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Reliability of pressure algometry as an outcome measure in equine research and therapy needs to be studied. The aim of the present study was to establish interexaminer and intraexaminer reliability of pressure algometry in Icelandic horses and to determine reference mechanical nociceptive threshold (MNT) values for that particular breed. Another aim was to create cutoff values, for clinical monitoring of asymmetry in musculoskeletal sensitivity in the Icelandic horse. Nine clinically sound Icelandic horses were tested with a pressure algometer on 11 anatomic landmarks on the neck, back, and croup, each by two examiners. Three weeks later, the procedure was repeated. Interexaminer reliability was good (intraclass correlation [ICC] = 0.64; P

Oral iron therapy can increase the abundance of bacterial pathogens, e.g., Salmonella spp., in the large intestine of African children. Carvacrol is a natural compound with antimicrobial activity against various intestinal bacterial pathogens, among which is the highly prevalent Salmonella enterica serovar Typhimurium. This study aimed to explore a presumed interaction between carvacrol and bacterial iron handling and to assess the potential of carvacrol in preventing the increase of bacterial pathogenicity during high iron availability. S . Typhimurium was cultured with increasing concentrations of iron and carvacrol to study the effects of these combined interventions on growth, adhesion to intestinal epithelial cells, and iron uptake/influx in both bacterial and epithelial cells. In addition, the ability of carvacrol to remove iron from the high-affinity ligand transferrin and an Fe-dye complex was examined. Carvacrol retarded growth of S . Typhimurium at all iron conditions. Furthermore, iron-induced epithelial adhesion was effectively reduced by carvacrol at high iron concentrations. The reduction of growth and virulence by carvacrol was not paralleled by a change in iron uptake or influx into S . Typhimurium. In contrast, bioavailability of iron for epithelial cells was moderately decreased under these conditions. Further, carvacrol was shown to lack the properties of an iron binding molecule; however, it was able to weaken iron-ligand interactions by which it may possibly interfere with bacterial virulence. In conclusion, our in vitro data suggest that carvacrol has the potential to serve as a novel dietary supplement to prevent pathogenic overgrowth and colonization in the large intestine during oral iron therapy.

Federal University of Par?. Laboratory of Wild Animal Biology and Medicine. Bel?m, PA, Brazil / Federal University of Par?. Animal Science Post -graduation Program. Bel?m, PA, Brazil. Federal University of Par?. Laboratory of Wild Animal Biology and Medicine. Bel?m, PA, Brazil / Federal University of Par?. Animal Science Post -graduation Program. Bel?m, PA, Brazil. Federal University of Par?. Laboratory of Wild Animal Biology and Medicine. Bel?m, PA, Brazil / Federal University of Par?. Animal Science Post -graduation Program. Bel?m, PA, Brazil. Federal University of Par?. Biological Sciences Institute. Oxidative Stress Research Laboratory. Bel?m, PA, Brazil. Federal University of Par?. Biological Sciences Institute. Laboratory of In Vitro Fertilization. Bel?m, PA, Brazil. Federal University of Par?. Biological Sciences Institute. Laboratory of Cell Ultrastructure. Bel?m, PA, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Biokapital. Hamar, Norway. Utrecht University. Faculty of Veterinary Medicine. Department of Pathobiology. Utrecht, Netherlands. Institute for Breeding Rare and Endangered African Mammals. Edinburgh, UK. Federal University of Par?. Laboratory of Wild Animal Biology and Medicine. Bel?m, PA, Brazil / Federal University of Par?. Animal Science Post -graduation Program. Bel?m, PA, Brazil. Federal University of Par?. Laboratory of Wild Animal Biology and Medicine. Bel?m, PA, Brazil / Federal University of Par?. Animal Science Post -graduation Program. Bel?m, PA, Brazil. Utrecht University. Faculty of Veterinary Medicine. Department of Pathobiology. Utrecht, Netherlands. Abstract Ovarian fragments were exposed to 0.5 M sucrose and 1 M ethylene glycol (freezing solution; FS) with or without selenium or Trolox. Histological and ultrastructural analyses showed that the percentages of normal follicles in control tissue and in tissue after exposure to FS+50 ?M Trolox were similar. Trolox prevented endoplasmic reticulum (ER)-related vacuolization, which is commonly observed in oocytes and stromal tissue after exposure to FS. From the evaluated stress markers, superoxide dismutase 1 (SOD1) was up-regulated in ovarian tissue exposed to FS+10 ng/ml selenium. Ovarian fragments were subsequently frozenthawed in the presence of FS with or without 50 ?M Trolox, followed by in vitro culture (IVC). Antioxidant capacity in ovarian fragments decreased after freeze-thawing in Troloxfree FS compared with FS+50 ?M Trolox. Although freezing itself minimized the percentage of viable follicles in each solution, Trolox supplementation resulted in higher rates of viable follicles (67 %), even after IVC (61 %). Furthermore, stress markers SOD1 and ERp29 were up-regulated in ovarian tissue frozen-thawed in Trolox-free medium. Relative mRNA expression of growth factors markers was evaluated after freeze-thawing followed by IVC. BMP4, BMP5, CTGF, GDF9 and KL were down-regulated independently of the presence of Trolox in FS but down-regulation was less pronounced in the presence of Trolox. Thus, medium supplementation with 50 ?M Trolox prevents ER stress and, consequently, protects ovarian tissue from ER-derived cytoplasmic vacuolization. ERp29 but not ERp60, appears to be a key marker linking stress caused by freezing-thawing and cell vacuolization.

