Resistant biofilms formed by Staphylococcus aureus on medical devices pose a constant medical threat. A promising alternative to tackle this problem is photodynamic inactivation (PDI). This study focuses on a polyurethane (PU) material with an antimicrobial surface consisting of a composite based on silicate, polycation, and erythrosine B (EryB). The composite was characterized using X-ray diffraction and spectroscopy methods. Anti-biofilm effectiveness was determined after PDI by calculation of CFU mL-1. The liquid PU precursors penetrated a thin silicate film resulting in effective binding of the PU/silicate composite and the PU bulk phases. The incorporation of EryB into the composite matrix did not significantly alter the spectral properties or photoactivity of the dye. A green LED lamp and laser were used for PDI, while irradiation was performed for different periods. Preliminary experiments with EryB solutions on planktonic cells and biofilms optimized the conditions for PDI on the nanocomposite materials. Significant eradication of S. aureus biofilm on the composite surface was achieved by irradiation with an LED lamp and laser for 1.5 h and 10 min, respectively, resulting in a 10,000-fold reduction in biofilm growth. These results demonstrate potential for the development of antimicrobial polymer surfaces for modification of medical materials and devices.

Few methicillin-resistant Staphylococcus aureus (MRSA) from the early years of its global emergence have been sequenced. Knowledge about evolutionary factors promoting the success of specific MRSA multi-locus sequence types (MLSTs) remains scarce. We aimed to characterize a legacy MRSA collection isolated from 1965 to 1987 and compare it against publicly available international and local genomes.

We accessed 451 historic (1965-1987) MRSA isolates stored in the Culture Collection of Switzerland, mostly collected from the Zurich region. We determined phenotypic antimicrobial resistance (AMR) and performed whole genome sequencing (WGS) using Illumina short-read sequencing on all isolates and long-read sequencing on a selection with Oxford Nanopore Technology. For context, we included 103 publicly available international assemblies from 1960 to 1992 and sequenced 1207 modern Swiss MRSA isolates from 2007 to 2022. We analyzed the core genome (cg)MLST and predicted SCCmec cassette types, AMR, and virulence genes.

Among the 451 historic Swiss MRSA isolates, we found 17 sequence types (STs) of which 11 have been previously described. Two STs were novel combinations of known loci and six isolates carried previously unsubmitted MLST alleles, representing five new STs (ST7843, ST7844, ST7837, ST7839, and ST7842). Most isolates (83% 376/451) represented ST247-MRSA-I isolated in the 1960s, followed by ST7844 (6% 25/451), a novel single locus variant (SLV) of ST239. Analysis by cgMLST indicated that isolates belonging to ST7844-MRSA-III cluster within the diversity of ST239-MRSA-III. Early MRSA were predominantly from clonal complex (CC)8. From 1980 to the end of the twentieth century, we observed that CC22 and CC5 as well as CC8 were present, both locally and internationally.

The combined analysis of 1761 historic and contemporary MRSA isolates across more than 50 years uncovered novel STs and allowed us a glimpse into the lineage flux between Swiss-German and international MRSA across time.

The horizontal transmission of methicillin resistance to Staphylococcus aureus (MRSA) in hospital and community settings, and growing prevalence of these strains, presents a significant clinical challenge to the management of serious infections worldwide. While infection control initiatives have stemmed the rising prevalence, MRSA remains a significant pathogen. More recently, evidence that MRSA is becoming resistant to glycopeptides and newer therapies raises concern about the use of these therapies in clinical practice. Vancomycin resistance has become evident in select clinical settings through rising MICs, growing awareness of heteroresistance, and emergence of intermediate-resistant and fully resistant strains. While resistance to linezolid and daptomycin remains low overall, point mutations leading to resistance have been described for linezolid, and horizontal transmission of cfr-mediated resistance to linezolid has been reported in clinical isolates. These resistance trends for newer therapies highlight the ongoing need for new and more potent antimicrobial therapies.

The absence of rapid tests evaluating antibiotic susceptibility results in the empirical prescription of antibiotics. This can lead to treatment failures due to escalating antibiotic resistance, and also furthers the emergence of drug-resistant bacteria. This study reports a rapid optical method to detect β-lactamase and thereby assess activity of β-lactam antibiotics, which could provide an approach for targeted prescription of antibiotics. The methodology is centred on a fluorescence quenching based probe (β-LEAF--β-Lactamase Enzyme Activated Fluorophore) that mimics the structure of β-lactam antibiotics.

