Clonal expansion of Staphylococcus epidermidis strains causing Hickman catheter-related infections in a hemato-oncologic department

The detailed analysis of 411 strains of coagulase-negative staphylococci (CoNS) obtained from 40 neutropenic hemato-oncologic patients (61 Hickman catheter episodes) on intensive chemotherapy is described. By random amplification of polymorphic DNA (RAPD) analysis, a total of 88 different genotypes were detected: 51 in air samples and 30 in skin cultures prior to insertion, 12 in blood cultures after insertion, and only 5 involved in catheter-related infections (CRI). Two RAPD genotypes of Staphylococcus epidermidis predominated, and their prevalence increased during patient hospitalization. At insertion, these clones constituted 11 of 86 (13%) CoNS isolated from air samples and 33 of 75 (44%) CoNS isolated from skin cultures. After insertion, their combined prevalence increased to 33 of 62 (53%) in catheters not associated with CRI and 139 of 188 (74%) in catheters associated with CRI (P = 0.0041). These two predominant S. epidermidis clones gave rise to a very high incidence of CRI (6.0 per 1,000 catheter days) and a very high catheter removal rate for CRI, 70%, despite prompt treatment with vancomycin. A likely source of S. epidermidis strains involved in CRI appeared to be the skin flora in 75% of cases. The validity of these observations was confirmed by pulsed-field gel electrophoresis (PFGE) of SmaI DNA macrorestriction fragments of blood culture CoNS isolates. Again, two predominant CoNS genotypes were found (combined prevalence, 60%). RAPD and PFGE yielded concordant results in 75% of cases. Retrospectively, the same two predominant CoNS clones were also found among blood culture CoNS isolates from the same hematology department in the period 1991 to 1993 (combined prevalence, 42%) but not in the period 1978 to 1982. These observations underscore the pathogenic potential of clonal CoNS types that have successfully and persistently colonized patients in this hemato-oncology department.     

Aminoglycoside resistance patterns of Serratia marcescens strains of clinical origin

Aminoglycoside resistance patterns of 147 Serratia marcescens strains of clinical origin were studied. All strains analysed belonged to three different bacterial populations. The periods of study and the institutions the strains were isolated from correlated significantly with the resistance patterns shown by the strains. The most frequent resistance patterns found were the following: ACC (6')-I at the Hospital Infantil de México (Children's Hospital of México), and ANT (2') + AAC(6')-I at the Instituto Nacional de Pediatría (INPed or National Institute of Pediatrics) in Mexico City. Furthermore, the isolation frequency of aminoglycoside-sensitive strains decreased remarkably at the INPed over a 12-year period. These results suggest that there has been a selection of Serratia marcescens strains that are very resistant to aminoglycosides.     

Molecular analysis of and identification of antibiotic resistance genes in clinical isolates of Salmonella typhi from India

A representative sample of 21 Salmonella typhi strains isolated from cultures of blood from patients at the Christian Medical College and Hospital, Vellore, India, were tested for their susceptibilities to various antimicrobial agents. Eleven of the S. typhi strains possessed resistance to chloramphenicol (256 mg/liter), trimethoprim (64 mg/liter), and amoxicillin (>128 mg/liter), while four of the isolates were resistant to each of these agents except for amoxicillin. Six of the isolates were completely sensitive to all of the antimicrobial agents tested. All the S. typhi isolates were susceptible to cephalosporin agents, gentamicin, amoxicillin plus clavulanic acid, and imipenem. The antibiotic resistance determinants in each S. typhi isolate were encoded by one of four plasmid types. Plasmid-mediated antibiotic resistance genes were identified with specific probes in hybridization experiments; the genes responsible for chloramphenicol, trimethoprim, and ampicillin resistance were chloramphenicol acetyltransferase type I, dihydrofolate reductase type VII, and TEM-1 beta-lactamase, respectively. Pulsed-field gel electrophoresis analysis of XbaI-generated genomic restriction fragments identified a single distinct profile (18 DNA fragments) for all of the resistant isolates. In comparison, six profiles, different from each other and from the resistance profile, were recognized among the sensitive isolates. It appears that a single strain containing a plasmid conferring multidrug-resistance has emerged within the S. typhi bacterial population in Vellore and has been able to adapt to and survive the challenge of antibiotics as they are introduced into clinical medicine.     

