Campylobacteriosis outbreak associated with consumption of undercooked chicken liver pâté in the East of England, September 2011: identification of a dose-response risk.

A foodborne outbreak with 49 cases (22 culture positive for Campylobacter sp.) following a wedding party in the East of England was investigated. A retrospective cohort study identified an association between consumption of chicken liver pâté and infection with Campylobacter jejuni/coli. There was a statistically significant association between dose (amount of chicken liver pâté eaten) and the risk of disease ['tasted': odds ratio (OR) 1·5, 95% confidence interval (CI) 0·04-∞; 'partly eaten': OR 8·4, 95% CI 1·4-87·5; 'most or all eaten': OR 36·1, 95% CI 3·3-2119). The local authority found evidence that the preparation of chicken livers breached Food Standards Agency's guidelines. This epidemiological investigation established a clear dose-response relationship between consumption of chicken liver pâté and the risk of infection with Campylobacter. The continuing need to raise public awareness of the risk to human health posed by undercooked chicken liver is evident.

A recipe for disaster: outbreaks of campylobacteriosis associated with poultry liver pâté in England and Wales.

Despite the frequency of Campylobacter as the principal cause of bacterial gastroenteritis in the UK, outbreaks attributed to this pathogen are rare. One hundred and fourteen general foodborne outbreaks of campylobacteriosis were reported to the Health Protection Agency from 1992 to 2009 with most occurring in food service establishments (64%, 73/114). Poultry meat (38%, 43/114) was the most commonly reported vehicle of infection, of which poultry liver pâté, and undercooking, were strongly associated with this pathogen. Notably, the number of outbreaks of campylobacteriosis linked to consumption of poultry liver pâté in England and Wales increased significantly from 2007 (74% as opposed to 12%, P<0·00001) with a preponderance of these occurring in December. These outbreaks highlight the hazards associated with inappropriate culinary practices leading to undercooking of poultry liver pâté and suggest that improving catering practice is an important last line of defence in reducing exposure to Campylobacter-contaminated products.

Campylobacter and Salmonella in raw red meats in the United Kingdom: prevalence, characterization and antimicrobial resistance pattern, 2003-2005.

The prevalence of Campylobacter and Salmonella was assessed in 3959 raw red meats in the UK during 2003-2005. Meats were more frequently contaminated with Campylobacter (7.2%) than with Salmonella (2.4%). Lamb and other meats (e.g. mutton, rabbit) exhibited the highest contamination from Campylobacter (12.6% and 19.8%, respectively), compared with pork (6.3%) and beef (4.9%). Pork however had the highest contamination from Salmonella (3.9%), followed by lamb (2.0%), other meats (2.0%) and beef (1.3%). Offal samples (36.6%) were more frequently contaminated with Campylobacter or Salmonella than muscle tissue (7.0%). C. jejuni predominated in all meat types. C. coli isolates were more likely to exhibit antimicrobial drug resistance, including quinolones, than C. jejuni. Salmonella typhimurium was the most frequent Salmonella serotype isolated from meats; S. typhimurium DT104/104b isolates exhibited higher rates of multiple drug resistance than other serotypes. The findings reinforce the importance of adequate cooking of meat and good hygiene to avoid cross-contamination.

Cyheptamide and 3-hydroxy-3-phenacyloxindole: structural similarity to diphenylhydantoin as the basis for anticonvulsant activity.

The molecular structures of cyheptamide and 3-hydroxy-3- phenacyloxindole were determined by X-ray diffraction methods. The amide group in both compounds exhibits delocalization of the pi-electrons over the three atoms (N, C, and O), while the bond linking the amide to the tetrahedral carbon atom is a single bond. These structural features are also present in two drugs used for the treatment of generalized tonic-clonic ( GTC ) seizures, namely, carbamazepine and diphenylhydantoin. The shapes of cyheptamide , 3-hydroxy-3- phenacyloxindole , and carbamazepine have three features that are the same and can be simultaneously overlapped, the amide and two hydrophobic regions, whereas diphenylhydantoin fits two of the three regions at one time. These structural and electronic features are analyzed in light of current models for anticonvulsant activity.

The induction of reciprocal translocations in rhesus monkey stem-cell spermatogonia: effects of low doses and low dose rates.

