Ocorrência e determinação do perfil de sensibilidade às drogas das cepas de Staphylococcus aureus isoladas de fossas nasais e orofaringe dos servidores hospitalares

Staphylococcus aureus is a very important hospital and community pathogen. This species is related to supurative disease, systemic and widespread metastatic lesions. The ability of S. aureus to develop resistance to antibiotics, more recently to methicillin, associates this bacteria with epidemic outbreaks of severe nosocomial infection. The source of staphylococcal infection is a patient with a staphylococcal lesion or a career member of the hospital staff. We aimed to detect the frequency of S. aureus isolated from anterior nares and oral cavities among the hospital staff in Bauru - SP, and to determine the antibiotic susceptibility of the isolates. Within 213 of the staff members analyzed. S. aureus was found in 94 (44.13%) of careers, with 47 (50%) of nasal carriers, 23 (24.4%) of oral carriers and 24 (25.5%) of both carriers. The biochemical characteristics analyzed for the species identification were similar to S. aureus ATCC 29213. All strains identified as S. aureus showed varied sensibility to the antimicrobial agents tested. No vancomicin resistant strains and only 8 (8.5%) strains with oxacilin resistance were found.

Atividade anti Staphylococcus aureus de extratos de própolis (EP) de Apis mellifera preparados com diferentes concentrações de etanol

Propolis has been used in folk medicine and possesses a broad spectrum of biological activities, specially antibacterial activity. Studies have demonstrated that the composition of propolis extract may have influence in such activity. The goal of this study was to investigate the antibacterial activity of eleven propolis extracts (PE) against sixty one Staphylococcus aureus strains, isolated from newborn clinical specimens. The PE from Apis mellifera were prepared by using pure water and mixtures of water with ethanol at different concentrations (from 0 to 100%), 25g of propolis in 100 mL of solvent, and three days of maceration followed by filtration. Determination of Minimal Inhibitory Concentration (MIC) by agar dilution method was performed and serial concentrations from each PE were achieved (%v/v) in plates containing Mueller Hinton agar. It was possible to verify that the anti S. aureus activity was directly proportional to ethanol concentration and no significant differences were observed among PE with ethanol concentration from 70 to 100%. The MIC 90% values ranged from 0.4 to 0.6% (v/v) and the 70% ethanolic extract were the most efficient to inhibit bacterial growth (MIC 90%=0.42%, v/v). In conclusion, our results suggest that the EP composition and, consequently, the concentration of ethanol used as solvent may influence the antibacterial activity of propolis from A. mellifera.

Pheno and genotyping of Staphylococcus aureus, isolated from bovine milk samples from São Paulo State, Brazil

In the present study, 87 Staphylococcus aureus isolates obtained from milk samples of 87 cows with mastitis in 6 different municipal districts of 2 regions of São Paulo State, Brazil, were compared pheno and genotypically. Pulsed-field gel electrophoresis (PFGE) analysis of the strains was performed, and PCR was carried out to detect genes for a number of staphylococcal cell surface proteins, exoproteins, and 3 classes of agr genes. Nine distinct S. aureus lineages (LA-LI) were identified by PFGE. The lineages LA and LE, which accounted together for 63 strains (72.2%), were prevalent and had been collected from all of the 6 municipal districts, indicating a broad geographic distribution of these lineages; LB, LC, LD, LF, LG, LH, and LI, however, were isolated sporadically and accounted for 24 strains (27.8%). Some characteristics, like penicillin resistance and the presence of cap8 and agr class II genes, were associated with the prevalent lineages (LA and LE), and penicillin susceptibility and the presence of cna and cap5 genes were associated with sporadic lineages. According to the present results, some S. aureus lineages possess a combination of genes that confer the propensity to cause and disseminate infection, and only a limited number of clones are responsible for the cases of bovine mastitis on the various farms. © 2004 NRC Canada.

Aderência e formação do biofilme por Staphylococcus aureus ao polietilenotereftalato (PET) usado como embalagem para acondionamento de medicamentos e cosméticos

The polyethylene terephthalate (PET) is used for package drugs and cosmetics. The aim of this research was examine by scanning electron microscopy (SEM) a Staphylococcus aureus attachment and biofilm formation on a polyethylene terephthalate (PET) surface and hydrophobicity of S. aureus by adherence to hydrocarbons. A suspension of S. aureus was prepared in Mueller-Hinton broth and, coupons of polyethylene terephthalate were incubated for 30 minutes, two, 24 and 48 hours, 15 and 30 days. Afteron the coupons were removed and prepared for scanning electron microscope analysis. The attachment and biofilm formation was observed on the surfaces of PET. The SEM revealed adhesion and biofilm formation on PET surfaces. The hydrophobicity test classified S. aureus as hydrophobic.

