Genetic and biochemical mechanisms limiting fipronil toxicity in the LPR strain of house fly,Musca domestica

Fipronil is a new insecticide which exerts its toxic action by interacting with the insect GABA-gated chloride channel. Previous studies have shown that cyclodiene-resistant insects have low to moderate levels of cross-resistance to fipronil, while other resistant strains are usually susceptible. In contrast, we recently found a strain (LPR) of house fly (Musca domestica L) with 15-fold cross-resistance to fipronil that was not associated with cyclodiene resistance. Fipronil cross-resistance in LPR was inherited as an intermediately dominant, autosomal, multigenic trait. [¹⁴C]Fipronil was observed to penetrate into LPR flies more slowly than into susceptible flies. S,S,S-tributylphosphorotrithioate and diethyl maleate pretreatment did not reduce the level of fipronil cross-resistance, while piperonyl butoxide resulted in a slight decrease. These results indicate that decreased penetration and monooxygenase-mediated detoxification may be mechanisms contributing to fipronil cross-resistance in the LPR strain. © 1999 Society of Chemical Industry     

Glutamate-activated chloride channels: Unique fipronil targets present in insects but not in mammals

Selectivity to insects over mammals is one of the important characteristics for a chemical to become a useful insecticide. Fipronil was found to block cockroach GABA receptors more potently than rat GABA_A receptors. Furthermore, glutamate-activated chloride channels (GluCls), which are present in cockroaches but not in mammals, were very sensitive to the blocking action of fipronil. The IC₅₀s of fipronil block were 30 nM in cockroach GABA receptors and 1600 nM in rat GABA_A receptors. Moreover, GluCls of cockroach neurons had low IC₅₀s for fipronil. Two types of glutamate-induced chloride current were observed: desensitizing and non-desensitizing, with fipronil IC₅₀s of 800 and 10 nM, respectively. We have developed methods to separately record these two types of GluCls. The non-desensitizing and desensitizing currents were selectively inhibited by trypsin and polyvinylpyrrolidone, respectively. In conclusion, in addition to GABA receptors, GluCls play a crucial role in selectivity of fipronil to insects over mammals. GluCls form the basis for development of selective and safe insecticides.     

Voltage-dependent modulation of the neuronal nicotinic acetylcholine receptor-channel by cartap

Nicotinic acetylcholine receptor-channels (AChR) are believed to be the target site of cartap. In order to clarify the mechanisms of the cartap interaction with nicotinic AChRs, single-channel patch clamp experiments were performed using rat clonal phaeochromocytoma (PC12) cells. Cartap increased the short closures occurring during single-channel openings induced by acetylcholine (ACh). Thus, when co-applied with acetylcholine, cartap markedly shortened the open time associated with bursts. These effects were voltage-dependent, being more prominent at negative voltages. These results strongly suggest that cartap acts as an open channel blocker at the neuronal nicotinic AChR. © 1999 Society of Chemical Industry     

氟虫腈对家蚕的急性毒性与安全评价研究

氟虫腈是我国稻桑混栽地区水稻上普遍使用的杀虫剂。采用食下毒叶法、口器注入法与药 膜法测定了不同条件下氟虫腈对家蚕的急性毒性。氟虫腈对2龄、3龄、4龄、5龄家蚕的摄入LC50值 (48 h,下同)分别为4.89、6.46、9.47、11.59 mg/L;对5龄家蚕的LD50值为1.38 μg/蚕;在25、30、35 ℃ 3个温度下,氟虫腈对2龄家蚕的LC50值分别为4.81、4.59、2.05 mg/L;桑叶浸药时间为1 s、10 s、1 min、10 min、1 h时,氟虫腈对3龄家蚕的LC50值分别为9.12、6.46、4.39、3.68、2.80 mg/L;家蚕在氟虫腈药膜上爬行1、10、30、60 min后, 对4龄家蚕的接触LC50值分别为2.82、2.35、2.05、1.57 μg/cm2。研究表明,胃毒是氟虫腈对家蚕的主要作用方式,氟虫腈在桑园附近水稻上使用对家蚕有较大风险。     

