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Context Intermittent rivers make up more than 50% of the world’s rivers. Refuge waterholes in these systems are critical for survival of fish populations. Aim This study investigated the effect that angling pressure imparts on native fish within refuge waterholes. Methods Golden perch (Macquaria ambigua) size structure was analysed in eight refuge waterholes across three catchments of the northern Murray–Darling Basin. Waterhole characteristics were recorded, with town proximity or remoteness being adopted as a surrogate variable for angling pressure. Key results Remoteness was found to be significantly correlated with median length, biomass, and proportion of legal-sized fish, but not with fish density. Sampling occurred after widespread connective flows, and the strength of remoteness as a predictor of median length suggests that redistribution of legal-sized golden perch on these flows was minimal. Legal-sized fish were poorly represented in the length–frequency distributions of near-town waterholes, and smaller size classes were well represented in most waterholes. Conclusion This study suggests that populations of golden perch in refuge waterholes near towns face significant angling pressure, with impacts on size structure at a local level. Implications These findings may have wider implications for the long-term resilience of the species.

To explore the experiences of adolescents with concussion and their parents who participated in a novel remotely delivered Tele-Active Rehabilitation (Tele-AR) intervention involving sub-symptom threshold exercise, education, and support. Specifically, we aimed to elicit perspectives regarding the remote delivery approach, valued aspects of the program, and perceived benefits of the intervention.This qualitative study took place within the context of a larger mixed-methods project exploring the feasibility of the Tele-AR intervention. A descriptive qualitative design was utilized. Semi-structured interviews were conducted with adolescents (n = 3; ages 14-17 years) with concussion and one of their parents (n = 3) within one week of completing the six-week Tele-AR intervention. Data were analyzed using thematic analysis.Four themes were identified that capture participant experiences in the intervention: (1) Enabling access to active rehabilitation; (2) Focusing on individual needs; (3) Learning to take responsibility for recovery; and (4) Convenience and comfort of engaging in rehabilitation from home.A small sample of 3 adolescents with concussion and their parents were satisfied with the Tele-AR intervention and appreciated the convenience and comfort of engaging in rehabilitation from home, which facilitated adolescents taking responsibility for their own recovery. Findings support continued study of Tele-AR, which may be an accessible intervention to facilitate recovery in adolescents with concussion.

This is the first report of betanodavirus infection in 2 species of finfish, Kuhlia rupestris (jungle perch) and Ambassis marianus (estuary perchlet). This report also describes isolation of betanodavirus from infected pond water using the SSN-1 cell line. Histopathology of K. rupestris larvae revealed vacuolation in the eye and brain, which was confirmed using betanodavirus-specific immunohistochemistry. The eye and brain from A. marianus and betanodavirus isolated from pond water were confirmed using real-time PCR and Sanger sequencing. High throughput sequencing was used to obtain betanodavirus sequences from paraffin blocks containing infected K. rupestris. The phylogenetic analysis of betanodavirus RNA1 and RNA2 sequences from all 3 sources were associated with the red-spotted grouper nervous necrosis virus (RGNNV) genotype. The RNA1 nucleotide sequence from jungle perch showed 100% identity with the betanodavirus water isolate and 99.37% identity with A. marianus. Furthermore, we have demonstrated the usefulness of combining recovery of viable virus from environmental samples through fish cell line culture with PCR testing as a means of validating the efficacy of chlorination to eradicate betanodavirus from the pond environment.

BACKGROUND Active rehabilitation involving subsymptom threshold exercise combined with education and support promotes recovery in youth with concussion but is typically delivered in person, which may limit accessibility for families because of a lack of services in their communities or logistical challenges to attending in-person sessions. OBJECTIVE This paper describes the evidence-based and theory-informed development of the Tele-Active Rehabilitation (Tele-AR) intervention for pediatric concussion, which was specifically designed for remote service delivery. METHODS The intervention was designed by clinician-researchers with experience in pediatric concussion rehabilitation following the Medical Research Council guidance for developing complex interventions. Development involved a critical review of the literature to identify existing evidence, the expansion of the theoretical basis for active rehabilitation, and the modeling of the intervention process and outcomes. RESULTS Tele-AR is a 6-week home exercise and education and support program facilitated through weekly videoconferencing appointments with a clinician. Exercise consists of low- to moderate-intensity subsymptom threshold aerobic activity and coordination drills that are individualized to participant needs and interests (prescribed for 3 days per week). Education includes the evidence-supported Concussion & You self-management program, which covers topics related to energy management, nutrition, hydration, sleep hygiene, and return to activity. Elements of self-determination theory are incorporated to support motivation and engagement. We present a logic model describing predicted intervention effects using a biopsychosocial conceptualization of outcomes after concussion. CONCLUSIONS The Tele-AR intervention may help to increase access to care that improves recovery and promotes a timely return to activity in youth with concussion. Future research is needed to evaluate the feasibility and efficacy of this approach. CLINICALTRIAL

Background Active rehabilitation involving subsymptom threshold exercise combined with education and support promotes recovery in youth with concussion but is typically delivered in person, which may limit accessibility for families because of a lack of services in their communities or logistical challenges to attending in-person sessions. Objective This paper describes the evidence-based and theory-informed development of the Tele-Active Rehabilitation (Tele-AR) intervention for pediatric concussion, which was specifically designed for remote service delivery. Methods The intervention was designed by clinician-researchers with experience in pediatric concussion rehabilitation following the Medical Research Council guidance for developing complex interventions. Development involved a critical review of the literature to identify existing evidence, the expansion of the theoretical basis for active rehabilitation, and the modeling of the intervention process and outcomes. Results Tele-AR is a 6-week home exercise and education and support program facilitated through weekly videoconferencing appointments with a clinician. Exercise consists of low- to moderate-intensity subsymptom threshold aerobic activity and coordination drills that are individualized to participant needs and interests (prescribed for 3 days per week). Education includes the evidence-supported Concussion & You self-management program, which covers topics related to energy management, nutrition, hydration, sleep hygiene, and return to activity. Elements of self-determination theory are incorporated to support motivation and engagement. We present a logic model describing predicted intervention effects using a biopsychosocial conceptualization of outcomes after concussion. Conclusions The Tele-AR intervention may help to increase access to care that improves recovery and promotes a timely return to activity in youth with concussion. Future research is needed to evaluate the feasibility and efficacy of this approach.