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This study is a retrospective review of admissions, discharge records and blood culture results of neonates admitted to the Neonatal Intensive Care Unit of Korle Bu Teaching Hospital in Accra, from the first of January 1991 to the 31st of December 1992. During this two year period there were 443 positive blood cultures. Ninety percent of the blood cultures were from babies born in Korle Bu Teaching Hospital, thus making the incidence of neonatal bacteraemia 22.2 per 1000 live births. The overall mortality rate was 37.2%. Gram negative bacteria accounted for 70.9% and Gram positive bacteria for 29.1% of all neonatal bacteraemia. The most common isolates were Enterobacter species 29.6%; Streptococcus faecalis 14.4%; Staphylococcus aureus 10.8%; Acinetobacter species 9.5%; Klebsiella species 9% and Escherichia coli 8.8%. It is concluded that the incidence of neonatal bacterial sepsis is high in our hospital and is associated with a very high mortality rate. There is thus an urgent need to institute appropriate preventive and therapeutic measures.     

Rapid diagnosis of cholera caused by Vibrio cholerae O139

Hybridomas secreting specific monoclonal antibodies (MAbs) to Vibrio cholerae serogroup O139 were produced. Six monoclones (hybridomas) secreting MAbs specific only to lipopolysaccharide of V. cholerae O139 strains and which did not cross-react to 137 strains of other enteric microorganisms were obtained. These clones were designated 12F5-G11, 12F5-G2, 15F5-H5, 5B9-F8, 14C9-D2, and 6D2-D8. The immunoglobulin (Ig) heavy chain isotypes secreted by these clones were IgG2b, IgG2b, IgG2b, IgM, IgG2b, and IgG3, respectively. Clone 12F5-G11 was selected for mass production of MAb, which was used as a detection reagent in the antigen detection assay for diagnosis of cholera caused by V. cholerae O139, and this assay was compared to the conventional bacterial isolation method. Five batches of rectal swab cultures in alkaline-peptone water were collected from 6,497 patients with watery diarrhea. These were 6,310 patients admitted to Bamrasnaradura Infectious Diseases Hospital, 16 patients from Krung Thon Hospital, 78 patients from Bangkok Children's Hospital, 19 patients from Karen refugee camps, and 74 Indian patients from the National Institute of Cholera and Enteric Diseases, Calcutta, India. The V. cholerae O139 isolations from the rectal swab cultures and the antigen detection assays (i.e., the MAb-based dot-blot ELISA) were performed by different persons of different laboratories, and the results were revealed after all specimens had been tested. Of the 6,497 samples tested, the dot-blot ELISA correctly identified 42 of 42 V. cholerae O139-positive samples and gave a result of positive for three samples which were culture negative for V. cholerae O139. The diagnostic sensitivity, specificity, and efficacy of the dot-blot ELISA were 100, 99.95, and 99.26%, respectively. The ELISA is easy to perform and relatively inexpensive. It can test multiple samples at a single time, does not require special equipment, and does not produce great quantities of contaminated waste. Most of all, it reduces the diagnostic time from at least 2 days for the bacterial isolation to less than 90 min. The assay is recommended as a rapid screening test of cholera cases caused by V. cholerae O139.     

