Traditional food markets, often referred to as wet markets, live poultry markets, or farmers' markets, remain a major source of animal-origin foods and fresh produce in many low-income and middle-income countries (LMICs). These markets bring humans, domestic animals, wildlife, and the environment into close contact and have been repeatedly implicated in the emergence of zoonotic and foodborne diseases. Yet, the overall global burden, pathogen spectrum, and risk gradients within these settings have not been comprehensively quantified. We conducted a systematic review and meta-analysis of studies published between Jan 1, 2000, and Oct 31, 2025, following the PRISMA 2020 reporting guidelines. We searched major English and Chinese databases for cross-sectional or baseline studies reporting the detection of zoonotic or foodborne pathogens in traditional food markets or related retail settings. Studies were excluded if they were not conducted in traditional markets, had non-extractable data, had a sample size of fewer than 20, had intervention-related outcomes, had duplicate populations, or did not have prevalence or detection rate reporting. Data were extracted as detection records, defined as unique pathogen-sample type-market setting combinations. Study quality was assessed with the Joanna Briggs Institute tool for prevalence studies. We used random-effects meta-analysis of logit-transformed proportions to estimate pooled prevalence and 95% CIs overall and across major subgroups. Heterogeneity was assessed with I2 and τ2, and meta-regression was used to examine temporal trends and associations between study quality and prevalence. Robustness was assessed with leave-one-out analyses and small-study effects. This study is registered with PROSPERO, CRD420251267834. We included 247 studies comprising 597 detection records from 41 countries across 16 regions. The global pooled prevalence of market-source pathogens in traditional markets was 15·32% (95% CI 13·96-16·80). By pathogen kingdom, pooled prevalence was 19·41% (16·21-23·08) for bacteria, 8·40% (7·15-9·83) for viruses, and 27·05% (21·91-32·89) for parasites. Regionally, prevalence was highest in southeast Asia (23·09% [18·43-28·52]), followed by south Asia (15·52% [10·06-23·20]) and east sub-Saharan Africa (14·54% [7·13-27·40]). By market type, fish and seafood markets showed the highest pooled prevalence (37·87% [26·05-51·33]), and by sample type, the highest estimates were observed in fish and seafood, raw meat, and environmental samples. Lower estimates were observed in cooked foods, fruit and vegetables, and worker samples, suggesting a gradient from animal and environmental reservoirs towards ready-to-eat foods and humans. Meta-regression showed no significant global linear temporal trend in prevalence, whereas higher methodological quality was associated with lower reported prevalence. Leave-one-out analyses suggested that no single study substantially altered the overall findings. Traditional markets represent persistent global hotspots for zoonotic and foodborne pathogens, particularly in LMICs and high-risk food chains. The observed exposure gradient highlights crucial intervention points. These findings support a One Health-oriented framework for risk assessment and targeted surveillance, while emphasising the need for harmonised, prospective, multipathogen studies. Foundation of Key Laboratory of Veterinary Biotechnology, National Key Research and Development Program of China, and the International Joint Laboratory on Tropical Diseases Control in Greater Mekong Subregion.
Traditional fermented foods represent complex microbial ecosystems shaped by substrate composition, indigenous processing practices, and local environmental conditions. However, the microbiomes of many traditional fermented foods from Northeast India remain poorly characterized. In this study, the bacterial communities associated with thirteen traditional fermented foods from Mizoram, India, representing both plant- and animal-derived fermentations, were characterized using full-length 16S rRNA gene sequencing on the Oxford Nanopore platform. Sequencing generated approximately 1.94 million high-quality reads, enabling high-resolution taxonomic profiling of the fermented food microbiomes. The microbial communities were predominantly composed of Bacillota, Pseudomonadota, and Cyanobacteriota, although their relative abundances varied considerably among fermented food types. Plant-derived fermented foods were enriched with fermentative bacterial genera, including Bacillus, Lactobacillus, Lacticaseibacillus, Pediococcus, and Weissella, whereas animal-derived fermented foods showed a greater abundance of anaerobic taxa, particularly Clostridium. Alpha diversity analyses demonstrated higher microbial richness in plant-derived fermented foods, while beta diversity revealed clear substrate-dependent clustering of microbial communities. Functional prediction indicated that metabolism-related pathways, particularly carbohydrate and amino acid metabolism, predominated across the fermented food microbiomes, and LEfSe analysis identified distinct microbial biomarkers associated with plant- and animal-derived fermentations. Collectively, these findings provide the first comprehensive microbiome characterization of traditional fermented foods from Mizoram and demonstrate that fermentation substrate is a major determinant of microbial community composition and predicted functional potential. Although full-length 16S rRNA gene sequencing improved taxonomic resolution, taxonomic interpretations were made cautiously at the genus level where appropriate because of the inherent limitations of 16S rRNA gene-based classification.
