Measuring antimicrobial use on dairy farms: A method comparison cohort study.

Antimicrobial use on UK dairy farms is measured for surveillance purposes, with veterinary sales data as a proxy for use. Two other methods of recording use have been used commonly on-farm: medicine waste bins and farm medicine records. However, none of these methods has been validated to measure antimicrobial use. The objective of this research was to assess agreement between the 3 most common methods for measuring on-farm antimicrobial use with a predetermined reference method on UK dairy farms. Antimicrobial use was measured prospectively on 26 UK dairy farms using medicine waste bins into which participants placed all discarded medicine packaging for a 12-mo period. At the end of 12 mo, farm medicine records and veterinary sales data were obtained retrospectively for participating farms. The reference method used was based on pre- and poststudy inventories combined with veterinary sales data. We investigated the systematic difference between the mean on-farm antimicrobial use measured by each of the 3 methods and a reference method, using one-way repeated-measures ANOVA models. Reliability and clinical relevance of the agreement between each pair of methods was quantified using the concordance correlation coefficient (CCC) and the Bland-Altman method, respectively. When compared with the reference method, veterinary sales data had excellent reliability for injectable antimicrobials and intramammary antimicrobials [95% confidence interval (CI) of CCC > 0.90] and moderate to excellent reliability for other antimicrobials (95% CI of CCC: 0.68-0.97). Medicine waste bins had good to excellent reliability for injectable (95% CI of CCC: 0.84-0.99), and intramammary products (95% CI of CCC: 0.78-0.94) and no agreement for other forms of antimicrobial. Farm medicine records did not agree for any form of antimicrobial when compared with the reference method. The use of veterinary sales data as a proxy for on-farm antimicrobial use in the UK represented excellent statistical reliability and offered clinically good agreement with the reference method when used to measure injectable antimicrobials. This study applies to the UK context and included a relatively small number of farms. However, these results have research and policy implications, both nationally and internationally, and are essential in accurately quantifying agricultural antimicrobial use to inform both animal and human health.

Gut DYSBIOSIS and altered barrier function precedes the appearance of metabolic syndrome in a rat model of nutrient-induced catch-up growth.

Nutritional restriction early in life followed by catch-up growth has been associated with increased risk of metabolic syndrome in adulthood. To elucidate whether altered gut colonization underlies the mechanisms responsible of this predisposition gut microbiome was studied before or afterwards catch-up growth. Offspring of dams fed ad libitum (C) or undernourished during pregnancy and suckling (U), were weaned onto high-fat diet (HFD) for 22 weeks (CHF and UHF, respectively) or continued on their diet. HF-feeding induced glucose intolerance (P<.05), insulin resistance (P<.001), and white adipose tissue inflammation (P<.001) in UHF rats compared to CHF. Analyses of gut microbial composition before catch-up growth revealed reduced F/B ratio and significant expansion of the mucolytic genera Akkermansia (P<.05) and Desulfovibrio (P<.05) in U pups. Although relative abundance of Akkermansia remained elevated to adulthood in U rats, HFD normalized its levels to C and CHF. Food-restriction increased intestinal permeability causing disorganization on the tight-junction proteins of colonic epithelium, Zonula Occludens-1 (ZO-1) and occludin, and reducing the mucus thickness layer in U adult rats. The levels of ZO-1 and occludin were not recovered in U rats after HF-feeding. This event was correlated with increased circulating levels of bacterial lipopolysaccharides in both U and UHF adult rats. Even more, serum lipopolysaccharides were already elevated in U rats compared to C group (P<.001) at weaning. Thus, gut dysbiosis and chronic endotoxemia observed in U rats, even before catch-up growth, might anticipate a pro-inflammatory milieu promoting metabolic diseases when fed hyperlipidic diets.

New approaches to pharmacosurveillance for monitoring prescription frequency, diversity, and co-prescription in a large sentinel network of companion animal veterinary practices in the United Kingdom, 2014-2016.