Degradation of instream habitats in the northern Murray–Darling Basin has occurred through numerous stressors, including siltation, clearing of bankside vegetation, intrusion of livestock and impacts of pest species. A better understanding of habitat preferences of native fish species could help guide future instream habitat restoration actions. The habitat choices of seven native fish species, juvenile Murray Cod (Maccullochella peelii), juvenile Golden Perch (Macquaria ambigua ambigua), juvenile Silver Perch (Bidyanus bidyanus), adult Murray–Darling Rainbowfish (Melanotaenia fluviatilis), adult Olive Perchlet (Ambassis agassizii), adult Un‐specked Hardyhead (Craterocephalus stercusmuscarum fulvus) and adult carp gudgeons (Hypseleotris spp.) were tested in preference troughs to help inform potential habitat restoration actions in the Condamine catchment. Each species was given a choice between pair combinations of open sandy habitat, submerged macrophytes, emergent plants and rocky rubble. Habitat preferences varied between species. Murray Cod, Golden Perch, carp gudgeons and Olive Perchlets preferred structure over open sandy habitat, whilst juvenile Silver Perch, Un‐specked Hardyhead and Murray–Darling Rainbowfish did not avoid open sandy habitats. Juvenile Murray Cod preferred rocky rubble habitat over all other habitat choices. Use of complex rock piles to provide nursery habitat for Murray Cod populations is a potential restoration option. Introduction of rock could also benefit Golden Perch and carp gudgeons. Use of emergent plants, submerged macrophytes and rocky rubble for habitat restoration all appear to have merit for one or more species of small‐bodied fishes or juvenile stages of larger sized fishes. Rocky rubble or floating attached macrophytes could be viable restoration options in areas too turbid to establish submerged macrophytes. These habitat interventions would complement existing actions such as re‐snagging and provision of fish passage to assist with sustainable management of native fish populations.

Because of concerns regarding the prevalence of Campylobacter in chicken meat, member countries in the European Union (EU) undertook a surveillance program to determine Campylobacter levels on chicken carcasses. A sample mass of 25 g principally composed of neck skin was used, although breast skin could also be used if there were insufficient neck skin to meet the required sample mass. The aim was to establish a baseline for Campylobacter contamination of carcasses against which future interventions could be assessed. However, in the United Kingdom researchers wanted to explore whether different ratios of neck to breast skin in samples could affect the results obtained. A comparison of the Campylobacter levels on neck and breast skin samples obtained from the same chilled chicken carcasses was undertaken at four chicken slaughterhouses. The neck skins were significantly more heavily contaminated (P < 0.05) with Campylobacter than was breast skin. Statistical analyses revealed no relationship that would allow a conversion between levels obtained on the two skin types. Ongoing surveillance of Campylobacter for 6 years was funded by United Kingdom poultry processors using samples consisting solely of neck skin, and the results of this surveillance (2011 to 2016) are reported here. Given the higher Campylobacter levels on a sample exclusively consisting of neck skin, this protocol could yield results with which the poultry industry would find it more difficult to achieve the contamination reduction target. The contamination reduction target for the United Kingdom (≤10% of chicken carcasses exceeding neck skin Campylobacter levels of 1,000 CFU/g) was not met by the end of 2015, the date stipulated by the United Kingdom government.

peer-reviewed Listeriosis is a foodborne disease, with a high mortality rate, that predominantly affects the elderly. Under European Union legislation, EC 2073/2005, food business operators are encouraged to undertake sampling to ensure that the food processing environment, and required to ensure that food products, are free of Listeria monocytogenes. To determine the prevalence of L. monocytogenes in smaller food processing facilities in Northern Ireland, 24 companies submitted six processing environment swabs and two food samples every two months for eighteen months (July 2015 to November 2016) for L. monocytogenes examination. The prevalence of L. monocytogenes was 4.6% in food samples, and 6.3% in processing environment swabs. Over the duration of the study, 96 isolates of L. monocytogenes were obtained, one from each positive sample, except for two meat samples that had >100 cfu/g, where two isolates were obtained from each sample. No seasonality in occurrence of L. monocytogenes was seen for food isolates but significantly higher numbers of positive processing environment swabs were found in the warmer months of May, July and September (p = .007). Pulsed Field Gel Electrophoresis (PFGE) analysis revealed the presence of 27 pulsotypes; 9 pulsotypes were shared between different facilities and 9 were persistent. Based on a Combase predictive growth model, 77.5% (n = 130) of the foods tested were predicted to support the growth of L. monocytogenes. All of the isolates carried the pathogenicity genes inlA and actA and 71.4% carried qacH, which confers resistance to quaternary ammonium compounds which are frequently used in sanitizers. Whole genome sequencing of the isolates allowed multi-locus sequence typing to be undertaken. The data indicated that the sequence types identified included those with disease-causing ability, highlighting the disease-causing potential of the isolates.

Aims. To identify production and processing practices that might reduce Campylobacter numbers contaminating chicken broiler carcasses.Methods and Results. Numbers of campylobacters were determined on carcass neck skins after processing or in broiler house litter samples. Supplementary information that described farm layouts, farming conditions for individual flocks, the slaughterhouse layouts and operating conditions inside plants was collected, matched with each Campylobacter test result. Statistical models predicting the numbers of campylobacters on neck skins and in litter were constructed. Carcass microbial contamination was more strongly influenced by on farm production practices compared with slaughterhouse activities. We observed correlations between the chilling, washing and de-feathering stages of processing and numbers of campylobacters on carcasses. There were factors on farm that also correlated with numbers of campylobacters in litter. These included bird gender, the exclusion of dogs from houses, beetle presence in the house litter and the materials used to construct the house frame.Conclusions. Changes to farming practices have greater potential for reducing chicken carcass microbial contamination compared with processing interventions.Significance and impact of the study. Routine commercial practices were identified that were correlated with lowered numbers of campylobacters. Consequently, these practices are likely to be both cost-effective and suitable for adoption into established farms and commercial processing.

