Journal of Dairy Science

The emergence of Highly Pathogenic Avian Influenza (HPAI 2.3.4.4b) in dairy herds in 2024 across 19 states in the United States of America has raised concerns regarding the potential national and global zoonotic impact. All recent modeling efforts implemented homogenous cattle-to-cattle (both intra and inter-herd) transmission models which did not capture the real-world heterogeneity in mixing of animals, individual variations in susceptibility and infectiousness and clinical incidences across pens and lactation groups. The aim of this study was to develop a heterogenous transmission model to estimate the epidemiological parameters for intra-herd HPAI transmission on Californian dairies. We developed a validated stochastic agent-based model to estimate the epidemiological parameters for intra-herd HPAI transmission on California dairies. The hierarchical agent-based model also parameterized stochastic cattle movements within-herd to simulate real dairy management practices. A cow-level SEIR transmission approach was assumed during the outbreak. A novel Bayesian Optimizer with Gaussian Process (BO-GP) was fitted to the agent-based model for validation which converged within 25-40 iterations (out of 100 per farm) with minimal loss over two distinct error metrics, namely, Poisson loss function and temporal distance metric. Our optimized simulations estimated an average R0 was around 10.7-10.8 across all farms within the first 15 days of observed outbreak on four dairy farms with a mean effective transmission rate of 4.5% per contact between susceptible and infectious cows within each pen. Our model demonstrated that the movement of cows between pens ensured localized clusters of outbreaks within the sub-herds (pen population) that prolonged the overall outbreak within farms. We estimated the total duration of infection between 14.5 and 28 days, which is higher than the estimates from the homogenous models. With an integrated hierarchical agent-based model combined with Bayesian approximation, we produced actionable insights on the epidemiology of intra-farm spread of HPAI within cow herds, thereby guiding both future model development and applied disease control strategy.

The growth and performance of cattle is dependent on efficient feed conversion, facilitated by the gut microbiome, including both prokaryotic and eukaryotic organisms. Extrinsic factors such as management practices and intrinsic factors like age and genetics can shape these communities. To characterize the relative effects of such factors on the prokaryotic and eukaryotic microbiomes, fresh fecal samples from dairy heifers (n = 8), beef heifers (n = 8), bred beef heifers (n = 8), and adult beef cows (n = 8) were subjected to paired 16S rRNA (V4) and a novel 18S rRNA (V4-V5) metabarcoding assay, and 18S rRNA amplicons were annotated using two databases in parallel. Taxonomic resolution of 18S rRNA amplicons was excellent, with SILVA and EUKARYOME databases each providing better resolution of different groups of organisms. While no differences in eukaryotic richness or alpha diversity were detected, beta diversity differed in all pairwise comparisons except between beef heifers and bred beef heifers. Prokaryotic richness was lower in adult beef cows than the other groups, and beta diversity differed in all pairwise comparisons. Adult beef cows were the most distinct, characterized by replacement of fungal organisms with protozoa and loss of thermophilic bacteria in the eukaryotic and prokaryotic compartments, respectively. Among numerous differentially abundant features, dairy heifers harbored greater abundance of Cooperia oncophora while beef heifers contained greater abundance of Buxtonella sulcata and Mucor. Correlation network analysis revealed multiple significant positive and negative associations between bacterial, fungal, and protozoal features during maturation of the fecal microbiome. Collectively, these data highlight the richness and diversity of both the prokaryotic and eukaryotic compartments of the bovine fecal microbiome, and the influence of management practices and age on both communities. From a technical standpoint, these data demonstrate the ability to resolve fungal, protozoal, and metazoan organisms, including potential parasites and pathogens, with a single metabarcoding assay. They also highlight the nuanced differences between databases and the value of parallel analyses using more than one database.