Pharmaceutical agents (PAs) are commonly prescribed in companion animal practice in the United Kingdom. However, little is known about PA prescription on a population-level, particularly with respect to PAs authorised for human use alone prescribed via the veterinary cascade; this raises important questions regarding the efficacy and safety of PAs prescribed to companion animals. This study explored new approaches for describing PA prescription, diversity and co-prescription in dogs, cats and rabbits utilising electronic health records (EHRs) from a sentinel network of 457 companion animal-treating veterinary sites throughout the UK over a 2-year period (2014-2016). A novel text mining-based identification and classification methodology was utilised to semi-automatically map practitioner-defined product descriptions recorded in 918,333 EHRs from 413,870 dogs encompassing 1,242,270 prescriptions; 352,730 EHRs from 200,541 cats encompassing 491,554 prescriptions, and 22,526 EHRS from 13,398 rabbits encompassing 18,490 prescriptions respectively. PA prescription as a percentage of booked consultations was 65.4% (95% confidence interval, CI, 64.6-66.3) in dogs; in cats it was 69.1% (95% CI, 67.9-70.2) and in rabbits, 56.3% (95% CI, 54.7-57.8). Vaccines were the most commonly prescribed PAs in all three species, with antibiotics, antimycotics, and parasiticides also commonly prescribed. PA prescription utilising products authorised for human use only (hence, 'human-authorised') comprised 5.1% (95% CI, 4.7-5.5) of total canine prescription events; in cats it was 2.8% (95% CI, 2.6-3.0), and in rabbits, 7.8% (95% CI, 6.5-9.0). The most commonly prescribed human-authorised PA in dogs was metronidazole (antibiotic); in cats and rabbits it was ranitidine (H2 histamine receptor antagonist). Using a new approach utilising the Simpson's Diversity Index (an ecological measure of relative animal, plant etc. species abundance), we identified differences in prescription based on presenting complaint and species, with rabbits generally exposed to a less diverse range of PAs than dogs or cats, potentially reflecting the paucity of authorised PAs for use in rabbits. Finally, through a novel application of network analysis, we demonstrated the existence of three major co-prescription groups (preventive health; treatment of disease, and euthanasia); a trend commonly observed in practice. This study represents the first time PA prescription has been described across all pharmaceutical families in a large population of companion animals, encompassing PAs authorised for both veterinary and human-only use. These data form a baseline against which future studies could be compared, and provides some useful tools for understanding PA comparative efficacy and risks when prescribed in the varied setting of clinical practice.

Use of vaccines and factors associated with their uptake variability in dogs, cats and rabbits attending a large sentinel network of veterinary practices across Great Britain.

Vaccination remains a mainstay of companion animal population health. However, how vaccine use at a population level complies with existing guidelines is unknown. Here we use electronic health records to describe vaccination in dogs, cats and rabbits attending a large sentinel network of UK veterinary practices. In total, 77.9% (95% CI: 77.6-78.1) of animals had recorded vaccinations. The percentage of animals with recorded vaccinations was higher in dogs, neutered animals, in insured dogs and cats and in purebred dogs. Vaccination rates varied in different regions of Great Britain in all species. Dogs and cats belonging to owners living in less deprived areas of England and Scotland were more likely to be recorded as vaccinated. In the vaccinated population, cats received more core vaccines per year of life (0.86) than dogs (0.75), with feline leukaemia vaccines almost as frequent as core vaccines. In dogs, leptospira vaccines were more frequent than core vaccines. This descriptive study suggests a substantial proportion of animals are not benefiting from vaccine protection. For the first time, we identify potential factors associated with variations in recorded vaccination frequency, providing a critical baseline against which to monitor future changes in companion animal vaccination and evidence to inform future targeted health interventions.

Patterns of antimicrobial agent prescription in a sentinel population of canine and feline veterinary practices in the United Kingdom.

