The sequential acquisition of bacterial flora by premature neonates was studied during a 10 month period. Mean gestational age of the babies was 29·01 weeks and the mean birth weight was 1·728 kg. Escherichia coli and group B streptococci (GBS) colonized the umbilicus of 7 and 6 babies respectively, out of 23 studied, on the first day of life. E. coli and staphylococci were the predominant flora on the 6th day and they colonized 12 and 13 respectively. The oral flora was predominantly Gram-positive cocci, mainly Streptoccocus salivarius which was isolated from 17 out of 22 babies on the 6th day, viridians streptococci were isolated from 14 babies, Staphylococcus albus from 16 babies and group D streptococci from 11 babies. Candida spp. also colonized the oral cavities of 17 out of 22 babies on the 6th day. At the end of the first week of life, the faecal flora was predominantly anaerobic represented by Bifidobacteriurn spp., Bacterioides spp. and Clostridium spp. The commonest facultative faecal flora were E. coli, which was isolated from all the babies, and Strept. faecalis isolated from 20 babies. Early gut colonization by GBS, Bacteroides spp. and Clostridium spp. was noticed in more babies delivered vaginally than by caesarean section where colonization by these bacteria was relatively delayed. The use of prophylactic penicillin plus gentamicin in the special neonatal unit probably prevented systemic spread of any of the potential opportunistic pathogens during the study.

A collection of 50 bacteriocins was assembled and used to type 802 isolates of Clostridium perfringens from food poisoning outbreaks and a variety of other sources. It was found that strains of the same serotype within an outbreak showed similar patterns of susceptibility to bacteriocins, and the use of a ‘one difference’ rule is proposed for interpretation of the typing patterns of epidemiologically related strains. Isolates of different serotype or of the same serotype isolated from different sources produced many variations in bacteriocin susceptibility patterns.

Two computer programs were developed to assist in the interpretation of bacteriocin typing patterns. Their use showed that related and unrelated strains formed different clusters and enabled a range of the 20 most discriminatory bacteriocins to be selected.

Isolates of C. perfringens from a wide range of sources were screened for their ability to produce bacteriocins. A much greater proportion of the strains from food poisoning outbreaks was bacteriocinogenic than were isolates from human and animal infections, various foods and the environment. The relevance of these findings to the occurrence of C. perfringens food poisoning is discussed.

In an inter-laboratory survey, 148 samples of cooked prawns and shrimps were obtained at the point of sale to the consumer. Salmonellae and Vibrio parahaemolyticus were not detected. Yersinia enterocolitica was isolated from three samples. Results for total viable count and presence of Escherichia coli and Staphylococcus aureus complied well with available guidelines for imported cooked prawns, suggesting that the risk of food poisoning from retail samples of these foods in the South of England is minimal.

We describe a community outbreak of campylobacter enteritis associated with the consumption of untreated milk, apparently contaminated by two cows with campylobacter mastitis. The outbreak occurred in two phases. Strains of Campylobacter jejuni of the Penner serogroup complex 4, 13, 16, 50 and Preston biotype code 6100 were isolated from patients in both episodes and from the faeces of the cattle, milk filters, bulk milk and retail milk. Milk samples from two of 40 milking cows were found to contain C. jejuni, and the wheys from these two cows had high titres of antibody to C. jejuni detected by ELISA techniques.

Less than one per cent of serum samples taken from 257 fish farmers in 1981 had agglutinating antibodies to strains of Leptospira interrogans of serogroup Icterohaemorrhagiae at a titre of 30 or greater. Compared with the results from other serological surveys, this agglutinating antibody prevalence suggests that fish farming does not have a high occupational risk for leptospirosis. Between 1961 and 1981 the incidence in fish farmers was about 33 per 100000 person years at risk. During the same period the incidence in the general adult male population was 0·137 per 100000 person years at risk, so that fish farming had a moderately increased risk of Icterohaemorrhagiae serogroup infection (relative risk = 243). No one particular risk factor within fish farming could be reliably identified and therefore recommendations to reduce the risk can only be general.

Coumatetralyl was evaluated in the laboratory against Bandicota bengalensis and Rattus rattus. Feeding tests with 0·0375 % coumatetralyl baits produced 100 % mortality after a single day feeding period in B. bengalensis and after a 10–day period in R. rattus. The anticoagulant bait is less palatable in comparison to plain bait. In the case of R. rattus, LFP50 and LFP98 and their 95 % confidence limits were 3·89 (2·62–5·77) days and 11·22 (6·1–20·65) days respectively. Median period of survival and its 95% confidence limits of B. bengalensis and R. rattus were 4·7 (3·85–5·7) days and 11·2 (9·33–13·44) days respectively.

