Search Results (61)

Glycoside hydrolase family 1 (GH1) enzymes are essential for plant cell wall digestion and the detoxification of plant metabolites in insects, yet their evolutionary history in Lepidoptera remains unresolved. This study systematically identified GH1 genes across 61 Lepidopteran genomes and analyzed their evolutionary dynamics. In addition, the expression profiles of GH1 genes in the silkworm (Bombyx mori) across various developmental stages and tissues were related to their evolutionary histories. A total of 996 GH1 genes were annotated and classified into 11 groups, with each showing distinct species diversity. Gene duplication and loss analysis revealed frequent duplications and losses during Lepidoptera evolution; these duplications primarily originated through tandem and dispersed duplications and were located in syntenic regions. Transcriptomic analysis of the silkworm revealed that the groups and duplications of GH1 genes were correlated to their expression patterns, with high expression in the larval midgut and fat body. These findings suggest that GH1 gene duplications and losses and expression have played a significant role in Lepidopteran adaptation to diverse host plants. Overall, this study provides comprehensive insights into the evolutionary trajectories of GH1 genes, highlighting their potential contribution to insect–plant interactions in Lepidoptera. Full article

The present investigation systematically elucidates the distinct functional specialization within the M1–M3 midgut sections of the significant agricultural pest, Nezara viridula. Employing an integrated transcriptomic analysis, three pivotal discoveries were achieved: (1) each midgut segment possesses unique gene expression signatures; (2) metabolic and signal transduction pathways exhibit coordinated regulatory patterns; and (3) parallel expression changes occur between neuroreceptor (e.g., TACR/HTR) and metabolic enzyme (e.g., GLA/NAGA) genes within identical midgut segments. These data reveal that the M1 region is primarily enriched in metabolic processes and neural signaling; the M2 region emphasizes cellular junctions and immune responses, while the M3 region is mainly responsible for cellular senescence and renewal. These discoveries advance the understanding of feeding adaptation mechanisms in Hemipteran insects and propose a “metabolism–defense–regeneration” functional model for the midgut. The established multi-level analytical framework provides a robust methodology for subsequent dissection of complex biological systems, identification of key molecular targets for functional validation, and for the development of novel pest control strategies. Full article

An 8-week rearing trial was designed to estimate the dietary protein requirement and evaluate the effects of dietary protein on growth performance, plasma parameters, and digestive enzyme activities of blunt snout bream at two growth stages. Six practical diets were prepared to feed two sizes of fish (larger fish: initial weight of 153.69 ± 0.85 g; smaller fish: initial weight of 40.89 ± 0.28 g) with graded protein levels (26%, 28%, 30%, 32%, 34%, and 36%). Our results show that the final weight, weight gain (WG), and specific growth rate (SGR) of the fish initially rose to peak values and then declined as the dietary protein levels increased. The higher WG and SGR were recorded in the larger fish fed diets containing 30%, 32%, and 34% protein, and in the smaller fish fed a 30% protein diet, all significantly higher than those in the control group (p < 0.05). No significant differences were observed in the feed conversion ratio (FCR), viscerosomatic ratio (VR), hepatosomatic index (HSI), condition factor (CF), or survival rate among the treatments at both growth stages (p > 0.05). The plasma total protein (TP) content was highest at both growth stages in fish fed a 30% protein diet (p < 0.05). As the dietary protein level increased, the plasma urea content of the larger fish increased, peaked in the 34% protein group (p < 0.05), and then remained stable. In contrast, no significant difference in the plasma urea content was seen among the treatment groups of the smaller fish (p > 0.05). Protease activity in the fish foregut at both growth stages peaked in the 32% protein group (p < 0.05). In the midgut of the larger fish, protease activity was higher in the control group, while in the smaller fish, it was higher in the 36% protein group (p < 0.05). In the larger fish, hindgut protease activity was higher in the 34% protein group (p < 0.05), while in the smaller fish, there was no significant difference in the hindgut protease activity among all groups (p > 0.05). The dietary protein levels had no significant effect on lipase activity in the foregut, midgut, or hindgut, or on amylase activity in the foregut or midgut of the fish at the two growth stages (p > 0.05). However, hindgut amylase activity was highest in the control group of the smaller fish (p < 0.05). Based on regression analysis, the optimal dietary protein levels for the larger and smaller fish were 30.45% and 29.95%, respectively. Overall, appropriate dietary protein levels (30%) could improve the growth performance, immune function, and health status of fish at two growth stages and promote the adaptive response of their digestive system, especially the spatial regulation of protease activity in different gastrointestinal regions. Full article

