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1.
Rev. Soc. Bras. Med. Trop ; 52: e20190095, 2019. tab, graf
Article in English | LILACS | ID: biblio-1013299

ABSTRACT

Abstract INTRODUCTION Staphylococcus aureus is a major nosocomial pathogen that is associated with high virulence and the rapid development of drug resistance. METHODS We analyzed and compared the antimicrobial resistance, virulence profiles, and molecular epidemiology of 67 S. aureus strains, including 36 methicillin-sensitive (MSSA) and 31 methicillin-resistant (MRSA) strains recovered from a public hospital located in south-eastern Brazil. RESULTS The clones circulating in this hospital presented a great diversity, and the majority of the strains were related to clones responsible for causing worldwide epidemics: these included USA100 (New York/Japan clone), USA300, and USA600. The 31 MRSA (22 SCCmecII and 9 SCCmecIV) and 36 MSSA strains exhibited low resistance against gentamicin and trimethoprim/sulfamethoxazole. No MRSA strain showed resistance to tetracycline. Virulence gene carriage was more diverse and abundant in MSSA than in MRSA. Of the evaluated adhesion-related genes, ebpS was the most prevalent in both MSSA and MRSA strains. The genes bbp and cna showed a strong association with MSSA strains. CONCLUSIONS Our findings reinforce the idea that MSSA and MRSA strains should be carefully monitored, owing to their high pathogenic potential.


Subject(s)
Humans , Staphylococcus aureus/drug effects , Staphylococcus aureus/genetics , Methicillin Resistance , Virulence Factors/genetics , Methicillin-Resistant Staphylococcus aureus/genetics , Anti-Bacterial Agents/pharmacology , Staphylococcal Infections/epidemiology , Staphylococcus aureus/isolation & purification , Staphylococcus aureus/classification , Virulence/genetics , Brazil/epidemiology , Microbial Sensitivity Tests , Molecular Epidemiology , Tertiary Care Centers , Hospitals, Public
2.
Pesqui. vet. bras ; 38(7): 1358-1364, July 2018. tab, ilus
Article in English | LILACS, VETINDEX | ID: biblio-976437

ABSTRACT

This study evalueted the prevalence of Staphylococcus aureus, Streptococcus agalactiae and Escherichia coli in milk samples from 257 goats (513 half-udders) and ten bulk tanks, from ten dairy goat farms of São Paulo State, Brazil, by multiplex-PCR. The samples were screened by microbiological culture (gold-standard), and tested by different multiplex-PCR protocols for the detection of each bacterium. A total of 178 half-udders resulted positive by microbiological culture, with coagulase-negative staphylococci (70%), S. aureus (13.5%), S. intermedius (7.9%), and Enterobacteriaceae (4%) the prevalent pathogens. In other way, multiplex-PCR detected 173 pathogens in 151/523 (28.9%; CI95% 25.2-32.9%) milk samples 144/513 (28.1%) half-udders and 7/10 (70%) bulk tanks, with E. coli (86/162, 51.9%) and S. aureus (50/162, 30.9%) the prevalent ones in half-udders, and S. aureus (6/10, 60%) and E. coli (4/5, 36.4%) in bulk tanks. Multiplex-PCR showed a high performance for the detection of three bacteria at a time in mastitic goat milk direct from half-udders or bulk tanks. Thus, this multiplex-PCR protocol proved to be an adequate tool for the identification of the most common mastitis pathogens, independent of their phenotypic characteristics in the diagnosis of clinical mastitis in goats, allowing a continuous and better vigilance and monitoring the herd, being included in quality programs.(AU)


Este estudo avaliou por multiplex-PCR a prevalência de Staphylococcus aureus, Streptococcus agalactiae e Escherichia coli em amostras de leite de 257 caprinos (513 tetos) e dez tanques de expansão, em dez fazendas leiteiras do estado de São Paulo, Brasil. As amostras foram triadas por cultura microbiológica (padrão-uro) e testadas por diferentes protocolos multiplex-PCR para a detecção de cada bactéria. Um total de 178 amostras de leite foram positivos na cultura microbiológica, com estafilococos coagulase-negativos (70%), S. aureus (13,5%), S. intermedius (7,9%) e Enterobacteriaceae (4%) como patógenos prevalentes. Por outro lado, a PCR multiplex detectou 173 patógenos em 151/523 (28,9%, IC95% 25,2-32,9%) amostras de leite, 144/513 (28,1%) amostras de tetos e 7/10 (70%) em tanques de expansão, E. coli (86/162, 51,9%) e S. aureus (50/162, 30,9%) foram identificados nas amostras de tetos e S. aureus (6/10, 60%) e E. coli (4/5, 36,4%) em tanques expansão. Multiplex-PCR mostrou um alto desempenho para a detecção das três bactérias em leite de cabra com mastite ou em tanques de expansão. Dessa forma, este protocolo multiplex-PCR provou ser uma ferramenta adequada para a identificação dos patógenos mais comuns da mastite, independentemente de suas características fenotípicas no diagnóstico de mastite clínica em caprinos, permitindo uma vigilância contínua e melhor acompanhamento do rebanho, sendo incluído em programas de qualidade.(AU)


Subject(s)
Animals , Staphylococcus aureus/classification , Streptococcus agalactiae/classification , Ruminants/abnormalities , Escherichia coli/classification
3.
São Paulo; s.n; s.n; 2018. 114 p. graf, tab, ilus.
Thesis in Portuguese | LILACS | ID: biblio-995936