The growing food allergy epidemic is thought to be related to changing environmental factors, particularly changes in the gut microbiome. While prior work has demonstrated that food allergy can be modulated by gut microbes, little is known about how food allergen-specific CD4+ T cells are affected by gut microbial composition. Here, we report that food allergy severity differs between mice obtained from 2 different specific pathogen-free mouse vendors (Jackson Labs [Jax] and Taconic Biosciences [Tac]). Mice from Tac develop diarrhea and anaphylaxis after fewer allergen exposures than mice from Jax. Using food allergen peptide: MHCII tetramers, we also find that Tac mice have fewer allergen-specific regulatory T cells in the small intestine compared to mice from Jax with concomitant increase in allergen-specific Th2 cells. In addition, Tac mice have increased intestinal permeability. Increased food allergy severity, phenotype of allergen-specific T cells, and increased gut permeability were transferable to Jax animals via co-housing, which corresponded to a shift in Jax microbial communities towards those found in Tac mice. Our findings demonstrate that food allergen-specific Treg cells can be modulated by gut microbial community composition, which in turn is correlated to food allergy severity.
This study presents a novel hybrid biosensor integrating chicken IgY antibodies and quantum dot-labeled DNA aptamers for rapid, sensitive detection of staphylococcal enterotoxin A (SEA) in food samples. The system leverages the unique advantages of avian IgY, which avoids false positives from staphylococcal protein A interference, combined with high-affinity aptamers (K d= 84 nM) selected through magnetic bead-SELEX for specific detection. The QD-conjugated aptamer reporter system enables visual fluorescence readout with a detection limit of 1 ng/mL within 45 min, offering a faster workflow, operational simplicity, and an estimated consumable cost of <$1 per test under bulk-production conditions. Validation studies using spiked and field-collected food matrices demonstrated strong agreement with PCR-based analysis, including complete concordance in the 50 tested food samples, supporting the platform's reliability for food safety monitoring. This biosensor represents a significant advancement in point-of-need toxin detection, offering laboratory-grade accuracy in a field-deployable format that requires no specialized equipment, making it particularly valuable for resource-limited settings and routine food safety screening programs.
Artificial light at night (ALAN) is an increasingly pervasive global change driver, yet its effects on the interconnected biogeochemical cycling of soil carbon (C) and nitrogen (N) remain poorly understood. Here, we conducted an in situ dual-isotope (13C-CO2 pulse labelling and 15N addition) experiment combined with multitrophic microbial network analysis in a semi-arid meadow steppe to explore the impact of ALAN on belowground C and N cycling and the underlying mechanisms. We found that ALAN significantly accelerated the turnover of newly fixed plant C, a process primarily fueled by the increased root-derived C input from ALAN-stimulated plant biomass (the bottom-up C-pump). Concurrently, while this plant-driven C surplus primarily fueled elevated soil respiration, ALAN substantially enhanced microbial 15N retention, resulting in a pronounced asymmetric biological allocation of assimilated C and N within the biotic pools. This stoichiometric divergence was underpinned by a physiological shift toward a conservative microbial N retention strategy, whereby soil microorganisms increased N retention efficiency to counterbalance the stoichiometric C surpluses induced by enhanced plant inputs. Crucially, we found that this enhanced biological N retention was predicted not by microbial taxonomic diversity but was closely associated with a directional trend toward reduced micro-food web structural connectivity. This fragmented network architecture potentially altered top-down trophic controls, which compounded the plant-driven stoichiometric imbalance by limiting predator-mediated N mineralization and thereby promoting relative N retention within the microbial biomass. Our findings highlight light pollution as an underappreciated global change driver that specifically accelerates plant-mediated bottom-up C loss while simultaneously promoting microbial N retention via fragmented micro-food webs in increasingly illuminated ecosystems.
