Bovine mastitis prevention: humoral and cellular response of dairy cows inoculated with lactic acid bacteria at the dry-off period

2017 ◽  
Vol 8 (4) ◽  
pp. 589-596 ◽  
Author(s):  
M. Pellegrino ◽  
N. Berardo ◽  
J. Giraudo ◽  
M.E.F. Nader-Macías ◽  
C. Bogni
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Cellular Response ◽  
Animal Feed ◽  
Bovine Milk ◽  
Bovine Mastitis ◽  
Memory Cell ◽  
Microbiological Analysis ◽  
Mastitis Pathogens

The use of lactic acid bacteria (LAB) in animal feed, constitute an alternative tool for bovine mastitis prevention. Previously, two LAB strains were isolated from bovine milk and selected for their probiotics properties. So far, immune response of inoculating LAB in bovine udders at dry-off period has not been investigated. The immunoglobulin isotype levels and memory cell proliferation in blood and milk of animals inoculated with Lactobacillus lactis subsp. lactis CRL1655 and Lactobacillus perolens CRL1724 at dry-off period was studied. Ten animals were inoculated intramammarily with 106 cells of each LAB (IG) and 2 animals used as control (NIG). Milk and blood samples were taken before inoculation and 1, 2, 4, 6, 12 and 24 h and 7 and 14 days after inoculation. Somatic cell count (SCC) in milk, the presence of bovine mastitis pathogens, the levels of antibodies and lymphocyte proliferation were determined. In the IG, the SCC was <250,000 cells/ml up to 4 h after intramammary inoculation. Six and 12 h after inoculation, the SCC increased up to 600,000 and 2,000,000 cells/ml, respectively. In the NIG, the SCC reached the maximum value 7 days after inoculation. Microbiological analysis showed that all samples were negative for major bovine mastitis pathogens after 24-48 h of incubation. In general, LAB inoculation increased the amount of IgG isotypes in blood and milk, and these antibodies were able to recognise Staphylococcus aureus epitopes. Lymphocytes proliferation was significantly higher in the IG at all time points assayed, following LAB or S. aureus stimulation. The lymphocytes of animals inoculated with LAB do not react in vitro to the presence of S. aureus antigen.. The results showed that probiotic microorganisms could be a natural and effective alternative in the prevention of bovine mastitis at dry-off period and act as immunomodulatory stimulating local and systemic defence lines.

Intramammary inoculation with lactic acid bacteria at dry-off triggers an immunomodulatory response in dairy cows

2020 ◽  
Vol 11 (6) ◽  
pp. 561-572
Author(s):  
N. Berardo ◽  
L. Bohl ◽  
C. Porporatto ◽  
M.E.F. Nader-Macias ◽  
C. Bogni ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Blood Serum ◽  
Cellular Response ◽  
Animal Feed ◽  
Bovine Milk ◽  
Bovine Mastitis ◽  
Post Inoculation ◽  
Streptococcus Uberis ◽  
Significant Difference

The use of antibiotics to prevent bovine mastitis is responsible for the emergence and selection of resistant strains. Lactic acid bacteria (LAB) could be introduced into animal feed as an alternative prevention method that would bypass the risk of resistance development. In previous research, we demonstrated that two probiotic LAB strains isolated from bovine milk were capable of stimulating the production of antibodies and the host’s immune cellular response in the udder. The present study aimed to elucidate whether the antibodies of animals inoculated with these strains were able to increase phagocytosis by neutrophils and inhibit the growth of different mastitis-causing pathogens. Moreover, the effect of LAB on the expression of pro-inflammatory cytokines was assessed. Ten animals were inoculated intramammarily with 106 cells of the two strains at dry-off. The blood serum was tested for its ability to opsonize bovine mastitis pathogens, the in vitro bactericidal activity of bovine blood and milk against these pathogens was determined, and cytokine mRNA expression was quantified in milk somatic cells. The inoculated animals did not show abnormal signs of sensitivity to the LAB. Their blood serum significantly enhanced the phagocytosis of Staphylococcus spp. and the LAB. Escherichia coli and Streptococcus uberis were inhibited by the milk serum but not the blood serum, whereas Staphylococcus aureus and Staphylococcus haemolyticus were inhibited by both. In regard to cytokine expression, interleukin (IL)-1β increased markedly for up to 4 h post-inoculation, and an increase in IL-8 was observed 4, 12 and 24 h after inoculation. Tumour necrosis factor-α mRNA increased 1 and 2 h after inoculation and a significant difference was registered at 6 h for interferon-γ. This rapid immunomodulatory response shows that inoculating animals with LAB at dry-off, when they are especially susceptible, could be a useful strategy for the prevention of bovine mastitis.

