Tomomitsu Satho

Indoor-breeding of Aedes albopictus in Northern Peninsular Malaysia and its potential epidemiological implications.

Background The mosquito Ae. albopictus is usually adapted to the peri-domestic environment and typically breeds outdoors. However, we observed its larvae in most containers within homes in northern peninsular Malaysia. To anticipate the epidemiological implications of this indoor-breeding, we assessed some fitness traits affecting vectorial capacity during colonization process. Specifically, we examined whether Ae. albopictus exhibits increased survival, gonotrophic activity and fecundity due to the potential increase in blood feeding opportunities. Methodology/Principal Findings In a series of experiments involving outdoors and indoors breeding populations, we found that Ae. albopictus lives longer in the indoor environment. We also observed increased nighttime biting activity and lifetime fecundity in indoor/domestic adapted females, although they were similar to recently colonized females in body size. Conclusion/Significance Taken together these data suggest that accommodation of Ae. albopictus to indoor/domestic environment may increase its lifespan, blood feeding success, nuisance and thus vectorial capacity (both in terms of increased vector-host contacts and vector population density). These changes in the breeding behavior of Ae. albopictus, a potential vector of several human pathogens including dengue viruses, require special attention.

Purification and characterization of intracellular α-l-rhamnosidase from Pseudomonas paucimobilis FP2001

Abstractα-l-Rhamnosidase was extracted and purified from the cells of Pseudomonas paucimobilis FP2001 with a 19.5% yield. The purified enzyme, which was homogeneous as shown by SDS-PAGE and isoelectric focusing, had a molecular weight of 112,000 and an isoelectric point of 7.1. The enzyme activity was accelerated by Ca2+ and remained stable for several months when stored at –20 °C. The optimum pH was 7.8; the optimum temperature was 45 °C. The Km, Vmax and kcat for p-nitrophenyl α-l-rhamnopyranoside were 1.18 mM, 92.4 μM · min–1 and 117,000 · min–1, respectively. Examination of the substrate specificity using various synthetic and natural l-rhamnosyl glycosides showed that this enzyme had a relatively broader substrate specificity than those reported so far.

Identification and molecular characterization of a new nonsegmented double-stranded RNA virus isolated from Culex mosquitoes in Japan.

Two infectious agents were isolated from Culex species mosquitoes in Japan and were identified as distinct strains of a new RNA virus by a method for sequence-independent amplification of viral nucleic acids. The virus designated Omono River virus (OMRV) replicated in mosquito cells in which it produced a severe cytopathic effect. Icosahedral virus particles of approximately 40 nm in diameter were detected in the cytoplasm of infected cells. The OMRV genome was observed to consist of a nonsegmented, 7.6-kb double-stranded RNA (dsRNA) and contain two overlapping open reading frames (ORFs), namely ORF1 and ORF2. ORF1 was found to encode a putative dsRNA-binding protein, a major capsid protein, and other putative proteins, which might be generated by co- and/or post-translational processing of the ORF1 polyprotein precursor, and ORF2 was found to encode a putative RNA-dependent RNA polymerase (RdRp), which could be translated as a fusion with the ORF1 product by a -1 ribosomal frameshift. Phylogenetic analysis based on RdRp revealed that OMRV is closely related to penaeid shrimp infectious myonecrosis virus and Drosophila totivirus, which are tentatively assigned to the family Totiviridae. These results indicated that OMRV is a new member of the family of nonsegmented dsRNA viruses infecting arthropod hosts, but not fungal or protozoan hosts.

