-
Scientific Reports Apr 2019Staphylococcus simulans lysostaphin cleaves pentaglycine cross-bridges between stem peptides in the peptidoglycan of susceptible staphylococci, including S. aureus. This...
Staphylococcus simulans lysostaphin cleaves pentaglycine cross-bridges between stem peptides in the peptidoglycan of susceptible staphylococci, including S. aureus. This enzyme consists of an N-terminal catalytic domain and a cell wall binding domain (SH3b), which anchors the protein to peptidoglycan. Although structures of SH3bs from lysostaphin are available, the binding modes of peptidoglycan to these domains are still unclear. We have solved the crystal structure of the lysostaphin SH3b domain in complex with a pentaglycine peptide representing the peptidoglycan cross-bridge. The structure identifies a groove between β1 and β2 strands as the pentaglycine binding site. The structure suggests that pentaglycine specificity of the SH3b arises partially directly by steric exclusion of Cβ atoms in the ligand and partially indirectly due to the selection of main chain conformations that are easily accessible for glycine, but not other amino acid residues. We have revealed further interactions of SH3b with the stem peptides with the support of bioinformatics tools. Based on the structural data we have attempted engineering of the domain specificity and have investigated the relevance of the introduced substitutions on the domain binding and specificity, also in the contexts of the mature lysostaphin and of its bacteriolytic activity.
Topics: Amino Acid Sequence; Computational Biology; Computer Simulation; Escherichia coli; Lysostaphin; Models, Molecular; Peptidoglycan; Protein Binding; Protein Conformation; Protein Domains; Protein Engineering; Staphylococcus
PubMed: 30979923
DOI: 10.1038/s41598-019-42435-z -
Cureus Nov 2022A toddler girl presented to our hospital with a fever that lasted for five days. She had no prior history of urinary tract infections or contact with farm animals....
A toddler girl presented to our hospital with a fever that lasted for five days. She had no prior history of urinary tract infections or contact with farm animals. Investigations revealed a diagnosis of acute focal bacterial nephritis (AFBN), and we initiated antimicrobial therapy with ampicillin and cefmetazole. On day five, methicillin-resistant coagulase-negative were detected in her urine culture, and we changed the antibiotics to vancomycin. Antibiotic therapy was continued for 21 days, with no recurrence of fever. Finally, the bacteria were identified as , which is a common farm animal pathogen. Clinicians should be aware of the possibility of AFBN caused by , even if the patient has no prior history of close contact with farm animals. If a rare organism is detected in urine culture during AFBN treatment, the patient should be treated with appropriate antibiotics for the pathogen.
PubMed: 36505107
DOI: 10.7759/cureus.31241 -
Meat Science Feb 2020This study focused on sarcoplasmic and myofibrillar protein degradation and the formation of peptides with antioxidant activity by mixed starters (Lactobacillus...
This study focused on sarcoplasmic and myofibrillar protein degradation and the formation of peptides with antioxidant activity by mixed starters (Lactobacillus plantarum CD101 and Staphylococcus simulans NJ201). Gel electrophoresis indicated that the mixed starters can hydrolyze both sarcoplasmic and myofibrillar proteins, and the concentration of peptides increased (P < .05). Compared with the control group, using mixed starters led to a significant increase (P < .05) in the DPPH radical scavenging activity, Fe chelating activity, and ABTS radical scavenging activity of sarcoplasmic proteins, but demonstrated no significant difference in myofibrillar proteins. Two hydrophobic fractions (C2, C5) separated by RP-HPLC in the inoculation groups with sarcoplasmic proteins showed high DPPH radical scavenging activity (66.60%, 60.50%). Eighteen peptides were identified by LC-MS/MS, which mainly arose from triosephosphate isomerase, creatine kinase M-type, and glyceraldehyde-3-phosphate dehydrogenase. Hydrophobic amino acids accounted for a large proportion. Our results indicate that mixed starters affect proteolytic characterization and contribute to the formation of peptides with antioxidant capacity in sarcoplasmic proteins.
