Effects of Sweet and Forge Sorghum Silages Compared to Maize Silage without Additional Grain Supplement on Lactation Performance and Digestibility of Lactating Dairy Cows.

This study investigated the effects of replacing maize silage (MZS) with high-sugar sorghum silage (HSS) or forage sorghum silage (FSS) without additional grain supplement in the diets of dairy cows on nutrient digestibility, milk composition, nitrogen (N) use, and rumen fermentation. Twenty-four Chinese Holstein cows (545 ± 42.8 kg; 21.41 ± 0.62 kg milk yield; 150 ± 5.6 days in milk) were randomly assigned to three dietary treatments (n = 8 cows/treatment). The cows were fed ad libitum total mixed rations containing (dry matter basis) either 40% MZS (MZS-based diet), 40% HSS (HSS-based diet), or 40% FSS (FSS-based diet). The study lasted for 42 days, with 14 days devoted to adaptation, 21 days to daily feed intake and milk production, and 7 days to the sampling of feed, refusals, feces, urine, and rumen fluid. Milk production was measured twice daily, and digestibility was estimated using the method of acid-insoluble ash. The data were analyzed using a one-way ANOVA in SPSS 22.0 according to a completely randomized design. Dietary treatments were used as fixed effects and cows as random effects. The results indicate that MZS and HSS had greater crude protein but less neutral detergent fiber (NDF), acid detergent fiber (ADF), acid detergent lignin (ADL), and a lower pH than FSS (p ≤ 0.04). High starch contents in MZS and water-soluble carbohydrate (WSC) contents in HSS were observed (p < 0.01). While the highest starch intake was observed for the MZS-based diet, the highest WSC intake was noted for the HSS-based diet, and the highest NDF, ADF, ADL intake was observed for the FSS-based diet (p ≤ 0.05). The diets, including MZS and HSS, had greater digestibility than that of FSS (p ≤ 0.03). Feeding MZS- and HSS-based diets increased the yield, fat, and protein content of the milk, as well as feed conversion efficiency (p ≤ 0.03). However, feeding the MZS- and HSS-based diets decreased the contents of milk urea N, urinary urea N, and urinary N excretion more than the FSS-based diet (p ≤ 0.05). The N use efficiency tended to increase relative to diets containing MZS and HSS compared with FSS (p = 0.06 and p = 0.09). Ruminal ammonia-N and pH were lower, but total volatile fatty acids, acetate, and propionate were higher in cows fed the HSS- and MZS-based diets compared to those fed the FSS-based diet (p ≤ 0.03). It appears as though replacing MZS with HSS in the diet of cows without additional grain supplements has no negative influence on feed intake, milk yield, N utilization, or ruminal fermentation.

Effect of Rumen-Protected L-Tryptophan or L-Ascorbic Acid on Plasma Metabolites and Milk Production Characteristics of Lactating Holstein Cows during Summer Conditions.

This study investigated the effects of rumen-protected L-tryptophan or L-ascorbic acid supplementation on the productivity of lactating Holstein cows during a high-temperature period. Thirty cows were assigned to three dietary groups: control (CON), treatment 1 (TRT 1; rumen-protected L-tryptophan, 20 g/cow/d), and treatment 2 (TRT 2; rumen-protected L-ascorbic acid, 20 g/cow/d). As the high-temperature period progressed, the decrease in milk yield and dry matter intake (DMI) in the TRT 1 and TRT 2 groups was lower than that in the CON group. The total protein level in the plasma of the TRT 1 group was higher than that in the CON group (p < 0.05). Milk melatonin concentration was higher in the TRT 1 group than in the CON and TRT 2 groups (p < 0.05). Thus, the present results indicate that rumen-protected L-tryptophan or L-ascorbic acid has positive effects in preventing declines in DMI and milk yield by reducing heat stress in Holstein cows. In particular, rumen-protected L-tryptophan is considered effective in increasing the melatonin concentration in milk.

Identification and characterization of long non-coding RNAs in mammary gland tissues of Chinese Holstein cows.

