Prevalence of zoonotic Bartonella species among rodents and shrews in Thailand

We investigated the prevalence of Bartonella species in 10 rodent and one shrew species in Thailand. From February 2008 to May 2010, a total of 375 small animals were captured in 9 provinces in Thailand. Bartonella strains were isolated from 57 rodents (54 from Rattus species and 3 from Bandicota indica) and one shrew (Suncus murinus) in 7 of the 9 provinces, and identified to the species level. Sequence analysis of the citrate synthase and RNA polymerase β subunit genes identified the 58 isolates from each Bartonella-positive animal as B. tribocorum in 27 (46.6%) animals, B. rattimassiliensis in 17 (29.3%) animals, B. elizabethae in 10 (17.2%) animals and B. queenslandensis in 4 (6.9%) animals. R. norvegicus, R. rattus, and Suncus murinus carried B. elizabethae, which causes endocarditis in humans. The prevalence of Bartonella bacteremic animals by province was 42.9% of the animals collected in Phang Nga, 26.8% in Chiang Rai, 20.4% in Sa Kaeo, 16.7% in Nakhon Si Thammarat, 12.0% in Surat Thani, 9.1% in Mae Hong Son and Loei Provinces.     

CEP peptide induces susceptibility of Arabidopsis thaliana to non-adapted pathogens

CEP peptide was synthesized and tested for induction of disease susceptibility using Arabidopsis Col-0. When Colletotrichum tropicale was used as a non-adapted fungal pathogen, the conidia germinated to form hyphal-like structures, which successfully penetrated epidermis, eventually causing disease symptoms. In such case, PEN2-, but not PEN3-dependent resistance was likely suppressed by CEP peptide. Similarly, the CEP peptide-mediated disease susceptibility was also effective to a non-adapted bacterial pathogen. Notably, such induced susceptibility was also evident on Arabidopsis mutants lacking the previously identified receptors, suggesting that the CEP peptide modulates Arabidopsis immunity through an unidentified receptor(s).

     

Identification and Biochemical Characterization of Halisulfate 3 and Suvanine as Novel Inhibitors of Hepatitis C Virus NS3 Helicase from a Marine Sponge

Hepatitis C virus (HCV) is an important etiological agent that is responsible for the development of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. HCV nonstructural protein 3 (NS3) helicase is a possible target for novel drug development due to its essential role in viral replication. In this study, we identified halisulfate 3 (hal3) and suvanine as novel NS3 helicase inhibitors, with IC₅₀ values of 4 and 3 µM, respectively, from a marine sponge by screening extracts of marine organisms. Both hal3 and suvanine inhibited the ATPase, RNA binding, and serine protease activities of NS3 helicase with IC₅₀ values of 8, 8, and 14 µM, and 7, 3, and 34 µM, respectively. However, the dengue virus (DENV) NS3 helicase, which shares a catalytic core (consisting mainly of ATPase and RNA binding sites) with HCV NS3 helicase, was not inhibited by hal3 and suvanine, even at concentrations of 100 µM. Therefore, we conclude that hal3 and suvanine specifically inhibit HCV NS3 helicase via an interaction with an allosteric site in NS3 rather than binding to the catalytic core. This led to the inhibition of all NS3 activities, presumably by inducing conformational changes.     

Seroprevalence of Toxoplasma gondii antibodies in stray cats and dogs in the Bangkok metropolitan area, Thailand

Cats and dogs are the most popular pet animals worldwide. Cats are the natural reservoir of Toxoplasma gondii and excrete the resistant oocyst to environments. On the other hands, dogs play a role in the mechanical transmission of the parasite. Stray cats and dogs in the Bangkok metropolitan area are becoming a public concern because there is a considerable increase in their number annually. These facts indicate the risk of mechanically spreading zoonoses including toxoplasmosis to humans since human acquire the infection from infected mammals, either directly or indirectly. In the present study, the presence of T. gondii antibodies was examined in 592 cats and 427 dogs from October 2001 to September 2002 by using a latex agglutination test. T. gondii antibodies were detected in 65 (11.0%) of the 592 cats and 40 (9.4%) of the 427 dogs. The antibody titers in the positive animals ranged from 1:64 to 1:2048. Seroprevalence was significantly higher in female cats than in male cats. The present study suggested that T. gondii was widespread in the stray animals in the Bangkok metropolitan area; therefore, it is essential to control the number of stray cats and dogs in order to reduce the transmission of toxoplasmosis to animals and humans.     

Quantification of genetically modified soybean by quenching probe polymerase chain reaction

Quenching probe (QProbe) polymerase chain reaction (PCR) is a simple and cost-effective real-time PCR assay in comparison with other real-time PCR assays such as the TaqMan assay. We used QProbe-PCR to quantify genetically modified (GM) soybean (Roundup Ready soybean). We designed event-specific QProbes for Le1 (soy endogenous gene) and RRS (recombinant gene), and we quantified certified reference materials containing 0.1, 0.5, 1, 2, and 5% GM soybean. The TaqMan assay was also applied to the same samples, and the results were compared. The accuracy of QProbe-PCR was similar to that of TaqMan assay. When GM soybean content was 0.5% or more, the relative standard deviations of QProbe-PCR were less than 20%. QProbe-PCR is sensitive enough to monitor labeling systems and has acceptable levels of accuracy and precision.     

