RESUMO
The antibiotic resistance among coagulase - negative Staphylococcus (CoNS) species towards methicillin is rarely reported in veterinary medicine. Under the aspect/concept of One Health, those strains pose a risk to human health due to the presence in canine pets where the transfer of resistant genetic markers might occur to other staphylococci species. The aim of this study was to investigate the antimicrobial resistance pattern among Coagulase Negative Staphylococci (CoNS) isolated from asymptomatic dogs and those affected by topic infections. Swabs from 254 dogs were first seeded in Mannitol Salt Agar. Species identification was conducted by Matrix Assisted Laser Desorption ionization - time of flight (MALDI-TOF ms) as previously described. The susceptibility test was performed by disk diffusion according to CLSI standards. Detection of mecA gene was performed. CoNS could be recovered from both groups of dogs and an alarming presence of methicillin-resistant coagulase negative staphylococci (MRCoNS) was confirmed, in 10.2% (17/166) of the samples. Eight of those methicillin resistant strains were isolated from asymptomatic dogs whereas nine were present in dogs affected by pyoderma and otitis externa.
Assuntos
Coagulase/deficiência , Reservatórios de Doenças/microbiologia , Doenças do Cão/microbiologia , Resistência a Meticilina , Infecções Estafilocócicas/veterinária , Staphylococcus/isolamento & purificação , Animais , Antibacterianos/farmacologia , Doenças do Cão/transmissão , Cães , Farmacorresistência Bacteriana , Genes Bacterianos , Humanos , Resistência a Meticilina/genética , Testes de Sensibilidade Microbiana , Proteínas de Ligação às Penicilinas , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/transmissão , Staphylococcus/classificação , Staphylococcus/enzimologia , Staphylococcus/genéticaRESUMO
Staphylococcus chromogenes is one of the main coagulase-negative staphylococci isolated from mastitis of dairy cows. We describe S. chromogenes isolates that can clot plasma. Since the main pathogen causing mastitis is coagulase-positive Staphylococcus aureus, the coagulase-positive phenotype of S. chromogenes described here can easily lead to misidentification.
Assuntos
Coagulação Sanguínea , Coagulase/deficiência , Plasma/metabolismo , Staphylococcus/metabolismo , Animais , Bovinos , Mastite Bovina/diagnóstico , Mastite Bovina/microbiologia , Staphylococcus/classificação , Staphylococcus/isolamento & purificaçãoRESUMO
INTRODUCTION: Bovine mastitis is a frequent cause of economic loss in dairy herds. Coagulase-negative staphylococci (CoNS) are increasing in importance as causeof bovine intramammary infection (IMI) throughout the world in recent years. CoNShave been isolated from milk samples collected from cows with clinical andsubclinical mastitis in several countries. Identification of mastitis pathogensis important when selection appropriate antimicrobial therapy. METHODOLOGY: A total of 93 strains of Staphylococcusspp isolated from bovine clinical andsubclinical mastitis in Argentina during 2010-2013 were identified by Restriction Fragment Length Polymorphism (RFLP)-PCR using the gap gene. The isolates were tested by PCR for the presence of blaZ gene and mecA gene and were tested by disk diffusion for the susceptibilityto penicillin and cefoxitin. RESULTS: The most common CoNS species was S.chromogenes 46.2% (43/93), followed by S. devriesei 11.8% (11/93) and S. haemolyticus 9.7% (9/93). The blaZ gene was detected in 19 (20.4%) but only 16 (17.2%) isolates were resistant to penicillin; the mecA was detected in6 (6.5%) isolates but only 4 (4.3) were resistant to cefoxitin. The 6 mecA-positive isolates showed oxaxillinMICs ≤ 0.5 µg/ml. DISCUSSION: CoNSare important minor mastitis pathogens and can be the cause of substantial economic losses. Despite the low resistance to PEN in Argentina, the presenceof MR isolates found in this study emphasize the importance of identificationof CoNS when an IMI is present because of the potentially risk of lateraltransfer of resistance genes between staphylococcal species.
