Enterococcisensu stricto form part of the normal gut flora (1) and may be found in the mouth, vagina and anterior urethra (2). They are opportunist pathogens which can cause serious infection including endocarditis. Nosocomial enterococcal infection appears to be increasing both in the UK (Public Health Laboratory Service [PHLS] Communicable Disease Surveillance Centre [CDSC], unpublished) and the USA (3) and to correspond to usage of broad spectrum β-lactam antimicrobial agents (4−7) and invasive surgical devices (8, 9). At the same time, the incidence of enterococci resistant or tolerant to previously commonly employed antimicrobial agents or their synergistic combinations is increasing and is compromising therapy of serious enterococcal infection. Strains of enterococci with high-level resistance to streptomycin and kanamycin (minimal inhibitory concentrations [MICs] > 2000 mg/L) were first reported in 1970 (10, 11) and rapidly became widespread (8, 12−14).

Two small flocks of egg-laying hens, naturally infected with Salmonella enteritidis. were housed in individual cages so that their eggs could be identified. During a longitudinal study where the contents of 1119 eggs were examined, 11 were positive for S. enteritidis. One isolate was phage type (PT) 33 the others were PT4. The production of infected eggs was clustered though intermittent. The positive eggs. which were produced by 10 of the 35 hens, were all found to contain fewer than 10 salmonellas. Some birds were also apparently carrying S. hadar PT14 as this organism was isolated from the contents of six Cracked eggs.

In February 1989 the largest reported outbreak to date in the United Kingdom of Salmonella enteritidis phage type 4 (PT4) infection occurred following a wedding reception at a hotel. One hundred and seventy-three people met the case definition of illness of whom 118 had the organism isolated from their stools. A further 17 were found to be S. enteritidis PT4 positive, but were asymptomatic. Lightly-cooked, egg-based sauces were the epidemiologically proven vehicles of infection. Investigations showed this outbreak to be the first to implicate imported European eggs as the source of infection. An unusual feature of this outbreak was a reported incubation period of less than 3 h for some of the confirmed cases of salmonellosis.

An index of discrimination was used to assess the discriminatory power of biotyping for salmonellas of selected serotypes. Three collections of phage-typed strains of Salmonella typhimurium and Salmonella paratyphi B, previously tested for biotype, were examined. The results established that the discrimination index was high when full biotyping alone was used for differentiation of strains from international series. When biotyping was combined with phage typing, the discrimination achieved was greater than for either method used alone. These findings confirm that biotyping affords excellent discrimination of strains of S. typhimurium and S. paratyphi B and indicate that the results obtained from biotyping can be interpreted with confidence.

An outbreak of Salmonella paratyphi B infection in the UK associated with a fish-and-chip shop is reported. The source of infection for the first three cases was believed to be a food handler who was infected overseas 6 years earlier. His wife whose faeces and urine were originally culture negative continued to run the shop but subsequently her faeces became positive on one occasion. She was considered to have been the source of two further cases, and secondary household spread of infection from these two cases resulted in one symptomatic and two asymptomatic infections. A second household contact of the proprietor also became a faecal excretor 2 months later. We recommend that food handlers living in households or in intimate contact with eases or carriers of S. paratyphi B should be put off work until all household contacts cease excreting the organism.

Cheese made from sheep milk was implicated in food-poisoning incidents in December 1984 and January 1985. Bacteriological examination of batches of cheese failed to reveal a viable pathogen but enterotoxin A produced by Staphylococcus aureus was present. This was the first time that enterotoxin was detected in a food produced in the UK which was associated with poisoning and from which viable Staph. aureus could not be isolated. Subsequent detailed examination of milk, yoghurt and cheese from the same producer revealed that contamination with Staph. aureus was associated with post-infection carriage as well as clinical illness in ewes on the farm. Strains producing enterotoxin A were still intermittently present in the bulk milk used for cheese production nearly 2 years afterwards, apparently in the absence of clinical illness in the sheep. The possible effects of heat treatment are discussed. Any changes in legislation should cover all non-human mammalian milk used for human consumption.

