Various sampling techniques to determine the aerobic, E. coli and Entero-bacteriaceae counts and to detect the presence of salmonellas were compared. As a simple method for the detection of salmonellas a modified Surkeiwicz procedure using both drip and rinse water is advocated. To evaluate hygiene during processing, determination of the number of Enterobacteriaceae in pieces of skin from the ventral, lateral and breast region is preferred.

The immunizing activity of ribosome preparations derived from Brucella abortus strain 19 cells was examined in guinea-pigs and mice. After subcutaneous injections of Br. abortus ribosomes in Freund's incomplete adjuvant, both mice and guinea-pigs developed immunity to challenge by virulent Br. abortus 544 organisms which was at least as effective as the protection conferred by live strain 19 vaccine. Both mice and guinea-pigs also developed agglutinating and complement-fixing antibodies and delayed hypersensitivity to Br. abortus antigens. Conversely, ribosome preparations elicited delayed hypersensitivity reactions on intracutaneous injection into guinea-pigs chronically infected with Br. abortus or Br. melitensis.

On injection into rabbits, Br. abortus ribosomes incorporated in incomplete adjuvant induced high titres of agglutinins, complement fixing antibodies and precipitins for Br. abortus antigens. On immunochemical examination, the ribosome preparations were not grossly contaminated with antigens derived from the cell surface. They were chemically complex, however, and in addition to RNA contained numerous protein components identified by disk electrophoresis. The nature of the components responsible for conferring protection against Br. abortus was not determined.

Serum neutralizing antibodies to polioviruses were titrated in serum samples from 182 police cadets aged 16–18 years before and, in 168 of the cadets, 6 weeks after vaccination with a single dose of oral polio vaccine (OPV). Faecal excretion of poliovirus was also followed. Vaccination histories were obtained and confirmed whenever possible.

Pre-vaccination antibody could not be detected against type 1 in 9·3% cadets, against type 2 in 2·7% and against type 3 in 7·7%. Absence of antibody to at least one virus type was found in 14·3 % of the cadets.

In 93 cadets in whom vaccination histories could be confirmed 40 had received only inactivated polio vaccine (IPV) previously; of these 23% lacked antibody to at least one virus type, and they had less intestinal immunity to a challenge dose of OPV than those previously given OPV. Only two of the cadets known to have had OPV were non-immune – both had received a single dose following full courses of IPV. However, cadets who had received OPV had their last dose of vaccine more recently (average 4.6 years) than those who had received only IPV (all 12 years or more).

The serum antibody response to a single booster dose of OPV, and the faecal excretion of each type of virus after vaccination, showed an inverse relation to the corresponding pre-vaccination antibody concentration. A single dose of OPV did not reliably boost the immunity of those who possessed adequate immunity, and a failure to respond was also observed in a proportion of the cadets with no detectable antibody, mostly in the case of type 3 antibody and particularly if antibody to types 1 or 2 virus was also absent. No evidence was obtained that intestinal immunity could be expected in the absence of detectable circulating antibody.

The reasons for the absence of a serological response to OPV in some subjects are discussed and consideration is given to the practical significance of the findings. It is suggested that reinforcement of polio immunity at school-leaving is important, particularly at the present time when many of those aged 16–18 years will have been vaccinated only with IPV. A single does of OPV is not ideal for this purpose, not only because a small proportion of persons are liable to be left unprotected, but also because failure to produce a reliable boost in persons with adequate immunity at the time of vaccination gives rise to the possibility that they may become susceptible later in adult life.

After five months of total isolation a wintering party of seventeen British Antarctic Survey (BAS) personnel was inoculated under double blind conditions with placebo, or rhinovirus type 2 which had been propagated in tissue culture. The clinical and virological responses of these subjects were compared with those of volunteers in England who received a similar dose of the same strain. The virus used was apparently partly attenuated for man; at the dosage used its effects in England were similar to a smaller dose of an unattenuated strain, but in the Antarctic it caused relatively severe infections. Both the symptoms and the laboratory evidence of virus infection appeared to be more pronounced in the BAS subjects than in the volunteers in England who received the same challenge. In the former group the infection readily spread to those who were originally given placebo. In the BAS subjects serum antibody titres were well maintained during the isolation period but a significant fall in nasal immunoglobulin concentration was recorded during the 5 months of isolation after the virus challenge. Possible mechanisms for the increased sensitivity to rhinovirus of subjects who have been totally isolated in a small closed community are discussed.

