Mazarin Akami

The cowpea weevil, Callosobruchus maculatus (Fabricius) (Coleoptera: Bruchidae), is a major stored-product pest. Resistance to synthetic insecticides such as dichlorvos (DDVP) has prompted interest in alternatives with distinct modes of action. This study compared multigenerational transcriptional responses of C. maculatus to DDVP and the botanical insecticide Lippia adoensis (Hochst. ex Walp.) (Lamiales: Verbenaceae) essential oil (EO). DDVP exposure was associated with progressive fitness recovery across generations, together with upregulated expression of detoxification-related genes (e.g. Cyp6g1-2, Cyp6g1-3, Gst, Est2, Pod) and cholesterol metabolism-related genes (e.g. Dhcr24-1, Dhcr24-3). In contrast, L. adoensis EO was associated with consistently higher mortality and did not elicit comparable transcriptional changes in these gene sets. Clustering patterns placed EO-treated populations closer to controls, with less evidence of the DDVP-associated expression pattern. An antagonistic index comparing treatment-related fold changes indicated an opposing transcriptional relationship between EO and DDVP. Overall, the findings are consistent with L. adoensis EO acting without inducing the DDVP-associated detoxification expression pattern, suggesting a distinct transcriptional profile that may inform botanical-based approaches to pest management.

BACKGROUND: Substrate stiffness and internal pressure play key roles in mediating insect adaptability to mechanical cues through various mechanosensitive pathways. However, the involvement of mechanosensory perception in oviposition behavior of tephritid flies remains largely unexplored. Given that frugivorous tephritid flies use extraordinarily sharp‐tipped ovipositors to probe the stiff fruit peel before laying eggs, we speculated that mechanosensory‐responsive pathways mediate the oviposition behavior.RESULTS: In this study, we combined oviposition assays and RNA interference (RNAi) techniques to investigate how mechanosensory pathways mediate oviposition behaviors in the notorious invasive tephritid fly, Bactrocera dorsalis. The results show that both substrate stiffness and ovarian distention influenced the oviposition decision of female flies. Furthermore, silencing the BdTMC gene impaired the ability of flies to discriminate substrate hardness, while treatment with dsBdNomPC disrupted the internal pressure balance, leading to the accumulation of eggs in the ovary. Notably, a neuropeptide, BdNPLP1 was found to be involved in mechanosensitive signal transduction downstream of BdTMC and BdNomPC.CONCLUSIONS: Overall, our findings decipher the molecular mechanism underlying mechanosensory pathways mediating egg‐laying behavior in B. dorsalis. We propose that oviposition decision‐making is driven by a combination of internal and external mechanosensitive inputs, which offer new insights into the complex system of insect egg‐laying regulation. © 2025 Society of Chemical Industry.

Nitrogen is usually a restrictive nutrient that affects the growth and development of insects, especially of those living in low nitrogen nutrient niches. In response to the low nitrogen stress, insects have gradually developed symbiont-based stress response strategies—biological nitrogen fixation and nitrogenous waste recycling—to optimize dietary nitrogen intake. Based on the above two patterns, atmospheric nitrogen or nitrogenous waste (e.g., uric acid, urea) is converted into ammonia, which in turn is incorporated into the organism via the glutamine synthetase and glutamate synthase pathways. This review summarized the reaction mechanisms, conventional research methods and the various applications of biological nitrogen fixation and nitrogenous waste recycling strategies. Further, we compared the bio-reaction characteristics and conditions of two strategies, then proposed a model for nitrogen provisioning based on different strategies.

In order to understand the role of symbionts for their insect hosts, it is customary to treat them with antibiotics or to sterilize eggs (treatments), resulting in aposymbiotic and axenic insects, respectively. Such axenic insects can then be compared with untreated controls. Fruit flies often bear complex communities which are greatly reduced by such treatments. However, the bacterial community is not completely eliminated. Here, we examine the effect of these procedures on the structure of the remaining bacterial communities in Bactrocera dorsalis (Diptera: Tephritidae) and on the insect longevity. The antibiotics (Norfloxacin and Ceftazedime) were administered to 1-day-old adult flies through sugar meal for 7 days, and eggs were surface sterilized and dechorionated to produce axenic lines. The flies were starved of protein before they were offered full diets or diets containing non-essential amino acids only. Antibiotic and egg disinfection treatments resulted in a significant reduction of the vast majority of gut bacterial populations, especially Proteobacteria, Firmicutes and Bacteroidetes. On the other hand, antibiotic allowed the persistence of Actinobacteria, Cyanobacteria and Acidobacteria populations. In untreated control flies, longevity was extended irrespective of diet quality in comparison to treated flies. Conversely, when gut bacteria were largely reduced (aposymbiotic and axenic flies), longevity was reduced in the non-essential amino acids diet treatment versus slightly improved in the presence of a protein diet. We discuss these results in an ecological–life-history perspective.

