Development of PCR-based tests for the identification of North American isolates of epizootic haemorrhagic disease virus.

A serogroup-specific polymerase chain reaction (PCR) assay and isolate identification strategies (restriction endonuclease analysis (REA) and nucleotide sequencing) were developed for the detection of North American isolates of epizootic haemorrhagic disease virus (EHDV). PCR primers (EHDV-pr4, EHDV-pr5) were designed to hybridize to the L3 gene of a North American isolate of EHDV serotype 1. Total nucleic acid was extracted from preparations of infected tissue culture and PCR was performed using a cDNA-PCR kit, according to the manufacturer's specifications. The PCR assay generated a 459 base pair product from North American isolates of EHDV serotypes 1 and 2, while bluetongue virus (BTV) serotypes 10, 11, 13, and 17, and cell controls, failed to demonstrate PCR products. Slight modifications allowed for the PCR detection of EHDV-1 and -2 in white-tailed deer blood (Odocoileus virginiatus); PCR fragments were not amplified from uninfected deer blood. A number of restriction endonucleases and sequencing primers were evaluated for their utility in isolate identification experiments. Specifically, REA employing HincII and cycle sequencing with an internal primer (EHDV-1-pr3) proved most successful for identifying isolate-specific genome markers. The techniques presented are expected to prove valuable for rapid and specific detection of possible future EHDV incursions in wild and domestic animal species.     

Epidemiological investigations of trichostrongylid infections in young cattle in different parts of Norway

The occurrence of trichostrongylid nematodes in young cattle was studied in relation to climate and geographical location, by examination of herbage and faecal samples from 8 farms in 7 different regions in Norway, during 3 consecutive summers. Species of Ostertagia and Cooperia were dominant. Infective larvae, especially those of Ostertagia, were able to survive the winter in the pasture to a great extent in all the areas studied. The spring larval counts varied considerably from place to place, and from year to year, but were often at fairly high levels. This confirms previous findings that the larvae which overwinter on the pasture may be a major cause of losses. Overt clinical disease, however, was only seen at 1 farm during this investigation.The large number of larvae often seen in the autumn seems to be of less significance as a cause of disease, due to the late appearance, but is of major importance as a source of the larvae which survive the winter in the pasture and infect young cattle the following spring.There was no obvious correlation between pasture larval counts and the climate, except that there seemed to be higher counts in the warmer parts of the country, where the grazing season is also longer. The only place were both faecal egg counts and larval counts in the herbage were consistently low, was in the very far north.     

Decreased platelet function in Cavalier King Charles Spaniels with mitral valve regurgitation

With aggregometry, increased platelet activity has been reported in Cavalier King Charles Spaniels (CKCS) without mitral regurgitation (MR). In contrast, dogs with MR have been found to have decreased platelet activity. The purpose of this study was to test an easy bedside test of platelet function (the Platelet Function Analyzer [PFA-100]) to see if it could detect an increase in platelet activity in CKCS without MR and a decrease in platelet activity in CKCS with MR. This study included 101 clinically healthy dogs > 1 year of age: 15 control dogs of different breeds and 86 CKCS. None of the dogs received medication or had a history of bleeding. The PFA-100 evaluates platelet function in anticoagulated whole blood under high shear stress. Results are given as closure times (CT): the time it takes before a platelet plug occludes a hole in a membrane coated by agonists. The CT with collagen and adenosine-diphosphate as agonists was similar in control dogs (median 62 seconds; interquartile interval 55-66 seconds) and CKCS with no or minimal MR (55; 52-64 seconds). The CT was higher in CKCS with mild MR (regurgitant jet occupying 15-50% of the left atrial area) (75; 60-84 seconds; P = .0007) and in CKCS with moderate to severe MR (jet > 50%) (87: 66-102 seconds; P < .0001). CKCS with mild, moderate, and severe, clinically inapparent MR have decreased platelet function. The previous finding of increased platelet reactivity in nonthrombocytopenic CKCS without MR could not be reproduced with the PFA-100 device.     

