Sixteen cases of verotoxin producing Escherichia coli (VTEC) O 157:H7 Phage Type 49 infection were identified in the North West of England from 1 September to 1 November 1991, eight of whom lived in or around the same large town. Eleven of the cases were aged 10 years or less, and five of the affected children developed haemolytic uraemic syndrome. A case control study demonstrated a strong association between VTEC O 157: H7 PT 49 infection and the consumption of a locally produced live yoghurt. This is the first time that an outbreak of VTEC O 157 infection has been linked to the consumption of yoghurt and this vehicle of infection should be considered when investigating such outbreaks in future.

Ribosomal RNA gene patterns, randomly amplified polymorphic genomic DNA (RAPD) profiles and plasmid profiles were used to discriminate between 28 strains of Campylobacter jejuni serogroups O1 and O2 (Penner). Most isolates were biotype I (Lior). The strains were representative isolates from a UK school outbreak of enteritis (7 cases) and from 21 sporadic human cases of enteritis in 4 countries. The molecular techniques discriminated to various degrees between strains in each of the serogroups. The outbreak strains were homogeneous in most molecular features but a variety of types was detected amongst the isolates from the sporadic cases. Five groups of two or more strains with identical ribopatterns were identified and within each, strains from different patients were homogenous with respect to serogroup. RAPD profile typing based on numerical analysis generally matched ribotyping. Plasmid profiling overall gave least discrimination but was useful in separating some strains similar in other features. We concluded that optimal discrimination of C. jejuni could best be achieved using a combination of phenotypic and genotypic properties. Hae III ribotyping was the single most discriminatory and reproducible technique investigated. Several strains of C. jejuni from sporadic infections had similar molecular profiles which have potential for general typing purposes.

This study investigates the course of tuberculosis in a naturally infected badger population, its impact on the population and the risk of spread to other species in the light of capture data and post-mortem findings from 47 tuberculous badgers, stratified by age group and sex, accrued since 1975. The findings are compared with those for 260 badgers from the same population in whom no evidence of infection was detected. Detailed estimates of seasonal variations in bodyweight for uninfected male and female cub, yearling and adult badgers are presented and compared to the weights at post-mortem examination of the tuberculous badgers, in whom poor condition and weight loss were the principal presenting signs. Lesions were seen especially in the lungs and associated lymph nodes, and in the kidneys. Organisms were detected intermittently in faeces, urine, sputum and discharging bite wounds. Infected animals could survive for nearly 2 years and produce cubs successfully.

Some strains of Escherichia coli belonging to serogroups O26, O55, O111 or O128 produce Vero cytotoxin (VT). These serogroups are included in the range of enteropathogenic E. coli (EPEC) serogroups for which commercial antisera are available. In an attempt to obtain information on VT-producing strains other than those of serogroup O157, 122 strains belonging to these four serogroups and isolated in 1991 from patients with diarrhoea in the United Kingdom were tested for hybridization with VT probes. Only 18 of the 122 strains were VT-positive and these were O26 or O128. However 90 strains hybridized with the E. coli attaching and effacing (eae) probe (including 14 VT-positive strains) and 17 with the enteroaggregative E. coli (EAggEC) probe. For 78 eae-positive and 9 EAggEC-positive strains, tissue culture tests correlated with the probe results as the strains gave, respectively, either localized adhesion and a positive fluorescent-actin staining test or a characteristic aggregative attachment. A total of 111 of the 122 strains belonging to serogroups O26, O55, O111 or O128 possessed properties that may be associated with the ability to cause human diarrhoeal disease, and similar studies are needed on strains from the other classical EPEC serogroups.

For Salmonella typhimurium from humans in England and Wales, the incidence of multiple resistance more than doubled over the 8-year period 1981–8 and, over the next 2 years, increased by a further 7%. From 1981 to 1988 both resistance and multiple resistance also increased significantly in S. virchow and although multiple resistance did not increase over the next 2 years, the overall incidence of resistance has continued to rise. In 1990 the majority of S. typhimurium from cattle were multiply-resistant and the occurrence of such resistance has quadrupled since 1981. Multiple resistance has also increased in S. typhimurium from pigs and, to a lesser extent, from poultry. In contrast, multiple resistance has remained uncommon in the poultry-associated serotype S. enteritidis. For S. virchow, multiple resistance was common in a phage type frequently associated with poultry meat imported from France.

