There is very little human disease associated with enterohaemorrhagic Escherichia coli O157 in Australia even though these organisms are present in the animal population. A group of Australian isolates of E. coli O157:H7 and O157:H- from human and animal sources were tested for the presence of virulence markers and compared by XbaI DNA macrorestriction analysis using pulsed-field gel electrophoresis (PFGE). Each of 102 isolates tested contained the gene eae which encodes the E. coli attaching and effacing factor and all but one carried the enterohaemolysin gene, ehxA, found on the EHEC plasmid. The most common Shiga toxin gene carried was stx2c, either alone (16%) or in combination with stx1 (74%) or stx2 (3%). PFGE grouped the isolates based on H serotype and some clusters were source specific. Australian E. coli O157:H7 and H- isolates from human, animal and meat sources carry all the virulence markers associated with EHEC disease in humans therefore other factors must be responsible for the low rates of human infection in Australia.

Multidrug-resistant Salmonella enterica subsp. enterica serotype Typhimurium DT104L was first reported in Singapore from mid-July to mid-October 2000. Salmonella strains isolated from clinical laboratories were submitted to a reference laboratory for serotyping, phage-typing and pulsed-field gel electrophoresis (PFGE) using XbaI restriction endonuclease. An epidemiological investigation was conducted to determine the source of infection and mode of transmission using a structured questionnaire. A total of 33 cases involving mainly infants and toddlers were detected in the 3-month long outbreak. The outbreak strain was of the R-type ACGSTSu, i.e. resistant to ampicillin, chloramphenicol, gentamicin, streptomycin, tetracycline and sulphonamide. PFGE showed all isolates had an indistinguishable pattern, indicating a common source of infection. Consumption of imported dried anchovy was found to be the vehicle of transmission after adjusting for all confounding variables in the case-control study using stepwise logistic regression (OR 25·6; 95% CI 3·9–167·9; P = 0·001). Imported dried seafood should be properly processed, packed, labelled, and thoroughly cooked to prevent transmission of multidrug-resistant S. Typhimurium.

Although campylobacter has been the most commonly recognized bacterial cause of gastro-intestinal infection in England and Wales since 1981, there are few reported campylobacter outbreaks. Of the 2374 general outbreaks of infectious intestinal disease reported to CDSC between 1995 and 1999, for which an aetiological agent was identified, campylobacter accounted for only 50 (2%). Foodborne transmission was identified in 35 outbreaks and the majority took place in commercial catering establishments; waterborne transmission was responsible for a further four outbreaks. Isolates of Campylobacter jejuni were referred for typing from 25 outbreaks. In 13 outbreaks all isolates were the same subtype, as defined by serotype and phage type, while in the remainder more than one campylobacter subtype was involved.

In the years 1999–2000, there was an increase in the incidence of meningococcal disease in Victoria, largely caused by Neisseria meningitidis serogroup C. This change was associated with a shift in age distribution of cases, with relatively more disease appearing in the 15–29 year age group, and with 40/58 serogroup C isolates in 2000 exhibiting a new macrorestriction pattern (pattern A). Thirty-four of 52 pattern A isolates tested displayed the novel phenotype C:2a:P1.4, and were consistently porA VR type P1.7-2,4 by DNA sequencing. Nine of 10 representative pattern A isolates analysed displayed a housekeeping gene allele profile (ST-11) that is characteristic of the electrophoretic type (ET)-15 variant that has caused outbreaks in Canada, the Czech Republic and Greece. Meningococci belonging to the ST-11 complex that were isolated in Victoria prior to 1999 did not display either restriction pattern A or PorA VR type P1.7-2,4.

Some years ago, an increase in the number of sporadic cases and outbreaks of salmonellosis due to S. enterica serovar Bovismorbificans was observed in several European countries including Finland, Sweden, England/Wales, Austria, and Germany. In order to understand the recent spread of this serovar and to trace the route of infection back to its source, it was considered necessary to subtype S. Bovismorbificans isolates. Using phage typing (newly described here) and molecular fingerprinting (PFGE-pattern, plasmid profiles and ribotype) the isolates of European origin could be subtyped and compared to S. Bovismorbificans isolates that originated in overseas countries such as Australia, Thailand, India, etc. where this serovar was isolated more frequently. Significant clonal diversity was identified but some of the clonal types of S. Bovismorbificans dominated the epidemics and single cases in Europe as well as in overseas countries. The clonal identity among these isolates indicates an international distribution, new sources of infection, and highlights the urgent requirement for standardized laboratory based surveillance networks (e.g. Enter-Net). Moreover, it is suggested that strains of S. Bovismorbificans will continue to be of concern in public health and that phage typing together with PFGE typing can be applied as reliable and rapid tools for their future monitoring.