LncRNAs (Long non-coding RNA) is an RNA molecule with a length of more than 200 bp. LncRNAs can directly act on mRNA, thus affecting the expression of downstream target genes and proteins, and widely participate in many important physiological and pathological regulation processes of the body. In this study, RNA-Seq was performed to detect lncRNAs from mammary gland tissues of three Chinese Holstein cows, including three cows at 7 d before calving and the same three cows at 30 d postpartum (early lactation stage). A total of 1,905 novel lncRNAs were detected, 57.3% of the predicted lncRNAs are ≥ 500 bp and 612 lncRNAs are intronic lncRNAs. The exon number of lncRNAs ranged from 2 to 10. A total of 96 lncRNAs were significantly differentially expressed between two stages, of which 47 were upregulated and 49 were downregulated. Pathway analysis found that target genes were mainly concentrated on the ECM-receptor interaction, Jak-STAT signaling pathway, PI3K-Akt signaling pathway, and TGF-beta signaling pathway. This study revealed the expression profile and characteristics of lncRNAs in the mammary gland tissues of Holstein cows at non-lactation and early lactation periods, and provided a basis for studying the functions of lncRNAs in Holstein cows during different lactation periods.

The mammary gland of dairy cows is the main place of milk synthesis and secretion, and plays a vital role in the process of milk production. LncRNAs (Long non-coding RNAs) are a class of non-coding RNAs with a length greater than 200 bp and do not encode protein, which can regulate gene expression at the transcriptional, post-transcriptional and chromatin levels, with biological functions such as regulating cell proliferation, differentiation, and apoptosis. Relevant studies in humans and model animals have shown that lncRNAs participate in mammalian mammary gland development and lactation, but there are few studies on lncRNAs regulation of mammary gland development and lactation in dairy cows. Therefore, this study aims to reveal the potential role of lncRNAs in the mammary gland of dairy cows through screening, identification, and functional research of differentially expressed lncRNAs at different periods of mammary gland development (pregnancy and early lactation period). It provides a reference for the follow-up study on the regulatory mechanism of dairy cows' mammary gland health.

Identification and characterization of whole blood gene expression and splicing quantitative trait loci during early to mid-lactation of dairy cattle.

BACKGROUND: Characterization of regulatory variants (e.g., gene expression quantitative trait loci, eQTL; gene splicing QTL, sQTL) is crucial for biologically interpreting molecular mechanisms underlying loci associated with complex traits. However, regulatory variants in dairy cattle, particularly in specific biological contexts (e.g., distinct lactation stages), remain largely unknown. In this study, we explored regulatory variants in whole blood samples collected during early to mid-lactation (22-150 days after calving) of 101 Holstein cows and analyzed them to decipher the regulatory mechanisms underlying complex traits in dairy cattle. RESULTS: We identified 14,303 genes and 227,705 intron clusters expressed in the white blood cells of 101 cattle. The average heritability of gene expression and intron excision ratio explained by cis-SNPs is 0.28 ± 0.13 and 0.25 ± 0.13, respectively. We identified 23,485 SNP-gene expression pairs and 18,166 SNP-intron cluster pairs in dairy cattle during early to mid-lactation. Compared with the 2,380,457 cis-eQTLs reported to be present in blood in the Cattle Genotype-Tissue Expression atlas (CattleGTEx), only 6,114 cis-eQTLs (P < 0.05) were detected in the present study. By conducting colocalization analysis between cis-e/sQTL and the results of genome-wide association studies (GWAS) from four traits, we identified a cis-e/sQTL (rs109421300) of the DGAT1 gene that might be a key marker in early to mid-lactation for milk yield, fat yield, protein yield, and somatic cell score (PP4 > 0.6). Finally, transcriptome-wide association studies (TWAS) revealed certain genes (e.g., FAM83H and TBC1D17) whose expression in white blood cells was significantly (P < 0.05) associated with complex traits. CONCLUSIONS: This study investigated the genetic regulation of gene expression and alternative splicing in dairy cows during early to mid-lactation and provided new insights into the regulatory mechanisms underlying complex traits of economic importance.

Analysis of blood biochemistry and non-targeted metabolomics of endometritis in dairy cows.

The incidence of bovine endometritis, which has a negative impact on the reproduction of dairy cows, has been recently increasing. In this study, the differential markers and metabolites of healthy cows and cows with endometritis were analyzed by measuring blood biochemical indicators and immune factors using biochemical and enzyme-linked immunosorbent assay kits combined with nontargeted metabolomics. The LC-QTOF platform was used to evaluate the serum metabolomics of healthy cows and cows with endometritis after 21-27 days of calving. The results showed that glucose, free fatty acid, calcium, sodium, albumin, and alanine aminotransferase levels were significantly lower in the serum of cows with endometritis than in healthy cows (P < 0.05). However, the serum potassium, interleukin-1, interleukin-6, and tumor necrosis factor levels were significantly higher in cows with endometritis (P < 0.05). In addition, the serum metabolome data analysis of the two groups showed that the expression of 468 metabolites was significantly different (P < 0.05), of which 291 were upregulated and 177 were downregulated. These metabolites were involved in 78 metabolic pathways, including amino acid, nucleotide, carbohydrate, lipid, and vitamin metabolism pathways; signal transduction pathways, and other biological pathways. Taken together, negative energy balance and immune activation, which are related to local abnormalities in amino acid, lipid, and carbohydrate metabolism, were the important causes of endometritis in dairy cows. Metabolites such as glucose, carnosine, dehydroascorbic acid, L-malic acid, tetrahydrofolic acid, and UDP-glucose may be used as key indicators in the hematological diagnosis and treatment of endometritis in dairy cows.

