An enzyme immunoassay, with phenol–water-extracted lipopolysaccharide (LPS) from Brucella abortus as antigen, was used to detect the class-specific antibody response in sera from 173 patients with B. abortus, B. melitensis or B. suis infection. Sera from 30 patients with salmonellosis, yersiniosis or tularaemia and from 25 healthy individuals served as controls. The B. abortus LPS antigen permitted a safe diagnosis of acute and chronic brucellosis with high IgM and rising IgG titres in sera collected in the acute stage of the disease, and with elevated IgG titres only in the chronic stage. The B. abortus LPS antigen also permitted a specific diagnosis with the exception of the high titres estimated in sera from patients with Yersinia enterocolitica 09 infection. The problem with that well-known reciprocal cross-reactivity was overcome by using two additional antigens: Y. enterocolitica 09 native and periodate oxidized and borohydride reduced LPS preparations. In sera from patients with brucellosis high titres were estimated against all three antigens, whereas in sera from patients with yersiniosis caused by serotype 09 high titres were measurable only with the B. abortus and the Y. enterocolitica native LPS antigens. These data suggest that the B. abortus and Y. enterocolitica 09 LPS share one antigenic determinant resistant to periodate oxidation and borohydride reduction, and that in addition the Y. enterocolitica 09 LPS has a determinant which is sensitive to periodate oxidation and borohydride reduction.

The influence of multiple plating of fluid cultures on salmonella isolation from animal feeding stuffs was examined. Four platings were made from broth culture after 24 h at 37 °C and four platings from selenite enrichment from 24 h at 43 °C. Selenite enrichment followed broth culture which was used as a pre-enrichment stage. Brilliant green MacConkey agar plates were employed for broth subculture and brilliant green MacConkey and desoxycholate citrate agars for selenite subculture. The eight brilliant green plates subcultured from broth and selenite were examined for salmonellas after incubation for 24 h at 37 °C. The four desoxycholate citrate agars after 24 h at 37 °C were used for motility enrichment. The food sample size was a single 100 g instead of 4 x 25 g cultured in an earlier study. This pooling of samples aimed at technical economy. Quadruple plating played an important part in salmonella isolation from 100 g specimens. The combination of multiple plating with motility enrichment was the most successful technique used.

An IgM-antibody capture radioimmunoassay (MACRIA) was developed for the detection of IgM antibody specific for the human parvovirus-like agent B19. Diagnosis of infection with this agent by either antigen detection or antibody seroconversion had been made by counter-current immunoelectrophoresis (CIE) in 18 cases of aplastic crisis occurring in children with homozygous sickle-cell disease. The MACRIA described here gave positive results in 17 of these 18 cases; in the remaining case only an acute specimen taken from the patient during viraemia and late convalescent specimens taken 184 and 247 days after onset of illness were available.

The test was used to investigate 20 further cases of aplastic crisis in which neither viral antigen nor antibody seroconversion could be detected by CIE. Detection of virus-specific IgM permitted diagnosis of infection with this parvovirus-like agent in 17 of these cases. In the remaining three cases only single serum specimens taken late in convalescence, 82 days or more after the onset of symptoms, were available.

In addition to these 34 cases of aplastic crisis in which primary infection with this agent was diagnosed by MACRIA, seven cases of apparent ‘silent’ infection detected by CIE were investigated. The test permitted the discrimination between primary infection and re-exposure to the virus in six of these patients.

The use of this assay has added a considerable weight of evidence implicating primary infection with this parvovirus-like agent as an important cause of aplastic crisis in children with sickle-cell disease. Furthermore, MACRIA permits diagnosis of infection when only single serum specimens taken up to ten weeks after infection are available. Thus the use of this test will significantly facilitate the investigation of other clinical syndromes of presumptive infectious aetiology.

Measles virus-specific antibodies in sera from patients with HBsAg-negative chronic active hepatitis and raised antibody titres against measles virus, have been examined by crossed immunoelectrophoresis. The immunoprecipitates were further analysed by SDS-polyacrylamide gel electrophoresis. Five measles virus-specific precipitation lines were demonstrated using measles virus-infected cells solubilized with Triton X-100. The three major precipitation lines were analysed by SDS-PAGE and contained the virus polypeptides: nucleoprotein, NP (MW ∼ 60000); haemoagglutinin, H (MW ∼ 80000) and fusion protein F1 (MW ∼ 40000). Considerably higher amounts of antibodies against these three virus polypeptides were demonstrated in the patient sera than in sera from healthy controls. By SDS-PAGE analysis of radiolabelled immune complexes adsorbed to Sepharose-protein A, antibodies against five measles virus polypeptides: NP, H, F1, P protein (MW ∼ 70000) and matrix protein, M (MW ∼ 37000) were demonstrated in the patient sera.

