Distinct roles of two RDL GABA receptors in fipronil action in the diamondback moth (Plutella xylostella)

The phenylpyrazole insecticide fipronil blocks resistance to dieldrin (RDL) γ-aminobutyric acid (GABA) receptors in insects, thereby impairing inhibitory neurotransmission. Some insect species, such as the diamondback moth (Plutella xylostella), possess more than one Rdl gene. The involvement of multiple Rdls in fipronil toxicity and resistance remains largely unknown. In this study, we investigated the roles of two Rdl genes, PxRdl1 and PxRdl2, in P. xylostella fipronil action. In Xenopus oocytes, PxRDL2 receptors were 40 times less sensitive to fipronil than PxRDL1. PxRDL2 receptors were also less sensitive to GABA compared with PxRDL1. Knockout of the fipronil-sensitive PxRdl1 reduced the fipronil potency 10-fold, whereas knockout of the fipronil-resistant PxRdl2 enhanced the fipronil potency 4.4-fold. Furthermore, in two fipronil-resistant diamondback moth field populations, PxRdl2 expression was elevated 3.7- and 4.1-fold compared with a susceptible strain, whereas PxRdl1 expression was comparable among the resistant and susceptible strains. Collectively, our results indicate antagonistic effects of PxRDL1 and PxRDL2 on fipronil action in vivo and suggest that enhanced expression of fipronil-resistant PxRdl2 is potentially a new mechanism of fipronil resistance in insects.     

Resistance to fipronil in Drosophila simulans: influence of two point mutations in the RDL GABA receptor subunit

The Eyguieres 42 strain of Drosophila simulans, obtained by laboratory selection, displayed approximately 20,000-fold resistance to the insecticide fipronil. Molecular cloning of the cDNA encoding the RDL GABA receptor subunit of this strain revealed the presence of two mutations: the Rdl mutation (A301G) and an additional mutation in the third transmembrane domain (T350M). In order to assess the individual and combined roles of the two mutations in fipronil resistance, the functional properties of wild-type, A301G, T350M and A301G/T350M homomultimeric RDL receptors were compared by expression in Xenopus oocytes. In wild-type receptors, the inhibition of GABA (EC(30))-induced currents by fipronil and picrotoxin was enhanced by repeated GABA applications. The A301G mutation nearly abolished this effect, decreased the sensitivity to fipronil and picrotoxin and increased the reversibility of inhibition. The T350M mutation also reduced the sensitivity to both antagonists. Of the four receptor variants tested, the double mutant showed the highest resistance to fipronil, following repeated GABA applications. In conclusion, the present study emphasizes new aspects of the pharmacological alterations induced by the Rdl mutation and shows that resistance to GABA receptor-directed insecticides may implicate a mutation distinct from Rdl.     

Quantification of fipronil and its metabolite fipronil sulfone in rat plasma over a wide range of concentrations by LC/UV/MS

Fipronil is an insecticide extensively used to treat pets, which has been identified as a potential thyroid disruptor in the rat. In this species, fipronil is mainly metabolized to fipronil sulfone and plasma concentrations of fipronil sulfone can be at least 20-fold higher than those of fipronil. Investigations of fipronil and fipronil sulfone exposure in blood remain sparse because of the lack of convenient and suitable analytical methods. We have developed and validated an LC/UV/MS/MS method to quantify both fipronil and fipronil sulfone within a wide range of concentrations in rat plasma. The double detection UV and MS coupled on-line enabled the concentrations to be measured over a 3 Log range (2.5–2500 ng/mL). The volume of sample required for the extraction by solid phase extraction was reduced to 75 μL with a recovery higher than 70%. The two-detection method agreement, evaluated with a Bland–Altman plot, was good for concentrations between 50 and 150 ng/mL. The method was applied to monitor plasma concentrations following a commonly used dosage regimen for the toxicological evaluation of fipronil in rats.     

