ABSTRACT
Infections caused by Staphylococcus aureus pose a significant global public problem. Therefore, new antibiotics and therapeutic strategies are needed to combat this pathogen. This investigation delves into the effects of iclaprim, a newly discovered inhibitor of folic acid synthesis, on S. aureus virulence. The phenotypic and genotypic effects of iclaprim were thoroughly examined in relation to virulence factors, biofilm formation, and dispersal, as well as partial virulence-encoding genes associated with exoproteins, adherence, and regulation in S. aureus MW2, N315, and ATCC 25923. Then, the in vivo effectiveness of iclaprim on S. aureus pathogenicity was explored by a Galleria mellonella larvae infection model. The use of iclaprim at sub-inhibitory concentrations (sub-MICs) resulted in a reduction of α-hemolysin (Hla) production and a differential effect on the activity of coagulase in S. aureus strains. The results of biofilm formation and eradication assay showed that iclaprim was highly effective in depolymerizing the mature biofilm of S. aureus strains at concentrations of 1 MIC or greater, however, inhibited the biofilm-forming ability of only strains N315 and ATCC 25923 at sub-MICs. Interestingly, treatment of strains with sub-MICs of iclaprim resulted in significant stimulation or suppression of most virulence-encoding genes expression. Iclaprim did not affect the production of δ-hemolysin or staphylococcal protein A (SpA), nor did it impact the total activity of proteases, nucleases, and lipases. In vivo testing showed that sub-MICs of iclaprim significantly improves infected larvae survival. The present study offered valuable insights towards a better understating of the influence of iclaprim on different strains of S. aureus. The findings suggest that iclaprim may have potential as an anti-virulence and antibiofilm agent, thus potentially mitigating the pathogenicity of S. aureus and improving clinical outcomes associated with infections caused by this pathogen. KEY POINTS: ⢠Iclaprim effectively inhibits α-hemolysin production and biofilm formation in a strain-dependent manner and was an excellent depolymerizing agent of mature biofilm ⢠Iclaprim affected the mRNA expression of virulence-encoding genes associated with exoproteins, adherence, and regulation ⢠In vivo study in G. mellonella larvae challenged with S. aureus exhibited that iclaprim improves larvae survival.
Subject(s)
Anti-Bacterial Agents , Biofilms , Larva , Microbial Sensitivity Tests , Staphylococcal Infections , Staphylococcus aureus , Virulence Factors , Staphylococcus aureus/drug effects , Staphylococcus aureus/pathogenicity , Staphylococcus aureus/genetics , Biofilms/drug effects , Animals , Virulence Factors/genetics , Anti-Bacterial Agents/pharmacology , Virulence/drug effects , Staphylococcal Infections/microbiology , Staphylococcal Infections/drug therapy , Larva/microbiology , Moths/microbiology , Hemolysin Proteins/genetics , Folic Acid/pharmacology , Folic Acid/biosynthesis , Folic Acid Antagonists/pharmacology , Coagulase/metabolism , Disease Models, Animal , PyrimidinesABSTRACT
BACKGROUND: Coagulase-negative Staphylococcus species are an emerging cause of intramammary infection, posing a significant economic and public health threat. The aim of this study was to assess the occurrence of coagulase-negative Staphylococcus species in bovine milk and dairy farms in Northwestern Ethiopia and to provide information about their antibiotic susceptibility and virulence gene profiles. METHODS: The cross-sectional study was conducted from February to August 2022. Coagulase-negative Staphylococcus species were isolated from 290 milk samples. Species isolation and identification were performed by plate culturing and biochemical tests and the antimicrobial susceptibility pattern of each isolate was determined by the Kirby-Bauer disc diffusion test. The single-plex PCR was used to detect the presence of virulent genes. The STATA software version 16 was used for data analysis. The prevalence, proportion of antimicrobial resistance and the number of virulent genes detected from coagulase-negative Staphylococcus species were analyzed using descriptive statistics. RESULTS: Coagulase-negative Staphylococcus species were isolated in 28.6%, (95% CI: 23.5-34.2) of the samples. Of these, the S. epidermidis, S. sciuri, S. warneri, S. haemolyticus, S. simulans, S. chromogens, S. cohnii, and S. captis species were isolated at the rates of 11, 5.2, 3.4, 3.1, 3.1, 1, 1, and 0.7% respectively. All the isolates showed a high percentage (100%) of resistance to Amoxicillin, Ampicillin, and Cefotetan and 37.5% of resistance to Oxacillin. The majority (54.2%) of coagulase-negative isolates also showed multidrug resistance. Coagulase-negative Staphylococcus species carried the icaD, pvl, mecA, hlb, sec, and hla virulent genes at the rates of 26.5%, 22.1%, 21.7%, 9.6%, 9.6% and 8.4% respectively. CONCLUSION: The present study revealed that the majority of the isolates (54.2%) were found multidrug-resistant and carriage of one or more virulent and enterotoxin genes responsible for intramammary and food poisoning infections. Thus, urgent disease control and prevention measures are warranted to reduce the deleterious impact of coagulase-negative species. To the best of our knowledge, this is the first study in Ethiopia to detect coagulase-negative Staphylococcus species with their associated virulent and food poisoning genes from bovine milk.
