Validation of an antimicrobial susceptibility testing protocol for Brachyspira hyodysenteriae and Brachyspira pilosicoli in an international ring trial.

Brachyspira hyodysenteriae and Brachyspira pilosicoli cause economically important enteric disease in pigs. Treatment of these infections often includes antimicrobial administration, which can be most effective when therapeutic options are informed by antimicrobial susceptibility testing data. Here we describe a method for broth dilution antimicrobial susceptibility testing of these bacteria, both of which are difficult to culture in vitro. The protocol was evaluated for its fitness for use in an inter-laboratory ring trial involving eight laboratories from seven countries, and employing eleven test strains (5 Brachyspira hyodysenteriae including the type strain B78T and 6 Brachyspira pilosicoli) and six antibiotics. Overall intra- and inter-laboratory reproducibility of this method was very good (>90 % MICs at mode +/- 1 log2). Whole genome sequencing revealed good correspondence between reduced susceptibility and the presence of previously defined antimicrobial resistance determinants. Interestingly, lnu(C) was identified in B. pilosicoli isolates with elevated MICs of lincomycin, whilst tva(B) was associated with elevated MICs of pleuromutilins in this species. We designated two new control strains with MICs lying within currently tested ranges, including for the pleuromutilins, in contrast to the control strain B. hyodysenteriae B78T. These were deposited at the DSMZ-German Collection of Microorganisms and Cell Cultures GmbH. The validation of a standard protocol and identification of new control strains facilitates comparisons between studies, establishment of robust interpretative criteria, and ultimately contributes to rational antimicrobial use when treating infected livestock.

A five-year follow-up observational study of the T-14 paediatric throat disorders outcome measure in tonsillectomy and adenotonsillectomy.

INTRODUCTION: The T-14 questionnaire is a validated outcome measurement tool to assess the value of paediatric tonsillectomy from a parent's perspective. There is a paucity of data regarding the long-term postoperative effects of tonsillectomy on quality of life in the paediatric population. Our previous study assessed T-14 scores up to year 2 postoperatively, with this study extending follow-up to 5 years. MATERIALS AND METHODS: We undertook a prospective uncontrolled observational study examining 54 paediatric patients undergoing tonsillectomy at Portsmouth Hospitals NHS Trust. Parents of children undergoing surgery were invited to complete a T-14 questionnaire preoperatively, as well as at 3 months, 6 months, 1 year, 2 years and now 5 years postoperatively. RESULTS: In total, 44 of 54 patients completed questionnaires preoperatively and at all postoperative time points, with 46 being completed at 5 years. There was a highly significant (P < 0.001) difference between the preoperative scores and all other measured T-14 scores postoperatively. The mean score preoperatively was 33.3 compared with 1.0 at 5 years. CONCLUSIONS: This is the first study to assess long-term quality of life following paediatric tonsillectomy using the T-14 questionnaire. The benefits of tonsillectomy on long-term quality of life further confirms its value within the paediatric population.

Antimicrobial use and antimicrobial susceptibility in Escherichia coli on small- and medium-scale pig farms in north-eastern Thailand.

BACKGROUND: Intensification of livestock production seen in many low- and middle-income countries is often believed to be associated with increased use of antimicrobials, and may hence contribute to the emergence of antimicrobial resistance. The aim of this study was to map antimicrobial use on small- (n = 25) and medium-scale (n = 27) pig farms in north-eastern Thailand, and to compare antimicrobial susceptibility of commensal Escherichia coli isolated from sows on these farms. METHODS: Information regarding pig husbandry and antimicrobial treatment regimens was obtained by the use of semi-structured questionnaires. Faecal samples were collected from three healthy sows at each farm, and Escherichia coli was cultured and analysed for antimicrobial susceptibility using the broth microdilution method. Multilevel regression models were used to compare antimicrobial susceptibility between isolates from small- and medium-scale farms. RESULTS: All farms included in the study administered antimicrobials to their sows. Small-scale farmers most commonly (64%) decided themselves when to give antimicrobials and the majority (60%) bought the medicines at the local store or pharmacy, whereas farmers on medium-scale farms always discussed antimicrobial treatment with a veterinarian. Medium-scale farms used a greater diversity of antimicrobials than small-scale farms and did also administer antimicrobials in feed to a higher extent. High levels of antimicrobial resistance to several critically important antimicrobials for human medicine (including ciprofloxacin, streptomycin and ampicillin) were found in isolates from both small- and medium-scale farms. Resistance levels were significantly (P < 0.05) higher in isolates from medium-scale farms for several of the antimicrobials tested, as well as the level of multidrug-resistance (P = 0.026). CONCLUSION: The routines regarding access and administration of antimicrobials differed between the small- and medium-scale farms. Although the level of antimicrobial resistance, as well as multidrug-resistance, was higher in isolates from medium-scale farms, it cannot be concluded if this increase is a consequence of a more abundant use of antimicrobials, or a result of differences in administration routines.

