Assessment of sublethal and transgenerational effects of spirotetramat, on population growth of cabbage aphid, Brevicoryne brassicae L. (Hemiptera: Aphididae).

The cabbage aphid (Brevicoryne brassicae L.) is a devastating pest of cruciferous crops causing economic damage worldwide and notably owing to its increasing resistance to commonly used pesticides. Such resistance prompts the development of integrated pest management (IPM) programs that include novel pesticides being effective against the aphids. Spirotetramat is a novel insecticide used against sap-sucking insect pests, particularly aphids. This study evaluated the toxicity of spirotetramat to adult apterous B. brassicae after 72 h using the leaf dipping method. According to the toxicity bioassay results, the LC50 value of spirotetramat to B. brassicae was 1.304 mgL-1. However, the sublethal concentrations (LC5 and LC15) and transgenerational effects of this novel insecticide on population growth parameters were estimated using the age-stage, two-sex life table theory method. The sublethal concentrations (LC5; 0.125 mgL-1 and LC15; 0.298 mgL-1) of spirotetramat reduced the adult longevity and fecundity of the parent generation (F0). These concentrations prolonged the preadult developmental duration while decreasing preadult survival, adult longevity and reproduction of the F1 generation. The adult pre-reproductive period was also extended by spirotetramat treatment groups. Subsequently, the population growth parameters such as the intrinsic rate of increase r, finite rate of increase λ and net reproductive rate R 0 of the F1 generation were decreased in spirotetramat treatment groups whereas, the mean generation time T of the F1 generation was not affected when compared to the control. These results indicated the negative effect of sublethal concentrations of spirotetramat on the performance of B. brassicae by reducing its nymphal survival, extending the duration of some immature stages and suppressing the population growth of B. brassicae. Overall, we demonstrated that spirotetramat is a pesticide showing both sublethal activities, and transgenerational effects on cabbage aphid; it may be useful for implementation in IPM programs against this aphid pest.

Establishment of Wolbachia infection in Aedes aegypti from Pakistan via embryonic microinjection and semi-field evaluation of general fitness of resultant mosquito population.

BACKGROUND: Dengue is a mosquito-borne viral disease that is mainly spread by Aedes aegypti. It is prevalent on five continents, predominantly in tropical and sub-tropical zones across the world. Wolbachia bacteria have been extensively used in vector control strategies worldwide. The focus of the current study was to obtain a natural population of Ae. aegypti harbouring Wolbachia and to determine the impact of this bacteria on the new host in a semi-field environment. METHODS: Wolbachia-infected Aedes albopictus was collected from the city of Lahore, Punjab, Pakistan, and Wolbachia were successfully introduced into laboratory-reared Ae. aegypti via embryonic microinjection. The stable vertical transmission of wAlbB in the host population was observed for eight generations, and the impact of Wolbachia on the general fitness of the host was evaluated in semi-field conditions. RESULTS: In the laboratory and semi-field experiments, wAlbB Wolbachia presented a strong cytoplasmic incompatibility (CI) effect, evidenced as zero egg hatching, in crosses between Wolbachia-infected males and wild (uninfected) females of Ae. aegypti. Wolbachia infection had no noticeable impact on the general fitness (P > 0.05), fecundity, body size (females and males) and mating competitiveness of the new host, Ae. aegypti. However, there was a significant decrease in female fertility (egg hatch) (P < 0.001). In addition, under starvation conditions, there was a remarkable decrease (P < 0.0001) in the life span of Wolbachia-infected females compared to uninfected females (4 vs. > 5 days, respectively). CONCLUSIONS: Wolbachia strain wAlbB has a great potential to control the dengue vector in Ae. aegypti populations by producing 100% CI with a limited burden on its host in natural field conditions. This strain can be used as a biological tool against vector-borne diseases.

Sublethal effects of bistrifluron on key biological traits, macronutrients contents and vitellogenin (SeVg) expression in Spodoptera exigua (Hübner).