Subject(s)
Anti-Bacterial Agents , Coagulase , Microbial Sensitivity Tests , Milk , Staphylococcus , Animals , Milk/microbiology , Cattle , Staphylococcus/genetics , Staphylococcus/drug effects , Staphylococcus/isolation & purification , Staphylococcus/enzymology , Ethiopia , Coagulase/genetics , Coagulase/metabolism , Cross-Sectional Studies , Anti-Bacterial Agents/pharmacology , Staphylococcal Infections/microbiology , Staphylococcal Infections/veterinary , Virulence/genetics , Virulence Factors/genetics , Female , Genes, Bacterial/genetics , Mastitis, Bovine/microbiologyABSTRACT
Bacterial adhesins are cell-surface proteins that anchor to the cell wall of the host. The first stage of infection involves the specific attachment to fibrinogen (Fg), a protein found in human blood. This attachment allows bacteria to colonize tissues causing diseases such as endocarditis. The study of this family of proteins is hence essential to develop new strategies to fight bacterial infections. In the case of the Gram-positive bacterium Staphylococcus aureus, there exists a class of adhesins known as microbial surface components recognizing adhesive matrix molecules (MSCRAMMs). Here, we focus on one of them, the clumping factor A (ClfA), which has been found to bind Fg through the dock-lock-latch mechanism. Interestingly, it has recently been discovered that MSCRAMM proteins employ a catch-bond to withstand forces exceeding 2 nN, making this type of interaction as mechanically strong as a covalent bond. However, it is not known whether this strength is an evolved feature characteristic of the bacterial protein or is typical only of the interaction with its partner. Here, we combine single-molecule force spectroscopy, biophysical binding assays, and molecular simulations to study the intrinsic mechanical strength of ClfA. We find that despite the extremely high forces required to break its interactions with Fg, ClfA is not by itself particularly strong. Integrating the results from both theory and experiments we dissect contributions to the mechanical stability of this protein.
Subject(s)
Coagulase , Fibrinogen , Staphylococcus aureus , Staphylococcus aureus/metabolism , Staphylococcus aureus/chemistry , Coagulase/metabolism , Coagulase/chemistry , Fibrinogen/chemistry , Fibrinogen/metabolism , Protein Binding , Adhesins, Bacterial/metabolism , Adhesins, Bacterial/chemistry , Humans , Protein StabilityABSTRACT
Milk is a source of essential nutrients, but food safety across the milk supply chain has emerged as an integral part of food trade. Of the several food safety hazards, antimicrobial-resistant Staphylococcus species have emerged as one of the major microbial hazards with significant public health concerns. The present crosssectional study was undertaken with the objective to isolate Staphylococcus species from the milk supply chain, characterize isolates for antimicrobial resistance, and trace the origin of isolates using molecular techniques. Samples collected from the formal and informal milk supply chains showed prevalence of Staphylococcus species of 4.3% (n=720); isolates were identified as coagulase-positive (S. aureus 67.7% and S. intermedius 6.4%) and coagulase-negative (S. lentus 9.6%, S. sciuri 3.2%, S. xylosus 3.2%, S. schleiferi 3.2%, S. felis 3.2%, and S. gallinarum 3.2%) species. Staphylococcus isolates showed antimicrobial resistance to methicillin (32.2%), ß-lactam (41.9%), and macrolide-lincosamide-streptogramin B (3.2%). Staphylococcus isolates phenotypically resistant to methicillin also carried the mecA gene and displayed diverse pulsed field gel electrophoresis (PFGE) profiles, indicating their diverse origins in the milk supply chain. Based on the similarity of PFGE profile, the origin of one of the Staphylococcus isolates was traced to the soil in contact with milch cows. The findings of this study highlight the need for more comprehensive microbial risk analysis studies across the milk supply chain, capacity building, creation of awareness among stakeholders about the judicious use of antimicrobials, and protection of public health using a One-Health approach.
Subject(s)
Anti-Bacterial Agents , Milk , Staphylococcus , Milk/microbiology , Animals , Staphylococcus/drug effects , Staphylococcus/genetics , Staphylococcus/isolation & purification , Anti-Bacterial Agents/pharmacology , Microbial Sensitivity Tests , Staphylococcal Infections/microbiology , Staphylococcal Infections/epidemiology , Food Microbiology , Humans , Cattle , Bacterial Proteins/genetics , Coagulase/genetics , Coagulase/metabolism , Drug Resistance, Bacterial/geneticsABSTRACT
Coagulase-negative Staphylococcus (CoNS) species inhibiting Staphylococcus aureus has been described in the skin of atopic dermatitis (AD) patients. This study evaluated whether Staphylococcus spp. from the skin and nares of AD and non-AD children produced antimicrobial substances (AMS). AMS production was screened by an overlay method and tested against NaOH, proteases and 30 indicator strains. Clonality was assessed by pulsed-field gel electrophoresis. Proteinaceous AMS-producers were investigated for autoimmunity by the overlay method and presence of bacteriocin genes by polymerase chain reaction. Two AMS-producers had their genome screened for AMS genes. A methicillin-resistant S. aureus (MRSA) produced proteinaceous AMS that inhibited 51.7% of the staphylococcal indicator strains, and it was active against 60% of the colonies selected from the AD child where it was isolated. On the other hand, 57 (8.8%) CoNS from the nares and skin of AD and non-AD children, most of them S. epidermidis (45.6%), reduced the growth of S. aureus and other CoNS species. Bacteriocin-related genes were detected in the genomes of AMS-producers. AMS production by CoNS inhibited S. aureus and other skin microbiota species from children with AD. Furthermore, an MRSA colonizing a child with AD produced AMS, reinforcing its contribution to dysbiosis and disease severity.
