Prevalence of tick-borne Rickettsia and Ehrlichia in Ixodes persulcatus and Ixodes ovatus in Tokachi district, Eastern Hokkaido, Japan

DNA from 111 ticks collected by flagging in Tokachi district, Eastern Hokkaido, Japan were examined for infection with Rickettsia and Ehrlichia, by PCR and sequencing methodology. For Rickettsia, analysis of the partial sequence of the citrate synthase gene was successfully performed on 11 DNA samples from I. persulcatus, and 7 of them showed 99.8% identical with Rickettsia helvetica while the other 4 showed 99.8% identical with ;Candidatus Rickettsia tarasevichiae'. For Ehrlichia, a partial sequence of the 16S rRNA gene detected from I. persulcatus was 100% identical with that from Ehrlichia muris, and another DNA sample from I. ovatus showed 99.8% identical with Ehrlichia species detected from I. ovatus. The results suggest that the pathogens detected here might be distributed in this area.     

Characterization of CTLA-4, PD-1 and PDL-1 of swamp and riverine type water buffaloes

Characterization of CTLA-4, PD-1 and PDL-1 genes from swamp and riverine type water buffaloes was done by molecular cloning, sequencing and phylogenetic analysis. The cloned cDNA of CTLA-4, PD-1 and PDL-1 contained an open reading frame of 666, 849 and 870 nucleotides, encoding a polypeptide of 221, 282 and 298 amino acids, respectively. Nucleotide sequence homology of both CTLA-4 and PDL-1 had 99.8% in swamp and riverine type, which gives the identical polypeptide. Meanwhile, PD-1 genes of swamp and riverine type water buffaloes had 99.2% of homology in nucleotide sequence, which has substitution of two amino acid residues. The hexapeptide motif, phosphatidylinositol 3′-kinase and potential glycosylation sites were conserved within the tribe Bovinae. Phylogenetic analysis confirmed the degree of relationship between the bubaline species and justify the distinctness of each breeds by the bootstrap value generated.     

Comparison of the characters of the plaque-purified viruses from foot-and-mouth disease virus O/JPN/2000

At least two biotypes were observed at the 2nd passage stage after the isolation of Foot-and-mouth disease Virus (FMDV) O/JPN/2000 strain. These 2 types of viruses differed from their plaque phenotypes and were distinguishable by using a monoclonal antibody (MAb) 64G8 that was made for the FMDV O/JPN/2000 strain. One of these 2 biotypes formed small plaque (SP) and with immuno staining showed a positive reaction to MAb 64G8, while the other formed clear large plaque (LP) and did not react with MAb 64G8. The amino acid sequences of the capsid coding region (VP1-VP4) of the SP virus (SPV) and the LP virus (LPV) revealed two substitutions on the 133rd amino acid in VP2, and the 56th amino acid in VP3. These amino acid changes of SPV and LPV are Asn to Asp, Arg to His, respectively. The Arg of the 56th amino acid in VP3 that have been known as critical position of cell culture adapted virus. Only LPV showed high pathogenicity in suckling mice, and its LD(50) was calculated to be about 10(2) TCID(50)/0.1 ml. These results showed that the SPV that existed at the 2nd passage stage from isolation was a low virulence virus, which may suggest why the pathogenicity of O/JPN/2000 did not show clear symptoms in infected cattle.     

The detection of the meq gene in chicken infected with Marek's disease virus serotype 1

