Experiences and difficulties encountered during a course on veterinary public health with students of different nationalities

Veterinary public health (VPH) issues have received increased attention over the last few years as a result of the rising threat of emerging zoonoses (i.e., those due to globalized trade in animal and animal products and to changes in livestock production systems and the environment). The international dimension of VPH is gradually becoming recognized, and there is a growing need for veterinarians with experience in this field. In order to familiarize (future) veterinarians with the international dimension of VPH, the Department of Public Health and Food Safety of the Faculty of Veterinary Medicine, Utrecht University, has been organizing a course in Veterinary Public Health and Animal Production for over the last 10 years. This course has been intended for Dutch as well as foreign final-year veterinary students and recent veterinary graduates. By bringing together participants from different countries, the course reinforces the international dimension of the issues addressed through the exchange of experiences by the participants themselves. The present article provides information about this course on Veterinary Public Health (VPH): it discusses logistics, didactical approaches, the course program, and the use of information and communication technology (ICT). Special attention is given to the intercultural aspects of higher education, all of which play an important role in the efficient exchange of knowledge between lecturers and students. International courses are an important tool to enable participants to interact in a multicultural environment and address issues that demand international cooperation and a global public health focus.     

A PCR technique for the detection of Jaagsiekte sheep retrovirus in the blood suitable for the screening of ovine pulmonary adenocarcinoma in field conditions

Ovine pulmonary adenocarcinoma (OPA) is a naturally occurring contagious lung neoplasia caused by jaagsiekte sheep retrovirus (JSRV). Although no specific circulating antibodies against the virus can be detected in infected sheep, JSRV proviral DNA sequences can be found in peripheral blood leukocytes (PBLs) in clinically affected and in a proportion of in contact animals. In this study, existing hemi-nested PCR procedure is compared with a new one-step PCR technique that was developed to minimise potential DNA contamination and reduce sample and reagent handling. Different blood preparations were assessed and the best results were achieved on DNA prepared from buffy coat. The sensitivity of this PCR was lower in JSRV infected sheep without lesions of OPA than in clinically affected sheep, which indicate that this PCR may not be not fully appropriate for screening of individual sheep, but rather to provide results at flock level. This PCR is the only currently available blood test for detection of JSRV infected sheep and may be useful in epidemiological studies and in control programmes of OPA.     

Multiple-breed genetic inference using heavy-tailed structural models for heterogeneous residual variances

Multiple-breed genetic models recently have been demonstrated to account for the heterogenous genetic variances that exist between different beef cattle breed groups. We extend these models to allow for residual heteroskedasticity (heterogeneous residual variances), specified as a function of fixed effects (e.g., sex, breed proportion, breed group heterozygosity) and random effects such as contemporary groups (CG). We additionally specify the residual distributions to be either Gaussian or based on heavier-tailed alternatives such as the Student's t or Slash densities. For each of these three residual densities using either homoskedastic (homogeneous variance) or heteroskedastic error specifications, we analyzed 22,717 postweaning gain records from a Nelore-Hereford population based on a Markov chain Monte Carlo animal model implementation. The heteroskedastic Student's t error model (with estimated df = 7.33 +/- 0.48) was clearly the best-fitting model based on a pseudo-Bayes factor criterion. Breed group heterozygosity and, to a lesser extent, calf sex seemed to be marginally important sources of residual heteroskedasticity. Specifically, the residual variance in F1 animals was estimated to be 0.70 +/- 0.16 times that for purebreds, whereas the male residual variance was estimated to be 1.13 +/- 0.09 times that for females. The CG effects were important random sources of residual heteroskedasticity (i.e., the coefficient of variation of CG-specific residual variances was estimated to be 0.72 +/- 0.06). Purebred Nelores were estimated to have a larger genetic variance (124.84 +/- 21.75 kg2) compared with Herefords (40.89 +/- 6.70 kg2) under the heteroskedastic Student's t error model; however, the converse was observed from results based on a homoskedastic Student's t error model (46.24 +/- 10.90 kg2 and 60.11 +/- 8.54 kg2, respectively). These results indicate that allowing for robustness to outliers and accounting for heteroskedasticity of residual variances has potentially important implications for variance component and genetic parameter estimates from data on multiple-breed populations.     

Evaluation of the infectivity of Trichinella papuae and Trichinella zimbabwensis for equatorial freshwater fishes

The discovery of Trichinella species infecting poikilotherm vertebrates has opened new possibilities in the epidemiology of this parasite group. The aim of the present work was to investigate the infectivity of the two non-encapsulated species of Trichinella infecting both mammals and reptiles, Trichinella papuae and Trichinella zimbabwensis, for equatorial freshwater carnivore fishes. To this end, two species of piranhas, four Serrasalmus nattereri and four Serrasalmus rhombeus, were each inoculated per os with the two species of Trichinella larvae. Six days post infection (p.i.), one fish of each species inoculated with one of the two species of Trichinella was sacrificed. The intestines and celomatic cavities were searched for worms using dissection microscopy, and the presence of muscle larvae was evaluated by artificial digestion. The other 4 inoculated fish were sacrificed 60 days p.i. and similarly searched for the presence of worms. No larva or adult worms were detected in any organ or tissue at 6 or 60 days p.i. The lack of infectivity of T. papuae and T. zimbabwensis for fish suggests that the entozoic habitat of this animal does not represent a suitable environment for these two Trichinella species. More importantly, these data indicate that freshwater fishes, one of the food resources for crocodiles, caimans and alligators, are unlikely to play a role in the epidemiology of the known species of the genus Trichinella.     