Evaluation of flubendiamide‐induced mitochondrial dysfunction and metabolic changes in Helicoverpa armigera (Hubner)

Phthalic acid diamide insecticides are the most effective insecticides used against most of the lepidopteran pests including Helicoverpa armigera, a polyphagous pest posing threat to several crops worldwide. The present studies were undertaken to understand different target sites and their interaction with insect ryanodine receptors (RyR). Bioassays indicated that flubendiamide inhibited the larval growth in dose‐dependent manner with LD₅₀ value of 0.72 μM, and at 0.8 μM larval growth decreased by about 88%. Flubendiamide accelerated the Ca²⁺‐ATPase activity in dose‐dependent trend, and at 0.8 μM, the activity was increased by 77.47%. Flubendiamide impedes mitochondrial function by interfering with complex I and F₀F₁‐ATPase activity, and at 0.8 μM the inhibition was found to be about 92% and 50%, respectively. In vitro incubation of larval mitochondria with flubendiamide induced the efflux of cytochrome c, indicating the mitochondrial toxicity of the insecticide. Flubendiamide inhibited lactate dehydrogenase and the accumulation of H₂O₂, thereby preventing the cells from lipid peroxidation compared to control larvae. At 0.8 μM the LDH, H₂O₂ content and lipid peroxidation was inhibited by 98.44, 70.81, and 70.81%, respectively. Cytochrome P450, general esterases, AChE, and antioxidant enzymes (catalase and superoxide dismutase) exhibited a dose‐dependent increasing trend, whereas alkaline phosphatase and the midgut proteases, except amino peptidase, exhibited dose‐dependent inhibition in insecticide‐fed larvae. The results suggest that flubendiamide induced the harmful effects on the growth and development of H. armigera larvae by inducing mitochondrial dysfunction and inhibition of midgut proteases, along with its interaction with RyR.     

氟虫腈、吡虫啉作为黑翅土白蚁诱杀药剂的效果

毒力测定结果表明,0.025~0.4μg/mL氟虫腈和吡虫啉分别在药后3 d和5 d对黑翅土白蚁Odontotermes formosanus表现出明显的毒杀效果,氟虫腈和吡虫啉药后1 d的LC50分别为药后5 d的509倍和63.8倍,2种药剂对黑翅土白蚁的毒杀效果均比较缓慢。毒性传递试验表明,0.5μg/g毒沙处理白蚁1 h后,氟虫腈和吡虫啉的致死毒性均可被传毒白蚁传递给受毒白蚁。驱避作用试验表明,50μg/mL氟虫腈对黑翅土白蚁无明显的驱避作用,而50μg/mL吡虫啉对黑翅土白蚁表现出了明显的驱避作用。可见,2种供试药剂中,氟虫腈是较理想的白蚁诱杀药剂。     

Glutamatergic and GABAergic effects of fipronil on olfactory learning and memory in the honeybee

We investigated here the role of transmissions mediated by GABA and glutamate-gated chloride channels (GluCls) in olfactory learning and memory in honeybees, both of these channels being a target for fipronil. To do so, we combined olfactory conditioning with injections of either the GABA- and glutamate-interfering fipronil alone, or in combination with the blocker of glutamate transporter l-trans-Pyrrolidine-2,4-Dicarboxylicacid (l -trans-PDC), or the GABA analog Trans-4-Aminocrotonic Acid (TACA). Our results show that a low dose of fipronil (0.1 ng/bee) impaired olfactory memory, while a higher dose (0.5 ng/bee) had no effect. The detrimental effect induced by the low dose of fipronil was rescued by the coinjection of l-trans-PDC but was rather increased by the coinjection of TACA. Moreover, using whole-cell patch-clamp recordings, we observed that l-trans-PDC reduced glutamate-induced chloride currents in antennal lobe cells. We interpret these results as reflecting the involvement of both GluCl and GABA receptors in the impairment of olfactory memory induced by fipronil.     

