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[Typing of methicillin-resistant Staphylococcus aureus with a specific type of resistance].

Methycillin resistant Staphylococcus aureus (MRSA) is the important causative agent of intrahospital infections (IHI) which could have epidemic character. Typization of strains is necessary for control of these infections. Fifty eight strains of MRSA with specific resistance to antibacterial agents have been isolated from patients at different clinics of our hospital. The determination of resistotype, biotype and plasmid profiles of these and control strains of Staphylococcus aureus (SA) was aimed at confirmation whether identical strains were in questions. Biotypization showed no difference between the studied and control strains while determination of the plasmid profile proved the presence of an epidemic strain which caused infection in several clinics of our hospital.

Bacterial Typing Techniques↗

Prophylactic efficacy of four antibacterial shampoos against Staphylococcus intermedius in dogs.

Prophylactic efficacy of 4 antibacterial shampoos against Staphylococcus intermedius in dogs was determined by use of a controlled quantitative technique. Ten adult Beagles were used in the study. The antibacterial agents in the shampoos were 3.0% benzoyl peroxide, 0.5% chlorhexidine acetate, 1.0% available iodine as a polyalkyleneglycol-iodine complex, and a combination of 0.5% triclosan, 2.0% sulfur, and 2.0% salicylic acid. Treated and control sites were challenge exposed with 5.30 +/- 0.10 (log10) S intermedius colony-forming units (CFU)/cm2 of skin and occluded for 5 hours. At the end of the test period, remaining bacteria were removed with a detergent cup-scrub technique and the total number of S intermedius CFU/cm2 skin was calculated for each treated and control site. Nontreated bacteria-challenged control sites yielded 5.62 +/- 0.65 S intermedius CFU/cm2 of skin. Staphylococcus intermedius recovery (CFU/cm2) from the treated sites was 0.94 +/- 0.76 for benzoyl peroxide, 1.96 +/- 1.33 for chlorhexidine acetate, 3.11 +/- 0.48 for organic iodine, and 4.69 +/- 0.23 for triclosan-sulfur-salicylic acid. Each S intermedius recovery value from the 4 treated sites was significantly (P less than 0.05) lower than that from the nontreated S intermedius challenge-exposed control site. Bacteria recovery values were also significantly (P less than 0.05) different among the 4 shampoo-treated sites. We concluded that all shampoos had significant (P less than 0.05) prophylactic activity against S intermedius over 5 hours. The shampoo containing benzoyl peroxide was determined to have the greatest efficacy among the products tested.

Animals↗

[Staphylococcus aureus sepsis in patients with hematological malignancies: increase in MRSA sepsis].

From January 1978 to August 1990, Staphylococcus aureus bacteremia (SAB) were identified in 31 patients with hematological malignancies at Jichi Medical School hospital. Mortality due to SAB was 48.4% (15/31). Of the variables analyzed, four factors were significantly associated with a poor prognosis; elderly age (p = 0.015), high granulocyte count (more than 500/microliters) (p = 0.015), presence of DIC (p = 0.011) and presence of pneumonia (p = 0.023). The incidence of methicillin-resistant SAB was 32.3% (10/31) and the first patient developed in 1985. Although not statistically significant, there was a trend of higher mortality for methicillin-resistant SAB (70%) than for methicillin-sensitive SAB (38.1%). Most strains of methicillin-resistant Staphylococcus aureus were sensitive to minocycline, chloramphenicol and vancomycin.

Adult↗

[Effect of histones on a change in the enzymatic activity of Escherichia coli and Staphylococcus aureus cells].

Histones have been found to interfere with the permeability of the cellular membranes of Escherichia coli and Staphylococcus aureus, this resulting in the liberation of 14C-amino acids, the UV-absorbing endogenous material, and the products of RNA degradation. Histones, particularly arginine-rich fractions, stimulated the RNAase activity of the bacteria. Histones inhibited the phosphatase activity of E. coli and stimulated that of Staphylococcus aureus. The ATPase activity of the cells increased upon contact with histones. These enzyme systems are presumed to be components of a mechanism whose activity increases upon the action on the cells of factors interfering with the permeability of the membranes so that low-molecular-weight compounds are liberated from the cells. It is possible that the RNAase, ATPase and phosphatase systems repair the pool of low-molecular-weight compounds, thus contributing to the survival of the bacteria.

