Intramammary infections (IMI), especially in their subclinical form, are a major concern in dairy herds due to their impact on udder health and productivity. This study assessed the associations between IMI induced by Staphylococcus aureus and 33 hematobiochemical parameters (inflammation, liver function, metabolism, and oxidative stress) in Holstein cows without clinical signs of disease. Four hundred multiparous cows were selected from a commercial herd based on genetic (estimated breeding values for somatic cell count [SCC]) and phenotypic data (SCC and mastitis history), and classified as resistant (TOP, n = 200) or susceptible (BOTTOM, n = 200) to mastitis. At time 0 (T0), milk cultures identified 37 cows positive for IMI pathogens (27 for S. aureus, 4 for S. dysgalactiae, and 3 for S. uberis) that were bacteriologically monitored also at T1 and T2 (2 and 4 weeks after T0, respectively) and sampled for the hematochemical parameters. Statistical analyses focused on cows infected with S. aureus, the only group with adequate case numbers. Infected cows showed evidence of systemic inflammatory and immune responses, such as increase in serum amyloid A (T1; p = 0.0496) and neutrophil counts (T2; p = 0.0337). Markers of hepatic function were also altered, with higher gamma-glutamyl transferase (T2; p = 0.0125) and aspartate aminotransferase (T2; p = 0.0304) concentrations, suggesting liver involvement during infection. Beta-hydroxybutyrate (T1; p = 0.0172) levels were elevated in infected cows, reflecting energy imbalance. Overall, these findings underscored the value of blood biomarkers for the early detection of affected animals.

Integrating multiple data layers could offer a better understanding of the pathogenetic mechanisms behind the development of mastitis and improve the identification of robust biomarkers for its detection. Hence, in this work we integrated serum metabolomic data from 1H nuclear magnetic resonance (1H-NMR) with milk leucocytes subpopulations measured with flow cytometry and milk udder health traits in healthy cows (n = 15, NEG) and cows with spontaneous subclinical intramammary infection (sIMI) (n = 19). The Data Integration Analysis for Biomarker discovery using Latent Components (DIABLO) algorithm was used to combine these datasets and identify key features for sIMI detection. The predictive ability of the selected hub variables in discriminating between NEG/sIMI animals was then assessed using receiver operating characteristic (ROC) analysis. This approach revealed a strong correlation (r = .73) between serum metabolomic and milk leukocytes categories. Among the most informative features selected by DIABLO we observed an increased concentration of lactose, due to the altered permeability of the mammary gland and histidine, potentially modulating immune responses. Additionally, the decreased concentrations of ruminal fermentation-associated metabolites (e.g. acetone, methanol, ethanol and 3-Hydroxybutyrate) suggest, with the onset of inflammation, a shift in energy allocation from physiological processes towards immune response. The predictive performance of the selected metabolites ranged from moderate to good. Lactose emerged as the most promising biomarker, while allantoin demonstrated high sensitivity (0.93). These findings demonstrate the potential of combining immunological and metabolomic profiling across different biofluids for mastitis detection, though larger cohort is required for testing its application.HIGHLIGHTS Strong relationship highlighted between milk immune cells and serum metabolomic variables Moderate correlations between milk leucocytes and serum metabolomic variables, but not with specific immune cells Infected animals show higher lactose and histidine levels Infected animals have lower ruminal fermentation-associated metabolites

Abstract Background Beyond conventional transcriptome profiling, RNA-sequencing (RNA-Seq) enables the discovery of variants within expressed genes linked to complex traits such as mastitis resistance or susceptibility. In this study, RNA-Seq was performed on milk somatic cells from uninfected Holstein cows with no history of mastitis (Neg, n = 9) or with subclinical intramammary infections (sIMI) caused by Prototheca spp. (P+ , n = 11) or Streptococcus agalactiae (Sa+ , n = 11). The objective was to identify transcriptome-derived sequence variants detectable under specific microbiological conditions that may contribute to the modulation of host transcriptional responses. By integrating these transcript-derived variants with quantitative trait locus (QTL) annotations and enrichment analyses, we aimed to highlight genomic regions functionally associated with mastitis susceptibility or resilience. Results Using the CLC Genomic Workbench, a total of 306,440, 264,132, and 246,777 unique variants were detected in Neg, P+ , and Sa+ groups, respectively, with SNPs being more abundant than INDELs. High-impact variants were identified in immune-related genes (TNIP1, TNIP3, IL10RB, IL2RA, IL15RA), suggesting post-transcriptional modulation of immune and inflammatory responses under different microbiological conditions. Variants in non-coding regulatory regions indicate that transcriptional control may also contribute to host susceptibility. QTL enrichment using GALLO showed consistent associations with milk production traits, while clinical mastitis QTLs were specifically enriched in P+ , reflecting context-dependent regulation of immune responses. Conclusions These findings provide new insights into the molecular basis of host responses to intramammary infections and support the value of a transcriptome-driven approach for variant discovery. Variants identified in immune-related genes and regulatory regions suggest that both immune gene modulation and post-transcriptional regulation may contribute to susceptibility to sIMI. Upon further functional validation, these findings may inform precision breeding strategies to enhance mastitis resilience in dairy cattle.