The effects of simulated rainfall on immature population dynamics of Aedes albopictus and female oviposition

Larvae of Aedes albopictus Skuse typically inhabit natural and artificial containers. Since these larval habitats are replenished by rainfall, Ae. albopictus may experience increased loss of immature stages in areas with high levels of rainfall. In this study, we investigated the effects of rainfall and container water level on population density, and oviposition activity of Ae. albopictus. In field and laboratory experiments, we found that rainfall resulted in the flushing of breeding habitats. Excess rain negatively impacted larval and pupal retention, especially in small habitats. When filled with water to overflowing, container habitats were significantly repellent to ovipositing females. Taken together, these data suggest that rainfall triggers population loss of Ae. albopictus and related species through a direct detrimental effect (flushing out) and an indirect effect (ovipositional repellency).

Insecticide susceptibility of the dengue vector Aedes aegypti (Diptera: culicidae) in Makkah City, Saudi Arabia

Objective To examine the insecticide susceptibility of Aedes aegypti (Ae. aegypti) from various sites in Makkah City, Saudi Arabia. Methods This was examined based on WHO standard procedures. Results The larvae of Ae. aegypti were susceptible to all larvicides examined, but this susceptibility was more pronounced in wild populations, which tended to show tolerance to icon. Icon was the most effective larvicide with LC50 values of 0.007 ppm and 0.012 ppm for the laboratory and field strains, respectively. Ae. aegypti adults exposed to lambda-cyhalothrin showed a low mortality rate in comparison with those exposed to deltamethrin and cyfluthrin. Conclusions The results of the present study indicate differential susceptibility between field and laboratory larval populations. Wild larvae are less susceptible to insecticide treatments than their laboratory-bred counterparts. Taken together, these results suggest that tolerance and the tendency toward resistance to commonly used insecticides are present in Ae. aegypti populations throughout Makkah City, Saudi Arabia.

Changing Domesticity of Aedes aegypti in Northern Peninsular Malaysia: Reproductive Consequences and Potential Epidemiological Implications

Background The domestic dengue vector Aedes aegypti mosquitoes breed in indoor containers. However, in northern peninsular Malaysia, they show equal preference for breeding in both indoor and outdoor habitats. To evaluate the epidemiological implications of this peridomestic adaptation, we examined whether Ae. aegypti exhibits decreased survival, gonotrophic activity, and fecundity due to lack of host availability and the changing breeding behavior. Methodology/Principal Findings This yearlong field surveillance identified Ae. aegypti breeding in outdoor containers on an enormous scale. Through a sequence of experiments incorporating outdoors and indoors adapting as well as adapted populations, we observed that indoors provided better environment for the survival of Ae. aegypti and the observed death patterns could be explained on the basis of a difference in body size. The duration of gonotrophic period was much shorter in large-bodied females. Fecundity tended to be greater in indoor acclimated females. We also found increased tendency to multiple feeding in outdoors adapted females, which were smaller in size compared to their outdoors breeding counterparts. Conclusion/Significance The data presented here suggest that acclimatization of Ae. aegypti to the outdoor environment may not decrease its lifespan or gonotrophic activity but rather increase breeding opportunities (increased number of discarded containers outdoors), the rate of larval development, but small body sizes at emergence. Size is likely to be correlated with disease transmission. In general, small size in Aedes females will favor increased blood-feeding frequency resulting in higher population sizes and disease occurrence.

Trace metal zinc stimulates secretion of antimicrobial peptide LL-37 from Caco-2 cells through ERK and p38 MAP kinase.

Infectious diseases, especially, diarrhoea, are responsible for high mortality rates in developing countries. Zinc supplementation shows beneficial effects against such diseases, but the mechanism of action is poorly understood. Here, we examined whether zinc supplementation can improve mucosal innate immunity through induction of antimicrobial peptide secretion from intestinal epithelial cells. Zinc was found to induce secretion of the antimicrobial peptide LL-37 from Caco-2 cell in a dose (0.63±0.09ng/mL and 0.54±0.06ng/mL at 20μM and 50μM respectively) and time dependent manner. LL-37 secretion increased immediately (1h) after exposure to 20μM Zn (0.29±0.04ng/mL), which continued up to 48h of exposure (0.58±0.05ng/mL). Zinc induces the phosphorylation of ERK and p38 MAP kinase and regulates LL-37 secretion through these MAP kinases. Zinc supplementation may have beneficial effects on mucosal innate immunity via secretion of LL-37.