Estimation of genetic parameters and single-step genome-wide association studies for eating time and rumination time in Holstein dairy cows.

The aims of this study were to estimate genetic parameters and to identify genomic regions associated with eating time (ET) and rumination time (RUT) in Holstein dairy cows. Genetic correlations among ET, RUT, and milk yield traits were also estimated. The data were collected from 2019 to 2022 in 6 dairy herds located in the Walloon Region of Belgium. The dataset consisted of daily ET and RUT records on 284 Holstein cows, from which 41 cows had records only for the first parity (P1), 101 cows had records from both the first and second parities, and 142 cows had records only for the second parity (P2). The number of daily ET and RUT records in the P1 and P2 cows were 18,569 (on 142 cows) and 34,464 (on 243 cows), respectively. Data on 28,994 SNPs located on 29 Bos taurus autosomes (BTA) of 747 animals (435 males) were used. Random regression test-day models were used to estimate genetic parameters through the Bayesian Gibbs sampling method. The SNP solutions were estimated using a single-step genomic best linear unbiased prediction approach. The proportion of genetic variance explained by each 20-SNP sliding window (with an average size of 1.52 Mb) was calculated, and regions accounting for at least 1.0% of the total additive genetic variance were used to search for candidate genes. Mean (standard deviation; SD) averaged daily ET and RUT were 327.0 (85.66) and 559.4 (77.69) min/d for cows in P1 and 316.0 (82.24) and 574.2 (75.42) min/d for cows in P2, respectively. Mean (standard deviation; SD) heritability estimates for daily ET and RUT were 0.42 (0.09) and 0.45 (0.06) for cows in P1 and 0.45 (0.04) and 0.43 (0.02) for cows in P2, respectively. Mean (SD) daily genetic correlations between daily ET and RUT were 0.27 (0.07) for P1 and 0.34 (0.08) for P2. Genome-wide association analyses identified 6 genomic regions distributed over 5 chromosomes (BTA1, BTA4, BTA11, 2 regions of BTA14, and BTA17) associated with ET or RUT. The findings of this study increase our preliminary understanding of the genetic background of feeding behavior in dairy cows; however, larger datasets are needed to determine whether ET and RUT might have the potential to be used in selection programs.

Altered rumen microbiome and correlations of the metabolome in heat-stressed dairy cows at different growth stages.

IMPORTANCE: Heat stress is one of the main causes of economic losses in the dairy industry worldwide; however, the mechanisms associated with the metabolic and microbial changes in heat stress remain unclear. Here, we characterized both the changes in metabolites, rumen microbial communities, and their functional potential indices derived from rumen fluid and serum samples from cows at different growth stages and under different climates. This study highlights that the rumen microbe may be involved in the regulation of lipid metabolism by modulating the fatty acyl metabolites. Under heat stress, the changes in the metabolic status of growing heifers, heifers, and lactating cows were closely related to arachidonic acid metabolism, fatty acid biosynthesis, and energy metabolism. Moreover, this study provides new markers for further research to understand the effects of heat stress on the physiological metabolism of Holstein cows and the time-dependent changes associated with growth stages.

Identification and Classification of Long Non-Coding RNAs in the Mammary Gland of the Holstein Cow.

