Campylobacter species such as C. jejuni and C. coli are recognized as major causes of acute gastroenteritis world-wide. Although C. jejuni and C. coli are usually non-pathogenic in birds and animals, they cause enteric disease in humans and the source of infection is often the consumption of contaminated foodstuffs. In this paper we report on the development and use of a multiplex PCR of C. jejuni genomic DNA which yielded a PCR product with a unique polymorphic site that can be used to quickly and accurately identify and group C. jejuni isolates from any source including DNA, cell culture, skin washings and faecal samples. The test is simple and sensitive and can detect purified DNA from a single bacterium 102 cells from crude lysates, 103 cells in seeded faeces and 120 cells/ml of washing of 1 cm2 skin fragments of chicken skin.

An unusually high incidence of Vibrio cholerae O1 infection was observed in southern Thailand between late December 1997 and March 1998. Fifty-seven V. cholerae O1 strains were isolated in five provinces during this epidemic and were examined. They were El Tor Ogawa strains exhibiting similar antibiograms. All strains were resistant to tetracycline, which had not been reported in Thailand since 1993. The ribotypes, hybridization patterns with ctx and zot gene probes, arbitrarily primed PCR profiles, and pulsed-field gel electrophoresis profiles of the representative strains were compared with the clinical strains isolated from patients in India and Bangladesh in 1997 and 1998 and from international travellers originating from various Asian countries during the 1992–8 period. All southern Thailand strains and the 1998 international traveller strain of Thai origin showed indistinguishable genetic fingerprinting patterns that were distinct from those of other test strains. The results suggest that a tetracycline-resistant clone newly emerged in late December 1997 caused the large epidemic in southern Thailand and that the variants with a slightly different antibiogram appeared during the course of the spreading epidemic.

The phenotypic and genotypic profiles of the V. cholerae strains causing the Mozambican 1997–8 epidemic were characterized to provide a reference for comparison with other epidemic strains. A total of 75 strains of V. cholerae O1 isolated in different provinces, were analysed. Strains were characterized by PCR for detecting toxin genes (ctxA, zot and ace), virulence associated genes (tcpA, nanH, hlyA and toxR) and ERIC sequences. All V. cholerae strains were serotype O1, Ogawa, biotype El Tor. MIC testing showed a high proportion of strains multi-resistant to drugs (100% to cotrimoxazole and 52% to tetracycline) and susceptibility to ciprofloxacin. The isolates contained two intact copies of the CTX genetic element and all other genes tested. PCR of restricted DNA revealed two ERIC types: the first in provincial isolates, also predominant in other African epidemic strains, and the second in Maputo isolates (the national capital).

Nucleotide sequences of Shiga toxin (Stx) genes in STEC from various origins were determined and characterized by phylogenetic analysis based on Shiga toxin (Stx) with those deposited in GenBank. The phylogenetic trees placed Stx1 and Stx2 into two and five groups respectively, and indicated that Stx1 in sheep-origin STEC were placed into a different group from those in other STEC, and that Stx2 of deer-origin STEC also belonged to the unique group and appeared to be distantly related to human-origin STEC. On the other hand, Stx of STEC isolated from cattle, seagulls and flies were closely related to those of human-origin STEC. Such a diversity of Stx suggested that STEC might be widely disseminated in many animal species, and be dependent on their host species or their habitat. In addition, the active sites in both toxins were compared; the active sites in both subunits of Stx in all the animal-origin STEC were identical to those in human-origin STEC, suggesting that all the toxin of STEC from animals might be also cytotoxic, and therefore, such animal-origin STEC might have potential pathogenicity for humans.

The sources and transmission routes of sporadic Yersinia enterocolitica bioserotype 4/O:3 infections in Finland were studied. A total of 212 human strains were compared with 334 non-human strains, including 163 strains from pig slaughterhouses, 164 strains from retail outlets and 7 strains from pet animals. All strains were characterized using pulsed-field gel electrophoresis (PFGE) with NotI enzyme. When the 194 human and 287 non-human strains of 22 identical NotI profiles were further characterized with ApaI and XhoI enzymes, 126 genotypes (DI = 0·94) were distinguished. Of all 212 human strains, 80% were genetically indistinguishable from the strains found in samples of pig origin when characterized with the three enzymes. A major contamination source of sporadic Y. enterocolitica 4/O:3 infections was revealed to be edible pig offal: 71% of the human strains were indistinguishable from the strains isolated from tongues, livers, kidneys and hearts of pigs. These results reveal that in Finland contaminated pig offal is an important vehicle in the transmission of Y. enterocolitica bioserotype 4/O:3 from slaughterhouses to humans.

