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Journal of Dairy Science Aug 2020The objective of this study was to evaluate the antibacterial activities of extract derived from moringa leaves. In particular, the effect of moringa extract (Mor) on...
The objective of this study was to evaluate the antibacterial activities of extract derived from moringa leaves. In particular, the effect of moringa extract (Mor) on adhesion and invasion of Escherichia coli O55, Enterococcus faecalis, Staphylococcus simulans, and Serratia liquefaciens was evaluated in bovine mammary epithelial cells (MAC-T). Broth microdilution method, minimum inhibitory concentration and minimum bactericidal concentration assays, adhesion and invasion assays, and real-time PCR were performed. The minimum inhibitory concentration and minimum bactericidal concentration of Mor ranged from 12.5 to 50 mg/mL on 18 out of 27 tested isolates. Treatment of E. coli O55 with Mor (100 and 200 μg/mL) inhibited the adhesion and invasion on MAC-T cells via downregulation of adhesion factors (i.e., papC, f17c-A, and eaeA). Also, when MAC-T cells were pretreated with Mor (200 μg/mL, 12 h) and incubated with E. coli O55, Enterococcus faecalis, Staphylococcus simulans, or Serratia liquefaciens, both E. coli O55 and Enterococcus faecalis showed a significant decrease in adhesion and invasion. Staphylococcus simulans exhibited decreased adhesion and increased invasion. Serratia liquefaciens showed increased adhesion and decreased invasion. In addition, Mor increased mRNA expression of antioxidant enzymes (e.g., heme oxygenase-1, NAD(P)H:quinone oxidoreductase-1, and thioredoxin reductase 1) in MAC-T cells. In conclusion, 12.5 to 50 mg/mL of Mor exhibited antibacterial activity against 18 out of 27 tested isolates. Also, pretreatment of 200 μg/mL of Mor to MAC-T cells modulated adhesion and invasion of E. coli O55 and other mastitis-associated pathogens. Furthermore, Mor increased antioxidant capacities in MAC-T cells, but further in vivo studies are needed.
Topics: Animals; Anti-Bacterial Agents; Bacterial Adhesion; Cattle; Enterococcus faecalis; Epithelial Cells; Escherichia coli; Escherichia coli Infections; Female; Mammary Glands, Animal; Mastitis, Bovine; Moringa; Plant Extracts
PubMed: 32475678
DOI: 10.3168/jds.2019-17774 -
Journal of Dairy Science Jan 2013Mastitis is the most common and detrimental infection of the mammary gland in dairy cows and has a major economic impact on the production of milk and dairy products....
Mastitis is the most common and detrimental infection of the mammary gland in dairy cows and has a major economic impact on the production of milk and dairy products. Bacterial mastitis is caused by several pathogens, and the most frequently isolated bacterial species are coagulase-negative staphylocci (CNS). Although CNS are considered minor mastitis pathogens, the importance of CNS has increased over the years. However, the mechanism and factors involved in CNS intramammary infection are poorly studied and defined. Biofilms have been proposed as an important component in the persistence of CNS intramammary infection. Biofilms are defined as a cluster of bacteria enclosed in a self-produced matrix. The objectives of this study were to investigate the ability of CNS to form biofilms. A total of 255 mastitis-associated CNS isolates were investigated using a standard microtiter plate biofilm assay. The biofilms of some isolates were also observed by using confocal microscopy. The presence of biofilm-associated genes icaA, bap, aap, embP, fbe, and atlE was determined by PCR in the 255 isolates. The 5 dominant species assayed were Staphylococcus chromogenes (n=111), Staphylococcus simulans (n=53), Staphylococcus xylosus (n=25), Staphylococcus haemolyticus (n=15), and Staphylococcus epidermidis (n=13), and these represented 85% of the isolates. The data gathered were analyzed to identify significant links with the data deposited in the Canadian Bovine Mastitis Research Network database. Overall, Staph. xylosus is the species with the strongest ability to form biofilm, and Staph. epidermidis is the species with the lowest ability to form biofilm. Regardless of the species, the presence of icaA, bap, or the combination of multiple genes was associated with a greater ability to form biofilm. A strong relationship between the strength of a biofilm and days in milk was also noted, and CNS isolated later in the lactation cycle appeared to have a greater ability to form biofilm than those isolated earlier in the lactation cycle. In conclusion, Staph. xylosus is the species with the strongest biofilm formation ability. Furthermore, days in milk and gene combinations are predicted to be the variables with the strongest effect on biofilm formation by CNS.