In Vitro Characterization of Lactic Acid Bacteria Isolated from Bovine Milk as Potential Probiotic Strains to Prevent Bovine Mastitis

2018 ◽  
Vol 11 (1) ◽  
pp. 74-84 ◽  
Author(s):  
Matías S. Pellegrino ◽  
Ignacio D. Frola ◽  
Berardo Natanael ◽  
Dino Gobelli ◽  
María E.F. Nader-Macias ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Bovine Milk ◽  
Bovine Mastitis ◽  
In Vitro Characterization ◽  
Probiotic Strains

Efficacy of purified antimicrobial peptides from lactic acid bacteria against bovine mastitis pathogens

2015 ◽  
Vol 34 (4) ◽  
Author(s):  
R. Geetha ◽  
C. T. Sathian ◽  
V. Prasad ◽  
V. L. Gleeja
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Antimicrobial Peptides ◽  
Reference Strain ◽  
Bovine Mastitis ◽  
Zone Of Inhibition ◽  
Salting Out ◽  
Inhibitory Spectrum ◽  
Inhibitory Efficiency ◽  
Mastitis Pathogens

A study was carried out to assess the efficacy of purified antimicrobial peptides of lactic acid bacteria (LAB), isolated from partially decayed samples of food items, against selected clinical isolates of mastitis bacterial pathogens, <italic>viz</italic>., <italic>Escherichia coli</italic> (<italic>E.coli</italic>), <italic>Staphylococcus aureus</italic> (<italic>S.aureus</italic>) and <italic>Streptococcus agalactiae</italic> (<italic>S.agalactiae</italic>). The crude culture extracts (CCE) from LAB isolates were concentrated and purified by salting out method followed by dialysis. Partially purified peptides (T-5 CR -2 CR -3) from three LAB isolated from tomato and carrot showed a significantly higher inhibitory spectrum, with zone of inhibition ranging from 11.66±0.33 to 12.66±0.33 mm diameter against different test pathogens. The inhibitory efficiency of all the three peptides was found to be significantly superior to the peptides from reference strain <italic>Lactococcus lactis</italic> NCIM-2114. Minimum inhibitory concentration (MIC) of the purified peptide samples ranged between 0.79±0.09 to 1.11±0.09 μg/ml against different test pathogens. The activity of peptides was completely lost after treatment with protease, thereby confining their protein character.

scholarly journals Evaluation of the Immunomodulatory Ability of Lactic Acid Bacteria Isolated from Feedlot Cattle Against Mastitis Using a Bovine Mammary Epithelial Cells In Vitro Assay

Pathogens ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 410 ◽  
Author(s):  
Kohtaro Fukuyama ◽  
Md. Aminul Islam ◽  
Michihiro Takagi ◽  
Wakako Ikeda-Ohtsubo ◽  
Shoichiro Kurata ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Inflammatory Responses ◽  
Interleukin 1 ◽  
Bovine Mastitis ◽  
Mammary Epithelial ◽  
Vitro Assay ◽  
Lps Challenge ◽  
Strain Dependent

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.

scholarly journals In vitro efficacy of teat antiseptics against Staphylococcus aureus strains isolated from bovine mastitis

2016 ◽  
Vol 37 (4) ◽  
pp. 1997
Author(s):  
Renata Paoli Santos ◽  
Fernando Nogueira de Souza ◽  
Carla Gasparotto Chande Vasconcelos ◽  
Adriana Cortez ◽  
Dalila Lapinha Silva Oliveira Rosa ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Lactic Acid ◽  
Bovine Mastitis ◽  
Resistance Profile ◽  
Intramammary Infections ◽  
Time Points ◽  
Control Programs ◽  
Mastitis Pathogens ◽  
Mastitis Control

