The performance of multi-trait genomic prediction was assessed by simulating phenotypic data with the publicly available information on the genomic relationship matrix for 9850 Japanese Black cattle utilizing eigen decomposition. Variance component estimation and genomic breeding value prediction were performed by analyzing simulated phenotypic data of two different traits with four simulation settings of varied heritability (0.1, 0.4, and 0.7), genetic correlation (0.6 and 0.95), and phenotypic correlation (0.4 and 0.8). A total of 1000, 3000, and 5000 animals were selected as training populations. The performance of the algorithm for proven and young was also examined using simulated and real phenotypic data. Core animals were chosen from the 9850 animals to minimize the genetic variance of non-core animals conditional on the core animals using the genomic relationship matrix. Five sets of core animals were provided, and those explaining 25%, 50%, 75%, 90%, and 99% of the genetic variance were first explained. The multi-trait model gave greater prediction accuracy than the single-trait model for both traits, and not only heritability and genetic correlation but phenotypic correlation affected the accuracy in the current settings. Core animals explaining 90% and 99% variance could yield results similar to those obtained by using the original genomic relationship matrix.
Environmental enrichment (EE) is essential for promoting welfare and reducing atypical behaviors in captive research animals, but we know little about effective EE strategies for llamas (Lama glama). This study evaluated the behavioral responses to EE items provided to llamas housed in a biomedical research facility in Brazil, where they are maintained as biomodels for nanobody production. Eight EE items-including feeding, physical, and sensory stimuli-were tested through a longitudinal observational design supported by video analysis and ethograms. Descriptive statistics, repeated-measures ANOVA, and cluster analysis (K-means, hierarchical clustering, and principal component analysis [PCA]) were used to characterize behavioral patterns. Llamas showed a marked most time spent with the PVC pipe filled with hay or elephant grass, averaging 3.9 daily interactions. ANOVA indicated significant differences among animals (p < 0.001), with large effect sizes. Cluster analysis revealed two distinct behavioral profiles, confirming individual variation and identifying one llama with a unique interaction pattern. Sensory and recreational items (tire, grooming brush, and empty perforated ball) elicited low interaction, whereas forage-containing items consistently ranked highest. These findings highlight the importance of forage-based enrichment and demonstrate the value of behavioral analytics for designing dynamic EE programs that enhance welfare in llamas used in biomedical research.
Nine male pigs, which average 44.8 ± 3.6 kg body weight, were surgically fitted with a T cannula in the terminal ileum of the small intestine. The animals were assigned to a triplicated 3 × 2 incomplete Latin square design, consisting of three treatments: a complete mixture of industrial amino acids (L-AAs), a single cell protein (SCP) biomass coproduct, and an nitrogen-free diet. The animals were fed twice a day, with 5 days of adaptation to the experimental diets, followed by 2 days of continuous collection of ileal digesta for 10 h. Titanium dioxide was included in all diets at 0.5% as an indigestibility marker, and the N-free diet was used to estimate basal endogenous ileal losses of amino acids (AA). The standardized ileal digestibility (SID) of crude protein was higher (p < 0.05) for L-AAs compared to SCP (101.3% vs. 73.4%). For the indispensable and dispensable AA, the L-AAs also presented higher SID (p < 0.05) to SCP. Specifically for L-tryptophan, the SID of SCP was 89.2%. These results highlight that L-AAs are highly digestible, with SID values close to 100%, whereas SCP demonstrates viability as a source of indispensable and dispensable AA in swine nutrition.