The pathogenicity of two bovine field strains of virus for indigenous goats was examined in the laboratory. The goats failed to develop clinical disease or become virus carriers although the majority showed a definite immune response. A field survey in a foot-and-mouth disease enzootic area showed that the indigenous sheep and goat populations were frequently exposed to infection as evidenced by a high proportion of sero-positive animals but the incidence of virus carriers was very low in goats and no carriers were detected in sheep.

The antibacterial effectiveness of hand antiseptics commonly used in wards was studied by laboratory and in-use tests and their acceptability assessed by means of a questionnaire passed to hospital staff. To determine the immediate and long-term antibacterial effects of the preparations the in-use tests were performed by groups of students. The greatest immediate reduction in bacterial counts on hands was obtained by products containing chlorhexidine. The longterm antibacterial effect was recorded with emulsions containing 3% hexachlorophane, 2% Irgasan CF3R or 4% chlorhexidine when used constantly on several consecutive days. Considerable discrepancies were recorded in the antibacterial effectiveness of some preparations when comparing laboratory and in-use test results. Therefore it is suggested that antiseptics should be tested by in-use tests which more closely resemble practical conditions before their use, or further trial, in hospital.

Type 1 strains of Bordetella pertussis can infect mouse brain and have been recovered as type 1 organisms after death. When introduced into the naso-pharynx of the marmoset, they immediately acquired agglutinogen 2 or 3, and the resulting type 1,2 or 1,3 infection persisted for many weeks.

As in the child, agglutinogens 2 and/or 3 appear to be essential for infection of the marmoset, whereas they are quite unnecessary in mouse brain. A vaccine (extract or whole cell) containing agglutinogen 1 may be sufficient to pass the mouse protection test but it may fail to immunize children. The mouse test is inadequate even for the screening of such extracts.

A review of 194 cases of enteric infection in Scotland during 1967–74, is reported. Diagnosis was confirmed by the laboratory isolation of Salmonella typhi (64 cases), S. paratyphi A (3) and S. paratyphi B (127), from blood or faeces; 174 persons were clinically ill and 20 were symptomless infections.

Most patients (81·0%) were under 35 years of age, with the highest incidence occurring among young adults of 15–24 years, many of whom had been travelling overseas. One-third of all cases (65) were imported infections; the remaining 129 patients had not been outside the United Kingdom. A significantly greater proportion of typhoid infection (54·7%) was contracted overseas, in contrast to paratyphoid B with only one-fifth of cases (21·2%) being imported; all three cases of paratyphoid A were imported. The geographic distribution of origin of imported infections is discussed, along with the frequency of organisms belonging to different phage types.

Two deaths occurred, one of which was the result of complications of paratyphoid fever. All other patients responded well to treatment, although two persons continued to excrete and became chronic carriers.

A few examples are given of episodes of particular epidemiological interest.

It is expected that in future years there will be a continued increase in the proportion of imported infections as more persons travel overseas, concurrent with the continuing decline in the number of chronic carriers resident in the British Isles.

The serotypes of 13,139 strains of Escherichia coli isolated from humans were compared with the serotypes of 1076 strains isolated from animals. 689 of these strains were isolated from fresh cow-pats on 22 sites in England and Wales. 708 different O/H combinations were found. Of these, 520 were found in human strains only, 130 from animal strains only and 58 O/H serotypes from humans and animals. Approximately half of the animal strains could not be typed with the full set of sera used.

A schedule for the routine serotyping of strains of Bordetella pertussis based on agglutinin production in mice to the K-antigens has been worked out. Mice have been found as satisfactory as rabbits but far more economical for the production of the very small volumes of serum which are required. Agglutinin production, used in conjunction with direct agglutination, provides definitive information about serotype.

Sixty three samples of dairy creamery effluent were examined for the presence of mycobacteria. Thirty two strains were isolated from 27 samples. These were classified as follows: M. fortuitum (13), M. peregrinum (6), M. gordonae (5), M. marianum (scrofulaceum) (4), unidentified (4).