Adenosine Triphosphatases↗

[Expression of Staphylococcus aureus enterotoxin A gene in heterologous systems].

The genomic library of Staphylococcus aureus genes on the plasmid vector pSL5 has been constructed. The library contains a 2.5 kb HindIII DNA fragment including the gene for enterotoxin A. The entA gene on the high copy number plasmids in the Escherichia coli cells deficient in proteolysis determines the synthesis of enterotoxin A in the amounts comparable to the ones in the parent strain Staphylococcus aureus FRI 722(H).

Bacillus subtilis↗

Changing phage pattern of Staphylococcus in pyoderma cases.

Staphylococcus aureus is a major pathogen for pyoderma in India. Phage pattern of bacteria gives valuable information in epidemiological studies of infection. Two hundred and two strains of Staphylococcus aureus isolated from pyoderma cases at Gorakhpur, were phage grouped and phage typed. It was found that 43.1 percent strains were not typable. Most common group was mixed phage group (23.8 percent) followed by phage group III (12.4 percent). Predominant phage types in mixed phage group was 84/81/85 and in phage group III was 84/85.

Bacteriophage Typing↗

[Isolation of Staphylococcus aureus protoplasts using lysoamidase].

The effect of the bacteriolytic enzyme preparation, lysoamidase, on Staphylococcus aureus 209P cells was studied. The protoplast formation was examined by spectrophotometric, biochemical and electron microscopic methods. Optimal conditions for isolation of S. aureus protoplasts were chosen. The susceptibility of S. aureus cells to lysoamidase depended on the culture age: the maximum effect was observed in the logarithmic growth phase. The protoplast yield was 80% when 1 M sucrose was used as an osmotic stabilizer. Lysoamidase caused local disruptures of the staphylococcus cell walls, which resulted in the formation of osmotically fragile spheroplasts and the release of protoplasts into the medium. The protoplasts obtained could retain 85-90% of the respiration activity and were able of cell wall regeneration.

Hydrolysis↗

[Determination of beta-lactamase production in strains of Staphylococcus aureus isolated from the milk of cows].

Determination of sensitivity to penicillin G by a standard disk assay diffusion method was compared with a iodometric method of test papers to determine beta-lactamase production after Jorgensen in Staphylococcus aureus strains isolated from cow's milk from different farms. Out of 179 test strains, 32 strains 17.8%) were found to be well sensitive in the diffusion test; eight of these strains (25.0%) were demonstrated to produce beta-lactamase. 54 strains (30.2%) were sensitive. 27 strains (50%) of this group produced beta-lactamase. 93 strains (51.9%) were resistant to penicillin G in the diffusion test. 86 strains (92.5%) were found to produce beta-lactamase and seven strains were negative in this test. Using the diffusion test of sensitivity in these cultures, 86 strains were sensitive (48.1%) and 93 strains were resistant (51.9%). Beta-lactamase was produced by 121 strains (67.6%) and no beta-lactamase production was recorded in 58 strains (32.4%). Differences in the results of both tests were manifest mainly in the set of strains qualified as sensitive (inhibition zone diameter 24 to 16 mm) and well sensitive (inhibition zone diameter larger than 25 mm). The results indicate that the currently performed diffusion test of sensitivity to penicillin G should be accompanied by an assay of beta-lactamase production. The iodometric method of test papers is simple, rapid and cheap and can be made in any bacteriological laboratory. The high resistance of Staphylococcus aureus strains to penicillin G documents that this antibiotic is little efficient in the treatment of mastitis of this etiology in the given region.

Animals↗

[Analysis of methicillin-cephem resistant Staphylococcus aureus (MRSA) hospital infection and toxigenicity of MRSA].

