The tri-spine horseshoe crab, Tachypleus tridentatus, is an evolutionarily ancient marine chelicerate of considerable ecological and conservation importance in the Indo-West Pacific. Although several mitochondrial genomes have been reported, comprehensive comparative mitogenomic analyses encompassing all extant limulid genera remain limited. In this study, we sequenced and characterized the complete mitochondrial genome of an F1 Tachypleus tridentatus individual derived from wild-caught broodstock from the Beibu Gulf, northern South China Sea, and subsequently maintained at an aquaculture facility, using high-throughput sequencing, and performed an integrated comparative mitogenomic analysis across the three extant genera of Limulidae. The circular mitochondrial genome was 15,006 bp in length and contained the typical set of 37 mitochondrial genes, including 13 protein-coding genes (PCGs), 22 transfer RNA genes, and two ribosomal RNA genes. The newly sequenced Tachypleus tridentatus mitogenome showed highly conserved genome organization and nucleotide composition relative to previously published conspecific mitogenomes, confirming established mitochondrial characteristics of the species. The principal contribution of this study is the addition of a complete Beibu Gulf mitogenome and its integration with 12 additional complete mitochondrial genomes representing all extant limulid species, together with four mitochondrial markers, K2P genetic-distance analyses, and independent BI and ML phylogenetic reconstruction. The concordance among these mitochondrial datasets provides an integrated assessment of genetic divergence and mitochondrial phylogenetic relationships within Limulidae and expands the geographic representation of complete mitochondrial genomic resources for Tachypleus tridentatus.

The sarcopenia burden is escalating in super-aged societies. The “gut-muscle axis” hypothesis suggests gut dysbiosis contributes to muscle wasting via inflammation and metabolic dysregulation, yet identifying consistent markers is complicated by heterogeneity. This study determined prevalence, risk factors, and microbiota signatures in a homogeneous cohort of community-dwelling men aged ≥ 75 receiving chronic disease management in Shanghai. We employed a cross-sectional design, enrolling 172 older men (mean age 82 ± 3 years). Sarcopenia was diagnosed using AWGS 2019 criteria. Comprehensive clinical data, including serological markers and medication history, were collected. Fecal samples from a subgroup (n = 45) underwent 16 S rRNA sequencing. Clinical risk factors were identified using backward stepwise multivariate logistic regression with Bootstrap internal validation. Multivariate logistic regression and Linear Discriminant Analysis Effect Size (LEfSe) were used to identify independent factors. To mitigate potential overfitting and small-sample bias in the microbiota analysis, complementary differential abundance testing (e.g., ANCOM-BC) and exploratory penalized multivariable modeling (LASSO feature selection and Firth’s penalized regression) were strictly applied. Sarcopenia prevalence was 25.6

Bellamya limnophila is a mollusk of significant medical and economic value in China. Understanding the complete mitochondrial genome of this species will better establish a foundation for systematic classification research on Viviparidae. Therefore, we sequenced the complete mitochondrial genome of B. limnophila, conducted a comprehensive analysis of its structural characteristics, and constructed a phylogenetic tree using maximum likelihood and Bayesian methods. The results showed that the genome sequence is 16,991 bp in length, including 13 protein-coding genes (PCGs), 20 transfer RNA genes (tRNAs), 2 ribosomal RNA genes (rRNAs), and 1 non-coding region (D-loop). In summary, the Ka/Ks ratios of all PCGs were <1, indicating that purifying selection dominated the evolutionary process of these snails. The entire genome structure exhibited conservative features, such as the majority of start codons being the standard ATG codon and the majority of tRNA genes having the standard cloverleaf secondary structure. B. limnophila and B. quadrata showed collinearity in terms of sequence homology. Phylogenetic analysis indicates that the clade formed by the genera Margarya, Cipangopaludina, and Bellamya is the sister group of the genus Viviparus; Bellamya limnophila is more closely related to B. quadrata than to other species. This study contributes to the mitochondrial genome database of the family Viviparidae and provides valuable insights into the phylogenetic relationships of related snails.