Zearalenone (ZEN) is a mycotoxin that can be a contaminant of food and feed commodities. ZEN acts as a xenoestrogen and is considered an endocrine disruptor. Since estrogens influence oogenesis during fetal growth, the effect of ZEN on oocytes was investigated in the F1-generation. Pregnant and lactating pigs were exposed to feed naturally contaminated with ZEN (200, 500 and 1000μg/kg feed). Ovaries of F1-animals were examined for follicle development, expression of estrogen converting enzymes and estrogen receptors, and oocyte quality. In F1-newborns, ZEN did not affect follicle dynamics, but follicle integrity decreased with increasing ZEN concentrations. Expression of estrogen receptor beta mRNA increased following ZEN exposure, whereas expression of genes coding for estrogen converting enzymes remained unchanged. In F1-prepubertal gilts, follicular atresia and oocyte maturation with subsequent embryo development remained unchanged. In conclusion, ZEN reduced the quantity of healthy follicles, which may lead to premature oocyte depletion in adulthood. © 2012 Elsevier Inc.

Students in many teacher education programmes experience confusion about portfolios. This study investigated whether using an analogy in the introduction of a portfolio helps teacher education students understand both the purpose of a portfolio and how to compile it. It was also investigated whether these students' understanding correlates with their appreciation of portfolios. In the analogy, portfolios were compared with job application letters, curricula vitae and the references that are common in selection procedures for new employees. This analogy was helpful in promoting students' understanding. Their understanding correlated with their appreciation of this instrument. © 2007 Elsevier Ltd. All rights reserved.

One of the most important factors explaining the distribution and behaviour of coastal marine mammals are tides. Tidal forces drive a large number of primary and secondary processes, such as changes in water depth, salinity, temperature, current velocity and direction. Unravelling which tidal process is the most influential for a certain species is often challenging, due to a lack of observations of all tide related covariates, strong correlation between them, and the elusive nature of most marine organisms which often hampers their detection. In the Marsdiep area, a tidal inlet between the North Sea and the Dutch Wadden Sea, the presence of harbour porpoises ( Phocoena phocoena ) was studied as a function of tide related covariates. Observations were carried out in early spring from a ferry crossing the inlet on a half hourly basis. Environmental and sightings data were collected by one observer, while an on-board Acoustic Doppler Current Profiler (ADCP) and temperature sensor continuously recorded current velocity profiles and temperature, respectively. Sea surface temperature and salinity were measured at a nearby jetty. Sightings ( n = 134) were linked to tidal elevation, geographical position, local depth-averaged current velocity, water temperature (with and without trend correction) and salinity. Variation in sighting rate was best described by salinity, with highest sighting rate at high levels of salinity (> 30 g kg − 1 ), indicating that porpoises enter the area in bodies of (more saline) North Sea water. Second best variable was time of day, with the highest sighting rate early morning, and decreasing during the day. However, surveys in the morning happened to coincide more often with high water and hence, the apparent time of day effect could be due to collinearity. Most porpoises were present in the northern part of the Marsdiep, particularly during high tide. Tide dependent sighting rates confirmed that porpoises reside in the North Sea, and enter the western Wadden Sea during the flood and leave during ebb. This tidal influx is most likely related to prey availability, which corresponds to other recent studies in this area showing higher fish abundance during high tide. Documenting information on tide related patterns could be used in practice, when e.g. planning anthropogenic activities or assessing critical habitats for this species.

Embryonic stem cell (ESC) cultures display a heterogeneous gene expression profile, ranging from a pristine naïve pluripotent state to a primed epiblast state. Addition of inhibitors of GSK3β and MEK (so-called 2i conditions) pushes ESC cultures toward a more homogeneous naïve pluripotent state, but the molecular underpinnings of this naïve transition are not completely understood. Here, we demonstrate that DAZL, an RNA-binding protein known to play a key role in germ-cell development, marks a subpopulation of ESCs that is actively transitioning toward naïve pluripotency. Moreover, DAZL plays an essential role in the active reprogramming of cytosine methylation. We demonstrate that DAZL associates with mRNA of Tet1, a catalyst of 5-hydroxylation of methyl-cytosine, and enhances Tet1 mRNA translation. Overexpression of DAZL in heterogeneous ESC cultures results in elevated TET1 protein levels as well as increased global hydroxymethylation. Conversely, null mutation of Dazl severely stunts 2i-mediated TET1 induction and hydroxymethylation. Our results provide insight into the regulation of the acquisition of naïve pluripotency and demonstrate that DAZL enhances TET1-mediated cytosine hydroxymethylation in ESCs that are actively reprogramming to a pluripotent ground state. Synopsis This study reports that by enhancing Tet1 mRNA translation, the RNA-binding protein DAZL regulates TET1-mediated cytosine hydroxymethylation in ESCs during active reprogramming to a pluripotent ground state. DAZL marks ESCs actively transitioning to a naïve pluripotent state. DAZL enhances Tet1 mRNA translation. DAZL overexpression results in increased hydroxymethylation. This study reports that by enhancing Tet1 mRNA translation, the RNA-binding protein DAZL regulates TET1-mediated cytosine hydroxymethylation in ESCs during active reprogramming to a pluripotent ground state.