Toxoplasmosis infection in pregnant women in Lanzhou

A strain of poliovirus type 1 isolated from an endemic in Shandong Province in 1988 and three strains of poliovirus type 1 isolated from an endemic in Xinjiang in 1990 were analysed by dot blot hybridization with Sabin I specific probe and PCR amplification with Sabin I specific primers. They were proved to be non-Sabin-like poliovirus. The nucleotide sequences of VP1-2A junction region of four poliovirus type 1 isolates were analysed. It was found that the nucleotide diversity of four isolates with Sabin I was equal to or greater than 16.7%. This finding proved that they were wild polioviruses and greatly different fromSabin I. It was also found that the nucleotide sequence diversity among three strains from Xinjiang was very small (less than 2.6%) with identical amino acid sequences. This indicated that the three Xinjiang strains belonged to the same genotype. While the difference of Shandong strain from the three Xinjiang strains was between 14. 0% to 15.33% in nucleotide sequence and 2.0% to 4.0% in amino acids. This indicated that Shandong strain was apparently different from three Xinjiang strains and it belonged to another wild genotype which had a different history of evolution.     

Blood cultures positive for coagulase-negative staphylococci: antisepsis, pseudobacteremia, and therapy of patients

A blood culture cohort study investigating issues related to isolation of coagulase-negative staphylococci (CoNS) and other skin microflora is reported. Data were collected over 12 weeks to determine the incidence of significant CoNS bacteremia versus that of pseudobacteremia (contaminants) and to evaluate drug therapy in patients with cultures positive for CoNS. In addition, the effectiveness of 0.2% chlorine peroxide as a bactericidal disinfectant was compared to that of 10% providone iodine. A total of 3,276 cultures of blood from 1,433 patients were evaluated in the study. Eighty-nine cultures were positive for skin flora, with 81 of 89 (91%) involving CoNS. The incidence of significant CoNS bacteremia was 20 of 81 (24.7%), that of indeterminate bacteremia was 10 of 81 (12.3%), and that of contamination was 59 of 81 (72.8%). The incidence of significant bacteremia involving CoNS was double the 10 to 12% rate based on previous estimations at our institutions. In tests with the two bactericidal disinfectants, 22 of 1,639 cultures (1.3%) in the chlorine peroxide group versus 37 of 1,637 (2.3%) in the providone iodine group were considered contaminated (P = 0.065). Rates of contamination for venipuncture versus catheter collection were not significantly different (P = 0.46). The overall contamination rate was 59 of 3,276 (1.8%), which is consistent with the lower end of published quality assurance benchmark standards. The low rate was believed to be due to the professional phlebotomy staff in our institutions. There was excellent agreement between retrospective analysis by reviewers, when formal criteria were used, and the attending physicians' intuitive clinical impressions in the classification of significant bloodstream infections (100% agreement) or contamination (95% agreement). However, physicians still used antimicrobial agents to treat nearly one-half of the patients with contaminated blood cultures, with vancomycin being misused in 34% of patients. In addition, 10% of patients with significant bacteremia were treated with inappropriate agents. There were no significant adverse events or prolonged hospital stays due to the unnecessary use of vancomycin; however, the additional costs of treating patients whose cultures contained CoNS contaminants was estimated to be $1,000 per patient. Measures to limit the unnecessary use of vancomycin (and other agents) are important.     

Biovars and antibiotic resistance of enterobacter cloacae strains isolated from colonized newborns

A total of 120 strains of E. cloacae were isolated from colonized newborns in a maternity hospital (96 strains) and from inpatients of other hospitals (23 strains). Biovars of these strains and of 1 reference strain were determined by two methods of biochemical typing and their sensitivity to 14 antibiotics assessed. The overwhelming majority of isolates from newborns were referred to type 2 according to typing after Old (original) or to biovar 62 if typed using the modification of this method. The phenotypical profile of determinants of resistance to 10 antibacterial drugs were the same, including those to aminoglycosides (except amikacin) and third-generation cephalosporins. The strains isolated from non-obstetrical inpatients belonged to biovars with phenotypical profiles of resistance to only 3 antibiotics and sensitive to aminoglycosides and third-generation cephalosporins.     

