P-76 The effect of omega-3 fatty acid supplementation on cutaneous and plasma fatty acid concentrations in dogs with atopic dermatitis

The objectives of this study were to evaluate in vivo tolerance, and antimicrobial and clinical activities of a topical otic preparation containing EDTA tromethamine (Tris) and chlorhexidine digluconate 0.15% solution (Otodine^®) in dogs with chronic bacterial otitis externa. Eleven dogs were included. The affected ears were filled with the solution once daily during a 2-week period. Dogs were evaluated on days 0, 14 and 28. Three clinical parameters (exudate, erythema, pain) and three cytologic parameters ( Malassezia , cocci, rods) were scored (0–4 scale) by otoscopic and cytological examinations of otic exudate. Bacterial cultures were performed at each time point. If there were bacteria on cytological examination on day 14, the dogs were treated with the original product, with the addition of enrofloxacin (5%) applied 10 min after the original product, for a further 2 weeks. All 11 cases yielded isolates of resistant gram-negative bacteria; gram-positive bacteria were also isolated from six of 11 dogs. On day 14, six of 11 dogs were negative on culture examination; on day 28, 10 of 11 were negative and only one case had a positive culture. On day 14, clinical and microbial scores (cytology) were reduced by 54.6 and 71.1%, respectively, and by 85.7 and 94% on day 28. All cases reported good tolerance of the treatment. The results show that this ear solution was helpful in the management of chronic bacterial otitis externa in dogs and was well tolerated. There seems to be a synergistic effect of the combination of Tris-EDTA/chlorhexidine digluconate 0.15% solution, and an antimicrobial agent (enrofloxacin) against resistant gram-positive and gram-negative bacteria.
Funding: Self-funded.     

Morphology,cytochemical staining and ultrastructural characteristics of reindeer (Rangifer tarandus) leukocytes

Peripheral blood smears from four adult reindeer (Rangifer tarandus) were examined after staining with Romanowsky's stain and cytochemical stains, including alpha-napthyl butyrate esterase (alpha-NBE), Sudan black B (SBB), chloroacetate esterase (CAE) and alkaline phosphatase (ALP). Romanowsky-stained eosinophils, neutrophils, lymphocytes and monocytes resembled those of cattle, sheep and goats. Basophils had two different staining patterns with Romanowsky's stain. Basophils that we termed "grey basophils" were similar in appearance to grey eosinophils in Greyhound dogs, with medium blue-grey to lavender-grey cytoplasm containing varying numbers of clear vacuoles or granules and variable numbers of small, intensely basophilic, perinuclear granules. The second basophil staining pattern was more typical of ruminant basophils, with uniform, pale to dark basophilic cytoplasmic granules. Basophils stained positive for alpha-NBE, SBB, CAE, and ALP. Eosinophils stained positive for SBB, and were negative for alpha-NBE, CAE, and ALP. Neutrophils were negative for SBB, CAE, and ALP. Monocytes stained positive for alpha-NBE, were rarely positive for CAE and SBB, and were negative for ALP. Transmission electron microscopy revealed matrix within all granulocytes granules, including those of basophils.     

Effects of dietary cysteine on blood sulfur amino acid, glutathione, and malondialdehyde concentrations in cats

OBJECTIVE: To determine effects of dietary cysteine on blood sulfur amino acids (SAA), reduced glutathione (GSH), oxidized glutathione (GSSG), and malondialdehyde (MDA) concentrations in cats. ANIMALS: 12 healthy adult cats. PROCEDURE: Cats were fed diets with a nominal (0.50 g/100 g dry matter [DM]), moderate (1.00 g/100 g DM), or high (1.50 g/100 g DM) cysteine content in a 3 X 3 Latin square design with blocks of 8 weeks' duration. Venous blood samples were collected after each diet had been fed for 4 and 8 weeks, and a CBC and serum biochemical analyses were performed; poikilocyte, reticulocyte, and Heinz body counts were determined; and MDA, GSH, GSSG, and SAA concentrations were measured. RESULTS: Blood cysteine and MDA concentrations were not significantly affected by dietary cysteine content. Blood methionine, homocysteine, and GSSG concentrations were significantly increased when cats consumed the high cysteine content diet but not when they consumed the moderate cysteine content diet, compared with concentrations obtained when cats consumed the nominal cysteine content diet. Blood GSH concentrations were significantly increased when cats consumed the moderate or high cysteine content diet. CONCLUSIONS: Increased dietary cysteine content promotes higher blood methionine, homocysteine, GSH, and GSSG concentrations in healthy cats. CLINICAL RELEVANCE: Supplemental dietary cysteine may be indicated to promote glutathione synthesis and ameliorate adverse effects of oxidative damage induced by disease or drugs.     