Many veterinary curricula use seminars, interactive educational group formats in which some 25 students discuss questions and issues relating to course themes. To get indications on how to optimize the seminar learning process for students, we aimed to investigate relationships between factors that seem to be important for the seminar learning process, and to determine how these seminar factors account for differences in students' achievement scores. A 57-item seminar evaluation (USEME) questionnaire was administered to students right after they attended a seminar. In total, 80 seminars distributed over years 1, 2, and 3 of an undergraduate veterinary medicine curriculum were sampled and 988 questionnaires were handed in. Principal factor analysis (PFA) was conducted on 410 questionnaires to examine which items could be grouped together as indicators of the same factor, and to determine correlations between the derived factors. Multilevel regression analysis was performed to explore the effects of these seminar factors and students' prior achievement scores on students' achievement scores. Within the questionnaire, four factors were identified that influence the seminar learning process: teacher performance, seminar content, student preparation, and opportunities for interaction within seminars. Strong correlations were found between teacher performance, seminar content, and group interaction. Prior achievement scores and, to a much lesser extent, the seminar factor group interaction appeared to account for differences in students' achievement scores. The factors resulting from the present study and their relation to the method of assessment should be examined further, for example, in an experimental setup.

To investigate the relationship between in vitro and in vivo methane (CH4) production measured simultaneously using the same rumen-fistulated cows in both experiments, four dietary treatments based on concentrate that accounted for 400g/kg of the mixed diet DM, were formulated to contain starch varying in rate of fermentation (slowly (S) vs. rapidly (R): native vs. gelatinized maize grain) and level of inclusion (low (L) vs. high (H): 270 vs. 530g/kg of concentrate DM). Sixteen rumen-fistulated lactating dairy cows were used in a complete randomized block design with these treatments replicated in four periods of 17d each. In experiment 1, after 12d of adaptation, the cows were housed in respiration chambers for 5d to measure CH4 production. In experiment 2, in each period in vitro gas and CH4 production were measured (in duplicate per period) for mixed diet samples from the same diet as fed to the donor cows using rumen inocula adapted to the respective diets for an average of 16d. In addition, samples of two concentrate ingredients, viz. grass silage and beet pulp, were incubated with four different inocula obtained from individual donor cows. Gas production (GP) was measured using automated GP system with CH4 measured at distinct time points. In vitro (24-h) CH4 production of mixed diet was lower with R than S (42.9 vs. 49.5ml/g of incubated organic matter (OM); P=0.004), and higher with L than H (49.8 vs. 42.6ml/g of incubated OM; P=0.002). A significant interaction effect between source and level of starch (P=0.015) was also found, indicating the CH4 production of the RH diet decreased in particular. In vivo, an increased rate of starch fermentation resulted in a lower CH4 per unit of estimated rumen-fermentable OM (eRFOM; 55.6 vs. 61.2ml/g of eRFOM; P=0.007), and higher level of starch tended (P=0.089) to reduce CH4 per unit of eRFOM, but dietary starch level and source did not affect CH4 per unit of OM consumed. Across the diets tested, 24-h in vitro CH4 (ml/g of incubated OM) correlated well with in vivo CH4 expressed per unit of eRFOM (R2=0.54; P=0.040), but not when expressed per unit of OM ingested (R2=0.04; P=0.878). For grass silage (the same trend for beet pulp), inocula adapted to R- and H-based diets compared with S- and L-based diets resulted in a lower CH4 production (36.1 vs. 44.8ml/g of incubated OM, R vs. S; and 37.4 vs. 43.4ml/g of incubated OM, H vs. L; P

Clinical Staphylococcus aureus mastitis is difficult to cure. Extended antimicrobial treatment is often advocated as a practical approach to improve cure rates; however, scientific evidence of this hypothesis is lacking. A multi-centered, nonblinded, randomized, positive-controlled clinical trial was conducted in 5 European countries-France, Hungary, Italy, the Netherlands, and the United Kingdom-to study the efficacy of an extended intramammary cefquinome treatment (5 d) compared with a standard intramammary cefquinome treatment (1.5 d) of Staph. aureus clinical mastitis. Least squares means estimates of bacteriological cure during lactation were 34% [standard error (SE). = 9.9%] for the standard treatment group and 27% (SE. = 8.4%) for the extended treatment group. In the final model, extended therapy was not significantly better. The only factor predicting bacteriological cure was pretreatment cow somatic cell count (SCC). Cows with >250,000 cells/mL in milk before treatment were less likely to cure. Least squares means of clinical cure during lactation was 60% (SE. = 19%) for the standard treatment group and 82% (SE. = 12%) for the extended treatment group. In the final model, clinical cure after extended treatment was significantly better. Pretreatment cow udder firmness predicted clinical cure. Firm udders were less likely to cure clinically. Irrespective of treatment regimen, new infection rates with pathogens other than Staph. aureus were higher (42%) after bacteriological cure than after nonbacteriological cure (22%) and cured cows had a significantly lower SCC. In conclusion, independent of the treatment protocol, cows with an SCC