Molecular characterization of penicillin-resistant Streptococcus pneumoniae isolates from Bulgaria

As part of an ongoing surveillance program of antibiotic-resistant Streptococcus pneumoniae in Sofia, Bulgaria, 120 penicillin-resistant strains (PRSP) (most of them recovered from children hospitalized with pneumococcal disease) were analyzed by microbiological and molecular methods. Several unique features of this collection are of particular interest. (i) Most isolates (112 of 120) were also resistant to trimethoprim-sulfamethoxazole (SXT) (97 of 120 isolates, or 80%), and over 70% (86 of 120) of the isolates were resistant to at least three antibiotics in addition to penicillin. (ii) Close to 80% of all isolates were represented by large clusters of bacteria, each with a unique serotype, antibiotype, and chromosomal macrorestriction pattern (determined by pulsed-field gel electrophoresis), as well as unique restriction fragmentation length polymorphisms of the penicillin-binding protein genes pbp1a, pbp2x, and pbp2b. (iii) A large proportion (45 of 120, or 38%) of the strains belonged to two internationally spread epidemic clones of S. pneumoniae, the first expressing capsular type 23F and the second expressing serotype 9. (iv) A unique Bulgarian cluster composed of eight serotype 19F isolates was resistant to tetracycline, SXT, cefotaxime, and extremely high levels of penicillin and erythromycin. Nevertheless, this clone did not react with either the erm or the mef DNA probes, and thus the mechanism of macrolide resistance in this group of PRSP remains to be elucidated.     

Study of the antibiotic sensitivity of Shigella newcastle isolated in Zaporozhe Province

Sensitivity to antibacterial drugs of 510 strains of Shigella Newcastle isolated from both sporadic patients and cases during the disease outbreaks was studied. The studies revealed a rather high percentage of Shigella Newcastle resistant to the antibiotics widely used in clinical practice. Most of the strains, i.e. 90.5 per cent were resistant to tetracycline and a significant number of the isolates, i.e. 55.6--53.3 per cent were resistant to streptomycin and erythromycin. A statistically reliable predominance of the Shigella resistant to levomycetin and streptomycin in the disease foci was noted as compared to the Shigella isolated from the sporadic patients. Most of the isolates were simultaneously resistant to several antibiotics. Thus, resistance to 2,3--4 and 5--6 antibiotics was found in 28, 54.3 and I.I per cent of the cultures respectively. At the same time almost all Shigella Newcastle strains were sensitive to rifampicin, neomycin and monomycin (aminoglycoside antibiotics), enteroseptol (oxychinoline drug) and furazolidone (nitrofuran drug).     

Bile duct bacterial isolates in primary sclerosing cholangitis: a study of explanted livers

BACKGROUND/AIMS: The pathogenesis of the inflammatory lesion in primary sclerosing cholangitis is unknown. The clinical picture is characterized by i.a. episodes of fever, the cause of which also remains speculative. Previous studies of bacterial isolates in the liver or bile ducts in primary sclerosing cholangitis have had the shortcoming of possible contamination associated with the sampling. The aim of this study was to investigate whether bile and bile duct tissue, obtained under sterile conditions in connection with liver transplantation, contain bacteria. METHODS: We studied bile from bile duct walls and bile collected from the explanted livers of 36 patients with primary sclerosing cholangitis and 14 patients with primary biliary cirrhosis. RESULTS: Positive cultures were obtained from 21 of 36 primary sclerosing cholangitis patients, but from none of the primary biliary cirrhosis patients. The number of bacterial strains was inversely related to the time after the last endoscopic retrograde cholangiography. Treatment with antibiotics or intraductal stent, or the occurrence of fever before liver transplantation did not seem to influence the culture results, whereas antibiotic treatment in connection with endoscopic retrograde cholangiography may possibly have reduced the number of isolates in the cultures. Alpha-haemolytic Streptococci were retrieved as late as 4 years after the last endoscopic retrograde cholangiography. Retrospective analysis of liver laboratory tests after endoscopic retrograde cholangiography did not indicate a deleterious effect of the investigation. CONCLUSIONS: The data suggest that antibiotics should be given routinely in connection with endoscopic retrograde cholangiography. They also raise the question of a possible role of alpha-haemolytic Streptococci in the progression of primary sclerosing cholangitis.     

