Methods for detecting drug-resistant staphylococci in the human body under natural conditions]

Two methods were compared for determination of drug resistant staphylococci on the nasal mucosa of patients, i. e. the routine method for determination of staphylococcal sensitivity to antibiotics and the method of direct plating out of the starting material onto agarized media containing antibiotics. Staphylococci resistant to streptomycin, tetracycline, chloramphenicol, erythromycin, monomycin, axacillin and less frequent to penicillin were found more often with the 2nd method. A method of proportions was developed for testing sensitivity of staphylococci in purulent inflammatory foci of the patients. It provided characterization of the staphylococcal population from the foci by the number of the antibiotic resistant microbial cells.     

Effect of an interferon preparation on increasing bacterial sensitivity to antibiotics

Under the effect of human leukocytic alpha-interferon-II for injections there was observed an increase in sensitivity of bacteria to benzylpenicillin, ristomycin, novobiocin, streptomycin, kanamycin, monomycin and erythromycin. A higher antibacterial effect of the penicillin and erythromycin was recorded only with respect to initially resistant strains of staphylococci. For the antibiotic resistant strains of staphylococci the MICs of the penicillin and erythromycin decreased on the average 17.8 and 208 times (p less than 0.001) respectively. The interferon had no effect on sensitivity of bacteria to chloramphenicol.     

Quantitative Nasal Culture: a Tool in Antibiotic Research

The use of the quantitative nasal culture was investigated as a means of evaluation of new antimicrobial drugs in man. Cyclacillin was somewhat more active in vitro than penicillin G against penicillin G-resistant organisms. Cyclacillin was highly effective in suppressing staphylococci susceptible to penicillin G in nasal carriers but did not suppress staphylococci resistant to penicillin G. Although in previous studies by others cyclacillin was effective in treating mice infected with penicillin G-resistant staphylococci, in the present studies cyclacillin was not effective in suppressing nasal penicillin G-resistant staphylococci in man at doses which markedly suppressed penicillin G-sensitive organisms.     

Trial of Phenoxymethylpenicillin,Phenethicillin, and Lincomycin in Treatment of Staphylococcal Sepsis in a Casualty Department

A comparative trial of phenoxymethylpenicillin (penicillin V), phenethicillin (Broxil), and lincomycin (Lincocin) against superficial staphylococcal infections seen in a casualty department showed no difference in the efficacy of the three agents, though half the staphylococci isolated were resistant to penicillin. Possible reasons include the fact that antibiotic treatment may not affect superficial staphylococcal infections, or that the organisms concerned may have been weak formers of penicillinase.Half the patients treated with lincomycin complained of diarrhoea and 5% of those treated with phenethicillin suffered from nausea.     

Sensitivity to antibacterial preparations of the pathogenic staphylococci isolated from carriers]

After discontinuation of the use of antibiotics for sanation of the carriers of pathogenic staphylococci among the staff of a maternity unit, the number of the strains resistant to penicillin, streptomycin, chlortetracycline, tetracycline, levomycetin, erythromycin, monomycin and neomycin decreased 1.3, 2.9, 10,4.3,5.5,1.5,3.9 and 2.4 times respectively. The differences were statistically reliable. The causes of the antibiotic resistance decrease require further investigation.     

Epidemiological mechanism for the formation of drug-resistant bacterial populations]

The concept on the epidemiological mechanism of formation of drug-resistant bacterial populations was substantiated. Investigations showed the epidemiological mechanism of formation of the drug-resistant populations of staphylococci in the purulent-inflammatory foci to be represented in 81.4 per cent of the patients by superinfection with resistant microbes. The latter group of the patients could be divided into two subgroups: in one of the subgroups (34.9 per cent) a reduction of sensitivity in the initial staphylococci was seen in the presence of donor bacteria which later either remained in the focus or were eliminated. In this case it could be supposed that there occurred a preliminary infection with the resistant staphylococci or with the transducing bacteriophages.     

II. In vitro studies of antibacterial activity

One hundred and forty isolates of beta-hemolytic streptococcus cultured from patients with clinical pharyngitis were studied by disc diffusion for antibiotic sensitivity to lincomycin, erythromycin, cephalexin and penicillin and by agar dilution to cephalexin and penicillin. All isolates were sensitive to ≤ 0.1 μg./ml. penicillin and ≤ 1.56 μg./ml. cephalexin. The disc-diffusion test was reliable in predicting the sensitivities in vitro. One strain of group A betahemolytic streptococcus was resistant to erythromycin by disc diffusion. When compared to Lancefield grouping 18% of strains were incorrectly identified as group A by the bacitracin-disc test. Cephalexin was uniformly effective in vitro in inhibiting beta-hemolytic streptococci and the 30 μg. cephalexin disc was reliable in predicting these sensitivities.     