The β-LEAF assay was performed for rapid determination of β-lactamase production and activity of β-lactam antibiotic (cefazolin) on a panel of Staphylococcus aureus ATCC strains and clinical isolates. Four of the clinical isolates were determined to be lactamase producers, with the capacity to inactivate cefazolin, out of the twenty-five isolates tested. These results were compared against gold standard methods, nitrocefin disk test for β-lactamase detection and disk diffusion for antibiotic susceptibility, showing results to be largely consistent. Furthermore, in the sub-set of β-lactamase producers, it was demonstrated and validated that multiple antibiotics (cefazolin, cefoxitin, cefepime) could be assessed simultaneously to predict the antibiotic that would be most active for a given bacterial isolate.

The study establishes the rapid β-LEAF assay for β-lactamase detection and prediction of antibiotic activity using S. aureus clinical isolates. Although the focus in the current study is β-lactamase-based resistance, the overall approach represents a broad diagnostic platform. In the long-term, these studies form the basis for the development of assays utilizing a broader variety of targets, pathogens and drugs.

Staphylococcus aureus is a gram-positive bacterium that colonises the skin and is present in the anterior nares in about 25-30% of healthy people. Dependent on its intrinsic virulence or the ability of the host to contain its opportunistic behaviour, S aureus can cause a range of diseases in man. The bacterium readily acquires resistance against all classes of antibiotics by one of two distinct mechanisms: mutation of an existing bacterial gene or horizontal transfer of a resistance gene from another bacterium. Several mobile genetic elements carrying exogenous antibiotic resistance genes might mediate resistance acquisition. Of all the resistance traits S aureus has acquired since the introduction of antimicrobial chemotherapy in the 1930s, meticillin resistance is clinically the most important, since a single genetic element confers resistance to the most commonly prescribed class of antimicrobials--the beta-lactam antibiotics, which include penicillins, cephalosporins, and carbapenems.

The number of Staphylococcus aureus bacteremia cases has increased annually in Denmark during the latest decades. This increase has occurred among older patients with hospital-acquired bacteremia. Methicillin-resistance, which earlier was a property of strains of the 83A phage-complex, has nearly disappeared, while the frequency of penicillin-resistance has increased. Today both the phage-type pattern and antibiotic resistance are nearly similar for strains from hospital-acquired and community-acquired cases. In hospitals the frequency of bacteremia after postoperative wound infections has decreased, while cases associated with intravascular catheters has increased, and these infections are now the most common cause of hospital-acquired S. aureus bacteremia. Endocarditis is most commonly found in community-acquired cases without an identified primary focus in patients between 21-50 years. Also hematogenous osteomyelitis is most common in community-acquired cases, but these infections have changed to having a high predilection for the vertebral column, and the prevalence of chronic cases has decreased.

About 5% of patients admitted to acute-care hospitals acquire nosocomial infections. A variety of factors contribute: increasing age of patients; availability, for treatment of formerly untreatable diseases, of extensive surgical and intensive medical therapies; and frequent use of antimicrobial drugs capable of selecting a resistant microbial flora. Nosocomial infections due to resistant organisms have been a problem ever since infections due to penicillinase-producing Staphylococcus aureus were noted within a few years of the introduction of penicillin. By the 1960s aerobic Gram-negative bacilli had assumed increasing importance as nosocomial pathogens, and many strains were resistant to available antimicrobials. During the 1980s the principal organisms causing nosocomial bloodstream infections were coagulase-negative staphylococci, aerobic Gram-negative bacilli, S. aureus, Candida spp., and Enterococcus spp. Coagulase-negative staphylococci and S. aureus are often methicillin-resistant, requiring parenteral use of vancomycin. Prevalence of vancomycin resistance among enterococcal isolates from patients in intensive care units has increased, likely due to increased use of this drug. Plasmid-mediated gentamicin resistance in up to 50% of enterococcal isolates, along with enhanced penicillin resistance in some strains, leaves few therapeutic options. The emergence of Enterobacteriaceae with chromosomal or plasmid-encoded extended spectrum beta-lactamases presents a world-wide problem of resistance to third generation cephalosporins. Control of these infections rests on (i) monitoring infections with such resistant organisms in an ongoing fashion, (ii) prompt institution of barrier precautions when infected or colonized patients are identified, and (iii) appropriate use of antimicrobials through implementation of antibiotic control programs.