Morphological, histological, and ultrastructural studies of the ovary of the cattle-tick Boophilus microplus (Canestrini, 1887) (Acari: Ixodidae)

This study presents the morphology of the ovary, as well as the dynamics of the vitellogenesis process in oocytes of the cattle-tick Boophilus microplus. The ovary of these individuals is of the panoistic type; therefore, it lacks nurse cells. This organ consists of a single tubular structure, continuous, and composed of a lumen delimitated by a wall of small epithelial cells with rounded nuclei. In this tick species, the oocytes were classified into six stages varying from I to VI and according to: cytoplasm appearance and presence of the germ vesicle, yolk granules, and chorion. Oocytes of various sizes and at different developmental stages remain attached to the ovary through a cellular pedicel until completing stage V. Afterwards, they are liberated into the lumen and from there to the exterior. Some oocytes (classified as type VI) showed an atypical appearance indicating that some of the cellular components would be undergoing a degenerative process and/or reabsorption.     

Molecular Mapping of the Stb4 Gene for Resistance to Septoria tritici Blotch in Wheat

ABSTRACT Breeding wheat for resistance is the most effective means to control Septoria tritici blotch (STB), caused by the ascomycete Mycosphaerella graminicola (anamorph Septoria tritici). At least eight genes that confer resistance to STB in wheat have been identified. Among them, the Stb4 locus from the wheat cv. Tadinia showed resistance to M. graminicola at both seedling and adult-plant stages. However, no attempt has been made to map the Stb4 locus in the wheat genome. A mapping population of 77 F10 recombinant-inbred lines (RILs) derived from a three-way cross between the resistant cv. Tadinia and the susceptible parent (Yecora Rojo x UC554) was evaluated for disease resistance and molecular mapping. The RILs were tested with Argentina isolate I 89 of M. graminicola for one greenhouse season in Brazil during 1999, with an isolate from Brazil (IPBr1) for one field season in Piracicaba (Brazil) during 2000, and with Indiana tester isolate IN95-Lafayette-1196-WW-1-4 in the greenhouse during 2000 and 2001. The ratio of resistant:susceptible RILs was 1:1 in all three tests, confirming the single-gene model for control of resistance to STB in Tadinia. However, the patterns of resistance and susceptibility were different between the Indiana isolate and those from South America. For example, the ratio of RILs resistant to both the Indiana and Argentina isolates, resistant to one but susceptible to the other, and susceptible to both isolates was approximately 1:1:1:1, indicating that Tadinia may contain at least two genes for resistance to STB. A similar pattern was observed between the Indiana and Brazil isolates. The gene identified with the Indiana tester isolate was assumed to be the same as Stb4, whereas that revealed by the South American isolates may be new. Bulked-segregant analysis was used to identify amplified fragment length polymorphism (AFLP) and microsatellite markers linked to the presumed Stb4 gene. The AFLP marker EcoRI-ACTG/MseI-CAAA5 and microsatellite Xgwm111 were closely linked to the Stb4 locus in coupling at distances of 2.1 and 0.7 centimorgans (cM), respectively. A flanking marker, AFLP EAGG/ M-CAT10, was 4 cM from Stb4. The Stb4 gene was in a potential supercluster of resistance genes near the centromere on the short arm of wheat chromosome 7D that also contained Stb5 plus five previously identified genes for resistance to Russian wheat aphid. The microsatellite marker Xgwm111 identified in this study may be useful for facilitating the transfer of Stb4 into improved cultivars of wheat.     

Characterization of biological activities of feline eosinophil granule proteins

OBJECTIVE:To characterize eosinophil granule-derived proteins in cats. SAMPLE POPULATION: Eosinophils collected via peritoneal lavage from 2 cats. PROCEDURE: The cats were infested orally with Toxocara canis eggs and subsequently challenge-exposed with T. canis antigen injected IP to induce peritoneal eosinophilia; eosinophils were collected via peritoneal lavage. Eosinophil granule proteins were acid-extracted, separated by gel-filtration chromatography, and examined for their peroxidase, ribonuclease, and bactericidal activities; the N-terminal sequence of some of these proteins was determined and compared with homologue proteins from other species. RESULTS: 3 protein peaks were separated in the chromatogram. The first peak had both peroxidase and bactericidal activities. The second peak had ribonuclease and bactericidal activities, and the N-terminal sequence of the major protein was homologous with that of proteins of the ribonuclease A superfamily, including eosinophil ribonucleases from humans and other animal species. The third protein peak had bactericidal activity, and the N-terminal sequence of the major protein was homologous with that of human and murine major basic proteins. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that feline eosinophil granules contain major basic protein and eosinophil-associated ribonuclease and the granule proteins have peroxidase, ribonuclease, and bactericidal activities. In cats, characterization of eosinophil granule proteins may be useful in elucidation of the mechanism of tissue damage in eosinophil-associated diseases and development of new treatment options for those diseases. In addition, the identification of conserved structure and function of eosinophil granule proteins in cats is relevant from an evolutionary viewpoint.     