Heat stress (HS) represents a major challenge in dairy cattle production, as it is detrimental to the cows' welfare, feed intake, and milk yield and composition. Occurring under high temperature and humidity and commonly quantified by the temperature-humidity index (THI), it disrupts thermoregulation through behavioral and metabolic changes, thereby compromising productivity. In this study, we used accelerometers and in-line near-infrared (NIR) sensors to monitor the effects of THI levels on feeding (eating time and rumination) and panting behaviors assessed in terms of the time expended on them, dry matter intake (DMI), milk yield, and milk composition (expressed in percentages of fat, protein, lactose and casein) in Holstein cows. We analyzed records collected from 1,454 Holstein cows on 4 farms in Italy between September 2023 and January 2025. Eating time, rumination, and panting were monitored daily using AfiCollar accelerometer-based wearable sensors (785,195 hourly records), while milk yield and composition were measured in-line and in real time with a NIR AfiLab system (258,715 records). Daily THI was calculated from NASA/POWER data, and its effects were evaluated using segmented generalized additive mixed models (GAMMs) with fixed effects of farm, parity, stage of lactation, and milk productivity class, and random effects of cow and test date. Increasing THI levels had a consistently negative impact across all monitored parameters, but the magnitude of these effects varied with lactation stage, parity, and production level. Eating time and rumination time progressively declined at THI above 72, with the most pronounced reductions observed between 72 and 77, depending on the specific cow group (i.e., farm, days in milk, parity and daily milk yield) and trait of concern. Panting time increased significantly with rising THI in a biphasic pattern. Depending on the cow group evaluated, the sharp increase began at THI values between 70 and 72, and panting reached a plateau at THI values between 83 and 84.5. DMI declined markedly once THI exceeded 71.5 to 76, indicating the onset of voluntary feed restriction to limit metabolic heat production, and then plateaued near THI 88 for DIM, parity and milk yield class and 89 for farm. Milk yield showed substantial sensitivity to increasing THI, with the overall relative reduction from the group-specific threshold between THI 72-74 to THI 92 ranging from 37.1% to 56.7% across cow groups. The percentages of milk fat, protein, lactose, and casein also significantly decreased as THI exceeded values between 71 and 74. However, the early reduction in milk fat, rather than being solely attributable to HS, may partly reflect seasonal patterns, such as the 'spring effect'. Notably, high-producing cows frequently exhibited heightened sensitivity to heat load, whereas differences across lactation stages depended on the trait evaluated. Overall, HS had a detrimental impact on the eating time and rumination behavior, panting time, DMI, and milk yield and composition of Holstein dairy cows. Harmful effects began at around THI 70 to 74, establishing these values as critical indicators for intervention. Continuous sensor-based monitoring facilitates early detection of HS, enabling timely implementation of management strategies to ensure cow welfare and sustain productivity under thermal stress.

