Fifty verocytotoxin (VT)-producing Escherichia coli (VTEC) strains of serogroup O157 were characterized by phage typing, polymerase chain reaction (PCR) for VT genes and the E. coli attaching and effacing (eae) gene, and random amplified polymorphic DNA–PCR (RAPD–PCR) fingerprinting. The collection represented isolates obtained from patients with diarrhoea-associated haemolytic-uraemic syndrome (D+ HUS) and their family contacts, isolated in the Netherlands, Belgium and Germany between 1989 and 1993. Based on isolates from separate families (n = 27) seven different phage types were identified, types 2 (44%) and 4 (33%) were predominant. Eighty-five percent of the strains contained only VT2 gene sequences and 15% both VT1 and VT2. All strains of the dominant phage types 2 and 4 carried the VT2 gene. Strains that belonged to the minor phage types 8, 14, 32 carried both VT1 and VT2 genes, with the exception of two isolates identified as phage types 49 and 54 which contained only VT2 genes. All O157 VTEC strains possessed the chromosomally-located eae gene, which indicates its usefulness as virulence marker. RAPD–PCR fingerprinting identified four distinct banding patterns, with one profile found among 79% of the strains. Based on the combined results of all typing methods used in this study, the collection of 50 O157 VTEC strains could be divided into nine distinct groups. Strains isolated from different persons within one family could not be distinguished by any of these methods. The data suggest that O157 VTEC strains are members of one clone that has become widely distributed.

In this study we have evaluated the ability of three typing methods, pulsed field gel electrophoresis (PFGE), phage-typing and ribotyping, to discriminate not only between strains of differing serotypes but also between strains within a single serotype, heat stable serotype 2 (HS2). Forty-five isolates derived from cases of campylobacter enteritis occurring in the Cardiff area were examined. These included 18, mostly HS2, strains associated with an outbreak. The typing results for these and a further 39 epidemiologically unrelated strains of serotype HS2 were compared. This is the first report documenting the use of PFGE in an epidemiological investigation of Campylobacter jejuni in the UK. The results presented suggest that this technique is the most discriminatory of the three subtvping methods examined.

The possibility that a devastating human pandemic could arise, causing massive loss of human life, is discussed. Such a major threat to the human species is likely to be a virus, and would spread by the respiratory route. It need not necessarily cause massive loss of life, but if it caused serious illness or incapacity it would still have a major impact. A possible source is from an existing respiratory pathogen, but it would more probably arise from an infection that is maintained in an arthropod or vertebrate host, but which at present either does not infect humans, or if it does it fails to be effectively transmitted between them. More research should therefore focus on the pathogenetic factors and the viral determinants that promote respiratory transmission.

The aetiology of sporadic campylobacter infection was investigated by means of a multicentre case-control study. During the course of the study 598 cases and their controls were interviewed.

Conditional logistic regressional analysis of the data collected showed that occupational exposure to raw meat (odds ratio [OR] 9·37; 95 % confidence intervals [CI] 2·03, 43·3), having a household with a pet with diarrhoea (OR 2·39; CI 1·09. 5·25), and ingesting untreated water from lakes, rivers and streams (OR 4·16; CI 1·45. 11·9) were significant independent risk factors for becoming ill with campylobacter. Handling any whole chicken in the domestic kitchen that had been bought raw with giblets, or eating any dish cooked from chicken of this type in the home (OR 0·41–0·44; CI 0·24, 0·79) and occupational contact with livestock or their faeces (OR 0·44; CI 0·21, 0·92) were significantly associated with a decrease in the risk of becoming ill with campylobacter.

We describe the first cluster of cases of necrotizing fasciitis (NF) in this century in the United Kingdom (UK). Between 1 January and 30 June 1994 there were six cases (five confirmed, one probable) of Streptococcus pyogenes NF in west Gloucestershire, population 320000. Two cases died. The first two patients probably acquired their infections during the course of elective surgery performed in the same operating theatre, possibly from a nasopharyngeal carrier amongst the theatre staff. The remaining infections were community-acquired. Of 5 S. pyogenes isolates there were 2 M1 strains, 1 M3, 1 M5 and 1 M non-typeable strain. S. pyogenes NF had not been recorded in west Gloucestershire in the preceding 10 years and the incidence of S. pyogenes bacteraemia in England and Wales had not risen in the past 5 years.

The two presumably theatre-acquired infections raised several issues. The need for detailed bacteriological investigation of all cases of post-surgical NF was confirmed. Clusters of S. pyogenes infection following surgery should be managed by closure of the operating theatre until all staff have been screened for carriage. Closure of an operating theatre and screening of staff following a sporadic case is probably not justified because of the infrequency of surgical cross-infection with S. pyogenes. Regular, routine screening of theatre staff is neither practical nor necessary.

