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W Witte

Publications and source records attributed to W Witte.

At least 19 recordsLinked to original sources

Pulsed-field gel electrophoresis of genomic restriction fragments as a tool for the epidemiological analysis of Staphylococcus aureus and coagulase-negative staphylococci.

Thirteen Staphylococcus aureus and S. epidermidis strains obtained from nose and hand of two employees and one patient of a medical ward as well as two S. hemolyticus strains were analysed according to their restriction fragment length patterns (RFLP) by pulsed-field gel electrophoresis (PFGE) using the restriction enzymes SmaI and SstII. Species identification of the isolates was performed by a system which includes 20 biochemical reactions. Furthermore, the antibiotic resistance patterns of the strains were determined. While several isolates exhibited identical antibiotic susceptibilities and biochemical profiles, differences in the RFLP were obtained. In three cases, S. epidermidis strains colonizing the skin showed an identical restriction profile as isolates from the mucous membranes of the same person. We concluded that the analysis of staphylococcal strains by PFGE is an important epidemiological tool with high discrimination power.

Bacterial Typing Techniques

Occurrence of quinolone resistance in Staphylococcus aureus from nosocomial infection.

Among 63 Staphylococcus aureus isolates (one isolate per one patient) counted from infections (from August to November 1991) in hospital T., eight exhibited resistance to fluoroquinolones. Seven of these quinolone-resistant isolates were multiply- and methicillin-resistant S. aureus (QR-MRSA). The results of phage-, plasmid- and genotyping (pulsed field electrophoresis) revealed that six different strain-clones of these MRSA were spread in the hospital. In vitro spontaneous mutants resistant to fluoroquinolones are 10-100-fold more frequent in MRSA than in other S. aureus when selected on isosensitest-agar containing 1 microgram/ml of ciprofloxacin. However, the same mutant frequencies were found in strain 8325-4 with and without the mecA-determinant. The resistance phenotype was stable over 30 generations of subculture in nutrient broth as well in natural quinolone resistant MRSA as in mutants of other types of S. aureus selected in vitro. The phenotypic association of quinolone resistance and MRSA is rather likely due to a higher frequency of spontaneous resistant mutants which are present in natural populations of MRSA. Data of chemotherapy prior to the isolation of S. aureus show that three of seven patients from whom QR-MRSA were isolated were treated with a quinolone. In eight cases of infections with non-MRSA and quinolone treatment the isolated S. aureus strains were in vitro sensitive to quinolones.

Anti-Infective Agents

Overproduction of a penicillin-binding protein is not the only mechanism of penicillin resistance in Enterococcus faecium.

In 1989 and 1990, a large number of ampicillin-resistant strains of Enterococcus faecium were isolated from infected patients treated at intensive care units in Berlin, Germany. Twenty-five clinical isolates, including five different biotypes as classified by acid production from various sugars and a wide range of susceptibilities to ampicillin (MICs between 0.5 and 128 micrograms/ml), were selected for a detailed analysis of penicillin-binding proteins (PBPs). All strains contained a slowly reacting PBP with low penicillin affinity known to be present in enterococci. Overproduction of this PBP relative to susceptible isolates was noted, especially in all strains for which the MIC of ampicillin was 8 micrograms/ml, to a lesser degree in the more resistant strains, but not at all in the three highly resistant isolates for which the MIC was 128 micrograms/ml. In these three strains, this PBP appears to have a reduced affinity for beta-lactams. The results suggest that overproduction of PBP 6 correlates only with intermediate resistance levels and that higher resistance is mediated by yet another, still unknown mechanism, probably including reduction of beta-lactam affinity in one or more PBPs.

Bacterial Proteins

[Decreasing oxytetracycline resistance in Enterobacteriaceae in veterinary and health conditions after exchange of this substance in animal nutrition].

The national strategy for the use of chemotherapeutics under which there is a distinction between therapeutics and growth promoters for all uses (human medicine, veterinary medicine, animal nutrition) has resulted in a distinct reduction of the selection pressure and a decrease of the OTC resistance among Enterobacteriaceae in the eastern German Länder (formerly GDR) due to a reduction of OTC feeding after 1981 and the final prohibition in 1983. In the following years Enterobacteriaceae were investigated as infectious pathogens for humans and animals using antibiograms. On the one hand, the OTC resistance of porcine and bovine E. coli strains decreased only by 27% and 17%, respectively, during the 1980 to 1988 period, on the other, the OTC resistance of bovine S. typhimurium dropped by 65%, porcine S. typhimurium by 50%, bovine S. dublin by 50%, porcine S. choleraesuis by 30% to a resistance level of about 13% (1989) demonstrating a reduced pressure of resistance. At the same time, OTC resistance became diminished from 50% and 70%, respectively, to 30% also in Enterobacteriaceae (E. coli, Klebsiella) and enterococci isolated from urine and wound infections of patients in the non-agricultural population although therapeutic habits had remained unchanged. The results shown point to a spreading of resistance plasmids which has to be expected far beyond the field of OTC use, for example after the use of antibacterial growth promoters in livestock farming. Control measures should be introduced.

