Gastrointestinal nematode infections significantly impact equine health and welfare, with rising anthelmintic resistance demanding alternative control strategies. Emerging evidence suggests that parasitic nematodes harbour distinct microbiomes, potentially influencing host-parasite dynamics and parasite survival. This study aimed to characterize and compare the microbiomes of equine gastrointestinal nematodes and their hosts, focusing on differences in composition, diversity, and core microbiota structure across different intestinal sites, nematode subfamilies, and sexes. Faecal and nematode samples were collected from equids (Equus caballus and Equus asinus) at slaughterhouses. DNA was extracted, and the V3-V4 regions of the 16S rRNA gene were amplified and sequenced using the Illumina iSeq 100 platform. Bioinformatic analyses were performed with QIIME2 and MicrobiomeAnalyst, and statistical comparisons employed PERMANOVA, LEfSe, and alpha and beta diversity metrics. Nematodes exhibited a distinct microbiome dominated by Firmicutes, Proteobacteria, Bacteroidota, Verrucomicrobiota, and Actinobacteriota, differing significantly from the faecal microbiota. Alpha diversity analyses revealed lower richness in nematodes, while beta diversity indicated distinct community structures (p = 0.007). Microbial composition varied by gastrointestinal site, nematode subfamily, and sex. Proteobacteria were consistently enriched in nematodes, particularly in the caecum. Core microbiome analysis identified exclusive nematode-associated taxa such as Fusobacterium, Mesorhizobium, and Mycoplasma. Equine gastrointestinal nematodes harbour independent and structured microbiomes, distinct from those of their hosts. These findings underscore the ecological specialization of nematodes and highlight the potential of targeting parasite-associated microbiota for novel control strategies.

Methionine supplementation during the suckling period of ewe lambs may modify DNA methylation in the germline (F0) during this critical window period for the neonate. This circumstance may promote the intergenerational transmission of epigenetic marks to the offspring (F1), thus altering the expression of specific genes and physiological traits throughout F1 life. To test this hypothesis, 27 newborn ewe lambs (F1) born from either ewes being supplemented 0.1 % methionine (dry matter basis) during the suckling period (F0-MET) or not supplemented (F0-CTRL) were assigned to two different groups (F1-CTRL, n = 13 and F1-MET, n = 14), being the dietary treatment of their mothers (F0) the only source of variation. Thus, all the F1 animals (both groups) were raised exactly in the same way along the whole life (including lactation). In this study, we determined differences in the global blood methylation patterns, biochemical profile, and metabolome of female offspring (F1). Our data showed that functional categories such as those related to developmental process and anatomical structure development were significantly enriched in the F1-MET ewe lambs due to differentially methylated regions of genes in these categories. These F1-MET ewes also presented lower live body weight (P < 0.05) and reduced lipomobilization and milk yield (P = 0.099) during the lactation period, together with increased PUFAs content (P = 0.075) in the milk fatty acid profile when compared to F1-CTRL. Increased levels of insulin (P = 0.031) and beta-hydroxybutyrate (P = 0.043), along with certain features of the metabolome profiles, revealed altered lipid metabolism when compared to F1-CTRL animals. As far as fecal microbiota is concerned, no significant differences were found in alpha and beta diversity between the two groups. However, DESeq2 analysis performed on OTU-filtered data revealed that Treponema (genus) and Spirochaetaceae (family), both from the Spirochaetes phylum, were reduced in the F1-MET group compared to the F1-CTRL group (log2FC=-3.96, p(adj) < 0.05). The results suggest that early methionine supplementation in F0 ewe lambs has an intergenerational impact on their F1 female offspring, with negative consequences on lipomobilization and milk production during F1 lactation.

Rapid and non-destructive assessment of milk quality is crucial to ensuring both nutritional value and food safety. In this study, we investigated the potential of visible and hyperspectral imaging as cost-effective and quick-response alternatives to conventional chemical analyses for characterizing key properties of cowś milk. A total of 52 milk samples were analysed to determine their biochemical composition (polyphenols, antioxidant capacity, and fatty acids) using spectrophotometer methods and standard gas-liquid and high-performance liquid chromatography (GLC/HPLC). Concurrently, visible (RGB) images were captured using a standard smartphone, and hyperspectral data were acquired in the near-infrared range. A comprehensive analytical framework, including eleven different machine learning algorithms, was employed to correlate imaging features with biochemical measurements. Analysis of visible images accurately distinguished between fresh samples and those stored for 12 days (100 percent accuracy) and achieved perfect discrimination between antibiotic-treated and untreated groups (100 percent accuracy). Moreover, image-derived features enabled perfect prediction of the polyphenols content and the antioxidant capacity using an XGBoost model. Hyperspectral imaging further achieved classification accuracies exceeding 95 percent for several individual fatty acids and 94.8 percent for treatment groups using a Random Forest model. These findings demonstrate that both visible and hyperspectral imaging, when coupled with machine learning, are powerful, non-invasive tools for the rapid assessment of milkś chemical and nutritional profiles, highlighting the strong potential of imaging-based approaches for milk quality assessment.