Subject(s)
Coagulase , Dermatitis, Atopic , Methicillin-Resistant Staphylococcus aureus , Microbiota , Skin , Staphylococcus , Dermatitis, Atopic/microbiology , Humans , Methicillin-Resistant Staphylococcus aureus/genetics , Skin/microbiology , Child , Coagulase/genetics , Coagulase/metabolism , Staphylococcus/genetics , Bacteriocins/genetics , Anti-Bacterial Agents/pharmacology , Child, Preschool , Microbial Sensitivity TestsABSTRACT
BACKGROUND: Recently, linezolid-resistant staphylococci have become an emerging problem worldwide. Understanding the mechanisms of resistance, molecular epidemiology and transmission of linezolid-resistant CoNS in hospitals is very important. METHODS: The antimicrobial susceptibilities of all isolates were determined by the microdilution method. The resistance mechanisms and molecular characteristics of the strains were determined using whole-genome sequencing and PCR. RESULTS: All the strains were resistant to oxacillin and carried the mecA gene; 13 patients (36.1%) had prior linezolid exposure. Most S. epidermidis and S. hominis isolates were ST22 and ST1, respectively. MLST typing and evolutionary analysis indicated most linezolid-resistant CoNS strains were genetically related. In this study, we revealed that distinct CoNS strains have different mechanisms of linezolid resistance. Among ST22-type S. epidermidis, acquisition of the T2504A and C2534T mutations in the V domain of the 23 S rRNA gene, as well as mutations in the ribosomal proteins L3 (L101V, G152D, and D159Y) and L4 (N158S), were linked to the development of linezolid resistance. In S. cohnii isolates, cfr, S158Y and D159Y mutations in the ribosomal protein L3 were detected. Additionally, emergence of the G2576T mutation and the cfr gene were major causes of linezolid resistance in S. hominis isolates. The cfr gene, G2576T and C2104T mutations, M156T change in L3 protein, and I188S change in L4 protein were found in S. capitis isolates. CONCLUSION: The emergence of linezolid-resistant CoNS in the environment is concerning because it involves clonal dissemination and frequently coexists with various drug resistance mechanisms.
Subject(s)
Anti-Bacterial Agents , Linezolid , Microbial Sensitivity Tests , Staphylococcal Infections , Tertiary Care Centers , Linezolid/pharmacology , Humans , China/epidemiology , Staphylococcal Infections/microbiology , Staphylococcal Infections/epidemiology , Anti-Bacterial Agents/pharmacology , Female , Male , Middle Aged , Multilocus Sequence Typing , Aged , Whole Genome Sequencing , Staphylococcus/drug effects , Staphylococcus/genetics , Staphylococcus/classification , Staphylococcus/enzymology , Coagulase/metabolism , Coagulase/genetics , RNA, Ribosomal, 23S/genetics , Adult , Methicillin Resistance/genetics , Mutation , Bacterial Proteins/geneticsABSTRACT
Pet food have been considered as possible vehicles of bacterial pathogens. The sudden boom of the pet food industry due to the worldwide increase in companion animal ownership calls for pet food investigations. Herein, this study aimed to determine the frequency, antimicrobial susceptibility profile, and molecular characteristics of coagulase-negative staphylococci (CoNS) in different pet food brands in Brazil. Eighty-six pet food packages were screened for CoNS. All isolates were identified at species level by MALDI-TOF MS and species-specific PCR. Antimicrobial susceptibility testing was performed by disc diffusion and broth microdilution (vancomycin and teicoplanin only) methods. The D-test was used to screen for inducible clindamycin phenotype (MLS-B). SCCmec typing and detection of mecA, vanA, vanB, and virulence-encoding genes were done by PCR. A total of 16 (18.6 %) CoNS isolates were recovered from pet food samples. Isolates were generally multidrug-resistant (MDR). All isolates were completely resistant (100 %) to penicillin. Resistances (12.5 % - 75 %) were also observed for fluoroquinolones, sulfamethoxazole-trimethoprim, tetracycline, rifampicin, erythromycin, and tobramycin. Isolates were susceptible to vancomycin (MICs <0.25-1 µg/mL) and teicoplanin (MICs <0.25-4 µg/mL). Intriguingly, 3/8 (37.5 %) CoNS isolates with the ERYRCLIS antibiotype expressed MLS-B phenotype. All isolates harboured blaZ gene. Seven (43.8 %) isolates carried mecA; and among them, the SCCmec Type III was the most frequent (n = 5/7; 71.4 %). Isolates also harboured seb, see, seg, sej, sem, etb, tsst, pvl, and hla toxin virulence-encoding genes (6.3 % - 25 %). A total of 12/16 (75 %) isolates were biofilm producers, while the icaAB gene was detected in an S. pasteuri isolate. Herein, it is shown that pet food is a potential source of clinically important Gram-positive bacterial pathogens. To the best of our knowledge, this is the first report of MLS-B phenotype and MR-CoNS in pet food in Latin America.