ABSTRACT

Pacientes com queimaduras e outras lesões cutâneas de grande extensão apresentam alta propensão a infecções multirresistentes. Neste contexto, o objetivo deste trabalho é a obtenção de nanopartículas de quitosana carregadas com composto derivado de 5-nitro-heterocíclico com estrutura análoga à nifuroxazida, N'-((5-nitrofurano-2-il)metileno)-2-benzidrazida (C-H), que apresentou importante atividade frente a diversas cepas de bactérias multirresistentes. Por sua vez, a quitosana é um biopolímero com atividade antimicrobiana, analgésica, regeneradora tecidual e que, mediante contato com exsudato de lesões cutâneas forma filmes hidrogéis protetores no local de aplicação, sendo estas atividades importantes para a prevenção e tratamento de infecções e da exsudação excessiva de lesões de grande extensão. As nanopartículas de quitosana carregadas com o composto (Nps-H) foram obtidas pelo método de gelificação iônica com tripolifosfato de sódio como agente reticulante, variando a concentração de NaCl e polissorbato 80 do sistema, orientada por análise fatorial. As Nps-H obtidas foram caracterizadas por Dynamic Light Scattering (DLS) para determinação do tamanho, índice de polidispersão (IPD) e potencial zeta (ζ). A eficiência de encapsulação (EE%) foi determinada por espectrofotometria UV/VIS a 370 nm por método indireto. Entre os experimentos desenvolvidos, aquele que apresentou os melhores resultados resultou em partículas de tamanho médio de 321 d.nm, IPD 0,18, Pζ +37 mV e EE% de 48%. A morfologia e superfície das Nps-H foram analisadas por Microscopia Eletrônica de Varredura (MEV) e mostraram que as Nps-H são esféricas e de superfície irregular. A partir da obtenção e caracterização das Nps-H determinou-se a concentração inibitória mínima (CIM) das Nps-H, do composto livre (C-H) e das nanopartículas de quitosana vazias (Nps-Cs) por método colorimétrico e microdiluição frente às cepas de Staphylococcus aureus ATCC 29213, hVISA e ORSA. As Nps-H apresentaram atividade bastante superior comparando-se ao C-H e as Nps-Cs frente às cepas de S. aureus estudadas. Com vista à preparação de uma formulação farmacêutica estável, partiu-se para a liofilização das Nps-H e com esse objetivo realizou-se análises térmicas das Nps-H por Differential Scanning Calorimetric (DSC) para determinação das temperaturas de transição vítrea e eutética (Tg' e Teut) e análise por criomicroscopia para determinação da temperatura de colapso (Tcol). Amostras de Nps-H com lio/crioprotetores glicina, lactose e sacarose a diferentes concentrações foram liofilizadas a -40 ºC a 100 mTorr. As amostras de Nps-H com lactose e sacarose ambas a 2,5% e 5% demonstraram preservar as características originais das Nps-H após o processo de liofilização. Observou-se, com os resultados obtidos neste trabalho que as Nps-H representam uma promissora alternativa na prevenção e tratamento de pacientes com lesões cutâneas infeccionadas por bactérias multirresistentes e no controle da exsudação excessiva, principalmente por suas atividades terapêuticas em conjunto, diminuindo a mortalidade e morbidade desses quadros


Patients with burns and others extensive skin lesions show high propension to multiresistant infections. In this context, the aim of this project is to obtain chitosan nanoparticles carried with 5-nitro-heterocyclic derivate with structure analogue to nifuroxazide N'-((5-nitrofuran-2-yl) methylene)-2-benzhydrazide, that showed important activity against multiresistant bacteria strains. In its turn, the chitosan is a biopolymer with antimicrobial, analgesic, tecidual regenerator activities and by contact with excessive burns and extensive skin lesions exsudate process, form protective hydrogel films on place of application, being these activities important to infection and excessive exsudate treatment and prevention of extensive burns and lesions. The chitosan nanoparticles carried with compound (Nps-H) were obtained by ionic gelation method with sodium tripolyphosphate as crosslinker agent, varing the NaCl and polysorbate 80 concentrations in the system, oriented by factorial analyze. The Nps-H obtained were characterized by Dynamic Light Scattering (DLS) to size determination, polydispersion index (PDI) and zeta potential (ζ-P). The encapsulation efficiency (EE%) were determined by spectrophotometry UV/VIS at 370 nm by indirect method. Bepolissorbato the experiments developed, the one who showed the best results, resulted in particles with size of 321 d.nm, PDI of 0,18, ζ-P of +37 mV and EE% of 48%. The Nps-H morphology and surface were analyzed by Scanning Electronic Microscopy (SEM) and showed that Nps-H are spherical and with irregular surface. Starting of Nps-H obtain and characterization were determined the Nps-H, free compound (C-H) and empty chitosan nanoparticles (Nps-Cs) Minimal Inhibitory Concentrations (MIC) by colorimetric method and microdilution against Staphylococcus aureus ATCC 29213, ORSA and hVISA strains. The Nps-H showed the superior activity comparing to C-H and Nps-H against all strains tested. With a view to preparation of stable pharmaceutic formulation, started to Nps-H freeze-drying and with this aim, were realized Nps-H thermal analyzes by Differential Scanning Calorimetric (DSC) to determine glass transition and euthetic temperatures (Tg' and Teut) and the cryomicroscopy analyze to determine collapse temperature (Tcol). The Nps-H samples with lyo/cryoprotectants as glycine, lactose and sucrose at different concentrations were lyophilized at -40 ºC at 100 mTorr. The Nps-H samples with lactose and sucrose both at 2,5% and 5% demonstrated to preserve the original Nps-H characteristics after freeze-drying process. Were observed, with the results obtained in this project that Nps-H represent the promising alternative to prevention and treatment of patients with infected skin lesions by multiresitant bacteria and to control of excessive exudate process, mainly by their therapeutic activities combined, decreasing the mortality and morbidity of these cases


Subject(s)
Chitosan/analysis , Nanoparticles/classification , Anti-Infective Agents/analysis , Skin , Staphylococcus aureus/classification , Burns , Drug Discovery
4.
Braz. j. infect. dis ; 20(6): 525-533, Nov.-Dec. 2016. tab, graf
Article in English | LILACS | ID: biblio-828162

