Urinary protein level determination as a rapid method for the detection of contamination of slaughtered swine

Reference is made to work undertaken by Stehle (1983) on rapid semiquantitative determination of urinary protein in pigs for slaughter to identify premortal strain and stress, with additional studies being conducted into the suitability of the method. The investigations performed for this study covered urinary protein samples from 59 living and 762 slaughtered pigs. The Biophan E paper strip test was applied to groups of animals, and, when compared to high-accuracy laboratory determination of urinary protein, it provided sufficiently accurate information on the presence of stress-related proteinuria. Its accuracy, however, proved to be insufficient, when individual animals were examined. The number of clearly stressed slaughter pigs was unexpectedly high, although the same animals had been rated clinically inconspicuous prior to slaughter. Brief rest period resulted in significant rise of stress. The Biophan E paper strip test was found to be suitable for instantaneous determination of stress and strain on pigs for slaughter.     

Skeletal muscle sarcoplasmic calcium regulation and sudden death syndrome in chickens

1. Sudden Death Syndrome (SDS) is a disease of well‐developed, predominandy male broiler chickens where death appears to occur because of cardiovascular failure. The role of skeletal muscle sarcoplasmic calcium regulation as a potential cause of SDS has been investigated.

2. Calcium regulation of skeletal muscle sarcoplasmic reticulum was compared between broiler and Leghorn chickens. Calcium regulation matured from the 2nd to the 11 th week and, at any age, broiler chickens showed significandy lower calcium transport rates and transport efficiencies. The mechanism of calcium transport in broiler chickens was more energy‐consuming than that of the Leghorn chickens.

3. Sarcoplasmic calcium regulation is pivotal for muscle metabolism. As in porcine malignant hyperthermia, weaker calcium regulation might lead to hyperactivation of skeletal muscle, followed by elevated lactic acid concentrations and cardiovascular failure.     

‘Plantibodies’: a flexible approach to design resistance against pathogens

Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (V_L) and heavy chain (V_H) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.