Phenotypic and molecular methods were used to characterize the antibiotic resistance of 64 clinical isolates of Staphylococcus haemolyticus. By PCR of the mecA gene, 87% were found to be methicillin resistant. Approximately 55% harbored staphylococcal cassette chromosome mec element (SCCmec) type V, and only one SCCmec type IV. Many isolates (75%) displayed multiresistance, and pulsotype analysis showed a high diversity.A mong coagulase-negative staphylococci (CoNS), Staphylococcus haemolyticus is the second most frequently isolated from human blood cultures (18) and has the highest level of antimicrobial resistance (3,8). Methicillin resistance is conferred by the mecA gene, carried on the staphylococcal cassette chromosome mec element (SCCmec) (12). Eight types (I to VIII) of SCCmec have been assigned for Staphylococcus aureus (11), and SCCmec type V has already been found in CoNS, particularly in S. haemolyticus (13). The increase in the frequency of methicillinresistant S. haemolyticus as the causal agent of hospital infections and the possibility of emergence of resistance to other antibiotics demand trustworthy characterization of the isolates and an investigation of clonal spreading within hospitals.In the studies reported here, 64 clinical strains were isolated from patients at Hospital Naval Marcílio Dias, Rio de Janeiro, Brazil, between 2006 and 2008. The strains were isolated from the following clinical infections or sources in 31 males and 33 females: bacteremia (n ϭ 45), skin (n ϭ 2), urine (n ϭ 13), and unknown source (n ϭ 4). The isolates were identified at the hospital laboratory as S. haemolyticus by using the MicroScan WalkAway PC21 panel, and their identification was confirmed by specific PCR (17).The resistance profiles of the strains for the main antibiotics used in Brazil were determined by disc diffusion tests according to CLSI guidelines (5). However, the mupirocin susceptibility testing was not preconized by CLSI, so the results for this antibiotic were interpreted as previously described (7, 9). The methicillin resistance was also evaluated by other phenotypic methods, such as the MIC for oxacillin (5), the MicroScan, and PCR of the mecA gene (6). The SCCmec type was determined in a multiplex PCR as previously described (14), except that the pair of primers mecI P2 and mecI P3 used as the internal control were replaced by MRS1 and MRS2 (6), which amplify a 154-bp fragment of the mecA gene.Analysis of genetic relatedness and characterization of isolates using pulsed-field gel electrophoresis (PFGE) of genomic DNA digested with SmaI were carried out as previously described (20). Banding patterns were determined by visual inspection and by using Bionumerics software, version 6.0 (Applied Maths) using the Dice index and the unweighted-pair group method with arithmetic average. Similar PFGE genotypes were defined using a coefficient of similarity of up to 80%, and the subtypes were those with less than five-band variants, as recommended by van Belkun et al. (19).As shown in Table 1, there wa...

The genetic basis of bacteriocin (Bac) production by six strains of Stuphylococcus aureus was examined. Gene transfer experiments (in which the plasmids were tagged with the erythromycin resistance transposon Tn551) and plasmid-elimination experiments by growth at 43 "C associated bacteriocin production with a particular plasmid in each strain. The Bac plasmids could be separated into two distinct groups: the first comprised plasmids larger than 40 kb, which did not specify immunity to bacteriocins; the second comprised small plasmids (8.0-10-4 kb) which also specified immunity to bacteriocins. The sequence relations among the small plasmids (pRJ6, pRJ9, pRJlO and pRJ11) were investigated by comparing restriction enzyme digest patterns and by hybridization. Plasmids pRJlO and pRJ11 were indistinguishable and very closely related to plasmid pRJ9. Plasmid pRJ6, although different from the others, shared regions of sequence homology with them. No homology was found between plasmids pRJ6 or pRJ9 and the large Bac plasmids.

Bacteriocins are bacterial antimicrobial peptides with bactericidal activity against other bacteria. Staphylococcins are bacteriocins produced by staphylococci, which are Gram-positive bacteria with medical and veterinary importance. Most bacteriocins produced by staphylococci are either lantibiotics (e.g., Pep5, epidermin, epilancin K7, epicidin 280, staphylococcin C55/BacR1, and nukacin ISK-1) or class II bacteriocins (e.g., aureocins A70 and 53). Only one staphylococcin belonging to class III, lysostaphin, has been described so far. Production of staphylococcins is a self-protection mechanism that helps staphylococci to survive in their natural habitats. However, since these substances generally have a broad spectrum of activity, inhibiting several human and animal pathogens, they have potential biotechnological applications either as food preservatives or therapeutic agents. Due to the increasing consumer awareness of the risks derived not only from food-borne pathogens, but also from the artificial chemical preservatives used to control them, the interest in the discovery of natural food preservatives has increased considerably. The emergence and dissemination of antibiotic resistance among human and animal pathogens and their association with the use of antibiotics constitute a serious problem worldwide requiring effective measures for controlling their spread. Staphylococcins may be used, solely or in combination with other chemical agents, to avoid food contamination or spoilage and to prevent or treat bacterial infectious diseases. The use of combinations of antimicrobials is common in the clinical setting and expands the spectrum of organisms that can be targeted, prevents the emergence of resistant organisms, decreases toxicity by allowing lower doses of both agents and can result in synergistic inhibition.

