The plasmid pSLT is a cryptic plasmid of 60 megadaltons (Md) present in Salmonella typhimurium LT2. We present evidence that it has a characteristic fingerprint when digested with the restriction enzymes PstI and SmaI. Among a representative collection of S. typhimurium isolates it was present in 67% of strains and was widely distributed amongst different phage types (DT) with the exception of DT10 and U285. Furthermore, its prevalence among veterinary isolates was significantly higher than among human isolates. It was not found among any of the 96 strains representative of other salmonella serotypes currently prevalent and thus appears to be serotype-specific.

The distribution of plasmids was studied in a representative collection of salmonella strains which comprised 98 Salmonella typhimurium and 96 other serotypes. Plasmids were detected in 72% of strains (mean 1·3 plasmids/strain) and individual strains harboured between 0 and 7 plasmids. They were more common among S. typhimurium than other serotypes (incidence 92 and 53%; mean 1·9 and 0·8 plasmids/strain respectively). Although a higher proportion of S. typhimurium (33%) were antibiotic-resistant compared to other serotypes (14%) the evidence presented indicated that R-plasmids were not responsible for the difference observed in the number and distribution of plasmids in these strains. These results were discussed in comparison with similar studies of Escherichia coli and other enteric genera.

Restriction enzyme fingerprints were generated from purified plasmid DNA from 324 clinical isolates that belonged to 7 enterobacterial genera and 88 single plasmids in Escherichia coli K12 according to the following strategy.

Purified plasmid DNA was digested with PstI. The number of fragments detected in a 0·8 agarose gel was used to determine which 2 of 6 restriction enzymes including Pstl was most likely to provide a fingerprint comprising sufficient fragments to ensure specificity but sufficiently few to allow easy visual assessment and minimize coincidental matching. When PstI produced > 20 fragments, Eco RI and HindIII were used; when PstI generated < 6 fragments Bsp 1286 and AvaII were used and SmaI was employed when between 6 and 20 fragments were obtained from PstI digests. Using a minimum of 12 fragments from a combination of 2 enzymes as the criterion for characterizing a strain/plasmid, satisfactory 2-enzyme fingerprints were obtained from 87% of the strains and plasmids studied using PstI and no more than two additional enzymes per strain. Of the remaining 54 strains, 51 harboured only small plasmids (< 10 kb) and 3 produced satisfactory fingerprints when digested with a fourth enzyme.

Plasmid profiles have been established for 68 isolates of Staphylococcus aureus from 13 episodes of epidemic spread in hospital wards between 1958 and 1962. Despite the original lack of care in preservation of strains the profiles give, in general, the same epidemiological patterns as were established originally on the basis of phage type, antibiotic sensitivity, ward and date of isolation.

In the Netherlands, case histories of 160 patients aged more than 1 month, with meningitis due to bacteria other than Neisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae were reviewed in order to look for associations between the bacteriological data and the course of disease.

The incidence of such cases was about 0·8/100000/year. Escherichia coli and Listeria monocytogenes each accounted for about 15% of the cases. The case-fatality rate was 18·8% (Gram-negative bacteria, 25%; Gram-positives, 15%) and sequelae occurred in 13·3% of the surviving patients (14 and 13% Gram-negative and Gram-positive, respectively). Hearing loss was the most prevalent sequela (50%). Predisposing factors were present in 70% of patients (69 and 71% respectively), especially in meningitis due to enteric Gram-negative bacteria (except for salmonella) and due to staphylococci.

Surveillance is important because the incidence of meningitis due to these micro-organisms is likely to increase and because the problems in antibiotic treatment have not yet been solved.

In an outbreak of gastroenteritis on board a cruise ship 251 passengers and 51 crew were affected and consulted the ship's surgeon during a 14-day period. There was a significant association between consumption of cabin tap water and reported illness in passengers. Enterotoxigenic Escherichia coli were isolated from passengers and crew and coliforms were found in the main water storage tank. Contamination of inadequately chlorinated water by sewage was the most likely source of infection.

A low level of reported illness and late recognition of the outbreak delayed investigation of what was probably the latest in a series of outbreaks of gastrointestinal illness on board this ship. There is a need for a national surveillance programme which would monitor the extent of illness on board passenger cruise ships as well as a standard approach to the action taken when levels of reported illness rise above a defined level.