Topics: Animals; Antioxidants; Fermentation; Lactobacillus plantarum; Meat Products; Muscle Proteins; Myofibrils; Peptides; Proteolysis; Staphylococcus; Swine
PubMed: 31669862
DOI: 10.1016/j.meatsci.2019.107958 -
Animal Biotechnology Dec 2023This study aimed to investigate the presence of and genes in coagulase negative (CNS) species isolated from bovine mastitis in smallholder dairy farms by using PCR. A...
This study aimed to investigate the presence of and genes in coagulase negative (CNS) species isolated from bovine mastitis in smallholder dairy farms by using PCR. A total of 602 mammary quarter milk samples belong to 170 cows with mastitis were used. Identification of species was achieved by using the commercial Gram-positive identification kit and a total of 52 (8.6%) CNS species were isolated. The most frequently isolated species was (n = 15, 28.8%), followed by (n = 12, 23.1%), (n = 8, 15.4%), (n = 5, 9.6%), (n = 4, 7.7%), (n = 4, 7.7%), (n = 2, 3.8%) and (n = 2, 3.8%). The gene positivity was found in the 13 (25%) of strains. Of the strains carrying gene, eight also harbored the gene. A total of gene positivity was found as 30.8% (n = 16) in 52 CNS species. In conclusion, the present study showed that CNS isolated from cows with mastitis may be reservoir of and genes. To our knowledge, this is the first study showing the presence of and genes in CNS species isolated from bovine with mastitis in the smallholder dairy farms in Turkey.
Topics: Female; Animals; Cattle; Coagulase; Mastitis, Bovine; Staphylococcal Infections; Prevalence; Farms; Turkey; Staphylococcus; Milk; Cattle Diseases
PubMed: 35792781
DOI: 10.1080/10495398.2022.2094802 -
Journal of Dairy Science Jun 2018The purpose of this study was to investigate non-aureus Staphylococcus spp. intramammary infections (IMI) in periparturient heifers and determine the relationship of...
The purpose of this study was to investigate non-aureus Staphylococcus spp. intramammary infections (IMI) in periparturient heifers and determine the relationship of precalving body site isolation with precalving IMI and postcalving IMI using molecular speciation and strain-typing methods. Primiparous heifers were enrolled at approximately 14 d before expected calving date. Precalving mammary quarter secretions and body site swabbing samples (teat skin, inguinal skin, muzzle, and perineum) were collected. Postcalving, mammary quarter milk samples were collected for culture and somatic cell counting. Precalving body site samples were cultured, and up to 10 staphylococcal colonies were saved for characterization. Staphylococcal isolates were speciated using matrix-assisted laser/desorption ionization time-of-flight mass spectrometry or sequencing of rpoB or tuf. Pulsed-field gel electrophoresis was used to strain type a subset of isolates. Overall, Staphylococcus chromogenes, Staphylococcus agnetis, and Staphylococcus simulans were the most common species identified in precalving mammary secretions, whereas S. chromogenes, Staphylococcus xylosus, and S. agnetis were the most common species found in postcalving milk samples. The most common species identified from body site samples were S. chromogenes, S. xylosus, and Staphylococcus haemolyticus. Mammary quarters that had a precalving mammary secretion that was culture positive for S. agnetis, S. chromogenes, or Staphylococcus devriesei had increased odds of having an IMI with the same species postcalving. A S. chromogenes IMI postcalving was associated with higher somatic cell count when compared with postcalving culture-negative quarters. Among heifers identified with a non-aureus Staphylococcus spp. IMI either precalving or postcalving, heifers that had S. agnetis or S. chromogenes isolated from their teat skin had increased odds of having the same species found in their precalving mammary secretions, and heifers with S. chromogenes, S. simulans, and S. xylosus isolated from their teat skin precalving were at increased odds of having an IMI with the same species postcalving. Overall, 44% of all heifers with a S. chromogenes IMI around the time of parturition had the same strain isolated from a body site. Based on pulsed-field gel electrophoresis, a high level of strain diversity was found.
