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Analysis of antimicrobial drug resistance of Staphylococcus aureus strains by WHONET 5: microbiology laboratory database software.

OBJECTIVES: To analyze our hospital laboratory microbiological data by using WHONET 5-Microbiology laboratory database software-, and to acquire information about antimicrobial resistance of Staphylococcus aureus strains among every ward. MATERIALS AND METHODS: The database of Staphylococcus aureus strains had been brought to our hospital microbiology laboratory from every ward in our hospital from September 2001 till December 2002. Analysis was performed under the condition as one isolate per one patient. Starting of "resistance profile" analysis in WHONET 5 and analyzing the microbiological laboratory testing reports for every ward. We chose Oxacillin, Levofloxacin, Erythromycin and Gentamicin as the antimicrobials that need to be investigated for resistance. We evaluated the monthly transition of resistance ratios with regard to the specific wards that have the moving lines of inpatients in order to verify the hypothesis that resistant strains may be carried from ward to ward along the moving lines of inpatients. RESULTS: The data of 2,113 Staphylococcus aureus strains were accumulated and analyzed. Overall Oxacillin resistance ratio in our hospital was 65.7%. The ward of the smallest Oxacillin resistance ratio was Pediatrics/Ophthalmology ward. The ratios of Oxacillin resistant were varied as from 67.9% to 96.7% regardless the categories of wards such as internal medicine or surgery. Multi-resistant MRSA strains were overwhelmingly dominant in the wards of surgery. The ratios of the Gentamicin sensitive strains that were resistant to Oxacillin were high over the every ward. The moving lines of inpatients existed between ICU/CCU ward and three rear wards. Two rear wards whose Oxacillin resistance changes were reflected to those of ICU/CCU, but one rear ward was not. CONCLUSION: Variation of resistant degree among wards were very obvious and large. We could survey the wards where patient-to-patient transmission of resistant organisms might occur along the moving lines of inpatients. WHONET 5 will be recognized as an analysis and surveillance tool for every infection control team to survey the suspicious wards.

Anti-Bacterial Agents↗

Phylogenetic analyses of Staphylococcus based on the 16S rDNA sequence and assignment of clinical isolates from animals.

The nucleotide sequences of the 16S rDNA in 17 strains of 16 taxa of the genus Staphylococcus were determined. The sequences were compared phylogenetically together with the gene sequences of 10 (including 7 other species) Staphylococcus species retrieved from the DNA Data Bank of Japan. Although the primary and secondary structures of most of Staphylococcus species were very similar (homology values 96.4% or more) except for S. caseolyticus MAFF 911387T (homology values 95.4% or less), the 23 staphylococcal species were divided into 10 groups based on similarity, evolutionary distance and phylogenetic tree analysis. Nucleotide stretches in several variable domains in the 16S rDNA sequences appeared to be specific for the bacterial groups or the species. By comparing such characteristics in the sequence and phylogenies of 5 staphylococcal clinical isolates from bovine mastitis, canine and feline pyoderma, and feline urogenital syndrome with the information obtained in this study, the species level of each organism was identified.

Animals↗

Development of a PCR test for the identification of Staphylococcus intermedius based on the 16S rDNA sequence.

The development of a PCR assay based on the 16S ribosomal RNA gene (rDNA) sequence was carried out for the identification of Staphylococcus intermedius. Sixty-six strains of S. intermedius, 70 of Staphylococcus aureus and 2 of Staphylococcus hyicus were examined for the assay. The 16S rDNA, of which the PCR target fragment makes up 901 bp corresponding to the sequence data of the gene, was detected in all strains of S. intermedius, but it was not detected in any strains of either S. aureus or S. hyicus. These results suggest that the PCR allows a simple and precise identification of S. intermedius.

Coagulase↗

Nasal carriage of methicillin-resistant and methicillin-susceptible Staphylococcus aureus in nursing home residents in Bolu, Turkey.