氟虫腈对水稻害虫的作用特点及应用

通过室内生物测定和田间试验，明确了氟虫腈（商品名为锐劲特Regent）对三化暝的内吸杀虫活性、杀卵活性、持效期和对褐飞虱的杀虫活性以及对水稻主要害虫的田间防治效果，同时评价了其对天敌的安全性。结果表明：（1）氟虫腈由水稻根和茎叶内吸传导对三化暝的杀虫活性高，根和茎叶的内吸对初孵蚁暝的LC50值分别为29．12和13．07mg/L；（2）持效期长，50mg/L氟虫腈药后10d和15d对接三化暝初孵蚁暝的防效仍达92．1％和87．2％，而甲胺磷1000mg/L药后3d的防效即降为0；（3）对三化暝的杀卵效果较差，100mg/L的杀卵效果仅为30．3％，极显著低于500mg/L三唑磷的杀卵效果（95．26％）；（4）对褐飞虱的综合活性高，对3龄若虫的LC50值为10．52mg/L；（5）在江苏高淳、丹阳等地的田间试验表明，5％氟虫腈悬浮剂0．3、0．45、0．75L/hm^2可分别有效控制白背飞虱、三化暝枯心、稻纵卷叶暝、三化暝白穗和褐飞虱；（6）氟虫腈对以狼蛛为代表的稻田蜘蛛杀伤力大，但对暝卵内赤眼蜂幼期较安全。     

The A302S mutation in Rdl that confers resistance to cyclodienes and limited cross-resistance to fipronil is undetectable in field populations of house flies from the USA

Fipronil is a relatively new insecticide with great potential for insect control, however widespread use of cyclodiene insecticides has selected for an A302S mutation in the Rdl (GABA gated chloride channel) allele. This mutation gives resistance to cyclodienes and limited cross-resistance to fipronil. Given the concern over the possible reduction in efficacy and/or lifetime that fipronil might be used for pest control (given the extensive use of cyclodienes in the past), it is important to know the frequency of the A302S Rdl mutation in field populations. To ascertain the relative frequency of the A302S Rdl mutation in house fly populations we used three experimental approaches. First, we attempted to select for fipronil resistance by initially treating 33,100 field collected flies and selecting 14 additional generations. We were unable to produce a highly resistant strain. Second, we directly sequenced field collected flies. Third, we tested field collected house flies with a diagnostic dose of dieldrin and then genotyped the survivors. Out of the 4750 flies tested, there were no Rdl resistance alleles detected. We conclude that the resistant Rdl allele is rare in house flies in the US due to decades without cyclodiene use and a fitness disadvantage (in the absence of cyclodienes) of the 302S Rdl allele. The limited cross-resistance provided by the cyclodiene resistant Rdl allele, combined with the very low frequency of this allele in field populations, suggests that fipronil could be a promising insecticide for house fly control.     

The role of GABA and glutamate receptors in susceptibility and resistance to chloride channel blocker insecticides

Despite a point mutation in the pore-forming segment of the Rdl GABA receptor subunit that is widespread and persistent in insect populations and confers high levels of resistance to dieldrin and other polychlorocycloalkane (PCCA) insecticides, the phenylpyrazole insecticide fipronil, which binds at same site, has proven to be effective in controlling many insects, including dieldrin-resistant populations. Fipronil and its major sulfone metabolite are unique among chloride channel blocking insecticides in that they also potently block GluCls. We present here a patch clamp study of the action of fipronil sulfone on native GABA receptors and GluCl receptors from susceptible and dieldrin-resistant German cockroaches, to provide a better understanding of the effect of the Rdl mutation on the function and insecticide sensitivity of these two targets, and its role in resistance. Dieldrin blocked GABA currents with an IC₅₀ of 3 nM in wild-type cockroaches, and 383 nM in resistant insects, yielding a resistance ratio of 128. Fipronil sulfone blocked GABA currents with an IC₅₀ of 0.8 nM in susceptible insects and 12.1 nM, or 15-fold higher, in resistant insects. While both GluClD (desensitizing) and GluClN (non-desensitizing) receptors were found in German cockroach neurons, GluClN receptors were rare and could not be included in this study. GluClD receptors from resistant insects had reduced sensitivity to glutamate and a lower rate of desensitization than those from susceptible insects, but their sensitivity to block by fipronil sulfone was not significantly changed, with an IC₅₀ of 38.5 ± 2.4 nM (n = 8) in the susceptible strain and 40.3 ± 1.0 nM (n = 7) in the resistant strain. Fipronil sulfone also slowed the decay time course of GluClD currents. These results suggest that GluClD receptors contain the Rdl subunit, but their sensitivity to fipronil sulfone is not altered in resistant insects.     