Assuntos
Coagulase/deficiência , Mastite Bovina/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus/isolamento & purificação , Resistência beta-Lactâmica , Animais , Argentina , Bovinos , Feminino , Genes Bacterianos , Testes de Sensibilidade Microbiana , Tipagem Molecular , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Infecções Estafilocócicas/microbiologia , Staphylococcus/efeitos dos fármacos , Staphylococcus/enzimologia , Staphylococcus/genéticaRESUMO
INTRODUCTION: Healthcare waste (HCW) might potentially harbor infective viable microorganisms in sanitary landfills. We investigated the antimicrobial susceptibility patterns and the occurrence of the mecA gene in coagulase-negative Staphylococcus strains (CoNS) recovered from the leachate of the HCW in an untreated sanitary landfill. METHODOLOGY: Bacterial identification was performed by physiological and molecular approaches, and minimum inhibitory concentrations (MICs) of antimicrobial drugs were determined by the agar dilution method according to CLSI guidelines. All oxacillin-resistant bacteria were screened for the mecA gene. RESULTS: Out of 73 CoNS, seven different species were identified by 16S rDNA sequencing: Staphylococcus felis (64.4%; n = 47), Staphylococcus sciuri (26.0%; n = 19), Staphylococcus epidermidis (2.7%; n = 2), Staphylococcus warneri (2.7%; n = 2), Staphylococcus lentus (1.4%; n = 1), Staphylococcus saprophyticus (1.4%; n = 1), and Staphylococcus haemolyticus (1.4%; n = 1). Penicillin was the least effective antimicrobial (60.3% of resistance; n = 44) followed by erythromycin (39.8%; n = 29), azithromycin (28.8%; n = 21), and oxacillin (16.5%; n = 12). The most effective drug was vancomycin, for which no resistance was observed, followed by gentamicin and levofloxacin, for which only intermediate resistance was observed (22%, n = 16 and 1.4%, n = 1, respectively). Among the oxacillin-resistant strains, the mecA gene was detected in two isolates. CONCLUSIONS: Considering the high antimicrobial resistance observed, our results raise concerns about the survival of putative bacterial pathogens carrying important resistance markers in HCW and their environmental spread through untreated residues discharged in sanitary landfills.
Assuntos
Coagulase/deficiência , Farmacorresistência Bacteriana Múltipla , Microbiologia Ambiental , Eliminação de Resíduos de Serviços de Saúde/métodos , Infecções Estafilocócicas/transmissão , Staphylococcus/isolamento & purificação , Instalações de Eliminação de Resíduos , Antibacterianos/farmacologia , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Humanos , Resíduos de Serviços de Saúde , Testes de Sensibilidade Microbiana , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Infecções Estafilocócicas/microbiologia , Staphylococcus/classificação , Staphylococcus/enzimologia , Staphylococcus/genéticaRESUMO
To characterize 46 methicillin-resistant coagulase-negative staphylococci from Brazilian neonates, we investigated their SCCmec type, susceptibility and clonality. Staphylococcus epidermidis and Staphylococcus haemolyticus were the prevalent species. SCCmec types III or IV were detected in 53.3% S. epidermidis, whereas 63.6% S. haemolyticus were nontypeable. Despite the diversity, specific clones carried specific SCCmec elements, highlighting that effective typing can help in epidemiological analysis.
Assuntos
Coagulase/deficiência , Genes Bacterianos , Resistência a Meticilina , Infecções Estafilocócicas/microbiologia , Staphylococcus/efeitos dos fármacos , Staphylococcus/genética , Brasil , Variação Genética , Genótipo , Humanos , Recém-Nascido , Staphylococcus/classificação , Staphylococcus/isolamento & purificaçãoRESUMO
The occurrence, resistance phenotype and molecular mechanisms of resistance of methicillin-resistant staphylococci from groin swabs of 109 clinically healthy dogs in Nsukka, Nigeria were investigated. The groin swab samples were cultured on mannitol salt agar supplemented with 10 µg of cloxacillin. Sixteen methicillin-resistant coagulase negative staphylococci (MRCoNS), all harbouring the mecA gene were isolated from 14 (12.8%) of the 109 dogs studied. The MRCoNS isolated were: S. sciuri subspecies rodentium, S. lentus, S. haemolyticus, and S. simulans with S. sciuri subspecies rodentium (62.5%) being the predominant species. Thirteen (81.3%) of the MRCoNS were resistant to tetracycline while 12 (75%) and 10 (62.5%) were resistant to kanamycin and trimthoprim-sulphamethoxazole respectively. None of the isolates was resistant to fusidic acid, linezolid and vancomycin. Thirteen (81.3%) of the MRCoNS were multi-drug resistance (MDR). Other antimicrobial genes detected were: blaZ, tet(K), tet(M), tet(L), erm(B), lnu(A), aacA-aphD, aphA3, str, dfr(G), cat pC221 , and cat pC223 . Methicillin-resistant staphylococci are common colonizers of healthy dogs in Nigeria with a major species detected being S. sciuri subsp. rodentium.