Resistance to antimicrobials in bacteria results from either evolution of "new" DNA or from variation in existing DNA. Evidence suggests that new DNA did not originate since the use of antibiotics in medicine, but evolved long ago in soil bacteria. This evidence is based on functional and structural homologies of resistance proteins in human pathogens, and resistance proteins or physiological proteins of soil bacteria. Variation in existing DNA has been shown to comprise variations in structural or regulatory genes of the normal chromosome or mutations in already existing plasmid-mediated resistance genes modifying the resistance phenotype. The success of R-determinants in human pathogens was due to their horizontal spread by transformation, transduction and conjugation. Furthermore, transposition has enabled bacteria to efficiently distribute R-determinants between independent DNA-molecules. Since the genetic processes involved in the development of resistance are rare events, the selective pressure exerted by antibiotics has significantly contributed to the overall evolutionary picture. With few exceptions, experimental data about the role of antibiotic usage outside human medicine with respect to the resistance problem in human pathogens are missing. Epidemiological data about the occurrence of resistance in human pathogens seem to indicate that the major contributing factor to the problem we face today was the extensive use of antibiotics in medicine itself.

The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in the anterior nares and axilla was studied in 920 non-hospitalized subjects: 350 drug addicts, 350 individuals presenting at a hospital emergency room for various reasons, and 220 hospital health care workers. S. aureus was isolated from 105 (11.4%) subjects, in six (6.3%) of whom the isolates were methicillin-resistant. The isolation rate of the organism and the prevalence of resistant strains in the different subgroups were, respectively: drug-addicts, n = 32 (9.1%), n = 2 (6.9%); emergency room patients, n = 36 (10.2%), n = 1 (3.2%); and hospital health care workers, n = 37 (16.8%), n = 3 (8.5%). Our findings suggest that MRSA remains uncommon in the community, while the prevalence of S. aureus carriage (including methicillin-resistant strains) in hospital personnel is quite similar in divergent geographical areas.

For a 13-year period (1978 through 1990), oxacillin-resistant (MIC, greater than 4 micrograms/ml) Staphylococcus aureus (ORSA) strains were collected from Clinical Center (National Institutes of Health) patients and patients from five other U.S. hospitals. From Clinical Center patients, 251 of 253 isolates (99%) were bacteriophage typed as phage group III. Five other hospitals contributed 203 ORSA strains, of which 188 (93%) were group III. The group III ORSA strains predominantly included a characteristic core pattern of phages, 7/47/53/54/75/77. For the low-level (borderline) oxacillin-resistant strains (MIC, 2 to 4 micrograms/ml), amoxicillin-clavulanic acid combination (Augmentin) testing disclosed 62 hyper-beta-lactamase producers, of which 59 (95%) were of a separate, distinct S. aureus strain, with the phage pattern 92/94/96/292/D-11 (group V). Thus, ORSA and hyper-beta-lactamase producing S. aureus are distinct epidemic strains.

The in vitro susceptibility response of Staphylococcus aureus, Klebsiella pneumoniae, Escherichia coli, Proteus mirabilis and Pseudomonas aeruginosa to a set of antibiotics was investigated in a survey comprising 19,380 positive cultures over a period of 5 years in a large hospital environment. Four out of the five species (P. aeruginosa being the exception) presented a species-specific, drug-independent, rhythmic variation of their level of susceptibility to several antibiotics over the time of the study. The species-specific rhythmic responses were further characterized by spectral analysis, autocorrelation and cross-correlation functions. Through this analysis it was possible to rank the species according to their main period of oscillation. The longest period of oscillation was detected for S. aureus (38 months). K. pneumoniae and E. coli presented intermediate values (25 and 23 months respectively), and P. mirabilis the shortest period of oscillation (11 months). Species displaying long periods of oscillation tended to present very low levels of susceptibility, while species displaying short periods of oscillation usually presented the highest levels of susceptibility observed. Although some hospital environmental factors, such as drug consumption, were also analyzed, no correlation was found between them and the in vitro bacterial cyclic responses to antibiotics.

