Rabies virus infection in Aedes pseudoscutellaris cells: a study on receptorial structures

Rabies virus is able to infect in vitro a wide range of homeothermic and poikilothermic cells but little is known about its multiplication in arthropod cells. In this research the infection of rabies virus in Aedes pseudoscutellaris cells, a mosquito cell line susceptible to mosquito-borne viruses, was studied. After 60 days of incubation at 26 degrees C up to 70% of infected cells showed the synthesis of both viral nucleocapsid and envelope antigens, although viral yield and cell damage could not be detected. Research performed in order to investigate the role of membrane carbohydrate moieties in rabies virus-mosquito cell interaction suggested the participation of galactose and N-acetylglucosamine whereas sialic acid, known to be a rabies binding site in many homeothermic cell lines, was not involved.     

Health of non-ambulatory, non-injured pigs at processing

Loss of pigs during or after transport is a welfare concern, but also an economic concern for producers. Transport losses include animals that are dead on arrival (DOA) at the plant, pigs that are injured, and pigs which are not obviously injured but unwilling or unable to walk (non-ambulatory, non-injured or NANI). The objective of this research was to assess the health of non-ambulatory, non-injured (NANI) pigs relative to control pigs at the processing plant by looking at a range of measures, including complete blood chemistry, anatomy, and pathology to determine potential factors associated with pigs going down. Data were collected from NANI and control pigs at five plants in the midwest USA. Feet and legs and internal organs were inspected and the severity of the pathology scored. Alveolar macrophages were collected and counted. Blood was collected for analysis of hematology, blood chemistry and cortisol concentrations. Titers to common porcine respiratory viruses were measured in pigs from one plant. Hoof and pad problems did not differ overall between control and NANI pigs, however the percentage of severe foot problems was greater (P < 0.05) in NANI compared with control pigs at plants A and E. The percentage of total ulcers, rhinitis, and empty stomachs differed (P < 0.05) between control and NANI pigs at individual plants, but not overall. Blood hematology and chemistry differed (P < 0.05) between NANI and control pigs. Cortisol concentrations did not differ between NANI and control pigs. Titers to swine influenza virus (SIV) H1N1 and H3N2 and porcine circovirus (PCV) were lower (P < 0.01) among NANI compared with control pigs. However, more (P < 0.01) NANI pigs were positive for SIV H1N1 and H3N2 compared with control pigs. Blood hematology, chemistry, and pathology indicate a large difference between NANI and controls pigs. No single health problem was higher among NANI pigs compared to plant-matched control pigs. Rather, several problems appear to contribute to pigs becoming NANI which may differ from one plant too another.     

Assessment of brachial plexus blockade in chickens by an axillary approach

ObjectiveTo assess the brachial plexus block in chickens by an axillary approach and using a peripheral nerve stimulator.Study designProspective, randomized, double-blinded study.AnimalsSix, 84-week old, female chickens.MethodsMidazolam (1 mg kg⁻¹) and butorphanol (1 mg kg⁻¹) were administered into the pectoralis muscle. Fifteen minutes later, the birds were positioned in lateral recumbency and following palpation of the anatomic landmarks, a catheter was inserted using an axillary approach to the brachial plexus. Lidocaine or bupivacaine (1 mL kg⁻¹) was injected after plexus localization by the nerve stimulator. Sensory function was tested before and after blockade (carpus, radius/ulna, humerus and pectoralis muscle) in the blocked and unblocked wings. The latency to onset of motor and sensory block and the duration of sensory block were recorded. A Friedman nonparametric one-way repeated-measures anova was used to compare scores from baseline values over time and to compare the differences between wings at each time point.ResultsA total of 18 blocks were performed with a success rate of 66.6% (12/18). The latency for motor block was 2.8 ± 1.1 and 3.2 ± 0.4 minutes for lidocaine and bupivacaine, respectively. The latencies for and durations of the sensory block were 6.0 ± 2.5 and 64.0 ± 18.0 and 7.8 ± 5.8 and 91.6 ± 61.7 minutes for lidocaine and bupivacaine, respectively. There was no statistical difference between these times for lidocaine or bupivacaine. Sensory function was not abolished in nonblocked wings.Conclusions and clinical relevanceThe brachial plexus block was an easy technique to perform but had a high failure rate. It might be useful for providing anesthesia or postoperative analgesia of the wing in chickens and exotic avian species that have similar wing anatomy.     