This study investigated the effect of three levels of Camelina sativa seeds on ewes' diet on rumen microbiota using 16S rRNA gene amplicon sequencing and biochemical assays, focusing on rumen fermentation parameters and carbohydrates, proteins, and fats metabolism. Forty-eight dairy ewes were assigned to four homogeneous groups based on the inclusion level of C. sativa seeds in the diet (0, 28, 51.3, and 74.6 g/kg DM; Control, CS6, CS11, and CS16, respectively). Rumen digesta were collected on the 60th day of the trial using an esophageal tube. Rumen fluid was analyzed for volatile fatty acids (VFAs) concentration and rumen enzymatic activity. In addition, rumen microbiota was characterized in both fluid and solid fractions. The acetic and propionic acid concentrations were higher (P < 0.001) in CS11 compared with Control and CS6. Iso-butyric, butyric, iso-valeric, valeric acid, and total VFAs concentrations were higher (P < 0.05) in CS11 than in Control. The acetic: propionic ratio was higher (P = 0.003) in Control than in CS11 and CS16. Amylase activity was lower (P < 0.001) in CS6. In rumen fluid, CS11 and CS16 had a lower -biodiversity than the Control, while composition also differed (P ≤ 0.05). Regarding the most notable results of the taxa relative abundance, as compared with CS11 and CS16, Ruminobacter and Succinimonas relative abundances were lower (P < 0.05) in CS6, while Fibrobacter, Methanobrevibacter, Saccharofermentans, and Lachnospiraceae ND3007 group relative abundances were higher (P < 0.05). Ruminococcus was higher (P < 0.001) in CS6 than in Control. Fibrobacter was higher (P < 0.05) in the Control than in CS16. Butyrivibrio and Pseudobutyrivibrio were higher (P < 0.001) in CS11 compared with Control and CS6. Selenomonas -relative abundance was higher (P < 0.001) in CS16 than in Control and CS6. Higher inclusion levels were associated with reduced relative abundance of fibrolytic taxa and increased abundance of microbial groups linked to carbohydrate fermentation and -fatty acid biohydrogenation, with responses differing between rumen fluid and solid fractions. The lower-fat inclusion of C. sativa seeds could be associated with a relative enrichment of fibrolytic bacteria, potentially linked to greater phenolic bioavailabi-lity, whereas higher inclusion levels are associated with shifts toward amylolytic and propionate-associated taxa.

This study examines the potential of modulating gut health parameters in piglets by varying zinc/copper ratios administered through specialty oxide sources. A total of 84 piglets were selected after weaning and divided into four experimental treatment groups; the trial lasted 28 days. During the initial phase (1–14 d), the positive control (PC) received 2500 ppm of zinc, provided as conventional zinc oxide. In parallel, three additional treatment groups were formed, in which copper (Cu) and zinc (Zn) were supplemented using potentiated zinc oxide (Pot-ZnO) and monovalent copper oxide (Cu2O), at both European and Non-European recommended inclusion levels: EU (120 ppm of Zn; 140 ppm of Cu), Non-EU+ (300 ppm of Zn; 200 ppm of Cu), and Non-EU− (300 ppm of Zn; 140 ppm of Cu). Lower Zn/Cu ratios characterized the second phase (15–28 d). Growth performance and fecal score were monitored throughout the trial. Blood samples were collected on days 1 and 14 from one subject per replicate to assess serum proinflammatory cytokines, immunoglobulins, and biomarkers of intestinal permeability. On day 28, one subject per replicate was slaughtered to obtain jejunal mucosa for the determination of jejunal secretory immunoglobulin A (sIgA) and alkaline phosphatase. Fecal samples collected on days 14 and 28 were used to analyze Zn and Cu content and to evaluate microbiota composition. A better fecal score was detected on day 4 in PC group compared to EU (p < 0.01). Serum immunoglobulin A increased in Non-EU− vs. PC group (p < 0.05) at day 14. Serum diamine oxidase decreased in the PC and Non-EU+ groups vs. Non-EU− group (p < 0.05) at day 14. sIgA increased in PC vs. Non-EU− group (p < 0.01) at day 28. Zn was higher in PC fecal samples (p < 0.01), whereas fecal Cu increased in EU and Non-EU+ treatments at day 14 (p < 0.05). Analysis of fecal microbiota performed at day 14 showed decreased. Observed, Shannon, and Simpson metrics in the Non-EU− group compared to the EU group (p < 0.05). Beta diversity highlighted a significant separation among groups at day 14 (p < 0.01). Differential abundance analysis revealed notable changes in genera composition among PC and EU groups at day 14 (p < 0.05). In conclusion, balanced Pot-ZnO and Cu2O administered at inclusion levels compliant with European levels of inclusion (EU) represent a valid strategy to enhance gut health of piglets during the first two weeks after weaning.