Antimicrobial resistance is an increasingly important global health threat and the use of antimicrobial agents is a key risk factor in its development. This study describes antimicrobial agent prescription (AAP) patterns over a 2year period using electronic health records (EHRs) from booked consultations in a network of 457 sentinel veterinary premises in the United Kingdom. A semi-automated classification methodology was used to map practitioner defined product codes in 918,333 EHRs from 413,870 dogs and 352,730 EHRs from 200,541 cats, including 289,789 AAPs. AAP as a proportion of total booked consultations was more frequent in dogs (18.8%, 95% confidence interval, CI, 18.2-19.4) than cats (17.5%, 95% CI 16.9-18.1). Prescription of topical antimicrobial agents was more frequent in dogs (7.4%, 95% CI 7.2-7.7) than cats (3.2%, 95% CI 3.1-3.3), whilst prescription of systemic antimicrobial agents was more frequent in cats (14.8%, 95% CI 14.2-15.4) than dogs (12.2%, 95% CI 11.7-12.7). A decreasing temporal pattern was identified for prescription of systemic antimicrobial agents in dogs and cats. Premises which prescribed antimicrobial agents frequently for dogs also prescribed frequently for cats. AAP was most frequent during pruritus consultations in dogs and trauma consultations in cats. Clavulanic acid potentiated amoxicillin was the most frequently prescribed antimicrobial agent in dogs (28.6% of prescriptions, 95% CI 27.4-29.8), whereas cefovecin, a third generation cephalosporin, was the most frequently prescribed antimicrobial agent in cats (36.2%, 95% CI 33.9-38.5). This study demonstrated patterns in AAP over time and for different conditions in a population of companion animals in the United Kingdom.

The passive surveillance of ticks using companion animal electronic health records.

Ticks represent a large global reservoir of zoonotic disease. Current surveillance systems can be time and labour intensive. We propose that the passive surveillance of companion animal electronic health records (EHRs) could provide a novel methodology for describing temporal and spatial tick activity. A total of 16 58 857 EHRs were collected over a 2-year period (31 March 2014 and 29 May 2016) from companion animals attending a large sentinel network of 192 veterinary clinics across Great Britain (the Small Animal Veterinary Surveillance Network - SAVSNET). In total, 2180 EHRs were identified where a tick was recorded on an animal. The relative risk of dogs presenting with a tick compared with cats was 0·73 (95% confidence intervals 0·67-0·80). The highest number of tick records were in the south central regions of England. The presence of ticks showed marked seasonality with summer peaks, and a secondary smaller peak in autumn for cats; ticks were still being found throughout most of Great Britain during the winter. This suggests that passive surveillance of companion animal EHRs can describe tick activity temporally and spatially in a large cohort of veterinary clinics across Great Britain. These results and methodology could help inform veterinary and public health messages as well as increase awareness of ticks and tick-borne diseases in the general population.

Quality Assessment of Postmortem Tissue Donors in a University Hospital: Detection and Generation Tissue Indicators.

BACKGROUND: Defining quality assessment and measurement tools in the area of tissue donation should be considered to be one of the most important strategies for developing health centers. The aim of this project was to identify, define, and analyze a set of indicators to assess the most important steps in the tissue donor detection and generation processes. METHODS: A prospective, descriptive, and comparative study of all potential tissue donors (TDs) detected and generated in a university hospital was performed. All deceased patients after cardiocirculatory death were evaluated in 2015 by the transplant coordinators (TCs). We defined as detection indicators: total deaths, percentage of detection and evaluation, percentage of clinical contraindications, tissue donor potentiality (TDP; corneal or multitissue potentiality), and the functional detection time (FDT); and as generation indicators: generation rate (corneal or multitissue generation), family request time, number of interviewed relatives, and TC experience (y). RESULTS: The detection and evaluation rate was 100% (n = 1,235); tissue clinical contraindications were 57%, and TDP was 43% (n = 528; corneal, 80%; multitissue, 20%). The FDT was 24 ± 30 minutes. The generation rate was 53.4% (n = 282): corneal, 57% (n = 241); and multitissue, 40% (n = 41). Family request time was 10 ± 17 minutes, average number of interviewed relatives was 2.2 ± 1.6, and 35% of TCs had experience in the field for >5 years. CONCLUSIONS: Obtaining indicators for quality assessment in the area of tissue donation is useful in predicting the outcome of the TD process as well as promoting the approach of continuous improvement.

Canine babesiosis and tick activity monitored using companion animal electronic health records in the UK.

