-
Parasites & Vectors Apr 2024In the context of climate change, a growing concern is that vector-pathogen or host-parasite interactions may be correlated with climatic factors, especially increasing...
BACKGROUND
In the context of climate change, a growing concern is that vector-pathogen or host-parasite interactions may be correlated with climatic factors, especially increasing temperatures. In the present study, we used a mosquito-microsporidian model to determine the impact of environmental factors such as temperature, humidity, wind and rainfall on the occurrence rates of opportunistic obligate microparasites (Microsporidia) in hosts from a family that includes important disease vectors (Culicidae).
METHODS
In our study, 3000 adult mosquitoes collected from the field over 3 years were analysed. Mosquitoes and microsporidia were identified using PCR and sequencing of the hypervariable V5 region of the small subunit ribosomal RNA gene and a shortened fragment of the cytochrome c oxidase subunit I gene, respectively.
RESULTS
DNA metabarcoding was used to identify nine mosquito species, all of which were hosts of 12 microsporidian species. The prevalence of microsporidian DNA across all mosquito samples was 34.6%. Microsporidian prevalence in mosquitoes was more frequent during warm months (> 19 °C; humidity < 65%), as was the co-occurrence of two or three microsporidian species in a single host individual. During warm months, microsporidian occurrence was noted 1.6-fold more often than during the cold periods. Among the microsporidians found in the mosquitoes, five (representing the genera Enterocytospora, Vairimorpha and Microsporidium) were positively correlated with an increase in temperature, whereas one (Hazardia sp.) was significantly correlated with a decrease in temperature. Threefold more microsporidian co-occurrences were recorded in the warm months than in the cold months.
CONCLUSIONS
These results suggest that the susceptibility of mosquitoes to parasite occurrence is primarily determined by environmental conditions, such as, for example, temperatures > 19 °C and humidity not exceeding 62%. Collectively, our data provide a better understanding of the effects of the environment on microsporidian-mosquito interactions.
Topics: Animals; Culicidae; Temperature; Humidity; Mosquito Vectors; Microsporidia; DNA
PubMed: 38605410
DOI: 10.1186/s13071-024-06254-0 -
Journal of Microbiology and... May 2024The increasing economic losses associated with growth retardation caused by (EHP), a microsporidian parasite infecting penaeid shrimp, require effective monitoring. The...
The Use of the Internal Transcribed Spacer Region for Phylogenetic Analysis of the Microsporidian Parasite Infecting Whiteleg Shrimp () and for the Development of a Nested PCR as Its Diagnostic Tool.
The increasing economic losses associated with growth retardation caused by (EHP), a microsporidian parasite infecting penaeid shrimp, require effective monitoring. The internal transcribed spacer (ITS)-1 region, the non-coding region of ribosomal clusters between 18S and 5.8S rRNA genes, is widely used in phylogenetic studies due to its high variability. In this study, the ITS-1 region sequence (~600-bp) of EHP was first identified, and primers for a polymerase chain reaction (PCR) assay targeting that sequence were designed. A newly developed nested-PCR method successfully detected the EHP in various shrimp ( and ) and related samples, including water and feces collected from Indonesia, Thailand, South Korea, India, and Malaysia. The primers did not cross-react with other hosts and pathogens, and this PCR assay is more sensitive than existing PCR detection methods targeting the small subunit ribosomal RNA (SSU rRNA) and spore wall protein (SWP) genes. Phylogenetic analysis based on the ITS-1 sequences indicated that the Indonesian strain was distinct (86.2% nucleotide sequence identity) from other strains collected from Thailand and South Korea, and also showed the internal diversity among Thailand ( = 7, divided into four branches) and South Korean ( = 5, divided into two branches) samples. The results revealed the ability of the ITS-1 region to determine the genetic diversity of EHP from different geographical origins.
