Journal of Dairy Science

Naturally occurring variation in the bovine mannose receptor C-type 1 gene (MRC1) may shape macrophage responses to Mycobacterium (M.) bovis, a key driver of bovine tuberculosis (bTB). We identified four coding region SNPs in MRC1 across Bos taurus (Holstein Friesian, Brown Swiss) and Bos indicus (Boran, Sahiwal) cattle breeds, including a non-synonymous variant, rs380943118 (c.2963G>A; Ser988Asn) in C-type lectin-like domain (CTLD) 6, most prevalent in Sahiwal cattle. Structural modelling suggested that the S988N substitution, which is spatially separated from the monosaccharide binding site of CTLD4, might indirectly affect glycan binding, perhaps through a conformational change in the receptor. Monocyte-derived macrophages upregulated MR expression during differentiation, with heterozygous (G/A) animals showing higher MR expression and increased uptake of GFP-M. bovis BCG, although differences were not statistically significant. Anti-CD206 blockade did not inhibit BCG internalization, either indicating that this specific antibody did not bind to a CTLD involved in ligand binding or that MR is not the sole entry receptor. These results highlight naturally occurring MRC1 polymorphisms that may influence MR structure and macrophage function, providing a foundation for future studies to assess their role in bTB susceptibility.

Metabolites from Plantago lanceolata (plantain) biomass have been linked with biological nitrification inhibition (BNI) in soil. After grazing, leaf metabolite chemistry is altered via digestion, and a suite of secondary metabolites are then delivered onto soil via dung and urine. The purpose of this study was to establish if urine from sheep grazed on plantain had BNI activity when added to pasture soil, and to identify the metabolite profile(s) that most likely contribute to the BNI effects observed. Groups of sheep (n=5) were grazed on one of nine different plantain cultivars in autumn and spring with analysis of leaf material, urine, soil incubation and BNI bioassay data used to identify potential metabolite candidates implicated with BNI. The urinary nitrogen and metabolite composition of sheep fed plantain varied significantly between cultivars and season. After 28 days of incubation, all soil microcosms treated with plantain-derived urine had up to 35% less nitrate than comparative ryegrass urine controls in both seasons, except one in autumn. The key phytochemistry associated with lower soil nitrate concentrations was phenylethanoid and iridoid glycosides resulting in a higher output of glucuronidated, methylated and sulfated secondary metabolites in the urine. Among 19 secondary metabolites identified in the urine, hydroxytyrosol-related metabolites as well as catechol glucuronide, 2-methoxyphenyl sulfate and guaiacol-{beta}-D-glucuronide appear to be the most likely target compounds with respect to the BNI effects observed. Variation in metabolites from different plantain cultivars affected the ratio of metabolite derivatives in urine, which ultimately affected soil nitrification rates. Cultivar phytochemistry is therefore an important consideration with respect to BNI under urine patches. HighlightsO_LISheep grazing different plantain cultivars had different urine compositions C_LIO_LIUrines elicited biological nitrification inhibition (BNI) in soil and in vitro C_LIO_LIDifferent BNI response was related to differential expression of urine metabolites C_LIO_LIKey urine metabolites associated with BNI are derived from glycosidic compounds C_LI

Anthelmintic resistance in cyathostomins is escalating worldwide, threatening equine health and highlighting the need for sustainable, ecology based parasite control strategies. Chicory (Cichorium intybus, Puna II) has emerged as a promising antiparasitic forage, yet its broader effects on the equine holobiont, parasites, microbiota, and host physiology remain poorly understood. We conducted a 32 day longitudinal grazing trial in young horses to assess how chicory affects parasitological outcomes, gut microbial ecology, nemabiome composition, behaviour, and host physiological and immune responses. Twenty-six naturally infected Anglo-Arabian horses were monitored weekly, with 13 grazing a chicory-based sward and 13 grazing a permanent pasture. Clinical parameters, body weight, and serum biochemistry remained stable across treatments, indicating that chicory was well tolerated. Immune profiles showed limited variation, although IL 10 increased in chicory fed horses, suggesting subtle immune modulation. Behavioural observations revealed no signs of discomfort and indicated slightly enhanced social interactions in the chicory group. Chicory grazing produced a marked reduction in cyathostomin egg excretion, accompanied by species specific shifts in nemabiome composition. Several cyathostomin taxa, including Cylicocyclus ashworthi, C. leptostomus, and C. nassatus, declined in chicory fed horses, whereas certain Cylicostephanus spp increased, indicating differential sensitivity rather than uniform suppression. Concomitantly, chicory induced profound ecological changes in the gut microbiota, including reduced alpha diversity, increased beta dispersion, and destabilised individual microbial trajectories. Several bacterial lineages, particularly Oscillospiraceae, Clostridiaceae, Lachnospiraceae, and Bacteroidales, were differentially enriched, reflecting a functional reorganisation of the intestinal ecosystem. Together, these findings demonstrate that chicory reduces parasite fitness, reshapes nemabiome composition, and alters gut microbial ecology while maintaining host physiological stability. Chicory thus emerges as a promising ecological tool for parasite control, capable of modulating the equine holobiont in ways that complement and potentially reduce reliance on conventional anthelmintic strategies. However, because its effects on gut microbial ecology remain uncertain, and may include shifts resembling dysbiosis, future studies are needed to monitor microbial dynamics more closely and clarify the long term ecological consequences of chicory grazing.

