Level of Pyrethroid-resistance Associated with Cytochrome P450 Expression in German Cockroach Blattella germanica (Blattodea: Ectobiidae) in the Field Collected Strains

Background: Cytochrome P450-dependent monooxygenases are a very important metabolic system involve in insecti­cide resistance. This study was conducted to find the association between the expression level of cytochrome P450 (CYP450) and permethrin-resistance level among four strains of the German cockroach Blattella germanica (L) (Blat­todea: Ectobiidae). Methods: Three field strains of German cockroach with different frequency of exposure to pesticides, and a laboratory susceptible strain were used in the present study. Insecticide susceptibility bioassays were carried out to detect re­sistance to permethrin. The concentration of CYP450 in each strain was determined using ion-exchange HPLC chroma­tography. Biochemical assays was performed to analyse CYP450 activities. Results: The resistance ratios (RR) to permethrin among three field strains were 3.29, 4.10 and 6.17-fold comping with the susceptible stain. The CYP450 activity of three field strains was 1.6, 2.4 and 2.7 times higher than in the sus­ceptible stain. The amount of CYP450 per mg of protein was significantly different between the susceptible and the three re­sistant strains. The resistant cockroaches showed a relatively high expression of CYP450 enzymes. A strong corre­la­tion was found between permethrin resistance level and total concentration of CYP450 enzymes. Conclusion: The results of current study show that more frequent usage of a pyrethroid insecticide cause the metabolic insecticide resistance to rise in German cockroach. Therefore, there is a ratio correlation between resistance level and monooxygenases activities in insect. Thus, the control program must be managed according to history of pesticide us­age.

The effectiveness of commercial gel baits against German cockroach Blattella germanica (Linnaeus, 1767) in Indonesia

Gel baits product sold commercially to the public has long been used to control German cockroach populations in urban areas. The effectiveness and resistance level of gel bait products need to be known for the successful control of German cockroaches in the field. However, there have been no reports of resistance in Indonesia. This study used toxicity tests of six gel bait products (MF, CBG, SPB, OTG, BTX, and UBA) against two populations of adult male German cockroaches (VCRU-WHO and RKMN-BKT). Fipronil, boric acid and emamectin benzoate containing gel baits were effective to knock down and kill the German cockroaches. In contrast, gel bait with imidacloprid as an active ingredient was only useful to knock down the German cockroaches. Commercial gel bait product in Indonesia are still useful in controlling German cockroaches’ populations with a time of effectiveness to cause mortality between three to four weeks after being exposed in the field and the level of cockroach resistance to gel bait are susceptible until low resistance.

Interkingdom Gut Microbiome and Resistome of the Cockroach Blattella germanica

ABSTRACT Cockroaches are intriguing animals with two coexisting symbiotic systems, an endosymbiont in the fat body, involved in nitrogen metabolism, and a gut microbiome whose diversity, complexity, role, and developmental dynamics have not been fully elucidated. In this work, we present a metagenomic approach to study Blattella germanica populations not treated, treated with kanamycin, and recovered after treatment, both naturally and by adding feces to the diet, with the aim of better understanding the structure and function of its gut microbiome along the development as well as the characterization of its resistome. IMPORTANCE For the first time, we analyze the interkingdom hindgut microbiome of this species, including bacteria, fungi, archaea, and viruses. Network analysis reveals putative cooperation between core bacteria that could be key for ecosystem equilibrium. We also show how antibiotic treatments alter microbiota diversity and function, while both features are restored after one untreated generation. Combining data from B. germanica treated with three antibiotics, we have characterized this species’ resistome. It includes genes involved in resistance to several broad-spectrum antibiotics frequently used in the clinic. The presence of genetic elements involved in DNA mobilization indicates that they can be transferred among microbiota partners. Therefore, cockroaches can be considered reservoirs of antibiotic resistance genes (ARGs) and potential transmission vectors.