Surveillance of antibiotic resistance in Neisseria gonorrhoeae in The Netherlands, 1977-95

OBJECTIVE: To evaluate the prevalence and epidemiology of penicillinase producing Neisseria gonorrhoeae (PPNG) and tetracycline resistant N gonorrhoeae (TRNG) in the period 1977-95 in the Netherlands. To compare auxotypes, serovars, and antibiograms of PPNG, non-PPNG, and TRNG. To identify determinants in patient characteristics for the epidemic spread of TRNG/PPNG. METHODS: With respect to the national gonococcal surveillance all PPNG isolates from 30 laboratories over the country in 1977-90 and all gonococcal isolates from five sentinel laboratories (during 1 month per quarter) in 1991-5 were collected. Isolates were auxotyped and serotyped, the susceptibility for various antibiotics was tested and plasmid contents were evaluated. Additional data on PPNG infected individuals were collected retrospectively during a microepidemic of TRNG/PPNG. Univariate and multivariate analyses were performed to identify risk factors for TRNG/PPNG infections. RESULTS: In 1995 an overall high prevalence of PPNG infection (27%) and TRNG among PPNG infection (24%) was found in the Netherlands. Importantly, PPNG were found to have higher MICs for ceftriaxone and ciprofloxacin than non-PPNG; clinically relevant resistance to these antibiotics (or related agents) may emerge first among these strains. The observed diversity of strains (123 auxo/serovar classes since 1988) indicates a continuous introduction of new strains into the community. The epidemic increase of TRNG/PPNG was mainly caused by A/S classes NR/1B-6, PRO/1A-3, and PRO/1A-6, suggesting a clonal spread of a few strains; the rapid spread was associated with transmission in high risk individuals (that is, prostitutes and their clients). CONCLUSION: The prevalence of PPNG in the Netherlands remains high and reduced sensitivity to other antimicrobials was detected among the PPNG strains. This underlines the necessity for a continuous national surveillance of resistance in gonococci including limited epidemiological information.     

Ventriculoperitoneal shunt infections with gram-negative bacteria

Infection causes major morbidity and mortality in patients with cerebrospinal fluid (CSF) shunts. The prognosis of CSF shunt infections caused by Gram-negative bacteria (GNB) has been thought to be particularly poor. The authors reviewed all GNB shunt infections treated at Children's Memorial Hospital from January 1986 to January 1990 (n = 23). Of these infections 20 (87%) occurred within 4 weeks after shunt revision (median, 10 days). The most frequent symptoms were fever, lethargy, and irritability; the illness was not severe in the majority of these patients. Escherichia coli was isolated from 12 of 23 patients (52%), Klebsiella pneumoniae from 5 (22%), and mixed GNB from 3 (13%) patients. Initial treatment always included immediate shunt removal, externalized ventricular drainage, and intravenous antibiotics. Extraventricular drainage revision and/or intraventricular antibiotics were required in four patients whose CSF cultures were persistently positive for GNB. At admission, these patients had CSF glucose levels of < 10 mg/dl and CSF positive for GNB by Gram's stain. The overall cure rate was 100%, and no recurrence was observed; however, a subsequent infection with a different organism developed in four patients. Only 2 of 19 patients (11%) who were followed up suffered apparent CNS damage. One patient died of unrelated causes shortly after treatment. Our findings indicate that 1) patients with GNB CSF shunt infections often appear relatively well at presentation; 2) CSF positive for GNB by Gram's stain and very low CSF glucose levels predict continued positive CSF cultures, despite appropriate antibiotic therapy; and 3) GNB CSF shunt infections can be successfully treated by prompt shunt removal, extraventricular drainage, and intravenous antibiotics.(ABSTRACT TRUNCATED AT 250 WORDS)     