Recent publications highlighting autochthonous Babesia canis infection in dogs from Essex that have not travelled outside the UK are a powerful reminder of the potential for pathogen emergence in new populations. Here the authors use electronic health data collected from two diagnostic laboratories and a network of 392 veterinary premises to describe canine Babesia cases and levels of Babesia concern from January 2015 to March 2016, and the activity of ticks during December 2015-March 2016. In most areas of the UK, Babesia diagnosis in this population was rare and sporadic. In addition, there was a clear focus of Babesia cases in the affected area in Essex. Until February 2016, analysis of health records indicated only sporadic interest in Babesia largely in animals coming from overseas. Following media coverage in March 2016, there was a spike in owner concern that was geographically dispersed beyond the at-risk area. Tick activity (identified as ticks being removed from animals in veterinary consultations) was consistent but low during the period preceding the infections (<5 ticks/10,000 consultations), but increased in March. This highlights the use of electronic health data to describe rapidly evolving risk and concern that follows the emergence of a pathogen.

European molecular epidemiology and strain diversity of feline calicivirus.

Feline calicivirus (FCV) causes a variable syndrome of upper respiratory tract disease, mouth ulcers and lameness. A convenience-based prospective sample of oropharyngeal swabs (n=426) was obtained from five countries (France, Germany, Greece, Portugal and the UK). The prevalence of FCV by virus isolation was 22.2 per cent. Multivariable analysis found that animals presenting with lymphoplasmacytic gingivitis stomatitis complex were more likely to test positive for FCV infection. Furthermore, vaccinated cats up to 48 months of age were significantly less likely to be infected with FCV than unvaccinated animals of similar ages. Phylogenetic analysis based on consensus sequences for the immunodominant region of the capsid gene from 72 FCV isolates identified 46 strains. Thirteen of the 14 strains with more than one sequence were restricted to individual regions or sites in individual countries; the exception was a strain present in two sites close to each other in France. Four strains were present in more than one household. Five colonies, four of which were rescue shelters, had multiple strains within them. Polymerase sequence suggested possible rare recombination events. These locally, nationally and internationally diverse FCV populations maintain a continuous challenge to the control of FCV infection and disease.

Quercetin and its metabolites inhibit the membrane NADPH oxidase activity in vascular smooth muscle cells from normotensive and spontaneously hypertensive rats.

Quercetin, the most abundant dietary flavonol, exerts antioxidant effects reducing vascular superoxide (O2(-)) and improving endothelial function in animal models of cardiovascular disease. Herein we evaluated the effects of quercetin, and its plasma metabolites, on the nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase activity, the main source of O2(-) in the vessel wall, in vascular smooth muscle cells (VSMCs) from spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY). Quercetin and its metabolites isorhamnetin and kaempferol inhibited the NADPH-stimulated lucigenin-chemiluminescence signal in VSMCs from both strains. The inhibitory effect of quercetin-3-glucuronide increased after prolonged incubation and was inhibited in the presence of the ß-glucuronidase inhibitor saccharolactone. These effects were unrelated to their O2(-) scavenging properties, since they induced only a small inhibition of the rate of pyrogallol autoxidation at high concentrations. All bioflavonoids tested acted as non-competitive inhibitors with respect to NADPH. In conclusion, quercetin and its metabolites inhibit the NADPH oxidase activity in VSMCs reducing O2(-) generation more efficiently than their effect as O2(-) scavengers. The effect of quercetin-3-glucuronide was due to deconjugation and release of free quercetin. The effect is similar in VSMCs from normotensive and hypertensive animals.

Modulation of nitric oxide by flavonoids.

