Sixty-nine Staphylococcus aureus strains. 39 of which produced staphylococcal enterotoxin B (SEB + ) and 14 of which were associated with toxic shock (TS + ), were studied using the following markers: serotyping, phage typing, antibiotyping. ribotyping, zymotyping and pulsed-field electrophoresis typing. Analysis of the results showed that the enterotoxin B producing strains were derived from at least three clones: the first two consisted of methicillin-susceptible strains, while the third included the methicillin-resistant (MRSA) strains. TS+ strains of non-genital origin appeared to be distributed between the three clones, with no specific characters.

Aerosol infection (AI) of Porton outbreak mice with Listeria species, exhibiting varying degrees of virulence, was compared with gastric intubation (GI) on the basis of numbers of deaths. 50% lethal dose (LD50) and pattern of listerial infection. The AI route appeared to be more sensitive, efficient and consistent than GI in that it required 105 fewer micro-organisms to obtain infection and death then ensued within 4 days, with GI deaths usually occurring on day 7. All the virulent strains tested caused 100% mortality by AI, while virulent and avirulent strains were indistinguishable by GI. Bacterial counts in the livers and spleens of infected mice were consistent with the relative virulence of the infectious agent using AI but not in GI mice. There were higher numbers of micro-organisms and more widespread lesions in the organs of AI mice than in GI. Results indicate that AI is an accurate in vivo indicator of virulence in listeria and using AI, bacterial counts in the liver and spleen could replace LD50 tests, thereby reducing the number of animals required for in vivo virulence testing.

In recent years, methicillin–resistant Staphylococcus aureus (MRSA) isolates in Spain have increased dramatically; in 1986 there were only 1.2% MRSA amongst all nosocomial Staphylococcus aureus (SA) isolates, by 1989 this percentage had risen to 44% in some hospital causing a very serious epidemic situation in the country. We have characterized these isolates by direct, reverse and Fisk phage typing and we have also looked for an additional local set of phages to help us to differentiate these strains. We have been able to differentiate an epidemic strain from other MRSA strains which cause sporadic hospital outbreaks, and we have also distinguished between some variants of the epidemic strain.

The investigation, epidemiology, and effectiveness of control procedures during an outbreak of Legionnaires' disease involving three immunosuppressed patients are described. The source of infection appeared to be a network of fire hydrant spurs connected directly to the incoming hospital mains water supply. Removal of these hydrants considerably reduced, but failed to eliminate, contamination of water storage facilities. As an emergency control procedure the incoming mains water was chlorinated continuously. Additional modifications to improve temperature regulation and reduce stagnation also failed to eliminate the legionellae.

A perspex test-rig was constructed to model the pre-existing hospital water supply and storage system. This showed that through the hydraulic mechanism known as ‘temperature buoyancy’, contaminated water could be efficiently and quickly exchanged between a stagnant spur pipe and its mains supply. Contamination of hospital storage tanks from such sources has not previously been considered a risk factor for Legionnaires' disease. We recommend that hospital water storage tanks are supplied by a dedicated mains pipe without spurs.

Leptospirosis is a worldwide zoonotic disease. In the present investigation, a total of 89 human sera from a flood prone district of Bangladesh was screened by a one-point microscapsule agglutination test (MCAT). MCAT-positive and-doubtful sera were further tested by microscopic agglutination test (MAT) against 16 reference serovars of Leptospira interrogans. and the antibody titres determined. In MCAT, 34 sera were positive and 22 were doubtful. Among those positive and doubtful sera. 33 and 20. respectively were tested by MAT. Thirty-four out of 53 MCAT-screened samples were MAT-positive. The titres ranged from 20 to 1600 with antibodies to serovars copenhageni, australis, cynopteri and icterohaemorrhagiae being the most prevalent. Eleven MCAT-positive samples failed to react with any strains used by MAT, suggesting the presence of new or untested serovars. Among the MAT-positive samples, the presence of antibody against two or more serovars was more common than that of a single serovar. The present study suggests that rural people in Bangladesh are at high risk to leptospiral infection.

