A three-part epidemiological investigation was made on flaviviruses:

1. As a preliminary to tests on cattle sera from the field, the antigenic cross-reactivity of Wesselsbron, Spondweni, Usutu, Banzi, West Nile and yellow fever flaviviruses was studied in antisera prepared in guinea pigs. As described earlier for flaviviruses, sera were found to be highly cross-reactive in haemagglutination-inhibition (HAI) tests, less cross-reactive in complement-fixation (CF) tests and were virtually monospecific in microneutralization (NT) tests in Vero cell cultures.

2. Infection with Wesselsbron (WSL) virus produced mild febrile illness and viraemia in 5 out of 6 newborn calves, 3 out of 4 pregnant heifers and 3 out of 4 ewes. One heifer produced a weak calf which died soon after birth with WSL antibodies in its serum, indicating that infection had occurred in utero. The 3 other heifers produced healthy calves which lacked antibody in pre-colostral serum. Pathological changes occurred in the foetus in 2 out of 3 pregnant ewes and the ewe produced a healthy lamb which had antibodies to WSL virus in pre-colostral serum.

Unlike the situation in guinea pigs, cattle sera were monospecific for WSL virus in CF tests, but sheep sera cross-reacted with Banzi and yellow fever viruses. Re-infection of the cattle with Banzi, West Nile, Spondweni and Usutu viruses failed to induce marked antibody responses. The results suggest that antibodies to WSL virus in cattle sera from the field can be distinguished from those induced by other flaviviruses by quantitative serologial tests.

3. HAI antibodies to WSL virus were detected in 2648/14395 cattle sera tested over 11 years from 1967 to 1978 in the course of investigation of abortion, infertility and other diseases. Results of quantitative HAI, CF and NT tests with six flaviviruses on 409 selected sera confirmed that infection was due to WSL virus. Serological evidence failed to implicate WSL virus as a cause of abortion in cattle. In a prospective study, abortion occurred in only one out of 21 heifers observed to gain WSL infection during pregnancy in the field, but abortion also occurred in five out of 207 heifers which did not become infected with WSL. No histopathological lesions diagnostic of WSL disease were observed in 1998 specimens from cattle, sheep and goats examined over 44 months prior to October 1972, and WSL virus was isolated once, from the organs of a cow, out of 2106 specimens from cattle sheep and goats tested virologically over six years from October 1972 to September 1978. HAI antibodies to WSL virus were detected in one out of 374 sera from aborted cattle foetuses. It was concluded that WSL virus is not an important cause of disease in cattle, despite widespread occurrence of infection.

In a multi-laboratory trial, minerals-modified glutamate medium (MMGM) was compared with lauryl tryptose lactose broth (LTLB) in the multiple tube method for the enumeration of coliform organisms, including Escherichia coli, in water. Samples of raw and chlorinated waters yielded a total of 2313 positive tube-reactions with MMGM and 2174 with LTLB. These were interpreted either as E. coli; other coliform organisms; or as false positive reactions. The results at first reading (18 or 24 h) and at 48 h have been analysed statistically in terms of (i) most probable numbers of coliform organisms; (ii) positive reactions and their interpretation; and (iii) whether or not the sample yielded any E. coli or other coliform organisms. All three analyses indicated the same trends. For the detection of E. coli in raw waters LTLB was better than MMGM at 18–24 h, but MMGM was better at 48 h with waters containing small numbers of coliform organisms; for raw waters with greater numbers of organisms, both media performed equally well. Analysis of a subset of samples read at both 18 and 24 h indicated that the superiority of LTLB over MMGM with raw waters disappeared by 24 h. For chlorinated waters, LTLB yielded more positive gas reactions at 18–24 h, but fewer of these were E. coli than with MMGM; at 48 h MMGM was clearly better than LTLB for total coliform organisms including E. coli – especially if the numbers were small. MMGM therefore remains the medium of choice for the detection of E. coli as an indicator of faecal contamination of chlorinated drinking water supplies. It is also better for the detection of small numbers of E. coli in other waters.