The mammary glands, responsible for milk secretion, are regulated at a local level by various hormones, growth factors, non-coding RNAs, and other elements. Recent research has discovered the presence of lncRNAs in these glands, with suggestions that they may be essential for the maintenance and function of mammary glands. Besides directly controlling the gene and protein expression, lncRNAs are believed to play a significant part in numerous physiological and pathological processes. This study focused on examining the mammary gland tissues of Chinese Holstein cows, to identify and categorize long non-coding RNAs (lncRNAs). The research intended to distinguish lncRNAs in the mammary tissues of Holstein cows and contrast them between lactation and non-lactation periods. In this study, mammary gland tissues were sampled from three Holstein cows in early lactation (n = 3, 30 days postpartum) and non-lactation (n = 3, 315 days postpartum) on a large dairy farm in Jiangsu province. Mammary tissue samples were collected during early lactation and again during non-lactation. In total, we detected 1905 lncRNAs, with 57.3% being 500 bp and 612 intronic lncRNAs. The exon count for lncRNAs varied from 2 to 10. It was observed that 96 lncRNA expressions markedly differed between the two stages, with 83 genes being upregulated and 53 downregulated. Enrichment analysis results revealed that Gene Ontology (GO) analysis was primarily abundant in cellular processes. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that target genes were predominantly abundant in metabolic pathways, fatty acid biosynthesis, the immune system, and glycosphingolipid biosynthesis. This study analyzed the expression profile and characteristics of lncRNAs in the mammary gland tissues of Holstein cows during both lactation and non-lactation stages, forming a foundation for further investigation into the functional roles of lncRNAs in Holstein cows throughout lactation.

Impact of Climate Change and Heat Stress on Milk Production in Korean Holstein Cows: A Large-Scale Data Analysis.

This study investigated the effects of heat stress on milk production in Korean Holstein cows using large-scale data. Heat stress was assessed using the temperature-humidity index (THI). Weather records (2016 to 2020) were collected from 70 regional weather stations using an installed automated surface observing system (ASOS). A dataset of 2,094,436 milk production records from 215,276 Holstein cows obtained from the Dairy Cattle Genetic Improvement Center was analyzed. Stepwise selection was used to select the input variables, including the daily maximum THI (THI_max). Least-squares means were calculated for milk yield, fat and protein corrected milk (FPCM), fat and protein yield, fat-to-protein ratio, solids not fat, and lactation persistency. Segmented linear regression analysis determined the break points (BPs) of the THI_max. Over the five years, heat stress exposure increased, particularly from May to September. This study identified BPs around THI_max of 80-82 for milk yield and FPCM. Similar patterns for other milk traits were observed, which significantly decreased beyond their respective BPs. These findings indicate that THI variations adversely affect milk yield and composition in dairy cows, highlighting the importance of appropriate feeding management strategies to ensure the optimal productivity of Holstein cows under varying climatic conditions.

A Study on Differential Biomarkers in the Milk of Holstein Cows with Different Somatic Cells Count Levels.

Dairy cow mastitis is one of the common diseases of dairy cows, which will not only endanger the health of dairy cows but also affect the quality of milk. Dairy cow mastitis is an inflammatory reaction caused by pathogenic microorganisms and physical and chemical factors in dairy cow mammary glands. The number of SCC in the milk of dairy cows with different degrees of mastitis will increase in varying degrees. The rapid diagnosis of dairy cow mastitis is of great significance for dairy cow health and farm economy. Based on the results of many studies on the relationship between mastitis and somatic cell count in dairy cows, microflora, and metabolites in the milk of Holstein cows with low somatic cell level (SCC less than 200,000), medium somatic cell level (SCC up to 200,000 but less than 500,000) and high somatic cell level (SCC up to 5000,00) were analyzed by microbiome and metabolic group techniques. The results showed that there were significant differences in milk microbiota and metabolites among the three groups (p < 0.05), and there was a significant correlation between microbiota and metabolites. Meanwhile, in this experiment, 75 differential metabolites were identified in the H group and L group, 40 differential metabolites were identified in the M group and L group, and six differential microorganisms with LDA scores more than four were found in the H group and L group. These differential metabolites and differential microorganisms may become new biomarkers for the diagnosis, prevention, and treatment of cow mastitis in the future.

Genome-Wide Association Study of Lactation Traits in Chinese Holstein Cows in Southern China.

Lactation traits are economically important for dairy cows. Southern China has a high-temperature and high-humidity climate, and environmental and genetic interactions greatly impact dairy cattle performance. The aim of this study was to identify novel single-nucleotide polymorphism sites and novel candidate genes associated with lactation traits in Chinese Holstein cows under high-temperature and humidity conditions in southern China. A genome-wide association study was performed for the lactation traits of 392 Chinese Holstein cows, using GGP Bovine 100 K SNP gene chips. Some 23 single nucleotide polymorphic loci significantly associated with lactation traits were screened. Among them, 16 were associated with milk fat rate, 7 with milk protein rate, and 3 with heat stress. A quantitative trait locus that significantly affects milk fat percentage in Chinese Holstein cows was identified within a window of approximately 0.5 Mb in the region of 0.4-0.9 Mb on Bos taurus autosome 14. According to Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses, ten genes (DGAT1, IDH2, CYP11B1, GFUS, CYC1, GPT, PYCR3, OPLAH, ALDH1A3, and NAPRT) associated with lactation fat percentage, milk yield, antioxidant activity, stress resistance, and inflammation and immune response were identified as key candidates for lactation traits. The results of this study will help in the development of an effective selection and breeding program for Chinese Holstein cows in high-temperature and humidity regions.