ABSTRACT

ABSTRACT The spread of pandemic Staphylococcus aureus clones, mainly methicillin-resistant S. aureus (MRSA), must be kept under surveillance to assemble an accurate, local epidemiological analysis. In Ecuador, the prevalence of the USA300 Latin American variant clone (USA300-LV) is well known; however, there is little information about other circulating clones. The aim of this work was to identify the sequence types (ST) using a Multiple-Locus Variable number tandem repeat Analysis 14-locus genotyping approach. We analyzed 132 S. aureus strains that were recovered from 2005 to 2013 and isolated in several clinical settings in Quito, Ecuador. MRSA isolates composed 46.97% (62/132) of the study population. Within MRSA, 37 isolates were related to the USA300-LV clone (ST8-MRSA-IV, Panton-Valentine Leukocidin [PVL] +) and 10 were related to the Brazilian clone (ST239-MRSA-III, PVL−). Additionally, two isolates (ST5-MRSA-II, PVL−) were related to the New York/Japan clone. One isolate was related to the Pediatric clone (ST5-MRSA-IV, PVL−), one isolate (ST45-MRSA-II, PVL−) was related to the USA600 clone, and one (ST22-MRSA-IV, PVL−) was related to the epidemic UK-EMRSA-15 clone. Moreover, the most prevalent MSSA sequence types were ST8 (11 isolates), ST45 (8 isolates), ST30 (8 isolates), ST5 (7 isolates) and ST22 (6 isolates). Additionally, we found one isolate that was related to the livestock associated S. aureus clone ST398. We conclude that in addition to the high prevalence of clone LV-ST8-MRSA-IV, other epidemic clones are circulating in Quito, such as the Brazilian, Pediatric and New York/Japan clones. The USA600 and UK-EMRSA-15 clones, which were not previously described in Ecuador, were also found. Moreover, we found evidence of the presence of the livestock associated clone ST398 in a hospital environment.


Subject(s)
Humans , Male , Female , Infant, Newborn , Infant , Child, Preschool , Child , Adolescent , Adult , Middle Aged , Aged , Aged, 80 and over , Young Adult , Staphylococcal Infections/microbiology , Staphylococcus aureus/genetics , Bacterial Toxins/genetics , Exotoxins/genetics , Leukocidins/genetics , Anti-Bacterial Agents/pharmacology , Staphylococcus aureus/isolation & purification , Staphylococcus aureus/classification , DNA, Bacterial , Microbial Sensitivity Tests , Prevalence , Virulence Factors/genetics , Ecuador , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Methicillin-Resistant Staphylococcus aureus/classification , Methicillin-Resistant Staphylococcus aureus/genetics , Multilocus Sequence Typing , Genotype
5.
Braz. j. microbiol ; 46(2): 535-543, Apr-Jun/2015. tab, graf
Article in English | LILACS | ID: lil-749725

ABSTRACT

The objective of this study was to characterize the clonal profile, virulence factors and antimicrobial resistance, particularly oxacillin resistance, of Staphylococcus aureus isolated from sheep milk. Milk samples were collected from all teats for the California Mastitis Test (CMT), somatic cell count, identification of S. aureus, investigation in these strains of genes encoding toxins (sea, seb, sec, sed, tst), biofilm (icaA, icaC, icaD, bap), leukocidin (luk-PV) oxacillin resistance by mecA gene detection and susceptibility testing (12 antibiotics). Messenger RNA expression was evaluated by RT-PCR in isolates carrying toxin and biofilm genes. Biofilm formation was also evaluated phenotypically by adherence to polystyrene plates. The clonal profile of S. aureus was investigated by pulsed-field gel electrophoresis. A total of 473 milk samples were collected from 242 animals on three farms and 20 S. aureus strains were isolated and none carried the mecA gene. The two sec gene-positive isolates and the isolates carrying the tst and luk-PV genes were positive by RT-PCR. Staphylococcus aureus isolated from the three flocks studied showed high susceptibility to the drugs tested and none was biofilm producer, indicating that biofilm formation was not a virulence factor causing infection by these strains. The typing of 17 S. aureus isolates revealed the presence of a common clone on the three farms studied, and the presence and expression of the sec and tst genes in one strain of this clone suggest the possible acquisition of virulence genes by this clone, a fact that is important for animal health and food hygiene.


Subject(s)
Animals , Milk/microbiology , Staphylococcus aureus/isolation & purification , Staphylococcus aureus/physiology , Anti-Bacterial Agents/pharmacology , Biofilms/growth & development , California , Electrophoresis, Gel, Pulsed-Field , Gene Expression Profiling , Microbial Sensitivity Tests , Molecular Typing , Reverse Transcriptase Polymerase Chain Reaction , Sheep , Staphylococcus aureus/classification , Staphylococcus aureus/drug effects , Virulence Factors/genetics
6.
Rev. bras. plantas med ; 17(4,supl.2): 865-874, 2015. tab, graf
Article in English | LILACS | ID: lil-771170

ABSTRACT

ABSTRACT The extracts and fractions of leaves and branches of Protium hebetatum D. C. Daly (Burseraceae) were investigated for their antibacterial activity and chemical composition. The methanol extract of branches (EMG) was considered active against the Escherichia coli and the Proteus vulgaris, showing an inhibition zone of 13 mm, and was selected for bioassay-guided phytochemical fractionation. From the technique of broth microdilution, the extract was considered a moderate inhibitor against Staphylococcus aureus, Pseudomonas aeruginosa and Enterococcus faecalis, with a minimum inhibitory concentration (MIC) of 1 mg/mL. The dichloromethane fraction was considered a moderate inhibitor against S. aureus (MIC of 1 mg/mL) and a potent inhibitor against E. faecalis (MIC of 0.5 mg/mL). F1, F2, F5 and F6 from chromatographic column of dichloromethane fraction were considered moderate inhibitors against S. aureus (MIC of 1 mg/mL). Through analysis by a gas chromatography mass spectrometry, eighteen compounds were identified, from which thirteen (isoeugenol, p-vinylguaiacol, metoxyeugenol, coumarin, 5-hydroxy-scopoletin, 4,7-dihydroxy-6-metoxicromam-2-one, 4[(1E]-3-hydroxy-1-propenyl)-2-methoxyphenol, piperonal, scoparon, o-guaiacol, spathulenol, seringol and antiarol) are unprecedented in these species. We also identified the triterpenes α-amyrin and β-amyrin, the steroids stigmasterol and sitosterol and the coumarin scopoletin, which was closely linked to the antibacterial activity of the samples.