Topics: Animals; Biofilms; Canada; Cattle; DNA, Bacterial; Female; Mastitis, Bovine; Microscopy, Confocal; Milk; Staphylococcal Infections; Staphylococcus; Staphylococcus epidermidis
PubMed: 23141829
DOI: 10.3168/jds.2012-5795 -
BMC Microbiology Jun 2012Lysostaphin and the catalytic domain of LytM cleave pentaglycine crossbridges of Staphylococcus aureus peptidoglycan. The bacteriocin lysostaphin is secreted by...
BACKGROUND
Lysostaphin and the catalytic domain of LytM cleave pentaglycine crossbridges of Staphylococcus aureus peptidoglycan. The bacteriocin lysostaphin is secreted by Staphylococcus simulans biovar staphylolyticus and directed against the cell walls of competing S. aureus. LytM is produced by S. aureus as a latent autolysin and can be activated in vitro by the removal of an N-terminal domain and occluding region.
RESULTS
We compared the efficacies of the lysostaphin and LytM catalytic domains using a newly developed model of chronic S. aureus infected eczema. Lysostaphin was effective, like in other models. In contrast, LytM was not significantly better than control. The different treatment outcomes could be correlated with in vitro properties of the proteins, including proteolytic stability, affinity to cell wall components other than peptidoglycan, and sensitivity to the ionic milieu.
CONCLUSIONS
Although lysostaphin and LytM cleave the same peptide bond in the peptidoglycan, the two enzymes have very different environmental requirements what is reflected in their contrasting performance in mouse eczema model.
Topics: Animals; Anti-Bacterial Agents; Bacterial Proteins; Biological Products; Catalytic Domain; Disease Models, Animal; Eczema; Endopeptidases; Lysostaphin; Mice; Staphylococcal Skin Infections; Staphylococcus aureus; Treatment Outcome
PubMed: 22672475
DOI: 10.1186/1471-2180-12-97 -
3 Biotech Jun 2016The diversity of some of the culturable microorganisms associated with marine flora and fauna collected off Vizhinjam and Mulloor coast of South India was evaluated and...
The diversity of some of the culturable microorganisms associated with marine flora and fauna collected off Vizhinjam and Mulloor coast of South India was evaluated and their bioactive production potential determined. From a total of 24 bacteria, 4 actinomycetes and 8 fungi isolated from diverse marine sources, five bacterial species-BLM3, BSP2, BCS1, BCS4 and BMA6 showed inhibitory activity against at least one of the tested pathogens viz., Klebsiella pneumonia KU1, Pseudomonas aeruginosa VL3, Salmonella enterica typhimurium MTCC 98, Escherichia coli MTCC 40, Micrococcus luteus MTCC 105, Staphylococcus simulans MTCC 3610, Proteus vulgaris MTCC 426, Vibrio fluvialis, Vibrio sp. P3a and Vibrio sp. P3b. The isolated actinomycetes and fungi did not produce significant inhibition zones against the tested pathogens; however, the macroalgal isolated actinomycetes strain AMA1 produced reddish pigment in Starch Casein medium which remained stable till the stationary phase of growth. The marine sediment isolate BCS4, identified as Bacillus sp. showed wide spectrum of activity against the tested Gram positive bacteria, S. simulans MTCC 3610 and Gram negative bacteria, Proteus vulgaris with zone of inhibitions of 25 and 11 mm respectively. Better extraction of the bioactive compound was obtained with ethyl acetate when compared with methanol, benzene and hexane and TLC analysis revealed the presence of an active compound. The 16SrRNA sequencing confirmed the potent strain belong to Bacillus sp. and hence designated Bacillus sp. BCS4.
PubMed: 28330077
DOI: 10.1007/s13205-015-0318-1 -
Scientific Reports Jul 2017Herein we describe an association between activation of inflammatory pathways following transient hypoxia and the appearance of the multidrug resistant bacteria...