The study of animal personality has gained increasing attention in recent years, as researchers recognize that non-human animals exhibit stable individual differences in behaviour. In dairy cattle, understanding personality traits holds potential for improving animal welfare, enhancing management practices, and supporting productivity outcomes. However, validating the behavioural tests used to assess these traits remains a significant challenge. The present study aimed to evaluate the consistency and validity of five commonly used behavioural tests in dairy cows: the Human Approach Test (HAT), Sudden Test (SUD), Novel Object Test (NOT), Open Field Test (OFT), and Novel Surface Test (NST). These tests were chosen for their potential to assess personality dimensions that parallel components of human personality, particularly Openness, Neuroticism, Extraversion, and Agreeableness. Repeated assessments were conducted across three time periods to evaluate both intra-test (temporal) and inter-test (contextual) consistency. Results revealed that only the SUD exhibited significant behavioural consistency across all periods, suggesting its potential as a reliable tool for personality assessment. Additionally, a moderate correlation between the HAT and SUD was observed, indicating that these tests may capture overlapping personality traits. In contrast, the remaining tests (NOT, OFT, NST) lacked consistent behavioural patterns over time and across contexts, raising questions about their reliability and the complexity of trait expression. These findings highlight the challenges of identifying stable personality traits in cattle and emphasize the need for more rigorous validation of behavioural tests. This research contributes to the growing field of animal personality by providing insights into trait stability, the limitations of current testing methods, and the potential application of personality assessment in future livestock management practices.

Macrophages, alongside mammary epithelial cells, are major contributors to the innate immune response triggered by bacterial invasion of the mammary gland. To investigate breed-specific immune mechanisms, we stimulated monocyte-derived macrophages from Holstein and Normande cattle with Escherichia coli and Streptococcus uberis, two major mastitis pathogens. We monitored bacterial survival using [methode] and profiled gene expression via RNA-seq. E. coli exhibited significantly higher survival 6 hours post-infection compared to S. uberis (p < 0.05). Moreover, E. coli induced a robust transcriptional response, with 4403 differentially expressed genes (FDR < 0.05, |log2FC| > 1), versus only 813 for S. uberis. Qualitative differences were also observed: genes such as CCL2 and CSF1 were uniquely induced by S. uberis. Additionally, macrophages from Holstein and Normande cattle showed distinct expression profiles: 185, 45, and 174 genes were differentially expressed at steady state or after E. coli or S. uberis stimulation, respectively. These genes were enriched in immune-related pathways. Overall, our results demonstrate a stronger response to E. coli, although S. uberis specifically induces a subset of genes. The observed breed-specific differences may reflect underlying immune mechanisms that could influence disease susceptibility, warranting further functional validation.

The gut-airway axis is increasingly recognised as an important determinant of respiratory health in livestock. However, the interactions among respiratory infections, microbial communities, mucosal barrier function, and microbial metabolism that underpin this axis remain poorly understood. Here, we combined longitudinal immunological, mucosal, microbiological, and metabolic analyses to characterise host-microbiota interactions during antibiotic treatment and experimental bovine respiratory disease (BRD). We found that the bovine respiratory tract is organised as a hierarchically structured mucosal ecosystem, with compartment-specific mucin expression patterns associated with distinct microbial communities. Within this framework, increasing respiratory pathogen loads were associated with greater clinical severity, oxidative stress, acute inflammation, and targeted alterations in mucosal regulation and microbial composition, while serum amyloid A emerged as the host biomarker most closely reflecting infection intensity. Although infection altered host and microbial states, antimicrobial treatment represented the dominant ecological perturbation. Antibiotic exposure reshaped both respiratory and intestinal microbial communities, reducing diversity, increasing community drift, and driving coordinated ecological transitions across the gut-airway axis that exceeded the effects of infection alone. Importantly, these compositional changes were accompanied by profound reductions in acetate, propionate, butyrate, and valerate, identifying short-chain fatty acids as a major functional consequence of gut microbiota disruption. Integrative multi-omics analyses revealed that mucosal organisation, microbial diversity, microbial metabolism, and immune activity were embedded within a highly structured gut-airway network. Together, these findings support a holobiont view of BRD in which disease outcomes emerge not only from pathogen-host interactions but also from ecological disturbances that propagate across interconnected mucosal compartments. Notably, antimicrobial treatment affected ecosystem structure and function more than infection itself, highlighting potential trade-offs between pathogen control and microbiota stability. Preserving microbial resilience and its metabolic functions may therefore represent an important component of future strategies to improve respiratory health and disease resilience in cattle.