MicroRNAs (miRNAs) play pivotal roles in intramuscular fat (IMF) deposition and determine animal meat quality. The study aimed to investigate the effect of miR-130b on adipogenesis in porcine intramuscular preadipocytes (PIMPA). In the study, PIMPA were isolated from longissimus dorsi muscle and cultured until 85%-90% confluence. The cells then subjected to adipogenic induction and transient transfections with miR-130b mimic, miR-130b inhibitor, and their corresponding controls (mNC and iNC). Oil red O staining was performed to detect lipid accumulation. Differentially expressed (DE) mRNAs were screened by transcriptome sequencing, and enriched functional pathways were identified through GO and KEGG analysis. The results demonstrated that numerous DE mRNAs were identified between the miR-130b mimic and mNC groups, which were mainly enriched in apoptosis, autophagy, and AMPK signaling pathways. Mechanistically, miR-130b overexpression significantly increased Bax expression, while reducing caspase-3 and Bcl-2 expression as well as Bcl-2/Bax ratio. It also, and tended to increase cleaved caspase-3/caspase-3 ratio, thereby activating apoptosis and suppressing lipid accumulation. Furthermore, these above results can be significantly reversed by miR-130b inhibition. In conclusion, our findings highlight that miR-130b inhibits lipid accumulation by activating apoptosis via Bcl-2/Bax/caspase-3 signaling pathway in PIMPA, providing a reliable molecular target for improving animal meat quality and ameliorating human obesity.
Maintaining the genetic composition of the captive founders is one of the essential tasks in ex situ captive breeding programs. Although minimization of the average coancestry of the current animals is the most widely accepted genetic management procedure when relationships among the captive founders are unknown (MC method), it has become possible to estimate the relatedness of the founders owing to the development of genotyping techniques and population genetics theory. In this study, we propose a novel genetic management procedure for preserving the genetic composition of captive founders that is applicable when the relatedness among them can be accurately estimated. The expression of genetic drift derived in this study suggested that Nei's minimum distance between the captive founders and the current animals is a key indicator for the novel genetic management. Computer simulation assuming unspecified organism indicated that the novel management preserves the original allele frequencies more effectively than the MC method, especially when the population size is contracting, and keeps the expected heterozygosity at a higher level, especially when the population size is expanding. We also exemplify the practical application of the proposed method by computer simulation using demographic data of the Oriental stork raised in Japan.
Accurate evaluation of body growth is essential for heifer management, and three-dimensional (3D) imaging offers a promising approach for precise morphological assessment. This study aimed to quantify variations in the abdominal morphology of dairy heifers using 3D imaging and to evaluate the relationship between abdominal volume and body weight. Twenty-seven records from 19 Holstein heifers aged 3-18 months were imaged using a three-camera 3D system. The abdomen was subdivided into 12 anatomical subregions to assess regional width variations. Age-related changes were evaluated using linear regression models, and age-adjusted residuals from the models were analyzed using principal component analysis (PCA) to characterize morphological differences. Abdominal volume increased linearly with age and was strongly associated with body weight (R2 = 0.96). Despite similar abdominal volumes, substantial variation in regional morphology was observed. PCA of the age-adjusted residuals indicated that overall abdominal width, proportional differences between cranial and caudal regions, and dorsoventral distribution patterns were the primary contributors to morphological variations. These findings demonstrate that 3D-derived abdominal volume strongly reflects body weight in growing dairy heifers, while regional morphological analysis reveals pronounced animal-level variations, highlighting the potential of 3D imaging for characterizing developmental differences in dairy heifers.
Immunostimulatory CpG oligodeoxynucleotides (CpG-ODNs) are potent activators of innate immunity with promising applications in human and veterinary medicine. We previously developed carbonate apatite-based oligonucleotide particles (ODNcaps) for oral delivery, which are efficiently taken up by macrophages in jejunal Peyer's patches. To enhance macrophage-specific targeting, we introduced glycan modifications to ODNcaps, exploiting carbohydrate-binding receptors on macrophages. In this study, glycosylated ODNcaps were synthesized through N-acetylglucosamine (GlcNAc) conjugation and evaluated for cellular uptake and immunostimulatory activity in mouse peritoneal macrophages. Uptake efficiency was quantified using fluorescently labeled CpG-ODNs, and immune activation was assessed by IL-6 mRNA expression and IL-6 secretion. GlcNAc-modified ODNcaps showed markedly higher uptake and induced the strongest IL-6 responses at both transcriptional and protein levels compared with non-glycosylated controls. These results demonstrate that glycosylation is a simple yet effective strategy to improve macrophage targeting and immunostimulation by CpG-ODNs. Beyond murine models, this platform may hold promise for oral or mucosal immune modulation in livestock such as cattle, poultry, and swine, offering a novel approach to enhance antigen-presenting cell activation and strengthen mucosal immunity in animal production.