In an outbreak of gastroenteritis on 23 July 1996, in Osaka, Japan, 54 of 91 persons who had attended a meeting the previous day became ill. Escherichia coli O166[ratio ]H15 was isolated from stool specimens of patients (29/33, 88%). Laboratory tests for other bacterial pathogens and viruses were negative. The E. coli O166 organisms did not adhere to HEp-2 cells in a localized, diffuse, or enteroaggregative manner. The organisms did not express known enterotoxigenic E. coli (ETEC) colonization factors. In polymerase chain reaction tests, the bacteria did not have coding genes for shigatoxin of enterohemorrhagic E. coli (EHEC), heat-labile, or heat-stable enterotoxin of ETEC, attachment and effacement (eaeA) of EPEC, or invasion (invE) of enteroinvasive E. coli (EIEC). Consequently, they could not be assigned to any of the recognized diarrhoeagenic groups of E. coli: EPEC, ETEC, EHEC, EIEC, enteroaggregative E. coli (EAggEC), or diffusely adhering E. coli. However, the organisms possessed the EAggEC heat-stable enterotoxin (EAST1) gene. To our knowledge, this is the first report of an outbreak caused by E. coli that did not have well-characterized virulence genes other than EAST1. The isolates showed the same DNA banding pattern in pulsed-field gel electrophoresis after digestion with the restriction enzymes XbaI or NotI. Three O166[ratio ]H15 strains isolated from two sporadic cases and another outbreak during 1997–8 were distinct, indicating that multiple clones have spread already. We propose that diarrhoeal specimens should be examined for E. coli possessing the EAST1 gene.

On 6 May 2000, a staphylococcal food poisoning outbreak occurred at a high school, affecting 10 of the 356 students who attended the breakfast. Twenty-seven Staphylococcus aureus isolates, producing enterotoxin A (SEA), SEB-, or non-SEA-E, were recovered from 7 patients, 2 food handlers and left-overs. To investigate the outbreak, we genotyped the isolates by using pulsed-field gel electrophoresis (PFGE) and three PCR-based techniques: inter-IS256 PCR typing, protein A gene (spa) typing, and coagulase gene restriction profile (CRP) analysis. Our results show that PFGE was the most discriminatory technique, whereas the three PCR-based techniques were insufficient in the discriminatory power to distinguish the S. aureus isolates from the outbreak. Based on the enterotoxin-producing types and the results of genotyping, three distinct types of strains (A1111, B2221 and N3221) were designated. Both the A1111 and B2221 strains were found in the specimens from the patients and a hand lesion of a food handler, suggesting that the source of contamination for the outbreak was most likely originated from a food handler.

The molecular epidemiology of a representative collection of sporadic foreign and domestically acquired Salmonella Typhimurium (S. Typhimurium) isolates from Norwegian patients in 1996–9 was studied by numerical analysis of pulsed-field gel electrophoresis (PFGE) profiles. Three subclusters (E5, F1 and G1) comprised 47% of the 102 sporadic isolates investigated and 45% of the domestically acquired isolates fell in subclusters E5 and F1. Distinct seasonal and geographic variations were evident for these strains which have been responsible for both local outbreaks (E5) and a national epidemic (F1) where salmonella-infected hedgehogs and birds constituted the suggested primary source of infection. Subcluster G1 was dominated by imported multi-resistant definitive type (DT) 104 isolates. All multi-resistant isolates contained integron-associated gene cassette-structures. This study presents valuable information on the relative significance, geographic distribution and antibiotic resistance features of distinct S. Typhimurium clones causing human salmonellosis among Norwegians.