MRSA infectious diseases were often observed in our ward including general, pediatric and neurosurgery during recent two years. Following items were investigated for analysing a prevalence of the infections; 1) the frequency of MRSA in Staphylococcus aureus isolates from clinical materials, 2) the monthly number of patients with MRSA infectious diseases, 3) the biologic types and the toxigenicity of MRSA isolates from clinical materials, nasal carriers and an environmental material in the ward. The results were as follows. 1) Methicillin-cephem resistant strain was determined in 204 of 247 Staphylococcus aureus isolates (83%). 2) Monthly registration showed a trend that an increased number of the patient in one unit was followed by an increase in other units. 3) Most of MRSA isolates were classified into type II coagulase and two kinds of strains were predominantly found in isolates from clinical materials by plasmid DNA analysis. Most of MRSA isolates had capabilities of producing type C enterotoxin and toxic shock syndrome toxin-1 (TSST-1). The results suggested that the frequent MRSA infectious diseases attributed to hospital infection. Since hospital infection of virulent MRSA may cause serious infectious diseases, much concern to inhibit the spread of this organism should be required.

Bacterial Toxins↗

Failure of cephalosporins to prevent Staphylococcus aureus surgical wound infections.

Approximately 35,000 Staphylococcus aureus surgical wound infections occur annually in the United States. To investigate why S aureus causes infection despite the perioperative administration of cephalosporins, we compared 35 methicillin-susceptible isolates recovered from deep wound infections that complicated cefazolin prophylaxis (18 of 1650 patients) and cefamandole prophylaxis (17 of 3702 patients) with 64 colonizing isolates from presurgical patients. Compared with both colonizing and cefamandole-associated isolates, S aureus isolates from cefazolin-associated infections were more resistant to cefazolin by specialized assays. Staphylococcus aureus isolates that produced the A and C variants of staphylococcal beta-lactamase were associated with infections following cefazolin and cefamandole prophylaxis, respectively. These isolates hydrolyze the respective cephalosporins rapidly, suggesting that staphylococcal survival after perioperative prophylaxis may be mediated by in vivo degradation of the prophylactically administered cephalosporin. These data indicate that some S aureus wound infections occur because of deficiencies in antimicrobial effectiveness that are not detectable by routine susceptibility tests. This finding has important implications for the therapy and prevention of S aureus infection.

Cefamandole↗

[Four cases of airway infections caused by MRSA (methicillin resistant Staphylococcus aureus)].

Currently, infections caused by MRSA (methicillin-resistant Staphylococcus aureus) pose a great problem clinically. We present 4 patients with MRSA infections experienced by us. In these patients the infection was localized in the trachea and main bronchus. The first patient was a 62-year-old man. After undergoing operation for early gastric cancer, he had septic shock and was admitted to our center. The second was a 60-year-old man. After he underwent operation for advanced gastric carcinoma at another hospital septicemia developed due to suture failure and he was admitted to our center. The third was a 38-year-old woman who was admitted to our center because of grades II degrees-III degrees burns on 75 to 80% of her body surface area. The fourth was a 60-year-old man who was admitted to our center because of rupture of an aneurysm of the abdominal aorta. It is assumed that MRSA has quite different characteristics from the usual MSSA (methicillin-sensitive Staphylococcus aureus) in that it produces a new penicillin-bound protein (PBP-2') within cells. Thus, from the fiberoptic bronchoscopy findings of our own cases it is considered that there may be cases in which the observed lesion is localized in the central airway alone, without involvement of the segmental bronchi. We believe it necessary to take some prompt measures under a suspicion of airway infection caused by MRSA in the following cases: (1) compromised hosts under tracheal intubation, (2) patients who are under treatment with second or third generation cephalosporins, and (3) patients with production of bloody sputum, and (4) endotoxin-positive patients.

Adult↗

The adherence of Staphylococcus aureus to human corneocytes.

Staphylococcus aureus has a peculiar ability to colonize the skin of patients with atopic dermatitis. We examined the possibility that this might be due to a specific ability of this pathogenic staphylococcus to adhere to atopic stratum corneum. We used an in vitro model to show that S aureus does have an unusual ability to adhere to atopic corneocytes when compared with corneocytes obtained from patients with other cutaneous diseases, including psoriasis. Protein A--a component of the staphylococcal cell wall--may be responsible in part for this adherence phenomenon. This trait did not extend to the other gram-positive bacteria tested.

