Project description:The proteins of the milk fat globule membrane (MFGM) have a number of functions, such as the regulation of milk fat secretion and metabolism, the uptake and transportation of fatty acids in the intestine and potential protection from bacterial or viral infection. While the proteome of the MFGM in bovine milk has been extensively characterised, our knowledge of these proteins in buffalo milk is limited. In this study, a proteomic approach was used to characterise the proteome of the buffalo MFGM. Multiple extraction techniques were employed to increase the coverage of proteins identified, while label free relative quantitative liquid chromatography tandem mass spectrometry was used for comparison between the buffalo and bovine MFGM proteome. A total of 220 buffalo MFGM proteins and 234 bovine MFGM proteins were identified after being filtered from the initial dataset of 757 and 680 proteins respectively. Significantly greater amounts of xanthine oxidoreductase, platelet glycoprotein 4, heat shock cognate and calcineurin B homologous protein were identified per mass of buffalo MFGM protein extracted. The higher expression of xanthine oxidase in the MFGM of buffalo milk was confirmed by Western blot analysis and a heterogeneous distribution of this protein observed in situ on the surface of the MFGM. The high concentration of fat in buffalo milk, together with the differences in the MFGM proteome indicate differences in the nutritional profile, biological function, and potential ease of processing of buffalo and bovine milk products.
Project description:Infectious mastitis by non-aureus staphylococci (NAS) is a significant issue in dairy buffalo farming. In a herd with subclinical NAS mastitis, we identified Staphylococcus microti as the predominant species by MALDI-TOF mass spectrometry. To assess milk protein integrity and investigate potential disease markers, we characterized 12 NAS-positive and 12 healthy quarter milk samples by shotgun peptidomics combining peptide enrichment and high-performance liquid chromatography/tandem mass spectrometry (LC-MS/MS). We observed significant changes in the milk peptidome. Out of 789 total peptides identified in each group, 49 and 44 were unique or increased in NAS-positive and healthy milk, respectively. In terms of sequence, these belonged mainly to caseins in NAS-positive milk, followed by milk fat globule membrane proteins (MFGMP) and by the immune defense/antimicrobial proteins osteopontin, lactoperoxidase, and serum amyloid A. In healthy milk, the differential peptides belonged mainly to MFGMP, followed by caseins. In terms of abundance, peptides from MFGMP and immune defense protein were higher in NAS-positive milk, while peptides from caseins were higher in healthy milk. These findings highlight the impact of NAS on buffalo milk quality and mammary gland health, even when clinical signs are not evident, and underscore the need for clarifying the epidemiology and relevance of the different NAS species in this dairy ruminant.
2022-06-09 | PXD028793 | Pride
Project description:Polymorphic SSR identification in Eleutheronema tetradactylum (Indian Salmon)
Project description:The 4 day gene expression profile in the lactating mammary gland was demonstrated using the Human Ref-8 BeadChip array (Illumina, Inc). Of the 22,184 gene transcripts on the array, 14,070 genes were consistently expressed and represented the milk fat globule transcriptome. Milk protein genes were among the most highly expressed along with genes involved in the milk syntesis processes. Serial milk samples were collected every 3h for 4 days from 5 normal, lactating women. RNA was isolated from the milk fat globules and utilized for microarray analyses.
Project description:We have utilized the RNA isolated from breast milk fat globule (MFG) from lactating women from 6h to 42 days following delivery using the HumanHT-12 v4 Expression BeadChip (Illumina, Inc) to determine the temporal coordination of changes in gene expression in milk substrate synthesis processes. Of the 47,323 gene transcripts on the array, 16,623 transcripts were expressed. Major milk protein genes were among the most highly expressed and induced along with those involved in the metabolic and biosynthetic processes of carbohydrate and lipid. Milk fat synthesis mirrors expression of genes involved in fat synthesis, lipolysis and transport. In the lactose synthesis pathway, expression of α-lactalbumin mRNA was high already in the early milk and did not change. Lactose synthesis paralleled the induction of gene expression of proteins involved in UDP-galactose synthesis and transport. Serial milk samples were collected 6h following delivery, q12 h for the first 4 days and then weekly for 6 wks from 7 healthy, lean, exclusively breastfeeding women [7 x 15 = 105 samples]. RNA was isolated from the milk fat globules and utilized for microarray analyses.
Project description:Background: Intramuscular fat (IMF) content is an important index for beef quality. However, the genetics of IMF deposition is complex and still largely unclear, especially in buffalo. To identify miRNAs with potential regulatory role in lipid accumulated in muscle, we performed small RNA sequencing and identified miRNAs expressed in the longissimus dorsi muscle and back fat of Chinese buffalo, which provided vital information for further identification of miRNAs with potential regulatory role in the lipid accumulated in muscle. Results: Three small RNA libraries were constructed. A total of 32762032 raw reads were obtained from adipose groups, respectively. After filtering the adaptor and low quality reads, 32054381 clean reads were retained. In total, 623 miRNAs were identified.
Project description:The 4 day gene expression profile in the lactating mammary gland was demonstrated using the Human Ref-8 BeadChip array (Illumina, Inc). Of the 22,184 gene transcripts on the array, 14,070 genes were consistently expressed and represented the milk fat globule transcriptome. Milk protein genes were among the most highly expressed along with genes involved in the milk syntesis processes.
Project description:The 24h gene expression profile in the lactating mammary gland was demonstrated using the Human Ref-8 BeadChip array (Illumina, Inc). Of the 22,184 gene transcripts on the array, 14,070 genes were consistently expressed and represented the milk fat globule transcriptome. Milk protein genes were among the most highly expressed along with genes involved in the milk syntesis processes. Keywords: time course analysis
Project description:Background: Intramuscular fat (IMF) content is an important index for beef quality. However, the genetics of IMF deposition is complex and still largely unclear, especially in buffalo. To identify miRNAs with potential regulatory role in lipid accumulated in muscle, we performed small RNA sequencing and identified miRNAs expressed in the longissimus dorsi muscle and back fat of Chinese buffalo, which provided vital information for further identification of miRNAs with potential regulatory role in the lipid accumulated in muscle. Results: Six small RNA libraries were constructed. A total of 66,128,645 and 70,974,347 raw reads were obtained from muscle and adipose groups, respectively. After filtering the adaptor and low quality reads, 60,765,257 and 67,327,095 clean reads were retained. In total, 721 miRNAs were identified.