A two-year follow-up observational study of the T-14 paediatric throat disorders outcome measure in tonsillectomy and adenotonsillectomy.

INTRODUCTION: The T-14 questionnaire is a validated patient reported outcome measure used to assess the value of paediatric tonsillectomy from the patient's perspective. There are currently limited data revealing the long-term postoperative effects of tonsillectomy on quality of life in the paediatric population. A previously published study was therefore extended to provide additional data at 12 and 24 months following surgery. METHODS: A prospective uncontrolled observational study was undertaken examining 54 paediatric patients undergoing tonsillectomy at Portsmouth Hospitals NHS Trust. Parents of children having surgery were invited to complete a T-14 questionnaire preoperatively as well as at 3, 6 (previously published), 12 and 24 months postoperatively. RESULTS: The questionnaire was completed for 50 of the 54 patients preoperatively as well as at 3, 6 and 12 months postoperatively, with 44 being completed at 24 months. The mean difference between the preoperative T-14 scores and the scores at 3, 6, 12 and 24 months following surgery were highly statistically significant (p<0.001). CONCLUSIONS: This is the first study published in the literature to assess the T-14 questionnaire at 12 and 24 months following paediatric tonsillectomy, providing evidence of the ongoing benefit of patient reported outcome measures. This further confirms the value of tonsillectomy in the paediatric population and demonstrates its ongoing positive effects on quality of life.

Patient information leaflet on mastoid surgery risks: assessment of readability and patient understanding.

INTRODUCTION: It is important that patients have a good understanding of surgery-related risks, particularly for mastoid surgery, which exposes patients to the risk of very serious complications, despite addressing conditions which often have only minor symptoms. MATERIALS AND METHODS: A patient information leaflet describing the risks of mastoid surgery was prepared. However, the Hospital Patient Advice and Liaison Services team thought it was too long and complicated. It was introduced unchanged. Fifty-four consecutive mastoidectomy patients were given a questionnaire asking for their opinion of the leaflet. The leaflet was also assessed with readability formulae and the Ensuring Quality Information for Patients tool. RESULTS AND ANALYSIS: Ninety-eight per cent of respondents thought the leaflet's writing style was easy to understand. The majority (96 per cent) thought the length was 'just right'. The 7 readability formulae used established readability at a grade 9 level (i.e. appropriate for a reading age of 13-15 years). The Ensuring Quality Information for Patients score was 87.5 per cent. DISCUSSION: Despite the drive to simplify patient information leaflets, quite detailed information is sometimes required. A style which is too simple may be perceived as patronising and may encourage patients to underestimate potential risks. It is important to ask patients their opinion.

Appropriate maximum holding times for analysis of total suspended solids concentration in water samples taken from open-channel waterways.

Many scientific laboratories follow, as standard practice, a relatively short maximum holding time (within 7 days) for the analysis of total suspended solids (TSS) in environmental water samples. In this study we have subsampled from bulk water samples stored at ∼4 °C in the dark, then analysed for TSS at time intervals up to 105 days after collection. The nonsignificant differences in TSS results observed over time demonstrates that storage at ∼4 °C in the dark is an effective method of preserving samples for TSS analysis, far past the 7-day standard practice. Extending the maximum holding time will ease the pressure on sample collectors and laboratory staff who until now have had to determine TSS within an impractically short period.

Cutaneous sensory deficit following post-auricular incision.

INTRODUCTION: Post-auricular incisions are performed for a range of otological procedures. Anecdotally, many patients suffer some numbness of the pinna or post-auricular skin post-operatively, but for most this appears to reduce with time. This study aimed to investigate this phenomenon. METHODS: A single centre, questionnaire-based study was undertaken, assessing the presence of numbness beyond eight months post-surgery, its location, how it changed, and its impact on the patient. Patients whose numbness had resolved provided details of any temporary deficit. Data were acquired for 35 primary and 16 revision procedures. RESULTS AND CONCLUSION: Sixty-nine per cent of patients undergoing primary surgery experienced post-operative numbness. Twenty-six per cent had continued numbness after at least eight months' recovery, but only 3 per cent were constantly aware of the deficit. Of those with an ongoing deficit, 78 per cent felt there had been a reduction in the severity, and 67 per cent in the area size, over time. Recovery appeared to be slightly worse in revision cases.