Intensified dairy production has increased cows' metabolic stress, which is often masked by good coping mechanisms that impede early detection of subclinical disorders and fertility decline. In particular, inflammation and oxidative stress compromise immune function, increasing susceptibility to disease. Noninvasive milk Fourier-transform mid-infrared (MIR) spectroscopy offers a promising alternative to laborious blood profiling for monitoring metabolic health. This study investigated the associations between milk infrared-predicted blood biomarkers of stress resilience and the sources of variation, focusing particularly on breed-specific responses in relation to DIM, parity, and dairy system. Previously developed calibration equations for 16 blood metabolites related to hepatic function and damage, oxidative stress, inflammation, and innate immunity were applied to a large population database using a gradient boosting machine approach. This database comprised 2,175,148 test-day records that included Fourier-transform MIR milk spectral data, obtained from 250,071 dairy cows from 1,509 multibreed farms belonging to the Parmigiano Reggiano Protected Designation of Origin Consortium. Records were collected by the Breeders Association of Emilia Romagna (Reggio Emilia, Italy) during official milk recording and released by the Italian Breeders Association (Rome, Italy). This dataset was merged with a field dataset provided by the Parmigiano Reggiano Consortium, comprising information on feeding and farm characteristics, allowing us to classify the farms into 4 dairy systems using a nonhierarchical k-means clustering procedure: 2 traditional systems (1 located in the Apennines, 1 in the Po Plain), and 2 modern systems, 1 using and 1 not using TMR. The 16 predicted biomarkers were analyzed individually using mixed models that included fixed effects of DIM, parity, month of sampling, year, breed, dairy system, the interactions breed × DIM, breed × parity, and breed × dairy system, and random effects of herd and animal. Overall, population-level MIR-predicted blood biomarkers were consistent with expected physiological patterns of DIM and parity, capturing postpartum hepatic overload and innate immune activation followed by gradual immune-metabolic recovery. Breed-specific patterns were evident: Brown Swiss cows exhibited lower hepatic recovery postpartum, maintaining higher concentrations of aspartate aminotransferase and alkaline phosphatase throughout lactation. We found breed- and farm system-specific variations in oxidative stress biomarkers, total reactive oxygen metabolites (ROMt), and advanced oxidation protein products, with Reggiana cows in modern systems exhibiting the highest levels of ROMt. Antioxidant biomarkers, ferric-reducing antioxidant power, and total thiol groups were elevated in traditional systems, especially in the Reggiana. Inflammatory biomarkers, such as haptoglobin and ceruloplasmin, varied according to breed and parity. Holsteins had the lowest ceruloplasmin levels, Brown Swiss and Reggiana cows the highest, regardless of system. Notably, Reggiana exhibited increased haptoglobin in late lactation. Because all patterns were consistent with physiological responses previously observed on a small scale using gold standard measures, these findings support the use of MIR-predicted biomarkers for routine population-level monitoring, potentially integrated with existing milk quality recording systems, and highlight opportunities for breed-specific health strategies tailored to different dairy systems. The predicted biomarkers could also serve as novel phenotypes for selective breeding.

Soybean meal is the primary protein source in pig diets, but its production has been associated with deforestation, soil degradation, and loss of biodiversity. Arthrospira platensis (AP), a protein-rich cyanobacterium with a favorable amino acid profile, might represent a sustainable alternative. This study aimed to assess the impact of partial to full replacement of soybean meal with AP on gut morphology and blood biochemical parameters in growing-finishing pigs. A total of 88 barrows and gilts were assigned to one of the four isoenergetic, isoproteic, and isoaminoacidic diets: a control diet (100% soybean meal as a protein source) and experimental diets in which 33, 66%, or 100% of soybean meal were replaced with AP. Individual blood samples were collected at the start of the trial (91 days), and prior to slaughter (238 days), when pigs’ body weight averaged 41 ± 3 kg and 175 ± 6 kg, respectively. Ileum and colon tissues were sampled at the slaughterhouse for histological analysis and evaluation of gut morphology. Across dietary treatments, biochemical profiling indicated metabolic, oxidative, and inflammatory stability, with no alterations in liver or kidney-related parameters. Consistently, histological evaluation indicated that intestinal architecture was preserved across all the dietary treatments, even though ileal villus width and crypt depth tended to increase linearly with higher dietary inclusion of AP, and a similar trend was observed for ileal inflammation score. Despite reports in other species suggesting AP antioxidative and anti-inflammatory benefits, such effects were not evident in pigs under the conditions tested. Nonetheless, the absence of adverse effects on intestinal and systemic health supports the nutritional viability of AP as a complete replacement for soybean meal. These findings highlight its potential as a sustainable and safe protein source in swine production without compromising physiological status or gut health.