Abstract Napier grass (Cenchrus purpureus, NG) is an important forage in tropical and subtropical livestock systems, but the extent of naturally occurring variation in its methane (CH4) production and its potential for selecting low-CH4 emitting genotypes remains poorly characterised. This study evaluated variation in CH4 yield among NG accessions, examined the effects of season and water stress, validated contrasting CH4 phenotypes across donor cows, and identified elite accessions combining low CH4 yield with favourable digestibility. A total of 750 NG samples representing 84 accessions grown under dry and wet seasons and moderate and severe water-stress were evaluated using a high-throughput in-vitro fermentation system. To further identify forages combining low-CH4 production with acceptable digestibility, a multi-trait selection approach was applied using biologically defined thresholds for CH4 yield and in vitro organic matter digestibility (IVOMD), followed by a weighted index assigning 60% weight to CH4 yield and 40% to IVOMD. Methane yield varied widely among accessions, ranging from 0.02 to 5.07 mL/g DM and was affected by growing conditions. Methane production was generally lower during the wet season, while water stress was associated with changes in forage quality and fermentation characteristics. Low-CH4 producing accessions generally had lower IVOMD, crude protein and metabolizable energy and relatively higher fibre than high-CH4 producing accessions. Accessions initially classified as low and high-CH4 emitters based solely on CH4 yield were subsequently validated using rumen inoculum from three individual donor cows. Although the magnitude of CH4 production varied among donors, the contrast between low- and- high-CH4 accessions was generally retained, supporting the robustness of accession differences in CH4 production. Several accessions also maintained relatively low CH4 production across contrasting season and water-stress conditions, indicating that favourable CH4 phenotypes were not entirely environment dependent. A multi-trait selection identified ten elite accessions that produced 0.019-0.810 mL CH4/g DM and had 55.2-69.0% IVOMD, whereas ten poor-performing accessions produced 2.80-3.89 mL CH4/g DM and had 50.1-54.7% IVOMD. Differences in hexose fermentation and CH4 produced per unit of fermented substrate further indicated contrasting fermentation efficiency between the groups. These results demonstrate substantial variation in NG and identify accessions with low-CH4 production, favourable digestibility and relatively stable performance across growing conditions. In conclusion, incorporating CH4 yield and digestibility into forage selection could support the development of lower-emission forage cultivars for sustainable climate-smart livestock production.

Background Street food vending is a major source of food supply in Nepal. Poor food hygiene practices among street food vendors pose a health risk by increasing the risk of foodborne diseases. Studies regarding food hygiene knowledge and practices are needed to improve public awareness. Therefore, this study aimed to assess food hygiene knowledge and practice among street food vendors and their associated factors in selected districts of Bagamati Province. Methods A community-based cross-sectional analytical study was carried out among 186 street food vendors of Districts located in central Nepal. Face-to-face interviews were conducted using a semi-structured questionnaire and an observation checklist. Knowledge and practice levels were categorized as good and poor. Chi-square test, Fisher's exact test, odds ratio, and Mann-Whitney U test were applied for data analysis. Results Less than half (39.8%) of the vendors had good food hygiene knowledge, while 91.4% of the respondents reported good food safety and hygiene practices. The education level of street food vendors was significantly associated with food hygiene knowledge (p = 0.001) and food safety and hygiene practices (p = 0.042). Significant association was not seen between overall food hygiene knowledge and overall food safety and hygiene practices levels. Conclusion The study identified a gap between food hygiene knowledge and self-reported food hygiene practices among street food vendors. Education, infrastructure, and regulations among street food vendors play a key role in good food hygiene practices more than knowledge alone. Policymakers should focus on training of food vendors, regular supervision, and monitoring of vending sites to enhance food safety.