RESUMO Atividade antibacteriana e compostos químicos de folhas e galhos de Protium hebetatum. Extratos e frações de folhas e galhos de Protium hebetatum D. C. Daly (Burseraceae) foram investigados quanto sua atividade antibacteriana e composição química. O extrato metanólico dos galhos (EMG) foi considerado ativo contra Escherichia coli e Proteus vulgaris, apresentando um halo de inibição de 13 mm, sendo selecionado para um fracionamento fitoquímico biomonitorado. A partir da técnica de microdiluição em caldo o EMG foi considerado um inibidor moderado contra Staphylococcus aureus, Pseudomonas aeruginosa e Enterococcus faecalis, apresentando uma concentração inibitória mínima (CIM) de 1mg/mL. A fração diclorometânica foi considerada inibidora moderada contra S. aureus (CIM de 1 mg/mL) e inibidora potente contra E. faecalis (CIM de 0,5 mg/mL). F1, F2, F5 e F6 provenientes da fração diclorometânica foram consideradas inibidoras moderadas contra S. aureus (CIM de 1 mg/mL). Através da análise por cromatografia gasosa acoplada a espectrometria de massa, foram identificados dezoitos compostos, dos quais treze (isoeugenol, p-vinilguaiacol, metoxieugenol, cumarina, 5-hidroxi-escopoletina, 4,7-dihidroxi-6-metoxicromam-2-ona, 4[(1E]-3-hidroxi-1-propenil)-2-methoxifenol, piperonal, escoparona, o-guaiacol, espatulenol, seringol e antiarol) foram identificados pela primeira vez nesta espécie. Foram também identificados os triterpenos α-amirina e β-amirina, os esteroides estigmasterol e sitosterol e a cumarina escopoletina, que estão intimamente ligados à atividade antibacteriana da espécie.


Subject(s)
Anti-Infective Agents/classification , Burseraceae/classification , Chemical Compounds/analysis , Coumarins/pharmacology , Enterococcus faecalis/classification , Staphylococcus aureus/classification
7.
Braz. j. pharm. sci ; 50(3): 567-572, Jul-Sep/2014. tab, graf
Article in English | LILACS | ID: lil-728703

ABSTRACT

Vancomycin (VAN) is the gold standard therapy for Methicillin-resistant Staphylococcus aureus (MRSA) infections such as bacteremia and endocarditis. However, VAN suboptimal dosing for serious infections caused by S. aureus isolates that have elevated minimum inhibitory concentration (MIC), could be associated with poor outcome. Better understanding of VAN pharmacokinetics and pharmacodynamics (PK/PD) has led to the creation of new recommendations with optimized dosing regimens for the treatment of MRSA infections. For severe infectious, such as pneumonia and endocarditis, a VAN serum trough concentration of 15-20 mg/L at the steady state should be targeted. The aim of this study was to show how a nomogram with updated VAN dosing was devised and how it was implemented in the electronic prescribing (e-prescribing) system of a teaching hospital. VAN loading dose and maintenance doses were calculated from a pharmacokinetic equation using basic parameters: weight, estimated creatinine clearance, as well as peak and trough serum concentrations. The implementation of the VAN dosing nomogram in the hospital e-prescribing system definitively changed the long-standing medical prescription fallacy of "same dose fits all". Finally, this computer-based electronic program has allowed a wide-ranging intervention and should be recognized as a powerful tool for implementation in antimicrobial stewardship programs.


Vancomicina (VAN) é utilizada como primeira escolha na terapia de infecções causadas por Staphylococcus aureus resistentes à meticilina (MRSA), como bacteremia e endocardite. Entretanto, o aumento na concentração inibitória mínima (CIM) de isolados de S. aureus e doses subterapêuticas de VAN podem estar associados à falha terapêutica. Para o melhor entendimento sobre o perfil farmacocinético e farmacodinâmico (PK/PD) da VAN foram elaboradas novas recomendações para terapia de infecções causadas por MRSA. Para terapia de infecções graves, como pneumonia e endocardite, a concentração sérica do vale de VAN de 15-20 mg/L no estado de equilíbrio dinâmico deve ser o alvo. O objetivo do estudo foi desenvolver um nomograma com doses atualizadas de VAN e demonstrar como ele foi implementado no sistema de prescrição eletrônica em um Hospital Universitário. As doses de ataque e manutenção foram calculadas a partir de equações farmacocinéticas, utilizando parâmetros fundamentais: peso, depuração de creatinina, concentrações séricas do pico e do vale. A implementação de um nomograma de doses de VAN em um sistema de prescrição eletrônica modificou definitivamente o inadequado hábito de que "a mesma dose cabe em todos". Finalmente, esta abrangente ferramenta tecnológica deve ser considerada como uma robusta estratégia num programa de uso racional de antibióticos.


Subject(s)
Anti-Bacterial Agents , Electronic Prescribing/classification , Nomograms , Vancomycin/pharmacokinetics , Methicillin/pharmacokinetics , Staphylococcus aureus/classification
8.
Braz. j. microbiol ; 45(3): 857-859, July-Sept. 2014. graf
Article in English | LILACS | ID: lil-727014

ABSTRACT

This study evaluated the relationship between previous colonization of the oropharynx and development of ventilator-associated pneumonia through the classification of genomic fingerprint pattern by pulsed-field gel electrophoresis of both oxacillin-resistant and oxacillin-susceptible Staphylococcus aureus isolates obtained from hospitalized patients in an intensive care unit.