Human norovirus (HuNoV) is the leading foodborne virus worldwide, causing acute gastroenteritis and imposing significant public health and economic burdens. Nevertheless, because viral titers are often low in environmental and food samples, enriching viral particles thereof presents challenges. This study aimed to establish a rapid and efficient enrichment strategy for HuNoV using recombinant oyster heat shock protein 70-conjugated magnetic beads (roHSP70-MBs). The enrichment protocol was optimized by evaluating protein coating concentration, bead-to-sample volume ratio, and incubation time based on GII.4 clinical samples. The optimal conditions (100.0 mu g/mL roHSP70, 1:10 bead-to-sample volume ratio, and 45 min incubation at 37 degrees C) were applied to enrich six HuNoV genotypes (i.e., GI.3, GII.2, GII.3, GII.4, GII.12, and GII.17). Under these conditions, roHSP70-MBs demonstrated a 10-fold increase in sensitivity over porcine gastric mucin-conjugated magnetic beads (PGM-MBs), achieving a lower limit of quantification of 1-18 copies/mL in the eluate. In artificially contaminated food matrices, roHSP70-MBs showed significantly higher recovery rates than PGM-MBs and polyethylene glycol precipitation, with rates of 31-37 % in strawberries, 67-78 % in lettuce, and 37-42 % in oysters. Stability studies indicated that roHSP70-MBs without any protein protectant stored at 4 degrees C exposed-to-light retained over 70 % of their initial enrichment efficiency for 56 days. By combining rapid viral enrichment with thermal lysis, the current method reduced total processing time to approximately 2 h and eliminated the need for commercial RNA extraction kits, offering a promising strategy for the enrichment of HuNoV particles in complex food matrices.

Norovirus is a major cause of acute viral gastroenteritis in humans. Molecular biology-based detection methods play a pivotal role in ensuring accurate and specific diagnosis. The inclusion of Qβ phage particles as armored positive controls in these assays can further enhance their reliability and specificity. Herein, we discuss rational design strategies to improve the stability of Qβ bacteriophage capsid proteins armored with RNA using Discovery Studio 2019 protein design software. Amino acid mutation sites were deter-mined based on changes in folding free energy differences (ΔΔGmut). These single-site mutations were subsequently evaluated using molecular dynamics simulations. Wild-type and mutant recombinant expression plasmids were constructed and transformed into Escherichia coli BL21 (DE3) for cloning and expression. The stability of Qβ virus-like particles (VLPs) was assessed using real-time fluorescence RT-qPCR. The results showed that structurally intact and uniformly distributed wild-type and single-site mutant VLPs were successfully obtained. Stability analyses indicated that at 4 °C, 25 °C, 37 °C, 45 °C, and 60 °C, the single-site mutant exhibited a significantly lower rate of degradation than the wild-type. In conclusion, rational design enables the generation of single-site mutant VLPs with enhanced stability, providing a safer and more stable standard reference material for the molecular detection of foodborne viruses.

Functional constipation is a prevalent gastrointestinal disorder associated with gut microbiota dysbiosis. Probiotics have emerged as a promising strategy to restore microbial balance, particularly strains capable of degrading dietary fibres like pectin. In this study, two indigenous pectin-degrading strains, Lactiplantibacillus plantarum PD-14 and Lactiplantibacillus pentosus PD-16, which were isolated from fermented tofu, were used to facilitate the decomposition of dietary fiber in the intestinal tract, and to modulate intestinal microbiota composition and metabolic activity. Whole-genome sequencing revealed the presence of pectate lyase 3 family pectate lyase genes in both strains, establishing their intrinsic capacity for dietary fibre utilization. In a loperamide-induced constipation murine model, both strains exhibited a favorable safety profile and alleviated constipation symptoms. Notably, the two strains exhibited distinct regulatory profiles. L. plantarum PD-14 increased fecal short-chain fatty acids, particularly acetic acid and propionic acid, accompanied by an enrichment of Bifidobacterium. In contrast, L. pentosus PD-16 shortened whole-gut transit time and reduced the abundance of constipation-associated taxa, such as Faecalibaculum. Unlike the drug PEG-4000, both probiotic strains successfully changed the gut microbiota. These findings demonstrated that indigenous, pectin-degrading microbes alleviate constipation through strain-specific mechanisms, supporting the development of regionally adapted probiotics for precise health management.

