J Hosp Infect, 2003 Nov, 55(3), 184 - 9 Staphylococcal species in the oral cavity from patients in a regional burns unit; Smith AJ et al.; The aim of this study was to perform a quantitative and qualitative analysis of oral carriage of staphylococci in a range of oral specimens from patients admitted to a regional burns unit . The study recruited 28 patients and reasons for admittance were: burns (46%), skin grafting (39%), lacerations (7%), scalding (4%) and necrotizing fasciitis (4%) . No patient had smoke inhalation injuries or trauma to the oro-pharynx . There were five patients from whom methicillin-sensitive S . aureus (MRSA) could be detected in oral specimens . For three patients only the wound and oral specimens were positive for MRSA . In one patient only the oral specimens were positive for MRSA . There were five patients from whom methicillin-sensitive S . aureus (MRSA) could be detected in the oral specimens . In one patient only the oral specimens were positive for MSSA . Staphylococci could be recovered from the dental plaque, denture and toothbrush specimens with a mean count of 1.1 x 10(4)cfu/mL (range 20-5.3 x 10(4)), 5.4 x 10(3) (range 40-2.1 x 10(4)) and 264 cfu/mL (range 20-500), respectively . Both MSSA and MRSA could be recovered from these specimen types . In one patient only the toothbrush was positive for MRSA and all other oral specimens were negative . This study suggests that staphylococci are not infrequent colonizers of the oral cavity, and that this site may serve as a potential reservoir for transmission to other body sites. Cell, 2003 Oct 17, 115(2), 217 - 28 A "dock, lock, and latch" structural model for a staphylococcal adhesin binding to fibrinogen; Ponnuraj K et al.; Gram-positive pathogens such as staphylococci contain multiple cell wall-anchored proteins that serve as an interface between the microbe and its environment . Some of these proteins act as adhesins and mediate bacterial attachment to host tissues . SdrG is a cell wall-anchored adhesin from Staphylococcus epidermidis that binds to the Bbeta chain of human fibrinogen (Fg) and is necessary and sufficient for bacterial attachment to Fg-coated biomaterials . Here, we present the crystal structures of the ligand binding region of SdrG as an apoprotein and in complex with a synthetic peptide analogous to its binding site in Fg . Analysis of the crystal structures, along with mutational studies of both the protein and of the peptide, reveals that SdrG binds to its ligand with a dynamic "dock, lock, and latch" mechanism . We propose that this mechanism represents a general mode of ligand binding for structurally related cell wall-anchored proteins of gram-positive bacteria. Vet Res, 2003 Sep-Oct, 34(5), 689 - 716 Mastitis of dairy small ruminants; Bergonier D et al.; Staphylococci are the main aetiological agents of small ruminants intramammary infections (IMI), the more frequent isolates being S . aureus in clinical cases and coagulase negative species in subclinical IMI . The clinical IMI, whose annual incidence is usually lower than 5%, mainly occur at the beginning of machine milking and during the first third of lactation . These features constitute small ruminant peculiarities compared to dairy cattle . Small ruminant mastitis is generally a chronic and contagious infection: the primary sources are mammary and cutaneous carriages, and spreading mainly occurs during milking . Somatic cell counts (SCC) represent a valuable tool for prevalence assessment and screening, but predictive values are better in ewes than in goats . Prevention is most often based on milking machine management, sanitation and annual control, and milking technique optimisation . Elimination mainly relies on culling animals exhibiting clinical, chronic and recurrent IMI, and on drying-off intramammary antibiotherapy; this treatment allows a good efficacy and may be used selectively by targeting infected udders only . Heritability values for lactation mean SCC scores are between 0.11 and 0.15 . Effective inclusion of ewe's mastitis resistance in the breeding goal has recently been implemented in France following experimental and large scale estimations of genetic parameters for SCC scores. J Korean Med Sci, 2003 Oct, 18(5), 631 - 6 Multiplex PCR for the detection of genes encoding aminoglycoside modifying enzymes and methicillin resistance among Staphylococcus species; Choi SM et al.; We developed multiplex polymerase chain reaction (PCR) to detect aac(6 ')/aph(2 "), aph(3 ')-IIIa, and ant(4 ')-Ia, the genes encoding the most clinically relevant amino-glycoside modifying enzymes (AME), and simultaneously, the methicillin resistant gene, mecA, in Staphylococcus species . Clinical isolates of 45 S . aureus and 47 coagulase negative staphylococci (CNS) from tertiary university hospitals were tested by conventional susceptibility testing, using the agar dilution method and by multiplex PCR . Of a total of 92 isolates, 61 isolates were found to be methicillin-resistant . Of these, 54 isolates (89%) were found to be harboring mecA . Seventy-five percent of the 92 isolates demonstrated resistance to at least one of the aminoglycosides tested . Moreover, resistance to aminoglycosides was closely associated with methicillin-resistance (p<0.05) . The most prevalent AME gene was aac(6 ')/aph(2 ") which was found in 65% of the isolates, and ant(4 ')-Ia and aph(3 ')-IIIa were present in 41% and 9% of the isolates, respectively . The concordance between methicillin-resistance and the presence of mecA gene was 98% in S . aureus and 81% in CNS . The concordance between gentamicin resistance and the presence of aac(6 ')/aph(2 ") gene was 100% in S . aureus and 85% in CNS . The multiplex PCR method that we developed appears to be both a more rapid and reliable than conventional method. J Clin Microbiol, 2003 Oct, 41(10), 4740 - 4 Practical disk diffusion method for detection of inducible clindamycin resistance in Staphylococcus aureus and coagulase-negative staphylococci; Fiebelkorn KR et al.; Resistance to macrolides in staphylococci may be due to active efflux (encoded by msrA) or ribosomal target modification (macrolide-lincosamide-streptogramin B {MLSB} resistance; usually encoded by ermA or ermC) . MLSB resistance is either constitutive or inducible following exposure to a macrolide . Induction tests utilize closely approximated erythromycin and clindamycin disks; the flattening of the clindamycin zone adjacent to the erythromycin disk indicates inducible MLSB resistance . The present study reassessed the reliability of placing erythromycin and clindamycin disks in adjacent positions (26 to 28 mm apart) in a standard disk dispenser, compared to distances of 15 or 20 mm . A group of 130 clinical isolates of Staphylococcus aureus and 100 isolates of erythromycin-resistant coagulase-negative staphylococci (CNS) were examined by disk approximation; all CNS isolates and a subset of S . aureus isolates were examined by PCR for ermA, ermC, and msrA . Of 114 erythromycin-resistant S . aureus isolates, 39 demonstrated constitutive resistance to clindamycin, while 33 showed inducible resistance by disk approximation at all three distances . Only one isolate failed to clearly demonstrate induction at 26 mm . Of 82 erythromycin-resistant CNS isolates that contained ermA or ermC, 57 demonstrated constitutive clindamycin resistance, and 25 demonstrated inducible resistance, at 20 and 26 mm . None of the 42 S . aureus isolates or 18 CNS isolates containing only msrA and none of the erythromycin-susceptible isolates yielded positive disk approximation tests . Simple placement of erythromycin and clindamycin disks at a distance achieved with a standard disk dispenser allowed detection of 97% of S . aureus strains and 100% of CNS strains with inducible MLSB resistance in this study. J Clin Microbiol, 2003 Oct, 41(10), 4660 - 5 Decreased vancomycin susceptibility of coagulase-negative staphylococci in a neonatal intensive care unit: evidence of spread of Staphylococcus warneri; Center KJ et al.; Coagulase-negative staphylococci (CoNS) are important pathogens in premature neonates; decreasing glycopeptide susceptibility has been observed among these isolates . The epidemiology of colonization with CoNS, the organisms' vancomycin susceptibilities, and genetic relatedness were studied over 6 months in a tertiary-care neonatal unit . A total of 321 isolates of CoNS were isolated . Seventy-five percent of the infants were colonized at admission, and virtually all were colonized thereafter . Common species were Staphylococcus epidermidis (69%), S . warneri (12%), S . haemolyticus (9.7%), and S . hominis (5.6%) . A total of 3.9% of CoNS isolates had decreased vancomycin susceptibility (DVS) (MICs > 2.0 microg/ml); isolate recovery was associated with a stay in a neonatal intensive care unit for >28 days (P = 0.039), vancomycin exposure (P = 0.021), and S . warneri colonization (P < 0.0001) . Nine of 12 (75%) CoNS with DVS were S . warneri, had enhanceable high-level resistance in vitro, were indistinguishable or closely related by pulsed-field gel electrophoresis, and were different from 29 vancomycin-susceptible S . warneri isolates . Epidemiological analysis suggested unsuspected nosocomial spread . Species determination in certain settings may aid in the understanding of emerging nosocomial problems. Transplant Proc, 2003 Sep, 35(6), 2361 - 2 The effect of cyclosporine and tacrolimus on indigenous bacterial flora in human skin grafts; Moscicka M et al.; Allogeneic skin transplants require intensive immunosuppressive therapy . Treatment protocols used for parenchymal organ grafts are not satisfactory to prevent skin graft rejection . Another factor responsible for the destruction of allogeneic skin transplants is bacterial inflammation . Temporary ischemia and the allogeneic reaction in transplanted skin cause increased permeability of the epidermis and the dermal capillaries, making skin grafts vulnerable to bacterial penetration . Moreover, immunosuppressive therapy compromises the host immune response . The present study assessed the effects of immunosuppression by cyclosporine (CsA) or tacrolimus (Tac) on the indigenous bacterial flora of transplanted human skin . We found that a 6-day course of treatment with CsA or Tac was followed by an increased prevalence of bacterial isolates, mostly evidenced by a change in the spectrum of graft bacterial flora from Staphylococcus aureus and coagulase-negative staphylococci toward more pathogenic strains such as Escherichia coli, Enterococcus faecium, micrococcus, and pseudomonas . The mouse skin adjacent to the graft remained sterile, precluding the possibility of graft contamination with mouse flora. Transplant Proc, 2003 Sep, 35(6), 2304 - 6 Resistance of gram-positive pathogens to antibiotics is a therapeutic challenge after liver transplantation: clinical experience in one center with linezolid; Odakowska-Jedynak U et al.; BACKGROUND: Orthotopic liver transplantation has become an established therapeutic option for a large variety of fulminant and chronic liver diseases . Postoperative infections are the major cause of morbidity and the leading cause of mortality . The microbes responsible for these severe infections are predominantly gram-positive . METHODS: This article reviews results of linezolid therapy based on the clinical characteristics, microbial features, and outcomes of severe bacterial infections due to known or suspected resistant gram-positive species in selected liver allograft recipients . RESULTS: Among the 7 patients who received linezolid, methacillin-resistant Staphylococcus aureus . was isolated from 3, no pathogen from 2 patients, and serious pulmonary infection in 2 patients, 1 of whom had to be reintubated due to of respiratory failure . Cholangitis observed in 5 of 7 patients was caused by enterococci and staphylococci with septicemia in 1 subject . All patients demonstrated clinical improvement; microbiological eradication was observed in 4 patients . The majority of reported adverse events were mild or moderate in intensity . No potential drug interactions were observed between linezolid and concomitant medication . CONCLUSIONS: In the present study, linezolid proved to be effective and well tolerated . In summary, linezolid may represent an effective and safe antimicrobial agent for the treatment of infections due to susceptible and resistant gram-postive bacteria after solid organ transplantation. Syst Appl Microbiol, 2003 Sep, 26(3), 423 - 33 Combining denaturing gradient gel electrophoresis of 16S rDNA V3 region and 16S-23S rDNA spacer region polymorphism analyses for the identification of staphylococci from Italian fermented sausages; Blaiotta G et al.; Separation of amplified V3 region from 16S rDNA by denaturing gradient gel electrophoresis (PCR-DGGE) and 16S-23S rDNA intergenic spacer region polymorphism (ISR-PCR) analyses were tested as tool for differentiation of staphylococcal strains commonly isolated from fermented sausages . Variable V3 regions of 25 staphylococcal reference strains and 96 wild strains of species belonging to the genera Staphylococcus, Micrococcus and Kocuria were analyzed . PCR-DGGE profiles obtained were species-specific for S . sciuri, S . haemolyticus, S . hominis, S . auricularis, S . condimenti, S . kloosi, S . vitulus, S . succinus, S . pasteuri, S . capitis and S . (Macrococcus) caseolyticus . Moreover, 7 groups could be distinguished gathering the remaining species as result of the separation of the V3 rDNA amplicons in DGGE . Furthermore, the combination of the results obtained by PCR-DGGE and ISR-PCR analyses allowed a clear differentiation of all the staphylococcal species