Molecular regulatory mechanism of key LncRNAs in subclinical mastitic cows with folic acid supplementation.

BACKGROUND: Folic acid is a water-soluble B vitamin (B9), which is closely related to the body's immune and other metabolic pathways. The folic acid synthesized by rumen microbes has been unable to meet the needs of high-yielding dairy cows. The incidence rate of subclinical mastitis in dairy herds worldwide ranged between 25%~65% with no obvious symptoms, but it significantly causes a decrease in lactation and milk quality. Therefore, this study aims at exploring the effects of folic acid supplementation on the expression profile of lncRNAs, exploring the molecular mechanism by which lncRNAs regulate immunity in subclinical mastitic dairy cows. RESULTS: The analysis identified a total of 4384 lncRNA transcripts. Subsequently, differentially expressed lncRNAs in the comparison of two groups (SF vs. SC, HF vs. HC) were identified to be 84 and 55 respectively. Furthermore, the weighted gene co-expression network analysis (WGCNA) and the KEGG enrichment analysis result showed that folic acid supplementation affects inflammation and immune response-related pathways. The two groups have few pathways in common. One important lncRNA MSTRG.11108.1 and its target genes (ICAM1, CCL3, CCL4, etc.) were involved in immune-related pathways. Finally, through integrated analysis of lncRNAs with GWAS data and animal QTL database, we found that differential lncRNA and its target genes could be significantly enriched in SNPs and QTLs related to somatic cell count (SCC) and mastitis, such as MSTRG.11108.1 and its target gene ICAM1, CXCL3, GRO1. CONCLUSIONS: For subclinical mastitic cows, folic acid supplementation can significantly affect the expression of immune-related pathway genes such as ICAM1 by regulating lncRNAs MSTRG.11108.1, thereby affecting related immune phenotypes. Our findings laid a ground foundation for theoretical and practical application for feeding folic acid supplementation in subclinical mastitic cows.

Metagenomics analysis reveals differences in rumen microbiota in cows with low and high milk protein percentage.

Variation exists in milk protein concentration of dairy cows of the same breed that are fed and managed in the same environment, and little information was available on this variation which might be attributed to differences in rumen microbial composition as well as their fermentation metabolites. This study is aimed at investigating the difference in the composition and functions of rumen microbiota as well as fermentation metabolites in Holstein cows with high and low milk protein concentrations. In this study, 20 lactating Holstein cows on the same diet were divided into two groups (10 cows each), high degree of milk protein group (HD), and low degree of milk protein (LD) concentrations based on previous milk composition history. Rumen content samples were obtained to explore the rumen fermentation parameters and rumen microbial composition. Shotgun metagenomics sequencing was employed to investigate the rumen microbial composition and sequences were assembled via the metagenomics binning technique. Metagenomics revealed that 6 Archaea genera, 5 Bacteria genera, 7 Eukaryota genera, and 7 virus genera differed significantly between the HD and LD group. The analysis of metagenome-assembled genomes (MAGs) showed that 2 genera (g__Eubacterium_H and g__Dialister) were significantly enriched (P < 0.05, linear discriminant analysis (LDA) > 2) in the HD group. However, the LD group recorded an increased abundance (P < 0.05, LDA > 2) of 8 genera (g__CAG-603, g__UBA2922, g__Ga6A1, g__RUG13091, g__Bradyrhizobium, g__Sediminibacterium, g__UBA6382, and g__Succinivibrio) when compared to the HD group. Furthermore, investigation of the KEGG genes revealed an upregulation in a higher number of genes associated with nitrogen metabolism and lysine biosynthesis pathways in the HD group as compared to the LD group. Therefore, the high milk protein concentration in the HD group could be explained by an increased ammonia synthesis by ruminal microbes which were converted to microbial amino acids and microbial protein (MCP) in presence of an increased energy source made possible by higher activities of carbohydrate-active enzymes (CAZymes). This MCP gets absorbed in the small intestine as amino acids and might be utilized for the synthesis of milk protein. KEY POINTS: • Rumen microbiota and their functions differed between cows with high milk protein % and those with low milk protein %. • The rumen microbiome of cows with high milk protein recorded a higher number of enriched genes linked to the nitrogen metabolism pathway and lysine biosynthesis pathway. • The activities of carbohydrate-active enzymes were found to be higher in the rumen of cows with high milk protein %.