Assuntos
Portador Sadio/veterinária , Coagulase/deficiência , Resistência a Meticilina , Infecções Estafilocócicas/veterinária , Staphylococcus/classificação , Staphylococcus/isolamento & purificação , Animais , Antibacterianos/farmacologia , Portador Sadio/microbiologia , Cães , Genes Bacterianos , Virilha/microbiologia , Testes de Sensibilidade Microbiana , Nigéria , Reação em Cadeia da Polimerase , Infecções Estafilocócicas/microbiologia , Staphylococcus/efeitos dos fármacos , Staphylococcus/enzimologiaRESUMO
Staphylococci are one of the most prevalent microorganisms in bovine mastitis. Staphylococcus spp. are widespread in the environment, and can infect animals and humans as opportunistic pathogens. The objective of this study was to determine the frequency of methicillin-resistance (MR) among coagulase-negative staphylococci (CoNS) previously obtained from milk of mastitic cows in Brazil and to characterize the antimicrobial resistance phenotype/genotype and the SCCmec type of MRCoNS isolates. Identification of MRCoNS was based on both biochemical and molecular methods. Susceptibility testing for eleven antimicrobials was performed by disk-diffusion agar. Antimicrobial resistance genes and SCCmec were investigated by specific PCRs. Twenty-six MRCoNS were detected (20 % of total CoNS), obtained from 24 animals, and were identified as follows: S. epidermidis (7 isolates), S. chromogenes (7), S. warneri (6), S. hyicus (5) and S. simulans (1). All MRCoNS isolates carried mecA while the mecC gene was not detected in any CoNS. The SCCmec IVa was demonstrated in nine MRCoNS, while the remaining 17 isolates harbored non-typeable SCCmec cassettes. In addition to oxacillin and cefoxitin resistance, MRCoNS showed resistance to tetracycline (n = 7), streptomycin (n = 6), tobramycin (n = 6), and gentamicin (n = 4), and harbored the genes tet(K) (n = 7), str (n = 3), ant(4') (n = 6) and aac(6')-aph(2â³) (n = 4), respectively. In addition, seven strains showed intermediate resistance to clindamycin and two to streptomycin, of which two harboured the lnu(B) and lsa(E) genes and two the aad(E) gene, respectively. One isolate presented intermediate erythromycin and clindamycin resistance and harbored an erm(C) gene with an uncommon 89-bp deletion rendering a premature stop codon. MRCoNS can be implicated in mastitis of cows and they constitute a reservoir of resistance genes that can be transferred to other pathogenic bacteria.
Assuntos
Coagulase/deficiência , Mastite Bovina/microbiologia , Resistência a Meticilina , Leite/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus/isolamento & purificação , Animais , Antibacterianos/farmacologia , Técnicas Bacteriológicas , Brasil , Bovinos , DNA Bacteriano/química , DNA Bacteriano/genética , Genes Bacterianos , Testes de Sensibilidade Microbiana , Técnicas de Diagnóstico Molecular , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Infecções Estafilocócicas/microbiologia , Staphylococcus/classificação , Staphylococcus/efeitos dos fármacos , Staphylococcus/genéticaRESUMO
The occurrence, resistance phenotype and molecular mechanisms of resistance of methicillin-resistant staphylococci from groin swabs of 109 clinically healthy dogs in Nsukka, Nigeria were investigated. The groin swab samples were cultured on mannitol salt agar supplemented with 10 µgof cloxacillin. Sixteen methicillin-resistant coagulase negative staphylococci (MRCoNS), all harbouring the mecA gene were isolated from 14 (12.8%) of the 109 dogs studied. The MRCoNS isolated were: S. sciuri subspecies rodentium, S. lentus, S. haemolyticus, and S. simulans with S. sciuri subspecies rodentium (62.5%) being the predominant species. Thirteen (81.3%) of the MRCoNS were resistant to tetracycline while 12 (75%) and 10 (62.5%) were resistant to kanamycin and trimthoprim-sulphamethoxazole respectively. None of the isolates was resistant to fusidic acid, linezolid and vancomycin. Thirteen (81.3%) of the MRCoNS were multi-drug resistance (MDR). Other antimicrobial genes detected were: blaZ, tet(K), tet(M), tet(L), erm(B), lnu(A), aacA-aphD, aphA3, str, dfr(G), cat pC221,and cat pC223. Methicillin-resistant staphylococci are common colonizers of healthy dogs in Nigeria with a major species detected being S. sciuri subsp. rodentium.