analysed, with exception of the pairs S . equorum-S . cohnii and S . carnosus-S . schleiferi . The suitability of both molecular techniques and of the combination their results for the identification of staphylococci was validated analysing partial nucleotide sequence of the 16S rDNA of a representative number of wild strains. Probl Tuberk Bolezn Legk, 2003, (8), 40 - 3 {Specific changes in the phagocytic activity of leukocytes in children after BCG vaccination}; Sanakoeva LP; In 185 children vaccinated with BCG, the phagocytic activity of leukocytes (PAL) was studied in different postimmunization periods . The modified procedure for examining PAL that reveals and assays specific phagocytic changes irrespective of the level of PAL was used . A suspended matter mixture of BCG mycobacteria (a specific object) and white saprophytic staphylococci (a non-specific object) was used as the objects of phagocytosis . A specific increase in PAL in the first 1.5 years after vaccination and the specific changes of phagocytosis during a vaccination process were first established . There was a close relationship of the specific changes of phagocytosis to the size of an infiltrate at the site of vaccination injection . The strain of antituberculosis immunity may be more objectively assessed by the phagocytic test than by the size of a skin scar and by the Mantoux test with 2TE PPD-L . The phagocytic test is recommended for individual monitoring of a antituberculosis vaccination process in children. Ophthalmology, 2003 Oct, 110(10), 1946 - 51 Antibiotic resistance patterns of ocular bacterial flora: a prospective study of patients undergoing anterior segment surgery; Ta CN et al.; PURPOSE: To determine the antibiotic susceptibility patterns of conjunctival bacterial flora isolated preoperatively from patients undergoing anterior segment surgery . DESIGN: Prospective observational study . PARTICIPANTS: One hundred fifty-six eyes from 139 patients scheduled for anterior segment surgery were enrolled over a 6-month period from August 2001 to February 2002 . METHODS: Conjunctival cultures were obtained on the day of surgery before povidone-iodine or antibiotic application . MAIN OUTCOME MEASURES: Bacterial isolates were identified and tested for antibiotic susceptibility using the Kirby-Bauer disc-diffusion technique . RESULTS: Among the 156 eyes studied, 36 were from patients who had undergone either bilateral surgery or more than one surgery in the same eye . Only the first eyes of the 120 patients that underwent initial ocular surgery were included in our analysis . Of these 120 eyes, 21 (18%) showed no bacterial growth . Of the 143 bacterial strains isolated from the remaining 99 eyes, 112 (78%) were coagulase-negative staphylococci (CNS) . Among the CNS, greater than 90% were susceptible to cefotaxime, levofloxacin, imipenem, meropenem, vancomycin, and each of the aminoglycosides except neomycin . Between 70% and 90% of the CNS were susceptible to cefazolin, neomycin, ciprofloxacin, ofloxacin, norfloxacin, and chloramphenicol . Less than 70% of the isolated CNS were sensitive to the penicillin analogues, ceftazidime, erythromycin, and tetracycline . CONCLUSIONS: Preoperative conjunctival isolates of CNS seem to be most sensitive to vancomycin, the aminoglycosides (except neomycin), and levofloxacin. Pediatr Infect Dis J, 2003 Sep, 22(9 Suppl), S158 - 63 Linezolid versus vancomycin in the treatment of known or suspected resistant gram-positive infections in neonates; Deville JG et al.; BACKGROUND: Gram-positive infections caused by susceptible and resistant strains of Staphylococcus aureus, coagulase-negative staphylococci and enterococci are increasing problems in neonates . Linezolid, a new oxazolidinone, is active against these pathogens and has recently been approved by the Food and Drug Administration for treating Gram-positive infections in pediatric patients . OBJECTIVE: To compare the clinical efficacy and safety of intravenous and oral linezolid with vancomycin (10 to 15 mg/kg every 6 to 24 h) in neonates (age 0 to 90 days) . METHODS: Hospitalized infants with known or suspected hospital-acquired pneumonia, complicated skin or skin structure infections, bacteremia or other infections (e.g . pyelonephritis, abdominal abscess) were eligible . Test-of-cure clinical response was evaluated at follow-up . RESULTS: Sixty-three neonates, randomized 2:1 to linezolid (n = 43) or vancomycin (n = 20) were included in the intent-to-treat group . Clinical cure rates at follow-up in the intent-to-treat group were higher, but not significantly different, for linezolid vs . vancomycin (78% vs . 61%; P = 0.196) . Corresponding cure rates in clinically evaluable patients were 84% vs . 77% (P = 0.553) for linezolid and vancomycin, respectively . Pathogen eradication rates were as follows in the linezolid and vancomycin groups, respectively: S . aureus (67% vs . 60%; P = 0.850); coagulase-negative staphylococci (88% vs . 100%; P = 0.379); and enterococci (71% vs . 0%; P = 0.168) . Results for hematology and chemistry assays were similar between treatment groups . Fewer linezolid-treated neonates had drug-related adverse events than vancomycin-treated neonates (12% vs . 32%; P = 0.058) . CONCLUSIONS: Linezolid is well-tolerated and as effective as vancomycin in the treatment of resistant Gram-positive infections in neonates. J Antimicrob Chemother, 2003 Nov, 52(5), 820 - 5 Epub 2003 Sep 30. Bacteriological outcome of combination versus single-agent treatment for staphylococcal endocarditis; Drinkovic D et al.; OBJECTIVE: To analyse the bacteriological outcome of combination versus single-agent antimicrobial treatment in staphylococcal endocarditis . PATIENTS AND METHODS: Retrospective review of 152 episodes: 91 cases of native valve endocarditis (NVE), 74 due to Staphylococcus aureus and 17 due to coagulase-negative staphylococci (CoNS); and 61 cases of prosthetic valve endocarditis (PVE), 29 due to S . aureus and 32 due to CoNS . RESULTS: Valves from patients with S . aureus NVE treated with any kind of combination antibiotic treatment were no more likely to be culture-negative than those treated with a single agent {19 (45%) of 42 versus 13 (41%) of 32; P = 0.69} . This finding remained unchanged when cases of CoNS NVE were added to the S . aureus group . In PVE, after adjusting for duration of treatment, valves from patients receiving any kind of combination treatment were 5.9 times (95% confidence interval 1.3-27.5) more likely to be culture-negative than those receiving monotherapy (P = 0.024) . Patients treated for >14 days were more likely to be culture-negative than those treated for <or=14 days {49 (83%) of 59 versus 29 (31%) of 93; P < 0.001} . CONCLUSIONS: In staphylococcal NVE, combination treatment is not superior to monotherapy in sterilizing infected valves, but in PVE combination treatment confers an advantage. J Clin Pathol, 2003 Oct, 56(10), 782 - 5 Fast, simultaneous, and sensitive detection of staphylococci; Stuhlmeier R et al.; AIMS: Bacterial infections are common and are involved in many forms of disease, ranging from arthritis to food poisoning . Of much concern are nosocomial infections, especially the increasing resistance of bacteria to methicillin . A prerequisite for the successful treatment of bacterial infections is a specific and sensitive method of detecting microorganisms . METHODS: Some methods to detect bacteria are time consuming, whereas others are faster but lack specificity and/or sensitivity . This article describes an optimised polymerase chain reaction (PCR) method that enables the simultaneous detection of different bacteria . A prerequisite for sensitive PCR is a method to isolate and recover extremely small amounts of bacterial DNA . This study used a new method to isolate DNA and compared the results to an established method . RESULTS: The method could detect fewer than 10 Staphylococcus aureus, methicillin resistant S aureus, Staphylococcus epidermitis, and other bacteria and it took less than two hours to perform . CONCLUSION: The rapid DNA isolation method used in conjunction with the optimised PCR makes it possible to confirm the presence or absence of extremely small numbers of bacteria . Using real time PCR would shorten the procedure even further . This method might therefore contribute to more timely and specific interventions. APMIS, 2003 Sep, 111(9), 905 - 14 Evaluation of different disk diffusion/media combinations for detection of methicillin resistance in Staphylococcus aureus and coagulase-negative staphylococci; Skov R et al.; In order to find a disk diffusion method with both high sensitivity and specificity for determination of methicillin resistance primarily for S . aureus but also for coagulase-negative staphylococci we screened several methodological variants using a material of 66 S . aureus comprising of 11 methicillin-susceptible, 18 borderline-resistant, and 37 methicillin-resistant strains . Only four of the combinations studied performed with both high sensitivity and specificity . Two of these, the Columbia agar +4.5% NaCl and Mueller Hinton agar +2% NaCl combined with a 5 microg oxacillin disk, confluent inoculum and 24 h incubation at 35 degrees C were further evaluated using 105 MRSA and 91 mecA-negative S . aureus and 193 clinical isolates of coagulase-negative staphylococci . The Columbia agar +4.5% NaCl performed excellently for both S . aureus and coagulase-negative staphylococci . For Columbia agar +4.5% NaCl using a 5 microg oxacillin disk we suggest an interpretive zone diameter of R < or =15 mm and S > or =16 mm for S . aureus and R < or =24 mm and S >or =26 mm for coagulase-negative staphylococci . The Mueller Hinton agar +2% NaCl performed well for coagulase-negative staphylococci but for S . aureus at least three (3%) very major errors were found, making this method less attractive. Curr Opin Crit Care, 2003 Oct, 9(5), 403 - 12 Antimicrobial resistance among gram-positive organisms in the intensive care unit; Clark NM et al.; PURPOSE OF REVIEW: The epidemiology of gram-positive pathogens in the intensive care unit are reviewed, recent trends in antimicrobial resistance among these organisms are discussed, and the significance of these data with respect to treatment are considered . RECENT FINDINGS: Results of surveillance studies published in 2001 and 2002 have demonstrated that gram-positive organisms such as Staphylococcus aureus, coagulase-negative staphylococci, and enterococci are among the most common bacteria infecting patients in intensive care units . Furthermore, these organisms are becoming increasingly resistant to available antimicrobial agents, and 2002 has ushered in worrisome developments such as the appearance of vancomycin-resistant S . aureus . Community-acquired methicillin-resistant S . aureus and the rise in incidence of vancomycin-resistant enterococci are other problems of great concern . Novel antibiotics such as quinupristin/dalfopristin and linezolid have activity against these agents, but resistance may develop to these agents as well . Studies have shown that infections caused by antibiotic-resistant organisms may be associated with increased morbidity, mortality, and costs . Exposure to antibiotics is a major risk factor for producing antibiotic resistance in patients, and methods to limit the spread of these organisms include restriction of antibiotic use, infection control, surveillance programs, and isolation procedures . SUMMARY: An awareness of the prevalence and patterns of resistance among gram-positive nosocomial pathogens is vital for the appropriate treatment of hospitalized patients . In addition, efforts must be made to minimize the selection and spread of these organisms. Antimicrob Agents Chemother, 2003 Oct, 47(10), 3046 - 52 Novel plasmid-borne gene qacJ mediates resistance to quaternary ammonium compounds in equine Staphylococcus aureus, Staphylococcus simulans, and Staphylococcus intermedius; Bjorland J et al.; We identified a novel plasmid-borne gene (designated qacJ) encoding resistance to quaternary ammonium compounds (QACs) in three staphylococcal species associated with chronic infections in four horses . qacJ was located on a 2,650-bp plasmid (designated pNVH01), a new member of the pC194 family of rolling-circle replication plasmids . The 107-amino-acid protein, QacJ, showed similarities to known proteins of the small multidrug resistance family: Smr/QacC (72.5%), QacG (82.6%), and QacH (73.4%) . The benzalkonium chloride MIC for a qacJ-containing recombinant was higher than those for otherwise isogenic recombinants expressing Smr, QacG, or QacH . Molecular epidemiological analyses by pulsed-field gel electrophoresis suggested both the clonal spread of a qacJ-harboring Staphylococcus aureus strain and the horizontal transfer of pNVH01 within and between different equine staphylococcal species . The presence of pNVH01 of identical nucleotide sequence in different staphylococcal species suggests that recent transfer has occurred . In three of the horses, a skin preparation containing cetyltrimethylammonium bromide had been used extensively for several years; this might explain the selection of staphylococci harboring the novel QAC resistance gene. Support Care Cancer, 2003 Dec, 11(12), 790 - 4 Epub 2003 Sep 20. Peripherally inserted central venous catheters for autologous blood progenitor cell transplantation in patients with haematological malignancies; Harter C et al.; BACKGROUND: In contrast to the high risk of haemorrhage associated with the implantation of a central venous catheter (CVC) via the internal jugular or subclavian access, the use of a peripherally inserted catheter (PICC) offers the advantage of a lower