Conventional bacteriophage typing was combined with ribotyping in the analysis of methicillin and ciprofloxacin resistant Staphylococcus aureus strains isolated in increasing frequency since the introduction of the new 4-quinolones as therapeutic agents in the Tel-Aviv Medical Center. Whole-cell DNA was digested with EcoRI and HindIII restriction endonucleases. Agarose gel electrophoresis, Southern blotting, and hybridization by biotinylated probe DNA coding for ribosomal RNA revealed 7 to 14 bands. Analysis of the patterns established a single DNA type in EcoRI as well as in HindIII digests for all strains except one. Control strains from other sources differed in their band patterns. Bacteriophage typing confirmed the results of DNA typing. Thus, the frequent occurrence of staphylococcal isolates resistant to 4-quinolones in the hospital was not due to mutational development of resistance in many strains, but to the spread of a resistant strain.

136 methicillin-resistant strains of Staphylococcus aureus recovered from hospitalized patients in 18 countries were characterized by electrophoretic mobilities of three types of esterases. These were defined by their ranges of activity toward five synthetic substrates and their resistance to di-isopropyl fluorophosphate. Fourteen zymotypes were distinguished. Two, designated as 6 and 14, were found in 53 and 50 strains, respectively. Genetic diversity coefficients were lower for strains from France and from other European countries (H = 0.47 and 0.53, respectively) than for strains from North America (H = 0.79). On the basis of electrophoretic polymorphism of esterases, our work provides evidence that methicillin-resistance is expressed in genetically different strains. Variations in esterase electrophoretic pattern within methicillin-resistant strains of S. aureus can make a significant contribution to the study of their epidemiology.

Of 27 unique clinical isolates of methicillin-resistant Staphylococcus aureus, only 4 were homogeneously resistant, and all 4 produced little or no beta-lactamase. Among heterogeneously resistant strains, those most resistant to beta-lactam antibiotics produced the most beta-lactamase. Similar genes may regulate production of the low-affinity penicillin-binding protein and beta-lactamase.

Most of the aminoglycoside resistant Staphylococcus aureus strains isolated in France are resistant to all the antibiotics belonging to this family. Two aminoglycoside-modifying enzymes were detected in the wild-type strains studied: an APH3'III and an AAC6'-APH2". These strains also carry two types of streptomycin resistance: high-level resistance due to chromosomal mutation(s) affecting ribosome affinity and low-level resistance, the mechanism of which was not characterized. All the aminoglycoside resistance genes were located on the chromosome. DNA fragments of 1.5 and 1.95 kb carrying the aphA and aacA genes, respectively, were isolated, by cloning, from the cellular DNA of a clinical isolate. When these genes were introduced into Escherichia coli and Bacillus subtilis strains, the enzymes synthesized were indistinguishable from those produced by the S. aureus strains. When the cellular DNAs of wild-type and resistant strains were hybridized with the cloned fragments, sequences homologous to the fragment carrying the aphA gene were found to be located at the same chromosomal site, while those hybridizing with the fragment carrying the aacA gene were at different chromosomal sites.

An outbreak of gentamicin-methicillin-resistant Staphylococcus aureus (gentamicin-resistant MRSA) which occurred during 1983-84 at the Whittington Hospital, London, is described. This involved a total of 40 patients and staff and was largely confined to a urology ward. Seventeen of the patients had catheter-associated infections, 11 had wound infections and five had simultaneous infections of the urine and wound. Six members of staff and one patient were colonized with gentamicin-resistant MRSA. Vigorous and extensive infection control methods were carried out together with the use of a topical antibacterial nasal cream, mupirocin (pseudomonic acid). Seventeen patients and staff were treated with this agent and all were cleared of the original gentamicin-resistant MRSA. Sixteen patients who were not treated with mupirocin were discharged home still carrying the epidemic strain. Sporadic cases of gentamicin-resistant MRSA still occur at infrequent intervals.

