infection in pecan. The correlation of WRKY TFs and PRGs was also performed to reveal the potential interaction networks among disease-resistance/pathogenesis-related genes and WRKY TFs. Expression profiling of nine genes annotated as TIFY, WRKY TF, and disease-resistance protein-related genes was performed using qRT-PCR, and the results were correlated with RNA-Seq data. This study provides valuable information on the molecular basis of pecan–Pestalotiopsis microspora interaction mechanisms and offers a repertoire of candidate genes related to pecan fruit response to FBS infection. [ABSTRACT FROM AUTHOR]

We identified Encephalitozoon cuniculi genotype II parasites as a cause of extraintestinal microsporidiosis in 2 owners of birds also infected with E. cuniculi. Patients experienced long-lasting nonspecific symptoms; the disease course was more progressive in a patient with diabetes. Our findings suggest direct bird-to-human transmission of this pathogen.

Abstract Background Microsporidia is a large group of eukaryotic obligate intracellular spore-forming parasites, of which 17 species can cause microsporidiosis in humans. Most human-infecting microsporidians belong to the genera Enterocytozoon and Encephalitozoon. To date, only five microsporidian species, including Encephalitozoon-like, have been found in hard ticks (Ixodidae) using microscopic methods, but no sequence data are available for them. Furthermore, no widespread screening for microsporidian-infected ticks based on DNA analysis has been carried out to date. Thus, in this study, we applied a recently developed DNA metabarcoding method for efficient microsporidian DNA identification to assess the role of ticks as potential vectors of microsporidian species causing diseases in humans. Methods In total, 1070 (493 juvenile and 577 adult) unfed host-seeking Ixodes ricinus ticks collected at urban parks in the city of Poznan, Poland, and 94 engorged tick females fed on dogs and cats were screened for microsporidian DNA. Microsporidians were detected by PCR amplification and sequencing of the hypervariable V5 region of 18S rRNA gene (18S profiling) using the microsporidian-specific primer set. Tick species were identified morphologically and confirmed by amplification and sequencing of the shortened fragment of cytochrome c oxidase subunit I gene (mini-COI). Results All collected ticks were unambiguously assigned to I. ricinus. Potentially zoonotic Encephalitozoon intestinalis was identified in three fed ticks (3.2%) collected from three different dogs. In eight unfed host-seeking ticks (0.8%), including three males (1.1%), two females (0.7%) and three nymphs (0.7%), the new microsporidian sequence representing a species belonging to the genus Endoreticulatus was identified. Conclusions The lack of zoonotic microsporidians in host-seeking ticks suggests that I. ricinus is not involved in transmission of human-infecting microsporidians. Moreover, a very low occurrence of the other microsporidian species in both fed and host-seeking ticks implies that mechanisms exist to defend ticks against infection with these parasites. Graphical abstract

Microsporidia are a diverse group of fungal-related obligate intracellular parasites that infect most animal phyla. Despite the emerging threat that microsporidia represent to humans and agricultural animals, few reliable treatment options exist. Here, we develop a high-throughput screening method for the identification of chemical inhibitors of microsporidia infection, using liquid cultures of Caenorhabditis elegans infected with the microsporidia species Nematocida parisii. We screen a collection of 2560 FDA-approved compounds and natural products, and identify 11 candidate microsporidia inhibitors. Five compounds prevent microsporidia infection by inhibiting spore firing, whereas one compound, dexrazoxane, slows infection progression. The compounds have in vitro activity against several other microsporidia species, including those known to infect humans. Together, our results highlight the effectiveness of C. elegans as a model host for drug discovery against intracellular pathogens, and provide a scalable high-throughput system for the identification and characterization of microsporidia inhibitors. Microsporidia are fungal-related intracellular parasites that infect animals and humans. Here, Murareanu et al. develop a high-throughput screening method using the nematode C. elegans as a host, and identify several compounds that inhibit microsporidia infection through different mechanisms. [ABSTRACT FROM AUTHOR]

