Additive and synergistic pharmacologic inhibition of equine fibrinoligase (factor XIIIa*-like) biochemical activity.

A selected group of pharmaceutical compounds were evaluated for the ability to inhibit the biochemical activity of fibrinoligase (coagulation factor XIIIa*) in pooled equine plasma. Criteria for the pharmaceuticals selected were based on the mechanism of the transglutamination biochemical reaction mediated by coagulation factor XIIa*. These criteria were complemented by recognition of the molecular configuration and chemical composition of amino acid residue side chains involved in the process of covalent fibrin monomer polymerization (cross-linking, transglutamination) mediated by this enzyme. Each pharmaceutical was evaluated individually and in combination with other potential coagulation factor XIIIa* inhibitors in an effort to detect additive and synergistic phenomenon. In this context, pharmaceuticals with a carbonylamide (eg, cefuroxime, Girard's reagent-P, prolinamide) were applied in concert with compounds with a terminal amine (eg, D-arginine, L-lysine) functional group. In concept, this method theoretically served to competitively simulate glutamine and lysine amino acid residues within strands of fibrin monomer substrate involved in phase I (carbonylamide) and phase II (terminal amine) of the transglutamination reaction (covalent fibrin monomer cross-linking). Halogen-dinitro and ethylene compounds were also evaluated because of their reported ability to inactivate enzyme systems dependent on an intact sulfhydryl group located at their biochemically active site (eg, cystine amino acid residue). This group of pharmaceutical compounds failed to inhibit the biochemical activity mediated by coagulation factor XIIIa* in equine plasma.     

Development and psychometric testing of an instrument designed to measure chronic pain in dogs with osteoarthritis

OBJECTIVE: To develop and psychometrically test an owner self-administered questionnaire designed to assess severity and impact of chronic pain in dogs with osteoarthritis. SAMPLE POPULATION: 70 owners of dogs with osteoarthritis and 50 owners of clinically normal dogs. PROCEDURES: Standard methods for the stepwise development and testing of instruments designed to assess subjective states were used. Items were generated through focus groups and an expert panel. Items were tested for readability and ambiguity, and poorly performing items were removed. The reduced set of items was subjected to factor analysis, reliability testing, and validity testing. RESULTS: Severity of pain and interference with function were 2 factors identified and named on the basis of the items contained in them. Cronbach's alpha was 0.93 and 0.89, respectively, suggesting that the items in each factor could be assessed as a group to compute factor scores (ie, severity score and interference score). The test-retest analysis revealed kappa values of 0.75 for the severity score and 0.81 for the interference score. Scores correlated moderately well (r = 0.51 and 0.50, respectively) with the overall quality-of-life (QOL) question, such that as severity and interference scores increased, QOL decreased. Clinically normal dogs had significantly lower severity and interference scores than dogs with osteoarthritis. CONCLUSIONS AND CLINICAL RELEVANCE: A psychometrically sound instrument was developed. Responsiveness testing must be conducted to determine whether the questionnaire will be useful in reliably obtaining quantifiable assessments from owners regarding the severity and impact of chronic pain and its treatment on dogs with osteoarthritis.     

Feline calicivirus

Feline calicivirus (FCV) is an important and highly prevalent pathogen of cats. It belongs to the family Caliciviridae which includes other significant pathogens of man and animals. As an RNA virus, high polymerase error rates convey upon FCV a high genome plasticity, and allow the virus to respond rapidly to environmental selection pressures. This makes the virus very adaptable and has important implications for clinical disease and its control. Being genetically diverse, FCV is associated with a range of clinical syndromes from inapparent infections to relatively mild oral and upper respiratory tract disease with or without acute lameness. More recently, highly virulent forms of the virus have emerged associated with a systemic infection that is frequently fatal. A proportion of FCV infected cats that recover from acute disease, remain persistently infected. In such cats, virus evolution is believed to help the virus to evade the host immune response. Such long-term carriers may only represent a minority of the feline population but are likely to be crucial to the epidemiology of the virus. Vaccination against FCV has been available for many years and has effectively reduced the incidence of clinical disease. However, the vaccines do not prevent infection and vaccinated cats can still become persistently infected. In addition, FCV strain variability means that not all strains are protected against equally. Much progress has been made in understanding the biology and pathogenesis of this important feline virus. Challenges for the future will necessarily focus on how to control the variability of this virus particularly in relation to emerging virulent strains and vaccination.     