Adsorption, transport and degradation of fipronil termiticide in three Hawaii soils

BACKGROUND: The behavior of the termiticide fipronil in soils was studied to assess its potential to contaminate ground and surface water. This study characterizes (1) adsorption of fipronil in three different soils, (2) transport of fipronil through leaching and runoff under simulated rainfall in these soils and (3) degradation of fipronil to fipronil sulfide and fipronil sulfone in these soils. RESULTS: The adsorption experiments showed a Freundlich isotherm for fipronil with K_oc equal to 1184 L kg^?1. In the leaching experiments, the concentration of fipronil and its metabolites in leachate and runoff decreased asymptotically with time. The concentration of fipronil in the leachate from the three soils correlated inversely with soil organic carbon content. The degradation experiment showed that the half‐life of fipronil in the soils ranged from 28 to 34 days when soil moisture content was 75% of field capacities, and that 10.7–23.5% of the degraded fipronil was transformed into the two metabolites (fipronil sulfide and fipronil sulfone). CONCLUSION: Fipronil showed large losses through leaching but small losses via runoff owing to low volumes of runoff water generated and/or negligible particle‐facilitated transport of fipronil. The half‐life values of fipronil in all three soils were similar. Copyright © 2011 Society of Chemical Industry     

氟虫腈对二化螟生长发育的影响及对解毒酶的诱导效应

研究了氟虫腈对不同虫态二化螟Chilo suppressalis的作用特点、持续毒性及对解毒酶的诱导效应。结果表明:氟虫腈对二化螟幼虫具有很强的毒杀作用,且毒杀作用随幼虫龄期的增长而逐渐减弱;对二化螟蛹的羽化具有较低的抑制作用;对卵孵化无抑制作用。亚致死剂量氟虫腈能引起幼虫持续死亡,降低存活试虫体重增长率和蛹重。用亚致死剂量氟虫腈处理4龄幼虫后,对其体内羧酸酯酶活性具有明显的诱导效应和时间效应,对谷胱甘肽S--转移酶活性的诱导效应仅在药剂处理后3h内较为显著,对多功能氧化酶活性表现出较强的诱导效应,比活力约为对照的1.37倍。     

Fipronil metabolism,oxidative sulfone formation and toxicity among organophosphate‐ and carbamate‐resistant and susceptible western corn rootworm populations

Fipronil toxicity and metabolism were studied in two insecticide‐resistant, and one susceptible western corn rootworm (Diabrotica virgifera virgifera, LeConte) populations. Toxicity was evaluated by exposure to surface residues and by topical application. Surface residue bioassays indicated no differences in fipronil susceptibility among the three populations. Topical bioassays were used to study the relative toxicity of fipronil, fipronil + the mono‐oxygenase inhibitor piperonyl butoxide, and fipronil's oxidative sulfone metabolite in two populations (one resistant with elevated mono‐oxygenase activity). Fipronil and fipronil‐sulfone exhibited similar toxicity and application of piperonyl butoxide prior to fipronil resulted in marginal effects on toxicity. Metabolism of [¹⁴C]fipronil was evaluated in vivo and in vitro in the three rootworm populations. In vivo studies indicated the dominant pathway in all populations to be formation of the oxidative sulfone metabolite. Much lower quantities of polar metabolites were also identified. In vitro studies were performed using sub‐cellular protein fractions (microsomal and cytosolic), and glutathione‐agarose purified glutathione‐S‐transferase. Oxidative sulfone formation occurred almost exclusively in in vitro microsomal reactions and was increased in the resistant populations. Highly polar metabolites were formed exclusively in in vitro cytosolic reactions. In vitro reactions performed with purified, cytosolic glutathione‐S‐transferase (MW = 27 kDa) did not result in sulfone formation, although three additional polar metabolites not initially detectable in crude cytosolic reactions were detected. Metabolism results indicate both cytochromes P450 and glutathione‐S‐transferases are important to fipronil metabolism in the western corn rootworm and that toxic sulfone formation by P450 does not affect net toxicity. © 2000 Society of Chemical Industry     

Effect of vegetation on the longevity,mobility and activity of fipronil applied at the termiticidal rate in laboratory soil columns