IntroductionCondensed tannins (CT) influence ruminal microbiota, feed digestibility, and methane emissions, yet their effects in goats are poorly understood.MethodsThis study evaluated the impact of dietary quebracho CT extract at 0%, 2%, 4%, or 6% of dry matter on the composition of the dairy goat ruminal microbiota with a two-times repeated 4 × 4 Latin square design. Bacterial, archaeal, fungal, and protozoan communities were analyzed at the end of each feeding period for relative abundance changes, and their relationship to methane production, nutrient digestibility and feed efficiency were also assessed.ResultsIncreasing CT levels reduced alpha- and beta-diversity, with the 6% CT diet showing the most pronounced decline. CT inclusion induced phylum-level shifts in fiber-degrading microbes, including inversion of the Firmicutes to Bacteroidota ratio. Prevotellaceae and Succiniclasticum, tolerant to CT, increased significantly (P < 0.05), in line with higher propionate and lower methane production. The proteolytic bacteria Anaerolineaceae and Synergistaceae decreased (P < 0.05), consistently with the reduced isobutyrate and isovalerate ruminal concentration and with the reduced urinary nitrogen excretion. Methanobrevibacter, a key methane producer, was reduced by dietary CT (P < 0.05). The overall fungal biodiversity was also significantly changed (P < 0.05); the fiber-degrading Liebetanzomyces decreased, while the tannin-degrading Aspergillus increased (P < 0.05). Concerning protozoa, Diplodinium was reduced (P < 0.05) and Polyplastron and Isotrichia were increased (P < 0.05) by dietary CT.DiscussionThese and other microbial abundance changes correlated with reduced methane emission, altered fiber and protein digestibility, and modified volatile fatty acid (VFA) profiles. This study shows that decreased nutrient degradability in the rumen due to higher dietary CT alters the goat rumen microbiota and clarifies microbial taxa changes in relation to the zootechnical outcomes, including reduced methane production.

Milk microbiota is a complex microbial ecosystem with implications for product quality, safety, and animal health. However, limited data exist on goat milk microbiota, particularly in local breeds. This study provides the first detailed characterization of the milk microbiota of Girgentana goats, a resilient Sicilian breed valued for high-quality dairy products. Illumina NovaSeq sequencing was used to analyze the 16S rRNA V3–V4 regions of 44 individual and 3 bulk milk samples. Briefly, 16S rRNA-gene sequencing produced a total of 8,135,944 high-quality reads, identifying 1134 operational taxonomic units (OTUs) across all individual samples. On average, each sample showed 864 OTUs with counts > 0. Alpha diversity metrics, based on richness estimators (Chao1: 948.1; ACE: 936.3) and diversity indices (Shannon: 4.06; Simpson: 0.95; Fisher: 118.5), indicated a heterogeneous community with both common and low-abundance taxa. Firmicutes (51%) and Proteobacteria (27%) were the predominant phyla, with Lactobacillaceae (54%) and Bifidobacteriaceae (22%) dominating at the family level. Notably, farm bulk milk profiles closely mirrored individual samples. These results establish a milk microbiota baseline for the Girgentana breed and offer valuable insights into microbial ecology in traditional dairy systems, supporting future comparisons across breeds and farming practices.

