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.

Efficacy of a teat dip containing the bacteriocin lacticin 3147 to eliminate Gram-positive pathogens associated with bovine mastitis

2009 ◽  
Vol 77 (2) ◽  
pp. 231-238 ◽  
Author(s):  
Katja Klostermann ◽  
Fiona Crispie ◽  
Jim Flynn ◽  
William J Meaney ◽  
R Paul Ross ◽  
...  
Keyword(s):  
Bovine Mastitis ◽  
Gram Positive ◽  
Lactating Cows ◽  
Streptococcus Uberis ◽  
Colony Forming Units ◽  
Lacticin 3147 ◽  
Mastitis Pathogens ◽  
Teat Canal

On most dairy farms teat dips are applied to the teats of cows either before or after milking in order to prevent pathogens from gaining access to the mammary gland via the teat canal. In the present experiments, a natural teat dip was developed using a fermentate containing the live bacteriumLactococcus lactisDPC 3251. This bacterium produces lacticin 3147, a two-component lantibiotic which was previously shown to effectively kill Gram-positive mastitis pathogens. Lacticin 3147 activity in the fermentate was retained at 53% of its original level following storage for 3 weeks at 4°C. In the initial experiments in vitro, 105colony-forming units/ml (cfu/ml) of eitherStaphylococcus aureus,Streptococcus dysgalactiaeorStreptococcus uberiswere introduced into the lacticin-containing fermentate. NeitherStaph. aureusnorStr. dysgalactiaecould be detected after 30 min or 15 min, respectively, whileStr. uberiswas reduced approximately 100-fold after 15 min. Following these trials, preliminary experiments were performed in vivo on teats of lactating dairy cows. In these experiments, teats were coated with each of the challenge organisms and then dipped with the lacticin-containing fermented teat dip. Following a dip contact time of 10 min, staphylococci were reduced by 80% when compared with the undipped control teat. Streptococcal challenges were reduced by 97% forStr. dysgalactiaeand by 90% forStr. uberis. These trials showed that the teat dip is able to reduce mastitis pathogens on the teats of lactating cows.

scholarly journals Reversible inhibition of bacterial growth after specific inhibition of spermidine synthase by dicyclohexylamine

1984 ◽  
Vol 223 (3) ◽  
pp. 823-830 ◽  
Author(s):  
T Mattila ◽  
T Honkanen-Buzalski ◽  
H Pösö
Keyword(s):  
Final Concentration ◽  
Spermidine Synthase ◽  
Bacterial Cells ◽  
Bacterial Strains ◽  
E Coli ◽  
Inhibition Of Growth ◽  
Streptococcus Uberis ◽  
Mastitis Pathogens

The effect of dicyclohexylamine on seven freshly isolated bacterial strains of mastitis pathogens was studied. Streptococcus uberis was the most sensitive strain investigated, since 5 mM-dicyclohexylamine totally arrested its growth and 1.25 mM of the drug caused 60% growth inhibition. The Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa strains were also sensitive to the drug, but less so than Strep. uberis, since 5 mM drug caused only partial inhibition of growth. Micrococcus sp. and Klebsiella sp. grew in the presence of 10.0 mM-dicyclohexylamine, and, finally the growth of Streptococcus agalactiae was not at all affected by dicyclohexylamine. These different sensitivities towards dicyclohexylamine in vivo were paralleled by different sensitivities of the bacteria's spermidine synthase to the drug in vitro, and also by the ability of the drug to lower spermidine concentration in bacterial cells. Spermidine synthase from sensitive bacteria was inhibited by more than 90% by 50 microM-dicyclohexylamine in vitro, and the concentration of spermidine was decreased in E. coli and Ps. aeruginosa by 70% and in Strep. uberis by 95%, whereas in Strep. agalactiae 5 mM-dicyclohexylamine did not affect the concentration of spermidine at all. Dicyclohexylamine treatment led to the accumulation of putrescine in Strep. uberis. Spermidine synthesis catalysed by the extracts of Micrococcus sp. required 500 microM-dicyclohexylamine for 90% inhibition, and Strep. agalactiae contained a spermidine synthase that was still active at 1000 microM-dicyclohexylamine, The observed inhibition of growth was totally reversed by adding 50 microM-spermidine (final concentration) to the medium. Putrescine reversed the inhibition only when bacteria had a spermidine synthase activity insensitive to dicyclohexylamine. Spermine did not overcome the inhibition of growth caused by dicyclohexylamine, probably because it was not taken up by the bacterial cells used in this study. The inhibition of the growth by dicyclohexylamine (even in the case of Strep. uberis) was reversible in the sense that addition of 50 microM-spermidine 18 h after dicyclohexylamine still restored the growth rate of untreated controls.