Insects entertain intricate and mutualistic relationship with an array of microorganisms, which significantly influence their fitness, ecology and evolution. In recent decades, there has been increasing interest toward studying the effects of microbiome on many host insects (Dipterans, Lepidopterans, and Coleopterans). Studies so far realized indicate that gut microbiome contribute to host nutritional ecology, defense, immunity and lifespan. Bactrocera dorsalis (Tephritidae: Diptera) is a polyphagous fruit fly which attacks a huge variety of fruits and vegetables worldwide and has been placed as a quarantine species by many countries. To investigate the specific functions of the gut endosymbionts, it is a prerequisite to know the composition of gut bacterial communities whose manipulation will help to decipher their ecological relevance. Here, we used the culture-dependent technique to isolate and identify gut bacteria from B. dorsalis at different developmental stages. The results revealed 11 bacterial species from the third instar larvae, 18 and 12 from female and male populations, respectively. These bacteria were assigned to six families, namely, Enterobacteriaceae, Enterococcaceae, Staphylococcaceae, Streptococcaceae, Micrococcaceae and Bacillaceae. Bacterial species from these families were differentially represented in various samples, except Klebsiella oxytoca , Enterobacter cloacae, Pantoea dispers and Enterococcus faecalis that were detected at all developmental stages. Overall, Enterobacteriaceae was the most dominant family in females and third instar larvae accounting for 57.89% and 26.32%, respectively, while Enterococcaceae was dominant in males with 75% of the total bacterial taxa. These results suggest that B. dorsalis possesses a huge variety of cultivable bacteria that could be used to explore their specific functions on host physiology and fitness.

Bacillus thuringiensis (Bt) produce diverse insecticidal proteins to kill insect pests. Nevertheless, evolution of resistance to Bt toxins hampers the sustainable use of this technology. Previously, we identified down-regulation of a trypsin-like serine protease gene PxTryp_SPc1 in the midgut transcriptome and RNA-Seq data of a laboratory-selected Cry1Ac-resistant Plutella xylostella strain, SZ-R. We show here that reduced PxTryp_SPc1 expression significantly reduced caseinolytic and trypsin protease activities affecting Cry1Ac protoxin activation, thereby conferring higher resistance to Cry1Ac protoxin than activated toxin in SZ-R strain. Herein, the full-length cDNA sequence of PxTryp_SPc1 gene was cloned, and we found that it was mainly expressed in midgut tissue in all larval instars. Subsequently, we confirmed that the PxTryp_SPc1 gene was significantly decreased in SZ-R larval midgut and was further reduced when selected with high dose of Cry1Ac protoxin. Moreover, down-regulation of the PxTryp_SPc1 gene was genetically linked to resistance to Cry1Ac in the SZ-R strain. Finally, RNAi-mediated silencing of PxTryp_SPc1 gene expression decreased larval susceptibility to Cry1Ac protoxin in the susceptible DBM1Ac-S strain, supporting that low expression of PxTryp_SPc1 gene is involved in Cry1Ac resistance in P. xylostella. These findings contribute to understanding the role of midgut proteases in the mechanisms underlying insect resistance to Bt toxins.