One of the main mechanisms by which dietary flavonoids are thought to influence cardiovascular disease is via protection of the bioactivity of the endothelium-derived nitric oxide (NO). Additionally, flavonoids may also interfere with the signalling cascades of inflammation and prevent overproduction of NO and its deleterious consequences in shock and ischemia-reperfusion injury. In the present paper we review the evidence of the effects of flavonoids on NO. Flavonoids exert complex actions on the synthesis and bioavailability of NO which may result both in enhanced or decreased NO levels: (1) in cell free systems, several flavonoids may scavenge NO via its pro-oxidant properties by increasing superoxide. However, under conditions of oxidative stress, flavonoids may also protect NO from superoxide-driven inactivation. (2) In intact healthy tissues, some flavonoids increase eNOS activity in endothelial cells. Paradoxically this effect involves a pro-oxidant effect which results in Ca(2+)-dependent activation of eNOS. As inhibitors of PI3K, flavonoids may potentially inhibit the PI3K/Akt-dependent activation of eNOS. (3) Under conditions of inflammation and oxidative stress, flavonoids may prevent the inflammatory signalling cascades via inhibition of NFκB and thereby downregulate iNOS. On the other hand, they also prevent the overexpression of ROS generating enzymes, reducing superoxide and peroxynitrite levels, and hence preventing superoxide-induced NO inactivation and eNOS uncoupling. Therefore, the final effect of flavonoids on NO levels will depend on the flavonoid structure and the concentrations used, on the cell type under study and particularly on the presence of inflammatory/oxidative conditions.

Prenatal exposure to hyperoxia modifies the thromboxane prostanoid receptor-mediated response to H2O2 in the ductus arteriosus of the chicken embryo.

O2 tension plays a critical role in the control of prenatal patency and postnatal closure of the ductus arteriosus (DA). We hypothesized that exposure of chicken embryos to hyperoxia alters the morphology and function of DA. Hyperoxia was induced by incubating fertilized eggs at 60% O2 from day 15 to 19 of the 21-d incubation period. DA reactivity (assessed by wire myography), morphometry and mRNA expression of antioxidant enzymes were studied on day 19. Hyperoxic incubation neither affected embryonic growth nor induced signs of DA constriction or changed the mRNA expression of superoxide dismutase and catalase. The contractions induced by O2 (21%), KCl, 4-aminopyridine, phenylephrine, and endothelin-1 and the relaxations induced by acetylcholine (ACh), sodium nitroprusside, isoproterenol, and hydroxyfasudil were similar in DA from embryos incubated under normoxic or hyperoxic conditions. In contrast, hyperoxic incubation impaired the thromboxane prostanoid (TP) receptor-mediated contractions evoked by U46619, 15-E2t-Isoprostane and high concentrations (≥3 µM) of ACh. Exogenous hydrogen peroxide (H2O2) evoked endothelium-dependent contraction in the normoxic DA and endothelium-dependent relaxation in the hyperoxic group. The presence of the TP receptor antagonist SQ 29548 unmasked a relaxant response to H2O2 in the normoxic DA and the cyclooxygenase (COX) inhibitor indomethacin blocked H2O2-induced contraction (in the normoxic group) and relaxation (in the hyperoxic group). Altogether our functional data suggest that, in the chicken DA, exogenous H2O2 induces the release of endothelium-derived COX metabolite(s) with contractile and relaxant properties. Under normal conditions H2O2-induced contraction prevails and relaxation is unmasked after pharmacological or functional (i.e.hyperoxia) TP receptor impairment.

The impact of a specific blend of essential oil components and sodium butyrate in feed on growth performance and Salmonella counts in experimentally challenged broilers.

Essential oils (EO) and short-chain fatty acids have potential antimicrobial activity in broilers. This study aimed to investigate the effect of a specific blend of EO and a combination of this blend of EO with sodium-butyrate on growth performance and Salmonella colonization in broilers. A total of 480 one-day-old male broilers were distributed into 5 treatments (8 pens per treatment and 12 birds per pen) and reared during 42 d in experimental conditions. Dietary treatments consisted of the addition of different doses of EO (0 mg/kg, control; 50 mg/kg, EO50 and 100 mg/kg, EO100) or a combination of EO with 1 g/kg of sodium-butyrate (B; EO50 + B, EOB50 and EO100 + B, EOB100) to a basal diet. All birds were orally infected with 10(8) cfu of Salmonella Enteritidis on d 7 of study. Individual BW and feed intake per pen were measured at arrival and on a weekly basis. The prevalence and enumeration of Salmonella in feces was determined per treatment at 72 h postinfection and on d 23 and 37 of study. At slaughter, cecal content and liver samples from 16 birds per treatment were cultured for Salmonella and cecal pH was measured. No differences were observed on growth performance among treatments. All fecal samples analyzed were positive for Salmonella from d 10 to the end of the rearing period. At slaughter, Salmonella contamination (positive samples) in cecum was lower in birds fed EOB50 compared with the other treatments (P < 0.05), whereas birds fed the control diet showed the highest colonization rates. The pH of the cecal content was not different among treatments. Thus, EO or its combination with sodium-butyrate did not affect growth performance. However, a clear effectiveness of these products was observed in Salmonella control, especially when low doses of EO were combined with sodium-butyrate (EOB50).

