The Morphology of 16 and 17 Day Old Bovine Blastocysts from Virgin and Repeat Breeder Heifers

The morphology of 16–17 days old embryos from virgin heifers (VH) and repeat breeder heifers (RBH) was compared using light and electron microscopy. In addition some embryos transferred from one heifer category to the other were studied. Embryos from VH were elongated blastocysts and the oval embryonic disc had three germ layers. The ectoderm was stratified and many mitoses were seen. The endoderm lining the blastocoelic cavity consisted of almost squamous cells conjoined by tight junctions. Between the ectoderm and the endoderm the mesoderm had developed and expanded laterally and the coelom had formed. The trophoblastic cells adjacent do the embryonic disc were cylindrical, whereas those more peripheral located were cuboidal. The trophoblastic cells were conjoined by tight junctions and they had numerous long microvilli on their peripheral surface. Except in the embryonic disc region, the endodermal cells had filopodial processes towards the trophoblast. The embryos from RBH varied in appearance. One was similar to those from VH whereas the others were, more or less retarded, without formation of mesoderm. The smaller one consisted ot trophoblastic cells only. The transferred embryos (representing surviving embryos: 2 out of 9 in VH-RBH and 5 of 6 in RBH-VH) had a morphology similar to that of VH blastocytes two though, appeared somewhat retarded. It is suggested that the retarded embryos lack the ability to complete embryonic development and that the uterine environment of RBH is not favourable to sustain normal embryonic development.     

Quasi-non-linear fracture mechanics analysis of splitting failure in simply supported beams loaded perpendicular to grain by dowel joints

A quasi-non-linear fracture mechanics model based on beam on elastic foundation theory is applied for analysis of the splitting failure of dowel joints loaded perpendicular to grain. Simply supported beams symmetrically loaded by two dowels are considered, and the effects of edge distance, dowel spacing, and distance between dowels and supports are accounted for. The foundation modulus used in the beam on elastic foundation model is chosen so that the perpendicular-to-grain tensile strength and fracture energy properties of the wood are correctly represented. This ensures that a conventional stress analysis and failure criterion lead to the same solution as the compliance method of fracture mechanics. A semiempirical efficiency factor is proposed to account for the influence of the total beam depth, which does not enter the beam on elastic foundation model, but the effect of which is evident from tests. It is shown that the so-called Van der Put/Leijten model, which recently has been adopted in Eurocode 5, appears as a special case of the model presented. Tests on simply supported beams with a single dowel joint at midspan are compared with the theoretical predictions. Various edge distances, beam depths, and spans were tested.     

The antimicrobial susceptibility of Staphylococcus species isolated from canine dermatitis

In a retrospective study, 1538 strains of -haemolysin-producing Staphylococcus species isolated from dermatitis in dogs at three veterinary clinical microbiology laboratories in Norway during 1986–87 and 1993–94 were investigated for their antimicrobial susceptibility. None of the strains was resistant to cloxacillin, cephalexin or the quinolones enrofloxacin and ciprofloxacin. More than 96% of the strains were susceptible to trimethoprim-sulphonamide, bacitracin and fucidic acid. Between 67% and 89% of the strains were susceptible to erythromycin, lincosamides, tetracycline, neomycin and chloramphenicol. Only 37.9% of the strains were susceptible to penicillin. The frequency of penicillin resistance increased significantly between the first and second periods, from 46.0% to 58.6%. The frequency of resistance to lincomycin, clindamycin and erythromycin also increased significantly between the first and second periods, from 3.0%, 2.1% and 3.3% to 25.5%, 19.5% and 24.8%, respectively. A moderate increase in resistance to tetracycline was also noted, from 20.4% in the first to 27.6% in the second period. On the other hand, the frequency of resistance to trimethoprim-sulphonamide decreased significantly from 4.1% in the first to 0.9% in the second period. Many different resistance patterns were observed in each period. However, the proportion of multiresistant strains increased from 2.1% in the first to 10.2% in the second period. There was a decrease in resistance to the combination of trimethoprim-sulphonamide and penicillin from the first to the second period. Resistance to the combination of lincosamides and pencillin increased. For the combinations penicillin-tetracycline-lincosamides, pencillin-lincosamides-erythromycin, and pencillin-tetracycline-lincosamides-erythromycin, there was a striking increase in resistance between the first and the second periods.Abbreviations CVL Central Veterinary Laboratory - NCVM Norwegian College of Veterinary Medicine - NVL Norwegian Veterinary Laboratory     

‘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.     

Detection and characterization of phytoplasmas in diseased stone fruits and pear by PCR-RFLP analysis in Turkey

During the late summer-early autumn of 2002, surveys were carried out in Turkey to determine the presence of phytoplasma diseases in fruit trees. Phytoplasmas were detected and characterized by PCR-RFLP analysis and TEM technique in stone fruit and pear trees in the eastern Mediterranean region of the country. Six out of 24 samples, including almond, apricot, peach, pear and plum, gave positive results in PCR assays. RFLP analysis usingSspI andBsaAI enzymes of PCR products obtained with primer pair f01/r01 enabled identification of the phytoplasmas involved in the diseases. Stone fruit trees, including a local apricot variety (‘Sakıt’) and a pear sample, were found to be infected with European stone fruit yellows (ESFY, 16SrX-B) and pear decline (PD, 16SrX-C) phytoplasmas, respectively. This is the first report in Turkey of PD phytoplasma infecting pear and of ESFY phytoplasma infecting almond, apricot, myrobalan plum and peach; ESFY phytoplasma infecting Japanese plum was previously reported. http://www.phytoparasitica.org posting July 21, 2005.     

Characteristics for Practical Use of Attenuated Isolate UA-Fukushima of <Emphasis Type="Italic">Tomato mosaic virus</Emphasis>

L₁₁A-Fukushima (L₁₁A-F) derived from attenuated isolate LuA of Tomato mosaic virus (ToMV) has the highest ability to cross protect against virulent ToMV among LuA and its derivatives and is stably inherited. Growth, yield, fruit quality and symptom attenuation of inoculated tomato plants did not differ significantly between L₁₁A-F and L₁₁A. The infectivity of progeny viruses in tomato infected with LuA-F was less than 4% of that with virulent ToMV. From these results, L₁₁A-F appears to possess the properties necessary for practical use. To manage L₁₁A-F strictly, a PCR-based assay to detect trace contamination of virulent ToMV in L₁₁A-F preparations was established. Received 10 June 2002/ Accepted in revised form 30 October 2002