Animal Feed

[Differentiation of methicillin-resistant strains of Staphylococcus aureus by prophage specificity].

By inducing with mitomycin C the following phages were isolated from all the tested 32 methicillin resistant strains of S. aureus: the serogroup B phage was isolated from 2 strains, the serogroup B and F phages were isolated from 5 strains and the serogroup F phage was isolated from 25 strains. The phages were divided into 5 groups by the antiphage immunity. In group 1 of the phages 4 additional phages were specified. By the specificity of the prophages in the cultures all the strains were divided into 5 groups. Group 1 of the cultures was divided into 5 subgroups (A, B, C, D and E).

Bacteriophage Typing

[Effect of prophages on transfer frequency of conjugative plasmid G 873].

Transfer of the conjugative plasmid G873 on filters and mixed cultivation of the donor and recipient cells in liquid media is described. In the both systems the use of the lysogenic recipient cells (phages of serogroups B and F) in the crossings increased mor than 100-fold the frequency of plasmid transfer. The conjugative transfer of the plasmid in the mixed cultivation system was proved. The conjugative transfer required the presence (while not obligatory) of calcium chloride and was restricted by the serum factors.

Animals

Instability of in vitro resistance to imipenem in Pseudomonas aeruginosa.

In 3 from 19 clinical isolates of Pseudomonas aeruginosa imipenem-resistant subpopulations could be detected in vitro. In comparison to their wild strains these imipenem-resistant cells were lacking an outer-membrane-protein of 50 kD. In the subpopulation derived from Pseudomonas aeruginosa 76 resistance to imipenem was found to be instable. It could be lost within a short period of time after subcultivation in the absence of imipenem. Cells with restored sensitivity to imipenem possess the outer membrane protein of 50 kD as the wild strain does.

Azlocillin

Typing of multiple isolates of coagulase-negative staphylococci from blood cultures and cerebrospinal fluid in neonates and children: an approach to discriminate contaminants.

From eight neonates and children suspicious for suffering from septicemia and/or meningitis, coagulase negative staphylococci (CNS) were isolated from multiple blood cultures and from cerebrospinal fluids originating from different punctures. S. epidermidis was the predominant species. By a further typing using plasmid-profiles and exoprotein patterns besides the resistance phenotype, contaminants could be discriminated from the isolates as the probable etiologic agent exhibiting unique reaction patterns. CNS isolated from cases of septicemia together with other bacterial pathogens or from cases of presumed but not confirmed septicemia revealed as different.

Adolescent

A single clone of Staphylococcus aureus causes the majority of cases of toxic shock syndrome.

Genetic relationships among 315 isolates of the bacterium Staphylococcus aureus expressing toxic shock syndrome toxin-1 (TSST-1) recovered primarily from humans with toxic shock syndrome (TSS) in five countries on two continents were determined by analyzing electrophoretically demonstrable allelic variation at 20 chromosomal enzyme loci. Forty-nine distinctive electrophoretic types (ETs), representing multilocus enzyme genotypes, were identified. Cluster analysis of the ETs revealed two major phylogenetic divisions separated at a genetic distance of 0.35 and seven branches diverging from one another at distances greater than or equal to 0.20. A single clone (ET 41) accounted for 88% of cases of TSS with a female urogenital focus and 53% of TSS cases involving nonurogenital (predominantly wound) infections. With few exceptions, strains representing different phylogenetic lines had characteristic TSST-1 gene (tst) restriction fragment length polymorphism patterns obtained by digestion of genomic DNA with Cla I. Strains recovered from ovine and bovine hosts with mastitis were genotypically distinct from the major human TSS clone. The expression of TSST-1 in cell lineages representing the total breadth of multilocus genotypic diversity in the species S. aureus as a whole is interpreted as evidence that the TSST-1 gene is evolutionarily old. The recovery of a single clone from the majority of individuals afflicted with TSS having a urogenital focus and from the genital tract of a large proportion of asymptomatic female carriers strongly suggests that this clone is especially well adapted for colonization of these anatomic sites.

Animals

Activity of clavulanate-potentiated penicillins against methicillin-resistant Staphylococcus aureus.

It has been suggested that combinations of penicillins with clavulanate may be useful in treatment of infections by methicillin-resistant strains of Staphylococcus aureus (MRSA). To determine the potentiating effect of clavulanate on the antibacterial activity of penicillins, we studied MRSA in vitro by an agar-dilution method. A total of 124 clinical isolates of MRSA were tested for sensitivity to benzylpenicillin, amoxycillin, and ticarcillin alone and in combination with 1.25, 2.5, 5.0 and 10.0 mg/l of clavulanate. Most of the strains were not typable by the international reference set of bacteriophages of human staphylococci but showed typical properties of nosocomial strains. A reduction in the MIC90 of benzylpenicillin and amoxycillin to 25 mg/l was seen in the presence of 2.5 mg/l of clavulanate. The effect was less pronounced with ticarcillin. In spite of some increase in the susceptibility of MRSA to benzylpenicillin and amoxycillin produced by clavulanate, these combinations seem to be inappropriate in infections due to MRSA.