Since 1985. nosocomial infections have been frequently treated with a new fluoroquinolone, pefloxacin, at the Cardiological and Neurological Hospital in Lyon. From 1986 to 1988. the incidence of resistance of clinical Staphylococcus epidermidis strains to pefloxacin increased from 31 to 57%. Dispersion of these resistant strains in the hospital environment was recognized when they were detected on 22% of staff members' fingers (139 samples were investigated) and in 28% of the environmental samples (180 were investigated). There was an association between carriage rate and work place. Most of the pefloxacin-resistant S. epidermidis were resistant to oxacillin, gentamicin. erythromycin, cotrimoxazole and fosfomycin. Intensive use of pefloxacin selected multiresistant S. epidermidis which became ubiquitous in the hospital environment.

The study was conducted to determine whether pre-enrichment would increase sensitivity of detecting Streptococcus (Str.) aqalactiae, Staphylococcus (S.) aureus, and mycoplasma in bovine milk. Two procedures were followed, one involving direct inoculation of milk on bovine blood agar, and the other involving preenrichment in broth followed by inoculation on agar. Logistic regression was used to predict the probability of isolation as a function of culture procedure and two additional covariates. the California Mastitis Test (CMT) score of the milk and the type of sample (indicating sample storage temperature and herd mastitis status). A total of 13778 milk samples was cultured for each of the three bacteria. By using results of both direct inoculation and pre-enrichment, the probability of isolation compared to use of direct inoculation only and adjusted for effects of other variables was increased 3·6-fold for Str. agalactiae, 1·6-fold for S. aureus and 1·7- fold for mycoplasma. The probability of isolation for all three bacteria increased as the CMT score increased. For Str. agalactiae, there was a statistical interaction predicting that enrichment improved the odds of isolation more from milk with high CMT scores than from milk with low scores. Results indicate that preenrichment can substantially increase the sensitivity of bacteriological screening of dairy cows for mastitis caused by Str. agalactiae, S. aureus, and mycoplasma.

The possible relationship between the results of a microbiological sampling programme and visual inspections carried out in local food-manufacturing premises was examined. Using five main parameters—overall appearance, personal hygiene, risk of contamination, temperature control, and training and education –a visual inspection rating score was established for each of the premises. A variety of high-risk processed foods, and specimens from hands, wiping cloths and environmental swabs were examined. The results from two study Periods indicated that there was an overall poor agreement between microbiological results and inspection ratings. On its own, neither sampling nor visual assessment reliably monitored the performance of the premises. A combined approach, using selective microbiological examination to support a system of standardized inspections, is suggested for monitoring food hygiene standards in premises selling high-risk foods

The investigation of plasmid similarity is an important component in the surveillance of antimicrobial resistance and in the detection of epidemic plasmids. The use of restriction endonucleases in the classification of transferable, multiply-resistant plasmids from faecal Enterobacteriaceae isolated at the Children's Emergency Hospital, Khartoum was investigated. Twenty-four transconjugant plasmids, coding for 11 different resistance patterns, each of molecular weight 62 MDa. were studied using four restriction enzymes; Pst I, Eco R I, Hind III and Ara II. Fifteen different digest profiles were obtained. Restriction profiles discriminated between plasmids with differing resistance patterns and demonstrated homology of plasmids with common resistance patterns. Restriction endonuclease digest patterns provide a potentially rapid and reproducible method of plasmid classification, that could contribute towards surveillance systems in tropical countries with a high prevalence of antimicrobial resistance.

The membrane-filter assay, GM1-ELISA, and DNA-DNA hybridization assay, were used to detect enterotoxigenic Escherichia coli (ETEC) in samples of water, weaning food, food preparation surface swabs, fingerprints of mothers, and the fingerprints and stools of children under 5 years of age, in 20 households in a Malaysian village. Weaning food and environmental samples were frequently contaminated by faecal coliforms, including ETEC. The membrane-filter assay detected and enumerated faecal coliforms and LT-ETEC in all types of water and weaning food samples. Highest concentrations of faecal coliforms and LT-ETEC were found in weaning food, followed by well-water, stored water and stored drinking water. The GM1-ELISA detected LT-ETEC in weaning food, food preparation surfaces, fingerprints and stool samples. The DNA-DNA hybridization assay detected a larger proportion of STa2-ETEC than the other toxotypes, either singly or in combination. All the assays in combination detected the presence of ETEC in all types of samples on at least one occasion in each household. It was not possible to classify households as consistently more or less contaminated with ETEC. On individual occasions it was possible to show a significant association of the presence of LT-ETEC between the fingerprints of children and their stools, fingerprints of mothers and children, and weaning food and the stools of the child consuming the food.