The continuing use of a range of different antimicrobials in calf husbandry has been an important factor in promoting the emergence of multiply-resistant strains of S. typhimurium in cattle. In contrast, multiple resistance has remained rare in those serotypes associated with poultry, where the use of such antimicrobials has been less intensive.

It is hoped that recent recommendations discouraging, in veterinary medicine, the prophylactic use of antibiotics with cross resistances to those used in human medicine will result in a reduction in the occurrence of multiresistant strains in food animals and subsequently in humans.

In May-June 1992 cases of infection with verocytotoxin-producing (VT+) Escherichia coli O157 in South Yorkshire could have been associated with prior consumption of beef from a local abattoir. During investigation of the abattoir, bovine rectal swabs and samples of meat and surface swabs from beef carcasses were examined for E. coli O157, isolates of which were tested for toxigenicity, plasmid content and phage type. E. coli O157 was isolated from 84 (4%) of 2103 bovine rectal swabs; of these 84, 78 (93%) were VT+, the most common phage types being 2 and 8, the types implicated in the cluster of human cases. Positive cattle were from diverse sources within England. E. coli O157 was isolated from 7 (30%) of 23 carcasses of rectal swab-positive cattle and from 2 (8%) of 25 carcasses of rectal swab-negative cattle. The study has shown that cattle may be a reservoir of VT+ E. coli O157, and that contamination of carcasses during slaughter and processing may be how beef and beef products become contaminated and thereby transmit the organism to man.

The Phene Plate system for typing Salmonella serotypes (PhP-S) is a simple automated typing method based on biochemical fingerprinting. It gives a quantitative value of the metabolism of various substrates by measuring the speed and intensity of each reaction. The ‘biochemical fingerprint’ of each isolate is used to calculate similarities among the tested strains with a personal computer program. We used this system to examine a collection of 86 strains of Salmonella enteritidis isolated from human sporadic cases in Germany between 1980 and 1992. Twenty-three biochemical phenotypes (BPTs) consisting of 9 common (C) and 14 single (S) BPTs were identified. BPTs C2 and C4 containing 20 and 36 strains respectively accounted for 65% of the isolates. Strains of BPT C2 were found over a wide period of time whereas strains of BPT C4 were isolated during the period between 1988 and 1992. With phage typing, 11 discrete phage types (PTs) and 18 strains designated as non-specific type (NST) were identified. PTs 4 and 8 with 39 and 17 strains respectively were the dominant PTs. Strains of PT 8 were isolated over a wide period of time whereas all (except one) strains of PT 4 were isolated between 1988 and 1992. Combination of biochemical fingerprinting and phage typing divided the strains into 25 phenotypes (BPT:PTs). Whilst phenotype C2:8 was found over a number of different years, phenotype C4:4 was isolated only between 1988 and 1992. These findings indicate the presence of one persistent and one recently emerged phenotype among S. enteritidis strains in Germany. Although both methods identified the presence of both major and less common types, biochemical fingerprinting with the PhP-S system also provided information about relationships between the strains.

This paper describes a matched case-control study to determine the efficacy of BCG vaccine in preventing the occurrence of leprosy in southern Malaŵi, a previously unstudied area. The BCG immunization rate amongst 145 individuals with leprosy was 44.8%, compared to 62.5% in 290 matched controls. The protective efficacy of BCG vaccine against leprosy in this region was estimated to be 63.6%; smallpox immunization had no effect. These findings support the view that BCG vaccine should be considered as a control measure in areas where leprosy is endemic.

