Effects of phenylbutazone and indomethacin on the post-operative course following experimental orthopaedic surgery in dogs

Randomized placebo-controlled crossover studies were carried out in dogs to evaluate how two non-steroidal anti-inflammatory drugs (NSAID) might modulate an acute post-traumatic inflammatory reaction. Two "identical" surgical interventions were performed on the forelimbs of each animal with an interval of 28 days, to enable a paired comparison of the inflammatory signs and the wound/bone healing processes. At one operation 8 dogs received 300 mg phenylbutazone twice daily for 8 days starting on the day before surgery, and at the other operation matching placebo tablets were given. In a similar placebo-controlled trial another group of 8 dogs received 5 mg indomethacin twice daily. With phenylbutazone the post-operative swelling was not significantly reduced compared to placebo, but there was less pain and limping. With indomethacin the swelling was somewhat reduced, but there was no consistent difference to placebo in the pain and limping assessments. None of the drugs appeared to distinctly effect the wound or fracture healing, as evaluated by clinical inspection, comparison of radiographs and comparison of bone sections from the sites of surgery. It proved difficult to select an appropriate dosage of indomethacin due to its high potential to induce GI ulceration and bleeding in dogs. In this experimental surgical model with an acute inflammation, neither phenylbutazone nor indomethacin showed impressive anti-inflammatory or analgesic properties. In the same model paracetamol has proved to significantly and more efficiently, reduce both swelling and pain without any noticeable adverse effects, and appears to be a better alternative than the two presently tested NSAID.     

The potential use of plasma glutamate dehydrogenase activity for the evaluation of hepatic disease in the cockatiel (Nymphicus hollandicus)

A reference interval for plasma glutamate dehydrogenase (GD) (E.C. 1.4.1.3) activity, of 0-8 IU/L, was established for the cockatiel (Nymphicus hollandicus). An automated modification of a commercial manual assay was used. This enzyme is considered liver specific in humans and numerous domestic animals, due to its organ distribution. A similar distribution was found in cockatiels in this study. Maximal enzyme activity was recovered from liver and kidney homogenate supernatants. Minimal activity was detected in skeletal muscle preparations. These results suggest a potential use for plasma GD activity in the evaluation of hepatocelluar injury/necrosis in cockatiels.     

‘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.     

Evaluation of a radioimmunoassay for type III procollagen peptide in the dog

Type III procollagen peptide (P-3-P) is a serum marker for hepatic fibrosis in humans. The utility of a commercially available radioimmunoassay for P-3-P was evaluated in the dog. The specificity of the assay was assessed by polyacrylamide gel electrophoresis (PAGE) of canine serum and purified bovine P-3-P, followed by Western immunoblotting with rabbit aniti-P-3-P serum. The sensitivity was assessed by performing the radioimmunoassay on dilutions of sera from 22 dogs. Polyacrylamide gel electrophoresis of purified bovine P-3-P and sera from two dogs suspected of having elevated P-3-P concentrations revealed no homologous bands of staining. Western immunoblotting showed marked cross-reactivity of the high antisera concentrations with several components of the serum proteins, but none corresponding to the purified P-3-P. All tested sera from dogs had minimal competitive binding with radiolabeled P-3-P in the radioimmunoassay. Dilution curves of dog sera did not parallel either the standard curve or the dilution curve of a known test human serum. There were no statistically different P-3-P concentrations in any of the groups of dogs studied. It was concluded that currently available radioimmunoassay kits for the measurement of P-3-P in the human are not applicable in the dog. Seemingly, the structure or metabolism of canine P-3-P may vary significantly from that of the bovine or human, limiting the sensitivity and specificity of this assay in the dog.     

Factors affecting the hatchability of eggs from broiler breeders

1. Four experiments were carried out on eggs from broiler breeding flocks between 26 and 60 weeks of age. The effects of storage and incubation conditions on hatchability were tested.

2. Collecting eggs hourly rather than five hours after lay slightly reduced hatchability (P<0.10). Pre‐storage fumigation of almost un‐contaminated eggs had no effect on hatchability even after storage for 8 d. Storing eggs in unsealed polythene bags did not affect hatchability of eggs stored for 5 or 8 d.

