Bile acid profiles and mRNA expression of bile acid-related genes in the liver of dairy cows with high versus normal body condition.

Bile acids (BA) play a crucial role not only in lipid digestion but also in the regulation of overall energy homeostasis, including glucose and lipid metabolism. The aim of this study was to investigate BA profiles and mRNA expression of BA-related genes in the liver of high versus normal body condition in dairy cows. We hypothesized that body condition and the transition from gestation to lactation affect hepatic BA concentrations as well as the mRNA abundance of BA-related receptors, regulatory enzymes, and transporters. Therefore, we analyzed BA in the liver as well as the mRNA abundance of BA-related synthesizing enzymes, transporters, and receptors in the liver during the transition period in cows with different body conditions around calving. In a previously established animal model, 38 German Holstein cows were divided into groups with high body condition score (BCS) (HBCS; n = 19) or normal BCS (NBCS; n = 19) based on BCS and backfat thickness (BFT). Cows were fed diets aimed at achieving the targeted differences in BCS and BFT (NBCS: BCS <3.5, BFT <1.2 cm; HBCS: BCS >3.75, BFT >1.4 cm) until they were dried off at wk 7 before parturition. Both groups were fed identical diets during the dry period and subsequent lactation. Liver biopsies were taken at wk -7, 1, 3, and 12 relative to parturition. For BA measurement, a targeted metabolomics approach with LC-ESI-MS/MS was used to analyze BA in the liver. The mRNA abundance of targeted genes related to BA-synthesizing enzymes, transporters, and receptors in the liver was analyzed using microfluidic quantitative PCR. In total, we could detect 14 BA in the liver: 6 primary and 8 secondary BA, with glycocholic acid (GCA) being the most abundant one. The increase of glycine-conjugated BA after parturition, in parallel to increasing serum glycine concentrations may originate from an enhanced mobilization of muscle protein to meet the high nutritional requirements in early lactating cows. Higher DMI in NBCS cows compared with HBCS cows was associated with higher liver BA concentrations such as GCA, deoxycholic acid (DCA), and cholic acid (CA). The mRNA abundance of BA-related enzymes measured herein suggests the dominance of the alternative signaling pathway in the liver of HBCS cows. Overall, BA profiles and BA metabolism in the liver depend on both, the body condition and lactation-induced effects in periparturient dairy cows.

Role of miRNAs in assisted reproductive technology.

Cellular proteins and the mRNAs that encode them are key factors in oocyte and sperm development, and the mechanisms that regulate their translation and degradation play an important role during early embryogenesis. There is abundant evidence that expression of microRNAs (miRNAs) is crucial for embryo development and are highly involved in regulating translation during oocyte and early embryo development. MiRNAs are a group of short (18-24 nucleotides) non-coding RNA molecules that regulate post-transcriptional gene silencing. The miRNAs are secreted outside the cell by embryos during preimplantation embryo development. Understanding regulatory mechanisms involving miRNAs during gametogenesis and embryogenesis will provide insights into molecular pathways active during gamete formation and early embryo development. This review summarizes recent findings regarding multiple roles of miRNAs in molecular signaling, plus their transport during gametogenesis and embryo preimplantation.

INVITED REVIEW: Impact of Maternal Health and Nutrition on the Microbiome and Immune Development of Neonatal Calves.