Following a buffet meal served to six guests at a private domestic function, five of the guests and the host developed symptoms of food poisoning. Salmonella enteritidis phage type 4 (PT4) was isolated from all four individuals who submitted faecal samples for investigation. Leftover samples of a savoury rice dish consumed by all six ill persons contained 6×103/gm Salmonella enteritidis PT4. The rice salad comprised boiled rice, raw carrots, eggs, cheese and curry powder. The curry powder and remainder of the pack of six eggs were negative on microbiological analysis. The rice dish had been prepared by heating in a 500 W microwave oven with a rotating turntable on full power for 5 min. Although the hazards of inadequate microwave cooking are well recognized, this is only the second outbreak of food poisoning from microwave cooking to be reported.

We have used molecular techniques to characterize 51 group A streptococci from Scotland and 17 ‘serious disease’ isolates from other countries, in order to establish the clonal structure of invasive Streptococcus pyogenes strains circulating between 1986 and 1993. Strains were grouped by restriction endonuclease analysis, pulsed field gel electrophoresis and ribotyping patterns, and were examined for the presence of alleles of the speA gene by polymerase chain reaction and DNA sequence analysis. Serious and fatal infections in Scotland were caused by several clones. One clone (9 of 51 strains) was M type 1 and possessed the speA gene allele 2. This was the clone previously identified as causing severe infection in the USA. Another clone (5 of 51 strains) was M type 3 and had speA gene allele 3. In view of the clear association of more than one clone with severe, invasive and fatal infections, horizontal gene exchange between genotypes merits further investigation.

During March and April of 1992, the health surveillance system began detecting increasing numbers of cases of meningococcal disease (MCD) in the Islamic holy city of Makkah (Mecca). We identified 102 bacteriologically confirmed cases (CC) and 80 suspected cases (SC) of MCD. Neisseria meningitidis was identified as Group A. III-1 clone. The ratio of male:female cases was 2·9:1. All age groups of males were affected. There was only one case among women aged 10–30; 50% of the adult female cases were 55 or older. The case-fatality ratio (CFR) was 14·7% among CC. Pakistanis, who comprised about one-third of the CC. had a CFR of 26·7%. Fifty-nine percent of CC were religious visitors. CC in residents were most common in persons living near the Holy Mosque (Haram), where the carriage rate reached 86%. A mass vaccination program against MCD was instituted, using AC bivalent meningococcal vaccine (MCV). An abrupt drop, from a mean of 15 CC per week to 2 CC per week (only in visitors), coincided with vaccinating 600000 persons over 2 weeks. Makkah residents who had been vaccinated against MCD were less likely to have contracted MCD (OR = 0·17, 95% CI: 0·06–0·50). MCV was of no significant protective value if it had been administered 5 years before the outbreak. The main reason for not being vaccinated as stated by both cases (71%) and controls (45%) was not knowing about the disease. The age and sex differences probably relate to differences in exposures to crowded conditions. Health education should illuminate the seriousness of the disease and the importance of vaccination.

Data from volunteers challenged with Salmonella typhi were reanalysed to explore the relationship between challenge dose and severity of disease. Among 120 ill volunteers who received between 105 and 109 organisms, dose was weakly correlated with peak temperature (r = 0·22, 95% CI 0·04–0·39), duration of temperature above 103 °F (39·4 °C: r = 0·13, 95% CI –0·03 to 0·55) and symptom score (r = 0·27, 95% CI 0·09–0·43). The association with symptom score was lost after adjusting for year, and the findings depended on the definition of illness used: with stricter definitions the associations with temperature were also lost. The study shows the need for caution in interpreting the relationship between dose and severity of disease.

Vaccination against Neisseria meningitidis is not part of routine immunization schemes in any country; instead, targeted vaccination of groups at the highest risk is recommended during outbreaks and epidemics. After a long period of sporadic occurrence of meningococcal invasive disease, a new clone of Neisseria meningitidis C: 2a: P1.2, ET-15/37, occurred in the Czech Republic, and caused local outbreaks in two neighbouring districts, Olomouc and Bruntal, in spring 1993. In Olomouc, a mass campaign was conducted during which 6191 students were vaccinated (5·6 % of the total population of this locality and 96% of all students in the age group 15–19) within 2 weeks in June 1993. In Bruntal district, no such campaign was organized. In Olomouc, the incidence of invasive disease caused by Neisseria meningitidis C in the age group 10–24 decreased from 57 to 0 per 100000 (P < 0·001) during the post-vaccination period (July 1993–August 1994), but no such decrease was observed in Bruntal. Although other factors can affect the frequency of disease, these results support the current recommendations of targeted vaccination in outbreaks of meningococcal disease.