Propolis: anti-Staphylococcus aureus activity and synergism with antimicrobial drugs

Propolis is a natural resinous substance collected by bees from tree exudates and secretions. Its antimicrobial activity has been investigated and inhibitory action on Staphylococcus aureus growth was evaluated The in vitro synergism between ethanolic extract of propolis (EEP) and antimicrobial drugs by two susceptibility tests (Kirby and Bauer and E-Test) on 25 S. aureus strains was evaluated Petri dishes with sub-inhibitory concentrations of EEP were incubated with 13 drugs using Kirby and Bauer method and synergism between EEP and five drugs [choramphenicol (CLO), gentamicin (GEN), netilmicin (NET), tetracycline (TET), and vancomycin (VAN)] was observed. Nine drugs were assayed by the E-test method and five of them exhibited a synergism [CLO, GEN, NET, TET, and clindamycin (CLI)]. The results demonstrated the synergism between EEP and antimicrobial drugs, especially those agents that interfere on bacterial protein synthesis.

Evaluation of the antimicrobial susceptibility of methicillin-resistant Staphylococcus aureus isolated from human infections

Multiresistant Staphylococcus aureus constitutes an important public health problem, especially in view of its possible spread in nosocomial environments. In the present work, we analyzed the susceptibility profile of 80 S. aureus stains from human infections resistant to at least 10 drugs. For this study, the techniques used were the disk method and minimum inhibitory concentration (MIC) of the following drugs: cefuroxime, ciprofloxacin, clindamycin, erythromycin, gentamycin, imipenem, oxacillin, rifampicin, tetracycline and vancomycin, according the criteria of the National Committee for Clinical Laboratory Standards (NCCLS). Methicillin was included in the antibiogram as a marker, which is usually used in drugs selection for the treatment of staphylococcal infections. Results indicated that the most effective drug was vancomycin. For the other 10 drugs, the percentage of resistant strains ranged from 85% to 93.75%. In relation to the MICs, it was observed that vancomycin (MIC 90% = 0.615ug/ml) was the most effective drug; followed by rifampicin (MIC 90% = 2.6ug/ml) and ciprofloxacin (MIC 90% = 26.6ug/ml). The drugs that showed the least effective activity were cefuroxime, clindamycin, erythromycin, gentamycin, and oxacillin. On the other hand, observation of β-lactamase production revealed that most of the methicillin-resistant strains produced β-lactamase (83.7%), potentiating the risks of nosocomial infections. In general, vancomycin still continues to be one of the most effective drugs for staphylococcal infections therapy.

Risk of peritonitis during peritoneal dialysis in carriers of Staphylococcus aureus and coagulase-negative staphylococci

The presence of Staphylococcus aureus in the nasal cavities and pericatheter skin of peritoneal dialysis patients put them at high risk of developing peritonitis. However, it is not clear whether the presence of coagulase-negative staphylococci (CNS) in the nasal passages and skin of patients is related to subsequent occurrence of peritoneal infection. The aim of the present study was to verify the relationship between endogenous sources of S. aureus and CNS and occurrence of peritonitis in patients undergoing peritoneal dialysis. Thirty-two patients on peritoneal hemodialysis were observed for 18 months. Staphylococcus species present in their nasal passage, pericatheter skin and peritoneal effluent were identified and compared based on drug susceptibility tests and dendrograms, which were drawn to better visualize the similarity among strains from extraperitoneal sites as well as their involvement in the causes of infection. Out of 288 Staphylococcus strains isolated, 155 (53.8%) were detected in the nasal cavity, 122 (42.4%) on the skin, and 11 (3.8%) in the peritoneal effluent of patients who developed peritonitis during the study. The most frequent Staphylococcus species were CNS (78.1%), compared with S. aureus (21.9%). Among CNS, S. epidermidis was predominant (64.4%), followed by S. warneri (15.1%), S. haemolyticus (10.7%), and other species (9.8%). Seven (64%) out of 11 cases of peritonitis analyzed presented similar strains. The same strain was isolated from different sites in two (66%) out of three S. aureus infection cases. In the six cases of S. epidermidis peritonitis, the species that caused infection was also found in the normal flora. From these, two cases (33%) presented highly similar strains and in three cases (50%), it was difficult to group strains as to similarity. Patients colonized with multidrug-resistant S. epidermidis strains were more predisposed to infection. Results demonstrated that an endogenous source of S. epidermidis could cause peritonitis in peritoneal dialysis patients, similarly to what has been observed with S. aureus.

