Michiko Naito

An indirect enzyme-linked immunosorbent assay (ELISA) was developed for a screening test to detect antibodies against classical swine fever virus (CSFV). Viral glycoproteins, which were purified from swine kidney cells infected with CSFV ALD/A76 strain by the immunoaffinity purification using monoclonal antibody against E2 protein, were adsorbed on a microtiter plate as the antigen for the antibody detection. Each antibody titer of serum sample was expressed as a sample per positive value calculated with optical absorbance of each sample and that of a positive control. The advantage of this ELISA is its higher sensitivity: most sera containing more than 4 neutralization titers were determined to be positive. This ELISA is unable to discriminate between antibodies against CSFV and those against other ruminant pestiviruses, therefore positive sera in this ELISA should be evaluated by a cross-neutralization test using CSFV, bovine viral diarrhea virus, and border disease virus. Taken together, the indirect ELISA developed in this study is useful screening tool to detect antibodies against CSFV for the large-scale monitoring of classical swine fever.

Serum samples were collected from 938 pigs of 24 farms in Hokkaido, Kagoshima, and Okinawa prefectures in Japan in 2001-2005. Enzyme-linked immunosorbent assay (ELISA) was used for the detection of antibodies to LipL32 antigen which is common to Leptospira interrogans. Samples positive in ELISA were then investigated by microscopic agglutination test for the identification of causal leptospires. Antibodies specific to leptospires of serovars Copenhageni, Bratislava, Australis and Javanica were detected in serum samples of pigs from each of the three districts. In addition, antibodies to leptospires of serovars Autumnalis and Tarassovi were predominantly detected in those from Kagoshima. The present study, thus, revealed that leptospires belonging to different serovars prevail in the pig population in Japan. In addition, it is the first detection of antibodies to leptospires belonging to serovars Javanica and Tarassovi in pigs in Japan.

To examine the development of the tendon pulley of the obturator internus muscle (OI), we observed paraffin sections of 26 human embryos and fetuses (∼6-15 weeks of gestation). The OI was characterized by early maturation of the proximal tendon in contrast to the delayed development of the distal tendon. At 6 weeks, the ischium corresponded to a simple round mass similar to the tuberosity in adults. At 8 weeks, before development of the definite lesser notch of the ischium, initial muscle fibers of the OI, running along the antero-posterior axis, converged onto a thick and tight but short tendon running along the left-right axis. Thus, at the beginning of development, the OI muscle belly and tendon met almost at a right angle. At 10 weeks, the OI tendon extended inferiorly along the sciatic nerve, but the distal part remained thin and loose and it was embedded in the gluteus medius tendon. At 15 weeks, in association with the gemellus muscles, the distal OI tendon was established. The mechanically strong sciatic nerve was first likely to catch the OI muscle fibers to provide a temporary insertion. Next, the ischium developing upward seemed to push the tendon to make the turn more acute along the cartilaginous ridge. Finally, the gemellus muscle appeared to provide inferior traction to the OI tendon for separation from the gluteus medius to create the final, independent insertion. Without such guidance, the piriformis tendon first attached to the OI tendon and then merged with the gluteus medius tendon.