Topics: Animals; Cattle; Cell Count; Electrophoresis, Gel, Pulsed-Field; Female; Mammary Glands, Animal; Mastitis, Bovine; Milk; Staphylococcal Infections; Staphylococcus
PubMed: 29525303
DOI: 10.3168/jds.2017-13910 -
Probiotics and Antimicrobial Proteins Aug 2023Bacteriocins are antimicrobial peptides produced by bacteria. This study aimed to in silico analyze the presence of bacteriocin gene clusters (BGCs) among the genomes of...
Bacteriocins are antimicrobial peptides produced by bacteria. This study aimed to in silico analyze the presence of bacteriocin gene clusters (BGCs) among the genomes of 22 commensal Staphylococcus isolates from different origins (environment/human/food/pet/wild animals) previously identified as bacteriocin producers. The resistome and plasmidome were studied in all isolates. Five types of BGC were detected in 18 genomes of the 22 bacteriocin-producing staphylococci included in this study: class I (Lanthipeptides), class II, circular bacteriocins, the non-ribosomal-peptide lugdunin and the thiopeptide micrococcin P1 (MP1). A high frequency of lanthipeptides was detected in this collection: BGC variants of BSA, bacCH91, and epilancin15X were identified in two Staphylococcus aureus and one Staphylococcus warneri isolates from food and wild animals. Moreover, two potentially new lanthipeptide-like BGCs with no identity to database entries were found in Staphylococcus epidermidis and Staphylococcus simulans from food and wild animal, respectively. Interestingly, four isolates (one S. aureus and one Staphylococcus hominis, environmental origin; two Staphylococcus sciuri, food) carried the MP1 BGC with differences to those previously described. On the other hand, seven of the 22 genomes (~32%) lacked known genes related with antibiotic or disinfectant-acquired resistance mechanisms. Moreover, the potential carriage of plasmids was evaluated, and several Rep-proteins were identified (~73% of strains). In conclusion, a wide variety of BGCs has been observed among the 22 genomes, and an interesting relationship between related Staphylococcus species and the type of bacteriocin has been revealed. Therefore, bacteriocin-producing Staphylococcus and especially coagulase-negative staphylococci (CoNS) can be considered good candidates as a source of novel bacteriocins.
PubMed: 37632676
DOI: 10.1007/s12602-023-10119-w -
Foodborne Pathogens and Disease Jul 2015Bacteria harboring cfr, a multidrug resistance gene, have high prevalence in livestock in China and might be transmitted to humans through direct contact or via...
Distribution of the Multidrug Resistance Gene cfr in Staphylococcus Isolates from Pigs, Workers, and the Environment of a Hog Market and a Slaughterhouse in Guangzhou, China.
Bacteria harboring cfr, a multidrug resistance gene, have high prevalence in livestock in China and might be transmitted to humans through direct contact or via contaminated food products. To better understand the epidemiology of cfr producers in the food chain, the prevalence and genetic analysis of Staphylococcus isolates recovered from pigs, workers, and meat-handling facilities (a slaughterhouse and a hog market in Guangzhou, China) were examined. Twenty (4.5%) cfr-positive Staphylococcus isolates (18 Staphylococcus simulans, 1 S. cohnii, and 1 S. aureus) were derived from pigs (16/312), the environment (2/52), and workers (2/80). SmaI pulsed-field gel electrophoresis of 26 staphylococcal strains (22 S. simulans and 4 S. cohnii), including previously reported cfr-carrying staphylococci of animal food origin, exhibited 19 major pulsed-field gel electrophoresis patterns (A-S). Clonal spread of cfr-carrying staphylococci among pigs, workers, and meat products was detected. The genetic contexts of cfr in plasmids (pHNKF3, pHNZT2, and pHNCR35) obtained from S. simulans of swine or human origin were similar to that of Staphylococcus species isolated from human clinics and animal-derived food. The cfr-carrying S. aureus strain isolated from floor swabs of the hog market was spa-type t889 and belonged to the ST9 clonal lineage. In summary, both clonal spread and horizontal transmission via mobile elements contributed to cfr dissemination among staphylococcal isolates obtained from different sources. To monitor potential outbreaks of cfr-positive bacteria, continued surveillance of this gene in animals at slaughter and in animal-derived food is warranted.