Non-specific and specific mechanisms of adherence have been examined in two collections of methicillin–resistant Staphylococcus aureus (MRSA). Determination of hydrophobicity by salt aggregation, hydrophobicity indices and of adherence to the extra–cellular matrix proteins fibronectin, vitronectin, laminin and collagen type 1 have failed to reveal any correlation with phage-type, plasmid profile or antibiogram. Further, the strain collections, made over a period of years in two countries, differ markedly in their adherence characteristics; MRSA are heterogeneous in this respect. Such heterogeneity may explain the polarization of views on the epidemicity or ‘virulence’ of MRSA. With the exception of adherence to collagen a small group of methicillin sensitive S. aureus had characteristics intermediate between the two groups of MRSA.

During the period 1961–91 a total of 567 635 strains of Staphylococcus aureus from hospitalized patients in Denmark have been characterized according to their antibiotic resistance, site of isolation and phage type. Strains of phage group II (typed by the phages 3A, 3C, 55 and 71) have been analysed further. The occurrence of group II strains was relatively constant (approximately 16%) from 1961 until 1983. Since then the frequency of group II strains increased; in 1991 they accounted for 22.7% of all S. aureus strains isolated. Strains of group II can, on the basis of their phage types, be divided in four subgroups: 3A, 71, 71 + and the ‘rest of group II’. Furthermore, within these groups strains may differ from one another in respect to their sensitivity to phages.

The increased isolation of group II strains during recent years was because of an increase in strains of subgroups 71 + and the ‘rest of group II strains’. In 1991 these two subgroups accounted for 89.7% of all group II strains. Furthermore, an increasing number of group II strains, 71.4% in 1991, was typable only at RTD × 100. The increase in the number of group II strains was even throughout Denmark. All four subgroups of group II have, during the observation period, become more frequently resistant to penicillin and/or tetracycline. Strains typed at 100 × RTD of subgroup 71 + and the ‘rest of group II’ are more frequently antibiotic resistant than the rest of the group II strains.

Strains of the increasing subgroups occurred most often in abscesses.

This case control study investigated environmental factors in 74 confirmed cases of meningococcal disease (MD). In children aged under 5, passive smoking in the home (30 or more cigarettes daily) was associated with an odds ratio (OR) of 7.5 (95% confidence interval (CI) 1.46–38.66). ORs increased both with the numbers of cigarettes smoked and with the number of smokers in the household, suggesting a dose–response relationship. MD in this age group was also significantly associated with household overcrowding (more than 1.5 persons per room) (OR 6.0, 95% CI 1.10–32.8), with kisses on the mouth with 4 or more contacts in the previous 2 weeks (OR 2.46, 95% CI 1.09–5.56), with exposure to dust from plaster, brick or stone in the previous 2 weeks (OR 2.24, 95% CI 1.07–4.65); and with changes in residence (OR 3.0, 95% CI 1.0–8.99), marital arguments (OR 3.0, 95 % CI 1.26–7.17) and legal disputes in the previous 6 months (OR 3.10, 95% CI 1.24–7.78). These associations were independent of social class. Public health measures to lower the prevalence of cigarette smoking by parents of young children may reduce the incidence of MD. The influence of building dust and stressful life events merits further investigation.

Ribotyping, with homologous or heterologous (Escherichia coli) r–RNA, of the propagating strains for phages of the international set for strains of Staphylococcus aureus of human origin was undertaken to determine the discrimination of this typing method. Ribotyping could distinguish between strains of different phage groups, but could not distinguish between seven phage group III strains of different phage type. Ribotyping may be a useful adjunct to phage typing in S. aureus but is unlikely to replace it as the primary method of epidemiological typing.

Borrelia burgdorferi isolates were obtained from Ixodes ricinus from three sites in Switzerland. They were examined by SDS-PAGE and immunoblotting. The phenotypes, in respect of three outer surface proteins (Osp), differed between the sites of collection. In site 1, most isolates had an OspA of 31 kDa and an OspB of 34 kDa: in site 2, isolates presenting an OspA of 33 kDa dominated and in site 3, the isolates with an OspA of 32 kDa and an OspB of 35 kDa were most frequent. This distribution differed significantly. About half of the isolates from sites 1 and 3 reacted with anti-OspA monoclonal antibody H5332 compared to 29% from site 2. Site 1 isolates reacted significantly more frequently (81 %) with another anti-OspA monoclonal antibody LA-31 than isolates from site 3 (P < 0·0001). These findings have implications for the epidemiology of Lyme borreliosis, for the further development of serodiagnostic reagents and for the development of a vaccine.

