Diagnostics,genetic diversity and pathogenic variation of ascochyta blight of cool season food and feed legumes

Molecular diagnostic techniques have been developed to differentiate the Ascochyta pathogens that infect cool season food and feed legumes, as well as to improve the sensitivity of detecting latent infection in plant tissues. A seed sampling technique was developed to detect a 1% level of infection by Ascochyta rabiei in commercial chickpea seed. The Ascochyta pathogens were shown to be genetically diverse in countries where the pathogen and host have coexisted for a long time. However, where the pathogen was recently introduced, such as A. rabiei to Australia, the level of diversity remained relatively low, even as the pathogen spread to all chickpea-growing areas. Pathogenic variability of A. rabiei and Ascochyta pinodes pathogens in chickpea and field pea respectively, appears to be quantitative, where measures of disease severity were based on aggressiveness (quantitative level of infection) rather than on true qualitative virulence. In contrast, qualitative differences in pathogenicity in lentil and faba bean genotypes indicated the existence of pathotypes of Ascochyta lentis and Ascochyta fabae. Therefore, reports of pathotype discrimination based on quantitative differences in pathogenicity in a set of specific genotypes is questionable for several of the ascochyta-legume pathosystems such as A. rabiei and A. pinodes. This is not surprising since host resistance to these pathogens has been reported to be mainly quantitative, making it difficult for the pathogen to overcome specific resistance genes and form pathotypes. For robust pathogenicity assessment, there needs to be consistency in selection of differential host genotypes, screening conditions and disease evaluation techniques for each of the Ascochyta sp. in legume-growing countries throughout the world. Nevertheless, knowledge of pathotype diversity and aggressiveness within populations is important in the selection of resistant genotypes.     

Paecilomyces farinosus destroys powdery mildew colonies in detached leaf cultures but not on whole plants

Since 2001, several isolates of Blumeria graminis, the causal agent of cereal powdery mildew, maintained on detached leaves at the John Innes Centre, Norwich, UK, have spontaneously become infected with an unknown filamentous fungus whose mycelia have quickly overgrown the powdery mildew colonies and destroyed them completely. A total of five isolates of the contaminant were obtained and identified as Paecilomyces farinosus based on morphological characteristics and rDNA ITS sequence data. To determine whether these P. farinosus isolates can be considered as biocontrol agents (BCAs) of powdery mildews, we studied the interactions between P. farinosus and the following four powdery mildew species: B. graminis f.sp. hordei infecting barley, Oidium neolycopersici infecting tomato, Golovinomyces orontii infecting tobacco and Podosphaera fusca infecting cucumber. The powdery mildew colonies of all these four powdery mildew species were quickly destroyed by P. farinosus in leaf cultures but neither conidial suspensions nor cell-free culture filtrates of P. farinosus isolates could suppress the spread of powdery mildew infections on diseased barley, tomato, tobacco or cucumber plants in the greenhouse. It is concluded that P. farinosus cannot be considered as a promising BCA of powdery mildew infections although it can destroy powdery mildew colonies in detached leaf cultures and can be a menace during the maintenance of such cultures of cereal, apple, cucurbit and tomato powdery mildew isolates.     

ECHOCARDIOGRAPHIC PARAMETERS IN TRAINING COMPARED WITH NON-TRAINING GREYHOUNDS

Echocardiographic parameters were compared between training and non-training greyhound dogs. When indexed to body weight there was a statistically significant increase (p<0.05) in the interventricular septal thickness (systole) and when indexed to body surface area there were increased interventricular septal (systole) and left ventricular free wall measurements (systole) in training compared with non-training greyhounds. When each gender was analyzed separately and echocardiographic parameters were indexed to body size, both genders had an increase in the interventricular septal thickness (diastolic in the female, systolic in the male) in the training compared with non-training greyhounds. In male training greyhounds there was additionally an increase in the left ventricular internal dimension (systole) and free wall thickness (systole) when echocardiographic parameters were indexed to body surface area compared with non-training greyhounds (p<0.05). The results indicate that certain training greyhound echocardiographic parameters are larger than non-training greyhound echocardiographic parameters. The potential effects of training, body size and gender should be considered when interpreting echocardiographic parameters in populations of greyhounds.     

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.     