This study aimed to investigate the effect of dietary supplementation with a probiotic live yeast Saccharomyces cerevisiae on rumen microbial communities, enzymatic activities, volatile fatty acid concentrations, and histology in artificially reared lambs fed a diet rich in fermentable carbohydrates. After colostrum administration, forty-two Chios lambs were divided into two homogenous groups (n = 21); the control (C) and probiotic supplemented group (P) and were artificially reared until the 45th day (d) of age. From the 30th until the 106th d of lambs' age, both groups were fed alfalfa hay and barley grain-based concentrate mix ad libitum. Moreover, 100 g of concentrate including 0.1 g of Saccharomyces cerevisiae CNCM I-1077 live yeast (1010 CFU/g) was offered to P lambs to ensure a constant intake of the yeast. Although no difference was evident for the alpha-diversity, several interesting features were found for beta-diversity. Butyrivibrio, Pseudobutyrivibrio, and acetate producer Sphaerochaeta were significantly more abundant (P = 0.029, P = 0.029 and P = 0.008, respectively) in P- compared to C-fed lambs at 100 d, while population of lactate users were varying significantly in P- and C- fed lambs at 100 d [higher abundance of Megasphaera in C- (P = 0.001) and Anaerovibrio in P-fed lambs (P = 0.046), respectively]. Acidaminococcaceae and Clostridia UCG-014 were found in a lower abundance (P = 0.059 and P = 0.008, respectively) in P- compared to C-fed lambs at 100 d. Both ruminal pH and the activities of fibrolytic enzymes did not differ amongst the dietary treatments, while total volatile fatty acid concentrations were increased (P < 0.001) in P- compared to C-fed lambs at 100 d. As regards the serosa, the muscular layers, the intramural vasculature and innervation, our results showed no effect in rumen histology due to the dietary treatment, however, changes were evident in the mucosa where all constituent layers of the epithelium presented distinctly and with similar thickness, especially around papillae, the collagen and reticular fibers were more densely and evenly packed, and prekeratins and keratins delineated more distinctly epithelial pegs in P-fed when compared to C-fed lambs. The inclusion of live yeast S. cerevisiae CNCM I-1077 promoted rumen microbiota stability and limited extensive keratinization of the epithelium due to a highly fermentable carbohydrate diet formulation.

In the Alpine region of Austria, Italy, and Switzerland, transhumance is widespread and the production of local traditional dairy products during summer is important. Raw milk cheeses are produced according to traditional recipes, using hurdles as a technique to guarantee food safety. In the present study, we aim to provide an overview of S. aureus and its enterotoxins in raw milk cheeses, identify the key parameters responsible for the enterotoxin production, and identify ways to improve food safety. The results demonstrate that safe artisanal raw milk cheese production is achievable under elementary conditions by applying effective hurdles, including high scalding temperatures or thermization, quality starter cultures, and robust milk quality management. The hurdle index (HI), which we introduce in this paper, is a promising tool for assessing and improving safety in raw milk cheese production.

This study investigated the effects of yeast-derived nucleotides (YN) supplementation via drinking water on growth performance, intestinal morphology, barrier function, and cecal microbiota in broiler chickens. A total of 480 one-day-old male Ross 308 broilers were randomly assigned to three groups: control (CTR), and two treatment groups receiving YN at 90 g/100 kg (T1) or 180 g/100 kg (T2) in drinking water for 42 days. Growth performance parameters were recorded throughout the trial, and intestinal samples were collected on days 21 and 42 for histomorphological evaluation, gene expression analysis, and microbiota profiling using 16S rRNA gene sequencing.YN supplementation significantly improved body weight on day 21 and average daily gain during the grower phase in the T2 group. Carcass weight was also significantly increased in T2 at day 21. Intestinal morphology analysis revealed improved villus-to-crypt ratio on day 21 in both treatment groups, and gene expression analysis demonstrated upregulation of tight junction-related genes (ZO-1, OCLN, CLDN3) in T2, indicating enhanced intestinal barrier integrity. No differences were observed in mucous layer thickness or MUC2 expression.Microbiota analysis showed a significant increase in alpha diversity indices over time, consistent with normal microbial maturation. While T1 increased Shannon and Simpson indices compared to CTR, no significant changes were observed for T2. Beta diversity was not affected by treatment, although a clear temporal shift in microbial community composition was evident between days 21 and 42.In conclusion, administering yeast-derived nucleotides via drinking water improved early growth performance and intestinal health in broilers, particularly at higher inclusion levels. This mode of administration offers a practical, flexible alternative to feed-based supplementation, allowing targeted use during critical growth periods or under stress conditions. These findings support the functional role of YN as a dietary strategy to promote gut health and performance in poultry production.