PPARß activation restores the high glucose-induced impairment of insulin signalling in endothelial cells.

BACKGROUND AND PURPOSE: PPARß enhances insulin sensitivity in adipocytes and skeletal muscle cells, but its effects on insulin signalling in endothelial cells are not known. We analysed the effects of the PPARß/δ (PPARß) agonists, GW0742 and L165041, on impaired insulin signalling induced by high glucose in HUVECs and aortic and mesenteric arteries from diabetic rats. EXPERIMENTAL APPROACH: Insulin-stimulated NO production, Akt-Ser(473) and eNOS-Ser(1177) phosphorylation, and reactive oxygen species (ROS) production were studied in HUVECs incubated in low- or high-glucose medium. Insulin-stimulated relaxations and protein phosphorylation in vessels from streptozotocin (STZ)-induced diabetic rats were also analysed. KEY RESULTS: HUVECs incubated in high-glucose medium showed a significant reduction in insulin-stimulated production of NO. High glucose also reduced insulin-induced Akt-Ser(473) and eNOS-Ser(1177) phosphorylation, increased IRS-1-Ser(636) and ERK1/2-Thr(183) -Tyr(185) phosphorylation and increased ROS production. The co-incubation with the PPARß agonists GW0742 or L165041 prevented all these effects induced by high glucose. In turn, the effects induced by the agonists were suppressed when HUVEC were also incubated with the PPARß antagonist GSK0660, the pyruvate dehydrogenase kinase (PDK)4 inhibitor dichloroacetate or after knockdown of both PPARß and PDK4 with siRNA. The ERK1/2 inhibitor PD98059, ROS scavenger catalase, inhibitor of complex II thenoyltrifluoroacetone or uncoupler of oxidative phosphorylation, carbonyl cyanide m-chlorophenylhydrazone, also prevented glucose-induced insulin resistance. In STZ diabetic rats, oral GW0742 also improved insulin signalling and the impaired NO-mediated vascular relaxation. CONCLUSION AND IMPLICATIONS: PPARß activation in vitro and in vivo restores the endothelial function, preserving the insulin-Akt-eNOS pathway impaired by high glucose, at least in part, through PDK4 activation.

A multi-analysis approach for space-time and economic evaluation of risks related with livestock diseases: the example of FMD in Peru.

This study presents a multi-disciplinary decision-support tool, which integrates geo-statistics, social network analysis (SNA), spatial-stochastic spread model, economic analysis and mapping/visualization capabilities for the evaluation of the sanitary and socio-economic impact of livestock diseases under diverse epidemiologic scenarios. We illustrate the applicability of this tool using foot-and-mouth disease (FMD) in Peru as an example. The approach consisted on a flexible, multistep process that may be easily adapted based on data availability. The first module (mI) uses a geo-statistical approach for the estimation (if needed) of the distribution and abundance of susceptible population (in the example here, cattle, swine, sheep, goats, and camelids) at farm-level in the region or country of interest (Peru). The second module (mII) applies SNA for evaluating the farm-to-farm contact patterns and for exploring the structure and frequency of between-farm animal movements as a proxy for potential disease introduction or spread. The third module (mIII) integrates mI-II outputs into a spatial-stochastic model that simulates within- and between-farm FMD-transmission. The economic module (mIV) connects outputs from mI-III to provide an estimate of associated direct and indirect costs. A visualization module (mV) is also implemented to graph and map the outputs of module I-IV. After 1000 simulated epidemics, the mean (95% probability interval) number of outbreaks, infected animals, epidemic duration, and direct costs were 37 (1, 1164), 2152 (1, 13, 250), 63 days (0, 442), and US$ 1.2 million (1072, 9.5 million), respectively. Spread of disease was primarily local (<4.5km), but geolocation and type of index farm strongly influenced the extent and spatial patterns of an epidemic. The approach is intended to support decisions in the last phase of the FMD eradication program in Peru, in particular to inform and support the implementation of risk-based surveillance and livestock insurance systems that may help to prevent and control potential FMD virus incursions into Peru.

