Insecticide susceptibilities in populations of two rice planthoppers, Nilaparvata lugens and Sogatella furcifera, immigrating into Japan in the period 2005-2012.

BACKGROUND: The brown planthopper Nilaparvata lugens and the whitebacked planthopper Sogatella furcifera are both important pests on rice throughout Asia. The major cause of recent outbreaks is thought to be the development of insecticide resistance. Thus, the authors monitored insecticide susceptibilities in populations of these two insects immigrating into Japan in the period 2005-2012. Ten insecticides were tested, including members of the organophosphate, carbamate, pyrethroid, neonicotinoid and phenylpyrazole groups. RESULTS: The LD50 values of N. lugens against imidacloprid increased from 2005 (0.7 µg g(-1)) to 2012 (98.5 µg g(-1)). The resistance ratio (LD50 value in 2012/baseline LD50 value in 1992) was 615.5. In contrast, LD50 values of N. lugens against fipronil were <1.0 µg g(-1) up to 2012, suggesting that N. lugens had developed no insecticide resistance to this insecticide. However, S. furcifera exhibited resistance against fipronil up to 2012. Except for the case of malathion, the resistances of N. lugens against members of the organophosphate and carbamate groups were closely similar in the period 2005-2012 to earlier determinations in 1984 and 1985. CONCLUSION: Species-specific insecticide resistance (imidacloprid resistance in N. lugens and fipronil resistance in S. furcifera) is ongoing in populations of the two planthoppers immigrating into Japan.

PCR-RFLP analysis for identification of Tetranychus spider mite species (Acari: Tetranychidae).

A polymerase chain reaction (PCR)-restriction fragment length polymorphism (PCR-restriction fragment-length polymorphism)-based method for species identification was applied to 14 Tetranychus spider mite species, which were dominant species intercepted at Japanese import plant quarantine. We sequenced the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA (rDNA), which included the partial ends of the 18S and 28S ribosomal RNA genes, 5.8S ribosomal RNA gene, and two internal transcribed spacers (ITS1 and ITS2) for 15 populations of the 14 species. We analyzed the recognition sites of four restriction endonucleases, which had been proposed for discrimination of Japanese Tetranychus species, and constructed a scheme for Tetranychus species identification by PCR-restriction fragment-length polymorphism. We then applied the scheme to 245 individuals from 199 populations, most of them were from foreign countries. As a result, all 14 species were correctly identified using PCR-restriction fragment-length polymorphism. This demonstrates the usefulness of the PCR-restriction fragment-length polymorphism method for the worldwide identification of Tetranychus species.

A rare sugar, d-allose, confers resistance to rice bacterial blight with upregulation of defense-related genes in Oryza sativa.

We investigated responses of rice plant to three rare sugars, d-altrose, d-sorbose, and d-allose, due to establishment of mass production methods for these rare sugars. Root growth and shoot growth were significantly inhibited by d-allose but not by the other rare sugars. A large-scale gene expression analysis using a rice microarray revealed that d-allose treatment causes a high upregulation of many defense-related, pathogenesis-related (PR) protein genes in rice. The PR protein genes were not upregulated by other rare sugars. Furthermore, d-allose treatment of rice plants conferred limited resistance of the rice against the pathogen Xanthomonas oryzae pv. oryzae but the other tested sugars did not. These results indicate that d-allose has a growth inhibitory effect but might prove to be a candidate elicitor for reducing disease development in rice.

Role of hydroperoxide lyase in white-backed planthopper (Sogatella furcifera Horváth)-induced resistance to bacterial blight in rice, Oryza sativa L.

A pre-infestation of the white-backed planthopper (WBPH), Sogatella furcifera Horváth, conferred resistance to bacterial blight caused by Xanthomonas oryzae pv. oryzae (Xoo) in rice (Oryza sativa L.) under both laboratory and field conditions. The infestation of another planthopper species, the brown planthopper (BPH) Nilaparvata lugens Stål, did not significantly reduce the incidence of bacterial blight symptoms. A large-scale screening using a rice DNA microarray and quantitative RT-PCR revealed that WBPH infestation caused the upregulation of more defence-related genes than did BPH infestation. Hydroperoxide lyase 2 (OsHPL2), an enzyme for producing C(6) volatiles, was upregulated by WBPH infestation, but not by BPH infestation. One C(6) volatile, (E)-2-hexenal, accumulated in rice after WBPH infestation, but not after BPH infestation. A direct application of (E)-2-hexenal to a liquid culture of Xoo inhibited the growth of the bacterium. Furthermore, a vapour treatment of rice plants with (E)-2-hexenal induced resistance to bacterial blight. OsHPL2-overexpressing transgenic rice plants exhibited increased resistance to bacterial blight. Based on these data, we conclude that OsHPL2 and its derived (E)-2-hexenal play some role in WBPH-induced resistance in rice.