risk of bleeding complications . However, the rate of phlebitis is higher with the PICC and its use has been declining . We have studied the benefits and adverse events of a new type of PICC and a common type . METHODS: From October 1999 to October 2001, 70 PICCs (Olimpicc, Vygon, Germany, n=40; and LIFECATH-PICC(PUR)5FR Vygon, Germany, n=30) were inserted into 66 patients with haematological malignancies and used for high-dose chemotherapy, total parenteral nutrition and autologous blood stem cell transplantation . While removing the catheter, central and peripheral blood cultures were taken . The catheter tip was investigated by the semi-quantitative roll-out method of Maki . RESULTS: Sixty-five PICCs were removed after a median of 8.9 days . In five cases a catheter-associated significant colonisation with coagulase-negative staphylococci occurred . In two instances catheter-related bacteraemia was found . CONCLUSION: In our study this catheter system was inserted in 94% of patients without problems and showed a low incidence of phlebitis (5/65) . Because of the high rate of catheter malfunction reported during and after our study, the Olimpicc catheter is no longer available . The PICC system, and the LIFECATH-PICC(PUR)5FR in particular, offers a safe and effective alternative for central venous access to the internal jugular vein. J Food Prot, 2003 Sep, 66(9), 1693 - 6 Enterotoxin production by Staphylococcus aureus isolated from mastitic cows; Cenci-Goga BT et al.; Staphylococcus aureus is an important cause of mastitis in cows . The ability of S . aureus strains to produce one or more enterotoxins in milk and dairy products is linked to staphylococcal food poisoning . To determine whether staphylococci causing bovine mastitis could cause human foodborne intoxication, the production of staphylococcal enterotoxins A through D (SEA, SEB, SEC, and SED) by 160 S . aureus isolates was evaluated with the use of a reverse passive latex agglutination enterotoxin kit . All S . aureus strains were isolated over a 9-month period from 2,343 routine submissions of a composite quarter collection of individual mastitic cows at 18 dairy farms in the San Joaquin Valley in California . Prior to enterotoxin detection, isolates were grown by a method that enhances the in vitro synthesis of enterotoxin . Twenty-two of 160 S . aureus isolates produced enterotoxin . Seven produced SEC, 12 produced SED, and 3 produced both SEC and SED . None of the isolates produced SEA or SEB. J Microbiol Methods, 2003 Oct, 55(1), 279 - 86 Development of specific PCR primers for a rapid and accurate identification of Staphylococcus xylosus, a species used in food fermentation; Morot-Bizot S et al.; Twenty-seven Staphylococcus strains isolated from food and food environments were assigned to Staphylococcus xylosus by API-Staph system . But only seven isolates had similar patterns to this species when compared to the pulse-field gel electrophoresis patterns of 12 S . xylosus strains . To perform a rapid identification of the S . xylosus species, a random amplified polymorphic DNA product of 539-bp shared by all of the S . xylosus strains was used to design a pair of primers . These primers were species-specific for S . xylosus when tested by PCR on 21 staphylococci species . This specific PCR assay confirms the identification of the seven isolates identified by PFGE to S . xylosus . In conclusion, we developed specific PCR primers for a rapid and accurate identification of the S . xylosus species. Ann Hematol, 2003 Oct, 82 Suppl 2, S105 - 17 Epub 2003 Sep 09. Antimicrobial therapy of unexplained fever in neutropenic patients--guidelines of the Infectious Diseases Working Party (AGIHO) of the German Society of Hematology and Oncology (DGHO), Study Group Interventional Therapy of Unexplained Fever, Arbeitsgemeinschaft Supportivmassnahmen in der Onkologie (ASO) of the Deutsche Krebsgesellschaft (DKG-German Cancer Society); Link H et al.; Cytostatic chemotherapy of hematological malignancies is often complicated by neutropenia, which increases the risk of infections, especially if the neutrophil count is below 500/microl . Frequently, fever is the first, and in most patients the only, sign of an infection . Unexplained fever is defined as follows: temperature of >/=38.3 degrees C or >/=38.0 degrees C for at least 1 h, or measured twice within 12 h, if the neutrophil count is <500/microl or <1000/microl with predicted decline to 500/microl . Different risk categories can be identified according to the duration of neutropenia: low risk </=5 days, intermediate risk 6-9 days, high risk >/=10 days . An empirical mono- or duotherapy with antipseudomonal and antistreptococcal agents should be initiated immediately . In the low risk patient group, oral therapy with cipro-, levo-, or ofloxacin combined with amoxicillin/clavulanic acid is permissible . For standard and high risk patients, monotherapy can be carried out with either ceftazidime, cefepime, piperacillin with tazobactam or a carbapenem . In duotherapy, a single dose of an aminoglycoside is combined with acylaminopenicillin or a cephalosporin of the third or fourth generation . The addition of glycopeptides in empirical therapy should only be considered in the presence of severe mucositis, or if a catheter-associated infection is suspected . If fever persists after 72-96 h of first-line therapy with antibiotics, the regimen should be modified (with the exception of e.g . coagulase-negative staphylococci infections, because these infections take longer to respond) . Intermediate risk patients should additionally receive an aminoglycoside after monotherapy (penicillin or a cephalosporin) . If a carbapenem was administered for monotherapy, this can be followed by a quinolone and/or a glycopeptide . In the high risk group, the same modifications should be made as in the intermediate risk group but with additional systemic antifungal treatment . In the presence of unexplained fever, fluconazole can be administered at first, but if this fails, amphotericin B (conventional or liposomal), itraconazole, voriconazole or caspofungin should be started . After defervescence to <38 degrees C, treatment should be continued for 7 days if the neutrophil count is <1000/microl, and for 2 days if the neutrophil count is >1000/microl. Ann Hematol, 2003 Oct, 82 Suppl 2, S149 - 57 Epub 2003 Sep 09. Central venous catheter (CVC)-related infections in neutropenic patients--guidelines of the Infectious Diseases Working Party (AGIHO) of the German Society of Hematology and Oncology (DGHO); Fatkenheuer G et al.; Catheter-related infections cause considerable morbidity in hospitalised patients . The incidence does not seem to be higher in neutropenic patients than in non- neutropenic patients . Gram-positive bacteria (coagulase-negative staphylococci, Staphylococcus aureus) are the most frequently cultured pathogens, followed by Candida species . In contrast, Gram-negative bacteria play only a minor role in catheter-related infections . Positive blood cultures are the cornerstone in the diagnosis of catheter-related infections, while local signs of infection are only rarely present . However, a definite diagnosis generally requires the removal of the catheter and its microbiological examination . The role plate method with semiquantitative cultures (Maki) has been established as standard in most laboratories . Other standard procedures use quantitative techniques (Sherertz, Brun-Buisson) and are more sensitive . For therapy of catheter-related infections, antibiotics are administered according to the susceptibility of the cultured organism . Routine administration of gylcopepticed antibiotics is not indicated . Removal of the catheter has to be considered in any case of suspected catheter-related infection and is obligatory in Staphylococcus aureus and Candida infections . Tunnel or pocket infection of long-term catheters is always an indication for removal . In the future, the rate of catheter-related infections in neutropenic patients may be reduced by the use of catheters coated with antimicrobial agents. Int J Antimicrob Agents, 2003 Sep, 22(3), 228 - 36 Msr(A) and related macrolide/streptogramin resistance determinants: incomplete transporters? Reynolds E, Ross JI, Cove JH. The gene msr(A) confers inducible resistance to 14-membered-ring macrolides and type B streptogramins (MS(B) resistance) in staphylococci . The encoded hydrophilic protein (Msr(A)) is 488 amino acids and contains two ATP-binding motifs characteristic of the ABC transporters . The classical organisation of ABC transporters requires interaction between the two cytoplasmically located ATP-binding domains with two hydrophobic domains positioned in the membrane . Msr(A) appears to mediate drug efflux and yet contains no hydrophobic membrane spanning domains . In addition, Msr(A) functions in previously sensitive heterologous hosts such as Staphylococcus aureus in the absence of other plasmid encoded products . Current research on Msr(A) and related determinants in Gram-positive cocci and in antibiotic producing organisms is reviewed . Alternative hypotheses for the mechanism of action of Msr(A) (i.e . active transport vs . ribosomal protection) are discussed . Evidence indicating Msr(A) may have a role in virulence in addition to conferring antibiotic resistance is also considered. Int J Syst Evol Microbiol, 2003 Sep, 53(Pt 5), 1647 - 54 Macrococcus brunensis sp . nov., Macrococcus hajekii sp . nov . and Macrococcus lamae sp . nov., from the skin of llamas; Mannerova S et al.; Eight strains of Gram-positive, catalase- and oxidase-positive cocci were isolated from the skin of llamas (Lama glama L.) and characterized using a polyphasic approach . These strains were assigned to the genus Macrococcus on the basis of their phenotypic properties (resistance to bacitracin and sensitivity to furazolidone) and DNA base content (40-42 mol% G+C) . Phylogenetic analysis based on 16S rDNA confirmed that the strains are members of the genus MACROCOCCUS: They differed from all hitherto described macrococcal species in their production of phosphatase and reduction of nitrate (most strains) and the inability to produce acid from glycerol or to grow in 7.5 % NaCl . Ribotyping (EcoRI), macrorestriction analysis (XbaI) and fatty acid methyl ester analysis divided the strains from llamas into three stable clusters . Moreover, ribotyping differentiated the strains analysed not only from previously described macrococcal species but also from oxidase-positive staphylococci . DNA-DNA hybridization confirmed that the three clusters represent separate genomic groups (similarity values<54 %) . All the results showed that the strains represent three novel species, for which the names Macrococcus hajekii sp . nov . (type strain CCM 4809(T)=LMG 21711(T)), Macrococcus brunensis sp . nov . (type strain CCM 4811(T)=LMG 21712(T)) and Macrococcus lamae sp . nov . (type strain CCM 4815(T)=LMG 21713(T)) are proposed. FEMS Immunol Med Microbiol, 2003 Sep 22, 38(2), 153 - 8 Strict infection control measures do not prevent clonal spread of coagulase negative staphylococci colonizing central venous catheters in neutropenic hemato-oncologic patients; van Pelt C et al.; Coagulase negative staphylococci (CoNS) are a main cause of catheter related infections (CRI) . Earlier studies (1994-1996) revealed a high incidence of CRI (6 per 1000 catheter days) among neutropenic hemato-oncologic patients in the Erasmus MC Hematology Department (Rotterdam, The Netherlands) . This was mainly explained by expansion of two methicillin resistant Staphylococcus epidermidis (MRSE) clones (Nouwen et al., J . Clin . Microbiol . 