Eight hundred and sixty-four Escherichia coli isolates from workers at the University of Ibadan Teaching and Research Poultry Farm, and 216 isolates from poultry attendants at a commercial poultry farm in the city were found to be resistant to streptomycin, sulphafurazole and tetracycline. In contrast, all 576 and 288 E. coli isolates from village fowls and from villagers respectively were sensitive to these drugs. Isolates from birds in a modern university poultry unit (3744) exhibited the same resistance patterns as those isolated from workers who were in direct contact with the birds. No nalidixic acid-resistant E. coli was isolated from farm workers prior to their assignment to the experimental pen. Following experimental oral infection of birds with E. coli K12 J5 NA Lac, the organism was recovered from the workers who manned the experimental pen. Neither before nor after the experimental infection was any nalidixic acid resistant E. coli isolated from workers who manned the pen from which birds used in the experiment were selected. Similarly, no drug resistant organisms were isolated from workers outside the poultry unit of the university or commercial farm. The MIC of the drugs against the avian and human E. coli isolates at the university and commercial poultry farms were similar.

Replicate faecal samples from healthy individual pigs and cows were examined for the presence of Aeromonas sp. over a 12-month period. Aeromonads were found to be minor components of the faecal flora, only 8·8 % of 520 samples from pigs and 4·6% of 481 samples from cows proving positive. Isolation rates in both groups of animals were seasonal. A. hydrophila (62% of the isolates) was the predominant species in cows, followed by A. caviae (32%) and A. sobria (15%). This pattern was also recorded in the natural waters that the animals drank from during the period when the faecal carriage rate was at its highest. In pigs, A. caviae (59%) was more common than A. hydrophila (41%). A. sobria was not found in any of the pig-associated samples. It seems that cattle acquire their faecal aeromonads from drinking water. The source of the organisms in pigs is less clear.

A 10 min education programme was developed which, if effective in changing the behaviour of pregnant women, would eliminate or greatly reduce the risk of congenital toxoplasmosis. It was taught in 26 randomly selected (case) prenatal classes offered to women early in their pregnancy. The remaining 26 (control) classes received routine class material which did not mention toxoplasmosis. A questionnaire was administered to all women prior to this early class (pre-test) and again after the last prenatal class, held just prior to delivery (post-test). Changes in pet, food and personal hygiene behaviour between the pre- and post-test were determined and a score calculated by adding points for change towards those behaviours recommended in the programme and subtracting points for change in the opposite direction. Cat owners in case classes had a significantly higher score in pet hygiene behaviour than those in control classes (P < 0·05). No significant difference was found between the food or personal hygiene scores of women in case and control classes, possibly because of low power. However, although behaviours did not differ on the pre-test, women in case classes had significantly better cooking methods for roast beef and hamburger on the post-test (P < 0·05 and P < 0·01 respectively). It is concluded that this programme is effective and should be offered to all women in order to reduce congenital toxoplasmosis incidence.

A new epidemiologic typing method based on electrophoresis of esterases had been developed for differentiating between clinical isolates of Branhamella catarrhalis. Twenty-two epidemiologically significant strains obtained from three Chest Units, a Paediatric Intensive Care Unit and a Paediatric Unit were compared with 54 randomly selected strains and 4 reference strains, including the species type strain, ATCC 25238. Thirty-four distinct zymotypes were characterized by polyacrylamide-agarose gel electrophoresis of the 80 strains. One infrequent zymotype was found in 2 neonates and another in 2 adults with nosocomial bronchopulmonary infections, suggesting the nosocomial spread of 2 outbreak strains of B. catarrhalis. A more frequent zymotype was isolated from 3 neonates with nosocomial bronchopulmonary infection and from 2 children with nosocomial rhinopharyngitis. The remaining 12 epidemiologically significant strains were of varied zymotypes. This work demonstrates that esterase electrophoresis is a suitable, readily reproducible, stable typing system applicable to the wide range of strains found in B. catarrhalis nosocomial infections.