Common carp (Cyprinus carpio) are one of the world's most destructive vertebrate pests. In Australia, they dominate many aquatic ecosystems causing a severe threat to aquatic plants, invertebrates, water quality, native fish and social amenity. The Australian Government is considering release of cyprinid herpesvirus-3 (CyHV-3) as a control measure and consequently a robust, continental-scale estimate of the carp population and biomass is essential to inform planning and risk management. Here, we pioneer a novel model-based approach to provide the first estimate of carp density (no/ha) and biomass density (kg/ha) at river reach/waterbody, basin and continental scales. We built a spatial layer of rivers and waterbodies, classified aquatic habitats and calculated the area of each throughout the range of carp in Australia. We then developed a database of fishery-independent electrofishing catch-per-unit-effort (CPUE) for habitat types, containing catch information for 574,145 carp caught at 4831 sites. Eastern Australia accounted for 96% of carp biomass and 92% of the total available wetted habitat area (16,686 km2) was occupied. To correct these data for variable detection efficiencies, we used existing electrofishing data and undertook additional field experiments to establish relationships between relative and absolute abundances. We then scaled-up site-based estimates to habitat types to generate continental estimates. The number of carp was estimated at 199.2 M (95%Crl: 106 M to 357.6 M) for an ‘average’ hydrological scenario and 357.5 M (95%Crl: 178.9 M to 685.1 M) for a ‘wet’ hydrological scenario. In eastern Australia, these numbers correspond with biomasses of 205,774 t (95%Crl: 117,532–356,482 t) (average scenario) and 368,357 t (95%Crl: 184,234–705,630 t) (wet scenario). At a continental scale the total biomass was estimated at 215,456 t for an ‘average’ hydrological scenario. Perennial lowland rivers had the highest CPUE and greatest biomass density (up to 826 kg/ha) and the modelled biomass exceeded a density-impact threshold of 80–100 kg/ha in 54% of wetlands and 97% of stream area in large lowland rivers. The continental-scale biomass estimates provide a baseline for focusing national conservation strategies to reduce carp populations below thresholds needed to restore aquatic ecosystems at a range of spatial scales.

Objective To characterize the absorption, distribution, metabolism, and excretion of naloxegol, a PEGylated derivative of the µ-opioid antagonist naloxone, in healthy male subjects. Materials and methods [14C]-Labeled naloxegol (27 mg, 3.43 MBq) was administered as an oral solution to 6 fasted subjects. Blood, fecal, and urine samples were collected predose and at various intervals postdose. Naloxegol and its metabolites were quantified or identified by liquid chromatography with radiometric or mass spectrometric detection. Pharmacokinetic parameters were calculated for each subject, and metabolite identification was performed by liquid chromatography with parallel radioactivity measurement and mass spectrometry. Results Naloxegol was rapidly absorbed, with a maximum plasma concentration (geometric mean) of 51 ng/mL reached before 2 hours after dosing. A second peak in the observed naloxegol and [14C] plasma concentration-time profiles was observed at ~3 hours and was likely due to enterohepatic recycling of parent naloxegol. Distribution to red blood cells was negligible. Metabolism of [14C]-naloxegol was rapid and extensive and occurred via demethylation and oxidation, dealkylation, and shortening of the polyethylene glycol chain. Mean cumulative recovery of radioactivity was 84.2% of the total dose, with ~68.9% recovered within 96 hours of dosing. Fecal excretion was the predominant route of elimination, with mean recoveries of total radioactivity in feces and urine of 67.7% and 16.0%, respectively. Unchanged naloxegol accounted for ~1/4 of the radioactivity recovered in feces. Conclusions Naloxegol was rapidly absorbed and cleared via metabolism, with predominantly fecal excretion of parent and metabolites.

This study devised a protocol for the manufacture of commercial quantities of chicken liver pâté that reliably destroyed campylobacters. A literature search identified 40 pâté manufacture recipes. Recipes stages with a potential to be antimicrobial were assembled to form a new protocol that included washing with organic acid, freeze-thaw and flambé in alcohol. Naturally-contaminated, high-risk livers were obtained from clearance flocks at slaughter and the effect of each stage of the protocol on Campylobacter populations was determined. Organic acid washing changed the color of the liver surfaces. However, there were no significant differences between liver surface color changes when a range of concentrations of lactic acid and ethanoic acid washes were compared by reflective spectrophotometry. A 5% (w/v) acid wash reduced numbers of indigenous campylobacters by around 1.5 log10 CFU/g for both acids. The use of a Bain Marie was found to more reproducibly apply heat compared with pan-frying. Antimicrobial recipe stages reduced the numbers of campylobacters, but not significantly if thermal processing was ineffective. Cooking to 63°C was confirmed to be a critical control point for campylobacters cooked in a Bain Marie. Organoleptic and sensory assessment of pâté determined an overall preference for pâté made from frozen livers.

