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.

A correlation between childhood crowding and the later development of gastric cancer has been demonstrated by Barker and colleagues, who proposed that the relationship was the consequence of infection by an organism such as Helicobacter pylori. In order to test this hypothesis the presence of IgG antibodies to H. pylori in sera from blood donors in North Wales has been investigated. During donation sessions, donors answered questions relating to social conditions and domicile in childhood (at age 10 years) and adult life (the preceding 2 years).

A stepwise logistic regression analysis of the data demonstrated significant independent relationships between seropositivity and the following factors: sharing a bed in childhood, housing density, locality of birth, adult social class and age.

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.

An epidemiological investigation was conducted to identify risk factors related to hygiene and husbandry practices which determine the introduction of Campylobacter spp. into broiler chicken flocks. All 176 broiler farms in an area in southeastern Norway participated in the study. Each farm was represented by one flock selected at random during a one-year period. The flocks were examined for Campylobacter colonization at slaughter, and the flock managers were sub-sequently interviewed about hygiene and husbandry practices. Campylobacter spp. were recovered from 32 (18%) of the flocks. The proportion of colonized flocks varied geographically and seasonally with a peak in the autumn. The following variables were found to be independently associated with an increased risk of Campylobacter colonization using logistic regression analysis: (i) feeding the broilers undisinfected water (odds ratio (OR) = 3·42, P = 0·045), (ii) tending other poultry prior to entering the broiler house (OR = 6·43, P = 0·007), (iii) tending pigs before entering the house (OR = 4·86, P = 0·037), (iv) geographic region (Hedmark versus Østfold county) (OR = 2·91), P = 0·023, (v) season (autumn versus other seasons) (OR = 3·43, P = 0·008). Presence of rats on the farm was associated with an increased risk, but this factor did not reach statistical significance (OR = 3·96, P = 0·083). Preventive measures should include dis-infection of drinking water and strict hygienic routines when the farm workers enter the rearing room. The results indicated that disinfection of drinking water is the preventive measure most likely to have the greatest impact on the prevalence of Campylobacter among broiler chicken flocks in the study area (population attributable fraction = 0·53).

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.

The discriminatory power of four methods for typing of Listeria monocytogenes was compared. The four methods were multilocus enzyme electrophoresis (MEE), ribotyping, restriction enzyme analysis (REA), and a newly developed Danish phage typing system. Ninety-nine human clinical, food and slaughterhouse isolates of Listeria monocytogenes were typed by each method. The most discriminatory single typing method was phage typing with an overall discriminatory index (DI) of 0·88 followed by REA, MEE and ribotyping with DI-values at 0·87, 0·83 and 0·79 respectively. Considering strains from each of the two predominant O-serotypes alone, serotype 1 was best discriminated by the molecular typing methods, in particular REA, which showed a DI of 0·92. The serotype 4 strains were best discriminated by phage typing (DI = 0·78). If two or more typing methods were combined, the combination of REA and MEE were found to be the most discriminatory combination. The DI values were 0·96, 0·74 and 0·90 for serotype 1, 4, and both combined, respectively. Phage typing is a rapid and inexpensive typing method but not as reproducible as the molecular typing methods. It is the most suitable method for mass screening. In situations where results are required to be highly reliable, i.e. when studying the relationships between only a few strains, a single or a combination of molecular typing methods should be used, preferable MEE and REA.

It is generally accepted that most patients with Clostridium difficile-associated diarrhoea acquire the organism from the environment. Recently we demonstrated that household pets may constitute a significant reservoir of C. difficile through gastrointestinal carriage in up to 39% of cats and dogs. These findings suggested that direct transmission from household pets, or contamination of the environment by them, may be a factor in the pathogenesis of C. difficile-associated diarrhoea. To investigate this possibility, we examined isolates of C. difficile from humans, pets and the environment by restriction enzyme analysis (REA) and restriction fragment length polymorphism (RFLP) typing using enhanced chemiluminescence. Both REA and RFLP typing methods used Hind III digests of chromosomal DNA. A total of 116 isolates of C. difficile from pets (26), veterinary clinic environmental sites (33), humans (37) and hospital environmental sites (20) was examined. REA was far more discriminatory than RFLP typing and for all isolates there were 34 REA types versus 6 RFLP types. There was good correlation between the REA types found in isolates from pets and from the veterinary clinic environment, and between isolates from humans and from those found in the hospital environment. There was, however, no correlation between REA type of C. difficile found in pets and isolates of human origin. We conclude that there may still be a risk of humans acquiring C. difficile from domestic pets as these findings may be the result of geographical variation.