Background: Microbiome research has linked microbial community structure to host phenotype, but translating microbiome-based discovery into defined interventions with reproducible long-term host effects remains challenging. We evaluated whether a defined four-member consortium of rumen native microorganisms identified through associations with host phenotype could influence herd retention. Cull outcomes (sold or died vs. retained in herd) were analyzed across 20 controlled commercial dairy trials encompassing more than 40,000 cattle housed across 110 experimental pens, with each trial conducted for at least 5 months. Treatment pens received a defined daily dose of Galaxis Frontier (GF, Native Microbials, San Diego, CA), containing Pichia kudriavzevii, Clostridium beijerinckii, Ruminococcus bovis, and Butyrivibrio fibrisolvens, incorporated into the ration; control and treatment groups were otherwise managed equivalently. Cow-level time to culling was modeled within each trial, accounting for pen-level clustering, and trial estimates were combined using random-effects meta-analysis. Results: Daily treatment with rumen native microbes was associated with a 9% lower hazard of culling across 20 trials (hazard ratio = 0.91, 95% confidence interval: 0.86-0.96; P < 0.001). Between trial heterogeneity was negligible, and treatment effects were not significantly moderated by breed or geographic region. Detailed health records from one trial were examined to further characterize health outcomes associated with GF; treatment was associated with fewer transition-related disorders, particularly ketosis and metritis, whereas mastitis and lameness were largely unaffected. These findings suggest that the health effects associated with GF may be more pronounced for disorders linked to physiological and metabolic adaptation during the transition period than for health events more broadly. Conclusion: Across diverse commercial dairy environments, treatment with a defined consortium of rumen native microbes was associated with improved dairy cow retention. The consistency of this association across independent trials provides large scale evidence that targeted application of native microorganisms can translate into reproducible, long-term outcomes for the host. Complementary health event data suggest that improved transition period health and metabolic stability may contribute to this effect. These findings support the potential for microbiome targeted interventions to influence productive longevity under commercial conditions.

Background: The yak (Bos grunniens) is an essential livestock species in high-altitude alpine ecosystems, serving key roles in meat, milk, and wool production as well as ecological maintenance. However, traditional yak breeding systems face several limitations, including fragmented herd data management, inefficient phenotypic collection, missing pedigree records, and high technical barriers to applying genomic technologies. These factors constrain the widespread application of modern genetic evaluation methods in yak breeding programs. Description: We developed the Yak Genomic Evaluation Platform (YGEP, https://yakbreeding.swun.edu.cn/en/), an integrated digital system for yak genetic selection and herd management. YGEP integrates three core analytical modules: pedigree reconstruction, genotype imputation and genomic breeding value estimation. Benchmarking on 706 yaks showed that GBLUP achieved a prediction accuracy of r = 0.96 for cannon bone circumference (CCB), and KBeagle reached 84.0% imputation accuracy with the combined reference panel. Conclusion: YGEP bridges the gap between advanced genomic analytics and practical yak breeding. By integrating pedigree reconstruction, genotype imputation, and GEBV estimation into a user-friendly cloud platform, YGEP effectively eliminates technical and data bottlenecks. Overall, it provides an accessible, scalable, and standardized solution to advance digital herd management and precision genetic improvement in alpine livestock.

In this work, we investigate the homogeneity of performance in binary classification between the Rotterdam phenotypes and phenotype prediction. For this purpose, a multi-task neural network architecture is designed to improve the prediction accuracy of phenotypes while maintaining the accuracy of binary classification. Using a clinical PCOS dataset which contains unlabeled samples of patients that cannot be attributed to any of the known phenotypes, three models are evaluated: optimized binary Random Forest (RF), six-class weighted class RF, and multi-task neural network (with a common hidden layer and two heads for binary and six-class classifications) optimized by a genetic algorithm. Techniques used for evaluation include ten-fold out-of-fold predictions, paired McNemar tests, bootstrap tests, ablation study, and clustering (Adjusted Rand Index, ARI). Binary RF model reached 90.2% accuracy; however, recall of different phenotypes was significantly unbalanced - from 75.0% of phenotype B to 97.8% of phenotype A. This difference of 22.8 % points was observed (23.3 points) even after excluding phenotype-specific features. While direct phenotype prediction improved recall for common phenotypes, it decreased recall for phenotype B to 43.8% and unclassified cases to 31.5%. Our multi-task model preserved 90.2% binary accuracy and significantly recovered phenotype B recall to 87.5% and unclassified cases to 79.6% (p<0.05). The cluster analysis confirmed the weak correlation between the results and labels (ARI=0.045). The ablation analysis demonstrates that feature selection and class weighting drive this improvement, isolating the specific components responsible for performance gains. We establish that aggregate binary accuracy does not reflect the heterogeneous detection of PCOS phenotypes. Direct phenotype prediction alone redistributes the classification errors, but the multi-task architecture successfully improves the detection of rare phenotypes without compromising binary accuracy. Phenotype-disaggregated evaluations, supported by significance testing and ablation, are essential for clinical classifier research.

