A variant of variola virus is described which produces a late polypeptide of 25 kDa instead of one of 27 kDa and which has an additional endonuclease cleavage site for SalI in the viral DNA. These markers were shown to be genetically independent and to characterize 14 of the 48 variola strains which were examined. The variant strains were isolated from smallpox outbreaks originating in or from Pakistan between 1961 and 1974 and also from two cases at a Mission Hospital in Vellore, India in 1964. No variant strains were found among 9 other isolates from cases of variola major occurring in other parts of India or in Bangladesh, nor among 4 isolates from Indonesia, 15 from Africa or 6 isolates of variola minor.

Haemorrhagic fever with renal syndrome (HFRS) is a human disease characterized by flu-like symptoms, renal dysfunction, and in severe cases, haemorrhagic manifestations. The causative agents of HFRS are Hantaan (HTN), Seoul (SEO), Puumala (PUU) and Dobrava (DOB) hantaviruses. Hantavirus infections are of increasing importance in Europe. Outbreaks occur in Belgium with a 3- to 4-year interval with an increasing number of cases. We describe the largest outbreak so far in Belgium with 217 serologically and clinically confirmed cases in the period between October 1995 and December 1996. We demonstrated that the use of viral antigen derived from a local PUU-strain was able to detect significantly more sera positive for IgM in an immunofluorescence assay. Furthermore, although in some cases SEO, HTN and DOB antibody-reactivities were detected by ELISA, only PUU infections could be confirmed by neutralization test. The presence of an unknown hantavirus serotype circulating in Belgium should be considered.

It is generally accepted that the reservoir hosts of cowpox virus are wild rodents, although direct evidence for this is lacking for much of the virus's geographic range. Here, through a combination of serology and PCR, we demonstrate conclusively that the main hosts in Great Britain are bank voles, wood mice and short-tailed field voles. However, we also suggest that wood mice may not be able to maintain infection alone, explaining the absence of cowpox from Ireland where voles are generally not found. Infection in wild rodents varies seasonally, and this variation probably underlies the marked seasonal incidence of infection in accidental hosts such as humans and domestic cats.

A cross-sectional serosurvey for markers of prior Cryptosporidium infection was conducted among homosexual or bisexual males infected with human immunodeficiency virus (HIV); of 262 individuals approached, 236 (90%) agreed to participate. Serological response to two Cryptosporidium antigens was measured using a Western blot assay. The intensity or detection of serological responses to two Cryptosporidium antigens was not associated with CD4 cell counts or tap water consumption. A number of sexual practices were related to increased serological response for only the 27-kDa marker, including having had sex within the past 2 years, having anal sex and having had a larger number of sex partners during the past 2 years. Attending a spa or sauna was related to serological response to both the 27-kDa and 17-kDa markers. Based on these results, activities related to sexual activity appear to be a significant risk factors for prior Cryptosporidium infection.

An automated early warning system has been developed and used for detecting clusters of human infection with enteric pathogens. The method used requires no specific disease modelling, and has the potential for extension to other epidemiological applications. A compound smoothing technique is used to determine baseline ‘normal’ incidence of disease from past data, and a warning threshold for current data is produced by combining a statistically determined increment from the baseline with a fixed minimum threshold. A retrospective study of salmonella infections over 3 years has been conducted. Over this period, the automated system achieved >90% sensitivity, with a positive predictive value consistently >50%, demonstrating the effectiveness of the combination of statistical and heuristic methods for cluster detection. We suggest that quantitative measurements are of considerable utility in evaluating the performance of such systems.

The relationship between ocular toxoplasmosis and levels of toxoplasma specific antibodies was examined in 195 patients. Using clinical information collected by questionnaires, patients were divided into: 97 with ocular toxoplasmosis (group 1) and 98 with ocular lesions not due to toxoplasma (group 2). The geometric mean of dye test titres (±S.D. natural log titre) in group 1 was 53·2 (±0·95) compared with 24·6 (±1·11) in group 2 (P<0·001). Young females tended to have more active lesions compared with young males (P<0·05). There was an age-dependent difference in dye test titres between the groups (P<0·001). Group 1 showed a decline in titre with age compared with an increase in group 2. Ocular toxoplasmosis was diagnosed most frequently among 21–30 year olds. More group 1 patients had dye test titres [ges ]65 iu/ml than group 2 (P<0·05). Dye test titres [ges ]65 iu/ml support a diagnosis of ocular toxoplasmosis whereas lower titres suggest other causes for eye lesions.