Taurine, a sulfur-containing amino acid derived from methionine or cysteine, plays key roles in bile acid conjugation, antioxidant defense, and lipid and glucose metabolism. However, its relationship with physiological and metabolic changes in Japanese Black cattle, especially during the middle fattening stage, when plasma taurine rises with increased concentrate feeding, remains unclear. This study aimed to clarify the association between plasma taurine concentration and metabolic as well as ruminal characteristics during the middle fattening stage in Japanese Black steers (Bos taurus). Among the 21 steers, the highest (TauH, n = 6) and lowest (TauL, n = 6) steers in plasma taurine concentration were selected for comparison and their growth performance, blood metabolites, and rumen fermentation profiles were compared. The TauH group showed higher concentrate intake, greater ruminal propionate production, superior growth performance, and elevated plasma insulin, cholesterol, phospholipid, and aspartic acid concentrations than the TauL group. KEGG Orthology analysis indicated that methionine biosynthesis was enhanced in the rumen microbiota of TauH steers, whereas cysteine-related pathways predominated in TauL animals. Plasma taurine concentration closely reflected concentrate-driven metabolic adaptation, indicating that it represents a practical biomarker for assessing nutritional status and refining feeding strategies to improve growth and metabolic efficiency in beef cattle.
Adaptation of laying hens to heat stress involves variables and multiple mechanisms, which are complex and interconnected by linear responses and interactions. Here, we compare the Pearson correlation method and canonical correlation to evaluate relationships between thermoregulatory, behavioral, and productive responses and thermal environment variables in laying hens reared in a semi-arid region of Brazil. A total of 270 lightweight Hy-Line White laying hens, 58 weeks old, with body weight of 1.60 ± 0.092 kg and egg production of 77.30% ± 3.62% were used. Simple Pearson correlation analysis showed fewer significant relationships than those identified by canonical correlation analysis. The results showed low to moderate canonical correlations (0.2576 ≤ rc ≤ 0.7449) between sets of indicators. Relationships between the thermal environment, thermoregulatory responses, and productive responses were significant (p ≤ 0.05), with the pair thermoregulatory responses × thermal environment presenting the highest correlation (rc = 0.7449; rc2 = 0.5548). Canonical correlation analysis is recommended to assess the behavior of complex relationships in laying hens. This multivariate approach provides a comprehensive understanding of linear and interactive relationships and captures interactions between climatic variables and thermoregulatory, behavioral, and productive responses.
High-producing dairy cattle experience profound metabolic transitions during the periparturient and periweaning periods that influence health and long-term productivity. In ruminants, feed intake and metabolic adaptation reflect coordinated interactions among rumen fermentation, hepatic oxidative feedback, and gastrointestinal hormones. This review summarizes evidence on ghrelin and the glucagon-like peptide (GLP) axis and organizes recent findings within a hierarchical framework linking rumen-derived substrates, intestinal nutrient sensing, and systemic endocrine responses. Ghrelin increases during negative energy balance in early lactation and appears to support metabolic mobilization, whereas its orexigenic effect is constrained by ruminant-specific intake control. Evidence also indicates that postruminal amino acid supply and fatty acid profile can modulate ghrelin secretion, highlighting the importance of the digestive site and nutrient type. The GLP axis complements this regulation. GLP-1 links postruminal nutrient-related signals with insulin dynamics and satiety, whereas GLP-2 is more closely related to intestinal growth and adaptation during developmental transitions, including weaning. Notably, improvements in intestinal development in early life do not always coincide with large or sustained changes in circulating GLP-2. Overall, viewing dairy nutrition through endocrine responses, alongside nutrient supply, provides a basis to interpret variable outcomes in transition cows and calves and refine feeding strategies across physiological stages.