Background: Microsporidia are a group of obligated intracellular fungus pathogens. Monocytes and the derivative macrophages are among the most important players in host immunity. The invasion of microsporidia may significantly affect the monocytes maturation and extravasation processes. Methods: We utilized a previously established microsporidia infection murine model to investigate the influences of microsporidia Encephalitozoon hellem (E. hellem) infection on monocyte maturation, releasing into the circulation and extravasation to the inflammation site. Flow cytometry and qPCR analysis were used to compare the monocytes and derivative macrophages isolated from bone marrow, peripheral blood and tissues of E. hellem-infected and control mice. Results: The results showed that the pro-inflammatory group of CD11b+Ly-6C+ monocytes are promoted in E. hellem-infected mice. Interestingly, the percentage of Ly-6C+ monocytes from E. hellem-infected mice are significantly lower in peripheral blood while significantly higher in the inflamed small intestine, together with up-regulated ratio of F4/80 macrophage in small intestine as well. Conclusions: Our findings demonstrated that E. hellem infection leads to promoted monocytes maturation in bone marrow, up-regulation of extravasation from peripheral blood to inflammation site and maturation into macrophages. Our study is the first systematic analysis of monocytes maturation and trafficking during microsporidia infection, and will provide better understanding of the pathogen–host interactions. [ABSTRACT FROM AUTHOR]

Bats may carry various viruses and bacteria which can be harmful to humans, but little is known about their role as a parasitic source with zoonotic potential. The aim of this study was to test wild bats for the presence of selected parasites: Toxoplasma gondii, Neospora caninum and microsporidia Encephalitozoon spp. In total, brain and small intestine tissues of 100 bats (52 Myotis myotis, 43 Nyctalus noctula and 5 Vespertilio murinus) were used for the DNA isolation and PCR detection of the abovementioned agents. Toxoplasma gondii DNA was detected by real-time PCR in 1% of bats (in one male of M. myotis), while all bats were negative for N. caninum DNA. Encephalitozoon spp. DNA was detected by nested PCR in 25% of bats, including three species (twenty-two M. myotis, two N. noctula and one V. murinus). Positive samples were sequenced and showed homology with the genotypes Encephalitozoon cuniculi II and Encephalitozoon hellem 2C. This is the first study on wild vespertilionid bats from Central Europe and worldwide, with a relatively high positivity of Encephalitozoon spp. detected in bats.

Emerging infectious disease has become the center of attention since the outbreak of COVID-19. For the coronavirus, bats are suspected to be the origin of the pandemic. Consequently, the spotlight has fallen on zoonotic diseases, and the focus now expands to organisms other than viruses. Microsporidia is a single-cell organism that can infect a wide range of hosts such as insects, mammals, and humans. Its pathogenicity differs among species, and host immunological status plays an important role in infectivity and disease severity. Disseminated disease from microsporidiosis can be fatal, especially among patients with a defective immune system. Recently, there were two Trachipleistophora hominis , a microsporidia species which can survive in insects, case reports in Thailand, one patient had disseminated microsporidiosis. This review gathered data of disseminated microsporidiosis and T. hominis infections in humans covering the biological and clinical aspects. There was a total of 22 cases of disseminated microsporidiosis reports worldwide. Ten microsporidia species were identified. Maximum likelihood tree results showed some possible correlations with zoonotic transmissions. For T. hominis, there are currently eight case reports in humans, seven of which had Human Immunodeficiency Virus (HIV) infection. It is observed that risks are higher for the immunocompromised to acquire such infections, however, future studies should look into the entire life cycle, to identify the route of transmission and establish preventive measures, especially among the high-risk groups. [ABSTRACT FROM AUTHOR]