This comprehensive review highlights the intricate interplay between maternal factors and the co-development of the microbiome and immune system in neonatal calves. Based on human and mouse studies, multiple prenatal and postnatal factors influence this process by altering the host-associated microbiomes (gut, respiratory tract, skin), microbial colonization trajectories, and priming of the immune systems (mucosal and systemic). This review emphasizes the importance of early life exposure, highlighting postnatal factors that work in synergy with maternal factors in further finetuning the co-development of the neonatal microbiome and immunity. In cattle, there is a general lack of research to identify the maternal effect on the early colonization process of neonatal calves (gut, respiratory tract) and its impact on the priming of the immune system. Past studies have primarily investigated the maternal effects on the passive transfer of immunity at birth. The co-development process of the microbiome and immune system is vital for lifelong health and production in cattle. Therefore, comprehensive research beyond the traditional focus on passive immunity is an essential step in this endeavor. Calf microbiome research reports the colonization of diverse bacterial communities in newborns, which is affected by the colostrum feeding method immediately after birth. In contrast to human studies reporting a strong link between maternal and infant bacterial communities, there is a lack of evidence to clearly define cow-to-calf transmission in cattle. Maternal exposure has been shown to promote the colonization of beneficial bacteria in neonatal calves. Nonetheless, calf microbiome research lacks links to early development of the immune system. An in-depth understanding of the impact of maternal factors on microbiomes and immunity will improve the management of pregnant cows to raise immune-fit neonatal calves. It is essential to investigate the diverse effects of maternal health conditions and nutrition during pregnancy on the gut microbiome and immunity of neonatal calves through collaboration among researchers from diverse fields such as microbiology, immunology, nutrition, veterinary science, and epidemiology.

Essential oil supplementation in milk replacers: short- and long-term impacts on feed efficiency, the faecal microbiota and the plasma metabolome in dairy calves.

Early supplementation with oregano essential oil (EO) in milk replacer (MR) may improve growth, immune responses, the microbiota and the metabolome in dairy calves during pre-weaning and in adulthood. Sixteen female dairy calves (3 days of age) were divided in two groups (n = 8/group): the control group (no EO) and the EO group (0.23 ml of EO in MR during 45 days). After weaning, calves were kept in a feedlot and fed ad libitum. The animals were weighed, and blood and faecal samples were collected on days 3 (T0), 45 (T1) and 370 (T2) to measure the biochemical profile and characterise peripheral blood mononuclear cells (PBMCs; CD4+, CD8+, CD14+, CD21+ and WC1+), the metabolome and microbiota composition. The EO group only had greater average daily weight gain during the suckling (EO supplementation) period (P = 0.030). The EO group showed higher average CD14+ population (monocytes) values, a lower abundance of Ruminococcaceae UCG-014, Faecalibacterium, Blautia and Alloprevotella and increased abundances of Allistipes and Akkermansia. The modification of some metabolites in plasma, such as butyric acid, 3-indole-propionic acid and succinic acid, particularly at T1, are consistent with intestinal microbiota changes. The data suggest that early EO supplementation increases feed efficiency only during the suckling period with notable changes in the microbiota and plasma metabolome; however, not all of these changes can be considered desirable from a gut health point of view. Additional research studies is required to demonstrate that EOs are a viable natural alternative to antibiotics for improving calf growth performance and health.

Bile acid profiles and mRNA abundance of bile acid-related genes in adipose tissue of dairy cows with high versus normal body condition.