A substantial portion of recently isolated, multiply-resistant Staphylococcus aureus was shown to carry R-determinants in plasmids. Some of these plasmids were small (3 Mdal) and carried only one R-marker. Others were 18-36 Mdal in size and carried two or more R-determinants. Several of the larger plasmids could be transferred by a conjugation-like process. The location of R-markers on transposable DNA sequences also was observed. Transposition as well as stable integration of R-plasmids into the chromosome can explain the frequent observation of chromosomal location of resistance. Chromosomal resistance might be an advantage for an organism frequently exposed to antibiotics. Molecular evidence suggests that methicillin resistance resides on additional chromosomal DNA. The organization of staphylococcal genomes as well as efficient transfer processes explain the genetic versatility of Staph. aureus, which has resulted in the development of multiply-resistant strains.

Semiautomated rapid broth elution (Autobac Multi-Test System; General Diagnostics, Div. Warner-Lambert Co., Morris Plains, N.J.) and disk diffusion tests were compared with an agar dilution breakpoint method to determine the antibiotic susceptibility of 147 methicillin-resistant Staphylococcus aureus isolates from our hospital. Although the disk diffusion method, in general, correlated well with the agar dilution tests, the overall agreement of the Autobac tests with agar dilution tests was only 79%, with many very major discrepancies occurring with clindamycin (88%), gentamicin (33%), and methicillin (15%). When we used a 10-fold higher inoculum for the Autobac tests, all isolates were shown to be resistant to methicillin, but significant numbers of major and minor discrepancies occurred with chloramphenicol, fusidic acid, and neomycin. The majority of isolates were shown to belong to three biotypes, distinguishable by lactose fermentation, lipolysis, hemolysis, and pigment production. The antibiotic susceptibility profile of one biotype was found to be markedly different from those of the other biotypes and contained a high incidence of clindamycin susceptibility and neomycin, gentamicin, and kanamycin resistance. In contrast, the other two biotypes had a high incidence of clindamycin, gentamicin, and kanamycin resistance and neomycin susceptibility and accounted for most of the very major discrepancies in the clindamycin and aminoglycoside tests. In these methicillin-resistant S. aureus strains, discrepancies possibly may arise from partial expression of methicillin resistance, dissociated or inducible clindamycin resistance, and instability of gentamicin resistance.

Resistance to beta-lactams may be difficult to recognize. This is due to the difficulty in detecting these resistances, when the routine tests performed in diagnostic laboratories are interpreted in the usual manner. Since failure to recognize this type of resistance may have serious consequences for the patient, it is essential that it be detected when present. For the detection of methicillin resistance of Staphylococcus aureus a standardized method using either a medium containing 5% NaCl or a low incubation temperature is advocated. Methicillin resistance of S. epidermidis can only be recognized reliably by means of a quantitative test and incubation for 42-48 h. Resistance of Haemophilus influenzae to ampicillin may be intrinsic or it may be caused by a TEM beta-lactamase; a beta-lactamase test should be used to detect the latter type of resistance. Inducible cephalosporinase may be responsible for the rapid development of resistance of some bacterial species to cefamandole, even during therapy. If a stable beta-lactamase production is attained by mutation, resistance to other beta-lactams will usually be present as well. Routine induction tests should be performed for all isolates of species of Enterobacter, Serratia, Citrobacter and Proteus, indole-positive. The same type of 'hidden' resistance may be present in Pseudomonas aeruginosa, with regard to cefotaxime and other third-generation cephalosporins. Beta-lactamase-positive Neisseria gonorrhoeae can easily be recognized by a beta-lactamase test. In addition, the results of diffusion tests allow one to distinguish between beta-lactamase-positive and beta-lactamase-negative strains. Recognition of those strains of N. gonorrhoeae having a decreased susceptibility to penicillin is only possible when well-standardized quantitative tests are used.

The antibacterial activity of cefamandole was evaluated against 120 methicillin-susceptible and methicillin-resistant Staphylococcus aureus and coagulase-negative staphylococcal strains, using both large (10(8) CFU per ml) and small (10(4) CFU per ml) inocula. Cefamandole appeared superior to cephalothin against methicillin-resistant strains, displaying at 10 micrograms/ml a bacteriostatic and for some strains a bactericidal action against a large inoculum of bacteria.

