RNA 3 deletion mutants of beet necrotic yellow vein virus do not cause rhizomania disease in sugar beets

ABSTRACT Two mutant strains of beet necrotic yellow vein virus (BNYVV) containing deletions in RNA 3 were obtained by single lesion transfers in Tetragonia expansa. The deleted regions encode either 94 or 121 amino acids toward the C-terminal part of the 25-kDa protein (P25). Wild-type and mutant virus strains were inoculated by Polymyxa betae to sugar beet seedlings of susceptible and partially resistant cultivars. No differences were found in virus content in rootlets between mutant and wild-type viruses or between susceptible and resistant cultivars after culture for 4 weeks in a growth cabinet. However, when virus-inoculated seedlings were grown in the field for 5 months, the wild-type virus caused typical rhizomania root symptoms (69 to 96% yield loss) in susceptible cultivars, but no symptoms (23% loss) developed in most plants of the resistant cultivar, and BNYVV concentrations in the roots were 10 to 20x lower in these plants than in susceptible plants. In contrast, the mutant strains caused no symptoms in susceptible or resistant cultivars, and the virus content of roots was much lower in both cultivars than in wild-type virus infections. Wild-type RNA 3 was not detectable in most of the taproots of a resistant cultivar without any symptoms, suggesting that replication of undeleted RNA 3 was inhibited. These results indicate that the P25 of BNYVV RNA 3 is essential for the development of rhizomania symptoms in susceptible cultivars and suggest that it may fail to facilitate virus translocation from rootlets to taproots in the partially resistant cultivar.     

The discovery of pyridalyl: a novel insecticidal agent for controlling lepidopterous pests

Synthesis of analogues of two compounds with known insecticidal activity, both of which contain a 3,3-dichloro-2-propenyloxy group, produced 2-(trifluoromethyl)-4-phenoxyphenyl 3,3-dichloro-2-propenyl ether, which had weak activity against lepidopterous larvae. Structural modifications around this lead compound led to the development of pyridalyl [Pleo, S-1812; 2,6-dichloro-4-(3,3-dichloroallyloxy)phenyl 3-[5-(trifluoromethyl)-2-pyridyloxy]propyl ether], which belongs to a new class of insecticides. Pyridalyl gives very good control of various lepidopterous and thysanopterous pests on cotton and vegetables, without phytotoxicity. It controls populations of Heliothis virescens F and Plutella xylostella (L) which are resistant to various currently used insecticides. It also produces unique insecticidal symptoms, so it may have a different mode of action from other existing insecticides. Pyridalyl is also less harmful than existing insecticides to various beneficial arthropods, so it should provide an important tool in IPM and insecticidal management programmes for the control of lepidopterous and thysanopterous pests. The first market introduction is expected in Japan and some Asian countries in the years between 2004 and 2005.     

Analysis of antibody response by temperature-sensitive measles vaccine strain in the cotton rat model

Measles virus (MeV) vaccine strain, AIK-C, is temperature sensitive (ts), which is thought to be associated with attenuation of virus pathogenicity. In this study, replication and antibody response were examined in cotton rats using viruses carrying different forms of the P gene, which is responsible for the ts phenotype of strain AIK-C and its parental Edmonston strain. When cotton rats were inoculated intranasally, ts viruses neither replicated in lungs, nor reproducibly generated an antibody response. When inoculated intramusculary (i.m.), however, ts strains raised an antibody titer in all animals. This response was not observed when ultraviolet-inactivated virus was used. ts virus, inoculated i.m., was recovered from cotton rat drainage lymph nodes. These results suggest that ts virus, inoculated i.m., could replicate in the cotton rat, presumably at the superficial lymph node, and induce an antibody response. Therefore, cotton rats can serve as a small-animal model for investigating immune responses to safer ts vaccine, as well as recombinant vaccine using AIK-C as a vector for protection against other infectious agents.     

