Quantitative sandwich ELISA for the determination of lupine (Lupinus spp.) in foods

The use of lupine in foods has increased considerably during the past decade, reflected by a corresponding increase in reported lupine-induced allergic incidents. Lupine allergy may arise either by primary sensitization or by clinical cross-reactivity in peanut-allergic persons. Detection of lupine proteins in food has previously been based on the use of patient serum. A novel sandwich enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of lupine in processed foods was developed, using a polyclonal rabbit antilupine capture antibody and a biotinylated conjugate of the same antibody for detection. The antibody was highly specific for lupine, apart from minor cross-reactivities to other legumes. The assay had a detection limit of 1 mug/g and was successfully used to quantify lupine protein in various food matrixes. Recoveries ranged from 60 to 116%, while the intra-and interassay coefficients of variation were <6% and <21%, respectively.     

Establishment and Identification of Cytoplasmic Male Sterility in Brassica napus by Intergeneric Somatic Hybridization

Exploitation of novel cytoplasmic male sterility(CMS)is a main approach for widening the cytoplasmic genetic background of hybrid oilseed rape and avoiding epidemic risk in oilseed rape production.In this study,symmetric somatic hybrids between Brassicanapus var.Zhongshuang4 and Sinapis arvensis(Yeyou18)were produced by protoplast fusion.Two of the six established hybrids were male sterile showing trace or no pollen release upon flowering with non-or slightly extended stamens.Using Zhongshuang4 as a recurrent parent to pollinate the male sterile plants,the ratio of male sterile plants increased with the number of backcrosses.As early as in BC3 generation,most of the sterile families had nearly 100%sterile plants.Up to BC4 generation,the male sterility became stable and no fertility segregation was observed.All F1 progenies from tested crosses using restorer and maintainer lines of Polima CMS were 100%sterile,indicating that the established CMS by somatic hybridization is different from Polima CMS.The origin of the cytoplasm and potential use of this hovel CMS in oilseed rape breeding were discussed.Key wotds:Oilseed rape,Protoplast fusion,Cytoplasmic male sterility,Sinapis arvensis     

Uncommon associations in target resistance among French populations of Myzus persicae from oilseed rape crops

Within the framework of a molecular exploration of target resistance in populations of Myzus persicae on oilseed rapes in France, (1) the S431F mutation (coding gene ace2), although previously reckoned to be rare, revealed to be frequent, (2) M918L (phenotypically characterised) and L932F (both on para) were found for the first time in M. persicae, and (3) a linkage was revealed between M918L and S431F. While until recently populations developing on French oilseed rapes were dominated by genotypes possessing pyrethroid target resistance and esterase overproduction, to date a different type of dominating genotype, equipped with carbamate and pyrethroid target resistance, seems to be invading such fields.     

Breed‐specific variation of hematologic and biochemical analytes in healthy adult Bernese Mountain dogs

Background: Hematology and serum biochemistry reference intervals in dogs may be affected by internal factors, such as breed and age, and external factors, such as the environment, diet, and lifestyle. In humans, it is well established that geographic origin and age may have an impact on reference intervals and, therefore, more specific reference intervals are sought for subpopulations. Objective: The objective of this study was to validate and transfer standard laboratory reference intervals for healthy Bernese Mountain dogs and to create new intervals for analytes where the established laboratory reference intervals were rejected. Methods: The procedure was performed using the human Clinical and Laboratory Standards Institute‐approved model modified for veterinary use. Thirty‐two dogs were included in the study using a direct a priori method, as recommended. Results: While 23 of the standard laboratory reference intervals were readily validated, 7 of the analytes (eosinophils, MCHC, alkaline phosphatase [ALP], γ‐glutamyltransferase, total bilirubin, amylase, and cholesterol) required new reference intervals according to the standard. These were calculated using the robust method. In particular, the new reference range for ALP was wide compared with the established laboratory reference interval. No clinical causes were found for differences in the results of these analytes. Conclusion: We found significant differences in 7 hematologic and serum biochemical analytes for which a breed‐specific variation appears to be the most plausible explanation. Breed‐specific reference intervals for Bernese Mountain dogs will help avoid misinterpretation of laboratory results in the diagnostic process.     