Subject(s)
Anti-Bacterial Agents , Clindamycin , Coagulase , Microbial Sensitivity Tests , Staphylococcus , Staphylococcus/drug effects , Staphylococcus/genetics , Staphylococcus/isolation & purification , Brazil , Anti-Bacterial Agents/pharmacology , Coagulase/metabolism , Animals , Clindamycin/pharmacology , Methicillin/pharmacology , Animal Feed/microbiology , Food Microbiology , Pets/microbiology , Drug Resistance, Multiple, Bacterial/geneticsABSTRACT
This study was condcuted to examine the association of area under the curve (AUC)/minimum inhibitory concentration (MIC) and trough concentration (Ctrough) of vancomycin with treatment outcome and nephrotoxicity in infections caused by Enterococcus spp. and coagulase-negative Staphylococci (CoNS). Peak and trough concentrations were used to calculate AUC in 89 patients receiving vancomycin for infections with Enterococcus spp. (n = 65) or CoNS (n = 24). Correlations between Ctrough, AUC/MIC, early clinical response (ECR), and nephrotoxicity were assessed and cutoff values were determined. Sixty-three (70.8%) patients showed improvement in ECR and 10 (11.2%) experienced nephrotoxicity. Enterococcus spp. infections displayed correlations between AUC/MIC and ECR for AUC0-24 h/MIC (r2 = 0.27, P ≤ .05) and AUC24-48 h/MIC (r2 = 0.28, P ≤ .05), but not for Ctrough (r2 = 0.21, P > .05). There were no correlations between Ctrough (r2 = 0.26, P > .05), AUC0-24 h/MIC (r2 = -0.12, P > .05), AUC24-48 h/MIC (r2 = 0.01, P > .05) and ECR for CoNS. In the CoNS group, a moderate correlation was found between ECR and Ctrough at a cutoff value of 6.9 µg/mL. In addition, nephrotoxicity is also moderately associated with AUC0-24 h and AUC24-48 h at 505.7 and 667.1 µgâ¢h/mL, respectively. A strong correlation between nephrotoxicity and Ctrough was observed when the cutoff value was 18.9 µg/mL. AUC/MIC during the first 48 h was a determinant of vancomycin efficacy in Enterococcus infections but not for CoNS. Ctrough was not correlated with clinical outcome. Nephrotoxicity could be predicted using Ctrough and AUC for infections with both pathogens.
Subject(s)
Anti-Bacterial Agents , Area Under Curve , Enterococcus , Microbial Sensitivity Tests , Staphylococcal Infections , Staphylococcus , Vancomycin , Humans , Male , Female , Vancomycin/pharmacokinetics , Vancomycin/adverse effects , Vancomycin/blood , Vancomycin/administration & dosage , Vancomycin/therapeutic use , Anti-Bacterial Agents/pharmacokinetics , Anti-Bacterial Agents/adverse effects , Anti-Bacterial Agents/administration & dosage , Anti-Bacterial Agents/blood , Anti-Bacterial Agents/therapeutic use , Middle Aged , Staphylococcus/drug effects , Prospective Studies , Aged , Staphylococcal Infections/drug therapy , Staphylococcal Infections/microbiology , Enterococcus/drug effects , Adult , Gram-Positive Bacterial Infections/drug therapy , Coagulase/metabolism , Treatment Outcome , Aged, 80 and overABSTRACT
PURPOSE: Coagulase-negative staphylococci (CoNS) are Gram-positive organisms that are a known component of normal skin flora and the most common cause of nosocomial bacteremia. For CoNS species, the vancomycin MIC breakpoint for susceptibility set by the Clinical and Laboratory Standards Institute is ≤4 µg/mL. There has been published reports of vancomycin heteroresistance in CoNS with vancomycin MICs of 2 to 4 µg/mL. The aim of this retrospective cohort analysis was to assess the clinical impact of vancomycin MICs <2 µg/mL versus ≥2 µg/mL in adult patients with CoNS bloodstream infections. METHODS: Adult patients admitted to University Medical Center New Orleans with a blood culture positive for CoNS were assessed. The primary outcome was difference in 30-day mortality. Secondary outcomes were in-hospital, all-cause mortality; duration of bacteremia; hospital length of stay; and percentage of oxacillin-resistant CoNS. FINDINGS: There was no difference in mortality in the vancomycin MIC <2 µg/mL group versus the vancomycin MIC ≥2 µg/mL group at 30 days (15.4% vs 17.4%; P = 1). In-hospital, all-cause mortality was also not different between groups (11.5% vs 13%; P = 1). Hospital length of stay between groups was 28.2 days versus 21 days (P = 0.692). Median duration of bacteremia was 1 day in both groups (P = 0.975), and median scheduled duration of antibiotic therapy was 14.9 days and 19.5 days (P = 0.385). The source and mode of acquisition of CoNS were similar between groups. Of all CoNS isolates, 58.7% (44 of 75) were oxacillin resistant. Staphylococcus epidermidis was the most common CoNS species at 66.7% (50 of 75). Of all isolates, 30.7% (23 of 75) had a vancomycin MIC ≥2 µg/mL, and 87% (20 of 23) of these were S. epidermidis. There was a higher percentage of S. epidermidis in the vancomycin MIC ≥2 µg/mL group than in the MIC <2 µg/mL group (87% vs 57.7%; P = 0.012). CoNS with a vancomycin MIC ≥2 µg/mL were also more likely to be oxacillin resistant (78.3% vs 50%; P = 0.005). IMPLICATIONS: There was no difference in clinical outcomes in adult patients with a CoNS bloodstream infection with a vancomycin MIC <2 µg/mL versus ≥2 µg/mL. At present, vancomycin remains appropriate empiric therapy for CoNS bloodstream infection. Further research is needed to determine if there is a true clinical impact of a vancomycin MIC ≥2 µg/mL in CoNS infections.