3. Eggs stored for 2 d hatched better when held at 18 °C than at 15 °C (P<0.05). Eggs stored for 8 d at 15 °C hatched better than eggs stored for 8 d at 18 °C (P< 0.01). Best hatchability was in eggs stored in unsealed polythene bags at a room temperature of 15 °C. When older eggs were allowed 30 to 40 min more in the setter for each day of storage, the decline in hatchability was 0.5 to 0.6 percentage units per day in storage as compared with a decline of 1.2 percentage units per day when eggs of different storage times, up to 8 d, were set simultaneously.

4. Those eggs which showed a weight loss during incubation of near average for their relative humidity (RH) treatment tended to hatch better than others except under conditions of very low RH (0.36), when best hatchability was associated with lower than average weight loss.

5. In eggs from a young flock (28 to 44 weeks of age) hatchability of fertile eggs was depressed by 1 percentage unit with an increase in RH of 0.17, and by 1 percentage unit with each decrease of 0.06 in RH from a control RH of 0.53. In eggs from the same flock between 48 and 60 weeks of age hatchability was depressed by 1 percentage unit with each 0.037 increase in RH from 0.44 to 0.70.

6. Eggs from a young flock (34–49 weeks) hatched significantly better when maintained at 0.82 rather than at 0.66 (P<0.05) or 0.95 (P<0.10) RH during the hatching period from 19 to 21 d of incubation. Eggs from an older flock (51–61 weeks) hatched better at 0.82 and at 0.‐92 than at 0.72 RH during the same period, but the differences were not significant.     

The effects of progesterone on oviposition and ovulation in the domestic fowl (Gallus domesticus)

1. Following an injection of 0.5 or 0.1 mg progesterone/kg between 0 and 6 h after ovulation, oviposition of the resulting egg was delayed by 1 to 11 h and occurred 26 to 31 h after injection, depending on the dose. The injection terminated the laying of a sequence of eggs by causing the next ovulation to occur a day late. The delayed ovulation occurred at the time normally expected for the first ovulation of a sequence and became the first of a new sequence.

2. Following an injection of 0.5 or (H mg progesterone/kg between 6 and 15 h after ovulation, oviposition of the resulting egg was generally delayed by between 15 and 28 h and occurred at the same time of day as the next ovulation, which was delayed as in the first experimental situation. Subsequent ovulations were resynchronised and followed at intervals according to the normal sequence established before the injection.

3. Injection of 0.5, 0.1 or 0.05 mg progesterone/kg between 12 and 9 h before an expected ovulation advanced the oviposition of the egg already in the uterus (shell gland) by about 3 h. The succeeding ovulation was either advanced or blocked.

4. These observations suggest that the pre‐ovulatory surge of progesterone is directly or indirectly involved in the timing of oviposition and ovulation.     

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     

Effects of Latent Infection of Stock Plants and Abundance of Thrips on the Occurrence of <Emphasis Type="Italic">Tomato spotted wilt virus </Emphasis>in Chrysanthemum Fields

DAS-ELISA proved to be reliable enough to detect a latent infection by Tomato spotted wilt virus (TSWV) in asymptomatic stock plants of chrysanthemum. A high density of Frankliniella occidentalis, the predominant vector, in the presence of latently infected stock plants resulted in a high incidence of disease in the chrysanthemum production field. The incidence of disease was low when the vector thrips were not abundant in spite of the presence of latently infected stock plants. These results suggest that an infestation of the vector thrips causes severe secondary spread of TSWV originating from latently infected stock plants in chrysanthemum production fields. Received 27 July 2001/ Accepted in revised form 27 November 2001     

Indole-3-acetic acid biosynthesis in <Emphasis Type="Italic">Aciculosporium take</Emphasis>, a causal agent of witches' broom of bamboo

 Aciculosporium take (Ascomycota; Clavicipitaceae) is a causal agent of witches' broom of bamboo plants. The symptoms of this disease are believed to be induced by plant hormones, particularly auxins. Indole-3-acetic acid (IAA) was identified in cultures of this fungus in an l-tryptophan-supplemented liquid medium. IAA production was confirmed on 30 isolates of A. take from various hosts and locations at levels up to 1 mg/l. The biosynthetic pathway of IAA in A. take culture was examined by analyzing intermediate products and by feeding experiments. The results showed that the indole-3-pyruvic acid pathway (l-tryptophan → indole-3-pyruvic acid → indole acetaldehyde → IAA) was the dominant pathway in A. take. Received: June 3, 2002 / Accepted: July 25, 2002