Abundant PrP(CWD) in tonsil from mule deer with preclinical chronic wasting disease.

A monoclonal antibody dot-blot assay was used to evaluate detergent lysates of tonsil tissue from mule deer to detect PrP(CWD), the marker for the cervid transmissible spongiform encephalopathy chronic wasting disease (CWD). Samples of formalin-fixed brain and tonsil tissues from mule deer were examined for PrP(CWD) using immunohistochemistry (IHC) with Mab F99/97.6.1, the gold standard for diagnosis of preclinical CWD. The contralateral tonsil from each of the 143 deer was prepared for confirmatory IHC and as a 10% (wt/vol) detergent lysate without purification or enrichment steps for monoclonal antibody dot-blot assay. PrP(CWD) was detected by dot-blot assay in 49 of 50 samples considered positive by IHC. Forty-eight of the positive samples were evaluated with a quantitative dot-blot assay calibrated with recombinant PrP. Tonsillar PrP(CWD) concentrations ranged from 34 to 1,188 ng per 0.5 mg starting wet weight of tissue. The abundant PrP(CWD) in mule deer tonsil will facilitate development and validation of high-throughput screening tests for CWD in large populations of free-ranging deer.     

Response of Ugandan common bean varieties to Pseudocercospora griseola and Angular leaf spot disease development in varietal mixtures

Angular leaf spot (ALS) is one of the most economically important bean diseases in Africa. One promising control option for the disease is the use of mixtures of resistant and susceptible varieties. This research evaluated (1) the reaction of farmer preferred bean varieties to ALS in the screenhouse and on-station and (2) the effect of different spatial arrangements of resistant and susceptible bean varieties on ALS disease development. For the latter, five mixture combinations and two controls were laid out in a Randomized Complete Block Design having three replicates. Analysis of variance and least significant differences (LSDs) were used to compare disease levels in both studies above. Varietal screening showed significant differences in varietal reaction to ALS. Screenhouse disease scores ranged between 0 and 5, compared to 0.7–3.9 in the field. For the varietal mixture trial, the lowest disease levels and the highest mixture efficiencies were observed for the combination of equal proportions of the susceptible and resistant varieties randomly mixed (even mixture) prior to planting. We conclude that even mixtures reduce the amount of ALS disease that develops in the season.     

Comparison of three immunodiagnostic tests for experimental Heterophyes heterophyes infection in dogs

The aim of this study was to compare the performance of three in-house diagnostic tests, i.e. counter current immunoelectrophoresis (CCIE), intradermal (ID) and indirect fluorescent immunoassay (IFI), for the diagnosis of Heterophyes infection. One hundred and twenty puppies were randomly divided into eight groups (n=15/group). Heterophyes heterophyes infections were established in these puppies by administering a dose of 50 H. heterophyes encysted metacercariae/puppy by gavage. Forty puppies of similar age and sex were divided into eight groups, of five puppies each and were used as negative controls. Sera pooled from separate infected and uninfected groups were tested against H. heterophyes antigens, weekly for 8 weeks post-infection (PI). The ID assay detected infected puppies sooner than any of the serological tests. Sero-conversion was first detected 2 weeks PI by ID assay and 1 week later by CCIE and IFI assays. ID test performed well till the end of the experiment (sensitivity and specificity: 100% and 90%, respectively). Both IFI and CCIE assays had a sensitivity of 40% and 20%, respectively for early detection of antibody, which was less sensitive than ID but both assays were more specific (100%) than the ID assay. The lowest agreement was between ID and IFI tests (40.3%), whilst the highest was observed between CCIE and IFI tests (67.2%). Of the three evaluated methods, the ID test could be recommended for scientific and laboratory diagnosis of heterophyosis in naturally infected animals. However, since none of the investigated method are optimal (i.e, offers 100% specificity and sensitivity), the choice of test employed must depend on the aim of the study.     

Low levels of haptoglobin and putative amino acid sequence in Taiwanese Lanyu miniature pigs

Porcine haptoglobin (Hp) is an acute phase protein. Its plasma level increases significantly during inflammation and infection. One of the main functions of Hp is to bind free hemoglobin (Hb) and inhibit its oxidative activity. In the present report, we studied the Hp phenotype of Taiwanese Lanyu miniature pigs (TLY minipigs; n=43) and found their Hp structure to be a homodimer (beta-alpha-alpha-beta) similar to human Hp 1-1. Interestingly, Western blot and high performance liquid chromatographic (HPLC) analysis showed that 25% of the TLY minipigs possessed low or no plasma Hp level (<0.05 mg/ml). The Hp cDNA of these TLY minipigs was then cloned, and the translated amino acid sequence was analyzed. No sequences were found to be deficient; they showed a 99.7% identity with domestic pigs (NP_999165). The mean overall Hp level of the TLY minipigs (0.21 +/- 0.25 mg/ml; n=43) determined by enzyme-linked immunosorbent assay (ELISA) was markedly lower than that of domestic pigs (0.78 +/- 0.45 mg/ml; p<0.001), while 25% of the TLY minipigs had an Hp level that was extremely low (<0.05 mg/ml). In addition, the initial recovery rate (first 40 min) in the circulation of infused fluorescein isothiocyanate (FITC)-Hb was significantly higher in the TLY minipigs with extremely low Hp levels than those with high levels. This data suggests that the low concentration of Hp-Hb complex is responsible for the higher recovery rate of Hb in the circulation. TLY minipigs have been used as an experimental model for cardiovascular diseases; whether they can be used as a model for inflammatory diseases, with Hp as a marker, remains a topic of interest. However, since the Hp level varies significantly among individual TLY minipigs, it is necessary to prescreen the Hp levels of the animals to minimize variation in the experimental baseline. The present study may provide a reference value for future use of the TLY minipig as an animal model for inflammation-associated diseases.     

Experimental observations on prophylactic and therapeutic vaccination against foot-rot in sheep

Extract

I have taken as my subject for this address the topic, “Change”, and will deal with two aspects of change, one the challenge of change due to new knowledge and techniques and second the change we must make in our attitude to other agricultural disciplines and ancillary veterinary personnel.     

Sedative effects of dexmedetomidine,dexmedetomidine–pethidine and dexmedetomidine–butorphanol in cats

The purpose of this study was to assess the clinical effects of dexmedetomidine, both alone and combined with pethidine or butorphanol, in cats. A prospective randomized blind study was performed. Thirty cats were randomly assigned to three groups of 10 animals: D: dexmedetomidine (20 μg/kg IM); DP: dexmedetomidine (10 μg/kg IM) and pethidine (2.5 mg/kg IM); DB: dexmedetomidine (10 μg/kg IM) and butorphanol (0.4 mg/kg IM). Quality of sedation, analgesia, muscle relaxation and the possibility of performing some clinical procedures were compared using a multifactorial scale. Sedation, analgesia and muscle relaxation increased progressively over time and did not differ in the three protocols. The three protocols facilitated the completion of several clinical procedures. The clinical variables studied showed a similar behaviour in the three protocols and remained close to the baseline, except for a drop in heart rate in protocol D. In conclusion, dexmedetomidine, either alone or combined with pethidine or butorphanol, offers suitable sedation, analgesia and relaxation to perform various clinical procedures in cats.