In a multi-laboratory trial, lauryl tryptose mannitol broth (LTMB) and minerals-modified glutamate medium with added tryptophan (MMGM + T) were compared as single tube tests for the confirmation of Escherichia coli in water; the confirmed results were also compared with the production of gas from minerals-modified glutamate medium without added tryptophan (MMGM). LTMB and MMGM + T gave similar gas and indole results with about 90% of the water samples in most of the laboratories. When compared with the ‘correct’ results as judged by acid and gas production from lactose peptone water and indole from tryptone water, the difference in the rate of false positive reactions between LTMB and MMGM + T was insignificant; but LTMB gave a significantly lower rate of false negative reactions than MMGM + T. Gas production from MMGM without added tryptophan gave significantly higher rates of both false positive and false negative reactions. Lauryl sulphate is therefore a suitable inhibitory surfactant for use in single tubemedia for the confirmation of E. coli, which can be recommended.

In two groups, each containing 56 Pasteurella multocida strains of avian origin, the percentage of multicin-producing strains was 80·4 and 71·4. In two groups containing 46 and 58 isolates tested for sensitivity to multicins, 82·6% and 62·1% respectively were listed as indicators. P. multocida strains producing bacteriocins were active on a range of 1–33 indicators. A preliminary multicin typing system was set up with the use of eight selected indicators; 52% of multicin-producing strains acted on one indicator only. P. multocida serotypes and serologically un-typable strains were subdivided into multicin types.

The spread of insect-borne animal virus diseases is influenced by a number of factors. Hosts migrate, move or are conveyed over long distances: vectors are carried on the wind for varying distances in search of hosts and breeding sites; weather and climate affect hosts and vectors through temperature, moisture and wind. As parasites of host and vector, viruses are carried by animals, birds and insects, and their spread can be correlated with the migration of hosts and the carriage of vectors on winds associated with the movements of the Intertropical Convergence Zone (ITCZ) and warm winds to the north and south of the limits of the ITCZ. The virus is often transmitted from a local cycle to a migratory cycle and back again.

Examples of insect-borne virus diseases and their spread are analysed. Japanese, Murray Valley, Western equine, Eastern equine and St Louis encephalitis represent viruses transmitted by mosquito–bird or pig cycles.

The areas experiencing infection with these viruses can be divided into a number of zones: A, B, C, D, E and F. In zone A there is a continuous cycle of virus in host and vector throughout the year; in zone B, there is an upsurge in the cycle during the wet season, but the cycle continues during the dry season; there is movement of infected vectors between and within zones A and B on the ITCZ and the virus is introduced to zone C by infected vectors on warm winds; persistence may occur in zone C if conditions are right. In zone D, virus is introduced each year by infected vectors on warm winds and the arrival of the virus coincides with the presence of susceptible nestling birds and susceptible piglets. The disappearance of virus occurs at the time when migrating mosquitoes and birds are returning to warmer climates. The virus is introduced to zone E only on occasions every 5–10 years when conditions are suitable. Infected hosts introduced to zone F do not lead to circulation of virus, since the climate is unsuitable for vectors. Zones A, B and C correspond to endemic and zones D and E to epidemic conditions.

Similar zones can be recognized for African horse sickness, bluetongue, Ibaraki disease and bovine ephemeral fever – examples of diseases transmitted in a midge-mammal cycle. In zones A and B viruses are transported by infected midges carried on the wind in association with the movement of ITCZ and undergo cycles in young animals. In these zones and in zone C there is a continual movement of midges on the warm wind between one area and another, colonizing new sites or reinforcing populations of midges already present. Virus is introduced at times into fringe areas (zones D and E) and, as there is little resistance in the host, gives rise to clinical signs of disease. In some areas there is persistence during adverse conditions; in others, the virus is carried back to the endemic zones by infected midges or vectors.

Examples of viruses maintained in a mosquito/biting fly–mammal cycle are Venezuelan equine encephalitis and vesicular stomatitis. These viruses enter a migratory cycle from a local cycle and the vectors in the migratory cycle are carried over long distances on the wind. Further examples of virus spread by movement of vectors include West Nile, Rift Valley fever, yellow fever, epizootic haemorrhagic disease of deer and Akabane viruses.

In devising means of control it is essential to decide the relationship of host, vector and virus and the nature of the zone in which the area to be controlled lies. Because of the continual risk of reintroduction of infected vectors, it is preferable to protect the host by dipping, spraying or by vaccination rather than attempting to eliminate the local population of insects.