Antimicrobial use on 24 beef farms in Ontario

Understanding risk factors for antimicrobial resistance requires knowledge of antimicrobial selection pressure. The objectives of this research were to develop methodology for collecting quantitative antimicrobial use information from beef producers in Ontario, to document the types and quantities of antimicrobials reported (for a minimum of 12 mo), and to compare 2 metrics for injectable use reporting. Twenty-four volunteer beef producers were asked to complete a questionnaire, document drug use in a treatment diary, and retain empty medication containers. For injectable antimicrobials, producers recorded approximately 60% of the total use in the treatment diaries; oxytetracycline, penicillin, macrolides, florfenicol, and spectinomycin were used in the greatest quantities. Based on estimated weights of active ingredients (calculated according to number of animals exposed, duration, and average dose per day) the antimicrobials most commonly used in feed were monensin, tylosin, lasalocid, and tetracyclines. The antimicrobials most commonly used in water were lincomycin-spectinomycin, chlortetracycline, and oxytetracycline. Based on estimated weights and measured quantities, < 1% of antimicrobials used were in the Canadian category of highest importance to human medicine. A comparison of animal daily dosages to kilograms of active ingredient demonstrated that the relative ranking of use of antimicrobials varied with the chosen metric, and that further investigation into the best measure in relation to antimicrobial resistance is warranted.     

Antimicrobial use in the Alberta sheep industry.

Information regarding antimicrobial use in sheep is scarce. In 2001, a scrapie surveillance program was initiated in Alberta that also provided a mechanism for collecting other sheep health data including antimicrobial use information between April 2001 and April 2002. A major objective of this study was to describe antimicrobial use in the Alberta sheep industry. This was done by obtaining qualitative antimicrobial use information from all flocks (n = 212) providing cull ewes to the program using a brief, primarily flock-level, questionnaire. The respondents' flocks represented 13.6% of the total provincial flock in Alberta in 2001. By a substantial amount, the most frequent method of administering antimicrobials was through injection followed by in-feed, oral (liquids, pills, boluses), and in-water routes, respectively. Drug-specific use data were collected for injectable antimicrobials only, with the most commonly used antimicrobial classes being penicillins followed by tetracyclines. Producers rarely treated some or all of their flock with injectable antimicrobials after discovering an individual sick animal. Adult sheep were the most common age group treated with injectable antimicrobials and the most frequent reason for injectable antimicrobial use was mastitis followed by respiratory problems. This study provides some initial insight regarding antimicrobial use in Alberta sheep flocks. However, collection of more drug-specific data (drug type, dose/concentration, duration of treatment) for noninjectable routes of administration should be conducted in future studies. Assessing antimicrobial use in other sectors of the Alberta sheep industry (feedlots) and other provinces across Canada would also be beneficial.     

Temperature-Dependence of Lipid A Acyl Structure in Psychrobacter cryohalolentis and Arctic Isolates of Colwellia hornerae and Colwellia piezophila

Lipid A is a fundamental Gram-negative outer membrane component and the essential element of lipopolysaccharide (endotoxin), a potent immunostimulatory molecule. This work describes the metabolic adaptation of the lipid A acyl structure by Psychrobacter cryohalolentis at various temperatures in its facultative psychrophilic growth range, as characterized by MALDI-TOF MS and FAME GC-MS. It also presents the first elucidation of lipid A structure from the Colwellia genus, describing lipid A from strains of Colwellia hornerae and Colwellia piezophila, which were isolated as primary cultures from Arctic fast sea ice and identified by 16S rDNA sequencing. The Colwellia strains are obligate psychrophiles, with a growth range restricted to 15 °C or less. As such, these organisms have less need for fluidity adaptation in the acyl moiety of the outer membrane, and they do not display alterations in lipid A based on growth temperature. Both Psychrobacter and Colwellia make use of extensive single-methylene variation in the size of their lipid A molecules. Such single-carbon variations in acyl size were thought to be restricted to psychrotolerant (facultative) species, but its presence in these Colwellia species shows that odd-chain acyl units and a single-carbon variation in lipid A structure are present in obligate psychrophiles, as well.     

Evaluation of changes in microbiota after fecal microbiota transplantation in 6 diarrheic horses

The purpose of this study was to characterize the fecal microbiota of horses with acute and chronic diarrhea before and after fecal microbiota transplantation (FMT). Six client-owned horses with acute and chronic diarrhea received FMT from 2 healthy donor horses. Microbiota analysis using next-generation sequencing was performed on fecal samples collected before and 2 and 7 d after FMT. Signs of diarrhea improved in 4 horses, whereas the remaining 2 horses did not survive. There was a significant difference in the number of bacterial species between donors and recipients (P < 0.05). The Order Lactobacillales and the genera Lactobacillus, Intestinimonas, and Streptococcus were increased in the microbiota of diarrheic horses, and Saccharofermentans genus increased in healthy donors. The results suggest that FMT from the healthy donors was not effective over a 7-day period as it did not change the fecal microbiota of the diarrheic horses. Further research to improve the efficacy of FMT in horses is needed.