Butylene fipronil induces apoptosis in PC12 murine nervous cells via activation of p16‐CDK4/6‐cyclin D1 and mitochondrial apoptotic pathway

Butylene fipronil (BFPN) is a phenylpyrazole insecticide, acting at the γ‐aminobutyric acid (GABA) receptor. Here, we show that BFPN inducedcytotoxicity in PC12 murinenervous cells, which lacks GABA receptor. Treatment with BFPN for 48 hours significantly enhanced G0/G1 arrest and induced apoptosis. BFPN decreased the expression of cyclin‐dependent kinase (CDK4 and CDK6) and increased P16 and cyclin D1. Simultaneously, Bcl‐2 protein was declined while Bax and cytochrome c were significantly enhanced in BFPN‐treated groups. The apoptotic enzymes caspase‐8, ‐9, and ‐3 were also activated by BFPN. Furthermore, treatment with BFPN significantly stimulated reactive oxygen species (ROS) generation, and pretreatment with antioxidant diphenyleneiodonium, substantially reduced cell death. Overall, these results suggest that BFPN is effective to induce G0/G1‐phase arrest and apoptosis in PC12 murine nervous cell. Stimulating ROS generation and activation of P16‐CDK4/6‐cyclin D1 and mitochondrial apoptotic pathway may participate in the cytotoxicity of BFPN.     

Fipronil recognition by the FA1 site of human serum albumin

Fipronil is a broad‐spectrum pesticide widely used in agriculture, horticulture, and forestry. Because fipronil can cause a variety of toxic effects in animals and humans, its use is authorized as a pesticide in veterinary medicinal products for pets, but not for the treatment of livestock animals whose products are intended for consumption. Recently, however, the presence of fipronil residues has been detected in the eggs and meat of layer hens from farms located in different European countries. Given the relevance of fipronil toxicity for human health, it is important to gain information concerning its fate in the human body, including its binding mode to human serum albumin (HSA), the most abundant protein in plasma. Here, the inhibition of heme‐Fe(III) binding to the fatty acid site 1 (FA1) of HSA by fipronil is reported. Docking simulations support functional data, indicating that the FA1 site is the preferential cleft for fipronil recognition by HSA. The affinity of fipronil for HSA (K_f = 1.9 × 10^?6 M, at pH 7.3, and 20.0°C) may be relevant in vivo. Indeed, HSA could play a pivotal role in fipronil transport and scavenging, thus reducing the pesticide‐free plasmatic levels, with consequent reduced systemic toxicity. In turn, fipronil binding to the FA1 site of HSA could impair the recognition of endogenous and exogenous molecules.     

Inheritance mode and properties of fipronil resistance in Oxycarenus hyalinipennis Costa (Hemiptera: Lygaeidae)

The dusky cotton bug (DCB), Oxycarenus hyalinipennis Costa (Hemiptera: Lygaeidae) is one of the most important pests of cotton. Fipronil belongs to phenylpyrazole and is being used against various insect pests. To reduce the resistance to this insecticide, the present study was planned to examine the inheritance mode and preliminary mechanism of resistance to fipronil in DCB. A fipronil selected strain had a 9855-fold level of resistance after 11 generations of selection with fipronil. The median lethal concentration of reciprocal crosses F₁ (Fipro-Sel.♂×Lab-Pop♀) and F₁^-(Fipro-Sel.♀× Lab-Pop♂) showed no significant difference and degree of dominance values were 0.60 and 0.58 for F1 and F1^-), respectively, suggesting autosomal and incompletely dominance of resistance to fipronil in fipronil selected strain of DCB. Monogenic model of inheritance revealed polygenic resistance to fipronil. Realized heritability (h²) value was 0.24 for fipronil resistance. The study of synergism indicated that PBO (Piperonyl butoxide) and DEF (S, S-butyl phosphorotrithioateases) did not enhance the toxicity of fipronil on both populations.The study on inheritance mode of fipronil resistance in DCB may help delay the resistance with insecticides.     