This study presents the second phase of a year-long investigation comparing multiple PCR analyses and histological examinations to confirm the presence of characteristic lesions of each pathogen in three different regions of Latin America. More than 20 agents, including DNA and RNA viruses, bacteria and microsporidia, have been targeted. In addition to wild Penaeus vannamei, which was studied previously, samples of wild P. stylirostris and P. monodon were included. Notably, a positive PCR test result alone does not confirm the presence of a viable pathogen or a disease state. Similarly, positive PCR results do not necessarily correlate with the presence of histological lesions characteristic of the targeted pathogen. Wenzhou shrimp virus 8 (WzSV8) was found to be widespread among shrimp in all regions, including both farm-raised and wild populations. Histopathological analysis indicated that shrimp typically presented coinfections, such as WzSV8, Decapod hepanhamaparvovirus (DHPV), chronic midgut inflammation, and tubule distension/epithelial atrophy, consistent with the toxicity of Pir A/B or another bacterial toxin. Bacterial muscle necrosis was also found in some regions. In general, bacterial infection was the dominant pathology in all three regions during the year. We also postulate that both WzSV8 and DHPV can infect not only hepatopancreatic cells but also cells in the ceca and intestine. Full article

Rift Valley fever phlebovirus (RVFV) is a zoonotic mosquito-borne pathogen endemic to sub-Saharan Africa and the Arabian Peninsula which causes Rift Valley fever in ruminant livestock and humans. Co-infection with divergent viral strains can produce reassortment among the L, S, and M segments of the RVFV genome. Reassortment events can produce novel genotypes with altered virulence, transmission dynamics, and/or mosquito host range. This can have severe implications in areas where RVFV is endemic and convolutes our ability to anticipate transmission and circulation in novel geographic regions. Previously, we evaluated the frequency of RVFV reassortment in a susceptible ruminant host and observed low rates of reassortment (0–1.7%). Here, we tested the hypothesis that reassortment occurs predominantly in the mosquito using a highly permissive vector, Culex tarsalis. Cells derived from Cx. tarsalis or adult mosquitoes were co-infected with either two virulent (Kenya-128B-15 and SA01-1322) or a virulent and attenuated (Kenya-128B-15 and MP-12) strain of RVFV. Our results showed approximately 2% of virus genotypes isolated from co-infected Cx. tarsalis-derived cells were reassortant. Co-infected mosquitoes infected via infectious bloodmeal resulted in a higher percentage of reassortant virus (2–60%) isolated from midgut and salivary tissues at 14 days post-infection. The percentage of reassortant genotypes isolated from the midguts of mosquitoes co-infected with Kenya-128B-15 and SA01-1322 was similar to that of mosquitoes co-infected with Kenya-128B-15 and MP-12- strains (60 vs. 47%). However, only 2% of virus isolated from the salivary glands of Kenya-128B-15 and SA01-1322 co-infected mosquitoes represented reassortant genotypes. This was contrasted by 54% reassortment in the salivary glands of mosquitoes co-infected with Kenya-128B-15 and MP-12 strains. Furthermore, we observed preferential inclusion of genomic segments from the three parental strains among the reassorted viruses. Replication curves of select reassorted genotypes were significantly higher in Vero cells but not in Culex—derived cells. These data imply that mosquitoes play a crucial role in the reassortment of RVFV and potentially contribute to driving evolution of the virus. Full article

The primary mode of transmission for Chagas disease is vector-borne transmission, spread by hematophagous insects of the Triatominae subfamily. In Mexico, the triatomine Meccus pallidipennis is particularly significant in the transmission of Trypanosoma cruzi. This study focused on analyzing protein expression and modifications by glycosylation in different regions of the digestive tract of fifth-instar nymphs of M. pallidipennis. Two gut sections were dissected and extracted: the anterior midgut (AMG) and the proctodeum or rectum (RE). Proteins were extracted from each tissue sample and profiled by one- and two-dimensional electrophoresis; protein glycosylation was analyzed by lectin affinity. Our results showed significant differences in protein expression and glycosylation between both gut regions, with modifications being more frequent in the RE. The proteins HSP70, actin, and tubulin were analyzed, finding a differential expression of the latter two between AMG and RE. Understanding glycosylation patterns provides critical insights into vector–pathogen interactions that could eventually inform novel control approaches. Furthermore, the potential use of lectins as insecticidal agents highlights the broader implications of glycoprotein research in the future development of strategies on vector control to disrupt T. cruzi transmission. Full article