Besides their lipid-digestive role, bile acids (BA) influence overall energy homeostasis, such as glucose and lipid metabolism. We hypothesized that BA along with their receptors, regulatory enzymes, and transporters are present in subcutaneous adipose tissue (scAT). In addition, we hypothesized that their mRNA abundance varies with the body condition of dairy cows around calving. Therefore, we analyzed BA in serum and scAT as well as the mRNA abundance of BA -related enzymes, transporters, and receptors in scAT during the transition period in cows with different body conditions around calving. In a previously established animal model, 38 German Holstein cows were divided into either a high (HBCS; n = 19) or normal BCS (NBCS; n = 19) group based on their body condition score (BCS) and back fat thickness (BFT). Cows were fed different diets to achieve the targeted differences in BCS and BFT (NBCS: BCS <3.5, BFT <1.2 cm; HBCS: BCS >3.75, BFT >1.4 cm) until dry-off at 7 wk ante partum. During the dry period and subsequent lactation, both groups were fed the same diets regarding their demands. Using a targeted metabolomics approach via LC-ESI-MS /MS, BA were analyzed in serum and scAT at wk -7, 1, 3, and 12 relative to parturition. In serum, 15 BA (cholic acid (CA), chenodeoxycholic acid (CDCA), glycocholic acid (GCA), taurocholic acid (TCA), glycochenodeoxycholic acid (GCDCA), taurochenodeoxycholic acid (TCDCA), deoxycholic acid (DCA), lithocholic acid (LCA), glycodeoxycholic acid (GDCA), glycolithocholic acid (GLCA), taurodeoxycholic acid (TDCA), taurolithocholic acid (TLCA), ß-muricholic acid (MCA(b)), tauromuricholic acid (sum of α and ß) (TMCA (a+b)), glycoursodeoxycholic acid (GUDCA)) were observed, whereas in scAT 7 BA (CA, GCA, TCA, GCDCA, TCDCA, GDCA, TDCA) were detected. In serum and scAT samples, the primary BA CA and its conjugate GCA were predominantly detected. Increasing serum concentrations of CA, CDCA, TCA, GCA, GCDCA, DCA, and MCA(b) with the onset of lactation might be related to the increasing DMI after parturition. Furthermore, serum concentrations of CA, CDCA, GCA, DCA, GCDCA, TCA, LCA, and GDCA were lower in HBCS cows compared with NBCS cows, concomitant with increased lipolysis in HBCS cows. The correlation between CA in serum and scAT may point to the transport of CA across cell membranes. Overall, the findings of the present study suggest a potential role of BA in lipid metabolism depending on the body condition of periparturient dairy cows.

Assessment of mycotoxin sequestration efficacy in Saccharomyces cerevisiae by-products cultured in wheat bran and whey protein medium.

Mycotoxins are metabolic products of fungi found in feed for farm animals and pose a major threat to food safety due to their adverse health effects. The development of strategies to reduce their bioavailability is crucial. In this context, the cell wall components of Saccharomyces cerevisiae (YCW), especially ß-D-glucans and Mannan-oligosaccharide, have been recognized as potent mycotoxin binders. The objective of this research was to develop a novel culture medium to increase the biomass yield of S. cerevisiae and optimize cell disruption by stepwise physical lysis and hydrolytic preconditioning. This process resulted in a yield of approximately 56% reducing saccharides and 28.54% protein. Subsequently, the ß-glucan was extracted after cell wall sequestration. The isolated YCW and extracted ß-glucan were characterized both individually and synergistically to evaluate their antibacterial properties and analyze their Fourier transform infrared (FTIR) spectra. In vitro evaluation of antibacterial activity revealed that a concentration greater than 250 µg/mL of YCW-ß-glucan blend significantly inhibited the growth of Gram-negative bacteria. In addition, this blend showed good adsorption of various mycotoxins, including Aflatoxin B1, Ochratoxin A, and Zearalenone, the latter of which exhibited a remarkable adsorption rate of 80.85%. This study highlights the promising potential of a combination of YCW and ß-glucan as a robust strategy to address the pervasive problem of mycotoxin contamination in feed.

Effects of Dietary Fiber Level and Forage Particle Size on Growth, Nutrient Digestion, Ruminal Fermentation, and Behavior of Weaned Holstein Calves under Heat Stress.