The objectives of this study were to explore the effects of 25-hydroxyvitamin D3 on (1) colostrum yield and compositions, 25-hydroxyvitamin D3, minerals, and health of periparturient cows and (2) 25-hydroxyvitamin D3 and minerals status, growth performance, and health of their calves. Forty multiparous dry cows were assigned to either vitamin D3 group or 25-hydroxyvitamin D3 group. After calving, all their calves received colostrum from their own dams; calves were respectively assigned to one of four treatments: (1) calves from D-VD group were fed vitamin D3; (2) calves from D-VD group were fed 25-hydroxyvitamin D3; (3) calves from D-25D group were fed vitamin D3; (4) calves from D-25D group were fed 25-hydroxyvitamin D3. Feeding 25-hydroxyvitamin D3 significantly increased (p < 0.05) 25-hydroxyvitamin D3, calcium and phosphorus in serum and colostrum, serum antioxidant capacity and immunoglobulin G (IgG) of periparturient cows, and significantly increased (p < 0.05) serum 25-hydroxyvitamin D3, calcium, and phosphorus in calves at birth. Feeding 25-hydroxyvitamin D3 calves improved (p < 0.05) weaning weight, skeletal growth, and average daily gain together with increased antioxidant capacity and IgG. In conclusion, supplementing 25-hydroxyvitamin D3 to periparturient cows and their calves improved colostrum and health of cows and growth and health of their calves.
This study investigated the effects of orally administered sugars and casein, main components of whole milk, on plasma glucagon-like peptide 1 (GLP-1) concentration in preweaning calves. In Experiment 1, calves received water (CON), 2.37% (w/w) glucose (GLU), 2.37% galactose (GAL), or a mixed solution containing 2.37% each of glucose and galactose (GLU + GAL). In Experiment 2, calves received 4.5% (w/w) lactose (LAC), 3.0% casein (CAS), or a mixed solution containing lactose at 4.5% and casein at 3.0% (LAC + CAS). Because baseline plasma GLP-1 and α-amino nitrogen (AAN) concentrations varied greatly and correlated with postadministration mean concentrations, statistical analyses for these parameters included baseline values as covariates. In Experiment 1, glucose and insulin concentrations were higher in GLU and GLU + GAL than in CON (p < 0.001), whereas GLP-1 concentrations were lower in GAL than in other groups (p < 0.001). In Experiment 2, glucose and insulin concentrations were higher in LAC and LAC + CAS, AAN concentrations were higher in CAS and LAC + CAS, and GLP-1 concentrations were higher in LAC, CAS, and LAC + CAS compared with CON (p < 0.001). In conclusion, both lactose and casein promoted GLP-1 secretion in calves, although oral administration of glucose or galactose alone did not elicit a marked GLP-1 response.
Some uncultured Succinivibrionaceae have been suggested to be associated with lower methane production in the rumen of cows, but this remains underexplored. This study aimed to elucidate the phylogenetic and ecological characteristics of uncultured Succinivibrionaceae. Phylogenetic differences between uncultured Succinivibrionaceae and other Succinivibrionaceae species were determined. The uncultured Succinivibrionaceae-specific primer set was newly designed. Real-time PCR was performed on 96 rumen fluid samples obtained from 69 Holstein cows and 12 calves to quantify the abundances of uncultured Succinivibrionaceae, Succinivibrio dextrinosolvens, Ruminobacter amylophilus, and archaea, and their abundances were compared. The 16S rRNA gene sequences of uncultured Succinivibrionaceae clustered separately from those of known Succinivibrionaceae species and were identified as a distinct bacterial cluster (US cluster). The US was highly abundant in calves and lactating cows but was scarce in dry cows. The short-chain fatty acid ratio [(C2 + C4)/C3] negatively and positively correlated with the US and other known Succinivibrionaceae species, respectively. The archaea-to-bacteria ratio negatively correlated with the US but positively correlated with known Succinivibrionaceae species. These phylogenetic and ecological differences between the US and known Succinivibrionaceae species suggest that the US has a different niche and functional role in the rumen from those of known Succinivibrionaceae species.