Species-specific insecticide resistance to imidacloprid and fipronil in the rice planthoppers Nilaparvata lugens and Sogatella furcifera in East and South-east Asia.

BACKGROUND: In 2003 the development of insecticide resistance against neonicotinoids in the brown planthopper (BPH), Nilaparvata lugens (Stål) (Homoptera: Delphacidae), was first observed in Thailand and has since been found in other Asian countries such as Vietnam, China and Japan. However, the LD(50) values of BPH and the whitebacked planthopper (WBPH), Sogatella furcifera (Horváth), against both neonicotinoid and phenylpyrazole insecticides have been poorly reported in many Asian countries. RESULTS: The topical LD(50) values for imidacloprid in the BPH populations collected from East Asia (Japan, China, Taiwan) and Vietnam in 2006 were 4.3-24.2 microg g(-1) and were significantly higher than those collected from the Philippines (0.18-0.35 microg g(-1)). The BPH populations indicated a positive cross-resistance between imidacloprid and thiamethoxam. Almost all the WBPH populations from Japan, Taiwan, China, Vietnam and the Philippines had extremely large LD(50) values (19.7-239 microg g(-1) or more) for fipronil, except for several populations from the Philippines and China. CONCLUSION: Species-specific changes in insecticide susceptibility were found in Asian rice planthoppers (i.e. BPH for imidacloprid and WBPH for fipronil). Insecticide resistance in BPH against imidacloprid occurred in East Asia and Indochina, but not in the Philippines. In contrast, insecticide resistance in WBPH against fipronil occurred widely in East and South-east Asia.

Penetration of ravuconazole, a new triazole antifungal, into rat tissues.

Ravuconazole (BMS 207147, ER-30346) is a long-lasting triazole antifungal agent active against a broad spectrum of fungal pathogens including non-albicans Candida, Aspergillus, Cryptococcus and key dermatophytic fungi. The penetration of ravuconazole into rat tissues was examined. Fifty-five 7-week-old specific pathogen free female rats were used in this study. Plasma, lung and uterus tissue of rats were taken at 1, 2, 4, 8, 12, 16, 24, 32, 48, 60, and 72 h (n = 5) after oral administration of 10 mg/kg of ravuconazole. The quantitative assays of ravuconazole by HPLC after the extraction with diethylether were conducted for each tissue sample homogenate. tmax, t1/2, and Cmax of ravuconazole is 8 h, 16.9 h and 1.68 microg/ml, respectively. The concentrations of ravuconazole in rat uterus and lung tissues were 2-to 6 times higher than the corresponding blood concentrations. The ratio of plasma to lung levels of ravuconazole was superior to the published data of other azoles. Considering its antifungal spectrum, ravuconazole would thus be a good candidate for treatment of deep-seated fungal infections caused by Candida, Aspergillus and Cryptococcus.

A new method for preparing electron microscopic specimens of Helicobacter pylori.

Abstract A new method for preparing electron microscopic specimens of Helicobacter pylori was developed and used to examine the ultrastructure of this bacterium. We have also investigated the morphological changes in the bacterium when exposed to amoxicillin using our new method. Bacterial specimens for electron microscopy are usually prepared by collecting the bacteria by centrifugation during the fixation and dehydration processes. In our new method the bacteria are filtered through and adsorbed onto a filter before fixation, and the entire filter containing the adhered bacteria is fixed and dehydrated. Using this method we were able to obtain electron photomicrographs in which the external appearances or internal structures of the bacteria were well conserved. The advantages of this method are that it uses only a small amount of bacterial suspension, shortens the time required for the dehydration procedure, and keeps the artifacts to the minimum. Amoxicillin treatment resulted in coccoid form with blebs in the bacterial surface and the appearance of vacuoles, granules, and an area of low electron density in the cytoplasm at one and four minimum inhibitory concentrations. These changes were consistent with results previously reported in the literature.