Provision and decontamination of uniforms in the NHS.

AIM: To ascertain the provision and decontamination of uniforms within a cross-section of NHS trusts in the UK and to compare policies regarding their use. METHOD: A questionnaire was circulated to 170 NHS trust infection control teams in the UK. Eighty-six (51 per cent) responses were received, which represented 101 NHS trusts. RESULTS: Less than half of the trusts (47 per cent) provide adequate numbers of uniforms to allow a clean uniform per shift. Only 26 per cent had adequate on-site staff changing facilities and 65 per cent did not launder uniforms. The majority of nursing staff (91 per cent) were compelled, by a combination of these factors, to launder their uniforms at home. Few were provided with any guidance on how to do this safely. CONCLUSION: There is an urgent need for minimum standards to be set for the provision of uniforms, laundering and changing facilities, to minimise the potential for spread of healthcare-associated infections.

An evaluation of the performance of XLD, DCA, MLCB, and ABC agars as direct plating media for the isolation of Salmonella enterica from faeces.

AIMS: To compare the performance of four media, singly and in combination, as direct plating media for the isolation of Salmonella enterica from human faeces. METHODS: Two thousand four hundred and nine routine, faecal samples received by four laboratories were inoculated on to xylose lysine desoxycholate (XLD), desoxycholate citrate (DCA), mannitol lysine crystal violet brilliant green (MLCB), and alpha-beta chromogenic (ABC) agars using standardised protocols, reagents, and data collection. Isolates of presumptive salmonellae were identified using standard laboratory techniques and the results were analysed statistically. RESULTS: Direct plating recovered 46 of the 60 possible isolates of Salmonella spp recovered via enrichment broth. No isolates were recovered from direct plating that were not recovered via selenite enrichment. MLCB gave the highest isolation rate individually (84.8%) and amounts of competing flora (CF) did not affect the recognition of colonies. ABC proved highly specific, but insensitive, and isolation rates were adversely affected by any amount of CF. Isolation rates from XLD and DCA were only affected when the CF load was heavy. DCA was least specific, with only 9.01% of picks positive and greatest number of confirmatory tests. XLD and MLCB, in combination, gave the highest isolation rate. CONCLUSIONS: Where the earlier results of direct plating may be advantageous, XLD and MLCB provide the optimal combination. For non-typhi salmonellae, MLCB is the best, single direct plating medium. For routine diagnostic work, XLD is most effective.

A comparison of the performance of cystine lactose electrolyte deficient (CLED) agar with Oxoid chromogenic urinary tract infection (CUTI) medium for the isolation and presumptive identification of organisms from urine.

AIMS: As part of the UK antimicrobial resistance strategy and action plan, the Public Health Laboratory Service (PHLS) is required to collect antibiotic susceptibility data so that resistance trends and patterns can be monitored. Most laboratories report urine Gram negative isolates, as "coliforms" according to morphological appearance, but without an acceptable identification system the antimicrobial surveillance data will be meaningless. Commercially available identification systems tend to be expensive and time consuming. Chromogenic agars, which claim to improve the detection of mixed cultures and identification of organisms from urine, have now become available and may provide a cost effective alternative. The primary aim of this study was to compare the performance of cystine lactose electrolyte deficient (CLED) agar with a chromogenic agar (Oxoid urinary tract infection medium; CUTI) in terms of isolation rates and ability to detect mixed cultures. Secondary aims were to evaluate the correlation of "presumptive" identification of isolates from chromogenic media with that of two commercial identification systems and to appraise the sensitivity of the semiquantitative loop and filter paper strip culture techniques. METHOD: One thousand, four hundred and sixty six urine samples were examined in four laboratories using the semiquantitative culture methods of 1 microl loop and filter paper strip. The degree of accuracy of organism identification was measured by comparing the presumptive identification using colony colour supplemented with simple bench tests, with identification obtained from two more complex commercial systems. RESULTS: There was no significant difference between the performance of the loop and filter paper strip methods on the CLED agar, but the CUTI agar performed significantly better than the CLED agar for the detection of significant isolates and mixed cultures. This difference was greater using the loop method. Identification of the organisms using the commercial systems gave > 99% agreement and was therefore considered suitable as a standard against which to compare the presumptive CUTI identification. Using the manufacturer's colony colour criteria in combination with a bench indole test, the CUTI medium was 99% specific for Escherichia coli, although this was reduced to 97% if the indole test was omitted. Citrobacter spp were the most commonly misidentified organisms, giving false presumptive identification as E coli. By testing oxidase activity to differentiate Pseudomonas spp and the absence of indole production to support the identification of Proteus mirabilis, the CUTI medium provided a suitable identification for 86.8% of Gram negative isolates. The remaining 13.2% would require further identification. CONCLUSION: CUTI medium improves the detection of mixed cultures, thereby improving the reliability of reporting of significant isolates when compared with CLED agar. When supplemented with simple bench tests it provides an identification system capable of speciating 86.8% of Gram negative isolates and providing a valuable cost effective mechanism for antimicrobial resistance surveillance.