The use of polyclonal antibodies in differentiating between samples of faecal microflora derived from ten healthy volunteers was assessed. The distribution of FITC-labelled antibodies (of the IgA, IgG and IgM isotype) over 144 morphologically distinct subsets of faecal bacteria (‘morphotypes’) was quantified by means of digital morphometry and quantitative immunofluorescence. Furthermore, a new dataprocessing algorithm was developed which makes objective quantitation of the antibody distributions over the faecal morphotypes possible. The results of this study imply that the antibody binding capacity of faecal morphotypes is a unique characteristic of faecal microflora.

A two-tier miniaturized scheme of eight tests was devised for biotyping strains of Escherichia coli in microwell plates. Primary biotypes were defined by positive and negative reactions in tests for fermentation of raffinose, sorbose, dulcitol and 2-deoxy-D-ribose and for decarboxylation of ornithine when read after specified periods of incubation; subtypes were identified within primary biotypes according to results in secondary tests for rhamnose fermentation, lysine decarboxylation and motility. The method gave reproducible results on different occasions of testing.

Among 100 E. coli strains from various sources, 26 of the 32 possible primary biotypes and 56 full biotypes, as defined by results in both primary and secondary tests, were identified, thus demonstrating a high index of strain discrimination (D = 0·98).

The scheme is recommended as a simple, reliable, inexpensive and efficient method of differentiating strains of E. coli.

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.

A method for typing Haemophilus species is described, based on the analysis of genomic DNA from Haemophilus parainfluenzae. The DNA was extracted by a rapid method and digested with the restriction enzyme BamHI to provide a characteristic ‘fingerprint’. The pattern of fragments in the ranges 1–1·6 kb, 1·6–2 kb and 2–3 kb were used to produce a numerical profile of each isolate. In total 97 isolates were examined; 88 from throat swab material isolated from the 15 members of a British Antarctic Survey base and 9 type strains. Seventy-two of the 88 antarctic isolates were H. parainfluenzae and were found to be very diverse, comprising 41 identifiable strains with up to 5 strains being isolated from a single throat swab sample. There was evidence for both carriage and transmission within the isolated community. The technique provided a highly discriminatory method for characterizing Haemophilus strains which is suitable for epidemiological studies.

Healthy persons were shown to possess circulating antibodies of both IgA, IgG and IgM isotype directed against the bacteria of their faecal microflora, assessed by immunomorphometry. After removal, by absorption, of the fraction of antibodies directed against the autochthonous faecal bacteria or cross-reacting with allogenous faecal bacteria, there were still antibodies left directed against allogenous faecal bacteria of both the IgA, IgG and IgM isotype. However, relatively more antibodies of the IgA isotype appeared to be directed against allogenous bacteria than against indigenous faecal bacteria. Persons who reacted with specific antibodies to many bacteria of their own flora also tended to react specifically to bacteria in the allogenous microflora of the other volunteers. The patterns of antibodies directed to faecal bacteria of different morphologies (morphotypes) were unique for each individual.

Cooling towers have been demonstrated to be amplifiers and disseminators of legionella, the causative organism of Legionnaires' disease. Community outbreaks associated with cooling towers have been reported with several common factors. Small towers (< 300 kW) have predominantly been implicated in outbreaks. Cooling tower-associated outbreaks are most frequent in autumn, and frequently implicated systems have been operated after a period of shutdown.

This paper reports field study data relating system operation to legionella colonization of systems. Operating systems have been shown to be more frequently colonized by legionella than shutdown systems. In some cases operation of systems after periods of shutdown raised legionella concentrations from below detection limits to between 50 and 950 c.f.u./ml within 10 min.