scholarly journals Advances in lactoferrin research concerning bovine mastitis

2017 ◽  
Vol 95 (1) ◽  
pp. 69-75 ◽  
Author(s):  
Kei-ichi Shimazaki ◽  
Kazuhiro Kawai
Keyword(s):  
Immune System ◽  
Defense Mechanism ◽  
Bovine Mastitis ◽  
Healthy Development ◽  
Treatment And Prevention ◽  
Year 2000 ◽  
Mastitis Pathogens ◽  
Binding Glycoprotein

Lactoferrin is a multifunctional, iron-binding glycoprotein found in milk and other exocrine secretions. Lactoferrin in milk plays vital roles in the healthy development of newborn mammals, and is also an innate resistance factor involved in the prevention of mammary gland infection by microorganisms. Inflammation of the udder because of bacterial infection is referred to as mastitis. There have been many investigations into the relationships between lactoferrin and mastitis, which fall into several categories. The main categories are fluctuations in the lactoferrin concentration of milk, lactoferrin activity against mastitis pathogens, elucidation of the processes underlying the onset of mastitis, participation of lactoferrin in the immune system, and utilization of lactoferrin in mastitis treatment and prevention. This minireview describes lactoferrin research concerning bovine mastitis. In the 1970s, many researchers reported that the lactoferrin concentration fluctuates in milk from cows with mastitis. From the late 1980s, many studies clarified the infection-defense mechanism in the udder and the contribution of lactoferrin to the immune system. After the year 2000, the processes underlying the onset of mastitis were elucidated in vivo and in vitro, and lactoferrin was applied for the treatment and prevention of mastitis.

Effect of Tannins on the In Vitro Growth of Escherichia coli O157:H7 and In Vivo Growth of Generic Escherichia coli Excreted from Steers

2007 ◽  
Vol 70 (3) ◽  
pp. 543-550 ◽  
Author(s):  
BYENG R. MIN ◽  
WILLIAM E. PINCHAK ◽  
ROBIN C. ANDERSON ◽  
TODD R. CALLAWAY
Keyword(s):  
Escherichia Coli ◽  
Growth Rate ◽  
Bacterial Growth ◽  
Escherichia Coli O157 ◽  
Tannin Extract ◽  
Bacterial Growth Rate ◽  
Linear Effect ◽  
E Coli

The effect of commercially available chestnut and mimosa tannins in vitro (experiment 1) or in vivo (experiment 2) on the growth or recovery of Escherichia coli O157:H7 or generic fecal E. coli was evaluated. In experiment 1, the mean growth rate of E. coli O157:H7, determined via the measurement of optical density at 600 nm during anaerobic culture in tryptic soy broth at 37°C, was reduced (P < 0.05) with as little as 400 μg of either tannin extract per ml of culture fluid. The addition of 200, 400, 600, 800, and 1,200 μg of tannins per ml significantly (P < 0.01) reduced the specific bacterial growth rate when compared with the nontannin control. The specific growth rate decreased with increasing dose levels up to 800 μg of tannins per ml. Bacterial growth inhibition effects in chestnut tannins were less pronounced than in mimosa tannins. Chestnut tannin extract addition ranged from 0 to 1,200 μg/ml, and a linear effect (P < 0.05) was observed in cultures incubated for 6 h against the recovery of viable cells, determined via the plating of each strain onto MacConkey agar, of E. coli O157:H7 strains 933 and 86-24, but not against strain 6058. Similar tests with mimosa tannin extract showed a linear effect (P < 0.05) against the recovery of E. coli O157:H7 strain 933 only. The bactericidal effect observed in cultures incubated for 24 h with the tannin preparations was similar, although it was less than that observed from cultures incubated for 6 h. When chestnut tannins (15 g of tannins per day) were infused intraruminally to steers fed a Bermuda grass hay diet in experiment 2, fecal E. coli shedding was lower on days 3 (P < 0.03), 12 (P = 0.08), and 15 (P < 0.001) when compared with animals that were fed a similar diet without tannin supplementation. It was concluded that dietary levels and sources of tannins potentially reduce the shedding of E. coli from the gastrointestinal tract.

scholarly journals In vitro antimicrobial effect of palm oils rich in medium-chain fatty acids against mastitis-causing Gram-positive bacteria

2019 ◽  
Vol 64 (No. 8) ◽  
pp. 325-331
Author(s):  
Klára Laloučková ◽  
Lucie Malá ◽  
Paula Slaničková ◽  
Eva Skřivanová
Keyword(s):  
Fatty Acids ◽  
Dairy Cattle ◽  
Antibacterial Effect ◽  
Bovine Mastitis ◽  
Medium Chain ◽  
Medium Chain Fatty Acids ◽  
Streptococcus Uberis ◽  
Broth Microdilution Method ◽  
Chain Fatty Acids