This experiment was designed to investigate the effects of feeding diets with different fiber content and forage particle size on the performance, health, nutrient digestion, rumen fermentation, and behavioral and sorting activity of Holstein dairy calves kept under elevated environmental temperature. Sixty weaned Holstein female calves (age = 96.7 ± 7.62 days old; body weight = 82.4 ± 10.4 kg) were randomly assigned to one of 4 treatments arranged in a 2-by-2 factorial design in a 70-day experiment. Dietary forage content (moderate, 22.5%; or high, 40.0% on DM basis) and alfalfa hay particle size (short, 4.39 mm; or long, 7.22 mm as geometric mean) were the experimental factors, resulting in the following combinations: (1) high-fiber (HF) diets with forage-to-concentrate ratio of 40:60 and long particle-sized alfalfa hay (LPS; HF-LPS); (2) HF diets with short particle-sized alfalfa hay (SPS; HF-SPS); (3) moderate-fiber (MF) diets with forage-to-concentrate ratio of 22.5:77.5 with LPS (MF-LPS); and (4) MF diets with SPS (MF-SPS). The temperature-humidity index averaged 73.0 ± 1.86, indicating that weaned calves experienced a moderate extent of heat stress. Fiber level and AH particle size interacted and affected dry matter intake, with the greatest intake (4.83 kg/d) observed in MF-SPS-fed calves. Final body weight was greater in calves receiving MF vs. HF diets (164 vs. 152 kg; p < 0.01). Respiration rate decreased when SPS vs. LPS AH was included in HF but not MF diet. Lower rectal temperature was recorded in calves fed MF vs. HF diet. Digestibility of dry matter and crude protein was greater in calves fed MF than HF diets, resulting in lower ruminal pH (6.12 vs. 6.30; p = 0.03). Fiber digestibility was greater in calves fed SPS compared with those fed LPS alfalfa hay. Feeding HF compared with MF diet increased acetate but lowered propionate molar proportions. The inclusion of SPS vs. LPS alfalfa hay decreased lying time in HF diet (920 vs. 861 min; p < 0.01). Calves fed MF vs. HF diets spent less time eating but more time lying, which is likely indicative of better animal comfort. Dietary fiber level and forage particle size interacted and affected sorting against 19 mm particles, the extent of which was greater in HF-SPS diet. Overall, dietary fiber level had a stronger effect than forage particle size on the performance of weaned calves exposed to a moderate degree of heat stress as feeding MF vs. HF diet resulted in greater feed intake, final body weight, structural growth measures, nutrient digestion, as well as longer lying behavior. The inclusion of SPS alfalfa hay in MF diets increased feed consumption.

Starter protein content and supplemental soybean oil or hydrogenated palm fatty acids in Holstein dairy calves: growth performance, protozoa population, and nitrogen utilization efficiency.

The study hypothesized that a 24% protein starter diet supplemented with inert fat (palm fatty acids; PLFs) instead of triglyceride-rich soybean oil (SBO) would promote the growth of dairy calves. The effects of different protein contents (20 vs 24% CP) and fat sources (SBO vs PLF at 2.5% DM) on growth, digestibility, blood metabolites, rumen protozoa and urinary nitrogen in dairy calves were investigated. A total of 52 female dairy calves (mean age 3 days and BW 40.7 kg) were allocated to the following treatments in a completely randomized design: (1) 20% CP starter diet supplemented with SBO (20CP-SBO), (2) 20% CP starter diet supplemented with PLF (20CP-PLF), (3) 24% CP starter diet supplemented with SBO (24CP-SBO), and (4) 24% CP starter diet supplemented with PLF (24CP-PLF). Calves were weaned on day 53, and the study ended on day 73. The 24CP diets enhanced starter feed intake, average daily gain (ADG), withers height, hip width, organic matter (OM) digestibility, and preweaning blood glucose compared to the 20CP diets. In contrast, SBO supplementation reduced feed intake, ADG, withers height, OM and CP digestibility, and pre- and postweaning blood glucose and beta-hydroxybutyrate levels, while increasing preweaning aspartate aminotransferase, compared to PLF. SBO supplementation led to lower urinary excretion of purine derivatives and microbial protein synthesis postweaning, and a reduction in protozoa population both pre- and postweaning. Calves fed PLF had higher starter intake than those fed SBO, regardless of the diet being 20CP or 24CP. Calves fed the 24CP-PLF diet had the highest ADG, while calves fed the 20CP-SBO diet had the lowest ADG. Calves fed the 20CP-SBO diet had lower feed efficiency than calves fed the other diets during the preweaning period and throughout the experimental period. Calves fed the 24CP-PLF diet had higher hip heights at weaning and on day 73 than calves fed the other diets. Calves fed the 20CP-SBO diet had lower neutral detergent fiber digestibility than calves fed the other diets. Calves fed the 24CP-SBO diet had higher postweaning blood urea nitrogen concentration than calves fed the other diets. Feeding the 20CP-SBO diet to dairy calves decreased urinary allantoin excretion and rumen microbial protein synthesis but increased urinary nitrogen excretion during the preweaning period. Our results suggest that PLF is more suitable than SBO in calf diets, especially when calves fed 24% CP, possibly due to improved nutrient digestibility.

