Dataset Information

Proteome response to microbiota in dairy cows with high and low somatic cell count

ABSTRACT: Host response to invasive microbes in the bovine udder has an important role on the animal health and is essential to the dairy industry to ensure production of high-quality milk and reduce the mastitis incidence. To better understand the biology behind these host-microbiome interactions, we investigated the somatic cell proteomes at quarter level for four cows (collected before and after milking) using a shotgun proteomics approach. Simultaneously, we identified the quarter microbiota by amplicon sequencing to detect presence of mastitis pathogens or other commensal taxa. In total, 32 quarter milk samples were analyzed divided in two groups depending on the somatic cell count (SCC). The high SCC group (> 100,000 cell/mL) included 10 samples and significant different proteome profiles were detected. Differential abundance analysis uncovers a specific expression pattern in high SCC samples revealing pathways involved in immune responses such as inflammation, activation of the complement system, migration of immune cells, and tight junctions.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Bos Taurus (bovine)

SUBMITTER: Davide Porcellato

LAB HEAD: Davide Porcellato

PROVIDER: PXD035328 | Pride | 2023-05-10

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20211025_JT10_Slot2-10_1_887.d.zip	Other
	20211025_JT11_Slot2-11_1_888.d.zip	Other
	20211025_JT12_Slot2-12_1_889.d.zip	Other
	20211025_JT13_Slot2-13_1_891.d.zip	Other
	20211025_JT14_Slot2-14_1_892.d.zip	Other

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Publications

Metataxonomic analysis and host proteome response in dairy cows with high and low somatic cell count: a quarter level investigation.

Winther Anja Ruud AR da Silva Duarte Vinícius V Porcellato Davide D

Veterinary research 20230404 1

Host response to invasive microbes in the bovine udder has an important role on the animal health and is essential to the dairy industry to ensure production of high-quality milk and reduce the mastitis incidence. To better understand the biology behind these host-microbiome interactions, we investigated the somatic cell proteomes at quarter level for four cows (collected before and after milking) using a shotgun proteomics approach. Simultaneously, we identified the quarter microbiota by amplic ...[more]

PMID: 37016420

Similar Datasets

Project description:Bovine mastitis, the infection of the mammary gland which leads to great health and economic challenges for dairy farmers is accompanied by dramatic changes in the milk proteome. In this study of naturally occurring mastitis not only have the changes in the milk proteome been quantified in subclinical and clinical mastitis but simultaneous changes in the serum proteome have also been characterised and quantified. Milk and serum samples from healthy dairy cows (n=12) were compared to those of cows with subclinical (n=10) and clinical mastitis (n=112) using TMT label-based proteomic approach. The study included the milk and serum samples taken from thirty-two dairy cows ( kept on private farms located in Croatia. All cows were checked by physical examination. Somatic cells count (SCC) and mastitis test in milk samples were performed. According to the results, cows were assigned into three groups: Group I (control, n=10) consisted of healthy cows with SCC below 400,000 cells/ml on the monthly check-up and a negative mastitis test and without any clinical sign of mastitis. Group II (subclinical mastitis, n=12) comprised cows without clinical signs of mastitis but with SCC above 400,000 cells/ml on the monthly basis and a positive mastitis test at the time of sampling. Group III (clinical mastitis, n=10) consisted of cows with clinical signs of mastitis which include changes in milk appearance (flakes and clots in milk), different stages of udder inflammation (hyperemia, edema, pain, udder enlargement and elevated udder temperature) and disturbance of general health (depression, relaxed cold ears, dehydration, elevated body temperature, increased heart and respiratory rate, decreased ruminal contraction and decreased appetite). Blood samples were taken from v. coccygea and centrifuged at 3000 g for 15 min after clotting for two hours at room temperature. Serum samples were stored at -80°C until analysis. Milk samples were taken aseptically before the morning milking. First few streams were discarded. Teat ends were disinfected with cotton swabs soaked with 70% ethanol. Samples were taken into sterile tubes and transported to laboratory on ice within a few hours.

Project description:In a recent study, a teat disinfectant based on a nisin-producing Lactococcus cremoris displayed comparable efficacy to a conventional iodophor disinfectant in preventing dairy cow mastitis when implemented in the dairy cow milking routine. Here, we carried out a longitudinal study for an in-depth investigation of the milk proteome. Four groups of matched quarter milk with low and high somatic cell count (SCC) collected at the beginning (T0) and at the end (TF) of the trial were selected for analysis, for a total of 28 low SCC (LSCC) and 12 high SCC (HSCC) samples. The milk was delipidated, digested into peptides, and subjected to a proteomic analysis workflow entailing NanoHPLC separation and tandem mass spectrometry (LC-MS/MS) on an Orbitrap Fusion Tribrid mass spectrometer. The proteins were identified with MaxQuant, the proteomes were compared for identifying differential proteins, and their interaction networks were assessed with STRING. In LSCC milk at T0, 31 were increased or present only in the Lactococcus (L) disinfectant group (from now INCREASED in L) and 59 proteins were decreased in L disinfectant group or present only in the conventional iodophor (C) group (from now DECREASED in L). All were involved in metabolic pathways and were likely related to physiological differences between animals. In LSCC milk at TF, 34 proteins were INCREASED in L and 46 proteins were DECREASED in the L disinfectant group. Most belonged to physiological milk pathways, but 3 antimicrobial proteins were enriched in the L group at the end of the trial. In HSCC milk at T0, 54 proteins were INCREASED in L and 34 DECREASED in L. Most were involved in physiological metabolic pathways; 4 proteins with antimicrobial functions were higher in L at the beginning of the trial. In HSCC milk at TF, 67 proteins were INCREASED in L and 39 DECREASED in L. All the proteins enriched in C belonged to metabolic pathways, while up to 14 proteins enriched in L were involved in antibiotic, antimicrobial, and immune defense pathways. Cathelicidins were among the most prominent differential proteins with antimicrobial functions. At T0, cathelicidins were generally higher in HSCC than LSCC milk of both experimental groups, as expected. At TF, they were increased or unique only in the Lactococcus group, in both LSCC and HSCC milk. Western immunoblotting validation confirmed the proteomic findings. On the other hand, enzymatic assessment of n-acetyl-glucosaminidase (NAGase) did not highlight significant differences between the C and L groups, neither for LSCC nor for HSCC milk. In conclusion, the two disinfectants did not induce relevant functional changes in the milk proteome of healthy animals, except for three increased antimicrobial proteins in the Lactococcus group. On the other hand, in animals with an underlying mammary gland inflammation, the milk proteome of the Lactococcus group showed increased levels of several proteins with antibiotic, antimicrobial and immune defense functions. Among other factors, the selective immunostimulatory action of nisin, the antimicrobial effector of L. cremoris, might play a role in this observation.