A comparison of the performance of commercially available chromogenic agars for the isolation and presumptive identification of organisms from urine.

AIMS: To compare four media-UTI medium, BBL CHROMagar, CPS ID2, and Harlequin CLED-using a collection of fully characterised organisms and subsequent "field trial". METHODS: Seven hundred and eighty seven fully characterised isolates (730 Gram negative bacteria, 47 Gram positive bacteria, and 10 yeasts) were used to test for accuracy of organism identification. To assess isolation rates and ability to detect mixed cultures, 1435 urine samples were cultured in the three best performing chromogenic media (UTI medium, BBL CHROMagar, and CPS ID2) and CLED. RESULTS: The chromogenic agars differed in their accuracy of identification, with BBL CHROMagar performing best and Harlequin CLED performing least well. Similarly, BBL CHROMagar achieved a higher overall isolation rate than UTI medium and CPS ID2. When mixed growth was defined as greater than two organism types, BBL CHROMagar detected more mixed cultures than did UTI medium and CPS ID2, although the differences were not significant. When mixed growth was defined as greater than one organism type the increased number of mixed growths detected by BBL CHROMagar became significant, largely because of differences in enterococcal isolation rates. CONCLUSION: The use of BBL CHROMagar, UTI medium, or CPS ID2 chromogenic agar as a replacement for CLED agar would improve the detection rate of contaminated urine samples. Enhanced identification helps to distinguish different species, facilitating the monitoring of bacterial resistance in support of the national antibiotic strategy. BBL CHROMagar gave the highest overall organism recovery rates, greatest ability to detect mixed cultures, and the most accurate identification of organisms.

A comparison of blood agar supplemented with NAD with plain blood agar and chocolated blood agar in the isolation of Streptococcus pneumoniae and Haemophilus influenzae from sputum. Bacterial Methods Evaluation Group.

Streptococcus pneumoniae grows well and generally exhibits typical morphology on Columbia blood agar, whereas Haemophilus influenzae requires a more complex medium to meet its growth requirements - usually chocolated blood agar - on which S. pneumoniae is less easily recognisable. Therefore, a single medium that produces typical morphology of S. pneumoniae and facilitates the growth of H. influenzae would have considerable potential advantages. It has been claimed that blood agar supplemented with nicotinamide adenine dinucleotide (NAD) is such a medium. However, despite its routine use in several large diagnostic laboratories its performance has never been properly evaluated. In the present study, 1724 sputum samples were examined in four laboratories. The isolation rates of H. influenzae and S. pneumoniae on NAD-supplemented blood agar (SBA) were compared with those on a two-plate combination of plain blood (BA) and chocolated blood agar (CBA). The two-plate combination performed significantly better for both organisms; isolation rates for H. influenzae were increased from 8.16% on SBA to 11.07% on BA plus CBA and for S. pneumoniae from 4.18% to 4.68%. Isolation rates were also compared after incubation for 24 and 48 h. With the two-plate combination, isolation rates for H. influenzae and S. pneumoniae were increased by 0.98% and 0.16%, respectively, and for SBA by 0.57% and 0.32% after 48 h. However, despite this increase, SBA still performed less well than the two-plate combination.

A comparison of the isolation rates of Salmonella and thermophilic Campylobacter species after direct inoculation of media with a dilute faecal suspension and undiluted faecal material.

Regardless of media used, dilution of faecal samples before direct plating may improve isolation rates and reduce subcultures by freeing organisms from the faecal mass and diminishing competing flora. Despite the routine use of dilution in many laboratories, it has never been established properly whether direct or dilute inocula should be used in primary plating of faeces. A total of 3764 faecal samples was examined in four laboratories with a standardised methodology. The isolation rates, competing flora and confirmatory work performed for Salmonella spp. and Campylobacter spp. from primary plating media with a dilute faecal inoculum were compared with those after direct inoculation of faecal material. Inoculum effects on the isolation of Shigella spp. could not be assessed as only one isolate occurred during the study period. The overall isolation rates of both major enteric pathogens were unaffected by the inoculum. However, significantly fewer wasted subcultures were recorded with a dilute inoculum for Campylobacter spp., and competing florawas reduced in all cases without diluting out small numbers of the pathogen.