The genetic and biochemical basis of ampicillin resistance amongst the aerobic Gram-negative commensal faecal flora of healthy volunteers in South Africa has been determined. Amongst 608 ampicillin resistant strains isolated from 320 of the participants, 158 were able to transfer their ampicillin resistant determinants into Escheriehia coli K-12 J62–2. Iso-electric focusing of the β-lactamases, extracted from the transconjugants, demonstrated that ampicillin resistance resulted from the presence of the TEM-1, TEM-2 and SHV-1 β-lactamases in 94·3%, 2·5% and 3·2% of isolates respectively. Endonuclease restriction digests of the plasmids isolated from the transconjugants showed that the β-lactamase genes were present on a wide variety of plasmid types; 101 distinct plasmid endonuclease restriction patterns were identified. Transferable ampicillin resistance was associated with resistance to other antibiotics at the following frequencies: trimethoprim (48·7 %), streptomycin (35·4 %), tetracycline (27·2%), spectinomycin (9·5%), chloramphenicol (3·2%) and gentamicin (1·3%). One antibiotic resistance pattern, ampicillin and trimethoprim. predominated (28%). In total. 77·9% of the plasmids conferred resistance to other antibiotics raising the possibility that use of any of these agents, not simply ampicillin, may contribute to the maintenance of resistance genes.

During July 1991, a single laboratory reported an increased number of an unusual salmonella isolate. An outbreak control team was convened. A case was defined as an individual with diarrhoea from whose faecal sample Salmonella hadar was isolated after 1 July 1991. By 30 July, 90 isolates had been identified and 57 persons interviewed including 39 primary cases. Interviews failed to identify any common features among the cases. A review of the laboratory procedures revealed that the selenite enrichment medium was inoculated using the spoon from the stool collection kit after it was used to emulsify the faecal sample with saline for microscopy. Salmonella hadar was isolated from this saline. Once this practice was stopped, no further isolates of S. hadar were made. This pseudo-outbreak is a powerful reminder to verify the existence of an outbreak, especially when epidemiological data are inconsistent.

A total of 332 Aeromonas spp. originating from drinking water (n = 75), fresh water (n = 57), chicken and ground beef (107), human faecal samples in association with travelling (n = 49), human faecal samples not associated with travelling (n = 38), and six strains from human blood cultures were studied by phenotypic methods and by using analysis of ribopatterns as a molecular method for the identification of the 13 known hybridization groups (HGs). Also included were the reference strains of each HG. A. hydrophila HG 1, A. caviae HG 4 and A. veronii biotype sobria HG 8/10 were the most important genospecies identified in human faecal samples. A. hydrophila HG 2 and A. media HG 5B predominated in drinking water and A. hydrophila HG 2 and HGS, A. media HG 5A and HG 5B predominated in fresh water. In drinking water only one isolate was A. hydrophila HG 1 and two isolates were A. caviae HG 4. Clinically important Aeromonas spp. HG 1 (A. hydrophila), HG 4 (A. caviae) and HG 8/10 (A. veronii biotype sobria) were common in chicken and ground beef. In contrast to the drinking water samples, HG 5A was common in chicken and ground beef samples. Atypical, unidentified isolates were most often found in fresh water samples (12/57 strains). Although water has been suspected of being an important source of human aeromonas infections, clinically important HGs were found to be in the minority among Aeromonas spp. identified in drinking water or fresh water. The distribution of Aeromonas spp. HGs among drinking water, chicken and ground beef samples was also different, suggesting that contamination of meat or chicken may not originate from water.

A molecular typing method for Staphylococcus aureus based on coagulase gene polymorphisms (coagulase gene typing) was evaluated by examining a total of 240 isolates which comprised 210 methicillin-resistant S. aureus (MRSA) and 30 methicillin-susceptible S. aureus (MSSA) collected from a single hospital. By Alul restriction enzyme digestion of the PCR-amplified 3′-end region of the coagulase gene including 81-bp repeated units, the MRSA and MSSA isolates examined were divided into 6 and 12 restriction fragment length polymorphism (RFLP) patterns, respectively, whereas five patterns were commonly detected in MRSA and MSSA. MRSA isolates that showed a particular RFLP pattern were considered to be predominant in the hospital. Coagulase typing with type-specific antisera was also performed for all S. aureus isolates for comparison. Coagulase types II and VII were most frequently detected and included isolates with four and five different AluI RFLP patterns, respectively, whereas each of the other coagulase types corresponded to a single RFLP pattern. These results indicated that RFLP typing was more discriminatory than serological typing, for typing S. aureus and demonstrated its utility in epidemiologic investigation of S. aureus infection in hospitals.