Anatomia do músculo abdutor crural caudal do gato doméstico (Felis catus domesticus, Linnaeus 1758)

The frequency and the morphology of the abductor cruris caudalis muscle were studied in 58 adult unknown breed cats. The sample included 36 females and 22 males. After the dissection, were found a frequency for the presence of the same muscle of 98.3%. The abductor cruris caudalis muscle appears symmetrically in 35 females (97.2%) and 22 males (100.0%). When present, it were originated on the transversal process of the 1st vertebrae caudales, in the glutea region, below the gluteus superficialis muscle, throughing almost all the thigh under biceps femoris muscle, where it make a parallel relation with the sciatic nerve, receiving nervous ramifications of the same in 78.3% of the cases. The medium found for the width and thickness of his muscle venter was respectively of 2.03 mm and 0.35 mm. Its insertion occurs on the biceps femoris muscles approximate to the fascia cruris, where both finish fusing theirselves.

Research on staphylococcus spp in biofilm formations in water pipes and sinsibility to antibiotics

Water from dental equipment presents risks for surgeon-dentists as well as for patients because it might work as a means of dissemination/ transmission of microoganisms. The objective of this study was to verify the quality of the water used in dental equipment by means of microbiological analysis, accomplishing the count of Staphylococcus spp.There have been collected, 160 samples of water from reserviors, taps used for hand washing, air-water syringes, and high-speed handpieces, in 40 dental offices in the city of Barretos, São Paulo. The rules concerning bacteriotogicaI analysis in cfu/mL from Standard Methods for the Examination of Water and Wastewater have been followed. The analysis of the results has made it possible to verify that out of the total of samples, 28% did not meet the standards of potability established by the American Dental Association: Regarding the origin of analyzed S. aureus., the most contaminated sites were high-speed handpicces in private offices (761%) and in, ental care plan offices (71%), followed by air-water syringe in dental care plan offices (64%). For S. epidermitis samples, the most contaminated sites were high-speed handpieces in SUS (Brazilian Government Health System) dental offices (22%) and in dental care plan offices (14%) The most contaminated sites were dental offices that saw Patients under dental care plans, Concerning tested antibiotics, the ones that presented better results as to sensibility to strain S. epidermidis were vancomycin and ciprofloxacin (100%) and, as to sensibility to strain S. aureus, it was ciprofloxacin (97%).

Detection of enterotoxin and toxic shock syndrome toxin 1 genes in Staphylococcus, with emphasis on coagulase-negative staphylococci

The detection of staphylococcal enterotoxins is decisive for the confirmation of an outbreak and for the determination of the enterotoxigenicity of strains. Since the recognition of their antigenicity, a large number of serological methods for the detection of enterotoxins in food and culture media have been proposed. Since immunological methods require detectable amounts of toxin, molecular biology techniques represent important tools in the microbiology laboratory. In the present study, polymerase chain reaction (PCR) was used to identify genes responsible for the production of enterotoxins and toxic shock syndrome toxin 1 (TSST-1) in S. aureus and coagulase-negative staphylococci (CNS) isolated from patients and the results were compared with those obtained by the reverse passive latex agglutination (RPLA) assay. PCR detection of toxin genes revealed a higher percentage of toxigenic S. aureus strains (46.7%) than the RPLA method (38.3%). Analysis of the toxigenic profile of CNS strains showed that 26.7% of the isolates produced some type of toxin, and one or more toxin-specific genes were detected in 40% of the isolates. These results suggests the need for further studies in order to better characterize the pathogenic potential of CNS and indicate that attention should be paid to the toxigenic capacity of this group of microorganisms.

Mastite subclínica causada por Staphylococcus aureus: Custo-benefício da antibioticoterapia de vacas em lactação

Economic evaluation of the treatment bovine subclinical mastitis caused by S. aureus was evaluated. Two hundred and seventy udder quarters with or without subclinical mastitis were distributed into four groups, in conformity to lactational stage and treatments. Group 1 included animals treated between 10 and 60 days of lactation; group 2 included animals treated from 61 days of lactation to two months before drying; group 3 included animals no treated between 10 and 60 days of lactation; group 4 included animals no treated from 61 days of lactation to two months before drying. Treatment with gentamicin (150mg) was accomplished by intramammary doses, once a day, after performing sensitivity tests. The mammary quarters were re-evaluated after 30 days. The costs with the treatment were calculated considering a S. aureus prevalence of 5% as well as expenses with antibiotic, milk disposal, tests of drug sensitivity and workload. There was loss of income of 2% and 14% in the groups 1 and 2, respectively, when compared with the values before the treatment. In such case, the treatment of bovine subclinical mastitis caused by S. aureus in the lactation was economically unviable.