Two dairy herds, situated on a sewage farm, were monitored for the presence of salmonellas following outbreaks of Salmonella dublin infection. In addition an S. dublin control scheme, which involved examination of adult animals and calf vaccination, was instigated.

During the period 1975–84, 12 salmonella serotypes and 10 phage types of S. typhimurium were isolated from the cattle and their environment although their presence was seldom associated with disease. Two adult S. dublin excreters were detected but it was concluded that none of the tests employed to examine the adult animals was sensitive enough. The prevalence of disease in the calves was low and although vaccination may have been beneficial it did not eradicate S. dublin infection. Thus S. dublin persisted in adults and calves during the 8-year period but its presence was seldom associated with disease. The results are discussed with regards the disease risk to animals from the agricultural use of sewage sludge and the public health aspects.

One thousand pharyngeal swab specimens were processed for aerobic culture to determine the carriage rate of Gram-negative bacilli (GNB). The isolates were identified and their sensitivity determined to 11 antibacterial drugs following standard techniques.

Similar pharyngeal carriage rates of GNB were found among the various groups of healthy subjects. Patients had higher colonization rates (27%) than healthy subjects (16%). The increase in prevalence of GNB seemed to be associated with underlying diseases and duration of hospitalization.

Klebsiella (36%) was the most frequent genus amongst the 215 isolates of GNB followed by pseudomonas (13%), enterobacter (13%) and acinetobacter (10%). Others were less frequently isolated.

Over 70% of all isolates were resistant to ampicillin (79%) and carbenicillin (72%); 55, 45 and 43% were resistant to cephalothin, tetracycline and streptomycin, respectively. The great majority of the strains were sensitive to the remaining six drugs.

The hospital isolates were more resistant than the non-hospital isolates to most drugs tested. The hospital strains were also more often multiply resistant (89%) than the non-hospital strains (60%). Sixty-five different resistance antibiograms of 1—10 drugs were observed among 191 strains. More varied types of antibiograms were observed among hospital strains.

The high frequency of multiple drug resistance of the isolates is an indication of the extensive use of antibacterial drugs, indicating the need for a policy for judicious use of drugs.

Investigation of 39 JK-type coryneform isolates from patients at a single hospital revealed that 23 possessed plasmids, which formed six groups on restriction endonuclease analysis. Four of the groups were associated with production of similar bacteriocin-like substances, and shared a minimum of 6·4 kilobase pairs of DNA. These plasmids, found in isolates from different patients, provide strong direct evidence that person-to-person transmission of JK bacteria had occurred within the hospital.

The prevalence of nasal colonization and infection with methicillin-resistant Staphylococcus aureus (MRSA) among patients and staff was studied in a section of a Paediatric Surgical Unit in Lisbon between February and July 1985. Nasal colonization was demonstrated in 41% of burned patients, 5% of non-burned patients and 35 % of the nurses. Infection by MRSA occurred in 30% of the burns.

The isolates had identical serological patterns, slight differences on phage typing and were resistant to methicillin, cephalosporins, tetracycline, erythromycin and aminoglycosides. A chloramphenicol resistance plasmid of 3 Md was present in those isolates which were chloramphenicol resistant and a small plasmid of 1·7 Md which coded for constitutive erythromycin resistance was present in many isolates. Gentamicin, tetracycline and inducible erythromycin resistance were chromosomal.

Several reasons for the apparent low virulence of the isolates are discussed. Attempts to control the outbreak by the discharge of colonized or infected patients, improvement of nursing practices and treatment with temporary removal from work of the colonized nurses did not eliminate the organism from the unit.

A multiple-tube technique based on peptone water incubated at 44 °C for 24 h followed by detection of indole was found to be sensitive and specific for the detection of Escherichia coli in oysters and mussels. The method has the advantage of providing rapid results and is both less expensive and less time-consuming than other MPN techniques.

A total of 1203 unselected routine faecal samples from 1006 patients were cultured for Yersinia species by a cold enrichment technique. Seventy-five specimens (6·1%) from 63 patients were culture-positive for Yersinia spp. Fifty-two were Yersinia enterocolitica, 22 Yersinia frederiksenii and 1 Yersinia intermedia. The predominant Y. enterocolitica isolates belonged to biotype 1 – serotype 0:6, 30 or serotype 0:5, 27. Y. frederiksenii strains were non-typable. Forty isolates were recovered from 33 patients with gastroenteritis. During the study period 83 Salmonella spp. from 33 patients, 17 Shigella sonnei from 13 patients and 13 Campylobacter jejuni from 12 patients were cultured. Yersinia spp. was isolated in association with salmonella on three occasions, twice with rotavirus and once each with Shigella sonnei, Campylobacter jejuni and Trichuris trichiura.