In vitro metabolism of fipronil by human and rat cytochrome P450 and its interactions with testosterone and diazepam

Fipronil (5-amino-1-[2,6-dichloro-4-(trifluoromethyl)phenyl]-4-[(trifluoromethyl)sulfinyl]-1H-pyrazole-3-carbonitrile) is a highly active, broad spectrum insecticide from the phenyl pyrazole family, which targets the gamma-amino butyric acid (GABA) receptor. Although fipronil is presently widely used as an insecticide and acaricide, little information is available with respect to its metabolic fate and disposition in mammals. This study was designed to investigate the in vitro human metabolism of fipronil and to examine possible metabolic interactions that fipronil may have with other substrates. Fipronil was incubated with human liver microsomes (HLM) and several recombinant cytochrome P450 (CYP) isoforms obtained from BD Biosciences. HPLC was used for metabolite identification and quantification. Fipronil sulfone was the predominant metabolite via CYP oxidation. The K(m) and V(max) values for human liver microsomes are 27.2 microM and 0.11 nmol/mg proteinmin, respectively; for rat liver microsomes (RLM) the K(m) and V(max) are 19.9 microM and 0.39 nmol/mg proteinmin, respectively. CYP3A4 is the major isoform responsible for fipronil oxidation in humans while CYP2C19 is considerably less active. Other human CYP isoforms have minimal or no activity toward fipronil. Co-expression of cytochrome b(5) (b(5)) is essential for CYP3A4 to manifest high activity toward fipronil. Ketoconazole, a specific inhibitor of CYP3A4, inhibits 78% of the HLM activity toward fipronil at a concentration of 2 microM. Oxidative activity toward fipronil in 19 single-donor HLMs correlated well with their ability to oxidize testosterone. The interactions of fipronil and other CYP3A4 substrates, such as testosterone and diazepam, were also investigated. Fipronil metabolism was activated by testosterone in HLM but not in CYP3A4 Supersomes. Testosterone 6beta-hydroxylation in HLM was inhibited by fipronil. Fipronil inhibited diazepam demethylation but had little effect on diazepam hydroxylation. The results suggest that fipronil has the potential to interact with a wide range of xenobiotics or endogenous chemicals that are CYP3A4 substrates and that fipronil may be a useful substrate for the characterization of CYP3A4 in HLM.     

Photo-controlled release of fipronil from a coumarin triggered precursor

Developing efficient controlled release system of insecticide can facilitate the better use of insecticide. We described here a first example of photo-controlled release of an insecticide by linking fipronil with photoresponsive coumarin covalently. The generated coumarin-fipronil (CF) precursor could undergo cleavage to release free fipronil in the presence of blue light (420 nm) or sunlight. Photophysical studies of CF showed that it exhibited strong fluorescence properties. The CF had no obvious activity against mosquito larvae under dark, but it can be activated by light inside the mosquito larvae. The released Fip from CF by blue light irradiation in vitro retained its activity to armyworm (Mythimna separate) with LC₅₀ value of 24.64 μmol L^?1. This photocaged molecule provided an alternative delivery method for fipronil.     

Toxicity of fipronil to German and American cockroaches

Topical and oral toxicity of fipronil, compared to chlorpyrifos, was determined for the German cockroach, Blattella germanica (L.), and American cockroach, Periplaneta americana (L.). Fipronil and Combat bait matrices were evaluated for their attractancies to both species. In the topical toxicity tests, LD50's of fipronil, at 72 h after topical application, were 0.03 and 0.02 µg/g for adult B. germanica and P. americana, respectively. Fipronil was significantly more toxic than topically applied chlorpyrifos (LD50's were 0.06 and 0.16 µg/g for B. germanica and P. americana, respectively). The oral toxicity of fipronil and chlorpyrifos in Petri dish experiments, against both species, was affected by stage (for B. germanica), diet concentration, and feeding assay. Fipronil caused higher mortality of B. germanica than chlorpyrifos in two feeding assays (continuous and abbreviated). Both compounds were equally toxic to adult males of P. americana at all rates. Fipronil caused higher nymphal mortality than chlorpyrifos 48–72 h after exposure in both feeding assays. In large population chamber tests, fipronil bait was more effective and faster in killing P. americana than Raid and Combat. LT50's were 0.8, 2.4, and 7.6 d for fipronil, Raid (a.i. = chlorpyrifos), and Combat (a.i. = hydramethylnon) baits, respectively. Mortality reached 96.5, 93.4, and 84.6%, respectively, at the end of the 14 d test. In the bait attractancy tests, both strains of B. germanica were attracted similarly to fipronil and Combat bait matrices. P. americana were attracted more to fipronil than to Combat bait matrix or to other alternative foods.     