Comparison of the effects of asparaginase administered subcutaneously versus intramuscularly for treatment of multicentric lymphoma in dogs receiving doxorubicin

OBJECTIVE: To determine the effectiveness and safety of asparaginase administered s.c. versus i.m. for treatment of multicentric lymphoma in dogs receiving doxorubicin. DESIGN: Prospective study. ANIMALS: 49 dogs with multicentric lymphoma. PROCEDURE: Dogs were treated with doxorubicin every 3 weeks, for a total of 5 treatments, and were given 3 weekly treatments of asparaginase, s.c. or i.m. Using high-performance liquid chromatography, mean plasma asparagine, aspartic acid, glutamine, and glutamic acid concentrations were determined in dogs before and during treatment with asparaginase (10,000 U/m2 of body surface area, once a week for 3 weeks). Asparaginase was administered s.c. in 23 dogs and i.m. in 26 dogs. Variables evaluated included time to response to chemotherapy, remission and survival times, and clinical and serum biochemical indicators of toxicoses. RESULTS: Using the World Health Organization's staging system for lymphoma, 30 dogs were in clinical stage III and 19 were in clinical stage IV. One week after asparaginase treatment, plasma asparagine concentrations were low and plasma aspartic acid, glutamine, and glutamic acid concentrations were high. Differences in plasma amino acid concentrations were not found between s.c. and i.m. groups. For dogs in clinical stage IV, i.m. administration of asparaginase significantly decreased the number of days to complete remission, compared with s.c. administration (8 vs 17 days, respectively). For dogs in clinical stage III, i.m. administration favorably increased the duration of first remission (191 vs 103 days) and survival time (289 vs 209 days). Overall, dogs treated i.m. had a faster response to chemotherapy (9 vs 15 days), a longer remission (191 vs 109 days), and a longer survival time (286 vs 198 days), compared with all dogs treated s.c. Asparaginase toxicoses were not observed regardless of the route of administration. CLINICAL IMPLICATIONS: For dogs with multicentric lymphoma that are receiving doxorubicin, i.m. treatment with asparaginase is more effective than s.c. treatment.     

Modeling approaches to estimate effective leaf area index from aerial discrete-return LIDAR

Leaf area index (LAI) has traditionally been difficult to estimate accurately at the landscape scale, especially in heterogeneous vegetation with a range in LAI, but remains an important parameter for many ecological models. Several different methods have recently been proposed to estimate LAI using aerial light detection and ranging (LIDAR), but few systematic approaches have been attempted to assess the performance of these methods using a large, independent dataset with a wide range of LAI in a heterogeneous, mixed forest. In this study, four modeling approaches to estimate LAI using aerial discrete-return LIDAR have been compared to 98 separate hemispherical photograph LAI estimates from a heterogeneous mixed forest with a wide range of LAI. Among the four approaches tested, the model based on the Beer–Lambert law with a single parameter (k: extinction coefficient) exhibited highest accuracy (r² = 0.665) compared with the other models based on allometric relationships. It is shown that the theoretical k value (=0.5) assuming a spherical leaf angle distribution and the zenith angle of vertical beams (=0°) may be adequate to estimate effective LAI of vegetation using LIDAR data. This model was then applied to six 30 m × 30 m plots at differing spatial extents to investigate the relationship between plot size and model accuracy, observing that model accuracy increased with increasing spatial extent, with a maximum r² of 0.78 at an area of 900 m². Findings of the present study can provide useful information for selection and application of LIDAR derived LAI models at landscape or other spatial scales of ecological importance.     

