Rumen-protected methionine compared with rumen-protected choline improves immunometabolic status in dairy cows during the peripartal period.

The immunometabolic status of peripartal cows is altered due to changes in liver function, inflammation, and oxidative stress. Nutritional management during this physiological state can affect the biological components of immunometabolism. The objectives of this study were to measure concentrations of biomarkers in plasma, liver tissue, and milk, and also polymorphonuclear leukocyte function to assess the immunometabolic status of cows supplemented with rumen-protected methionine (Met) or choline (CHOL). Forty-eight multiparous Holstein cows were used in a randomized complete block design with 2×2 factorial arrangement of Met (Smartamine M, Adisseo NA, Alpharetta, GA) and CHOL (ReaShure, Balchem Inc., New Hampton, NY) level (with or without). Treatments (12 cows each) were control (CON), no Met or CHOL; CON and Met (SMA); CON and CHOL (REA); and CON and Met and CHOL (MIX). From -50 to -21d before expected calving, all cows received the same diet [1.40Mcal of net energy for lactation (NEL)/kg of DM] with no Met or CHOL. From -21d to calving, cows received the same close-up diet (1.52Mcal of NEL/kg of DM) and were assigned randomly to each treatment. From calving to 30d, cows were on the same postpartal diet (1.71Mcal of NEL/kg of DM) and continued to receive the same treatments until 30d. The Met supplementation was adjusted daily at 0.08% DM of diet, and CHOL was supplemented at 60g/cow per day. Liver (-10, 7, 21, and 30d) and blood (-10, 4, 8, 20, and 30d) samples were harvested for biomarker analyses. Neutrophil and monocyte phagocytosis and oxidative burst were assessed at d 1, 4, 14, and 28d. The Met-supplemented cows tended to have greater plasma paraoxonase. Greater plasma albumin and IL-6 as well as a tendency for lower haptoglobin were detected in Met- but not CHOL-supplemented cows. Similarly, cows fed Met compared with CHOL had greater concentrations of total and reduced glutathione (a potent intracellular antioxidant) in liver tissue. Upon a pathogen challenge in vitro, blood polymorphonuclear leukocyte phagocytosis capacity and oxidative burst activity were greater in Met-supplemented cows. Overall, liver and blood biomarker analyses revealed favorable changes in liver function, inflammation status, and immune response in Met-supplemented cows

PROFILO LATTOPROTEICO E ATTIVITÀ PROTEASICA TOTALE DEL LATTE DI ASINA

RIASSUNTO - È stato condotto uno studio per valutare la variabilità del profilo lattoproteico e dell’attività proteasica totale in campioni individuali di latte d’asina di razza Amiatina. Sono stati analizzati 97 campioni di latte individuale di 26 asine prelevate più volte nel corso della lattazione. L’attività proteasica totale è stata analizzata mediante metodo colorimetrico. Sono state inoltre analizzate le lattoproteine mediante isoelectrofocusing (IEF) di campioni di latte intero. I gel sono stati successivamente quantificati mediante analisi di immagine. L’analisi IEF ha permesso di identificare un polimorfismo genetico a livello della beta-lattoglobulina (LG) I. Le varianti osservate, denominate A e B, presentavano rispettivamente una frequenza di 0,15 e 0,85 nel campione analizzato. L’attività proteasica è risultata particolarmente ridotta; questo fatto può essere imputabile al valore molto elevato di lisozima, antibatterico naturale di cui il latte d’asina è particolarmente ricco. La ripetibilità varia da un valore minimo di 0,29 (attività proteasica totale) ad un massimo di 0,69 (beta-LG). Anche per il lisozima si osserva una ripetibilità superiore al 55%. La ridotta attività proteasica dei campioni di latte analizzati e l’elevato contenuto in lisozima confermano le particolari caratteristiche nutraceutiche del latte di asina. L’analisi IEF rappresenta un’analisi a basso costo che fornisce numerose indicazioni, permettendo di mettere in luce polimorfismi genetici, come pure di quantificare il contenuto percentuale di frazioni lattoproteiche. I valori di ripetibilità osservati suggeriscono buone possibilità per la selezione di particolari frazioni sieroproteiche. Parole chiave: lattoproteine, asina, attività proteasic

The human-associated bacterium Propionibacterium acnes as a grapevine endophyte

Animals and plants have established a long-lasting cohabitation with a variety of microbes, including pathogens, commensals and beneficials. Studies investigating such associations documented numerous cases of bacterial host switches (usually from domestic animals to humans). The exchange of microbial symbionts between humans and plants is much less investigated. We reported a surprising example of horizontal interkingdom transfer of the human opportunistic pathogen (P. acnes) to grapevine (Vitis vinifera L.). P. acnes was interestingly most common inside the plant's pith tissue. Phylogenetic and population analyses place that the establishment of the grapevine-associated P. acnes likely during the Neolithic, when grapevine was domesticated. The endophytic subspecies of P. acnes was named P. Zappae. An analysis of Propionibacteria in the grapevine endosphere showed that P. Zappae is far from being the only species present in this plant as an endophyt