A new approach for rapidly assessing the risk of Aujeszky's disease reintroduction into a disease-free Spanish territory by analysing the movement of live pigs and potential contacts with wild boar.

Aujeszky's disease (AD) causes significant economic losses in the Spanish pig sector due to import trade restrictions imposed by disease-free countries. Most regions of Spain have achieved 'low AD prevalence' status as a result of an intensive national AD eradication programme involving vaccination and other measures. However, to achieve AD-free status that would eliminate trade restrictions, vaccination must be stopped. For this final stage of eradication, up to date and reliable estimates of the risk of AD reintroduction are essential. Here, we propose an approach based on spatio-temporal scan statistics that the assesses risk of AD reintroduction in a disease-free territory by analysing the two most frequent risk pathways: movement of live domestic pigs and contact with wildlife reservoirs. The approach is illustrated using the case of Navarre, one of the first Spanish regions which plan to stop vaccination. Moreover, direct contacts among pig farms in Navarre were used to evaluate the potential spread of AD in the event of reintroduction. Areas at highest risk of AD reintroduction were in the southern part of the region during the second half of the year through pig movements and in the western and east-central parts of Navarre through contact with wild boars. Northern Navarre, despite having the highest density of pig farms, seems to be at low risk of AD reintroduction. Analysing the network of pig movements within Navarre revealed distinct northern and southern compartments that may be used in preventive compartmentalization strategies to reduce potential risk of AD re-infection in the scenario without vaccination. The approach described here may be extended to other regions and may be useful for guiding risk-based measures that reduce the risk of AD re-infection in a more cost-effective manner. Such analysis in Spain may allow authorities to stop vaccination in the safest possible way.

Social network analysis of Equidae movements and its application to risk-based surveillance and to control of spread of potential Equidae diseases.

Movements of animals and animal products are one of the most important ways of disease introduction and spread between regions and countries. Maybe one of the most complex animal species in terms of diversity of uses, nature and extent of movements are equidae, for which animal movement records are usually not available. The study presented here is the first characterization of a complete and reliable network of equidae movements in Castile and Leon, which is one of the most important equidae production regions of Spain. Social network analysis and space-time cluster analysis were used to describe the contact patterns of the equidae network and to identify the most important premises, areas and time periods for potential disease introduction or spread into the region. The studied network was complex, with very heterogeneous types of premises and diverse nature and extent of the movements compared with other livestock species, which have important implications for prevention and control of equidae diseases. Centrality measures revealed that production and reproduction farms and centres of livestock competition were the most important type of premises in the studied network. Cluster analyses allowed to identify seventeen significant spatio-temporal clusters of premises at high risk of dispatching or receiving equidae, which formed four interconnected compartments. These clusters were mainly located in the north-west region and in the second part of the year. The results of this study may be useful to design risk-based surveillance and control programmes of equidae diseases and increase the speed of detection and control of potential secondary outbreaks in future epidemics. Consequently, these results will help to minimize the great economic and sanitary impact of equidae diseases. The analytical approach used here may be easily extended to characterize the equidae movement patterns in other countries and regions of the world.

Perception of transplant coordinator regarding relatives' attitude toward tissue donation request.