Comparative characteristics of the antibioticograms and biological properties of the staphylococci isolated from patients with various eye diseases]

A total of 1488 examinations of the conjunctival content of patients with various ophthalmic diseases were performed within 5 years (1973--1977). 989 (66.4 per cent) cultures were isolated. Staphylococci were isolated in 79.2 per cent of the patients irrespective of the diagnosis. The plasma-coagulating strains were isolated in 43.6 per cent of the patients with acute conjunctivitis. The plasma-negative staphylococci were more frequent in chronic conjunctivitis (83.5 per cent). A total of 188 strains were studied according to 18 tests. Determination of the antibiotic sensitivity showed that the number of the benzylpenicillin and tetracycline resistant strains among the plasma-coagulating staphylococci was higher in the patients with acute ophthalmic diseases. In chronic conjunctivitis no differences in the antibiotic sensitivity of the plasma-coagulating and plasma-negative strains were noted. Similar antibiotic sensitivity of all plasma-negative staphylococci irrespective of the diagnosis was found. High biological activity of the plasma-negative staphylococci was shown: 74.1 per cent of the isolates possessed phosphatase, 59.8 per cent possessed protease and 88.4 per cent a lysozyme-like enzyme. The strains isolated in chronic conjunctivitis produced protease more frequently that those isolated in acute ophthalmic diseases.     

Antibiotic sensitivity of the microflora isolated in ENT diseases]

Sensitivity of beta-hemolytic streptococci of group A, Streptococcus viridans, Staphylococcus aureus, epidermal staphylococci, pneumococci, Proteus and Ps. aeruginosa isolated in 1975-1978 from patients with tonsillitis, otitis, sinusitis and other otorhinolaryngological diseases was studied with respect to 19 antibiotics. Data on comparison of the antibiotic sensitivity of the microflora isolated from the patients with otorhinolaryngological diseases in 1964-1974 and 1975-1978 are presented. It was shown that beta-hemolytic streptococci were highly sensitive to all the antibiotics tested except tetracycline. Among Streptococcus viridans the strains resistant to many antibiotics were more frequent than among beta-hemolytic streptococci. Most of the Staphylococcus aureus strains were sensitive to gentamycin, cephaloridin, oxacillin and resistant to the other antibiotics. The epidermal staphylococci were characterized by approximately the same antibiotic sensitivity as Staphylococcus aureus. Resistance of the predominating majority of the Pneumococcus strains to tetracycline was noted. Proteus and Ps. aeruginosa were resistant to all the antibiotics except aminoglycosides. The microflora isolated from the cases with otorhinolaryngological diseases in 1975-1978 were mainly characterized by lower antibiotic sensitivity than that isolated from the cases with the same diseases in 1964-1974. It is possible to suppose that the microorganisms isolated from the patients with otorhinolaringological diseases had no significant differences with respect to their antibiotic sensitivity from those isolated from the patients with other pathological processes.     

Antimicrobial susceptibility of staphylococci isolated from the faeces of wild turkeys (Meleagris gallopavo)

AIMS: The purpose of this study was to investigate the staphylococcal flora associated with wild turkey populations. METHODS AND RESULTS: Faecal samples obtained from 26 wild turkeys over a 16-month period were inoculated onto mannitol salt agar plates to select for staphylococci. Fifty-seven randomly chosen isolates were identified as Staphylococcus lentus and their susceptibility determined against clindamycin, chloramphenicol, ciprofloxacin, erythromycin, oxacillin, penicillin G, rifampin, tetracycline, trimethoprim-sulfamethoxazole, and vancomycin. Resistance was minimal as only 3 isolates showed resistance to clindamycin, 3 isolates were resistant to oxacillin, 3 isolates were resistant to penicillin G, and 1 isolate was resistant to erythromycin. Multiple antibiotic resistance was also minimal. CONCLUSIONS: S. lentus is the predominant staphylococcal species associated with wild turkey faeces and antibiotic resistance in these organisms is not problematic. SIGNIFICANCE AND IMPACT OF THE STUDY: S. lentus has been shown as a potential causative agent of inflammatory reactions in the respiratory tract. Due to increased numbers of wild turkeys and more frequent human exposure, surveys to monitor microbial populations are warranted.     

Relationship between resistance to metallic ions and production of beta-lactamase in strains of Staphylococcus epidermidis.

Thirty five strains of Staphylococcus epidermidis were tested for resistance to penicillin G, erythomycin, metallic ions Zn, As3, As5, Cd, Hg, Pb and activity of beta-lactamase. These studies have shown that the majority of tested staphylococci were resistant to penicillin G, erythromycin, and produced beta-lactamase. No correlation between the activity of beta-lactamase and the resistance to metallic ions has been shown.     