Serological evidence of influenza A virus infection in Kuril harbor seals (Phoca vitulina stejnegeri) of Hokkaido, Japan

For proper management and conservation of the Kuril harbor seal (Phoca vitulina stejnegeri) through disease control, serological analysis was performed for influenza A virus infection in free-ranging seals in Hokkaido, Japan. Serum samples were collected from seals at Nosappu (231 seals), Akkeshi (16) and Erimo (75), between 1998 and 2005, and were analyzed by ELISA. Antibodies to the influenza A virus were detected only in seals from Nosappu. The incidences were 11% (1/9), 3% (2/66), 12% (7/59) and 6% (5/77) in 1998, 2003, 2004 and 2005, respectively. These suggest sporadic infection. Because antibody-positive seals included juvenile seals in each year, the infections were considered to have been circulated since no later than the late 1990s until recent years. ELISA-positive sera were analyzed by hemagglutination inhibition (HI) tests to determine the subtypes. Antibodies to the H3 and H6 subtypes were detected in 10 and 2 sera, respectively. Two of the sera that had antibodies to the H6 subtype also had antibodies to the H3 subtype. These two seals were considered to have been infected with both the H3 and H6 subtypes. This is the first investigation to find antibodies to the H6 subtype in seals. Although the H6 subtype had been isolated only from avians, genetic analysis had suggested that the H6 subtype could become a novel mammalian pathogen. For definitive diagnosis, detection of the virus from the tissue or mucus of seals is required.     

The mechanism of bentazon selectivity

Absorption, translocation and metabolism of [¹⁴C]3-isopropyl-2,1,3-benzothiadiazin-4-one-2,2-dioxide (bentazon) by several plant species were investigated to determine the mechanism of bentazon selectivity.Marked selective phytotoxicities were observed between resistant rice (Oryza sativa L.) and susceptible Cyperus serotinus Rottb. when treated with bentazon. Absorption and transolcation of bentazon did not differ greatly between highly resistant rice and susceptible C. serotinus. However, a marked difference in bentazon metabolism occurred between the two species. In rice about 80% of the absorbed bentazon was metabolized within 24 h, and after 7 days about 85% was converted to a major water-soluble metabolite and unchanged bentazon was only 5%. In C. serotinus 50–75% of the radioactivity was unchanged bentazon after 7 days.Large amounts of water-soluble metabolites were detected in root-treated resistant plants such as barnyardgrass (Echinochloa crus-galli Beauv.), soybean (Glycine max Merr.) and corn (Zea mays L.), but only small amounts were present in such susceptible plants as Sagittaria pygmaea Miq. and Eleocharis kuroguwai Ohwi. Therefore, the mechanism of bentazon selectivity appears to be a difference between resistant and susceptible species in their ability to metabolize and detoxify bentazon.The major metabolite in rice was identified as 6-(3-isopropyl-2,1,3-benzothiadiazin-4-one-2,2-dioxide)-O-β-glucopyranoside, determined by GC-MS, NMR, IR and gas chromatography after hydrolysis with sulfuric acid or β-glucosidase.     

Identification of low-molecular weight glutenin subunits of wheat associated with bread-making quality

Although it is known that the compositions of low‐molecular weight glutenin subunits (LMW‐GSs) are important factors for bread‐making quality of wheat, it is still not clear which LMW‐GSs confer improved bread‐making quality and how those LMW‐GSs interact with high‐molecular weight (HMW) GSs. Using a hard red winter wheat line with good bread‐making quality and a Japanese wheat cultivar with poor quality as well as their progeny we identified LMW‐GSs associated with the bread‐making quality. One such LMW‐GS, KS2, which had a molecular weight of 42 kDa and was allelic to HS1, was associated with bread‐making quality. Furthermore, by using four recombinant inbred lines with different HMW‐GS and LMW‐GS combinations, KS2 and HMW‐GS 5+10 showed interaction effects on the bread‐making quality. Two‐dimensional polyacrylamide gel electrophoresis (Page) analysis showed that KS2 consists of two protein components and that HS1 is composed of three components. The N‐terminal amino acid sequences of the five components were identical to the most frequently analysed sequence of LMW glutenin components.     

Genetic characterization and susceptibility on poultry and mammal of H7N6 subtype avian influenza virus isolated in Japan in 2009

From February to March 2009, six strains of H7N6 subtype avian influenza virus were isolated from quails in three farms in Aichi prefecture in Japan. The isolates were shown to be low pathogenic for chicken by the examination performed using the "Manual of Standards for Diagnostic Tests and Vaccines" by World organisation for Animal Health (OIE). The deduced amino acid sequence at the cleavage site was PE (I/Q/L) PKRR (nucleotide sequences were cct gaa (a/c) (t/a) a cc (a/g) aaa aga aga), suggesting persistence in domestic poultry for some time. The direct putative ancestor strain could not be elucidated by phylogenetic analysis of all genome segments of the quail isolates. Diverged date from a putative common ancestor in a non-rooted phylogenetic tree among quail viruses was estimated between March 2002 and July 2004. Three putative N-linked glycosylation sites resided in the vicinity of the receptor binding pocket of HA1 region. They are considered to decrease the reactivity of neutralizing antibody against the virus. Experiments for the infectivity and pathogenicity of a quail strain to poultry indicated that the quail isolate had higher infectivity to quails than chickens and ducks. Direct and dust-borne and/or droplet-borne transmissions among quail were proven in quails with and without direct contact with experimentally infected quails. The virus is seldom transmitted among chickens either directly or indirectly, and indirect transmission from infected quails to chickens was not observed. The pathogenicity of the quail strain for mammalian, pig and mouse was low, although it could replicate in those animals.     