Background Micronutrient deficiencies are a major public health problem for women of reproductive age (WRA), especially in conflict settings, such as Yemen. However, there is little information on knowledge, attitudes and practices (KAP) toward micronutrient-rich foods among WRA in Aden Governorate. This study aimed to assess knowledge, attitude and practices of WRA toward micronutrient rich foods attending health facilities in Aden Governorate, Yemen. Methods A facility based descriptive cross-sectional study was conducted among 348 WRA aged 15-49 years who visited selected primary health care and maternal and child health facilities in the eight districts of Aden governorate, Yemen. Data were collected using structured interviewer administered questionnaire which was adapted from guidelines of Food and Agriculture Organization and World Health Organization. Chi-square test and Pearson correlation were used to assess the association between the KAP indicators and sociodemographic characteristics. Results The mean age of the participants was 26.3 {+/-} 8.8 years and 73.9% had high-grade education. In general, 77.9% identified micronutrients as essential for health and 92.8% correctly identified liver/red meat as micronutrient-rich food. However, only 29.6% identified iodized salt as a source of iodine. Although 95.7% perceived supplementation as important, only 9.8% reported regular supplement use. The most commonly reported barrier to access to nutritious food was high food cost (43.4%) and 70.6% perceived healthy foods as too expensive. Restriction in daily consumption was observed for eggs (36.2%), meat/poultry/fish (32.2%) and fruits (13.2%). Education was moderately positively correlated with micronutrient knowledge (r = 0.514, p < 0.001) and age was weakly positively correlated (r = 0.162, p = 0.002). Conclusion WRA in Aden had good knowledge and positive perceptions about micronutrients and supplementation but major gaps in dietary practices and access to affordable nutritious foods. Integrated, culturally appropriate interventions that include nutrition education and improved access to affordable micronutrient-rich foods and appropriate supplementation are needed.

Progesterone governs the establishment and maintenance of pregnancy in ruminants, acting through the nuclear progesterone receptor (PGR) and, potentially, through membrane progesterone receptors (mPRs) of the PAQR family. The cellular distribution of mPRs in ruminant reproductive tissues, however, remains undefined. We generated and validated rabbit polyclonal antibodies against the intracellular C-terminal peptides of bovine PAQR6 (mPR{delta}) and PAQR8 (mPR{beta}), and used them, with quantitative PCR of the uterus and conceptus, to map these receptors in the ovine uterus and conceptus and in the bovine ovary. Both antisera showed high ELISA titer against their immunizing peptides, negligible preimmune reactivity, and complete loss of tissue signal after peptide pre-absorption. By qPCR, PAQR6, PAQR8 and PGR transcript levels did not change significantly across the peri-implantation period in either the conceptus or the endometrium; in contrast, the progesterone-responsive gene ZBTB16 increased markedly in the conceptus (up to ~45-fold by day 21, p < 0.05 at days 17, 19 and 21), whereas FKBP5 showed a numerical increase without reaching statistical significance. Both genes remained relatively stable in the endometrium, indicating conceptus-specific changes in progesterone-responsive gene expression. At the protein level, PAQR6 and PAQR8 localized to the luminal and glandular epithelium of the endometrium across the cycle and early pregnancy, and to follicular (granulosa) and luteal cells of the ovary. Dual immunofluorescence in the ovine endometrium showed that both PAQR6 and PAQR8 are enriched at the apical surface of the luminal epithelium and are spatially distinct from the basolateral adherens-junction marker E-cadherin. These results provide a protein-level map of PAQR6 and PAQR8 in the ruminant reproductive tract, identify the apical epithelial surface as a site of receptor enrichment, and show that the endometrial epithelium retains membrane progesterone receptors while progesterone-responsive gene expression changes dynamically in the conceptus during the peri-implantation period. The validated ruminant-reactive antibodies provide a resource for future functional studies.