Strains of food-derived microbes can become facultative pathogens in susceptible human hosts. Surprisingly, we previously isolated Debaryomyces hansenii, a yeast common in fermented foods, from Crohn disease (CD) ulcers, raising questions about its strain-specific traits that influence host interactions. Here, we further developed the genetic tractability of D. hansenii and identified a single adhesin, Hil1, as a major determinant of colony morphology, biofilm formation, and immune targeting in CD patients. We used Agrobacterium tumefaciens-mediated transformation to perform a forward genetic screen in a food-derived reference strain. We isolated mutants that converted from a wrinkled, biofilm-forming phenotype to a smooth, non-adherent phenotype characteristic of CD patient isolates. Mapping of multiple insertion sites showed a disrupted subtelomeric Hyr/Iff-like adhesin gene, herein referred to as HIL1. CRISPR-Cas9-mediated deletion of HIL1 recapitulated the mutant phenotype, demonstrating that HIL1 was necessary for biofilm formation and high cell-surface hydrophobicity phenotypes. To contextualize these findings, we performed comparative genomics on a D. hansenii strain collection to assess allelic variation in the number of HIL1 tandem repeats. Longer alleles in food strains correlated with increased biofilm formation, while CD-isolated strains contained shorter HIL1 alleles and reduced binding to surfaces. Serology profiling showed that HIL1 was a direct antigenic target of circulating immunoglobulin G (IgG) in CD patients. Together, these results suggest Hil1 is a key, strain-variable adhesin shaping fungal surface properties and host immune recognition. This work establishes D. hansenii as a genetically tractable system and shows how adhesin polymorphisms may influence fungal behavior in food and disease contexts. Debaryomyces hansenii is a yeast that is common in food and is generally recognized as safe for human consumption, though recently it has been identified within diseased regions of the intestine in Crohn disease patients. A current need is to determine the genetic and phenotypic differences between safe food isolates and isolates from human Crohn disease patient ulcers. Here, we used a loss-of-function genetic screen and identified HIL1, an adhesin that we found mediates cellular adhesion in many food strains but not in patient strains. We identified circulating HIL1-reactive antibodies in patients with Crohn disease, indicating that food strains can be a target of host immune responses through Hil1.
The application of next-generation sequencing (NGS) is rapidly expanding for antimicrobial resistance (AMR) surveillance and clinical decision-making. However, despite a high AMR burden, many low- and middle-income countries lack the infrastructure and technical capacity required to implement sequencing-based approaches. To address this gap and demonstrate the practical application of the technology, we compared the performance of nanopore sequencing combined with real-time analysis using the EPI2ME platform in a mobile laboratory with that of a hybrid sequencing approach integrating Illumina short reads and nanopore long reads. In this exploratory diagnostic evaluation study, 25 Escherichia coli isolates obtained through the German annual national AMR monitoring program were included. Isolates resistant to at least one β-lactam antibiotic were considered as resistant isolate. Genomic DNA was subjected to both Illumina and nanopore sequencing. Following the sequencing, the EPI2ME whole genome sequencing pipeline was used to detect and identify β-lactamase genes in E. coli isolates. As the comparator, both long and short read-based hybrid assemblies were used to detect and identify the β-lactamase genes. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the sequencing approaches were calculated against the phenotypic classification of E. coli isolates, treating the presence or absence of β-lactamase-encoding genes as the genotypic predictor of β-lactam resistance. A total of 36 β-lactamase-encoding genes, predominantly from the bla CTX-M, bla SHV, and bla TEM families, were detected. Both analytical approaches achieved a promising accuracy towards predicting β-lactam resistance. Moderate agreement was observed between methods, with an overall concordance of 68.9%. A distinct distribution pattern of β-lactamase genes was observed across the MIC ranges of the isolates for third-generation cephalosporins and the carbapenems. These findings support the feasibility of deploying nanopore sequencing in mobile suitcase laboratories to strengthen AMR surveillance in food production systems. Given its portability, rapid turnaround time, and minimal infrastructure requirements, the approach may also have broader applications for One Health based AMR surveillance, clinical decision-making, and outbreak investigations in resource-limited settings.
