This study aimed to prepare kombucha, a fermented tea beverage, containing Dendropanax morbiferus (DM) leaves and roots, and analyze its antioxidant and intracellular activities. We compared the pH change, total acidity, radical scavenging activity, and oxygen radical absorbance capacity (ORAC) of kombucha fermented with black tea alone and that with added DM leaves or roots during fermentation. Using RAW 264.7, we evaluated the effects of kombucha containing different DM parts on nitric oxide (NO) production and inflammation-related cytokine content in cells. Kombucha containing ethanol extracts of DM leaves (BTK-E-DML) and roots (BTK-E-DMR) showed higher radical scavenging activity and ORAC 3 d after fermentation than that prepared from black tea alone (BTK-Ori). In an in vitro experiment using RAW 264.7, samples were treated with 8 mg/mL kombucha considering cytotoxicity; the lipopolysaccharide (LPS)-induced NO content significantly reduced after BTK-E-DML and BTK-EDMR treatments compared with that after BTK-Ori treatment. Additionally, the levels of interleukin-6 and tumor necrosis factor-alpha, which were LPS-stimulated inflammatory cytokines, significantly decreased in cells treated with BTK-E-DML and BTK-E-DMR 15 d after fermentation compared with those treated with BTK-Ori. In conclusion, these results demonstrate that kombucha fermented with the leaves and roots of DM increases antioxidant activity and can significantly regulate inflammatory responses at the cellular level.

Six plant essential oils, vanillin, and their mixtures were tested for repellent activities and olfactory responses in a dengue virus vector mosquito, Aedes aegypti. Among the plant essential oils, cassia oil showed complete protection time (CPT) of 75 minutes. CPTs of lemongrass, lemoneucalyptus, xanthoxylum oils, and vanillin were within 30 minutes at 5% (0.21 mg/cm2) tested level, although their CPTs were not comparable to same concentration of DEET that showed 127.5 minutes of CPT. However, their repellency effects were significantly improved in two conditions; improving concentration up to 15% level (0.63 mg/cm2) and addition of vanillin. In bioassays using binary or tertiary mixtures with one or two essential oils with or without vanillin, the composition of 1:3:1 (v/v/w) consisted of lemongrass oil, xanthoxylum oil and vanillin provided 270 min-CPT. As a practical application, the mixture of 1:1:1 formulation containing lemongrass oil, xanthoxylum oil, and vanillin (v/v/w) was enclosed into the Viscopearl, porous cellulose beads that provide gradual release of volatile compounds. Efficiently, more than 90% of repellency for 2 hours was observed in cage and semi-field chamber tests using the formulation. In addition to behavioral assays, we subsequently examined how mosquitoes sense the blends of oils with vanillin by using electroantennogram (EAG) recording. Binary mixture with one oil and vanillin, which extended CPTs, showed no significant patterns of EAG alternation, while tertiary mixtures of oils and vanillin decreased patterns of EAG responses as an increase of vanillin contents in the mixture, implying further potential roles of vanillin as a synergist in mosquito repellency. Based on behavioral and electrophysiological data, cassia, rosemary, lemongrass, xanthoxylum, and lemoneucalyptus oils could provide the high possibility for development of commercial products for useful management strategies to control mosquitoes.

Olfaction as an important sensory modality in insects is essential for identification of hosts, mates, oviposition sites, and food resources in nature. In the cockroach, both olfactory sensitivity in the antennae and the formation of shortand long-term olfactory memories exhibit daily fluctuations that are regulated by the circadian system. An important problem is to characterize the signalling systems and molecules that are involved in this regulation of olfactory reception and olfactory behaviour. Recent results suggest that insect olfactory systems are modulated by both biogenic amines and neuropeptides. However, it remains elusive how these molecules modulate olfactory system in the peripheral systems. In the present study, our aim was to characterize the structure and organization of these signalling systems in the peripheral olfactory system of the American cockroach, Periplaneta americana. This work illuminated that tachykinin and its receptors regulate olfactory sensitivity in the antennae of the cockroach. Injections of tachykinin peptides caused decreases in the amplitude of the electroantennoogram (EAG), cells that produce tachykinin were localized in the antennae, and olfactory receptor neurons expressed tachykinin receptors. Interestingly, the tachykinin expressing cells also express receptors for the biogenic amine, octopamine and injections of octopamine also cause reductions in EAG amplitude. These results suggest that both octopaminergic and tachykinin peptide signalling pathways are important regulators of olfactory reception in the cockroach. We propose the hypothesis that octopamine regulates the release of tachykinin from cells in the antennae that, in turn, modulate the sensitivity of olfactory receptor neurons.

Aedes aegypti is a primary vector that transmits dengue and yellow fever around the world. To prevent the spreading and elimination of mosquitoes, insecticides and repellents like DEET (N, N-diethyl-m-methyl benzamide) have been broadly used. Even though DEET is considered as highly effective and proven protection against mosquito, it causes skin irritants and rashes, melts some synthetic plastics, and unpleasant smells. Therefore, there is a trend finding alternative mosquito repellents instead of using DEET. We tested repellent effects with plant essential oils and synergistic effects of those plant essential oils with additional vanillin, comparing them to DEET itself. Some of prepared mixtures showed better repellency than DEET. In addition, we evaluated the differences in the peripheral olfactory responses of Ae.aegypti females using EAG tests (electroantennogram). The aim of this test is to determined how the vanillin within plant essential oils or DEET acts in mosquito’s olfactory organs in aspect of molecular mechanisms. Revealing the novel function and localization of these putative repellent receptors may provide new insight into development of repellent as well as behavioral control agents in the future and contribute to understand the mechanism of processing patterns of repellent receptors in mosquitoes.

An olfactory system is one of the complicatedly-equipped sensory facilities in the insect sensory systems, which is most essential for insect olfactory-driven behaviors relevant to survival such as finding hosts, mates, oviposition sites, and food resources. These behaviors are mostly controlled by circadian rhythm. The american cockroach, Periplaneta americana, has been an ideal model to extensively study olfactory system associated with complex behavioral repertoires and circadian controls of certain behaviors, respectively. Even though it is known that olfactory-related physiology in peripheral and central olfactory systems seems to be highly variable by circadian rhythms, little is known about how these are controlled at the neuronal and molecular levels. It has been reported that the plasticity in the olfactory system is modulated by a set of neuropeptides. However, it remains still elusive how these neuropeptides and neuroendocrine system interact in the peripheral systems to change olfactory responses in cockroaches. Here, current study focuses on the localization of neuropeptides and their receptors by using in situ hybridization and immunostaining methods. Also, expression level of these genes are evaulated by qRT-PCR methods. Circadian fluctuation of these genes seem to be important neurotransmission machineries in the periphery. Our current study suggests that microcircuits of neuronal systems in the peripheral olfactory organ play an important in olfactory modulation by circadian rhythm