BACKGROUND: This study aimed (a) to provide information on methicillin-resistant (MRSA) and methicillin-susceptible (MSSA) Staphylococcus aureus colonization ratio in residents of nursing homes; (b) to determine the effect of hand-washing education given to nursing home residents and employees on nasal carriage of Staphylococcus aureus (NCSA) and (c) to obtain probable risk factors for MRSA colonization of residents in two nursing homes. METHODS: Seventy-nine volunteers (24 females and 55 males) from Bolu nursing homes were included in the study. Nasal samples were taken with sterile swabs from the anterior nares. Staphylococcus aureus strains were identified using classical methods and genotyping of methicillin resistant strains was done using Arbitrary Primed PCR (AP-PCR). Antibiotic susceptibilities were determined by disk diffusion methods according to NCCLS standards. After first nasal samples were taken, all employees and residents of nursing homes were educated about the methods of hand hygiene over two days. RESULTS: With hand-washing education, the decrease of NCSA rate (initially 43%; after education, 21%) was significant (p < 0.05) while decrease of MRSA carriage (initially 5%, after education 1%) was not (p > 0.05). MRSA carriage was significantly correlated with presence of skin lesions, prior hospitalization within the last six months, and antibiotics usage within the last six months. AP PCR results suggested that residents' carriage of MRSA was the result of the same source. CONCLUSION: MSSA and MRSA colonization rates were found to be 38% and 5% in nursing homes, respectively. These ratios can decrease with simple precautions like hand-washing after a short education period

Aged↗

Isolation of Staphylococcus schleiferi from healthy dogs and dogs with otitis, pyoderma, or both.

OBJECTIVE: To determine the frequency of isolation and susceptibility patterns of Staphylococcus schleiferi from healthy dogs and dogs with otitis, pyoderma, or both that had or had not received antimicrobial treatment. DESIGN: Prospective study. ANIMALS: 50 dogs. PROCEDURE: Dogs were allocated to 1 of 4 groups: healthy dogs (n=13), dogs without otitis but with pyoderma (10), dogs with otitis but without pyoderma (11), and dogs with otitis and pyoderma (16). Bacteriologic culture of ear swab specimens was performed in all dogs. Bacteriologic culture of skin swab specimens was also performed in dogs with concurrent pyoderma. Isolates were identified as S schleiferi subsp schleiferi or S schleiferi subsp coagulans on the basis of growth and biochemical characteristics. RESULTS: S schleiferi was not isolated from any dogs with pyoderma only. Staphylococcus schleiferi subsp schleiferi was isolated from the ears of 2 healthy dogs, and the skin and ears of 2 dogs and the skin of 1 dog with otitis and pyoderma. Staphylococcus schleiferi subsp coagulans was isolated from the ears of 3 dogs with otitis only, and the ears of 6 dogs and the skin of 2 dogs with otitis and pyoderma. One of the S schleiferi subsp schleiferi isolates from ears, 2 of the S schleiferi subsp coagulans isolates from ears, and 1 of the S schleiferi subsp coagulans isolates from the skin were resistant to methicillin. One methicillin-resistant isolate from the ears and 1 from the skin were also resistant to fluoroquinolones. CONCLUSIONS AND CLINICAL RELEVANCE: S schleiferi subsp schleiferi was detected in healthy dogs and dogs with otitis and pyoderma. Methicillin-resistant and -susceptible S schleiferi subsp schleiferi and S schleiferi subsp coagulans were detected as the predominant organisms in dogs with otitis.

Animals↗

The occurrence of Staphylococcus aureus on a farm with small-scale production of raw milk cheese.

In recent years, the small-scale production of raw milk products has increased in Norway, and there is some concern that such foods may pose a risk of staphylococcal food poisoning to consumers. The aim of the study was to evaluate potential sources of contamination of raw milk cheese with Staphylococcus aureus on a bovine dairy farm with small-scale production. Samples for bacteriological analyses (n = 144) were collected from the animals, the environment, processing equipments, from humans, and from cheeses at various stages of production. Staphylococcus aureus was isolated from 10 of 11 cows, the farmer, equipment, the environment, and the cheese. Seventy-five Staph. aureus isolates were genotyped by pulsed-field gel electrophoresis, tested for enterotoxin (SE) production by reversed passive latex agglutination, for SE genes by multiplex polymerase chain reaction, and for penicillin resistance by the cloverleaf method. Five different pulsotypes were identified and SE gene fragments were identified in 11 isolates, but no isolates produced SE or were penicillin resistant. Staphylococcus aureus was found throughout the farm, and appeared to be spread with the milk to the environment, equipment, and to products. One pulsotype dominated and was identified from most sample sites on the farm. Raw milk products are vulnerable to contamination with Staph. aureus. Strategies to reduce the occurrence of Staph. aureus in bulk milk are of particular importance on farms where milk is used for raw milk products.