Weissella cibaria, a heterofermentative lactic acid bacterium, is recognized for its antimicrobial activity against a range of pathogenic microorganisms. Therefore, the objective of this study was to evaluate the antimicrobial potential of W. cibaria strains, with an emphasis on their organic acids, against target microorganisms commonly associated with milk and dairy products (Listeria monocytogenes, Escherichia coli, Staphylococcus aureus, and Salmonella). To this end, this study evaluated the minimum inhibitory concentration (MIC) of cell-free supernatants (CFS) from three W. cibaria strains (W21, W25, and W42) and their 1:1:1 mixture. Furthermore, the CFS mixture was used in a cheese-simulating matrix to inhibit the target microorganisms. Using HPLC, the organic acids present in the CFS were identified and quantified. Finally, combinations of identified synthetic organic acids were tested for effective inhibition of the target microorganisms. The CFS from strains W21 and W25 and the CFS mixture exhibited better MIC, while a higher concentration of the CFS from W42 was required to inhibit the tested strains. In a cheese-simulating matrix, the CFS mixture significantly reduced the survival rate of the target microorganisms (p < 0.05). By HPLC, the organic acids detected were lactic, acetic, isovaleric, propionic, butyric, valeric, and malic acids. Strain W42 stood out for the absence of isovaleric and valeric acids, while strain W25 stood out for the absence of malic acid. A total of 127 combinations of synthetic organic acids were formulated. L. monocytogenes and Salmonella were predominantly inhibited by combinations of three acids, while E. coli and S. aureus were predominantly inhibited by combinations of 4 or 5 acids and 3 or 5 acids, respectively. The predominance of lactic, acetic, and isovaleric acids was associated with inhibition of the tested microorganisms, while valeric acid was not required for the inhibitory effect of the most effective combinations. These findings support the use of W. cibaria CFS as a potential natural antimicrobial strategy in dairy products and indicate that organic acid combinations may contribute to the inhibition of food-associated microorganisms.
Pregnant women are at higher risk of contracting listeriosis, which can lead to serious perinatal complications. This study evaluated associations between Listeria monocytogenes exposure, infant perinatal outcomes, and hospital resource use in a cohort of 1604 Australian mother-infant dyads. Maternal L. monocytogenes exposure was estimated from self-reported intake of foods from a validated food frequency questionnaire that potentially harbor L. monocytogenes. Infant outcomes obtained from hospital medical records included birth mode, preterm birth, birthweight, and admission to special care nursery (SCN)/neonatal intensive care unit (NICU). Infant hospital resource use was measured by infant length of stay (LOS), LOS in SCN/NICU, and days excluding SCN/NICU. Multinomial, negative binomial, and hurdle models, were performed to examine Listeria Food Exposure Score (LFES) associations with infant outcomes, total LOS, and LOS excluding and within SCN/NICU. All models were adjusted for covariates including smoking, parity, maternal age, BMI, and SEIFA IRSAD decile. Mean (SD) maternal age was 32.0 (5.0) years, and median (IQR) gestation was 39.0 (38.1, 40.0) weeks. Although adjusted results showed a statistically significant association between LFES and reduced infant LOS excluding SCN/NICU (β = 0.99; 95% CI: 0.979, 0.998, p < 0.03), the effect size was minimal, with minor clinical significance. There were no significant associations with infant birth mode, preterm birth, low birthweight, size for gestational age, macrosomia, admission to NICU/SCN, total LOS in hospital, and SCN/NICU (all p > 0.05). Future research should explore these associations among ethnically diverse women at earlier stage of pregnancy and include the assessment of food safety practices in the analyses.
