Toxicity of isocycloseram, an isoxazoline insecticide, against laboratory and field-collected German cockroaches (Blattodea: Ectobiidae).

Isocycloseram is a new insecticide in the isoxazoline class that targets insect GABA-gated chloride channels. In this study, we evaluated a cockroach gel bait formulation containing 1% isocycloseram against a susceptible strain (UCR) and 5 field-collected strains (WM, RG386, Ryan, CDR, and SY) of the German cockroach, Blattella germanica (L.) (Blattodea: Ectobiidae), and compared it with several commercial insecticide baits in the laboratory. Using the Ebeling choice box method, we also tested a residual deposit of an SC formulation of isocycloseram against the UCR, RG386, and Ryan strains. The isocycloseram bait was among the fastest-performing treatments against adult males (mean survival time: 0.9-2.7 days) and mixed stages and sexes (mean survival time: 1.4-5.4 days) across all strains. Secondary transfer effects of the bait were demonstrated in the UCR strain by exposing new adult males to individuals killed by direct bait treatment. Physiological resistance was not detected in the WM, CDR, and RG386 strains with topical treatment of a diagnostic dose (3× LD95) of isocycloseram developed using the UCR strain. However, topical assays revealed resistance ratios (RR50) of 1.6 and 3.0× in the Ryan and SY strains, respectively. The performance of a 0.05% isocycloseram residual application against the Ryan strain was improved with the addition of piperonyl butoxide.

Overexpression of cytochrome P450 gene CYP6K1 is associated with pyrethroid resistance in German cockroaches (Blattodea: Ectobiidae) from California.

We investigated the expression of 4 cytochrome P450 genes (CYP4G19, CYP6J1, CYP6K1, and CYP4C21) in 4 field-collected strains (WM, RG386, CDR, and Ryan) of the German cockroach, Blattella germanica (L.), collected from California. The UCR susceptible strain was used as a comparison. Topical assays using a diagnostic dose (3× LD95) of deltamethrin revealed decreased sensitivity in all field-collected strains with mortality ranging from 0% to 58%, and the addition of PBO before deltamethrin treatment increased mortality to 52.5%-87.5%. Using qPCR to investigate the expression levels of CYP4G19, CYP4C21, CYP6J1, and CYP6K1, we found that only CYP6K1 was significantly overexpressed (2.1-5.8× higher) in all field-collected strains when compared to the UCR strain. Next, we investigated the role of the CYP6K1 gene by performing gene knockdown using RNAi. After dsCYP6K1 treatment, the expression levels of CYP6K1 in WM and Ryan strains were significantly reduced (P < 0.01) by 91%-94% vs. those treated with dsEGFP (control) on the third and sixth day posttreatment. RG386, CDR, and Ryan strains were more susceptible compared to their respective controls to topically applied deltamethrin 6 days after treatment with dsCYP6K1. This study provides evidence of the involvement of the P450 CYP6K1 gene in pyrethroid resistance in some populations of German cockroaches.

Oral toxicity of an artificial sweetener sucralose on the German cockroach (Blattodea: Ectobiidae) and its impact on water balance and gut microbiome.

Artificial or non-nutritive sweeteners are indigestible by most animals. Some sweeteners are orally toxic to insects and have received recent interest as potential safe insecticides due to their low mammalian toxicity. In this study, we investigated the oral toxicity of sucralose on insecticide-susceptible and resistant German cockroaches, Blattella germanica (L.). In a nonchoice test, we evaluated 5, 10, and 20% sucralose solutions. Depending on the cockroach strains, mean mortality ranged from 62.5 to 92.5%, 15 to 55%, and 2.5 to 27.5% for 20, 10, and 5% sucralose, respectively. Next, we measured the impact of a 20% sucralose treatment on water loss rates in the cockroach strains. All strains lost 23.0-30.29% of body water by 6 d. Dehydrated cockroaches were more prone to be killed by sucralose than nondehydrated ones. Lastly, we evaluated the effect of 20% sucralose treatment on gut bacterial composition and found the diversity of gut bacteria in treated cockroaches was significantly reduced after 3 days, implicating a rapid change in the alimentary environment.

Insecticide resistance and its potential mechanisms in field-collected German cockroaches (Blattodea: Ectobiidae) from Thailand.

We investigated insecticide resistance profiles of field populations of the German cockroach, Blattella germanica (L.), collected from central regions of Thailand. Seven strains (PW, RB, MTH, MTS, TL, AY, and SP) were evaluated with diagnostic doses (DD; 3 × LD95 generated from a susceptible strain) of deltamethrin, fipronil, and imidacloprid using topical assays and compared with a susceptible strain (DMSC). Results showed fipronil (2-27% mortality), deltamethrin (16-58% mortality), and imidacloprid (15-75% mortality) resistance in the field strains. Synergism studies with piperonyl butoxide (PBO) and S,S,S-tributyl phosphorotrithioate (DEF) in combination with the DD of insecticides significantly increased (P < 0.05) mortality of the test insects of the field strains suggesting the involvement of P450 monooxygenase and esterase pathways of detoxification. Gel bait evaluations demonstrated that all field-collected strains were resistant to Maxforce Forte (0.05% fipronil), Maxforce Fusion (2.15% imidacloprid), and Advion Cockroach Gel Bait (0.6% indoxacarb) with mean survival times ranging from 1.87-8.27, 1.77-11.72, and 1.19-3.56 days, respectively. Molecular detection revealed that the Rdl mutation was completely homozygous in all field-collected strains except in the PW strain. Field-collected strains were screened for 3 voltage-gated sodium channel (VGSC) mutations associated with pyrethroid resistance. The L993F mutation was present in 5 strains, but no C764R and E434K mutations were detected.