Subject(s)
Anti-Bacterial Agents , Bacteremia , Coagulase , Microbial Sensitivity Tests , Staphylococcal Infections , Staphylococcus , Vancomycin , Humans , Vancomycin/therapeutic use , Vancomycin/pharmacology , Bacteremia/drug therapy , Bacteremia/microbiology , Bacteremia/mortality , Male , Retrospective Studies , Female , Staphylococcal Infections/drug therapy , Staphylococcal Infections/microbiology , Middle Aged , Anti-Bacterial Agents/therapeutic use , Anti-Bacterial Agents/pharmacology , Coagulase/metabolism , Staphylococcus/drug effects , Staphylococcus/isolation & purification , Aged , Treatment Outcome , Adult , Length of Stay/statistics & numerical dataABSTRACT
The species- and clone-specific susceptibility of Staphylococcus cells for bacteriophages is governed by the structures and glycosylation patterns of wall teichoic acid (WTA) glycopolymers. The glycosylation-dependent phage-WTA interactions in the opportunistic pathogen Staphylococcus epidermidis and in other coagulase-negative staphylococci (CoNS) have remained unknown. We report a new S. epidermidis WTA glycosyltransferase TagE whose deletion confers resistance to siphoviruses such as ΦE72 but enables binding of otherwise unbound podoviruses. S. epidermidis glycerolphosphate WTA was found to be modified with glucose in a tagE-dependent manner. TagE is encoded together with the enzymes PgcA and GtaB providing uridine diphosphate-activated glucose. ΦE72 transduced several other CoNS species encoding TagE homologs, suggesting that WTA glycosylation via TagE is a frequent trait among CoNS that permits interspecies horizontal gene transfer. Our study unravels a crucial mechanism of phage-Staphylococcus interaction and horizontal gene transfer, and it will help in the design of anti-staphylococcal phage therapies.IMPORTANCEPhages are highly specific for certain bacterial hosts, and some can transduce DNA even across species boundaries. How phages recognize cognate host cells remains incompletely understood. Phages infecting members of the genus Staphylococcus bind to wall teichoic acid (WTA) glycopolymers with highly variable structures and glycosylation patterns. How WTA is glycosylated in the opportunistic pathogen Staphylococcus epidermidis and in other coagulase-negative staphylococci (CoNS) species has remained unknown. We describe that S. epidermidis glycosylates its WTA backbone with glucose, and we identify a cluster of three genes responsible for glucose activation and transfer to WTA. Their inactivation strongly alters phage susceptibility patterns, yielding resistance to siphoviruses but susceptibility to podoviruses. Many different CoNS species with related glycosylation genes can exchange DNA via siphovirus ΦE72, suggesting that glucose-modified WTA is crucial for interspecies horizontal gene transfer. Our finding will help to develop antibacterial phage therapies and unravel routes of genetic exchange.
Subject(s)
Staphylococcal Infections , Staphylococcus epidermidis , Humans , Staphylococcus epidermidis/genetics , Staphylococcus epidermidis/metabolism , Staphylococcus aureus/genetics , Coagulase/metabolism , Glucose/metabolism , Teichoic Acids/metabolism , Staphylococcus/metabolism , Staphylococcus Phages/genetics , DNA/metabolism , Cell Wall/metabolism , Staphylococcal Infections/metabolismABSTRACT
Since September 2018, serious meningitis has been found on some breeding-duck farms in Shandong Province, China. A large number of ducks exhibit severe neurological symptoms. The ducks were randomly selected for laboratory testing. Duck brain samples were collected using standard sterile techniques, and the staphylococci isolates were detected in 404 (70.14%) out of 576 brain samples. A total of 525 coagulase-negative staphylococci (CoNS) strains were isolated, including 6 species: Staphylococcus sciuri (S. sciuri) (67.24%, 353/525), Staphylococcus epidermidis (S. epidermidis) (9.71%, 51/525), Staphylococcus saprophyticus (S. saprophyticus) (8.38%, 44/525), Staphylococcus lentus (S. lentus) (7.62%, 40/525), Staphylococcus haemolyticus (S. haemolyticus) (2.48%, 13/525), and Staphylococcus xylosus (S. xylosus) (4.57%, 24/525). Mixed strain infections were detected in 121 (29.95%) infected presentations. The antimicrobial susceptibility testing indicated that 40.38% of the isolates exhibited multi-drug resistance, and 53.90% of the strains were methicillin-resistant strains by amplification of the methicillin resistance gene (mecA) gene. Through experimental reproduction of the disease, we determined that the CoNS strains were the leading pathogens causing bacterial meningitis in ducks. Although these CoNS strains does not directly cause the death of sick ducks, they still cause large economic losses due to the retarded growth and development of the sick ducks, lower feed returns, and lower grades of processed duck products. The results of this study will contribute to our understanding of the epidemiology and pathogenesis of CoNS and be helpful in the prevention and treatment of the infection.