Genetic Markers Associated with Milk Production and Thermotolerance in Holstein Dairy Cows Managed in a Heat-Stressed Environment.

Dairy production in Holstein cows in a semiarid environment is challenging due to heat stress. Under such conditions, genetic selection for heat tolerance appears to be a useful strategy. The objective was to validate molecular markers associated with milk production and thermotolerance traits in Holstein cows managed in a hot and humid environment. Lactating cows (n = 300) exposed to a heat stress environment were genotyped using a medium-density array including 53,218 SNPs. A genome-wide association study (GWAS) detected six SNPs associated with total milk yield (MY305) that surpassed multiple testing (p < 1.14 × 10-6). These SNPs were further validated in 216 Holstein cows from two independent populations that were genotyped using the TaqMan bi-allelic discrimination method and qPCR. In these cows, only the SNPs rs8193046, rs43410971, and rs382039214, within the genes TLR4, GRM8, and SMAD3, respectively, were associated (p < 0.05) with MY305, rectal temperature (RT), and respiratory rate. Interestingly, these variables improved as the number of favorable genotypes of the SNPs increased from 0 to 3. In addition, a regression analysis detected RT as a significant predictor (R2 = 0.362) for MY305 in cows with >1 favorable genotype, suggesting this close relationship was influenced by genetic markers. In conclusion, SNPs in the genes TLR4, GRM8, and SMAD3 appear to be involved in the molecular mechanism that regulates milk production in cows under heat-stressed conditions. These SNPs are proposed as thermotolerance genetic markers for a selection program to improve the milk performance of lactating Holstein cows managed in a semiarid environment.

Analysis of Calving Ease and Stillbirth and Their Impact on the Length of Functional Productive Life in Slovak Holstein Cattle.

The aim of the study was to determine the frequency of births according to the categories of calving difficulty and stillbirths and to evaluate the effect of these factors on the longevity of cows. Longevity is one of the traits that affect the overall profit in the dairy industry. A Weibull proportional hazard model was used to evaluate the influence of functional traits such as calving ease and stillbirth. Longevity was expressed as the length of a functional, productive life from the first calving to death or censoring, which was corrected for milk yield. The database included 918,568 calvings, where calving without assistance represented 83.34%, calving with the assistance of one person or the use of a slight mechanical pull represented 14.47%, difficult calving with the assistance of several people, the use of mechanical traction or the intervention of a veterinarian represented 2.16%, and cesarean section represented 0.03%. The mortality of calves, stillborn or dead within 48 h of birth, represented 1.07% and 6.59%, respectively. The frequency of alive female calves was higher (46.84%) than male calves (45.50%). Cows with higher lactations had almost half as many stillborn calves as heifers. The most stillborn calves were found in difficult births (59.48%). In easy calving, this proportion was 2.48%. Using survival analysis, we estimated the significant influence of the factors such as parity, milk production, herd size, age at first calving, herd × year × season, sex of calf, calving ease, and stillbirth on the length of the functional, productive life of cows. The risk of early culling of the cows with moderately difficult calving was 1.259 times higher than in the cows with easy calving. Difficult calving and cesarean section shorten the productive life, and the risk of culling reached 1.711 and 1.894, respectively. Cows that gave birth to a dead calf achieved a 2.939 times higher risk of culling compared to cows that gave birth to a live calf. In this study, a higher risk of early culling was found in cows that gave birth to a male calf. Evaluation of the calving ease and stillbirth can be used as indirect indicators at an earlier age of the animal in the selection process for long-lived animals with good productive and reproductive performance.

Bovine anaplasmosis as a risk factor for retained placenta, mastitis, and abomasal displacement in dairy cattle.

This study aimed to evaluate the frequency of IgG antibodies against A. marginale, the occurrence of this bacterium by qPCR, and the effect of bovine anaplasmosis as a risk factor for clinical cases of retained placenta, mastitis, and abomasal displacement in dairy cattle. For that 179 Holstein cows out of three dairy herds, in the municipality of Sertão, Rio Grande do Sul, Brazil. These cows were on farms that were vulnerable to risk factors that are crucial to susceptibility among these animals to this intracellular hemoparasite. The mean seropositivity for A. marginale from the periods evaluated was 54% on farm A, 69.4% on farm B, and 27.3% on farm C. Molecular diagnosis was performed with qPCR and the mean positivity for A. marginale among the cows on farms A, B, and C in December 2017 was 34.6% (67/179). Infected animals showed clinical cases of retained placenta (6.1%), mastitis (6.1%), and abomasal displacement (0.5%). The association between positivity for anaplasmosis and these clinical cases was assessed through the odds ratio. Our results show that females with a positive qPCR assay for A. marginale had 52.48 times increased probability (OR) to develop clinical cases of retained placenta and mastitis (P < 0.001). These clinical cases negatively impact the productivity of positive females. Thus, implementing preventive and prophylactic control measures to ensure the sanitary quality of the herds is needed to avoid losses due to morbidity and mortality and diminish the economic losses suffered by farmers.