Aims: To determine the fate of E. coli on vegetables that were processed through commercial wash treatments and stored under simulated retail conditions at 4oC or wholesale at fluctuating ambient temperatures (0-25oC, dependent on season). Methods and Results: Bovine slurry that was naturally contaminated with Escherichia coli O145 was applied without dilution or diluted 1/10 using borehole water to growing potatoes, leeks or carrots. Manure was applied one week prior to harvest to simulate a near-harvest contamination event by manure deposition or an application of contaminated water to simulate a flooding event or irrigation from a contaminated water source. At harvest, crops were contaminated at up to two log cfu/g. Washing transferred E. coli into the water of a flotation tank used for potato washing and did not completely remove all traces of contamination from the crop. Manure contaminated potatoes were observed to contain 0.72 cfu E. coli O145/g after processing and retail storage. Manure-contaminated leeks harboured 0.73 – 1.55 cfu E. coli O145/g after washing and storage. There was no cross contamination when leeks were spray-washed. Washing in an abrasive drum resulted in less than perfect decontamination for manure-contaminated carrots. There were five post-distribution isolations from carrots irrigated with contaminated water 24h prior to harvest. Conclusions: Standard commercial washing and distribution conditions may be insufficient to reliably control human pathogenic E. coli on fresh produce. Significance and Impact: Previous speculation that the cause of a UK foodborne disease outbreak was soil from imperfectly cleaned vegetables is plausible.

Many freshwater fishes are imperilled globally, and there is a need for easily accessible, contemporary ecological knowledge to guide management. This compendium contains knowledge collated from over 600 publications and 27 expert workshops to support the restoration of 9 priority native freshwater fish species, representative of the range of life-history strategies and values in south-eastern Australia’s Murray–Darling Basin. To help prioritise future research investment and restoration actions, ecological knowledge and threats were assessed for each species and life stage. There is considerable new knowledge (80% of publications used were from the past 20 years), but this varied among species and life stages, with most known about adults, then egg, juvenile and larval stages (in that order). The biggest knowledge gaps concerned early life stage requirements, survival, recruitment, growth rates, condition and movements. Key threats include reduced longitudinal and lateral connectivity, altered flows, loss of refugia, reductions in both flowing (lotic) and slackwater riverine habitats, degradation of wetland habitats, alien species interactions and loss of aquatic vegetation. Examples and case studies illustrating the application of this knowledge to underpin effective restoration management are provided. This extensive ecological evidence base for multiple species is presented in a tabular format to assist a range of readers.

This study was conducted to analyse the current practices used by the Scottish smoked salmon industry that will affect the likelihood of Listeria monocytogenes contamination in products. Sixteen visits to smoked salmon premises were conducted between June and November 2011, interviews were carried out based on a questionnaire. The results indicate that most processors carry out appropriate food safety practices, but some improvements are needed in order to minimize the risk of Listeria contamination. It was found that the larger processors achieved better temperature control than the smaller processors. Approximately half of the visited premises needed to improve their refrigerated storage. The risk of ceiling condensation dripping onto product was a common problem, but the smaller premises were the most affected. Small food business operators require additional information on how cleaning and sanitation throughout the process can reduce contamination of the final product. Furthermore, guidance describing the best way of determining shelf life was requested by small processors. Fifty six percent of the smoked salmon processors (mostly large and medium size) tested the product for L. monocytogenes and prevalence ranged widely (0–12%) between processors. Those processors having the highest Listeria prevalence were also those most concerned about what microbiological testing should be carried out and how to evaluate the quality of their products. Most processors rarely exceeded (i.e. once every several years) the statutory limit set by the European Union (>100 cfu/g or presence in 25 g). The small producers did not undertake product testing for Listeria because of high test costs and lack of technical expertise. Hence, it was concluded that sharing expertise between producers, especially to smaller processors would be beneficial in terms of consumer protection.

The aims of this study were (i) to determine the prevalence and numbers of campylobacters in 63 samples of raw livers purchased at retail across the UK and (ii) to investigate whether the freezing of chicken livers contaminated with Campylobacter was a reliable method for decontamination. Chicken livers naturally contaminated with campylobacters were subjected to freezing at -15 and -25°C for one day and 7 days. Numbers of campylobacters on the livers were determined immediately before and after a 24-h or 7-days freeze treatment and daily during 3 days post-thaw refrigerated storage. Freezing for 24 h at -25°C can reduce numbers of Campylobacter by up to 2 log10 CFU g(-1). Freezing the livers for 24 h at -25°C, thawing overnight in a fridge set to 4°C and refreezing for another 24 h at -25°C reduced the numbers of campylobacters by up to three logs. Reduction in the numbers of campylobacters was significantly greater following a second freeze treatment compared with a single freeze treatment.

As a consequence of recent outbreaks of food borne illness associated with consumption of fresh produce, there is increased scrutiny of the microbiological risks associated with the consumption of ready to eat produce within the UK retail sector This project was funded to summarise the UK industry approach to managing risk in fresh produce production and identify best practices and areas of risk. Copies of quality assurance scheme codes of practice for fresh produce growers were reviewed from a food safety viewpoint against the stipulations of Codex Alimentarius. In general, all of the guides require good practices conducive with the production of food that is free from human pathogens. A survey of growers was undertaken to assess compliance with these codes and to identify areas in the codes that growers found difficult to comply with or to implement. Growers find it difficult to risk assess water sources used for irrigation and water usage. Furthermore, the selection of bacterial indicators that were encountered was diverse and growers experienced difficulty in choosing appropriate indicators to test for, and in using the results of these tests as the basis of informed decisions. Large growers supplying the UK multiple retail chains are required by their customers to comply with one or more assurance code as a condition for the sale of their produce. Small growers that supply farmers’ markets and wholesalers are under little or no pressure from their customers to do the same. This disparity in supply chains suggest that moves to encourage wholesalers to comply with assurance schemes would help reduce the risks to UK consumers from the consumption of fresh produce.

The pharmacokinetics and biotransformation of midazolam were investigated following single oral doses of 0.1, 1 and 10 mg/kg to chimeric mice with humanised livers (PXB mice) and to severe combined immunodeficient (SCID) mice used as controls. Pharmacokinetic analysis, on whole blood, revealed rapid absorption of the administered midazolam with a higher C(max) in PXB compared to SCID. The exposure to 1'-hydroxymidazolam was approximately 14-fold greater than to midazolam in the SCID mice and close to equivalent in the PXB mice. The metabolism of midazolam in SCID mice was faster than in the PXB mice such that pharmacokinetic data for midazolam in SCID mice could not be generated from the lowest dose in these animals. Both oxidative and conjugative metabolic pathways were identified in the PXB mice. All the major circulating metabolites observed in humans; 1'-hydroxymidazolam, 4'-hydroxymidazolam, 1',4'-dihydroxymidazolam and 1'-hydroxymidazolam glucuronide, were detected in the blood of PXB mice. However, 4'-hydroxymidazolam and the 1'-hydroxymidazolam glucuronide were not detected in blood samples obtained from SCID mice. The midazolam metabolite profile in the PXB mouse was similar to that previously reported for human suggesting that the PXB mouse model can provide a model system for predicting circulating human metabolites.