In the classic in situ method, small particles are removed during rinsing and hence their fractional degradation rate cannot be determined. A new approach was developed to estimate the fractional degradation rate of nutrients in small particles. This approach was based on an alternative rinsing method to reduce the particulate matter loss during rinsing and on quantifying the particulate matter loss that occurs during incubation in the rumen itself. To quantify particulate matter loss during incubation, loss of small particles during the in situ incubation was studied using undegradable silica with different particle sizes. Particulate matter loss during incubation was limited to particles smaller than ~40 μm with a mean fractional particulate matter loss rate of 0.035 h−1 (first experiment) and 0.073 h−1 (second experiment) and an undegradable fraction of 0.001 and 0.050, respectively. In the second experiment, the fractional particulate matter loss rate after rinsing in a water bath at 50 strokes per minute (s.p.m.) (0.215 h−1) and the undegradable fraction at 20 s.p.m. (0.461) were significantly larger than that upon incubation in the rumen, whereas the fractional particulate matter loss rate (0.140 and 0.087 h−1, respectively) and the undegradable fraction (0.330 and 0.075, respectively) after rinsing at 30 and 40 s.p.m. did not differ with that upon rumen incubation. This new approach was applied to estimate the in situ fractional degradation rate of insoluble organic matter (OM) and insoluble nitrogen (N) in three different wheat yeast concentrates (WYC). These WYC were characterised by a high fraction of small particles and estimating their fractional degradation rate was not possible using the traditional washing machine rinsing method. The new rinsing method increased the mean non-washout fraction of OM and N in these products from 0.113 and 0.084 (washing machine method) to 0.670 and 0.782, respectively. The mean effective degradation (ED) without correction for particulate matter loss of OM and of N was 0.714 and 0.601, respectively, and significant differences were observed between the WYC products. Applying the correction for particulate matter loss reduced the mean ED of OM to 0.676 (30 s.p.m.) and 0.477 (40 s.p.m.), and reduced the mean ED of N to 0.475 (30 s.p.m.) and 0.328 (40 s.p.m.). These marked reductions in fractional degradation rate upon correction for small particulate matter loss emphasised the pronounced effect of correction for undegraded particulate matter loss on the fractional disappearance rates of OM and N in WYC products.

Sainfoin is a non-bloating temperate forage legume with a moderate-to-high condensed tannin (CT) content. This study investigated whether the diversity of sainfoin accessions in terms of CT structures and contents could be related to rumen in vitro gas and methane (CH4) production and fermentation characteristics. The aim was to identify promising accessions for future investigations. Accessions differed (P < 0·0001) in terms of total gas and CH4> productions. Fermentation kinetics (i.e. parameters describing the shape of the gas production curve and half-time gas production) for CH4 production were influenced by accession (P ≤ 0·038), but not by PEG. Accession, PEG and time affected (P < 0·001) CH4 production, but accession and PEG interaction showed only a tendency (P = 0·08). Increase in CH4 due to PEG addition was not related to CT content. Further analysis of the relationships among multiple traits (nutritional composition, CT structure and CH4 production) using principal component analysis (PCA) based on optimally weighted variables revealed differences among accessions. The first two principal component axes, PC1 (57·6%) and PC2 (18·4%), explained 76·0% of the total variation among accessions. Loading of biplots derived from both PCAs made it possible to establish a relationship between the ratio of prodelphinidin:procyanidin (PD:PC) tannins and CH4 production in some accessions. The PD:PC ratio seems to be an important source of variation that is negatively related to CH4 production. These results suggested that sainfoin accessions collected from across the world exhibited substantial variation in terms of their effects on rumen in vitro CH4 production, revealing some promising accessions for future investigations.

Domestic dogs diverged from grey wolves between 13 000 and 17 000 years ago when food waste from human settlements provided a new niche. Compared to the carnivorous cat, modern-day dogs differ in several digestive and metabolic traits that appear to be more associated with omnivorous such as man, pigs and rats. This has led to the classification of dogs as omnivores, but the origin of these ‘omnivorous’ traits has, hitherto, been left unexplained. We discuss the foraging ecology of wild wolves and calculate the nutrient profiles of fifty diets reported in the literature. Data on the feeding ecology of wolves indicate that wolves are true carnivores consuming a negligible amount of vegetal matter. Wolves can experience prolonged times of famine during low prey availability while, after a successful hunt, the intake of foods and nutrients can be excessive. As a result of a ‘feast and famine’ lifestyle, wolves need to cope with a highly variable nutrient intake requiring an adaptable metabolism, which is still functional in our modern-day dogs. The nutritive characteristics of commercial foods differ in several aspects from the dog's closest free-living ancestor in terms of dietary nutrient profile and this may pose physiological and metabolic challenges. The present study provides new insights into dog nutrition and contributes to the ongoing optimisation of foods for pet dogs.

