Search

Simple Summary: Milk fat is one of the most important economic traits in dairy cow production. Numerous investigations have demonstrated the influence of the ruminal microbiome's composition and metabolic profile on milk production. However, no studies have directly connected the ruminal metabolome with bacterial and fungal communities as a function of milk fat content in dairy cows. The specific objective of the present study was to compare ruminal metabolome with bacterial and fungal communities among Chinese Holstein cows with contrasting milk fat content that were fed the same diet. Our results confirm that ruminal metabolome, bacterial, and fungal communities differ significantly in dairy cows with different milk fat contents. Among these different metabolites, lauric acid was enriched in fatty acid biosynthesis and selected for an in vitro study with bovine mammary epithelial cells, indicating that lauric acid promoted intracellular triglyceride synthesis via upregulation of the mRNA abundance of fatty acid uptake and activation, as well as lipogenesis. This study revealed microbial-mediated metabolic processes related to milk fat synthesis. The composition and metabolic profile of the ruminal microbiome have an impact on milk composition. To unravel the ruminal microbiome and metabolome affecting milk fat synthesis in dairy cows, 16S rRNA and internal transcribed spacer (ITS) gene sequencing, as well as ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS/MS) methods were used to investigate the significant differences in ruminal bacterial and fungal communities as well as metabolome among Chinese Holstein cows with contrasting milk fat contents under the same diet (H-MF 5.82 ± 0.41% vs. L-MF 3.60 ± 0.12%). Another objective was to culture bovine mammary epithelial cells (BMECs) to assess the effect of metabolites on lipid metabolism. Results showed that the acetate-to-propionate ratio and xylanase activity in ruminal fluid were both higher in H-MF. Microbiome sequencing identified 10 types of bacteria and four types of fungi differently abundant at the genus level. Metabolomics analysis indicated 11 different ruminal metabolites between the two groups, the majority of which were lipids and organic acids. Among these, lauric acid (LA) was enriched in fatty acid biosynthesis with its concentration in milk fat of H-MF cows being greater (217 vs. 156 mg per 100 g milk), thus, it was selected for an in vitro study with BMECs. Exogenous LA led to a marked increase in intracellular triglyceride (TG) content and lipid droplet formation, and it upregulated the mRNA abundance of fatty acid uptake and activation (CD36 and ACSL1), TG synthesis (DGAT1, DGAT2 and GPAM), and transcriptional regulation (SREBP1) genes. Taken together, the greater relative abundance of xylan-fermenting bacteria and fungi, and lower abundance of bacteria suppressing short-chain fatty acid-producing bacteria or participating in fatty acid hydrogenation altered lipids and organic acids in the rumen of dairy cows. In BMECs, LA altered the expression of genes involved in lipid metabolism in mammary cells, ultimately promoting milk fat synthesis. Thus, it appears that this fatty acid plays a key role in milk fat synthesis. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The cow's milk production characteristics are a significant economic indicator in the livestock industry. Serum cytokines such as interleukin-17 (IL-17) may be potential indicators for bovine mastitis concerning the milk somatic cell count (SCC) and somatic cell score (SCS). The current study aims to find previously undiscovered single nucleotide polymorphisms in the bovine (IL-17A) gene and further investigates their associations with milk production traits in Chinese Holstein cows. Twenty Chinese Holstein cows were randomly chosen from six farms in Jiangsu Province, China. The DNA was extracted from selected samples of bloods for PCR amplification Sequence analyses were used to find SNPs in the bovine (IL-17A) gene. The discovered five SNPs are g-1578A>G, g-1835G>A, and g-398T>A in the 5′UTR; g3164T>C and g3409G>C in the exon region. The genotyping of Holstein cows (n = 992) was performed based on Sequenom Mass ARRAY and SNP data. The connection between SNPs, milk production variables, and the somatic cell score was investigated using the least-squares method. Based on the results, SNP g-398T>A had a significant linkage disequilibrium with g3164T>C. SNPs were found to have significant (p < 0.05) correlations with the test-day milk yield. In conclusion, IL-17A affects cow's milk production traits significantly. [ABSTRACT FROM AUTHOR]