Changes in chymotrypsin hydrolysis of beta-lactoglobulin A induced by high hydrostatic pressure

Beta-lactoglobulin (beta-Lg) was subjected to high pressures up to 400 MPa and proteolysis with chymotrypsin. The hydrolysates were analyzed by SDS-PAGE and RP-HPLC, and the fragments obtained were identified by ESI-MS/MS. The results obtained showed that beta-Lg was hydrolyzed by chymotrypsin in a "progressive proteolysis" manner at either atmospheric or high pressure. Proteolysis during or after high-pressure treatment showed longer and more hydrophobic peptides than proteolysis at atmospheric pressure. Chymotrypsin showed a behavior similar to that of trypsin, with some differences, probably related to the orientation of the target residues specific for each enzyme. The similarities between proteolytic fragments produced by the two enzymes support that proteolysis enhancement under high pressure depends on the substrate changes rather than the enzyme. High pressure seemed to accelerate the first steps of proteolysis, probably through dimer dissociation, while leaving portions of the molecule more resistant to the enzyme.     

Ontogenic profiling of glucosinolates, flavonoids, and other secondary metabolites in Eruca sativa (salad rocket), Diplotaxis erucoides (wall rocket), Diplotaxis tenuifolia (wild rocket), and Bunias orientalis (Turkish rocket)

As an influence of the Mediterranean diet, rocket species such as Eruca sativa L., Diplotaxis species, and Bunias orientalis L. are eaten all over the world at different ontogenic stages in salads and soups. They are all species within the plant order Capparales (glucosinolate-containing species), and all are from the family Brassicaceae. Predominantly, the leaves of these species are eaten raw or cooked, although Eruca flowers are also consumed. There is considerable potential with raw plant material for a higher exposure to bioactive phytochemicals such as glucosinolates, their hydrolysis products, and also phenolics, flavonoids, and vitamins such as vitamin C. These compounds are susceptible to ontogenic variation, and the few published studies that have addressed this topic have been inconsistent. Thus, an ontogenic study was performed and all samples were analyzed using a previously developed robust liquid chromatography/mass spectrometry method for the identification and quantification of the major phytochemicals in all tissues of the rocket species. Seeds and roots of both Eruca and Diplotaxis contained predominantly 4-methylthiobutylglucosinolate. Leaves of Eruca and Diplotaxis contained high amounts of 4-mercaptobutylglucosinolate with lower levels of 4-methylthiobutlyglucosinolate and 4-methylsulfinylbutylglucosinolate. Flowers of Eruca and Diplotaxiscontained predominantly 4-methylsulfinylbutyl-glucosinolate. In addition, roots of both Diplotaxisspecies contained 4-hydroxybenzylglucosinolate but 4-hydroxybenzylglucosinolate was absent from roots of Eruca. Seeds and seedlings of all Eruca contained N-heterocyclic compounds but no sinapine, whereas Diplotaxis contained sinapine but not the N-heterocycles. In all tissues of B. orientalis, 4-hydroxybenzylglucosinolate and 4-methylsulfinyl-3-butenylglucosinolate were predominant. All rocket tissues, except roots, contained significant levels of polyglycosylated flavonoids, with/without hydroxycinnamoyl acylation. The core aglycones were kaempferol, quercetin, and isorhamnetin. The exception was B. orientalis, which had a negligible seed flavonoid content as compared with the other species. Anthocyanins were only detected in Eruca flowers and consisted of a complex pattern of at least 16 different anthocyanins.     

Putative role of pith cell wall phenylpropanoids in Sesamia nonagrioides (Lepidoptera: Noctuidae) resistance

The stem borer Sesamia nonagrioides (Lefèbvre) is the most important insect pest that attacks maize, Zea mays L., in northwestern Spain. Host plant resistance to this borer was investigated in relation to the cell wall phenylpropanoids content in the pith. Eight inbred lines that differ in resistance were analyzed. Three major simple phenolic acids, p-coumaric, trans-ferulic, and cis-ferulic acids, and three isomers of diferulic acid, 8-5', 8-O-4', and 8-5'b (benzofuran form), were identified. The amount of all these compounds was correlated with the resistance level in the genotypes, with the resistant inbreds having the highest concentrations. The role of these compounds in cell wall fortification and lignification is well-documented, suggesting their possible intervention in S. nonagrioides resistance. Future studies that focus on these compounds could be useful to enhance S. nonagroides resistance.