INTRODUCTION: Family denials for tissue donation are higher than denials obtained after organ donation. OBJECTIVES: To find out families' perception toward tissue request as well as its relation with the degree of acceptance or denial. MATERIALS AND METHODS: Following the request for tissue donation (corneas or different tissues), a five closed-question survey was designed and immediately answered by the Transplant Coordinator. The main aim was to find out whether the request determined surprise, disapproval, understanding, knowledge, and immediacy in the decision-making process, as well as its relation with the decision to donate. RESULTS: Of 1751 surveys, 991 were answered and collected from 2009 to 2011, which represents a 56% from the total. Data collected relate to families 510 who donated and 481 who did not (P = NS). The fact of requesting corneas only or different tissues did not influence the answers. Surprise and disapproval variables significantly predicted refusal to donation (P = .000). CONCLUSIONS: Regardless the request type (corneas or multitissue) variables influencing the decision making when approaching families are expression of surprise and disapproval. Neither understanding the application nor existence of a prior approach to donation has any influence whatsoever in the decision-making process.

Different cardiovascular protective effects of quercetin administered orally or intraperitoneally in spontaneously hypertensive rats.

We tested whether the administration procedure of quercetin affects its metabolite profile and antihypertensive activity. Spontaneously hypertensive rats (SHR) were randomly assigned to four experimental treatments: (1) 1 mL of 1% methylcellulose by oral gavage and 2% DMSO i.p. (control group); (2) 10 mg kg⁻¹ quercetin by oral gavage once daily and 2% DMSO i.p.; (3) 10 mg kg⁻¹ quercetin by oral gavage divided in two daily doses (5 + 5 at 12 h intervals) and 2% DMSO i.p.; (4) 1 mL of 1% methylcellulose by oral gavage and 10 mg kg⁻¹ quercetin i.p. injection. Rats were treated daily for 5 weeks. Single dose and two daily doses, in a long-term oral treatment were equally efficient, both restoring the impaired aortic endothelium-dependent vasodilatation and reducing mesenteric contractile response to phenylephrine, systolic blood pressure, heart rate, and heart and kidney hypertrophy. Attenuation of vascular NADPH oxidase-driven O2⁻ production was also found in orally treated rats. Intraperitoneal administration reduced, to lesser extent than oral administration, the increased systolic blood pressure, being without effect to the endothelial dysfunction and vascular oxidative stress. In contrast, greater levels of metabolites were quantified following intraperitoneal compared to oral administration at any time point, except for higher plasma methylated quercetin aglycone in oral as compared to intraperitoneal administration at 2 but not at 8 h. In conclusion, oral quercetin was superior to intraperitoneal administration for the protection from cardiovascular complications in SHR. No differences were found between the oral administration as a single daily dose or divided into two daily doses.

Carotid body function and ventilatory responses in intermittent hypoxia. Evidence for anomalous brainstem integration of arterial chemoreceptor input.

Obstructive sleep apnea is a frequent medical condition consisting in repetitive sleep-related episodes of upper airways obstruction and concurrent events of arterial blood hypoxia. There is a frequent association of cardiovascular diseases and other pathologies to this condition conforming the obstructive sleep apnea syndrome (OSAS). Laboratory models of OSAS consist in animals exposed to repetitive episodes of intermittent hypoxia (IH) which also develop cardiovascular pathologies, mostly hypertension. The overall OSAS pathophysiology appears to be linked to the repetitive hypoxia, which would cause a sensitization of carotid body (CB) chemoreflex and chemoreflex-driven hyperreactivity of the sympathetic nervous system. However, this proposal is uncertain because hyperventilation, reflecting the CB sensitization, and increased plasma CA levels, reflecting sympathetic hyperreactivity, are not constant findings in patients with OSAS and IH animals. Aiming to solve these uncertainties we have studied the entire CB chemoreflex arch in a rat model of IH, including activity of chemoreceptor cells and CB generated afferent activity to brainstem. The efferent activity was measured as ventilation in normoxia, hypoxia, and hypercapnia. Norepinephrine turnover in renal artery sympathetic endings was also assessed. Findings indicate a sensitization of the CB function to hypoxia evidenced by exaggerated chemoreceptor cell and CB afferent activity. Yet, IH rats exhibited marked hypoventilation in all studied conditions and increased turnover of norepinephrine in sympathetic endings. We conclude that IH produces a bias in the integration of the input arising from the CB with a diminished drive of ventilation and an exaggerated activation of brainstem sympathetic neurons.