Microsporidian Parasite Research Articles

Establishment of a TaqMan probe-based qPCR assay for detecting microsporidia Enterospora epinepheli in grouper.

Enterospora epinepheli is an intranuclear microsporidian parasite causing serious emaciative disease in hatchery-bred juvenile groupers (Epinephelus spp.). Rapid and sensitive detection is urgently needed as its chronic infection tends to cause emaciation as well as white faeces syndrome and results in fry mortality. This study established a TaqMan probe-based real-time quantitative PCR assays targeting the small subunit rRNA (SSU) gene of E. epinepheli. The relationship between the standard curve of cycle threshold (Ct) and the logarithmic starting quantity (SQ) was determined as Ct = -3.177 lg (SQ) + 38.397. The correlation coefficient (R2 ) was 0.999, and the amplification efficiency was 106.4%. The detection limit of the TaqMan probe-based qPCR assay was 1.0 × 101 copies/μL and that is 100 times sensitive than the traditional PCR method. There is no cross-reaction with other aquatic microsporidia such as Ecytonucleospora hepatopenaei, Nucleospora hippocampi, Potaspora sp., Ameson portunus. The intra-assay and inter-assay showed great repeatability and reproducibility. In addition, the test of clinical samples showed that this assay effectively detected E. epinepheli in the grouper's intestine tissue. The established TaqMan qPCR assays will be a valuable diagnostic tool for the epidemiological investigation as well as prevention and control of E. epinepheli.

Dynamic Interplay of Metabolic and Transcriptional Responses in Shrimp during Early and Late Infection Stages of Enterocytozoon hepatopenaei (EHP)

Enterocytozoon hepatopenaei (EHP) is a microsporidian parasite that infects Litopenaeus vannamei, causing severe hepatopancreatic microsporidiosis (HPM) and resulting in significant economic losses. This study utilizes a combined analysis of transcriptomics and metabolomics to unveil the dynamic molecular interactions between EHP and its host, the Pacific white shrimp, during the early and late stages of infection. The results indicate distinct immunological, detoxification, and antioxidant responses in the early and late infection phases. During early EHP infection in shrimp, immune activation coincides with suppression of genes like Ftz-F1 and SEPs, potentially aiding parasitic evasion. In contrast, late infection shows a refined immune response with phagocytosis-enhancing down-regulation of Ftz-F1 and a resurgence in SEP expression. This phase is characterized by an up-regulated detoxification and antioxidant response, likely a defense against the accumulated effects of EHP, facilitating a stable host–pathogen relationship. In the later stages of infection, most immune responses return to baseline levels, while some immune genes remain active. The glutathione antioxidant system is suppressed early on but becomes activated in the later stages. This phenomenon could facilitate the early invasion of EHP while assisting the host in mitigating oxidative damage caused by late-stage infection. Notably, there are distinctive events in polyamine metabolism. Sustained up-regulation of spermidine synthase and concurrent reduction in spermine levels suggest a potential role of polyamines in EHP development. Throughout the infection process, significant differences in genes such as ATP synthase and hexokinase highlight the continuous influence on energy metabolism pathways. Additionally, growth-related pathways involving amino acids such as tryptophan, histidine, and taurine are disrupted early on, potentially contributing to the growth inhibition observed during the initial stages of infection. In summary, these findings elucidate the dynamic interplay between the host, Litopenaeus vannamei, and the parasite, EHP, during infection. Specific phase differences in immune responses, energy metabolism, and antioxidant processes underscore the intricate relationship between the host and the parasite. The disruption of polyamine metabolism offers a novel perspective in understanding the proliferation mechanisms of EHP. These discoveries significantly advance our comprehension of the pathogenic mechanisms of EHP and its interactions with the host.

Prevalence and Genetic Diversity of a Microsporidian Parasite in the Black Imported Fire Ant and Its Social Parasitic Ant (Formicidae: Myrmicinae: Solenopsis) in Buenos Aires Province, Argentina.