Increased intestinal permeability and proinflammatory status have been linked with increased risk of autoimmune diseases such as type 1 diabetes. The contribution of dietary composition to gut maturation in infants consuming complementary foods remains unclear. The aim was to assess whether intake of different nutrients is associated with the level of intestinal permeability and gut inflammation markers of infants aged 6-12 months. This was a secondary analysis using existing cohort data from a randomized controlled trial, the Early Dietary Intervention and Later Signs of Beta-Cell Autoimmunity (EDIA) Study (N = 73). Weighed 3-day food records were collected at 6, 9, and 12 months of age. At the same age points, lactulose/mannitol tests (N = 72) were performed for assessment of intestinal permeability, and stool samples (N = 72) collected for analysis of gut inflammation markers calprotectin and human β-defensin-2 (HBD-2). Generalized estimation equations (GEE) were used to assess associations between repeated measures of energy-adjusted nutrient intakes with intestinal permeability, calprotectin, and HBD-2. Higher intake of saturated fat was associated with higher intestinal permeability, calprotectin, and HBD-2, while higher intake of dietary fibre, pyridoxine (vitamin B6), iron, magnesium, and potassium were associated with lower permeability, calprotectin, and HBD-2. The nutrients with inverse associations were intercorrelated with each other. Additionally, the intercorrelated nutrients correlated negatively with the proportion of total energy intake from breast milk, which was associated with higher permeability and inflammation markers. Adjusting models for the proportion of energy from breast milk attenuated many of the associations of nutrients, especially with permeability, but the intake of dietary fibre remained associated with lower calprotectin and HBD-2, and intake of potassium with lower calprotectin and permeability. While breastfeeding remains a major determinant of gut maturation, consumption of complementary foods higher in intercorrelated nutrients including dietary fibre and potassium is associated with lower gut inflammation markers and, less robustly, reduced intestinal permeability in infants with genetic susceptibility to type 1 diabetes aged 6-12 months. Trial registration clinicaltrials.gov NCT01735123.
Meat adulteration is a significant global food safety challenge, creating a pressing need for rapid and on-site detection technologies. Herein, we present an intelligent one-pot biosensing platform termed one-pot TLAMP-PfAgo assay (OTPA) that integrates the rapid amplification of turn-back loop primer-accelerated loop-mediated isothermal amplification (LAMP) (TLAMP) with the sequence-specific detection of Pyrococcus furiosus Argonaute (PfAgo). This system features a clever heat-activatable design using microcrystalline wax to spatially separate reactions within a single tube, enabling contamination-free and streamlined operation. The OTPA assay achieves sensitive and specific detection, with limits of detection as low as 3×10-4 ng/μL for pork DNA and 2×10-4 ng/μL for beef DNA within 30 min. It successfully enables duplex target identification and has been validated with commercial meat products, showing perfect concordance with standard polymerase chain reaction (PCR)-based qualitative detection. Notably, the result can be directly visualized under blue light, underscoring the substantial potential of this cost-effective and simple platform for point-of-care testing (POCT) and intelligent biosensing in food safety surveillance. 肉类掺假是全球食品安全面临的一项重大挑战，因而迫切需要开发快速的现场检测技术。在此，本研究提出了一种名为OTPA（一锅法TLAMP-PfAgo分析）的智能一锅法生物传感平台。该平台将基于折返环引物加速的环介导等温扩增技术（LAMP）（TLAMP）与激烈火球菌（Pyrococcus furiosus）的Argonaute蛋白（PfAgo）的序列特异性检测功能完美融合，采用一种巧妙的热激活设计，利用微晶蜡在单管内实现反应的空间隔离，从而确保了操作的无污染与流程的简化。OTPA分析法具备极高的灵敏度与特异性，在30分钟内对猪肉DNA和牛肉DNA的检测限分别低至3×10-4 ng/μL和2×10-4 ng/μL。此外，它成功实现了双靶标鉴定，并在市售肉制品中得到了验证，其结果与标准的PCR定性检测高度一致。值得注意的是，检测结果可在蓝光下直接通过肉眼可视化，这充分凸显了这种经济高效且简便的平台在食品安全监测的即时检测（POCT）与智能生物传感领域中所具备的巨大潜力。.
Chitosan-Spent coffee ground-ZnO (Chi-SCG-ZnO) biobased film has been successfully synthesized and evaluated for its potential application as an active food packaging. The biobased film was prepared via a solution casting method and characterized in terms of mechanical properties, water vapor permeability (WVP), water contact angle (WCA), UV-blocking ability, antibacterial activity, biodegradability, and environmental toxicity. The incorporation of SCG and ZnO improved the physicochemical and functional properties of the film. The tensile strength increases from 5.74 MPa (Chi) to 10.98 MPa (Chi-SCG-ZnO). The Chi-SCG-ZnO film exhibited lower WVP (0.52 g.m-2.h-2) compared with Chi (0.62 g.m-2.h-2), indicating improved moisture barrier properties. The surface hydrophobicity also increased, as shown by a higher WCA (82.98o) compared with Chi (64.25o). In addition, Chi-SCG-ZnO demonstrated excellent UV-blocking ability, reaching 99.74% in the UVB region and 92% in the UVA region. The film also exhibited stronger antibacterial activity against Escherichia coli (9 mm) and Staphylococcus aureus (10 mm) compared with Chi. Application in grape packaging showed that Chi-SCG-ZnO maintained better fruit quality during storage than Chi. The biodegradability test indicated higher degradation of Chi-SCG-ZnO compared with Chi. These results demonstrate that Chi-SCG-ZnO has promising potential as an active and biodegradable food packaging material.
