Antibody detection-based differential ELISA for NDV-infected or vaccinated chickens versus NDV HN-subunit vaccinated chickens

With the advent of subunit vaccines for microbial diseases it is becoming increasingly important to be able to differentiate naturally infected animals from those vaccinated with the corresponding subunit vaccine. For avian viruses such as Newcastle disease virus (NDV), a whole virus-based ELISA cannot make such a differential diagnosis since in both cases the antisera would react with the whole virus. The nucleocapsid protein (NP) gene of the NDV Hitchner B1 strain was cloned, sequenced and expressed to develop a differential ELISA. The B1 NP had 95.7 and 96.1% amino acid identities with the NP of the d26 and Ulster 2C strains, respectively. The B1 NP expressed in a baculovirus expression vector (recNP) was the expected size and reacted with NDV-specific antibodies (Ab) in Western blots and by radioimmunoprecipitation. The ELISA using recNP-coated wells, tested on serum samples from flocks pretested with a commercial NDV kit gave results corresponding to those of the kit. Furthermore, use of both the renNP-based ELISA and a whole virus ELISA allowed the differentiation of birds vaccinated and a NDV haemagglutinin-neuraminidase (HN) expressing fowlpox virus from birds infected with NDV. This provides the basis for establishing an ELISA that discriminates between the antibody response to a recombinant fowlpox vaccine (expressing NDV HN protein) and that to live and inactivated NDV.     

Development of the simultaneous detection of Ralstonia solanacearum race 3 and Clavibacter michiganensis subsp. sepedonicus in potato tubers by a multiplex real-time PCR assay

Clavibacter michiganensis subsp. sepedonicus and Ralstonia solanacearum (Smith) Yabuuchi et al. race 3 are the causal agents of ring-rot and brown-rot of potato respectively. These diseases represent a serious threat to potato production in temperate climates. Both bacteria are listed as A2 pests in the EPPO region and as zero-tolerance quarantine organisms in the European Union. All the detection tests developed so far were only focused on the detection of a single pathogen while the absence of both bacteria has to be certified in the seed tubers. We have therefore developed a new multiplex real-time PCR assay to simultaneously detect both bacteria in a single assay. Additionally, the reliability of this molecular diagnostic test has been improved by the simultaneous amplification of an internal control, corresponding to a potato gene co-extracted from the sample. The polyvalence and the specificity of each set of bacterial primers and probes were evaluated on more than 90 bacterial strains. The limit of detection of this triplex real-time protocol was similar to those observed with other molecular protocols previously developed for the individual detection of one of these bacteria. A concordance of 100 % was obtained in a blind test mimicking the routine application of the technology. In conclusion, this new protocol represents a straightforward and convenient method potentially adapted to primary screening of potato tubers.     

Tree-Based Methods as an Alternative to Logistic Regression in Revealing Risk Factors of Crib-Biting in Horses

Determining the risk factors might help in designing prevention of crib-biting. Logistic regression is a commonly used statistical method for finding risk factors, but tree-based methods are also getting more popular. An important difference between these two statistical approaches is that logistic regression makes a number of assumptions about the underlying data, whereas tree-based methods do not. Another difference is that logistic regression can be used to derive odds ratios for the significant risk factors, whereas tree-based methods create a tree where the ramifications represent the risk factors. The probability of occurrence is assigned to each end of branch in the tree. Data of horses used for noncompetition purposes were analyzed with three statistical approaches: logistic regression, classification tree, and conditional inference tree methods. By this, we compared the advantages and disadvantages of these statistical methods. No difference was found between the two tree-based methods regarding the structure and prediction accuracy of the trees. Compared to them, logistic regression revealed fewer risk factors, and also the number of the stereotypic horses classified correctly by the model was less. The representation of the tree-based methods is closer to medical reasoning and also high-order interaction of the risk-factors can easily be visualized. Our results suggest that tree-based methods can be a new alternative in revealing risk factors, even if used alone or together with logistic regression.     