The Brown Swiss (BS) cattle breed is one of the major Swiss dairy breeds. Intensive selection and the widespread use of few elite sires in artificial insemination have increased inbreeding and the occurrence of deleterious recessive alleles in the homozygous state. Analyzing life trajectories in large, genotyped cohorts can identify hidden recessive disorders that are difficult to detect using traditional case-control association testing. Long-read DNA sequencing enables precise detection of causal alleles, including structural variants. This study aimed to (1) identify cryptic recessive loci affecting rearing performance in Swiss BS cattle, (2) evaluate their impact on survival, (3) characterize the associated phenotype, (4) identify the causal variant using long-read whole-genome sequencing, and (5) assess its functional impact. Using Homozygous Haplotype Enrichment/Depletion (HHED) mapping, we identified a risk haplotype (BH39) on chromosome 15 spanning from 16,276,819 bp to 16,446,984 bp that was associated with increased juvenile mortality within the first 180 days of life when present in the homozygous state. The BH39 occurred at a frequency of approximately 4.5% in Swiss BS cattle and 5.3% in German and Austrian BS cattle, and homozygous carriers exhibited a significantly reduced first-year survival rate. Five females homozygous for BH39 underwent clinical examination. They all showed recurrent respiratory disease, impaired growth, poor body condition, rough hair coat, and brown-discolored teeth. Pathological examination revealed bronchopneumonia and eosinophilic enteritis. Clinicopathological findings indicated failure to thrive and immunodeficiency. Long-read WGS of two BH39 homozygous calves revealed a private homozygous coding variant that was in high linkage disequilibrium with BH39. The identified structural variant was an insertion of a large transposable element (10.4 kb ERVK[2-1-LTR]) into the third exon of ALKBH8 (NM_001080341.2 c.267_268indel). Full-length RNA sequencing of cerebellum and liver from a homozygous calf revealed that the endogenous retrovirus (ERV) insertion introduces a cryptic transcription termination signal, truncating ALKBH8 mRNA. This study demonstrates that exploring population-scale genomic data and mining thousands of life-history records, followed by veterinary follow-up evaluations and molecular genetic analyses, provides an effective strategy for identifying cryptic recessive disorders that shorten the lifespan of cattle. The findings provide strong evidence that the ERV insertion into the coding sequence of ALKBH8 represents a loss-of-function variant that causes a previously undescribed recessive disorder that results in increased rearing loss. Interpretive summaryWe identified a recessive disorder in Brown Swiss cattle that causes retarded growth, recurrent infections, immunodeficiency, and increased mortality during the first year of life. Using population-scale genomic data, clinical investigations, and long-read sequencing, we linked the disorder to an exonic transposable element insertion disrupting ALKBH8. The identification of the causal variant now enables direct genetic testing and the implementation of genome-based mating strategies to avoid carrier-by-carrier matings and, consequently, prevent the birth of affected homozygous offspring. We demonstrate the utility of integrating large-scale breeding records, veterinary phenotyping, and advanced genomics to identify hidden defects affecting livestock health and productivity.