The beet armyworm, Spodoptera exigua, is a highly polyphagous pest originated from Southeast Asia but has spread globally, attacking economically important crops and fruits. Bistrifluron insecticide is one of the highly active insect growth regulators that has been reported to inhibit development and longevity in other lepidopteran species and could be used in the control of S. exigua. In the present study, the age-stage, two-sex life table technique was applied to assess the sublethal effects of bistrifluron on biological traits and vitellogenin gene (SeVg) expression when 2nd instar larvae fed to sublethal concentrations (LC10, LC20 and LC40) of bistrifluron. Mean generation time from eggs to adults was longer at LC40 (37.79 ± 0.81 d) and LC20 (37.04 ± 0.72) compared to the LC10 (36.89 ± 0.63 d) and control groups (36.07 ± 0.38 d). Fecundity of female at LC40 (279.17 ± 42.8 eggs), LC20 (347 ± 35.4 eggs) and LC10 (411.58 ± 42.38 eggs) were significantly lower than the control treatment (532.47 ± 7.13). Furthermore, the lower intrinsic rates of increase (LC40; r = 0.1207 ± 0.009, LC20; r = 0.1329 ± 0.009 and LC10; r = 0.14398 ± 0.009 compared to the control r = 0.164 ± 0.0076), was observed along with significantly extended mean generation times (LC40; T = 34.825 ± 0.317 days, LC20; T = 33.27 ± 0.368 days and LC10; T = 31.899 ± 0.398 days compared to the control 30.927 ± 0.255 days). Furthermore, the contents of energy reserve macronutrients (carbohydrate, lipid and protein) significantly reduced in dose and time dependent manner in treated insects as compared to control. Furthermore, the expression level of SeVg mRNA significantly decreased by 43.8% in the female adults when one-day-old second instar larvae were treated with sublethal concentrations of bistrifluron in comparison with the control. Documenting these sublethal effects is a vital, and often overlooked factor, in assessing the overall efficacy of insecticides in the management of pest populations.

Author Correction: Impact of low lethal concentrations of buprofezin on biological traits and expression profile of chitin synthase 1 gene (CHS1) in melon aphid, Aphis gossypii.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Expression and functional analysis of P450 gene induced tolerance/resistance to lambda-cyhalothrin in quercetin fed larvae of beet armyworm Spodoptera exigua (Hübner).

Beet armyworm, Spodoptera exigua (Hübner) is an agronomical important and most devastating polyphagous pest that damages a variety of crops around the globe including China. Quercetin is one of the abundant dietary flavonoids and the important defense allelochemicals in plants. Therefore, the changes in insect detoxification enzymes activities in response to plants allelochemicals may result increased the sensitivity to insecticides. In this study, we examined the induced effect of quercetin on larval tolerance to lambda-cyhalothrin in S. exigua. Application of cytochrome P450 inhibitor piperonyl butoxide (PBO) significantly synergized the lambda-cyhalothrin toxicity in quercetin-fed S. exigua larvae. Moreover, larval weight significantly reduced in quercetin, lambda-cyhalothrin, and quercetin + lambda-cyhalothrin treatment. Furthermore, our results showed that the P450 detoxification enzyme effectively increased in all treatments as compared to the control. Quantitative Real-time PCR analysis revealed that expression level of CYP6AE10 significantly upregulated in larvae treated with quercetin, lambda-cyhalothrin and quercetin + lambda-cyhalothrin in the midgut and fat body respectively. In addition, RNAi mediated knockdown of CYP6AE10 in S. exigua larvae significantly decreased the transcription level of target cytochrome P450 gene followed by the exposure with quercetin, lambda-cyhalothrin, and quercetin + lambda-cyhalothrin. Similarly, the knockdown of CYP6AE10 by the injection of dsRNA led to increased mortality after the treatment with respective chemicals. Overall, these data showed that P450s might possibly play an important role in the metabolic adaptation of S. exigua larvae to its host plant defense allelochemicals as well as insecticides. In conclusion, S. exigua can take benefit from its host plant's secondary metabolites to elaborate its defense against synthetic insecticides.