Statutory microbiological test results were collected from British meat plants over a 4-year period from June 2002 to May 2006. A total of 49,074 carcass test results from 19,409 cattle, 14,706 sheep, and 14,959 pig swabs and 95,179 environmental test results from surface swabs were obtained. These test results were donated by 94 slaughterhouses, which process about two thirds of the British national annual throughput of cattle, sheep, and pig carcasses. The data were collectively analyzed to determine any historical trends for numbers of total aerobes and Enterobacteriaceae. Significant reductions were observed in the numbers of indicator organisms on carcasses for all three species between 2002 and 2006. Reductions were also observed for numbers of aerobes on environmental and food contact surfaces. There were seasonal differences in bacterial numbers isolated from carcasses. Cattle and sheep carcasses had significantly higher numbers of total aerobes and Enterobacteriaceae in late summer and early autumn, whereas numbers of total aerobes on pig carcasses were higher in winter. Bacterial numbers on environmental surfaces were not influenced by the month that the swab samples were collected. Possible reasons for the observed reductions in bacterial numbers on carcasses and surfaces and the implications for carcass testing for process control purposes are discussed.

A survey of a large number of UK abattoirs was conducted via a questionnaire designed to obtain information on (i) throughput and species slaughtered; (ii) construction materials used; (iii) use and type of bedding and (iv) details of cleaning/sanitation regimes. A representative group of abattoirs were selected on the basis of the responses to the questionnaire, and the lairage at these plants investigated through enumeration of Escherichia coli remaining after routine cleansing operations. The aim of this study was to enable identification of “common lairage practices” and to assess the general status of the lairage hygiene and effectiveness of lairage cleaning in commercial UK abattoirs. The study shows that microbial contamination often remains in UK lairage holding pens after routine cleaning operations. It would appear that there are significant differences in the effectiveness of lairage cleaning programmes at commercial abattoirs, and that the stun-box-roll-out areas are often cleaned to a better standard than the holding areas. As a result of persistence of microbial contamination in the lairage, there is a possible risk of foodborne pathogens persisting in the environment and potentially contaminating animals and carcasses processed on subsequent days.

Desmethyl-gefitinib is a major metabolite of gefitinib observed in human plasma at concentrations similar to those of gefitinib. The epidermal growth factor receptor (EGFR)-related inhibitory effects of gefitinib and desmethyl-gefitinib have been compared both in vitro, using enzyme kinase assays and tumour cell growth inhibition, and in vivo by assessment of tumour xenografts growth inhibition in the mouse. Both gefitinib (IC(50) = 0.022 microM) and its desmethyl metabolite (0.036 microM) inhibited subcellular EGFR tyrosine kinase activity with a similar potency and selectivity. However, desmethyl-gefitinib (IC(50) = 0.76 microM) was 15 times less active than gefitinib (0.049 microM) against EGF-stimulated KB cell growth in a whole cell assay. Following a preliminary pharmacokinetic study to compare apparent oral bioavailability, gefitinib (75 mg kg(-1)) and desmethyl-gefitinib (150 mg kg(-1)) were administered orally for 15 days to female nude mice bearing LoVo tumour xenografts. Tumour concentrations of gefitinib (AUC = 300 microg h g(-1)) were much higher than those of desmethyl-gefitinib (44.3 microg h g(-1)), although plasma concentrations of gefitinib (48.4 microg h ml(-1)) and desmethyl-gefitinib (39.0 microg h ml(-1)) were quite similar at these dose levels. Gefitinib produced significant tumour growth inhibition throughout the course of the study ultimately resulting in a 50% decrease (compared with controls) by day 15. In contrast, although present at comparable plasma levels, desmethyl-gefitinib had little effect on tumour growth and is, therefore, considered unlikely to contribute significantly to the therapeutic activity of gefitinib in the clinical situation.

The in vitro metabolism of [(14)C]-gefitinib (1-3 microM) was investigated using human liver microsomes and a range of expressed human cytochrome P450 enzymes, with particular focus on the formation of O-desmethyl-gefitinib (M523595), the major metabolite observed in human plasma. High-performance liquid chromatography with ultraviolet light, radiochemical and mass spectral analysis, together with the availability of authentic standards, enabled quantification and structural identification of metabolites. On incubation with pooled human liver microsomes, [(14)C]-gefitinib underwent rapid and extensive metabolism to a number of metabolites, although M523595 was only a minor microsomal product. Formation of most metabolites was markedly decreased by ketoconazole, but M523595 production was inhibited only by quinidine. Gefitinib was metabolized extensively by expressed CYP3A4, producing a similar range of metabolites to liver microsomes, but M523595 was not formed. CYP1A2, 2C9 and 2C19 produced no measurable metabolism of gefitinib, while CYP3A5 produced a range of metabolites similar to CYP3A4, but to a much lower degree. In contrast, CYP2D6 catalysed rapid and extensive metabolism of gefitinib to M523595. While formation of M523595 was CYP2D6 mediated, the overall metabolism of gefitinib was dependent primarily on CYP3A4, and this was not obviously diminished in liver microsomes from CYP2D6 poor metabolizers.