An atypical bHLH protein encoded by POSITIVE REGULATOR OF GRAIN LENGTH 2 is involved in controlling grain length and weight of rice through interaction with a typical bHLH protein APG

Grain size is an important yield component in rice, however, genes controlling the trait remain poorly understood. Previously, we have shown that an antagonistic pair of basic helix-loop-helix (bHLH) proteins, POSITIVE REGULATOR OF GRAIN LENGTH 1 (PGL1) and ANTAGONIST OF PGL1 (APG), is involved in controlling rice grain length. Here, we report the involvement of another atypical bHLH protein gene, POSITIVE REGULATOR OF GRAIN LENGTH 2 (PGL2), in the regulation of rice grain length. Over-expression of PGL2 in the lemma/palea increased grain length and weight in correlation with the level of transgene expression. Observation of the inner epidermal cells of lemma of PGL2-overexpressing lines revealed that the long grain size is caused by an increase in cell length. PGL2 interacts with a typical bHLH protein APG, a negative regulator of rice grain length and weight, in vitro and in vivo. It was reported that overexpression of BU1 (BRASSINOSTEROID UPREGULATED 1), the closest homolog of PGL2, caused an increase in grain length. However, we detected no interaction between BU1 and APG. These findings suggest that PGL2 and PGL1 redundantly suppress the function of APG by forming heterodimers to positively regulate the rice grain length, while the pathway through which BU1, the closest homolog of PGL2, controls grain length is independent of APG.     

Characterization of the suppressive effects of the biological control strain VAR03-1 of <Emphasis Type="Italic">Rhizobium vitis</Emphasis> on the virulence of tumorigenic <Emphasis Type="Italic">R. vitis</Emphasis>

Rhizobium vitis: strain VAR03-1 is a biological control agent that suppresses grapevine crown gall disease caused by a tumorigenic strain of R. vitis (Ti). Both acetosyringone-induced expression of a virulence gene and the growth of Ti were suppressed in vitro when it was cultivated in the VAR03-1 culture filtrate. These inhibitory effects were reduced by high-temperature treatment or incubation for 72 h. Both activities were detected in the high molecular weight fraction (>?100 kDa) of the filtrate. Our results suggest that the antagonistic effects of VAR03-1 on Ti are mediated by large particle(s) released in the culture media.     

Serodiagnosis of Babesia gibsoni infection in dogs by an improved enzyme-linked immunosorbent assay with recombinant truncated P50

The surface antigen P50 of Babesia gibsoni is an important candidate for the development of a diagnostic reagent for canine piroplasmosis. In order to establish an effective diagnostic method for practical use, the gene encoding truncated P50 (P50t) lacking a signal peptide and C-terminal hydrophobic regions were cloned and expressed in Escherichia coli as a fusion protein with glutathione S-transferase (GST). More than 90% portion of the GST-P50t was expressed as a soluble form, in contrast with GST-P50f (full-length), which was completely expressed as an insoluble form. This result indicates that removal of the hydrophobic signal peptide and C-terminus had dramatically improved its hydrophilicity. The purified GST-P50t was tested in an enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to B. gibsoni in dogs. The ELISA with GST-P50t clearly differentiated between B. gibsoni-infected dog sera and uninfected dog sera. In addition, the ELISA detected no cross-reactivity with sera from dogs experimentally infected with the closely related parasites, B. canis canis, B. canis vogeli, and B. canis rossi. Field serum samples collected from dogs in Japan and China were examined for the diagnosis of B. gibsoni infection by using the ELISA. 14.5% (9/62), 5.8% (7/120), and 5.4% (2/37) of tested samples were positive for dogs from Okinawa, Yamaguchi, and Osaka prefectures, Japan, respectively. On the other hand, 4.8% (2/41) of tested samples were positive for dogs from Nanjing, China. These results suggest that the GST-P50t could be a reliable reagent for practical use in ELISA for the serodiagnosis of canine piroplasmosis caused by B. gibsoni.     

Serological evidence of infection of Anaplasma and Ehrlichia in domestic animals in Xinjiang Uygur Autonomous Region area, China

Serological methods were utilized to detect Anaplasma and Ehrlichia infection in domestic animals in Xinjiang Uygur Autonomous Region, China. By using an indirect immunofluorescence assay (IFA), antibodies that reacted with Anaplasmaphagocytophilum and Ehrlichiachaffeensis were detected mainly in ruminants kept on pastureland in Altai, Ili and Kashgar area. Antibody titers up to 1:320 were recorded. These results indicate that ruminants kept in these areas may be infected with some species of Anaplasma and Ehrlichia.     

The forming process of the peculiar Mn-mottling in the polder

In the subsurface horizons of some paddy fields at the Kojima polder, Okayama Prefecture, the authors noticed a peculiar Mn-mottling, which had not been found in the common paddy fields. Several paddy soils in varying stages of maturity were carefuIIy surveyed at the Kojima polder. The soils were tentatively named OK4, OK7, OKS, and OK9. The ages of the fields after reclamation were about 10, 100, 150, and 250 years, respectively. Undisturbed samples were collected from 3 soils (OK4, OK7, and OKS) for microscopic observation and chemical analysis.