The induction of reciprocal translocation in rhesus monkey spermatogonial stem cells was studied following exposure to low doses of acute X rays (0.25 Gy, 300 mGy/min) or to low-dose-rate X rays (1 Gy, 2 mGy/min) and gamma rays (1 Gy, 0.2 mGy/min). The results obtained at 0.25 Gy of X rays fitted exactly the linear extrapolation down from the 0.5 and 1.0 Gy points obtained earlier. Extension of X-ray exposure reduced the yield of translocations similar to that in the mouse by about 50%. The reduction to 40% of translocation rate after chronic gamma exposure was clearly less than the value of about 80% reported for the mouse over the same range of dose rates. Differential cell killing with ensuing differential elimination of aberration-carrying cells is the most likely explanation for the differences between mouse and monkey.

Evaluation of a micromethod gallery (API Staph) for the identification of staphylococci and micrococci.

A collection of 300 well-characterised strains of staphylococci and micrococci was examined by a commercially available gallery micromethod (API Staph). The results were compared with biotyping by conventional methods. The gallery micromethod broadly agreed with the biotyping scheme used but gave an identification from the index supplied in less than 30% of the trials. Reproducibility was better after 48 h incubation than after 24 h but was poor for the tests for phosphatase and acetoin. When compared with the results of conventional tests, the tests for acetoin, phosphatase and urea were unsatisfactory.

Changing resistance to antimicrobial drugs, and resistance typing in clinically significant strains of Staphylococcus epidermidis.

Resistance to 11 antimicrobial drugs was assessed in 532 clinically significant strains of Staphylococcus epidermidis received in the years 1978, 1979 and 1980 and compared with that of strains collected in 1976 and 1977 for an international study. Only 14% of strains were sensitive to all the drugs tested. Resistance to gentamicin increased significantly from 7% to 33% during the study period. The degree of association between pairs of drugs was assessed. There was strong association between resistance to methicillin, aminoglycosides and macrolide antibiotics and only weak association between resistance to novobiocin, tetracycline, chloramphenicol and penicillin. Patterns of resistance were complex and may be useful as an accessory typing system.

Micrococcus in the blood.

Eight isolates of micrococci from the bloodstream of six patients obtained under circumstances suggesting a pathogenic role were studied in detail. The organisms were remarkably uniform in cultural, biochemical and antibiotic-susceptibility characters. All strains showed high resistance to methicillin and hydrolysed arginine. The characters found did not correspond with those of any hitherto described species, but were closest to Micrococcus lylae.

Supplementary phages for the investigation of strains of methicillin-resistant Staphylococcus aureus.

Nineteen experimental phages were derived by mitomycin-C induction from methicillin-resistant strains of Staphylococcus aureus collected world-wide. They were assessed for their ability to distinguish isolates of a methicillin-resistant strain of S. aureus epidemic in the London area from other British strains, both sensitive and resistant to methicillin. The experimental phages were most active against strains of phage groups III and I + III. One phage was related to the phages of lytic group I. A typing pattern common to isolates of the epidemic strain was identified and used as an aid in the recognition of this strain. Ten of the phages were retained for further study.

Epidemiological typing of coagulase-negative staphylococci from nosocomial infections.

Biotyping, antibiograms, bacteriophage typing, plasmid profile analysis and SDS-PAGE protein profiles were used to determine the relatedness of 44 Staphylococcus epidermidis and four S. haemolyticus isolates from 14 patients. A selection of these were further characterised by ribotyping. Biotyping classified the isolates into three major groups but was considered a poor strain marker. Although antibiograms classified the S. epidermidis isolates into 20 groups, some changes in the susceptibility patterns of related isolates from a single patient were demonstrated. Bacteriophage typing was the least discriminatory of the methods used. SDS-PAGE gave highly related patterns for the majority of S. epidermidis isolates. Plasmid profile analysis and ribotyping, with a minimum of two restriction endonucleases, were the most discriminatory methods for typing S. epidermidis. Nonetheless, some isolates from the same patient - probably representing a single strain - varied in plasmid profile indicating plasmid instability. One of six related isolates from a single patient lacked two bands from the ribotyping pattern of the other isolates. Although no single method proved entirely satisfactory on all occasions, the combination of typing methods was sufficient to provide evidence of the relatedness of S. epidermidis isolates from individual patients.