Subject(s)
Humans , Carrier State/microbiology , Oropharynx/microbiology , Pneumonia, Staphylococcal/epidemiology , Pneumonia, Ventilator-Associated/epidemiology , Genotype , Molecular Epidemiology , Molecular Typing , Pneumonia, Staphylococcal/microbiology , Pneumonia, Ventilator-Associated/microbiology , Staphylococcus aureus/classification , Staphylococcus aureus/genetics , Staphylococcus aureus/isolation & purification
9.
Braz. j. microbiol ; 44(4): 1155-1161, Oct.-Dec. 2013. graf, tab
Article in English | LILACS | ID: lil-705275

ABSTRACT

An increase in the consumption of fruit juices and minimally processed fruits salads has been observed in recent years all over the world. In this work, the microbiological quality of artisan fruit salads was analysed. Faecal coliforms, Salmonella spp, Shigella spp, Yersinia enterocolitica and Escherichia coli O157:H7 were not detected; nevertheless, eleven strains of Staphylococcus aureus were isolated. By multiplex PCR, all isolates showed positive results for S. aureus 16S rRNA gene and 63.6% of them were positive for sea gene. Furthermore, PCR sea positive strains were able to produce the corresponding enterotoxin. Finally, the inactivation of these strains in fruit salads by nisin, lysozyme and EDTA, was studied. EDTA produced a total S. aureus growth inhibition after 60 h of incubation at a concentration of 250 mg/L. The presence of S. aureus might indicate inadequate hygiene conditions during salad elaboration; however, the enterotoxigenicity of the strains isolated in this study, highlights the risk of consumers' intoxication. EDTA could be used to inhibit the growth of S. aureus in artisan fruit salads and extend the shelf life of these products.


Subject(s)
Enterotoxins/genetics , Fruit/microbiology , Staphylococcus aureus/isolation & purification , Argentina , DNA, Bacterial/genetics , DNA, Ribosomal/genetics , Multiplex Polymerase Chain Reaction , /genetics , Staphylococcus aureus/classification , Staphylococcus aureus/genetics , Staphylococcus aureus/metabolism
10.
Braz. j. pharm. sci ; 49(1): 13-27, Jan.-Mar. 2013. ilus, tab
Article in English | LILACS | ID: lil-671397

ABSTRACT

Osteomyelitis is an inflammatory bone disorder caused by infection, leading to necrosis and destruction of bone. It can affect all ages, involve any bone, become a chronic disease and cause persistent morbidity. Treatment of osteomyelitis is challenging particularly when complex multiresistant bacterial biofilm has already been established. Bacteria in biofilm persist in a low metabolic phase, causing persistent infection due to increased resistance to antibiotics. Staphylococcus aureus and Staphylococcus epidermidis are the most common causative organism responsible for more than 50% of osteomyelitis cases. Osteomyelitis treatment implies the administration of high doses of antibiotics (AB) by means of endovenous and oral routes and should take a period of at least 6 weeks. Local drug delivery systems, using non-biodegradable (polymethylmethacrylate) or biodegradable and osteoactive materials such as calcium orthophosphates bone cements, have been shown to be promising alternatives for the treatment of osteomyelitis. These systems allow the local delivery of AB in situ with bactericidal concentrations for long periods of time and without the toxicity associated with other means of administration. This review examines the most recent literature evidence on the causes, pathogeneses and pharmacological treatment of osteomyelitis. The study methodology consisted of a literature review in Google Scholar, Science Direct, Pubmed, Springer link, B-on. Papers from 1979 till present were reviewed and evaluated.


A osteomielite é um processo inflamatório do tecido ósseo, de origem infecciosa, que resulta em destruição inflamatória, necrose e formação de novo osso. Pode aparecer em qualquer idade, afetar qualquer osso e tornar-se uma doença crônica com morbidade persistente. Apesar dos progressos na quimioterapia infecciosa, o tratamento da osteomielite é caro e difícil, em particular quando associada à presença de biofilmes bacterianos, especialmente de Staphylococcus aureus e Staphylococcus epidermidis. O tratamento da osteomielite inclui a administração de doses elevadas de antibióticos (AB) por via endovenosa e oral, durante um período de pelo menos 6 semanas. Os sistemas de veiculação localizada de fármacos, utilizando materiais não biodegradáveis (polimetilmetacrilato) ou biodegradáveis e osteoativos como os cimentos ósseos de ortofosfatos de cálcio e vidro bioativo, surgiram como uma alternativa promissora para o tratamento da osteomielite. Estes sistemas permitem a veiculação de AB in situ com concentrações bactericidas por longos períodos de tempo e sem a toxicidade associada às outras vias de administração. O presente trabalho propõe uma revisão da literatura relativa às causas, à patogenia e ao tratamento farmacológico da osteomielite. A metodologia do estudo da revisão consistiu numa pesquisa bibliográfica, nas bases de dados Google Scholar, Science Direct, Pubmed, Springer link, B-on. Foram revistos e analisados diversos artigos publicados desde o ano de 1979.


Subject(s)
Osteomyelitis/classification , Osteomyelitis/diagnosis , Anti-Bacterial Agents , Osteomyelitis/pathology , Staphylococcus aureus/classification
11.
Braz. j. microbiol ; 44(2): 493-498, 2013. tab
Article in English | LILACS | ID: lil-688584

ABSTRACT

Staphylococcus aureus is one of the most frequent mastitis causative agents in small ruminants. The expression of most virulence genes of S. aureus is controlled by an accessory gene regulator (agr)locus. This study aimed to ascertain the prevalence of the different agr groups and to evaluate the occurrence of encoding genes for cytotoxin, adhesins and toxins with superantigen activity in S. aureus isolates from milk of ewes with clinical and subclinical mastitis in sheep flocks raised for meat production The agr groups I and II were identified in both cases of clinical and subclinical mastitis. Neither the arg groups III and IV nor negative agr were found. The presence of cflA gene was identified in 100% of the isolates. The frequency of hla and lukE-D genes was high -77.3 and 82.8%, respectively and all isolates from clinical mastitis presented these genes. The sec gene, either associated to tst gene or not, was identified only in isolates from subclinical mastitis. None of the following genes were identified: bbp, ebpS, cna, fnbB, icaA, icaD, bap, hlg, lukM-lukF-PV and se-a-b-d-e.