Given the resilience of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on frozen food, there is a risk that contaminated products could serve as vectors for viral transmission. Yet, methods capable of inactivating the virus at sub-zero temperatures without compromising the food's taste and quality are scarce. The high-pressure environment that arises spontaneously during isochoric freezing has demonstrated efficacy in suppressing or inactivating harmful microorganisms, such as bacteria; however, its effectiveness against coronavirus and the mechanisms involved remain unclear. In this study, we employed Porcine epidemic diarrhea virus (PEDV) as a proxy for SARS-CoV-2 to examine the effects of isochoric freezing on PEDV infectivity and to evaluate post-treatment alterations in the integrity of viral nucleic acids and envelopes, as well as changes in antigenic properties. Our experimental findings indicate that after a 6-h isochoric freezing treatment at -20 °C and 201 MPa, the titer decreased by 1.18 log10(TCID50/mL). While the viral nucleic acid remained intact post-treatment, the envelope's integrity was significantly impaired, accounting for the loss of infectivity. Moreover, the antigenic properties of the virus showed a slight increase following isochoric freezing treatment. This study pioneers the exploration of isochoric freezing technology's inactivation effects on coronaviruses, including preliminary mechanisms, offering novel perspectives for managing coronavirus contamination in frozen food and highlighting the potential of isochoric freezing in vaccine inactivation processes.

Human norovirus (HuNoV) is the leading cause of acute nonbacterial gastroenteritis globally. Histo-blood group antigens (HBGAs) have been recognized as attachment factors for HuNoV. However, the receptors or other potential attachment factors for HuNoV have not been fully elucidated. A bacterial cell surface-displayed system was used to express GII.4 HuNoV protruding domain (GII.4 P) to capture attachment factors from a diarrheal sample of a patient infected with GII.4 HuNoV. A library of candidate HuNoV proteinaceous attachment factors was constructed. Through a database comparison, a literature review, the predicted protein subcellular localization, the protein-protein interactions (PPIs) network analysis, the molecular simulation and the protein docking analysis, four candidate attachment factors were selected for further investigation from the initial 118 candidates. Finally, two proteins, namely myelin-oligodendrocyte glycoprotein (MOG) and leucine-rich repeat-containing protein 15 (LRRC15), were demonstrated strongly binding to GII.4 P by both in vivo and in vitro assays. The binding between MOG/LRRC15 and GII.4 P can be blocked by GII.4 HuNoV antibody. Immunofluorescence showed that MOG/LRRC15 and GII.4 HuNoV co-localized in the cytoplasm in transfected cells. However, GII.4 HuNoV did not proliferate in the newly constructed MOG/LRRC15 overexpressing cells. This study describes attempts to identify HuNoV proteinaceous attachment factors from the diarrheal sample. Findings from this study will aid in understanding of HuNoV infection in humans, enrichment of GII.4 HuNoV from various environments and construction of cell lines for HuNoV cultivation in vitro.

Understanding the processes that drive microbial community assembly is a fundamental question in ecology, with important implications for predicting community responses to environmental disturbances. River ecosystems are under growing pressure from human disturbances, jeopardizing their ecological functions. Here, we investigated bacterial and protistan communities along the Yujiang River using environmental DNA metabarcoding. Bacterial communities exhibited significantly greater alpha diversity and broader habitat niches compared to protists. Additionally, a negative correlation was found between alpha diversity and niche breadth for both groups. Protistan communities exhibited significantly higher beta diversity (Bray–Curtis distance) than bacterial communities, with species turnover being the principal factor driving the variations in both communities. Null model results indicated that heterogeneous selection primarily structured bacterial communities, while stochastic processes (drift) mainly governed protist communities. Redundancy analysis and Mantel tests showed significant associations between environmental factors (e.g., temperature and pH) and bacterial community composition. Moreover, the longitude of sampling sites was linked to spatial variations in both bacterial and protistan communities. Further analyses, including distance-decay patterns, variation partitioning, and multiple regression on distance matrices, demonstrated that bacterial communities were driven by both environmental and spatial factors, while protist communities exhibited a stronger response to spatial factors. These results enhance our understanding of microbial community assembly in river ecosystems and provide valuable insights for the conservation and sustainable management of freshwater systems.