Topics: Abattoirs; Animals; Anti-Bacterial Agents; Bacterial Proteins; China; DNA, Bacterial; Drug Resistance, Multiple, Bacterial; Electrophoresis, Gel, Pulsed-Field; Food Contamination; Food Handling; Food Microbiology; Genes, MDR; Humans; Meat Products; Microbial Sensitivity Tests; Multilocus Sequence Typing; Plasmids; Staphylococcus; Swine
PubMed: 25974310
DOI: 10.1089/fpd.2014.1891 -
Food Microbiology Dec 2018Coagulase-negative staphylococci (CNS) contribute to the product quality of fermented meats. In spontaneously fermented meats, CNS communities are variable and difficult...
Coagulase-negative staphylococci (CNS) contribute to the product quality of fermented meats. In spontaneously fermented meats, CNS communities are variable and difficult to predict, as their compositions depend on a superposed combination of different processing factors. To partially disentangle this superposition, a meat model system was used to study the influence of temperature and pH on the CNS community dynamics. Therefore, cured pork mince was prepared that was divided into three batches of different initial acidity levels, namely pH 5.7, pH 5.5, and pH 5.3. These three batches were incubated at three different temperatures, namely 23 °C, 30 °C, and 37 °C. Hence, the experimental set-up resulted in nine combinations of different temperature and initial pH values. Samples were analysed after 3 and 14 days to monitor pH, colony counts, and species diversity of the CNS communities, based on mannitol-salt-phenol-red agar (MSA) medium. At conditions of mild acidity (pH 5.7) and low temperature (23 °C), as often encountered during artisan-type meat fermentations, a co-prevalence of Staphylococcus xylosus, Staphylococcus equorum, and Staphylococcus saprophyticus occurred. At the same initial pH but higher incubation temperatures (30 °C and 37 °C), Staphylococcus lugdunensis became the prevailing CNS species, besides S. saprophyticus (30 °C) and the coagulase-positive species Staphylococcus aureus (37 °C). When the initial pH was set at 5.5, S. saprophyticus was the prevailing CNS species at both 23 °C and 30 °C, but it was replaced by Staphylococcus epidermidis and Staphylococcus simulans at 37 °C after 3 and 14 days, respectively. At the most acidic conditions (pH 5.3), CNS counts declined and many of the MSA isolates were of non-staphylococcal nature. Among others, Staphylococcus carnosus (23 °C), Staphylococcus warneri (30 °C), and S. epidermidis (37 °C) were found. Overall, the results of the present study indicated that the processing factors temperature and pH had a clear impact on the shaping of staphylococcal communities during meat fermentation.
Topics: Animals; Bioreactors; Coagulase; Fermentation; Fermented Foods; Food Microbiology; Hydrogen-Ion Concentration; Meat; Meat Products; Microbial Consortia; Models, Biological; Red Meat; Staphylococcus; Swine; Temperature
PubMed: 30166139
DOI: 10.1016/j.fm.2018.05.006 -
Journal of Global Antimicrobial... Mar 2021To evaluate the in vitro activity of dalbavancin compared with vancomycin, daptomycin and other agents against a large collection of coagulase-negative staphylococci...
OBJECTIVES
To evaluate the in vitro activity of dalbavancin compared with vancomycin, daptomycin and other agents against a large collection of coagulase-negative staphylococci (CoNS) isolates.