This research paper aimed to explore the characteristics of Holstein cattle's milk fat percentage lactation curve and its influencing factors. The Wood model was used for fitting the lactation curve of 398,449 DHI test-day milk fat percentage records of Holstein cows from 2018 to 2020 in 12 dairy farms in Jiangsu province, and the influencing factors—including farm size, parity, calving season, calving interval, and 305-days milk production—on the parameters of the lactation curve were analyzed. The results showed that the non-genetic factors such as dairy farm size, calving season, parity, calving interval, and 305-days milk yield have a significant impact on milk fat percentage (p < 0.01); the average R2 of the daily milk fat percentage curve was 0.9699; the lowest milk fat percentage was 3.54%; the time to reach the lowest milk fat percentage was 126 days; and the persistence of milk fat percentage was 3.59%. All of these factors explored in this study fit at different levels above 0.96. The Wood model performed well in the fitting and analysis of the milk fat percentage curve of Holstein cattle in Jiangsu Province. This study provides a reference for improving the milk fat percentage of Holstein cattle. [ABSTRACT FROM AUTHOR]

In this study, circular RNAs (circRNAs) from Holstein cow mammary tissues were identified and compared between early lactation and non-lactation. After analysis, 10,684 circRNAs were identified, ranging from 48 to 99,406 bp, and the average size was 882 bp. The circRNAs were mainly distributed on chromosomes 1 to 11, and 89.89% of the circRNAs belonged to sense-overlapping circRNA. The exons contained with circRNAs ranged from 1 to 47 and were concentrated from 1 to 5. Compared with the non-lactating cows, 87 circRNAs were significantly differentially expressed in the peak lactation cows. There were 68 upregulated circRNAs and 19 downregulated circRNAs. Enrichment analysis of circRNAs showed that GO analysis mainly focused on immune response, triglyceride transport, T cell receptor signaling pathway, etc. Pathway analysis mainly focused on cytokine-cytokine receptor interaction, T helper 17 cell differentiation, fatty acid biosynthesis, the JAK-STAT signaling pathway, etc. Specific primers were designed for two proximal ends of the circRNA junction sites to allow for PCR validation of four randomly selected circRNAs and carry out circRNA-miRNA interaction research. This study revealed the expression profile and characteristics of circRNAs in mammary tissue from Holstein cows at early lactation and non-lactation, thus providing rich information for the study of circRNA functions and mechanisms, as well as potential candidate miRNA genes for studying lactation in Holstein cows. [ABSTRACT FROM AUTHOR]

The current study reports the identification of previously undiscovered single-nucleotide polymorphisms (SNPs) in the bovine AGPAT3 gene and further investigates their associations with milk production traits. Our results demonstrate that the major allele C of the SNP g.12264 C > T is positively correlated with test-day milk yield, protein percentage and 305-day milk yield. Importantly, in silico analysis showed that the C/T transition at this locus gives rise to two new transcription factor binding sites (TFBS), E2F1 and Nkx3-2. Polymorphism g.18658 G > A was the only SNP associated with milk urea nitrogen (MUN) with the G allele related to an increase in milk urea nitrogen as well as fat percentage. The GG genotype of SNP g.28731 A > G was associated with the highest fat and protein percentage and lowest 305-day milk yield and somatic cell score (SCS). The association between AGPAT3 locus and milk production traits could be utilized in marker-assisted selection for the genetic improvement of milk production traits and, probably in conjunction with other traits, for selection to improve fitness of dairy cattle. [ABSTRACT FROM AUTHOR]

We established a mastitis model using exogenous infection of the mammary gland of Chinese Holstein cows with Staphylococcus aureus and extracted total RNA from S. aureus -infected and healthy mammary quarters. Differential expression of genes due to mastitis was evaluated using Affymetrix technology and results revealed a total of 1230 differentially expressed mRNAs. A subset of affected genes was verified via Q-PCR and pathway analysis. In addition, Solexa high-throughput sequencing technology was used to analyze profiles of miRNA in infected and healthy quarters. These analyses revealed a total of 52 differentially expressed miRNAs. A subset of those results was verified via Q-PCR. Bioinformatics techniques were used to predict and analyze the correlations among differentially expressed miRNA and mRNA. Results revealed a total of 329 pairs of negatively associated miRNA/mRNA, with 31 upregulated pairs of mRNA and 298 downregulated pairs of mRNA. Differential expression of miR-15a and interleukin-1 receptor-associated kinase-like 2 (IRAK2), were evaluated by western blot and luciferase reporter assays. We conclude that miR-15a and miR-15a target genes (IRAK2) constitute potential miRNA–mRNA regulatory pairs for use as biomarkers to predict a mastitis response. [ABSTRACT FROM AUTHOR]