Antimicrobial resistance is a growing global public health concern, and carbapenem-resistant Enterobacterales (CRE) represent one of the most urgent threats due to limited treatment options. While CRE are primarily associated with healthcare settings, increasing evidence suggests that food systems may serve as additional reservoirs and transmission interfaces for clinically significant antibiotic-resistant bacteria. This study aimed to characterize CRE from vegetables, with a focus on isolates carrying carbapenem resistance genes on mobile genetic elements and evaluating their transferability to a recipient bacterium. Commercial vegetable samples were collected from retail grocery stores or farmers' markets and screened for CRE using selective enrichment and CHROMagar™ mSuperCARBA™. Antimicrobial susceptibility testing was performed according to Clinical and Laboratory Standards Institute (CLSI) guidelines. Whole-genome sequencing using Illumina short-read and Oxford Nanopore long-read platforms was conducted to characterize resistance genes and the plasmid sequence. Conjugation assays were performed to assess plasmid transferability to Escherichia coli DH5α. The results indicated that a carbapenem-resistant Kluyvera sichuanensis isolate (CA-CRE-C23) was recovered from a spinach sample. Genomic analysis identified a 51,477-bp hybrid ColKP3-IncX3 plasmid carrying the carbapenemase gene blaOXA-181 and quinolone resistance gene qnrS1. Conjugation experiments, disk diffusion, and sequencing confirmed successful plasmid transfer from K. sichuanensis isolate (CA-CRE-C23) to E. coli DH5α. The plasmid structure closely resembled globally disseminated IncX3 hybrid plasmids carrying blaOXA-181, which have been predominantly reported in clinical isolates. In conclusion, the detection of a transferable hybrid plasmid harboring blaOXA-181 in a fresh produce-associated K. sichuanensis isolate expands the known host range of this resistance gene and underscores the importance of food-chain surveillance in understanding the environmental dissemination of high-risk antimicrobial resistance genes.
Salmonella enterica is a major foodborne pathogen associated with beef contamination and increasing antimicrobial resistance, particularly in low- and middle-income countries. This study investigated the prevalence, antimicrobial resistance patterns, and genetic diversity of S. enterica isolated from retail beef samples in Peshawar, Pakistan, using combined phenotypic and molecular approaches. A total of 250 beef samples were collected from urban, rural, and peri-urban retail markets. Salmonella enterica was isolated using standard culture and biochemical methods and confirmed by PCR targeting the invA gene. Antimicrobial susceptibility was determined by the Kirby-Bauer disk diffusion method, and genetic diversity was assessed using randomly amplified polymorphic DNA PCR (RAPD-PCR). Overall, S. enterica was detected in 150 samples (60.0%), with higher prevalence in rural markets (77.5%) compared with urban (52.5%) and peri-urban (50.0%) locations. A high percentage of isolates were resistant to azithromycin (94.0%), tetracycline (68.7%), and streptomycin (52.0%), indicating widespread multidrug resistance among isolates. Five isolates (3.3%) were identified as extended-spectrum β-lactamase (ESBL) producers and carried blaTEM-1, blaCTX-M-15, or blaSHV-12 genes. RAPD-PCR analysis revealed substantial genetic heterogeneity, with isolates distributed across multiple phylogenetic clusters, suggesting diverse contamination sources rather than clonal dissemination. These findings indicate that retail beef in Peshawar represents a significant reservoir of genetically diverse and antimicrobial-resistant S. enterica. The combined use of phenotypic methods with targeted molecular and low-cost genotyping approaches provides a practical framework for surveillance of foodborne Salmonella in resource-limited settings.