These data and previously reported data relating to biofilm and sediment colonization of the systems, and community outbreaks of Legionnaires' disease, have been used to develop a model explaining the seasonal nature of outbreaks associated with irregularly operated, small cooling tower systems.

Variations in rDNA gene loci in DNA digests of 209 clinical isolates of Serratia marcescens were determined with an Escherichia coli rRNA probe. Forty-one restriction fragment length polymorphism patterns (ribotypes) were identified, based on the size of 4–14 (mean 7·5) hybridization bands. The patterns differed by more than a single band in 98% of pair-wise comparisons. On a subset of 76 isolates, ribotyping proved to be marginally more discriminating than biotyping (discrimination index 0·92 v. 0·89) followed by serotyping (0·87) and bacteriocin typing (0·74). About one-third of isolates belonged to unique ribotypes and only two ribotypes exceeded 5% in frequency (23·0 and 6·4% respectively). A combination of serotype or biotype with ribotyping defined a similar number of strains, although none of the methods alone was sufficiently discriminatory to identify strains. We conclude that due to the accessibility of biotyping and the lack of commercially available antisera for S. marcescens, the biotype and ribotype together provide reliable markers of strain identity.

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.

Anti-pneumococcal polysaccharide antibody (anti-PPS) levels were measured in 153 serum samples collected from children aged between 2 and 47 months living in the highlands of Papua New Guinea (PNG). Fifty-seven of the samples were collected during acute episodes of lower respiratory tract infection (ALRI). Total IgA and IgG increased steadily with age; however, no association was found between the levels of these antibodies and the health status of the child. Total IgM levels showed little relationship to the age of the child but under 12 months of age levels were somewhat higher on average in children with pneumonia. For most of eight pneumococcal serotypes tested, specific IgG levels were found to decline rapidly in the first 6–8 months, reaching a minimum at approximately 12 months of age. Serotype 3 was exceptional in having very low titres in the youngest children. A separate analysis of 24 cord sera suggested that antibodies to this serotype do not usually cross the placenta in PNG. Children with pneumonia tended to have lower levels of specific IgG than healthy controls of the same age. Specific anti-PPS IgA levels were found to increase steadily with age, but were not associated with health status.

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.

Although human visceral leishmaniasis (VL) is a notifiable disease in Italy, there is evidence that the actual number of cases is far higher than that notified. A programme for active surveillance of VL in the 14 Italian endemic regions was launched by the Istituto Superiore di Sanità. We report data collected during a 3-year period of active surveillance in Campania, a south Tyrrhenian region covering 4·5% of the Italian territory. Out of 120 clinically suspected cases referred to medical and diagnostic references centres, there were 52 confirmed VL cases (17·3/year), i.e. 10-fold more than previously notified. Most of the infection sites were in rural areas or peripheral districts of towns in hilly parts of Naples province. An epidemic cluster of 10 cases was identified in a microfocus of Caserta province. The biochemical analysis of 23 Leishmania stocks showed a zymodeme composition indicating Campania as an old and well-established focus of VL. The data obtained emphasize that the present notification system for VL in Italy is inadequate.

The natural habitat of Paracoccidioides brasiliensis, the aetiologic agent of paracoccidioidomycosis, has not been determined. Consequently, the events leading to the acquisition of infection remain controversial. To identify factors associated with infection in endemic areas we conducted a survey in three rural communities in Colombia where we had previously diagnosed paracoccidioidomycosis in children. Permanent residents were surveyed taking into consideration environmental and occupational variables. Skin tests were used to classify subjects as infected or non-infected. Variables found associated with infection were: (i) community A: previous residence around Porce river and agriculture in vegetable gardens; (ii) community C: frequent use of specific water sources; (iii) community V: housekeeping activities, and (iv) total group: age > 25 years and contact with bats. Residents in communities with higher prevalence of infection were older, had more complex residence history, and referred more contact with armadillos than residents of communities with lower infection.