36 (1998) 2696-2702) . In a new, 16-bed unit in the same institution, we investigated the effect of strict clinical isolation measures on the incidence of CRI . During two 6-month screening periods (period I: April 1998-December 1998 and period II: April 1999-October 1999) all patients receiving a central venous catheter were prospectively monitored for the development of CRI . During period I every visitor of the cubicles had to wear hair caps, masks, gowns and gloves . During period II these procedures were abolished, but hands were cleansed using alcohol and masks were worn during both periods in case of coughing and sneezing . All CoNS strains isolated from blood cultures were genetically classifies by pulsed field gel electrophoresis (PFGE) . The incidence of CRI during period I was 13.0 per 1000 catheter days, in comparison to 9.6 in period II (P=0.84) . During this latter period, 19 CRI were diagnosed, 14 catheter related bacteremia episodes (CRB) and five local infections . Seventy-two percent (n=9) of CRB were due to a CoNS . The mean catheter survival until appearance of a CRI increased from 43 days during period I to 78 days in period II (P=0.39) . The mean catheter survival until infection related removal was increased from 43 days to 133 days (P=0.12) . During period I less experienced intervention radiologists introduced the catheters, which may have limited the efficacy of the strict hygiene measures . Thus, abolishing strict isolation precautions had no negative effect on the incidence of CRI . After genotyping of 38 MRSE strains isolated from blood and central venous catheter cultures of 12 patients in period II, eight PFGE types were found . Three types were found in more than one patient, but based on epidemiological data patient-to-patient spread could not be proven . No genotypic identity between patient and personnel CoNS isolates was shown and the two major clonal types that were present between 1994 and 1996 were not encountered . However, from December 1998 onwards new MRSE clones could be identified (types E and J) . In conclusion, despite a constant rate of CRI and implementation of optimal patient care, clonal spread of MRSE strains was not prevented by strict hygiene measures. J Clin Oncol, 2003 Sep 15, 21(18), 3520 - 5 Catheter design influences recurrence of catheter-related bloodstream infection in children with cancer; Flynn PM et al.; PURPOSE: Multiple studies have demonstrated that catheter-related bloodstream infections (CRBI) can be successfully treated without catheter removal (in situ therapy), but there is insufficient information available to determine if catheter design can influence the eradication of bacteremia or recurrence . PATIENTS AND METHODS: Bacteremic episodes in patients at St Jude Children's Research Hospital between January 1996 and May 2001 were identified and patient records were reviewed . RESULTS: A total of 172 unique episodes of CRBI were identified . In situ therapy resulted in successful eradication of bacteremia in 87% of the episodes . Bacteremia recurred in 10% of the episodes . Although catheter design (Hickman and Broviac versus totally implantable central venous catheter) did not influence short-term eradication of bacteremia, totally implantable central venous catheters were significantly associated with recurrence of bacteremia (odds ratio, 10; 95% confidence interval, 3.1 to 33.3) . In a multivariable analysis, this association between catheter design and recurrence remained statistically significant after adjustment for other factors that influenced recurrence in this study (isolation of coagulase-negative staphylococci and inadequate duration of initial antibiotic therapy) . CONCLUSION: This study demonstrates that patients with CRBI with a totally implantable central venous catheter in place are more likely to develop recurrent bacteremia . Management strategies to prevent recurrence in this setting should be explored. Diagn Microbiol Infect Dis, 2003 Sep, 47(1), 303 - 11 Detection of antimicrobial resistance by small rural hospital microbiology laboratories: comparison of survey responses with current NCCLS laboratory standards; Stevenson KB et al.; Microbiology laboratory personnel from 77 rural hospitals in Idaho, Nevada, Utah, and eastern Washington were surveyed in July 2000 regarding their routine practices for detecting antimicrobial resistance . Their self-reported responses were compared to recommended laboratory practices . Most hospitals reported performing onsite bacterial identification and susceptibility testing . Many reported detecting targeted antimicrobial resistant organisms . While only 5/61 hospitals (8%) described using screening tests capable of detecting all 8 targeted types of resistance, most (57/61, 93%) were capable of accurately screening for at least 6 types . Conversely, most hospitals (58/61, 95%) reported confirmatory testing capable of identifying only 3 or fewer resistance types with high-level penicillin resistance among pneumococci, methicillin and vancomycin resistance among staphylococci and enterococci, and extended spectrum beta-lactamase production by Gram-negative bacilli presenting the greatest difficulties . Furthermore, only 50% of hospitals compiled annual antibiogram reports to help physicians choose initial therapy for suspected infectious illnesses . This survey suggests that the antimicrobial susceptibility testing in many rural hospitals may be unreliable. Zh Mikrobiol Epidemiol Immunobiol, 2003 Jul-Aug, (4), 51 - 5 {Characterization of relationships between pro- and eukaryotic cells under the conditions of intracellular symbiosis of Staphylococcus aureus in the mucosa cells of the respiratory and digestive systems organs in terms of ecology and morphology}; Stadnikov AA et al.; The ecological and morphological analysis of reorganization processes in the cell and tissue structures of the mucous membranes of the respiratory and digestive systems in rats under the conditions of their prolonged symbiotic interactions with staphylococci (in the body of rats as a whole, as well as on the model of organotypic cultures in diffusion chambers in vivo) was carried out with the use of light and electron microscopy, radioautography and morphometry . The morphological equivalents of phenotypic reactions in the cell elements of the mucous membranes of the respiratory and digestive systems (epitheliocytes, leimyocytes, endotheliocytes, macrophages and fibroblasts) under the conditions of staphylococcus persistence were revealed . Adaptive and reactive shifts in eukaryotic cells were manifested by an increased volume of nuclei and greater proportion of euchromatin and a decreased DNA-synthetic and proliferative activity . Microorganisms located inside the cells underwent ultrastructural reorganization. Zh Mikrobiol Epidemiol Immunobiol, 2003 Jul-Aug, (4), 47 - 51 {Properties of Staphylococcus aureus in case of the unfavorable course of burn infection}; Brudastov IuA et al.; Clinical and microbiological study of wound discharge from 35 patients demonstrated a relationship between biological properties of S . aureus, the causative agent of burn infection, and the course of the infected burn trauma . The prognostic importance of the antioxidant properties of these staphylococci was established: in cases of the unfavorable course of the burn process they showed essentially greater resistance to peroxinitrite and higher superoxide dismutase activity in comparison with the cultures isolated from patients with uncomplicated wound infection. Clin Orthop, 2003 Sep, (414), 89 - 94 PCR rapidly detects methicillin-resistant staphylococci periprosthetic infection; Tarkin IS et al.; Optimal treatment of methicillin-resistant staphylococcal periprosthetic infections is predicated on rapid and reliable detection of these organisms . Culture has served as the gold standard for identification of these organisms despite shortcomings with sensitivity and processing time . The objective of the current study was to investigate a polymerase chain reaction assay aimed at rapid genomic detection of methicillin-resistance in staphylococci (mecA gene) . The feasibility of the molecular approach first was validated using a septic arthritis model consisting of 73 synovial fluid samples inoculated with methicillin-resistant staphylococci and four negative controls . MecA polymerase chain reaction then was done on 35 clinical samples from 18 patients obtained at the time of revision arthroplasty . Results of the polymerase chain reaction were compared with culture . MecA polymerase chain reaction successfully predicted the presence of methicillin-resistant staphylococci in the septic arthritis model . In the clinical samples studied, the polymerase chain reaction results were concordant with culture results in 34 of the 35 samples tested . The one discordant result represented a false-positive culture result . The molecular assay was processed in less than 5 hours compared with 2 to 3 days for culture . Detection of methicillin-resistant staphylococci involved in periprosthetic infections by the polymerase chain reaction is a rapid and reliable approach. Braz J Infect Dis, 2003 Apr, 7(2), 142 - 8 Epub 2003 Nov 19. Comparison between the jugular and subclavian vein as insertion site for central venous catheters: microbiological aspects and risk factors for colonization and infection; Sadoyama G et al.; Bacterial counts were made of catheter insertion site and of catheter tips to help determine risk factors associated with catheterization of the jugular and subclavian veins . Among the 116 patients included in this study, 69% had central venous catheters (CVC) in the subclavian vein . Seven or more days catheterization (p=0.001) and > or =3 invasive devices (p=0.01) were infection risk factors associated with catheterization of the jugular vein . More than half of the patients presented high colony counts at the insertion site (> or =200 CFU/20 cm2) and 27% of the catheter tips were contaminated . The risk factors associated with contaminated catheter tips were > or =14 days hospital stay (p=0.02), > or =7 days catheterization (p=0.01) and antibiotic therapy (p=0.04) . Coagulase-negative staphylococci (CoNS) and Staphylococcus aureus were the most common microorganisms at the insertion site (78%) and in the catheter tip (94%) . Five patients presented sepsis (4.1%), four caused by Staphylococci and one by GNB . Twelve patients had the same microorganisms at the insertion site and catheter tip . We found a high prevalence of ORSA (62.5%) and ORCoNS (57.1%) in catheter tips . The high counts of staphylococci, including ORSA and ORCoNS, at the insertion site, and the significant association of this colonization with catheter tip contamination, indicate that the skin is an important reservoir of microorganisms associated with catheter-related bloodstream infection (CR-BSI) . Health professionals should be aware of this potential source of infection at the CVC insertion site. Microb Drug Resist, 2003 Fall, 9(3), 273 - 82 Application of molecular typing methods to characterize nosocomial coagulase-negative staphylococci collected in a Greek hospital during a three-year period (1998-2000); Spiliopoulou I et al.; A total of 143 methicillin-resistant coagulase-negative staphylococci (MR-CNS) collected between 1998 and 2000 at the University Hospital of Patras, Greece, were characterized by antibiogram and genomic typing to define the clonal types endemic in this hospital and their evolution during the 3-year period . These isolates corresponded to 93 methicillin-resistant Staphylococcus epidermidis (MRSE) and 50 other MR-CNS, which were isolated from patients in different wards, exclusively from blood and catheter tips cultures . Pulsed-field gel electrophoresis (PFGE) of SmaI macrofragments and hybridization of ClaI digests with mecA and murE DNA probes were performed . The application of these methodologies demonstrated the existence, persistence and spread of MRSE, MR-Staphylococcus haemolyticus, and MR-Staphylococcus hominis clones in this hospital, whereas the SmaI/murE hybridization pattern was shown to be a valuable tool for the MRSE identification. Pneumonol Alergol Pol, 2003, 71(1-2), 31 - 5 {Microorganisms isolated from clinical specimens of patients from the Department of Thoracic Surgery}; Czarniak E et al.; The clinical specimens received from patients hospitalized in Department of Thoracic Surgery between 1997 and 2001 were microbiologically examined . The main specimen for microbiological examination was pleural fluid (median 34%) . The frequency of specimens from bronchial tree increased significantly (from 4% to 26%) with concurrent decrease of sputum (from 29% to 6%) . Among isolated pathogens, Gram negative rods were the most frequent (median 48%) and Pseudomonas sp . was the main pathogen among them . Occurrence of staphylococci was median 22% and Staphylococcus aureus, with a little decrease in analyzed period, was still the main Gram positive pathogen . Simultaneously the occurrence of MRSA in the last three years dropped three times . The number of isolations of yeasts have risen from 5.8% to 10.3%. J Clin Microbiol, 2003 Sep, 41(9), 4324 - 7 Evaluation of three rapid methods for the direct identification of Staphylococcus aureus from positive blood cultures; Chapin K et al.; Staphylococci represent the most commonly encountered blood culture isolates . Differentiating Staphylococcus aureus from coagulase-negative staphylococci (CoNS) is important in guiding empirical therapy, especially since the majority of CoNS are contaminants . This study evaluated three rapid methods for the direct identification of S . aureus from blood cultures . A total of 157 patient blood cultures with gram stains showing gram-positive cocci in clusters were included . The following assays were evaluated: API RAPIDEC staph (API) (bioMerieux, Durham, N.C.), the tube coagulase test (TCT) read at 4 h, and peptide nucleic acid (PNA) fluorescence in situ hybridization (FISH) (AdvanDx, Woburn, Mass.) . All assays yielded results of S . aureus or non-S . aureus . The direct rapid results were compared to results obtained with isolated colonies using the AccuProbe Staphylococcus aureus Culture Identification Test (Gen-Probe, San Diego, Calif.) . API, TCT, and PNA FISH exhibited sensitivities of 96, 84, and 99% and specificities of 99, 100, and 100%, respectively . Direct identification testing by any of these three assays yielded acceptable performance and timely results . This ability to accurately detect S . aureus in blood cultures gives the physician information with which to initiate or tailor antimicrobial therapy . Coupled with direct susceptibility testing of positive blood culture broths, the patient and institution may experience improved outcomes. J Clin Microbiol, 2003 Sep, 41(9), 4259 - 63 Use of the VITEK 2 system for rapid identification of clinical isolates of Staphylococci from bloodstream infections; Spanu T et al.; Staphylococci are an increasing cause of bloodstream infections . Rapid reliable identification of these organisms is essential for accurate diagnosis and prompt effective treatment . We evaluated the ability of the VITEK 2 system (bioMerieux, Inc, Hazelwood, Mo.) to identify these organisms rapidly and accurately . A total of 405 clinically relevant nonduplicate staphylococcal isolates (Staphylococcus aureus, n = 130; coagulase-negative staphylococci, n = 275) collected from blood cultures were tested . VITEK 2 results were considered correct when they were identical to those furnished by the comparison method based on the ID 32 STAPH system (bioMerieux, Marcy l'Etoile, France) plus supplementary manual testing . When discrepancies occurred, isolate