Toxic shock syndrome in 8 children

OBJECTIVE: Evaluation of patients with toxic shock syndrome (TSS) in a paediatric hospital. DESIGN: Retrospective analysis. SETTING: Paediatric Intensive Care Unit, University Hospital, Rotterdam, the Netherlands. METHOD: Analysis of the medical records on 155 patients admitted between January 1982 and January 1992 suffering from shock, 8 of whom had TSS. RESULTS: Five out of 8 TSS patients were under 5 years of age. All the patients needed mechanical ventilation. All patients survived. In 7 patients a probably causative focus of infection was found. The cultures of 6 patients showed growth of Staphylococcus aureus, those of 2 patients showed Lancefield group A beta-haemolytic streptococci (bacterial culture in one, increased antibody titer in the other). Systematic phage typing was not performed. CONCLUSION: Although TSS is a relatively rare disease in young children, it is a potentially lethal one, early recognition of which is very important.     

Development and evaluation of a selective and indicative medium for isolation of Actinobacillus pleuropneumoniae from tonsils

In order to isolate Actinobacillus pleuropneumoniae from mixed bacterial flora a selective and indicative medium was developed. The optimal concentrations of antibiotics were determined for selective chocolate agar (S-TSA) and selective blood agar (S-MBA) using a set of 25 strains of A. pleuropneumoniae. Following optimization of the media 96 isolates of A. pleuropneumoniae were tested on S-TSA and S-MBA. For isolation of A. pleuropneumoniae from 101 pig tonsils collected at slaughter the S-MBA proved significantly better than both S-TSA and the two non-selective agars tested. Furthermore, contaminating to S-TSA and the two non-selective media.     

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Over a one-year period, all coagulase-negative staphylococci (CoNS) from blood cultures, cerebrospinal fluids and peritoneal effluents from patients in a major Danish university hospital were investigated for susceptibility to penicillin G; methicillin; gentamicin; netilmicin; amikacin; erythromycin; clindamycin; fusidic acid; rifampicin; tetracycline; chloramphenicol; ciprofloxacin; teicoplanin; and vancomycin. Among the CoNS-isolates, 56% were resistant to methicillin, 51% to gentamicin, 28% to ciprofloxacin, and 5% to teicoplanin. Blood culture CoNS-isolates from patients with a central venous catheter (CVC) were more often resistant to various antibiotics compared to CoNS-isolates from patients without a CVC, e.g. methicillin (72% vs 21%), gentamicin (65% vs 22%) (p<0.00000001). Great diversity in antibiotic resistance between the wards was found; methicillin resistance (in most cases multiple antibiotic resistance) was in particular associated with consumption of broad-spectrum beta-lactams, quinolones, and total antibiotic consumption in a ward. Thus, the antibiotic policy of a ward is an important factor for antibiotic resistance among CoNS.     

Bacterial pathogens and their antimicrobial susceptibility in Kumasi, Ghana

Between January, 1994 and June, 1996 a survey of bacterial isolates from clinical specimens and their antimicrobial susceptibility was performed at the Komfo Anokye Teaching Hospital, Microbiology Department, Kumasi, Ghana. A total of 11,380 bacterial isolates were cultured from eight different specimens. The sites of origin were wounds 32.2%, urine 28.1%, ear, nose and throat 3.6%, sputum 2.5% and aspirates 2.5%. Gram-negative bacteria accounted for 7955 (69.9%) isolates, the main species were Escherichia coli 47.1%, Pseudomonas spp. 16.8%, Proteus spp 14.6%, Klebsiella spp 10.2%, Neisseria gonorrhoeae 4.2%, Gram-positive bacteria contributed 3425 ((30.1%) of isolates, with Staphylococcus aureus 54.6% being the most predominant followed by Coagulase negative Staphylococcus 18.1%, Streptococcus pneumoniae 13.7% and Beta-haemolytic streptococci 4.1%. Escherichia coli showed 88% and 82% resistance to ampicillin and cotrimoxazole respectively with 78% being susceptible to gentamicin. Cefuroxime resistance in Gram-negative bacilli was 5%. As much as 30.6% and 21.7% of Streptococcus pneumoniae isolates were resistant to Penicillin and chloramphenicol respectively. Ten per cent of Staphylococcus aureus strains were susceptible to penicillin and 18% were resistant to flucloxacillin.     

