Oligonucleotide fingerprint analysis of the genomes of two variants of bluetongue virus serotype 2 isolated in the U.S.A.

The genome segments of two electrophoretically distinct variants of bluetongue virus (BTV) Serotype 2 (Ona A and Ona B) from the U.S.A. were analyzed by double-dimension gel electrophoresis of RNase T1 produced oligonucleotides. Segments 1, 4, 5, 6, 7 and 10 were examined individually after separation by SDS-PAGE; and Segments 2 and 3, and 8 and 9, which were difficult to resolve, were fingerprinted as pairs. The Ona A and Ona B strains appeared to be closely related since corresponding segments were comparable, sharing 53–89% of the large oligonucleotides counted. Since the strains with the Ona A electropherotype preceded Ona B infection in Florida, U.S.A. and since Ona A was indistinguishable from the early African isolate of Serotype 2, Ona B was thought to be a variant of an Ona A strain. These data tend to support the hypothesis that Ona B could have evolved from Ona A as the result of point mutations or genetic drift.     

Using an independent quality control software program, EZ Runs, to monitor quality control procedures for a bench-top coagulation analyzer

BACKGROUND: Comprehensive quality control (QC) procedures are necessary to ensure accurate analytic method performance. Highly automated systems typically have inherent QC programs that facilitate performance and maintenance of QC procedures; however, for bench-top analyzers that lack internal systems, independent QC programs must be used. OBJECTIVE: The goal of this study was to evaluate the adaptability of an independent QC program, EZ Runs (Westgard QC Inc, Madison, WI, USA), to the maintenance of QC procedures for a mechanical, bench-top coagulation unit and to compare the results with our current, manual, QC method in a qualitative way. METHODS: A QC application file for activated partial thromboplastin time (aPTT) performed on a STart4 (Diagnostica Stago, Parsippany, NJ) was created in EZ Runs. Results were recorded and interpreted using this software package as well as the current, manual, QC method. RESULTS: EZ Runs was adaptable to QC monitoring for the bench-top analyzer, and the program permitted identification of both random and systematic errors not detected by the manual QC system. CONCLUSIONS: EZ Runs improved the performance and maintenance of QC procedures for this bench-top coagulation analyzer. The results indicated the need to improve staff training in assay performance and QC interpretation. In addition, use of the software program indicated that a multirule QC design was needed to monitor assay performance.     

Transmission of Phytophthora ramorum in Mixed-Evergreen Forest in California

ABSTRACT During 2001 to 2003, the transmission biology of Phytophthora ramorum, the causal agent of sudden oak death, was studied in mixedevergreen forest, a common forest type in northern, coastal California. Investigation of the sources of spore production focused on coast live oak (Quercus agrifolia) and bay laurel (Umbellularia californica), dominant hosts that comprised 39.7 and 46.2% of the individuals at the study site, respectively. All tests for inoculum production from the surface of infected coast live oak bark or exudates from cankers were negative. In contrast, sporangia and chlamydospores were produced on the surface of infected bay laurel leaves. Mean number of zoospores produced from infected bay laurel leaves under natural field conditions during rainstorms was 1,173.0 +/- SE 301.48, and ranged as high as 5,200 spores/leaf. P. ramorum was recovered from rainwater, soil, litter, and streamwater during the mid- to late rainy season in all 3 years of the study. P. ramorum was not recovered from sporadic summer rains or soil and litter during the hot, dry summer months. Concentrations of inoculum in rainwater varied significantly from year to year and increased as the rainy season progressed for the two complete seasons that were studied. Potential dispersal distances were investigated for rainwater, soil, and streamwater. In rainwater, inoculum moved 5 and 10 m from the inoculum source. For soil, transmission of inoculum was demonstrated from infested soil to bay laurel green leaf litter, and from bay laurel green leaf litter to aerial leaves of bay laurel seedlings. One-third to one-half of the hikers tested at the study site during the rainy season also were carrying infested soil on their shoes. In streamwater, P. ramorum was recovered from an unforested site in pasture 1 km downstream of forest with inoculum sources. In total, these studies provide details on the production and spread of P. ramorum inoculum in mixed-evergreen forest to aid forecasting and managing disease transmission of this environmentally destructive pathogen.     