identity was verified by molecular typing . The VITEK 2 correctly identified 387 (95.6%) isolates at the species level: 379 (including all but one {99.2%} of 130 S . aureus isolates and 249 of 275 {90.5%} coagulase-negative isolates) were identified by the automated reading; for the other eight, supplemental tests suggested by the manufacturer had to be used . Only one strain (0.2%) was misidentified (Staphylococcus hominis as Staphylococcus epidermidis), and four (1%), all S . epidermidis, were not identified . For the remaining 13 strains (including 10 S . hominis), the VITEK 2 system was unable to discriminate among two species, and no supplemental tests were suggested for conclusive identification . Over 90% of results were obtained within 4 h . These results suggest that the VITEK 2 system can provide rapid, accurate, and reliable species-level identification of staphylococci responsible for bloodstream infections, although there is room for improvement in the identification of certain coagulase-negative species, especially S . hominis. Appl Environ Microbiol, 2003 Sep, 69(9), 5328 - 35 Fluorescence-activated cell sorting of specific affibody-displaying staphylococci; Wernerus H et al.; Efficient enrichment of staphylococcal cells displaying specific heterologous affinity ligands on their cell surfaces was demonstrated by using fluorescence-activated cell sorting . Using bacterial surface display of peptide or protein libraries for the purpose of combinatorial protein engineering has previously been investigated by using gram-negative bacteria . Here, the potential for using a gram-positive bacterium was evaluated by employing the well-established surface expression system for Staphylococcus carnosus . Staphylococcus aureus protein A domains with binding specificity to immunoglobulin G or engineered specificity for the G protein of human respiratory syncytial virus were expressed as surface display on S . carnosus cells . The surface accessibility and retained binding specificity of expressed proteins were demonstrated in whole-cell enzyme and flow cytometry assays . Also, affibody-expressing target cells could be sorted essentially quantitatively from a moderate excess of background cells in a single step by using a high-stringency sorting mode . Furthermore, in a simulated library selection experiment, a more-than-25,000-fold enrichment of target cells could be achieved through only two rounds of cell sorting and regrowth . The results obtained indicate that staphylococcal surface display of affibody libraries combined with fluoresence-activated cell sorting might indeed constitute an attractive alternative to existing technology platforms for affinity-based selections. J Vet Med Sci, 2003 Aug, 65(8), 899 - 906 Concentrations and specific antibodies to staphylococcal enterotoxin-C and toxic shock syndrome toxin-1 in bovine mammary gland secretions, and inflammatory response to the intramammary inoculation of these toxins; Kuroishi T et al.; To investigate the pathological role of staphylococcal enterotoxins (SEs) and toxic shock syndrome toxin-1 (TSST-1) in bovine mastitis, the production of SEs and TSST-1 was investigated in staphylococci isolated from 120 mammary gland secretions (MGS, 51 from no clinical sign-mammary glands and 69 from staphylococcal mastitic-mammary glands) collected from dairy farms where staphylococcal mastitis frequently occurred in Miyagi and Yamagata prefectures from 1997 to 1998 . Concentrations of these toxins and specific antibody titers in each MGS were also measured . Furthermore, SEC and TSST-1 were inoculated into lactating mammary glands and inflammatory responses were analyzed . A high percentage of staphylococci including Staphylococcus aureus and coagulase-negative staphylococci isolated from both no clinical sign- and mastitic-MGS produced both SEC and/or TSST-1 . The concentration of SEC increased with the severity of the mastitis, and was significantly higher (P<0.05) in acute mastitic-than in no clinical signs-MGS . Titers of specific antibodies to TSST-1 in MGS were significantly higher (P<0.05) than those to SEC, regardless of whether or not the cows were lactating or mastitic . Specific antibodies purified from MGS neutralized each toxin in vitro . A significant increase (P < 0.05) in somatic cell counts was induced by the intramammary inoculation of SEC but not TSST-1 . These findings indicated that SEC rather than TSST-1 plays an important role in the pathology of staphylococcal bovine mastitis . The inflammatory activity of TSST-1 was probably neutralized by specific antibodies in MGS. Yonsei Med J, 2003 Aug 30, 44(4), 571 - 8 Korean nationwide surveillance of antimicrobial resistance in 2000 with special reference to vancomycin resistance in enterococci, and expanded-spectrum cephalosporin and imipenem resistance in gram-negative bacilli; Lee KW et al.; Antimicrobial resistance surveillance is necessary to determine the size of the problem and to guide empirical selection of antimicrobial agents for treating infected patients . The aim of this study was to analyze the results of susceptibility tests performed by hospitals participating in the Korean Nationwide Surveillance of Antimicrobial Resistance (KONSAR) program . The rates of oxacillin-resistant staphylococci, penicillin-nonsusceptible pneumococci, and ampicillin-resistant E . faecium were over 70% . Ampicillin-resistant H . influenzae increased to 68% . Expanded-spectrum cephalosporin-resistant K . pneumoniae, fluoroquinolone-resistant E . coli, and imipenem-resistant P . aeruginosa remained at 16% through 27%, depending on the species . The proportions of vancomycin- resistant E . faecium and imipenem-resistant P . aeruginosa were 18 - 24% and 19-21%, respectively, indicating the seriousness of antimicrobial resistance . In conclusion, the increasing prevalence of resistant bacteria indicates that more concerted effort is required to conserve the usefulness of precious new antimicrobial agents. Pathol Biol (Paris), 2003 Sep, 51(7), 400 - 4 {In vitro activity of the pristinamycin against the isolated staphylococci in the french hospitals in 1999-2000}; Leclercq R et al.; One thousand six hundred and fifty clinically significant, consecutive and non redundant strains of staphylococci, including 863 Staphylococcus aureus and 787 coagulase negative staphylococci (CNS), were isolated between October 1999 and March 2000 in 35 French hospital laboratories . Susceptibilities were determined in each center by a standard diffusion method according to the recommendations of CA-SFM . Strains with vancomycin zone size diameter <17 mm were sent to the central laboratory for MIC determination of vancomycin by agar dilution, as recommended by the CA-CSFM . Frequencies of resistance to oxacillin were 38.6% for S . aureus (MRSA), 54% for the CNS, all species and 62% for S . epidermidis, respectively . The antibiotics tested showed a good activity against strains of S . aureus susceptible to oxacillin, more than 95% of strains being susceptible except for erythromycin (82.6%) . Against MRSA, vancomycin and prisitinamycin had the highest rates of susceptible strains, greater than 93% for the later antibiotic . More than 92% of strains of CNS susceptible or resistant to oxacillin were sensitive to pristinamycin . Pristinamycin displayed a good activity whether the strains were constitutively or inducibly resistant to MLS(B) . It comes out from this in vitro study that the rate of resistance of staphylococci to pristinamycin remains weak and stable in France . Pristinamycin is a good alternative for oral treatment of staphylococcal infections. Microb Pathog, 2003 Sep, 35(3), 119 - 24 Polarization of cytokine responses in B- and T-lymphocytes during Staphylococcus aureus infection; Gjertsson I et al.; The destructive course of Staphylococcus aureus arthritis is due to certain leukocytes and their products, mainly cytokines . The cellular source of cytokines mediating this inflammatory process has not been previously assessed on a protein level . Using a mouse model of hematogenously induced S . aureus infection the intracellular production of IFN-gamma, IL-4 and IL-10 in splenic B and T cells have been determined . This has enabled us to define distinct Th1 vs Th2 and Be1 vs Be2 populations of lymphocytes participating in S . aureus infection . Spleen cells were obtained before, and at different time intervals during the first week of infection and re-stimulated in vitro with staphylococcal peptidoglycan, formalin killed staphylococci and TSST-1 . The different antigens used for re-stimulation gave rise to different cytokine profiles in analysed T cells (identified as CD4+) and B cells (identified as CD19+) . TSST-1 acted as the most potent re-stimulator and we found that 40% of the CD4+ cells responded with IFN-gamma production, and unexpectedly almost 20% of the CD19+ cells . As to IL-4 and IL-10 production, the percentage of B cells expressing these cytokines was higher than the percentage T cells and the peak of their appearance appeared later than that of IFN-gamma . This finding indicates that Be1 cells are an important source of IFN-gamma early during the infection and that the production of the Th2 cytokines in B cells downregulates its production of IFN-gamma . In conclusion this study shows that both B and T cells contribute to the cytokine production during S . aureus infection in a complex pattern. J Hosp Infect, 2003 Aug, 54(4), 272 - 8 Genotypic and phenotypic properties of coagulase-negative staphylococci causing dialysis catheter-related sepsis; Spare MK et al.; Sixty coagulase-negative staphylococcus (CNS) isolates were recovered from the blood cultures or peritoneal dialysate effluent of 43 patients on renal dialysis . The patients had either renal dialysis catheter-related sepsis (CRS) or continuous ambulatory peritoneal dialysis (CAPD)-associated peritonitis . Isolates were characterized by biotyping, and genotyped by pulsed-field gel electrophoresis (PFGE) . Phenotypic properties of the strains were also investigated . Several genotypes were identified with no one specific strain of CNS being associated with CRS . However, closely related strains were isolated from several patients within the units studied, suggesting horizontal transfer of micro-organisms . Genotypic macro-restriction profiles did not concur with phenotypic profiles or biotypes, confirming that genotyping is required for epidemiological studies . All staphylococcal strains were investigated for the production of phenotypic characteristics . Significant differences were predominantly seen in the production of lipase, esterase and elastase in strains isolated from the renal patients with CRS and CAPD-associated peritonitis, compared with a non-septic control group . These phenotypic characteristics may therefore have a role in the maintenance of CRS in renal patients. Dtsch Tierarztl Wochenschr, 2003 Jul, 110(7), 295 - 8 Crowding and winter emergency feeding as predisposing factors for kerato-conjunctivitis in semi-domesticated reindeer in Norway; Aschfalk A et al.; Due to the hard environmental and climatic situation in late winter 1999, a herd of about 200 free-ranging, semi-domesticated reindeer was gathered in a paddock in northern Norway for emergency feeding . About the same number of reindeer was not corralled but supplementary fed on their winter pastures . The fodder was of relatively good quality but very dusty and fed in a very dry environment . Six weeks later, an outbreak of eye-infection was diagnosed in one third of the corralled reindeer; mild symptoms were observed in most of them, but 11 animals showed severe signs of disease . No signs of disease were found in the non-corralled animals . Ten reindeer died through emaciation, the eleventh was sacrificed . Histopathological diagnosis of two severely affected eyes revealed a severe purulent kerato-conjunctivitis with bacteria and plant particles embedded in purulent exudates on the cornea and conjunctiva . In one eye from the two most affected animals Actinomyces pyogenes, coagulase-negative Staphylococci and Escherichia coli and in the other one Staphylococcus aureus and Escherichia coli were found . The bacteria encountered in this study are not considered the primary cause of disease . They seem rather to be opportunistic infectious agents of eyes that have been irritated mechanically through exposure to dusty fodder in a dry environment . The stress through unfamiliar corralling of the reindeer, that followed an insufficient fodder supply, could be considered as an additional infection supporting factor . This case-report emphasises on the importance of different factors involved in favouring outbreaks of disease in reindeer, under intensified husbandry conditions . Even though crowding and emergency feeding may be, at certain circumstances, the only means of survival for reindeer, a negative impact of implied crowding diseases on their productivity, must be considered, as well. Med Dosw Mikrobiol, 2003, 55(1), 1 - 10 {Therapeutic effect of some antibiotics on experimental staphylococcal infection and its correlation with in vitro activity of antibiotics in sub-inhibitory concentration against Staphylococcus aureus strains}; Kaczmarska L et al.; The aim of the study was to demonstrate of whether the therapeutic effects of antibiotics depend on their in vitro activity in sub-inhibitory concentrations against staphylococci . Cloxacillin, gentamicin and lincomycin were used in the study . Groups of S . aureus strains, containing 6 strains with similar MIC values each but different sensitivity to sub-inhibitory antibiotic concentrations (sub-MIC) were selected (a total of 36 