Subject(s)
Coagulase , Ducks , Meningitis, Bacterial , Poultry Diseases , Staphylococcal Infections , Staphylococcus , Animals , Staphylococcus/drug effects , Staphylococcus/isolation & purification , Staphylococcus/enzymology , Poultry Diseases/microbiology , Poultry Diseases/epidemiology , Staphylococcal Infections/veterinary , Staphylococcal Infections/microbiology , Staphylococcal Infections/epidemiology , Coagulase/metabolism , Meningitis, Bacterial/veterinary , Meningitis, Bacterial/microbiology , Meningitis, Bacterial/epidemiology , China/epidemiology , Anti-Bacterial Agents/pharmacologyABSTRACT
Staphylococcus schleiferi and Staphylococcus coagulans are opportunistic pathogens of animals and humans. They were previously classified as Staphylococcus schleiferi subs. schleiferi and Staphylococcus schleiferi subs. coagulans, respectively, and recently reclassified as separate species. S. coagulans, is frequently associated with dogs, whereas S. schleiferi is more commonly isolated from humans. Coagulase activity status is a defining characteristic of the otherwise closely related species. However, the use of coagulase tests originally developed to distinguish S. aureus from non-coagulase-producing staphylococci, for this purpose is questionable and the basis for their host preference has not been elucidated. In the current study, a putative coa gene was identified and correlated with coagulase activity measured using a chromogenic assay with human and bovine prothrombin (closely related to canine prothrombin). The results of the tests performed with human prothrombin showed greater reactivity of S. coagulans isolates from humans than isolates obtained from dogs with the same substrate. Our data suggest that unlike S. coagulans isolates from humans, isolates from dogs have more coagulase activity with bovine prothrombin (similar to canine prothrombin) than human prothrombin. Differences in nuc and 16s rRNA genes suggest a divergence in S. coagulans and S. schleiferi. Phenotypic and genotypic variation based on the number of IgG binding domains, and the numbers of tandem repeats in C-terminal fibronectin binding motifs was also found in protein A, and fibronectin-binding protein B respectively. This study identified a coa gene and associated phenotypic activity that differentiates S. coagulans and S. schleiferi and identified key phylogenetic and phenotypic differences between the species.
Subject(s)
Dog Diseases , Staphylococcal Infections , Animals , Humans , Dogs , Cattle , Staphylococcus aureus/genetics , Staphylococcus aureus/metabolism , Coagulase/genetics , Coagulase/metabolism , RNA, Ribosomal, 16S/genetics , Fibronectins/genetics , Phylogeny , Prothrombin , Staphylococcus/metabolism , Staphylococcal Infections/veterinaryABSTRACT
BACKGROUND: Venous access device-related bloodstream infection (VAD-BSI) with coagulase-negative staphylococci (CoNS) is a common complication after allogeneic hematopoietic cell transplantation (alloHCT). Standard systemic antimicrobial therapy for uncomplicated VAD-BSI with methicillin-resistant CoNS consists of intravenous (IV) vancomycin (vanco). This requires hospitalization, needs new competent venous access, exposes patients to potential toxicity (mainly renal) and increases the risk of commensal flora dysbiosis with selection of vanco-resistant enterococci. Combined with VAD management (removal or antibiotic locks), oral minocycline (mino) has been evaluated as an alternative systemic therapy for the treatment of uncomplicated VAD-BSIs with CoNS at our center, primarily when the reference treatment with IV vanco was not possible (renal failure or allergy) or when hospitalization was refused by patients. Here, we retrospectively report our single center experience with this mino-based approach. PATIENTS AND METHODS: From January 2012 to December 2020, 24 uncomplicated VAD-BSIs with CoNS in 23 alloHCT patients were treated with oral mino as systemic antibiotic therapy in combination with VAD management. VAD were implantable ports (n = 17), tunneled catheter (n = 1) or PIC-lines (n = 6). Staphylococci were S. epidermidis (n = 21) or S. haemolyticus (n = 3). Mino was administered with a loading dose of 200 mg followed by 100 mg BID for 7-14 days. For 8 VAD-BSIs, patients were initially treated with IV vanco for the first 1-3 days followed by oral mino, while 16 VAD-BSIs were treated with oral mino as the sole antimicrobial agent for systemic therapy. VAD management consisted of catheter removal (for tunneled catheters and PIC-lines, n = 7) or antibiotic locks with vanco (n = 15) or gentamicin (n = 2) administered at least 3 times a week for 14 days (for ports). RESULTS: Overall, clearance of bacteremia (as assessed by negativity for the same CoNS of surveillance peripheral blood cultures drawn between day+ 3 and +30 after initiation of systemic therapy) was achieved in all but 1 patient (with port) who had persistent bacteremia at day +9. No complication such as suppurative thrombophlebitis, endocarditis, distant foci of infection or BSI-related death was observed in any patient during the 3-month period after initiation of treatment. Regarding the 17 port-BSI cases for which VAD conservative strategy was attempted, failure of 3-month VAD preservation was documented in 7/17 cases and 3-month recurrence of VAD-BSI was observed in 3/17 cases (with 1 patient with cellulitis). Treatment with mino was well tolerated except for a mild skin rash in one patient. CONCLUSION: Further prospective studies are needed to evaluate efficacy and safety of this approach.
Subject(s)
Bacteremia , Catheter-Related Infections , Hematopoietic Stem Cell Transplantation , Staphylococcal Infections , Humans , Minocycline/therapeutic use , Coagulase/metabolism , Coagulase/therapeutic use , Staphylococcal Infections/drug therapy , Staphylococcal Infections/etiology , Retrospective Studies , Catheter-Related Infections/drug therapy , Catheter-Related Infections/epidemiology , Staphylococcus/metabolism , Anti-Bacterial Agents/adverse effects , Vancomycin/therapeutic use , Bacteremia/drug therapy , Bacteremia/etiology , Bacteremia/epidemiology , Hematopoietic Stem Cell Transplantation/adverse effectsABSTRACT
Staphylococcus aureus (S. aureus) induces a variety of infectious diseases in humans and animals and is responsible for hospital- and community-acquired infections. The aim of this study was to investigate how bilobetin, a natural compound, attenuates S. aureus virulence by inhibiting two key virulence factors, von Willebrand factor-binding protein (vWbp) and staphylocoagulase (Coa). The results showed that bilobetin inhibited Coa- or vWbp-induced coagulation without affecting S. aureus proliferation. The Western blotting and fluorescence quenching assays indicated that bilobetin did not affect the expression of vWbp and Coa but directly bound to the proteins with KA values of 1.66 × 104 L/mol and 1.04 × 104 L/mol, respectively. To gain further insight into the mechanism of interaction of bilobetin with these virulence factors, we performed molecular docking and point mutation assays, which indicated that the TYR-6 and TYR-18 residues on vWbp and the ALA-190 and ASP-189 residues on Coa were essential for the binding of bilobetin. In addition, the in vivo studies showed that bilobetin ameliorated lung tissue damage and inflammation caused by S. aureus, thereby improving the survival of mice. Furthermore, the use of bilobetin as an adjuvant in combination with vancomycin was more effective in the treatment of a mouse model of pneumonia. Taken together, bilobetin had a dual inhibitory effect on vWbp and Coa by reducing the virulence of S. aureus, suggesting that it is a viable lead compound against S. aureus infections.
