Verocytotoxin Escherichia coli-Associated Haemolytic Uraemic Syndrome.

Aims To describe laboratory data on clinical human Verotoxigenic E. coli (VTEC) strains causing haemolytic uraemic syndrome (HUS) and to characterise the VTEC strains, thus contributing to risk mitigation to decrease HUS incidence in Ireland. Methods Laboratory characterisation was performed on isolates from 52 VTEC-associated HUS cases identified in the National clinical VTEC Reference Laboratory (NRL-VTEC) for the years 2012-2014. Data were analysed with respect to age, gender, serogroup and verotoxin type and subtype. Results 52/83 (62.6%) culture positive HUS cases were identified from laboratory data; 30 (57.7%) cases occurred in females. Seven HUS-associated serogroups and eleven patterns of verotoxin subtypes are described. Conclusion Ireland has the highest incidence of VTEC infection in Europe and a variety of VTEC serogroups causing clinical infection, suggesting any viable VTEC may potentially cause HUS. A broad diagnostic approach, to detect uncommon serotypes, should be considered when analysing clinical and food samples for VTEC.

Verotoxigenic Escherichia coli transmission in Ireland: a review of notified outbreaks, 2004-2012.

Verotoxigenic Escherichia coli (VTEC) are significant for their low infectious dose, their potential clinical severity and the frequency with which they generate outbreaks. To describe the relative importance of different outbreak transmission routes for VTEC infection in Ireland, we reviewed outbreak notification data for the period 2004-2012, describing the burden and characteristics of foodborne, waterborne, animal contact and person-to-person outbreaks. Outbreaks where person-to-person spread was reported as the sole transmission route accounted for more than half of all outbreaks and outbreaks cases, most notably in childcare facilities. The next most significant transmission route was waterborne spread from untreated or poorly treated private water supplies. The focus for reducing incidence of VTEC should be on reducing waterborne and person-to-person transmission, by publicizing Health Service Executive materials developed for consumers on private well management, and for childcare facility managers and public health professionals on prevention of person-to-person spread.

TPM3 deletions cause a hypercontractile congenital muscle stiffness phenotype.

OBJECTIVE: Mutations in TPM3, encoding Tpm3.12, cause a clinically and histopathologically diverse group of myopathies characterized by muscle weakness. We report two patients with novel de novo Tpm3.12 single glutamic acid deletions at positions ΔE218 and ΔE224, resulting in a significant hypercontractile phenotype with congenital muscle stiffness, rather than weakness, and respiratory failure in one patient. METHODS: The effect of the Tpm3.12 deletions on the contractile properties in dissected patient myofibers was measured. We used quantitative in vitro motility assay to measure Ca(2+) sensitivity of thin filaments reconstituted with recombinant Tpm3.12 ΔE218 and ΔE224. RESULTS: Contractility studies on permeabilized myofibers demonstrated reduced maximal active tension from both patients with increased Ca(2+) sensitivity and altered cross-bridge cycling kinetics in ΔE224 fibers. In vitro motility studies showed a two-fold increase in Ca(2+) sensitivity of the fraction of filaments motile and the filament sliding velocity concentrations for both mutations. INTERPRETATION: These data indicate that Tpm3.12 deletions ΔE218 and ΔE224 result in increased Ca(2+) sensitivity of the troponin-tropomyosin complex, resulting in abnormally active interaction of the actin and myosin complex. Both mutations are located in the charged motifs of the actin-binding residues of tropomyosin 3, thus disrupting the electrostatic interactions that facilitate accurate tropomyosin binding with actin necessary to prevent the on-state. The mutations destabilize the off-state and result in excessively sensitized excitation-contraction coupling of the contractile apparatus. This work expands the phenotypic spectrum of TPM3-related disease and provides insights into the pathophysiological mechanisms of the actin-tropomyosin complex.

Infant botulism due to C. butyricum type E toxin: a novel environmental association with pet terrapins.