Aims:  To investigate the activity of seven staphylococcins, bacteriocins produced by staphylococci, against multiresistant Staphylococcus aureus and coagulase‐negative staphylococci (CNS) involved in human infections. Methods and Results:  Four bacteriocins produced by Staph. epidermidis (Pep5, epidermin, epilancin K7 and epicidin 280) and three produced by Staph. aureus (aureocins A70, A53 and 215FN) were tested. Sixteen Staph. aureus strains, including a representative strain of the endemic Brazilian methicillin‐resistant clone (MRSA), and 57 CNS strains were used as indicators. Among the staphylococcins used, Pep5 was able to inhibit 77·2% of the CNS strains and 87·5% of the Staph. aureus strains tested, including the Brazilian MRSA endemic clone, responsible for a large number of hospital‐acquired infections in Brazil. On the other hand, aureocin A53 and epidermin presented a high antagonistic activity only against the Staph. aureus strains, being able to inhibit, respectively, 87·5% and 81·3% of them, including also the Brazilian MRSA endemic clone. The remaining bacteriocins inhibited only a low percentage of the nosocomial staphylococcal strains tested. Conclusions:  Aureocin A53 and epidermin have potential applications against MRSA, whereas Pep5 seems to be an attractive agent against both MRSA and CNS, including mupirocin‐resistant strains and the Brazilian endemic clone of MRSA, which is also found disseminated in other countries. Significance and Impact of the Study:  Bacteriocins may represent alternative agents to control important nosocomial pathogens.

Aims: The purpose of this study was to determine the susceptibility of Campylobacter jejuni and Campylobacter coli isolates to antimicrobial agents and to investigate the presence of plasmid DNA. Methods and Results: A total of 15 clinical isolates from children faeces, and 29 animal isolates of Campylobacter jejuni (n 22) and Campylobacter coli (n 22) were tested for susceptibility to 9 antimicrobial agents using a disc diffusion method, and screened for the presence of plasmid DNA by agarose gel electrophoresis. Of the 44 isolates, 56á8% were resistant to sulphonamide, 25% to nor¯oxacin, 18á2% to erythromicin, cipro¯oxacin and ampicillin, and 13á6% to tetracycline. All isolates were susceptible to gentamicin, chloramphenicol and cefotaxime. Plasmids were detected in one Camp. jejuni (4á54%) strain isolated from sheep and in six (27á27%) Camp. coli strains isolated from rhesus monkey(3), swine(2), and poultry(1) with sizes ranging from 3á4 to 50 kb. Conclusions: The majority of the human isolates were susceptible to antibiotics commonly used for the treatment of campylobacteriosis. Signi®cance and Impact of the Study: The origin and spread of Campylobacter resistance to antibiotics are discussed, with particular respect to the current situation in Brazil.

In this work, mupirocin resistance was correlated with the presence of plasmids in methicillin-resistant Staphylococcus aureus (MRSA) strains isolated in the Rio de Janeiro Federal University Hospital in Brazil, where topical mupirocin has been used extensively since 1990. Of 19 strains studied, those exhibiting high-level resistance carried a large and relaxable plasmid of about 35 kb. Mupirocin-sensitive derivatives, obtained by growth at 42 C of a strain exhibiting high-level resistance, were devoid of the large plasmid, which was designated pMG1. Mupirocin resistance was transferred to strain RN8411 during overnight filter-matings at low frequencies (7.0 x 10(-9)/donor). The pMG1 plasmid was shown to be responsible for high-level mupirocin resistance in our isolates and to be incompatible with pGO1. Hybridization experiments suggested that mupirocin resistance in pMG1 is due to the presence of the ileS-2 gene. The pMG1 plasmid was successfully and bidirectionally transferred from Staphylococcus aureus to Staphylococcus epidermidis, suggesting that the latter may be a reservoir of this resistance plasmid. No transfer was detected to Staphylococcus haemolyticus. The development of self-transferable high-level mupirocin resistance should be considered when using mupirocin to control the spread of MRSA in hospitals.

The action of lysostaphin, either alone or in combination with nisin, against established staphylococcal biofilm may represent an alternative to bovine mastitis control. However, the duration of the treatment should be considered for its application so that the best effectiveness can be achieved.

The inhibitory activity of five bacteriocin (Bac)-producer strains of Staphylococcus aureus was tested against bacteria pathogenic for cattle. Sixty-five epidemiologically unrelated strains of Staph. aureus involved in bovine mastitis were used as indicators in an agar diffusion test. Bacteriocins produced by four strains could inhibit only a limited number of test organisms. However, all 65 indicator strains proved to be susceptible to the combined action of both bacteriocins encoded by pRJ9, a Bac plasmid found in strain A53. Therefore, the bacteriocins produced by this strain may represent new antimicrobial peptides with potential applications in the prevention and treatment of bovine mastitis.