This cross-sectional study of 400 sera from a randomly selected adult population in Northern Ireland, using a microimmunofluorescence assay, demonstrated high overall seropositivity (70%) for IgG Chlamydia pneumoniae antibodies in developed populations. Seropositivity was shown to be unrelated to gender, age or smoking but there was an inverse trend between infection and educational level achieved as a measure of socio-economic status. IgG levels were also higher during the winter months suggesting seasonal variation of Chlamydia pneumoniae infection. The high prevalence of evidence of exposure to Chlamydia pneumoniae as described in this study may have implications for prevention of cardiovascular disease if further evidence conclusively determines that infection with this organism is a risk factor for cardiovascular disease.

A study was conducted to characterize the risk of Plasmodium vivax infection in the Lacandon forest, southern Mexico. Blood samples and questionnaire data were collected in 1992. Malaria cases (n = 137) were identified by the presence of symptoms and a positive thick blood smear. The control group included individuals with negative antibody titres and no history of malaria (n = 4994). From 7628 individuals studied, 1006 had anti-P. vivax antibodies. Seroprevalence increased with age. Risk factors associated with infection included: place of birth outside the village of residence (odds ratio, OR 11·67; 95% CI 5·21–26·11); no use of medical services (OR 4·69, 95% CI 3·01–7·29), never using bed-nets (OR 3·98, 95% CI 1·23–12·86) and poor knowledge of malaria transmission, prevention and treatment (OR 2·30, 95% CI 1·30–4·07). Health education represents the best recommendation for controlling the disease in the area.

The newly described microorganism ‘Simkania Z’, related to the Chlamydiae, has been shown to be associated with bronchiolitis in infants and community acquired pneumonia in adults. The prevalence of infection in the general population is unknown. A simple ELISA assay for the detection of serum IgG antibodies to ‘Simkania Z’ was used to determine the prevalence of such antibodies in several population samples in southern Israel (the Negev). The groups tested included 94 medical and nursing students, 100 unselected blood donors, 106 adult members of a Negev kibbutz (communal agricultural settlement), and 45 adult Bedouin, residents of the Negev. IgG antibodies to ‘Simkania Z’ were found in 55–80% of these presumably healthy individuals, independently of antibodies to Chlamydia trachomatis and Chlamydia pneumoniae. The Bedouin had a seropositivity rate of 80%, while all other groups had rates of between 55 and 64%. These results indicate that ‘Simkania Z’ infection is probably common in southern Israel.

Anecdotal evidence suggests that the prevalence of both scabies and head lice is increasing and also that both conditions are becoming refractory to pesticide treatment. Using information obtained from the Office of National Statistics, Royal College of General Practitioners Weekly Returns Service, Department of Health, local surveys of school children from Bristol and drug sales of insecticides, we have confirmed that there has been a rise in the prevalence of both conditions. We have shown that scabies is significantly more prevalent in urbanized areas (P<0·00001), north of the country (P<0·000001), in children and women (P<0·000001) and commoner in the winter compared to the summer. Scabies was also shown to have a cyclical rise in incidence roughly every 20 years. Head lice were shown to be significantly more prevalent in children and mothers (P<0·000001) though both conditions were seen in all age groups. Head lice were also less common during the summer. Host behaviour patterns, asymptomatic carriage, drug resistance and tourism from countries or districts with a higher incidence may be important factors in the currently high prevalence of both scabies and head lice.

The world's first documented toxoplasmosis outbreak associated with a municipal water supply was recognized in 1995 in Victoria, British Columbia, Canada. It was hypothesized that domestic cat (Felis catus) or cougar (Felis concolor) faeces contaminated a surface water reservoir with Toxoplasma gondii oocysts. An extensive investigation of the Victoria watershed 1 year following the outbreak documented the presence of an endemic T. gondii cycle involving the animals inhabiting the area. Cats and cougars were observed throughout the watershed. Serological evidence of T. gondii infection was demonstrated among domestic cats living in the Victoria area. Cougars were found to shed T. gondii oocysts. Serological evidence of T. gondii infection in deer mice living in the riparian environments of the watershed suggested that T. gondii oocysts were being shed near the water edge. Contamination of Victoria's water supply with T. gondii oocysts potentially occurred during the study period and future waterborne toxoplasmosis outbreaks in this and other communities are possible.

