Effect of bovine lactoferrin on the internalization of coagulase-negative staphylococci into bovine mammary epithelial cells under in-vitro conditions

2009 ◽  
Vol 76 (2) ◽  
pp. 144-151 ◽  
Author(s):  
Paula Hyvönen ◽  
Sari Käyhkö ◽  
Suvi Taponen ◽  
Atte von Wright ◽  
Satu Pyörälä

Coagulase-negative staphylococci (CNS) have emerged as bovine mastitis pathogens in many countries. CNS mastitis is generally mild but can persist in the udder for long periods. Pathogenesis of CNS intramammary infection is not well understood. In the present study, adhesion, invasion and intracellular replication of twenty-two CNS strains isolated from bovine mastitis and the effect of bovine lactoferrin (bLf) on the internalization were studied in vitro in a bovine mammary epithelial (BME) cell model. The CNS strains were ofStaphylococcus chromogenes, Staph. simulans, Staph. epidermidis, Staph. haemolyticusandStaph. cohnii urealyticus; two strains ofStaph. aureuswere used as controls. Seven of the CNS strains originated from persistent and five from transient mastitis infections. The in-vitro susceptibility of the strains to bLf was also investigated. All CNS species examined had an adhesive ability equal to that ofStaph. aureus, but internalization varied among staphylococcal strains. The antagonistic effect of bLf on the adhesion and invasion of CNS strains was weak, but bLf significantly decreased intracellular replication and replication rates of CNS. No correlation between the in-vitro susceptibility of the strain to bLf or internalization among clinical signs of mastitis was established. No difference between the persistent and transient CNS strains in adhesion, invasion or replication rate was recorded. This in-vitro BME cell model can be used to study the virulence potential of mastitis pathogens, although the severity and persistence of eventual infections shall be further investigated in vivo. The role of bLf in intramammary infection caused by CNS may be limited.

2021 ◽  
Vol 88 (1) ◽  
pp. 80-88
Author(s):  
Remo Stürmlin ◽  
Josef J. Gross ◽  
Olga Wellnitz ◽  
Lea A. Wagner ◽  
Camille Monney ◽  
...  

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.


2017 ◽  
Vol 95 (1) ◽  
pp. 69-75 ◽  
Author(s):  
Kei-ichi Shimazaki ◽  
Kazuhiro Kawai

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.


1989 ◽  
Vol 9 (3) ◽  
pp. 179-190 ◽  
Author(s):  
Anders Tranæus ◽  
Olof Heimbürger ◽  
Bengt Lindholm

The analysis of all episodes of peritonitis occurring in a uniformly treated continuous ambulatory peritoneal dial ysis (CAPD) population ( N = 128) at one centre during a six-year period showed the following major findings. The initial white cell count (WCC) of the dialysate was <100 .106/L in 10% of the episodes and showed a predominance of mononuclear cells in 15%. The Gram stain results were consistent with the findings of the culture in 28% of the episodes and influenced the initial therapy in only 7% of the cases. Between 9% and 31% of all episodes would not have been classified as peritonitis if positive culture, a WCC of >100 .106/L in the dialysate, or clinical symptoms had been required for the diagnosis. The proportion of negative dialysate cultures was 2% after the introduction of pre-culture membrane filtration. Tunnel infection as a cause of peritonitis was more frequent in episodes due to Staphylococcus aureus than in episodes due to coagulase-negative staphylococci (p = 0.009). Peritonitis caused by coagulase negative staphylococci were followed by a milder course than other organisms (p = 0.02). Of all episodes initially treated with cephradine only 62% were cured with this antibiotic (or cloxacillin) and 35% were followed by recurrency, protracted course or catheter loss, despite intermediate or full in vitro susceptibility. In only 53% of all episodes no complication was observed. Complications were more frequent in women and diabetics than in men (p = 0.01) and non-diabetics (p = 0.03), and were more common in episodes with clinical symptoms (p = 0.02). Peritonitis resulted in drop-out from CAPD in 6% of all episodes. Hospital care was needed in 68% of all episodes. We conclude that turbidity can be used as the sole criterion for the initial diagnosis of peritonitis, and that a first generation cephalosporin should not be used as a first line antibiotic in the treatment of CAPD peritonitis.