trains): i . strains increasing their sensitivity to phagocytosis and bactericidal activity of rabbit leukocytes after incubation with an antibiotic in 0.1 MIC concentration, ii . strains with sensitivity to the above factors unaffected by incubation with an antibiotic in 0.5 MIC concentration . The doses of staphylococci causing death of 90-100% of Swiss albino mice 10 days after i.p . infection were determined . The injected doses (LD 90-100) and various doses of antibiotics were used to determine ED50 values as well as the survival rate of the mice with experimental staphylococcal infections after treatment with these antibiotics . It was demonstrated that effective doses (ED 50) of the antiboitics were significantly lower when the antibiotics were administered once to mice infected with strains S . aureus sensitive to sub-MIC concentrations of the investigated antibiotics than for mice infected with strains resistant to their sub-MIC concentrations . Similar correlations were observed in mice which were given the antibiotics several times (for 7 days): the percentage of the surviving mice was higher in the group infected with sub-MIC sensitive strains . The therapeutic effect of cloxacillin, gentamicin and lincomycin demonstrated a significant correlation with the S . aureus strains used to induce the infections and their sensitivity, or lack of sensitivity in vitro, to phagocytosis and bactericdal activity of leukocytes in the presence of antibiotics in sub-MIC concentrations. J Dairy Sci, 2003 Jul, 86(7), 2382 - 9 Elevated milk soluble CD14 in bovine mammary glands challenged with Escherichia coli lipopolysaccharide; Lee JW et al.; The purpose of this study was to determine whether soluble CD14 (sCD14) in milk was affected by stage of lactation, milk somatic cell count (SCC), presence of bacteria, or lipopolysaccharide (LPS)-induced inflammation . Milk samples from 100 lactating cows (396 functional quarters) were assayed for sCD14 in milk to determine effects of stage of lactation, SCC, and intramammary infection . The concentration of sCD14 was highest in transitional milk (0 to 4 d postpartum) and in milk with high SCC (> 750,000 cells/ml) . Most of the infected quarters (> 80%) were infected by coagulase-negative staphylococci and yeast . No difference was found between noninfected and infected quarters . One quarter of six healthy lactating cows was challenged with 100 microg LPS in order to study the kinetics of sCD14 during an LPS-induced inflammation . Milk samples were collected at various intervals until 72 h after injection . Rectal temperature, milk tumor necrosis factor-alpha, and interleukin-8 increased immediately after challenge . The increase in sCD14 paralleled the increase in SCC, peaked at 12 h, and started to decline after 24 h . Serum leakage, as characterized by the level of bovine serum albumin in milk, peaked at 4 h and then gradually decreased . All parameters remained at basal levels in control quarters throughout the study . In vitro experiments indicated that neutrophils released sCD14 in response to LPS in a dose-dependent manner . The results indicate that the concentration of sCD14 was significantly increased in milk after LPS challenge . The increase was not likely due to serum leakage . Instead, infiltrated neutrophils might be the main source of increased sCD14 in milk during inflammation. J Clin Microbiol, 2003 Aug, 41(8), 3942 - 4 Clinical impact of a PCR assay for identification of Staphylococcus aureus and determination of methicillin resistance directly from blood cultures; Hallin M et al.; We evaluated the clinical usefulness of a PCR assay that discriminates Staphylococcus aureus from coagulase-negative staphylococci and detects methicillin resistance on blood cultures by measuring the adaptation of antimicrobial therapy based on the PCR results . Only 7 of 28 patients (25%) benefited from a modification of antibiotic therapy based on the PCR results, since empirical therapy was appropriate in a majority of cases. J Clin Microbiol, 2003 Aug, 41(8), 3890 - 2 Detection of methicillin-resistant Staphylococcus aureus (MRSA) in blood with the EVIGENE MRSA detection kit; Levi K et al.; A total of 200 blood cultures containing putative staphylococci were analyzed by a commercial gene probe hybridization assay (EVIGENE; Statens Serum Institut, Copenhagen, Denmark), and 18 were identified as methicillin-resistant Staphylococcus aureus (MRSA) positive . Of these, 17 were positive by PCR and 16 were positive by culture . Detailed analysis of the discrepant results showed that the EVIGENE kit allowed specific identification of MRSA in blood cultures without any of the drawbacks associated with PCR. J Clin Microbiol, 2003 Aug, 41(8), 3609 - 14 Coagulase-negative staphylococci: comparison of phenotypic and genotypic oxacillin susceptibility tests and evaluation of the agar screening test by using different concentrations of oxacillin; Ferreira RB et al.; This study evaluated the oxacillin susceptibilities of 152 coagulase-negative staphylococcal (CoNS) strains of 12 species by disk diffusion; agar dilution; E-test; the slide latex agglutination test (Slidex MRSA Detection test; bioMerieux S/A, Paris, France); the agar screening test with 1, 2, 4, or 6 microg of oxacillin per ml and incubation for 24 or 48 h; and detection of the mecA gene by PCR . The results revealed that the agar screening test with 4 micro g of oxacillin per ml and incubation for 48 h was superior to any single phenotype-based susceptibility assay, presenting a sensitivity and a specificity of 100% each . For the different methods evaluated, the sensitivities and specificities were as follows: for disk diffusion, 94.2 and 91.8%, respectively; for the agar dilution test 100 and 73.5%, respectively; for E-test, 100 and 71.4%, respectively; and for the slide latex agglutination test, 97.1 and 98%, respectively . A good correlation was observed between oxacillin susceptibility testing results and PCR results for Staphylococcus epidermidis, S . haemolyticus, S . hominis subsp . hominis, and all mecA-positive strains . However, at least 60% of the mecA-negative isolates of the species S . saprophyticus, S . cohnii subsp . urealyticum, S . lugdunensis, and S . sciuri were erroneously classified as oxacillin resistant by the agar dilution test . Conversely, the slide latex agglutination test presented a high sensitivity (97.1%) and a high specificity (98%) for all CoNS species . Our results demonstrated the accuracy of the agar screening test with 4 micro g of oxacillin per ml and incubation for 48 h and the slide latex agglutination test for the appropriate detection of the oxacillin susceptibilities of CoNS isolates . Both assays are technically simple and can be easier to perform in routine laboratories than PCR. J Clin Microbiol, 2003 Aug, 41(8), 3499 - 502 Evaluation of a novel medium for screening specimens from hospitalized patients to detect methicillin-resistant Staphylococcus aureus; Blanc DS et al.; A novel medium, Oxacillin Resistant Screening Agar (ORSA) medium, was evaluated for the screening of specimens for methicillin-resistant Staphylococcus aureus (MRSA) in the hospital setting . Screening swabs (swabs of the nose, throat, perineum, and infected sites) were inoculated onto the new ORSA medium and into an enrichment broth (Muller-Hinton broth supplemented with NaCl and oxacillin) . After 24 h of incubation, the enrichment broth was subcultured onto one ORSA plate and one lipovitellin Chapman salt agar plate . The sensitivities for the detection of MRSA were calculated for each medium alone and for the media in combination . A low sensitivity (74%) was obtained when ORSA medium was used alone as a primary culture, whereas the sensitivity was 88% when a single selective enrichment broth was used . Among the 414 blue colonies observed on ORSA plates, only 47% were found to be MRSA, 40% were coagulase-negative staphylococci, 7% were Enterococcus species, and 2% were methicillin-sensitive S . aureus . The optimal incubation time for the ORSA plates was evaluated . On primary culture, 38% of the blue MRSA colonies were visible only after 48 h of incubation (no blue colonies were not seen after 24 h of incubation), whereas 94% of the colonies were already visible at 24 h when ORSA plates were used for subcultures . In conclusion, the advantage of the novel ORSA medium is the ease of recognition of mannitol-fermenting bacteria, but further identification tests are needed to confirm the identification of S . aureus . An enrichment broth is still needed to ensure a good sensitivity for the recovery of MRSA, and an incubation time of 48 h is required for primary culture on ORSA medium. Biomaterials, 2003 Aug, 24(18), 3013 - 9 Staphylococcus epidermidis-fibronectin binding and its inhibition by heparin; Arciola CR et al.; Staphylococcus epidermidis is able to adhere onto biomaterials and to cause implant infections . Recently, host matrix proteins, which in vivo cover the implants, have been indicated as substrates for adhesion by specific bacterial adhesins . Here, the binding of S . epidermidis to fibronectin, a main protein of the extracellular matrix, and the effect of heparin on this interaction were studied by dynamic force spectroscopy (DFS) . Novelties are that S . epidermidis strains analysed by DFS were clinical isolates from prosthesis-associated infections, genotyped and phenotyped for their adhesion properties to fibronectin and examined as living cells . Thus, fibronectin-binding staphylococci adhered to the fibronectin-coated substratum and formed a continuous layer assuring their contact with the fibronectin-coated cantilever tip during the approach-retraction cycles of the DFS measurements . Results show that only a single molecular binding site of fibronectin is involved in the interaction with S . epidermidis, that it takes place at the domain near the C-terminus and that it is specifically inhibited by heparin. Mol Microbiol, 2003 Aug, 49(4), 919 - 27 Biphasic intracellular expression of Staphylococcus aureus virulence factors and evidence for Agr-mediated diffusion sensing; Shompole S et al.; Staphylococcus aureus invades a variety of mammalian cells and escapes from the endosome to multiply in the cytoplasm . We had previously hypothesized that the molecular events leading to escape of S . aureus from the endosome involved the Agr virulence factor regulatory system . In this report we demonstrate that temporal changes in intracellular activation of the Agr regulon correlates with expression of membrane active toxins . Also, the initial expression of Agr by even small numbers of staphylococci resulted in the permeabilization of the endosomal membrane and the eventual escape of bacteria into the cytoplasm by 3 h post invasion . After Agr downregulation, a second peak of expression coincided with increased permeability of the host cell membrane . In contrast to the parental strain, an Agr-mutant was unable to escape into the cytoplasm and was observed in intact endosomes as late as 5 h post invasion . These data provide evidence that staphylococcal virulence factor production during invasion of host cells is mediated by an Agr-dependent process that is most accurately described in the context of diffusion sensing. Lik Sprava, 2003 Apr-Jun, (3-4), 90 - 2 {Species affiliation of staphylococci isolated in purulent infections.}; Nuruzova ZA et al.; The microflora was studied of the clinical material taken from those patients presenting with different pyo-septic diseases . It has been ascertained that a key role in the etiology of the above diseases must be referred to gramme-positive micro-organisms--to coagulase-negative staphylococci among their number . Of these, S . haemolyticus and S . epidermidis were found with high frequency. Lik Sprava, 2003 Apr-Jun, (3-4), 72 - 4 {Treatment of patients with acute pneumonia}; Toropchin VI et al.; Efficiency was studied of treatment of acute pneumonia (AP) in 148 patients with a focal croupous form . In the grave course of AP, antibacterial therapy with benzilpenicillin combined with sulphanilamide agents biseptol and nitrofuran was tried, the correcting therapy having been instituted on obtaining findings from assays of the sputum for sensitivity of the microflora to antibiotics . In those cases with Friendlander's bacilli recoverable, methicillin, oxacillin, macrolids, cephalosporins and some other agents were antibiotics of choice . Administered in the atypical course that was related more frequently to mycoplasms or chlamidia, was erythromycin phosphate i.v., lincomycin . In Staphylococcus-associated pneumonia, there have been used antibiotics resistant to penicillinases and endowed with an inhibitory activity toward staphylococci--oxacillin, methicillin, cephalosporins, chlorophenicols or chlorophphyllipt i.v . combined with an antistaphylacoccal plasma, antistaphylococcal gammaglobulin or hyperimmune plasma . The following drugs were prescribed to ensure bronchial patency and liquefaction of the sputum--euphyllin, mucalthin, bromhexin, lasolvan that are known to stimulate production of sulfoctant . The therapeutic complex comprized fibs, aloe, apilac, prodigiosan, solcoseryl, methyluracil known to stimulate bodily reactivity and resolution of inflammatory infiltration, nicotinic acid, heparin that have been shown to improve micorcirculation, tocopherol, unithiol known to regulate lipid oxidation . The data secured suggest to us a sufficient efficacy of the above-outlined therapy of AP. Zh Mikrobiol Epidemiol Immunobiol, 2003 May-Jun, (3), 99 - 109 {Molecular epidemiology of infections caused by methicillin-resistant staphylococci}; Shaginian IA et al.; The review deals