In this study 197 strains of Escherichia coli isolated from cowpats and the faeces of healthy humans were examined for the presence of enterotoxins. Six strains representing five different serotypes were found to be enterotoxigenic. Four of these strains produced the heat labile enterotoxin and two, the Vero cell cytotoxin.

It is now established that milk can be a vehicle for the spread of enteritis due to Campylobacter jejuni. By determing the lethal effect of heat on six isolates it has been shown that C. jejuni is unlikely to survive pasteurization.

One hundred and eighty-two raw, 112 pre-cooked and 750 cooked hamburgers composed mainly of beef or beef and pork were subjected to microbiological examination.

Raw hamburgers gave total bacterial counts from 105 to 108 per g, counts of Enterobacteriaceae from 104 to 108 per g, of Escherichia coli from 103 to 105, of group D streptococci from 102 to 104, of Staphylococcus aureus from 3 to 102 and of Clostridium perfringens less than 10 bacteria per g. Of the samples, 32 % contained salmonellas; the highest most probable number was 102 per g but most estimates were below 1 per g. Corresponding figures for the pre-cooked samples were 2–3 log cycles lower, and only one sample contained salmonella. Yersinia enterocolitica was not isolated from any raw or pre-cooked sample.

Three hundred and ninety-five of the cooked hamburgers were prepared by grilling raw hamburgers for between 2 and 5·5 min. These gave total bacterial counts from 105 to 107 per g, and counts of Enterobacteriaceae from 102 to 105 per g. Of the samples, 9·4 % contained salmonellas, always in numbers below 1 per g. The remaining 355 cooked hamburgers were prepared from samples pre-cooked for 10 min at 80 °C. Some were grilled and some fat fried. The total bacterial counts were from 103 to 105 per g, and counts of Enterobacteriaceae below 102 perg. Salmonellae, again in small numbers only, were recovered from 3·5 % of samples.

When hamburgers were artificially contaminated with Salmonella typhimurium it took 5·5 min on a commercial grill, 2·25 min frying in a frying pan and 1·75 min on a household grill to reliably reduce the salmonella count one hundredfold. This means that at many vending places hamburgers are often cooked for too short a time.

D-values were determined for S. typhimurium in hamburger meat at 50, 55, 60, 65 and 70 °C, these values were 7·1, 5·1, 1·2, 0·9 and 0·6 min respectively. It can be concluded that the heating action in the centre of the hamburgers will take place more slowly than in the hamburger as a whole, and that the time between cooking and consumption is very important in reducing the microbial load to acceptable levels.

Pre-cooking (10 min at 80 °C in a water bath) gives a reduction in the numbers of salmonella of about 4 × 103, after which cooking gives a further reduction as mentioned above.

The single radial haemolysis (SRH) technique detected anti-B/HK/8/73 HA antibody rises in 59–85 % of paired sera from persons immunized with different influenza vaccines. In contrast, analysis of the same sera by the haemagglutination inhibition (HI) test indicated significant antibody rises in only 27–54% of paired sera. High levels of antibody were detected to influenza B/HK/8/73 and B/Singapore/222/79 viruses in post-immunization sera analysed by SRH, whereas the HI test indicated comparatively low geometric mean antibody titres. Most adults responded to immunization with influenza B virus by producing cross-reactive (CR) antibody which reacted with different influenza B viruses including the early isolate B/Lee/40.

The contamination of linen and air in a bed centre, supply station and ward were compared, as well as the contamination of gowns used by the staff working in the ‘clean’ and the ‘dirty’ rooms of the bed-centre. The contamination of linen and air was low and there was no significant difference between the tested areas. The contamination on gowns used by the staff working in the ‘dirty’ room was significantly higher than that on gowns used by the staff working in the ‘clean’ room. This stresses the importance of dividing a bed centre into ‘clean’ and ‘dirty’ rooms. So organized, a bed centre does not seem to be a hygienic hazard.

During the past year five patients from countries in the Middle East admitted to a burns unit were found to harbour a strain of Staphylococcus aureus with unusual multi-resistance to antibiotics. The admission of the first patient was followed by an outbreak of infection with this strain involving ten patients in the unit. In addition five staff members were found to be nasal carriers of the strain. As a result of this incident, the following four patients admitted to the unit were isolated on admission and the spread of their strains was thus prevented. It is recommended that patients on admission to burns units, or similar departments with patients very susceptible to infection, are isolated until their bacterial floras have been examined.