The ability of a host plant to act as a substrate or media for larval development may depend on how good it is at offering suitable nutrients for bacterial growth. In this study, we hypothesized that the suitability of a fruit type for fruit fly larval development is positively correlated with the ability of that fruit to act as a substrate/media for fruit fly symbiotic bacterial growth. We allowed a single female fruit fly to lay eggs on five different host fruits, then we monitored the larval development parameters across five generations and analyzed the bacterial community structure of larvae developing in 2 of these hosts (apple and banana) at the first and fifth generations. Results indicate that the larval length and dry weight did not vary significantly across experimental generations, but were greatly affected by fruit types and larval stages. The larval development time was extended considerably in apple and tomato but shortened in banana and mango. There was a significant shift in bacterial community structure and composition across fruits and generations. The bacterial community of larvae within the same fruit (apple and banana) clustered and was similar to the parental female (with the predominance of Proteobacteria), but there was a shift at the fifth generation (dominance of Firmicutes). Banana offered a suitable better development and growth to larvae and bacteria, respectively, compared to apple in which reduced larval development and bacterial growth were recorded. Although additional experiments are needed to adequately show that the differences in microbiome seen in fruit fly larval guts are the actual driver of different developmental outcomes of larvae on the different fruits, at the very least, our study has provided intriguing data suggesting interaction between the diets and gut microbial communities on insect development. Importance and Significance of the study: Tephritid fruit flies entertain complex interactions with gut bacteria. These bacteria are known to provide nutritional benefits to their hosts, by supplementing missing nutrients from the host diets and regulating energy balance. Foraging for food is a risky exercise for the insect which is exposed to ecological adversities, including predators. Therefore, making beneficial choice among available food substrates is a question of survival for the flies and bacteria as well. Our study demonstrates interactions between the host fly and its intestinal bacteria in sustaining the larval development while foraging optimally on different fruit types. These findings add a novel step into our understanding of the interactions between the gut microbial communities and B. dorsalis and provide avenues for developing control strategies to limit the devastative incidence of the fly.

Background The gut bacteria of tephritid fruit flies play prominent roles in nutrition, reproduction, maintenance and ecological adaptations of the host. Here, we adopted an approach based on direct observation of symbiotic or axenic flies feeding on dishes seeded with drops of full diet (containing all amino acids) or full diet supplemented with bacteria at similar concentrations to explore the effects of intestinal bacteria on foraging decision and fitness of Bactrocera dorsalis. Results The results show that intestinal probiotics elicit beneficial foraging decision and enhance the female reproduction fitness and survival of B. dorsalis (symbiotic and axenic), yet preferences for probiotic diets were significantly higher in axenic flies to which they responded faster compared to full diet. Moreover, females fed diet supplemented with Pantoea dispersa and Enterobacter cloacae laid more eggs but had shorter lifespan while female fed Enterococcus faecalis and Klebsiella oxytoca enriched diets lived longer but had lower fecundity compared to the positive control. Conversely, flies fed sugar diet (negative control) were not able to produce eggs, but lived longer than those from the positive control. Conclusions These results suggest that intestinal bacteria can drive the foraging decision in a way which promotes the reproduction and survival of B. dorsalis. Our data highlight the potentials of gut bacterial isolates to control the foraging behavior of the fly and c empower the sterile insect technique (SIT) program through the mass rearing.

BACKGROUND: Pair‐rule genes were identified and named for their role in segmentation in animal embryos. Paired, a homolog of mammalian PAX3, acts as one of several pair‐rule genes and is key in defining the boundaries of future parasegments and segments during insect embryogenesis.RESULTS: We cloned the paired gene from the oriental fruit fly, Bactrocera dorsalis, and then applied CRISPR/Cas9‐mediated genome editing to investigate its physiological function in the embryonic stage of this pest. We identified one transcript for a paired homolog in B. dorsalis, which encodes a protein containing a Paired Box domain and a Homeobox domain. Phylogenetic analysis confirmed that the paired gene is highly conserved and the gene was highly expressed at the 12–14 h‐old embryonic stage. Knock‐out of Bdpaired led to lack of segment boundaries, cuticular deficiency, and embryonic lethality. Sequence analysis of the CRISPR/Cas9 mutants exhibited different insertion and deletions in the Bdpaired locus. In addition, the relative expression of Wingless (Wg) and Abdominal A (Abd‐A) genes were significantly down‐regulated in the Bdpaired mutant embryos.CONCLUSION: These results indicate that Bdpaired gene is critical for the embryonic development of B. dorsalis, and could be a novel molecular target for genetic‐based pest management practices to combat this serious invasive pest. © 2019 Society of Chemical Industry