BACKGROUND: Termiticides are applied at concentrations much higher than those used in agricultural settings. The longevity of fipronil has not yet been examined at the rates used for termite control, nor has the compound's movement in the soil been addressed. RESULTS: Fipronil was detected in the eluates of treated soil cones, increasing initially and then decreasing to a steady concentration of about 1 µg mL^?1. In larger PVC pipe plots, fipronil in the top treated soil depth (0–7.5 cm) dissipated more rapidly (half‐life of 11–13 months) than in treated soil at the next treated depth (7.5–15 cm; half‐life of 20–29 months). The presence of vegetation had no significant effect on the mobility, longevity or movement into untreated depths. Treated soil remained toxic to termites throughout the duration of the study. Fipronil moved into the 15–22.5 cm soil depth in sufficient concentration to cause 100% mortality to eastern subterranean termites in 3 day bioassays. CONCLUSION: Fipronil remains in treated soil at levels toxic to termites for at least 30 months. Movement of the active ingredient was observed in sufficient amounts to kill termites in non‐treated soil directly below the treated soil. Published 2010 by John Wiley & Sons, Ltd.     

Insecticidal efficacy of fipronil against four stored‐product insect pests: influence of commodity,dose, exposure interval,relative humidity and temperature

BACKROUND: Fipronil is an insecticidal pyrazole that is commonly used as an insecticide in field crops, urban pesticide and veterinary medicine, but there are no reports of its evaluation against stored‐product insects. Three series of laboratory bioassays were conducted to assess fipronil as a potential grain protectant against adults of Sitophilus oryzae (L.), Tribolium confusum Jacquelin du Val, Rhyzopertha dominica (F.) and Prostephanus truncatus (Horn). Factors such as dose (0.01, 0.1, 1 and 10 mg fipronil kg^?1 grain), exposure interval (24 h, 48 h, 7 days and 14 days), temperature (20, 25 and 30 °C), relative humidity (RH; 55 and 65%) and commodity (wheat, maize, barley and paddy rice) were evaluated with regard to their impact on the insecticidal activity of fipronil. Progeny production was assessed after 64 days of exposure. RESULTS: At doses of < 1 mg fipronil kg^?1 grain, efficacy of fipronil was enhanced by an increase in temperature from 20 to 25 °C against S. oryzae or R. dominica for exposures of > 48 h and against T. confusum or P. truncatus for exposures of > 24 h. A further increase in temperature from 25 to 30 °C, although it enhanced efficacy at doses of < 1 mg fipronil kg^?1 grain against S. oryzae at all exposure intervals and against T. confusum after 7 days of exposure, it was negatively associated with efficacy against R. dominica or P. truncatus for exposures of > 24 h. By contrast, increase in RH did not have a significant impact on efficacy of fipronil. Although the performance of fipronil among the tested commodities was species dependent, this substance appeared to be less effective in paddy rice than in barley, maize and wheat. Regardless of the treated commodity, progeny production of all the tested species was almost suppressed with doses higher than 0.1 mg fipronil kg^?1 grain. CONCLUSIONS: Fipronil appeared to be a very effective alternative to the existing substances in stored‐grain protection at doses equal to or higher than 1 mg fipronil kg^?1 grain, and thus these doses require further evaluation in terms of safety for the consumer and residues on stored products. Copyright © 2010 Society of Chemical Industry     

Cross-resistance potential of fipronil in Musca domestica

The toxicity of fipronil to insecticide-susceptible houseflies and the cross-resistance potential of fipronil were determined for six insecticide-resistant laboratory housefly strains by topical application and feeding bioassay. The insecticide-resistant strains represented different levels and patterns of resistance to pyrethroids, organophosphates, carbamates and organochlorines. Five strains were almost susceptible to fipronil in feeding bioassay with resistance factors at LC50 between 0.36 and 3.0. Four of these strains were almost susceptible to topically applied fipronil (resistance factors at LD50 were 0.55, 0.83, 3.3 and 2.5, respectively), whereas one strain was 13-fold resistant to topically applied fipronil. A highly gamma-HCH-resistant strain, 17e, was 430-fold resistant to fipronil in topical application bioassay and 23-fold resistant in feeding bioassay at LD50/LC50. We also tested the toxicity of fipronil in a feeding bioassay and gamma-HCH in topical application bioassay on thirteen housefly field populations. Eleven of the field populations had resistance factors for fipronil ranging from 0.98 to 2.4 at LC50, whereas two populations were 4.0- and 4.6-fold resistant to fipronil. The resistance level to gamma-HCH at LD50 in the field populations ranged from 1.8- to 8.1-fold. The two strains showing fipronil resistance were 3.4- and 8.1-fold resistant to gamma-HCH. Fipronil and gamma-HCH toxicities were positively correlated in the field populations. Biochemical assays of esterase, glutathione S-transferase and cytochrome P450 monooxygenase indicated that the low fipronil resistance observed in laboratory and field strains could be caused by elevated detoxification or be due to a target-site resistance mechanism with cross-resistance to gamma-HCH.     