We describe two cases of infant botulism due to Clostridium butyricum producing botulinum type E neurotoxin (BoNT/E) and a previously unreported environmental source. The infants presented at age 11 days with poor feeding and lethargy, hypotonia, dilated pupils and absent reflexes. Faecal samples were positive for C. butyricum BoNT/E. The infants recovered after treatment including botulism immune globulin intravenous (BIG-IV). C. butyricum BoNT/E was isolated from water from tanks housing pet 'yellow-bellied' terrapins (Trachemys scripta scripta): in case A the terrapins were in the infant's home; in case B a relative fed the terrapin prior to holding and feeding the infant when both visited another relative. C. butyricum isolates from the infants and the respective terrapin tank waters were indistinguishable by molecular typing. Review of a case of C. butyricum BoNT/E botulism in the UK found that there was a pet terrapin where the infant was living. It is concluded that the C. butyricum-producing BoNT type E in these cases of infant botulism most likely originated from pet terrapins. These findings reinforce public health advice that reptiles, including terrapins, are not suitable pets for children aged <5 years, and highlight the importance of hand washing after handling these pets.

Occurrence and characteristics of fastidious Campylobacteraceae species in porcine samples.

This study investigated the prevalence and characteristics of Campylobacteraceae including a range of fastidious species in porcine samples. Over a thirteen month period caecal contents (n=402) and pork carcass swabs (n=401) were collected from three pork abattoirs and pork products (n=399) were purchased at point of sale in the Republic of Ireland. Campylobacteraceae isolates were recovered by enrichment, membrane filtration and incubation in antibiotic free media under a modified atmosphere (3% O2, 5% H2, 10% CO2 and 82% N2). Campylobacteraceae isolates were identified as either genus Campylobacter or Arcobacter and then selected species were identified by Polymerase Chain Reaction (PCR). Campylobacteraceae were isolated from 103 (26%) caecal samples, 42 (10%) carcass swabs, and 59 (15%) pork products. Campylobacter coli was the most commonly isolated species found in (37%) all sample types but many fastidious species were also isolated including Campylobacter concisus (10%), Arcobacter butzleri (8%), Campylobacter helveticus (8%), Campylobacter mucosalis (6%), Arcobacter cryaerophilus (3%), Campylobacter fetus subsp. fetus (1%), Campylobacter jejuni subsp. jejuni (1%), Campylobacter lari (0.5%), Campylobacter curvus (0.5%) and Arcobacter skirrowii (0.5%). Among all isolates, 83% contained cadF and 98% flaA. In this study 35% of porcine C. coli were resistant to ciprofloxacin but none of the fastidious species demonstrated any resistance to this drug. The level of resistance to erythromycin was very high (up to 100%) in C. concisus and C. helveticus and this is a real concern as this is the current empiric drug of choice for treatment of severe gastroenteritic Campylobacter infections. The study shows that there is a much wider range of fastidious Campylobacteraceae present in porcine samples than previously assumed with C. concisus the second most common species isolated. The majority of fastidious Campylobacteraceae isolates obtained contained virulence genes and antibiotic resistance indicating potential public health significance.

Organically bound tritium (OBT) formation in rainbow trout (Oncorhynchus mykiss): HTO and OBT-spiked food exposure experiments.

In order to determine the rate of organically bound tritium (OBT) formation, rainbow trout (Oncorhynchus mykiss) were exposed to tritiated water (HTO) or OBT-spiked food. The HTO (in water) exposure study was conducted using a tritium activity concentration of approximately 7000 Bq/L and the OBT (in food) exposure study was conducted using a tritium activity concentration of approximately 30,000 Bq/L. Fish in both studies were expected to be exposed to similar tritium levels assuming 25% incorporation of the tritiated amino acids found in the food. Four different sampling campaigns of HTO exposure (Day 10, 30, 70, 140) and five different sampling campaigns of OBT-spiked food exposure (Day 9, 30, 70, 100, 140) were conducted to measure HTO and OBT activity concentrations in fish tissues. OBT depuration was also evaluated over a period of 30 days following the 140 d exposure studies. The results suggested that the OBT formation rate was slower when the fish were exposed to HTO compared to when the fish were ingesting OBT. In addition, the results indicated that OBT can bioaccumulate in fish tissues following OBT-spiked food exposure.

