Toxicity, antifeedant and physiological effects of trans-anethole against Hyphantria cunea Drury (Lep: Arctiidae).

Plant secondary metabolites are currently known to interfere with basic metabolic, behavioral and physiological processes of insects. In the current study, the biological and physiological effects of trans-anethole were investigated against Hyphantria cunea Drury. The bioassay data demonstrated the high toxicity of trans-anethole against the fourth-instar larvae with the LC30, LC50 and LC90 values of 0.72, 1.41 and 7.20 µL/mL, respectively. Also the concentrations of LC30 and LC50 showed 53 and 87% feeding deterrency against the larvae. The biochemical experiments revealed that oral exposure of trans-anethole decreased the activities of digestive enzymes, acetylcholinesterase and the contents of energy reserves while, it induced the activities of detoxifying and antioxidant enzymes compared to control. In fact, trans-anethole induced the inhibition of digestion and AChE activities accompanied by imbalance in metabolic and oxidative processes so it may be recommended as a potent biopesticide in control of H. cunea populations.

Immunological interactions of Chilo suppressalis Walker (Lepidoptera: Crambidae) with the native entomopathogenic fungi.

Entomopathogenic fungi can attack many insect hosts and have been applied as the eco-friendly alternatives to synthetic chemicals for the control of pests. Insects have developed different defense systems encountering entomopathogens including humoral and cellular immune responses. In the present study, injection of some native entomopathogenic fungi to the Chilo suppressalis Walker larvae resulted in an enhancement of the cellular and antimicrobial defenses. The numbers of total and differential hemocytes increased rapidly in the first 3 and 6 h but those gradually reduced 12 and 24 h post-injections. The nodule formation and phenoloxidase activity increased at the time intervals after fungal infection. A similar trend was found in the transcription of antimicrobial peptides including attacin1 and 2, cecropin1 and 2, gallerimycin, defensin, lysozyme, and prophenoloxidase-activating proteinase-3 during infection fungi. In all cases, the target gene transcription was upper in the larvae injected by the fungi than that of control larvae. These results may elucidate better knowledge on the interaction of the fungi present in agroecosystems with the target insect pest.

Screening and Virulence of the Entomopathogenic Fungi Associated with Chilo suppressalis Walker.

The current study aimed to explore the entomopathogenic fungi associated with the larvae of Chilo suppressalis Walker, a serious pest of rice, in northern Iran. The collected specimens were cultured and identified through morphological and molecular methods. The 38 specimens were identified by microscopic examination and genetic sequencing of the ITS region as follows: twenty-one isolates of Beauveria bassiana, five isolates of Akanthomyces lecanii, four isolates of Akanthomyces muscarious, three isolates of Metarhizium anisopliae, two isolates of Hirsutella subulata, two isolates of Trichoderma sp. and one isolate of Aspergillus sp. All the identified isolates were treated on the larvae through bioassay, evaluating the amount of hydrophobin and the activities of proteases, chitinases and lipase to find their virulence. Moreover, the percentage of thermotolerant and cold activity of the isolates were tested to determine their environmental persistence. The overall results revealed the isolates of B. bassiana, including BBRR1, BBAL1 and BBLN1 as the most virulent and environmental adaptive isolates among the fungi associated with C. suppressalis.

Toxicity and physiological effects of ajwain (Carum copticum, Apiaceae) essential oil and its major constituents against Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae).

The effects of Ajwain essential oil and its major constituents were investigated against newly fourth instar larvae of Tuta absoluta, a major pest of tomato cropping in Iran and other parts of the world. The study included individual efficacy of each compound and determination of their synergistic or antagonistic efficacy. We also determined the activities of detoxifying enzymes in treated insects. According to the results, the Ajwain essential oil and thymol showed the highest toxicity against T. absoluta larvae (LC50 = 6.14 and 7.72 µL/mL, respectively, and LC30 = 3.05 and 3.57 µL/mL, respectively). The ɤ-terpinene and p-cymene had lesser toxicity compared to other compounds. The larvicidal potency increased by binary mixtures of active constituents, such as thymol + p-cymene and thymol + É¤-terpinene. On the other hand, ɤ-terpinene + p-cymene mixture demonstrated antagonistic effect on the larvae. The esterases and glutathione S-transferases were significantly increased in all treatments. Finally, our results revealed a significant inhibition of AChE activity in the treated larvae by all treatments except for ɤ-terpinene + p-cymene. In vitro experiments representing AChE inhibition with IC50 values were recorded 0.370, 0.457, 0.528, 1.094 and 1.323 µL/mL for thymol + p-cymene, thymol + É¤-terpinene, thymol, ɤ-terpinene and p-cymene, respectively. Thymol demonstrated significant potential as a controlling agent of T. absoluta larvae, both larvicidal, compatible with other compounds with strong AChE inhibition properties. These finding could pave the way for development of new botanicals based on EO constituents which should be completed with preparation of an efficient formulation and field trials.

Effects of α-pinene, trans-anethole, and thymol as the essential oil constituents on antioxidant system and acetylcholine esterase of Ephestia kuehniella Zeller (Lepidoptera: Pyralidae).

The current study aimed to determine the potential effects of three essential oil constituents, α-pinene, trans-anethole, and thymol, on antioxidant system and acetylcholine esterase (AChE) of Ephestia kuehniella Zeller. The 4th instar larvae were initially fed on an artificial diet containing an LC50 concentration of each above-mentioned compounds separately prior to being undertaken for sample preparation and biochemical assays. The significant higher activities of superoxide dismutase, peroxidase and catalase were observed in the treated-larvae at both time intervals. Similar findings were found in the activity of glutathione S-transferase by using both reagents. Although activities of ascorbate peroxidase and glycerol-6-phosphate dehydrogenase increased in the treated larvae by all constituents while glycerol-6-phosphate dehydrogenase showed no statistically different activity among the larvae fed on α-pinene, trans-anethole, and thymol. The concentration of malondialdehyde and the ratio of oxidized (RSSR) to reduced (RSH) thiols showed statistical differences among control and treated larvae except for time interval of 24 h regarding the ratio of RSSR/RSH. Finally, our results demonstrated a significant decrease of AChE activity in the treated larvae by all constituents after 24 h while no statistical differences were found between control and trans-anethole after 24 h. Also, in vitro analysis revealed significant inhibition of AChE representing IC50 values of 0.864, 0.490 and 0.137 µl/ml for α-pinene, trans-anethole, and thymol, respectively. These results determined significant effects of administered constituents on induction of antioxidant system and inhibition of a nervous system component which expand our knowledge on physiological turbulences due to essential oil treatment.