TSC-22D1 isoforms have opposing roles in mammary epithelial cell survival.

Transforming growth factor beta (TGFbeta)-stimulated clone-22 domain family member 1 (TSC-22D1) has previously been associated with enhanced apoptosis in several cell systems. In an attempt to identify novel factors that are involved in the control of cell death during mammary gland involution, we found that the mRNA for isoform 2 of TSC-22D1 was highly upregulated 24 h after forced weaning, when a dramatic increase in cell death occurred, closely following the expression of the known inducer of cell death during involution, TGFbeta3. This was paralleled by strongly increased TSC-22D1 isoform 2 protein levels in the luminal epithelium. In contrast, RNA and protein expression levels of the isoform 1 of TSC-22D1 did not change during development. Whereas isoform 2 induced cell death, isoform 1 suppressed TGFbeta-induced cell death and enhanced proliferation in mammary epithelial cell lines. Furthermore, four distinct forms of isoform 2 protein were detected in the mammary gland, of which only a 15-kDa form was associated with early involution. Our data describe novel opposing functions of the two mammalian TSC-22D1 isoforms in cell survival and proliferation, and establish the TSC-22D1 isoform 2 as a potential regulator of cell death during mammary gland involution.

Characterization and epidemiological relationships of Spanish Brachyspira hyodysenteriae field isolates.

This research aimed to describe the genetic and phenotypic diversity of 74 Spanish Brachyspira hyodysenteriae field isolates, to establish epidemiological relationships between the isolates and to confirm the presence of tiamulin-resistant isolates in Spain. For these purposes, we performed biochemical tests in combination with diagnostic PCR analysis for the identification of Brachyspira spp. and for detection of the smpA/smpB gene. We also used antimicrobial susceptibility tests, random amplified polymorphic DNA (RAPD) and a new pulsed-field gel electrophoresis (PFGE) protocol. The combination of RAPD and PFGE allowed the study of epidemiological relationships. Both indole-negative and tiamulin-resistant isolates of B. hyodysenteriae are reported in Spain for the first time. The genetic analyses indicated a relationship between these Spanish isolates and indole-negative isolates previously obtained from Germany and Belgium.

Uptake of pneumococcal polysaccharide vaccine in at-risk populations in England and Wales 1999-2005.

The UK has had a pneumococcal polysaccharide vaccination (PPV) programme for groups at higher risk of invasive disease since 1992. This paper presents data from a sample of primary-care practices (Q-RESEARCH) of PPV uptake in patients according to their risk status. Of 2.9 million registered patients in 2005, 2.1% were vaccinated with PPV in the preceding 12 months and 6.5% in the preceding 5 years. Twenty-nine per cent of the registered population fell into one or more risk groups. The proportion of each risk group vaccinated in the previous 5 years ranged from 69% (cochlear implants), 53.4% (splenic dysfunction), 36.5% (chronic heart disease), 34.7% (diabetes), 22.9% (immunosuppressed), 28.7% (chronic renal disease), 15.9% (sickle cell disease) to 12.6% (chronic respiratory disease). Uptake was lower in areas where the non-white proportion of population was >10%. In conclusion, there remain large gaps in the uptake of PPV in several high-risk populations in the United Kingdom. Effective strategies need to be developed to address these deficiencies.

The Picture Archiving and Communication System: a useful tool in a case of upper oesophageal foreign body.

We present a case of an oesophageal foreign body (approximately 2x2 cm) which was not seen on the initial lateral soft tissue neck X-ray but was detected following manipulation of the X-ray using the Picture Archiving and Communication System of X-ray viewing. This system gives the user much greater flexibility, including a choice of magnification and contrast.

Quality-control ranges for antimicrobial susceptibility testing by broth dilution of the Brachyspira hyodysenteriae type strain (ATCC 27164T).

There are no approved standards for antimicrobial susceptibility testing of the fastidious spirochete Brachyspira hyodysenteriae. An interlaboratory study was performed to establish MIC quality control ranges for six antimicrobial agents for the type strain of B. hyodysenteriae using broth dilution. The results showed that B. hyodysenteriae B78T ATCC 27164T is a suitable quality control strain. This is a first step toward standardization of methods regarding this anaerobe.

Self-reported benefits from successive bilateral cochlear implantation in post-lingually deafened adults: randomised controlled trial.

