Kazuhiko Namikawa

Artesunate, a potential drug for treatment of Babesia infection.

The effects of artesunate, a water-soluble artemisinin derivative, against Babesia species, including Babesia bovis, Babesia gibsoni and Babesia microti were studied. Cultures of B. bovis and B. gibsoni were treated with 0.26, 2.6, 26 and 260microM artesunate, showing inhibition of parasite growth at concentrations equal to and greater than 2.6microM artesunate by days 3 post-treatment for B. gibsoni and B. bovis in a dose-dependent manner. Consistent with in vitro experiments, artesunate was effective in the treatment of mice infected with B. microti at doses equal to and greater than 10mg/kg of body weight on days 8-10 post-infection. Taken together, these results suggest that artesunate could be a potential drug against Babesia infection.

An Outbreak of Trixacarus caviae Infestation in Guinea Pigs at an Animal Petting Facility and an Evaluation of the Safety and Suitable Dose of Selamectin Treatment

abstract:  In June 2009, 27 guinea pigs kept at an animal petting facility at a zoo in Kanagawa Prefecture, Japan, were observed to scratch intensely, weaken, and develop lesions. Three sarcoptiform mites were found in skin scrapings taken from affected areas of 2 guinea pigs, and they were identified as Trixacarus caviae by morphological examination. This result confirmed the presence of T. caviae in Japan. For treatment, doses of 13.6–18.75 mg/head of selamectin were administered in a topical preparation applied to a single spot on the skin on the back of the neck, and no side effects were observed. In April 2010, a second outbreak of mange occurred at the zoo, and, following investigation, 2 mite eggs were observed. It was, therefore, thought probable that the mites had survived during the winter within nonclinical carriers. Accordingly, doses of 5.0–7.5 mg/head of selamectin were applied on days 0 and 28, after which clinical symptoms disappeared and general condition improved. This dose of selamectin was thus shown to be a suitable and economical treatment for guinea pigs infested with the mites. Because the mite is not always easily observed in infested guinea pigs and the potential for human infestation exists, clinicians should not hesitate to treat when the clinical presentation suggests infestation, particularly in a setting such as an animal petting facility, where large numbers of children and adults have direct contact with the animals.

Characterization of the Babesia gibsoni 12-kDa protein as a potential antigen for the serodiagnosis.

A novel gene, BgP12, encoding a 12-kDa protein was identified from Babesia gibsoni. The full-length cDNA of BgP12 contains an open reading frame of 378 bp, corresponding to 126 amino acid (aa) residues consisting of a putative 26 aa signal peptide and a 100 aa mature protein. The recombinant BgP12 (rBgP12) lacking the N-terminal signal peptide was expressed in Escherichia coli as a soluble glutathione S-transferase (GST) fusion protein (rBgP12) that produced an anti-rBgP12 serum in mice after immunization. Using this anti-rBgP12 serum, a native 12-kDa protein in B. gibsoni was recognized by Western blot analysis. Immunofluorescent antibody tests (IFAT) revealed that BgP12 was mainly seen during the ring stage of B. gibsoni trophozoite. An indirect enzyme-linked immunosorbent assay (ELISA) using the rBgP12 detected specific antibodies in the sequential sera of a dog experimentally infected with B. gibsoni beginning 10 days post-infection to 442 days post-infection, even when the dog became chronically infected and showed a low level of parasitemia. Moreover, the antigen did not show cross-reaction with antibodies to the closely related apicomplexan parasites, indicating that the rBgP12 might be an immunodominant antigen for B. gibsoni infection that could be used as a diagnostic antigen for B. gibsoni infection with high specificity and sensitivity.

Four promising antigens, BgP32, BgP45, BgP47, and BgP50, for serodiagnosis of Babesia gibsoni infection were classified as B. gibsoni merozoite surface protein family

We determined the molecular characteristics of four proteins, BgP32, BgP45, BgP47, and BgP50, of Babesia gibsoni. Localization by subcellular fractionations followed by Western blotting revealed that the corresponding native proteins belong to merozoite surface protein family of B. gibsoni (BgMSP). Moreover, antisera against either rBgP45 or rBgP47 cross-reacted with all the proteins of the BgMSP family on ELISA and IFAT analyses. Of the four candidate antigens, ELISA with rBgP45 yielded high sensitivity, and ELISA with rBgP32 resulted in high specificity and in concordance with IFAT results.

Booster effect of canine distemper, canine parvovirus infection and infectious canine hepatitis combination vaccine in domesticated adult dogs

Domesticated adult dogs with antibody titer classified as below ‘high’ to one or more of canine distemper virus (CDV), canine parvovirus type‐2 (CPV‐2) and canine adenovirus type‐1 (CAdV‐1) were then given an additional inoculation, and the effectiveness of this booster evaluated 2 months later. Consequently, CDV and CAdV‐1 antibody titer experienced a significant increase, but the same effect was not observed in the antibody titer of CPV‐2. These findings suggest that with additional inoculation, a booster effect may be expected in increasing antibody titers for CDV and CAdV‐1, but it is unlikely to give an increase in CPV‐2 antibody titer.

Antibodies to parvovirus, distemper virus and adenovirus conferred to household dogs using commercial combination vaccines containing Leptospira bacterin.

To examine how the inclusion (+) or exclusion (–) of inactivated Leptospira antigens in a vaccine for canine parvovirus type 2 (CPV-2), canine distemper virus (CDV) and canine adenovirus type 2 (CAdV-2) affects antibody titres to CPV-2, CDV and CAdV-1 antigens, household dogs were vaccinated with commercially available vaccines from one of three manufacturers. CPV-2, CDV and CAdV-1 antibody titres were measured 11 to 13 months later and compared within three different age groups and three different bodyweight groups. There were significant differences between CPV-2 antibody titres in dogs vaccinated with (+) vaccine and those vaccinated with (–) vaccine for two products in the two-year-old group and for one product in the greater than seven-year-old group; no significant differences were seen that could be attributed to bodyweight. No differences in CDV antibody titres were observed within age groups, but a significant difference was seen in the 11 to 20 kg weight group for one product. Significant differences in CAdV-1 antibody titres were seen for one product in both the two-year-old group and the ≤10 kg weight group.