Human norovirus (HuNoV), a primary cause of foodborne illness, is frequently transmitted through contaminated berries. Serbia is a global producer of raspberries and other berries, yet data on HuNoV prevalence and genogroup diversity are limited. This study aimed to assess the prevalence, viral load, and genotypes of HuNoV circulating in raspberries and blackberries marketed in Serbia. A total of 450 berry samples were collected in 2023 from orchards, cold storage facilities, local markets, and roadside vending stands. Norovirus RNA was extracted using a modified ISO 15216-2 protocol. RT-qPCR positive samples were subsequently quantified by digital RT-PCR (RT-dPCR). Genotyping employed next-generation sequencing (NGS) of genome encoding major and minor capsid proteins, supplemented by Sanger sequencing. Nineteen samples (4.2%) tested positive for HuNoV, including four GI and fifteen GII strains, with higher prevalence in frozen (11.1%) than fresh berries (2.0%). Viral loads ranged from 34–105 gc/g for GI and 23–658 gc/g for GII, with medians of 72 and 153 gc/g, respectively. Genotypes GI.6, GII.2, GII.4, and GII.7 were identified, each represented by more than two sublineages and multiple introduction events detected by phylogenetic analysis. RT-dPCR provided precise quantification, while NGS enabled genotype characterization, together supporting enhanced surveillance, risk assessment, and management of norovirus contamination in berries destined for domestic and international markets.

Understanding the biodiversity of aquatic communities and the underlying mechanisms that shape biodiversity patterns and community dynamics is crucial for the effective conservation and management of freshwater ecosystems. However, traditional survey methods often fail to comprehensively capture species diversity, particularly for low-abundance taxa. Moreover, studies integrating both metazoan and fish communities at fine spatial scales remain limited. To address these gaps, we employed a multi-marker eDNA metabarcoding approach, targeting both the 12S and 18S rRNA gene regions, to comprehensively investigate the composition of metazoan and fish communities in the Yujiang River. A total of 12 metazoan orders were detected, encompassing 15 families, 21 genera, and 19 species. For the fish community, 32 species were identified, belonging to 25 genera, 10 families, and 7 orders. Among these, Adula falcatoides and Coptodon zillii were identified as the most prevalent and abundant metazoan and fish species, respectively. Notably, the most prevalent fish species, C. zillii and Oreochromis niloticus, are both recognized as invasive species. The Bray–Curtis distance of metazoa (average: 0.464) was significantly lower than that of fish communities (average: 0.797), suggesting higher community heterogeneity among fish assemblages. Beta-diversity decomposition indicated that variations in the metazoan and fish communities were predominantly driven by species replacement (turnover) (65.4% and 70.9% for metazoa and fish, respectively) rather than nestedness. Mantel tests further revealed that species turnover in metazoan communities was most strongly influenced by water temperature, while fish community turnover was primarily affected by water transparency, likely reflecting the physiological sensitivity of metazoans to thermal gradients and the dependence of fish on visual cues for foraging and habitat selection. In addition, a co-occurrence network of metazoan and fish species was constructed, highlighting potential predator-prey interactions between native species and Corbicula fluminea, which emerged as a potential keystone species. Overall, this study demonstrates the utility of multi-marker eDNA metabarcoding in characterizing aquatic community structures and provides new insights into the spatial dynamics and species interactions within river ecosystems.