A comparison of the performance of bacitracin-incorporated chocolate blood agar with chocolate blood agar plus a bacitracin disk in the isolation of Haemophilus influenzae from sputum.

The lack of selectivity of chocolated blood agar (CBA), routinely used for the isolation of Haemophilus influenzae, may lead to masking of the growth of H. influenzae due to overgrowth of competing flora. Bacitracin can be used as a selective agent, either incorporated into the medium or applied to the medium in a filter paper. However, neither method has been evaluated or compared in a large study. Sputum samples (1990) were examined in four laboratories and the isolation rates of H. influenzae on chocolated blood agar with bacitracin added to the medium (BCA) and chocolated blood agar (CBA) with a bacitracin disk were compared. A plain blood agar plate was also inoculated to facilitate the isolation of Streptococcus pneumoniae so that its effects on the isolation of H. influenzae could be assessed. No significant difference was found between the isolation rates of H. influenzae on BCA and CBA with a bacitracin disk, although competing flora was greatly reduced and quantity of growth of H. influenzae increased on BCA. The presence of S. pneumoniae did not affect the isolation of H. influenzae in this study.

Antibiotic resistance and serotypes of Streptococcus pneumoniae at Birmingham Public Health Laboratory, 1989-94.

Antibiotic resistance of 1515 consecutive laboratory isolates of Streptococcus pneumoniae between 1989 and 1994 was analyzed. Overall, 39 (2.6%) isolates were resistant to penicillin, 102 (6.7%) resistant to erythromycin and 52 (3.4%) resistant to tetracycline. There was a higher proportion of penicillin resistant isolates from sterile sites compared with "non-sterile sites" (5% vs. 2.2%; P < 0.02). This same pattern occurred with erythromycin (12.5% vs. 5.6%; P < 0.001). From 1989-90 to 1993-94 the penicillin resistance rate increased from 0.8% to 8% and the erythromycin from 5.7% to 8.4%, whereas the tetracycline resistance rate fell from 3.7% to 2.8%. The increase in resistance to penicillin largely occurred in the final 12 months of this study period. One hundred and fifty isolates (9.9%) were serotyped, including isolates from sterile sites and those with penicillin resistance. The commonest serotypes of penicillin-sensitive pneumococci were 14, 19, 9 and 6. The majority of penicillin-resistant pneumococci (PRP) were of serotype 9 (64%) followed by 6, 23 and 19. Overall 95% of these isolates were of serotypes represented in the 23-valent pneumococcal polysaccharide vaccine (Pneumovax II). PRP were more likely to have resistance to erythromycin (23%) or tetracycline (23%) compared to penicillin-sensitive pneumococci (6% and 3% respectively). Most of the PRP were isolated from patients aged over 50 years including 11 isolates from blood cultures of patients with pneumonia or septicaemia. There was a possible epidemiological association between four patients with PRP but surveillance cultures of hospital contacts revealed no extra cases. These results show a worrying increase in infections due to PRP which has implications for clinical and laboratory staff in the diagnosis and treatment of serious pneumococcal infections.

Invasive candidiasis of the tonsil.

A patient with unilateral tonsillar enlargement secondary to locally invasive candidal infection is presented. This was not associated with any history of immunosuppression or prolonged use of broad-spectrum antibiotics. Invasive candidal infection of the tonsil has not previously been reported.

TGF-beta inhibition of CTL re-stimulation requires accessory cells and induces peroxisome-proliferator-activated receptor-gamma (PPAR-gamma).

Effective cellular immunity to Epstein-Barr virus (EBV), necessary to prevent or cure many post-transplant lymphoproliferative disorders (PTLD), can be inhibited by transforming growth factor-beta (TGF-beta). In vitro, TGF-beta inhibits memory CTL re-stimulation from whole PBMC. We show that the effect of TGF-beta on CTL re-stimulation is not directly on the T cell, but requires an accessory cell (AC) population. Further, pre-treatment of AC with TGF-beta significantly reduces memory CTL re-stimulation and suppresses delayed type hypersensitivity (DTH) responses. Addition of exogenous interferon-gamma to the AC overcomes the effects of TGF-beta. TGF-beta pre-treatment also up-regulates expression of peroxisome-proliferator-activated receptor-gamma (PPAR-gamma) in CD14(+) AC. Importantly, pre-treatment of AC with the PPAR-gamma ligand, ciglitazone, results in significantly reduced memory CTL re-stimulation. Thus, the effects of TGF-beta in this system may be mediated in part via PPAR-gamma, and PPAR-gamma activation could have significant inhibitory effects on memory T-cell responses by affecting AC function. These data have important implications in understanding how memory CTL are re-stimulated and function to prevent disease, especially PTLD.