Transiliac approach for exposure of lumbosacral intervertebral disk and foramen: technique description

Objective— To describe and evaluate a transiliac approach to the L7–S1 disk and intervertebral foramen in dogs.
Study Design— Cadaver study.
Animals— Fresh canine cadavers (n=10).
Methods— A craniolateral approach was made to each iliac wing of 10 fresh canine mixed breed cadavers. An 18 mm hole was drilled in a standardized position through the iliac wing. The musculature connected to the cranial aspects of the sacral wing was dissected and retracted cranially through this iliac window. Endoscopic exploration of the area was performed.
Results— The foramen and intervertebral disk were clearly observed in all specimens without iatrogenic injury of the L7 nerve branch. Access to the foramen was possible in 16 of 20 specimens without excision of the sacral wing; however, it was always partially excised to observe the intervertebral disk which lies more caudally and ventrally.
Conclusion— Transiliac approach to the lumbosacral joint allows direct exposure of the intervertebral disk and foramen through an iliac window. Endoscopic exploration provided good observation of the intervertebral disk and/or foramen.
Clinical Relevance— Transiliac approach could be used for lateral corpectomy and foraminotomy in dogs with degenerative lumbosacral stenosis caused by ventral or ventrolateral disk protrusion, foramen stenosis, or OCD of the L7–S1 joint. Clinical study will be necessary to evaluate the efficacy of this approach.     

Exudative epidermitis in pigs caused by toxigenic Staphylococcus chromogenes

Staphylococcus chromogenes is closely related to Staphylococcus hyicus, which is recognised as the causative agent of exudative epidermitis (EE) in pigs. S. chromogenes is part of the normal skin flora of pigs, cattle and poultry and has so far been considered non-pathogenic to pigs. A strain of S. chromogenes producing exfoliative toxin type B, ExhB, was identified by the use of a multiplex PCR specific for the exfoliative toxins from S. hyicus. The exfoliative toxin from S. chromogenes reacted in immunoblot analysis with polyclonal and monoclonal antibodies specific to ExhB from S. hyicus and had an apparent molecular weight of 30 kDa. Sequencing the gene encoding the exfoliative toxin from S. chromogenes revealed that the molecular weight of the toxin with the signal peptide and the mature toxin was 30,553 and 26,694 Da, respectively. Comparison of the exhB genes from S. chromogenes strain VA654 and S. hyicus strain 1289D-88 showed differences in seven base pairs of the DNA sequences and in two amino acid residues in the deduced amino acid sequences. Pigs were experimentally inoculated with S. chromogenes strain VA654. By clinical observations and histopathological evaluation of the skin alterations, all pigs revealed development of generalized exudative epidermitis. No toxin producing S. hyicus was isolated from the pigs and all ExhB-positive bacterial isolates were identified as S. chromogenes. This confirmed that the disease-causing agent was the inoculated S. chromogenes strain VA654. The results of this study show that S. chromogenes may cause exudative epidermitis in pigs.     

Selection of Nontarget Testing Organisms for ERA of GM Potato with Increased Resistance to Late Blight

The deliberate release of any genetically modified (GM) organism in the European Union requires an environmental risk assessment (ERA) prior to commercialisation, including impact assessment on nontarget organisms. We report from two expert workshops where a newly developed selection procedure for identification of ecologically relevant testing organisms was applied to the case of a GM potato with increased resistance to late blight, planned for cultivation in southern Scandinavia. Species known to contribute to important ecological functions in the receiving environment were selected in a stepwise procedure, to arrive at a practical number of ecologically relevant species that are likely to be exposed to the transgene and suitable for experimental testing. Four ecological functional categories were identified: herbivory and disease transmission, natural enemies, ecological soil processes and pollination. Among these, relevant nontarget species were identified for herbivores and soil living pathogens, natural enemies and decomposers/beneficial soil organisms. Out of a total of 16 herbivores, 17 soil-living pathogens, 49 natural enemies and 14 decomposers/beneficial soil organisms in the initial lists, 8 herbivores, 10 soil-living pathogens, 15 natural enemies and 11 decomposers/beneficial soil organisms were identified as possible testing organisms, based on ecological criteria. These findings are highly relevant for determining the scope and structure of an ERA of this type of GM potato. The selection procedure could not be completed because of insufficient information about tissue- and developmental stage-specific expression levels of the transgenic products for this particular GM potato. Thus, the case study illustrates some of the difficulties and knowledge gaps that limit the relevance and quality of ERA of GM plants.