An epidemiological study was carried out to find out the aetiological agent for diarrhoeal disorders in the cyclone and flood affected areas of Orissa, India. Rectal swabs collected from 107 hospitalized diarrhoea patients were bacteriologically analysed to isolate and identify the various enteropathogens. Detection of toxic genes among E. coli and V. cholerae was carried out by polymerase chain reaction (PCR) assay. Of the 107 rectal swabs analysed, 72·3% were positive for V. cholerae O1 Ogawa, 7·2% for V. cholerae O139, 1·2% for E. coli (EAggEC) and 1·2% for Shigella flexneri type 6. Using multiplex PCR assay it was found that all V. cholerae isolates were ctxA positive and El Tor biotype. Strains of V. cholerae O1 were observed to be resistant to nalidixic acid, furazolidone, streptomycin, co-trimoxazole and ampicillin. Except for nalidixic acid, the resistance pattern for O139 was identical to that of O1 strains. Representative strains of V. cholerae were further characterized by randomly amplified polymorphic DNA (RAPD) analysis and ribotyping. Both O1 and O139 V. cholerae strains exhibited the R3 pattern of ribotype and belonged to a similar pattern of RAPD compared with that of Calcutta strains. Early bacteriological and epidemiological investigations have revealed the dominance of V. cholerae O1 among the hospitalized patients in cyclone affected areas of Orissa. Drinking water scarcity and poor sanitation were thought to be responsible for these diarrhoeal outbreaks. Timely reporting and implementation of appropriate control measures could contain a vital epidemic in this area.

We reviewed laboratory data on non-invasive pneumococcal isolates reported from all diagnostic laboratories in Scotland during the period 1988–99. Of 4491 isolates from hospitalized patients, 654 (64·7%) were from sputum, 79 (7·8%) from the nasopharynx and 278 (27·5%) from other superficial sites. The serogroups included in the 23-valent polysaccharide vaccine caused 96–9% of all non-invasive disease in all age groups. The 7-, 9-, and 11-valent conjugated vaccine serogroups were responsible for 87–94%, 85–93%, 74–81% and 75–84% of non-invasive disease respectively in age groups <2 years, [les ]5 years, [ges ]65 years and all ages. The coverage of non-susceptible penicillin and erythromycin non-invasive isolates was >99% and >95% with the 23-valent polysaccharide and 7–11-valent conjugate vaccines respectively. The eight most common serogroups were 23, 9, 6, 19, 14, 3, 15 and 11 (in descending order). The serogroups associated with antimicrobial resistance in non-invasive disease were similar to those found in invasive disease. The finding of a similar serogroup distribution in both invasive and non-invasive disease (regardless of the site of clinical isolate), is consistent with serogroups colonizing non-sterile sites and having the potential to invade. The availability of conjugated vaccines reinforces the importance of systematic surveillance to determine accurately and regularly the coverage of pneumococcal serogroups and types causing both invasive and non-invasive disease.

A review of the epidemiology of invasive pneumococcal disease in Scotland was carried out using data from laboratory-based systems during the period 1988–99. This comprised 5456 (90·8%) isolates of Streptococcus pneumoniae from blood, 467 (7·8%) from cerebrospinal fluid (CSF) and 84 (1·4%) from other sterile sites. The mean annual incidence of invasive disease was 9·8/105 population (9·0/105 for bacteraemia and 0·8/105 for meningitis). Invasive disease was highest in children <2 years of age and in the elderly [ges ]65 years (44·9/105 and 28·4/105 population in these age groups respectively). The highest incidence of pneumococcal meningitis, 11·8/105 persons occurred in children <2 years of age. Males had a higher incidence of pneumococcal bacteraemia and meningitis than females (male[ratio ]female = 1·2[ratio ]1 for bacteraemia (RR = 1·17, 95% CI 1·11, 1·24) and 1·5[ratio ]1 for meningitis (RR = 1·41, 95% CI 1·18, 1·70)). Pneumococcal disease was highest in winter periods and coincided with influenza activity. The proportion of penicillin and erythromycin non-susceptible isolates increased from 4·2% in 1992 to 12·6% in 1999 and from 5·6% in 1994 to 16·3% in 1999 respectively. Our data confirm the substantial and increasing disease burden from pneumococcal disease and rise in prevalence of antibiotic non-susceptibility among pneumococci in Scotland. Continued surveillance of groups at increased risk for pneumococcal disease and the antibiotic susceptibility and serotype distribution of isolates are important to develop appropriate policies for the prevention of pneumococcal disease in Scotland.