Characteristics of Holstein cows predisposed to ketosis during the post-partum transition period.

BACKGROUND: Ketosis is a common metabolic disorder during the post-partum transition period of dairy cattle. How the method of reproduction, parturition time, and calf birth weight affect the occurrence of ketosis on dairy herds remains elusive. OBJECTIVES: This study investigated factors associated with the severity of ketosis. METHODS: We divided 186 Holstein cows into three classifications based on the highest ß-hydroxybutyrate (BHBA) concentration during the post-partum transition period, namely non-ketosis (<1.2 mmol/L, n = 94), subclinical ketosis (1.2-2.9 mmol/L, n = 58), and clinical ketosis (≥3.0 mmol/L, n = 34). We evaluated characteristics of cows associated with the severity of ketosis. RESULTS: Ketosis was not associated with the method of reproduction, parturition time, pregnancy wastage, premature delivery, retained placenta, and type of calf. Cows calving in spring and especially summer were at higher risk of severe ketosis (p < 0.01). Cows with increased body condition score (BCS) at parturition, age, lactation number, and calving interval were more likely to develop severe ketosis (p < 0.05). Cows with clinical ketosis produced most milk (29.9 ± 1.0 kg) from days four to six, whereas cows without ketosis produced the least (21.3 ± 0.8 kg) (p < 0.001). Heavier calf birth weight resulted in high risk of severe ketosis (p < 0.01), due to increased milk yield during the early lactation. CONCLUSIONS: The severity of ketosis is associated with the calving season, BCS at parturition, age, lactation number, calving interval, milk yield in the early lactation period, and calf birth weight. Nonetheless, it was not associated with the method of reproduction, parturition time, pregnancy wastage, premature delivery, retained placenta, and type of calf. This study is the first to investigate the associations between ketosis and calf birth weight. Our findings could help predict cows at risk of ketosis and take precautions.

Fermented soybean meal modified the rumen microbiome to enhance the yield of milk components in Holstein cows.

The study was conducted to evaluate the rumen microbiota as well as the milk composition and milk component yields of Holstein cows supplemented with fermented soybean meal (FSBM). Eighteen Holstein cows in their 2nd parity with 54.38 ± 11.12 SD days in milking (DIM) were divided into two dietary groups (CON and TRT) of nine cows per group. The cows in the TRT group received 300 g of FSBM per cow per day in addition to the conventional diet, while each cow in the CON group was supplemented with 350 g of soybean meal (SBM) in their diet daily throughout the 28-day feeding trial. Rumen bacterial composition was detected via 16S rRNA sequencing, and the functional profiles of bacterial communities were predicted. Milk composition, milk yield, as well as rumen fermentation parameters, and serum biochemistry were also recorded. The inclusion of FSBM into the diets of Holstein cows increased the milk urea nitrogen (MUN), milk protein yield, fat corrected milk (FCM), and milk fat yield while the milk somatic cell count (SCC) was decreased. In the rumen, the relative abundances of Fibrobacterota, and Spirochaetota phyla were increased in the TRT group, while the percentage of Proteobacteria was lower. In addition, the supplementation of FSBM to Holstein cows increased the acetate percentage, rumen pH, and acetate to propionate ratio, while the proportion of propionate and propionate % was observed to decrease in the TRT group. The KEGG pathway and functional prediction revealed an upregulation in the functional genes associated with the biosynthesis of amino acids in the TRT group. This enrichment in functional genes resulted in an improved synthesis of several essential amino acids including lysine, methionine, and branch chain amino acids (BCAA) which might be responsible for the increased milk protein yield. Future studies should employ shotgun metagenomics, transcriptomics, and metabolomics technology to investigate the effects of FSBM on other rumen microbiomes and milk protein synthesis in the mammary gland in Holstein cows. KEY POINTS: • The supplementation of fermented soybean meal (FSBM) to Holstein cows modified the proportion of rumen bacteria. • Predicted metabolic pathways and functional genes of rumen bacteria revealed an enrichment in pathway and genes associated with biosynthesis of amino acids in the group fed FSBM. • The cows supplemented with FSBM record an improved rumen fermentation. • Cows supplemented with FSBM recorded an increased yield of milk protein and milk fat.