Reproductive microbiota is known to modulate host immunity, reproductive physiology, and longevity, yet their interactions with host genetics, sex, and mating status remain relatively underexplored in most insects. Some studies suggest that species (and even populations) can vary in the microbial communities associated with their reproductive tissues. Sexually transmitted microbes can also directly influence population growth and reproductive fitness, making them relevant even for applied insect management strategies. The economically relevant, Black Soldier Fly (BSF; Hermetia illucens), is known for its applications in organic waste management and sustainable food production. Despite several papers on the larval gut microbiota, there is no study (to date) that explores the reproductive microbiota of adults. This study characterizes the reproductive microbial communities of eight genetically distinct BSF populations using 16S rRNA gene sequencing. The results revealed striking, sex-specific microbial signatures within reproductive tissues, dynamically shaped by mating. Virgin females harboured diverse and functionally intriguing taxa, including Staphylococcus, Brevibacterium, and Corynebacterium, which likely supported ovarian maturation and reproductive readiness. Remarkably, mating triggered a dramatic shift, with unclassified Enterococcaceae emerging as the dominant taxon in mated females, suggesting selective microbial retention or transmission with potential implications for post-mating longevity. In contrast, although mated males showed enrichment of Rhodococcus_C_375578, their overall microbiota composition remained comparatively stable pre- and post-mating. Most notably, the genetic lineage significantly influenced the magnitude of sex- and mating-related bacterial shifts in reproductive microbiota. These novel findings advance the understanding of the complex dynamics of reproductive microbiota in insects and provide a foundation for optimizing breeding strategies and colony management in commercial BSF production systems.
Biofilms formed at air-liquid interfaces represent a significant challenge for the food industry due to their resistance to cleaning and the difficulty of detection. This study systematically investigates the ability of five bacterial species (E. coli, B. subtilis, C. sakazakii, P. fluorescens and L. plantarum) to form interfacial biofilms on stainless steel AISI 304, stainless steel AISI 316, and aluminium. Biofilm formation was confirmed by crystal violet staining and quantified via viable cell count. C. sakazakii produced the highest biofilm biomass (>6 log CFU cm-2 on stainless steel, in milk), whereas P. fluorescens (minimal media) and L. plantarum (milk) yielded no viable cells, suggesting limitations in recovery or altered physiological states. Spectral imaging techniques (FT-IR, Raman, SWIR, VNIR) were used to analyse biofilm formation. Results demonstrated a strong influence of surface type and growth medium on spectral signatures and classification performance, with stainless steel substrates supporting higher bacterial growth than aluminium under nutrient-rich conditions. Principal Component Analysis (PCA) showed clear separation of contaminated samples in milk, while broth samples were more stable but less distinct. All techniques detected contamination, with SWIR and VNIR providing more consistent classification performance, particularly when combined with sub-regional analysis and data fusion. Broth-incubated biofilms were more reliably classified than those in milk, likely due to reduced spectral complexity. The variability across test sets underscores the need for careful sample selection and robust model validation. Overall, spectral imaging is a promising non-destructive approach for biofilm detection, with SWIR and VNIR showing the best performance.
The One-HornedRhino (Rhinoceros unicornis) is a vulnerable species that inhabits the lowlands of Nepal. With the existence of several threatening factors, such as poaching, natural disasters, food toxicity, and diseases, their survival and overall health are continuously threatened. In this context, diseases caused by intestinal parasites can serve as a warning sign. However, the occurrence of parasite types and the predicted impacts of parasitism in these megaherbivores remained unclear and questionable in Nepal. The current study aimed to determine the presence of endoparasites (protozoa and helminths) inhabiting these hosts, which naturally live freely outside the Chitwan National Park (CNP) in Central Nepal. Sixty fresh fecal samples (N = 60) from five different sampling locations along the nearby territories were collected opportunistically using noninvasive methods and transported to the research laboratory. Coproscopy was carried out following standard protocols and laboratory techniques, including direct wet mount, concentration, and acid-fast staining methods. The results showed a 100% prevalence rate, with 16 species of intestinal parasites (Protozoa: 5; Helminths: 11). Furthermore, the chi-square (χ 2) goodness-of-fit test indicated a statistical difference between the frequencies of protozoan (16.7%) and helminth (98.3%) detections (p < 0.05, χ 2 = 5.04, degree of freedom = 1). Entamoeba sp. and Eimeria spp. were the most frequently recorded protozoa, while Anoplocephala spp. and Strongyle were the most frequently recorded helminths. Additionally, monoparasitic infection was not observed, and quadruplet infection (coinfection with four parasite species) represented the most prevalent pattern (25%). This is the first study to document protozoan and helminth diversity and multiparasitism in free-ranging rhinos outside protected areas in Nepal. Our findings of 100% prevalence and 16 diverse parasite taxa highlight the need for urgent antiparasitic drug treatment for those rhino populations.