Two hundred and eighteen nosocomial cases of Legionnaires' disease with 68 deaths were reported to the National Surveillance Scheme for Legionnaires Disease between 1980 and 1992, representing 15% of the reported infections acquired in England and Wales. Twenty–two nosocomial outbreaks accounted for 135 (62%) of these cases, the remainder occurring as single cases either in hospitals where other single cases or outbreaks had been reported in different years or as ‘sporadic’ cases in hospitals from which no other cases were reported. A clinical history prior to onset of Legionnaires' disease was available for 124 patients, 61 of whom had undergone recent transplant therapy or were immunosuppressed for other reasons.

Sixty cases (27%) were diagnosed by culture of the organism and isolates from 56 patients were typed; 25 (42%) were non L. pneumophila serogroup 1 infections.

Methods for prevention and control of nosocomial outbreaks are discussed, in particular the susceptibility to Legionnaires' disease of certain groups of hospital patients.

Thin-layer chromatography (TLC) was compared with seroagglutination for subtyping of Mycobacterium avium complex (MAC) bacteria. Seventy-five significant MAC isolates from patients with AIDS were typed by both methods and 36 isolates, judged to be clinically non-significant, were examined by TLC only. Overall. 75% of isolates tested were typable by seroagglutination and 91% by TLC; the results correlated between the two except for minor discrepancies.

Serovars 1, 8 and 21 and mixed serovars 1–21 and 1–8–21 were common among isolates from AIDS patients and together represented 83 % of isolates compared with only 36% in the non-significant group (odds ratio 8·4; 95% confidence interval 3·4–23·3). This difference remained significant after exclusion of serovar 41 (M. scrofulaceum), which was the commonest isolate (28%) in the non-significant group but was not isolated from patients with AIDS.

TLC is useful to supplement seroagglutination for subtyping of MAC. Further study is required to determine whether apparent differences between isolates from patients with AIDS and from other sources reflect differences in virulence or in environmental prevalence of MAC subtypes.

This report describes a double outbreak of staphylococcal scalded skin syndrome (SSSS) in which two distinct tetracycline-resistant strains of Staphylococcus aureus producing different exfoliative toxins were involved. In the first phase the daytime staff of the delivery unit and eczematous skin conditions in midwives were implicated as the probable source. In the second phase a source within a post-natal ward was suggested with local cross-infection. In the final phase both sources were epidemiologically linked to cases of SSSS. Because early discharge was the policy of the unit many cases presented in the community rather than in the hospital.

Confirmation of epidemiological findings was provided by additional laboratory studies. Two distinct strains of S. aureus could be defined, differing in phage-typing patterns, the exfoliative toxin produced, plasmid profile, cadmium resistance and bacteriocin production. Strict care in hand washing with a chlorhexidine-containing detergent was an important control measure.

To investigate the joint association of patient and strain characteristics with the outcome of meningococcal disease (MD), data were collected on 563 consecutive cases of MD reported between 1989 and 1990 in The Netherlands. The meningococcal isolates were characterized with regard to their surface characteristics. Sequelae occurred in 8.5% of the patients, and were only associated with the presence of bacteraemia. The case-fatality rate was 7.7%. Infants aged ≤ 5 months and patients in the age-groups of 10–19 years and ≥ 50 years had an increased risk for a fatal outcome compared with children from 6 months to 9 years old (Odds Ratios [ORs]: 5.1, 3.4 and 9.8, respectively). The OR for females versus males was 2.3. The ORs for patients with bacteraemia, or a combination of bacteraemia and meningitis, compared with meningitic patients, were 2.3 and 3.1. Meningococcal strain characteristics did not influence the case-fatality rate substantially. In conclusion, host factors were found to be determinants for a fatal outcome of MD in The Netherlands from 1989 to 1990.

Sera from 83 patients with campylobacter gastroenteritis were examined for the presence of legionella antibodies by indirect immunofluorescence. Twenty–one patients (25%) had positive titres (≥ 16) including 11 patients with titres of ≥ 128. Legionella seropositivity persisted in 5 of 9 patients (55%) studied for 6–9 months. Campylobacter isolates were serotyped by the Penner scheme. Isolates associated with legionella seropositivity included Penner types 1, 2 and 4, the common endemic serotypes in England. Campylobacter blocking fluids were prepared from a range of Penner reference strains. The blocking fluid prepared from Penner type 11 was the most efficient at inhibiting the false-positive legionella titres. Using this absorption step legionella titres were inhibited from 24 of 26 patients (92%) with campylobacter but not from 8 patients with cultureproven legionnaires' disease. We recommend that this method is incorporated into routine diagnostic legionella serology in order to eliminate false-positive reactions due to campylobacter.