Background: Iron deficiency (ID) affects up to 40% of pregnant women in the third trimester, even in highly resourced and iron-supplemented populations, with adverse consequences for maternal health and long-term offspring development. Psychological stress may compromise iron status through hypothalamic-pituitary-adrenocortical (HPA) axis dysregulation and inflammation, but no study has directly examined cortisol in relation to iron status across human pregnancy. Objective: This longitudinal study examined associations between HPA function and maternal iron status across pregnancy and tested whether IL-6 and CRP mediated the relationship between cortisol and ferritin across gestation. Methods: One hundred sixty-eight pregnant Black women with Medicaid insurance completed up to four laboratory assessments across pregnancy. Salivary cortisol was measured before and in response to the Trier Social Stress Test, yielding basal and reactive cortisol indices. Serum ferritin, IL-6, and CRP were collected at each visit. Trimester-specific regression models examined cortisol reactivity in relation to ferritin; linear mixed-effects models with moderated mediation tested whether basal cortisol predicted ferritin via inflammation. Results: Higher cortisol reactivity was associated with lower ferritin specifically in the third trimester (std. {beta} = -0.197, p = .004). Higher basal cortisol predicted a steeper IL-6 rise across gestation (p = .002), and IL-6 was positively associated with ferritin (b = 0.236, p = .006), consistent with inflammatory iron sequestration. The indirect effect of basal cortisol on ferritin via IL-6 was statistically significant, and higher basal cortisol was negatively associated with cortisol reactivity in the third trimester. No pathway was observed through CRP. Conclusion: Greater cortisol reactivity predicted lower third-trimester ferritin, a pattern that suggests cumulative iron depletion, atypically sustained HPA reactivity in late pregnancy, or both. To our knowledge, this is the first prospective study linking cortisol reactivity to iron status across human pregnancy, identifying maternal stress physiology as a novel target for understanding and addressing gestational iron deficiency.

Background The placenta serves a critical role in nutrient uptake and waste elimination for the developing fetus. The placenta is also responsible for the uptake and/or exchange of xenobiotics, including medications, between the maternal and fetal bloodstreams. An estimated 40-80% of women take medications or drugs during pregnancy for a variety of conditions. Very little is understood about fetal drug and nutrient exposure during pregnancy and how it may change over the course of fetal development. Objective This study aimed to characterize the abundance of transport proteins in placental tissue, which are important in modulating fetal nutrient and drug exposure, over the duration of pregnancy. Mass spectrometry-based global proteomic analysis revealed trends in the expression of thousands of proteins throughout gestation. Focusing on the membrane-associated proteome enabled an increased emphasis on the solute carrier and ATP-binding cassette families of transporter proteins that are critical for nutrient and xenobiotic transport across the maternal-fetal barrier. Study Design Using data-independent acquisition proteomics, relative abundance of proteins in placental tissue samples was profiled across all three trimesters of pregnancy (Trimester 1 = 16, Trimester 2 = 9, and Term = 9). Membrane fractions were generated to enrich membrane-associated proteins for proteomic analysis. Placental samples were grouped into randomized batches for membrane fraction generation and mass spectrometry analysis. Proteomic search results from each batch were imported into the R programming environment from Skyline, concatenated, and normalized as one data set for downstream analysis. Results A total of 6,331 proteins were detected across all samples with 4,210 proteins identified in every sample. Pathway analysis revealed that as gestational age increases, membrane-associated proteins involved in more complex metabolic pathways increase in relative abundance while those involved in extracellular remodeling events and simple organic ion transport tended to decrease. A total of 139 solute carrier and ATP-binding cassette transport proteins were identified in all samples, and 80 were identified in every sample. In general, membrane-associated proteins, including solute carrier and ATP-binding cassette transport proteins, were significantly enriched in placental tissue collected during early gestation compared to term placental tissue. Conclusion This study presents a comprehensive profiling of membrane-associated proteomic changes during gestation and identifies significant gestational age associated abundance changes at the protein level in several transport protein families. The application of data-independent acquisition global proteomic techniques enabled in-depth analysis of thousands of proteomic changes across pregnancy in a single experiment. These data provide critical information to support future studies into the understanding of fetal exposure to xenobiotics and nutrients circulating in the maternal bloodstream.

Dietary protein restriction challenges the allocation of amino acids to growth and other physiological functions and therefore requires coordinated metabolic adaptation. Domestic pigs provide an informative system in which to study such responses, because nitrogen retention directly affects lean growth and can be quantified accurately under controlled feeding and housing conditions. Under reduced-protein diets, pigs differ in how effectively they retain nitrogen, and this variation has a genetic basis, making them well suited to investigate the molecular regulation of nitrogen use efficiency (NUE). Here, we characterise differential gene expression and enriched pathways in liver and skeletal muscle of more than 80 pigs with two divergent NUE phenotypes (high and low) maintained under the same protein-reduced, ad libitum dietary conditions. The two NUE phenotypes were clearly distinct at the transcriptomic level, with 177 differentially expressed genes in the liver and 133 in the muscle. In the liver, differential expression and enrichment analyses indicate reduced amino acid catabolism, lower inflammatory and detoxification activity, and a metabolic state that favours lipid processing and insulin-related regulation over the use of amino acids as energy sources. In skeletal muscle, they point to reduced lipid uptake, lower reliance on amino acid oxidation, and a greater emphasis on protein synthesis, translational regulation, mitochondrial energy metabolism, and growth-related processes. These gene-level patterns were supported and extended by pathway and gene-set enrichment analyses. Together, the results suggest that high and low-NUE pigs differ through coordinated, tissue-specific molecular adaptations. Overall, variation in NUE appears to reflect coordinated, tissue-specific differences in how nutrients are allocated between energy use, storage, and lean tissue growth.

