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.

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.

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.

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.

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.

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.

During October 1992 an increase in the number of isolates of Salmonella mikawasima, a rare serotype, was noted including a cluster of nine cases in the South West Thames region. A case control study was conducted and univariate analysis showed a statistical association between illness and eating at takeaway A for cases compared with household controls (P = O003) and with neighbourhood controls (P = 0.0245). Cases were also more likely to have eaten kebabs than were controls or average takeaway A customers, implicating doner kebabs as the most likely vehicle of infection. Plasmid profile analysis of the nine cases' isolates showed them to be indistinguishable and to be characterized by a single plasmid of approximately 60 MDa.

The original source of the Salmonella mikawasima contamination was not determined, but food preparation practices for kebabs at takeaway A were insufficient to protect against illness if contaminated. This outbreak was only recognized because of the unusual serotype, but could be an indication of a more widespread problem with doner kebabs.

The fact that Helicobacter pylori can revert to a coccoid form has stimulated speculation about its role in transmission and as a possible cause of reinfection in duodenal ulcer disease. Bismuth subcitrate (32 μg/ml), bismuth subsalicylate (64 μg/ml), amoxicillin (0·05 μg/ml) and erythromycin (4 μg/ml) inhibited the growth of H. pylori and stimulated the formation the formation of basically respiring but non-culturable coccoid structures. The presence of polyphosphates as energy and phosphorus source permits a certain level of endogenous metabolism to preserve RNA and DNA, as well as structural components like cell wall, cell membrane and cytoplasma for at least 3 months. However, the applied standard laboratory methods were insufficient for regrowth of H. pylori out of the coccoid form.

Sixty-five strains of Acinetobacter baumannii which had been isolated from patients and the indoor environment of a neonatal intensive care unit and, for comparative purposes, isolates from three other wards, were examined by means of electrotyping and analysis of whole-cell protein and antibiotic resistance patterns. Fourteen different electrotypes were determined. The predominant type, a multiply resistant acinetobacter clone, persisted in the neonatal ward over several months. The results underline the usefulness of electrophoretic subtyping, in particular by means of allozyme pattern and as a supplement to whole-cell protein pattern analysis, in epidemiological investigations into the routes of transmission of nosocomial A. baumannii infections.

Epidemiological evidence suggests that monoclonal antibody type 2 positive (MAB 2+) Legionella pneumophila serogroup 1 (LP1) more often causes disease than do MAB 2− isolates, and there is evidence that MAB 2− LP1 grow less well in cells than do MAB 2+ bacteria. We tested the intracellular growth rates of ten randomly selected MAB 2− LP1 isolates, by using guinea-pig alveolar macrophages, and human monocyte-derived macrophages. Save a low virulence control, all ten MAB 2− isolates grew as well in cells as a virulent MAB 2+ isolate. Heterogeneity of MAB 2− LP1 growth in cells exists, making poor intracellular growth an unlikely explanation for why MAB 2+ LP1 appear to cause disease more often.

A mathematical model was used to estimate malaria transmission rates based on serological data. The model is minimally stochastic and assumes an age-dependent force of infection for malaria. The transmission rates estimated were applied to a simple compartmental model in order to mimic the malaria transmission.

The model has shown a good retrieving capacity for serological and parasite prevalence data.

An alternative way to estimate the endemic level of malaria amongst Brazilian indians is proposed. This is achieved by estimating the age-related “force of infection’ of malaria (the effective inoculation rate), applying a mathematical model, described elsewhere, to serological data. In addition we present a way to estimate the Basic Reproductive Rate of malaria in the same area. The results have shown a good degree of accuracy in describing the endemic pattern of malaria in the area, and also indicate some relevant aspects of its age distribution related to the design of control strategies.

Four influenza type B viruses isolated in Russia during periods of relatively low (1987–8) or high (1990–1) influenza B activity were characterized antigenically using a microneutralization assay. These isolates were antigenically similar to contemporary reference strains from either of two separate lineages represented by B/Victoria/2/87 and B/Yamagata/16/88. The evolutionary relationships of the variable portion of the haemagglutinin (HA1) genes of these viruses were determined by comparison with influenza B HA1 sequences previously obtained. The Isolate B/USSR/2/87, collected during the 1987–8 influenza season, was found to be closely related to viruses on the B/Victoria/2/87 lineage that circulated during the 1988–9 influenza season in the United States. Sequence analysis of the isolates from the 1990–1 influenza season demonstrated co-circulation of viruses from both the B/Victoria/2/87 and B/Yamagata/16/88 lineages in Russia, confirming the antigenic analysis.

IgM antibody response to Crimean-Congo haemorrhagic fever (CCHF) virus was monitored in experimentally infected sheep and cattle by an IgM capture enzyme-linked immunoassay (ELISA). Specific binding of antigen was detected by a rabbit anti-CCHF horseradish peroxidase conjugate or a sandwich technique with hyperimmune mouse anti-CCHF ascitic fluid and commercially available anti-mouse immunoglobulin peroxidase conjugate. The persistence of IgM antibody activity was found to be of shorter duration than in humans, and this may be a function of the relative lack of susceptibility of these animals to infection with CCHF virus. IgG antibody responses in the sheep could be monitoried by sandwich ELISA using commercially available anti-sheep immunoglobulin peroxidase conjugates. Total antibody activity in the sera of experimentally infected sheep, cattle and small mammals could be monitored in a competitive ELISA (CELISA) using rabbit anti-CCHF peroxidase conjugate. The CELISA was applied to the sera of 960 wild vertebrates from a nature reserve in South Africa, and the prevalence of antibody was found to be greatest in large mammals such as rhinoceros, giraffe and buffalo, which are known to be the preferred hosts of the adult tick (Hyalomma) vectors of the virus.

Between 1989 and 1992, 7970 wild ungulates, comprising 14 different species, were tested for antibodies to types SAT 1, SAT 2 and SAT 3 foot-and-mouth disease (FMD) virus. Of these 1·2% were found to be positive and these included impala (Aepyceros melampus), eland (Taurotragus oryx), waterbuck (Kobus ellipsiprymnus) and sable (Hippotragus niger). All the positive animals were either from the wildlife areas where buffalo (Syncerus caffer) occur or from ranches where clinical FMD had occurred in cattle. The role of these animal species in the current epidemiology of FMD in Zimbabwe is discussed.

The epidemic cycle of Chlamydia pneumoniae infection in eastern Finland, 1972–1987 Fig 2. For the top right-hand graph of the four shown, the legend should show: urban. … rural, –––; all, ——. The legend for the remaining three graphs is correct.