Ultrasound characterization of mammary gland development in heifer calves fed at two different levels until weaning.

Ultrasound technologies allow for a non-invasive assessment of mammary gland (MG) development, the differentiation between the tissue types of the MG, and the evaluation of changes in its composition. This study aimed to work out a detailed description of the different stages of MG development that are visually discernible by ultrasonography for providing a template to classify the different structures. With this basis, the qualitative categorization of the developmental stage, as well as further quantitative assessments via pixel densities in the structures of interest, should be facilitated. Ultrasonic images were acquired from all four quarters of 37 German Holstein heifer calves fed either at a high feeding level of milk replacer (MR; 14% solids) at 10 L/day (1.4 kg MR/day; HI, n = 18) or at a restrictive low level of 5.7 L/day (0.8 kg MR/day; RES, n = 19) until linear weaning from week 13 to 14 of life. Ultrasound MG scans were performed first in week 3 of life, fortnightly from week 8-16, and in week 20 of life, in standing position, of each quarter, using a B-mode ultrasound device equipped with a linear probe (18 MHz). The developmental stages of the mammary gland parenchyma (PAR), visible in ultrasound images, obtained over 20 weeks of life, were categorized, described, and drawn by hand. On this basis, a template for classifying the visible categories of mammary PAR development and its surrounding tissue (SURR), and for measuring their pixel brightness was created thus providing an ultrasonographic atlas of the developing bovine MG, describing 11 categories. The ultrasound images were further classified by PAR structure, and pixel brightness was measured in PAR and SURR by using ImageJ Fiji. The difference in pixel brightness between PAR and SURR, the delta (Δ) pixel value was calculated. With increasing age, higher atlas categories of PAR developmental stages were shown. Pixel values, i.e. the brightness of PAR, its SURR, and Δ pixel value changed with age but were neither affected by the feeding group nor by a group × time interaction. With progressing PAR development, its pixel brightness increased from week 10 to 20 of life, i.e., the PAR became more hyperechoic since it spread and grew into its SURR. The atlas can serve as a template for the categorization and qualitative assessment of MG structures and for the quantitative assessment of PAR's development by measuring pixel brightness. With our study, we could show the structural development in PAR as well as in SURR in MG simultaneously in early life and confirm the spreading of PAR into its SURR by ultrasound scanning.

Effects of nomadic grazing system and indoor concentrate feeding systems on performance, behavior, blood parameters, and meat quality of finishing lambs.

The objective of the study was to evaluate the effects of three production systems on growth performance, behavior, blood parameters, carcass characteristics, and meat quality. A total of 30 lambs (n = 10 lambs/treatment) were randomly assigned to three production systems that included rotational grazing (NG) and two different levels of concentrate (CON), one with medium (roughage/concentrate ratio 50:50% based on DM, MC) and one with high concentrate (roughage/concentrate ratio 30:70% based on DM, HC) during the 90-day fattening period. At the start of the experiment, all lambs averaged 90 ± 4 days of age (mean ± SD) and were slaughtered at an average of 180 ± 3 days (mean ± SD). CON-fed lambs had higher average daily gain and loin thickness than NG-fed lambs. The NG lambs spent more time eating, drinking, and standing, but less time resting and rumination than the CON-fed lambs. In addition, plasma lipid, ß-hydroxybutyrate, and urea levels were higher, while phosphorus levels were lower in NG-fed lambs than in CON-fed lambs. CON-fed lambs had better carcass yield, but gastrointestinal tract and rumen weights were lower than NG lambs. CON-fed lambs had higher pH values 0 h post mortem than the NG lambs; however, there was no effect of treatment on pH 24 h post mortem. The post-mortem color of the LD muscle of NG lambs had a higher lightness and yellowness index and a lower redness index than that of the LD muscle of CON-fed lambs. The results of this study showed that lambs fed CON had better carcass yield than lambs fed NG, although feed intake, feed conversion ratio (FCR), growth performance, carcass yield, and meat quality of lambs fed MC and HC were similar.