Subject(s)
Coagulase , Staphylococcal Infections , Humans , Mice , Animals , Coagulase/genetics , Coagulase/metabolism , Coagulase/pharmacology , Carrier Proteins/metabolism , Staphylococcus aureus , Virulence , von Willebrand Factor/metabolism , von Willebrand Factor/pharmacology , Molecular Docking Simulation , Staphylococcal Infections/drug therapy , Virulence Factors/genetics , Virulence Factors/metabolismABSTRACT
The massive use of antibiotics has resulted in an alarming increase in antibiotic resistance in Staphylococcus aureus (S. aureus). This study aimed to identify the inhibitory effect of salicin on S. aureus. Coagulase (Coa) activity was assessed using inâ vitro Coa assays and Western blot, thermal shift assay (TSA), fluorescence quenching and molecular docking experiments were conducted to verify the interaction between salicin and Coa. An inâ vivo mouse pneumonia model demonstrated that salicin can reduce the virulence of S. aureus. In vitro Coa assays elucidated that salicin directly inhibited Coa activity. The Western blot and TSA results suggested that salicin did not block the expression of Coa but affected the thermal stability of the protein by binding to Coa. The fluorescence quenching, molecular docking and molecular dynamics assays have found that the most promising binding site between salicin and Coa was GLN-97. The pneumonia model of mice infected with S. aureus revealed that salicin could not only reduce the content of lung bacteria in mice but also prolong their survival. Salicin was identified as a novel anti-infective candidate compound with the potential to target Coa and inhibit its activity by binding to it, which would facilitate the development of roadmaps for future research.
Subject(s)
Methicillin-Resistant Staphylococcus aureus , Pneumonia , Staphylococcal Infections , Animals , Mice , Staphylococcus aureus , Coagulase/metabolism , Coagulase/pharmacology , Bacterial Proteins , Molecular Docking Simulation , Microbial Sensitivity Tests , Staphylococcal Infections/drug therapy , Staphylococcal Infections/microbiology , Anti-Bacterial Agents/pharmacologyABSTRACT
Background: The objective of this study was to isolate Staphylococcus spp. and to characterize the resistance profile in nasal samples from pigs slaughtered for consumption. Material and Methods: Intranasal swabs were collected from 100 pigs immediately after bleeding in a slaughterhouse located in the largest pork production region in Brazil, these samples were cultured and isolated to identify Staphylococcus spp. in coagulase positive (CoPS) and coagulase negative (CoNS) and molecular identification of Staphylococcus aureus and then subjected to the disk-diffusion test to identify the bacterial resistance profile and search for the mecA gene. Results: Of the 100 samples collected, it was possible to isolate 79 Staphylococcus spp., of these, 72.15% were classified as CoNS and 27.85% of the isolates classified as CoPS. Among the CoPS isolates, 77.27% were identified as S. aureus. Through the disk-diffusion test, it was possible to verify isolates resistant to clindamycin and erythromycin (98.73%), chloramphenicol (93.67%), and doxycycline (89.87%). There was amplification of the mecA gene in 30.38% of Staphylococcus spp. Conclusion: The results of this study highlight the need for the careful use of antibiotics in swine production, in addition to aiming at continuous surveillance in relation to the rate of multiresistant microorganisms within these environments, focused on large industrial centers; such results also indicate the importance of understanding, through future studies, possible pathways to transmission of these microorganisms directly, or indirectly, through meat products derived from these pigs, which can be considered neglected diffusers of variants of Staphylococcus spp. resistant to antibiotics or carriers of important resistance genes related to One Health.
Subject(s)
Staphylococcal Infections , Swine Diseases , Animals , Swine , Staphylococcus/genetics , Staphylococcus aureus/genetics , Coagulase/metabolism , Bacterial Proteins/genetics , Microbial Sensitivity Tests/veterinary , Anti-Bacterial Agents/pharmacology , Staphylococcal Infections/epidemiology , Staphylococcal Infections/veterinary , Staphylococcal Infections/microbiology , Swine Diseases/epidemiologyABSTRACT
Aim: Our primary objective was to investigate the protective effects and mechanisms of isovanillic acid in mice infected with Staphylococcus aureus Newman. Methods: In vitro coagulation assays were used to validate vWbp and Coa as inhibitory targets of isovanillic acid. The binding mechanism of isovanillic acid to vWbp and Coa was investigated using molecular docking and point mutagenesis. Importantly, a lethal pneumonia mouse model was used to assess the effect of isovanillic acid on survival and pathological injury in mice. Results & Conclusion: Isovanillic acid reduced the virulence of S. aureus by directly binding to inhibit the clotting activity of vWbp and Coa, thereby reducing lung histopathological damage and improving the survival rate in mice with pneumonia.