Salivary Digestion Extends the Range of Sugar-Aversions in the German Cockroach

Saliva has diverse functions in feeding behavior of animals. However, the impact of salivary digestion of food on insect gustatory information processing is poorly documented. Glucose-aversion (GA) in the German cockroach, Blattella germanica, is a highly adaptive heritable behavioral resistance trait that protects the cockroach from ingesting glucose-containing-insecticide-baits. In this study, we confirmed that GA cockroaches rejected glucose, but they accepted oligosaccharides. However, whereas wild-type cockroaches that accepted glucose also satiated on oligosaccharides, GA cockroaches ceased ingesting the oligosaccharides within seconds, resulting in significantly lower consumption. We hypothesized that saliva might hydrolyze oligosaccharides, releasing glucose and terminating feeding. By mixing artificially collected cockroach saliva with various oligosaccharides, we demonstrated oligosaccharide-aversion in GA cockroaches. Acarbose, an alpha-glucosidase inhibitor, prevented the accumulation of glucose and rescued the phagostimulatory response and ingestion of oligosaccharides. Our results indicate that pre-oral and oral hydrolysis of oligosaccharides by salivary alpha-glucosidases released glucose, which was then processed by the gustatory system of GA cockroaches as a deterrent and caused the rejection of food. We suggest that the genetic mechanism of glucose-aversion support an extended aversion phenotype that includes glucose-containing oligosaccharides. Salivary digestion protects the cockroach from ingesting toxic chemicals and thus could support the rapid evolution of behavioral and physiological resistance in cockroach populations.

Modulation of fatty acid elongation generates sexually dimorphic hydrocarbons and female attractiveness in Blattella germanica (L.)

Insect cuticular compounds serve multiple functions. As important intersexual signaling chemicals, they show variation between the sexes, but little is known about the underlying molecular mechanisms. Here, we report that sexually dimorphic hydrocarbons (SDHCs) are generated by a fatty acid elongase gene that is regulated by sex-differentiation genes in the German cockroach, Blattella germanica. Sexually mature females possess more C29 cuticular hydrocarbons (CHCs), especially the contact sex pheromone precursor 3,11-DimeC29. An RNAi screen and heterologous expression revealed that BgElo12 and BgElo24 were involved in HC production, but only BgElo12 was responsible for SDHCs. Repressing female-enriched BgElo12 masculinized the female CHC profile, decreased contact sex pheromone level, and reduced the female sexual attractiveness. Moreover, RNAi of the sex-differentiation genes BgTra or BgDsx modulated both BgElo12 transcripts and CHC profiles in females and males. The SDHCs are shaped by sexual selection, as females use them to keep high levels of sex pheromone.

Salivary Digestion Extends the Range of Sugar-Aversions in the German Cockroach

Saliva has diverse functions in feeding behavior of animals. However, the impact of salivary digestion of food on insect gustatory information processing is poorly documented. Glucose-aversion (GA) in the German cockroach, Blattella germanica, is a highly adaptive heritable behavioral resistance trait that protects the cockroach from ingesting glucose-containing-insecticide-baits. In this study, we confirmed that GA cockroaches rejected glucose, but they accepted oligosaccharides. However, whereas wild-type cockroaches that accepted glucose also satiated on oligosaccharides, GA cockroaches ceased ingesting the oligosaccharides within seconds, resulting in significantly lower consumption. We hypothesized that saliva might hydrolyze oligosaccharides, releasing glucose and terminating feeding. By mixing artificially collected cockroach saliva with various oligosaccharides, we demonstrated oligosaccharide-aversion in GA cockroaches. Acarbose, an alpha-glucosidase inhibitor, prevented the accumulation of glucose and rescued the phagostimulatory response and ingestion of oligosaccharides. Our results indicate that pre-oral and oral hydrolysis of oligosaccharides by salivary alpha-glucosidases released glucose, which was then processed by the gustatory system of GA cockroaches as a deterrent and caused the rejection of food. We suggest that the genetic mechanism of glucose-aversion support an extended aversion phenotype that includes glucose-containing oligosaccharides. Salivary digestion protects the cockroach from ingesting toxic chemicals and thus could support the rapid evolution of behavioral and physiological resistance in cockroach populations.