Strains of Salmonella enteritidis belonging to phage type 4 (SE4) were grown in the peritoneal cavities of chickens, and without subculture on laboratory media examined for inducible in vivo phenotypic characteristics. These bacteria expressed three major outer membrane proteins (OMPs) of 33, 35 and 36 kilodaltons (kDa), and iron regulated OMPs of 74, 78 and 81 kDa. Bacteria growing in vivo did not express flagella, or fimbriae with a subunit molecular mass of 14 kDa (14 kDa fimbriae). Two OMPs of 55 and 23 kDa, expressed during culture in nutrient broth, were repressed during growth in chickens. Possession of a 38 MDa ‘mouse virulence’ plasmid did not influence the expression of OMPs, flagella or fimbriae. It was concluded that strains of SE4 growing in chicken tissues, use an enterobactin mediated iron uptake system to obtain ferric ions, do not express flagella or 14 kDa fimbriae and appear not to express novel OMPs involved in survival in vivo.

The growth of Salmonella enteritidis PT4 in albumen around an intact yolk was governed by the age of the egg on inoculation. In the majority of eggs, held at 20° C, the bacterium was unable to grow rapidly until eggs had been stored for approximately 3 weeks. The multiplication of S. enteritidis in stored eggs appeared to be associated with alterations to the yolk membrane which allowed the bacterium to either invade the yolk or obtain nutrients from it. The rate at which egg contents change to permit the growth of S. enteritidis would appear to be temperature related and took place more rapidly when eggs were stored under conditions where temperatures fluctuated and, on occasions, reached 30° C.

Rates of acquisition and mean duration of nasal carriage of different serotypes of Streptococcus pneumoniae have been estimated by fitting a stochastic model to longitudinal carriage data in children from Papua New Guinea. Immunogenicity and two indices of relative invasiveness were determined for each serotype. Immunogenic serotypes were less frequently acquired and were carried for shorter periods, but no relationship between immunogenicity and invasiveness was apparent using either index of invasiveness. Frequent invasion was associated with a high acquisition rate and high frequency and prolonged duration of carriage. Carriage studies can provide a broad indication of which serotypes cause invasive disease but not the proportion of disease due to individual serotypes; some serotypes which cause invasive disease (e.g. serotype 46) are not found even in extensive carriage studies. The antibiotic resistance of carriage organisms, however, does approximate the resistance patterns of invasive organisms and thus may be used to monitor changing patterns of antimicrobial susceptibility in the community.

A quick, semi-quantitative method of detecting Salmonella species which contain the virulence plasmid has been developed using the polymerase chain reaction (PCR). A pair of primers have been synthesized encompassing a 500 bp fragment of the spvR virulence gene. Competitor DNA consisting of the spvR gene with a 94 bp deletion situated between the primer recognition sequences, was cloned into a plasmid vector. Co-amplification of the ‘unknown’ target salmonella DNA with known quantities of competitor DNA in the same reaction tube gave PCR products of 500 and 406 bp respectively. Visual assessment of the ratio of the two products on ethidium bromide stained agarose gels provided an estimate of the approximate number of salmonella cells present in avian faeces.

The technique could be applied to detect quantifiably any non-host DNA in clinical samples if a suitable DNA sequence for primer construction is available.

A national survey was conducted of 3150 notified cases of whooping cough in order to determine age-specific pertussis vaccine efficacy by the ‘screening’ method. The cases were collected over two periods, one just prior to the start and one at the first peak of the whooping cough epidemic of 1989–90. Vaccination status was determined by a postal questionnaire to the reporting doctor and clinical data were also collected to provide efficacy estimates according to standardized case definitions. Overall, observed vaccine efficacy was high but differed between epidemic (87%) and non-epidemic (93%) periods (P = 0·03). Efficacy estimates were generally higher for typical or severe cases than for children with an atypical illness. Vaccine efficacy declined with age (P < 0·01) but estimates remained high up to the age of 8 years. This study will provide baseline data for comparison with efficacy observed from similar studies of children immunized at an accelerated schedule and from phase III studies of acellular pertussis vaccines performed elsewhere.

Between January and March 1992, 361 faecal specimens were collected from the healthy black population in the Transvaal Province of South Africa. Each specimen was examined for the prevalence of antimicrobial resistance in commensal bacteria. Volunteers, from both rural and urban dwellings, were divided into four age groups. The overall carriage rate of resistance varied from 88.6% for ampicillin, 74.2% for trimethoprim, 52.6% for chloramphenicol, 10.2% for nalidixic acid to 7.5% for gentamicin. The carriage of resistance found to each individual antimicrobial agent was slightly higher in the rural population rather than the urban population but there was no correlation between the prevalence of antimicrobial resistance and the age group.