An evaluation of immunomagnetic separation for the detection of salmonellas in raw chicken carcasses.

Immunomagnetic separation techniques were used in the isolation of salmonella from raw chicken carcasses. Improved isolation rates were achieved with increased specificity and decreased processing time, although several technical difficulties remain to be addressed. Immunomagnetic separation offers significant potential for improvement on existing microbiological systems for the isolation of salmonella.

A comparison of immunomagnetic separation plus enrichment with conventional Salmonella culture in the examination of raw sausages.

Immunomagnetic separation with additional enrichment was used in conjunction with improved selective media to improve the isolation of salmonellae from raw sausages. The isolation rate achieved was almost double that of conventional culture with no increase in processing time. The selective media gave an overall specificity of approximately 74%; all false-positive pickoffs being identified as Citrobacter freundii. It is believed that this method represents a significant advance in the isolation of salmonellae from foods, although the ideal media both for enrichment and selection have yet to be found.

Pharmacokinetics and tissue penetration of cefepime.

The pharmacokinetics of cefepime were studied following a 2 g intravenous infusion, by measuring concentrations in plasma, inflammatory fluid and urine. Mean peak plasma concentrations of 193.1 mg/l were achieved at the end of the 30 min infusion. The mean plasma elimination half-life was 2.1 h. Penetration into inflammatory fluid was rapid, with mean peak levels of 91.5 mg/l occurring 0.9 h after the end of the infusion. Urinary elimination accounted for 98.9% of the dose within 8 h. Therapeutic plasma levels (greater than 2 mg/l) were present for at least 8 h after the end of the infusion, suggesting that twice or three times daily dosing should be sufficient to treat infections due to susceptible organisms.

Comparison of the performance of selenite cystine and mannitol selenite enrichment broths in the isolation of Salmonella spp. from faeces.

Over a four-month period, 4,658 routine faecal samples were examined in four laboratories and the isolation rates of Salmonella spp. from mannitol selenite (MS) and selenite cystine (SC) broths plated to xylose lysine desoxycholate agar (XLD) compared. The isolation rate by MS was 1.55% and by SC was 1.48%, a small difference which is not statistically significant. Significantly fewer colonies were selected for supplementary testing from SC than MS (p = 0.029), thus reducing confirmatory work. In laboratories where SC is already used for food and environmental work, an opportunity exists to limit stocked salmonella enrichment broths to SC alone.

Effect of incubation temperature on the isolation of thermophilic species of Campylobacter from faeces.

Although no large-scale clinical study has been performed, it has been reported that incubation at 37 degrees C gives better isolation rates for all common species of Campylobacter than incubation at 42 degrees C, while also improving the recovery of the more unusual species. In this study, 2,570 faecal samples were examined in four laboratories, using a standardised protocol. Isolation rates of Campylobacter spp. were compared after incubation on modified campylobacter blood-free selective agar at 37 degrees C and 42 degrees C. Campylobacter spp. isolates were made from 185 samples (7.2%); 25 were recovered only at 42 degrees C and three only at 37 degrees C (p < 0.001). There were significantly more colonies at 42 degrees C (p = 0.001). Competing flora were heavier at 37 degrees C, but this did not account for the difference in isolation rates or quantity of growth. It is recommended that cultures for Campylobacter spp. be routinely incubated at 42 degrees C. More specific techniques are required to seek for fastidious campylobacters.

Comparison of the performance of lactose and mannitol selenite enriched broths, subcultured to DCA and XLD agars, in the isolation of Salmonella spp. from faeces.

Selenite-based enrichment broths using either lactose or mannitol as a carbohydrate source are generally used as selective enrichment media for the isolation of Salmonella spp. from human faeces in the UK, but few studies have compared the relative efficacy of the available formulations. A variety of solid media is used for the routine subculture from these selective broths, but similarly we have been unable to find published evidence as to which medium performs best. Four thousand and nineteen faecal samples were examined in four laboratories and the isolation rates of Salmonella spp. from lactose (LS) or mannitol selenite (MS) broths, plated onto either xylose lysine desoxycholate agar (XLD) or desoxycholate citrate agar (DCA) were compared. MS performed significantly better than LS (p = 0.02), recovering 95 salmonellae compared with 87. No significant difference in isolation rates was found between XLD and DCA, although colonial appearances of suspected salmonellae on XLD were much more specific, resulting in significantly fewer colonies having to be selected for supplementary testing (p < 0.001) and so reducing confirmatory work. An opportunity exists to simplify holdings of media by choosing to use the MS/XLD combination.