This study evaluated the influence of sire genetic merit for residual feed intake (RFI) on carcass and ham quality traits in heavy pigs raised under the restricted feeding conditions typical of Protected Designation of Origin dry-cured ham production. A total of 417 purebred C21 Goland pigs, offspring of 23 sires, were randomly assigned to ad libitum, restricted medium-protein, or restricted low-protein dietary treatments. Sire breeding values (EBV) for RFI were estimated using RFI records of ad libitum-fed progeny between 96 and 161 kg body weight. Sires were classified into three RFI groups: low EBV (LRFI), medium EBV (MRFI), and high EBV (HRFI). Effects of RFI sire groups were estimated on traits recorded on the restricted-fed progeny. Progeny of LRFI sires exhibited differences in carcass traits, including a significantly higher carcass weight gain (+0.03 kg/day, corresponding to +7.3%; p< 0.01) and increased green ham yield (+1.3%, p < 0.05), compared to those of HRFI sires. LRFI progeny showed no differences in ham fat depth, but exhibited improved subcutaneous fat quality, including higher saturation (increased stearic acid, reduced linoleic acid and polyunsaturated fatty acids) and lower iodine number, as well as firmer subcutaneous fat. No variation in ham weight loss during dry-curing was observed across RFI sire groups. These findings suggest that selection for improved RFI does not significantly compromise carcass and dry-cured ham quality in restricted-fed heavy pigs. Incorporating RFI into selection objectives for sire lines could therefore provide a viable strategy for balancing production efficiency and product quality in heavy pig systems. These findings apply to restricted-fed heavy pigs of the population studied, and potential genotype × feeding regime interactions may limit direct extrapolation to other genetic backgrounds or production systems.

Journal of Animal Science and Technology (JAST) is a peer-reviewed, open access journal publishing original research, review articles and notes in all fields of animal science. Topics covered by the journal include: genetics and breeding, physiology, nutrition of monogastric animals, nutrition of ruminants, animal products (milk, meat, eggs and their by-products) and their processing, grasslands and roughages, livestock environment, animal biotechnology, animal behavior and welfare.

Diffuse large B-cell lymphoma (DLBCL) is one of the most prevalent haematological malignancies in both humans and dogs, characterised in both species by significant clinical heterogeneity and limited prognostic predictability. With the introduction of next-generation sequencing (NGS) technologies in veterinary medicine over the past decade, researchers have begun to elucidate the molecular basis of canine DLBCL (cDLBCL); however, much of the clinical heterogeneity associated with this tumour remains unexplained. In this study, we performed whole genome sequencing on 10 cDLBCL cases, all treated with chemo-immunotherapy, which exhibited similar clinico-pathological features but markedly different outcomes. Cases were classified as "poor" or "good" responders based on whether their lymphoma-specific survival fell below or above the cohort's median. Protein-coding variants and copy number aberrations unique to poor or good responders revealed novel candidate genes not previously identified in cDLBCL studies, while splicing, untranslated regions, and intronic variants were detected in genes already known to be recurrently mutated. In conclusion, our investigation has broadened the spectrum of potentially pathogenic variants implicated in cDLBCL, though further studies with larger cohorts are necessary to validate these findings.

Alternative splicing events lead to different mRNA isoforms, potentially translated into proteins with altered structures or impaired functions, or both. This phenomenon may influence the resistance or susceptibility to diseases, such as mastitis. To explore this aspect, this study aims to analyze transcript expression, focusing on cases where multiple mRNA isoforms are present, in milk somatic cells from Holstein cattle affected by subclinical intramammary infection caused by Prototheca spp. (P+, n = 11) or Streptococcus agalactiae (Sa+, n = 11), compared with uninfected animals (Neg, n = 9). The RNA-sequencing data were analyzed using the CLC Genomics Workbench (23.0.5, Qiagen) with a large gap read mapping approach and Bos taurus ARS-UCD1.3 reference genome to identify the differentially expressed transcripts (DET) among the groups. In addition, a functional analysis of the identified DET, combined with the identification of functional variants within the expressed regions, was performed. The comparison P+ versus Neg revealed 27 annotated DET, 11 annotated DET with novel length, and 7 novel DET with no previously annotated associated gene or length. These DET mainly originated from immune-related genes involved in pathways strictly linked to the immune and inflammatory responses (i.e., antigen presentation pathway, MHC class II antigen presentation pathway, macrophage classical activation signaling pathway). In contrast, the Sa+ versus Neg comparison revealed a total of 26 DET, including 17 annotated transcripts, 8 annotated transcripts with a novel length, and 1 novel transcript from a nonannotated gene. In this case, a predominance of enriched pathways related to metabolism and detoxification processes was observed (i.e., FXR/RXR activation, xenobiotic metabolism general signaling pathway, glutathione-mediated detoxification). Functional variants were identified in regions overlapping DET encoded by KRT78, CSN1S1, and MYBPC1, which were downregulated in P+ when compared with Neg group and potentially related to mastitis resistance/susceptibility traits. The transcripts and associated functional variants identified in this study may contribute to a better understanding of bovine mastitis pathogenesis and development, providing useful insights for improving animal health and management strategies.