Objective The study examined whether cognitive functioning, history of concussion (HOC), and sex predicted risk of sport concussion. Design Retrospective study design. Predictive models were used to determine the predictive ability of each variable. Setting Canadian University. Participants 708 data observations from 701 varsity athletes (41.2% female), representing 14 sports. Assessment of risk factors Two measures of cognitive functioning (mean reaction time and throughput [speed and accuracy]) were assessed using the Automated Neuropsychological Assessment Metrics testing battery. Categorical and continuous measures of HOC, and sex, were examined. Outcome measures Occurrence of concussion after baseline testing. Main results HOC was a significant predictor for both sexes. For every previous concussion, the odds of sustaining another concussion increased by 1.5 (95% Confidence Interval [CI]: 1.1, 2.1 [females]; 1.2, 1.9 [males]). Females with a HOC had twice the odds of sustaining another concussion than those without a HOC (CI: 1.1, 4.0). For males, the odds were three times (CI: 1.7, 5.6). Cognitive functioning and sex were not meaningful predictors. Conclusions This study supports previous findings that HOC is a risk factor and suggests that pre-injury cognitive functioning is not a risk factor for sport concussion. Assessing HOC in all athletes prior to their competitive season provides information regarding their risk of future sport concussion. Thus, it is important for clinicians to record HOC, and to encourage athletes to report concussions to ensure an accurate HOC record. Pre-injury cognitive screening of athletes is not recommended for assessing risk of future concussion. Competing interests Sandhya Mylabathula, Lynda Mainwaring, and Michael Hutchison None. Doug Richards is the Medical Director at the MacIntosh Sports Medicine Clinic at the University of Toronto Paul Comper is a clinical neuropsychologist consultant with the NHLPA and a member of the Concussion Working Group, but neither were related to the current research

1. Paracetamol overdose remains the leading cause of acute liver failure in humans. This study was undertaken in cynomolgus monkeys to study the pharmacokinetics, metabolism and the potential for hepatotoxic insult from paracetamol administration as a possible model for human toxicity. 2. No adverse effects were observed for doses of up to 900 mg/kg/d for 14 d. Only minor sporadic increases in alanine aminotransferase, aspartate aminotransferase and glutamate dehydrogenase in a number of animals were observed, with no clear dose response. 3. Toxicokinetic analysis showed good plasma exposure, albeit with less than proportional rises in Cmax and AUC, with increasing dose. The Cmax values in monkey were up to 3.5 times those associated with human liver toxicity and the AUC approx. 1000 times those associated with liver enzyme changes in 31-44% of human subjects. 4. Metabolite profiling of urine by (1)H NMR spectroscopy revealed paracetamol and its glucuronide and sulphate metabolites. Glutathione-derived metabolites, e.g. the cysteinyl conjugate, were only present in very low concentrations whilst the mercapturate was not detected. 5. These in vivo observations demonstrated that the cynomolgus monkey is remarkably resistant to paracetamol-induced toxicity and a poor model for investigating paracetamol-related hepatotoxicity in humans.

The hepatic cytochrome P450 reductase null (HRN) mouse, which has no functional hepatic Cyp P450s, may represent a useful model for examining extra-hepatic P450-related oxidative metabolism. Here the pharmacokinetics and metabolic fate of midazolam, a drug known to undergo such extra-hepatic metabolism, have been investigated in the HRN mouse and compared with a phenotypically normal strain (C57BL/6J). In addition, the effects of co-administration of the pan-P450 inhibitor 1′-aminobenzotriazole (ABT) on the metabolic profile have been compared in both strains. Significant pharmacokinetic differences for midazolam were observed between the two strains of mice with the HRN mice showing lower circulating concentrations of 1′-hydroxymidazolam but higher concentrations of 1′-hydroxymidazolam- O -glucuronide. A significant increase in midazolam exposure was seen upon ABT exposure for both strains of mice, but no differences in the area under the concentration time curves (AUC) for the monitored metabolites were observed. Although oxidative metabolism of midazolam was not abolished, significant decreases in 1′-hydroxymidazolam formation ratios were observed for both strains of mice exposed to ABT. Metabolite profiling of blood and bile showed a number of qualitative and quantitative differences between HRN and normal mice. These differences in midazolam metabolism between the two strains of mice clearly demonstrate the role that liver P450 enzymes play in the murine metabolism of midazolam. The fate of the compound in the HRN mice shows the importance of extrahepatic metabolism and also showed that these mice appear to be more capable of forming circulating phase II glucuronides than the normal strain.

Drug-drug interactions (DDIs) may cause serious drug toxicity and delay development of candidate drugs. Screening using human liver microsomes and hepatocytes can help predict DDIs but do not always provide the degree of certainty required for confident progression of a candidate drug. Thus a suitable in vivo test system could be of great value. Here a Cyp2c knockout (KO) mouse was investigated for studying DDIs using midazolam (MDZ) a standard human CYP3A4 substrate and troleandomycin (TAO) a potent human CYP3A4 inhibitor. Pharmacokinetics (PK) and biotransformation of MDZ were investigated following dosing to Cyp2c KO and wild type mice before and after TAO treatment. The noteworthy differences in the metabolism of MDZ in Cyp2c KO compared to wild type mice confirms the important role that Cyp2c enzymes play in the murine metabolism of MDZ in vivo. The impact of Cyp3a inhibition produced a further increase in circulating MDZ concentrations in all individuals from both strains of mice though the impact of the elimination of the Cyp2c pathway in the KO mice on the AUC was less than perhaps expected. We have shown that TAO produces an increase in the MDZ concentration and a reduction in the 1'hydroxymidazolam/midazolam formation ratio but the expected difference in the magnitude of this effect between the wild type and the Cyp2c KO mice was not seen. The magnitude of the TAO effect was also smaller than is reported in humans. Hence further work is required before this animal model could be used to predict clinical interactions.

There are a number of different approaches to the assessment of dose proportionality, but most of these are directed toward hypothesis testing. The analysis of dose proportionality studies, however, requires estimation rather than significance testing in order that the pharmacokinetic and clinical significance of any nonproportionality can be assessed.The methods of statistical analysis of these studies have been reviewed. An empirical model relating the log of the pharmacokinetic parameter linearly to the log of the dose (the “power model”) provides a readily interpreted measure of the degree of nonproportionality. The potential utility of this model has been demonstrated using a number of observed dataseis from different drugs. If there is doubt over the goodness of fit of the power model then analysis of variance (after log transformation), together with pairwise comparisons between doses, provides a suitable alternative approach.