Various pathogens causing mastitis in dairy cattle are of serious concern due to their increasing antibacterial resistance and potential transmission to other cows, calves, and the environment, especially through the milking process. Therefore, alternative approaches to antimicrobial usage in the treatment or control of mastitis in dairy cattle are severely needed. The antibacterial effect of medium-chain fatty acids (MCFAs) is known to be significant for various pathogens, but there is only limited information about the activity of MCFAs on mastitis-causing pathogens. Moreover, no evidence about the antimicrobial effects of palm oils rich in MCFAs, such as coconut, palm kernel, and tucuma oil, can be found in the current literature. The aim of this study was to evaluate the in vitro antibacterial effect of palm oils rich in MCFAs, after cleavage by an exogenous lipase from Mucor javanicus, on bovine mastitis-causing strains (Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, and Streptococcus uberis) by the broth microdilution method. All tested palm oils exerted antibacterial activity against eight tested bacterial strains in the range of 64–8192 µl/ml with Str. agalactiae being the most sensitive and S. aureus being the most resistant species. The results of the present study demonstrate that palm oils rich in MCFAs can serve as an alternative to the predominantly used predip and postdip procedures in bovine mastitis control, but further in vivo studies are needed to confirm the findings for their possible applications.

scholarly journals Immune relevant gene expression of mammary epithelial cells and their influence on leukocyte chemotaxis in response to different mastitis pathogens

2012 ◽  
Vol 51 (No. 4) ◽  
pp. 125-132 ◽  
Author(s):  
O. Wellnitz ◽  
P. Reith ◽  
Haas SC ◽  
Meyer HHD
Keyword(s):  
Immune Response ◽  
Epithelial Cells ◽  
Mrna Expression ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
E Coli ◽  
Amyloid A ◽  
Streptococcus Uberis ◽  
Leukocyte Chemotaxis ◽  
Mastitis Pathogens

Different mastitis pathogens induce different courses of infection, i.e. more or less severe. Mammary epithelial cells play an important role in the initial combat against microorganisms by expression of cytokines and acute phase proteins that regulate the immune response. The objective of the present study was to investigate the involvement of the epithelial cells into the outcome of mastitis induced by different pathogens. Primary epithelial cell cultures isolated from milk were used to test the immune response by measuring the mRNA expression of immunomodulators and their influence on polymorph nuclear chemotaxis. Because the cells showed different responses to isolated bacterial endotoxins (lipopolysaccharide, lipoteichoic acid, and peptidoglycans) compared to whole bacteria, they were treated with heat inactivated (10 MOI) gram-negative Escherichia coli, a very common pathogen causing acute intra-mammary infections, with Staphylococcus aureus, a prevalent cause of chronic, and, Streptococcus uberis, an inducer of acute and chronic mastitis. E. coli induced an increased mRNA expression of interleukin (IL)-8 within a 1 h treatment. A treatment for 6 h with E. coli and S. aureus induced increased mRNA expression of IL-6, IL-8, TNF-&aacute; and serum amyloid A (SAA). After a 24 h treatment the expression of these immunomodulators was still elevated, except in the E. coli treatment the SAA expression showed no differences to control cells anymore. Interestingly, Str. uberis in the same concentration did only induce the expression of IL-8 after a 6 h treatment but had no influence on other immunomodulator mRNA expression. Cell culture supernatants of E. coli and S. aureus treated cells for 12 h increased leukocyte chemotaxis in a 96-well MultiScreen<sup>TM</sup>MIC-plate. S. aureus seemed to induce increased chemotaxis after shorter treatments than E. coli. In conclusion, mammary epithelial cells are involved in the different immune response to various mastitis pathogens, and the induction of chemotaxis of leukocytes from blood to milk during mastitis. Therefore, most likely epithelial cells play a role in the differential pattern of immunomediators stimulated by different pathogens.

scholarly journals Lipocalin 2: A New Antimicrobial in Mast Cells

2019 ◽  
Vol 20 (10) ◽  
pp. 2380 ◽  
Author(s):  
Yu-Ling Chang ◽  
Zhenping Wang ◽  
Satomi Igawa ◽  
Jae Eun Choi ◽  
Tyler Werbel ◽  
...  
Keyword(s):  
Mast Cells ◽  
Antimicrobial Peptides ◽  
Antimicrobial Peptide ◽  
Bacterial Growth ◽  
Immune Defense ◽  
Lipocalin 2 ◽  
Vitro Assay ◽  
E Coli ◽  
Sphingosine 1 Phosphate