Bovine mastitis is a major challenge in dairy farms. Since the agents commonly used for pre- and post-dipping can affect the udder health by modifying milk microbiota, alternative products are needed. This study aimed to evaluate the effect of the use of pre- and post-dipping formulations containing the fermented broth of Nisin A-producing Lactococcus cremoris FT27 strain (treated group, TR) on the abundance and biodiversity of milk microbiota as compared to iodine-based commercial disinfectants (control group, CTR) during a three-month trial. The experiment was conducted on 20 dairy cows, divided into two groups (CTR and TR) of 10 lactating cows each. Milk samples were collected from two selected healthy quarters of each cow at 3 time-points. Microbial communities were investigated by cultural and sequence-based methods, and analyzed through bioinformatic and statistical approaches. Clear differences in bacterial community composition were observed among groups, with higher species richness in TR, especially of Staphylococcus, Enterococcus, Lactococcus, and Streptococcus genera. The microbiota was dominated by Firmicutes, followed by Actinobacteriota, Proteobacteria, and Bacteroidota. Staphylococcaceae family was significantly higher in TR (p < 0.009), whereas Carnobacteriaceae, Mycobacteriaceae, and Pseudomonadaceae were significantly lower (p = 0.005, p = 0.001, and p = 0.040, respectively). CTR had considerably higher abundances of the genera Alkalibacterium (p = 0.011), Pseudomonas_E (p = 0.045), Corynebacterium (p = 0.004), and Alloiococcus (p = 0.004), and lower abundances of Staphylococcus (p < 0.009). Milk microbiota changed noticeably during the experimental period, regardless of treatment. A significant decrease was observed in both groups for Firmicutes_A phylum, with an increment in Actinobacteriota phylum, Propionibacteriaceae family, and Cutibacterium genus. Streptococcaceae significantly decreased in CTR (p = 0.013) and rose in TR (p = 0.001). Several differences were observed between the two groups during the experimental period. Streptococcus genus almost disappeared in CTR (p = 0.013), whereas it significantly increased in TR (p = 0.001). Three and twelve enriched groups were significantly identified respectively in CTR and TR using LEfSe. The use of Nisin A-based teat dip formulations could be linked to greater microbial diversity compared to commercial products. Despite the influence of seasonality, the experimental formulations maintained higher milk biodiversity, suggesting that lactic acid bacteria metabolites prevent alterations in the milk microbiota.

BACKGROUND:Genetically modified soybean is largely used in animal feed and its massive cultivation affects the environmental sustainability of livestock and the dependency for the import in the European market. OBJECTIVES:The aim of this study was to evaluate the partial substitution of soybean meal with an innovative common bean genotype (Phaseolus vulgaris lec-lpa) with reduced content of anti-nutritional factors on zootechnical performance, gut microbiota modulation and faecal minerals in post-weaning piglets. METHODS:Fourteen piglets were divided into a control group fed with a basal diet and a treatment group fed with a commercial diet in which 7.3% of soybean meal and 0.8% of soybean oil were replaced with 10% of P. vulgaris lec- lpa for 28 days. BW, ADG, ADFI and FCR were evaluated, and diarrhoea incidence was recorded. Evaluation of pH, nitrogen content, protein digestibility and mineral content was performed on faecal samples. Microbiota was analysed by rectal swabs samples. Blood serum metabolic profile was evaluated. RESULTS:The treatment group showed lower BW and ADG during the trial (p < 0.05), but the health status of the animals was preserved. The treatment group released lower levels of minerals in faeces when compared with the control group after 28 days (p < 0.05) suggesting a lower dispersion of faecal minerals in the environment. Significant Beta diversity index was observed at 14 and 28 days (p < 0.05). Roseburia and Butyricicoccus increased in treatment group at day 28 (p < 0.05). These genera are associated with SCFA production, contributing to the maintenance of intestinal integrity, promoting positive bacterial populations and limiting inflammatory phenomena. CONCLUSIONS:In conclusion, P. vulgaris lec- lpa could be a viable and sustainable alternative protein source to reduce the European protein gap, playing a potential role in microbiota modulation and faecal minerals release.