Background: Yeast (Saccharomyces cerevisiae) is a model organism in agricultural and industrial fermentation with nutritional benefits, yet less is understood about its nutritional value for supporting whole-body protein synthesis in vivo, and its comparison to animal-based protein. Objective: This study aimed to determine the effect of microbial-based protein (yeast) compared to a high (whey) and low (collagen) quality animal-based protein on the ability to support whole-body protein synthesis using the indicator amino acid oxidation technique. Methods: Thirteen healthy participants (M: n=6, 24{+/-}4 yr; F: n=7, 27{+/-}7 yr) consumed eight hourly yeast (Y), whey (W), or collagen (CH) protein beverages at 0.9 g{middle dot}kg-1{middle dot}d-1 supplemented with L-[1-13C]phenylalanine in a randomized, cross-over, counterbalanced design. Breath and urine were collected to measure fraction of expired 13CO2 (F13CO2) and [1-13C]Phe oxidation (PheOx) as inverse correlates of whole-body protein synthesis. An a priori 20% noninferiority margin was used to determine equivalency between protein sources. A Visual Analog Scale (VAS) was provided to assess fullness and hunger by protein sources. Results: Protein source had no effect on F13CO2 (P=0.15) or PheRa (P=0.10). PheOx was greater in CH compared to both Y (14.94{+/-}2.16 vs. 12.93{+/-}2.80 mol{middle dot}kg BM-1{middle dot}h-1, P<0.05) and W (14.94{+/-}2.16 vs. 12.50{+/-}2.75 mol{middle dot}kg BM-1{middle dot}h-1, P<0.05) whereas there was no significant difference between Y and W (P=0.59). Change in PheOx (mean difference [90% CI]) between CH and W (2.44 mol{middle dot}kg BM-1{middle dot}h-1 [0.98, 3.90], P<0.05) and between Y and W (0.44 mol{middle dot}kg BM-1{middle dot}h-1 [-1.34, 2.21], P>0.99) confirmed respective inferiority and noninferiority to W compared to the 20% noninferiority margin (2.50 mol{middle dot}kg BM-1{middle dot}h-1), while Y was superior to CH (-2.00 mol{middle dot}kg BM-1{middle dot}h-1 [-3.01, -0.99], P<0.05). Protein source did not significantly influence satiety (P=0.32) or hunger scores (P=0.15). Conclusion: EAA-enriched whey and yeast protein similarly reduced PheOx as compared to EAA-deficient collagen hydrolysate, suggesting these proteins were similarly superior in supporting whole-body protein synthesis.

Bovine embryo in vitro production (IVP) is characterised by low efficiency and variable outcomes. Monitoring early embryonic development by videomicroscopy revealed substantial morphokinetic heterogeneity in the first four embryonic cycles (EC, conventionally referred to as the 2-, 4-, 8- and 16-cell stages). Morphokinetic analysis offers a promising approach to characterize divergent developmental trajectories and provide a better understanding of underlying molecular mechanisms. We developed a Random Forest classification system (Bovine Embryo Analyser based on Morphokinetics: BEAM) to predict embryo phenotype. It is based on morphokinetic variables collected from the 1st to the 4th EC and predicts four blastocyst categories (EHB: Early Hatching Blastocyst, HB: Hatching Blastocyst, SSB: Subtle Developmental Shift Blastocyst, ADB: Arrhythmic Development Blastocyst). Classification performance on an independent dataset was good (F1 score = 0.59; Accuracy = 0.78), indicating that the BEAM can be useful for embryo development studies. The BEAM was further applied to embryos submitted to 4.3 days of culture and having completed the 4th EC (16-32 cells). RNA sequencing was performed on sixteen samples (4 x 8 pooled embryos/category). The ADB category was significantly enriched in transcripts involved in the regulation of transcriptional activity compared to the EHB category. In addition, in the ADB category, 22.7% (n = 185/816) of the upregulated genes were of maternal origin while only 6.2% were of embryonic origin (n = 54/816). In conclusion, despite being at a comparable developmental stage and transcriptionally competent, the ADB category showed delayed maternal transcript degradation suggesting delayed transition to embryonic transcriptional autonomy. Illustrated Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=138 SRC="FIGDIR/small/733532v1_ufig1.gif" ALT="Figure 1"> View larger version (45K): org.highwire.dtl.DTLVardef@13a0958org.highwire.dtl.DTLVardef@13b88a1org.highwire.dtl.DTLVardef@18707c9org.highwire.dtl.DTLVardef@105759_HPS_FORMAT_FIGEXP M_FIG C_FIG Summary SentenceIn vitro produced bovine embryo morphokinetic patterns allow prediction of four blastocyst categories (up to the 4th embryonic cycle) and are associated with distinct transcriptomic profiles at embryonic genome activation in competent embryos.