The effects of chronic tinnitus caused by acoustic trauma on social behaviour and anxiety in rats.

Tinnitus is associated with significant increases in anxiety disorders in humans, which is thought to affect social interaction; however, there has been only one previous study of the effects of tinnitus on social interaction in animals treated with salicylate and no previous study of the effects of tinnitus on anxiety in animals. In the present study, we used acoustic trauma to induce tinnitus in rats and investigated its effects on social interaction and anxiety in animals confirmed to have tinnitus. When social behaviours were grouped, we found that animals with tinnitus engaged in significantly more aggressive behaviours toward both tinnitus and sham control animals (P=0.03). When individual social behaviours were analysed without considering whether a tinnitus or sham animal was interacting with a member of its own treatment group, tinnitus animals were found to engage in significantly more anogenital investigation (P=0.01) and significantly less social grooming (P=0.003). When the data were analysed according to whether an animal was interacting with a member of its own group, tinnitus animals were found to bite sham animals significantly more than other tinnitus animals (P=0.005). Sham animals also bit tinnitus animals significantly more than other sham animals (P=0.02), as well as climbing away from them more (P=0.04), kicking (P=0.003), nudging them more (P=0.04), and sleeping with them more (P=0.02). By contrast, sham animals sniffed tinnitus animals significantly less than sham animals (P=0.05). There were no significant differences between the sham and tinnitus animals in performance in the elevated plus and elevated T maze tests of anxiety. However, tinnitus animals displayed a slight but significant increase in locomotor activity in the open field (P=0.04). These data suggest that tinnitus results in complex changes in social interaction in rats, which are not due simply to increases in anxiety.

Acoustic trauma that can cause tinnitus impairs impulsive control but not performance accuracy in the 5-choice serial reaction time task in rats.

Although tinnitus is an auditory disorder, it is often associated with attentional and emotional problems. Functional neuroimaging studies in humans have revealed that the hippocampus, amygdala and anterior cingulate, areas of the brain involved in emotion, attention and spatial processing, are also involved in auditory memory and tinnitus perception. However, few studies of tinnitus-evoked emotional and cognitive changes have been reported using animal models of tinnitus. In the present study, we investigated whether acoustic trauma that could cause tinnitus would affect attention and impulsivity in rats. Eight male Wistar rats were exposed to unilateral acoustic trauma (110 dB, 16 kHz for 1 h under anaesthesia) and eight rats underwent the same anaesthesia without acoustic trauma. Tinnitus was tested in noise-exposed rats using a frequency-specific shift in a discrimination function with a conditioned lick suppression paradigm. At 4 months after the noise exposure, the rats were tested in a 5-choice serial reaction time task. The behavioural procedure involved training the rats to discriminate a brief visual stimulus presented randomly in one of the five spatial locations and responding by poking its nose through the illuminated hole and collecting a food pellet from the magazine. While all of the animals performed equally well in making correct responses, the animals exposed to acoustic trauma made significantly more premature responses. The results suggest that rats exposed to acoustic trauma and some of which have chronic tinnitus are impaired in impulsive control, but not performance accuracy.

Comparison of disc diffusion and epsilometer (E-test) testing techniques to determine antimicrobial susceptibility of Campylobacter isolates of food and human clinical origin.