Here, we report, for the first time, a microsporidian infection in laboratory-reared larvae of the damselfly Ischnura elegans. Infected larvae originated from field-collected adult females, which were caught in southern Poland in August 2020 (the second half of the flight season). Higher rearing temperatures and the presence of predator cues from the invasive alien signal crayfish (Pacifastacus leniusculus) increased the number of infected larvae. Infected larvae had distorted wing development, and all individuals died before emergence. Hence, microsporidian infection in I. elegans larvae impacted damselfly morphology and life history. We propose that warming temperature and stress caused by non-consumptive effects triggered by invasive alien predators are possible factors that produce negative fitness consequences following microsporidian infection in a key amphibious ectotherm. [ABSTRACT FROM AUTHOR]

Paromomycin is a naturally occurring aminoglycoside antibiotic that has effects on both prokaryotic and eukaryotic microbes. However, previous reports have indicated that it has little effect on microsporidia, including Vairimorpha (Nosema) ceranae, in cell culture models. V. ceranae is one of a number of microsporidia species that cause disease in honey bees and substantial efforts to find new treatment strategies for bees that are infected with these pathogens are ongoing. When testing compounds for potential activity against V. ceranae in whole organisms, we found that paromomycin reduces the infection intensity of this parasite. Critically, the necessary doses of paromomycin have high activity against the bacteria of the honey bee microbiome and cause evident stress in bees. Microsporidia have been shown to lack an essential binding site on the ribosome that is known to allow for maximal inhibition by paromomycin. Thus, it is possible that paromomycin impacts parasite levels through non-cell autonomous effects on microsporidia infection levels via effects on the microbiome or midgut cellular function. As paromomycin treatment could cause widespread honey bee health issues in agricultural settings, it does not represent an appropriate anti-microsporidia agent for use in the field. [ABSTRACT FROM AUTHOR]

Background: Microsporidia are spore‐forming intracellular pathogens with worldwide prevalence, causing emerging infections in humans and animals. Enterocytozoon bieneusi is a zoonotic species of microsporidia and is responsible for more than 90% of cases of microsporidiosis in humans and animals. Pigs and wild boars are important animal reservoirs of microsporidia. Hence, we aimed to estimate the global prevalence of microsporidia and genetic diversity of E. bieneusi in pigs and wild boars through a set of systematic review and meta‐analysis (PRISMA) guidelines. Methods: Four databases (Web of Science, PubMed, Scopus and Google Scholar) were searched between January 1, 2000 and April 30, 2021. Regarding meta‐analysis, the random‐effect model was employed by forest plot with 95% confidence interval (CI). Results: After exclusion of irrelevant articles and duplication removal, 33 papers, including 34 datasets (30 datasets for domestic pigs and 4 for wild boars) finally meet the inclusion criteria to undergo meta‐analysis. The pooled prevalence rates of microsporidia infection in domestic pigs and wild boars were 37.6% (95% CI: 30.8–44.9%) and 8.1% (95% CI: 2.1–26.8%), respectively. While, the pooled prevalence rates of E. bieneusi were 35% (95% CI: 28.4–42.2%) in domestic pigs and 10.1% (95% CI: 1.7–42.4%) in wild boars. The genotypes EbpA was the most reported genotype in domestic pigs and wild boars. Male animals had higher prevalence rates of microsporidia infection than females (27 vs. 17.4%, OR = 1.91; 95% CI, 0.77–4.71%). Conclusion: This study indicates the important role of domestic pigs and wild boars as animal reservoir hosts of microsporidia. Thereby, strategies for control and prevention of these zoonotic pathogens should be designed in pigs and wild boars. [ABSTRACT FROM AUTHOR]