Weizmannia-based probiotics (formerly allocated to Bacillus) have been studied for their benefits on intestinal health in monogastric animals, but their effects on calf rearing and diarrhoea-related intestinal challenges remain unclear. This is the first study evaluating the effect of Weizmannia faecalis DSM 32016 (WF-32016) on immunity and gut microbiota in female Italian Holstein Friesian calves. The trial was based on 20 female Italian Holstein Friesian calves, monitored from birth to ensure only healthy animals were included. Calves were randomly assigned to control (CTR) or probiotic (PRO) groups, with PRO receiving WF-32016 (109 CFU/kg feed), individually housed until nine weeks, and received a diet of colostrum, milk replacer, and starter feed. Colostrum samples, collected from dams at 6 h and 12 h post-birth to assess immunoglobulins G content, showed IgG levels above 50 g/l. At end of trial (day 56), probiotic-fed calves showed significantly greater body weight (PRO: 75.55 +/- 4.55 kg; CTR: 67.90 +/- 6.09 kg; p <= 0.05), and improved faecal scores in the first two weeks (PRO: 0.49 +/- 0.84; CTR: 1.06 +/- 1.14; p <= 0.01). Blood analysis indicated significantly higher IgG levels (p < 0.01) and antioxidant capacity (p < 0.01) in probiotic-fed calves at day 56. 16S rRNA analysis revealed significant beta-diversity differences (p < 0.05) and shifts in Lactobacillaceae, Streptococcaceae, Lachnospiraceae, and key genera (p < 0.05). Functional predictions indicated a significant increase in the pentose phosphate pathway in PRO samples (p < 0.05). WF-32016 supplementation enhanced growth, gut health, and immunity, supporting its potential as a natural alternative to reduce antibiotic dependence.

Sixty Alpine dairy goats were classified as healthy (HEAL, n = 30) or infected (INFE, n = 30) based on bacteriological culture of udder-half samples collected 7 d before dry-off. At -61 d from kidding (DFK), goats were dried off and randomly allocated to 2 homogeneous groups either receiving no treatment (15 HEAL, 15 INFE) or being treated intramammarily with 250 mg of cefazolin per half-udder (15 HEAL, 15 INFE). Milk yield, composition, and SCC were monitored at -82, 17, 45, and 80 DFK, and blood samples were collected at -66, -56, -7, and 8 DFK to assess plasma analytes. Antibiotic administration at dry-off did not affect productive performances in the new lactation or plasma analyte trends. Regardless of udder health status, lactose decreased in late lactation despite stable yield, likely due to increased SCC and mammary permeability during involution. The INFE goats had higher SCC before dry-off and higher SCS at 45 DFK. Following dry-off, plasma urea decreased across all groups, reflecting dietary changes aimed at easing milk cessation. Glucose and nonesterified fatty acids (NEFA) remained stable during the dry period. The INFE goats showed elevated plasma cholesterol at -57 DFK, suggesting transient dyslipidemia due to IMI. After kidding, all goats showed peaks in NEFA, NEFA/albumin ratio, BHB, bilirubin, and glutamate oxaloacetate transaminase, indicating body reserve mobilization and hepatic stress. Concurrently, increases in haptoglobin and ceruloplasmin, and decreases in albumin/globulin ratio and paraoxonase, reflect an acute phase response. The INFE goats showed higher reactive oxygen metabolites and thiol concentrations between -57 and 8 DFK, and elevated ceruloplasmin at -7 and 8 DFK, indicating sustained systemic inflammation. Plasma analytes could serve as effective diagnostic tools to improve the detection of subclinical mastitis in dairy goats.

This study aimed to assess the impact of a 15-d administration of a polyphenol-rich extract from hazelnut skin (HS) on the incidence of neonatal calf diarrhea (NCD) and bronchopneumonia (BP) in Holstein heifers. Additionally, the study investigated whether the extract influenced blood derivatives of reactive oxygen metabolites (d-ROM), serum antioxidant capacity, fecal microbiota, growth rates, and severity of BP and NCD-related clinical parameters. In this randomized clinical trial, 80 healthy female dairy calves were allocated into 2 groups: a control group (CTRL, n = 40) and an HS-supplemented group (HS, n = 40), which received a milk replacer enriched with HS extract (5 g/d) from d 3 to d 18 of life. Daily clinical examinations were conducted for all calves until they reached 21 d of age. Additionally, thoracic ultrasonography (TUS) was performed at 21, 40, and 60 d of age, and weights were recorded at enrollment (d 3), 21 d, and 60 d. These repeated outcomes were analyzed using generalized estimating equations. Blood samples were collected from each calf at the beginning of the study and after HS (or control) administration for the evaluation of serum antioxidant capacity using the oxygen adsorbent test (OXY-Adsorbent) and for determining d-ROM. Additionally, rectal swabs were taken from all calves at 2, 10, 18, and 24 d of life for the analysis of fecal microbiota. The duration of diarrhea in the CTRL group was 22% higher than in HS (incidence rate ratio = 1.22; 95% CI: 1.03-1.45). Calves in the CTRL group had higher odds of elevated fecal scores (odds ratio = 1.23; 95% CI: 1.02-1.52). At 21 d of life, median OXY-Adsorbent values were 397.29 µmol (25th percentile [25°P] = 360.51; 75th percentile [75°P] = 426.30) in CTRL and 440.40 µmol (25°P: 400.97; 75°P: 479.14) in HS, with a significant difference between groups The ADG, fecal microbiota, d-ROM, BP, disease incidence, and all clinical parameters evaluated were not affected by HS administration. In conclusion, these findings suggest that the administration of HS resulted in a reduction in the duration of watery feces during NCD episodes and an improvement in serum antioxidant capacity in treated calves. However, HS supplementation did not demonstrate any effects on the overall clinical health issues associated with NCD, lung lesions identified via TUS, or weight gain. Future research on HS supplementation in calves should be conducted over a more extended administration period and include comprehensive cost-benefit analyses.