Characterization and comparative pharmacological studies of a functional γ-aminobutyric acid (GABA) receptor cloned from the tobacco budworm,Heliothis virescens (Noctuidae:Lepidoptera)

This paper reports the functional expression and pharmacological characterization of a full length complementary deoxyribonucleic acid (cDNA) (pIVY12) cloned from aHeliothis virescens fertilized egg cDNA library that encodes for a γ-aminobutyric acid (GABA) receptor subunit (HVRDL-Ser 285). Two electrode voltage clamp recordings ofXenopus oocytes expressing the HVRDL GABA-gated chloride channel revealed robust chloride ion conductance in response to GABA and the GABA_A receptor agonist, muscimol. Baclofen, a GABA_B agonist had no effect. Phenobarbital showed a positive dose-dependent allosteric modulatory effect, whereas the benzodiazepine, flunitrazepam, had no effect. Chloride conductance was depressed by the novel insecticide, fipronil ((±)-5-amino-1-(2,6 dichloro-α, α, α-trifluoro-p-tolyl)-4-trifluoromethyl-sulfinylpyrazole-3-carbonitrile) and the GABA_A antagonist, picrotoxinin. The HVRDL GABA receptor was insensitive to blockage by dieldrin and the GABA_A antagonist, bicuculline. The comparative actions of fipronil, picrotoxinin and dieldrin were examined on oocytes expressing theH. virescens wild-type (HVRDL-Ser 285), the site-directed mutant (HVRDL-Ala 285), theDrosophila melanogaster Rdl wild-type (DMRDL-Ala 302) and theRdl dieldrin resistant (DMRDL-Ser 302) homo-oligomeric GABA receptors. HVRDL-Ala 285 was 15-fold more sensitive to blockage by fipronil than HVRDL-Ser 285. DMRDL-Ala 302 and DMRDL-Ser-302 showed a similar level of sensitivity to blockage by fipronil. HVRDL-Ser 285 and DMRDL-Ser 302 exhibited a similar level of insensitivity to picrotoxinin. HVRDL-Ala 285 and DMRDL-Ala 302 showed a similar range of picrotoxinin sensitivity. DMRDL-Ala 302 and HVRDL-Ala 285 showed some sensitivity to blockage by dieldrin. Fipronil sensitivity was significantly altered by the serine to alanine mutation at position 285 in the M2 region of the HVRDL subunit, whereas no difference was observed between the DMRDL-Ser 302 and DMRDL-Ala 302 receptors.     

Palatability and toxicity of fipronil as a systemic insecticide in a bromadiolone rodenticide bait for rat and flea control

Vector control in plague-infested areas requires a simultaneous killing of rodents and their fleas. We investigated the efficacy of a combination of a systemic insecticide, fipronil, in a rodenticide bait formulation under laboratory conditions. Four different concentrations of fipronil (0.05%, 0.005%, 0.0005% with acetone as a solvent, and 0.05% with propylene glycol as a solvent) and two controls (solvents only) were combined with the rodenticide bait (crushed organically grown wheat with 0.005% bromadiolone). Each concentration was offered together with an untreated non-poisonous challenge bait to 10 singly caged Rattus rattus L., each with 100 rat fleas Xenopsylla cheopis Rothschild (Siphonaptera: Pulicidae) in the nest. Treated bait consumption was relatively low and an unsatisfactory rat mortality of around 50% only was obtained in all tests. The palatability of the bait, however, was not affected by the fipronil concentration. Even at the lowest fipronil concentration, average flea mortality was still above 95%, and doses of more than I mg fipronil per kg rat body weight gave a nearly complete kill of fleas. Fipronil can be highly effective as a systemic insecticide to for flea control, provided that a more attractive bait base for roof rats is used.     