At present, the quality of material life and richness of the Chinese residents have been improved. Some people have high requirements for quality of their personal lives and put forward the problem of health preservation. Honey, as a nutritious food, is deeply loved by people. There are a large number of trace elements in honey, such as VC, VA, VD, VB1, and VB2. The honey from the Chinese honeybee has a very high nutritional value and plays an important role in the pollination and reproduction of some plants. Therefore, the Chinese honeybee plays a very significant role in the ecological environment. Moreover, it is protected as the main species resource of the country, which also fully proves the importance of the Chinese honeybee. Chinese bees can survive in various ecological and geographical environments in China and have strong heat as well as cold resistance. They can survive in the hot environment in the south and withstand the dynamics of severe cold in the north. At the same time, they can make full use of a small number of honey sources and have strong resistance to a variety of diseases and pests. In fact, there will be a variety of insect invasion problems in the beehive culture of Chinese bees, and it is necessary to accurately detect various diseases and pests during the breeding of Chinese bees. However, there are a large amount of insect invasion and various disease sources in the breeding stage of the Chinese bees. Therefore, in this paper, we use a deep learning algorithm to detect the insect invasion of the Chinese beehive culture and analyze the bee colonies in six bee farms in the province of Sichuan. In addition, we measure the common insect and disease indexes of the Chinese bee and analyze the parasitism rate, microsporidia infection rate, virus infection rate, and virus infection titer of bee colonies in overwintering and spring breeding. The experimental results show that the anti-insect invasion situation of bees in the six bee farms is significantly different; however, the antimite ability is basically the same. [ABSTRACT FROM AUTHOR]

Microsporidia are obligate intracellular parasites able to infest specifically a large range of species, including insects. The knowledge about the biology of microsporidial infections remains confined to mostly descriptive studies, including molecular approaches such as transcriptomics or proteomics. Thus, functional data to understand insect host defenses are currently lacking. Here, we have undertaken a genetic analysis of known host defenses of the Drosophila melanogaster using an infection model whereby Tubulinosema ratisbonensis spores are directly injected in this insect. We find that phagocytosis does confer some protection in this infection model. In contrast, the systemic immune response, extracellular reactive oxygen species, thioester proteins, xenophagy, and intracellular antiviral response pathways do not appear to be involved in the resistance against this parasite. Unexpectedly, several genes such as PGRP-LE seem to promote this infection. The prophenol oxidases that mediate melanization have different functions; PPO1 presents a phenotype similar to that of PGRP-LE whereas that of PPO2 suggests a function in the resilience to infection. Similarly, eiger and Unpaired3 , which encode two cytokines secreted by hemocytes display a resilience phenotype with a strong susceptibility to T. ratisbonensis. [ABSTRACT FROM AUTHOR]

The hygienic behavior in honey bees is a complex polygenic trait that serves as a natural defense mechanism against bacterial and fungal brood diseases and Varroa destructor mites infesting brood cells. The aim of this study was to evaluate the effect of a dietary amino acids and vitamins supplement "BEEWELL AminoPlus" on hygienic behavior of Apis mellifera colonies combating microsporidial and viral infections. The experiment was performed during a one-year period on 40 colonies alloted to five groups: one supplemented and infected with Nosema ceranae and four viruses (Deformed wing virus - DWV, Acute bee paralysis virus - ABPV, Chronic bee paralysis virus - CBPV and Sacbrood virus - SBV), three not supplemented, but infected with N. ceranae and/or viruses, and one negative control group. Beside the listed pathogens, honey bee trypanosomatids were also monitored in all groups. The supplement "BEEWELL AminoPlus" induced a significant and consistent increase of the hygienic behavior in spite of the negative effects of N. ceranae and viral infections. N. ceranae and viruses significantly and consistently decreased hygienic behavior, but also threatened the survival of bee colonies. The tested supplement showed anti-Nosema effect, since the N. ceranae infection level significantly and consistently declined only in the supplemented group. Among infected groups, only the supplemented one remained Lotmaria passim-free throughout the study. In conclusion, diet supplementation enhances hygienic behavior of honey bee colonies and helps them fight the most common infections of honey bees. [ABSTRACT FROM AUTHOR]