Escherichia coli O157 (n = 376) from 41 cattle farms were subtyped using pulsed field gel electrophoresis of endonuclease cleaved chromosomal DNA. Cleavage with XbaI resulted in 81 subtypes. Fifty-one isolates from subtypes found in more than one herd, or in herds on multiple sample collection dates were compared using the endonuclease NotI, resulting in 23 additional subtypes. Up to 11 XbaI subtypes were found per farm with up to 7 subtypes/farm identified from a single date. Indistinguishable subtypes (both XbaI and NotI) were found to persist on 4 farms for 6–24 months. Five subtypes were found on more than one farm separated by up to 640 km. Dairy farms where cattle had moved onto the farm had a similar number of subtypes as farms with no movement of cattle, and feedlots had more subtypes than dairy farms. These data indicate that there is a mechanism for multiple herd exposure to specific subtypes, there are multiple sources of exposure for cattle on farms, and on-farm reservoirs other than cattle may exist.

Although information on amoebiasis among institutionalized psychiatric patients is available, reports on the relationship between behaviour and this infection are not abundant. From July 1995 to June 1996, stool and blood samples were collected from 565 patients in three psychiatric hospitals of North Taiwan. Stool samples were examined using the direct smear and formalin-ethyl acetate sedimentation techniques as well as ProSpecT® Entamoeba histolytica Microplate Assay kit. Blood samples were examined by the Amebiasis Serology Microwell ELISA kit. Among these patients, 14 (2·5%) harboured one or two species of intestinal parasites. There were 6 (1·1%) E. histolytica/E. dispar cyst passers: 5 positive in stool ELISA test and 2 with antibodies against E. histolytica. Among demographic factors, type of psychiatric disorder and disability, only a significant sexual difference in seropositivity of E. histolytica was observed. These findings indicate that the infected patients acquired the infections before they entered the hospitals.

Confirmed acute hepatitis B infections are reported to the Public Health Laboratory Service Communicable Disease Surveillance Centre by laboratories in England and Wales. These reports have been used to monitor trends in the incidence of hepatitis B virus (HBV) infection over time, and between exposure categories and age groups. Between 1985 and 1996 a total of 9252 cases of acute HBV infection were reported; the number of reports fell from 1761 in 1985 to 581 in 1996. Most infections were reported in adults aged 15–44 years [n=7365 (80%)], and infections were more commonly reported in males [n=6490 (70%)] than females [n=2658 (29%)]. The probable means of acquisiiton was known for just over half of all adult cases [4827/8956 (54%)]. Injecting drug use was the most common exposure [n=1901 (21%)], followed by sex between men and women [n=1140 (13%)] and sex between men [n=1025 (11%)]. The number of infections in injecting drug users fell in the late 1980s, but increased again from 1991 onwards. In children aged under 15 years, infections acquired by mother to baby transmission accounted for 35/170 (21%) of the total. Surveillance indicates that the incidence of acute hepatitis B infection fell in the late 1980s, probably reflecting changed behaviour in injecting drug users. An increase in the number of infections in injecting drug users since 1993 may indicate ongoing transmission that has not been contained by the introduction of needle exchange schemes or by selective vaccination.

The objective of this study was to determine the most efficient means of sampling faeces of finisher pigs for accurate and precise farm-level estimates of antimicrobial resistance among faecal Escherichia coli. Resistance to tetracycline and gentamicin of 8250 isolates of E. coli from 55 finisher pigs on one farm was measured with a hydrophobic grid membrane filter method. The between-pig, within-pen component of variance in resistance was large (97·5%), while between-pen, within-room and between-room components were small (2·5% and 0%, respectively). Using these resistance data, the abilities of two sampling strategies to estimate prevalence were modelled with a Monte Carlo ‘bootstrap’ procedure. Compositing faecal samples from several pigs before testing produced unbiased and precise estimates of prevalence and is simpler technically than individual animal testing.