Descriptive spatial epidemiology of subclinical Salmonella infection in finisher pig herds: application of a novel method of spatially adaptive smoothing

We describe the spatial epidemiological features of the 6.8 million meat-juice serological tests that were conducted between 1995 and 2004 as part of the Danish swine Salmonella control programme. We investigated pig and farm density using edge-corrected kernel estimations. Pigs were aggregated at the county level to assess county-level risk, and then we investigated farm-level risk by giving farms a case or non-case label using a cut-off of 40% of pigs positive. Conditional probability surfaces, correcting for the underlying population at risk, were produced for each year of the study period using a novel kernel estimator with a spatially adaptive smoothing bandwidth. This approach improves on previous methods by allowing focussed estimation of risk in areas of high population density while maintaining stable estimates in regions where the data are sparse. Two spatial trends in the conditional probability of a farm being a case were evident: (1) over the whole country, with the highest risk in the west compared to the east; and (2) on the Jutland peninsula with the highest risk in the north and south. At the farm-level a consistent area of risk was the south-west of Jutland. Case farms tended to aggregate indicating spatial dependency in the data. We found no association between pig or farm density and Salmonella risk. We generated hypotheses for this spatial pattern of risk and we conclude that this spatial pattern should be considered in the development of surveillance strategies and as a basis for further, more detailed analyses of the data.     

Prevalence and distribution of radiographically evident lesions on repository films in the hock and stifle joints of yearling Thoroughbred horses in New Zealand

AIM: To characterise the prevalence and distribution of radiographic changes in the hocks and stifles of Thoroughbred yearling colts and fillies in New Zealand and compare them with other populations of young horses. METHODS: Repository radiographs taken in New Zealand for the 2003-2006 Thoroughbred national yearling sales were evaluated by two individual readers. The distribution of radiographic changes was classified as left side only, right side only, or bilateral. Lesions were categorised by type, location, and sex of the yearling. Complete sets of hock and stifle radiographs of 1,505 yearlings were evaluated. RESULTS: Osteophytes or enthesophytes were seen radiographically in the distal tarsal joints of 460/1,505 (31%) horses. Osteochondrosis was seen in the tibiotarsal joint of 66/1,505 (4%) horses, and in the femoropatellar joint of 40/1,505 (3%) horses. Radiographic lucency in the distal or axial aspect of the medial femoral condyle was seen in 247/1,505 (16%) horses, and lucencies consistent with subchondral cyst-like lesions were seen in 26/1,505 (2%) horses. No significant difference was seen in the proportion of colts and fillies with radiographic changes in the hock or stifle. The prevalence of osteochondrosis and subchondral cyst-like lesions in the stifles of the yearlings examined were similar to those reported in Thoroughbred yearling sale horses in the United States of America (USA). The prevalence of changes in the distal tarsal joints was similar to those reported in Standardbred and Thoroughbred yearlings from Scandinavia and the USA. CONCLUSIONS: There was no significant difference in the prevalence of osteochondrosis in the hock and stifle, lucencies in the distal medial femoral condyle, or radiographic changes in the distal tarsal joints between colts and fillies. CLINICAL RELEVANCE: Establishment of the normal prevalence and distribution of radiographic changes in the hocks and stifles of Thoroughbred yearlings in New Zealand will allow comparison with populations of young horses in other countries. Knowledge of the normal prevalence will assist veterinarians to identify abnormally high or low prevalences on individual farms, to further investigate the pathogenesis of the lesions.     

Magnetic Resonance Imaging Findings in Dogs with Confirmed Shoulder Pathology

Objective— To evaluate the diagnostic potential of magnetic resonance imaging (MRI) compared with a reference standard, arthroscopic and/or open surgery, in dogs with soft tissue shoulder pathology. Study Design— Retrospective study. Animals— Dogs (n=21). Methods— Magnetic resonance (MR) images were retrospectively evaluated in 21 dogs that had surgically identified soft tissue shoulder pathology. The musculotendinous units of the biceps, infraspinatus, teres minor, supraspinatus, subscapularis, and the medial and lateral glenohumeral ligaments (MGHL and LGHL) were graded as either normal or abnormal. Abnormal structures were further classified as being either inflamed, partially torn, or fully torn. Impingement of the biceps tendon was also evaluated. Results were reported in terms of agreement and concordance between MRI findings and surgical findings. Agreement was defined as the percentage of times MRI findings concurred with surgical findings with respect to a structure being either normal or abnormal. Concordance was defined as the percentage of times MRI concurred with the exact surgically assessed pathology when abnormality was identified. Results— The findings were biceps tendon: 90% agreement with 100% concordance; subscapularis: 95% agreement with 62% concordance; MGHL: 84% agreement with 83% concordance; LGHL: 88% agreement with 100% concordance; infraspinatus: 100% both agreement and concordance; biceps tendon impingement: 90% agreement with 100% concordance. Conclusions— Soft tissue abnormalities of the canine shoulder were readily identified on preoperative MR images. Clinical Relevance— MRI shows great potential as a diagnostic tool in the evaluation of canine shoulder disease.     