In the genome of strains of very virulent Marek's disease virus serotype 1(vvMDV1), such as Md5 and RB1B, the meq open reading frame (ORF) encoding a 339-amino-acid bZIP protein, is present, while a slightly longer meq ORF, termed as L-meq, in which a 180-bp sequence is inserted into the meq ORF is found in other strains of MDV1, such as CV1988/R6 and attenuated JM. When chickens were infected with vvMDV1 strains and the meq gene was amplified by nested polymerase chain reaction (PCR), the meq gene was detected throughout the experimental period for 7 weeks post inoculation (pi). However, the L-meq gene was also detected at 3 to 5 weeks and 3 to 4 weeks pi. in Md5-infected and RB1B-infected chickens, respectively. In the case of chickens infected with an attenuated MDV1, the JM strain, the L-meq gene was detected at 2 to 7 weeks pi., and the meq gene was also detected at 2 to 6 weeks pi. Both L-meq and meq genes were detected in chickens infected with an attenuated nononcogenic vaccine strain of MDV1 (CVI988/R6), throughout the experimental period. Though quantitative PCR was not performed, a larger amount of the PCR products corresponding to the L-meq than the meq gene was amplified from chickens infected with JM or CVI988/R6. These results suggest that a dynamic population shift between the MDV subpopulations displaying meq and L-meq genes occurs in chickens during the course of MDV infection. Since the MDV subpopulation that displays the L-meq gene only displays it during the latent phase, the L-meq and its gene product, if any, might contribute to the maintenance of the MDV latency.     

Does the mutualism between wood ants (Formica rufa group) and Cinara aphids affect Norway spruce growth?

Ant–aphid mutualisms, in which ants tend aphids, which in turn provide honeydew to the ants, are widespread and have been shown to affect plant growth. In boreal forests the effect of ant–aphid mutualism on tree growth can vary with stand age, because forest clear-cutting harms the ecologically most dominant ant partner in such mutualisms, wood ants (Formica rufa group). We studied whether the mutualism between wood ants and Cinara aphids affects the growth of boreal Norway spruces (Picea abies L. Karst.) in stands of different ages. In boreal forests, conifers, unlike deciduous trees, have only few defoliating insects, and therefore we expected the growth loss of conifers due to sap sucking by aphids not to be compensated by reduced insect herbivory due to predation by wood ants. The study was conducted in medium-fertile spruce-dominated stands in eastern Finland. We used stands of four different age classes (5, 30, 60 and 100 years) and selected ten spruces heavily visited and ten spruces lightly visited by ants around five medium-sized ant mounds in each stand age class. The access of ants was blocked on half of the trees in both groups. In the 5-year-old stands, the mean annual height growth of individual heavily visited seedlings was 16.3% greater than in the ones where ant traffic was blocked, but this difference was not significant. In the 30-year-old stands, the mean annual radial growth of the heavily visited spruces was 7.3% smaller than in trees where ant traffic was blocked, and this difference was significant. The mutualism had no significant effect on the radial growth in the 60- and 100-year-old stands. In the 60-year-old stands, however, the spruces that were visited heavily prior to the beginning of the study grew significantly less relative to their past growth than the initially lightly visited trees during the study. This suggests that the ant–aphid mutualism may have long-term effects on tree growth. The ant–aphid mutualism had no significant effect on the growth at the stand level. The results indicate that ant–aphid mutualism can have a significant effect on the growth of individual spruces, but the effect is negligible at ecosystem level.     

Antimicrobial activity against <Emphasis Type="Italic">Streptococcus sobrinus</Emphasis> and glucosyltransferase inhibitory activity of taxifolin and some flavanonol rhamnosides from kempas (<Emphasis Type="Italic">Koompassia malaccensis</Emphasis>) extracts

Twenty plant materials collected from the islands of Java and Kalimantan in Indonesia were extracted with 50% aqueous ethanol (crude extract). The crude extracts were assayed for antimicrobial activities against Streptococcus sobrinus and for glucosyltransferase (GTase) inhibition. Fourteen extracts inhibited the growth of S. sobrinus by more than 50% and six extracts inhibited GTase activity by more than 50% at a concentration of 100 μg/ml. Koompassia malaccensis (kempas) extracts showed 90% depression of S. sobrinus growth and 80% inhibition of GTase activity at a concentration of 100 μg/ml. Kempas crude extracts were subjected to column chromatography using Sephadex LH-20 and then preparative high-performance liquid chromatography to isolate four compounds A, B, C, and D. These compounds were identified as taxifolin and the flavanonol rhamnoside isomers neoastilbin, astilbin, and isoastilbin, respectively, from ¹H and ¹³C nuclear magnetic resonance (NMR) spectra and other two-dimensional NMR techniques (COSY, HMBC, and HMQC). Each compound depressed the growth of S. sobrinus over a concentration range of 9.3242.7 μg/ml and showed GTase inhibitory activity with IC₅₀ values in the range 27.4–57.3 μg/ml. Taxifolin and flavanonol rhamnoside isomers isolated for the first time from kempas could be potent compounds for preventing dental caries. Part of this report was presented at the 57th Annual Meeting of the Japan Wood Research Society Conference, Hiroshima, 2007     