Summary Background The Global Burden of Disease (GBD) study assesses health losses from diseases, injuries, and risk factors using disability-adjusted life-years, which need a set of disability weights to quantify health levels associated with non-fatal outcomes. The objective of this study was to estimate disability weights for the GBD 2013 study. Methods We analysed data from new web-based surveys of participants aged 18–65 years, completed in four European countries (Hungary, Italy, the Netherlands, and Sweden) between Sept 23, 2013, and Nov 11, 2013, combined with data previously collected in the GBD 2010 disability weights measurement study. Surveys used paired comparison questions for which respondents considered two hypothetical individuals with different health states and specified which person they deemed healthier than the other. These surveys covered 183 health states pertinent to GBD 2013; of these states, 30 were presented with descriptions revised from previous versions and 18 were new to GBD 2013. We analysed paired comparison data using probit regression analysis and rescaled results to disability weight units between 0 (no loss of health) and 1 (loss equivalent to death). We compared results with previous estimates, and an additional analysis examined sensitivity of paired comparison responses to duration of hypothetical health states. Findings The total analysis sample consisted of 30 230 respondents from the GBD 2010 surveys and 30 660 from the new European surveys. For health states common to GBD 2010 and GBD 2013, results were highly correlated overall (Pearson's r 0·992 [95% uncertainty interval 0·989–0·994]). For health state descriptions that were revised for this study, resulting disability weights were substantially different for a subset of these weights, including those related to hearing loss (eg, complete hearing loss: GBD 2010 0·033 [0·020–0·052]; GBD 2013 0·215 [0·144–0·307]) and treated spinal cord lesions (below the neck: GBD 2010 0·047 [0·028–0·072]; GBD 2013 0·296 [0·198–0·414]; neck level: GBD 2010 0·369 [0·243–0·513]; GBD 2013 0·589 [0·415–0·748]). Survey responses to paired comparison questions were insensitive to whether the comparisons were framed in terms of temporary or chronic outcomes (Pearson's r 0·981 [0·973–0·987]). Interpretation This study substantially expands the empirical basis for assessment of non-fatal outcomes in the GBD study. Findings from this study substantiate the notion that disability weights are sensitive to particular details in descriptions of health states, but robust to duration of outcomes. Funding European Centre for Disease Prevention and Control, Bill and Melinda Gates Foundation.

Made available in DSpace on 2014-12-03T13:07:04Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-04-17Bitstream added on 2014-12-03T13:24:33Z : No. of bitstreams: 1 WOS000335177200001.pdf: 750548 bytes, checksum: 075b52b143f984a79a48dec1b9a68ce6 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Background: Descendants from the extinct aurochs (Bos primigenius), taurine (Bos taurus) and zebu cattle (Bos indicus) were domesticated 10,000 years ago in Southwestern and Southern Asia, respectively, and colonized the world undergoing complex events of admixture and selection. Molecular data, in particular genome-wide single nucleotide polymorphism (SNP) markers, can complement historic and archaeological records to elucidate these past events. However, SNP ascertainment in cattle has been optimized for taurine breeds, imposing limitations to the study of diversity in zebu cattle. As amplified fragment length polymorphism (AFLP) markers are discovered and genotyped as the samples are assayed, this type of marker is free of ascertainment bias. In order to obtain unbiased assessments of genetic differentiation and structure in taurine and zebu cattle, we analyzed a dataset of 135 AFLP markers in 1,593 samples from 13 zebu and 58 taurine breeds, representing nine continental areas.Results: We found a geographical pattern of expected heterozygosity in European taurine breeds decreasing with the distance from the domestication centre, arguing against a large-scale introgression from European or African aurochs. Zebu cattle were found to be at least as diverse as taurine cattle. Western African zebu cattle were found to have diverged more from Indian zebu than South American zebu. Model-based clustering and ancestry informative markers analyses suggested that this is due to taurine introgression. Although a large part of South American zebu cattle also descend from taurine cows, we did not detect significant levels of taurine ancestry in these breeds, probably because of systematic backcrossing with zebu bulls. Furthermore, limited zebu introgression was found in Podolian taurine breeds in Italy.Conclusions: The assessment of cattle diversity reported here contributes an unbiased global view to genetic differentiation and structure of taurine and zebu cattle populations, which is essential for an effective conservation of the bovine genetic resources. Univ Estadual Paulista, Fac Ciencias Agr & Vet, BR-14884900 Jaboticabal, SP, Brazil Univ Cattol Sacro Cuore, Inst Zootech, Piacenza, Italy Univ Cattol Sacro Cuore, BioDNA Biodivers & Ancient DNA Res Ctr, Piacenza, Italy Univ Utrecht, Fac Vet Med, Utrecht, Netherlands Univ Giessen, Inst Anim Breeding & Genet, D-35390 Giessen, Germany Univ Estadual Paulista, Fac Med Vet Aracatuba, BR-16050680 Aracatuba, SP, Brazil Univ Estadual Paulista, Fac Ciencias Agr & Vet, BR-14884900 Jaboticabal, SP, Brazil Univ Estadual Paulista, Fac Med Vet Aracatuba, BR-16050680 Aracatuba, SP, Brazil FAPESP: 11/16643-2 FAPESP: 13/12829-0

Salmonella serotyping data, qualitatively described by van Hoek et al. (2012), were used to quantify potential sources of Salmonella in a Dutch pig slaughterhouse. Statistical tests to compare per-day Salmonella prevalence and serotyping data from multiple points in the chain were used to find transmission pathways. A statistical model based on serotyping data was developed to attribute Salmonella on dressed carcasses to the most likely source. Approximately two-third of dressed carcasses carrying Salmonella on the medial surface had been contaminated by house flora. For carcasses carrying Salmonella on the distal surface, transient Salmonella from incoming pigs was a more important source. The relevance of the different sources of Salmonella varied within and between sampling days. Results were compared to those of another modeling approach, in which Salmonella concentration data from the same samples were used (Smid et al., 2012). They mostly agreed. The approach chosen by an individual slaughterhouse depends on the data that are collected.