Understanding the relationship between microbial community alterations and disease can provide valuable insights for improving diagnostics, prevention, and treatment strategies. This study used 16S rRNA amplicon sequencing to investigate and compare microbial diversity in the milk and feces of Holstein cows with subclinical mastitis caused by Streptococcus agalactiae and Prototheca spp. with that of healthy cows. A bacteriological screening identified 50 Holstein cows reared in a commercial dairy farm and classified into 3 experimental groups: (1) animals negative at the bacteriological examination with no history of subclinical mastitis (HLTH; n = 16), (2) animals positive at the bacteriological examination for either Streptococcus agalactiae (STRP; n = 22) or Prototheca spp. (PRTH; n = 12). The milk microbiota showed significant pathogen-specific alterations, with increased Firmicutes in STRP cows (55.6%) and Cyanobacteria in PRTH cows (17.3%), compared with healthy cows (39.2% and 0.7%, respectively). Alpha diversity (observed amplicon sequence variants, Shannon, and evenness indices) was significantly lower in infected cows, confirming a microbial imbalance. Beta diversity analysis (PERMANOVA) revealed significant differences in microbial composition between healthy and infected cows, but no significant differences in fecal microbiota composition. Differential abundance analysis identified Streptococcus (log2fold change [FC] = 7.3) as the most enriched taxon in STRP cows and Cyanobacteria (log2FC = 8.9) as the most enriched in PRTH cows in milk matrix, while Macrococcus caseolyticus was significantly reduced in both infected groups (log2FC = -4.5). These findings suggest that subclinical mastitis leads to significant shifts in the milk microbiota but does not alter the fecal microbiome, supporting a localized rather than systemic microbial response. This study provides novel insights into the microbial dynamics of subclinical mastitis and potential biomarkers for disease monitoring.

Enteric methane emissions (EME) are the main source of anthropogenic GHG in the dairy industry. Certified food products, such as Parmigiano Reggiano cheese, are particularly interested in monitoring their environmental impact. To achieve this, a rapid, cost-effective method, applicable at the population level, is to estimate EME from predicted milk fatty acids obtained by milk mid-infrared spectroscopy. The aim of this study was to evaluate the variation in EME-derived traits according to dairy system and farm-level animal welfare indicators in farms belonging to the Parmigiano Reggiano Consortium. The final dataset used for the analysis integrated milk test-day records, mid-infrared spectra, and farm animal welfare assessment data, resulting in 1,256,135 records from 174,180 cows on 940 multibreed dairy farms. Dairy systems were identified by clustering farms according to their common characteristics and practices. These included 2 traditional systems (one from the Apennine region and the other prevalent in the Po Plain) and 2 modern systems (both prevalent in the Po Plain, one with and the other without the use of TMR). This study analyzed different EME phenotypes, including methane emissions per day, emissions relative to kilograms of DMI, and emissions per unit of production (corrected milk, fresh cheese, and cheese solids). The effect of animal welfare and dairy systems on individual methane emissions was evaluated using mixed models. Three groups of farm-level welfare indicators were considered: management practices, housing structure and equipment, and animal-based measures. These indicators are part of the national animal welfare evaluation conducted by the Italian National Reference Center for Animal Welfare (CReNBA, Brescia, Italy). The results showed that the dairy system and the interaction between dairy system and breed affected all traits. The general trend was that modern dairy systems produced more grams of methane per day than traditional systems, but their emissions per unit of production were lower. Breed-related EME variations across dairy systems were especially notable in Reggiana cattle. The overall effect of the animal welfare on EME traits was minor compared with the dairy system or breed effects. Across the 5 EME traits analyzed, only 2 showed significant associations with management (methane expressed in g/d and relative to kg of DMI) and one with animal-based measures (methane production in g/d), whereas no effects were observed for structure and equipment. In light of these findings, producers of high-quality foodstuffs, such as Parmigiano Reggiano cheese, may take into account the impact of different sources of variation in EME in the process of selecting a suitable production system. The results of this study could be helpful in improving farm-level management, as well as in the development and implementation of breeding programs aimed at reducing the environmental impact of dairy farming.