Three hundred and fifty-eight stool and 131 sputum specimens from 40 cystic fibrosis (CF) patients and 100 toilet sinks were investigated for occurrence of Pseudomonas aeruginosa; 67% (21/31) of the patients with chronic P. aeruginosa lung infections carried the organism repeatedly in the stool but the organism was found only once in the stools of nine uninfected patients. P. aeruginosa stool carriage was correlated to high P. aeruginosa numbers in patients' sputa. Typing of P. aeruginosa with a DNA probe showed identity of sputum and stool strains. Seven patients repeatedly carried additional stool strains, not found in the sputum, suggesting intestinal colonization. No differences were seen in the clinical state of patients with P. aeruginosa-negative stool samples and patients with positive stool samples. Toilets in households of P. aeruginosa-infected CF patients were significantly more often contaminated with P. aeruginosa (42%) than toilets in households of non-infected CF patients (20%; P < 0·03). The study shows that P. aeruginosa-infected CF patients may harbour the organisms also in the intestinal tract, and may spread the bacteria into toilets.

A survey was made of serotype association and multiple antibiotic resistance in strains of Pseudomonas aeruginosa in Europe. Of 208 epidemiologically distinct strains from 16 laboratories in 10 countries, 48 were resistant to carbenicillin (MIC > 128 μg/ml) and gentamicin (MIC > 4 μ g/ml), and 12 of these strains were of serotvpe O 12. Representative O 12 strains from different countries were compared with two multiresistant O 12 strains isolated 5 years apart, from a British burns unit and the antibiotic sensitive serotype reference strain. All O 12 strains were similar by phage and pyocin typing but lysogenic phage profiles indicated that two strains (the later burns isolate and the serotype strain) were distinct from the others. Electrophoretic characterization of outer membrane proteins, lipopoly- saccharides and esterase enzymes corroborated the relationship of the strains and restriction fragment length polymorphism of DNA fragments hybridized with a cDNA probe copy of rRNA from P. aeruginosa provided further proof of their relatedness. We propose that the uniformity of characters of multiresistant O 12 P. aeruginosa in Europe is suggestive of a common origin for the strains.

Graded doses of live and heat-killed cells of Campylobacter jejuni were injected into the yolk-sac of 5-day-old chick embryos, and the 50% lethal dose (LD50) was determined 7 days later. A strain dependent virulence was seen. In the diluted series of cultures the LD50 values for live campylobacter ranged from 106 c.f.u. beyond the last dilution showing growth, that is to less than one organism per embryo. When the 22 strains were tested as heat-killed cells, the chick embryo LD50 values retained the same relative order of toxicity obtained with viable cells, but the LD50 values were increased by + 1 to + 4 log units. Heat-killed cells from strains known to be invasive, but non-toxigenic, were still lethal for the embryos, suggesting that viability was not solely necessary for virulence. Semi-pure lipopolysaccharide from a non-virulent strain of C. jejuni was not toxic to the embryos, but semi-pure and ultracentrifuge-purified lipopolysaccharide from the most lethal campylobacter strains gave LD50 values in the order of 3·0 μg lipopolysaccharide per ml (0·6 µg per embryo) in the yolk-sac assay. No relationship between serotype and lethality was seen. Injection into the yolk-sac appears to be an easy, rapid and reproducible in vivo assay of the virulence of C. jejuni.

The diphtheria immunity status was determined with the passive haemagglutination technique in 503 sera of 10–90-year-old persons from Warsaw and Olsztyn Provinces. Donors of sera were students, teachers, pregnant women, employees of industry and medical service. The immunity was highest (90% of titers 0·1 lU/ml or higher) in persons below 20 years of age and in persons above 60 years of age (55%). Between these two groups, gaps in immunity exist, the proportion of those immune varying from 36–50% in the 20- 60-year-old groups.

The outer membrane proteins of 38 Yersinia pestis isolates from all known plague foci of north-east Brazil were analysed by SDS-PAGE. Approximately 20 bands were consistently found in all strains analysed and 11 were selected for comparative studies. Although qualitative differences among the electrophoretic profiles of outer membrane proteins of wild Y. pestis isolates were not observed, quantitative alterations were clearly noted for most of these proteins. No particular quantitative alteration of the electrophoretic profile of outer membrane proteins could be associated with the period of isolation and geographic origin of the isolates. The 64 kDa outer membrane protein was significantly expressed in higher amounts among Y. pestis strains isolated from a recent plague outbreak. The possible use of electrophoretic profiles of outer membrane proteins of wild Y. pestis isolates as a tool for epidemiological studies and for the analysis of virulence determinants is discussed.