Application of a skin lotion to the body after showering greatly reduced the number of bacteria and skin scales dispersed from 10 men and 10 women. This effect lasted for at least 4 h when surgical clothing was worn. The use of a skin lotion to reduce bacterial dispersal could provide a simple and inexpensive alternative to an ultraclean air system or uncomfortable operating clothing during surgery requiring these procedures.

Urinary pathogens isolated from patients in general practice, an antenatal clinic and several hospitals in Liverpool during 1984–5 have been tested for antibiotic sensitivities. The proportion of sensitive organisms varied from antimicrobial to antimicrobial and from institution to institution. Isolates from all institutions showed high rates of sensitivity to cephradine, nalidixic acid and nitrofurantoin, and somewhat lower rates to trimethoprim. Significantly lower sensitivities were found to ampicillin and sulphamethoxazole indicating that neither ampicillin nor a sulphonamide is suitable for initial choice on a ‘best guess’ basis in the situation studied. In general, the organisms derived from the antenatal patients showed the highest rates of sensitivity and those isolated from patients in geriatric hospitals the lowest.

In two trials the efficacy of inactivated vaccines against contagious bovine pleuropneumonia was tested by exposing vaccinated cattle to droplet infection provided by close contact with experimentally infected ‘donors’. Complete protection was given by an extreme form of vaccination in which a heavy suspension of killed Mycoplasma mycoides subsp. mycoides emulsified with Freund's complete adjuvant was given in two large doses. ‘Mouse-protective antibody’ (MPA) was also produced, i.e. serum transferred to mice 2–4 h before intraperitoneal challenge prevented the development of mycoplasmaemia. However, the study did not answer the question ‘Is MPA protective for cattle?’. No protection was given by a milder form of vaccination in which a lighter suspension of killed Hiycoplasmas emulsified with Freund's incomplete adjuvant was given in a comparatively small dose on a single occasion.

Nine serological techniques were compared by monitoring the response to infection with Rift Valley fever (RVF) virus in three sheep. Antibodies were monitored daily for the first 14 days after infection, then weekly and later fortnightly up to week 24. The earliest antibody response was detected in one sheep on day 3 by a plaque reduction neutralization test, and by day 6 antibodies were demonstrable in all three sheep by haemagglutination-inhibition, reversed passive haemagglutination-inhibition, immunodiffusion, indirect immunofluorescence (IF), enzyme-linked immunosorbent assay and neutralization of cytopathic effect in cell cultures. Antibodies were demonstrable by complement fixation on day 8 at the earliest. IF and the two neutralization techniques produced the highest titres, but all tests could be used satisfactorily for the serological diagnosis of RVF. Inactivated antigen could be used for all except the neutralization tests. A radioimmunoassay technique using 125I-labelIed staphylococcal protein A detected antibodies on day 8 at the earliest and produced lower mean titres than some of the other techniques. This was probably because sheep immunoglobulins bind protein A poorly.

Homologous and heterologous haemagglutination-inhibition (HAI), complement- fixation (CF), immunodiffusion (ID) and mouse neutralization tests were performed with the Lunyo (LUN) and a Zimbabwean strainof Rift Valley fever (RVF) virus, the prototype and a South African strain of Arumowot (AMT) virus and prototype strains of Gordil (GOR), Saint-Floris (SAF) and Gabek Forest (GF) viruses, using immune mouseascitic fluids prepared against these viruses. Reactions of identity occurred in all tests between LUN and the Zimbabwean strains of RVF and between the two strains of AMT virus. Otherwise, cross-reactions occurred between all the phleboviruses in HAI tests, while reactions in CF, ID and neutralization testswere monospecific for virus serotypes, except that weak cross-reaction occurred between GOR and SAF viruses in CF and ID tests.

Four sheep infected subcutaneously with the Zimbabwean strain of RVF virus developed transient fever, viraemia, leucopaenia, relative thrombocytopaenia, haemoconcentration and raised serum enzyme levels, which indicated that the sheep had developed necrotic hepatitis. Disseminated focal necrotic hepatitis was confirmed in a sheep killed for examination on day 4 post-infection. The other three sheep recovered uneventfully after only mild depression and anorexia. Groups of three sheep infected with SAF, GOR, AMT and GF viruses had no demonstrable viraemia or other sign of infection or illness, except that the sheep infected with AMT developed mild fever lasting less than 24 h.