Background: There is a growing availability of plant-based alternatives for bovine milk. However, plant-based drinks differ e.g. in protein quantity, protein quality and carbohydrate composition. Knowledge on the impact this has on digestion-related outcomes such as postprandial amino acid (AA) availability and gastric behavior is still limited. Additionally, data on milk alternatives with higher overall protein concentration are lacking. Objective: We aimed to compare the differences in postprandial AA profiles, glycemic response, and gastric behavior between bovine milk and plant-based drinks with or without added protein using representative commercial drinks. Methods: 12 healthy males participated in a randomized crossover study with three treatments. After an overnight fast, participants drank 750 ml of semi-skimmed milk (Milk), a standard oat-based drink (OBD) or an oat-based drink enriched with pea protein (OBD+Pea). Blood samples were taken for measurement of plasma AA, glucose, insulin and FGF21 over 5 hours. Gastric content changes were assessed using magnetic resonance imaging scans for Milk and OBD+Pea over 2 hours. Results: Compared with Milk, both oat-based drinks elicited lower 5-h total AA iAUC (OBD -169706; OBD+pea -139552 microM*min; both P<0.001) and EAA iAUC (OBD -91748; OBD+pea -78199; both P<0.001), with no differences between oat-based drinks. Gastric content volume was lower for OBD+Pea versus Milk from 60-120 min (range -48 - -84 ml, all P<0.001). Compared to Milk, OBD and OBD+Pea increased glucose more from 15-45 min (range 0.86 - 1.73 mmol/l, all P<0.001) and insulin from 30-60 min (1.84 - 2.14-fold, all P<0.001). Conclusions: Protein enrichment in plant-based milk alternatives can be a valuable strategy to increase protein content and improve amino acid composition. However, such products still show faster gastric emptying, lower postprandial essential amino acid and higher glycemic and insulinemic responses compared to bovine milk because of their different carbohydrate and protein content and quality.

Histomonosis, caused by Histomonas meleagridis, is a severe disease in poultry with main pathological lesions in ceca and liver, leading to significant animal suffering and economic losses, with no licensed treatments available. While cecal pathology and microbiota disruption have been documented, the metabolic consequences of infection and vaccination remain poorly understood. Here, we used untargeted LC-MS metabolomics to profile cecal tissue and luminal digesta in a controlled vaccination-challenge chicken trial (n=100, 25 birds/group), including uninfected and unvaccinated but challenged controls, together with birds vaccinated either cloacally, or orally via an edible gel following challenge, across five post-challenge timepoints. Results showed that H. meleagridis infection caused extensive, time-dependent disruption of the cecal metabolome, especially at 7-14 days post-infection (DPI). At DPI 14, 325 tissue and 359 digesta metabolites were significantly altered in unvaccinated challenged birds. Infection was associated with depletion of microbiota-related metabolites, including dicarboxylic acids, tryptophan indoles, phytochemical derivatives, bilirubin catabolites, phenolics, vitamins, and secondary bile acids, alongside accumulation of microbial substrates and fermentation products. Changes in the host-associated metabolites indicated mucosal inflammation, epithelial injury, mitochondrial and amino acid metabolic disturbance, oxidative stress, and membrane lipid remodeling. Cloacal vaccination provided greater metabolic protection than oral vaccination, with 78.3% of disrupted metabolites preserved at DPI 14, compared with 60.7% vaccination by oral gel delivery, pointing towards the importance of vaccine take. By DPI 21, cloacally vaccinated birds showed nearly complete metabolic recovery across most pathway groups, whereas gel-vaccinated birds retained broader metabolic disturbance. These findings reveal histomonosis as a disruption of the cecal host-microbiota metabolic interface and provide a metabolite-level framework for future studies of pathogenesis, vaccine development, and prevention strategies.

Epithelial cells of the female reproductive tract (FRT) form a protective barrier and serve as the first line of defense against sexually transmitted infections (STIs). They recognize pathogens, release cytokines and initiate innate immune responses. Sex hormones 17{beta}-estradiol (E2) and progesterone (P4) fluctuate throughout the menstrual cycle and influence epithelial cell functions. Generally, E2 confers protection against viral STIs like HIV-1 and HSV-2, whereas P4 increases susceptibility. However, the underlying mechanisms remain incompletely understood. MicroRNAs, small non-coding RNAs that regulate gene expression by inhibiting translation of target mRNAs, significantly influence viral infections, by promoting or restricting viral pathogenesis. Here, we examined the effect of sex hormones on microRNA expression in vaginal epithelial cells (VECs), the first responders to pathogens in the FRT. Using next generation sequencing analysis, the effect of physiological concentrations of E2 or P4 on microRNA transcription profile of VECs grown in air-liquid interface cultures was characterized. Thirteen miRNAs were uniquely regulated by E2 and 102 by P4, with 29 miRNAs that overlapped between both. The top four differentially expressed miRNAs from both treatments were validated by RT-qPCR. The mRNA targets of unique E2-miRNAs were involved in adherens junction, tight junction and chemokine signaling pathways; whereas the mRNA targets of unique P4-miRNAs were involved in MAPK signaling pathway. Interestingly, several miRNAs regulated by E2- (eg. hsa-miR-9-3p, hsa-miR-200c- 5p, hsa-miR-548c-5p) and by P4 (eg. hsa-let-7f-5p, hsa-miR-16-5p, hsa-miR-27b-3p) targeted genes related to cell cycle pathway. Together, these findings indicate that microRNAs regulated by E2 and P4 influence mRNAs in the pathways associated with cell adhesion, cell growth and development and innate immune responses, critical for maintenance of epithelial barrier integrity and regulating pathogen susceptibility in the vaginal tract. These hormone-regulated miRNAs may serve as potential targets for therapeutic interventions.