This study evaluated the effects of increasing inclusion levels of low-protein dried distillers' grains with solubles (DDGS) in high-concentrate diets on nutrient intake, digestibility, nitrogen balance, and ingestive behavior of finishing lambs. Four Santa Inês crossbred, uncastrated male lambs (24.8 ± 1.0 kg body weight) were assigned to four dietary treatments in a 4 × 4 Latin square design. Treatments consisted of a control diet without DDGS and diets containing 200, 400, or 600 g/kg dry matter (DM) of low-protein DDGS. DM and TDN intakes showed quadratic responses (p < 0.05) to DDGS inclusion. Neutral detergent fiber intake increased linearly, whereas NFC intake decreased linearly (p < 0.05). Inclusion of DDGS did not affect (p > 0.05) apparent digestibility of DM, CP, or EE. However, NDF digestibility increased linearly by 4.99 g/kg per unit of DDGS added. Nitrogen retention, plasma urea nitrogen, and blood glucose were not affected. Rumination time increased linearly (p < 0.05). Low-protein DDGS can be included at 600 g/kg DM in finishing lamb diets. However, including 200 or 400 g/kg DM may result in negative associative effects for some variables. Overall, low-protein DDGS remains a valuable feed ingredient for substituting ground corn and soybean meal.
The aim was to evaluate the supplementation of weaned piglets using a water-soluble flavored vitamin-mineral mixture. Ninety weaned piglets were distributed in a randomized block design with three treatments: control (CON), no supplement during nursery; W5D, with supplementation for 5 days after weaning and another 5 days before the end of Phase 3 and beginning of Phase 4; and W10D, with supplementation during the first 10 days after weaning. The W10D showed higher daily weight gain (p = 0.047) in Phase 3 and higher water expenditure during Phase 3 and Phase 4 (p < 0.05). The W5D and W10D reduced superoxide dismutase activity in erythrocytes and malondialdehyde concentration in plasma, indicating a positive effect on redox status. Behavioral assessment showed that W10D piglets spent more time on the feeder in the first 24 h after weaning compared with other groups (p = 0.045). Incidence of diarrhea was lower in the W5D and W10D treatments during the last phase (p = 0.002) and lower for the W10D treatment throughout the entire experimental period (p < 0.001). In conclusion, supplementation of a flavored vitamin-mineral mixture via water during the first 10 days postweaning improves the performance of piglets, with beneficial effects on redox status, behavior, and a reduction in the incidence of diarrhea.
This study investigated the effects of high-concentrate (HC) feeding during the growing stage on growth allocation and endocrine dynamics in Japanese Black heifers. Six heifers were assigned to a HC (3.0% body weight [BW]/day) or control (CON: 1.5% BW/day) group from 5 to 10 months of age. Feed intake, BW, muscle, bone, fat depot and visceral organ weights, histological traits of longissimus thoracis (LT), serum metabolites, and growth hormone (GH) dynamics were evaluated. At 10 months, the HC group showed greater BW and total muscle weight, with 18 muscles being heavier. The radius and ulna weights increased, but BW-adjusted ratios of several load-bearing bones tended to decrease. Subcutaneous fat deposition increased. LT adipocyte diameter and fat contents were greater, while muscle fiber type composition was unchanged. Liver weight and γ-glutamyl transferase activity increased. GH pulsatile secretion was reduced, whereas pituitary GH-positive cells, circulating IGF-1, and insulin concentrations remained unchanged. These findings indicate that HC feeding promotes overall growth but induces nonuniform development across tissues, reflecting a shift in growth allocation and endocrine regulation. Nutritional conditions during the growing stage may therefore play a critical role in shaping tissue development and subsequent productivity.