The antibiotic resistance profiles of 75 Campylobacter isolates of food and human clinical origin was determined by two agar diffusion susceptibility methods; disc diffusion and epsilometer-test (E-test). The most common therapeutic antimicrobials, erythromycin, ciprofloxacin and tetracycline were studied, along with chloramphenicol, ampicillin and naladixic acid. The resistance observed for each antimicrobial, as determined by both of methods, were statistically compared using Fisher two-tailed analysis. Of the six antimicrobials studied only two were shown to have statistically different patterns when resistance was compared by disc diffusion and E-test. The percentage of isolates resistant to clinically relevant antimicrobials using both techniques ranged from 6.6 to 21.3% for erythromycin, 25.3-26.6% for tetracycline and 33.3-36.0% for ciprofloxacin. The prevalence of multi-drug resistant (MDR) campylobacters (isolates resistant to 2 or more antimicrobials) for both disc diffusion and E-test was 44%. It can be concluded that, for four of the six antimicrobials assessed, antimicrobial resistance prevalences could be equally determined by either of the methods studied.

Molecular characterization of Irish E. coli O157:H7 isolates of human, bovine, ovine and porcine origin.

AIMS: To determine the degree of relatedness between isolates of Escherichia coli O157:H7 of human, bovine, ovine and porcine origin. METHODS AND RESULTS: Escherichia coli O157:H7 isolates were compared using (i) PFGE XbaI patterns, (ii) PCR profiles of virulence genes and (iii) the DNA sequences of genes reported to play a role in pathogenicity. The 77 E. coli O157:H7 isolates demonstrated 49 different PFGE patterns of which, eight were common to multiple isolates, and the remaining 41 were distinct. Isolates of different origin did not correlate, except for one cluster consisting of two human and two beef isolates. The majority of animal isolates had the same PCR profiles of virulence genes as those isolated from clinical patients. Single nucleotide polymorphisms (SNPs) were identified in the sequence of a 255-bp region of the vtx2 subunit A gene. CONCLUSIONS: Six SNPs were detected in the vtx2A gene, defining four different haplotypes. One nonsynonymous substitution encoded for an amino acid change from glutamic to aspartic acid. SIGNIFICANCE AND IMPACT OF THE STUDY: Results indicate that although E. coli O157:H7 isolates of differing origin were distinct by PFGE, the DNA sequences of the main virulence genes associated with human clinical illness were conserved.

Antibiotic resistance of retail food and human Campylobacter isolates on the island of Ireland from 2001-2002.

The antimicrobial resistance profiles of Campylobacter isolates recovered from a range of retail food samples (n=374) and humans (n=314) to eight antimicrobial compounds were investigated. High levels of resistance in food C. jejuni isolates were observed for ceftiofur (58%), ampicillin (25%) and nalidixic acid (17%) with lower levels observed for streptomycin (7.9%) and chloramphenicol (8.3%). A total of 80% of human C. jejuni isolates were resistant to ceftiofur, while 17% showed resistance to ampicillin and nalidixic acid, 8.6% to streptomycin and 4.1% to chloramphenicol. Resistance to clinically relevant antimicrobials such as erythromycin, ciprofloxacin and tetracycline was 6.7, 12, and 15% respectively for all food isolates and was similar to corresponding resistance prevalences observed for human isolates, where 6.4, 12 and 13% respectively were found to be resistant. Comparisons of C. jejuni isolates in each location showed a high degree of similarity although some regional variations did exist. Comparison of total C. jejuni and C. coli populations showed minor differences, with C. jejuni isolates more resistant to ampicillin and ceftiofur. Multidrug resistance patterns showed some profiles common to human and clinical isolates.

Haemolytic uraemic syndrome associated with interfamilial spread of E. coli O26:H11.