Distribution of the Steller sea lion Eumetopias jubatus during winter in the northern Sea of Japan, along the west coast of Hokkaido, based on aerial and land sighting surveys

Aerial and land-based sighting surveys were conducted to clarify the distribution of Steller sea lions during winter in the northern Sea of Japan, along the west coast of Hokkaido, from 2001 to 2003. Aerial surveys revealed that sea lions gathered around the Rishiri-Rebun Islands in March 2002 and 2003, and between Iwanai and Cape Obana in February 2003. Higher numbers of sea lions were also confirmed at Cape Ofuyu and Cape Kamui on the central-west coast compared to that at the haul-out sites in the 1980s on the upper-west coast in March 2002 and 2003. Additionally, fisherman observed sea lions along the coast of the Tsugaru Strait from February to May 2003, where the presence of sea lions was not reported in the 1980s. These facts suggest that sea lions have recently expanded their distribution southward. Land-based surveys at the recent haul-out sites indicated that sea lions in the central-west coast were composed mainly of adult and subadult males (average: Cape Ofuyu 75.2%, Cape Kamui 69.5%), which stayed from November 2002 to May 2003. In this area, sea lions arrived earlier and stayed longer than in the 1980s.     

Ecological studies of Yersinia enterocolitica I. Dissemination of Y. enterocolitica in pigs

Pigs were examined on five farms for carrier status of Yersinia enterocolitica. Yersinia enterocolitica biovar 4, serovar 3, phagovar VIII was isolated consistently from the feces of fattening pigs on one farm and sporadically from those of similar pigs on the other farms and of sows on all five farms, during a one-year period of weekly surveys. Seasonal variation was not a feature of fattening pigs on a highly contaminated farm. In other pigs, however, the organisms were not isolated during the summer months. On a highly contaminated farm, the organisms were excreted in the feces of 8 to 15-week-old pigs within 1–3 weeks of entering pens which were thought to be contaminated with the organisms. On a detailed observation of natural infection of Y. enterocolitica in eight pigs, the organism appeared in the pigs' feces within 2–7 weeks of them being moved to a pen which had been washed thoroughly after becoming contaminated, by a previous group of pigs, with feces containing 10⁵ viable organisms per g. Thus, Y. enterocolitica is apparently transmitted from infected feces or picked up from the floor of a contaminated pen, and the regular schedule of pig movement among the pens is an important factor in the spread of Y. enterocolitica within a piggery. Intestinal colonization continues for a long time and does not occur by re-infection. The organisms were not isolated from eight pigs at the time of slaughter, and their serum O-agglutinin titers were $\text{1}\text{40}$ or less. Thus, circulating antibody may not inhibit intestinal colonization by Y. enterocolitica.     

Therapeutic effects of Toxoplasma lysate antigen on 20-methylcholanthrene-induced BALB/c mouse tumors.

Therapeutic effects of Toxoplasma lysate antigen (TLA) were studied in mice bearing the tumor in the second passage of 20-methylcholanthrene (MC)-induced tumor cells. Intramuscular administration of TLA 7 days after the tumor-cell inoculation caused apparent inhibition of the tumor growth on day 14. The second treatment facilitated the therapeutic effects. Intravenous transfer of spleen cells prepared from TLA-sensitized mice into tumor-bearing mice also represented the growth inhibitory effects. Prominent effects were seen when the transferred cells were prepared 5 days after sensitization of donor animals. The inhibitory effects were absent in the groups transferred only the adherent cells or the non-adherent cells prepared from sensitized mice. The strongest inhibitory effect was observed in the group to which both adherent and non-adherent spleen cells were transferred simultaneously from sensitized mice. In in vitro experiments, spleen cells obtained from sensitized mice showed cytolytic effect on P-815 or YAC-1 cells after the secondary stimulation in vitro with TLA. Large non-adherent cells containing densely packed granules were induced when cultured with the adherent cells obtained from sensitized mice. These results revealed that TLA can inhibit the growth of the chemically-induced transplantable tumors by activation of adherent and non-adherent spleen cells.