Effects of dietary supplementation with histidine and ß-alanine on blood plasma metabolome of broiler chickens at different ages.

Histidine is an essential amino acid for broiler chickens and a precursor for the dipeptides carnosine and anserine, but little information is available about its metabolism in modern, fast-growing broilers. We used untargeted metabolomics to investigate the metabolic changes caused by the use of different standardized ileal digestible His:Lys ratios in broiler diets with and without ß-alanine supplementation. A total of 2204 broilers were randomly divided into 96 pens of 23 birds each. The pens were divided into 16 blocks, each containing one pen for all six feeding groups (total of 16 pens per group). These feeding groups were fed three different His:Lys ratios (0.44, 0.54, and 0.64, respectively) without and with a combination of 0.5% ß-alanine supplementation. Five randomly selected chickens of one single randomly selected pen per feeding group were slaughtered on day 35 or 54, blood was collected from the neck vessel, and plasma was used for untargeted metabolomic analysis. Here we show that up to 56.0% of all metabolites analyzed were altered by age, whereas only 1.8% of metabolites were affected by the His:Lys ratio in the diet, and 1.5% by ß-alanine supplementation. Two-factor analysis and metabolic pathway analysis showed no interaction between the His:Lys ratio and ß-alanine supplementation. The effect of the His:Lys ratio in the diet was limited to histidine metabolism with a greater change in formiminoglutamate concentration. Supplementation of ß-alanine showed changes in metabolites of several metabolic pathways; increased concentrations of 3-aminoisobutyrate showed the only direct relationship to ß-alanine metabolism. The supplementation of ß-alanine indicated few effects on histidine metabolism. These results suggest that the supplements used had limited effects or interactions on both His and ß-alanine metabolism. In contrast, the birds' age has the strongest influence on the metabolome.

Stress Concepts and Applications in Various Matrices with a Focus on Hair Cortisol and Analytical Methods.

When studying stress in animals, it is important to understand the types of stress and their classification, and how to assess the stress levels in different animal species using different matrices accurately and precisely. The classification of stress types helps to distinguish between good stress (eustress) and bad stress (distress). Hence, first, it is crucial to assess the animal's level of stress in a non-intrusive manner and second to identify the type of stress that is best suited to its environment. Third, it is also important to analyze the obtained samples using a suitable method to increase the validity of stress hormone measurements. Therefore, in this review, we aim to: (1) explain the classification of stress, (2) discuss the wide range of body matrices (e.g., saliva, milk, hair, urine, feces, sweat, fins, etc.) that can be used as samples to evaluate stress levels, as well as their comparisons and limitations, and present the reliable matrices for measuring stress hormones with special emphasis on hair, (3) compare the analytical methods for measuring stress hormones after sample preparation. Despite some literature that does not include hair as a reliable matrix for evaluating stress levels, hair is one of the matrices for measuring long-term stress hormone accumulations. This review discusses some factors that influence the level of stress hormones in the hair. By understanding these issues, the scientific community will not only be able to improve the understanding of stress and biomarker evaluation but also suggest how to deal with the consequences of stress in future research.

Effects of different milk feeding levels and frequencies on performance of Holstein heifers during weaning and first lactation.