Restaurant wastewater (RWW) represents an abundant yet largely unexplored nutrient source for enriching ureolytic microbial consortia applicable to microbial-induced calcite precipitation (MICP). This study characterised RWW collected from a food-service establishment in Johor, Malaysia (COD 1,341 mg/L; BOD 837 mg/L; pH 6.8), and systematically evaluated its capacity to support indigenous ureolytic bacterial enrichment across three media formulations: yeast extract-based (Medium-1), nutrient broth-based (Medium-2), and brown sugar-based (Medium-3). Medium-1 delivered the strongest performance, achieving OD600 = 1.29 ± 0.06, urease activity = 17.42 ± 1.19 mM urea hydrolysed min- 1, and CaCO3 precipitation = 2.81 ± 0.17 g/L. Optimal bioactivity was recorded at pH 8 and 30 °C, conditions closely aligned with the tropical collection environment. 16 S rRNA amplicon sequencing (DADA2 pipeline; SILVA nr v138.1) yielded 101,869 quality-filtered reads across 116 amplicon sequence variants (ASVs; Shannon H = 2.79), identifying a co-dominant community of Firmicutes (50.83%) and Proteobacteria (48.36%), with Sporosarcina (6.09%), Bacillus (3.35%), Lysinibacillus (2.63%), and Raoultella (34.39%) as principal ureolytic contributors. Soil biocementation trials returned a mean surface strength of 423.3 ± 21.6 psi and a CaCO3 content of 16.64 ± 1.72%. Heavy metal immobilisation efficiencies reached 99.60% for Cd2+, 81.87% for Ni2+, 42.47% for Cr3+, and 22.47% for Cu2+ at 10 mg/L. XRD, FTIR, TGA, and DSC collectively confirmed a thermally stable, mineralogically pure biogenic calcite (> 96.7% residue at 894 °C). Collectively, these findings establish RWW-enriched consortia as functionally capable, cost-effective biocatalysts for sustainable MICP, in support of circular economy objectives within tropical urban contexts.
This study aimed to investigate the anti-obesity properties of Komagataeibacter rhaeticus SLAM-JS1B derived metabolites, a bacterial strain isolated from kombucha. The effects were assessed in mice with obesity induced by a high-fat diet. Supplementation with K. rhaeticus SLAM-JS1B derived metabolites significantly attenuated body weight gain without altering food intake. Serum total cholesterol, triglyceride, and low-density lipoprotein levels were significantly reduced, together with improved indicators of hepatic damage. In addition, hepatic steatosis and adipose tissue accumulation were markedly attenuated. These metabolic improvements were linked to lower hepatic expression of genes related to lipogenesis and cholesterol production. In the colon, supplementation with K. rhaeticus SLAM-JS1B derived metabolites increased the expression of genes related to intestinal barrier integrity and lowered the expression of pro-inflammatory cytokine genes. Fecal metabolomic analysis further revealed increased fecal cholesterol excretion following K. rhaeticus SLAM-JS1B derived metabolites supplementation. Moreover, gut microbial composition was altered in a manner consistent with improved metabolic status. Collectively, these findings suggest that K. rhaeticus SLAM-JS1B derived metabolites may represent a promising dietary strategy for the prevention or management of obesity and related metabolic disorders, particularly in contexts where the use of live microorganisms is undesirable.
Fermentation microbiomes play essential roles in food production, feed preservation, waste valorization, and diverse sustainable industrial processes. Although multi-omics and systems biology have substantially advanced our understanding of their assembly, interactions, and functional dynamics, industrial translation remains constrained by fragmented datasets, limited causal validation, and transport constraints during scale-up. Large language models act as upper-level knowledge and workflow orchestrators, accelerating data integration, hybrid AI-mechanistic modeling, hypothesis generation, and perturbation-guided learning. Collectively, these advances enable fermentation microbiome research to move beyond descriptive omics toward mechanism-guided synthetic microbial community design, causal validation, and scalable biomanufacturing.