A cross-sectional survey was conducted to determine prevalence and assess risk factors for carriage of antibiotic resistant strains of Streptococcus pneumoniae in healthy school children in Hong Kong. Throat swabs were collected from 1455 subjects and written questionnaires providing demographic data and medical history were completed by parents. The overall carriage rate of Streptococcus pneumoniae was 3·5%, of which 49% were penicillin resistant. High levels of resistance to tetracycline (73%), erythromycin (52%), trimethoprim (66%) and ciprofloxacin (57%) were observed. Carriage was associated with presence of a younger sibling (OR = 1·79) and use of antibiotics (OR = 2·31). High use of day care and small size of housing units did not result in a high rate of carriage. The low rate of carriage may be linked to high use of antibiotics, geographical factors or ethnicity. High rates of antibiotic resistance reflect heavy use of antibiotics by general practitioners.

We describe the isolation and molecular characterization of methicillin-resistant coagulase-negative staphylococci (MRCNS) from the nasal flora of healthy humans from three institutions located in Rio de Janeiro City. Swabs were obtained from the nares of students attending a non-residential public school and adults from two military quarters. Isolates of staphylococci were tested for the presence of the mecA gene by hybridization with a specific probe. S. epidermidis was the most frequent MRCNS (38 of the total 45 CNS isolated). Twenty-five percent of nasal staphylococcal carriers studied were colonized with MRCNS. Pulsed-field gel electrophoresis (PFGE) of SmaI-digested genomic DNA was carried out to study the clonality of the methicillin-resistant S. epidermidis (MRSE) isolates. In addition to cross-colonization among individuals belonging to the same institution, familial cross-colonization appeared to contribute to the spread of the methicillin-resistant isolates among two inter-communicable institutions. Indeed, the wide genomic diversity among the MRSE flora suggests that the spread of the mecA gene among these isolates might also have occurred via horizontal transmission. Despite the limited number of institutions analysed, it is reasonable to conclude that our data do not represent a situation unique to the three organizations but may reflect other communities in Rio with respect to transmission of MRCNS.

The incidence of pertussis requiring hospitalization in children younger than 16 years was estimated by the use of an active surveillance-system. Of special interest were differences between West and East Germany following different vaccination strategies before reunification. In 1997 and 1998, 754 pertussis cases required a total of 11151 hospital inpatient days. The incidence of hospitalized pertussis was 2·68/100000 person years and this was significantly higher in East than in West Germany. In East Germany an unusually high percentage of hospitalized cases was found in children aged 6–15 years (45% versus 13% in West Germany). The difference between the regions may be due either to a different perception of the disease or to an increased immunity induced by prior disease or vaccination. In East Germany, pertussis was rare until reunification but it has increased significantly since then. Older children may thus represent a population at risk of pertussis having not had previous exposure to pertussis antigens.

In the Netherlands a strong epidemic outbreak of pertussis took place in 1996–7. Here we investigate the possible causes of the epidemic, using an age-structured epidemic model. Motivated by the observation that during the epidemic the number of cases in vaccinated children had increased considerably compared to the preceding period, we focus on two vaccination related changes. First, we consider the possibility that the potency of the vaccine decreased so that it confers protection for a shorter period of time in newly vaccinated children. Second, we consider the possibility that at a certain point in time the duration of protection after vaccination decreased for all individuals. This may be the case if the pathogen population changed such that the current vaccine confers less protection. A comparison of the observed and simulated age-distribution of infections indicates that the second scenario is more in line with the observed pattern of the 1996–7 epidemic. We discuss the implications of this conclusion for B. pertussis circulation, and for the design of vaccination programmes in the face of a polymorphic B. pertussis population that may adapt itself to vaccination.

The purpose of the study was to describe the results of follow up of human contacts of bovine tuberculosis. The bovine tuberculosis cases occurred on farms in North Staffordshire between 1993 and 1997. A total of 162 people were identified as having close contact with cattle diagnosed as having bovine tuberculosis, or who had drunk unpasteurized milk from a herd with bovine tuberculosis. A retrospective review of chest clinic notes was performed. One hundred and thirty-eight people attended for follow up, and Heaf test results, necessity for chest X-ray and further clinical follow-up are described. No case of human Mycobacterium bovis infection was identified. It is suggested that follow-up of human contacts is limited to those with close contact with herds who have bovine tuberculosis and cattle with visible pulmonary lesions or evidence of udder infection. Children on the farms with affected cattle should also be offered BCG in advance of the routine school's programme.