Dairy cow mastitis is a detrimental factor in milk quality and food safety. Mastitis generally refers to inflammation caused by infection by pathogenic microorganisms. Our studies in recent years have revealed the role of miRNA regulation in Staphylococcus aureus‐induced mastitis. In the present study, we overexpressed and suppressed miR‐145 to investigate the function of miR‐145 in Mac‐T cells. Flow cytometry, ELISA and EdU staining were used to detect changes in the secretion of several Mac‐T cytokines and in cell proliferation. We found that overexpression of miR‐145 in Mac‐T cells significantly reduced the secretion of IL‐12 and TNF‐α, but increased the secretion of IFN‐γ; the proliferation of bovine mammary epithelial cells was also inhibited. Using quantitative real‐time PCR (qRT‐PCR), Western blotting and luciferase multiplex verification techniques, we found that miR‐145 targeted and regulated FSCN1. Knock‐down of FSCN1 significantly increased the secretion of IL‐12, while the secretion of TNF‐α was significantly downregulated in Mac‐T cells. Upon S. aureus infection of mammary gland tissue, the body initiated inflammatory responses; Bta‐miR‐145 expression was downregulated, which reduced the inhibitory effect on the FSCN1 gene; and upregulation of FSCN1 expression promoted mammary epithelial cell proliferation to allow the recovery of damaged tissue. The results of the present study will aid in understanding the immune mechanism opposing S. aureus infection in dairy cows and will provide a laboratory research basis for the prevention and treatment of mastitis. [ABSTRACT FROM AUTHOR]

The research reported in this Research Communication aimed to describe the influence of Toll-like receptor 4 gene polymorphisms on milk production traits in Chinese Holstein cows. Toll-like receptor 4 (TLR4) is an important member of the toll-like receptor gene family that is widely found in various organisms. Since TLR4 can identify molecular patterns from various pathogenic microorganisms and induce natural and acquired immunity, it plays an important role in disease resistance in dairy cows. Two single nucleotide polymorphisms (SNPs) of TLR4 (c.−226 G > C and c.2021 C > T) that were previously found to be associated with health traits were genotyped using Sequenom MassARRAY (Sequenom Inc., San Diego, CA) for Chinese Holstein cows (n = 866). The associations between SNPs or their haplotypes and milk production traits and somatic cell count were analyzed by the generalized linear model procedure of Statistics Analysis System software (SAS). The c.−226 G > C and c.2021 C > T showed low linkage disequilibrium (r 2 = 0·192). There was no association between these two SNPs and SCC, but significant effects were found for SNP c.−226 G > C on test-day milk yield, fat content, protein content, and total solid and milk urea nitrogen (P < 0·05), and SNP c.2021 C > T and the SNP haplotypes on test-day milk yield, fat content, protein content, lactose content and total solids (P < 0·05). The software MatInspector revealed that c.−226 G > C was located within several potential transcription factor binding sites, including transcription factor AP-2. The polymorphisms c.−226 G > C and c.2021 C > T had significant effects on the milk production for Chinese Holstein, and these SNP could be used for molecular marker-assisted selection of milk production. [ABSTRACT FROM AUTHOR]