Animals↗

Novel immunofluorescent technique for identification of ovine immunoglobulins and other potential opsonins binding to live Staphylococcus aureus.

A qualitative in vitro technique has been developed for identification of potential opsonins (immunoglobulins G1, G2, A, M; the third component of ovine complement C3, and fibronectin) of sheep origin, binding to cell walls of viable Staphylococcus aureus. The technique uses monospecific antisera to these ovine proteins. The antisera are conjugated to FITC-protein A such that the protein A-binding sites in the Fc region of the immunoglobulin molecules are occupied with FITC-protein A complexes and are prevented, therefore, from binding to protein A in the staphylococcal cell wall. The technique is highly specific and sensitive, and once conjugated monospecific antisera are prepared, many tests can be done in a short time. Staphylococcus aureus incubated in samples of milk whey showed variable binding of immunoglobulins G1, G2, and M; immunoglobulin A was bound from only one sample of milk whey, and the third component of ovine complement C3- and fibronectin binding were not detected. Most samples of blood serum and colostral whey produced binding of immunoglobulins G1, G2, A, and M, and the third component of ovine complement C3 to the Staphylococcus aureus cell surface; fibronectin binding was found in serum but not in colostral whey. Washings from involuted mammary glands invariably produced binding of all immunoglobulins but not of the third component of ovine complement C3 or fibronectin.

Animals↗

Germicidal teat dip in a herd with low prevalence of Streptococcus agalactiae and Staphylococcus aureus mastitis.

Effectiveness of an iodophor teat dip in reducing new intramammary infection and clinical mastitis was tested in a herd free of Streptococcus agalactiae and with a low prevalence of Staphylococcus aureus infections. In this 6-mo study with an average of 152 lactating cows on trial on sampling dates, right rear and left front teats were dipped after each milking while right front and left rear teats were undipped controls. Teat dipping significantly reduced new infections by Staphylococcus aureus, streptococci (other than Streptococcus agalactiae), coagulase-negative staphylococci, and Corynebacterium bovis and reduced clinical cases caused by infections established during the trial. However, the dip did not reduce new infections or clinical mastitis caused by coliform bacteria. Therefore, use of a germicidal dip is recommended for herds with a low prevalence of Streptococcus agalactiae and Staphylococcus aureus, but the practice should not be expected to control coliform mastitis.

Administration, Topical↗

Evaluation of protein A and a commercial bacterin as vaccines against Staphylococcus aureus mastitis by experimental challenge.

Protein A and a commercial staphylococcal bacterin were evaluated by experimental challenge with Staphylococcus aureus (ATCC 29740). Thirty cows in first lactation were in three treatment groups, protein A, bacterin, and nonvaccinated controls. Studies were through three lactations and included bacteriological and cytological analyses of quarter milk samples. Rate of intramammary infection with Staphylococcus aureus was similar for vaccinated and unvaccinated cows. Rates of spontaneous cure within each lactation were significantly higher for vaccinated cows. For all three lactations, spontaneous cure rates were 83, 73, and 47% for protein A, bacterin, and control cows. Somatic cell counts were significantly lower for vaccinated cows for quarters infected with Staphylococcus aureus, but no differences were demonstrated for milk production by lactation. Incidence of clinical mastitis was higher in unvaccinated cows, but too few developed for a valid comparison.

Animals↗

Growth responses of Staphylococcus aureus and Streptococcus agalactiae to Corynebacterium bovis metabolites.

Staphylococcus aureus and Streptococcus agalactiae growth responses to metabolites of Corynebacterium bovis cultured in media containing polyoxyethylenesorbitan monolaurate, monooleate, or trioleate and milk were determined. Filter sterilized metabolites of 48-h C. bovis cultures in synthetic media were added to cultures of Staph. aureus and Strep. agalactiae. Staphylococcus aureus and Strep. agalactiae were inoculated into 12-h C. bovis milk cultures. Growth responses of Staph. aureus and Strep. agalactiae were not affected by C. bovis metabolites of synthetic media. Staphylococcus aureus growth was inhibited during logarithmic and stationary phases in milk containing mixed cultures of C. bovis compared with growth in pure Staph. aureus cultures. Streptococcus agalactiae growth curves were similar in pure and C. bovis mixed cultures. Fatty acid compositions were not different in sterile milk and milk containing bacterial cultures. Growth responses of Staph. aureus and Strep. agalactiae were not related to concentration of C. bovis metabolites or fatty acid content of media in which C. bovis were cultured.