Clavulanic Acids

[Nosocomial infections caused by multi-resistant Serratia marcescens at a university clinic--clinical aspects and drug resistance].

Serratia marcescens (S.m.) has become increasingly important as a nosocomial pathogen and displayed an increasing resistance to antimicrobial agents in the past decade. We recently studied in 1985 and 1986 an epidemic caused by multi resistant S.m. strains that involved 27 infants and 1 adult patient. 14 neonates (in most cases very low birth weight infants) in a neonatal intensive care unit developed a S.m.-septicemia and/or meningitis, 11 of them died. In a ward for young infants with congenital heart diseases 13 patients suffered a S.m. infection and one patient died in the adult intensive care unit in consequence of a S.m. septicemia.

Anti-Bacterial Agents

Complex typing of methicillin-resistant Staphylococcus aureus (MRSA).

To discriminate between methicillin-resistant Staphylococcus aureus from 5 nosocomial outbreaks and from sporadic nosocomial infections, the efficacy of a complex typing scheme by phage typing, biochemical typing, resistance phenotype, plasmid profiles, plasmid patterns and attribution of resistance determinants to the chromosome was studied. In addition to the International Basic Set and experimental phages 88-93, 10 experimental phages from the Public Health Laboratory Service, Colindale, London, were used for phage-typing. The 10 experimental phages from PHLS in particular, in combination with plasmid profiles and plasmid patterns, were of special discriminative value.

Bacterial Typing Techniques

[Coagulase-negative Staphylococcus species in mixed fodder and on grain].

For valuation of microbially-altered combined fodder from the veterinary point of view beside the total number of germs also the diagnosis of the species of veterinary relevant indicator germs is of importance. Coagulase-negative staphylococci are the main and indicator flora in spoiling of combined fodder. According to the biochemical and physiological characterization the majority of the coagulase-negative staphylococci occurring in connection with spoiling of combined fodder belongs to the species S. xylosus. Beside the species also other coagulase-negative species as S. sciuri, S. cohnii, S, warneri and S. saprophyticus have been demonstrated. These bacterial species were also found in the single components of combined fodder as ripe grain, green unripe cereal plants, in soil samples and in the dust inside the mills for combined fodder production.

Animal Feed

Staphylococcus aureus strains of the 94/96 complex isolated in the German Democratic Republic: incidence and discrimination of strain clones.

The incidence of S. aureus with a phage pattern of 94/96 rose from 9% in 1979 to 18% in 1985. The frequency of occurrence not only increased among isolates from inpatients and outpatients but also among those from healthy carriers. All of the 504 investigated strains of different origin exhibited a uniform pattern of biochemical characteristics. In each of 40 investigated strains, a plasmid with a molecular mass of 16 Md was found. Elimination experiments indicated that these plasmids determined resistance to penicillin and/or cadmium. Resistance to chloramphenicol was found to be determined by plasmids of 2.0 Md, resistance to oxytetracycline by plasmids of 2.7 Md. Clones could be discriminated by means of 7 experimental phages. The application of these phages for typing strains from infections in hospitals is demonstrated.

Bacteriophage Typing

Multiply- and methicillin-resistant Staphylococcus aureus strains isolated in the German Democratic Republic in 1985 and 1986.

Multiply- and methicillin-resistant Staphylococcus aureus (MRSA) strains have been isolated from five small outbreaks of nosocomial infection in five different hospitals. The MRSA were typed by phage patterns, biochemical traits, resistance phenotypes and plasmid patterns. Three different groups of strains can be distinguished. The MRSA from three outbreaks in one country share identical characters. Phage typing by the use of the International Basic Set for Phage Typing staphylococci as well as experimental phages does not completely discriminate between the strains. Attribution of several resistance determinants to plasmids in two of the described strain groups proved valuable for strain differentiation. These multiply-resistant strains are sensitive to vancomycin and to rifampicin.

Cross Infection

Spread of plasmid-mediated nourseothricin resistance due to antibiotic use in animal husbandry.

After using of the streptothricin antibiotic nourseothricin in animal husbandry for growth promotion, plasmid-borne resistance to streptothricin could be observed in E. coli from nourseothricin fed pigs, from employees in pig farms and from their family members. Moreover, streptothricin resistance plasmids also occurred in E. coli of man without any contact to pig farms (gut flora and even urinary tract infections). However, these individuals live in villages and towns of the territory where nourseothricin was applied to pigs. Similar streptothricin resistance plasmids belonging to different incompatibility groups were found in both E. coli from pigs and E. coli from human beings. As no coselection of resistance to drugs indispensable for therapeutic use in man was observed, the application of nourseothricin in animal husbandry has not clinical implication for human medicine yet. Nevertheless, this problem remains under further investigation.

Animal Husbandry