Herein we describe an association between activation of inflammatory pathways following transient hypoxia and the appearance of the multidrug resistant bacteria Staphylococcus simulans in the fetal brain. Reduction of maternal arterial oxygen tension by 50% over 30 min resulted in a subseiuent significant over-expression of genes associated with immune responses 24 h later in the fetal brain. The activated genes were consistent with stimulation by bacterial lipopolysaccharide; an influx of macrophages and appearance of live bacteria were found in these fetal brains. S. simulans was the predominant bacterial species in fetal brain after hypoxia, but was found in placenta of all animals. Strains of S. simulans from the placenta and fetal brain were equally highly resistant to multiple antibiotics including methicillin and had identical genome sequences. These results suggest that bacteria from the placenta invade the fetal brain after maternal hypoxia.
Topics: Animals; Brain; Drug Resistance, Multiple, Bacterial; Female; Fetal Hypoxia; Gene Expression Regulation, Developmental; Macrophages; Microglia; Placenta; Pregnancy; Sheep; Staphylococcus
PubMed: 28743956
DOI: 10.1038/s41598-017-06789-6 -
Applied and Environmental Microbiology Aug 2003Detection of six species of lactic acid bacteria and six species of gram-positive catalase-positive cocci from low-acid fermented sausages (fuets and chorizos) was...
Detection of six species of lactic acid bacteria and six species of gram-positive catalase-positive cocci from low-acid fermented sausages (fuets and chorizos) was assessed by species-specific PCR. Without enrichment, Lactobacillus sakei and Lactobacillus curvatus were detected in 11.8% of the samples, and Lactobacillus plantarum and Staphylococcus xylosus were detected in 17.6%. Enriched samples allowed the detection of L. sakei and S. xylosus in all of the samples (100%) and of Enterococcus faecium in 11.8% of the sausages. The percentages of L. curvatus, L. plantarum, Staphylococcus carnosus, and Staphylococcus epidermidis varied depending on the sausage type. L. curvatus was detected in 80% of fuets and in 57% of chorizos. L. plantarum was found in 50% of fuets and 100% of chorizos. S. epidermidis was detected in only 11.8% of fuets, and S. carnosus was detected in only 5.9% of chorizos. Lactococcus lactis, Staphylococcus warneri, and Staphylococcus simulans were not detected in any sausage type. From a microbiological point of view, 70.6% of the samples could be considered of high quality, as they had low counts of Enterobacteriaceae and did not contain any of the food-borne pathogens assayed.
Topics: Base Sequence; Colony Count, Microbial; Lactobacillus; Meat Products; Molecular Sequence Data; Polymerase Chain Reaction; Species Specificity; Staphylococcus
PubMed: 12902246
DOI: 10.1128/AEM.69.8.4583-4594.2003 -
Applied and Environmental Microbiology Apr 1995Staphylococcus simulans biovar staphylolyticus produces an extracellular glycylglycine endopeptidase (lysostaphin) that lyses other staphylococci by hydrolyzing the...
Staphylococcus simulans biovar staphylolyticus produces an extracellular glycylglycine endopeptidase (lysostaphin) that lyses other staphylococci by hydrolyzing the cross bridges in their cell wall peptidoglycans. The genes for endopeptidase (end) and endopeptidase resistance (epr) reside on plasmid pACK1. An 8.4-kb fragment containing end was cloned into shuttle vector pL150 and was then introduced into Staphylococcus aureus RN4220. The recombinant S. aureus cells produced endopeptidase and were resistant to lysis by the enzyme, which indicated that the cloned fragment also contained epr. Treatments to remove accessory wall polymers (proteins, teichoic acids, and lipoteichoic acids) did not change the endopeptidase sensitivity of walls from strains of S. simulans biovar staphylolyticus or of S. aureus with and without epr. Immunological analyses of various wall fractions showed that there were epitopes associated with endopeptidase resistance and that these epitopes were found only on the peptidoglycans of epr+ strains of both species. Treatment of purified peptidoglycans with endopeptidase confirmed that resistance or susceptibility of both species was a property of the peptidoglycan itself. A comparison of the chemical compositions of these peptidoglycans revealed that cross bridges in the epr+ cells contained more serine and fewer glycine residues than those of cells without epr. The presence of the 8.4-kb fragment from pACK1 also increased the susceptibility of both species to methicillin.
Topics: Cell Wall; Cloning, Molecular; Drug Resistance, Microbial; Genes, Bacterial; Glycine; Lysostaphin; Peptidoglycan; Serine; Staphylococcus; Staphylococcus aureus
PubMed: 7747966
DOI: 10.1128/aem.61.4.1475-1479.1995 -
Foods (Basel, Switzerland) Sep 2020Seventeen staphylococci isolated from 54 Slovak local lump cheeses made from ewes' milk were taxonomically allotted to five species and three clusters/groups involving...
