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 Erratum: Fukuyama, K., et al. 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, 9, 410

Pathogens ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 574
Author(s):  
Kohtaro Fukuyama ◽  
Md. Aminul Islam ◽  
Michihiro Takagi ◽  
Wakako Ikeda-Ohtsubo ◽  
Shoichiro Kurata ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Mammary Epithelial Cells ◽  
Feedlot Cattle ◽  
In Vitro Assay ◽  
Mammary Epithelial ◽  
Vitro Assay ◽  
Bovine Mammary Epithelial Cells ◽  
Published Paper

The authors would like to make the following corrections about the published paper [...]

Exogenous phospholipase A2 affects inflammatory gene expression in primary bovine mammary epithelial cells

2019 ◽  
Vol 86 (2) ◽  
pp. 177-180
Author(s):  
Jacqueline P. Kurz ◽  
Mark P. Richards ◽  
Matthew Garcia ◽  
Zhongde Wang
Keyword(s):  
Epithelial Cells ◽  
Phospholipase A2 ◽  
Inflammatory Responses ◽  
Mammary Epithelial Cells ◽  
Constitutive Expression ◽  
Mammary Epithelial ◽  
Inflammatory Factors ◽  
Bovine Mammary Epithelial Cells ◽  
Lps Challenge

AbstractThis Research Communication addresses the hypothesis that exogenously administered phospholipase A2 (PLA2) affects the inflammatory responses of bovine mammary epithelial cells (bMEC) in vitro with the aim of providing preliminary justification of investigation into the uses of exogenously administered PLA2 to manage or treat bovine mastitis. Primary bMEC lines from 11 lactating Holstein dairy cows were established and the expression of 14 pro-inflammatory genes compared under unchallenged and lipopolysaccharide (LPS)-challenged conditions, with and without concurrent treatment with bovine pancreatic PLA2G1B, a secreted form of PLA2. No differences in the expression of these genes were noted between PLA2-treated and untreated bMEC under unchallenged conditions. Following LPS challenge, untreated bMEC exhibited significant downregulation of CXCL8, IL1B, CCL20, and CXCL1. In contrast, PLA2-treated bMEC exhibited significant downregulation of IL1B and CCL20 only. These findings indicate that exogenous PLA2 affects the expression of some pro-inflammatory factors in immune-stimulated bMEC, but does not influence the constitutive expression of these factors. Further investigation of the influence of exogenous PLA2 in the bovine mammary gland is justified.

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

scholarly journals Transcriptome Analysis of The Inflammatory Responses of Bovine Mammary Epithelial Cells: Exploring Immunomodulatory Target Genes for Bovine Mastitis

Pathogens ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 200 ◽  
Author(s):  
Md. Aminul Islam ◽  
Michihiro Takagi ◽  
Kohtaro Fukuyama ◽  
Ryoya Komatsu ◽  
Leonardo Albarracin ◽  
...  
Keyword(s):  
Dairy Cows ◽  
Bacterial Infections ◽  
Target Genes ◽  
Inflammatory Responses ◽  
Bovine Mastitis ◽  
Immune Defense ◽  
Mammary Epithelial ◽  
Inflammatory Factors ◽  
Cytokines And Chemokines

Bovine mastitis is the inflammatory reaction of the mammary gland and is commonly caused by bacterial infections in high-yielding dairy cows. The detailed investigation of the immunotranscriptomic response of bovine mammary epithelial (BME) cells to pattern recognition receptors (PRRs) activation by microbial-associated molecular patterns (MAMPs) can be of great importance for understanding the innate immune defense mechanisms, and for exploring the immunomodulatory candidate genes. In this work, we investigated the transcriptome modifications of BME cells after the in vitro stimulation with Escherichia coli derived lipopolysaccharide (LPS) and heat-killed Staphylococcus aureus JE2 and S. aureus SA003. In addition, the effect of Pam3CSK4 (a synthetic triacylated lipopeptide that activates Toll-like receptor 2 (TLR2)), and the intracellular chemotactic protein cyclophilin A (CyPA), which is secreted by BME cells during mastitis, in the expression changes of selected cytokines and chemokines were evaluated by qPCR. Microarray analysis identified 447, 465 and 520 differentially expressed genes (DEGs) in the BME cells after LPS, S. aureus JE2 and S. aureus SA003 stimulation, respectively. A major differential response in the inflammatory gene expression was noticed between the stimulation of LPS and S. aureus strains. Unlike the S. aureus strains, LPS stimulation resulted in significant upregulation of CCL2, CXCL2, CXCL3, CXCL8, IL1α and IL1β, which were confirmed by qPCR analysis. Pam3CSK4 was not able to induce significant changes in the expression of cytokines and chemokines in challenged BME cells. The exogenous CyPA administration was able to upregulate CXCL2, CXCL3, CXCL8, IL1α and IL1β expression in BME cells indicating its ability to promote inflammation. The identification of transcriptional markers of mastitis specific for individual inflammatory factors such as LPS, Pam3CSK4 or CyPA, which can be evaluated in vitro in BME cells, may enable the development of novel diagnostics and/or immunomodulatory treatments, providing new tools for the effective management of mastitis in dairy cows. The results of this work are an advance in this regard.