A μ-capture ELISA was developed for detecting Mycoplasma pneumoniae specific IgM, and compared with an indirect immunofluorescent antibody (IFA) technique and an indirect ELISA. μ-capture ELISA and IFA compared well and were found to be the most sensitive assays. The IFA test can be completed in 2 h whilst the results of the μ-capture ELISA can be available in 24 h. Both tests are amenable to routine diagnostic use and have similar sensitivity. Indirect ELISA was found to be less sensitive and less specific, giving high assay values with several sera having undetectable M. pneumoniae CF antibody or CF antibody in low titre. Serum samples obtained from 11 patients at various times after M. pneumoniae infection showed maximum antibody levels within the first month by all assays, with a gradual fall in amount of IgM with time when assayed by μ-capture ELISA, a more gradual decline by IFA and hardly any decline with indirect ELISA. It was concluded that the indirect ELISA is unsuitable for the investigation of possible M. pneumoniae infection because the sustained high assay values with serum samples taken many months after infection, make interpretation of the test results very difficult.

Three influenza A (H3N2) reassortant whole virus vaccine strains with differing antibody-inducing capacities in hamsters were investigated morphologically and antigenically. Although initial measurements of virion circumference, from electron micrographs of vaccine preparations, suggested a relationship of small virion size with low immunogenicity, subsequent immunization with, and morphological investigation of, vaccine virions separated on sucrose gradients, failed to obtain populations whose antibody-inducing capacity clearly correlated with constituent virion density, size, morphology or integrity.

However, antigenic investigation using single radial haemolysis (SRH) and monoclonal antibodies revealed significant differences in antigenic specificity between the strains. Furthermore, a series of H3N2 isolates, derived using standard reassortment procedures, also showed differences in antigenic specificity in their haemagglutination-inhibition (HI) reactions with monoclonal antibodies after five passages in allantois-on-shell cultures. Variation between these isolates and their A/Victoria parent virus could be detected using SRH and hamster sera raised against each isolate.

These results demonstrate variation between candidate influenza A virus vaccine strains, all possessing the same surface (H3N2) glycoproteins, expressed as a consequence of the reassortant system used for their production.

During the winter of 1982 concurrent outbreaks of influenza A and influenza B occurred. The epidemiology and clinical features of 151 cases referred during this time are described, and patients are discussed according to age and presenting clinical syndrome: croup was the commonest presentation in young children, a typical influenza syndrome predominated in young adults, while older patients were more likely to have lower respiratory tract infection. There was no significant difference between the clinical features of influenza A and influenza B. Unusual clinical features include rash, exudative tonsillitis and the need for myringotomy during the course of influenza.

The serum antibody responses and 50% protective levels (PL50) of antibody were determined, using the SRH test, at one and twelve months post-vaccination in a group of student volunteers immunized with one of three dosages of a trivalent surface-antigen influenza virus vaccine, or with placebo.

It was found that, for the H3, H1 and B haemagglutinin components present in the vaccine, a dose of 6 μg HA elicited high serum antibody responses at one month post-immunization. High mean antibody levels and a high incidence of volunteers with PL50 values of antibody against each of the HA components of the vaccine remained in the volunteer group twelve months later. The results are discussed in relation to the vaccine dosage used and the nature of the population immunized.

Five hundred and seventy-one camel sera collected after an epizootic of Rift Valley Fever were examined for antibody to the virus. Clinical disease had not been observed in cattle and sheep in the ecosystems shared with the camels. Positive sera with high titres of serum neutralizing antibody were found in 22% of camels at one of the seven sampling sites.

Ether-treated influenza B haemagglutination inhibition (HI) antigen was used in a study of serum collections from three different epidemic seasons.

For diagnostic purposes, ether treatment increased the efficacy of the HI test by about 50 % over the conventional HI technique, raising it to the same level of sensitivity as the complement fixation (CF) test. The treatment reduced the specificity of the HI test, but its reliability in the diagnosis of influenza B infections was only slightly diminished. With regard to evaluation of the immune status of a given population, an HI test using ether-treated antigen from the epidemic influenza B strain seems to give more relevant information about the antibody level associated with protection than a conventional HI test using untreated virus antigen.

Fourteen influenza A(H3N2) field isolates, mainly obtained during the first weeks of the 1982/83 influenza epidemic in The Netherlands, and nine influenza A(H3N2) reference strains were examined by means of haemagglutination inhibition (HI) tests with 23 potyclonal ferret sera. The resulting HI patterns were subjected to various methods of numerical taxonomy using, among others, taxonomic distance and correlation between strains for resemblance coefficients. Marked differences between distance and correlation coefficients were found in strains which differed in avidity only. The field isolates could be divided into four groups in respect of their taxonomic resemblance to the reference strains. The same grouping was found for five of the field isolates by testing these against 200 human sera.