In all temperate countries campylobacter infection in humans follows a striking seasonal pattern, but little attention has been given to exploring the epidemiological explanations. In order to better characterize the seasonal patterns, data from nine European countries and New Zealand have been examined. Several European countries with weekly data available showed remarkably consistent seasonal patterns from year to year, with peaks in week 22 in Wales, week 26 in Scotland, week 32 in Denmark, week 30 in Finland and week 33 in Sweden. In Europe, the seasonal peak was most prominent in Finland and least prominent in Scotland and Austria. In New Zealand the seasonality was less consistent since the peak was more prolonged. Possible explanations for the seasonal peaks are discussed. Research into the causes of campylobacter seasonality should help considerably in elucidating the sources of human infection.

Streptococcus pyogenes (group A streptococcus) strains may express several distinct fibronectin-binding proteins (FBPs) which are considered as major streptococcal adhesins. Of the FBPs, SfbI was shown in vitro to promote internalization of the bacterium into host cells and has been implicated in persistence. In the tropical Northern Territory, where group A streptococcal infection is common, multiple genotypes of the organism were found among isolates from invasive disease cases and no dominant strains were observed. To determine whether any FBPs is associated with invasive disease propensity of S. pyogenes, we have screened streptococcal isolates from bacteraemic and necrotizing fasciitis patients and isolates from uncomplicated infections for genetic endowment of 4 FBPs. No difference was observed in the distribution of sfbII, fbp54 and sfbI between the blood isolates and isolates from uncomplicated infection. We conclude that the presence of sfbI does not appear to promote invasive diseases, despite its association with persistence. We also show a higher proportion of group A streptococcus strains isolated from invasive disease cases possess prtFII when compared to strains isolated from non-invasive disease cases. We suggest that S. pyogenes may recruit different FBPs for different purposes.

We examined the serological characteristics of 774 invasive meningococcal isolates collected through an active laboratory-based surveillance system in Scotland from 1994 to 1999. Of these, 72–73% of isolates were tested for susceptibility to several antimicrobial agents. Meningococci with high-level resistance to sulphadiazine had a prevalence of 10% and incidence of 0·22 per 100000 population. High-level resistance to penicillin and other antibiotics was not detected. The prevalence of moderate penicillin resistant meningococci was 8·3%. There was no increase in moderate penicillin resistant meningococcal isolates during the study period, but there were temporal and geographic variations. The estimated incidence of moderate penicillin resistant meningococci was 0·15 per 100000 population. High and low incidence of moderate penicillin resistant meningococci appeared to correlate with the number of doses of penicillin prescribed in some geographic locations. The majority of moderate penicillin resistant isolates belonged to serogroups B (52·2%) and C (39·2%). However, the prevalence of moderate penicillin resistance in serogroup W135 was substantially higher (51·7%) than serogroups B (7·8%) and C (7·6%). Serogroup W135 accounted for a higher proportion of moderate penicillin resistance (8·7%) than disease (1%). There was no predominant penicillin resistant serotype/subtype within any serogroup. Constant surveillance is necessary to monitor the emergence and spread of resistance and to guide appropriate public health interventions in preventing drug resistant meningococci.

The objective was to describe the current epidemiology and trends in New Zealand human leptospirosis, using descriptive epidemiology of laboratory surveillance and disease notification data, 1990–8. The annual incidence of human leptospirosis in New Zealand 1990–8 was 4·4 per 100000. Incidence was highest among meat processing workers (163·5/100000), livestock farm workers (91·7), and forestry-related workers (24·1). The most commonly detected serovars were Leptospira borgpetersenii serovar (sv.) hardjo (hardjobovis) (46·1%), L. interrogans sv. pomona (24·4%) and L. borgpetersenii sv. ballum (11·9%). The annual incidence of leptospirosis declined from 5·7/100000 in 1990–2 to 2·9/100000 in 1996–8. Incidence of L. borgpetersenii sv. hardjo and L. interrogans sv. pomona infection declined, while incidence of L. borgpetersenii sv. ballum infection increased. The incidence of human leptospirosis in New Zealand remains high for a temperate developed country. Increasing L. borgpetersenii sv. ballum case numbers suggest changing transmission patterns via direct or indirect exposure to contaminated surface water. Targeted and evaluated disease control programmes should be renewed.