Corynebacterium↗

Colonization by Staphylococcus aureus on chapped teat skin: effect of iodine and chlorhexidine postmilking disinfectants.

The abilities of common postmilking teat disinfectant solutions and a teat skin ointment to retard Staphylococcus aureus colonization and promote healing of chapped skin were studied. Ten Holstein cows, free of S. aureus intramammary infections and mammary epidermal colonization, were used. All teats were experimentally chapped with 1N sodium hydroxide and dipped in S. aureus broth culture of 5 x 10(6) cfu/ml after three consecutive milkings. Postmilking teat disinfectant treatments were applied, starting after the second application of S. aureus broth culture, and were continued for the next 19 milkings. Quarters of each cow received one of the following postmilking teat treatments: 1% iodine plus 10% glycerin solution; .5% chlorhexidine plus 4.87% glycerin solution; 1% chlorhexidine ointment; and no postmilking treatment. After milking and before treatment application, samples of skin swabbings of the teat end and teat side and samples of milk were collected. Daily teat skin chapping visual scores were made to assess chapping and healing. Teat skin chapping decreased with time, and the decline was similar among treatments. The concentration of S. aureus recovered from teat skin swabbing solutions decreased over time. Staphylococcus aureus were recovered at significantly lower concentrations from swabbing solutions of teats that were dipped with iodine and chlorhexidine solutions as compared with untreated teats. Staphylococcus aureus was in marginally higher concentrations on ointment-treated teats than on dipped teats. Results indicate the importance of using postmilking teat dips on chapped teats and suggest no advantage in applying a disinfectant ointment to chapped teats to reduce S. aureus colonization and improve skin healing.

Analysis of Variance↗

[The effect of silicon on the growth of Staphylococcus aureus].

Various studies on the effect of silicon on the development of chronic inflammatory diseases of the mucous membrane such as chronic sinusitis have been carried on for the past twenty years. Staphylococcus aureus is usually detected frequently in chronic sinusitis. In this study, proliferation of Staphylococcus aureus in various culture fluids with Si concentrations ranging from 10 to 410 ppm was observed through 16 hours by means of the measurement of optical density, protein concentration and viability count. As the result of this observation, higher concentration of Si in the culture fluid showed a remarkable growth accelerating effect on Staphylococcus aureus compared with normal culture fluid containing about 10 ppm of Si. High Si concentration in mucous membrane tissue was considered to help the growth of bacteria in the sinus.

Colony Count, Microbial↗

Prosthetic joint infection due to Staphylococcus lugdunensis.

Staphylococcus lugdunensis, a coagulase-negative staphylococcus, is being increasingly recognized as the cause of serious infections. We report 2 cases of total knee arthroplasty infection caused by S lugdunensis. S lugdunensis frequently produces a clumping factor that can result in a positive slide (short) coagulase test result. If the microbiology laboratory does not use the tube coagulase (long) test to confirm the slide coagulase test result, the organism may be misidentified as Staphylococcus aureus. S lugdunensis is more virulent than other coagulase-negative staphylococci and in many clinical situations behaves like S aureus, further increasing the confusion. However, S lugdunensis differs from S aureus in that it is susceptible to most antibiotics. This fact may alert the microbiology laboratory or the clinician that the isolate is likely not S aureus and prompt further testing of a specific isolate. Accurate identification of S lugdunensis isolates facilitates studies to define the epidemiology and pathogenesis of prosthetic joint infection due to S lugdunensis and delineates optimal medical and surgical therapies.

Aged↗

A 23S rDNA-targeted polymerase chain reaction-based system for detection of Staphylococcus aureus in meat starter cultures and dairy products.

A polymerase chain reaction-based system for detection of Staphylococcus aureus was developed. The system consisted of the following components: (i) selective enrichment, (ii) DNA isolation, (iii) amplification of DNA with primers targeted against the 23S rRNA gene, and (iv) evaluation of the specificity of the polymerase chain reaction by Southern hybridization and nested polymerase chain reaction. The method achieved a high degree of sensitivity and unambiguity as required for the detection of contaminants in food starter preparations. The method permitted detection of Staphylococcus aureus in preparations of meat starter cultures containing Staphylococcus carnosus either alone or in combination with lactobacilli, pediococci, and/or Kocuria varians. Detection limits were sufficiently low to show within 12 h the presence of 10(0) CFU of S. aureus in starter preparations containing 10(10) CFU of S. carnosus. The system was also applied to dried skim milk and cream. For detection without selective enrichment, a protocol was developed and permitted detection of 120 CFU of S. aureus in 1 ml of cream within 6 h. With nested polymerase chain reaction, the detection limit was decreased by one order of magnitude.