Changes in the ultrastructure of staphylococci in kidney tissue in experimental infection]

A study of ultrathin sections fo the kidney tissue of mice infected with staphylococci showed intracellular localization of the bacteria. In vivo there occurred a morphological reconstruction of external layers of the cell wall of staphylococci, and lysis of individual staphylococci. The middle part of the cell wall and the cytoplasmic membrane proved to be the structures the most stable to lysis. Specific changes of bacteria similar to the changes noted with the action of penicillin on staphylococci in vitro followed effective penicillin therapy of mice infected with staphylococci.     

Bacteriophage Types and Antibiotic Sensitivity of Staphylococci from Bovine Milk and Human Nares

The number, phage types, and antibiotic sensitivity of coagulase-positive staphylococci from grade A raw milk samples produced on 40 farms in the Athens, Ga., milkshed were determined. Counts of mannitol-positive staphylococci in milk ranged from 100 to 3,580 per milliliter, with an arithmetic mean of 1,047. Examination of the nares of 48 dairymen on 34 of the farms also revealed that 13 (27%) were carriers of coagulase-positive staphylococci. Isolates from milk (412) and from nares (39) were tested against the Coles, Seto-Wilson, and International phage sets and 87, 68, and 56%, respectively, proved typable. Nine isolates were not typable. Each of the 33 phages used lysed one or more of the isolates. Staphylococcal phage types per milk sample ranged from 0 to 5, 0 to 7, and 0 to 8, with arithmetic means of 1.9, 2.3, and 2.3, respectively. Of the 13 narial carriers, 7 harbored staphylococci of one or more of the same phage types as those isolated from the milk at the respective farms. Randomly selected isolates were tested against high and low concentrations of 12 common antibiotics. All were either moderately sensitive or resistant to polymixin B. Over 30% were moderately sensitive or resistant to dihydrostreptomycin and penicillin individually. With but few exceptions, all isolates were sensitive to chlortetracycline, bacitracin, carbomycin, chloramphenicol, erythromycin, neomycin, novobiocin, oxytetracycline, and tetracycline individually.     

Antibiotics in management of staphylococcal endocarditis; with special reference to increasing bacterial resistance

Eighteen patients with staphylococcal endocarditis were observed at the Los Angeles County Hospital over a 3-year period (1947-49, inclusive). Twelve died. Bacterial sensitivity studies were carried out in 15 of the cases, and there was resistance to penicillin in ten. Aureomycin was effective in two cases of Staphylococcus aureus endocarditis in which there was no response to penicillin therapy. In one case of Staphylococcus aureus endocarditis the organism was resistant to penicillin and developed increasing resistance to aureomycin.     

Antibiotic sensitivity of staphylococci isolated from the surface of the skin of the breasts in pregnant women

Nine hundred and fourty coagulase-positive and coagulase-negative strains of staphylococci isolated from the skin surface of the mammary glands of 94 pregnant women were tested by the disc agar diffusion method for their sensitivity to five antibiotics. The highest number of the isolates were sensitive to erythromycin and lincomycin (87.7 and 89.7 per cent, respectively). The highest number of moderately resistant strains were detected with respect to methicillin. 19 out of 42 cultures of Staph. aureus were resistant to benzylpenicillin and 24 cultures were resistant to tetracycline. Among staphylococci 130 strains or 13.8 per cent were polyresistant.     

Preferential Inhibition of Penicillinase Induction by Oxytetracycline and Its Effect on the Penicillin Susceptibility of Staphylococci

Exposure of penicillinase-producing staphylococci to a combination of penicillin and oxytetracycline resulted in a synergistic inhibitory activity of the antibiotics on the bacteria. Oxytetracycline was employed in concentrations having little or no effect on bacterial growth. It was found that the synergistic antibacterial effect was caused by the preferential inhibition of penicillinase induction by oxytetracycline, rendering the staphylococci more susceptible to penicillin.     

ANTIBIOTICS IN MANAGEMENT OF STAPHYLOCOCCAL ENDOCARDITIS—With Special Reference to Increasing Bacterial Resistance

Eighteen patients with staphylococcal endocarditis were observed at the Los Angeles County Hospital over a 3-year period (1947-49, inclusive). Twelve died.Bacterial sensitivity studies were carried out in 15 of the cases, and there was resistance to penicillin in ten.Aureomycin was effective in two cases of Staphylococcus aureus endocarditis in which there was no response to penicillin therapy.In one case of Staphylococcus aureus endocarditis the organism was resistant to penicillin and developed increasing resistance to aureomycin.     