Subject(s)
Coagulase , Staphylococcal Infections , Mice , Animals , Coagulase/metabolism , Staphylococcus aureus/metabolism , Molecular Docking Simulation , Staphylococcal Infections/prevention & controlABSTRACT
AIMS: To understand the Staphylococcus coagulans prevalence in causing skin infections in dogs and detection of various virulence genes in Staph. coagulans isolates. METHODS AND RESULTS: Staph. coagulans was isolated from pus swabs collected from dogs with skin infection and identified by detecting thermonuclease, coagulase, and urease genes. The presence of methicillin-resistant gene (mecA) was performed by PCR. Antimicrobial susceptibility test was carried out by disc diffusion method. In total, 38 Staph. coagulans clinical isolates and 42 Staph. coagulans genomes available in NCBI database were screened for 19 virulence genes by PCR and in silico prediction, respectively. A prevalence of 13.8% (38/275) of Staph. coagulans dog skin infection was observed and 15.8% (6/38) of Staph. coagulans isolates carried mecA gene. Many Staph. coagulans isolates were susceptible to all tested antimicrobials. Twenty nine per cent isolates were resistant to ciprofloxacin. Genes encoding leukotoxins, DNase, exfoliative toxin, superantigen-like exotoxin, immunoglobulin-binding proteins, fibrinogen-binding proteins, autolysin, and rod shape-determining protein were detected in almost all the Staph. coagulans clinical isolates and genomes from NCBI database, whereas anti-adhesin plasma-sensitive protein genes were present in relatively lesser number of Staph. coagulans clinical isolates and genomes from NCBI database. CONCLUSIONS: Staph. coagulans possesses many virulence factors that are present in other coagulase-positive staphylococci, such as Staph. aureus and Staph. pseudintermedius. The presence of two bi-component leukotoxin genes in tandem with other virulence factor genes in a single pathogenic island in the Staph. coagulans genomes explained their eminence in the virulence of Staph. coagulans causing infections. Staph. coagulans was classified as a separate species in the year 2020 and primarily causes skin infections in dogs. Identification of this species is not included in any of the automated bacterial identification systems. Hence, many veterinary laboratories do not have a strategy to identify this bacterium. This study will help in the identification of Staph. coagulans in veterinary laboratories by PCR apart from detecting various virulence factors present in this pathogen. The existence of many virulence factors and prevalence in different animals in varied geographical locations suggest that Staph. coagulans is an important coagulase-positive staphylococcal pathogen in animals.
Subject(s)
Staphylococcal Infections , Virulence Factors , Animals , Dogs , Virulence Factors/genetics , Coagulase/genetics , Coagulase/metabolism , Staphylococcus , Staphylococcus aureus , Staphylococcal Infections/veterinary , Staphylococcal Infections/microbiology , Anti-Bacterial Agents/pharmacology , Microbial Sensitivity TestsABSTRACT
Technological, safety-related and volatile properties were analyzed in coagulase-negative staphylococci (CNS) isolates from Chinese spontaneously fermented meat products. A total of 107 CNS isolates were identified via 16 S rRNA sequencing, and the most recovered species were S. saprophyticus (53.3 %), S. edaphicus (12.1 %), and S. epidermidis (10.3 %). Among them, 58 CNS isolates belonging to 9 species were selected with higher activities of catalase, nitrate reductase, proteolysis, and lipolysis, as well as higher tolerance to stressful environmental conditions. Then, 7 CNS isolates belonging to 4 species were further selected based upon excellent technological characteristics, lack of hemolysis and antibiotic resistance, and a low production of biogenic amines. The volatile profiles of these 7 strains cultivated in pork broth was determined. S. casei No. 1 produced significant amounts of phenethyl alcohol, geraniol, and 3-methyl-butanol. S. xylosus No. 120 produced the highest amount of methyl ketones with the potential to provide dry-cured odor of fermented meats. The volatile profile was highly strain dependent. Several CNS identified in this study have the potential to be used as the starter cultures for fermented meat products.
Subject(s)
Meat Products , China , Coagulase/metabolism , Fermentation , Food Microbiology , Staphylococcus/genetics , Staphylococcus/metabolismABSTRACT
Coagulase-negative staphylococci (CNS) are one of the most pervasive heterogeneous groups of bacteria which are used as starter/adjunct cultures to enhance the aroma and texture of fermented foods. The organoleptic characteristics of fermented foods rely on disparate metabolic attributes of CNS. Nitrate reductase production from CNS improves sensory characteristics of foods by converting nitrate into nitrite. These bacteria utilize arginine via arginine deiminase pathway in the cytosol, and thus, play effective role in the generation of colour of fermented foods. Coagulase-negative Staphylococcus spp. develop flavour in foods by fermenting carbohydrates, converting amino acids, inducing ß-oxidation of lipids, and secreting esterases. Additionally, the characteristic flavour of foods depends on the proteolytic and lipolytic properties of CNS strains too. Coagulase-negative staphylococci strains have revealed exemplary functional or probiotic traits by showing tolerance to acidic pH and bile, depicting adhesion characteristics, producing exopolysaccharide, and secreting therapeutic bacteriocins. Unfortunately, some CNS strains have shown antibiotics resistance, enterotoxins secretions, biogenic amine productions, haemolytic activities, and biofilm formations, thereby indicated the utilization of CNS on strain-by-strain basis. This review sheds light not only on the metabolic heterogeneity and techno-functional traits but also the safety and pathogenic aspects of fermented foods-associated CNS strains.