Molecular epidemiological analysis of bat rabies viruses in Brazil

A molecular epidemiological analysis was performed in 19 rabies viruses (RVs) isolated from haematophagous, frugivorous and insectivorous bats, in Sao Paulo, Brazil. The authors carried out RT-PCR for amplification of the RV nucleoprotein (N) gene, and determined 1,335 nucleotide sequences of N gene by direct sequencing method. Phylogenetic analysis, which was based on the N gene of Brazilian RV isolates identified presently and previously, revealed that RVs isolated from bats were genetically divided into four lineages had a tendency to depend on the host bat species. The first lineage consisted mainly of haematophagous bat (Desmodus rotundus) isolates, including frugivorous bat (Artibeus spp.) isolates. Other three lineages consisted of insectivorous bat isolates; mainly Eptesicus spp., Molossus spp. and Nyctinomops spp. isolates, respectively. These results indicate a possibility of that there are bat species-specific RV variants in Brazil.     

Effects of inoculum density of pinewood nematode on the development of pine wilt disease in Japanese black pine seedlings pretreated with simulated acid rain

Six‐month‐old Japanese black pine seedlings (Pinus thunbergii) were exposed to simulated acid rain (SAR) at pH 3 and 2 three times a week. After treatment for 2 months, the seedlings were inoculated with a virulent isolate (S10) of the pinewood nematode (Bursaphelenchusxylophilus), at three inoculum levels (P_i = 50, 160 or 500 nematodes per seedling). In seedlings inoculated with 500 nematodes, both population growth of nematodes and disease development were accelerated by pretreatment with SAR at pH 3 or 2. In seedlings inoculated with 50 nematodes, population growth of the nematodes was suppressed and more time was needed for seedlings to die when pretreated with pH 3 SAR. This suggests that exposure to pH 3 SAR increased not only the progress of mortality, but also simultaneously enhanced the tolerance limit of the seedlings to the pinewood nematode – the critical value of physiological burden (represented as a product of time and initial nematode population) necessary to kill a seedling. Exposure to pH 2 SAR accelerated nematode reproduction in seedlings and increased seedling mortality irrespective of the number of nematodes inoculated.     

Acute hepatitis in a piglet experimentally inoculated with tissue homogenates from pigs with postweaning multisystemic wasting syndrome

Five 2 day-old colostrum-deprived piglets were inoculated with tissue homogenates from pigs with postweaning multisystemic wasting syndrome. One of the five piglets developed icterus and died 23 days post-inoculation. Histologic examination revealed acute hepatitis. Porcine circovirus type 2 (PCV-2) antigen and nucleic acid were detected in hepatocytes and phagocytic cells. Ultrastructurally, hepatocytes and phagocytic cells had large numbers of cytoplasmic inclusions, which were composed of electron-dense paracrystalline arrays of small non-enveloped viral particles approximately 17 nm in diameter. Apoptotic hepatocytes were confirmed by the TUNEL method and electron microscopic examination. These findings may indicate that hepatocellular necrosis is associated with replication of PCV-2. Apoptosis of hepatocytes also contributes to the pathogenesis of hepatic lesions in this case.     

Retardation of pine wilt disease symptom development in Japanese black pine seedlings exposed to simulated acid rain and inoculated withBursaphelenchus xylophilus

Three- or 4-year-old Japanese black pine seedlings were exposed to simulated acid rain (SAR) at pH 3 for two months, then inoculated with a virulent isolate (S 10) of pinewood nematodes. The experiments were repeated three times in 1996–1998. The exposure to SAR killed no seedlings, and retarded the development of disease symptoms in the seedlings inoculated with nematodes. In the experiments in 1996 and 1997, however, cessation of resin exudation, and primary decrease in the xylem conductivity occurred earlier in the seedlings exposed to SAR than in those exposed to tap water as a control. These results imply that acid rain at pH 3 influenced an increase in some resistance of Japanese black pine seedlings to pinewood nematode, and that this improved resistance could potentially overcome damage caused by acid rain. The exposure to SAR did not have any significant effect on the water relations of the seedlings itself, suggesting that retardation of the symptom development after the exposure to SAR could not be attributed to the improvement in the water relations. This work was supported in part by a Grants-in-Aid for Scientific Research from the Japan Forest Technical Association (JAFTA).     