High calorie intake of unhealthy foods are the main factor of overweight and obesity. In food environment, food retailers are the biggest player in shapping consumers purchase pattern. In this South East Asia Obesogenic Food Environment (SEAOFE) study, we aimed to to explore retailer perspectives on stocking decisions and barriers to stocking healthy food products in urban poor areas of Kuala Lumpur, Malaysia. The study was conducted using purposive sampling at three parliaments which have the highest urban poor population. 35 retailers who involved in procurement, retail operations, and product placement, with at least one year of experience, were selected as participants of this study. The interview were conducted face-to-face and been transcribed verbatim for data analysis. Nine key themes emerged regarding factors affecting stocking decisions: 1) consumer demand, 2) consumer satisfaction and feedback, 3) profitability, 4) forecast software, 5) fast-moving products, 6) product healthiness, 7) availability, 8) seasonality, and 9) e-commerce; while four key barriers in stocking healthy food products included 1) price, 2) shelf life, 3) consumer demand and preferences, and 4) availability of supply. This study showed that the urgency of targeted interventions to improve healthy food supply through collaboration from retailers, suppliers and policymakers. By addresing the issue will help to increase consumption of healthy foods and improve the health of the residents within the urban poor areas of Kuala Lumpur.

Thailand is experiencing a rapid rise in childhood obesity, increasing the risk of early-onset non-communicable diseases. Although whole grains support weight management and reduce non-communicable diseases risk, consumption remains low in Thailand and is insufficiently addressed in the Thai Food-Based Dietary Guidelines. Whole grains are also rarely included in the National School Lunch Program. Given its high participation rate, the school lunch program represents a strategic platform for increasing whole-grain intake and fostering lifelong healthy eating habits. This study aimed to identify facilitators and barriers to increasing whole-grain provision in Thai elementary schools from the perspectives of adult stakeholders We conducted in-depth interviews guided by the Integrated Behavioral Model with 19 teachers and contracted school cooks from two Thai elementary schools, and with 10 faculty and staff members at Mahidol University. Interviews explored perceived norms regarding student whole grain consumption, attitudes toward whole grain, and perceived facilitators and barriers to whole grain provision in schools. Data were analyzed using framework analysis, combining the Integrated Behavioral Model-informed thematic categorization with inductive coding. Transcripts were independently coded by two researchers to develop an Integrated Behavioral Model-grounded conceptual framework describing factors influencing intentions to increase whole grain provision in Thai elementary schools. Participants were predominantly female (93.1%) and highly educated (89.7% with a bachelors degree or higher). Most supported incorporating whole grains into school meals, citing their health benefits for schoolchildren. However, participants reported low WG intake among schoolchildren, attributable to limited availability at both schools and homes. Two primary barriers to whole grain provision were identified. The first was student-level resistance, including unfamiliarity and sensory aversions, particularly toward brown rice. The second involved systemic challenges, including the absence of regulatory guidance promoting whole grain inclusion and operational constraints related to contracted food services and inflexible menu systems. Multi-level engagement among schools, households, and government agencies was identified as a key facilitator of increased whole grain provision in school meals.? These findings indicate that although teachers and cooks are receptive to whole-grain provision in school meals, translating this support into practice will require coordinated, multisectoral action to address persistent structural barriers at the student, school, and policy levels. Future research should incorporate childrens perspectives and examine a broader range of school settings to inform the design of effective, sustainable strategies for whole grain integration.