Cost effectiveness analyses of alternative hepatitis B vaccination programmes in England and Wales require a robust estimate of the lifetime risk of carriage. To this end, we report the prevalence of infection in 3781 anonymized individuals aged 15–44 years whose sera were submitted in 1996 to 16 microbiology laboratories in England and Wales. One hundred and forty-six individuals (3·9%) were confirmed as anti HBc positive, including 14 chronic carriers (0·37%). The prevalence of infection and carriage was higher in samples collected in London and increased with age. No increased risk of infection was seen in sera from genito-urinary (GUM) clinics. Only 15 sera positive for hepatitis B were also positive for hepatitis C. Our results confirm the low prevalence of hepatitis B in England and Wales, are consistent with previous estimates of carriage and suggest that many infections were acquired while resident outside the UK. Future prevalence studies should determine the country of birth and other risk factors for each individual in order to confirm these findings.

A panel of 16S ribosomal RNA gene probes has been developed for the study of the epidemiology of heartwater; five of these detect different cowdria genotypes, one detects five distinct genotypes; one detects any Group III Ehrlichia species other than Cowdria and one detects any Group II Ehrlichia species. These probes have been used on PCR-amplified rickettsial 16S rRNA genes from over 200 Amblyomma hebraeum ticks. Control ticks were laboratory-reared and either uninfected or fed on sheep experimentally infected with different cowdria isolates, field ticks were collected from animals in heartwater-endemic areas. All tick-derived DNA samples were also examined by PCR amplification and probing for two other cowdria genes (map1 and pCS20) which have previously been used for heartwater epidemiology. This paper describes the first direct comparison of all currently available DNA probes for heartwater-associated organisms.

The objectives of this study were to investigate the coagulase gene polymorphism of Staphylococcus aureus isolates obtained from bovine mastitic milk and to determine the resistance of predominant and rare coagulase genotypes to bovine blood neutrophil bactericidal activities. A total of 453 isolates were collected from four countries: the Czech Republic, France, Korea and the United States. The isolates were subtyped into 40 types by restriction fragment length polymorphism (RFLP) of the coagulase gene. Twenty-three strains from predominant and rare genotypes were evaluated for their ability to resist neutrophil bactericidal activities. There were significant (P<0·01) differences in the average percent neutrophil killing of the predominant (16·7%) and rare (39·7%) genotypes when bacteria were opsonized with antiserum. The results indicate that the profiles of coagulase genotype differ among geographic locations, and only a few genotypes prevail in each location. In addition, the predominant genotypes were more resistant to neutrophil bactericidal activities than rare genotypes.

We studied the seroprevalence of hepatitis B virus (HBV), hepatitis C virus (HCV) and GB virus-C (GBV-C) infections in 348 Siberian natives who lived in the Kamchatka Peninsula of Russia. Of 348 samples studied, the seroprevalence of HBsAg and anti-HBs were 11·8% (41 of 348 samples) and 35·9% (125 of 348 samples), respectively. The prevalence of HCV infection was 1·4% (5 of 348 samples), and that of GBV-C RNA, using RT–PCR methods, was 7·5% (26 of 348 samples). In Siberia, the prevalences of HBV and GBV-C infections were about tenfold higher than those in Japan. The prevalence of HBsAg in subjects under 50 years of age was significantly higher than that in those over 50 years old (P<0·05). Because HBV infection is highly endemic in Siberia, we propose that the community-based mass immunization must be conducted as soon as possible in this area.

Salmonella enterica serovar Infantis is endemic in Finnish cattle. Feed contaminated with S. Infantis was distributed to cattle farms in May 1995. Following increased sampling, S. Infantis was detected on 242 farms in 1995. Molecular typing was used to differentiate the farms that were infected by the feed-related Infantis from those infected by other endemic strains. Twenty-three isolates from feed in 1995 and 413 from cattle (72 from 1992–4, 324 from 1995, 17 from 1996–7) were analysed. The feed-related Infantis was clonally related to the endemic infection by the ribotype, IS200-type and XbaI-profile. The feed isolates had a distinctive plasmid that appeared in pulsed-field gel electrophoresis as a 60 kb band when cleaved with XbaI or linearized by S1-nuclease. This plasmid appeared in cattle only since the outbreak and seemed stable on the follow-up farms. In addition to contact farms, the feedborne strain was found on 19% of the farms infected with S. Infantis in 1995 but not having bought suspected feedstuffs, possibly as secondary infections.