Chitin deacetylases (CDAs) are carbohydrate esterases associated with chitin metabolism and the conversion of chitin into chitosan. Studies have demonstrated that chitin deacetylation is essential for chitin organization and compactness and therefore influences the mechanical and permeability properties of chitinous structures, such as the peritrophic membrane (PM) and cuticle. In the present study, two genes (ApCDA5a and ApCDA5b) encoding CDA protein isoforms were identified and characterized in Chinese oak silkworm (Antheraea pernyi) larvae. Although five signature motifs were identified, CDA5 proteins only have the chitin-deacetylated catalytic domain. Spatiotemporal expression pattern analyses revealed that both transcripts presented the highest abundance in the anterior region of the midgut during the feeding period after molting, suggesting their role in chitin turnover and PM assembly. The down-regulation of ApCDA5a and ApCDA5b via RNA interference (RNAi) was correlated with the breakage of chitin microfibrils in the PM, suggesting that group V CDAs were essential for the growth and assembly of the chitinous layer. Additionally, ApCDA5a and ApCDA5b may have non-overlapping functions that regulate the morphological characteristics of PM chitin construction in different ways. Larvae injected with double-stranded RNA (dsRNA) against ApCDA5a and ApCDA5b transcripts were less resistant to infection by N. pernyi than those in the control groups. These results revealed that down-regulating ApCDA5a and ApCDA5b had independent effects on the PM structure and undermined the intactness of the PM, which disrupted the function of the PM against microsporidia infection per os. Our data provide new evidence for differentiating CDA functions among group V CDAs in lepidopteran insects. Full article

Background: Gastrointestinal neuroendocrine neoplasms (GI-NENs) represent a diverse group of tumors, with surgical resection being the gold standard for treatment. Materials and Methods: A retrospective analysis was conducted on 63 patients (32 women, 31 men) who underwent surgery for GI-NENs at the Department of Digestive Tract Surgery from January 2013 to June 2023. Tumors were classified by stage (localized, regionally advanced, metastatic). Results: Clinical symptoms were reported by 42 (66.7%) patients, with abdominal pain being the most common symptom, affecting 28 (44.4%) patients. The majority of tumors (44, 69.8%) originated in the midgut. The most frequently performed surgery was right hemicolectomy, carried out on 33 (52.4%) patients. Radical tumor resection was performed in 35 (55.6%) patients. Postoperative complications occurred in 12 (19%) patients, with male gender identified as an independent predictive factor for complications (p = 0.04). Non-functioning tumors were more common (33, 52.4%), and most tumors were classified as grade 1 histopathologically (49, 77.8%). Distant metastases were present in 29 (46%) patients. The overall two-year survival rate was 94.9%, with a five-year survival rate also estimated at 94.9%. Conclusions: GI-NENs are often diagnosed at advanced stages, frequently with distant or lymph node metastases, and predominantly arise in the midgut. Despite low postoperative morbidity and mortality, male gender may be a predictor of postoperative complications. Overall, the prognosis for GI-NENs is favorable, reflected in high overall survival rates. Full article

Drosophila Hey is a basic helix–loop–helix–orange (bHLH-O) protein with an important role in the establishment of distinct identities of postmitotic cells. We have previously identified Hey as a transcriptional target and effector of Notch signalling during the asymmetric division of neuronal progenitors, generating neurons of two types, and we have shown that Notch-dependent expression of Hey also marks a subpopulation of the newborn enteroendocrine (EE) cells in the midgut primordium of the embryo. Here, we investigate the transcriptional regulation of Hey in neuronal and intestinal tissues. We isolated two genomic regions upstream of the promoter (HeyUP) and in the second intron (HeyIN2) of the Hey gene, based on the presence of binding motifs for Su(H), the transcription factor that mediates Notch activity. We found that both regions can direct the overlapping expression patterns of reporter transgenes recapitulating endogenous Hey expression. Moreover, we showed that while HeyIN2 represents a Notch-dependent enhancer, HeyUP confers both Notch-dependent and independent transcriptional regulation. We induced mutations that removed the Su(H) binding motifs in either region and then studied the enhancer functionality in the respective Hey mutant lines. Our results provide direct evidence that although both enhancers support Notch-dependent regulation of the Hey gene, their role is redundant, as a Hey loss-of-function lethal phenotype is observed only after deletion of all their Su(H) binding motifs by CRISPR/Cas9. Full article