Allelic polymorphism and site-specific recombination in the opc locus of Neisseria meningitidis

The opc gene is widespread in epidemic and endemic Neisseria meningitidis, but most strains of certain epidemic clones (ET-37 complex, Cluster A4) and a few random endemic isolates lack an opc gene. Four percent of the 1148 bp that contain opc plus the surrounding intergenic region was polymorphic (18 alleles), and many of the alleles contained a 230 bp insertion at a fixed location in the intergenic region. The presence or absence of the insertion reflects site-specific recombination. The alleles are stably inherited within clonal groupings for up to at least 50 years, with rare cases of horizontal genetic exchange. Most statistical methods indicated significant intragenic recombination events within this dataset.     

Phage types and ribotypes of Salmonella enteritidis in southern Italy

Differently from other European countries, Southern Italy was affected by a considerable increase in human infections due to Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) only after 1990. On the present investigation, two groups of S. Enteritidis strains isolated during the low-incidence period 1980-1984 and the epidemic period 1990-1993, respectively, have been submitted to phage-typing and ribotyping in order to ascertain whether the epidemic increase was determined by the spread of a foreign bacterial clone or not. Among the 150 isolates relative to the aforesaid two periods, 12 different phage types (PTs) were observed. PT4 was the most common phage type among the strains isolated in 1980-1984 (61%) as well as in those of the epidemic period 1990-1993 (72%). PT8 was the second most frequent (33%) phage type in 1980-1984. It was substituted by PT1 (19%) in the 1990-1993 period. Analysis of rDNA patterns obtained after Hinc II digestions and Escherichia coli rRNA hybridizations showed 8 different patterns, A to H. The great majority of the strains studied (140 isolates, 93%) belonged to the ribotype A, showing a similar frequency both in 1980-1984 (36 of 39, 92%) and in 19901993 (104 of 111, 94%). The predominance of PT4 and ribotype A among both preepidemic and epidemic strains is in agreement with the hypothesis that host genetic diversity decline and modern farming practices in the poultry industry have facilitated a widespread dissemination of preexisting endemic strains. This hypothesis urges to plan new strategies in preventing S. Enteritidis infections.     

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A cluster of infections caused by Enterobacter cloacae was observed among preterm neonates in a neonatal intensive care unit (NICU) of a pediatric hospital in Osnabrück, Germany. The presence of similar antimicrobial susceptibility patterns among the bacterial isolates prompted an investigation to determine whether a limited spread of a single strain existed. All 12 E. cloacae isolates from the NICU and 50 nonrelated strains were fingerprinted by small-fragment restriction endonuclease analysis (SF-REA) of EcoRI DNA digests. Selected isolates were further characterized by pulsed-field gel electrophoresis (PFGE) of NotI- or XbaI-generated genomic restriction fragments. Epidemiologically unrelated strains were clearly discriminated by both methods. Results achieved by SF-REA and PFGE revealed that of the 12 isolates from the NICU, 11 belonged to the same genotypic cluster. Since all reagents and equipment for both techniques are commercially available, DNA fingerprinting by SF-REA or PFGE is proposed as a useful tool in the microbiology laboratory for investigating the epidemiological relatedness of E. cloacae strains of clinical and environmental origin.