Introduction: Low and middle-income countries suffer from a high burden of undernutrition. Fetal malnutrition negatively impacts a newborn's body composition, metabolism, and enzymatic processes, predisposing the newborn to malnutrition during childhood period. This study aims to assess the magnitude of fetal malnutrition and its predictors in southern Ethiopia. Methods: A facility-based cross-sectional study was conducted among 423 pairs of mother and term newborn delivered from February 1- 28, 2025, at five randomly selected public hospitals in southern Ethiopia. The Clinical Assessment of Nutrition (CAN) Score is used to assess fetal malnutrition. Logistic regression was employed to identify predictors of fetal malnutrition. Statistical significance of the association was declared at p < 0.05. Results: Among 423 newborns included in the study, 60 (14.1%) had fetal malnutrition. Newborns born to women with a placental weight of 519 grams or less had ten times higher odds of fetal malnutrition compared to their counterparts (AOR=9.795, 95% CI: 4.881-19.657). Dietary counselling during pregnancy reduced odds of fetal malnutrition by 62.3% (AOR=0.377, 95% CI: 0.162-0.877); similarly, an extra meal during pregnancy was associated with reduced odds of fetal malnutrition by 71.6% (AOR=0.284, 95% CI: 0.131-0.616). Newborns delivered from women who had a MUAC (Mid Upper Arm Circumference) >22 cm had 75.7% lower odds of fetal malnutrition (AOR=0.243, 95% CI 0.074 -0.797), whereas maternal chronic medical illness increased the odds by threefold (AOR=3.419, 95% CI: 1.269-9.153). Conclusion: There is a high magnitude of fetal malnutrition in the study area. Placental weight, dietary counselling, extra meals during pregnancy, MUAC and chronic medical illness were predictors of fetal malnutrition, signifying the need for a comprehensive approach targeting maternal nutrition during pregnancy

Background and aims: Maternal overweight and obesity are increasing worldwide, including Nepal. This study assessed BMI trajectories from early pregnancy to one year postpartum and trends in overweight and obesity over the past two decades in Bhaktapur, Nepal. Methods: In the most recent study, BMI was measured in 800 Nepalese women at three time points: at early pregnancy, 6 and 12 months postpartum (2017-2021). The prevalence of undernutrition, overweight, and obesity was estimated using the World Health Organization and the Asian specific cut-offs. Long-term trends were assessed by comparing these findings with three population-based studies conducted in Bhaktapur between 2001 and 2021 among 2400 women at similar life stages. Results: Mean (SD) BMI increased from 23.7 (3.0) kg/m^2 in early pregnancy to 26.1 (3.3) kg/meter squre at 6 and 25.2 (3.3) kg/m^2 and 12 months. The prevalence of overweight increased from 32.9% in early pregnancy to 48% at 6 months. Using the Asia-specific cut-offs, the prevalences were higher. Results from the three previous population-based studies demonstrated an upward trend where postpartum overweight increased from 11.4% in 2001- 2002 to 44.6% in 2017- 2021. The obesity prevalence rose from 1.8% to 10.9% during this period. Conclusion: Overweight and obesity among Nepalese women have risen dramatically over the past two decades, with postpartum overweight increasing nearly fourfold and obesity more than sixfold. These findings highlight the need for interventions to prevent excessive weight retention and reduce adverse health outcomes.