Antibody responses were monitored at intervals over a period of 24 weeks in all sheep by homologous and heterologous HAI, CF and cell culture neutralization (CPENT) tests. Homologous antibody responses were marked in the RVF-infected sheep and their sera cross-reacted strongly in HAI tests with antigens of the other viruses. The sera of the RVF-infectes sheep cross-reactes less marledly in CF and CPENT tests. Homologous antibody responses were poor in all the sheep infected with phlebovirused other than RVF, and the cross-reactivity of their sera RVF antigen or virus was negligible. All sheep were challenged with RVF virus 48 weeks after their initial infection. The sheep which had originally been infected with RVF virus were immune and developed neither fever nor viracmia. All other sheep developed fever, viraemia and antibodies to RVF virus.

It was concluded that the African phleboviruses, other than RVF, are unlikely to cause disease in livestock ot to induce antibodies which could cause confusion in the diagnosis of RVF.

Influenza surveillance in the UK between the years 1982 and 1985 has demonstrated the regular winter appearance of influenza A virus of both H1N1 and H3N2 subtypes and influenza B.

Their antigenic diversity is described and correlated with the national statistics for morbidity and mortality for influenza.

One unexpected finding has been that despite the wide circulation of influenza viruses there has been a continuation of winters without significant increases in influenza deaths or morbidity. A previous report of influenza surveillance (Pereira & Chakraverty, 1982) noted an already unusual series of three consecutive winters with this pattern. This report records a further 4 years bringing a total of seven successive winters without evidence of epidemics of severe disease associated with influenza viruses, as indicated by the national UK statistics.

To determine the pathogenic potential of the vesiculoviruses Isfahan Chandipura for domestic animals, two ponies, two steers, three and three pigs were inoculated with each virus intradermally in the tongue or, in the case of the pigs, in the snout, heel and coronary band The Ponies were also inoculated intradcrmally in the right commissure o the mouth Animals inoculated with each virus were housed in one room and allowed to mingle freely with an equal number of uninoculated contact animals of each species.

Clinical signs of infection, consisting of ulcers at the inoculation sites, were observed in the Chandipura study in two inoculated ponies, one inoculated steer and one inoculated goat. No elevated temperature was observed. Virus was isolated from the ulcerated tongue tissue but not from serial blood samples, oesophagcal-pharyngeal mucus samples, or from the tissues which were collected at necropsy. Precipitating antibody was not detected by the immunoelectro osmophoresis(IEOP) test in any of the pre- or post-serum samples except from two inoculated sheep at 29 days post-inoculation (D.P.I.). Low levels of neutralizing activity were dtedted in pre-inoculation serum from all steers, pigs, contact sheep, and one contact goa. By 15 D.P.I. all inoculated animals and contact ponies and steers exhibited increased neutralizing antibody titres.

In studies with the Isfahan virus, lesions developed only at the inoculation sites in the two ponies, and the virus was isolated. No virus was isolated from any blood, oesophageal-pharyngeal mucus samples or tisues collected at necropsy. All pre-inoculation sera were negative for neutralizing and precipitating antibodies. By 14 D.P.I. all inoculated animals exhibited neutralizing antibody, while all the contacts remained negative. The IEOP test remained negative for all animals throughout the experiment. A subpassage of a suspension of Isfahan-infected tongue tissue injected in to ponies and steers also yielded only firm swellings of lesser extent than the original reaction at the inoculation sites.

With both viruses, lethal infections were produced by intraacranial or intraperitoneal inoculation of day-old mice and hamsters, and by allantoic inoculation of embryonating chicken eggs. Adult mice, hamsters, guinea-pigs and rabbits produced serum antibodies but lacked clinical signs.

Thirty-six strains of haemorrhagic fever with renal syndrome (HFRS) virus were isolated from patients and a number of host animals in various areas in China. They were analysed by an immunofluorescence test (IFAT) using 10 monoclonal antibodies (McAbs) specific for the HFRS virus; antigenic differences among the strains have been demonstrated. The HFRS virus strainsrevealed nine different reactions with the McAbs, showing that there are at least nine different antigenic determinants including group-, type- and strain-specific. Analysis of the results shows that antigenic differences among the HFRS virus strains are mainly related to differences in the host animals.