Detecting β-Casein Variation in Bovine Milk

In bovine species, β-casein (β-CN) is characterized by genetic polymorphism. The two most common protein variants are β-CN A2 (the original one) and A1, differing from A2 for one amino acid substitution (Pro67 to His67). Several bioactive peptides affecting milk nutritional properties can originate from β-CN. Among them, β-casomorphin-7 (BCM7) ranging from amino acid 60 to 66 can be released more easily from β-CN variants carrying His67 (A1 type) instead of Pro67 (A2 type). Nowadays, “A2 milk” is produced in different countries claiming its potential benefits in human health. The aim of this study was to further develop and apply an isoelectric focusing electrophoresis (IEF) method to bulk and individual milk samples in order to improve its use for β-CN studies. We succeeded in identifying A2 milk samples correctly and quantifying the percentage of A2, A1, and B variants in bulk samples not derived from A2 milk as well as in individual milk samples. The method allows us to quantify the relative proportion of β-CN variants in whole milk without eliminating whey protein by acid or enzymatic precipitation of caseins. The aim of this study was also to study the different behavior of β-CN and β-lactoglobulin (β-LG) in the presence of trichloroacetic acid (TCA). The higher sensitivity of β-CN to TCA allows quantifying β-CN variants after TCA fixation because β-LG is not visible. Monitoring β-CN variation in cattle breeds is important in order to maintain a certain balance between Pro67 and His67 in dairy products. Overall, the debate between A1 and A2 milk needs further investigation

Innate immune responses induced by lipopolysaccharide and lipoteichoic acid in primary goat mammary epithelial cells

Abstract Background Innate immune responses induced by in vitro stimulation of primary mammary epithelial cells (MEC) using Gram-negative lipopolysaccharide (LPS) and Gram-positive lipoteichoic acid (LTA) bacterial cell wall components are well- characterized in bovine species. The objective of the current study was to characterize the downstream regulation of the inflammatory response induced by Toll-like receptors in primary goat MEC (pgMEC). We performed quantitative real-time RT-PCR (qPCR) to measure mRNA levels of 9 genes involved in transcriptional regulation or antibacterial activity: Toll-like receptor 2 (TLR2), Toll-like receptor 4 (TLR4), prostaglandin-endoperoxide synthase 2 (PTGS2), interferon induced protein with tetratricopeptide repeats 3 (IFIT3), interferon regulatory factor 3 (IRF3), myeloid differentiation primary response 88 (MYD88), nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 (NFKB1), Toll interacting protein (TOLLIP), and lactoferrin (LTF). Furthermore, we analyzed 7 cytokines involved in Toll-like receptor signaling pathways: C-C motif chemokine ligand 2 (CCL2), C-C motif chemokine ligand 5 (CCL5), C-X-C motif chemokine ligand 6 (CXCL6), interleukin 8 (CXCL8), interleukin 1 beta (IL1B), interleukin 6 (IL6), and tumor necrosis factor alpha (TNF). Results Stimulation of pgMEC with LPS for 3 h led to an increase in expression of CCL2, CXCL6, IL6, CXCL8, PTGS2, IFIT3, MYD88, NFKB1, and TLR4 (P < 0.05). Except for IL6, and PTGS2, the same genes had greater expression than controls at 6 h post-LPS (P < 0.05). Expression of CCL5, PTGS2, IFIT3, NFKB1, TLR4, and TOLLIP was greater than controls after 3 h of incubation with LTA (P < 0.05). Compared to controls, stimulation with LTA for 6 h led to greater expression of PTGS2, IFIT3, NFKB1, and TOLLIP (P < 0.05) whereas the expression of CXCL6, CXCL8, and TLR4 was lower (P < 0.05). At 3 h incubation with both toxins compared to controls a greater expression (P < 0.05) of CCL2, CCL5, CXCL6, CXCL8, IL6, PTGS2, IFIT3, IRF3, MYD88, and NFKB1 was detected. After 6 h of incubation with both toxins, the expression of CCL2, CXCL6, IFIT3, MYD88, NFKB1, and TLR4 was higher than the controls (P < 0.05). Conclusions Data indicate that in the goat MEC, LTA induces a weaker inflammatory response than LPS. This may be related to the observation that gram-positive bacteria cause chronic mastitis more often than gram-negative infections

Additional file 1: of Innate immune responses induced by lipopolysaccharide and lipoteichoic acid in primary goat mammary epithelial cells

Additional materials. RNA extraction, purification, and quality assessment; selection of genes, primer design, quantitative RT-PCR, Table S1. Genes analyzed by quantitative PCR, and Table S2. Oligonucleotide primer sequences. (DOCX 31 kb