Microsporidia are natural pathogens of arthropods and have been used as biological control against insect pests. In the United States, efforts to control the invasive Red Imported Fire Ant, Solenopsis invicta, and Black Imported Fire Ant, Solenopsis richteri, have included the use of the microsporidium, Kneallhazia solenopsae. However, there is limited information about the genetic differences among the microsporidian variants found in S. invicta and in S. richteri. In this study, we assessed the prevalence and genetic diversity of K. solenopsae in native populations of S. richteri in Argentina (South America). Additionally, we examined the social parasitic ant, Solenopsis daguerrei, which is found in some S. richteri nests, for the presence of this microsporidium. The survey of 219 S. richteri nests revealed K. solenopsae infections in all five sites analyzed, with 28 colonies (12.8%) positive for the microsporidium. Among the 180 S. daguerrei individuals collected, seven ants (3.9%) from three sites tested positive for K. solenopsae. Phylogenetic analyses of the microsporidian variants present in S. richteri and S. daguerrei based on partial small subunit ribosomal gene sequences (SSU rRNA) showed that both ant species shared the same variant, which is different from the ones found in S. invicta. Further studies are needed to determine the pathogenicity of genetically different K. solenopsae variants among Solenopsis species.

Increased virulence due to multiple infection in Daphnia leads to limited growth in 1 of 2 co-infecting microsporidian parasites.

Recent outbreaks of various infectious diseases have highlighted the ever-present need to understand the drivers of the outbreak and spread of disease. Although much of the research investigating diseases focuses on single infections, natural systems are dominated by multiple infections. These infections may occur simultaneously, but are often acquired sequentially, which may alter the outcome of infection. Using waterfleas (Daphnia magna) as a model organism, we examined the outcome of sequential and simultaneous multiple infections with 2 microsporidian parasites (Ordospora colligata and Hamiltosporidium tvaerminnensis) in a fully factorial design with 9 treatments and 30 replicates. We found no differences between simultaneous and sequential infections. However, H. tvaerminnensis fitness was impeded by multiple infection due to increased host mortality, which gave H. tvaerminnensis less time to grow. Host fecundity was also reduced across all treatments, but animals infected with O. colligata at a younger age produced the fewest offspring. As H. tvaerminnensis is both horizontally and vertically transmitted, this reduction in offspring may have further reduced H. tvaerminnensis fitness in co-infected treatments. Our findings suggest that in natural populations where both species co-occur, H. tvaerminnensis may evolve to higher levels of virulence following frequent co-infection by O. colligata.

Diseases and Pests Harmful to Honeybees (Apis spp.) and Their Management Tactics: A Review

Honeybees (Apis spp.) play a vital role in pollination and contribute significantly to global agriculture. However, the health and survival of honeybee colonies are threatened by a range of harmful insect pests and diseases. This review provides an overview of the major pests and diseases that impact honeybee colonies and explores the diverse management tactics employed to mitigate their effects. The primary focus is on key pests such as the Varroa mite (Varroa destructor), which inflicts considerable damage by parasitizing adult bees and their brood, facilitating the transmission of debilitating viruses. Management strategies for Varroa mites encompass chemical treatments, integrated pest management (IPM) approaches, and organic treatments to curtail mite populations and prevent colony collapse. Another significant threat discussed is the small hive beetle (Aethina tumida), a beetle species that disrupts hives by laying eggs in comb and consuming stored resources. Hive manipulation, traps, and chemical controls are among the tactics utilized to control small hive beetle populations and mitigate their impact. The review also addresses wax moths (Galleria mellonella and Achroia grisella), which target hive comb, potentially causing comb destruction and honey fermentation. Hive maintenance, freezing, and pheromone traps are outlined as effective strategies to combat wax moth infestations. Additionally, the review covers Nosema infections (Nosema apis and Nosema ceranae), and microsporidian parasites that compromise adult bee health by weakening immune systems and reducing foraging efficiency. Hive hygiene, medications, and genetic selection are explored as methods to manage Nosema infections and bolster colony resilience. To effectively manage these harmful pests and diseases, a holistic and integrated approach is recommended. Beekeepers are encouraged to remain informed about the latest research findings and implement appropriate measures, considering local conditions and sustainable beekeeping practices. By combining scientific knowledge with practical application, the resilience of honeybee colonies can be enhanced, ensuring the vital role they play in pollination and global food security.

Metagenomic Studies Reveal the Evidence of Akkermansia muciniphila and Other Probiotic Bacteria in the Gut of Healthy and Enterocytozoon hepatopenaei (EHP)-Infected Farmed Penaeus vannamei.