Human noroviruses (HuNoVs) are the primary cause of viral gastroenteritis globally. Nevertheless, available anti-HuNoV approaches remain limited. The current study revealed that a Lactobacillus rhamnosus strain, that is, L. rhamnosus YQ001, originated de Man, Rogosa and Sharpe (MRS)-based fermentation broth (FB) and cell-free supernatant (CFS) could significantly inhibit the replication of GII.4 HuNoVs in zebrafish larvae (Danio rerio), reducing viral RNA titers by approximately 2.18 log10 and 1.12 log10 copies, respectively. In addition, the inhibitory effect of FB was significantly stronger than that of CFS (P < 0.05), while L. rhamnosus YQ001 alone demonstrated no inhibitory effect. Zebrafish larvae injected with FB-treated GII.4 HuNoVs demonstrated reduced immune responses (i.e., significantly decreased upregulation of innate immune genes, ifn and mx, P < 0.05), compared to those of larvae injected with untreated GII.4 HuNoVs. Additionally, in situ capture RT-qPCR and enzyme-linked immunosorbent assay suggested that membrane proteins, especially C2JVE6, C2JX39 and C2K0J4, in L. rhamnosus YQ001, could bind to GII.4 HuNoVs. Altogether, the current study demonstrated the inhibitory effect of L. rhamnosus YQ001 originated FB and CFS on GII.4 HuNoVs in zebrafish larvae and identified the binding capacity of membrane proteins from L. rhamnosus YQ001. Collectively, these results highlight the synergistic effect of CFS and cell membrane proteins in GII.4 HuNoV control. IMPORTANCE:Human noroviruses (HuNoVs) are the leading cause of viral gastroenteritis globally, yet effective antiviral treatments remain limited. The current study demonstrated that Lactobacillus rhamnosus YQ001 could inhibit GII.4 HuNoV replication in zebrafish larvae. The cell-free supernatant and membrane proteins originated from L. rhamnosus YQ001 did work synergistically in GII.4 HuNoVs control. The membrane proteins could bind to the viral capsid. These findings offer a unique insight into the antiviral mechanisms of L. rhamnosus YQ001, laying the groundwork for developing probiotic-based foods to anti-HuNoVs.

The objective of the current study was to investigate and compare the impact of enzymatic hydrolysis and pre-fermentation with probiotics on the biological activity and metabolic profile of melon juice. We found that fermentation with Lactiplantibacillus plantarum N13, not only decreased the juice pH, total soluble solids and reducing sugar, but also promoted the release of phenolic compounds, increased antioxidant activity and improved juice's aroma profile. Although pectinase and cellulase aided in the release of reducing sugar and total phenols in the juice, the heating process involved in the enzymatic treatment decreased total soluble solids and phenolics in the juice. Altogether, both enzymatic treatment and fermentation with probiotics could aid in the release of bioactive compounds, while fermentation with probiotics demonstrated greater impact on the biological activity and metabolic profile of melon juice. The findings of the current study suggested the use of probiotic cultivars in the pre-alcohol-fermentation process.

Chronic constipation (CC) and functional constipation (FC) are common gastrointestinal disorders that significantly affect quality of life. This study investigates the intestinal microbiota characteristics in CC and FC patients, revealing microbial imbalances characterized by reduced beneficial taxa, such as Acinetobacter, Blautia, Dorea formicigenerans, Eubacterium ramulus, and Halomonas, alongside increased levels of Alistipes, Holdemanella, Parabacteroides, Pseudomonas, Streptococcus, and so on. These findings highlight dysbiosis as a critical factor in constipation pathogenesis and provide a foundation for the bottom-up design of targeted probiotics and prebiotics. Potential therapeutic strategies include tailored probiotic formulations to replenish deficient taxa and the application of prebiotics to restore microbial balance. Additionally, the role of microbial metabolites, particularly short-chain fatty acids, and the microbiota-gut-brain axis offers further insight into mechanisms underlying symptom modulation. The integration of artificial intelligence enhances precision in probiotic design, enabling the prediction of strain-specific combinations optimized for therapeutic efficacy. This microbiota-centered approach underscores the potential for personalized interventions in addressing dysbiosis and advancing innovative management for CC and FC.