The definition of dietary models that meet the animal's nutritional needs, improve its physiological state, and preserve the microbiota equilibrium, along with the challenge of reducing environmental effects is essential for producing sustainable, high-quality milk suitable for dairy production. This study aimed to investigate whether a more sustainable diet, with a higher content of alfalfa hay but equal net energy, protein, and NDF levels, has an effect on rumen fluid, feces, and milk microbiota in Sarda sheep. Additionally, it aimed to determine if microbiota changes across different niches correlated with production and traits. Twenty-four animals were divided into 2 groups and fed different diets. Rumen fluid, feces, and milk samples were collected at the beginning (d 1, T0) and at the end of the trial (d 22, T1). Microbial communities were analyzed using Ion GeneStudio S5 for sequencing the V2, V3, V4, V6-V7, V8, and V9 regions of the 16S rRNA gene. Amplicon sequence variant clustering and taxonomic assignment were performed with QIIME2 v2023.9.2. Differential abundant taxa between the 2 experimental groups (H-alfalfa, fed the high-alfalfa content diet, and L-alfalfa, fed the low-alfalfa content diet) were assessed at the 2 time points using the ANCOM-BC plugin of QIIME2, along with the analysis of the diversity indices. Similarly, milk composition, milk coagulation properties and BW, DMI, production, and energy requirements for milk production traits were analyzed for each time point using a linear model with diet as fixed effect. Results confirmed an overall homogeneity of the samples at T0 for milk composition, milk coagulation properties, and energy requirements for milk production, except for DMI. However, no significant difference in DMI was observed at T1 between experimental groups. Regarding the microbiota composition, analysis revealed minor differences between H-alfalfa and L-alfalfa groups in taxa abundances for rumen fluid and feces, both at T0 and 1. In milk, 8 taxa resulted to be enriched at T1 in the treated group. However, all the affected taxa had a low abundance, and no significant correlation was observed with the phenotypic traits. Diets did not affect the microbial structure of rumen fluid and milk, whereas significant differences in beta diversity (Bray-Curtis and unweighted UniFrac) were observed in feces at T1. Five distinct milk taxa resulted positively correlated with 3 taxa in rumen fluid and 2 in feces, supporting the idea that host microbial niches form a network of interconnected communities, even within isolated environments. In addition, a positive correlation was observed between energy requirements for milk production and feces' Oscillospiraceae family, a taxon typical of healthy gut in ruminants. The higher content of alfalfa hay showed no highly significant effects on microbiota composition, milk composition, milk coagulation traits, and energy requirements for milk production. Similarities in chemical composition and energy content between diets likely masked qualitative differences in the ingredients and resulted in minor differences between diet groups. Nevertheless, no adverse effects of the high-alfalfa hay diet were observed, suggesting its higher value as an effective strategy, particularly advantageous from an environmental perspective.

This study assessed the impact of rumen ‘fluid not adapted’ (FNA) and of rumen ‘fluid adapted’ (FA) to three additives on in vitro kinetics of gas production and end products of fermentation. The experiment was performed according to a Latin Square Design of 4 cows and 4 experimental periods. The dry cows received a total mixed ration designed for lactating cows without (control) or with 1 g/d of one of three additives (allyl-sulphide, cinnamaldehyde, limonene) that were selected for their known effects on rumen fermentation. The collected rumen fluids (FNA and FA, respectively) were used as inoculum of 4 consecutive in vitro incubations (1 for each experimental period) adding or not adding 30 mg of one of the three pure compounds. The incubations were performed using a commercial equipment to evaluate the kinetics of gas production and collect the end products of fermentation. The results indicated that FA did not influence any fermentation parameters compared to FNA. However, when allyl-sulphide was added in vitro, the effects of this compound tended to be more pronounced with FA than with FNA. This experiment highlights that the three tested pure additives, which show activity on in vitro fermentations, can alter the in vitro activity of rumen fluid collected from cows fed with these compounds. Therefore, the administration of pure additives directly to the cows can influence the rumen microbial activity and the response of in vitro experiments.

This study evaluated the effects of different SID lysine levels on growth performance, feed costs, and nitrogen excretion of growing pigs raised for dry-cured ham production. Ninety-six Goland-C21 x Camborough-43 pigs (initial body weight, BW: 44.5 +/- 3.8 kg) were allocated to four dietary treatments (2 pens/treatment, 12 pigs/pen) with low (L: 6.9, 6.0, 4.2 g/kg), medium-low (ML: 7.8, 6.9, 5.2 g/kg), medium-high (MH: 8.3, 7.8, 6.0 g/kg), or high (H: 9.3, 8.3, 6.9 g/kg) SID lysine across the growing (90-118 days), early finishing (119-132 days), and late finishing (133-233, days) phases. Automated feeding stations recorded individual feed intake. Backfat (BF) thickness and BW were measured at five time points. Nitrogen excretion was estimated from intake, retention, and atmospheric losses. Data were analysed using the PROC MIXED procedure in SAS; the model included dietary treatment, sex, and their interaction as fixed effects and pen nested within treatment as a random effect. Compared with L pigs, ML pigs showed similar feed intake and final BF depth but achieved higher (p < .05) final BW, greater average daily gain, and improved gain-to-feed ratio, while reducing feed costs (1.63 vs 1.77 per kg BW gain) without increasing nitrogen excretion. Growth performance of ML pigs was comparable to that of MH and H pigs; however, ML diets resulted in the lowest feed costs and nitrogen excretion. These findings suggest that a medium-low SID lysine diet is sufficient for this pig population, effectively supporting growth performance while reducing feeding costs and environmental impact.