Penaeus vannamei (whiteleg shrimp) is the most widely cultured shrimp globally. Enterocytozoon hepatopenaei (EHP), a microsporidian parasite, infects P. vannamei and causes severe growth retardation, subsequent production, and economic losses in the shrimp culture. The influence of EHP infection in the shrimp gut microbiota is poorly studied, and this would be an interesting area to investigate since the gut microbiome of shrimp influences a number of key host processes such as digestion and immunity. In this study, a metagenomic approach was followed to compare the overall species richness of the gut microbiota of EHP-infected and healthy P. vannamei. Bacterial genomic DNA from the healthy and EHP-infected gut sample were profiled for the bacterial 16S rRNA gene, targeting the V3-V4 conserved region. Operational taxonomic units (OTUs), an approximation of definitive taxonomic identity, were identified based on the sequence similarity within the sample reads and clustered together using a cut-off of 97% identity using UCLUST. The OTUs were then used for the computation of alpha diversity and beta diversity for each sample. EHP-infected gut sample showed lower bacterial abundance throughout the family, class, order, genus, and species levels when compared to healthy gut sample. This study shows that the shrimp gut microbiota is sensitive and exhibits a high level of plasticity during a microsporidian infection like EHP. Furthermore, Akkermansia muciniphila, a novel probiotic bacterium, has been reported in the shrimp gut for the first time.

CryoEM reveals that ribosomes in microsporidian spores are locked in a dimeric hibernating state.

Translational control is an essential process for the cell to adapt to varying physiological or environmental conditions. To survive adverse conditions such as low nutrient levels, translation can be shut down almost entirely by inhibiting ribosomal function. Here we investigated eukaryotic hibernating ribosomes from the microsporidian parasite Spraguea lophii in situ by a combination of electron cryo-tomography and single-particle electron cryo-microscopy. We show that microsporidian spores contain hibernating ribosomes that are locked in a dimeric (100S) state, which is formed by a unique dimerization mechanism involving the beak region. The ribosomes within the dimer are fully assembled, suggesting that they are ready to be activated once the host cell is invaded. This study provides structural evidence for dimerization acting as a mechanism for ribosomal hibernation in microsporidia, and therefore demonstrates that eukaryotes utilize this mechanism in translational control.

Molecular characterization of turtle-like protein in whiteleg shrimp (Litopenaeus vannamei) and its role in Enterocytozoon hepatopenaei infection.

Enterocytozoon hepatopenaei (EHP) is a microsporidian parasite that infects shrimp hepatopancreas, causing growth retardation and disease susceptibility. Knowledge of the host-pathogen molecular mechanisms is essential to understanding the microsporidian pathogenesis. Turtle-like protein (TLP) is part of the immunoglobulin superfamily of proteins, which is widely distributed in the animal kingdom. TLP has multiple functions, such as cell surface receptors and cell adhesion molecules. The spore wall proteins (SWPs) of microsporidia are involved in the infection mechanisms. Some SWPs are responsible for spore adherence, which is part of the activation and host cell invasion processes. Previous studies showed that TLP from silkworms (Bombyx mori) interacted with SWP26, contributing to the infectivity of Nosema bombycis to its host. In this study, we identified and characterized for the first time, the Litopenaeus vannamei TLP gene (LvTLP), which encodes an 827-aa protein (92.4 kDa) composed of five immunoglobulin domains, two fibronectin type III domains, and a transmembrane region. The LvTLP transcript was expressed in all tested tissues and upregulated in the hepatopancreas at 1 and 7 days post-cohabitation (dpc) and at 9 dpc in hemocytes. To identify the LvTLP binding counterpart, recombinant (r)LvTLP and recombinant spore wall protein 1 of (rEhSWP1) were produced in Escherichia coli. Coimmunoprecipitation and enzyme-linked immunosorbent assays demonstrated that rLvTLP interacted with rEhSWP with high affinity (KD = 1.20 × 10-7 M). In EHP-infected hepatopancreases, LvTLP was clustered and co-localized with some of the developing EHP plasmodia. Furthermore, LvTLP gene silencing reduced the EHP copy numbers compared with those of the control group, suggesting the critical role of LvTLP in EHP infection. These results provide insight into the molecular mechanisms of the host-pathogen interactions during EHP infection.

Molecular characterization of a microsporidian parasite, Ovipleistophora ovariae SW02 infecting the snakehead fish, Channa punctatus from the river Gomti, Lucknow, India

Molecular characterization of a microsporidian parasite, Ovipleistophora ovariae SW02 infecting the snakehead fish, Channa punctatus from the river Gomti, Lucknow, India

Release of live baitfish by recreational anglers drives fish pathogen introduction risk.