RNA interference-mediated knockdown of a cytochrome P450 gene enhanced the toxicity of α-cypermethrin in xanthotoxin-fed larvae of Spodoptera exigua (Hübner).

The beet armyworm (Spodoptera exigua) is a highly polyphagous agricultural pest that is distributed worldwide. However, the adaptive mechanisms of S. exigua for various insecticides and defensive substances in host plants are unknown. Insect P450 monooxygenases play an important role in the detoxification of plant toxins and insecticides, leading to insecticides resistance. We investigated the induced effects of xanthotoxin exposure on detoxification enzyme activity and larval tolerance to α-cypermethrin in S. exigua. Our results showed that the lethal concentration (LC50) of α-cypermethrin for xanthotoxin-exposed larvae was 2.1-fold higher than in the control. Moreover, cytochrome P450 enzyme activity was significantly elevated by upregulation of P450 genes in treated larvae. RT-qPCR results showed that CYP9A10 expression level was significantly increased in all treatments, while maximal expression level was observed in xanthotoxin+α-cypermethrin-fed larvae. RNAi-mediated silencing of CYP9A10 further increased mortality by 18%, 26% and 35% at 48 h and by 27%, 43% and 55% at 72 h when larvae were exposed to diets containing chemicals as compared to the control. The results show that CYP9A10 might play an important role in xanthotoxin and α-cypermethrin detoxification in S. exigua. RNAi-mediated silencing could provide an effective synergistic agent for pest control or insecticide resistance management.

Impact of low lethal concentrations of buprofezin on biological traits and expression profile of chitin synthase 1 gene (CHS1) in melon aphid, Aphis gossypii.

Buprofezin, a chitin synthesis inhibitor that can be used for the control of hemipteran pests, especially melon aphid, Aphis gossypii. The impact of low lethal concentrations of buprofezin on the biological parameters and expression profile of CHS1 gene were estimated for two successive generations of A. gossypii. The present result shows that the LC15 and LC30 of buprofezin significantly decreased the fecundity and longevity of both generations. Exposure of F0 individuals to both concentrations delay the developmental period in F1. Furthermore, the survival rate, intrinsic rate of increase (r), finite rate of increase (λ), and net reproductive rate (R0) were reduced significantly in progeny generation at both concentrations. However, the reduction in gross reproductive rate (GRR) was observed only at LC30. Although, the mean generation time (T) prolonged substantially at LC30. Additionally, expression of the CHS1 gene was significantly increased in F0 adults. Significant increase in the relative abundance of CHS1 mRNA transcript was also observed at the juvenile and adult stages of F1 generation following exposure to LC15 and LC30. Therefore, our results show that buprofezin could affect the biological traits by diminishing the chitin contents owing to the inhibition of chitin synthase activity in the succeeding generation of melon aphid.

Impact of the secondary plant metabolite Cucurbitacin B on the demographical traits of the melon aphid, Aphis gossypii.

Cucurbitacin B is a natural triterpene present in plants of Cucurbitaceae family, which are among the host plants for melon aphid, Aphis gossypii. In present study we characterized the effects of two cucurbitacin B concentrations on the biological parameters of adults (F0) and of juveniles and adults of their progeny (F1). The results showed that cucurbitacin B at 25 ppm significantly reduced the adult longevity and fecundity of both F0 and F1 generation. Exposure of F0 generation to 25 ppm though reduced the demographic traits of F1 including the intrinsic rate of increase r (day-1), generation time T (day), finite rate of increase λ (day-1), however, only net reproductive rate R0 (offspring/individual) decreased significantly. While 100 ppm reduced not only the longevity and fecundity of F0 generation but also the longevity of F1 generation. Fecundity of F1 was not affected by 100 ppm of cucurbitacin B, however, R0 (offspring/individual) and T (day) of F1 generation were lower than the control population. These results support the hypothesis that high contents of cucurbitacin B caused negative impact on melon aphid and could be used as a lead for classical selection of resistant varieties of plants that are main hosts for the melon aphid.