Subject(s)
Animals , Bacterial Proteins/genetics , Carrier State/veterinary , Mastitis/veterinary , Sheep Diseases/microbiology , Staphylococcal Infections/veterinary , Staphylococcus aureus/genetics , Trans-Activators/genetics , Virulence Factors/genetics , Brazil , Carrier State/microbiology , Genes , Genotype , Mastitis/microbiology , Milk/microbiology , Sheep , Staphylococcal Infections/microbiology , Staphylococcus aureus/classification , Staphylococcus aureus/isolation & purification
12.
Rev. Soc. Bras. Med. Trop ; 45(6): 717-722, Nov.-Dec. 2012. tab
Article in English | LILACS | ID: lil-661073

ABSTRACT

INTRODUCTION: In venous ulcers, the presence of Staphylococcus aureus and coagulase-negative staphylococcus resistance phenotypes can aggravate and limit the choices for treatment. METHODS: Staphylococcus isolated from 69 patients (98 ulcers) between October of 2009 and October of 2010 were tested. The macrolide, lincosamide, streptogramin B (MLS B) group resistance phenotype detection was performed using the D-test. Isolates resistant to cefoxitin and/or oxacillin (disk-diffusion) were subjected to the confirmatory test to detect minimum inhibitory concentration (MIC), using oxacillin strips (E-test®). RESULTS: The prevalence of S. aureus was 83%, and 15% of coagulase-negative staphylococcus (CoNS). In addition were detected 28% of methicillin-resistant Staphylococcus aureus (MRSA) and 47% of methicillin-resistant coagulase-negative staphylococcus (MRCoNS). Among the S. aureus, 69.6% were resistant to erythromycin, 69.6% to clindamycin, 69.6% to gentamicin, and 100% to ciprofloxacin. Considering the MRSA, 74% were highly resistant to oxacillin, MIC ≥ 256µg/mL, and the MLS Bc constitutive resistance predominated in 65.2%. Among the 20 isolates sensitive to clindamycin, 12 presented an inducible MLS B phenotype. Of the MRCoNS, 71.4%were resistant to erythromycin, ciprofloxacin and gentamicin. Considering the isolates positive for β-lactamases, the MIC breakpoint was between 0.5 and 2µg/mL. CONCLUSIONS: The results point to a high occurrence of multi-drug resistant bacteria in venous ulcers in primary healthcare patients, thus evidencing the need for preventive measures to avoid outbreaks caused by multi-drug resistant pathogens, and the importance of healthcare professionals being able to identifying colonized versus infected venous ulcers as an essential criteria to implementing systemic antibacterial therapy.


INTRODUÇÃO: Em úlceras venosas, a presença de Staphylococcus aureus e coagulase negativo com fenótipos de resistência pode constituir fator agravante e limita as opções terapêuticas. MÉTODOS: Foram avaliados estafilococos isolados de 69 pacientes, representando 98 úlceras no período de outubro de 2009 a outubro de 2010. A detecção fenotípica da resistência ao grupo macrolide, lincosamide, streptogramin B (MLS B) foi realizada pelo D-test. Isolados resistentes a cefoxitina e/ou oxacilina (disco-difusão) foram submetidos ao teste confirmatório para detecção da minimum inhibitory concentration (MIC), empregando fitas de oxacilina (E-test®). RESULTADOS: A prevalência de S. aureus foi de 83% e de 15% de coagulase-negative staphylococcus (CoNS). Identificou-se 28% de methicillin-resistant Staphylococcus aureus (MRSA) e 47% de methicillin-resistant coagulase-negative staphylococcus (MRCoNS). Entre o S. aureus, 69,6% apresentaram resistência a eritromicina, 69,6% a clindamicina, 69,6% a gentamicina e 100% a ciprofloxacina. Setenta e quatro por cento dos MRSA apresentaram elevado nível de resistência a oxacilina, MIC ≥ 256µg/mL, e em 65,2% predominou a resistência constitutiva MLS Bc. Dos 20 isolados sensíveis a clindamicina, 12 apresentaram fenótipo MLS B induzível. Um total de 71,4% dos MRCoNS apresentaram resistência a eritromicina, ciprofloxacina e gentamicina. Dos isolados positivos para a enzima β-lactamases, as MIC tiveram breakpoint entre 0,5 a 2µg/mL. CONCLUSÕES: Os resultados sinalizam elevada ocorrência de bactérias multirresistentes em úlceras venosas de pacientes recebendo atenção primária, evidenciando a necessidade de medidas preventivas que evitem surtos causados por patógenos resistentes a múltiplas drogas e a importância dos profissionais em discernir infecção de colonização em úlcera venosa, critério fundamental na indicação antibioticoterapia sistêmica.


Subject(s)
Female , Humans , Male , Middle Aged , Anti-Bacterial Agents/pharmacology , Lincosamides/pharmacology , Macrolides/pharmacology , Staphylococcus aureus/drug effects , Streptogramin Group B/pharmacology , Varicose Ulcer/microbiology , Cross-Sectional Studies , Coagulase/metabolism , Drug Resistance, Multiple, Bacterial , Methicillin-Resistant Staphylococcus aureus , Microbial Sensitivity Tests/methods , Phenotype , Prevalence , Primary Health Care , Staphylococcus aureus/classification , Staphylococcus aureus/enzymology
13.
Rev. Inst. Med. Trop. Säo Paulo ; 54(6): 305-306, Nov.-Dec. 2012.
Article in English | LILACS | ID: lil-656263

ABSTRACT

Methicillin-resistant Staphylococcus aureus (MRSA) are now a worldwide problem. Cystic fibrosis (CF) patients are commonly colonized and infected by MRSA. Accurate oxacillin susceptibility testing is mandatory for the adequate management of these patients. We performed a comparison of the accuracy of different tests in CF isolates, including methicillin-susceptible S. aureus and MRSA with different SCCmec types, and using the mecA gene as the gold-standard. The sensitivity and specificity of oxacillin disc, Etest, and oxacillin agar screening plate were 100%. Sensitivity of the cefoxitin disc was 85% and specificity was 100%. For clinically relevant isolates, laboratories may consider the use of a combination of two phenotypic methods.


Staphylococcus aureus resistentes à oxacilina (MRSA) são, atualmente, um problema global. Pacientes com fibrose cística (FC) são frequentemente colonizados e infectados por MRSA. A realização de testes de susceptibilidade acurados é extremamente importante para o manejo da terapia antimicrobiana nesses indivíduos. Nesse estudo, realizamos comparação entre as acurácias de diversos testes de susceptibilidade à oxacilina, em cepas de S. aureus isoladas de pacientes com fibrose cística, tanto sensíveis como resistentes à oxacilina, com diferentes tipos de SCCmec, e utilizando a detecção do gene mecA como método padrão. A sensibilidade e a especificidade do disco de oxacilina, do Etest, e da placa de agar screening com oxacilina foram de 100%. A sensibilidade do disco de cefoxitina foi 85%, com especificidade de 100%. Em cepas clinicamente relevantes, a utilização combinada de mais de um método deveria ser considerada.