The process of teat disinfection is a widely accepted component of successful mastitis control programs by reducing the number of bacteria on the teat skin and healing teat lesions. For contagious pathogens such as Staphylococcus aureus, post-milking teat disinfection remains a simple, effective and economical practice for prevention of new intramammary infections (IMIs) of lactating dairy cows. Despite the universal acceptance of teat dipping as a method of mastitis control, variations in the susceptibility and resistance profile of mastitis pathogens among antiseptics have been described. Thus, here we sought to explore the in vitro efficacy of the followings antiseptics against S. aureus isolated from IMIs: chlorhexidine (2.0%), chlorine (2.5%), quarternary ammonium (4.0%), lactic acid (2.0%) and iodine (0.6%). We used 50 S. aureus strains isolated from bovine IMIs from 50 dairy herds located at Minas Gerais, São Paulo, Paraná and Rio Grande do Sul States (Brazil). The antiseptics were evaluated at four different specific intervals (15, 30, 60 and 300 s). We found a higher activity for quarternary ammonium and chlorhexidine against S. aureus at all time-points, followed by iodine and then chlorine. Lactic acid treatment produced the worst results for all time-points and strains. Due to variations in the sensitivity and resistance profile of antiseptics against S. aureus isolated from IMIs, the effectiveness of the antiseptics against the major mastitis pathogens should be periodically evaluated in dairy farms in an attempt to reduce the rate of new IMIs in the herd.

scholarly journals Function-Driven Design of Lactic Acid Bacteria Co-cultures to Produce New Fermented Food Associating Milk and Lupin

2020 ◽  
Vol 11 ◽  
Author(s):  
Fanny Canon ◽  
Mahendra Mariadassou ◽  
Marie-Bernadette Maillard ◽  
Hélène Falentin ◽  
Sandrine Parayre ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Bovine Milk ◽  
Fermented Food ◽  
Positive Interactions ◽  
Fermented Foods ◽  
Raffinose Family Oligosaccharides ◽  
Cell Counts ◽  
Carbohydrate Degradation

Designing bacterial co-cultures adapted to ferment mixes of vegetal and animal resources for food diversification and sustainability is becoming a challenge. Among bacteria used in food fermentation, lactic acid bacteria (LAB) are good candidates, as they are used as starter or adjunct in numerous fermented foods, where they allow preservation, enhanced digestibility, and improved flavor. We developed here a strategy to design LAB co-cultures able to ferment a new food made of bovine milk and lupin flour, consisting in: (i) in silico preselection of LAB species for targeted carbohydrate degradation; (ii) in vitro screening of 97 strains of the selected species for their ability to ferment carbohydrates and hydrolyze proteins from milk and lupin and clustering strains that displayed similar phenotypes; and (iii) assembling strains randomly sampled from clusters that showed complementary phenotypes. The designed co-cultures successfully expressed the targeted traits i.e., hydrolyzed proteins and degraded raffinose family oligosaccharides of lupin and lactose of milk in a large range of concentrations. They also reduced an off-flavor-generating volatile, hexanal, and produced various desirable flavor compounds. Most of the strains in co-cultures achieved higher cell counts than in monoculture, suggesting positive interactions. This work opens new avenues for the development of innovative fermented food products based on functionally complementary strains in the world-wide context of diet diversification.

scholarly journals Short communication: Lactic acid bacteria from the honeybee inhibit the in vitro growth of mastitis pathogens

2016 ◽  
Vol 99 (4) ◽  
pp. 2940-2944 ◽  
Author(s):  
K. Piccart ◽  
A. Vásquez ◽  
S. Piepers ◽  
S. De Vliegher ◽  
T.C. Olofsson
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Short Communication ◽  
In Vitro Growth ◽  
Mastitis Pathogens

In vitro adhesion of lactic acid bacteria and bifidobacteria to Caco-2P and IEC-18 cells