Optimization and validation of a flow cytometric method for immunophenotyping peripheral blood lymphocytes from cynomolgus monkeys (Macaca fascicularis)

BACKGROUND: Guidelines published by the Food and Drug Administration and Center for Human Medicinal Products describe the need to assess immunotoxic effects in nonclinical studies that evaluate drug toxicity, including the use of immunophenotyping to measure immunotoxicity. We are not aware of previous studies, however, that have validated methods for immunophenotyping peripheral blood lymphocyte subsets in whole blood samples from cynomolgus monkeys. OBJECTIVE: The purpose of this study was to optimize and validate a flow cytometric assay for immunophenotyping lymphocytes in the peripheral blood of cynomolgus monkeys. METHODS: A series of prevalidation experiments were done to determine optimal reagents, volumes, timing, and other procedural details of the flow cytometric assay. Using the optimized method, we then determined precision, interindividual variation, laboratory-to-laboratory variability, and sample stability. Stabilized human blood was used as a positive control for staining, processing, and analysis. The percentage and number of pan-T cells (CD3+), T-helper cells (CD3+4+), T cytotoxic/suppressor cells (CD3+8+), natural killer cells (CD3-16+), and B-cells (CD3-20+) were determined in 146 male and 140 female, clinically healthy monkeys and reference intervals were calculated. RESULTS: By doing 4-color staining with a lyse-wash method, intra- and interassay precision were <5% for all lymphocyte subsets. Variability between technicians and laboratories was minimal (CVs<3%). Samples were stable for up to 24 hours after staining and fixing. CONCLUSIONS: The validated method is extremely robust and can be performed under good laboratory practice conditions to support nonclinical studies. Reference intervals for lymphocyte subsets were similar to those previously reported.     

Assessment of in vitro interferon-gamma responses from peripheral blood mononuclear cells of cattle infected with Mycoplasma mycoides ssp. mycoides small colony type

Contagious bovine pleuropneumonia (CBPP) is a lung disease caused by the bacterial pathogen Mycoplasma mycoides ssp. mycoides small colony type (MmmSC). It has been spreading due to a number of factors including poor vaccine efficacy and poor sensitivity of current diagnostic tests. The purpose of this study was to assess interferon gamma (IFN-gamma) release after stimulation of peripheral blood mononuclear cells (PBMC) from experimentally infected cattle. PBMC collected from 15 artificially infected animals were incubated with different concentrations of total MmmSC antigen. After 72h of incubation the IFN-gamma release was measured and found to be elevated in 11 animals. We did not observe a correlation between IFN-gamma release of animals with and without pathomorphological gross lesions. Therefore, our data do not confirm a role for CD4 T-lymphocytes in protection, since there is no correlation between IFN-g secretion (supposed to be mainly derived from CD4 T-cells) and disease severity. Additionally, we applied immunocytochemistry on affected lung tissue and detected no build up of T-lymphocytes (CD4 T-cells, CD8 T-cells) but a high presence of myeloid cells.     

Serological evidence of USUTU virus occurrence in north-eastern Italy

In the recent years, USUTU virus (USUV), a flavivirus of the Japanese encephalitis virus complex, has been reported in Central Europe. As part of a systematic surveillance programme to monitor possible entrance and/or circulation of vector-borne viruses, since 2001, sentinel-chicken flocks were placed throughout the Italian territory nearby areas considered at risk of virus introduction. They have been periodically checked for the presence of antibodies against flaviviruses by indirect ELISA, plaque reduction neutralization test for USUTU, West Nile and tick-borne encephalitis viruses. In July 2007, a sentinel chicken in a flock of 20 animals located within the Ravenna province seroconverted to USUV reaching neutralizing titres up to 1:5120. A second chicken seroconverted to the same virus 2 months later. Although no virus was rescued from these animals and from wild or farm birds sampled in the area, these results still provided evidence of the circulation of USUV in north-eastern Italy.