Characteristics of prolactin-releasing response to salsolinol in vivo in cattle

The aims of the present study were to clarify the effect of salsolinol (SAL), a dopamine (DA)-derived endogenous compound, on the secretion of prolactin (PRL) in cattle. The experiments were performed from April to June using calves and cows. A single intravenous (i.v.) injection of SAL (5 mg/kg body weight [BW]) or sulpiride (a DA receptor antagonist, 0.1 mg/kg BW) significantly stimulated the release of PRL in male and female calves (P < 0.05), though the response to SAL was smaller than that to sulpiride. The secretory pattern of PRL in response to SAL or sulpiride in female calves resembled that in male calves. A single i.v. injection of SAL or sulpiride significantly stimulated the release of PRL in cows (P < 0.05). There was no significant difference in the PRL-releasing response between the SAL- and sulpiride-injected groups in cows. A single intracerebroventricular injection of SAL (10 mg/head) also significantly stimulated the release of PRL in castrated calves (P < 0.05). These results show that SAL is involved in the regulatory process for the secretion of PRL, not only in male and female calves, but also in cows. The results also suggest that the potency of the PRL-releasing response to SAL differs with the physiological status of cattle.     

Validation of Dual‐energy X‐ray Absorptiometry to Predict Body Composition of Channel Catfish,Ictalurus punctatus

Dual‐energy X‐ray absorptiometry (DXA) provides a noninvasive way to determine lean tissue mass (LTM), fat mass (FM), bone mineral content (BMC), and bone mineral density (BMD) in humans and small mammals. Live channel catfish (n = 74, 78–1200 g) were anesthetized and scanned in both a lateral position and a dorsa‐ventral position. Six individual fish (300–600 g) were scanned five times each to determine precision by the coefficient of variation. Precision was good for LTM (0.75–1.06%) and for BMC and BMD (2–2.6%). Precision for FM was not good (27–34%), which was due to the very low FM (0–1 g) recorded by the DXA. However, using the predicted values, FM precision improved to 5–5.5%. DXA values for LTM, FM, and BMC were significantly different from chemical analysis (P < 0.001). DXA overestimated LTM and underestimated FM and BMC. However, all three compartments were strongly correlated with carcass values (P < 0.0001). Using the prediction equations and the jackknife procedure, predicted values of LTM, FM, and BMC were not significantly different from the carcass values (P > 0.05). DXA may also be a valuable tool for evaluating body condition longitudinally in commercial or in threatened or endangered fish species, where noninvasive procedures would be invaluable.     

Enterotoxigenic Escherichia coli (ETEC) in farm animals.

Animal diseases due to enterotoxigenic Escherichia coli (ETEC) typically appear as severe watery diarrhoea during the first few days of life (also a few days after weaning in pigs). ETEC adhere to the small intestinal microvilli without inducing morphological lesions and produce enterotoxins acting locally on enterocytes. This action results in the hypersecretion (of water and electrolytes) and reduced absorption. Adhesins and toxins are the two prominent virulence attributes of ETEC and the level of knowledge of these factors determines the chances for successful prevention and therapy of the disease. For animal ETEC the most common adhesins are the fimbriae (pili) on the surface: F4(K88), F5(K99), F6(987P), F41, F42, F165, F17 and F18. Enterotoxins (extracellular proteins or peptides) of animal ETEC are classified as heat-labile (LT) and heat-stable (ST) enterotoxins with further subdivisions (LTh-I, LTp-I, LTIIa, LTIIb, STaH, STaP, STb) according to antigenic and biological differences. Fimbriae and LT enterotoxins are made up of large molecular weight proteins which facilitate their utilisation in vaccines and their detection using immunodiagnostic systems. The adhesive fimbriae and enterotoxins of animal ETEC are plasmid determined (except F41 and F17). Virulence gene probes (DNA hybridisation, PCR) are specific and sensitive diagnostic and epidemiologic tools for the detection of ETEC. Based on genetic typing, the ETEC, in spite of limited serogroups, seem to represent a population of E. coli with a diverse genetic background.