Assuntos
Animais , Cães , Portador Sadio/veterinária , Coagulase/deficiência , Resistência a Meticilina , Infecções Estafilocócicas/veterinária , Staphylococcus/classificação , Staphylococcus/isolamento & purificação , Antibacterianos/farmacologia , Portador Sadio/microbiologia , Genes Bacterianos , Virilha/microbiologia , Testes de Sensibilidade Microbiana , Nigéria , Reação em Cadeia da Polimerase , Infecções Estafilocócicas/microbiologia , Staphylococcus/efeitos dos fármacos , Staphylococcus/enzimologiaRESUMO
The occurrence, resistance phenotype and molecular mechanisms of resistance of methicillin-resistant staphylococci from groin swabs of 109 clinically healthy dogs in Nsukka, Nigeria were investigated. The groin swab samples were cultured on mannitol salt agar supplemented with 10 µgof cloxacillin. Sixteen methicillin-resistant coagulase negative staphylococci (MRCoNS), all harbouring the mecA gene were isolated from 14 (12.8%) of the 109 dogs studied. The MRCoNS isolated were: S. sciuri subspecies rodentium, S. lentus, S. haemolyticus, and S. simulans with S. sciuri subspecies rodentium (62.5%) being the predominant species. Thirteen (81.3%) of the MRCoNS were resistant to tetracycline while 12 (75%) and 10 (62.5%) were resistant to kanamycin and trimthoprim-sulphamethoxazole respectively. None of the isolates was resistant to fusidic acid, linezolid and vancomycin. Thirteen (81.3%) of the MRCoNS were multi-drug resistance (MDR). Other antimicrobial genes detected were: blaZ, tet(K), tet(M), tet(L), erm(B), lnu(A), aacA-aphD, aphA3, str, dfr(G), cat pC221,and cat pC223. Methicillin-resistant staphylococci are common colonizers of healthy dogs in Nigeria with a major species detected being S. sciuri subsp. rodentium.(AU)
Assuntos
Animais , Cães , Portador Sadio/veterinária , Coagulase/deficiência , Resistência a Meticilina , Infecções Estafilocócicas/veterinária , Staphylococcus/classificação , Staphylococcus/isolamento & purificação , Antibacterianos/farmacologia , Portador Sadio/microbiologia , Genes Bacterianos , Virilha/microbiologia , Testes de Sensibilidade Microbiana , Nigéria , Reação em Cadeia da Polimerase , Infecções Estafilocócicas/microbiologia , Staphylococcus/efeitos dos fármacos , Staphylococcus/enzimologiaRESUMO
Nasal carriage of methicillin-resistant coagulase-negative staphylococci (MR-CoNS) is highly prevalent in community subjects, but its dynamic has been little investigated. Nasal swabbing was performed in 2006 and 2008 in 154 Amerindians living isolated in French Guiana. MR-CoNS strains were identified and characterized by non-ß-lactam susceptibility testing and staphylococcal cassette chromosome mec element (SCCmec) typing, characterizing the associations of ccr and mec gene complex allotypes, and for MR Staphylococcus epidermidis (MRSE), multilocus variable number of tandem repeats analysis (MLVA) was used. The impact of sociodemographic and medical characteristics on the persistence of MR-CoNS carriage was assessed by bivariate analysis. Prevalence of MR-CoNS carriage was 50.6% in 2006 and 46.8% in 2008. The 274 MR-CoNS isolates, including S. epidermidis (n = 89, 62 MLVA patterns), Staphylococcus haemolyticus (n = 78), and Staphylococcus hominis (n = 72), exhibited 41 distinct ccr and mec gene complex associations. Persistent carriage (in 2006 and 2008), intermittent carriage (either in 2006 or 2008), and noncarriage were documented in 25.3, 47.4, and 27.3% of the participants, respectively. Persistent carriage of a given MRSE isolate was rarely observed (n = 8 isolates). Furthermore, no epidemiological factor, including antibiotic exposure, was associated with persistent carriage. The high diversity of MRSE clones and their ccr and mec gene complex associations contrasted with the high carriage rates in this isolated community, which might reflect the occurrence of SCCmec rearrangement and the generation of new MR-CoNS strains.