Three different systems were used to test 236 isolates of methicillin-resistant Staphylococcus aureus in an attempt to ascertain if more than one strain is responsible for the current problem of cross-infection by this organism in N.S.W. hospitals. The biochemical tests used were of little assistance. Phage typing, using the Basic International Set of typing phages at 100 x routine test dilution (RTD), provided evidence of the presence of several different strains. Phage type 83A/85/95/90/88 was the typing pattern of the predominant strain and the nest most frequent group was not typable. These results were often difficult to read. Five new phages were therefore isolated and found to be valuable as they produced easily identifiable patterns at RTD.

Although several outbreaks of methicillin-resistant Staphylococcus aureus (MRSA) infections have been reported in recent years, the geographic distribution and frequency of MRSA infections in American hospitals is unknown. We conducted a questionnaire survey to determine the magnitude of the problem. Data from 261 hospitals were included in the survey. MRSA were reported by 145 hospitals located in 36 states. Large hospitals reported these organisms significantly more often than small hospitals (p less than .001). University hospitals reported MRSA more often than community or community-teaching hospitals (p less than .001 and p less than .005, respectively). The number of hospitals reporting MRSA increased from 24 in 1975 to 112 in 1980 (p less than .001. Our data suggest that MRSA are widely distributed geographically and that the number of hospitals with these organisms has increased dramatically since 1975.

A methicillin/aminoglycoside-resistant strain of Staphylococcus aureus (MARS) was likely introduced by transfer of a patient from another hospital. Over the next year, 20 other patients were colonized or infected with MARS of the same phage type, although antibiograms varied. Affected patients usually had serious underlying disease and were in intensive care units. Vancomycin therapy was frequently delayed and MARS may have contributed to the death of some patients. The mode of spread was not definitively delineated, but two nurses were found to be colonized. Institution of isolation procedures was difficult, but the problem gradually waned. Susceptibility testing showed vancomycin to be the most active agent. Synergy studies showed no consistent effect of combining methicillin with an aminoglycoside. This experience illustrates the problem of MARS spread between hospitals and wards, the need for institution of effective control measures, and consideration of early empiric use of vancomycin.

The activity of new cephalosporins was examined using 60 coagulase-positive, methicillin-resistant staphylococcus strains and compared with that of cefalothin, methicillin and oxacillin. The results showed cefamandole to be the most effective of the new cephalosporins, its activity being comparable to that of cefalothin; cefoxitin was comparable to oxacillin and methicillin. Cefuroxime, cefoperazone, cefotaxime and moxalactam were ineffective against staphylococci. Varying degrees of hetero-resistance to all antibiotics of the beta-lactam group were found. A synergistic effect was obtained with subinhibitory concentrations of cefoxitin in combination with cefamandole or cefuroxime. In an investigation of routine determination of methicillin resistance by the diffusion method, it was found that resistance can be ascertained reliably with the technique of both the German Industrial Norm (DIN), which has not yet been officially introduced, and the technique of the American Food and Drug Administration (FDA) using a methicillin disc (5 microgram) or an oxacillin disc (1 microgram) and an incubation temperature of 35 degree C. The possibility of using beta-lactam antibiotics in infections caused by methicillin-resistant staphylococci is discussed.

A methicillin-resistant strain of Staphylococcus aureus (MRSA, phage type 84/85) was introduced into City of Memphis Hospital by a burn patient who had recently been treated for MRSA bacteremia in another institution 500 miles distant. Despite prompt recognition of the problem and institution of isolation procedures, six other patients developed secondary colonization during the ensuing six months, and five of these experienced clinically significant infections with MRSA. Three of the patients originally infected with MRSA, as well as two additional patients, subsequently developed colonization with staphylococcal strains of phage type 84/85 that were resistant to both methicillin and gentamicin (MRGRSA). Spread of the staphylococcal strains was most likely accomplished primarily via passive transfer from person to person. The hydrotherapy unit, which became contaminated with both MRSA and MRGRSA, may have played a secondary role. As illustrated by this outbreak, patients carrying potentially dangerous bacterial strains should be identified and informed of the problems posed by such carriage. It may be imprudent to admit such patients to hospitals that are free of the potential pathogen. The outbreak described here exemplifies a number of potential problems associated with control of nosocomial staphylococcal infections: (a) interhospital spread of methicillin-resistant strains; (b) secondary patient-to-patient intrahospital spread; and (c) emergence of even more resistant strains, possibly associated with selective pressures exerted by widespread use of broad-spectrum antimicrobial agents.