Subject(s)
Humans , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Cystic Fibrosis/microbiology , Oxacillin/pharmacology , Staphylococcal Infections/microbiology , Staphylococcus aureus/drug effects , Culture Media/chemistry , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/genetics , Microbial Sensitivity Tests/methods , Phenotype , Reproducibility of Results , Sensitivity and Specificity , Staphylococcus aureus/classification
15.
Indian J Med Microbiol ; 2012 Apr-June; 30(2): 215-217
Article in English | IMSEAR | ID: sea-143949

ABSTRACT

The development of reduced vancomycin susceptibility in Staphylococcus aureus in many cases appears to be associated with characteristic changes. These changes may have pitfall of identifying S. aureus by automated testing methods like Vitek 32. In this study, we retested 24 heterogeneous vancomycin-intermediate Staphylococcus haemolyticus (h-VISH) collected in 2008-2010 at the Department of Clinical Microbiology by conventional biochemical tests and polymerase chain reaction (PCR). The heterogeneous vancomycin-intermediate S. aureus (hVISA) reversion test and electron microscopic examination were also used. Six isolates of 24 h-VISH possessed nuc, coa, and 16S rRNA genes, and could be reversed into S. aureus. It suggested that biochemical and morphological changes in hVISA and vancomycin-intermediate S. aureus (VISA) should be considered, and the detection of S. aureus, especially reduced vancomycin susceptibility isolates, requires more attention and different techniques.


Subject(s)
Bacterial Typing Techniques , Diagnostic Errors , Humans , Microscopy, Electron , Molecular Typing , Polymerase Chain Reaction , Staphylococcal Infections/diagnosis , Staphylococcal Infections/microbiology , Staphylococcus aureus/classification , Staphylococcus aureus/drug effects , Staphylococcus aureus/genetics , Staphylococcus aureus/isolation & purification , Vancomycin Resistance
16.
Article in English | WPRIM | ID: wpr-174789

ABSTRACT

Sensitivity to commercial teat dips (nonoxinol-9 iodine complex and chlorhexidine digluconate) of 56 Staphylococcus (S.) aureus strains isolated from quarter milk samples of various German dairy herds treated with different teat dipping schemes was investigated in this study. The minimum inhibitory concentration was determined using a broth macrodilution method according to the German Veterinary Association guidelines. The main objective of the current study was to induce in vitro resistance induction of S. aureus to chemical disinfectants. Ten different strains were repeatedly passed ten times in growth media with sub-lethal concentrations of disinfectants. Nine strains showed a significant reduction in susceptibility to the nonoxinol-9 iodine complex but only one strain developed resistance to chlorhexidine digluconate. Stability of the acquired resistance was observed in all S. aureus strains adapted to the nonoxinol-9 iodine complex and chlorhexidine digluconate. In contrast, simultaneous resistance to different antibiotics was not observed in any of the ten investigated S. aureus strains. However, the isolates exhibited a high degree of resistance to penicillin G. Based on these results, resistance of S. aureus to chemical disinfectants may be more likely to develop if the chemicals are used at concentrations lower than that required for an optimal biocidal effect.


Subject(s)
Animals , Anti-Bacterial Agents/pharmacology , Cattle , Chlorhexidine/pharmacology , Disinfectants/pharmacology , Dose-Response Relationship, Drug , Drug Resistance, Bacterial , Female , Germany/epidemiology , Iodine/chemistry , Mastitis, Bovine/epidemiology , Nonoxynol/pharmacology , Staphylococcal Infections/epidemiology , Staphylococcus aureus/classification
17.
Article in English | WPRIM | ID: wpr-43986

ABSTRACT

BACKGROUND: Multiple-locus variable-number tandem-repeat fingerprinting (MLVF) is based on multiplex PCR, utilizing variable number tandem repeat. Our goal was to compare the performance of MLVF in distinguishing clinical Staphylococcus aureus isolates with that of pulsed-field gel electrophoresis (PFGE), which has traditionally been the gold standard. METHODS: Sixty-three clinically significant S. aureus isolates were tested using both PFGE and MLVF. Multiplex PCR for MLVF was performed using PCR primers for clfA, clfB, sdrCDE, sspA, and spa. PFGE was performed with genomic DNA fragments generated by SmaI endonuclease digestion. Banding patterns of MLVF or PFGE were analyzed using InfoQuestFP software. RESULTS: The hands-on time of our modified method was about 3 h, on average, for each of 18 isolates. PFGE (80% cutoff) or MLVF (75% cutoff) separated all of the 63 isolates into 13 and 12 types, respectively. Three types generated by PFGE were identical to those generated by MLVF. PFGE and MLVF yielded similar Simpson's diversity indices, indicating similar discriminatory power. The overall concordance between PFGE and MLVF was low, as represented by adjusted Rand indices (0.266-0.278). PFGE predicted MLVF type better than MLVF predicted PFGE type, as reflected by Wallace coefficients (PFGE cutoff 80% vs. MLVF cutoff 75%, 0.389 vs. 0.233). Analysis of the relationship between a pair of isolates showed 91.0% concordance between the PFGE (80% cutoff) and MLVF (75% cutoff). CONCLUSIONS: Our simple, low-cost, modified MLVF protocol can effectively discriminate between S. aureus clinical isolates. MLVF can replace PFGE for the hospital infection control of S. aureus.