Topics: Enterocytozoon; Penaeidae; Animals; Phylogeny; DNA, Ribosomal Spacer; Polymerase Chain Reaction; Microsporidiosis; DNA, Fungal; DNA Primers; Feces; Sequence Analysis, DNA; Thailand
PubMed: 38563108
DOI: 10.4014/jmb.2401.01010 -
BMC Genomics Apr 2024Congenital infection caused by vertical transmission of microsporidia N. bombycis can result in severe economic losses in the silkworm-rearing industry....
Congenital infection caused by vertical transmission of microsporidia N. bombycis can result in severe economic losses in the silkworm-rearing industry. Whole-transcriptome analyses have revealed non-coding RNAs and their regulatory networks in N. bombycis infected embryos and larvae. However, transcriptomic changes in the microsporidia proliferation and host responses in congenitally infected embryos and larvae remains unclear. Here, we simultaneously compared the transcriptomes of N. bombycis and its host B. mori embryos of 5-day and larvae of 1-, 5- and 10-day during congenital infection. For the transcriptome of N. bombycis, a comparison of parasite expression patterns between congenital-infected embryos and larva showed most genes related to parasite central carbon metabolism were down-regulated in larvae during infection, whereas the majority of genes involved in parasite proliferation and growth were up-regulated. Interestingly, a large number of distinct or shared differentially expressed genes (DEGs) were revealed by the Venn diagram and heat map, many of them were connected to infection related factors such as Ricin B lectin, spore wall protein, polar tube protein, and polysaccharide deacetylase. For the transcriptome of B. mori infected with N. bombycis, beyond numerous DEGs related to DNA replication and repair, mRNA surveillance pathway, RNA transport, protein biosynthesis, and proteolysis, with the progression of infection, a large number of DEGs related to immune and infection pathways, including phagocytosis, apoptosis, TNF, Toll-like receptor, NF-kappa B, Fc epsilon RI, and some diseases, were successively identified. In contrast, most genes associated with the insulin signaling pathway, 2-oxacarboxylic acid metabolism, amino acid biosynthesis, and lipid metabolisms were up-regulated in larvae compared to those in embryos. Furthermore, dozens of distinct and three shared DEGs that were involved in the epigenetic regulations, such as polycomb, histone-lysine-specific demethylases, and histone-lysine-N-methyltransferases, were identified via the Venn diagram and heat maps. Notably, many DEGs of host and parasite associated with lipid-related metabolisms were verified by RT-qPCR. Taken together, simultaneous transcriptomic analyses of both host and parasite genes lead to a better understanding of changes in the microsporidia proliferation and host responses in embryos and larvae in N. bombycis congenital infection.
Topics: Animals; Transcriptome; Larva; Histones; Lysine; Nosema; Gene Expression Profiling; Cell Proliferation; Lipids; Bombyx
PubMed: 38556880
DOI: 10.1186/s12864-024-10236-y -
Microorganisms Mar 2024is an obligate intracellular microsporidium, which poses a significant threat to ayu (). In vitro cultivation models are invaluable tools for investigating...
is an obligate intracellular microsporidium, which poses a significant threat to ayu (). In vitro cultivation models are invaluable tools for investigating intracellular microorganisms, including . In this study, we attempted to in vitro cultivate using primary cultures derived from ayu monocytes/macrophages (MO/MΦ), a murine-derived macrophage cell line RAW264.7, and the epithelioma papulosum cyprini (EPC) cell line. The results demonstrated that MO/MΦ infected with spores exhibited a pronounced immune response which was presented by rapidly high expression levels of inflammatory cytokines, such as , , , and , and detached within 96 h post-infection (hpi). Infected RAW264.7 cells remained capable of stable passage yet exhibited cellular deformation with a decrease in intracellular spores occurring around 8 days post-infection (dpi). In contrast, EPC cells promised a substantial parasite population, and the cytokine expression levels returned to normal by 8 dpi. In addition, spores recovered from EPC cells could infect young ayu, suggesting that EPC cells might be used as an in vitro cultivation system for .
