The sensitivity of genetic detection assays of avian influenzavirus (AIV) was compared to show that real-time polymerase chain reaction (rPCR) was 10 to 10,000 times more sensitive than conventional PCR (cPCR). Nucleotide sequences of the primers and probes for AIV genetic detection assays were compared with influenzavirus sequences registered to GenBank. A few mismatches between the nucleotide sequences of the primers and probes for the assays targeting the NP gene and those of the strains were observed. Several mismatches were observed in a case study targeting H5 genes, raising concern that sensitivity might be lower against several low pathogenic strains. Eelectrophoresis of an rPCR product was used for evaluation of the results, showing that it is rapid and sensitive enough to discriminate non-specific reaction caused in the rPCR assay.
To investigate the prevalence of Toxoplasma gondii and Neospora caninum infections, which are clinically important infectious agents of the nervous system, in dogs, we performed antibody tests for T. gondii and N. caninumn using ELISA with recombinant antigens. Of 1,979 clinically healthy household dogs living in the prefectures of Nara, Hyogo, Wakayama, and Kagawa in 2014, 7.0% were positive for T. gondii antibodies, 4.2% were positive for N. caninum antibodies, and 1.2% were positive for both. Dogs kept outdoors (11.0%) and crossbred dogs (11.3%) showed higher T. gondii antibody positive proportions than dogs kept indoors. Both positive proportions of T. gondii and N. caninum antibodies increased with age in the dogs aged one year or older. However, among the dogs under one year of age, none were T. gondii-positive, whereas 11.1% were positive for N. caninum. The above findings indicate that household dogs have a risk of T. gondii and N. caninum infections in Japan depending on how they are kept.
A quick and simple method involving glass beads for the simultaneous determination of veterinary drugs in pig muscle and kidney using liquid chromatography/mass spectrometry (LC/MS) was developed. Samples were extracted with acetonitrile, homogenized using glass beads, and cleaned with hexane. The recovery rates for 45 drugs from the pig preparation were examined. Forty-two drugs met the required validation guideline in pig kidney preparation. These results indicated that the present analysis was useful for rapid screening of veterinary drugs in livestock products.