Prevention of the formation of drug resistance in bacterial populations by using biologically active substances

The patients with infected wounds of the extremities were treated with kanamycin electrophoresis in combination with chlorhexidine bigluconate, an antiseptic. As compared to the patients treated with kanamycin alone, the rate of the wound size decrease in such patients was 2 times higher. The levels of microbial contamination in these patients were much lower. The contamination level with the aerobic flora was 4.8 times lower, including staphylococci, the level of contamination with which was 5.9 times lower. The contamination level with the kanamycin-resistant bacteria was 22 times lower. The treatment with kanamycin alone resulted in a 2.6-fold increase in the number of the antibiotic-resistant variants in the microbial populations of the wounds. In 48.2 per cent of the patients, this was accompanied by development of resistance to kanamycin in the whole microbial population of the wound. The development of the kanamycin resistance in the staphylococcal populations of 18.1 per cent of the patients was associated with changed sensitivity of the initial strains and in 81.9 per cent of the patients, with superinfection by the resistant strains. No changes in the kanamycin sensitivity of the initial gram-negative organisms during the treatment were observed. The use of chlorhexidine bigluconate, as a biologically active substance in combination with kanamycin potentiated the action of the antibiotic, prevented development and accumulation of the antibiotic-resistant variants in the microbial populations of the wounds and development of the drug resistance in these populations.     

Molecular epidemiology of clinical and carrier strains of methicillin resistant Staphylococcus aureus (MRSA) in the hospital settings of north India

Background

The study was conducted between 2000 and 2003 on 750 human subjects, yielding 850 strains of staphylococci from clinical specimens (575), nasal cultures of hospitalized patients (100) and eye & nasal sources of hospital workers (50 & 125 respectively) in order to determine their epidemiology, acquisition and dissemination of resistance genes.

Methods

Organisms from clinical samples were isolated, cultured and identified as per the standard routine procedures. Susceptibility was measured by the agar diffusion method, as recommended by the Nat ional Committee for Clinical Laboratory Standards (NCCLS). The modified method of Birnboin and Takahashi was used for isolation of plasmids from staphylococci. Pulsed-field gel electrophoresis (PFGE) typing of clinical and carrier Methicillin resistant Staphylococcus aureus (MRSA) strains isolated during our study was performed as described previously.

Results

It was shown that 35.1% of Staphylococcus aureus and 22.5% of coagulase-negative staphylococcal isolates were resistant to methicillin. Highest percentage of MRSA (35.5%) was found in pus specimens (n = 151). The multiple drug resistance of all MRSA (n = 180) and Methicillin resistant Coagulase-negative Staphylococcus aureus (MRCNS) (n = 76) isolates was detected. In case of both methicillin-resistant as well as methicillin-sensitive Saphylococcal isolates zero resistance was found to vancomycin where as highest resistance was found to penicillin G followed by ampicillin. It was shown that the major reservoir of methicillin resistant staphylococci in hospitals are colonized/infected inpatients and colonized hospital workers, with carriers at risk for developing endogenous infection or transmitting infection to health care workers and patients. The results were confirmed by molecular typing using PFGE by SmaI-digestion. It was shown that the resistant markers G and T got transferred from clinical S. aureus (JS-105) to carrier S. aureus (JN-49) and the ciprofloxacin (Cf) and erythromycin (E) resistance seemed to be chromosomal mediated. In one of the experiments, plasmid pJMR1O from Staphylococcus aureus coding for ampicillin (A), gentamicin (G) and amikacin (Ak) resistance was transformed into Escherichia coli. The minimal inhibitory concentrations (MICs) for A and G were lower in E. coli than in S. aureus. However, the MIC for Ak was higher in E. coli transformants than in S. aureus.

Conclusion

There is a progressive increase in MRSA prevalence and multi-drug resistance in staphylococci. Vancomycin is still the drug of choice for MRSA infections. The major reservoir of methicillin resistant staphylococci in hospitals is colonized/infected inpatients and colonized hospital workers. Resistance transfer from staphylococci to E. coli as well as from clinical to carrier staphylococci due to antibiotic stress seemed to be an alarming threat to antimicrobial chemotherapy.     

Characterization of staphylococci isolated from mastitic cows in Spain.

A total of 57 gram-positive, catalase-positive cocci, considered etiological agents of clinical and subclinical bovine mastitis, were tested for glucose and mannitol fermentation, coagulase and thermonuclease production, sensitivity to lysostaphin, gelatin hydrolysis, lysozyme, phosphatase and egg yolk factor production, hemolytic properties, antibiotic sensitivity, susceptibility to human and bovine phages, and enterotoxin production. All 57 strains were identified as staphylococci. A good correlation was found between 3+ and 4+ coagulase reactions, thermonuclease production, and high sensitivity to lysostaphin. Neither mannitol fermentation nor production of other enzymes appeared to be a specific property of bovine Staphylococcus aureus strains. beta- and delta-hemolysins were more frequently found than alpha-hemolysin. Nearly 40% of the strains were penicillin resistant. Strains were lysed by phage 42E from the human phage set more frequently than by phage 42D, whereas with the bovine set, strains were more sensitive to specific bovine phages. Three strains produced enterotoxin C, and one strain produced enterotoxin D.