Evaluation of methods for typing coagulase-negative staphylococci.

One hundred and forty-two coagulase-negative staphylococci (CNS) isolated from dialysate effluent or skin of patients receiving continuous ambulatory peritoneal dialysis (CAPD) were typed by extended antibiogram (16 antibiotics) and biotype (26 reactions). These isolates were then typed by supplementary methods to determine the most suitable typing method for an epidemiological study of antibiotic resistance. These included phage typing, reverse phage typing, plasmid typing, whole-cell protein typing by SDS-PAGE with analysis by densitometry, and immunoblotting. The percentage of isolates typed successfully by the supplementary methods were: phage typing 20%, reverse phage typing 0%, plasmid typing 66%, SDS-PAGE 100%, immunoblotting 100%. The discrimination of each method was: phage typing 20%, plasmid typing 37%, SDS-PAGE 69%, immunoblotting 57%. Reproducibility was 88% for phage typing and 97% for plasmid typing. The reproducibility of the whole-cell protein typing was 83% if the same extracts were used but only 43% when separate protein extracts were analysed on separate occasions. However, strain relatedness was highly reproducible. The determination of an antibiogram-biotype profile was not a sufficiently accurate typing method for an epidemiological study of antibiotic resistance. Whole-cell protein typing by SDS-PAGE or immunoblotting was technically demanding but was the most effective of the supplementary methods for detecting erroneous discrimination and false matching produced by antibiogram-biotype combinations.

Evaluation of electrophoretic methods for typing methicillin-resistant Staphylococcus aureus.

Three electrophoretic methods of typing methicillin-resistant Staphylococcus aureus (MRSA) strains--plasmid profiles (PP), whole-cell protein profiles (WCPP) and immunoblotting profiles (IP)--were evaluated and compared with phage typing. The results obtained with isolates from 12 outbreaks were compared both within the outbreaks, to determine the consistency of results, and between outbreaks. There was generally good agreement between the typing methods but in only six outbreaks did all four methods indicate the same relationship between isolates. WCPP comprised more than 50 bands; when differences occurred, they were seen in only a few bands. In contrast, IP comprised only one or two major bands and the differences were much easier to interpret. The PPs of many of the isolates were similar; many isolates contained a plasmid of mol. wt (18-25) x 10(6). In several outbreaks both WCPP and IP showed minor differences between isolates that were not apparent with phage typing. When comparisons were made between the 12 index strains and an isolate representing the London epidemic MRSA strain, phage typing and WCPP were the most discriminatory methods; both gave nine distinct patterns, whereas there were eight IPs and only six PPs amongst the 13 strains. It was concluded that both WCPP and IP could provide valuable epidemiological data on MRSA and that IP was the easiest of the three methods to interpret.

Characterization of a strain of methicillin-resistant Staphylococcus aureus (EMRSA-15) by conventional and molecular methods.

A strain of methicillin-resistant Staphylococcus aureus (MRSA), EMRSA-15, was isolated in both the Midlands and south-east of England. This strain could be distinguished from another, very similar strain, found in the north of England, by both conventional and molecular typing. Conventional typing allowed distinction between the Midlands and southern variants of EMRSA-15, while molecular typing (pulse-field gel electrophoresis) allowed recognition of local variants in the south. In this investigation conventional and molecular typing methods were complementary.

Characters of coagulase-negative staphylococci and micrococci from cases of endocarditis.

Strains of coagulase-negative staphylococci and micrococci submitted to a reference laboratory from 137 cases of endocarditis over a 5 year period were reviewed. Staphylococcus epidermidis biotype 1 (SII) was the commonest biotype in all groups of patients but exceeded 80 per cent in the 61 patients who had undergone prosthetic valve surgery and the 16 patients who had undergone other forms of surgery. Biotype 4 (SVI) was recovered from 10 of the 34 patients without surgery. Strains from prosthetic valve endocarditis were frequently resistant to many antibiotics while strains from natural valve endocarditis were frequently sensitive to all, or resistant only to penicillin. The value of bacteriophage typing was confirmed.

PCR-RFLP analysis of the coagulase gene of Staphylococcus aureus: application to the differentiation of epidemic and sporadic methicillin-resistant strains.