Toxicity of 1,3-dichloropropene and fosthiazate to wireworms (Agriotes spp.)

Wireworms (Agriotes spp.) are sporadic but increasingly important pests of potatoes, sugar beet and cereals. Whilst effective chemical control is possible, the granular organophosphates normally require high rates of application and the seed dressings containing lindane (gamma‐HCH) have been withdrawn from use. The soil fumigant 1,3‐dichloropropene (1,3‐D as Telone II) and the granular nematicide fosthiazate (Nemathorin 10G) are currently used for the control of potato cyst nematodes. We investigated the effects of both of these chemicals on wireworms. Air‐vapour phase toxicities for 1,3‐D against wireworm were LD₅₀ 2.74 mg.litre.day and LD₉₉ 5.05 mg.litre.day. The in vitro soil phase toxicity was LD₉₉ 8.15 mg.litre.day. 1,3‐D soil phase activity against wireworm may be associated more with air‐vapour phase than a soil‐water phase activity. In glasshouse experiments 16.0 mg.litre.day of 1,3‐D gave 75% control. Fosthiazate, which is applied at approximately 2 μg g^?1 of soil for potato cyst nematode control, achieved an LC₅₀ at 3.20 μg g^?1. In both in vitro and glasshouse studies 1,3‐dichloropropene showed high toxicity to wireworm at dosages below the current commercial application rate for potato cyst nematode control and fosthiazate also showed useful efficacy. These chemicals may therefore prove to be valuable additional tools for limiting initial wireworm plant damage or reducing wireworm populations.     

Compatibility of entomopathogenic nematodes with fipronil

The survival and infectivity of infective juveniles (IJs) of three species of entomopathogenic nematodes, Steinernema carpocapsae Weiser, S. arenarium (Artyukhovsky) (Rhabditida: Steinernematidae) and Heterorhabditis bacteriophora Poinar (Rhabditida: Heterorhabditidae), were determined after exposure to different concentrations (250, 500, 1000 and 2000 ppm) of fipronil, an insecticide acting on the GABA receptors to block the chloride channel. Heterorhabditis bacteriophora was very tolerant to all concentrations of fipronil, with the highest mortality of 17% being observed at 2000 ppm of fipronil after 72 h exposure. Steinernema carpocapsae showed a similar response, with the highest mortality of 11.25% of IJs being observed after 72 h exposure to 2000 ppm of fipronil. Steinernema arenarium was, however, more sensitive to fipronil, and at 2000 ppm mortality rates of 94.6% and 100% were observed after 24 and 72 h, respectively. Fipronil had negligible effects on the infectivity of the three nematode species tested. The IJs which survive exposure to all concentrations of fipronil tested can infect and reproduce in Galleria larvae. The moderate effects on entomopathogenic nematodes of a lower fipronil concentration (250 ppm) and the field rates (12-60 ppm) of fipronil used as insecticide, suggest that direct mixing of entomopathogenic nematodes and fipronil at field rates is a viable integrated pest management option.     

Differential fipronil susceptibility and metabolism in two rice stem borers from China

The susceptibilities of larvae of two rice stem borers, namely, Chilo suppressalis (Walker) (Lepidoptera: Crambidae) and Sesamia inferens (Walker) (Lepidoptera: Nocutidae) to fipronil and its metabolites were investigated, and then the activities of microsomal O-demethylase, and glutathione transferase (GST) in two species were measured. The metabolism of fipronil in both stem borers was determined in vivo and in vitro. The LD50 value of fipronil to S. inferens was 118.5-fold higher than that of C. suppressalis. The bioassay results offipronil metabolites showed that the toxicities of sulfone and sulfide were higher than fipronil for both species, and the differential toxicity between sulfone and fipronil was remarkable. Alternatively, the activities of microsomal O-demethylase and GST of C. suppressalis were 1.35- and 2.06-fold higher than S. inferens, respectively. The in vivo and in vitro studies on metabolism of fipronil showed that all of fipronil, sulfone, and sulfide were detected and the content of sulfone was higher than sulfide in both stem borers. The residue of sulfone in C. suppressalis was significantly higher than that in S. inferens. These results suggest that the higher activity of mixed function oxidases may cause the higher capacity of C. suppressalis to produce fipronil-sulfone, which is more toxic than fipronil leading to the higher susceptibility of this species.     