Emerging diseases of wildlife are an existential threat to biodiversity, and human-mediated movements of live animals are a primary vector of their spread. Wildlife disease risk analyses offer an appealing alternative to precautionary approaches because they allow for explicit quantification of uncertainties and consideration of tradeoffs. Such considerations become particularly important in high-frequency invasion pathways with hundreds of thousands of individual vectors, where even low pathogen prevalence can lead to substantial risk. The purpose of this study was to examine the landscape-level dynamics of human behavior-mediated pathogen introduction risk in the context of a high-frequency invasion pathway. One such pathway is the use and release of live fish used as bait by recreational anglers. We used a stochastic risk assessment model parameterized by angler survey data from Minnesota, USA, to simulate one year of fishing in Minnesota and estimate the total number of risky trips for each of three pathogens: viral hemorrhagic septicemia virus, the microsporidian parasite Ovipleistophora ovariae, and the Asian fish tapeworm Schizocotyle acheilognathi. We assessed the number of introductionsunder four scenarios: current/baseline conditions, outbreak conditions (increased pathogen prevalence), source-focused control measures (decreased pathogen prevalence), and angler-focused control measures (decreased rates of release). We found that hundreds of thousands of introduction events can occur per year, even for regulated pathogens at low pathogen prevalence. Reducing the rate of illegal baitfish release had significant impact on risky trips in scenarios where a high number of anglers were involved, but was less impactful in circumstances with limited outbreaks and fewer affected anglers. In contrast, reducing pathogen prevalence in the source populations of baitfish had relatively little impact. In order to make meaningful changes in pathogen introduction risk, managers should focus efforts on containing local outbreaks and reducing illegal baitfish release to reduce pathogen introduction risk. Our study also demonstrates the risk associated with high-frequency invasion pathways and the importance of incorporating human behaviors into wildlife disease models and risk assessments.

Mysterious microsporidians: springtime outbreaks of disease in Daphnia communities in shallow pond ecosystems.

Parasites can play key roles in ecosystems, especially when they infect common hosts that play important ecological roles. Daphnia are critical grazers in many lentic freshwater ecosystems and typically reach peak densities in early spring. Daphnia have also become prominent model host organisms for the field of disease ecology, although most well-studied parasites infect them in summer or fall. Here, we report field patterns of virulent microsporidian parasites that consistently infect Daphnia in springtime, in a set of seven shallow ponds in Georgia, USA, sampled every 3-4weeks for 18months. We detected two distinct parasite taxa, closely matching sequences of Pseudoberwaldia daphniae and Conglomerata obtusa, both infecting all three resident species of Daphnia: D. ambigua, D. laevis, and D. parvula. To our knowledge, neither parasite has been previously reported in any of these host species or anywhere in North America. Infection prevalence peaked consistently in February-May, but the severity of these outbreaks differed substantially among ponds. Moreover, host species differed markedly in terms of their maximum infection prevalence (5% [D. parvula] to 72% [D. laevis]), mean reduction of fecundity when infected (70.6% [D. ambigua] to 99.8% [D. laevis]), mean spore yield (62,000 [D. parvula] to 377,000 [D. laevis] per host), and likelihood of being infected by each parasite. The timing and severity of the outbreaks suggests that these parasites could be impactful members of these shallow freshwater ecosystems, and that the strength of their effects is likely to hinge on the composition of ponds' zooplankton communities.

Engineered gut symbiont inhibits microsporidian parasite and improves honey bee survival

Honey bees (Apis mellifera) are critical agricultural pollinators as well as model organisms for research on development, behavior, memory, and learning. The parasite Nosema ceranae, a common cause of honey bee colony collapse, has developed resistance to small-molecule therapeutics. An alternative long-term strategy to combat Nosema infection is therefore urgently needed, with synthetic biology offering a potential solution. Honey bees harbor specialized bacterial gut symbionts that are transmitted within hives. Previously, these have been engineered to inhibit ectoparasitic mites by expressing double-stranded RNA (dsRNA) targeting essential mite genes, via activation of the mite RNA interference (RNAi) pathway. In this study, we engineered a honey bee gut symbiont to express dsRNA targeting essential genes of N. ceranae via the parasite's own RNAi machinery. The engineered symbiont sharply reduced Nosema proliferation and improved bee survival following the parasite challenge. This protection was observed in both newly emerged and older forager bees. Furthermore, engineered symbionts were transmitted among cohoused bees, suggesting that introducing engineered symbionts to hives could result in colony-level protection.