Consumer-grade bioelectrical impedance analyzers have become widely used for body composition assessment, yet their accuracy varies considerably across devices. The Hume Pod is a popular consumer-grade analyzer marketed as being highly accurate, but independent validation is lacking. The purpose of this study was to evaluate the reliability and validity of the Hume Pod relative to both a four-compartment (4C) model and dual-energy X-ray absorptiometry (DXA). Sixty-seven adults (42 females, 25 males; age 37.2 +/- 13.5 years, body mass index: 24.6 +/- 4.9 kg/m2, body fat percentage [BF%]: 26.4 +/- 10.2%) completed duplicate Hume Pod assessments alongside DXA and 4C evaluations. Reliability was evaluated using the technical error of measurement (TEM) and intraclass correlation coefficients (ICC). Validity was assessed using equivalence testing, Lin's concordance correlation coefficient (CCC), standard error of the estimate (SEE), Bland-Altman analysis, and additional tests. The Hume Pod demonstrated strong reliability, with ICCs >/= 0.993 and TEMs of 0.8% for BF% and 0.6 kg for fat mass (FM) and fat-free mass (FFM). Relative to the 4C model, BF%, FM, and FFM estimates were statistically equivalent (all p<0.05), with strong agreement (CCC=0.95-0.98), low SEE values (3.1%, 2.3 kg, and 2.2 kg, respectively), moderate limits of agreement (+/-6.1%, +/-4.5 kg, and +/-4.5 kg), and no proportional bias. Compared with DXA, generally strong agreement was also observed. These findings indicate that the Hume Pod demonstrates strong reliability and validity compared with laboratory reference methods for body composition estimation, supporting its potential use as a consumer body composition assessment.

Objectives: Research on mental health in agriculture has increased in recent years; however, it remains largely focused on farmers themselves and is predominantly male-oriented. The mental health of farm wives and partners, many of whom play integral roles in farm operations, business management, and family life, remains difficult to characterise. This study therefore aims to explore the prevalence and causes of mental health challenges among farm wives and partners, and to investigate their use of, and barriers to, mental health support services. Methods: Quantitative data was collected using over 450 structured questionnaire responses that assessed mental health prevalence, contributing stressors and support service utilisation. Results: Findings indicate that there is a high prevalence of mental ill health amongst farm wives, seemingly due to industry stressors and support role overwhelm. Interpersonal relationships played a significant role in the types of mental distress experienced and highlighted the toll that farm life can take on farm wives' social and emotional connections. Despite a range of formal and informal support services being available, and effective when used, significant barriers to accessing these services were identified, including both practical difficulties and self-stigmatisation due to cultural beliefs. Conclusions: Farm wives and partners experience substantial mental health burdens linked to their diverse and often underrecognized roles within agricultural systems. In future, targeted interventions are needed to reduce stigma, improve service accessibility, and recognize women's contributions within farm enterprises. Further research and dedicated investment are also essential to better understand and help improve the mental health of this overlooked population within agricultural industries.

The respiratory microbiome plays important roles in poultry health, immune regulation, and pathogen resistance, yet its development throughout the broiler production cycle remains poorly understood. This study investigated temporal changes in the tracheal microbiome of broiler chickens across production phases. Tracheal samples were collected during the starter (day 12), grower (day 21), early finisher (day 26), and late finisher (day 35) phases and analyzed using 16S ribosomal RNA gene sequencing. Tracheal microbial richness, diversity, community structure, and taxonomic composition changed significantly across broiler production stages, including starter, grower, early finisher, and late finisher feeding phases. Alpha diversity increased progressively throughout production, with significant increases in richness, diversity, and phylogenetic diversity during later stages. Beta diversity analysis revealed distinct microbial communities associated with each production phase, with starter-phase samples clearly separated from later phases. Taxonomic profiling showed dominance of Proteobacteria during the starter and grower phases, with enrichment of Methylobacterium-Methylorubrum and Pseudomonas during the starter phase and of Escherichia-Shigella during the grower phase. In contrast, the finisher phases exhibited reduced Proteobacteria abundance and increased Firmicutes and Actinobacteriota, including Lactobacillus, Ligilactobacillus, Faecalibacterium, Streptococcus, Staphylococcus, Romboutsia, and Corynebacterium. Overall, the tracheal microbiome underwent progressive maturation, shifting from a Proteobacteria-dominated community to a more diverse, complex, Firmicutes-rich ecosystem. These findings provide new insights into the development of the respiratory microbiome in broiler chickens and may support strategies to improve poultry respiratory health. Because dietary transitions occurred concurrently with age progression, the observed microbiome shifts should be interpreted as production-stage-associated changes rather than diet-specific effects.