Insecticidal activity of Sikimi extract and its modulation of the GABAA receptor channel

Sikimi plant (also known as Japanese star anise), Illicium anisatium, is toxic to mammals. Extracts of Sikimi were studied for their insecticidal activity against the larvae of mosquito, Culex quinquefasciatus, and for their mechanism of action on ion channels. Crude methanol extract and its ethyl acetate-soluble fractions were insecticidally active, with EC₅₀ values of 63·0 μg ml^-1 and 43·7 μg ml^-1, respectively. The ethyl acetate-soluble fraction was perfused through the bathing solution and the current induced by a brief (10 ms) application of GABA by pressure ejection through pipette electrode was recorded by the whole-cell patch clamp technique. The extract suppressed GABA-induced currents irreversibly with an EC₅₀ value of 0·42 μg ml^-1. The time constant of current fitted to the single exponential function was shortened by the ethyl acetate-soluble fraction at concentrations ranging from 0·1 μg ml^-1 to 10 μg ml^-1 in a concentration-dependent manner. It was concluded that Sikimi extracts decreased the affinity of GABA for its binding site on the GABA receptor, thereby suppressing GABA-induced currents. © 1998 SCI     

In vivo and in vitro metabolism of fipronil by larvae of the European corn borer Ostrinia nubilalis

In vivo and in vitro metabolism of [14C]fipronil was examined in a susceptible European corn borer (Ostrinia nubilalis, Hübner) laboratory strain. [14C]Fipronil penetrated the larval integument slowly, with 71.5% of the applied radioactivity recovered from surface rinses 24 h after topical application. Despite this slow penetration, radioactivity was detected in both the excrement and internal organo-soluble fractions. Radioactivity in the internal aqueous fraction and tissue pellet accounted for less than 0.8% of total radioactivity. The in vivo studies suggest that fipronil oxidation to its sulfone metabolite is the major route of metabolic conversion. In vitro studies were performed using subcellular microsomal fractions isolated from European corn borer larval midguts. Cytochrome P450-dependent monooxygenase activity (methoxyresorufin O-demethylase) was consistently observed in midgut preparations, and formation and detection of the sulfone metabolite in the same midgut preparations was also NADPH-dependent and inhibited by piperonyl butoxide. In vitro metabolism results indicate microsomal monooxygenases are responsible for the conversion of fipronil to its sulfone form in the European corn borer.     

Argentine ant control around homes: efficacy of treatments and urban runoff

ABSTRACT

Pest management professionals in California receive more customer complaints about Argentine ants than for any other urban ant pest. Fipronil, applied as a 30 × 30 cm band around the house foundation, has become the preferred treatment used to control these ants. Unfortunately, fipronil is now showing up in urban waterways at levels that are toxic to aquatic invertebrates. Our recent studies are aimed at mitigating insecticide runoff while still controlling the ant infestations. A high priority is preventing fipronil runoff from the driveway to the street, where it can flow into drains and from there to urban waterways. In this paper, two related studies address these issues. Not treating driveways with fipronil reduced by two to three orders of magnitude its runoff when compared with earlier studies. However, not treating the driveway can reduce efficacy of treatments. Granular bifenthrin, indoxacarb, botanicals, and a thiamethoxam ant bait were tested as supplemental treatments. The gel bait showed the best result as a supplement, but only after 8 weeks. We have reduced fipronil runoff while maintaining efficacy of the ant treatments.     