Antagonism of Helicobacter pylori by Bacteriocins of Lactic Acid Bacteria

2003 ◽  
Vol 66 (1) ◽  
pp. 3-12 ◽  
Author(s):  
TAE-SEOK KIM ◽  
JI-WOON HUR ◽  
MYEONG-AE YU ◽  
CHAN-ICK CHEIGH ◽  
KYUNG-NAM KIM ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Helicobacter Pylori ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Inoculum Size ◽  
Strain Atcc ◽  
Initial Inoculum ◽  
H Pylori ◽  
Strain Dependent

Antimicrobial activity of seven bacteriocins produced by lactic acid bacteria against Helicobacter pylori strains (ATCC 43504, Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH [DSM] 4867, DSM 9691, and DSM 10242) was investigated in vitro using a broth microdilution assay. The bacteriocins chosen for the study were nisin A; lacticins A164, BH5, JW3, and NK24; pediocin PO2; and leucocin K. Antimicrobial activity of the bacteriocins varied among the H. pylori strains tested, of which strain ATCC 43504 was the most tolerant. Among the bacteriocins tested, lacticins A164 and BH5 produced by Lactococcus lactis subsp. lactis A164 and L. lactis BH5, respectively, showed the strongest antibacterial activity against H. pylori strains. MICs of the lacticins against H. pylori strains, when assessed by the critical dilution micromethod, ranged from 0.097 to 0.390 mg/liter (DSM strains) or from 12.5 to 25 mg/liter (ATCC 43504), supporting the strain-dependent sensitivity of the pathogen. Pediocin PO2 was less active than the lacticins against four strains of H. pylori, and leucocin K was the least active peptide, with no inhibition toward H. pylori ATCC 43504. Anti-Helicobacter activity of lacticin A164 was dependent on initial inoculum size as well as concentration of the bacteriocin added.

scholarly journals Improving Phenolic-Linked Antioxidant, Antihyperglycemic and Antibacterial Properties of Emmer and Conventional Wheat Using Beneficial Lactic Acid Bacteria

2021 ◽  
Vol 1 (2) ◽  
pp. 270-288
Author(s):  
Ashish Christopher ◽  
Dipayan Sarkar ◽  
Kalidas Shetty
Keyword(s):  
Antimicrobial Activity ◽  
Antioxidant Activity ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Pathogenic Bacteria ◽  
Phenolic Content ◽  
Bifidobacterium Longum ◽  
Antibacterial Properties ◽  
Vitro Assay

Beneficial lactic acid bacteria (LAB)-based fermentation is an effective bioprocessing approach to improve human-health-targeted functional benefits of plant-based food substrates, such as cereal grains. Previously, we observed high phenolic bioactive-linked antioxidant and anti-hyperglycemic properties in whole grain Emmer (hulled). In this study, beneficial LAB (Lactiplantibacillus plantarum) was recruited to ferment (0–72 h) aqueous extracts (0.4 g/mL concentration) of previously optimized hulled Emmer wheat and conventional red spring wheat cv. Barlow. The fermented and unfermented (control) wheat extracts were analyzed for phenolic content, phenolic profile, antioxidant activity, and antihyperglycemic properties (α-amylase and α-glucosidase enzyme inhibitory activity) using in vitro assay models. Additionally, antimicrobial activity against pathogenic bacteria Helicobacter pylori, and potential prebiotic activity supporting the growth of beneficial Bifidobacterium longum were also investigated. Improvement in antioxidant activity and antihyperglycemic functional benefits were observed, while soluble phenolic content remained high after 72 h fermentation. Antimicrobial activity against H. pylori was also observed in 48 and 72 h fermented wheat extracts. This study provides an insight into the efficacy of LAB-based fermentation as a safe bioprocessing tool to design health-targeted functional foods and ingredients from underutilized whole grains like Emmer for targeting type 2 diabetes dietary benefits.

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.

scholarly journals Adjunct Culture of Non-Starter Lactic Acid Bacteria for the Production of Provola Dei Nebrodi PDO Cheese: In Vitro Screening and Pilot-Scale Cheese-Making

2021 ◽  
Vol 9 (1) ◽  
pp. 179
Author(s):  
Cinzia Lucia Randazzo ◽  
Luigi Liotta ◽  
Maria De Angelis ◽  
Giuseppe Celano ◽  
Nunziatina Russo ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Pilot Scale ◽  
Cheese Making ◽  
Flight Mass Spectrometry ◽  
Volatile Organic ◽  
Strain Dependent ◽  
Cheese Production ◽  
Adjunct Culture