Cloning, Sequence Analysis, and Expression of the Gene Encoding Sphingomonas paucimobilis FP2001 α- L -Rhamnosidase

The gene (rhaM) encoding the α-L-rhamnosidase of Sphingomonas paucimobilis FP2001 was cloned, sequenced, and expressed in Escherichia coli. The rhaM consisted of 3,354 nucleotides and had a promoter and Shine-Dalgarno sequences typical in bacteria. The rhaM encoding a protein (Rham) deducted from the sequence consisted of 1,117 amino acids and had a putative signal peptide of 25 amino acids. Rham has no similarity to other known rhamnosidases. Rham has a sugar-binding domain of glycoside hydrolase family 2, which has been well conserved in β-glucuronidase, β-mannosidase, and β-galactosidase, in its C-terminal region. Rham is possibly a member of a new bacterial subfamily in glycoside hydrolase family 78 (α-L-rhamnosidase). RT-PCR analysis of rhaM mRNA indicated that the induction of α-L-rhamnosidase by the addition of L-rhamnose occurred on the transcriptional level.

Colonized Aedes albopictus and its sexual performance in the wild: implications for SIT technology and containment

BackgroundMating is a physiological process of crucial importance underlying the size and maintenance of mosquito populations. In sterile and incompatible insect technologies (SIT and IIT), mating is essential for mass production, persistence, and success of released individuals, and is a central parameter for judging the effectiveness of SIT/IIT programs. Some mosquitoes have an enormous reproductive potential for both themselves and pathogens and mating may contribute to persistence of infection in nature. As Aedes albopictus can transmit flaviviruses both sexually and horizontally, and as infected insects are usually derived from laboratory colonies, we investigated the implications of mating between a long-term laboratory colony of Ae. albopictus and wild populations.MethodsThrough a series of mating experiments, we examined the reproductive outcomes of sexual cross-affinity between laboratory-raised and wild adults of Ae. albopictus.ResultsThe results indicated appreciable mating compatibility between laboratory-reared and wild adults, and equivalent levels of egg production among reciprocal crosses. We also observed comparable larval eclosion in lab females mated with wild males, and increased adult longevity in female offspring from wild females|×|laboratory males crosses.ConclusionsTaken together, these data suggest that Ae. albopictus can preserve its reproductive fitness over a long period of time in the laboratory environment and has valuable attributes for SIT application. These observations together with the ability to successfully inseminate heterospecific females indicate the potential of Ae. albopictus to act as an ecological barrier if non-sterilized males are massively released in areas occupied by Aedes aegypti. The observed substantial reproductive fitness combined with the capability to reproduce both, itself and viruses illustrates the potential of Ae. albopictus to pose a serious threat if infected and released accidentally.

Characterization of Dak Nong virus, an insect nidovirus isolated from Culex mosquitoes in Vietnam.

In this study, we isolated and characterized an insect nidovirus from the mosquito Culex tritaeniorhynchus Giles (Diptera: Culicidae) in Vietnam, as an additional member of the new family Mesoniviridae in the order Nidovirales. The virus, designated “Dak Nong virus (DKNV),” shared many characteristics with Cavally virus and Nam Dinh virus, which have also been discovered recently in mosquitoes, and these viruses should be considered members of a single virus species, Alphamesonivirus 1. DKNV grew in cultured mosquito cells but could not replicate in the cultured vertebrate cells tested. N-terminal sequencing of the DKNV structural proteins revealed two posttranslational cleavage sites in the spike glycoprotein precursor. DKNV is assumed to be a new member of the species Alphamesonivirus 1, and the current study provides further understanding of viruses belonging to the new family Mesoniviridae.