Adult users of unilateral Nucleus CI24 cochlear implants with the SPEAK processing strategy were randomised either to receive a second identical implant in the contralateral ear immediately, or to wait 12 months while they acted as controls for late-emerging benefits of the first implant. Twenty four subjects, twelve from each group, completed the study. Receipt of a second implant led to improvements in self-reported abilities in spatial hearing, quality of hearing, and hearing for speech, but to generally non-significant changes in measures of quality of life. Multivariate analyses showed that positive changes in quality of life were associated with improvements in hearing, but were offset by negative changes associated with worsening tinnitus. Even in a best-case scenario, in which no worsening of tinnitus was assumed to occur, the gain in quality of life was too small to achieve an acceptable cost-effectiveness ratio. The most promising strategies for improving the cost-effectiveness of bilateral implantation are to increase effectiveness through enhanced signal processing in binaural processors, and to reduce the cost of implant hardware.

Clinical pathways for patients with newly diagnosed hepatitis C - what actually happens.

Management of hepatitis C virus (HCV)-infected individuals requires referral to specialist care. To determine whether patients newly diagnosed as anti-HCV positive are appropriately referred for further investigation and management, and if not, to determine why not. We studied patients tested for antibodies to HCV by Nottingham Public Health Laboratory in a 2-year period (2000-2002). The progress of newly diagnosed anti-HCV positive patients into specialist clinics for further management was documented. For patients not referred for specialist care, a questionnaire was sent to the clinician requesting the initial anti-HCV test, to identify reasons for nonreferral. Eleven thousand one hundred and seventy-seven patients were tested for anti-HCV. Two hundred and fifty-six (2.3%) were newly diagnosed as being anti-HCV positive. Two per cent of samples sent from primary care were anti-HCV positive, compared to 18.8, 18.9 and 1.3% sent from prison, drug and alcohol units, and secondary care, respectively. About 64.3% of positive patients diagnosed in primary care were referred to specialist care, compared to 18.4, 42.4 and 62.6% of patients diagnosed in the other three settings. One hundred and twenty-five (49%) newly diagnosed patients were referred appropriately for further management. 68 of these attended clinic, 45 underwent liver biopsy and 26 (10%) began treatment. One hundred and thirty-one patients (51%) were not referred. In 54 cases, there was no evidence that the anti-HCV positive result reached the patient. In 15, referral was considered but rejected, and 20 patients were referred to non-HCV-specialists (their general practitioners or to genito-urinary medicine). Hence less than 50% of newly diagnosed anti-HCV positive patients are referred to an appropriate clinic for further investigation and management. Reasons for this are multifarious and complex, reflecting both systems failure and patient choice. Unless these are understood and addressed, the Department of Health Hepatitis C Strategy (2002) and Action Plan for England (2004) will fail to achieve their intended objectives.

Tiamulin resistance in porcine Brachyspira pilosicoli isolates.

There are few studies on antimicrobial susceptibility of Brachyspira pilosicoli, therefore this study was performed to investigate the situation among isolates from pigs. The tiamulin and tylosin susceptibility was determined by broth dilution for 93 and 86 porcine B. pilosicoli isolates, respectively. The isolates came from clinical samples taken in Swedish pig herds during the years 2002 and 2003. The tylosin minimal inhibitory concentration (MIC) was >16 microg/ml for 50% (n=43) of the isolates tested. A tiamulin MIC >2 microg/ml was obtained for 14% (n=13) of the isolates and these were also tested against doxycycline, salinomycin, valnemulin, lincomycin and aivlosin. For these isolates the susceptibility to salinomycin and doxycycline was high but the MICs for aivlosin varied. The relationship between the 13 tiamulin resistant isolates was analyzed by pulsed-field gel electrophoresis (PFGE). Among the 13 isolates 10 different PFGE patterns were identified.

Assessment of diagnostics and antimicrobial susceptibility testing of Brachyspira species using a ring test.

There is no ring test for quality assessment available in Europe for diagnostics and antimicrobial susceptibility testing of the fastidious, anaerobic bacteria of the genus Brachyspira. Therefore, an international ring test for Brachyspira spp. was performed once a year during 2002-2004. Two sets of coded samples were prepared and distributed on each occasion. One set comprised six swabs dipped in pig faeces spiked with Brachyspira spp. intended for diagnostics. The other set comprised two pure strains intended only for susceptibility testing. All methods used were in-house methods. The species used were Brachyspira hyodysenteriae, Brachyspira pilosicoli, Brachyspira innocens, Brachyspira murdochii and Brachyspira intermedia. In most cases, the correct Brachyspira spp. were detected. However, the results showed that Brachyspira spp. could be difficult to identify, especially if two Brachyspira spp. were mixed or if the concentration of Brachyspira in faeces was low. Additionally, some laboratories reported Brachyspira growth in control samples that were not seeded with any spirochaetes. The lowest detection level was 10(2) bacteria/ml faeces for both B. hyodysenteriae and B. pilosicoli. The susceptibility tests performed showed that disc diffusion was not recommendable for Brachyspira spp. Extended antimicrobial dilution series gave most congruent results. The diversity of the results highlights the importance of ring tests for a high quality of diagnostics and antimicrobial susceptibility tests for Brachyspira spp. This is the first ring test described for Brachyspira spp.