Angulyagra polyzonata is a significant freshwater snail species in southern China. However, its wild resources have sharply declined due to overfishing. To assess the current status of germplasm resources in the Guangxi region, during this study, we first successfully developed nine pairs of primers that enable the amplification of highly polymorphic microsatellite markers (SSRs) with trinucleotide and tetranucleotide repeat sequences (PIC values ranging from 0.662 to 0.861) using transcriptomic data. Then, these designed primers were tested and applied for the genetic investigation of selected wild populations of the species. Finally, a genetic diversity analysis was conducted based on 12 wild populations (360 individuals) in Guangxi. After 798,244 SSR loci were screened out via high-throughput sequencing, the results showed that dinucleotide repeats accounted for the highest proportion (47.64%), mainly consisting of (AC/GT)n repeat units. Among the SSR loci in A. polyzonata, microsatellite loci with 5 to 20+ repeats are the most abundant. All nine selected and tested SSR loci significantly deviated from Hardy–Weinberg equilibrium (p < 0.001) and had heterozygote deficiency (average inbreeding coefficient of F = 0.390), indicating widespread inbreeding. The fixation index among populations was high (average Fst = 0.175), with 73% of the genetic variation occurring within populations and 27% between populations. Gene flow (Nm) was generally restricted (most population pairs had Nm < 1), with the (Tiandeng) TD and (Long’an) LA populations showing the smallest differentiation (Fst = 0.017), and the (Qinnan) QN and (Yinhai) YH populations showing the greatest differentiation (Fst = 0.409). UPGMA clustering and structure analysis (K = 2) divided the 12 populations into two subgroups. Overall, our research suggests that the genetic diversity of the wild population of A. polyzonata in the Guangxi region has declined. Thus, prioritizing the protection of highly genetically diverse populations, such as the LA population, is urgently needed. This study provides a scientific basis for the protection and sustainable utilization of A. polyzonata resources in Guangxi.

Norovirus (NoV), an important cause of human viral gastroenteritis worldwide, recognizes human histo-blood group antigens (HBGAs) as receptors. Oysters are a vector of foodborne transmission of NoV, and HBGAs have been found in oyster tissues. In this study, CgFUT1 and CgFUT2, the key genes involved in the synthesis of HBGAs in Crassostrea gigas, were successfully expressed in Pichia pastoris, and 32.6 kDa target proteins were obtained after purification, concentration, and dialysis treatments. Western blot analysis using FUT1 and FUT2 antibodies showed that CgFUT1 and CgFUT2 have antigenic similarity as human FUT1 and FUT2. Enzyme catalysis assays using Galβ1-3GlcNAc and Galβ1-4GlcNAc as substrates showed that these substrates react with GDP-Fuc to generate Fucα1-2GalβGlcNAc under the action of CgFUT1 and CgFUT2. High-resolution mass spectrometry analysis revealed that CgFUT1 and CgFUT2 have the same substrate specificity, both reacting with Galβ1-3GlcNAc and Galβ1-4GlcNAc. The results of this study demonstrate the probable role of CgFUT1 and CgFUT2 in regulating substrates for H antigen synthesis in oysters and provide a reference for future studies into the functions of these genes. The study also lays a foundation for further exploration of the molecular mechanisms underlying NoV accumulation in oysters.

Angulyagra polyzonata is an economically important mollusk in China, but detailed insights into its mitochondrial genome remain scarce. In this study, we sequenced and comprehensively analyzed the structural features and selection pressures of the A. polyzonata mitochondrial genome. The maximum likelihood method and Bayesian phylogenetic inference method were used to construct a phylogenetic tree of A. polyzonata with 21 other species, including gastropods and bivalves. The full-length mitochondrial genome of 17,379 bp was found to include 22 transfer RNA genes, 2 ribosomal RNA genes, and 13 protein-coding genes, exhibiting similarity to the composition and arrangement of mitochondrial genes in other gastropod species. Notably, the Ka/Ks ratios of mitochondrial protein-coding genes (nad5, cox3, nad3, nad2, cox1, cox2, atp8, atp6, nadl, nad6, cob, nad4l, and nad4) were <1, which indicates that the snail genes of the three genera of the family may have been subjected to strong natural selection pressure during the evolutionary process, so that the number of synonymous mutations (ks) in genes was much more than that of nonsynonymous mutations (ka). Comparative genomic analysis indicated that, apart from the absence of trnW and trnQ, the gene composition of A. polyzonata shares a high degree of homology with other members of the conical snail family. Phylogenetic analysis demonstrated that the selected species could be classified into two primary clades in which A. polyzonata clustered with the Viviparidae family. This study bridges the knowledge gap regarding the mitochondrial genome of A. polyzonata and offers valuable insights into the systematic relationships within the Viviparidae family.