2015 ◽  
Vol 45 (6) ◽  
pp. 1035-1041
Author(s):  
Ananda Paula Kowalski ◽  
Grazieli Maboni ◽  
Julia Pires Espindola ◽  
Ariane Foletto ◽  
Guerino Bandeira Junior ◽  
...  

Records of in vitro susceptibility tests performed between 1992 and 2011 were retrospectively reviewed in order to evaluate the dynamic profiles of possible changes in antimicrobial resistance of Staphylococcus spp. isolated from milk samples of cows with mastitis during two decades. The results of 2,430 isolates tested by disk diffusion technique for susceptibility to oxacillin, penicillin, ampicillin, cephalexin, norfloxacin, tetracycline, sulfazotrim, gentamicin, and neomycin were analysed. Comparisons were performed between the percentages of resistance to antimicrobials and their classes and also between the decades studied. Additionally, the possible tendency or changes in the behaviour of these pathogens against the major drugs used in the last two decades were evaluated using regression analysis. The highest rates of resistance (P<0.0001) were observed for the beta-lactams (34.3%), with exception of cephalexin (6.9%), and for the tetracyclines (28%). Similar resistance rates (7.6% to 15.7%) were observed among the other drugs. Regression analysis showed a reduction in resistance to penicillin and ampicillin throughout the period, whilst for oxacillin and neomycin a decrease in the resistance was observed during the first decade, followed by an increase. A trend towards decreased resistance was found for sulfazotrim, whereas for the other antimicrobials no decrease was observed. The results indicated no trend towards increased resistance for most antimicrobials tested. Nevertheless, it is necessary to monitor the resistance patterns of these pathogens in order to save these drugs as a therapeutic reserve


2018 ◽  
Vol 72 (2) ◽  
pp. 181-184
Author(s):  
Saugata Choudhury ◽  
Lee Kar Mun ◽  
Esme Ng Chu Xuan ◽  
Lee Shin Jia ◽  
Shawn Vasoo ◽  
...  

We compared the in vitro antimicrobial activities of tedizolid and linezolid on the Sensititre broth microdilution system for Gram-positive cocci isolates (n=146) from skin and skin structure infections and bloodstream infections, bronchoalveolar lavage and sputum. These pathogens included 40 methicillin-resistant Staphylococcus aureus, 38 coagulase-negative staphylococci, 20 Enterococcus faecalis and 48 beta-haemolytic Streptococcus spp. Susceptibility was simultaneously determined for 48 vanA vancomycin-resistant enterococci isolates 2013–2016 from rectal swabs (23 E. faecalis and 25 E. faecium, of which 4 were linezolid-non-susceptible). MIC90s for tedizolid were fourfold to eightfold lower than linezolid on the Sensititre and ranged from 0.12 to 0.5 µg/mL for the different pathogen groups. All isolates were susceptible to tedizolid except two vanA E. faecium strains (MICs of 1 and 2 µg/mL, respectively). Categorical and essential agreement for tedizolid were 99.48% and 92%, respectively, between Liofilchem gradient diffusion and Sensititre methods. Overall, the drug exhibited excellent activity against the surveyed Gram-positive pathogens.


2005 ◽  
Vol 171 (4) ◽  
pp. 663-673 ◽  
Author(s):  
Wa Xian ◽  
Kathryn L. Schwertfeger ◽  
Tracy Vargo-Gogola ◽  
Jeffrey M. Rosen

Members of the fibroblast growth factor (FGF) family and the FGF receptors (FGFRs) have been implicated in mediating various aspects of mammary gland development and transformation. To elucidate the molecular mechanisms of FGFR1 action in a context that mimics polarized epithelial cells, we have developed an in vitro three-dimensional HC11 mouse mammary epithelial cell culture model expressing a drug-inducible FGFR1 (iFGFR1). Using this conditional model, iFGFR1 activation in these growth-arrested and polarized mammary acini initially led to reinitiation of cell proliferation, increased survival of luminal cells, and loss of cell polarity, resulting in the disruption of acinar structures characterized by the absence of an empty lumen. iFGFR1 activation also resulted in a gain of invasive properties and the induction of matrix metalloproteinase 3 (MMP-3), causing the cleavage of E-cadherin and increased expression of smooth muscle actin and vimentin. The addition of a pan MMP inhibitor abolished these phenotypes but did not prevent the effects of iFGFR1 on cell proliferation or survival.


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