The properties of an atypical Brucella strain isolated from lymph node tissue of a cow slaughtered as a brucellosis reactor were examined. The organism was Gram negative and highly pleomorphic, existing as cocci, coccobacilli, rods, branched and irregular forms which stained with fluorescent antibody conjugates prepared against rough and smooth Brucella abortus strains. It produced lecithinase and required at least 15% v/v equine or bovine serum for growth. It did not need supplementary CO2 for growth, produced H2S and was inhibited by brucella dyes and partially by i-erythritol. Growth inhibition or lysis was produced by brucellaphages. The organism was not pathogenic for guinea-pigs or mice but evoked antibodies mainly to rough Brucella antigens.

Growth rates of a mixture of Salmonella serotypes inoculated on beef from a commercial abattoir were measured at chill temperatures. The minimum recorded mean generation times were 8·1 h at 10 °C; 5·2 h at 12·5 °C and 2·9 h at 15 °C. Growth did not occur at 7–8 °C. From these data the maximum extent of growth of Salmonella during storage of meat for different times at chill temperatures was calculated. Criteria for deciding safe handling temperatures for meat are discussed. Maintaining an internal temperature below 10 °C during the boning operation would be sufficient to safeguard public health requirements.

The ability of malachite green/magnesium chloride broth (Rappaport's medium) to isolate salmonellas from 25 ml quantities of sewage-polluted natural water was investigated. Samples were first pre-enriched in buffered peptone water and varying volumes of inoculum from the pre-enrichment culture were inoculated into Rappaport's broth. Inoculum ratios in the range 1:2000 to 1:10 were examined. The inoculum ratio denotes the ratio of the volume of inoculum to the volume of fluid medium into which it is introduced. Optimum results were obtained with the 1:2000 ratio, although the salmonella isolation rate was only slightly less with the 1:500 and 1:100 ratios. The 1:2000 inoculum ratio was obtained with a graduated loop holding approximately 0·005 ml of fluid. Use of a loop for inoculation has advantages in speed of performance and safety of manipulation.

The spread of foot-and-mouth disease during an epidemic in Northumberland in July, August and September 1966 was analysed. Although strong emitters, for example pigs, were not involved, in 18 of the 32 outbreaks spread could be attributed to the airborne route and in another 4, spread by this route was the most likely. Airborne spread was in general between 1 and 8 km but on two occasions as much as 20 km. Other means of spread included movement by animals, people and vehicles and carriage of virus between animals in neighbouring fields. No spread by milk or milk lorries took place. Sheep were involved in 9 farms; on each, lesions older than 48 h were found and it is likely that the sheep were the source of virus for other animals, especially cattle, on the farm and on neighbouring farms. The analysis suggests that control measures such as slaughtering direct contacts, as used in this epidemic, or ring vaccination as well as movement control would be effective in limiting spread.

A regimen of oral non-absorbable prophylactic antibiotics (kanamycin–vanco–mycin–nystatin) was given to nine severely neutropaenic leukaemic patients on cytotoxic therapy (11 courses), in conjunction with isolation procedures. An appreciable decrease in faecal organisms, especially anaerobes, was apparent after 48 h of commencing the course, and most bacteria had disappeared from the stool after five days. There were three episodes of septicaemia, all with enteric organisms, whilst on these antibiotics; one proved fatal. The emergence of resistance to aminoglycosides in faecal flora, notably Klebsiella, in 6/11 courses constituted a major problem in the use of such prophylaxis.

Three selective media were designed for isolation of streptococci from faeces. Samples of faeces from twelve normal adults were suspended and serially diluted in saline or broth saline, and equal volumes of each dilution were spread and incubated on the media. The number of colonies of each different type which developed was counted and one colony of each type was subcultured and identified.

Altogether, streptococci of 13 taxa were found. S. faecium, S. mutans, S. milleri, S. faecalis and S. mitior were each found in over half the samples. Lancefield group B and G streptococci, S. bovis II, an atypical strain of S. bovis I, S. cremoris, S. durans and a dextran-positive strain of S. mitior were each present in 1 or 2 samples. Individual samples contained 2–7 (mean 4·6) streptococcal taxa, and total viable counts of streptococci of 3 × 103−3 × 108 (geometric mean 7 × 105) per g. The significance of these findings is discussed.