Mystus guttatus, a second-class protected species in China, has undergone severe population decline due to anthropogenic and environmental pressures, yet conservation efforts are hindered by limited genomic resources and a lack of mechanistic insights into its stress response systems. Here, the first full-length transcriptome of M. guttatus was generated via SMRT sequencing. A total of 32,647 full-length transcripts were obtained, with an average length of 1783 bp. After structure and function annotation of full-length transcripts, 30,977 genes, 1670 transcription factors (TF), 918 alternative splicing (AS), and 11,830 simple sequence repeats (SSR) were identified. In order to further explore the stress resistance of M. guttatus, 93 genes belonging to the heat shock protein (HSP) family were identified and categorized into HSP70 and HSP90 subgroups. After phylogenetic analysis and selective stress analysis, it was discovered that the hsp family has suffered purifying selection and gene loss, potentially contributing to a decrease in the stress resilience and population of M. guttatus. Using protein interaction network and molecular docking tools, we observed the intricate interplay among HSPs and discovered HSP70-HOP-HSP90 interaction, which is an essential stress response mechanism. Our study sequenced the first full-length transcriptome of M. guttatus to enhance its genomic resources for its conservation and breeding and provide new insights into the future study of stress response mechanisms on M. guttatus.

Oreochromis aureus is an economically valuable fish species, but its domestication in saline environments remains unexplored, with limited reports on how salinity stress affects its physiological functions and acclimation-related mechanisms. Thus, this study collected O. aureus exposed to different salinity stress levels (0 %o , 3 %o , 7 %o , and 11 %o ) and analyzed their growth performance, histopathology, physiological functions, and transcriptome. The results showed that salinity stress had no significant effect on growth performance. Salinity stress damaged gill tissues, decreased physiological and antioxidant activities, increased osmotic and antimicrobial activities, and altered digestive functions. Comparative transcriptome analyses identified 38,910 differentially expressed genes (DEGs), of which 11,488 were common to the three comparisons. These DEGs were significantly associated with specific salinity stress response-related KEGG pathways, including Sphingolipid signaling pathway, Lysosome, Phagosome, and Focal adhesion. The present results identify 1 GO term (regulation of biological proces) in response to salinity stress. Furthermore, 15 candidate genes related to salinity stress responses and physiological functions were also identified. (e.g., TLR2, NCF2, Sptlc2, and ctsd). On the basis of GO, KEGG and STEM analyses, the data enabled the development of a mechanistic model that details how O. aureus adapts to salinity stress by regulating physiological changes. Finally, RT-qPCR assays verified the accuracy and reliability of the high- throughput sequencing results. This study enhances our understanding of O. aureus adaptive strategies under salinity stress, while also providing relevant theoretical insights into the domestication of fish under saline conditions and the mechanisms mediating adaptations to saline aquatic environments.

Human noroviruses (HuNoVs) are the leading etiological agent causing the worldwide outbreaks of acute epidemic non-bacterial gastroenteritis. Histo-blood group antigens (HBGAs) are commonly acknowledged as cellular receptors or co-receptors for HuNoVs. However, certain genotypes of HuNoVs cannot bind with any HBGAs, suggesting potential additional co-factors and attachment receptors have not been identified yet. In addition, food items, such as oysters and lettuce, play an important role in the transmission of HuNoVs. In the past decade, a couple of attachment factors other than HBGAs have been identified and analyzed from foods and microbiomes. Attachment factors exhibit potential as inhibitors of viral binding to receptors on host cells. Therefore, it is imperative to further characterize the attachment factors for HuNoVs present in foods to effectively control the spread of HuNoVs within the food chain. This review summarizes the potential attachment factors/receptors of HuNoVs in humans, foods, and microbiome.