Damage to roots of Trifolium subterraneum L. (subterranean clover), failure of seedlings to establish and the presence of root pathogens during autumn–winter

Field‐based plant bioassays were employed to assess the potential for pre‐ and post‐emergence loss of seedlings and for root damage affecting Trifolium subterraneum L. (subterranean clover) during autumn–winter at 17 pasture sites across a broad agricultural area of temperate southern Australia. Between 9 and 93% (median 21%) of T. subterranean seedlings failed to emerge at the 14 locations where soil moisture was considered adequate for germination. Post‐emergence losses were lower (range 0–32%; median 7%). Moderate damage (lateral roots) to severe damage (taproots) was recorded on surviving test plants at all of the sites. Sublethal damage to pasture roots constitutes a potentially large, but underestimated cost to production because it was so widespread and because the damage occurs during autumn–winter when pasture yield limits stocking rate. Potential for the loss of Lolium rigidum × multiflorum (annual ryegrass) seedlings was also demonstrated at some of the sites. DNA assays for common root rot disease pathogens (Pythium irregulare, Phytopthora clandestina and Rhizoctonia) were used for the first time to construct cost‐effective profiles of fungal and oomycete pathogens at each site. These assays may be useful for indicating disease risks and guiding plant cultivar selection and appropriate use of pesticides. DNA assays for arbuscular mycorrhizal fungi were also used and have the potential to provide novel insights into the soil biology of farming systems.     

Influence of host resistance on the genetic structure of the white pine blister rust fungus in the western United States

Cronartium ribicola, the causal agent of white pine blister rust, has been devastating to five-needled white pines in North America since its introduction nearly a century ago. However, dynamic and complex interactions occur among C. ribicola, five-needled white pines, and the environment. To examine potential evolutionary influences on genetic structure and diversity of C. ribicola in western United States, population genetic analyses of C. ribicola were conducted using amplified fragment length polymorphism (AFLP) molecular markers. The fungus was sampled at six sites. Collections for two of the six sites were from separate plantings of resistant-selected western white pine and sugar pine. Heterozygosity based on polymorphic loci among populations ranged from 0.28 to 0.40, with resistant-selected plantations at the extremes. Genetic differentiation was also highest between these two populations. Principal coordinates analysis and Bayesian assignment placed most isolates that are putative carriers of virulence to major-gene resistance into a discernable cluster, while other isolates showed no clustering by site or host species. These results indicate that C. ribicola in western North America is not genetically uniform, despite its presumed single site of introduction and relatively brief residence. Moreover, major-gene resistance appears to have imposed strong selection on the rust, resulting in reduced genetic diversity. In contrast, no evidence of selection was observed in C. ribicola from hosts that exhibit only multigenic resistance.     

Summary of findings from a participant country pollen beetle questionnaire

A questionnaire was sent to all participating countries to gain some background information on rape growing and on pollen beetle ( Meligethes aeneus ). Information was sought on the use of thresholds, the availability of active substances and on the year in which resistance, if present, had been observed. A summary of the main findings of the questionnaire are presented. In total 20 countries responded and resistance in pollen beetle was reported in 12 of those countries. Information presented shows the range of active substances and doses recommended, frequency of use of insecticides on rape, and year when resistance was reported.     

Cryopreservation of Arctic charr, Salvelinus alpinus (L.), semen in various extenders and in three sizes of straw

Cryopreservation of Arctic charr Salvelinus alpinus (L.) semen was investigated using three diluents, three cryoprotectants [10% dimethyl sulphoxide (DMSO), 10% dimethylacetamide (DMA) or 20% glycerol] and three sizes of straw. The three diluents and three cryoprotectants were combined, resulting in nine extenders. One part semen was added to three parts extender, and motility was evaluated to assess the toxicity of six of the extenders. Semen in nine extenders was frozen in 0.5‐mL straws using liquid nitrogen vapour. Semen extended in 0.3 m glucose and each of the cryoprotectants was also frozen in 0.5‐mL, 1.7‐mL (flat) or 2.5‐mL straws. The freezing rate in each size of straw was measured. Fertility trials were conducted to determine the post‐thaw viability of the frozen semen. The motility of activated spermatozoa was higher in the DMA and DMSO extenders than in the glycerol extender. For the trial using 0.5‐mL straws, post‐thaw fertility results were higher for all extenders containing DMSO, or 0.3 m glucose and DMA, than for all other combinations of diluent and cryoprotectant. For the straw size comparison, the highest fertility was obtained for the 1.7‐mL straw using either DMSO or DMA and for the 2.5‐mL straw using DMSO. For all cryopreservation trials, fertility was low for extenders containing glycerol.