Abstract
Bovine mastitis, the inflammation of the mammary gland, affects the quality and quantity of milk yield. Mastitis control relies on single or multiple combinations of antibiotic therapy. Due to increasing antibiotic resistance in pathogens, the intramammary infusion of lactic acid bacteria (LAB) has been considered as a potential alternative to antibiotics for treating and preventing bovine mastitis through the improvement of the host immunity. Probiotic effects are a strain-dependent characteristic; therefore, candidate LAB strains have to be evaluated efficiently to find out the ones with the best potential. Here, we investigated LAB strains originally isolated from feedlot cattle’s environment regarding their ability in inducing the Toll-like receptor (TLR)-triggered inflammatory responses in bovine mammary epithelial (BME) cells in vitro. The BME cells were pre-stimulated with the LAB strains individually for 12, 24, and 48 h and then challenged with Escherichia coli-derived lipopolysaccharide (LPS) for 12 h. The mRNA expression of selected immune genes—interleukin 1 alpha (IL-1α), IL-1β, monocyte chemotactic protein 1 (MCP-1), IL-8, chemokine (C-X-C motif) ligand 2 (CXCL2), and CXCL3 were quantified by real-time quantitative PCR (RT-qPCR). Results indicated that pretreatment with some Lactobacillus strains were able to differentially regulate the LPS inflammatory response in BME cells; however, strain-dependent differences were found. The most remarkable effects were found for Lactobacillus acidophilus CRL2074, which reduced the expression of IL-1α, IL-1β, MCP-1, IL-8, and CXCL3, whereas Lactobacillus rhamnosus CRL2084 diminished IL-1β, MCP-1, and IL-8 expression. The pre-stimulation of BME cells with the CRL2074 strain resulted in the upregulated expression of three negative regulators of the TLRs, including the ubiquitin-editing enzyme A20 (also called tumor necrosis factor alpha-induced protein 3, TNFAIP3), single immunoglobin IL-1 single receptor (SIGIRR), and Toll interacting protein (Tollip) after the LPS challenge. The CRL2084 pre-stimulation upregulated only Tollip expression. Our results demonstrated that the L. acidophilus CRL2074 strain possess remarkable immunomodulatory abilities against LPS-induced inflammation in BME cells. This Lactobacillus strain could be used as candidate for in vivo testing due to its beneficial effects in bovine mastitis through intramammary infusion. Our findings also suggest that the BME cells immunoassay system could be of value for the in vitro evaluation of the immunomodulatory abilities of LAB against the inflammation resulting from the intramammary infection with mastitis-related pathogens.
Highlights
Bovine mastitis is defined as the inflammation of the mammary gland, which greatly affects milk production, animal health and welfare, and economic profit of dairy worldwide [1]
Our results demonstrated that L. mucosae CRL2069 and L. rhamnosus CRL2084 had in response to infections
Dynamics of TLR2 and Toll-like receptor 4 (TLR4) in bovine mammary epithelial (BME) Cells after Ligand Stimulation cattle lactobacilli to modulate the bovine mammary gland innate immunity triggered by Escherichia coli (E. coli)-derived lipopolysaccharide (LPS) using the BME cells in vitro system
Summary
Bovine mastitis is defined as the inflammation of the mammary gland, which greatly affects milk production, animal health and welfare, and economic profit of dairy worldwide [1]. Prophylactic intramammary infusion of long-acting antibiotics is frequently practiced to prevent intramammary infection in a dry period known as “dry cow therapy” [7]. For both prophylactic and therapeutic cases, a single or a combination of multiple antibiotics can be prescribed. Because of the increased probability of transmission of antibiotic resistance genes to indigenous and potential pathogens through antibiotic therapy as well as the poor cure rates of mastitis during lactation [10,11], the conventional treatment method needs to be revisited, and innovative and sustainable therapeutic alternatives should be sought
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