MLST typing of Neisseria meningitidis directly from clinical material was introduced in the National Reference Laboratory for Meningococcal Infections in Prague. Four cerebrospinal fluid samples were obtained from patients with suspected meningococcal invasive disease. In all samples, all classical laboratory methods gave negative results and the only positive method was PCR, which revealed Neisseria meningitidis group C (two specimens) and group B (two specimens), respectively. MLST performed directly from cerebrospinal fluid revealed that the strains causing the two group C infections were of sequence type (ST) 11, while the two group B infections were characterized as ST-32 and ST-33 respectively. Multi-locus sequence typing of meningococci directly from clinical material offers the opportunity to improve further the surveillance of meningococcal disease and has now been introduced into the routine portfolio of tests employed at the national reference laboratory of the Czech Republic.

We investigated the prevalence and predictors of positive tuberculin skin test (TST) results among prospective Vietnamese migrants. We interviewed and medically screened 1395 Vietnamese people aged over 15 years who had applied to migrate to Australia. Approximately 44% of applicants had an induration of 10 mm or more, and 18·6% had an induration of 15 mm or more. A positive tuberculin skin test at 5 mm, 10 mm and 15 mm of induration cut-points was significantly associated with age (OR 1·01–1·02 per year) and duration of smoking (OR 1·03–1·12 per year). Smoking appears to be an important factor associated with increased susceptibility to mycobacterial infection. It is not yet clear whether the increased tuberculin reactivity associated with smoking reflects an increased risk of tuberculosis among these migrants.

The purpose of this study was to examine characteristic profiles of Streptococcus pyogenes clinical isolates isolated in Japan during 1994–9. Genotyping of the M protein (emm typing) revealed that emm types 12 and 28 were the most common among 316 isolates. Most of the emm12 isolates were isolated from mucosa, while emm58 and emm89 were from skin. Moreover, the emm3 isolates were dominant in invasive infections. The distribution of 6 superantigen genes showed that all isolates harboured the mf gene and many had the speG gene. Invasive isolates were shown to have the ssa gene at a higher rate (76%) than noninvasive (37%). The distribution of superantigens was significantly different between emm types, but not between isolation sites. These results suggest that the distribution of emm types is related to isolation site, whereas superantigen distribution is related to clinical features of S. pyogenes infections.

Two hundred and seven cases of listeria meningitis that occurred in The Netherlands over 20 years were reviewed to study associations between Listeria monocytogenes serotype, age, underlying disease, and outcome. The mean annual incidence per 100000 population was 0·12 in 1981–90, decreasing to 0·07 in 1991–5. Underlying disease was present in 50% of non-neonatal patients, most often haematological malignancy (15%) and the use of immunosuppressive therapy (14%). The meningitis-related case fatality rate was 16%; a significantly higher rate was associated with the presence of underlying disease (30%) or age [ges ]70 years (29%). Serotype 4b was most frequent (65%) and L. monocytogenes types 1/2a, 1/2b, or 1/2c (30% of cases) were significantly more often isolated from non-neonatal patients with underlying disease, suggesting a higher virulence of listerial serotype 4b.

We investigated which vaccination schedule gives best protection to the vaccinating population, in case there is a measles epidemic in an area with low vaccine coverage. We considered combinations of an early measles vaccination (none, at 6 months or at 9 months), a measles–mumps–rubella (MMR) vaccination around the first birthday (at either 11 or 14 months), and MMR vaccination at an older age (at either 4 or 9 years). The different estimates on measures of protection (percentage of susceptibles, number of reported cases in an epidemic year, percentage of lifetime spent susceptible) relied on a mathematical model of decline of maternal antibody levels with age, and the impact of that antibody level on seroconversion and immunity. Model parameters were estimated from a Dutch population-based serological survey on measles antibodies. Different measures of protection favoured different vaccination schedules, but dropping the age of second MMR vaccination prevents considerably more cases than an extra early measles vaccination or dropping the age of first MMR vaccination.