Sensitivities of three county health department surveillance systems for child-related dog bites: 261 cases (2000)

OBJECTIVES: To determine incidence of child-related dog bites and sensitivities of 3 county health department dog-bite surveillance systems. DESIGN: Retrospective study. STUDY POPULATION: Child-related dog-bite data obtained from surveillance systems of 3 counties in Georgia in the year 2000. PROCEDURE: To characterize the sensitivity of health department dog-bite surveillance systems, 9 other potential sources of dog-bite records that matched records by victim name, age, gender, and incident date were evaluated. The number of reported bites and the most productive sources for identifying additional cases were determined. The Chandra Sekar-Deming capture-recapture method was used to estimate the number of unreported bites, and estimates of dog-bite incidence rates were made. RESULTS: 40, 36, and 185 dog bites were reported in the 3 counties, respectively. Capture-recapture calculations estimated an additional 9, 5, and 128 dog bites in these counties, respectively. Local health departments recorded 45.5% to 82.5% of dog bites. Local hospital emergency departments, police departments, and a rabies-testing laboratory received additional reports. Among these data sources, local hospital emergency department records were the best source for identifying additional cases. CONCLUSIONS AND CLINICAL RELEVANCE: Dog bites are a preventable cause of childhood injuries, and surveillance is a critical tool for tracking childhood dog bites in the community. Counties should use combined data from local health departments, local hospital emergency departments, and police departments to implement or revise dog-bite prevention programs.     

Application of multiple shrinkage methods to genomic predictions

New challenges have arisen with the development of large marker panels for livestock species. Models easily become overparameterized when all available markers are included. Solutions have led to the development of shrinkage or regularization techniques. The objective of this study was the application and comparison of Bayesian LASSO (B-L), thick-tailed (Student-t), and semiparametric multiple shrinkage methods. The B-L and Student-t methods were also each analyzed within a single shrinkage and a multiple shrinkage framework. Simulated and real data were used to evaluate each method's performance. Real data consisted of SNP genotypes of 4,069 Holstein sires. Traits included in analysis of real data were milk, fat, protein yield, and somatic cell score. The performance of each model was compared based on correlations between true and predicted genomic predicted transmitting abilities. Model performance was also compared with the performance of routinely used methods such as Bayes-A and GBLUP through cross-validation techniques. When using simulated data regardless of shrinkage framework, shrinkage models outperformed genomic BLUP (GBLUP). The average advantage of shrinkage models ranged from 1% to approximately 8% depending on the prior specification. When analyzing real data, shrinkage models slightly outperformed GBLUP for most traits. Shrinkage models were better able to model traits for which 1 or more SNP of large effect have been identified. Overall, results suggested a relatively small advantage in multiple shrinkage models. Multiple shrinkage methods could represent a useful alternative to current methods of prediction; however, their performance in a variety of scenarios needs to be investigated further.     

Comparative evaluation of live marker vaccine candidates "CP7_E2alf" and "flc11" along with C-strain "Riems" after oral vaccination