Escherichia coli and salmonella strains with plasmids conferring resistance to gentamicin and apramycin have been isolated with increasing frequency both from cattle and hospital patients in Belgium. The apramycin-gentamicin resistance plasmids were characterized in recipient strains by their profiles and molecular weights using agarose gel electrophoresis, by their antimicrobial resistance patterns and by replicon typing using a series of DNA probes specific for the genes controlling their systems of replication. Overall, most of the plasmids differed in their DNA electrophoretic patterns. Seventeen different antimicrobial resistance profiles were observed, and there were six different types of replicons. However, two replication genes predominated and had a preferential distribution in different bacterial species. The rep FIC.a plus rep Q multireplicon was found mainly in plasmids recovered from gentamicin- and apramycin-resistant E. coli while replicon of the type rep FIC.b largely prevailed in S. typhimurium. Identical replication genes were found in most animal and human strains, hence suggesting a high homology between apramycin- gentamicin plasmids in these communities. Finally, our results indicate that the rapid spread of apramycin-gentamicin-resistance in several species of Enterobacteriaceae isolated from animals and from humans in Belgium is not due to a single plasmid, but rather that the gene encoding AAC(3)-IV is carried by various replicons.

Geographically widespread outbreaks involving commonly isolated organisms and where the vehicles of infection are commonly eaten foodstuffs pose particular difficulties at a technical and organizational level.

An outbreak of Salmonella typhimurium infection, affecting 39 people, spread over a wide area in North West England and North Wales in April and May 1991, was detected thanks to the practice of sending specimens to the national reference laboratory where phage typing and characterizing of antibiotic resistance patterns enabled the identification of a cluster of distinctive isolates (S. typhimurium DT193 resistant to sulphonamides, trimethoprim and furazolidone). An investigation, involving twenty environmental health departments in addition to health authorities and the Public Health Laboratory Service, showed an association between the illness and eating loose sliced cooked ham (P = 0.004). Detailed tracing of the chain of supply of the ham showed this to be particularly cooked ham originating from a single small local producer (P = 0.00003). Further investigation of that producer revealed that a batch of ham distributed on one day in early April was undercooked due to a malfunction in cooking equipment.

A total of 17 isolates of Vibrio mimicus from patients, 29 from environment and 2 from food was examined for toxigenicity. Sixteen (94%) clinical isolates and one (50%) from food produced TDH-like toxin, whereas none of the environmental isolates did so. The food from which V. mimicus with TDH-like toxin production was isolated, was one which had caused food poisoning. Only one environmental strain produced CT-like toxin, whilst ST-like toxin was not detected from any strains tested.

Plasmid analysis of Salmonella enteritidis isolates from human gastroenteritis cases and from two commercial egg-producing poultry flocks was performed to determine if the poultry flocks were the source of the human infections. The plasmid profile and restriction fragment pattern (fingerprint) of five S. enteritidis isolates from human cases matched those of nine isolates from internal organs of egg-laying hens in one flock which was the source of eggs consumed by the cases. Another commercial flock was epidemiologically associated as the source of eggs consumed by affected persons in four separate gastroenteritis outbreaks from which S. enteritidis isolates were available. Five S. enteritidis isolates from human cases in these four outbreaks had the same profile and fingerprint, and they all matched those of the 24 isolates from hens in this flock. These results provide further documentation of egg-borne transmission of S. enteritidis to humans.

Methicillin-resistant Staphylococcus aureus isolates from an outbreak of 17 cases of wound infection in a municipal hospital were typed by conventional methods, phage typing by three sets of phages, reverse phage typing and plasmid profiles, as well as by genomic DNA fragment patterns obtained after Sma-I digestion and pulsed-field electrophoresis. These isolates were non-typable by phages, only some were typable by reverse phage typing and were not uniform in plasmid profile. Only the genomic DNA fragment patterns resulted in a clear discrimination of 2 strains (12 isolates for the first and 7 isolates for the second). Both strains were disseminated in different wards of the same hospital and one strain had obviously spread to another clinic in the same city.