In the present study, the effects of milk feeding amount and feeding frequency (FF) on performance, blood metabolites, rumen fermentation, and performance of dairy heifers during the first artificial insemination (AI) and first lactation were investigated. The treatment group consisted of 48 female Holstein heifer calves (12/treatment) distributed in a 2 × 2 factorial arrangement with milk feeding [high = 8 L/d, HL or medium = 6 L/d, ML] and feeding frequency [2 × or 3 ×]. All calves were fed on a step-up/step-down method, weaned from milk at 56 d of age, and observed until 70 d of age. Heifer calves were kept uniform from 70 d of age until the first lactation to evaluate the long-term effects of the treatments. We observed no FF effect and no interaction between the level and FF on feed intake, average daily gain (ADG), feed efficiency (FE), body weight (BW), structural growth, blood metabolites, and rumen fermentation parameters. Calves fed HL milk had higher total dry matter intake (DMI), BW, and ADG during the pre- and post-weaning periods. During the preweaning period, HL-fed calves had higher serum insulin (trend) and glucose concentrations compared to ML-fed calves. Lower age at first AI was observed in calves fed HL rather than ML regardless of FF. Weight at first AI, milk yield (305 d), and energy-corrected milk (ECM-305 d) in the first lactation showed a trend toward higher values in HL-fed calves compared to ML-fed calves. Preweaning performance and calf maturity can be positively affected by feeding high amounts of milk at both FF.

MicroRNA profiling of subcutaneous adipose tissue in periparturient dairy cows at high or moderate body condition.

A growing body of evidence shows that microRNA (miRNA), play important roles in regulating adipose tissue (AT) physiology and function. The objective was to characterize the AT miRNA profile in over-conditioned (HBCS, n = 19) versus moderate-conditioned (MBCS, n = 19) periparturient dairy cows. Tail-head subcutaneous AT biopsied on d -49 and 21 relative to parturition were used for miRNA sequencing. The miR-486 was the most significant miRNA among the upregulated miRNA on d -49, which might be related to more pronounced changes in lipogenesis and altered insulin sensitivity in AT of HBCS cows at dry-off. Comparing HBCS to MBCS on d 21, 23 miRNA were downregulated and 20 were upregulated. The predicted targets of upregulated differentially expressed (DE)-miRNA on d 21 were enriched in different pathways, including pathways related to lysosomes and peroxisomes. The predicted targets of downregulated DE-miRNA on d 21 were enriched in various pathways, including epidermal growth factor receptor, insulin resistance, hypoxia-inducible factor 1 signaling pathway, and autophagy. The results showed that over-conditioning was associated with changes in SCAT miRNA profile mainly on d 21, of which most were downregulated. The enriched pathways may participate in over-conditioning-associated metabolic challenges during early lactation.

Effects of freestall vs. bedded pack housing on growth performance, health status, and behavioral responses of weaned heifers.

This research evaluates the effect of two housing types, freestalls vs. bedded-pack, on growth performance, health scoring, and behavior of weaned heifers. Twenty-four heifers with no previous experience with freestalls (12 heifers/ treatment) were randomly assigned to the two treatments (freestalls vs. bedded-pack) at d 80 ± 3 of age. Heifers had free access to total mixed ration (TMR) and water throughout the experiment. The TMR intake and health score were recorded daily from d 80 to 110 of age. Body weight and skeletal growth of heifers were measured at the beginning (d 80) and the end (d110) of the experiment. Feeding behaviors were recorded for a period of 22 h and at the same time. Growth performance (ADG, structural growth, and final body weight), overall DMI, and health assessment (fecal score, general appearance, and rectal temperature) were similar between treatments. Behavioral patterns of eating, drinking, standing, lying, and rumination were also similar between treatments. However, in freestall-housed heifers, the proportion of time spent lying inside the stall increased by 8.1%, and time spent lying outside the stall decreased by 9.44% from the beginning to the end of the study (from 80 to 110 d). In summary, our results indicate that weaned heifers showed similar growth performance, behavior, and health when housed in freestall and bedded-pack housing. However, introducing weaned heifers to freestall housing at an early age will prepare them for using the facility.