Background: Despite growing evidence that personalised nutrition (PN) can improve dietary behaviour and health outcomes, the implementation of PN food services in real-world settings remains largely unexplored. This study evaluated the feasibility of a comprehensive PN service for delivering personalised nutrition products (PNPs) in a military setting and assessed consumer acceptance following repeated consumption. Methods: A complete PN service was developed to design, produce, and distribute personalised nutrition products (PNPs). Twenty-five participants from the Royal Netherlands Army took part in a feasibility study, using a single-cohort pre-test post-test design. Each participant consumed a PNP on 10 consecutive days during a two-week combat training exercise. Product liking was assessed through daily questionnaires, while broader user experiences were assessed at baseline (t=0), one week follow-up (t=1) and two-week endline (t=2). Results: The PN service was succesfully implemented during a real-life military training exercise. In total, 229 PNPs were produced and distributed. Participants evaluated all aspects of the PN service from just below neutral to slightly positive (3.4 to 5.0 on 7 point scales). Across the different product combinations, mean overall liking and all sensory attributes were slightly positive (5.7 to 6.1 on 9 point scales). Overall liking did not change over time (F (9,194) = 1.242, p=0.27). Attitudes toward a PNP service in the army decreased over time, but remained just-above neutral at study end. Conclusions: This study demonstrates the feasibility of a personalised nutrition service in a military setting. Both the service and PNPs received modestly positive evaluations, while repeated exposure did not change overall liking of the products. Although further optimisation of the service is warranted, these findings support the potential for broader implementation of PN services in military and other high performance environments.

Background Gestational diabetes mellitus (GDM) affects 1-in-7 pregnancies globally and is associated with significant short- and long-term health consequences. Although health behaviour change interventions can effectively reduce these risks, a significant implementation gap exists in translating this evidence into routine practice. Bump2Baby and Me (B2B&Me) was a mobile health (mHealth) coaching intervention provided to women at-risk of GDM from early pregnancy through to 1-year postpartum. B2B&Me Plus (B2B&Me+) aims to refine, implement and evaluate the implementation of this personalised intervention across 4 countries (Ireland, Spain, Poland and Norway) with differing health systems and population profiles. Methods This study employs a hybrid type 3 implementation-effectiveness design using a non-randomised ABA block approach within a longitudinal cohort. Participants will be screened using the Monash machine learning GDM screening tool (MMLGDST). During the intervention (Block B), women at risk of developing GDM will be offered access to a smartphone-based coaching application featuring 1:1 synchronous sessions, asynchronous text and video messaging along with a Bluetooth-enabled weighing scale for self-monitoring. Support continues from early pregnancy through to nine months postpartum. The studys primary objective is to evaluate reach, adoption, implementation and maintenance of the B2B&Me+ intervention programme when delivered within routine maternity care. Implementation success will be assessed using the RE-AIM framework, while secondary outcomes will assess intervention effectiveness. The study will examine population-level uptake at each site, evaluate the benefits and costs of implementation across varying contexts, and analyse how four different referral methods, randomised at the site level, affect uptake. In addition, a European implementation toolkit will be developed to provide health services with scalable strategies to bridge the evidence-to-practice gap. Discussion This study will contribute to a growing literature on the implementation of a successfully trialled mHealth intervention in a real-world context. Understanding variation in both intervention and implementation success within a routine maternity care context across diverse settings will inform the development of an implementation toolkit to support health services in reducing the incidence of GDM and improving maternal and child health outcomes.

Background: Monitoring urinary iodine concentration (UIC) after sustained exposure informs iodine delivery strategies. We aimed to compare endline UIC among women of reproductive age (WRA) and preschool children (PSC) receiving multiple micronutrient-fortified (MMF) vs. iodine-only bouillon, and to describe population iodine status at baseline and endline. Methods: This pre-specified secondary outcome analysis used Condiment Micronutrient Innovation Trial (CoMIT) data. Non-pregnant, non-lactating WRA (15-49 years) and PSC (2-5 years) were enrolled at the household level. Households received MMF or iodine-only bouillon for 9 months (38 weeks); both contained iodine (KIO3) at 30 ug/g. Baseline and endline UIC (ug/L) were measured by modified Sandell-Kolthoff reaction in spot urine samples. ANCOVA models, adjusted for baseline log-UIC and recruitment site, compared endline geometric mean UIC between trial arms. Median UIC at both timepoints was compared to WHO cut-offs (ug/L): <100 (insufficient), 100-199 (adequate), 200-299 (above requirements), and >=300 (excessive); the prevalence of UIC <50 ug/L was calculated. Results: Endline UIC data were available for 611 WRA and 630 PSC. Adjusted endline UIC did not differ by arm (geometric mean ratio [95% CI]: WRA 0.97 [0.86-1.10]; PSC 0.96 [0.84-1.10]). With arms combined, median UIC increased among WRA (100.5 to 124.6 ug/L) and PSC (109.6 to 136.9 ug/L), with fewer samples <50 ug/L at endline. Conclusions: Adjusted endline UIC did not differ between trial arms. After 9 months' use of iodine-fortified bouillon, median UIC remained adequate among WRA and PSC, with fewer samples <50 ug/L at endline.