Intramammary dry-off treatment is widely considered an effective method for preventing and curing intramammary infection (IMI) in lactating cows; however, it is not commonly used in small ruminants like goats. Therefore, this study was designed to evaluate the effect of an approved cefazolin-based intramammary treatment on the milk microbiota of Alpine dairy goats during the dry and early lactation periods. Sixty goats were randomly selected based on bacteriological results and randomly allocated into the control group (CG) or the treatment group (TG). Cefazolin 250 mg (Cefovet A, Dopharma, Firenze, Italy) was administered to the TG group at dry-off, whereas the CG received no treatment. Pooled milk samples were collected at dry-off (T1; 52 samples), colostrum (T2; 46 samples), and 5-10 days in milk (T3; 55 samples) for bacteriological analysis, somatic cell count (SCC), and 16 S rRNA gene sequencing. SCC levels were initially high in both groups at T1 (TG: 1,588,000 cells/mL; CG: 1,629,000 cells/mL), which significantly decreased at T3 (TG: 148,000 cells/mL; CG: 153,000 cells/mL). Notably, the TG had fewer infected mammary glands than the CG at T3 (p = 0.0248), while no differences were found at T1 or T2. Despite the reduction in SCC and infection rates, cefazolin-based treatment did not significantly affect the alpha- and beta-diversity between the TG and CG. On the other hand, shifts in microbial composition, including fluctuations in Firmicutes, Proteobacteria, and Actinobacteria, were primarily due to the lactation stage rather than the treatment. Differential abundance analyses identified non-pathogenic genera, such as Acinetobacter, Bacteroides, and Paracoccus, that varied between groups at different timepoints. The study provided insights into the effects of cefazolin-based dry goat treatment on goat milk microbiota and its changes during the lactation cycle, demonstrating its potential to reduce SCC and mammary gland infections without significant alterations to the milk microbiota.

Essential oils (EO) are known for their antibacterial and anti-inflammatory properties and can be used as an alternative to reduce the reliance on antimicrobials in dairy cattle. While many studies have explored the beneficial properties of EO in vitro, their effects on milk quality and milk microbiota, when applied directly to the udder skin, remain relatively unknown. This study aimed to investigate the impact of Thymus capitatus essential oil (TCEO), known for its high antibacterial and antioxidant properties, on milk microbiota using 16S rRNA sequencing, the lipidomic profile via liquid chromatography-quadrupole time-of-flight mass spectrometry, udder skin microbiota, and inflammatory biomarkers of dairy cows at the end of lactation. Sixteen-quarters of 12 Holstein cows were selected, and TCEO was topically applied to the udder skin twice a day for 7 days. Milk was collected aseptically on days 0, 7, 21, and 28 before morning farm milking. The results showed no significant changes in microbiota composition after the EO treatment in alpha and beta diversity or taxonomical composition at the phylum and genus levels. TCEO induced limited changes in the milk lipidome, primarily affecting diacylglycerols at T21. The treatment did not affect inflammatory biomarkers, milk sensory properties, or quality. Our study is the first to demonstrate that a local application of 10% TCEO on cow’s quarters does not significantly alter milk quality or microbiota composition in milk and skin. More studies should be conducted to ensure the safe use of TCEO in dairy cows and explore its potential benefits on antibiotic-resistant bacteria as an alternative or support for antibiotic therapy.