Behavioral effects of acute exposure to the insecticide fipronil

The effects of fipronil (Frontline® Top Spot) were investigated in 40 days old rats utilizing open field (OF), hole-board (HB) and elevated plus-maze (EPM) apparatus. Rats (N = 15) received topical application of fipronil (70, 140 and 280 mg/kg) in the neck region and behavior was tested 3 h after administration. Animals treated with corn oil (vehicle) were used as controls. In the OF test animals treated with fipronil at 140 mg/kg showed increased rearing, whereas animals exposed to 280 mg/kg showed increased freezing, grooming, and rearing. In the HB test fipronil at 280 mg/kg increased head-dip and head-dipping behaviors. In the EPM test the only observed effect was increased number of entries in both open and closed EPM arms in animals treated with 280 mg/kg. In conclusion, dermal exposure to fipronil causes effects related to emotionality, fear, and exploratory activity; results add strength to the growing concern that pirazole insecticides can be neurotoxic to humans.     

Fipronil resistance mechanisms in the rice stem borer, Chilo suppressalis Walker

Fipronil resistance mechanisms were studied between the laboratory susceptible strain and the selective field population of rice stem borer, Chilo suppressalis Walker in the laboratory. The borer population was collected from Wenzhou county, Zhejiang province. After five generations of selection, fipronil resistance ratio was 45.3-fold compared to the susceptible strain. Synergism experiments showed that the synergistic ratios of PBO, TPP and DEF on fipronil in susceptible and resistant strains of C. suppressalis were 7.55-, 1.93- and 2.91-fold, respectively, and DEM showed no obvious synergistic action on fipronil. Activities of carboxylesterase and microsomal-O-demethylase in the resistant strain were 1.89- and 1.36-fold higher that in susceptible strain, and no significant difference of glutathione-S-transferase activity was found between the resistant and susceptible strains. The K_m and V_max experiments also demonstrated that fipronil resistance of C. suppressalis was closely relative to the enhanced activities of carboxylesterase and microsomal-O-demethylase. Moreover, cross-resistance between fipronil and other conventional insecticides and the multiple resistant properties of the original Wenzhou’s population were also discussed.     

Consumption of fipronil-treated rice seed does not affect captive blackbirds

Fipronil is an insecticide developed for use on rice seed and other crops. In a series of cage and pen trials, we evaluated the responses to dyed, fipronil-treated rice seed of three bird species likely to encounter it in the field. Individually caged red-winged blackbirds (Agelaius phoeniceus), brown-headed cowbirds (Molothrus ater), and boat-tailed grackles (Quiscalus major) displayed no evidence of adverse reaction to treated seed. Chemical analyses of hulls from treated seeds eaten by captive birds revealed that 10–20% of the fipronil originally present was removed during feeding. In group enclosures, male red-winged blackbirds ate as much fipronil-treated rice as they did dyed, untreated seed. In four-day tests within a 0·2-ha flight pen, 10-bird blackbird flocks removed 11·4% of fipronil-treated seed from a test plot compared to 12·5% of dyed, untreated seed removed from the alternate plot. When the alternate plot contained undyed rice, however, seed removal from the treated plot averaged 2·4% compared to 28·9% from the alternative plot, suggesting that the groups of test birds avoided treated seed based on its appearance. We conclude that 325 and 500 mg kg^-1 fipronil applications alone do not affect avian feeding activity. © 1998 SCI.     

Effect of fipronil seed treatments on the germination and early growth of rice

Fipronil seed treatments were evaluated to determine whether they directly influence germination and subsequent seedling growth in rice (Oryza sativa L). Continuous seed exposure to fipronil (four days) at 2 000 mg litre⁻¹ significantly impaired germination (P < 0.001). When exposure was restricted to a 1-h period 48 h after the initiation of germination, early post-germination growth was also impaired (assessment two days after exposure, P < 0.05). The proportion of seeds satisfying our criteria for normal germination fell by 2.3 and 2.6% respectively across 17 cultivars. Cultivar effects were highly significant (P < 0.001). When exposure to fipronil (2 000 mg litre⁻¹) was restricted to 2 h at initial seed wetting no significant growth impairment occurred. No significant differences (P > 0.05) were found between shoot lengths or root system dry weights of control plants and plants developing from seed exposed continuously (two days) to fipronil at rates of up to 2 000 mg litre⁻¹ during germination and harvested nine days after sowing. Treating germinated seed with fipronil for 1 h immediately prior to sowing at rates of up to 4 000 mg litre⁻¹ did not result in significant changes (P < 0.05) in plant growth parameters at either nine or 25 days after sowing. No evidence of fipronil having a direct phytostimulatory effect on rice was obtained. © 1999 Society of Chemical Industry