Thirty-two Tenckhoff catheters retrieved from continuous ambulatory peritoneal dialysis patients with a history of peritonitis were examined for microbial biofilm. Confocal laser scanning microscopy was successfully employed to visualize bacteria in biofilm occluded from view by scanning electron microscopy. Occluded but viable microbial biofilm was associated with 17 (81%) catheters from patients free from infection following renal transplant. Mixed isolate biofilm with two or more isolates of coagulase-negative staphylococci or Staphylococcus aureus was found on 41% of these catheters. Clearly visible viable biofilm consisting exclusively of Pseudomonas aeruginosa occurred on all four catheters removed due to recurrent peritonitis. Five (71%) catheters retrieved from patients transferred to haemodialysis had viable biofilm. Antibiotic sensitivities of the biofilm isolates were similar in profile to those reported for non-biofilm isolates from infected dialysate. Persistence of catheter biofilm despite direct contact with therapeutic levels of antibiotics in peritoneal dialysate requires that attention be directed towards improving antibiotic efficacy against peritonitis-causing bacteria in biofilm form.

The conjunctival sacs of 100 healthy adult ewes and 20 lambs and 76 adult ewes affected by ovine keratoconjunctivitis (OKC) were examined microbiologically. Branhamella ovis was the only organism regularly isolated from healthy ewes and lambs. Mycoplasma conjunctivae was isolated from the conjunctival sac of nine ewes but its occurrence was not associated with clinical signs of ovine keratoconjunctivitis. Mycoplasma conjunctivae and Branhamella ovis were isolated significantly more often from eyes affected by OKC than from unaffected eyes.

We studied the relationship between IgA anti–campylobacter flagellin antibodies in breast milk samples and protection of breastfed infants living in a rural Mexican village from campylobacter infection. There were fewer episodes of campylobacter infection (symptomatic and asymptomatic combined) in infants breastfed with milk containing specific anti-flagellin antibodies (1.2/child/year, 95% CI 0.6–1.8) versus non–breastfed children (3.3/child/year, 95% CI 1.8–4.8; P < 0.01). Infants breastfed with milk that was antiflagellin antibody negative by ELISA also had fewer episodes of infection compared with non-breastfed children, but the difference did not reach statistical significance (1.8/child/year, 95% CI 0.7–3.0 versus 3.3/child/year, 95% CI 1.8–4.8, P > 0.05). Breastfeeding has a protective effect against campylobacter infection and is associated with the presence of specific antibodies directed against campylobacter flagellin.

Aminoglycoside resistance patterns of 147 Serratia marcescens strains of clinical origin were studied. All strains analysed belonged to three different bacterial populations. The periods of study and the institutions the strains were isolated from correlated significantly with the resistance patterns shown by the strains. The most frequent resistance patterns found were the following: ACC (6')-I at the Hospital Infantil de México (Children's Hospital of México), and ANT (2″) +AAC(6′)-I at the Instituto Nacional de Pediatría (INPed or National Institute of Pediatrics) in Mexico City. Furthermore, the isolation frequency of aminoglycoside-sensitive strains decreased remarkably at the INPed over a 12-year period. These results suggest that there has been a selection of Serratia marcescens strains that are very resistant to aminoglycosides.

Studies from developing countries suggest that persons with household-acquired (HA) measles are at greater risk of severe illness than persons with community acquired (CA) infection. Reported measles cases occurring among Milwaukee residents from May 1989 to June 1990 were used to assess whether household acquisition was a risk factor for severe measles in the United States. A case was classified as HA if onset of rash occurred 7–18 days after onset of rash in another case in the same household. Hospitalization rates were similar for 128 patients with HA measles (27%) and for 1004 patients with CA measles (26%). Multiple logistic regression was used to evaluate the association between hospitalization and household-acquisition after controlling for socioeconomic status, measles vaccination history, age, race, and date of onset of rash. Patients with HA measles were no more likely to be hospitalized than patients with CA measles (odds ratio 0·9, 95% confidence interval 0·6, 1·5). HA measles cases were not more severe than CA measles cases during this urban outbreak in the United States.