In September 2000, haemolytic uraemic syndrome (HUS) was diagnosed in a 10-month-old child with a prodromal history of vomiting and diarrhoea (non-bloody). Investigation revealed that a self-limiting gastrointestinal illness (mean duration 48 h) had occurred among immediate and extended family in the 2 weeks prior to the child's admission. The epidemiology of the illness suggested person-to-person spread. Five children (close family contacts) had E. coli O26 verocytotoxin (VT1 and VT2) isolated from stools. Stool culture and serology from the index case were negative for shiga toxin-producing E. coli (STEC) organisms. Control measures in accordance with the Public Health Laboratory Service (PHLS), verocytotoxogenic organisms (VTEC) guidelines were applied to prevent further spread among the extended family and contacts. Despite detailed food and environmental exposure histories, the source of the illness was not identified. This incident highlights the importance of investigation of cases of post-diarrhoeal HUS, for potential shiga toxin E. coli aetiology.

Occurrence of Campylobacter in retail foods in Ireland.

A surveillance study was carried out to determine the prevalence of Campylobacter in a range of retail foods purchased in three Irish cities over a 20-month period between March 2001 and October 2002. In total 2391 food samples were analysed during this period. Campylobacter was isolated from 444 raw chicken (49.9%), 33 turkey (37.5%) and 11 duck samples (45.8%). Lower isolation rates of 7/221 (3.2%), 10/197 (5.1%) and 31/262 (11.8%) were observed for raw beef, pork and lamb, respectively. One sample of pork paté from 120 samples analysed (0.8%) was Campylobacter-positive. A total of three shellfish samples (oysters) from 129 raw specimens examined (2.3%) were found to contain Campylobacter. Low prevalences of the organism (0.9%) were also isolated from fresh mushrooms. Of 62 raw bulk tank milk samples analysed, Campylobacter was recovered in a single sample (1.6%). Campylobacter was not detected in any of the comminuted pork puddings, prepared vegetables and salads, retail sandwiches or cheeses made from unpasteurised milk. In total, 543 Campylobacter were isolated from all of the food samples analysed, of which 453 (83.4%) were confirmed as Campylobacter jejuni and the remaining 90 (16.6%) as Campylobacter coli.

Nutrition in the community--an exploratory study of oral nutritional supplements in a health board area in Ireland.

BACKGROUND: A review of the General Medical (Payments) Scheme data in the Midland Health Board (MHB) Ireland identified a spend of just over euro 0.5 million on enteral nutritional supplements (oral and tube feeds) in an 11-month period in 1998 [General Medical Services (Payments) Board, 1998, MHB Clinical Nutritional Products: January-December 1998, Dublin]. In 2000, a figure of euro5 million was reported as the annual spend (oral and tube feeds) [General Medical Services (Payments) Board, 2000, MHB Clinical Nutritional Products: January-December 2000, Dublin]. Research has shown that a high proportion of Oral Nutritional Supplements (ONS) are inappropriately prescribed by primary care practitioners (Gall et al., 2001). The role of General Practitioners (GPs) and Public Health Nurses (PHNs) in prescribing ONS to patients aged 65 years and older was examined, as they are directly involved in the delivery of primary health care. AIM: (i) Assess current trends, decision-making processes and monitoring procedures in the use of ONS for older patients in the community. (ii) Identify whether nutritional assessments and appropriate nutritional criteria are standard practice in determining selection of ONS. METHODS: A study was conducted among 99 GPs and 120 PHNs in the MHB. All GPs were selected to participate and 50% (60) of PHNs were randomly selected. A telephone questionnaire was administered to each subject over a 2-week period. RESULTS: Both GPs (78%) and PHNs (47%) reported that their prescription of/recommendations for ONS had increased in the last 4 years. None conducted a full nutritional assessment, but 25% of PHNs used a Nutrition Screening Tool when trying to ascertain whether a patient requires an ONS. Only 19.6% of GPs and 6.8% of PHNs surveyed were aware of the calorie content of a standard 200 mL ONS (sip-feed). In addition, a very significant proportion of both GPs and PHNs do not appear to give appropriate dietary advice to patients who may be at risk of malnutrition. Only 55% of GPs stated that they would specifically review a patient's ONS prescription. All GPs said that they would not conduct a full nutritional assessment at the review appointment. CONCLUSION: The results of this study raise concerns as to the appropriateness of current ONS prescription and monitoring in the community. They also highlight the need for further intervention in the primary care setting in order to ensure that elderly malnourished patients are detected, treated and monitored in an appropriate and cost-effective manner.