with the periodicity of the spread of methicillin-resistant S . aureus (MRSA) strains during the last 40 years, the mechanism of their resistance to methicillin and other beta-lactamic antibiotics, the genetic control of methicillin resistance, the genome organization of mec DNA and its possible cause, as well as the organization of epidemiological surveillance on MSRA in hospitals . The problem of changes in the epidemiology of staphylococcal infections due to the appearance of MRSA in the absence of contacts with carriers, treatment with antibiotics or stay in a hospital is discussed . The concern of public health authorities in connection with the emergence of MRSA strains, moderately resistant or resistant to vancomycin, is also discussed . The most promising programs of the MRSA study, as well as the optimum programs introduced in economically developed counties for the control of hospital infections caused by MRSA, are considered. Circulation, 2003 Aug 12, 108(6), 767 - 71 Epub 2003 Jul 28. Prophylactic efficacy of topical temporin A and RNAIII-inhibiting peptide in a subcutaneous rat Pouch model of graft infection attributable to staphylococci with intermediate resistance to glycopeptides; Cirioni O et al.; BACKGROUND: Bacteria that adhere to implanted medical devices play an important role in industry and in modern medicine . Staphylococci are among the most common pathogens that cause biomaterial infections . Vascular prosthetic graft infection is one of the most feared complications that the vascular surgeon treats, frequently resulting in prolonged hospitalization, organ failure, amputation, and death . A rat model was used to investigate the topical efficacies of temporin A and the quorum-sensing inhibitor RNAIII-inhibiting protein (RIP) as prophylactic agents of vascular prosthetic graft infections caused by Staphylococcus aureus and Staphylococcus epidermidis with intermediate resistance to glycopeptides . METHODS AND RESULTS: Graft infections were established in the back subcutaneous tissue of adult male Wistar rats by implantation of Dacron prostheses 1 cm2 followed by topical inoculation with 2x10(7) colony-forming units of bacterial strains . The study included, for each staphylococcal strain, a control group (no graft contamination), a contaminated group that did not receive antibiotic prophylaxis, and 6 contaminated groups that received grafts soaked with temporin A, RIP, rifampin, temporin A plus RIP, RIP plus rifampin, or temporin A plus RIP . The infection was evaluated by quantitative agar culture . When tested alone, temporin A and RIP showed comparable efficacies, and their efficacies were significantly higher than that of rifampin against both strains . All combinations showed efficacies significantly higher than that of each single compound . The combinations of temporin A and RIP exerted the strongest antistaphylococcal efficacies, eliminating infection by 100% . CONCLUSIONS: The results of the present study make these molecules potentially useful for antimicrobial chemoprophylaxis in vascular surgery. Aviakosm Ekolog Med, 2003, 37(3), 28 - 31 {Effects of high-pressure argon-containing gaseous compositions on operators' rhinopharynx and external ear microflora and its reactivity to antibiotics}; Solov'eva ZO et al.; Earlier investigations revealed an important microbiological feature of the human in hyperbaric environment--linear progressive unbalance in the opportunistic and commensal ratio within the main biotopes . Change in the gaseous environment may influence the activity of infectious agents . Purpose of our investigation was to study effects of argon-containing gaseous compositions on the rhinopharynx and external ear microflora in operators, and its reactivity to antibiotics . Attempted was quantification of the operators' aerobic microflora exposed for 6, 7 and 18 days to argon-containing gaseous compositions (normoxic and hypoxic) under an elevated pressure (0.15-0.5 MPa) . Evaluated was also susceptibility of the human microflora to antibiotics of the basic chemical classes under these conditions . The argon-containing gaseous compositions in the above pressure range did not have any specific effects on the ratio of aerobic gram-negative bacilli and staphylococci; neither they created selective advantages to any of the groups of microorganisms under study . It was found additionally that long-duration exposure in the argon-containing gaseous environment (6, 7 and 18 days) gave rise to cultures with the signs of hospitalism (e.g., high index of multiple drug resistance) . Potentiality of infection development in dwelling modules with argon-containing gaseous environment and proliferation of polyresistant strains point to the necessity to design a respective preventive system. Environ Microbiol, 2003 Aug, 5(8), 711 - 6 Impact of antibiotics on conjugational resistance gene transfer in Staphylococcus aureus in sewage; Ohlsen K et al.; The growing rate of microbial pathogens becoming resistant to standard antibiotics is an important threat to public health . In order to assess the role of antibiotics in the environment on the spread of resistance factors, the impact of subinhibitory concentrations of antibiotics in sewage on gene transfer was investigated using conjugative gentamicin resistance (aacA-aphD) plasmids of Staphylococcus aureus . Furthermore, the concentration of antibiotics in hospital sewage was measured by high-performance liquid chromatography (HPLC)-electrospray tandem mass spectrometry . Several antibiotics were found to be present in sewage, e.g . ciprofloxacin up to 0.051 mgl(-1) and erythromycin up to 0.027 mgl(-1) . Resistance plasmid transfer occurred both on solidified (dewatered) sewage and in liquid sewage in a bioreactor with a frequency of 1.1x10(-5)-5.0x10(-8) . However, low-level concentrations of antibiotics measured in sewage are below concentrations that can increase plasmid transfer frequencies of gentamicin resistance plasmids of staphylococci. Transfusion, 2003 Aug, 43(8), 1047 - 52 Estimation of bacterial risk in extending the shelf life of PLT concentrates from 5 to 7 days; Lee CK et al.; BACKGROUND: The use of bacterial culture to prevent bacterial contamination of blood components has renewed interest for extending the shelf life of PLT concentrates to 7 days after collection . STUDY DESIGN AND METHODS: This study was therefore conducted to determine the residual risk of bacterial contamination in PLT concentrates at the end of 5 and 7 days after collection in a center where all PLT concentrates are routinely screened by taking samples on Day 2 for culture . PLT units with no growth after 48 hours were sampled a second time on Day 5 or Day 7 after collection, followed by inoculation into aerobic culture bottles . The inoculated bottles were then monitored for up to 7 days at 35 degrees C in an automatic monitoring and detection system . RESULTS: During a 16-month study period, a total of 6020 PLT concentrates were tested 5 days (Group A, n=3010) and 7 days (Group B, n=3010) after collection . Four units in each group (0.133%) were found to be contaminated . In 6 units, bacteria were seen on direct Gram stain . In addition, 5 of the associated RBC units grew the same organisms on culture . The organisms include three coagulase-negative staphylococci and five Propionibacterium acnes . The positive rate of routine short-term bacterial culture was 0.035 percent during the same study period . CONCLUSION: Despite routine short-term bacterial culture, a significant risk of bacterial contamination remains at 5 and 7 days after collection . For now, the shelf life of PLT concentrates should remain 5 days. Curr Infect Dis Rep, 2003 Aug, 5(4), 293 - 299 Infective Endocarditis in Patients with Kidney Failure: Chronic Dialysis and Kidney Transplant; Ireland JH et al.; Physicians who treat patients with infective endocarditis (IE) are encountering a growing number of dialysis and kidney transplant patients . Both groups have 30 to 100 times higher risk of IE, with 1-year mortalities of 40% to 60% . The predominant organisms causing IE are gram positive, with 60% to 80% of cases due to Staphylococcus aureus, and another 10% to 20% of cases due to coagulase-negative staphylococci . Renal transplant patients may develop fungal IE, but this risk is primarily in the first 3 months after transplant . In addition to blood cultures, transesophageal echocardiogram is the most useful diagnostic examination for IE in these patients . Initial antibiotic therapy, pending final culture and antibiotic susceptibility results, should provide coverage against the most common organisms and allow for the potential of either methicillin or vancomycin-resistant species . Removal of infected hemodialysis access devices and at least 4 to 6 weeks of intravenous antibiotics are recommended . Antibiotic prophylaxis against IE has been recommended for all dialysis and renal transplant patients, but this strategy is controversial and unproven. Br J Biomed Sci, 2003, 60(2), 71 - 4 Screening for methicillin resistance in Staphylococcus aureus and coagulase-negative staphylococci: an evaluation of three selective media and Mastalex-MRSA latex agglutination; Bowers KM et al.; Laboratory confirmation of MRSA is important for the implementation of infection control; conventional screening culture methods take up to five days for confirmation . The purpose of this study is to ascertain the efficiency of three selective media for growth of methicillin-resistant Staphylococcus aureus (MRSA) before and after enrichment in salt broth, and to evaluate the Mastalex-MRSA latex agglutination kit for detection of methicillin resistance . Screening swabs were collected from 63 patients, yielding 125 S . aureus isolates and 40 coagulase-negative staphylococcus (CNS) isolates . Selective media used were mannitol salt agar (MSA), Baird-Parker agar with ciprofloxacin (BPC) and bromocresol purple (BCPA) . Polymerase chain reaction (PCR) for mecA gene detection was used as the reference standard for evaluation of the Mastalex-MRSA assay, which was also evaluated on colonies of S . aureus from the selective media . No significant difference was found in efficiency of MRSA isolation among the selective media . Pre-enrichment in the salt broth did not enhance isolation of MRSA . Methicillin-sensitive S . aureus and CNS were significantly inhibited in all selective media (P<0.05) . Only BPC significantly selected out methicillin-resistant CNS (P<0.01) . Mastalex-MRSA was 97% specific and sensitive for the detection of MRSA . It was 65% sensitive and 100% specific in detecting methicillin resistance in CNS . In conclusion, all selective media performed equally well (MRSA isolation rate of approximately 80%) . Mastalex-MRSA provided rapid and reliable detection of MRSA from selective media, reducing the time required for confirmation of this organism. Nahrung, 2003 Jun, 47(3), 166 - 70 Differentiation of toxigenic Staphylococcus aureus in staphylococcal isolates from prepared and frozen foods by combined arbitrarily primed polymerase chain reaction and DNA probe; Cordoba MG et al.; In prepared and frozen flamenquin and hake fish fingers Staphylococcus aureus as sanitary hazards have been detected . In the present work, a combined method that includes an arbitrarily primed PCR (AP-PCR) and a mixed DNA probe hybridisation designed for the enterotoxigenic genes sea, seb, sec, and sed will be assayed to differentiate enterotoxigenic S . aureus from other staphylococcal species isolated during the processing of prepared and frozen foods . From the protocols tested for the AP-PCR, the highest number of amplification bands showing the best resolution was achieved at 30 degrees C annealing and 35 degrees C extension temperatures . Several staphylococci identified by a biochemical test as S . aureus showed in the AP-PCR analysis different banding patterns to the references S . aureus . The isolates, were investigated by slot blot hybridisation for genes encoding A, B, C, and D staphylococcal enterotoxins to determine their enterotoxigenic potential . Several isolates characterised by the AP-PCR analysis as S . aureus hybridised with the DNA probe mixture . The combined AP-PCR and DNA probe hybridisation assayed was able to differentiate toxigenic S . aureus from other staphylococcal species from prepared and frozen foods . This method could be considered as microbial quality assurance in these products. J Antimicrob Chemother, 2003 Aug, 52(2), 258 - 63 Epub 2003 Jul 15. Fitness of antibiotic resistant Staphylococcus epidermidis assessed by competition on the skin of human volunteers; Gustafsson I et al.; BACKGROUND: Antibiotic resistance typically confers a biological fitness cost on bacteria that can be manifested as a decreased growth rate in culture media and experimental animals . However, there are limited experimental data on the relative fitness of resistant and susceptible bacteria during growth in their natural environment . OBJECTIVE: We have developed a human competition model to investigate the relative fitness of antibiotic-resistant and -susceptible bacteria . MATERIALS AND METHODS: A non-epidemic Staphylococcus epidermidis strain was isolated from skin, and a rifampicin-resistant (RifR) clone was selected . The RifR marker was used to distinguish the inoculated strains from the resident population of coagulase-negative staphylococci . The RifR strains were further selected for resistance to ciprofloxacin (CipR) and fusidic acid (FusR) . A 1:1 mix of susceptible and resistant bacteria was applied on the forearms of 12 volunteers . Competition was monitored by sampling bacteria from skin and determining their relative