Radiological and radionuclide investigation of malignant otitis externa.

This study looks at case series of malignant otitis externa, outlines detailed structural (radiological) and functional (radionuclide) investigations, and discusses their utility in the initial diagnosis, patient management and follow up of this condition. Patients were investigated by computerized tomography (CT), magnetic resonance imaging (MRI), two-phase planar and single-photon emission tomography (SPECT), technetium-99m methylenediphosphonate bone scans, gallium-67 planar and SPECT scans, and indium-111 or technetium-99m labelled WBC planar and SPECT scintigraphy. We discuss example case reports with the various radiological and scintigraphic findings and outline a protocol for rational investigation. It is concluded that CT and/or MRI should be supported by routine SPECT bone imaging for initial diagnosis of malignant otitis externa. Routine SPECT bone imaging further supplemented by gallium scintigraphy should be the investigation of choice in the follow up of these cases for assessing response to treatment and disease recurrence.

Testis-specific TAF homologs collaborate to control a tissue-specific transcription program.

Alternate forms of the PolII transcription initiation machinery have been proposed to play a role in selective activation of cell-type-specific gene expression programs during cellular differentiation. The cannonball (can) gene of Drosophila encodes a homolog of a TBP-associated factor (dTAF5) protein expressed only in spermatocytes, where it is required for normal transcription of genes required for spermatid differentiation. We show that Drosophila primary spermatocytes also express four additional tissue-specific TAFs: nht (homolog of dTAF4), mia (homolog of dTAF6), sa (homolog of dTAF8) and rye (homolog of dTAF12). Mutations in nht, mia and sa have similar effects in primary spermatocytes on transcription of several target genes involved in spermatid differentiation, and cause the same phenotypes as mutations in can, blocking both meiotic cell cycle progression and spermatid differentiation. The nht, mia, sa and rye proteins contain histone fold domain dimerization motifs. The nht and rye proteins interact structurally when co-expressed in bacteria, similarly to their generally expressed homologs TAF4 and TAF12, which heterodimerize. Strikingly, the structural interaction is tissue specific: nht did not interact with dTAF12 and dTAF4 did not interact with rye in a bacterial co-expression assay. We propose that the products of the five Drosophila genes encoding testis TAF homologs collaborate in an alternative TAF-containing protein complex to regulate a testis-specific gene expression program in primary spermatocytes required for terminal differentiation of male germ cells.

Prevalence, care, and outcomes for patients with diet-controlled diabetes in general practice: cross sectional survey.

BACKGROUND: Tight glycaemic control reduces microvascular complications in patients with type 1 and type 2 diabetes. We aimed to establish the proportion with type 2 diabetes treated through diet only and to determine levels of complications and quality of care received compared with patients on hypoglycaemic medication. METHODS: We undertook a cross-sectional study of 7870 patients with type 2 diabetes from a population of 253,618 patients from 42 general practices in the UK. Our primary outcome was process of care measures and diabetes-related complications. FINDINGS: 31.3% of all patients with type 2 diabetes are being managed with diet only (1% of the total population). More than four-fold variation between practices exists (range 15.6-73.2%). Patients treated with diet only are much less likely to have HbA1c (glycosylated haemoglobin) measurements, blood pressure, cholesterol, smoking, microalbuminuria testing, or screening for foot pulses recorded. 38.4% of patients with type 2 diabetes on medication have a HbA(1c) above 7.5% compared with 17.3% of those treated with diet only. Compared with those on medication, patients treated by diet only are more likely to have raised blood pressure and less likely to be on anti-hypertensive medication; they are 45% more likely to have raised cholesterol and less likely to be prescribed lipid-lowering medication. Although fewer of those treated by diet (68%) have diabetes-related complications compared with those on medication (80%), the rate is much higher than for the population without diabetes. INTERPRETATION: Diabetics treated by diet only have significant rates of complications and are less likely than those on medication to be adequately monitored. There is great scope for improved management within general practice.