Changes in the lipidome of water buffalo milk during intramammary infection by non-aureus Staphylococci.

This study aimed to determine the lipidome of water buffalo milk with intramammary infection (IMI) by non-aureus staphylococci (NAS), also defined as coagulase-negative staphylococci, using an untargeted lipidomic approach. Non-aureus Staphylococci are the most frequently isolated pathogens from dairy water buffalo milk during mastitis. A total of 17 milk samples from quarters affected by NAS-IMI were collected, and the lipidome was determined by liquid chromatography-quadrupole time-of-flight mass spectrometry. The results were compared with the lipidome determined on samples collected from 16 healthy quarters. The study identified 1934 different lipids, which were classified into 15 classes. The abundance of 72 lipids changed in NAS-IMI milk compared to healthy quarters. Significant changes occurred primarily in the class of free fatty acids. The results of this study provided first-time insight into the lipidome of dairy water buffalo milk. Moreover, the present findings provide evidence that NAS-IMI induces changes in water buffalo milk's lipidome.

Developmental programming: prenatal and postnatal consequences of hyperthermia in dairy cows and calves.

With global warming, the incidence of heat stress in dairy cows is increasing in many countries. Temperatures outside the thermoneutral zone (heat stress) are one of the environmental factors with the greatest impact on milk production and reproductive performance of dairy cows. In addition to several biological mechanisms that may contribute to the effects of fetal programming, epigenetic modifications have also been investigated as possible mediators of the observed associations between maternal heat stress during late gestation and performance and health later in life. In utero programming of these offspring may coordinate changes in thermoregulation, mammary gland development, and milk production ability at different developmental stages. This review examines the effects of prenatal and postnatal hyperthermia on the developmental outcomes of dairy cows, as well as the physiological and molecular mechanisms that may be responsible for the negative phenotypic consequences of heat stress that persist throughout the neonatal and adult periods and may have multigenerational implications. The physiological and molecular mechanisms underlying the negative phenotypic consequences of heat stress are discussed. Research challenges in this area, future research recommendations, and therapeutic applications are also discussed. In summary, strategies to reduce heat stress during the dry period should consider not only the productivity of the pregnant cow but also the well-being of the newborn calf.

Effects of group housing and incremental hay supplementation in calf starters at different ages on growth performance, behavior, and health.

The present study examined the effects of age at group housing and age at incremental hay supplementation in calf starters from 7.5 to 15% (dry matter, DM) and their interaction on growth performance, behavior, health of dairy calves, and development of heifers through first breeding. A total of 64 calves (n = 16 calves/treatment, 8 male and 8 female) were randomly assigned to 4 treatments in a 2 × 2 factorial arrangement, with age at group housing (early = d 28 ± 2, EG vs. late = d 70 ± 2, LG; 4 calves per group) and age at incremental hay supplementation of calf starters from 7.5 to 15% of DM (early = d 42 ± 2 d, EH vs. late = d 77 ± 2, LH) as the main factors. All calves (female and male) were weaned at 63 days of age and observed until 90 days of age. Heifer calves were managed uniformly from 90 days of age until first calving to evaluate the long-term effects of treatment. No interactions were observed between age at group housing and age at incremental hay to calves on starter feed intake, performance, calf health and behavior, and heifer development through first breeding, which was contrary to our hypothesis. The age at which incremental hay supplementation was administered had no effect on starter feed intake, growth performance, or heifer development until first calving. When EG calves were compared with LG calves, nutrient intake (starter, total dry matter, metabolizable energy, neutral detergent fiber, starch, and crude protein), average daily gain, and final body weight increased. In addition, frequency of standing decreased and time and frequency of eating increased in EG calves compared to LG calves. Overall, early group housing leads to improved growth performance in dairy calves with no negative effects on calf health compared to late group housing.