Follicullar Development and Embryo Recovery Following 3 versus 8 FSH Injections in Heifers

Ovarian follicular dynamics and embryo yield were studied during 2 different FSH regimens for superovulation of cattle. Twenty heifers were given intramuscular injections of FSH (total of 35 mg NIH) either once daily for 3 days (Group 3×1) or twice daily for 4 days (Group 4×2). At 72 h after the first FSH injection, each animal was injected with 0.75 mg cloprostenol. Inseminations were performed at 12 h and 24 h after the onset of heat. Transrectal ultrasonography was performed on the day of the first FSH injection, the day of cloprostenol injection, the day of insemination and finally on the day of embryo recovery (day 6 or 7 after heat). The numbers of small (2–4 mm), medium (5–9 mm) and large (>10 mm) size follicles were recorded. The total number of corpora lutea, eggs and transferable embryos were recorded on the day of embryo recovery. No differences were found between the 2 groups in either of the parameters studied (p>0.05). It can be concluded that treatment with this FSH preparation once daily for 3 days gives a folliculogenic and superovulatory response similar to a treatment regimen where it is given twice daily for 4 days.     

Effects of epidural xylazine on EEG responses to surgical stimulation during isoflurane anaesthesia in dogs

The effects of epidural administration of 250 μg/kg xylazine on EEG responses to surgical stimulation of 5 different intensities were evaluated during isoflurane anaesthesia for an experimental orthopaedic procedure in dogs. The dogs were assigned randomly to one of 2 treatment groups receiving either xylazine (n = 4) or equal volumes of sterile water (n = 4) (control group) epidurally. Intense surgical stimulation during removal of a bone graft from the dorsoiliac spine of the ileum was associated with a significantly (P = 0.0339) higher increase in EEG alpha/delta ratio after epidural administration of sterile water than after epidural injection of 250 μg/kg of xylazine. In addition, the preincision baseline values for 80% spectral edge frequency were significantly (P = 0.0339) lower in the xylazine group compared to control dogs. Our results suggest that epidural administration of 250 μg/kg of xylazine during orthopaedic procedures in dogs exerts antinociceptive effects which may be in part mediated by a supraspinal effect of xylazine.     

Utilization of free and protein-bound lysine in the Japanese quail

The utilization of dietary lysine for protein synthesis is affected by the digestibility of protein-bound lysine, by its intestinal resorption and by its oxidative catabolism. The approach chosen in this paper enables a comparison of the cumulative effect of these processes on the utilization of free and protein-bound lysine, respectively. The principle of the approach is based on a quantification of the expiration of 14C-labelled carbon dioxide after an oral administration of a diet, which contains L-(U-14C)-lysine either as a free amino acid or bound to yeast proteins. During an adaptation phase cockerels of the Japanese quail received a diet based mainly on ground wheat and wheat gluten. This diet was supplemented either with yeast proteins or with a mixture of L-amino acids which simulates the composition of the yeast proteins. In the main experiment the expiration of labelled carbon dioxide was measured during 240 minutes after the administration of the corresponding labelled diets. Just before treatment the animals were either in the postprandial phase or in a state of slight hunger. The maximum of expiration of labelled carbon dioxide occurred around the 60th minute after administration of the corresponding labelled diets. The cumulative expiration of labelled carbon dioxide, expressed in per cent of the radioactive dose used, amounts to 15.5% and 14.3% for free and protein-bound lysine, respectively. The utilization of both forms of lysine in the Japanese quail is lower than in broilers.     

Sugar beet mash silage as a component of a total-mixed-ration for dairy cows--effects on parameters of digestion and animal performance.

Sugar beet mash silage (BMS) was offered in amounts up to 35% of DM to dairy cows as a component of a total-mixed-ration (TMR). Barley and molasses in the control ration were replaced by BMS half in ration BMS 1/2 and in total in ration BMS 1/1 on the basis of the calculated contents of net energy for lactation. Two trials were carried out. In trial I each ration was tested on parameters of rumen fermentation and digestibility of crude nutrients using 4 Holstein cows with rumen fistula. Chewing activity was tested on 2 Holstein cows for each ration. With the BMS rations the ruminal NH3 concentration was lower and the drop in pH was less than in the control ration. The pattern of volatile fatty acids in the rumen fluid from the BMS groups tended towards more propionic and butyric acid. The feeding of BMS showed no negative impact on chewing and rumination. Energy digestibility raised significantly from 59.8% in the control ration to 72.6% in the BMS 1/1 ration. In trial II the same rations were fed in a change-overdesign to a herd of 24 Holstein cows to test feed intake and animal performance. The results showed no significant effects of BMS rations on DM intake and milk production. The results of both trials indicate that even high amounts of cereals can be replaced by BMS without negative effects on rumen fermentation, milk yield and milk composition with slight drop in fat content. For a better handling of BMS, it is of advantage to include it in a TMR.     

Enzyme arrangement of various tissues in swine. 1. Studies of the effect of 32 hours of fasting, administration of actinomycin D, and fasting in combination with administration of actinomycin D on crude protein level, activities of GOT, GTP, fructose-1,6-diphoaphatase as well as ATPase of liver, kidneys and semitendinous muscle in newborn piglets]

The effects of sufficient milk intake as well as of 32 hours of fasting after birth, administration of actinomycin D (intraperitoneal application of 1 mg/kg five weight), and fasting in combination with actinomycin D on the development of body and liver weights, crude protein levels in homogenate and supernatant of liver, kidneys, and M. semitendinosus as well as on the activities of certain tissue enzymes were analysed with four groups of piglets (n = 4). Fasting, administration of actinomycin, and fasting in combination with actinomycin D resulted in rapid reduction in body and liver weights, while the crude protein levels in those tissues were not affected with significance. GOT and fructose-1,6-diphosphatase activities in supernatant from liver tended to decline under fasting conditions. The ATPase activity in the homogenate of the above tissues did not change in response to differentiated treatment.     

‘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.     

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