Hiroto Ohta

Development of an efficient process for the treatment of residual sludge discharged from an anaerobic digester in a sewage treatment plant

In order to reduce the discharge of residual sludge from an anaerobic digester, pre-treatment methods including low-pressure wet-oxidation, Fenton oxidation, alkali treatment, ozone oxidation, mechanical destruction and enzymatic treatment were evaluated and compared. VSS removal efficiencies of greater than 50% were achieved in cases of low-pressure wet-oxidation, Fenton oxidation and alkali treatment. Residual sludge from an anaerobic digester was pre-treated and subjected to thermophilic anaerobic digestion. As a result, the process of low-pressure wet-oxidation followed by anaerobic digestion achieved the highest VSS removal efficiency of 83%. The total efficiency of VSS removal of sewage sludge consisting of primary and surplus sludge would be approximately 92%, assuming that the VSS removal efficiency of sewage sludge is 50% in the anaerobic digester of the sewage treatment plant.

Isolation and Evaluation of PAH Degrading Bacteria

Polycyclic aromatic hydrocarbons (PAHs) degrading bacteria can be isolated from polluted coastal environments. They were close to Burkholderia fungorum and Mycobacterium gilvum, and had nidA, nidA3, pdoA2, and pcaH genes. In the pyrene and fluoranthene degradation experiment using a minimum medium, B. fungorum isolate no. 1 and M. gilvum isolate no. 13 were able to degrade 98.6% + 1.9 of pyrene after 20 days, and 99.9% ± 0.1 after a 16-day incubation period, respectively. Moreover, fluoranthene could be consumed by B. fungorum isolate no. 1 and M. gilvum isolate no. 13 at the level of 99.6% ± 0.7 after 24 days, and 100% after a 28-day incubation period, respectively.

Amitraz and its metabolite differentially activate α- and β-adrenergic-like octopamine receptors.

BACKGROUND Amitraz is a formamidine acaricide and insecticide used to control ticks, mites and fleas. N2 -(2,4-Dimethylphenyl)-N1 -methyformamidine (DPMF), a metabolite of amitraz, is thought to be an active agent that exerts acaricidal and insecticidal effects by acting as an agonist on octopamine receptors. The emergence of cattle ticks resistant to amitraz is a serious problem that requires urgent attention. The objective of this research was to determine which type of octopamine receptor is the primary target of amitraz and thereby understand the molecular mechanisms of action and resistance to amitraz. RESULTS Amitraz and DPMF potently activated Bombyx mori α- and β-adrenergic-like octopamine receptors (α- and β-AL OARs) that were stably expressed in HEK-293 cells. Notably, DPMF elevated intracellular cAMP levels, with an EC50 of 79.6 pm in β-AL OARs, the transcripts of which were prevalently and widely localised in B. mori body parts. Furthermore, DPMF elevated the intracellular Ca2+ levels, with an EC50 of 1.17 nm in α-AL OARs. CONCLUSION Although both amitraz and DPMF acted as OAR agonists, the metabolite DPMF was more potent than amitraz and differentially activated α- and β-AL OARs. The present findings provide a basis for studies to examine the mechanism of amitraz resistance and to develop novel acaricides and insecticides.

Tyramine and β-phenylethylamine, from fermented food products, as agonists for the human trace amine-associated receptor 1 (hTAAR1) in the stomach

The aromatic amines tyramine and β-phenylethylamine are abundant in fermented foods. Recently, a family of human trace amine-associated receptors (hTAARs) was discovered that responds to these compounds. This study examined the expression of hTAAR genes in five human organs. Among them, the stomach expressed hTAAR1 and hTAAR9. Interestingly, more hTAAR1 was expressed in the pylorus than in the other stomach regions. The CRE-SEAP reporter assay revealed that only hTAAR1 functioned as a Gs-coupled receptor in response to tyramine and β-phenylethylamine stimulation. The β-phenylethylamine-mediated hTAAR1 activity could be potentiated using 3-isobutyl-1-methylxanthine. These data suggest that tyramine and β-phenylethylamine in fermented foods act at hTAAR1 as agonists in the pylorus of stomach. Graphical abstract hTAAR1 expression in stomach regions and a possible physiological action of tyramine and β-phenylethylamine from fermented foods on hTAAR1 expressed in the pylorus.

A secreted placental alkaline phosphatase-based reporter assay system for screening of compounds acting at an octopamine receptor stably expressed in a mammalian cell line

Octopamine receptors are attractive insecticide targets. To screen compounds acting at octopamine receptors simply and rapidly, we constructed a chemiluminescent reporter gene assay system that detects secreted placental alkaline phosphatase transcriptionally regulated by the cAMP response element for a silkworm octopamine receptor. This system proved useful in high-throughput screening to develop octopamine receptor-specific insecticides.

Elevenin regulates the body color through a G protein-coupled receptor NlA42 in the brown planthopper Nilaparvata lugens

The neuropeptide elevenin and similar neuropeptide precursors are common in some invertebrates but their physiological function in most species has not been explored. The brown planthopper, Nilaparvata lugens (Stål) has an elevenin-like peptide and a G protein-coupled receptor (GPCR) NlA42 that is homologous to the elevenin receptor of the annelid Platynereis dumerilii. RNA interference (RNAi)-mediated knockdown of either Nl-elevenin or the NlA42 gene resulted in cuticle melanization. Ion transport peptide (ITP) also induces melanization, but unlike ITP, knockdown of NlElevenin and NlA42 did not have any effect on wing expansion or activity after eclosion. In wild condition macropterous individuals show a darker body color when compared with brachypterous individuals, but RNAi experiments suggest that insulin-signaling and Nl-elevenin signaling regulate wing morph and body color independently. NlElevenin was predominantly expressed in the brain while NlA42 was highly expressed in the abdominal integument and brain. A signal Calcium assays using aequorin indicated that NlA42 heterologously expressed in HEK293 cells exhibited responses to synthetic Nl-elevenin peptide from concentrations as low as 10-9M. These results suggest that neuropeptide Nl-elevenin is involved in the regulation of melanization through its receptor NlA42. This is the first report of a physiological function for elevenin-like peptides in insects.