In a recent survey of trimethoprim resistance. 357 Gram-negative aerobic organisms were isolated from healthy volunteers from rural and urban populations in South Africa. Trimethoprim resistance did not transfer to an Escherichia coli J62–2 recipient strain by conjugation in a liquid mating in 161 (45·1%) of the isolates. These isolates which did not transfer their resistance were probed with intragenic oligonucleotide probes for the types Ia. Ib. IIIa. V. VI. VII. VIII. IX. X and XII dihydrofolate reductase genes. Contrary to all previous data, the most prevalent dihydrofolate reductase gene in this group of non-transferable isolates which hybridized, was the type VII (38%) followed by the type Ia (25%). Ib (12%). V (1·7%) and VIII (1·2%). None of the strains hybridized to the types IIIa. VI. XI. X and the XII dihydrofolate reductase probes. Southern blots of plasmid and chromosomal DNA from selective isolates revealed that the type VII dihydrofolate reductase genes were located on the chromosome and were associated with the integrase gene of Tn21. However, the type Ib and V dihydrofolate reductase genes were all found on plasmids which could not be mobilized. The type Ia dihydrofolate reductase genes were found on both non-transferable plasmids and on the chromosome. The nature of the genetic structures associated with a dihydrofolate reductase gene strongly affects the means of spread of the gene in a population.

Sixty-nine strains of Vibrio cholerae 01 isolated at different times were analysed to investigate if there were any differences among the 01 strains isolated before, during and after the advent of the O139 serogroup. Of the 69 01 strains examined, 68 belonged to the Ogawa serotype while one belonged to the Inaba serotype. With the exception of one strain all other strains of V. cholerae 01 belonged to the eltor biotype. A single O1 strain isolated before the emergence of the O139 serogroup could not be classified as either eltor or classical biotype because it was resistant to both classical and eltor specific bacteriophages. Marked variations in the susceptibility to antibiotics of V. cholerae O1 isolated during the different periods were observed. In addition, strains of V. cholerae isolated after the epidemic of serogroup O139 in Calcutta showed an expanding R-type with resistance to a variety of drugs as compared to the 01 strains isolated before the advent of the O139 serogroup. From this study, it is clear that there is a substantial mobility in genetic elements of V. cholerae Ol which necessitates a continuous monitoring to keep abreast of the changing traits of the etiologic agent of cholera.

PCR-based DNA fingerprinting was used to characterize 48 clinical isolates of Yersinia enterocolitica. The samples were examined by random amplified polymorphic DNA (RAPD-PCR) and inter-repeat PCR (IR-PCR). IR-PCR with two enterobacterial repetitive intergenic consensus primers resulted in patterns which were poorly discriminated; 2 of 11 arbitrary primers (RAPD-PCR) provided sufficient discriminatory power. In comparisons with serotyping and biotyping, RAPD-fingerprinting was the most discriminatory technique and may therefore be a valuable epidemiological tool for the study of Y. enterocolitica infections.

The present study was undertaken to investigate the frequency of the nasal carrier rate of Staphylococcus aureus. The investigation was performed on 104 healthy persons. The total number of swabs performed was 1498 and this resulted in isolation of 522 S. aureus strains. All strains have been identified, tested for antibiotic susceptibility, and phage-typed. The carrier-index (number of positive swabs/number of total swabs for each individual person) was compared with different sampling and culturing methods, phage type, age, and resistance to antibiotics. There was statistical difference in carrier rate according to sex (P ·05). Among the 104 persons 15 (14·4%) were persistent carriers, 17 (16·3%) intermittent carriers, 55 (52·9%) occasional carriers and 17 (16·3%) non-carriers. Among intermittent and occasional carriers the phage-type distribution was different from the S. aureus strains isolated from Danish hospitalized patients in 1992, while the persistent carriers had similar phage-type distribution.

An outbreak of infections due to multiple antibiotic-resistant bacteria took place over a period of approximately 18 months in a renal unit. Strains of Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, Citrobacter spp. and Pseudomonas spp. were involved, and a variety of antibiotic resistances was encountered. Closely related plasmids encoding resistance to aztreonam, ceftazidime and piperacillin, possibly derived from an archetypal plasmid of 105 kb were found in the majority of isolates examined. After limiting the use of aztreonam the incidence of new patient isolates of multiple-resistant organisms was greatly reduced. This study demonstrated how molecular studies can contribute to the control of an outbreak situation in a hospital unit by providing an impetus to reduce the use of specific antibiotics.

An outbreak of cryptosporidiosis which affected 44 people in January and February 1991 was identified through local surveillance at a South London Public Health Laboratory. Preliminary enquiries revealed that more than half the patients were adult and that there were no common factors other than geographical association. A case-control study showed a significant association between illness and consumption of tap water supplied by a particular water company, as well as a dose response effect. There were no apparent breaches or irregularities in the water distribution system and no indication of a problem through routine monitoring indices. This incident demonstrates the problems of establishing the source of cryptosporidiosis outbreaks in the absence of evidence of environmental abnormality, as well as possibly indicating that water conforming to current treatment standards may occasionally contain sufficient numbers of Cryptosporidium oocysts to cause sporadic cases or clusters.