METHODS
A total of 5088 CoNS causing clinically significant infection were consecutively collected from 122 medical centres in the USA and Europe over 6 years (2014-2019). Isolates were tested for susceptibility by the reference broth microdilution method. Species identification was confirmed by MALDI-TOF. Most isolates were from bloodstream infections (BSIs) (53.5%) or skin/skin structure infections (28.5%).
RESULTS
Staphylococcus epidermidis was the most common species overall (54.6%) and for BSI (61.3%). The second most common species were Staphylococcus lugdunensis overall (12.3%) and Staphylococcus hominis for BSI (14.7%). Dalbavancin (MIC, 0.03/0.06 mg/L) inhibited >99.9% of CoNS isolates at ≤0.25 mg/L (susceptible breakpoint for Staphylococcus aureus per CLSI). All species were inhibited at ≤0.25 mg/L dalbavancin, except some S. epidermidis (>99.9%) and Staphylococcus warneri (98.9%) isolates. Staphylococcus capitis and Staphylococcus simulans exhibited the lowest dalbavancin MIC values (0.015/0.03 mg/L) and Staphylococcus haemolyticus and Staphylococcus saprophyticus the highest (MIC, 0.06/0.12 mg/L); 47.8% of S. epidermidis and 34.7% of S. haemolyticus exhibited decreased susceptibility to vancomycin (MIC ≥ 2 mg/L) and 23.2% of S. capitis and 28.4% of S. warneri showed decreased susceptibility to daptomycin (MIC ≥ 1 mg/L).
CONCLUSION
Antimicrobial susceptibility varied widely among CoNS species. Dalbavancin inhibited >99.9% and 99.1% of isolates at the US-FDA and EUCAST breakpoints, respectively. Clinical studies of dalbavancin for treatment of CoNS infections should be considered based on these in vitro data.
Topics: Anti-Bacterial Agents; Coagulase; Europe; Microbial Sensitivity Tests; Staphylococcus; Teicoplanin
PubMed: 33285311
DOI: 10.1016/j.jgar.2020.11.020 -
The Journal of Dairy Research May 2019The objectives of the research described here were to describe the persistence of intramammary infections (IMI) caused by coagulase negative staphylococci (CNS) in goats...
The objectives of the research described here were to describe the persistence of intramammary infections (IMI) caused by coagulase negative staphylococci (CNS) in goats using strain-typing, and to evaluate the relationship between species-specific CNS IMI and somatic cell score (SCS) at the udder-half level. Udder-half milk samples were collected from all 909 lactating goats (1817 halves; 1 blind half) in a single herd. Milk samples were cultured on Columbia blood agar, and 220 goats with at least one half yielding a single colony type CNS were enrolled for two additional half-level samplings at approximately 1-month intervals. Isolates were identified to the species level by matrix-assisted laser desorption-ionisation time-of-flight mass spectrometry or PCR amplification and partial sequencing of tuf or rpoB. An IMI was defined as persistent when ≥1 follow-up sample yielded the same species and strain as on Day 0 based on pulsed-field gel electrophoresis. A generalised mixed linear model was used to evaluate the odds of persistence as a function of CNS species. A mixed linear model was used to evaluate the relationship between IMI status on a given day and SCS. Among 192 IMI, 69.8% were persistent based on species and strain-type. Staphylococcus simulans IMI had higher odds of persistence than Staphylococcus arlettae IMI. In primiparous goats, Staphylococcus epidermidis IMI was associated with higher SCS than S. arlettae, Staphylococcus xylosus and 'other CNS' IMI. The differences detected in the present study between CNS species, with regard to persistence of IMI and association with SCS, highlight the need to study CNS at the species and strain level to understand the pathogenicity and epidemiology of CNS in goats.
Topics: Animals; Cross-Sectional Studies; Female; Goat Diseases; Goats; Mastitis; Milk; Staphylococcal Infections; Staphylococcus
PubMed: 31138341
DOI: 10.1017/S0022029919000311