Animals↗

Inhibition of a methicillin-resistant Staphylococcus aureus strain in Afuega'l Pitu cheese by the nisin Z-producing strain Lactococcus lactis subsp. lactis IPLA 729.

Methicillin-resistant Staphylococcus aureus strains are a potential threat for food safety because foodborne illness caused by methicillin-resistant Staphylococcus aureus has been reported even though these strains were only associated with nosocomial infections until recently. This article focuses on the inhibitory effect of the nisin Z-producing strain Lactococcus lactis subsp. lactis IPLA 729 on the growth of Staphylococcus aureus CECT 4013, a methicillin-resistant strain. S. aureus was inhibited by the presence of the nisin producer IPLA 729 in buffered Trypticase soy broth, milk, and Afuega'l Pitu cheese, an acid-coagulated cheese manufactured in Asturias, Northern Spain. A reduction of 3.66 log units was observed in Trypticase soy broth at the end of the incubation period. In milk, viable counts of S. aureus were undetectable or were reduced by 2.16 log units in 24 h depending on the initial inoculum (1.8 x 10(4) and 7.2 x 10(6) CFU/ml). The staphylococcal strain was also undetected in test cheeses in which the nisin Z producer was present whereas 2 log units were detected in control cheeses at the end of ripening.

Anti-Infective Agents↗

Performance of response surface and Davey model for prediction of Staphylococcus aureus growth parameters under different experimental conditions.

The combined effect of different temperatures (7 to 19 degrees C), pH levels (4.5 to 8.5), sodium chloride levels (0 to 8%), and sodium nitrite levels (0 to 200 ppm) on the predicted growth rate and lag time of Staphylococcus aureus under aerobic and anaerobic conditions was studied. The two predictive models developed, response surface (RS) and the Davey model, provided reliable estimates of the two kinetic parameters studied. The RS provided better predictions of maximum specific growth rate, with bias factors of 1.06 and 1.31 and accuracy factors of 1.17 and 1.37, respectively, in aerobic and anaerobic conditions. The Davey model performed more accurately for lag time, with a bias factor of 1.12 and an accuracy factor of 1.49, for both aerobic and anaerobic conditions. Predictive growth models are a valuable tool, enabling swift determination of Staphylococcus aureus growth rate and lag time. These data are essential for ensuring staphylococcus-related quality and safety of food products.

Consumer Product Safety↗

Mitral and aortic valve endocarditis due to Staphylococcus lugdunensis.

Staphylococcus lugdunensis is a recently described coagulase negative staphylococcal species involved in human infections. Endocarditis caused by Staphylococcus lugdunensis has been reported rarely: fewer than 50 cases have been described so far. The infection is frequently complicated by embolic events and carries a high mortality rate. We report a case of endocarditis due to Staphylococcus lugdunensis in which the native mitral and aortic valves were infected. The bacterium was isolated on cultures from the aortic and mitral vegetations. Appropriate medical and surgical treatment led to a good outcome of the infection. At 6-year follow-up, there was no evidence of recurrence, and the patient showed good functional recovery. He was in New York Heart Association functional class I.

Aortic Valve↗

A CD-1 mouse model of infection with Staphylococcus aureus: influence of gender on infection with MRSA and MSSA isolates.

Staphylococcus aureus is an important pathogen of humans and other animals, causing bacteremia, abscessation, toxemia, and other infectious diseases. An animal model using CD-1 mice was developed to study the pathogenesis of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA). When inoculated into the CD-1 mouse model, it was shown that both MSSA isolates, (HR 78 and CSA-1) and MRSA isolates (MRSA 456 and MRSA 457) led to chronic infection of the kidney. Female CD-1 mice inoculated with MRSA 456 proved to be more susceptible to infection and mortality than their male counterparts. Castrated mice became more susceptible to infection than intact male mice, suggesting a hormonal involvement in the infection process.

Animals↗