Combined metabolic and target-site resistance mechanisms confer fipronil and deltamethrin resistance in field-collected German cockroaches (Blattodea: Ectobiidae).

Despite insecticide resistance issues, pyrethroids and fipronil have continued to be used extensively to control the German cockroach, Blattella germanica (L.) (Blattodea: Ectobiidae) for more than two decades. We evaluated the physiological insecticide resistance in five German cockroach populations collected from 2018 to 2020 and measured the extent of metabolic detoxification and target-site insensitivity resistance mechanisms. Topically applied doses of the 3 x LD95 of deltamethrin, fipronil, DDT, or dieldrin of a susceptible strain (UCR, Diagnostic Dose) failed to cause >23% mortality, and the 10 x LD95 of deltamethrin or fipronil failed to cause >53% mortality. All field-collected strains possessed a combination of metabolic and target-site insensitivity mechanisms that cause reduced susceptibility. Elevated activities of esterase and glutathione S-transferase were measured, and the synergists piperonyl butoxide or S,S,S-tributyl phosphorotrithioate increased topical mortality up to 100% for deltamethrin and 93% for fipronil 10 x LD95. The target-site mutations L993F of the para-homologous sodium channel and A302S of the GABA-gated chloride channel associated with pyrethroid and fipronil resistance, respectively, were found at ~80-100% frequency in field populations. Pyrethroid and fipronil spray formulations also were ineffective in a choice box assay against field-collected strains suggesting that these treatments would fail to control cockroaches under field conditions.

Reduced Susceptibility Towards Commercial Bait Insecticides in Field German Cockroach (Blattodea: Ectobiidae) Populations From California.

Gel bait insecticides have been extensively used to manage the German cockroach, Blattella germanica (L.) (Blattodea: Ectobiidae), but issues with reduced effectiveness of such formulations are becoming increasingly common. We collected five field strains of German cockroaches in California and evaluated them against five commercial bait products [Maxforce FC Magnum (0.05% fipronil), Maxforce Impact (1% clothianidin), Advion Evolution (0.6% indoxacarb), Optigard (0.1% emamectin benzoate) and Siege (2% hydramethylnon)]. Increased survivorship and incomplete mortality towards all baits were recorded in the field strains. We assessed susceptibility to the active ingredients fipronil, clothianidin, indoxacarb, abamectin, hydramethylnon, and deltamethrin using topical bioassays with diagnostic doses (3 × LD95 and 10 × LD95) developed from the UCR susceptible strain. Low mortality was registered when tested with the 3 × LD95's of deltamethrin (0%), fipronil (0-3%), and clothianidin (13-27%); low to moderate mortality when treated with the 3 × LD95 of indoxacarb (13-63%), and moderate to high mortality after treatment with the 3 × LD95 of abamectin (80-100%) and hydramethylnon (70-83%). The mortality of all strains remained low after treatment with the 10 × LD99 of deltamethrin (0-20%) and low to moderate with fipronil (20-70%). We found negative correlations (P < 0.05) between Advion Evolution mean survival time and indoxacarb 10 × LD95 mortality and between Maxforce Impact and clothianidin 10 × LD95 mortality. These findings demonstrate multiple resistance towards all tested commercial bait insecticides except Optigard, suggesting the effectiveness of avermectin products in resistance management programs.

The Impact of Artificial Sweeteners on Insects.

Artificial sweeteners are sweet-tasting additives found in consumable products as substitutes for naturally occurring sugars. They are derived from plant extracts or manufactured by chemical synthesis. Ingestion of sweeteners by insects can lead to significant physiological effects, such as mortality, decreased fecundity, and behavioral change. Due to their low toxicity toward humans and the issues associated with conventional insecticide usage, artificial sweeteners have recently gained attention for their potential use as biorational insecticides. Here, we review their impact on insects and potential as novel insecticides.

Verification of Argentine ant defensive compounds and their behavioral effects on heterospecific competitors and conspecific nestmates.

The invasive Argentine ant (Linepithema humile) has become established worldwide in regions with Mediterranean or subtropical climates. The species typically disrupts the balance of natural ecosystems by competitively displacing some native ant species via strong exploitation and interference competition. Here we report that Argentine ants utilize glandular secretions for inter and intra-specific communications during aggressive interactions with a heterospecific competitor, California harvester ant (Pogonomyrmex californicus). Chemical analyses indicated that Argentine ants deploy glandular secretions containing two major volatile iridoids, dolichodial and iridomyrmecin, on the competitor's cuticular surface during aggressive interactions. Bioassays indicated that the glandular secretions function as a defensive allomone, causing high levels of irritation in the heterospecific. Furthermore, the same glandular secretions elicited alarm and attraction of conspecific nestmates, potentially enabling more rapid/coordinated defense by the Argentine ants. Two major volatile constituents of the glandular secretion, dolichodial and iridomyrmecin, were sufficient to elicit these responses in conspecifics (as a mixture or individual compounds). The current study suggests that invasive Argentine ants' superior exploitation and interference competition may rely on the species' effective semiochemical parsimony.