Background: Added sugars during early development may influence adiposity, but whether associations vary by developmental timing and child sex is unclear. Objectives: To determine whether child sex modifies associations of added sugars and fructose exposure from pregnancy through 24 months with body composition at 24 months. Methods: This secondary longitudinal analysis included 130 mother-child dyads. Maternal diet was assessed during mid-pregnancy and child diet at 2wk and 6mo (infancy), and 12mo and 24mo (toddlerhood). Multivariable regression tested sex-by-exposure interactions for anthropometric outcomes; 42 children had usable dual-energy X-ray absorptiometry (DXA) data. Results: Prenatal added sugars were more positively associated with percentage body fat and total and regional fat mass in girls than in boys. Prenatal fructose was more positively associated with waist circumference in boys. During infancy, added sugars were more positively associated with percentage body fat and total and regional fat mass in boys, whereas fructose was more positively associated with fat-free mass in girls. Conclusions: Associations of early-life sugars exposure with body composition varied by developmental timing and child sex. Findings from the smaller DXA subset are exploratory and require replication but suggest that susceptibility to early dietary sugars may differ across developmental windows.

Background: University life can influence food choice even among physically active students. This study examined nutritional awareness, emotional, external, and restrained eating, body mass index (BMI), and estimated resting metabolic rate (RMR) in physical education students, and assessed differences by sex and academic level. Methods: A descriptive, analytical and correlational design was used. The sample included 160 physical education students (95 women and 65 men; mean age 20.42 {+/-} 1.53 years) from An-Najah National University. Nutritional awareness was assessed with a 15-item scale, and eating behaviour with the 33-item Dutch Eating Behavior Questionnaire. BMI was calculated from measured height and body mass, and RMR was estimated using an equation developed for athletes. Correlations, group comparisons, robust interaction models, and hierarchical regression were applied. Results: Nutritional awareness was high (3.73 {+/-} 0.81), whereas emotional (3.11 {+/-} 1.18), external (3.20 {+/-} 0.95), and restrained eating (3.25 {+/-} 1.11) were moderate. Higher awareness was associated with less emotional (r = -0.617) and external eating (r = -0.652), but greater restrained eating (r = 0.644; all q < 0.001). These associations remained after adjustment. Men reported more external eating, while women reported more restrained eating. Eating patterns also varied by academic level. BMI and estimated RMR showed weaker and less consistent associations with eating behaviour. Conclusions: Nutritional awareness was closely associated with reported eating behaviour, whereas BMI and estimated RMR were less informative. Sex and academic-level differences suggest that nutrition education for physical education students may be more useful when adapted to specific behavioural patterns. The findings represent associations and group differences and do not establish causal relationships. Keywords: physical education students; nutritional awareness; eating behaviour; emotional eating; external eating; restrained eating; body mass index; resting metabolic rate

Aim: This study aimed to estimate the prevalence and distribution of minimum dietary diversity for women (MDD-W) and underweight/thinness and examine sociodemographic and dietary factors associated with underweight/thinness among Cambodian women aged 15-49 years. Methods: We conducted a cross-sectional analysis of 9,181 women aged 15-49 years (9,262.5 weighted) from the 2021-22 Cambodia Demographic and Health Survey. Survey-weighted descriptive statistics and provincial mapping were used to describe MDD-W, food-group consumption and underweight/thinness. Rao-Scott chi-square tests assessed group differences, and survey-weighted logistic regression estimated factors associated with underweight/thinness. MDD-W and household wealth were retained a priori in the adjusted model, while other covariates with an overall crude Wald P value <0.20 were included. Results: The analysis included 9,181 women (9,262.5 weighted). Overall, 57.2% achieved MDD-W, and 6.7% had underweight/thinness; 22.8% were overweight, and 5.7% had obesity. Underweight/thinness was highest among women aged 20-24 years (17.7%). Achieving MDD-W was not associated with underweight/thinness in crude analysis (OR 0.93, 95% CI 0.74 to 1.17; P=0.545). In the adjusted analysis, the association remained imprecise (aOR 0.86, 95% CI 0.67 to 1.09; P=0.219). Provincial estimates showed geographic variation in both MDD-W and underweight/thinness. Conclusion: More than half of Cambodian women achieved MDD-W, while underweight/thinness coexisted with overweight and obesity. MDD-W was not clearly associated with underweight/thinness. These findings support monitoring dietary diversity, anthropometric status, and socioeconomic inequalities together in women's nutrition.