Evidence‐Based Medicine in Canine Reproduction: Quality of Current Available Literature

The objective of this study was to evaluate the quality of published literature on reproduction in dogs. A systematic search in online databases revealed 287 papers that met the inclusion criteria. For evaluation a questionnaire comprising 40 criteria regarding materials and methodology, study design, statistics, presentation and information content, applicability and conclusions was developed. In a pre‐test including seven independent scientists the applicability and explanatory power of the questionnaire and its results were validated. Out of 287 publications evaluated, 90 (31.4%) were classified as clinical trials. The remaining 197 publications were case reports or contained information based on personal experience. Not a single meta‐analysis was found. Sixty (66.7%) of the 90 clinical trials included a control group. Randomization was conducted in 23 and blinding in eight articles respectively. In total five articles were determined as randomized, controlled and blinded clinical trials. Information content of the publications was variable concerning details on included animals, type or dosage of used remedies or conducted interventions. For example, in 99.7% of the articles, the exact number of animals was given, but in 79.8%, housing and feeding of the animals were not described. Statistical procedures of clinical trials were determined adequate in 55.6%. However, the data of 67.9% of the articles were evaluated to be not sufficient to draw valid conclusions. This study revealed evidence of deficits in the field of canine reproduction. The demand for more high quality clinical research is obvious. Requisite for the further implementation of the evidence‐based veterinary medicine is an improvement of the quantity and the quality of well‐designed, conducted and reported clinical trials. The practitioner should always assess the quality of information before implementing results into practice to provide best available care for the animals.     

Genetic characterization by fluorescent AFLP of Pseudomonas savastanoi pv. savastanoi strains isolated from different host species

The genetic diversity of 71 Pseudomonas savastanoi pv. savastanoi strains isolated from different host species and from diverse geographical regions was determined by fluorescent amplified fragment length polymorphism (f-AFLP) analysis. The study was carried out using three different selective primer combinations. Strains of P. syringae pv. syringae , P. syringae pv. phaseolicola , P. syringae pv. glycinea , P. syringae pv. tagetis and P. amygdali were also included as outgroups. Based on cluster analysis of f-AFLP data, all P. savastanoi pv. savastanoi strains showed a high degree of similarity, grouping in a cluster and forming a taxon clearly separate from outgroup strains. AFLP analyses failed to support placing strains of P. savastanoi pv. savastanoi , P. syringae pv. phaseolicola and P. syringae pv. glycinea in the same species. Strains of P. savastanoi pv. savastanoi formed subclusters that correlated with the host species. Strains identified within these subclusters were related to the geographical region where the strains were isolated. Strains of P. savastanoi pv. savastanoi from olive were divided into two subclusters. Strains from oleander were differentiated from those from ash and were divided into two additional subclusters, distinct from olive strains. Three strains isolated from jasmine showed a high level of similarity among them but, at a lower Dice similarity coefficient, were linked to a subcluster including olive strains. Finally, two strains isolated from privet were similar to strains from olive and were included in the same subcluster.     

Reduced efficacy and enhanced degradation of carbetamide after repeated application in Australia

Following reports of poor weed control, the effectiveness of repeated applications of carbetamide atid propyzamide in controlling Lolium rigidum Gaud, (annual ryegrass) was examined under field conditions. Repeated applications of carbetamide or propyzamide were less effective in controlling L. rigidum than an initial application. A single application of carbetamide or propyzamide was sufficient approximately to halve the efficacy of a subsequent treatment with the satne herbicide. Laboratory incubations of soil linked the poor performance of carbetamide upon repeated application to an enhanced deg-radation rate. We conclude that the repeated application of carbetamide or propyzamide results in less effective weed control under field conditions and that this can be partially avoided by rotation between carhetamide and propyzamide.     