Seven virus strains antigenically related to the California encephalitis (CE) virus group were isolated from Norwegian Aedes spp. mosquitoes collected in 1976. So far CE viruses have been isolated from five different Aedes spp. in Norway. Furthermore, two virus strains related to the Bunyamwera group were isolated from Anopheles claviger.

Antibodies to CE viruses were demonstrated in 22% of 1014 military recruits tested. Among 91 soldiers who were monitored by monthly blood samples during the mosquito season, seroconversions were detected in 11 individuals. Specific IgM antibodies were found in seven of them. Disease symptoms in connexion with the CE virus infections were not seen. The prevalence of CE antibodies in patients with CNS or respiratory infections was not higher than in control groups. Seroconversions were not seen in any of the groups.

Screening of sheep sera from six different areas in northern Norway indicated significantly different degrees of CE virus activity. Passerine birds may be important CE virus hosts, while small rodents seem unimportant. Specific IgM antibodies were detected in the sera of one of three hares and one of two squirrels.

Of the methods used, single radial haemolysis (SRH) and immuno-electroosmophoresis (IEOP) seemed to be well suited for serological screenings. However, an indirect immunofluorescence antibody test (IFAT) which was used may be an attractive alternative if high-quality anti-species conjugates are available. The haemagglutination-inhibition (HI) test used gave a high number of false positive results.

Surveillance of certain respiratory viral infections by applying immunofluorescence (IF) examinations to samples of nasopharyngeal secretions has been evaluated using a simplified procedure for the preparation of cell smears. Samples from 711 children living in different parts of Norway were examined during the winter 1982/83 and a positive diagnosis was made for 290 children (41%). Temporal epidemic peaks were observed for respiratory syncytial virus (RSV), parainfluenza virus type 3 and influenza virus. On the other hand, the monthly number of negative samples was almost constant throughout the period. Differences in timing of RSV outbreaks were observed between two regions in Norway. Compared to rapid IF diagnosis, RSV notifications obtained by serological examinations were delayed by several weeks. Rapid virus diagnosis by IF examinations with our simplified procedure for preparation of nasopharyngeal samples seems to be suitable for the epidemiological surveillance of respiratory viral infections, both for its simplicity of preparation of the samples and for its accuracy in defining the time of the actual virus infection. Nevertheless, the method is not without pitfalls; a close cooperation between those who take the specimens and the laboratory is essential, and the IF examinations should be performed by an experienced microscopist.

Small rodents were collected live in two different locations within a nephropathia epidemica (NE) endemic area, and tested for both antiviral serum antibodies and viral antigens in lung sections. In one location, only Apodemus sylvaticus (woodmice) were found in the traps, in the other, both A. sylvaticus and Clethrionomys glareolus (bank voles) were collected.

Among the woodmice from the former location the prevalence of NE virus markers was significantly lower than for either woodmice or bank voles from the other location, and no NE antigen-positive animals was found. The woodmice co-existing with bank voles had a lower prevalence of NE antigen and antibodies than the bank voles, and fewer woodmice had both antibodies and antigen. The results emphasize the important role of bank voles as a major NE virus reservoir and probable source of human infections.

Recent evidence of bluetongue (BT) virus infection of livestock in scattered localities in the neotropics prompted a serologic survey of cattle in Colombia and Costa Rica. In Costa Rica 48·1% of 1435 bovine animals had BT virus antibody in the agar gel precipitation test (AGPT). In Colombia 51·8% of 635 cattle were AGPT-positive for BT virus. Antibody prevalence ranged from over 50% in the lowlands to 0% in Costa Rica and 19% in Colombian cattle above 2000 m altitude. Neutralization tests indicated that Costa Rican cattle had been exposed to BT virus types 6, 12, 14 and 17.

Formalin-fixed samples from 221 animal brains received for rabies diagnosis in Nigeria were digested in 0·1% trypsin in phosphate buffered saline, pH 7·4, and smears stained for rabies antigen by direct immunofluorescence (IF). The results were compared with those obtained using fresh material from the same animals for Negri body staining, mouse inoculation (MI) and occasionally immunofluorescent staining.

From 191 specimens examined for Negri bodies and by mouse inoculation 51 and 64 respectively proved positive. The IF smear technique under investigation failed to detect 5 of these but showed up as positive 30 which had been recorded as Negri-negative and 19 that had gone undetected by MI too. In a direct comparison with IF staining of fresh tissue from 23 known rabies positive animals the similar staining of trypsin-digested formalized smears failed to give a positive result in 2 out of 23 cases.

Some problems were encountered with smears not sticking to slides. When gelatinized slides were used fluorescence was sometimes poorer.

Where transport and refrigeration are difficult and section-cutting equipment is lacking the technique of IF staining of smears prepared from formalized brain tissue after treatment with trypsin can be a useful adjunct to other diagnostic methods. It also makes for safer working where special facilities are absent.