Cell Division↗

On chronic prostatitis with special studies of Staphylococcus epidermidis.

Patients with chronic prostatitis have an increased number of white blood cells in expressed prostatic secretion. Two groups can be separated, one is characterized by uropathogenic bacteria in expressed prostatic secretion and recurrent urinary tract infections, chronic bacterial prostatitis. In this group an immune response to the bacteria has been demonstrated. Patients belonging to the other group, non-bacterial prostatitis, have similar symptoms. Many harbour Gram-positive bacteria in a high number, often Staphylococcus epidermidis in expressed prostatic secretion. This bacteria is usually not considered in prostatitis in spite of extreme high numbers. The etiology of non-bacterial inflammations is thus unknown. Forty-three per cent of the patients with chronic prostatitis had Gram-positive bacteria and 13% had Gram-negative in expressed prostatic secretion. Forty-four per cent of patients referred with symptoms of prostatitis did not have any aerobic bacteria at the prostatic level in sufficient number for the diagnosis bacterial prostatitis according to Meares and Stamey and form thus a third group. Antibiotic treatment of patients with non-bacterial prostatitis reduced symptoms but also changed the bacterial flora in urethral and prostatic secretion in such a way that uropathogens were found after treatment. In a group of patients an immunologic response to Staph. epidermidis was searched for by measuring complement components (C3c, C4c) as well as ceruloplasmin in serum and immunoglobulins (IgA, IgG) in seminal plasma. A specific ELISA method to estimate antibodies in serum against Staph. epidermidis was tested. No specific pattern separated patients from controls or patients with Gram-negative bacteria from patients with Gram-positive bacteria. Staphylococcus saprophyticus in cultures from men with prostatitis were more frequent in the third quarter of the year. The bacteria seemed to appear during or after antibiotic treatment but disappeared spontaneously during a follow-up period of six months. Treatment with the surfactant sodium pentosanpolysulphate, a heparinoid, given orally to patients with chronic prostatitis reduced concomitant pain in muscles and joints. The possibility of an altered host factor function in the polymorphonuclear leucocytes of patients with chronic non-bacterial prostatitis colonized with Staph. epidermidis was investigated. Chemotaxis, phagocytosis and intracellular killing were reduced in vitro and may to a part explain the bacteriological findings. Careful evaluations must thus be performed of earlier neglected factors to reach better knowledge of the chronic prostatitis.

Adult↗

Phagocytosis of Staphylococcus aureus by somatic cells in dry cow secretion of mammary gland.

Independently of the stage of the dry period, phagocytosis and killing of Staphylococcus aureus by somatic cells isolated from dry cow secretion were significantly higher in a medium of diluted secretion than in 1% serum and in PBS. Intramammary introduction of vaccine from killed Staphylococcus aureus cells caused, in the steady state dry period, the significant increase of phagocytic and bactericidal activity of somatic cells, examined in the medium of diluted secretion of the mammary gland. The isolated secretion deriving both from vaccinated and non-vaccinated cows causes--in the steady state dry period--the disappearance of the significance of differences between the somatic cells coming from various cows. Phagocytosis is highly significantly correlated with the total and intracellular survival of bacteria; similarly, highly significant correlation exists between the total and intracellular survival of S. aureus.

Animals↗

[Phagocytosis of Staphylococcus aureus and Pseudomonas aeruginosa by pulmonary macrophages and granulocytes].