Background Vandetanib is an orally available inhibitor of vascular endothelial growth factor receptor 2 and epidermal growth factor receptor and is rearranged during transfection tyrosine kinase activity. Development has included studies in non–small cell lung cancer and other tumor types. Accurate elimination kinetics were not determined in patient studies, and so the current human volunteer studies were performed to derive detailed kinetic data. Objective The aim of this study was to investigate pharmacokinetics, metabolism, excretion, and elimination kinetics after single oral doses of vandetanib in healthy subjects. Methods Three studies were conducted. In Study A (n = 23), cohorts of 8 subjects were randomized to receive double-blind, ascending doses of vandetanib (300–1200 mg) or placebo (6:2). Study B had a crossover design; subjects (n = 16) received vandetanib 300 mg under fed and fasted conditions. In Study C, subjects (n = 4) received [ 14 C] vandetanib 800 mg. Blood samples were collected for pharmacokinetic analysis for up to 28 days after the dose (Studies A and B) and 42 days after the dose (Study C). Plasma (all studies) and urine (Study A only) samples were collected for determination of vandetanib concentrations. In Study C radioactivity was measured in plasma, blood, urine, and feces, and metabolites were identified chromatographically. Tolerability was evaluated by recording of adverse events, clinical chemistry, hematology and urinalysis parameters, vital signs, and ECGs (all studies). Results Study A: mean (SD) age 34.4 (6.9) years; 23/23 male; mean (SD; range) weight 80.6 (8.1; 62−97) kg. Study B: mean (SD) age 35.3 (8.4) years; 15/16 male; mean (SD; range) weight 80.7 (11.2; 57−100) kg. Study C: mean (SD) age 60.3 (7.4) years; 4/4 male; mean (SD; range) weight 78.0 (7.7l; 72−87) kg. Pharmacokinetic parameters were consistent across all studies (Studies A and C, vandetanib 800 mg: geometric mean CL/F, 13.1–13.3 L/h; geometric mean apparent volume of distribution at steady state [V SS /F], 3592–4103 L; mean t ½ , 215.8–246.6 hours). Vandetanib was absorbed and eliminated slowly after single oral doses. AUC 0–∞ and C max were not significantly affected by ingestion of food. Median (range) T max was 8 (3–18) hours after food and 6 (5–18) hours after fasting. In plasma, concentrations of total radioactivity were higher than vandetanib concentrations at all time points, indicating the presence of circulating metabolites. Unchanged vandetanib and 2 anticipated metabolites (N-desmethylvandetanib and vandetanib N-oxide) were detected in plasma, urine, and feces. A further trace minor metabolite (glucuronide conjugate) was found in urine and feces. Approximately two thirds of the dose was recovered in feces (44%) and urine (25%) over 21 days, underlining the importance of both routes of elimination. Adverse events were reported by all subjects in Study A apart from 2 at a vandetanib dose of 300 mg; 12/15 (80%) and 14/16 (88%) subjects who took vandetanib under fed and fasted conditions, respectively, in Study B; and 2/4 (50%) subjects in Study C. No serious adverse events were reported. Increasing doses of vandetanib, in Study A, were associated with variable increases in systolic and diastolic blood pressures and variable increases from baseline in QTc interval. Hematuria was reported by 3 subjects (vandetanib 300 mg) in Study A. Small but consistent increases from baseline in serum creatinine were noted in subjects who received vandetanib in these studies. No other clinically important changes were observed in clinical chemistry, hematology and urinalysis parameters, vital signs, and ECGs in any of the studies. Conclusions The pharmacokinetics of vandetanib after single oral doses to healthy subjects were defined and the metabolic pathway was proposed. Vandetanib was absorbed and eliminated slowly with a t ½ of ∼10 days after single oral doses. The extent of absorption was not significantly affected by the presence of food. Approximately two thirds of the dose was recovered in feces (44%) and urine (25%) over 21 days. Unchanged vandetanib and N-desmethyl and N-oxide metabolites were detected in plasma, urine, and feces. Vandetanib appeared to be was well tolerated in the populations studied.

In recent years China has faced an increasing trilemma¡Xhow to pursue an independent domestic monetary policy and limit exchange rate flexibility, while at the same time facing large and growing international capital flows. This paper analyzes the impact of the trilemma on China's monetary policy as the country liberalizes its goods and financial markets and integrates with the world economy. It shows how China has sought to insulate its reserve money from the effects of balance of payments inflows by sterilizing through the issuance of central bank liabilities. However, we report empirical results indicating that sterilization dropped precipitously in 2006 in the face of the ongoing massive buildup of international reserves, leading to a surge in reserve money growth. We estimate a vector error correction model linking the surge in China's reserve money to broad money, real GDP, and the price level. We use this model to explore the inflationary implications of different policy scenarios. Under a scenario of continued rapid reserve money growth (consistent with limited sterilization of foreign exchange reserve accumulation) and strong economic growth, the model predicts a rapid increase in inflation. A model simulation using an extension of the framework that incorporates recent increases in bank reserve requirements also implies a rapid rise in inflation. By contrast, model simulations incorporating a sharp slowdown in economic growth lead to less inflation pressure even with a substantial buildup in international reserves.