Individuals developing an HIV seroconversion illness may experience rapid disease progression. Information on seroconversion illness is rarely collected in most cohort studies; thus the aim of this study was to assess the value of the HIV test interval (the time between last negative and first positive HIV tests) as a proxy for seroconversion illness. Among 8229 seroconverters, test intervals ranged from 0–5282 days, and varied by gender, risk group, age and calendar year of seroconversion. Those with intervals [les ] 31 days had an increased hazard of AIDS (RH 1·42, P = 0·07), which was reduced slightly after adjusting for baseline factors, calendar year of follow-up, treatment and the declining CD4 count, but there was no effect on survival. Thus, it appears that if information on acute seroconversion illness is not available, then analyses of progression to AIDS in seroconverter studies could use a short test interval as a proxy measure.

In order to assess the feasibility of environmental poliovirus surveillance, known amounts of poliovirus type 1, strain Sabin, were flushed into the sewage network of Helsinki. Grab specimens collected at a remote downstream location and concentrated about a 100-fold revealed infectious poliovirus on four successive days in all three separate experiments. As for concentration, a simple two-phase separation method was found to be at least as useful as a several-fold more resource-demanding polyethylene glycol (PEG) precipitation method. Recovery of the introduced virus was remarkably high (more than 10%). Using the current system, it might be possible to detect poliovirus circulation in a population of 700 000 people by examining a single 400 ml sewage specimen, if 1 out of 10000 inhabitants were excreting the virus. It is concluded that environmental surveillance is a sensitive approach to monitor silent poliovirus circulation in populations served by a sewage network.

Two epidemics of acute, watery diarrhoea in villages in North Arcot district, India, were investigated. The attack rates were 10·03 and 15·53 per 100 population, the median duration was 5 days and enteric pathogens were present in 56·8% and 60·3% of specimens from the two villages, but no predominant pathogen was identified. Examination of stools from a 20% age-stratified random sample of the population of one of the villages after the epidemic found 22·9% of asymptomatic subjects excreted bacterial enteric pathogens. Despite the high background of enteric pathogen carriage, the isolation rates for shigellae, enteropathogenic Escherichia coli and Shiga-toxin producing E. coli were significantly higher (P < 0·001, P < 0·02, P < 0·05) during the epidemic. The epidemics may have been caused by faecal contamination of well water following rain. Point-of-use techniques for water disinfection may be most effective for preventing such outbreaks, but further research into the development of appropriate technology is required.

We compared the relative risks of mother-to-child transmission of Toxoplasma gondii and clinical manifestations due to congenital toxoplasmosis associated with intensive prenatal treatment in Lyon and Austria, short term treatment in 51% of Dutch women, and no treatment in Danish women. For each cohort, relative risks were standardized for gestation at seroconversion. In total, 856 mother–child pairs were studied: 549 in Lyon, 133 in Austria, 123 in Denmark and 51 in The Netherlands. The relative risk for mother-to-child transmission compared to Lyon was 1·24 (95% CI: 0·88, 1·59) in Austria; 0·59 (0·41, 0·81) in Denmark; and 0·65 (0·37, 1·01) in The Netherlands. Relative risks for clinical manifestations compared with Lyon (adjusted for follow-up to age 3 years) were: Austria 0·19 (0·04, 0·51); Denmark 0·60 (0·13, 1·08); and The Netherlands 1·46 (0·51, 2·72). There was no clear evidence that the risk of transmission or of clinical manifestations was lowest in centres with the most intensive prenatal treatment.

To estimate the burden of disease due to congenital toxoplasmosis in Sweden the incidence of primary infections during pregnancy and birth prevalence of congenital toxoplasmosis in 40978 children born in two regions in Sweden was determined. Women possibly infected during pregnancy were identified based on: 1, detection of specific IgG based on neonatal screening of the phenylketonuria (PKU) card blood spot followed by retrospective testing of stored prenatal samples to detect women who acquired infection during pregnancy and follow up of their children to 12 months; 2, detection of specific IgM on the PKU blood spot.

The birth prevalence of congenital toxoplasmosis was 0·73/10000 (95% CI 0·15–2·14) (3/40978).