Hematology Reference Intervals for Holstein Cows in Southern China: A Study of 786 Subjects.

Hematology RIs help clinicians and researchers determine whether a hematology parameter is abnormal, which plays an important role in animal health surveillance. China is one of the largest dairy producers in the world, with millions of Holstein cows. However, there has been no published data on hematology RIs for dairy cows in China yet. Therefore, the aim of this study is to establish updated and accurate RIs for Holstein cows in southern China. To increase the accuracy of the RIs, we collected blood samples from 786 Holstein cows and analyzed 25 hematologic variables. The RIs for Holstein cows were established using the 95% percentile RIs according to the American Society of Veterinary Clinical Pathology guidelines. The effects of different ages, parities and lactation stages were also checked in this study. The data of 21, 22 and 19 out of 25 hematology parameters were significantly different between different ages, parities and lactation stages, respectively. Furthermore, the hematology RIs of separate subclasses according to different ages, parities and lactation stages were generated. Hematology RIs according to ages and lactation stages, as well as parities and lactation stages, were also assessed. Together, our results confirm that hematology RIs for cows vary by ages, parities and lactation stages. The present study helps to fill the gap in hematology RIs for Holstein cows in southern China, and our data may serve as a very useful tool for monitoring the health and welfare of dairy cattle in China.

Predicting methane emission in Canadian Holstein dairy cattle using milk mid-infrared reflectance spectroscopy and other commonly available predictors via artificial neural networks.

Interest in reducing eructed CH4 is growing, but measuring CH4 emissions is expensive and difficult in large populations. In this study, we investigated the effectiveness of milk mid-infrared spectroscopy (MIRS) data to predict CH4 emission in lactating Canadian Holstein cows. A total of 181 weekly average CH4 records from 158 Canadian cows and 217 records from 44 Danish cows were used. For each milk spectra record, the corresponding weekly average CH4 emission (g/d), test-day milk yield (MY, kg/d), fat yield (FY, g/d), and protein yield (PY, g/d) were available. The weekly average CH4 emission was predicted using various artificial neural networks (ANN), partial least squares regression, and different sets of predictors. The ANN architectures consisted of 3 training algorithms, 1 to 10 neurons with hyperbolic tangent activation function in the hidden layer, and 1 neuron with linear (purine) activation function in the hidden layer. Random cross-validation was used to compared the predictor sets: MY (set 1); FY (set 2); PY (set 3); MY and FY (set 4); MY and PY (set 5); MY, FY, and PY (set 6); MIRS (set 7); and MY, FY, PY, and MIRS (set 8). All predictor sets also included age at calving and days in milk, in addition to country, season of calving, and lactation number as categorical effects. Using only MY (set 1), the predictive accuracy (r) ranged from 0.245 to 0.457 and the root mean square error (RMSE) ranged from 87.28 to 99.39 across all prediction models and validation sets. Replacing MY with FY (set 2; r = 0.288-0.491; RMSE = 85.94-98.04) improved the predictive accuracy, but using PY (set 3; r = 0.260-0.468; RMSE = 86.95-98.47) did not. Adding FY to MY (set 4; r = 0.272-0.469; RMSE = 87.21-100.76) led to a negligible improvement compared with sets 1 and 3, but it slightly decreased accuracy compared with set 2. Adding PY to MY (set 5; r = 0.250-0.451; RMSE = 87.66-100.94) did not improve prediction ability. Combining MY, FY, and PY (set 6; r = 0.252-0.455; RMSE = 87.74-101.93) yielded accuracy slightly lower than sets 2 and 3. Using only MIRS data (set 7; r = 0.586-0.717; RMSE = 69.09-96.20) resulted in superior accuracy compared with all previous sets. Finally, the combination of MIRS data with MY, FY, and PY (set 8; r = 0.590-0.727; RMSE = 68.02-87.78) yielded similar accuracy to set 7. Overall, sets including the MIRS data yielded significantly better predictions than the other sets. To assess the predictive ability in a new unseen herd, a limited block cross-validation was performed using 20 cows in the same Canadian herd, which yielded r = 0.229 and RMSE = 154.44, which were clearly much worse than the average r = 0.704 and RMSE = 70.83 when predictions were made by random cross-validation. These results warrant further investigation when more data become available to allow for a more comprehensive block cross-validation before applying the calibrated models for large-scale prediction of CH4 emissions.