Simple Summary: Milk production traits of cows are important economic indicators of the livestock industry. Many dairy farms strive to improve the quality of their milk. Long-chain acyl-CoA synthetase 1 (ACSL1) is a gene related to lipid metabolism. It is widely found in various organisms and can affect fat content and protein content in milk. Single nucleotide polymorphisms (SNP) refers to the polymorphism of DNA sequence caused by a single nucleotide variation at the gene level, which plays a vital function in the genetic study of milk production traits in dairy cows. Our study identified six SNPs of the ACSL1 gene in Chinese Holstein cows, which were related to milk yield, milk fat content, milk protein content and somatic cell score (SCS) to some extent. In summary, the pleiotropic effects of bovine ACSL1 for milk production traits were found in this paper, which will provide a reference for Chinese Holstein cow breeding selection and high economic benefits. Improving the quality of milk is a challenge for zootechnicians and dairy farms across the globe. Long-chain acyl-CoA synthetase 1 (ACSL1) is a significant member of the long-chain acyl-CoA synthetase gene family. It is widely found in various organisms and influences the lactation performance of cows, including fat percentage, milk protein percentage etc. Our study was aimed to investigate the genetic effects of single nucleotide polymorphisms (SNPs) in ACSL1 on milk production traits. Twenty Chinese Holstein cows were randomly selected to extract DNA from their blood samples for PCR amplification and sequencing to identify SNPs of the bovine ACSL1 gene, and six SNPs (5'UTR-g.20523C>G, g.35446C>T, g.35651G>A, g.35827C>T, g.35941G>A and g.51472C>T) were discovered. Then, Holstein cow genotyping (n = 992) was performed by Sequenom MassARRAY based on former SNP information. Associations between SNPs and milk production traits and somatic cell score (SCS) were analyzed by the least-squares method. The results showed that SNP g.35827C>T was in high linkage disequilibrium with g.35941G>A. Significant associations were found between SNPs and test-day milk yield (TDMY), fat content (FC), protein content (PC) and SCS (p < 0.05). Among these SNPs, SNP 5'UTR-g.20523C>G showed an extremely significant effect on PC and SCS (p < 0.01). The SNP g.35446C>T showed a statistically significant effect on FC, PC, and SCS (p < 0.01), and also TDMY (p < 0.05). The SNP g.35651G>A had a statistically significant effect on PC (p < 0.01). The SNP g.35827C>T showed a highly significant effect on TDMY, FC, and SCS (p < 0.01) and significantly influenced PC (p < 0.05). Lastly, SNP g.51472C>T was significantly associated with TDMY, FC, and SCS (p < 0.05). In summary, the pleiotropic effects of bovine ACSL1 for milk production traits were found in this paper, but further investigation will be required on the intrinsic correlation to provide a theoretical basis for the research on molecular genetics of milk quality traits of Holstein cows. [ABSTRACT FROM AUTHOR]

Simple Summary: Searching for causative polymorphisms underlying the variability of milk production traits and then incorporating them into breeding programs are very effective ways to improve the efficiency and reliability of conventional dairy cattle breeding. Fatty acid desaturase 2 (FADS2) plays a pivotal role in the biosynthesis of polyunsaturated fatty acids. Previous studies provided evidence that FADS2 was one of the most downregulated genes during negative energy balance in the liver of postpartum dairy cattle. Genes involved in the energetic pathways may influence other production traits, such as protein, fat and milk yields. Therefore, in the present study, we investigated the common genetic variants of the FADS2 gene in Chinese Holstein cows. Our results provided direct evidence that FADS2 was an interesting candidate for selection to increase milk production and improve resistance against mastitis. This study investigated the single nucleotide polymorphisms (SNPs) of Fatty acid desaturase 2 (FADS2) gene and further explored their genetic effects on conventionally collected milk production traits in Chinese Holstein cows using 18,264 test-day records of 841 cows. One missense mutation c. 908 C > T (SNP site in the complementary DNA sequence), which caused an amino acid change from alanine to valine (294Ala > Val), and two 3' untranslated region (UTR) SNPs, c.1571 G > A and c.2776 A > G were finally identified. The SNP c.908 C > T was significantly associated with test-day milk yield, fat percentage and 305-day milk, fat and protein yield. In particular, the T allele of the SNP c.908 C > T showed a significant association with decreased somatic cell score (SCS) in the investigated population. Significant relationship between the SNP c.1571 G > A and 305-day milk yield showed that genotype GG was linked to the highest milk yield. Substituting the allele G for A at the c.2776 A > G locus resulted in a decrease of protein percentage. Our results demonstrated that FADS2 was an interesting candidate for selection to increase milk production and improve resistance against mastitis. [ABSTRACT FROM AUTHOR]