Preventing cross-infection with epidemic strains of methicillin-resistant Staphylococcus aureus (MRSA) requires effective control measures. These call for simple, rapid, discriminatory and reproducible methods for typing this pathogen. In this study 140 isolates/strains from 105 hospitals in England and Wales, representing 72 diverse phage types, were analysed by bacteriophage typing and PCR coagulase (coa) gene restriction fragment length polymorphism (RFLP). Isolates gave a coa gene PCR product that was either 660 base pairs (bp), 603 bp or 547 pb in size. The PCR products were digested with Alu I and Cfo I, and the fragments separated by gel electrophoresis. Eight coa gene RFLP patterns, numbered 1 to 8, were observed. Pattern 3 was most common (N = 25 isolates), followed by patterns 2 and 5 (18 isolates each), pattern 1 (14 isolates), pattern 4 (11 isolates), pattern 7 (10 isolates), pattern 8 (eight isolates) and pattern 6 (six isolates). Isolates of the same phage type often gave different coa gene RFLP patterns, and the patterns within the epidemic types EMRSA-03, EMRSA-15 and EMRSA-16 were heterogeneous. Thus, representatives of EMRSA-03 were subtyped to coa RFLP patterns 1 and 2, those of EMRSA-05 to coa RFLP patterns 1, 2, 7 and 8, and those for EMRSA-16 to coa RFLP patterns 2, 3, 4, 5 and 6. The range of patterns within single phage types of S. aureus could help to discriminate between isolates/strains, and in a hierarchical approach coa gene RFLP could occupy an intermediate position between phage typing and pulsed-field gel electrophoresis (PFGE).

Beta-lactamase-negative, methicillin-resistant Staphylococcus aureus in a newborn nursery: report of an outbreak and laboratory investigations.

An outbreak of skin infection caused by a beta-lactamase-negative strain of methicillin-resistant Staphylococcus aureus (MRSA) occurred during a five-week period in a newborn nursery. Twelve babies, two mothers and two members of staff were involved. One baby had a diagnosis of staphylococcal scalded skin syndrome and two others required treatment with antibiotics. The infecting strain produced exfoliative toxin A. It was thought that it had been introduced from a different maternity unit by a nasal carrier. Laboratory investigations tended to support this hypothesis.

Adhesive tapes in the special care baby unit.

Semi-quantitative sampling of 37 taped sites and 37 matching control sites in 30 babies in a special care baby unit showed a tendency for bacterial growth under the cardiac monitors or the occlusive plastic adhesive tape to be greater than at the control site. This tendency was demonstrable even when potential pathogens were considered. During the study, an outbreak due to Candida albicans occurred. Antibiotic resistance of coagulase-negative staphylococci was marked and this, and other evidence, suggested that part of the skin flora was acquired from the unit.

Use of fluorescent amplified fragment length polymorphism (fAELP) to characterise methicillin-resistant Staphylococcus aureus.

The new PCR-based genotyping technique, fluorescent amplified fragment length polymorphism (fAFLP), was compared for discriminatory power and reproducibility with standard phenotypic methods, a coagulase gene (coa) restriction fragment length polymorphism (RFLP) method and pulsed-field gel electrophoresis (PFGE), in typing 34 isolates and four reference strains of methicillin-resistant Staphylococcus aureus (MRSA). The fAFLP showed from 40 to 75 fragments, 50 to 450 base pairs (bp) in size. Based on replicate studies, the isolates were judged indistinguishable when their fAFLP pattern was >93.7% similar. Only two of the isolates were indistinguishable by this criterion. Thirty-one MRSA fell into four major fAFLP groups (1, 2, 3 and 4) at the level of >79.9% similarity. Three other isolates and an EMRSA-16 strain fell outside these major groups. Within both fAFLP groups 1 and 2, two subgroups, A and B, could be identified at approximately 82.0% similarity. While most isolates within group 1 could also be separated by their phenotypic and coagulase gene (coa) RFLP pattern, all the isolates within fAFLP groups 2A and 2B were identical on the basis of these characters. The MRSA within fAFLP groups 3 and 4 were heterogeneous by their phenotypic characteristics and coa gene RFLP patterns. fAFLP was reproducible and distinguished between MRSA isolates that appeared identical by other methods. It is likely to contribute to the epidemiological analysis of outbreaks of MRSA infection.