Rhynchophorus ferrugineus midgut cell line to evaluate insecticidal potency of different plant essential oils.

Cell cultures can be a potent and strong tool to evaluate the insecticidal efficiency of natural products. Plant essential oils have long been used as the fragrance or curative products around the world which means that they are safer to be used in close proximity of humans and mammals. In this study, a midgut cell line, developed from Rhynchophorus ferrugineus (RPW-1), was used for screening essential oils from nine different plants. Assays revealed that higher cell mortality was observed at 500 ppm which reached to 86, 65, 60, 59, 56, 54, 54, 53, and 53%, whereas lowest cell mortality at 1 ppm remained at 41, 23, 20, 17, 16, 15, 14, 13, and 10%, for Azadirachta indica, Piper nigrum, Mentha spicata, Cammiphora myrrha, Elettaria cardamomum, Zingiber officinale, Curcuma longa, Schinus molle, and Rosmarinus officinalis, respectively. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell proliferation assay revealed the percentage of cell growth inhibition was highest at 500 ppm and remained at 48, 45, 42, 37, 34, 29, 24, 22, and 18% against A. indica, P. nigrum, M. spicata, C. myrrha, E. cardamomum, Z. officinale, C. longa, S. molle, and R. officinalis, respectively. Lowest LC50 value (7.98 ppm) was found for A. indica, whereas the highest LC50 (483.11 ppm) was against R. officinalis. Thus, in this study, essential oils of A. indica exhibited the highest levels of toxicity, whereas those from R. officinalis exhibited the lowest levels of toxicity toward RPW-1 cells.

Apolipophorin III and transmission electron microscopy as toxicity indicators for harmaline and tea saponin in Spodoptera exigua (Noctuidae: Lepidoptera).

Apolipophorin III, traditionally known for lipid transport in insects is fairly established as toxicity indicator against harmaline and tea saponin during this study. Apolipophorin III expressed in the hemolymph and midgut tissues of 3rd, 4th, 5th larval instars and pupae of Spodoptera exigua. Apolipophorin III presence was further confirmed by achieving its partial cDNA (Genbank accession no. FJ606822) of 448bp. qRT PCR revealed that tea saponin resulted in significant reduction of gene expression in 3rd and 4th larval instars but increased in 5th instar as compared to control. Harmaline caused gradual increase of gene expression in 3rd, 4th and 5th instars after feeding on the treated diet. Fifth instar larvae synonymously resulted in the highest gene expressions against both the biochemicals. After the injection of harmaline and tea saponin abrupt increase in gene expression of 4th, 5th larval instar and pupae was observed as compared to control treatment. Transmission electron microscopy of midgut epithelium after being fed with harmaline and tea saponin depicted certain cytological changes. Harmaline treatment lead to cytoplasm vacuolization, mitochondrial disruption, spherocrystals with concentric layers, irregular nucleus and floating nuclei in cytoplasm. Tea saponin treatment resulted in denser cytoplasm, higher intracellular osmotic concentration and reduced complement of apical microvilli. Cells were found to have only a few mitochondria and glycogen deposits in comparison to control treatment.

Biodegradation of beta-cypermethrin and 3-phenoxybenzoic acid by a novel Ochrobactrum lupini DG-S-01.

A newly isolated bacterium DG-S-01 from activated sludge utilized beta-cypermethrin (beta-CP) and its major metabolite 3-phenoxybenzoic acid (3-PBA) as sole carbon and energy source for growth in mineral salt medium (MSM). Based on the morphology, physio-biochemical characteristics, and 16S rDNA sequence analysis, DG-S-01 was identified as Ochrobactrum lupini. DG-S-01 effectively degraded beta-CP with total inocula biomass A(590 nm) = 0.1-0.8, at 20-40 °C, pH 5-9, initial beta-CP 50-400 mg L(-1) and metabolized to yield 3-PBA leading to complete degradation. Andrews equation was used to describe the special degradation rate at different initial concentrations. Degradation rate parameters q(max), K(s) and K(i) were determined to be 1.14 d(-1), 52.06 mg L(-1) and 142.80 mg L(-1), respectively. Maximum degradation was observed at 30 °C and pH 7.0. Degradation of beta-CP was accelerated when MSM was supplemented with glucose, beef extract and yeast extract. Studies on biodegradation in liquid medium showed that over 90% of the initial dose of beta-CP (50 mg L(-1)) was degraded under the optimal conditions within 5d. Moreover, the strain also degraded beta-cyfluthrin, fenpropathrin, cyhalothrin and deltamethrin. These results reveal that DG-S-01 may possess potential to be used in bioremediation of pyrethroid-contaminated environment.