Possible seasonal and diurnal modulation of Gammarus pulex (Crustacea, Amphipoda) drift by microsporidian parasites

In lotic freshwater ecosystems, the drift or downstream movement of animals (e.g., macroinvertebrates) constitutes a key dispersal pathway, thus shaping ecological and evolutionary patterns. There is evidence that macroinvertebrate drift may be modulated by parasites. However, most studies on parasite modulation of host drifting behavior have focused on acanthocephalans, whereas other parasites, such as microsporidians, have been largely neglected. This study provides new insight into possible seasonal and diurnal modulation of amphipod (Crustacea: Gammaridae) drift by microsporidian parasites. Three 72 h drift experiments were deployed in a German lowland stream in October 2021, April, and July 2022. The prevalence and composition of ten microsporidian parasites in Gammarus pulex clade E varied seasonally, diurnally, and between drifting and stationary specimens of G. pulex. Prevalence was generally higher in drifting amphipods than in stationary ones, mainly due to differences in host size. However, for two parasites, the prevalence in drift samples was highest during daytime suggesting changes in host phototaxis likely related to the parasite’s mode of transmission and site of infection. Alterations in drifting behavior may have important implications for G. pulex population dynamics and microsporidians’ dispersal. The underlying mechanisms are more complex than previously thought.

Potential of Fumagillin Against <i>Nosema</i> SP., a Microsporidian Endoparasite of Honey Bee <i>Apis mellifera</i> L

Nosema apis and Nosema ceranae are two kinds of microsporidian parasites that cause the recently identified illness known as nosemosis in European honey bees (Apis mellifera). 10 samples of adult honey bees from each infected colony of a single apiary were collected, located in the village of Handia, district of Prayagraj, India, from September 2022 to January 2023 to investigate the frequency of intestinal infections and the effectiveness of fumagillin antibiotic against nosemosis in A. mellifera with the use of monocular LED optical microscopy and a haemocytometer, the mean and median spore count of each composite sample was calculated. It was observed that 5% of the infected gut samples contained Nosema spores. The results show that 10% of colonies tested positive for Nosema, and fumagillin is effective against nosemosis.

Symptomatic Infection with Vairimorpha spp. Decreases Diapause Survival in a Wild Bumble Bee Species (Bombus griseocollis).

Vairimorpha, a microsporidian parasite (previously classified as Nosema), has been implicated in the decline of wild bumble bee species in North America. Previous studies examining its influence on colony performance have displayed variable results, from extremely detrimental effects to no observable influence, and little is known about the effects it has on individuals during the winter diapause, a bottleneck for survival in many annual pollinators. Here, we examined the effect of Vairimorpha infection, body size, and mass on diapause survival in Bombus griseocollis gynes. We demonstrate that gyne survival length in diapause is negatively affected by symptomatic Vairimorpha infection of the maternal colony but does not correlate with individual pathogen load. Our findings further indicate that increased body mass offers a protective effect against mortality during diapause in infected, but not in healthy, gynes. This suggests that access to adequate nutritional resources prior to diapause might offset the harmful effect of Vairimorpha infection.

Pathology, epidemiology, and phylogeny of mussel egg disease due to the microsporidian Steinhausia mytilovum (Field, 1924) in the Mediterranean mussel (Mytilus galloprovincialis)

Microsporidia are well known fungal pathogens of aquatic animals. However, the taxonomy of microsporidia is generally poorly resolved, which has consequently constrained our understanding of their pathology and epidemiology in marine animals. To date, microsporidia have been reported in both bivalves and gastropods, and microsporidia from mollusks have been classified in different genera. Despite ongoing work to better describe these genera, including detailed microscopic and ultrastructural images, so far we lack information on microsporidian phylogeny and pathogenicity of species within these genera. Here we investigate the microsporidian parasite Steinhausia mytilovum associated with the mussel, Mytilus galloprovincialis, in natural beds and farms along coast of southern Italy. A survey of M. galloprovincialis was conducted in 13 mussel farms and one natural bed between 2009 and 2020. We found the presence of S. mytilovum in 10 of the investigated farms, with a prevalence ranging between 14 and 100% of female mussels, depending on the population and season in which they were sampled. The parasite developed in the oocytes within a sporophorous vesicle (SV) where it produced 1–3 spores per cell, both in the cytoplasm and in the nucleus. Stenhausia mytilovum elicited an infiltrative (24.8%) or a strong capsular inflammatory response (43.4%) at gonadal follicles and surrounding vesicular connective tissue, in some cases accompanied by gonadal atresia (24.8%), leading to loss of gonadal architecture. In 7% of cases no reaction was observed. Ultrastructural observations revealed a mitochondrial re-organization to interact with all the phases of parasite development; the mitochondria were arranged outside the parasitophorous vesicle (PV) or directly interacting with the spore inside vesicle. There are five taxonomic clades of microsporidians as identified by SSU ribosomal gene sequence data. Maximum likelihood analysis assigned S. mytilovum within the Clade IV, defined as the Class Terresporidia, with closest genetic relationship (83.6% identity) to an undetermined invertebrate ovarian microsporidian. The constant presence, prevalence, and severity of S. mytilovum in coastline populations of M. galloprovincialis populations in southern Italy may indirectly reflect immunocompetence at both individual and population levels.