Colorectal cancer remains a major global health concern. Colonoscopy, the gold-standard colorectal cancer diagnostic, relies on the visual detection of lesions and necessitates invasive biopsies for confirmation. Alternative diagnostic methods, based on nanomedicine, can facilitate early detection of malignancies. Here, we examine the uptake of surface-enhanced Raman scattering nanoparticles (SERS NPs) as a marker for intestinal tumor detection and imaging using an established Drosophila melanogaster model for gut disease. Young and old Oregon-R and w¹¹¹⁸ flies were orally administered SERS NPs and scanned without and upon gut lumen clearance to assess nanoparticle retention as a function of aging. Neither young nor old flies showed significant NP retention in their body after gut lumen clearance. Moreover, tumorigenic flies of the esg-Gal4/UAS-Ras^V12 genotype were tested for SERS NP retention 2, 4 and 6 days after Ras^V12 oncogene induction in their midgut progenitor cells. Tumorigenic flies showed a statistically significant NP retention signal at 2 days, well before midgut epithelium impairment. The signal was then visualized in scans of dissected guts revealing areas of NP uptake in the posterior midgut region of high stem cell activity. Full article

Haemaphysalis longicornis is a common tick species that carries several pathogens. There are few reports on the influence of different hosts on the structure of midgut microflora in H. longicornis. In this study, midgut contents of fully engorged female H. longicornis were collected from the surface of tiger (Panthera tigris) and deer (Dama dama). The bacterial genomic DNA of each sample was extracted, and the V3–V4 regions of the bacterial 16S rRNA were sequenced using the Illumina NovaSeq sequencing. The diversity of the bacterial community of the fully engorged female H. longicornis on the surface of tiger was higher than that of deer. In total, 8 phyla and 73 genera of bacteria annotations were detected in the two groups. At the phylum level, the bacterial phyla common to the two groups were Proteobacteria, Firmicutes, and Actinobacteriota. At the genus level, there were 20 common bacterial genera, among which the relative abundances of Coxiella, Morganella, Diplorickettsia, and Acinetobacter were high. The Morganella species was further identified to be Morganella morganii. The alpha diversity index indicated that the bacterial diversity of the tiger group was higher than that of the deer group. Bacteroidota, Patescibacteria, Desulfobacterota, Verrucomicrobiota, and Cyanobacteria were solely detected in the tiger group. A total of 52 bacterial genera were unique in the tiger group, while one bacterial genus was unique in the deer group. This study indicates that there are differences in the structure of the gut bacteria of the same tick species among different hosts. Further culture-based methods are needed to provide a more comprehensive understanding of the tick microbiota parasitizing different hosts. Full article

The mosquito microbiota is a critical determinant of mosquito life history. It is therefore a target for novel vector control strategies like paratransgenesis. However, the microbiota in Anopheles funestus, a major African malaria vector, is poorly characterized. Thus, the study aimed to investigate the overall bacterial landscape in the salivary glands, ovaries and midguts of three laboratory strains of An. funestus differing in insecticide-resistant phenotype by sequencing the V3–V4 hypervariable region of bacterial 16S rRNA genes. When examining alpha diversity, the salivary glands harbored significantly more bacteria in terms of species richness and evenness compared to ovaries and midguts. On the strain level, the insecticide-susceptible FANG strain had significantly lower bacterial diversity than the insecticide-resistant FUMOZ and FUMOZ-R strains. When looking at beta diversity, the compositions of microbiota between the three tissues as well as between the strains were statistically different. While there were common bacteria across all three tissues and strains of interest, each tissue and strain did exhibit differentially abundant bacterial genera. However, overall, the top five most abundant genera across all tissues and strains were Elizabethkingia, Acinetobacter, Aeromonas, Cedecea and Yersinia. The presence of shared microbiota suggests a core microbiota that could be exploited for paratransgenesis efforts. Full article