PubMed: 38543573
DOI: 10.3390/microorganisms12030522 -
Insects Feb 2024Discoveries of endemic species highlight areas of biogeographic and conservation interest. Endemic species, however, are often morphologically disguised as more common...
Discoveries of endemic species highlight areas of biogeographic and conservation interest. Endemic species, however, are often morphologically disguised as more common and widespread species. The larval polytene chromosomes revealed a new species of black fly, , from the Djurdjura Mountains of northern Algeria, and its female, male, pupa, and larva are described. The species is chromosomally unique; none of its 11 chromosomal rearrangements are shared with other species. Although the new species structurally resembles (Meigen) with which it previously has been confused, it can be distinguished from all other known species of in the Western Palearctic based on at least one character in each described life stage. Symbiotic organisms included two species of microsporidia, at least one of which is probably undescribed, one unknown protozoan pathogen novel in simuliids, and the trichomycete fungus Léger and Duboscq. Associated simuliid species included at least one new species of the genus . The new species of is tentatively considered endemic to the mountains of northern Algeria but might be expected in the mountains of eastern Morocco and northern Tunisia and perhaps in Sicily. If its endemic status holds, it would be the only nominal species of black fly unique to Algeria.
PubMed: 38535346
DOI: 10.3390/insects15030150 -
Journal of Fungi (Basel, Switzerland) Mar 2024Chaperonin containing tailless complex polypeptide 1 (CCT) is a molecular chaperone protein that consists of eight completely different subunits and assists in the...
Chaperonin containing tailless complex polypeptide 1 (CCT) is a molecular chaperone protein that consists of eight completely different subunits and assists in the folding of newly synthesized peptides. The zeta subunit of CCT is a regulatory factor for the folding and assembly of cytoskeletal proteins as individuals or complexes. In this study, the zeta subunit of (NbCCTζ) is identified for the first time. The complete ORF of the gene is 1533 bp in length and encodes a 510 amino acid polypeptide. IFA results indicate that NbCCTζ is colocalized with actin and β-tubulin in the cytoplasm during the proliferative phase and that NbCCTζ is completely colocalized with NbCCTα in the cytoplasm of throughout the entire life cycle. Furthermore, the yeast two-hybrid assay revealed that the NbCCTζ interacts with NbCCTα. The transcriptional level of is significantly downregulated by knocking down the gene, while the transcriptional level of is downregulated after knocking down the gene. These results suggest that NbCCTζ may play a vital role in the proliferation of by coordinating with NbCCTα.
PubMed: 38535237
DOI: 10.3390/jof10030229 -
Food and Waterborne Parasitology Jun 2024is one of the most prevalent microsporidia species, responsible for more than 90% of human and animal microsporidiosis. Microsporidia species, particularly are...
is one of the most prevalent microsporidia species, responsible for more than 90% of human and animal microsporidiosis. Microsporidia species, particularly are frequently reported from waterborne and foodborne outbreaks. Therefore, early detection is crucial in clinics and outbreak investigations. This study aimed to design a loop-mediated isothermal amplification (LAMP) for rapid detection of Total DNA was extracted from 30 -positive samples, which had been confirmed with nested PCR. LAMP primers were designed based on the identical fragment of small subunit ribosomal RNA () gene. LAMP reactions were performed at 63 °C for 60 min. The sensitivity and specificity of the assay were analyzed and the results of amplification were compared to real-time PCR. Our results showed that the LAMP assay successfully amplified 25/30 (83.3%) samples. The specificity results indicated no false positive with other microorganisms. Furthermore, the LAMP method exhibited a sensitivity (limit of detection, LoD) as low as 34 ag/μL of total DNA. Compared to the LAMP assay, real-time PCR was able to detect all 30 nested PCR-positive samples. Our findings showed that the LAMP assay was able to detect 83.3% of positive samples. Although the current assay was not able to detect all nested PCR-positive samples, the lack of need for specific instruments, rapid processes, and high specificity makes LAMP assay a suitable tool for screening.