Background The protein sources in pig diets strongly rely on soybean meal, but its production has been associated with soil degradation, deforestation and loss of biodiversity. Microalga Spirulina can be a potentially more sustainable alternative to soybean meal, but comprehensive information about its use in growing pigs is still lacking. This study aimed to evaluate the effects of partial to full replacement of dietary soybean meal with Spirulina on the growth and carcass traits of growing pigs and on the chemical and physical attributes of the meat. Methods Eighty-eight pigs, gilts and barrows mixed together, with initial body weight of 52.4 ± 4.2 kg, were allotted into 4 isoenergetic, isoproteic, and isoaminoacidic dietary treatments, which included a conventional control diet based on cereals and soybean meal and one of 3 diets formulated by replacing nearly 33%, 66% or 100% soybean with Spirulina. Each treatment had 2 pens (11 pigs/pen), which were equipped with electronic feeders that were able to record individual feed intake. After 138 d on feed, at 174.9 ± 6.4 kg body weight, the pigs were slaughtered, and the carcass traits and meat quality parameters from loin samples were assessed. Results The palatability of feeds was not depressed in pigs fed Spirulina, even when the soybean was completely replaced by the microalga. The incorporation of Spirulina in the diets in place of soybean did not impair the growth rate or feed efficiency, irrespective of the extent of replacement. The carcass traits and yield of commercial cuts were comparable for all Spirulina-included compared with those of the soybean-based groups, and the same was found for the chemical and physical attributes of loin meat. Conclusion The results obtained at the herd and slaughter levels revealed that the replacement of soybean meal with Spirulina did not negatively affect the growth or carcass traits of growing pigs or the main attributes of meat. Therefore, this study provides, for the first time, insights into the technical possibility of switching growing pig feeding systems toward more environmentally sustainable diets by including a microalga originating from landless feed production systems, which does not result in soil degradation or loss of biodiversity.

Increasing consumer concerns underscore the importance of verifying the practices and origins of food, especially certified premium products. The aim of this study was to evaluate the ability of Fourier-transform mid-infrared (FT-MIR) spectroscopy to authenticate animal welfare parameters, farming practices, and dairy systems. Data on farm characteristics were obtained from the Parmigiano Reggiano Consortium in northern Italy. Animal welfare data were collected by trained veterinarians using the assessment protocol developed by the Italian National Reference Center for Animal Welfare (CReNBA), and bulk milk test-day data were obtained from the laboratory of the Breeders Association of the Emilia Romagna Region. A merged final dataset of 12,083 bulk FT-MIR spectra records from 949 farms was created. Using a nonhierarchical clustering approach, the farms were classified into 5 dairy systems: 2 traditional systems comprising farms located in either the Apennines or the Po Plain; 2 modern systems, one that used TMR and one did not; and one traditional dairy system comprising farms rearing local breeds. To evaluate the ability of bulk milk to capture differences in farming systems, we conducted an ANOVA on milk composition. The linear models included the following effects: season, dairy system, farm, and the interaction between dairy system and season. The effect of the dairy system was significant for all milk composition traits. A 10-iteration linear discriminant analysis was used to evaluate the discriminative ability of the spectra in classifying farming practices and dairy systems. The average results of the area under the receiver operating characteristic curve revealed good authentication performance for genetic type (0.98), housing system (0.91), and feeding system (0.89), and medium-low authentication performance for geographical area (0.70); poor results were obtained for the percentage of concentrate in the diet and animal welfare parameters (0.57-0.64). With regard to dairy systems, the best result was obtained when dairy systems were grouped into 2 simplified categories, traditional versus modern (0.89), instead of the 5 categories (0.87). The results of this study show that FT-MIR is a useful tool for authenticating farming practices and dairy systems, but not animal welfare as defined by CReNBA evaluation criteria. Our results show that infrared spectroscopy has the potential to authenticate dairy products and quality label certifications.