Subject(s)
Bacterial Typing Techniques/methods , DNA Fingerprinting , DNA, Bacterial/analysis , Electrophoresis, Gel, Pulsed-Field , Genotype , Humans , Methicillin-Resistant Staphylococcus aureus/classification , Multiplex Polymerase Chain Reaction , Staphylococcal Infections/microbiology , Staphylococcus aureus/classification
18.
Article in English | WPRIM | ID: wpr-47183

ABSTRACT

Staphylococcus (S.) aureus is a common infectious agent of bovine chronic mastitis, a disease that is difficult to eradicate. The abilities of Staphylococci to be internalized and form a biofilm can contribute to host immunological defence evasion that subsequently impairs antimicrobial therapy. The invasive capability of six S. aureus field isolates with different biofilm-forming profiles was compared in vitro using a bovine mammary epithelial cell line. This was further confirmed in primary cell cultures using fluorescent rRNA probes against S. aureus. The results suggest that S. aureus invasion levels are not related to biofilm formation.


Subject(s)
Animals , Biofilms , Cattle , Cell Line , Colony Count, Microbial/veterinary , Epithelial Cells/microbiology , Female , In Situ Hybridization, Fluorescence , Mastitis, Bovine/microbiology , Portugal , Staphylococcal Infections/veterinary , Staphylococcus aureus/classification , Virulence Factors
19.
Article in English | WPRIM | ID: wpr-221270

ABSTRACT

Staphylococcus aureus is a major etiological pathogen of bovine mastitis, which triggers significant economic losses in dairy herds worldwide. In this study, S. aureus strains isolated from the milk of cows suffering from mastitis in Korea were investigated by spa typing and staphylococcal enterotoxin (SE) gene profiling. Forty-four S. aureus strains were isolated from 26 farms in five provinces. All isolates grouped into five clusters and two singletons based on 14 spa types. Cluster 1 and 2 isolates comprised 38.6% and 36.4% of total isolates, respectively, which were distributed in more than four provinces. SE and SE-like toxin genes were detected in 34 (77.3%) isolates and the most frequently detected SE gene profile was seg, sei, selm, seln, and selo genes (16 isolates, 36.3%), which was comparable to one of the genomic islands, Type I nuSabeta. This is a first report of spa types and the prevalence of the recently described SE and SE-like toxin genes among S. aureus isolates from bovine raw milk in Korea. Two predominant spa groups were distributed widely and recently described SE and SE-like toxin genes were detected frequently.


Subject(s)
Animals , Cattle , Cluster Analysis , DNA, Bacterial/chemistry , Enterotoxins/chemistry , Female , Genotype , Mastitis, Bovine/epidemiology , Microsatellite Repeats , Milk/microbiology , Polymerase Chain Reaction/veterinary , Prevalence , Republic of Korea/epidemiology , Staphylococcal Infections/epidemiology , Staphylococcus aureus/classification
20.
Rev. argent. microbiol ; 40(4): 198-203, oct.-dic. 2008. ilus, graf, tab
Article in Spanish | LILACS | ID: lil-634601

ABSTRACT

En febrero de 2006 ocurrió un brote epidémico de gastroenteritis aguda de origen alimentario, en ocasión de un festejo popular en una pequeña localidad de la provincia de Neuquén, Argentina. Aproximadamente 800 personas participaron de un almuerzo en las instalaciones del Gimnasio Municipal, y unas tres horas después de finalizado, cerca de 150 asistentes consultaron al hospital local, afectados por síndrome gastroentérico agudo. Se realizó una investigación epidemiológica caso-control a través de un muestreo representativo no probabilístico. Los resultados epidemiológicos establecieron un brote de ETA a fuente común, con una relación caso-control de 1:1,8. Los principales síntomas fueron cólicos abdominales (88%), vómitos (73,5%) y diarrea (60%). La torta que se sirvió en ese evento fue identificada como el alimento causal (OR 9,79; IC 95%; 2,66-36,00; valor p = 0,0001), sujeto a condiciones higiénico-sanitarias insatisfactorias en los diferentes procesos de elaboración, conservación y manipulación. De una porción de la torta se aisló una cepa de Staphylococcus aureus subespecie aureus, coagulasa positiva, enterotoxigénica, con un recuento de 2,4x10(6) UFC/g, y también se aisló este microorganismo de tres muestras de manos y narinas de personas involucradas en la preparación y el servicio. Las cepas aisladas de un operador y de la torta portaron el gen sea y presentaron el mismo patrón de SmaI-PFGE. Se atribuyó el brote de ETA a la contaminación durante el proceso de preparación de la torta consumida durante ese almuerzo popular, lo que podría estar relacionado con deficiencias en aspectos higiénicos y con la falta de refrigeración y de mantenimiento de la cadena de frío.


In the summer of 2006, an epidemic outbreak of acute gastrointestinal illness related to food consumption occurred in a small town in the province of Neuquén, Argentina. During a popular feast, approximately 800 local residents attended lunch held in the facilities of the Municipal Gymnasium. About three hours later, nearly 150 attendees sought medical assistance at the local hospital due to acute gastroenteritis. A case-control epidemiological investigation was conducted using representative non-probability sampling. The epidemiological investigation showed a common-source foodborne disease outbreak with a case-control ratio of 1:1.8. The main symptoms were abdominal cramps (88%), vomiting (73.5%) and diarrhea (60%). The cake was identified as the source of infection (OR 9.79; IC 95%, 2.66-36.00; p = 0.0001), and unsatisfactory hygienic conditions in food production, conservation and handling steps were identified. Coagulase positive, enterotoxigenic Staphylococcus aureus, subspecies aureus was detected in a piece of cake, with a count of 2.4x10(6) CFU/g, and in samples from the hands and nostrils of three people involved in food preparation and service. The strains isolated from both the cake and one of the food handlers carried the sea gene, and presented the same SmaI-PFGE pattern. The foodborne disease outbreak was considered to be due to contamination in the preparation process of the cake consumed at the feast, which was related to inadequate hygienic conditions, lack of refrigeration and cold chain disruption.


Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Infant , Male , Middle Aged , Disease Outbreaks , Food Contamination , Gastroenteritis/epidemiology , Staphylococcal Food Poisoning/epidemiology , Argentina/epidemiology , Bacterial Typing Techniques , Case-Control Studies , Carrier State/metabolism , Food Handling , Gastroenteritis/etiology , Gastroenteritis/microbiology , Refrigeration , Staphylococcal Food Poisoning/microbiology , Staphylococcus aureus/classification , Staphylococcus aureus/genetics , Staphylococcus aureus/isolation & purification
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