The elimination (by human alveolar macrophages and pulmonary granulocytes) of one laboratory strain and one wild-type strain each of Staphylococcus aureus and Pseudomonas aeruginosa has been investigated in vitro. Administering 10(6) alveolar macrophages to 5 x 10(6) staphylococci leads to a reduction of viable colony-forming units (CFU) to 5.8 x 10(5) (L] (laboratory strain (L) and 6.9 x 10(5) (wild-type strain (W)) after 60 minutes. The corresponding values of Ps. aeruginosa are 4.3 x 10(6) (L) and 4.4 x 10(6) (W). The peak chemiluminescence values are 9, 160 (L) and 12,300 (W) counts/10(5) cells for Staph. aureus and 7,670 (L) and 8,950 (W) for Ps. aeruginosa. The addition of heat-inactivated serum increases the number of staphylococci observed within alveolar macrophages from 1.57 to 3.18 (L) and from 1.32 to 2.79 (W). Fresh serum enhances ingestion to 8.15 (L) and 9.73 (W) staphylococci/macrophage. The number of phagocytized pseudomonads in pure culture medium is 0.51 (L) and 0.46 (W) bacteria/macrophage. An increase by the addition of serum is not possible. While staphylococci adhere to the membrane of the macrophage over a wide area, the pseudomonads adhere to it in a highly circumscribed manner. The following ingestion steps do not differ from one organism to the other. After 60 minutes, cultivation of 10(6) granulocytes with 5 x 10(6) staphylococci leads to a reduction of the viable CFU to 7.5 x 10(5) (L) and 9.4 x 10(5) (W). In the case of Ps. aeruginosa, the number of surviving CFU is 4.5 x 10(6) (L) and 3.8 x 10(6) (W). After the addition of Staph. aureus, peak values of 37,800 (L) and 35,500 (W) counts/10(5) cells are measured. The corresponding values for Ps. aeruginosa are 28,900 (L) and 33,100 counts/10(5) cells (W). The addition of heat-inactivated serum leads, in the case of Staphylococcus aureus, to an increase in the ingestion of the laboratory strain from 1.07 to 1.94 and of the wild-type strain from 0.94 to 2.41 bacteria/granulocyte. Fresh serum brings about a further increase to 5.52 (L) and 4.82 (W). In the case of Ps. aeruginosa. 0.75 (L) and 0.69 (W) ingested bacteria/granulocyte are observed; the addition of serum fails to bring about any increase. In contrast to the staphylococci, pseudomonads adhere to the granulocyte membrane in a punctate manner only. No differences of ingestion are seen.

Bronchoalveolar Lavage Fluid↗

Methicillin resistant staphylococcus aureus in a teaching hospital of Karachi--a laboratory study.

A search was made for Methicillin Resistant Strains of Staphylococcus Aureus (MRSA) among staph aureus cultures isolated at a teaching hospital in Karachi. Of 100 staphylococcus aureus isolated in 1987-88, 5 were MRSA, four from admitted patients and one from outpatient. These MRSA were resistant to Gentamicin as well as to other antibiotics. The presence of MRSA in the in-patients is a serious problem as it can act as reservoir to cause outbreak of colonisation and infection. No MRSA was isolated from 50 samples studied from Quetta.

Cross Infection↗

Expression of a pseudocapsule by Staphylococcus aureus: influence of cultural conditions and relevance to mastitis.

Strains of Staphylococcus aureus from cases of mastitis in ruminants were cultured in various media and the cells examined electron microscopically for a pseudocapsule. Organisms grown inside the udder, or in nutrient broth supplemented with ovine, bovine or caprine milk whey produced a large, well-defined pseudocapsule outside the cell wall, but such cells had no true capsule when tested by the India ink technique. Modified staphylococcus 110 medium induced secretion by the organisms of copious extracellular slime but did not induce the pseudocapsule. Organisms grown in the presence of milk whey showed a strong propensity to autoagglutinate and produced pseudocapsular material which contained antigens in common with staphylococci grown in vivo. Supplementing media with lactose, dextrose or casein failed to modify the cell surface of S aureus in a manner analogous to that induced by ruminant whey.

Agglutination Tests↗

[The effect of Aerococcus viridans--the basis of the new therapeutic-prophylactic preparation M-bacterin--on the biological properties of Staphylococcus aureus].

Representatives of the normal microflora from genus Aerococcus, in particular strain Aerococcus viridans 167 isolated from breast milk are studied for their effect on biological properties of Staphylococcus aureus in vitro and in vivo. It is established that the number of viable cells of the staphylococcus cultivated in the presence of antagonists in the beef-extract agar decreases progressively with each following passage, the population dying after the seventh-eight passage. Electronograms fix deep changes in the cell ultrastructure. A degree of changes in biological properties depends on the duration of the antagonist action. The results obtained reveal one of the mechanisms of the antagonistic action of aerococci-antagonists producing hydrogen peroxide.

Animals↗

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