Ten strains, representative of the groups isolated, were tested for their effect on experimental animals. None were pathogenic for guinea pigs or mice but a number produced a minimal amount of skin sensitization in guinea pigs to avian and mammalian tuberculin.

Classification, by agglutinogens, of 634 isolates of Bordetella pertussis collected from 1941 to 1968 in Great Britain demonstrated that a change from a predominantly 1,2,0,4 serotype (75% of those examined during 1941–4) to a predominantly 1,0,3,0 serotype (73% of those examined during 1966–8) occurred sometime after 1953. Furthermore, evidence from the examination of isolates collected between 1941 and 1953 suggests that the change may have been gradual. Isolates of serotype 1,2,3,4 made up 20–30% of the total of our cross-country selection for the periods 1941–4, 1946–9, 1950–3 and 1966–8, but over shorter periods in individual areas the percentage varied from negligible to as high as half of those isolated. Results from other countries show a similar drift towards a 1,0,3 serotype but more often from a 1,2,3 than from 1,2,0 serotype.

The value, in epidemiological studies, of extended information obtained by monospecific typing sera to all six, rather than only two or three agglutinogens, and confirmation of the results by agglutinin production is demonstrated: for instance not all 1,0,3 isolates were identical.

Contamination of a laboratory environment with pathogenic or non-pathogenic micro-organisms may be relevant to safety of technicians and quality of technical performance. Two widely separated incidents in 1968 and 1974 initiated a study of aspects of the laboratory environment. Water-baths, water of syneresis and portions of salmonella cultures spurting out of the sterilizing flame were examined. The water of water-baths was shown to be contaminated from the fluid cultures incubated in them. This raised questions of potential cross-contamination and reporting of false positives. Water of syneresis was sometimes contaminated with salmonellas. A few quantitative counts were made. The range of counts varied between 16 salmonellas per ml. and 13,000,000 salmonellas per ml. Five hundred portions of salmonella cultures and 571 portions of Shigella sonnei cultures which had spurted from the sterilizing flame were examined. All these samples failed to grow salmonellas or shigellas.

Precautions necessary to avoid environmental contamination are briefly discussed.

An outbreak of shigellosis due to Shigella sonnei, is reported in laboratory maintained marmosets (Callithrix jacchus) and tamarins (Saguinus nigricollis). The clinical signs and pathological lesions are described and the microbiological findings discussed. Control of the disease was based upon hygiene and antibiotic therapy and the consequences of this approach are described in detail.

A thymine-requiring (thy−), trimethoprim-resistant (tmpr) mutant isolated from the faeces of chickens experimentally infected with Salmonella typhimurium and treated with a mixture of trimethoprim and sulphadiazine was less virulent for chickens than the parent strain and a thy+tmps revertant prepared in vitro from the mutant. The difference in chicken-virulence was more noticeable when the strains were administered orally than when they were administered subcutaneously. All tmpr mutants prepared in vitro from four other salmonella strains were also thy−; those tested were less virulent for chickens and mice than their parent strains. After oral infection, thy− salmonella organisms were found much less commonly in the alimentary tract of chickens then were thy+ organisms. This was especially so in the caeca, the principal site of colonization of both the thy+ and thy− organisms. Relatively high concentrations of thymine or related compounds were found in the contents of all regions of the alimentary tract of chickens except the caeca; the caeca usually contained low or undetectable concentrations.

The thy− salmonella strains would not grow on one brand of brilliant green agar because of its deficiency in thymine; their colonial appearance on other kinds of media used for isolating salmonellae from clinical material was often ‘un-salmonella-like’.

Strains of Bordetella pertussis in which all the organisms contain agglutinogens 1 and 3 or 1, 2 and 4 are easy to identify as serotypes 1,0,3,0 and 1,2,0,4 respectively; and similarly, stable strains of serotype 1,0,3,4 are occasionally found. During repeated subcultures, passage in vivo, and lyophilization and preservation for many years, these serotypes do not change. Mixing 1,0,3,0 and 1,2,0,4 serotypes and culturing them together in vivo and in vitro produces cultures from which organisms of the same two serotypes can be isolated.