Bovine leukaemia virus envelope peptides cause immunomodulation in BALB/c mice

Immunomodulatory activity of two bovine leukaemia virus envelope (BLVEnv) derived peptides were examined in BALB/c mice. One is peptide homologous to CKS-17 which is known as a 17-amino acid peptide derived from p15E of feline leukaemia virus (CKS-17/BLV), and the other is an 18-amino acid synthetic peptide of BLV Env 61-78 (pep61). Priming with CKS-17/BLV in vitro, as well as CKS-17, significantly suppressed the mitogen-induced proliferative responses of spleen cells in naive BALB/c mice. In addition, priming of spleen cells with pep61 in vitro and in vivo resulted in suppression of lipopolysaccaride-induced B-cell proliferative response. This suppression was partially due to the basic amino acid sequence in the peptide because if the pep61-derived peptide lacking Arg was used, this inhibitory activity was partially restored. In contrast, pep61 enhanced both concanavalin A-stimulated proliferative response and IL-2 production. These findings showed that pep61 may contribute to the modification of the host immune responses in the course of BLV infection.     

Sequence analysis of the genes encoding the phosphoprotein of recent isolates of canine distemper virus in Japan

The nucleotide sequences of the phosphoprotein (P) of canine distemper virus (CDV) strains isolated between 1992 and 1996 in Japan were determined. This is the first report of the complete sequences of the P genes of recently prevalent CDV strains. The deduced amino acid sequences of the P, C and V proteins showed that in the new Japanese isolates, these proteins have approximately 93%, 90-91% and 92% identities with those of the Onderstepoort vaccine strain, respectively. The predicted functional regions were conserved. RNA editing resulting in a shift to the open reading frame (ORF) of the V protein was shown to occur with the same efficiency in both the field isolates and vaccine strain.     

Dietary value of benthic diatoms for post-larval abalone <Emphasis Type="Italic">Haliotis diversicolor</Emphasis> associated with feeding transitions

ABSTRACT:   The feeding behavior and growth of post-larval Haliotis diversicolor with initial shell lengths (SL) of approximately 500 μm (Exp. 1-1 and 1-2), 800 μm (Exp. 2), and 1200 μm (Exp. 3) were studied in a laboratory setting while they fed on four species of benthic diatom Achnanthes longipes , Cocconeis sublittoralis , Cylindrotheca closterium , and Navicula ramosissima . Exp. 1-1 and 1-2 revealed no marked differences in post-larval growth rates (mean 24–39 μm SL/day) among the diatom species. However, marked differences in growth rates among the species were revealed in Exp. 2 and 3. Three species, A. longipes , Co. sublittoralis, and Cy. closterium , produced faster growth (Exp. 2 mean 29–51 μm/day, Exp. 3 mean 36–44 μm/day) than N. ramosissima (Exp. 2 mean 18 μm/day, Exp. 3 mean 23 μm/day). Post-larvae fed N. ramosissima had lower digestion efficiency (42.8%) than those fed other diatom species (90.7–100%). Diatom extracellular substances appeared to be principally used from post-settlement to 800 μm SL, and diatom cell contents were required to produce rapid growth of larger post-larvae (>800 μm SL). It is likely that the availability of each diatom for post-larvae was affected by diatom morphology, attachment strength, frustule strength, and post-larval size.