Seventeen staphylococci isolated from 54 Slovak local lump cheeses made from ewes' milk were taxonomically allotted to five species and three clusters/groups involving the following species: (5 strains), (3 strains), (one strain) (5 strains) and (3 strains). Five different species were determined. The aim of the study follows two lines: basic research in connection with staphylococci, and further possible application of the bacteriocins. Identified staphylococci were mostly susceptible to antibiotics (10 out of 14 antibiotics). Strains showed γ-hemolysis (meaning they did not form hemolysis) except for SAOS1/1 strain, which formed β-hemolysis. SAOS1/1 strain was also DNase positive as did SAOS5/2 and SAOS51/3. The other staphylococci were DNase negative. SAOS1/1 and SAOS51/3 showed biofilm formation on Congo red agar. However, using quantitative plate assay, 12 strains out of 17 showed low-grade biofilm formation (0.1 ≤ A < 1), while five strains did not form biofilm (A < 0.1). The growth of all strains, including those strains resistant to enterocins, was inhibited by nisin and gallidermin, with high inhibition activity resulting in the inhibition zone in size from 1600 up to 102,400 AU/mL (arbitrary unit per milliliter). This study contributes to microbiota colonization associated with raw ewe's milk lump cheeses; it also indicates bacteriocin treatment benefit, which can be used in prevention and/or elimination of staphylococci.
PubMed: 32971750
DOI: 10.3390/foods9091335 -
Journal of Dairy Science Feb 1991The utility of trehalose-mannitol broth and arabinose-cellobiose broth for identification of Staphylococcus epidermidis and novobiocin-resistant staphylococci was...
The utility of trehalose-mannitol broth and arabinose-cellobiose broth for identification of Staphylococcus epidermidis and novobiocin-resistant staphylococci was determined using 236 coagulase-negative staphylococci isolated from bovine mammary glands. None of the 49 S. epidermidis strains was positive in trehalose-mannitol broth; whereas, all strains of Staphylococcus hyicus, Staphylococcus chromogenes, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus warneri, and Staphylococcus simulans were positive. Of the novobiocin-resistant staphylococcal species, only Staphylococcus saprophyticus was negative in arabinose-cellobiose broth. Except for one strain of Staphylococcus sciuri and one strain of Staphylococcus kloosii, all remaining strains of novobiocin-resistant staphylococcal species were positive in arabinose-cellobiose broth. Results indicate that trehalose-mannitol broth is an acceptable method for identification of S. epidermidis isolated from bovine mammary glands. Furthermore, arabinose-cellobiose broth is a useful method of screening for novobiocin-resistant staphylococci.
Topics: Animals; Cattle; Coagulase; Culture Media; Female; Mammary Glands, Animal; Novobiocin; Staphylococcus; Staphylococcus epidermidis
PubMed: 2045549
DOI: 10.3168/jds.S0022-0302(91)78186-1 -
Antimicrobial Agents and Chemotherapy Jan 1990A penicillin-binding protein of molecular weight 76,000 inducible by beta-lactams was detected in methicillin-resistant Staphylococcus haemolyticus and Staphylococcus...
A penicillin-binding protein of molecular weight 76,000 inducible by beta-lactams was detected in methicillin-resistant Staphylococcus haemolyticus and Staphylococcus simulans. DNA from these strains hybridized to the mecA gene from Staphylococcus aureus; however, the chromosomal HindIII fragments containing the mecA genes were 3.4 kilobases in S. haemolyticus and 4.3 kilobases in S. simulans.
Topics: Bacterial Proteins; Blotting, Southern; Carrier Proteins; Culture Media; DNA, Bacterial; Deoxyribonuclease HindIII; Electrophoresis, Polyacrylamide Gel; Gene Expression Regulation, Bacterial; Genes, Bacterial; Hexosyltransferases; Methicillin; Muramoylpentapeptide Carboxypeptidase; Nucleic Acid Hybridization; Penicillin Resistance; Penicillin-Binding Proteins; Peptidyl Transferases; Restriction Mapping; Staining and Labeling; Staphylococcus; Staphylococcus aureus
PubMed: 1691614
DOI: 10.1128/AAC.34.1.170