In contrast, strains which type as 1,2,3,4 are often a heterogeneous group. We have attempted to classify these as 'stable', 'variable' and 'mixed' cultures. Some strains comprise organisms all of which contain the four agglutinogens and are as easy to type as the strains described above. These we have called 'stable' 1,2,3,4 strains. Other 1,2,3,4 strains are made up of colonies possessing all four agglutinogens, as shown by agglutinin production, but in amounts varying from day to day so that direct typing is inconsistent. These we have called 'variable' 1,2,3,4 strains. The last category, 'mixed', is made up of organisms most of which give rise to stable 1,2,3,4 cultures; a few of the component organisms, however, have one or two of the four agglutinogens missing. The importance of the 'variable' cultures is emphasized for work on apparent change of serotype, e.g. during infection.

The indirect immunofluorescent technique has been used to detect and titrate the specific immunoglobulins in serum specimens from 154 infants with confirmed or suspected congenital rubella. IgM antibody was stained more efficiently in sucrose density gradient fractions than in whole serum and was detected in this way in 27 out of 40 patients with confirmed congenital rubella at ages ranging from birth to 2 years. It was present in 48 out of 50 serum specimens during the first 6 months of life and in 11 out of 38 specimens obtained at ages between 6½ months and 2 years. IgM antibody was therefore estimated to persist for about 6 months in the majority of cases and up to 2 years in a few individuals. IgM antibody was also detected by this method in 11 out of 114 infants with suspected but unconfirmed congenital rubella at ages up to 5 months. The total concentrations of IgM were above the normal range in nearly all sera taken from confirmed cases during the first 3 months of life and in half the specimens obtained between the ages of 3 and 6 months.

IgG antibody was detected by fluorescent staining of whole serum in all patients with congenital rubella. Geometric mean titres increased during the first 3 months of life and then declined slowly. IgA antibody was not detected, except in two patients in whom traces were present at the age of 6 months, and the total concentrations of IgA were usually within normal limits.

Fluorescent staining of fractions showed that the sedimentation characteristics of rubella IgG and IgM antibodies were the same in infants as in adults. The peak IgM fractions never contained IgG antibody, and the presence of specific IgM in these fractions could usually have been safely inferred from their HAI titres. Fluorescent staining, however, was more sensitive and frequently detected IgM antibody in fractions which had no definite HAI activity.

Fluorescent staining of whole serum for IgM antibody was less distinct, and often unsuccessful, even in specimens in which specific IgM was detected in the fractions. The addition of IgG- to IgM-containing fractions caused depression of IgM staining and suggested that failure to detect IgM antibody in whole serum was partly due to competitive inhibition by specific IgG.

Defatting milk for the Brucella abortus whey agglutination test with chloroform caused some reduction in the titre in i.u./ml. in 42 of 57 samples and this reduction exceeded 50% in seven. Though this procedure will probably rarely affect the final interpretation of the test defatting the milk by centrifugation is preferred.

Brucella-specific antibodies of different immunoglobulin classes were quantitatively evaluated with respect to their efficiency in serological tests for bovine brucellosis.

IgM reacted more efficiently than IgG1 and IgG2 in both the Rose Bengal plate test and serum agglutination test. The complement fixation test was found to be slightly more sensitive to IgM than to IgG1 and did not react to IgG2. IgM was, however, partly inactivated when heated at 60°C. in the presence of serum.

Exposure of weaned hamsters to an environment contaminated with LCM virus shed by tolerantly infected mice led to short subclinical infections. If infection occurred in early pregnancy, the young appeared normal at birth but their tissues were highly infective. For two to three months their bites and urine were also highly infective. A viraemia did not persist long enough for successive vertical transmissions of the infection to be likely. However, the viruria persisted in most prenatally infected hamsters for at least eight months and under simulated field conditions was a potent virus source for contact infections, leading to further generations of prenatally infected young. In the absence of the natural reservoir host, such long-term carriers could have been a major factor in causing the build-up of infection in colonies of hamsters which, when purchased as household pets, led to a recent spate of human clinical infections in Germany and the U.S.A.