Detection of Porcine circovirus type 2 viremia and seroconversion in naturally infected pigs in a farrow-to-finish barn

Porcine serum was assayed by 2 polymerase chain reaction (PCR) protocols (nested PCR [nPCR] and non-nested PCR) and a competitive enzyme-linked immunosorbent assay to determine when Porcine circovirus type 2 (PCV2) viremia and a rise in the serum level of PCV2-specific antibody occurred in pigs raised in a large Canadian farrow-to-finish barn. Eight serial blood samples were collected from each of 40 pigs from 5 to 156 (+/- 1.5) d of age; 6 pigs were removed from the study for various reasons at various times. Viremia was not detected in the samples collected before 72 d of age but was detected in those collected on or after 72 d: of 33 pigs, 7 (21%) had only 1 serum sample positive for PCV2 DNA by nPCR after day 72; 11 (33%) were intermittently positive by nPCR, non-nested PCR, or both between 72 and 156 d; and the remaining 15 (45%) were repeatedly positive (in 2 to 4 samples). The level of serum antibody against PCV2 declined after weaning and increased between 72 and 107 d of age, only after PCV2 was detected in serum. Our results show that PCV2 viremia persists in the presence of elevated levels of PCV2-specific antibody.     

RIP-X: Decision software to compare crosscut-first and rip-first rough mill systems

RIP-X is software which was developed to simulate the rough mill cutup of lumber. It determines the yields of the current and least-cost grade mixes for both the crosscut-first system and the rip-first system. A statistical comparison of the crosscut-first and rip-first current yields is made. The least-cost grade mix is determined by a linear programming model incorporated into the software.Validation of the RIP-X linear programming model was performed by comparing its results to results from an existing model. When the parts' lengths were restricted to the existing model's constraints, the RIP-X results did not differ significantly from the existing model's results. Comparison of current available model results with the unrestricted RIP-X model, indicated that previous models have provided sub-optimal solutions because of maximum parts' length restrictions in some lumber grades.Field tests were conducted to determine the accuracy of the crosscut-first and rip-first simulations in rough mills. The yield estimates from the crosscut-first and rip-first simulations did not differ significantly from the actual rough mill yields.     

Local vs official criteria and indicators for evaluating community forest management

Processes to develop criteria and indicators (C and I) for sustainableforest management are often expert driven. Although the long-termobjective of the Nepalese forest policy is sustainable forestmanagement, no national level C and I specifically for thishave been developed. However, the Ministry of Forests and SoilConservation awards an annual prize to successful communityforest user groups (CFUGs) based on evaluation according to10 criteria. This study explored local C and I for successfulforest management in Nepal through six meetings with CFUG members.A total of 14 local criteria and 52 indicators for awardinga prize to successful CFUGs were identified, and comparisonwith official C and I was undertaken. Most conspicuously, localC and I focus more on environmental and governance-related aspectsthan the official list, and the official list leaves much tothe discretion of the evaluator. The study concludes that developmentof a national level C and I for sustainable forest managementcould benefit from involvement of local forest users.     

Role of phospholipase A2 in the induction of drip loss in porcine muscle

The role of phospholipase A2 in the induction of drip loss from pig muscle has been investigated. In samples from porcine M. longissimus dorsi, total PLA2 activity as well as mRNA and protein levels of the group VIA iPLA2 (iPLA2-VIA) increased during the initial 4 h post-mortem period. Morphological studies of porcine muscle showed that at 4 h post-mortem, gaps had formed between muscle fibers and that the sarcolemma membrane borders appeared blurred. At the same time iPLA2-VIA protein levels were increased inside muscle fibers and at the sarcolemma. iPLA2-VIA mRNA abundance in samples from different breeds of pigs with variations in drip loss revealed no clear correlation between drip loss level and iPLA2-VIA expression. Together, these data indicate that during the post-mortem period, iPLA2-VIA expression and activity is increased at the muscle fiber membranes. PLA2 activity may affect membrane permeability and consequently the progression of drip formation in porcine muscle.