By 2013, it had been shown that the genes cadherin-like receptor (Cad) and ATP-binding cassette transporter subfamily C2 (ABCC2) were responsible for insect resistance to several Cry1A toxins, acting as susceptibility-determining receptors, and many review articles have been published. Therefore, this review focuses on information about receptors and receptor-binding sites that have been revealed since 2014. Since 2014, studies have revealed that the receptors involved in determining susceptibility vary depending on the Cry toxin subfamily, and that binding affinity between Cry toxins and receptors plays a crucial role. Consequently, models have demonstrated that ABCC2, ABCC3, and Cad interact with Cry1Aa; ABCC2 and Cad with Cry1Ab and Cry1Ac; ABCC2 and ABCC3 with Cry1Fa; ABCB1 with Cry1Ba, Cry1Ia, Cry9Da, and Cry3Aa; and ABCA2 with Cry2Aa and Cry2Ba, primarily in the silkworm, Bombyx mori. Furthermore, since 2017, it has been suggested that the binding sites of BmCad and BmABCC2 on Cry1Aa toxin overlap in the loop region of domain II, indicating that Cry toxins use various molecules as receptors due to their ability to bind promiscuously in this region. Additionally, since 2017, several ABC transporters have been identified as low-efficiency receptors that poorly induce cell swelling in heterologously expressing cultured cells. In 2024, research suggested that multiple molecules from the ABC transporter subfamily, including ABCC1, ABCC2, ABCC3, ABCC4, ABCC10, and ABCC11, act as low-efficiency receptors for a single Cry toxin in the midgut of silkworm larvae. This observation led to the hypothesis that the presence of such low-efficiency receptors contributes to the evolution of Cry toxins towards the generation of highly functional receptors that determine the susceptibility of individual insects. Moreover, this evolutionary process is considered to offer valuable insights for the engineering of Cry toxins to overcome resistance and develop countermeasures against resistance. Full article

Recent studies have suggested that ABC transporters are the main receptors of Cry toxins. However, the receptors of many Cry toxins have not been identified. In this study, we used a heterologous cell expression system to identify Bombyx mori ABC transporter subfamily C members (BmABCCs) that function as receptors for five Cry toxins active in Lepidopteran insects: Cry1Aa, Cry1Ca, Cry1Da, Cry8Ca, and Cry9Aa. All five Cry toxins can use multiple ABCCs as low-efficiency receptors, which induce cytotoxicity only at high concentrations. Surface plasmon resonance analysis revealed that the K_D values between the toxins and BmABCC1 and BmABCC4 were 10⁻⁵ to 10⁻⁹ M, suggesting binding affinities 8- to 10,000-fold lower than those between Cry1Aa and BmABCC2, which are susceptibility-determining receptors for Cry1Aa. Bioassays in BmABCC-knockout silkworm strains showed that these low-efficiency receptors are not involved in sensitivity to Cry toxins. The findings suggest that each family of Cry toxins uses multiple BmABCCs as low-efficiency receptors in the insect midgut based on the promiscuous binding of their receptor-binding regions. Each Cry toxin seems to have evolved to utilize one or several ABC transporters as susceptibility-determining receptors. Full article

The highly prevalent pest Alphitobius diaperinus (Coleoptera: Tenebrionidae) causes significant structural damage in poultry farms. Despite previous investigations on its carriage of pathogenic microorganisms, our understanding of its microbiome remains limited. This study aimed to analyze the diversity of culturable gut microbiota in A. diaperinus obtained from laboratory breeding. Fifteen seventh instar larvae underwent a 24-h starvation period, followed by surface disinfection. Dissected midguts were homogenized and plated on nutrient agar (NA), brain heart infusion agar (BHI), and Bacillus cereus agar (BC). The cultured isolates were subjected to gram staining, phylogenetic analysis, biochemical property evaluation, and metabolic activity assessment. Bacterial counts were higher in BHI (2.51 × 10⁵ CFU/gut) than in NA (2.25 × 10⁵ CFU/gut), possibly due to nutrient richness. NA exhibited a dominant colony morphology of gram-negative bacilli, while BHI displayed additional distinct colonies of gram-positive cocci. Surprisingly, yeast-like colonies were observed on BC plates. Based on 16S rRNA gene sequences, eight bacterial isolates were identified as Enterobacter sp., and two as Staphylococcus sp. Using RNA gene ITS region sequences, two yeast isolates were identified as Debaryomyces sp. and Hyphopichia sp. A preliminary species-level identification of bacteria (Enterobacter cloacae, Staphylococcus gallinarum, and Staphylococcus succinus) was achieved using API systems and complementary biochemical tests. Discrepancies between phylogenetic analysis and phenotypic data suggest the potential existence of new species or subspecies. Further comprehensive studies are required to confirm this hypothesis. Full article