2005 ◽  
Vol 34 (1) ◽  
pp. 91-99 ◽  
Author(s):  
K. Szekér ◽  
J. Beczner ◽  
A. Halász ◽  
Á. Mayer ◽  
J.M. Rezessy-Szabó ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
In Vitro Adhesion

scholarly journals Influence of nutrient availability on in vitro growth of major bovine mastitis pathogens

2021 ◽  
Vol 88 (1) ◽  
pp. 80-88
Author(s):  
Remo Stürmlin ◽  
Josef J. Gross ◽  
Olga Wellnitz ◽  
Lea A. Wagner ◽  
Camille Monney ◽  
...  
Keyword(s):  
Bacterial Growth ◽  
Nutrient Availability ◽  
Bovine Mastitis ◽  
Milk Composition ◽  
B Vitamins ◽  
E Coli ◽  
Streptococcus Uberis ◽  
Whole Milk ◽  
Mastitis Pathogens

AbstractThe aim of the present study was to investigate the effects of milk composition changes on the in vitro growth of bovine mastitis pathogens. Nutritional requirements of three major bovine mastitis pathogens Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), and Streptococcus uberis (S. uberis) were investigated in vitro. We used ultra-high temperature (UHT) treated milk with different contents of fat, protein, and carbohydrates to test the influence of the availability of various milk constituents on pathogen growth characteristics. Additionally, the bacterial growth was investigated under experimentally modified nutrient availability by dilution and subsequent supplementation with individual nutrients (carbohydrates, different nitrogen sources, minerals, and different types of B vitamins) either to milk or to a conventional medium (thioglycolate broth, TB). Varying contents of fat, protein or lactose did not affect bacterial growth with the exception of growth of S. uberis being promoted in protein-enriched milk. The addition of nutrients to diluted whole milk and TB partly revealed different effects, indicating that there are media-specific growth limiting factors after dilution. Supplementation of minerals to diluted milk did not affect growth rates of all studied bacteria. Bacterial growth in diluted whole milk was decreased by the addition of high concentrations of amino acids in S. aureus, and by urea and additional B vitamins in E. coli and S. aureus. The growth rate of S. uberis was increased by the addition of B vitamins to diluted whole milk. The present results demonstrate that growth-limiting nutrients differ among pathogen types. Because reduced bacterial growth was only shown in diluted milk or TB, it is unlikely that alterations in nutrient availability occurring as a consequence of physiological changes of milk composition in the cow's udder would directly affect the susceptibility or course of bovine mastitis.

scholarly journals Improving ensiling characteristics by adding lactic acid bacteria modifies in vitro digestibility and methane production of forage-sorghum mixture silage

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Chatchai Kaewpila ◽  
Pongsatorn Gunun ◽  
Piyawit Kesorn ◽  
Sayan Subepang ◽  
Suwit Thip-uten ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Nutritive Value ◽  
Gas Production ◽  
In Vitro Digestibility ◽  
Acid Fermentation ◽  
Design Factor ◽  
Forage Sorghum ◽  
Randomized Design

AbstractImproving the nutrition of livestock is an important aspect of global food production sustainability. This study verified whether lactic acid bacteria (LAB) inoculant could promote ensiling characteristics, nutritive value, and in vitro enteric methane (CH4) mitigation of forage sorghum (FS) mixture silage in attacking malnutrition in Zebu beef cattle. The FS at the soft dough stage, Cavalcade hay (CH), and cassava chip (CC) were obtained. The treatments were designed as a 4 × 2 factorial arrangement in a completely randomized design. Factor A was FS prepared without or with CH, CC, and CH + CC. Factor B was untreated or treated with Lactobacillus casei TH14. The results showed that all FS mixture silages preserved well with lower pH values below 4.0 and higher lactic acid contents above 56.4 g/kg dry matter (DM). Adding LAB boosted the lactic acid content of silages. After 24 h and 48 h of in vitro rumen incubation, the CC-treated silage increased in vitro DM digestibility (IVDMD) with increased total gas production and CH4 production. The LAB-treated silage increased IVDMD but decreased CH4 production. Thus, the addition of L. casei TH14 inoculant could improve lactic acid fermentation, in vitro digestibility, and CH4 mitigation in the FS mixture silages.

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