The development of cost-effective, serum-free media is critical for scalable cultivated meat production. This study used high-throughput screening through a Design of Experiments (DoE) approach to develop an animal-free, serum-free medium (MMM1) specifically for the C2C12 murine myoblasts model cell line with applicability in cultivated meat research including for pet food. Low cost, food-grade inputs such as methylcellulose and spirulina extract resulted in significant cell growth improvements. The optimised MMM1 formulation containing low cost, food-grade inputs, achieved cumulative population doublings comparable to 10% (v/v) fetal bovine serum over four consecutive passages. Furthermore, MMM1 supported scalable cell expansion on commercially available dextran-based microcarriers (Cytodex-3) in both static and agitated conditions in spinner flasks, matching growth rates of serum-based controls. Finally, transitioning to a food-grade DMEM/F12 basal medium maintained cell proliferation equivalent to the pharmaceutical-grade DMEM/F12, but at a significantly lower cost, thus offering a viable strategy to substantially reduce biomanufacturing costs which is a critical challenge in cultivated meat production.

Introduction: Infant neurodevelopment is associated with maternal nutritional status, yet few studies have evaluated the effects of balanced energy-protein (BEP) supplementation during pregnancy and/or lactation. Methods: The Maternal Infant Nutrition Trial (NCT03668977) was a 2x2 factorial, household randomized, unblinded, efficacy trial conducted in Sarlahi District, Nepal. Pregnant women were randomized to daily fortified BEP supplementation or no supplementation, and those with a live birth were re-randomized after delivery to supplementation or not until 6-months postpartum. The first 100 infants in each of the four intervention groups were enrolled in a neurodevelopment substudy. At 6-months, these infants underwent a cognitive, motor, language, and socioemotional assessment using the Bayley Scales of Infant and Toddler Development, fourth edition. Results: Baseline characteristics were similar across the four trial groups. A significant interaction (p< 0.05) between pregnancy and lactation supplementation groups was observed in the language domain only. Infants born to mothers who received BEP supplementation during pregnancy did not differ in standard scores for cognitive (mean difference (MD): 2.0, 95% confidence interval (CI): -0.29, 4.29), motor (MD: 0.98, 95% CI: -1.41, 3.36), or socioemotional (MD: -2.35, 95% CI: -5.27, 0.57) domains compared to the pregnancy control group. Similarly, for these three domains, there were no significant differences among infants whose mothers received BEP in lactation period vs. not. Infants born to mothers supplemented in both pregnancy and lactation had significantly higher mean language standard scores of 2.12 (95% CI: 0.16, 4.08; p-interaction = 0.015), driven by the expressive communication subtest, compared to those with no BEP supplementation in either period. Conclusion: BEP supplementation during pregnancy and/or lactation was not associated with improvements in cognitive, motor, or socioemotional domains at 6 months; however, infants of women supplemented in pregnancy and lactation had significant improvements in the language domain. Evaluation at later ages is warranted.

Turbot (Scophthalmus maximus) is an important aquaculture species, but the genomic organization and expressed diversity of its antibody repertoire remain incompletely characterized. In this study, we annotated the immunoglobulin heavy chain (IGH) locus using the haplotype resolved fScoMax1.1 genome assembly, and we used this as a reference to profile the expressed turbot IgM, IgD and IgT repertoires in skin and spleen. The primary IGH locus was located on chromosome 19, spanned approximately 72 kb, and contained 25 IGHV genes, including 24 functional genes and one pseudogene, together with three IGHD, seven IGHJ and three IGHC genes corresponding to IgT, IgM and IgD. Comparison with the alternate fScoMax1.1 haplotype and a second turbot genome assembly showed conserved IGHD, IGHJ and IGHC content, whereas IGHV gene number differed among assemblies. High throughput 5RACE repertoire sequencing revealed isotype and tissue associated differences in expressed IGH diversity. IgM represented the dominant productive repertoire in both skin and spleen and showed the highest clonotypic diversity, particularly in spleen. IgD displayed an intermediate profile, whereas IgT was more enriched in skin and exhibited the strongest clonal restriction. IGHV subgroup usage was dominated by IGHV3 in IgM and IgD, whereas IgT showed a distinct profile characterized by preferential use of IGHV4, especially in skin. Gene level analysis further showed broad IGHV-IGHJ pairing in IgM and IgD, with preferential use IGHJ3 segment, while IgT sequences paired exclusively with IGHJT. Clonotype sharing between skin and spleen was isotype dependent, being strongest for IgT, intermediate for IgM, and negligible for IgD, suggesting that clonal expansion did not necessarily predict inter tissue trafficking. Together, these results provide a curated genomic and expressed repertoire framework for turbot IGH genes and reveal isotype specific organization of antibody diversity, with IgT displaying a particular repertoire pattern.