Sustainability of animal production requires reducing reliance on soybean meal by identifying viable alternative protein sources. Within the framework of the Italian Agritech National Research Center, seven Italian research groups collaborated to evaluate unconventional feed ingredients and their effects on animal performance and product quality. Alternative legume seeds (peas, chickpeas, faba bean, and lupins) can partially or completely replace soybean meal without impairing productivity, while enhancing product health value and shelf-life through bioactive compounds. Microalgae (Chlorella, Spirulina) improved carotenoid content, antioxidant activity, fatty acid profile, and cholesterol levels in poultry products, with limited effects in pigs. Insects supported optimal growth in fish at 25–30% inclusion, whereas maximum recommended levels are 15% in broilers and 24% in laying hens to sustain growth, egg production, and quality. Camelina by-products are suitable for poultry diets at up to 5–10%, beyond which performance declines. Whole-plant soybean silage, tef (Eragrostis tef), and triticale–lupin intercropping represent promising protein-rich resources for ruminants, provided diets maintain balanced protein-to-energy ratios, adequate fibre characteristics, and appropriate harvest timing under drought-prone conditions. Collectively, these findings highlight the potential of diverse protein sources to improve the sustainability of livestock systems while preserving productivity and enhancing the nutritional quality of animal-derived foods.

The metabolic capacity of swine caecum-derived Lactobacillus spp. to biotransform mycotoxins presents promising potential as a host-probiotic strategy to improve pig health and support host-targeted probiotic research. In the present study, Lactobacillus spp. isolated from the pig caecum were examined for their ability to detoxify fumonisin B1 (FB1) in vitro. Three experimental groups were established (i) Control 1 (C1: buffer + caecal chyme), (ii) Control 2 (C2: buffer + FB1), (iii) Experimental group (E: buffer + caecal chyme + FB1), each with 12 replicates per group (4 replicates per time point 0, 24, and 48 h). Quantitative polymerase chain reaction (qPCR) was used to determine bacterial abundance, while fumonisin B1 (FB1) and its hydrolyzed product (HFB1, Hydrolyzed Fumonisin B1), were quantified using liquid chromatography-mass spectrometry (LC-MS). Group E showed a significant increase in Lactobacillus spp. abundance (p < 0.001), indicating a selective microbial response to FB1 exposure. In contrast, total bacterial counts did not differ significantly between C1 and E (p = 0.35), suggesting that the proliferation of Lactobacillus was the main microbiological outcome supporting the host-probiotic hypothesis. Principal component analysis (PCA) revealed distinct microbial clustering, explaining 97.3% of the variance. Compared to C2, FB1 levels in group E were significantly reduced at 24 and 48 h, while HFB1 conversion rates increased from 47.1% to 56.5%. The study identified Lactobacillus pontis, Lactobacillus amylovorus, and Lactobacillus ultunensis as promising host-associated probiotics, with potential application as feed additives to mitigate mycotoxin effects in pigs. These findings warrant further in vivo validation.

This study investigated the partitioning of metabolisable energy (ME) and standardised ileal digestible (SID) lysine in 96 Goland-C21 × Camborough-43 pigs, raised from 32 to 196 kg of body weight (BW) and fed voluntarily diets containing low (L), medium-low (ML), medium-high (MH), and high (H) SID lysine levels. Diets were formulated to provide decreasing SID lysine concentrations with increasing ages: early (90–118 d), late growing (119–146 d), and finishing (147–251 d). Individual feed intake was recorded using automated feeders, while BW and P2 backfat thickness were measured at the start and end of each interval to estimate protein (Pd) and lipid (Ld) deposition. Maintenance ME (MEm) was calculated by subtracting the energy required for Pd and Ld from total ME intake and scaled to BW0.60. Marginal SID lysine intake was determined as the difference between intake and maintenance requirements, based on the InraPorc model. The efficiency of SID lysine utilisation was calculated as the ratio of lysine deposited in Pd to marginal SID lysine intake. The L treatment provided insufficient SID lysine, resulting in lower final BW (p = 0.004), Pd (p = 0.003), and Ld (p = 0.023) compared to ML. However, ML, MH, and H treatments showed no significant differences. MEm averaged 0.961 MJ/kg0.60 BW, and lysine utilisation efficiency peaked at 0.68 in L pigs, aligning with InraPorc predictions. These findings support the applicability of the InraPorc model to pigs above 140 kg and offer updated SID lysine requirements across growth intervals.