Predicting chronosequential changes in carbon stocks of pachymorph bamboo communities in slash-and-burn agricultural fallow,northern Lao People’s Democratic Republic

In northern Lao People’s Democratic Republic, rising human population has drastically reduced the fallow period of slash-and-burn agriculture which has led to a considerable decrease in the carbon stock in these communities. We estimated chronosequential changes in the communities' carbon stocks, and established the relationship between the fallow period and fallow-period-average carbon stocks in three carbon pools of bamboo-dominated communities in hilly areas of the Luang Prabang Province, northern Lao People’s Democratic Republic. Based on measurements by destructive sampling, we devised a model and root-to-shoot ratios for estimating bamboo biomass. In six secondary plant communities established after slash-and-burn cropping, we estimated community biomass using the above model and others, and measured deadwood and litter stocks. The communities’ biomass and deadwood significantly increased with time after the last cropping and the former reached about 100 Mg ha⁻¹ after 15 years, whereas litter stocks did not show significant trends over time. Extending the fallow period from 2 to 5 years would increase fallow-period-average carbon stock from 14.2 to 25.1 Mg C ha⁻¹. The overstory height was significantly correlated with biomass, deadwood, and litter carbon stocks of these communities. Based on our findings, changes in a community’s carbon stocks can be estimated using the changes in overstory height, which should be taken into account in future studies to reduce uncertainty in estimating carbon stocks in tropical ecosystems.     

The effects of long‐term exposure to simulated acid rain on the development of pine wilt disease caused by Bursaphelenchus xylophilus

To elucidate the synergetic effects of acid rain on the development of pine wilt disease, we measured the sap flow rate in the stems and the chlorophyll content in the needles of 10‐year‐old Japanese black pine trees, Pinus thunbergii and 12‐year‐old Japanese red pine trees, Pinus densiflora, after exposure to simulated acid rain (SAR, pH 3) or tap water (TW, pH 6.3) as a control. The heat pulse method was used for the estimation of the sap flow rate. No apparent difference was found in the sap flow rate between the trees exposed to SAR and TW, but the chlorophyll content of needles at the end of the treatment was significantly higher in the trees exposed to SAR than in those exposed to TW. When the pinewood nematode, Bursaphelenchus xylophilus, the causal agent of pine wilt disease, was inoculated onto the Japanese black pines that had been exposed to SAR repeatedly for 1 year, the period to death was shortened. Japanese red pines that had been exposed to SAR for 2 years, however, did not show any development of symptoms after the nematode inoculation, suggesting that acid rain only affects pine wilt disease slightly, if at all.     

Antifungal limonoids from the fruits of Khaya senegalensis

Investigation of the fruits of Khaya senegalensis resulted in the isolation of three new mexicanolide limonoids containing a rare conjugated diene lactone system named seneganolide A (1), 2-hydroxyseneganolide A (2) and 2-acetoxyseneganolide A (3). Two known limonoids, 3-deacetyl-7-deacetoxy-7-oxokhivorin (4) and methyl 6-hydroxyangolensate (5), were also found. The structures of the new compounds were elucidated on the basis of spectral methods. The antifungal activity of compounds 1, 3 and 5 was tested against the fungus Botrytis cinerea.     

Condensation reactions of phenolic resins VII: catalytic effect of sodium bicarbonate for the condensation of hydroxymethylphenols

This article describes the catalytic effect of NaHCO₃ on condensation reactions of monomeric hydroxymethylphenols (HMPs) to elucidate the cure-acceleration mechanism. By comparison of the kinetics of self-condensations of HMPs, NaHCO₃ was proved to increase the reactivity of para-hydroxymethyl groups. The changes of ¹³C nuclear magnetic resonance (NMR) chemical shifts on each HMP system with the additive indicated that the addition of NaHCO₃ enhanced some molecular interactions between HMPs and NaHCO₃, facilitating a resonance effect that might play a similar role in dissociation of the phenolic hydroxyl groups of HMPs. In addition, computational modeling by molecular orbital calculations elucidated that hydrogen carbonate anion (HCO ₃ ⁻ ) forms an interaction between either the para-hydroxymethyl group and the phenolic hydroxyl group or between the two para-hydroxymethyl groups of HMPs by hydrogen bonds. From the experimental results, the authors proposed the mechanism of the catalytic action of NaHCO₃: it appears to be due to the delocalization of an electron initiated by the interaction of the para-hydroxymethyl groups and the phenolic hydroxyl of HMPs with HCO ₃ ⁻ through hydrogen bonds, which results in facilitating the formation of active species.     

Flonicamid, a novel insecticide with a rapid inhibitory effect on aphid feeding

Flonicamid (IKI220; N-cyanomethyl-4-trifluoromethylnicotinamide), a pyridinecarboxamide compound, is a novel systemic insecticide with selective activity against hemipterous pests, such as aphids and whiteflies, and thysanopterous pests. The purpose of this study is to clarify the biological properties of flonicamid against aphids. Flonicamid is very active against aphids, regardless of differences in species, stages and morphs. This compound inhibited the feeding behaviour of aphids within 0.5 h of treatment without noticeable poisoning symptoms such as convulsion, and this antifeeding activity was not recoverable until death. The nymphs born from adults exposed to flonicamid for 3 h showed high mortality. The effect of flonicamid on the feeding activity of an individual aphid was studied using electronic monitoring of insect feeding behaviour (EMIF). Although the treated aphid attached the head of its proboscis to the leaf surface, salivation and sap feeding were strongly inhibited. These results suggest that the main insecticidal mechanism of flonicamid is starvation based on the inhibition of stylet penetration to plant tissues.