New insights into the genome and transmission of the microsporidian pathogen Nosema muscidifuracis.

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. Here, we report the first assembly of the N. muscidifuracis genome (14,397,169 bp 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 Nosema-cured 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. 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.

Detection of Enterocytozoon hepatopenaei (EHP) (microsporidia) in several species of potential macrofauna-carriers from shrimp (Penaeus vannamei) ponds in Malaysia

Infection by the microsporidian parasite Enterocytozoon hepatopenaei (EHP) has become a significant problem in the shrimp cultivation industry in Asian countries like Thailand, China, India, Vietnam, Indonesia, and Malaysia. The outbreak of this microsporidian parasite is predominantly related to the existence of macrofauna-carriers of EHP. However, information about potential macrofauna-carriers of EHP in rearing ponds is still limited. In this study, the screening of EHP in potential macrofauna-carriers was conducted in farming ponds of Penaeus vannamei in three states in Malaysia, namely Penang, Kedah, and Johor. A total of 82 macrofauna specimens (phyla: Arthropoda, Mollusca, and Chordata) were amplified through a polymerase chain reaction (PCR) assay targeting genes encoding spore wall proteins (SWP) of EHP. The PCR results showed an average prevalence of EHP (82.93%) from three phyla (Arthropoda, Mollusca and Chordata). The phylogenetic tree generated from the macrofauna sequences was revealed to be identical to the EHP-infected shrimp specimens from Malaysia (MW000458, MW000459, and MW000460), as well as those from India (KY674537), Thailand (MG015710), Vietnam (KY593132), and Indonesia (KY593133). These findings suggest that certain macrofauna species in shrimp ponds of P. vannamei are carriers of EHP spores and could be potential transmission vectors. This study provides preliminary information for the prevention of EHP infections that can be initiated at the pond stage by eradicating macrofauna species identified as potential vectors.

Identification techniques to prevent the current emerging disease hepatopancreatic microsporidiosis in white shrimp Penaeus vannamei: an overview

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.

High diversity of microsporidian parasites and new planktonic hosts in freshwater and marine ecosystems

AbstractMicrosporidia are a large group of obligate intracellular eukaryotic parasites related to fungi primarily known as parasites of vertebrates and invertebrates. They are well described as parasites of organisms of interest (e.g., edible fish and crustaceans, honeybees, bioindicators such as daphnia, humans) on which they can have an important impact (e.g., reduced survival or fecundity and sex ratio distortion). However, their diversity in aquatic environments, especially in marine ecosystems, has been greatly understudied since they are not targeted by classical eukaryotic primers used in metabarcoding studies. Moreover, little is known about their hosts among protists or microzooplankton and therefore about their impact on the trophic food web functioning. In this work, we sampled 15 different sites across marine and freshwater environments, size‐fractioned the samples, and used microsporidian specific primers associated with metabarcoding to study the microsporidian diversity (and the associated spatial variation). Co‐occurrence networks as well as tyramide signal amplification‐fluorescent in situ hybridization were used to link potential hosts (planktonic eukaryotes < 150 μm) and Microsporidia diversity. Our analysis unraveled a large microsporidian diversity which was widely divergent between the two environments studied. In both of them, an important part of this diversity was not affiliated to a genus, suggesting an important reservoir of new microsporidian species and thus new hosts among planktonic eukaryotes. Co‐occurrence networks and fluorescence microscopy showed for the first‐time associations between Microsporidia and dinoflagellates in freshwater and marine environments.