The five B-subunits (CTB5) of the Vibrio cholerae (cholera) toxin can bind to the intestinal cell surface so the entire AB5 toxin can enter the cell. Simultaneous binding can occur on more than one of the monosialotetrahexosylganglioside (GM1) units present on the cell surface. Such simultaneous binding arising from the toxins multivalency is believed to enhance its affinity. Thus, blocking the initial attachment of the toxin to the cell surface using inhibitors with GM1 subunits has the potential to stop the disease. Previously we showed that tetravalent GM1 molecules were sub-nanomolar inhibitors of CTB5. In this study, we synthesized a pentavalent version and compared the binding and potency of penta- and tetravalent cholera toxin inhibitors, based on the same scaffold, for the first time. The pentavalent geometry did not yield major benefits over the tetravalent species, but it was still a strong inhibitor, and no major steric clashes occurred when binding the toxin. Thus, systems which can adopt more geometries, such as those described here, can be equally potent, and this may possibly be due to their ability to form higher-order structures or simply due to more statistical options for binding. Carbohydrates combating cholera! Simultaneous binding of the five B-subunits of the cholera toxin (CTB5) enhances its affinity and facilitates its cellular entry. Thus, blocking the toxins initial attachment to the cell surface could stop the disease. The binding pattern and potency of tetra- and pentavalent CTB5 inhibitors based on the same GM1 ganglioside scaffold are compared for the first time.

Genetic studies of livestock populations focus on questions of domestication, within- and among-breed diversity, breed history and adaptive variation. In this review, we describe the use of different molecular markers and methods for data analysis used to address these questions. There is a clear trend towards the use of single nucleotide polymorphisms and whole-genome sequence information, the application of Bayesian or Approximate Bayesian analysis and the use of adaptive next to neutral diversity to support decisions on conservation. © 2012 The Authors, Animal Genetics © 2012 Stichting International Foundation for Animal Genetics.

The aim of this study was to examine whether antimicrobial resistance profiles of coagulase-negative Staphylococcus (CNS) species isolated from milk of dairy cows differed between bacterial species, and to compare results obtained by phenotypic and genotypic profiling of resistance to penicillin, oxacillin and macrolide-lincosamide (ML) antibiotics. Of 170 CNS isolates, 83 (48.8%) were phenotypically susceptible to all antimicrobial agents tested in minimum inhibitory concentration (MIC) assays, 40.6% expressed resistance to a single compound or a single class of compounds, and 10.6% to multiple drug classes. Nine percent, 68%, 19%, 4% and 1% of isolates were negative for all resistance genes tested by PCR or positive for one, two, three or four resistance genes, respectively. Phenotypic resistance and detection of resistance genes other than blaZ were relatively rare in Staphylococcus chromogenes, which was the most common CNS species (36% of 170 genotypically identified isolates). In Staphylococcus epidermidis, which was the second most common CNS species (14% of isolates), phenotypic penicillin resistance was significantly more common than in other CNS species. Almost half of the S. epidermidis isolates carried multiple resistance genes and 30% carried the methicillin resistance gene mecA. Survival analysis using MIC values showed significant associations between phenotypic and genotypic resistance profiles. We conclude that CNS species from bovine milk differ significantly in phenotypic and genotypic antimicrobial resistance profiles, which has implications for treatment and management decisions. © 2011 Elsevier B.V.

Y-chromosomal variation in the water buffalo was analysed by sequencing of DBY, ZFY and SRY gene segments. A clear separation of the paternal lineages of the river and swamp types parallels the differences between their maternal lineages and nuclear DNA. Sequence divergence was found to be comparable to the divergence of taurine cattle and zebu, and this divergence predated domestication, confirming that river and swamp buffalo originated from different wild populations. Within a sample of 23 Thai swamp buffaloes, we identified four haplotypes with different geographical distributions, two of which were shared by Thai wild buffaloes. © 2010 Stichting International Foundation for Animal Genetics.