Due to the tremendous socio-economic impact of classical swine fever (CSF) outbreaks, emergency vaccination scenarios are continuously under discussion. Unfortunately, all currently available vaccines show restrictions either in terms of marker capacities or immunogenicity. Recent research efforts were therefore directed at the design of new modified live marker vaccines. Among the most promising candidates the chimeric pestiviruses "CP7_E2alf" and "flc11" were identified. Within an international research project, these candidates were comparatively tested in challenge experiments after a single oral vaccination. Challenge infection was carried out with highly virulent CSF virus strain "Koslov", 14 or 21 days post vaccination (dpv), respectively. Safety, efficacy, and marker potential were addressed. All assessments were done in comparison with the conventional "gold standard" C-strain "Riems" vaccine. In addition to the challenge trials, multiple vaccinations with both candidates were performed to further assess their marker vaccine potential. All vaccines were safe and yielded full protection upon challenge 21 days post vaccination. Neither serological nor virological investigations showed major differences among the three vaccines. Whereas CP7_E2alf also provided clinical protection upon challenge at 14 days post vaccination, only 50% of animals vaccinated with flc11, and 83% vaccinated with C-strain "Riems" survived challenge at this time point. No marked differences were seen in protected animals. Despite the fact that all multiple-vaccinated animals stayed sero-negative in the accompanying marker test, the discriminatory assay remains a weak point due to delayed or inexistent detection of some of the vaccinated and subsequently infected animals. Nevertheless, the potential as live marker vaccines could be confirmed for both vaccine candidates. Future efforts will therefore be directed at the licensing of "Cp7_E2alf" as the first live marker vaccine for CSF.     

Physiological Predictors in Calves of Dairy Breeds: Part 1. Genetic Parameters of Basal and Induced Growth Hormone Secretion

Abstract

Genetic variation in plasma growth hormone (GH) concentration before and after GRF (growth hormone releasing factor) stimulation was studied in young bulls (N=284) and heifers (N=212), the progeny of 53 sires of four dairy and dual-purpose breeds (Danish Jersey, Red Dane, Danish Friesian and Danish Red and White). Male and female calves were reared, fed and tested on separate experimental stations; thus sex, station and feeding were completely confounded effects. The animals were tested at about 9 months of age, after a 24 h fast. GH concentration was measured in serial plasma samples collected for 1 h before and 1 h following intravenous administration of 2.0 ug synthetic GRF(1–29)NH₂/kg live weight^>0.75. Prior to statistical analysis, concentrations were log_e-transformed. Response variables were BASELINE (mean GH in -15, -5 and 0 min samples) and PEAK (mean GH in 10, 15 and 20 min samples). A statistical model taking at least three generations of ancestral relationships into account was used to estimate variance and covariance components for traits in male and female calves by use of restricted maximum likelihood methods.

Heritability of BASELINE was low (0.04 ± 0.12) in males but high in females (0.60 ± 0.16). The heritability of PEAK was high in both sexes (males, 0.42 ± 0.16; females, 0.60 ± 0.16). Genetic correlations between the same trait measured in males and females were low for BASELINE (r _g = 0.32±0.55) but high for PEAK (r _g = 0.82±0.15). Within sex, BASELINE and PEAK were both highly genetically correlated (males, r _g=0.62 ± 0.40; females, r _g= 1.00 ± 0.07).

We conclude that growth hormone concentration is a highly heritable trait in juvenile cattle of both sexes, and that GRF stimulation is benificial to the uncovering of genetic differences among animals.     

Lack of detection of circulating skin-specific IgE autoantibodies in dogs with moderate or severe atopic dermatitis

Human patients with atopic dermatitis (AD) commonly exhibit IgE reactivity to cutaneous self-antigens. The presence of serum IgE autoantibodies appears to correlate with disease severity, and it is suspected to reflect or contribute to tissue damage. The objective of this study was to determine whether IgE autoantibodies specific for cutaneous antigens could be detected in the serum of dogs with AD. Serum was collected from 19 dogs with untreated moderate to severe AD and four specific-pathogen free (SPF) dogs. Indirect immunofluorescence was performed using normal canine skin collected at four different locations (concave ear, nose, medial thigh and lateral thorax), while Western immunoblotting was done using normal canine ear pinna epidermal and dermal extracts and reducing conditions. In both methods, IgE was detected using a monoclonal antibody specific for heat stable epitopes of canine IgE. At 1:10 dilution, specific IgE autoantibodies against cutaneous autoantigens were not detected, with either method, in AD and SPF canine sera. Either IgE autoreactivity is not associated with moderate to severe AD in dogs, or the methods employed herein were not sensitive enough to permit IgE autoantibody detection.