This study investigated whether the inflammatory component levels in bovine milk measured on the day of mastitis diagnosis were associated with recovery outcomes 7 days later. Milk was collected on Day 0 (the initial day of mastitis diagnosis) and Day 7 from 40 quarters of 32 dairy cows diagnosed with spontaneous acute mastitis. The pH, somatic cell count (SCC), and concentrations of serum amyloid A (SAA), interleukin-8, lactoferrin (LF), and sodium in milk were measured on Day 0 to evaluate their association with SCC on Day 7. A positive correlation was identified between SCC on Day 7 and SAA, LF, and Na levels on Day 0. For receiver operating characteristic analysis, a log10SCC > 4.8 (70,000 cells/mL) on Day 7 was considered indicative of mastitis. The cutoff values for SAA and LF on Day 0 were 21.6 and 81 μg/mL, respectively, with sensitivities of 79.3% and 86.2%, specificities of 77.8% and 66.7%, and areas under the curve of 0.784 and 0.762, respectively. These results indicate that measuring SAA and LF concentrations in milk at the initial mastitis diagnosis may predict intramammary conditions 7 days later, potentially aiding in understanding the pathology of mastitis and informing treatment strategies.
The objective of this study was to evaluate the individual effects of cellulase and Lactobacillus plantarum, as well as their interaction effect on the fermentation quality, microbial composition, and rumen degradation characteristics of apple pomace (AP) and wheat bran mixed silage. The experiment used a 2 × 2 factorial design with two factors: cellulase and L. plantarum, each with two levels (added or not added). The results demonstrated that cellulase supplementation in AP and wheat bran mixed silage increased acetic acid and total organic acid (TOA) levels while decreasing pH. Cellulase addition reduced neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents, increased water-soluble carbohydrate concentration, but elevated dry matter (DM) loss rate. Regarding rumen degradation characteristics, cellulase addition enhanced the effective rumen degradability of DM but reduced the effective rumen degradability of NDF and ADF. L. plantarum inoculation increased lactic acid and TOA levels, reduced pH and DM loss, and promoted a higher effective rumen degradation rate of NDF and ADF. Combined supplementation with cellulase and L. plantarum enhanced the relative abundance of lactic acid bacteria and the content of LA, reduced pH, NDF, and ADF, and increased ruminal effective degradability of nutrients. In conclusion, the combined supplementation of cellulase and L. plantarum exhibited a partial synergistic effect. The combined supplementation of cellulase and L. plantarum exerted a partial synergistic effect, preventing the increase in DM loss rate and the decrease in ruminal effective degradability of ADF induced by cellulase alone.
To clarify the effects of pork lipid profiles on eating quality, we examined the relationship between sensory evaluation values and the lipid profiles of M. longissimus thoracis and subcutaneous fat. Pork loins with a broad range of intramuscular fat contents and lipid profiles were collected nationwide. In Test 1, correlations between lipid profiles and sensory evaluation values of aroma were investigated using 96 pork loins. In muscle and subcutaneous fat, sweet aroma and monounsaturated fatty acids (MUFA) exhibited a significant positive correlation. Off-flavor and polyunsaturated fatty acids (PUFA) also showed a significant positive correlation. The MUFA/PUFA (M/P) ratio was significantly positively or negatively correlated with sweet aroma or off-flavors, respectively. For most items, overall acceptability of aroma and overall acceptability were positively or negatively associated with MUFA content and M/P ratio or PUFA levels, respectively. In Test 2, 60 pork loins were classified into four levels-< 3, 3-4, 4-5, and ≥ 5-based on the M/P ratio of subcutaneous fat and their effects on eating quality were investigated. A greater M/P ratio intensified sweet aroma and overall aroma acceptability while reducing off-flavors. These results identify that the M/P ratio is an effective indicator of eating quality.