Background & aims: Considerable interest has been recently given to the potential role of the gut-brain axis (GBA)-a two-way communication network between the gut microbiota and the central nervous system- in the pathogenesis of attention-deficit hyperactivity disorder (ADHD), suggesting the potential usefulness of probiotic and synbiotic supplementations. In light of the limited available evidence, synbiotic efficacy in ADHD children not taking medications should be clarified. This study aimed to investigate the efficacy of a synbiotic dietary supplementation on fatty acids levels as well as on microbiota composition, behaviour, cognition, and brain function in children with ADHD. Methods: A total of 41 drug-na & iuml;ve school-aged children diagnosed with ADHD were enrolled in a 3month randomised, double-blind, comparison-controlled clinical trial, receiving either a synbiotic mix (COMP group) or the same synbiotic mix enriched with an additional extract from pigmented corn (EXP group). Changes in levels of some specific short-chain and branched-chain fatty acids were considered as primary outcomes. Secondary outcome measures included gut microbiota profiling, Child Behaviour Checklist, Conners Parent Rating Scale-revised, computerised cognitive tasks, and haemodynamic response to a Go-NoGo task measured by fNIRS. Results: No superiority of the EXP synbiotic mix was observed. Analysis of fatty acids did not reveal any significant difference between groups. Children in the COMP group reported a slightly greater improvement than those in the EXP group in focused attention and in the haemodynamic response to a cognitive task. Conclusions: This study shows that pigmented corn extract does not enhance the effects of the synbiotic supplementation in ADHD children in terms of fatty acid production, microbiota composition, clinical, cognitive and neurophysiological measures. However, a synbiotic mix of probiotics plus prebiotic acacia fibre and cornstarch could have some promising effects on ADHD symptoms, which warrants further research. Future studies should also continue to explore the potential of fNIRS for monitoring the effects of interventions that target the GBA. Trial registration: ClinicalTrials.gov: NCT06005506. (c) 2024 The Author(s). Published by Elsevier Ltd on behalf of European Society for Clinical Nutrition and Metabolism. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/ licenses/by-nc-nd/4.0/).

Background/Objectives: In the Aosta Valley, the alpine grazing system integrates livestock production and land management. Valdostana breeding has adapted to this mountainous region, but the spread of Staphylococcus aureus within pastures may impact animal health. The aim of this study was to provide an overview of S. aureus genotypes associated with antimicrobial resistance (AMR) and virulence profiles in four dairy herds in the Aosta Valley from July 2022 to August 2023. Methods: A total of 468 composite milk samples were collected at three timepoints: T1 (pasture-livestock system), T2 (farm-livestock system), and T3 (pasture-livestock system). S. aureus isolates were characterized by antimicrobial susceptibility testing, ribosomal spacer (RS)-PCR, multilocus sequence typing (MLST), PCR analysis for 28 virulence genes and 6 AMR genes, and adlb-targeted real-time PCR. Results: RS-PCR analysis of 82 S. aureus strains revealed 12 genotypes (GT) in eight clusters (CL). The most prevalent variant was GTRI (61%), followed by GTB (15%). Resistance to penicillin was high (69%), with CLR strains showing 88% resistance, and 51% resistance to amoxicillin plus clavulanate. All strains were susceptible to cephalosporins and oxacillin. Macrolide resistance was low (4%), and multi-drug resistance was 6%. AMR gene presence corresponded with susceptibility, with blaZ detected in 94% of CLR strains. CLR strains also possessed genes for biofilm formation and virulence factors. Conclusions: This study highlights the presence of AMR and virulence factors in S. aureus strains from alpine grazing systems, underscoring the need for ongoing monitoring to mitigate risks to animal health.

Nutritional strategies that supply methyl donors, such as methionine in the early post-natal period of lambs could program feed efficiency, milk production and metabolism along the whole life due to persistent changes in epigenetic marks. To test this hypothesis, 34 newborn ewe lambs from the same flock were stratified and distributed into two groups with equal body weight distribution. The control group (F0-CTRL, n = 17) was fed ad libitum with a milk replacer. In contrast, the second group (F0-MET, n = 17) received the same milk replacer supplemented with 1 g methionine/kg DM. After weaning, all animals were raised the same way and fed ad libitum with a complete pelleted diet formulated according to nutrient requirements. At 45 days of age, blood was collected from the jugular vein for DNA isolation, and DNA methylation was analyzed by Reduced Representation Bisulfite Sequencing (RRBS) in ten animals [5 ewe lambs (F0-MET) vs. 5 ewe lambs (F0-CTRL), corresponding to 5 twin births, of which one lamb was assigned to each group]. The main results show that CPT1B, a gene involved in fatty acid catabolism, was differentially methylated in F0-MET lambs (45 days of life). In addition, these animals presented increased insulin and non-esterified fatty acid concentrations while cholesterol levels were reduced, most of these changes being persistent along the whole life. No differences were observed in milk production and feed efficiency during lactation. Still, the milk's fatty acid (FA)