Mice were immunized by three intraperitoneal and one intravenous injection of rubella haemagglutinin. Splenocytes from these mice were fused with the cells of a syngeneic myeloma cell line, and following culture for various periods of time, single-cell clones were derived by the technique of limiting dilution.

A total of 139 clones were derived from 13 parent hybrid cultures. To date, four of these cloned cultures have been propagated as ascitic tumours in mice. The preparation of IgG from ascitic fluid and labelling of this antibody with 125I is described. Results indicate that the use of labelled monoclonal antibodies as indicator reagents in solid-phase IgM antibody capture assays for the detection of rubella-specific IgM results in enhanced performance of these tests.

An outbreak of suspected infectious hepatitis in a primary school was investigated using sensitive diagnostic methods for hepatitis A. A total of 116 sera from children were tested for the presence of both IgM and IgG antibodies to hepatitis A (HAV). The results were compared to those obtained for samples from a control school. IgG antibodies were present in 45 % and 10% of the children in the outbreak and control schools respectively.

A total of 2973 Escherichia coli, isolated from six different groups of animals, were examined for their ability to ferment adonitol, dulcitol, raffinose, rhamnose and sorbose in solid media. Twenty-nine fermentation patterns were recorded although 2443 (82%) of the E. coli belonged to seven of the 32 possible biotypes. Ninety-six O-serotypes were identified within the 2973 E. coli. The number of O-serotypes represented in the 15 most common biotypes ranged from three to 15. Sero types O8 and O9 were found most commonly in the different groups of animals and several biotypes amongst these two O-serotypes were identified in two or more groups of the animals. The ability of the E. coli to metabolize aesculin, ornithine, salicin and sucrose was also assessed. These tests proved less reproducible and were not included in the primary biotyping scheme although their use allowed the enumeration of additional biotypes. The application of biotyping to the study of the ecology of drug-resistant strains of E. coli in five situations is briefly presented.

Salmonella typhimurium phage type DT 193 survived in small numbers, in stored static slurry derived from veal calves, for the 7-week period of observation. The viable coliform count fell by 1½ logs during the first 2 weeks of storage, thereafter there were only relatively small fluctuations in the coliform population. In all 735 of 752 Escherichia coli isolates examined from eight samples of slurry were resistant to 3–6 antibacterial drugs. There was no dramatic change in the overall level of drug resistance amongst the E. coli with time. Chloramphenicol resistance was recorded in 400 (55%) of the E. coli. It was always associated with multiple resistance, with 96% of the strains being resistant to 5 or 6 drugs, although the proportion of isolates of each of the ten most prevalent O-serotypes resistant to chloramphenicol was variable and ranged between none and 97·5%. The use of biotyping together with O-serotyping indicated that the E. coli population was extremely complex, although certain components of the population remained relatively stable within the dominant flora with time since several of the more common O-serotype/biotype combinations were isolated from more than half of the eight slurry samples examined.

Antibody to cytomegalovirus (CMV) was sought in sera from Malta using immunofluorescence. Seven per cent of the infants, 36% of the school children, increasing to 100% of the adults aged over 40 years were found to have antibody. Most infection occurred in pre-school children and adults over 25 years of age. This pattern of antibody acquisition appears different from that described for other countries.

The virulence of faecal and urinary Escherichia coli strains was studied in relation to serotype, haemolysin production and haemagglutination pattern. By means of an experimental mouse model E. coli strains can be divided into avirulent (I), mouse nephropathogenic (II), and generally virulent (III) strains. Virulent group II and group III strains were more often haemolytic and haemagglutinating than avirulent group I strains. Presence of K antigen could not be associated with virulence. Discriminant analysis for qualitative variables revealed that no combination of the investigated properties contributed more to a strain's virulence level than did one single property. It is concluded that other virulence factors, apart from haemolysin production in group II strains and haemagglutinins in group III strains, must be involved in the determination of a strain's virulence level.

All O2, O6 and O18ac strains tested were virulent, and by far the most O75 strains were avirulent, whereas other O groups were more variable with regard to virulence. Pyelonephritis strains were more often mannose-resistant haemagglutinating than faecal and other urinary isolates, indicating that mannoseresistant adhesins may be important in the pathogenesis of pyelonephritis.