Assuntos
Coagulase/genética , Resistência a Meticilina/genética , Nariz/microbiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus epidermidis/genética , Staphylococcus haemolyticus/genética , Staphylococcus hominis/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/administração & dosagem , Antibacterianos/uso terapêutico , Técnicas de Tipagem Bacteriana , Portador Sadio , Cromossomos Bacterianos , Coagulase/deficiência , Feminino , Guiana Francesa/epidemiologia , Genes Bacterianos , Ligação Genética , Variação Genética , Humanos , Masculino , Resistência a Meticilina/efeitos dos fármacos , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Prevalência , Infecções Estafilocócicas/tratamento farmacológico , Infecções Estafilocócicas/epidemiologia , Staphylococcus epidermidis/efeitos dos fármacos , Staphylococcus epidermidis/isolamento & purificação , Staphylococcus haemolyticus/efeitos dos fármacos , Staphylococcus haemolyticus/isolamento & purificação , Staphylococcus hominis/efeitos dos fármacos , Staphylococcus hominis/isolamento & purificaçãoRESUMO
In the present study, 188 coagulase-negative Staphylococcus (CNS) strains were isolated from bovine mastitis cases from 56 different Brazilian dairy herds, located in the Southeast region of the country, and were tested for antimicrobial substance production. Twelve CNS strains (6.4%) exhibited antagonistic activity against a Corynebacterium fimi indicator strain. Most antimicrobial substances were sensitive to proteolytic enzymes suggesting that they might be bacteriocins (Bac). Amongst the CNS producers, six were identified as S. epidermidis, two as S. simulans, two as S. saprophyticus, one as S. hominis and one as S. arlettae. Plasmid profile analysis of these strains revealed the presence of at least one plasmid. The Bac(+) strains presented either no or few antibiotic resistance phenotypes. Three strains were shown to produce a bacteriocin either identical or similar to aureocin A70, a bacteriocin previously isolated from an S. aureus strain isolated from food. The remaining Bac(+) strains produce antimicrobial peptides that seem to be distinct from the best characterised staphylococcal bacteriocins described so far. Some of them were able to inhibit Listeria monocytogenes, an important food-borne pathogen, and several strains of Streptococcus agalactiae associated with bovine mastitis, suggesting a potential use of these bacteriocins either in the prevention or in the treatment of streptococcal mastitis.
Assuntos
Bacteriocinas/biossíntese , Mastite Bovina/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus/metabolismo , Animais , Antibacterianos/farmacologia , Bacteriocinas/metabolismo , Southern Blotting/veterinária , Bovinos , Coagulase/deficiência , DNA Bacteriano/química , DNA Bacteriano/genética , Farmacorresistência Bacteriana , Feminino , Listeria monocytogenes/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana/veterinária , Peptídeo Hidrolases/metabolismo , Reação em Cadeia da Polimerase/veterinária , Infecções Estafilocócicas/metabolismo , Infecções Estafilocócicas/microbiologia , Staphylococcus/enzimologia , Staphylococcus/genética , Staphylococcus/crescimento & desenvolvimentoRESUMO
A working scheme developed in our laboratory for identification (by species group and species) of coagulase-negative staphylococci (CNS) was evaluated with 201 consecutive isolates and then validated by using the reference method of Kloos and Schleifer (W. E. Kloos and K. H. Schleifer, J. Clin. Microbiol. 1:82-88, 1975). This five-test simple scheme (referred to here as the simple scheme) combines the novobiocin susceptibility test with tests for urease, pyrrolidonyl arylamidase, ornithine decarboxylase, and aerobic acid from mannose. The addition of one or two tests within a particular species group could then positively identify the isolate. Two commercial systems, Staph-Zym (Rosco) and API-Staph (bioMérieux), along with results obtained by using Rosco diagnostic tablets (nongrowth tests), were also compared with the reference method. One isolate could not be identified even by the reference method. Of the remaining 200 strains, 191 (95.5%) strains were correctly identified with Staph-Zym and 171 strains (85.5%) were correctly identified with API-Staph. The most frequent clinical CNS species isolated were Staphylococcus epidermidis (50.5%), S. haemolyticus (18.5%), S. saprophyticus subsp. saprophyticus (16.0%), S. lugdunensis (6.0%), and S. warneri (2.5%). The simple scheme validated with the reference method has demonstrated an excellent correlation in the identification of the three most frequent species isolated: S. epidermidis, S. haemolyticus, and S. saprophyticus subsp. saprophyticus. With the simple scheme, identification of CNS was possible within 24 h after the enzymatic tests were used, whereas up to 72 h is necessary for the growth tests. This methodology would be very useful in any clinical microbiology laboratory for the presumptive identification of CNS species groups and species.