Inhibition of telomerase in canine cancer cells following telomestatin treatment

Telomere shortening in normal somatic cells has been proposed as a major barrier to unlimited cellular proliferation. Telomerase is an enzyme capable of maintaining telomere length, and thus bypassing this barrier. In human beings, telomerase activity is restricted to cancer cells and cells of stem or germ cell lineages. Dogs represent a potentially useful clinical model for the development of telomerase‐based therapies because telomerase activity is also restricted to cancer cells and stem cells in this species. We examined the ability of telomestatin to inhibit telomerase activity in telomerase‐positive D17 and CMT7 canine cancer cell lines. At a concentration of 2 μM, telomestatin treatment resulted in a decrease in telomerase activity, telomere shortening, growth inhibition and apoptosis in telomerase‐positive cancer cells. These effects were not seen in telomerase‐negative skin fibroblasts or negative controls. These results confirm that telomestatin specifically inhibits telomerase activity in canine cancer cells and strengthens the usefulness of dogs as a model for testing telomerase‐based therapies.     

A molecular marker linked to the Prv1 gene that confers resistance to Papaya ringspot virus-type W in melon

Papaya ringspot virus‐type W (PRSV‐W) is the most prevalent and important viral pathogen of cucurbits in Brazil. It can be effectively controlled by the incorporation of genetic resistance into susceptible melon cultivars. The present study identified amplified fragment length polymorphic (AFLP) markers linked to the PRSV‐W resistance Prv¹ allele. The susceptible yellow‐fleshed melon‐breeding line AF426^prv1 and its nearly isogenic‐resistant line AF426^Prv1, which carries the Prv¹ allele resident in the Indian cantaloupe U.S. Plant Introduction (PI) 180280, were screened for AFLP marker polymorphisms. Of 30 251 AFLP loci, only three were polymorphic between the nearly isogenic lines. Segregation analyses for these three polymorphic markers and the Prv¹ allele using a BC₁ population of 197 plants indicated close linkage (0.5% recombination frequency) between marker EK190 (HindIII‐CGA and MseI‐GTG; 190 bp) and Prv¹. Thus, EK190 might be a useful marker in breeding programmes aiming to develop melon cultivars resistant to PRSV‐W. The other two markers are closely linked to each other, but distantly linked to Prv¹.     

Microbial biomass and respiratory activity in soil aggregates of different sizes from three beechwood sites on a basalt hill

We studied the effects of aggregates of different sizes on the soil microbial biomass. The distribution of aggregate size classes (<2, 2–4, 4–10, >10 mm) in the upper mineral soil horizon (A_h layer) was very different in three sites (upper, intermediate, lower) in a beechwood (Fagus sylvatica) on a basalt hill (Germany). Aggregates of different sizes (<2, 2–4, 4–10 mm) contained different amounts of C and N but the C:N ratios were similar. C and N contents were generally higher in smaller aggregates. The maximum initial respiratory response by microorganisms in intact aggregates and in aggregates passed through a 1-mm sieve declined with the aggregate size, but the difference was more pronounced in intact aggregates. Disruption of aggregates generally increased this response, particularly in 4- to 10-mm aggregates in the lower site. Basal respiration differed strongly among sites, but was similar in each of the aggregate size classes. Aggregate size did not significantly affect the specific respiration (g O₂ g^–1 microbial C h^–1) nor the microbial: organic C ratio, but these parameters differed among sites. Microbial growth was increased strongly by passing the soil through a 1-mm sieve in each of the aggregate materials. The growth of microorganisms in disrupted aggregates was similar, and the effect of aggregate disruption depended on the growth of microorganisms in intact aggregates.     

Nucleus substitution of Brassica japonica Sieb. with Raphanus sativus L. and its resultant chlorophyll deficiency

Summary Nucleus substitution of Brassica japonica (2n=20) with Raphanus sativus (2n=18) was carried out by means of repeated backcrossing of Brassicoraphavus (2n=37) to R. sativus as a pollen donor. In the course of nucleus substitution, chlorophyll deficiency appeared. Plants with more than 28 chromosomes, like their parents, had green leaves and those with 24 to 26 chromosomes had yellowish green ones. Almost all plants with 18 to 23 chromosomes showed yellow or whitish yellow. The R. sativus with B. japonica cytoplasm (2n=18) was obtained after four successive backerosses. The completely substituted R. sativus showed the same fertility as the true R. sativus used as a recurrent parent. It is assumed that the chlorophyll deficiency is caused by disharmony between the B. japonica cytoplasm and the R. sativus nucleus. The chlorophyll deficiency is discussed in comparison with male sterility or other characters which sometimes occur in alloplasmic Raphanus and Brassica species.