Method of insecticide delivery affects horizontal transfer of fipronil in the German cockroach (Dictyoptera: Blattellidae).

Horizontal transmission of insecticide occurs when foragers contact or ingest an insecticide, return to the aggregation or nest, and translocate the insecticide to the shelter and its vicinity. Relatively more sedentary members of the population then contact or eat the translocated insecticide and die. We evaluated three different methods of delivering fipronil to adult male German cockroaches, Blattella germanica (L.), for their potential to cause such secondary mortality in various developmental stages of the cockroach. Adult males topically treated with 5 ng of fipronil (approximately LD99) caused low mortality in untreated nymphs and no mortality in untreated adults within the same aggregation. Males exposed to residual fipronil on a glass surface translocated more insecticide, resulting in higher mortality of cockroaches they contacted, but only early instars were affected and no adult mortality was observed. Ingested fipronil bait, however, was most effectively translocated, and caused high mortality of untreated adults and nymphs. Ingestion of fipronil also caused greater secondary kill compared with a topical application of 25 ng, approximately the same amount recovered from the exterior of males that ingested 1 mg of 0.05% fipronil bait. Secondary mortality in the untreated population was significantly affected by the duration of contact between the treated and untreated cockroaches, the quantity and freshness of excretions from the treated insects, and the accessibility of the secretions to untreated cockroaches. The mechanisms that cause secondary kill may include ingestion of excreted fipronil residues, cannibalism of bait-fed cockroaches, as well as contact with fipronil-contaminated substrates.     

Comparative toxicity,growth inhibitory and biochemical effects of terpenes and phenylpropenes on Spodoptera littoralis (Boisd.)

Eleven monoterpenes, phenylpropenes and sesquiterpenes were evaluated for their insecticidal and growth inhibitory activities against the second and fourth larval instars of Spodoptera littoralis. Among the tested compounds, 1,8-cineole revealed the highest fumigant toxicity against the 2nd and 4th larval instars with LC₅₀ values of 2.32 and 3.13 mg/L air, respectively. The monoterpenes, p-cymene, α-terpinene, (−)α-pinene and (−)-carvone were highly toxic to both larval stages as their LC₅₀ values ranged between 7.35 and 13.79 mg/L air against 2nd larval instar and between 14.66 and 32.02 mg/L air against 4th larval instar. In topical application assay against the 4th larval instar, (−)-carvone (LD₅₀ = 0.15 mg/larva) and cuminaldehyde (LD₅₀ = 0.27 mg/larva) were the most potent contact toxicants. In residual film assay, trans-cinnamaldehyde, (−)-citronellal and p-cymene showed the highest insecticidal activity against the 2nd larval instar, while α-terpinene and (−)-carvone were most effective compounds against the 4th larval instar. Moreover, the tested compounds caused strong growth reduction of both larval stages with growth inhibition higher than 80% in the 2nd larval instar and higher than 70% in the 4th larval instar. On the other hand, (−)-carvone, cuminaldehyde and (Z,E)-nerolidol showed pronounced inhibitory effects on acetylcholinesterase (AChE) and adenosine triphosphatases (ATPases) activity of S. littoralis larvae. Cuminaldehyde (IC₅₀ = 1.04 mM) and (Z,E)-nerolidol (IC₅₀ = 0.02 mM) caused the highest inhibition of AChE and ATPases, respectively. Taken together, the results indicate that monoterpenes, phenylpropenes and phenylpropenes could be used to develop new botanical insecticides for S. littoralis management.