A twelve-year review of central nervous system bacterial abscesses; presentation and aetiology.

OBJECTIVES: To review and document the changing patterns in diagnosis, causes and treatment of bacterial infections of the central nervous system (CNS) in a national neurosurgical unit only in patients from whom a specimen was obtained for culture. METHODS: The case notes, radiological results and laboratory records of all 163 patients in our institution who underwent a neurosurgical procedure between 1988 and 2000 for a CNS abscess in a national center were reviewed retrospectively. Those patients from whom there were no operative specimens (i.e. neurosurgical intervention was not performed) and who were treated empirically were excluded, as were patients with mycobacterial infection. RESULTS: The mean age of the 163 patients was 35.2 years. Headache, pyrexia and an altered mental state were the commonest presentations. The frontal lobe was the commonest anatomical site (62 patients, 38%) and the majority of abscesses occurred following community infections such as sinusitis and mastoiditis; no primary source could be identified in 32 (20%) patients. Bacteria were isolated from 73% of patients and polymicrobial infections occurred in 29 (17.7%) patients. Anaerobes accounted for only 13.6% of isolates and methicillin-resistant Staphylococcus aureus (MRSA) was isolated on five occasions, all in the last five years of this review. Sixteen (9.8%) patients died prior to discharge or transfer back to the original referring hospital and 18 (11%) patients developed epilepsy. CONCLUSION: There was a relatively high incidence of polymicrobial infection but the number of specimens with anaerobes was small, which may be because of the use of empiric metronidazole before surgical intervention. Most infections were community-acquired and responded well to a combination of surgical drainage and antibiotic therapy. The emergence of MRSA in this group of patients is, however, worrying.

Tube feeding controversial patients: what do dietitians think?

The use of tube feeding in some patients can be controversial, however, few studies have investigated dietitians' opinions on this subject. A cross-sectional survey of 345 members the Irish Nutrition and Dietetic Institute was conducted using a self-administered, anonymous, postal questionnaire. A 44% response rate was achieved. Mean number of years qualified was 9.3 (8.4). Eighty-one per cent of responders were involved in initiating tube feeding in stroke patients, and 8.5% in discontinuing tube feeding in a patient in a persistent vegetative state (PVS). Nine per cent felt that their input had no influence on the care plan of the patient with dementia and 67% felt that the information given to families (or other decision makers) concerning tube feeding was inadequate. The majority of respondents favoured tube feeding fictitious stroke and cancer patients, but less than half favoured tube feeding a fictitious patient in a PVS or a patient with dementia. When given similar scenarios involving themselves, fewer dietitians wanted to be tube fed.

Outbreak of Vero cytotoxin-producing Escherichia coli O157 in a child day care facility.

In December 1998, an outbreak of Vero cytotoxin-producing Escherichia coli (VTEC) O157 in a crèche affected ten out of 45 children and one out of five staff members. Eight cases were symptomatic and three were asymptomatic. There were two asymptomatic adult family contacts of child cases. All specimens were identified as VTEC O157:H7, phage type 32. None of the cases were seriously ill and none developed haemolytic uraemic syndrome (HUS). One child continued to excrete the organism for 14 weeks. The origin of the outbreak was not found but epidemiological investigation was suggestive of person-to-person spread. All children and staff were screened and excluded from the crèche until microbiological clearance was obtained. An inspection of the crèche revealed overcrowding and inadequacies in cleaning and in the food preparation facilities. These problems were remedied before children were re-admitted to the crèche. This outbreak demonstrates the ease with which VTEC O157 can be transmitted between small children. Two specific features of this outbreak were notable: (1) the mild self-limiting nature of the illness and (2) the prolonged shedding of the bacterium by one child.