numbers . RESULTS: Resistance to ciprofloxacin due to parC mutations did not decrease the growth rate in vitro, and the CipR/CipS ratio was close to 1 during day 1 and 3 in the in vivo competition experiments . In contrast, fusidic acid resistance due to fusA mutations resulted in a decrease in the growth rate in vitro and a considerable loss of fitness in the competition . The FusR/FusS ratio diminished from 1.3 to 0.023 in 3 days . CONCLUSIONS: These data show that human volunteers can be used as a simple and relevant model to study the biological cost of resistance. East Afr Med J, 2000 Jul, 77(7), 386 - 90 Ventriculoperitoneal shunt surgery and shunt infections in children with non-tumour hydrocephalus at the Kenyatta National Hospital, Nairobi; Mwang'ombe NJ et al.; OBJECTIVE: To study infections complicating ventriculoperitoneal (VP) shunt surgery in children with non-tumour hydrocephalus at the Kenyatta National Hospital, Nairobi . DESIGN: A retrospective survey . SETTING: Kenyatta National Hospital, Nairobi between January 1982 and December 1991 . SUBJECTS: Three hundred and forty five patients who underwent V-P shunt placement for non-tumour hydrocephalus . RESULTS: Three hundred and forty five patients underwent V-P shunt placement for non-tumour hydrocephalus . There were 107 infection episodes involving 85 patients . The ages of these patients ranged from three months to 12 years . Most of the patients had congenital hydrocephalus . The infection rate was high (24.6%) although comparable to infection rates reported for clean surgery in the hospital . Fever, septic wounds and features of shunt malfunction were the main presenting features . Bacteriological studies confirmed Staphylococcus aureus and coagulase negative staphylococci as the two most commonly isolated micro-organisms . CONCLUSION: This study emphasises need to reduce infection rate in ventriculoperitoneal shunt surgery at the Kenyatta National Hospital . Definitive surgical treatment for hydrocephalus was in most cases delayed and this problem was also observed during revision of infected shunts . Late presentation was often due to ignorance and the fact that many patients went for traditional forms of treatment first before going to hospital. Microbes Infect, 2003 Jul, 5(9), 775 - 80 The role of Staphylococcus aureus sortase A and sortase B in murine arthritis; Jonsson IM et al.; Gram-positive pathogenic bacteria display proteins on their surface that play important roles during infection . In Staphylococcus aureus, these surface proteins are anchored to the cell wall by two sortase enzymes, SrtA and SrtB, that recognize specific surface protein sorting signals . The role of sortase enzymes in bacterial virulence was examined using a murine septic arthritis model . Intravenous inoculation with any of the Delta(srtA), Delta(srtB) or Delta(srtAB) mutants resulted in significantly increased survival and significantly lower weight loss compared with the parental strain . Mice inoculated with the Delta(srtA) mutant did not express severe arthritis, while arthritis in mice inoculated with the Delta(srtB) mutant was not different from that seen in mice that were infected with the wild-type parent strain . Furthermore, persistence of staphylococci in kidneys and joints following intravenous inoculation of mice was more pronounced for wild-type and Delta(srtB) mutant strains than for Delta(srtA) or Delta(srtAB) variants . Together these results indicate that sortase B (srtB) plays a contributing role during the pathogenesis of staphylococcal infections, whereas sortase A (srtA) is an essential virulence factor for the establishment of septic arthritis. Echocardiography, 2003 Apr, 20(3), 289 - 90 Role of transesophageal echocardiography in detecting implantable cardioverter defibrillator lead infection; Wasson S et al.; Implantable cardioverter defibrillator (ICD) lead infection is a rare condition with reported incidence of 0.2% to 16% . It usually presents with persistent bacteremia or fever of unknown origin and requires high clinical suspicion for diagnosis . Whenever ICD lead infection is suspected, transesophageal echocardiography is the diagnostic technique of choice for detection and characterization of the lesions . Lead infections are extremely difficult to manage conservatively and surgical removal of the entire defibrillator system is recommended along with antimicrobial therapy . We describe a case of recurrent staphylococci bacteremia due to an ICD lead infection in a patient with arrhythmogenic right ventricular dysplasia. Eur J Clin Microbiol Infect Dis, 2003 Aug, 22(8), 489 - 91 Epub 2003 Jul 04. Staphylococcus lugdunensis endocarditis--the hidden peril of coagulase-negative staphylococcus in blood cultures; Seenivasan MH et al.; Reported here is a successfully treated case of native mitral valve endocarditis caused by Staphylococcus lugdunensis and a review of 47 similar cases reported in the English literature . In the literature review, perineal skin flora appeared to be the source of the organism in patients with endocarditis . Staphylococcus lugdunensis is generally susceptible in vitro to beta-lactam agents . If speciation is not performed, these bacteria might be mistaken for Staphylococcus epidermidis, a relatively avirulent bacterium that is a common contaminant of cultures . Prompt speciation can lead to earlier recognition of endocarditis and possibly enable earlier surgical intervention with improved outcome for this high-mortality infection . Multiple positive blood cultures yielding coagulase-negative staphylococci should be identified to the species level; endocarditis or another intravascular source of infection should be sought. Infez Med, 1997, 5(4), 214 - 29 {Antimicrobial prophylaxis in clean surgery}; de Lalla F; Antibiotic prophylaxis in clean surgery is generally not required except for prosthetic cardiac, vascular and orthopedic surgery . In these cases, the cephalosporins mostly active against staphylococci such as cefazolin, cefuroxime and cefamandole or antistaphylococcal penicillins, such as oxacillin, associated or not with aminoglycosides, are recommended . The progressive increase of methicillin-resistant staphylococci has led to use for the above mentioned surgeries also glycopeptides, especially in those wards where methicillin resistance is particularly high . Nevertheless, not enough many prospective-controlled studies have been carried on to evaluate whether glycopeptides reduce the risk of post-surgical infections, more than other conventional antibiotic regimens . Further, the possible risk of selecting glycopeptide-resistant enterococci suggests that prophylactic use of glycopeptides should be limited and a minimal number of doses should be administered, favouring whenever possible, the ultra short term prophylaxis (single dose). Ann Hematol, 2003 Aug, 82(8), 526 - 8 Epub 2003 Jul 03. Henoch-Schönlein IgA glomerulonephritis complicating myeloma kidneys: case report; Arrizabalaga P et al.; Myeloma kidney is the principal pathological substrate of rapidly progressive renal failure in multiple myeloma . We report the unusual case of a 72-year-old male diagnosed with kappa Bence Jones myeloma with renal failure which needed dialysis . After treatment with vincristine, doxorubicin (Adriamycin), dexamethasone (VAD), and plasmapheresis, the renal function was recovered until serum creatinine level was <2 mg/dl . Six months later, the pathological counterpart of rapidly progressive renal failure was crescentic IgA proliferative glomerulonephritis as a manifestation of Henoch-Schonlein syndrome associated with sepsis caused by coagulase-negative staphylococci . This case suggests that mesangial IgA deposition should be considered within the spectrum of consequent glomerular lesion-associated chemotherapy occurring in multiple myeloma. Mikrobiyol Bul, 2002 Jul-Oct, 36(3-4), 253 - 7 {Comparison of methods used to detect methicillin resistance in staphylococci}; Kuzucu C et al.; In this study, the methicillin resistance of 112 Staphylococcus aureus and 93 coagulase negative Staphylococcus (CNS) strains, which were initially found methicillin resistant by routine disk diffusion method in our laboratory, have been searched by microdilution and oxacillin agar screen test and the results were compared with the results obtained by disk diffusion method . The presence of mecA gene was investigated by polymerase chain reaction in case of discordant results . All S . aureus strains (100%) and 69.9% (65/93) of CNS strains were found resistant to methicillin by three of the methods . Of CNS isolates, 28 strains which were found methicillin resistant by disk diffusion method, were found methicillin susceptible by oxacillin agar screen method, and 27 of these were detected as mecA positive . Our results indicated that, the three methods tested were reliable for the detection of methicillin resistance in S . aureus strains, but oxacillin agar screen revealed to be unsatisfactory for the detection of methicillin resistance in CNS. Pediatr Int, 2003 Jun, 45(3), 301 - 6 Efficacy and safety of arbekacin for staphylococcal infection in the NICU; Suzuki K; BACKGROUND: This is a retrospective study on the efficacy and safety of arbekacin (ABK), an aminoglycoside antibiotic, for acquired staphylococcal infection in the neonatal intensive care nursery . PATIENTS AND METHODS: Subjects were 29 infants treated with ABK in a tertiary care neonatal center . They were 23-39 (median 28) weeks' gestation, 530-3334 (median 930) grams at birth, and 3-157 (median 17) days of age . Diagnosis of staphylococcal infection was made by clinical signs and laboratory findings . Sensitivity of the isolated organisms to ABK was tested by the microliquid dilution method . Serum ABK level was monitored to achieve the therapeutic range during the treatment . Effectiveness was defined by improving clinical signs and laboratory findings within 3 days . Effectiveness was studied in relation to type of infection and other antibiotics administered . Auditory brainstem response and serum creatinine changes were studied for ototoxicity and nephrotoxicity assessment, respectively . RESULTS: Twenty-seven (93.1%) cases of infection were attributed to methicillin-resistant Staphylococcus aureus (MRSA) and two (6.9%) were attributed to coagulase-negative staphylococci (CNS) . The rate of in vitro sensitivity to ABK was 85.2% for MRSA and 100.0% for CNS . The overall clinical effeciveness rate was 79.3% (23/29) with no difference associated with types of infection . Combination of ABK with sulbactam/ampicillin showed greater effectiveness (100.0%) than with other antibiotics (64.3%) (P < 0.05) . There was no abnormal auditory brainstem response or serum creatinine change associated with ABK treatment . CONCLUSION: ABK is an effective and safe antibiotic for the treatment of acquired staphylococcal infection in the neonatal intensive care nursery. Pediatr Infect Dis J, 2003 Jun, 22(6), 572 - 3 Primary osteomyelitis and suppurative arthritis caused by coagulase-negative staphylococci in a preterm neonate; Eggink BH et al.; Coagulase-negative staphylococci are a major cause of nosocomial infections in neonatal intensive care unit patients . These infections are usually related to the presence of intravascular devices . An 1175-g preterm neonate developed primary osteomyelitis and septic arthritis by coagulase-negative staphylococci in the absence of any indwelling central catheters. Prescrire Int, 2003 Jun, 12(65), 108 - 9 Acute otitis media in adults: many unknowns. {Antimicrobial activity of fosfomycin against beta-lactamase-producing methicillin-sensitive Staphylococcus aureus and methicillin-sensitive coagulase-negative staphylococci} Hara T, Araake M, Tsuruoka T, Watabe H. Pharmaceutical Research Center, Meiji Seika Kaisha, Ltd., 760 Morooka-cho, Kohoku-ku, Yokohama, Kanagawa, 222-8567, JapanAntimicrobial activity of fosfomycin (FOM), cefazolin (CEZ), cefmetazole (CMZ), cefotiam (CTM) and piperacillin (PIPC) against clinical isolates of methicillin-sensitive Staphylococcus aureus (MSSA) (beta-lactamase-producing or non-producing) and methicillin-sensitive coagulase-negative Staphylococci (MSCNS) (beta-lactamase-producing or non-producing) were determined to make clear the differences in antimicrobial activity of FOM and beta-lactam antibiotics . The antimicrobial activity of PIPC against beta-lactamase-producing strains of MSSA was lower than that against non-producing ones, judging from the distribution patterns of susceptibility of the strains to PIPC . There were no differences in the antimicrobial activity of FOM, CEZ and CMZ for the producing and non-producing strains . The activity of FOM against MSCNS was comparable to that against MSSA, although those of CEZ, CMZ, CTM and PIPC were decreased . FOM, CEZ, CMZ and CTM showed bactericidal activity against TH4278 (MIC {microgram/ml}: FOM, 1; CEZ, 0.5; CMZ, 1; CTM, 0.5; PIPC, 1) of beta-lactamase-producing MSSA at 1 microgram/ml for 6 h, but PIPC did not at the same condition . FOM and CMZ at MIC suppressed regrowth of the strain, but CEZ, CTM and PIPC did not . In conclusion, FOM, which is not affected by beta-lactamase, demonstrated strong bactericidal activity at low concentration against the beta-lactam-resistant strains due to beta-lactamase production. Arch Pathol Lab Med, 2003 Jul, 127(7), 845 - 9 Development and validation of a molecular beacon probe-based real-time polymerase chain reaction assay for rapid detection of methicillin resistance in Staphylococcus aureus; Elsayed S et al.; CONTEXT: A rapid, real-time, duplex, fluorescent molecular beacon probe-based polymerase chain reaction (PCR) assay was recently