The in vitro metabolism of gefitinib was investigated by incubating [14C]-gefitinib, as well as M537194, M387783 and M523595 (the main metabolites of gefitinib observed in man), at a concentration of 100 microM with human liver microsomes (4 mg ml(-1)) for 120 min. These relatively high substrate and microsomal protein concentrations were used in an effort to generate sufficient quantities of metabolites for identification. HPLC with ultraviolet light, radiochemical and mass spectral analysis, together with the availability of authentic standards, enabled quantification and structural identification of a large number of metabolites. Although 16 metabolites were identified, metabolism was restricted to three regions of the molecule. The major pathway involved morpholine ring-opening and step-wise removal of the morpholine ring and propoxy side chain. O-demethylation of the quinazoline methoxy group was a quantitatively less important pathway, in contrast to the clinical situation, where O-desmethyl gefitinib (M523595) is the predominant plasma metabolite. The third metabolic route, oxidative defluorination, was only a minor route of metabolism. Some metabolites were formed by a combination of these processes, but no metabolism was observed in other parts of the molecule. Incubation of gefitinib produced ten identified metabolites, but the use of the three main in vivo metabolites as additional substrates enabled a more comprehensive metabolic pathway to be constructed and this has been valuable in supporting the more limited data available from the human in vivo study.

The pharmacokinetics of gefitinib and its metabolites in rat and dog were investigated in preclinical studies conducted to support the safety evaluation and clinical development of gefitinib, the first EGFR tyrosine kinase inhibitor approved for the treatment of non-small-cell lung cancer. Following intravenous dosing (5 mg kg(-1), gefitinib plasma half-life was 3-6h in rats and dogs, although studies using a more sensitive HPLC-MS assay produced longer estimates of half-life (7-14h). In these studies, plasma clearance was high (male rat: 25 ml min(-1) kg(-1); female rat: 16 ml min(-1) kg(-1); male dog: 16 ml min(-1) kg(-1)), as was the volume of distribution (8.0-10.41 kg(-1) in rat; 6.31 kg(-1) in dog), and exposure in female rats was double that in males. Following administration of [14C]-gefitinib, concentrations of radioactivity in plasma exceeded gefitinib throughout the profile, indicating the presence of circulating metabolites in both rat and dog. An HPLC-MS assay was developed to measure concentrations of gefitinib and five potential metabolites in plasma. All five metabolites were detected in the rat, but at levels much lower than gefitinib. In the dog, exposure to gefitinib and M523595 was similar, with much lower concentrations of M537194 and only trace levels of the other metabolites. This profile of metabolites is similar to that observed in man.

Following oral administration of [14C]-gefitinib to albino and pigmented rats, radioactivity was widely and rapidly distributed, with the highest levels being found in liver, kidney, lung and gastrointestinal tract, but with only low levels penetrating the brain. Levels of radioactivity persisted in melanin-containing tissues (pigmented eye and skin). Binding to plasma proteins was high (86-94%) across the range of species examined and was 91% in human plasma. Substantial binding occurred to both human serum albumin and alpha-1 acid glycoprotein. Following oral and intravenous administration of [14C]-gefitinib, excretion of radioactivity by rat, dog and human occurred predominantly via the bile into faeces, with < 7% of the dose being eliminated in urine. In all three species, gefitinib was cleared primarily by metabolism. In rat, morpholine ring oxidation was the major route of metabolism, leading to the formation of M537194 and M608236 as the main biliary metabolites. Morpholine ring oxidation, together with production of M523595 by O-demethylation of the quinazoline moiety, were the predominant pathways in dog, with oxidative defluorination also occurring to a lesser degree. Pathways in healthy human volunteers were similar to dog, with O-demethylation and morpholine ring oxidation representing the major routes of metabolism.

To assess the impact of intraaortic balloon counterpulsation on coronary, renal and aortic blood flow in an animal heart failure model.Heart failure exacerbations are still often treated with inotropic medications despite a lack of evidence demonstrating any benefit with these drugs. Intraaortic balloon counterpulsation may be considered in certain cases a bridge to recovery.Four juvenile pigs underwent pacemaker implantation to induce a rapid-pacing mediated dilated cardiomyopathy. After approximately 4 weeks of rapid pacing, the mean ejection fraction was reduced to 28.8+/-9.5% with a mean systolic blood pressure of 64/44 mmHg. The pigs then underwent surgical placement of flow probes around the circumflex coronary artery, renal artery and infrarenal aorta. A Millar catheter was used to calculate Dp/Dt and a Swan-Ganz to calculate cardiac output. Data were recorded at baseline and after 10 minutes of balloon pumping. The pigs were euthanized post-procedure.Coronary blood flow was increased 9.7% by balloon counterpulsation from 38.3 +/- 12.0 to 42.0+/-11.4 ml/s (p=NS). Renal blood flow was reduced 11.9% by counterpulsation from 130.0+/-88.6 ml/s to 114.5+/-76.6 ml/s (p=NS). Infrarenal aortic blood flow was not changed (mean of 900 ml/s with and without counterpulsation); blood pressure, cardiac output and Dp/Dt were not changed after 10 minutes of pumping. There was little impact observed by changing the position of the balloon closer to or farther away from the apex of the aortic arch.Intraaortic balloon counterpulsation did not significantly improve hemodynamics in the pig heart failure model. This may be attributed to the high compliance of the juvenile pig's aorta, thus attenuating the pressure wave generated by counterpulsation. A larger volume balloon would merit investigation for this application.

From immunohistochemical and ligand-binding studies, it is known that the epidermal growth factor receptor (EGFR), a member of the erbB family of receptors, is expressed in tissues of epithelial, mesenchymal and neuronal origin and plays a major role in normal cellular processes such as proliferation, differentiation and development. EGFR is highly expressed in a number of solid tumours and its expression correlates with tumour progression, resistance to chemotherapy and a poor prognosis; it is consequently an attractive target for the rational design of novel anticancer agents. Knowledge of the role of EGFR in normal tissues will help the understanding of the adverse events associated with such agents. Studies in knockout mice and preclinical toxicology studies have shown that the major effects of inhibiting the EGFR are skin and gastrointestinal toxicities. Clinical studies with inhibitors of EGFR, such as gefitinib, cetuximab and erlotinib, have shown a favourable adverse-event profile, primarily consisting of skin and gastrointestinal toxicities, as predicted from the mechanism-based effects observed in preclinical studies.