Microbe-mediated activation of indoxacarb in German cockroach (Blattella germanica L.).

The German cockroach (Blattella germanica L.) is a major urban pest worldwide and is notorious for its ability to detoxify and resist insecticides. German cockroaches have generalist feeding habits that expose them to a range of potential hazardous substances and host a wide variety of unique microbial species, which may potentially facilitate unique detoxification capabilities. Since field German cockroach populations are routinely exposed to both bait and spray insecticide treatments, we hypothesized whether these unique gut microbes might play roles in toxicological processes of the host insect. The goals of this research were to understand the metabolic processes inside the German cockroach gut after treatment with kanamycin, a broad-ranging antibiotic, and indoxacarb, an oxadiazine pro-insecticide used in cockroach bait products. In these experiments, two resistant cockroach strains were obtained from field populations in Danville, IL and compared to a susceptible laboratory strain that had no previous exposure to insecticides (J-wax). Roaches provided kanamycin-infused water had lower median mortality to indoxacarb compared to the control treatment in feeding bioassays regardless of strain, but in vial (surface contact) bioassays, only susceptible cockroaches experienced a shift in mortality apparently due to their greater susceptibility. When frass extracts of indoxacarb-fed cockroaches were analyzed, less of the active, hydrolytic metabolite DCJW (N-decarbomethoxyllated JW062) was produced relative to the parent compound indoxacarb with the antibiotic KAN. This result was further corroborated by hydrolase activity assays of whole homogenized cockroach guts. Taken together these results provide novel evidence of microbe-mediated pro-insecticide activation in the cockroach gut.

Differential microbial responses to antibiotic treatments by insecticide-resistant and susceptible cockroach strains (Blattella germanica L.).

The German cockroach (Blattella germanica L.) is a major urban pest worldwide and is known for its ability to resist insecticides. Past research has shown that gut bacteria in other insects can metabolize xenobiotics, allowing the host to develop resistance. The research presented here determined differences in gut microbial composition between insecticide-resistant and susceptible German cockroaches and compared microbiome changes with antibiotic treatment. Cockroaches received either control diet or diet plus kanamycin (KAN) to quantify shifts in microbial composition. Additionally, both resistant and susceptible strains were challenged with diets containing the insecticides abamectin and fipronil in the presence and absence of antibiotic. In both strains, KAN treatment reduced feeding, leading to higher doses of abamectin and fipronil being tolerated. However, LC50 resistance ratios between resistant and susceptible strains decreased by half with KAN treatment, suggesting gut bacteria mediate resistance. Next, whole guts were isolated, bacterial DNA extracted, and 16S MiSeq was performed. Unlike most bacterial taxa, Stenotrophomonas increased in abundance in only the kanamycin-treated resistant strain and was the most indicative genus in classifying between control and kanamycin-treated cockroach guts. These findings provide unique insights into how the gut microbiome responds to stress and disturbance, and important new insights into microbiome-mediated insecticide resistance.

Transcriptome Responses to Defined Insecticide Selection Pressures in the German Cockroach (Blattella germanica L.).

Cockroaches are important global urban pests from aesthetic and health perspectives. Insecticides represent the most cost-effective way to control cockroaches and limit their impacts on human health. However, cockroaches readily develop insecticide resistance, which can quickly limit efficacy of even the newest and most effective insecticide products. The goal of this research was to understand whole-body physiological responses in German cockroaches, at the metatranscriptome level, to defined insecticide selection pressures. We used the insecticide indoxacarb as the selecting insecticide, which is an important bait active ingredient for cockroach control. Six generations of selection with indoxacarb bait produced a strain with substantial (>20×) resistance relative to inbred control lines originating from the same parental stock. Metatranscriptome sequencing revealed 1,123 significantly differentially expressed (DE) genes in ≥two of three statistical models (81 upregulated and 1,042 downregulated; FDR P < 0.001; log2FC of ±1). Upregulated DE genes represented many detoxification enzyme families including cytochrome-P450 oxidative enzymes, hydrolases and glutathione-S-transferases. Interestingly, the majority of downregulated DE genes were from microbial and viral origins, indicating that selection for resistance is also associated with elimination of commensal, pathogenic and/or parasitic microbes. These microbial impacts could result from: (i) direct effects of indoxacarb, (ii) indirect effects of antimicrobial preservatives included in the selecting bait matrix, or (iii) selection for general stress response mechanisms that confer both xenobiotic resistance and immunity. These results provide novel physiological insights into insecticide resistance evolution and mechanisms, as well as novel insights into parallel fitness benefits associated with selection for insecticide resistance.