The incidence of primary infection during pregnancy was 5·1/10000 (95% CI 2·6–8·9) susceptible pregnant women. The seroprevalence in the southern part was 25·7% and in the Stockholm area 14·0%.

The incidence of infection during pregnancy was low, as the birth prevalence of congenital toxoplasmosis. Neonatal screening warrants consideration in view of the low cost and feasibility.

The aim of this study was to describe the occurrence of shiga toxic genes (stx) in Norwegian sheep herds, and to identify herd management factors related to the occurrence of stx in herds. Faecal samples from 124 sheep-herds were collected at abattoirs in 1998. Pooled samples from lambs and from ewes were screened for stx by a PCR method directly on faeces. Of the 124 herds, 61 were positive for stx, giving an overall herd-prevalence of 49%. Twenty-one of the 61 positive herds were positive both in lamb and ewe samples, 24 only in lamb samples and 16 only in ewe samples. There was no difference in prevalence between regions. From the 21 herds positive both in lamb and ewe samples, stx encoding E. coli were detected in 18 herds using hydrophobic grid membrane filters and subsequent colony hybridization. Information about management factors was collected by telephone interviews. Having cattle at the same farm turned out to be a possible risk factor, with an Odds Ratio of 9·9 (CI 1·2 → ∞).

Nine isolates from pigs persistently infected with a recent Italian isolate of swine vesicular disease (SVD) virus, ITL/9/93, were collected sequentially over 121 days and were characterized antigenically and biochemically. There was an accumulation of amino acid (aa) substitutions in the capsid proteins throughout the carrier state that could be correlated with alterations in antigenicity in virus isolates collected late stage in infection. The aa substitutions detected mainly occurred in VP1 and antigenic changes were detected in late isolates both at antigenic site 1, resulting in loss of binding of Mab 4GO7, and at a closely located site which has not yet been named, recognized by Mab C29. In further experiments groups of pigs were exposed to a range of SVD viruses, but no virus was isolated beyond 16 days post infection (dpi) nor viral RNA detected beyond 42 dpi. Attempts to transfer infection to sentinel pigs introduced some time after initial infection of the original pigs were largely unsuccessful. The carrier state was established in only one out of five experimental infections of pigs with SVD virus and can therefore be considered a rare sequel to infection with SVD virus and is of limited significance in the epidemiology of the disease.

The prevalence of three waterborne zoonotic pathogens (Campylobacter sp., Giardia sp. and Cryptosporidium parvum) in rectal faecal samples from a random sample of adult beef cattle was determined. Management factors that may be associated with shedding of these organisms were examined. For Campylobacter sp. prevalence was 5·0%, and the number of females on the farm was positively associated with the proportion that tested positive. For Giardia sp. prevalence was 6·5%, and none of the management factors examined was significantly associated with the proportion in a herd testing positive. C. parvum was identified in 1·1% of samples. The length of calving season and whether any procedures were performed on the calves in the first 2 days of life were positively associated with the proportion that tested positive. We conclude that this sample of adult beef cattle represent a relatively limited threat to water supplies and subsequent disease transmission to humans from these pathogens.

The epidemiology and transmission dynamics of sheep scrapie is as yet poorly understood. Here we present a theoretical analysis of the transmission dynamics within a sheep flock, concentrating on how persistence properties depend on transmission scenario and flock size. Patterns of disease persistence and extinction are studied analytically using branching-process approximations and numerically using stochastic model simulations. For a given basic reproduction number, disease extinction is most likely when late-stage infected animals are responsible for most of the transmission. This effect can be understood in terms of aggregation in the distribution of the number of secondary infections arising from a single primary infection. The presence of an environmental reservoir reduces the probability of extinction.

We used spoligotyping to study 500 randomly selected pretreatment Mycobacterium tuberculosis (MTB) strains isolated in Hong Kong during the 2 year period 1998–9. It was found that amongst all MTB strains studied, the ‘Beijing’ genotype strains were highly prevalent in our geographic area, representing about 70% of the isolates. Unlike previous observations in Vietnam, no significant associations were found either between ‘Beijing’ genotype strains and all other anti-tuberculosis drug resistance phenotypes, or with particular patients' age groups, except for a weak association with isoniazid susceptibility. Eighteen of these strains exhibited spoligotype patterns that were similar but not identical to the ‘Beijing’ specific pattern. This is the first geographical area where genetic diversity among ‘Beijing’ genotype of MTB strains has been observed on this scale.