Subclinical mastitis caused by staphylococcus aureus: Analysis cost benefit of antibiotic therapy in lactating cows

The cost benefit analysis of treatment of bovine subclinical mastitis caused by S. aureus was evaluated. Two hundred and seventy udder quarters with subclinical mastitis and healthy were selected in four groups, in conformity to lactational stage and with the treatment or not. Group 1 included treated animals 10 to 60 days in milk; group 2 included treated animals 61 days in milk until two months before the end of lactation; group 3 included animals not treated 10 to 60 days in milk; group 4 included animals not treated from 61 days in milk until two months before the end of lactation. Treatment with gentamicin (150 mg) was accomplished by intramammary doses, once a day, after sensitivity tests. The mammary quarters were evaluated after 30 days again. The costs with the treatment were calculated considering a S. aureus prevalence of 5%, expenses with antibiotic, loss in milk, tests of sensitivity and workload. There was loss of income of 2% and 14% in the groups 1 and 2, respectively, when compared with the incomes before treatment. In such case, the treatment of bovine subclinical mastitis by S. aureus in the lactation was economically not practicable.

Determinação da resistência de Staphylococcus aureus: Um desafio?

The aim of this study was to identify the resistance profile of Staphylococcus aureus strains, in relation to induced clindamycin resistance, and to detect oxacillin resistance by the routine phenotypic methods. The strains were isolated from nasal or lingual swabs taken from healthy adult carriers with no medical history of hospitalization or antibiotic treatment. Eighteen strains were distinguished by the different patterns generated by pulsed field gel electrophoresis (PFGE). Four (22.2%) of these showed sensitivity to clindamycin by the conventional antibacterial susceptibility test, but demonstrated inducible resistance to it by the D-test. One strain (5.6%) was characterized as borderline oxacillin-resistant S. aureus (BORSA), and another (5.6%) as CA MRSA (community-associated methicillin-resistant Staphylococcus aureus). Both of these strains were shown to be cefoxitin susceptible by the disk diffusion test. The polymerase chain reaction (PCR) failed to detect the mecA gene in this last strain and it was thus classified as BORSA. These results show the importance of incorporating the D-test into the routine lab tests for S. aureus inducible clindamycin resistance and also of including the cefoxitin resistance test among the phenotypic methods for MRSA characterization.

Multiplex PCR use for Staphylococcus aureus identification and oxacillin and mupirocin resistance evaluation

Oxacillin-resistant Staphylococcus aureus represents a serious problem in hospitals worldwide, increasing infected patients' mortality and morbidity and raising treatment costs and internment time. In this study, the results of using the Multiplex PCR technique to amplify fragments of the genes femA (specific-species), mecA (oxacillin resistance) and ileS-2 (mupirocin resistance) were compared with those of tests conventionally used to identify S. aureus isolates and ascertain their resistance to drugs. Fifty S. aureus strains were isolated from patients receiving treatment at UNOESTE University Hospital in Presidente Prudente, SP, Brazil. The 686 bp fragment corresponding to the gene femA was amplified and detected in all the isolates. On the other hand, the 310 bp fragment corresponding to the mecA gene was amplified in 29 (58%) of the isolates. All of the isolates showed sensitivity to mupirocin in the agar diffusion test, which was corroborated by the lack of any amplicon of the 456 bp fragment corresponding to the ileS-2 gene, in the PCR bands. The conventional tests to identify S. aureus and detect resistance to oxacillin and mupirocin showed 100% agreement with the PCR Multiplex results. The use of techniques for rapid and accurate identification of bacteria and assessment of their resistance may be valuable in the control of infection by resistant strains, allowing the rapid isolation and treatment of an infected patient. However, the results demonstrate that traditional phenotypic tests are also reliable, though they take more time.

Quercetin reduces Staphylococcus aureus interaction with neutrophils

Flavonoids, including quercetin, have been reported to modulate the ability of Staphylococcus aureus to adhere to host tissue without exhibiting direct antibacterial activity. In the present study, we evaluated the interaction of S. aureus pretreated with 40 μg/mL of quercetin with neutrophils to assay oxidative burst stimulation, using luminol-amplified chemiluminescence. S. aureus pre-incubated with subinhibitory concentration of quercetin induced significantly less light emission by neutrophils than did untreated bacteria. The results of the present study demonstrate that quercetin decreases S. aureus uptake by neutrophils.