PubMed: 38523772
DOI: 10.1016/j.fawpar.2024.e00225 -
Microbiology Resource Announcements Apr 2024We report the draft whole-genome assembly of sp. MB a symbiotic malaria-transmission-blocking microsporidian isolated from in Kenya. The whole-genome sequence of sp....
We report the draft whole-genome assembly of sp. MB a symbiotic malaria-transmission-blocking microsporidian isolated from in Kenya. The whole-genome sequence of sp. MB has a length of 5,908,979 bp, 2,335 contigs, and an average GC content of 31.12%.
PubMed: 38509052
DOI: 10.1128/MRA.00903-23 -
Malaria Journal Mar 2024Recently, bacterial endosymbiont, including Wolbachia and Microsporidia were found to limit the infection of Anopheles mosquitoes with Plasmodium falciparum. This study...
BACKGROUND
Recently, bacterial endosymbiont, including Wolbachia and Microsporidia were found to limit the infection of Anopheles mosquitoes with Plasmodium falciparum. This study aimed to investigate the natural presence of key transmission-blocking endosymbionts in Anopheles gambiae and Anopheles coluzzii in Southern Benin.
METHODS
The present study was conducted in seven communes (Cotonou, Porto-Novo, Aguégués, Ifangni, Pobè Athiémé, and Grand-Popo) of Southern Benin. Anopheles were collected using indoor/outdoor Human Landing Catches (HLCs) and Pyrethrum Spray Catches (PSCs). Following morphological identification, PCR was used to identify An. gambiae sensu lato (s.l.) to species level and to screen for the presence of both Wolbachia and Microsporidia. Plasmodium falciparum sporozoite infection was also assessed using ELISA.
RESULTS
Overall, species composition in An. gambiae s.l. was 53.7% An. coluzzii, while the remainder was An. gambiae sensu stricto (s.s.). Combined data of the two sampling techniques revealed a mean infection prevalence with Wolbachia of 5.1% (95% CI 0.90-18.6) and 1.3% (95% CI 0.07-7.8) in An. gambiae s.s. and An. coluzzii, respectively. The mean infection prevalence with Microsporidia was 41.0% (95% CI 25.9-57.8) for An. gambiae s.s. and 57.0% (95% CI 45.4-67.9) for An. coluzzii. Wolbachia was only observed in Ifangni, Pobè, and Cotonou, while Microsporidia was detected in all study communes. Aggregated data for HLCs and PSCs showed a sporozoite rate (SR) of 0.80% (95% CI 0.09-2.87) and 0.69% (95% CI 0.09-2.87) for An. gambiae and An. coluzzii, respectively, with a mean of 0.74% (95% CI 0.20-1.90). Of the four individual mosquitoes which harboured P. falciparum, none were also infected with Wolbachia and one contained Microsporidia.
CONCLUSIONS
The present study is the first report of natural infections of field-collected An. gambiae s.l. populations from Benin with Wolbachia and Microsporidia. Sustained efforts should be made to widen the spectrum of bacteria identified in mosquitoes, with the potential to develop endosymbiont-based control tools; such interventions could be the game-changer in the control of malaria and arboviral disease transmission.
Topics: Animals; Humans; Benin; Anopheles; Wolbachia; Cross-Sectional Studies; Mosquito Vectors; Malaria, Falciparum; Pyrethrins; Sporozoites
PubMed: 38468292
DOI: 10.1186/s12936-024-04906-1 -
PLoS Biology Mar 2024Protein quality control pathways play important roles in resistance against pathogen infection. For example, the conserved transcription factor SKN-1/NRF up-regulates...
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.
Topics: Animals; Caenorhabditis elegans; Transcription Factors; DNA-Binding Proteins; Proteasome Endopeptidase Complex; Caenorhabditis elegans Proteins; Immunity, Innate; Bacterial Infections
PubMed: 38466732
DOI: 10.1371/journal.pbio.3002543