The present study aimed at selecting non-starter lactic acid bacteria strains, with desirable technological and enzymatic activities, suitable as adjunct culture for the Provola dei Nebrodi cheese production. One hundred and twenty-one lactic acid bacteria, isolated from traditional Provola dei Nebrodi cheese samples, were genetically identified by Rep-PCR genomic fingerprinting, using the (GTG)5-primer, and by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS). Twenty-seven strains, included in the qualified presumption of safety (QPS) list, were tested for technological and proteinase/peptidase activities. Results showed that technological features and flavour formation abilities were strain-dependent. Among the selected strains, Lacticaseibacillus paracasei PN 76 and Limosilactobacillus fermentum PN 101 were used as adjunct culture in pilot-scale cheese-making trials. Data revealed that adjunct cultures positively affected the flavour development of cheese, starting from 30 days of ripening, contributing to the formation of key flavour compounds. The volatile organic compound profiles of experimental cheeses was significantly different from those generated in the controls, suggesting that the selected adjunct strains were able to accelerate the flavour development, contributing to a unique profile of Provola dei Nebrodi cheese.

scholarly journals Improving Phenolic Bioactive-Linked Functional Qualities of Sweet Potatoes Using Beneficial Lactic Acid Bacteria-Based Biotransformation Strategy

Horticulturae ◽  
2021 ◽  
Vol 7 (10) ◽  
pp. 367
Author(s):  
Pradeepika Chintha ◽  
Dipayan Sarkar ◽  
Kenneth Pecota ◽  
Munevver Dogramaci ◽  
Kalidas Shetty
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Human Health ◽  
Sweet Potato ◽  
Phenolic Content ◽  
Cold Water ◽  
Sweet Potatoes ◽  
Vitro Assay ◽  
Food Grade

Beneficial lactic acid bacteria (LAB) based fermentation is an effective biotransformation strategy to preserve and improve the human health-supporting functional qualities of plant-based food substrates. In this study, a food grade strain of Lactiplantibacillus plantarum was recruited to improve the retention, stability, and bioavailability of phenolic bioactives to enhance the antioxidant, anti-hyperglycemic, and anti-hypertensive functional qualities of three flesh-colored sweet potato varieties, Murasaki (off-white-fleshed), Evangeline (orange-fleshed), and NIC-413 (purple-fleshed). Liquid (cold water) extracts of the sweet potatoes, which are relevant for food grade applications, were fermented for 72 h at 37 °C. Total soluble phenolic content, phenolic profile, antioxidant, anti-hyperglycemic, and anti-hypertensive benefits relevant functional properties of fermented and unfermented sweet potato extracts were evaluated at 0, 24, 48, and 72 h time points using in vitro assay models. Overall, high total soluble phenolic content and total antioxidant activity were observed at 24 h, retaining this high level even after 72 h of fermentation. Additionally, moderate to high α-amylase, α-glucosidase, and angiotensin-I-converting enzyme inhibitory activities were observed in the fermented sweet potato extracts. The results suggested that LAB-based fermentation is an effective post-harvest processing strategy for a higher retention of phenolic bioactives and concurrently improves the human health protective bioactive functional qualities of sweet potatoes.

scholarly journals The Protective Effects of Lactobacillus plantarum KLDS 1.0344 on LPS-Induced Mastitis In Vitro and In Vivo

2021 ◽  
Vol 12 ◽  
Author(s):  
Qingxue Chen ◽  
Song Wang ◽  
Jiayao Guo ◽  
Qinggang Xie ◽  
Smith Etareri Evivie ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Pathogenic Bacteria ◽  
Mammary Epithelial ◽  
Inflammatory Factors ◽  
Protective Effects ◽  
E Coli ◽  
Tnf Α

Cow mastitis, which significantly lowers milk quality, is mainly caused by pathogenic bacteria such as E. coli. Previous studies have suggested that lactic acid bacteria can have antagonistic effects on pathogenic bacteria that cause mastitis. In the current study, we evaluated the in vitro and in vivo alleviative effects of L. plantarum KLDS 1.0344 in mastitis treatment. In vitro antibacterial experiments were performed using bovine mammary epithelial cell (bMEC), followed by in vivo studies involving mastitis mouse models. In vitro results indicate that lactic acid was the primary substance inhibiting the E. coli pathogen. Meanwhile, treatment with L. plantarum KLDS 1.0344 can reduce cytokines’ mRNA expression levels in the inflammatory response of bMEC induced by LPS. In vivo, the use of this strain reduced the secretion of inflammatory factors IL-6, IL-1β, and TNF-α, and decreased the activity of myeloperoxidase (MPO), and inhibited the secretion of p-p65 and p-IκBα. These results indicate that L. plantarum KLDS 1.0344 pretreatment can reduce the expression of inflammatory factors by inhibiting the activation of NF-κB signaling pathway, thus exerting prevent the occurrence of inflammation in vivo. Our findings show that L. plantarum KLDS 1.0344 has excellent properties as an alternative to antibiotics and can be developed into lactic acid bacteria preparation to prevent mastitis disease.

Sign in / Sign up

Export Citation Format

Share Document