Treatment of the early life stages of scallop (Pecten maximus) with antimicrobial agents; searching for an alternative to chloramphenicol

High mortality rates are often observed in rearing the early stages of the great scallop, Pecten maximus. The addition of antibacterial agents has been necessary to improve larval survival. However, as one antibacterial agent, chloramphenicol, is banned in Norway and Europe the aim of this study is to investigate alternative antibacterial agents. The therapeutic agents investigated in this study were florfenicol, oxytetracycline, oxolinic acid, neomycin and Pyceze. The mean minimum inhibitory concentration (MIC) values were determined for oxytetracycline, oxolinic acid and Pyceze against bacteria isolated from scallop larvae. Two types of treatment regime were investigated on an intermediate scale (20 L). One regime involved continuous exposure of scallop larvae to the therapeutic agent while the other involved a short exposure lasting two hours. All intermediate scale treatments were performed in parallel to large-scale production (800 L) treatment with chloramphenicol. Of the therapeutants investigated, oxolinic acid was the most effective, although only at high concentrations. The short exposure of two hours was ineffective for all therapeutics.     

Comparison of leaf and pod disease reactions of beans (Phaseolus vulgaris L.) inoculated by different methods with strains of Xanthomonas campestris pv. Phaseoli (Smith) dye

Summary Common blight disease in beans (Phaseolus vulgaris), caused by the bacterium Xanthomonas campestris pv. phaseoli, reduces crop yield and seed quality. Information is needed on the variation of leaves and pods disease reaction to strains of the bacterium after different inoculation methods. Phaseolus vulgaris cultivars Red Kidney Charlevoix, GN Harris, GN 1140, and GN Emerson were inoculated with three different strains of Xanthomonas campestris pv. phaseoli at two inoculum concentrations (10⁸ and 10⁶ bacterial cells/ml) using water soaking, multiple needle, and razor blade inoculation on leaves, and razor blade scratch, dissecting needle, and razor blade cut inoculation on pods. Differential cultivar disease reactions of leaves, pods, or both to the bacterial strains were observed in some cases. Significant interactions among cultivars, inoculation methods, strains, and inoculum concentrations (leaves) were found. A rapid leaf chlorosis developed 6 to 7 days after inoculation. Strains of bacteria did not show specificity in inducing this reaction, but rapid leaf chlorosis was associated with high inoculum concentration and with the water soaking and multiple needle methods. Another experiment was conducted to count the number of living bacterial cells deposited in the leaf tissue after inoculation by different methods. The number of bacteria deposited by water soaking or multiple needle was higher than that deposited by razor blade.Published as Paper No. 8584, Journal Series, Nebraska Agricultural Experiment Station. Research was conducted under Project No. 20–36.     

Screening techniques and sources of resistance to root diseases in cool season food legumes

Summary Soil-borne fungal diseases are among the most important factors, limiting the yield of grain legumes in many countries worldwide. Root rot, caused by Aphanomyces euteiches, Rhizoctonia solani, Fusarium solani and wilt, caused by several formae speciales of Fusarium oxysporum are the most destructive soil-borne diseases of pea, chickpea, lentil, fababean and lupin. The most effective control of these diseases is achieved through the use of resistant varieties. In this paper, recent advances in conventional and innovative screening methods for disease resistance are presented. Many grain legume accessions, which are maintained in national and international germplasm collections, have been evaluated for disease resistance and numerous resistant varieties have been released following incorporation of identified resistance genes from these sources. Recent identification of molecular markers tightly linked to resistance genes has greatly enhanced breeding programs by making marker assisted selection (MAS) possible and allowing the development of varieties with multiple disease resistance. Progress in the understanding of the biology of soil-borne fungal pathogens of grain legumes is also reviewed with particular reference to the genetic structure of their populations, diagnosis and host–pathogen interaction.     

