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.

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.

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.

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.

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.

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.

An overview of serologieal and virological studies on poliomyelitis in the Netherlands between two epidemics in 1978 and 1992 is given. Three unvaccinated patients acquired poliomyelitis abroad. In the Netherlands vaccination coverage with quadruple DPT–IPV vaccine is very high. The strong immunogenicity of inactivated poliovirus vaccine was confirmed in a cohort of children, reflected in age-stratified antibody profiles of the population. Adults born in the pre vaccination era appeared in general protected, but 10–25% of persons born between 1930 and 1945 lacked neutralizing antibodies. Revaccination induced a booster type of antibody response in 75–90% of such persons, indicating immunological memory and protection.

Virological studies on adopted children from other countries, patients with indications for viral examination, and river waters showed that the Netherlands was regularly exposed to polio virus (PV), without signs of indigenous transmission. Persons found to carry PV or their close contacts had travelled to a PV endemic country. Most of 557 isolates were vaccine-derived, only 8% were wild type viruses. Despite their presence, up to 1992 the well-known susceptibles for PV in the Netherlands were shielded by the herd immunity of the Dutch population.

Certain health risks have been associated with recreational exposure to faecally polluted water. Canoeing in certain South African waters is considered to be a high risk activity with regard to schistosomiasis. gastroenteritis and possibly hepatitis. In a cross-sectional study, a serosurvey was conducted amongst canoeists to ascertain whether or not they had a higher seroprevalence to hepatitis A virus. Norwalk virus and Schistosoma spp. than non-canoeists. In comparisons between the two groups, a significant association could not be demonstrated between canoeing and antibody response to hepatitis A and Norwalk viruses (P-values for age-adjusted χ2 were 0·083 and 0·219 respectively), but a significant association could be demonstrated between canoeing and the antibody response to Schistosoma spp. (P > 0·001: age-adjusted).

In N'Djamena, capital of Chad, measles vaccination coverage of 12–23-month-old children fell from 61% in 1990 to 15% in 1993. A community survey of measles after an outbreak in 1993 showed that among children < 5 years of age, the mean monthly attack rate was 37 per 1000 (95% CI, 32–43) and the mean case fatality rate was 7·4%. Measles incidence was highest (77/1000/month) in children aged 9–11 months and fell among children > 3 years of age. Incidence rates were high (56/1000/month) among 6–8-month-old children, but only 3 deaths occurred in this age group. Measles vaccine efficacy, estimated by comparing attack rates in unvaccinated and vaccinated children, was 71 % (95% CI, 59–80%). Extrapolation of the results to the city population indicated that an estimated 19000 cases and > 1000 measles-associated deaths occurred in 1993. This preventable morbidity and mortality, in a city where coverage was formerly among the highest in Africa, shows the need for sustained global commitment to preventive health care.

During a large measles outbreak in Quebec City in 1989, two investigations conducted in parallel evaluated the relative risk of measles and measles vaccine effectiveness with respect to age at vaccination. The study was a school-based case-control study including 563 cases and 1126 classmate controls. The second was a cohort study of the siblings of school cases including 493 siblings aged between 1 and 19 years. The relative risks (RR) of measles were similar in both settings and the trend towards increased vaccine efficacy with increasing age at vaccination was highly significant (P < 0·001). Vaccine efficacy rose from 85% in children vaccinated at 12 months of age to ≥ 94% in those vaccinated at 15 months and older. Even for children vaccinated at or after 18 months of age. the RR of measles was reduced when compared with children vaccinated between 15 and 17 months of age (RR 0·61. CI 95% 0·33–1·15). Small changes in the timing of initial measles vaccination can have a major impact on vaccine efficacy.

Mathematical models are potentially useful tools to aid in the design of control programmes for parasitic diseases. In this paper, a fully age structured epidemiological model of human schistosomiasis is developed and parameterized, and used to predict trends in infection prevalence, intensity and prevalence of heavy infections over age and time during several rounds of mass and age targeted treatment. The model is validated against data from a Schistosoma mansoni control programme in Kenya.

The susceptibility of laboratory reared Zimbabwean Amblyomma hebraeum and A. variegatum ticks to infection with geographically distinct Cowdria ruminantium strains was investigated by feeding both species simultaneously on individual sheep infected with one of the four strains (Crystal Springs [Zimbabwe], Ball 3 [South Africa]. Gardel [Guadeloupe] and Nigeria [Nigeria]). A. hebraeum ticks demonstrated a high susceptibility to infection with all four C. ruminantium strains. In comparison. A. variegatum were less susceptible to infection with the Crystal Springs and Ball 3 strains (P < 0·001), but showed a similar susceptibility to the Gardel and Nigeria strains. The differences in susceptibility of A. variegatum to infection with the four strains of C. ruminantium correlated with the origin of these strains. The consistently higher susceptibility of A. hebraeum ticks to infection with geographically different C. ruminantium strains may be one explanation for the observation that heartwater is a more serious problem where A. hebraeiun is the vector of the disease.

The suitability of serological surveys of roe deer (Capreolus capreolus) in determining the spread of tick-borne encephalitis virus (TBEV) was tested in a south German area with a low risk of TBEV infection to humans. Sera obtained from 192 hunted roe were screened by an haemagglutination–inhibition test (HAI) and in an ELISA developed in our laboratory. Those found positive were tested in a neutralization test (NT). Fifty (26·0%) sera reacted positive by ELISA and 43 (86·0%) of these were confirmed by HAI or NT. Forty-seven (24·5%) samples were positive by HAI, 44 (93·6%) of which were also positive in NT or ELISA. Only insignificant increase of the antibody prevalence with age (P = 0·17 for HAI antibodies) suggests that most infections occur at an early age in scattered natural foci. The antibody prevalence in females was lower than in males (OR = 0·63; P = 0·02 for HAI antibodies). In determining the distribution of seropositive roe we increased the sample size to 235 sera. No antibodies were detected in 56 (23·8%) sera collected in the eastern third of the county. The areas of high antibody prevalence in roe match those in which humans have been infected. We conclude that serosurveys of roe deer are useful in marking out areas in which humans face the risk of infection, provided that an adequate number of sera, preferably from males, is available.

Methods of transmission of Jembrana disease, an acute and severe disease of Bali cattle (Bos javanicus) caused by a recently-identified bovine lentivirus known as Jembrana disease virus, are described. During the acute disease virus can be detected in saliva and milk. There is evidence of direct transmission from acutely affected animals in close contact with susceptible cattle, possibly by virus in these secretions infecting cattle by the conjunctiva!, intranasal or oral routes, by which it was possible to infect cattle experimentally. During the acute disease the titre of infectious virus in blood is high, about 108 50% cattle infectious units (ID50)/ml, and it is probable that the virus is also transmitted mechanically by haematophagous arthropods. Recovered cattle are also a potential but probably infrequent source of infection; recovered cattle are persistently viraemic but the titre of infectious virus in blood decreases to about 101 ID50/ml by 60 days after recovery from the acute disease, and virus cannot be detected in secretions.