Between January and April 2007, 424 calves under 22 days of age from 108 Dutch dairy herds were sampled to estimate the prevalence of non-normal faeces ('custard-like'-yellowish-coloured with custard consistency or diarrhoea: watery-like faeces) and the shedding of enteropathogens Escherichia coli K99 (E. coli), Coronavirus, Cryptosporidium parvum (C. parvum), Rotavirus and Clostridium perfringens (Cl. perfringens). In addition, information was collected on animal characteristics and herd-management practices. The probability of detecting each one of five enteropathogens given a calf with 'custard-like' faeces or diarrhoea was estimated using Bayes' rule and was based on the predicted probabilities from a multinominal model including each of five enteropathogens as independent variables. In addition, putative risk factors for the presence of each of five enteropathogens were analysed using logistic regression models with random herd effects. Fifty-seven percent of calves had faeces of normal colour (brownish) and consistency (firm), 23.8% (95%CI: 19.8-28.2%) had 'custard-like' faeces and 19.1% (95%CI: 15.5-23.2%) had diarrhoea. E. coli was the least detected enteropathogen (2.6% (95%CI: 1.3-4.6%) of calves, 9% (95%CI: 5-16%) of herds) and Cl. perfringens was most detected (54.0% (95%CI: 49.1-58.8%) of calves, 85% (95%CI: 77-91%) of herds). E. coli and Coronavirus were detected incidentally in only one or two calves per herd, whereas C. parvum and Cl. perfringens were frequently detected in up to four calves per herd. For calves with 'custard-like' faeces, the probability of detecting Rotavirus from a calf in its first week of age was 0.31 whereas for a calf in its second week, there was a 0.66 probability of detecting C. parvum. The probabilities of detecting E. coli, Rotavirus and C. parvum in calves with diarrhoea in their first week of age were 0.10, 0.20 and 0.43, respectively. In calves with diarrhoea between 1 and 2 weeks of age, the probability of detecting enteropathogens was 0.43 for C. parvum. None of the tested enteropathogens were related to 'custard-like' faeces or diarrhoea in the third week of age. Putative risk factors for E. coli, Coronavirus and C. parvum included the presence of peer-calves shedding Coronavirus, C. parvum or Rotavirus, respectively. Additionally, managerial risk factors such as non-optimal hygienic housing (for Coronavirus) and the routine use of antibiotics for diarrhoeic calves (for C. parvum) were found. No animal or managerial factors were associated with shedding of Cl. perfringens. © 2009 Elsevier B.V. All rights reserved.

Background: Two bovine species contribute to the Indonesian livestock, zebu (Bos indicus) and banteng (Bos javanicus), respectively. Although male hybrid offspring of these species is not fertile, Indonesian cattle breeds are supposed to be of mixed species origin. However, this has not been documented and is so far only supported by preliminary molecular analysis. Methods and Findings: Analysis of mitochondrial, Y-chromosomal and microsatellite DNA showed a banteng introgression of 10-16% in Indonesian zebu breeds. East-Javanese Madura and Galekan cattle have higher levels of autosomal banteng introgression (20-30%) and combine a zebu paternal lineage with a predominant (Madura) or even complete (Galekan) maternal banteng origin. Two Madura bulls carried taurine Y-chromosomal haplotypes, presumably of French Limousin origin. In contrast, we did not find evidence for zebu introgression in five populations of the Bali cattle, a domestic form of the banteng. Conclusions: Because of their unique species composition Indonesian cattle represent a valuable genetic resource, which potentially may also be exploited in other tropical regions. © 2009 Mohamad et al.

CD1d-restricted invariant natural killer T cells (NKT cells) have been well characterized in humans and mice, but it is unknown whether they are present in other species. Here we describe the invariant TCR alpha chain and the full length CD1d transcript of pig and horse. Molecular modeling predicts that porcine (po) invariant TCR alpha chain/poCD1d/alpha-GalCer and equine (eq) invariant TCR alpha chain/eqCD1d/alpha-GalCer form complexes that are highly homologous to the human complex. Since a prerequisite for the presence of NKT cells is the expression of CD1d protein, we performed searches for CD1D genes and CD1d transcripts in multiple species. Previously, cattle and guinea pig have been suggested to lack CD1D genes. The CD1D genes of European taurine cattle (Bos taurus) are known to be pseudogenes because of disrupting mutations in the start codon and in the donor splice site of the first intron. Here we show that the same mutations are found in six other ruminants: African buffalo, sheep, bushbuck, bongo, N'Dama cattle, and roe deer. In contrast, intact CD1d transcripts were found in guinea pig, African elephant, horse, rabbit, and pig. Despite the discovery of a highly homologous NKT/CD1d system in pig and horse, our data suggest that functional CD1D and CD1d-restricted NKT cells are not universally present in mammals. (C) 2009 Elsevier Ltd. All rights reserved.

Numerous studies have reported quantitative data on viruses in surface waters generated using different methodologies. In the current study, the impact of the use of either cell culture-based or molecular-based methods in quantitative microbial risk assessment was assessed. Previously and newly generated data on the presence of infectious human enteroviruses (HEV) and enterovirus and parechovirus RNA were used to estimate distributions of virus concentrations in surface waters. Because techniques for the detection of infectious human parechoviruses (HPeV) in surface waters were not available, a ‘Parallelogram Approach’ was used to estimate their concentrations based on the ratio infectious HEV/HEV RNA. The obtained virus concentrations were then used to estimate the probability of exposure for children during recreation in such virus contaminated surface waters. Human enterovirus cell culture/PCR ratios ranged from 2.3 × 10 −3 to 0.28. This broad range of ratios indicates that care should be taken in assuming a fixed ratio for assessing the risk with PCR based virus concentrations. The probabilities of exposure to both enteroviruses and parechoviruses were calculated, using our Parallelogram Approach for the calculation of infectious parechoviruses. For both viruses it was observed that the detection method significantly influenced the probability of exposure. Based on the calculated culture data, PCR data, and the ingestion volume, it was estimated that the mean probabilities of exposure, of recreating children, to surface water containing viruses were 0.087 (infectious enteroviruses), 0.71 (enterovirus particles), 0.28 (parechovirus particles) and 0.025 (calculated infectious parechoviruses) per recreation event. The mean probabilities of exposure of children recreating in surface water from which drinking water is produced to infectious enteroviruses were estimated for nine locations and varied between 1.5 × 10 −4 – 0.09 per recreation event. In this study, the use of the rotavirus dose response relationship as a surrogate was avoided. Instead, the probabilities of exposure were derived as a function of the distributions of the calculated doses. Our ‘Parallelogram Approach’ was used to estimate the unavailable infectious parechovirus concentrations using Monte Carlo simulations, and the exposure assessment carried out showed that virus concentrations present in surface waters could pose a health risk for children and other vulnerable populations.