Application of biotechnology in breeding lentil for resistance to biotic and abiotic stress

Summary Lentil is a self-pollinating diploid (2n = 14 chromosomes) annual cool season legume crop that is produced throughout the world and is highly valued as a high protein food. Several abiotic stresses are important to lentil yields world wide and include drought, heat, salt susceptibility and iron deficiency. The biotic stresses are numerous and include: susceptibility to Ascochyta blight, caused by Ascochyta lentis; Anthracnose, caused by Colletotrichum truncatum; Fusarium wilt, caused by Fusarium oxysporum; Sclerotinia white mold, caused by Sclerotinia sclerotiorum; rust, caused by Uromyces fabae; and numerous aphid transmitted viruses. Lentil is also highly susceptible to several species of Orabanche prevalent in the Mediterranean region, for which there does not appear to be much resistance in the germplasm. Plant breeders and geneticists have addressed these stresses by identifying resistant/tolerant germplasm, determining the genetics involved and the genetic map positions of the resistant genes. To this end progress has been made in mapping the lentil genome and several genetic maps are available that eventually will lead to the development of a consensus map for lentil. Marker density has been limited in the published genetic maps and there is a distinct lack of co-dominant markers that would facilitate comparisons of the available genetic maps and efficient identification of markers closely linked to genes of interest. Molecular breeding of lentil for disease resistance genes using marker assisted selection, particularly for resistance to Ascochyta blight and Anthracnose, is underway in Australia and Canada and promising results have been obtained. Comparative genomics and synteny analyses with closely related legumes promises to further advance the knowledge of the lentil genome and provide lentil breeders with additional genes and selectable markers for use in marker assisted selection. Genomic tools such as macro and micro arrays, reverse genetics and genetic transformation are emerging technologies that may eventually be available for use in lentil crop improvement.     

Breeding and genetic studies of tolerance to several bean (Phaseolus vulgaris L.) bacterial pathogens

Summary A breeding and genetic program to develop Phaseolus vulgaris bean varieties tolerant to the bacterial pathogens Pseudomonas phaseolicola, Xanthomonas phaseoli and Corynebacterium flaccumfaciens was conducted from 1962 to the present tme. The research progress is reported. Great Northern (GN) Nebraska #1, sel. 27 and PI 150414 dry bean lines were highly tolerant to races 1 and 2 of the halo blight bacterium (P. phaseolicola). The tolerant inoculated leaf, pod, and non-systemic chlorosis reactions to this bacterium were each controlled by different major genes. Coupling linkage was detected between genes controlling the leaf and systemic chlorosis reactions. GN Nebraska #1, sel. 27 and PI 207262 were tolerant to isolates (USA) of X. phaseoli, causal pathogen of common blight disease. Reaction to this bacterium was inherited quantitatively. Narrow sense heritability estimates of the disease reaction were low. Genes controlling late maturity and tolerant reaction were found to be linked in crosses with GN Nebraska #1, sel. 27 but linkage was not apparent in one cross with PI 207262. A different reaction of pod and foliage was observed in some bean lines. Susceptibility increased with the onset of plant maturity. PI 165078 was tolerant to C. flaccumfaciens and the disease reaction was simply inherited. The dry bean varieties GN Tara and GN Jules, tolerant X. phaseoli, and GN Emerson tolerant to C. flaccumfaciens and X. phaseoli were released in recent years. Breeding approaches to develop bean varieties tolerant to these bacterial pathogens is discussed.Published as Paper No. 3711, Journal Series, Nebraska Agricultural Experiment Station. Research was conducted under Project No. 20-3.