Mast cells (MCs) play a significant role in the innate immune defense against bacterial infection through the release of cytokines and antimicrobial peptides. However, their antimicrobial function is still only partially described. We therefore hypothesized that MCs express additional antimicrobial peptides. In this study, we used FANTOM 5 transcriptome data to identify for the first time that MCs express lipocalin 2 (LCN2), a known inhibitor of bacterial growth. Using MCs derived from mice which were deficient in LCN2, we showed that this antimicrobial peptide is an important component of the MCs’ antimicrobial activity against Escherichia coli (E. coli). Since sphingosine-1-phosphate receptors (S1PRs) on MCs are known to regulate their function during infections, we hypothesized that S1P could activate LCN2 production in MCs. Using an in vitro assay, we demonstrated that S1P enhances MCs antimicrobial peptide production and increases the capacity of MCs to directly kill S. aureus and E. coli via an LCN2 release. In conclusion, we showed that LCN2 is expressed by MCs and plays a role in their capacity to inhibit bacterial growth.

scholarly journals Characterization of the Bacteriophage-Derived Endolysins PlySs2 and PlySs9 with In Vitro Lytic Activity against Bovine Mastitis Streptococcus uberis

Antibiotics ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 621
Author(s):  
Niels Vander Elst ◽  
Sara B. Linden ◽  
Rob Lavigne ◽  
Evelyne Meyer ◽  
Yves Briers ◽  
...  
Keyword(s):  
Bovine Mastitis ◽  
Binding Activity ◽  
Lytic Activity ◽  
Economic Losses ◽  
Therapeutic Measure ◽  
Cell Wall Binding ◽  
Streptococcus Uberis ◽  
Major Loss

Bovine mastitis, an infection of the cow’s mammary gland, is frequently caused by Streptococcus uberis and causes major economic losses in the dairy industry. The intramammary administration of antibiotics currently remains the predominant preventive and therapeutic measure. These antimicrobial compounds, of which some are considered critical in human health care, are frequently applied as dry therapy resulting in their consistent overuse. Therefore, the use of antibiotics in the dairy sector is being questioned. We here identified two endolysins, i.e., PlySs2 and PlySs9, respectively derived from Streptococcus suis serotype-2 and -9 prophages, with lytic activity against S. uberis in an in vitro setting. Both endolysins gave clear lysis zones in spot-on-plate assays and caused a reduction of the optical density in a turbidity reduction assay. In depth characterization identified PlySs9 as the more potent endolysin over PlySs2 with a lower MIC value and about one additional log of killing. PlySs2 and PlySs9 were challenged to a panel of subclinical and clinical S. uberis milk isolates and were both able to lyse all strains tested. Molecular dissection of these endolysins in catalytic and cell wall binding subdomains resulted in major loss of killing and binding activity, respectively. Taken together, we here propose PlySs2 and PlySs9 as candidate compounds to the current antimicrobial arsenal known against bovine mastitis-causing S. uberis as future add-on or replacement strategy to the currently used intramammary antibiotics.

scholarly journals In Vitro Antimicrobial Activity of an Experimental Dentifrice Based on Ricinus Communis

2014 ◽  
Vol 25 (3) ◽  
pp. 191-196 ◽  
Author(s):  
Vanessa Maria Fagundes Leite ◽  
Juliana Barchelli Pinheiro ◽  
Marina Xavier Pisani ◽  
Evandro Watanabe ◽  
Raphael Freitas de Souza ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Growth Inhibition ◽  
Bacterial Growth ◽  
Ricinus Communis ◽  
Candida Spp ◽  
E Coli ◽  
Reflected Light ◽  
Bacterial Growth Inhibition ◽  
Post Hoc

This study evaluated the antimicrobial activity of a Ricinus communis-based experimental dentifrice for denture hygiene against the following standard strains: Staphylococcus aureus, Escherichia coli, Streptococcus mutans, Enterococcus faecalis, Candida albicans and Candida glabrata. The minimum inhibitory concentration (MIC) assay was performed with R. communis in pure oil at 2.5%. Only E. coli was not inhibited by R. communis, but the MIC (0.0781%) was effective against the other microorganisms. From these results it was determined the R. communis concentrations for experimental dentifrices, 1, 2, 5 and 10%, which were evaluated by the test-well diffusion in agar. The commercial dentifrices Colgate, Trihydral and Corega Brite were tested for comparative purposes. The diameter of the zones of bacterial growth inhibition produced around the wells was measured (in mm) with a rule under reflected light. Data were analyzed statistically by analysis of variance and Tukey's post-hoc test (α=0.05). Neither the commercial nor the experimental dentifrices were effective against E. coli. The experimental dentifrices containing R. communis at 2, 5 and 10% presented action against S. mutans, S. aureaus and E. faecallis. The experimental dentifrices showed no antimicrobial activity against Candida spp. and E. coli in any of the tested concentrations. Trihydral was the most effective. Comparing the experimental dentifrices, the product with 10% R. communis produced the largest zones of bacterial growth inhibition and had similar antimicrobial activity to the commercial dentifrices, except against S. aureus.

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