This study demonstrates for the first time that porcine and mouse sperm incubated in capacitation media supplemented with bicarbonate produce oxysterols. The production is depen- dent on a reactive oxygen species (ROS) signaling pathway that is activated by bicarbonate and can be inhibited or blocked by addition of vitamin E or vitamin A or induced in absence of bicarbonate with pro-oxidants. The oxysterol formation was required to initiate albumin dependent depletion of 30% of the total free sterol and >50% of the formed oxysterols. Incubation of bicarbonate treated sperm with oxysterol-binding proteins (ORP-1 or ORP-2) caused a reduction of >70% of the formed oxysterols in the sperm pellet but no free sterol depletion. Interestingly, both ORP and albumin treatments led to similar signs of sperm capacitation: hyperactivated motility, tyrosin phosphorylation, and aggregation of flotillin in the apical ridge area of the sperm head. However, only albumin incubations led to high in vitro fertilization rates of the oocytes, whereas the ORP-1 and ORP-2 incubations did not. A pretreatment of sperm with vitamin E or A caused reduced in vitro fertilization rates with 47% and 100%, respectively. Artificial depletion of sterols mediated by methyl-beta cyclodextrin bypasses the bicarbonate ROS oxysterol signaling pathway but resulted only in low in vitro fertilization rates and oocyte degeneration. Thus, bicarbonate- induced ROS formation causes at the sperm surface oxysterol formation and a simultaneous activation of reverse sterol transport from the sperm surface, which appears to be required for efficient oocyte fertilization. © 2013 by the Society for the Study of Reproduction, Inc.

Conservation of the intraspecific genetic diversity of livestock species requires protocols that assess between-breed genetic variability and also take into account differences among individuals within breeds. Here, we focus on variation between breeds. Conservation of neutral genetic variation has been seen as promoting, through linkage processes, the retention of useful and potentially useful variation. Using public information on beef cattle breeds, with a total of 165 data sets each relating to a breed comparison of a performance variable, we have tested this paradigm by calculating the correlations between pairwise breed differences in performance and pairwise genetic distances deduced from biochemical and immunological polymorphisms, microsatellites and single-nucleotide polymorphisms. As already observed in floral and faunal biodiversity, significant positive correlations (n=54) were found, but many correlations were non-significant (n=100) or significantly negative (n=11). This implies that maximizing conserved neutral genetic variation with current techniques may conserve breed-level genetic variation in some traits but not in others and supports the view that genetic distance measurements based on neutral genetic variation are not sufficient as a determinant of conservation priority among breeds. © 2011 Blackwell Verlag GmbH.

Both Bos indicus (zebu) and Bos javanicus (banteng) contribute to the Indonesian indigenous livestock, which is supposedly of a mixed species origin, not by direct breeding but by secondary cross-breeding. Here, the analysis of mitochondrial, Y-chromosomal and microsatellite DNA showed banteng introgression of 10-16% in Indonesian zebu breeds with East-Javanese Madura and Galekan cattle having higher levels of autosomal banteng introgression (20-30%) and combine a zebu paternal lineage with a predominant (Madura) or even complete (Galekan) maternal banteng origin. Two Madura bulls carried taurine Y-chromosomal haplotypes, presumably of French Limousin origin. There was no evidence for zebu introgression in five populations of the Bali cattle, a domestic form of the banteng. © 2012 Blackwell Verlag GmbH.

Digital dermatitis (DD) is the most important infectious claw disorder in dairy cattle and herd-based foot bathing with antibacterials, such as 4% formalin, is often used to prevent it. However, there is a lack of long-term studies of the effectiveness of such regimes and in this study the preventive and curative effect of 4% formalin was compared with that of an acidified, ionized copper sulphate solution over a 4-month period on a commercial 120-cow dairy farm. The cows were walked through a split-leg footbath where left claws were treated with an acidified copper solution twice daily for 5. days/week, while right claws were treated with 4% formalin twice daily for 1. day/every second week. Hind claws were scored for the presence and severity of DD in a trimming chute at the start of the study and every 4. weeks throughout the study period. At the start of the study 21/110 cows had ulcerative DD lesions with 10 on the left hind foot, 8 on the right and 3 on both. These lesions, as well as any new lesions which arose during the study, were treated with chlortetracycline spray. During the study 440 observations were made and seven new DD lesions were recorded on left feet (copper treated) and 20 on right feet (formalin treated). Based on survival analysis, the risk of developing a new ulcerative DD lesion on copper-treated (left hind) feet was almost three times less (RR = 0.37, 95% CI 0.16-0.91) than formalin-treated (right hind) feet. Cure rates of DD lesions were not different between copper and formalin. © 2012 Elsevier Ltd.