Fifteen patients with bacterial endocarditis were treated with vancomycin between 1967 and 1976. The indications for vancomycin therapy were penicillin-cephalosporin allergy in six patients, antibiotic resistant bacteria in six, initial therapy in one and culture-negative endocarditis in two. The causative microorganisms were Staph. epidermidis (four patients), Staph. aureus (two patients), diphtheroids (four patients), viridans streptococci (two patients) and enterococci (one patient). Minimum inhibitory concentrations of vancomycin for these organisms ranged from 0.8 to 3.1 micrograms/ml. The patients received vancomycin for two to 10 weeks (mean five weeks). Cure was achieved in 13 patients, including six with prosthetic valve endocarditis (PVE). Two patients had a relapse of PVE and cultures of blood or heart valve were positive within two months of vancomycin therapy. Vancomycin serum levels did not exceed 50 micrograms/ml, and no serious drug toxicity was encountered in any patient. Three patients had minimal audiogram changes beyond the social hearing range. One patient had mild phlebitis and a rash, and one patient had a transient leukopenia. Vancomycin is an effective nontoxic antibiotic in patients with endocarditis when penicillin or cephalosporin therapy is not appropriate.

In the period 1969-1974 a decreasing number of Danish hospital departments recorded epidemic occurrences of Staphylococcus aureus strains, and the local spread of strains was less extensive. Multiple-resistant strains of the 83 A complex were succeeded by type 94 strains, resistant to penicillin only. The level of methicillin resistance fell from 19 to 6%. 12% of methicillin resistant strains are now sensitive to streptomycin and/or tetracyclines; they do not represent a few clones, but belong to a wide spectrum of phage types. The changes follow a reduction in the consumption of streptomycin and tetracyclines not of methicillin or other penicillins. As a contrast to the general reduction of combined resistance to streptomycin and tetracyclines, the strains in dermatological departments, where tetracyclines are commonly used, maintain a high degree of resistance to tetracyclines alone.

During the years 1966-1974, 167,297 strains isolated from 167,297 patients or staff members in Danish hospitals were registered at Staten Seruminstitu. All the strains were phage-typed and examined for production of a 'Tween'-80-splitting enzyme and resistance to mercuric chloride. 158,236 strains were examined for resistance to antibiotics. Since 1968, a steep decrease in the number of strains resistant to three or more antibiotics (multiple-resistant) and in strains of the 83A complex was noticed. In recent years an increase in strains belonging to phage-group I and in those referred to as miscellaneous and non-identified was registered. The increase in the non-typable strains might be explained by the shift of the concentration of the typing phages from 1000 X RTD to 100 X RTD. It is concluded that at least two factors may have contributed to the reduction of the multiple-resistant strains: an altered antibiotic policy, restricting the use of streptomycin and tetracyclines, and an improved hospital hygiene, diminishing the spread of identical strains within the various departments. However, it is emphasized that the consumption of methicillin is still increasing.

Anaerobic bacteria recovered from airway-related infections were tested by agar dilution against selected penicillins and cephalosporins available for oral administration. Against 136 isolates, penicillins G and V showed comparable activity, particularly when pharmacological differences were considered. Although many isolates were exquisitely susceptible to the penicillins, only 55% of the Bacteroides species and 72% of all isolates were inhibited at 0.5 mug of penicillin G per ml. Results for penicillin V at 1 mug/ml were similar (59 and 73%). The two cephalosporins were more active at achievable levels, inhibiting 94 to 95% of Bacteroides and 95 to 96% of all isolates at 8 mug/ml. These levels represent approximately 50% of the reported peak serum levels after oral administration of 625 mg of the penicillins and 500 mg of the cephalosporins. Dicloxacillin and nafcillin were tested against 50 isolates. The two were comparably active on a weight basis; dicloxacillin was more active when pharmacological differences were considered, but did not match the other penicillins or the cephalosporins.