Pharmacokinetic behavior of enantiomeric pyrethroid esters in the cockroach,Periplaneta americana L

The potent pyrethroid insecticide NRDC 157 (I; 3-phenoxybenzyl (1R, cis)-3 - (2,2-dibromovinyl)-2,2-dimethylcyclopropanecarboxylate) and its insecticidally inactive 1S, cis enantiomer (II) have similar cuticular penetration rates and are detectable in the hemolymph, nerve cord, fat body, and midgut of the American cockroach, Periplaneta americana, within 2 hr of topical application at 0.17 μg/g, a just-lethal dose of I. At this dose, both enantiomers show similar distribution patterns in these tissues, but symptoms of intoxication are seen only with I. Steady state concentrations of both enantiomers in the hemolymph and nerve cord are between 1.2 × 10^?7 and 1.7 × 10^?7M. I and II are not detectably hydrolyzed by fat body, nerve cord, and hemolymph preparations and are not detectably oxidized by fat body preparations in vitro, but a very low oxidation rate is measured for II, but not I, in nerve cord preparations. These results do not demonstrate a pharmacokinetic basis for the difference in insecticidal activity between enantiomers and therefore it is likely that the site of pyrethroid action is stereospecific. The use of the inactive enantiomer II as a model to study the effects of dose on penetration and distribution in the absence of symptoms is explored.     

Action of dieldrin and trans-aldrindiol upon the ultrastructure of the sixth abdominal ganglion of Periplaneta americana in relation to their electrophysiological effects

The action of dieldrin and trans-aldrindiol on the ultrastructure of the exposed sixth abdominal ganglion of Periplaneta americana has been studied. Dieldrin (5 μM) increased the electron opacity of the cytoplasm and nucleoplasm of glial cells without inducing such a change in the fine structure of perineurial cells. Mitochondria in nerve cell bodies and neuropile of dieldrin-treated ganglia were swollen with broken cristae and devoid of normal morphological appearance. Dieldrin treatment also caused notable depletion of synaptic vesicles from presynaptic terminals in the neuropile. Depleted terminals were found to accumulate membranous residual bodies and lysosomes indicative of insecticide-induced neuronal deterioration. These ultrastructural alterations were prevented by pretreatment of ganglia with 10mm Mg²⁺, suggesting that the action of dieldrin upon the ganglion was mediated by an increased calcium influx. trans-Aldrindiol did not exert dieldrinlike action upon the ultrastructure of the cockroach ganglia. The effects of these two chemicals on the fine structure appeared to be closely related to their electrophysiological action on the ganglion. Our studies suggest that the site and mechanism of action of dieldrin and trans-aldrindiol upon the insect nervous system may be different. The results are discussed in relation to the mode of action of cyclodiene insecticides.     

Pharmacological evidence for a discrete neurotoxic action of dieldrin (HEOD) in the American cockroach,Periplaneta americana (L.)

The excitatory effects of HEOD poisoning on spontaneous activity, and synaptic function in the sixth abdominal ganglion of the American cockroach are described. These effects are shown to be antagonized by Mg²⁺, hemicholinium-3, atropine, and d-turbocurarine. HEOD poisoning did not alter the nicotine sensitivity of the ganglion. On the basis of the findings it is hypothesized that HEOD acts at presynaptic membranes of cholinergic junctions, causing excessive and spontaneous release of presynaptic stores of acetylcholine.     

Insecticidal and repellent properties of nine volatile constituents of essential oils against the American cockroach,Periplaneta americana (L.)

The toxic and repellent properties of nine major constituents of essential oils, comprising benzene derivatives and terpenes, were evaluated against Periplaneta americana (L.). Contact and fumigant toxicities to adult females and repellency to nymphs were determined. The decreasing order of knockdown activity via contact was methyl-eugenol>isosafrole=eugenol>safrole. The killing effect via contact was in the order eugenol=methyl-eugenol=isosafrole>safrole. Fumigant toxicity was only observed for safrole and isosafrole, with safrole being the more potent. Isoeugenol and the tested terpenes had neither contact nor fumigant toxic effect. The decreasing order of repellency to nymphs was safrole>isosafrole>methyl-eugenol=α-pinene> eugenol>isoeugenol. The benzene derivatives were generally more toxic and repellent to P. americana than the terpenes. The distance of the side chain double bond from the aromatic ring and the substitution of a methoxy group to these compounds appeared to be important determinants of their toxicity and repellency. © 1998 Society of Chemical Industry     

Electrophysiological effects of paraoxon on the giant interneurons in the American cockroach,Periplaneta americana

Effects of the organophosphate insecticide, paraoxon, on the ventral grant interneurons of the American cockroach (Periplaneta americana) were studied by using intracellular microelectrodes. Within 5 min after paraoxon treatment, the efficacy of transmission at the cercal-to-giant interneuron synapse was markedly reduced or blocked, and alternating waves of depolarization and repolarization were initiated in the giant interneurons. Superimposed on these waves were high-frequency bursts of overshooting and nonovershooting spikes. Comparisons of paraoxon-induced activity in the cell body and at sites along the axon of the giant interneurons indicated that the cyclical waves, as well as high-frequency spikes, originated in the sixth abdominal ganglion rather than at more anterior sites. The effects of paraoxon were not mimicked by the application of exogenous acetylcholine, the latter causing a sustained depolarization and repetitive spiking of giant interneurons but not cyclical bursting or alternations in membrane potential. Although the precise origin of the paraoxon-induced activity is not known, several lines of evidence (e.g., results from chronic deafferentation experiments) suggest that cholinergic inputs from cercal afferent fibers are not involved in initiating the bursting.     

Inhibitors of juvenile hormone biosynthesis in corpora allata of the cockroach Periplaneta americana (L.) in vitro

The spontaneous biosynthesis of methyl (2E,6E)-(10R)-10,11-epoxy-3,7,11-trimethyldodeca-2,6-dienoate (juvenile hormone III; JH III) in excised corpora allata of Periplaneta americana was inhibited by a number of synthetic prop-2-ynyl ethers and 1,3-benzodioxole derivatives. One structurally diverse group of compounds inhibited only the final biosynthetic enzyme methyl farnesoate epoxidase (EC.1.14.14.–) at low to moderate concentrations, but at higher concentrations, also inhibited methyl farnesoate (MF) formation, causing an accumulation of MF in the concentration range 10 to 100 μM. For a second, more limited, group of compounds, there was close congruence between the inhibition of JH III biosynthesis and that of total ester (MF plus JH III) biosynthesis over their effective inhibitory concentration ranges. In contrast to the first group, there was no accumulation of MF and these compounds evidently inhibited JH III biosynthesis, at the level of either farnesoic acid esterification by O-methyl transferase (EC.2.1.1.–), or at an earlier step in the biosynthetic pathway that remains to be elucidated.     

Cholinergic properties of pinched-off synaptic nerve endings from the central nervous system of the cockroach Periplaneta americana

The preparation and cholinergic properties of a subcellular fraction, enriched in pinched-off nerve-endings (synaptosomes) from the central nervous system of the cockroach Periplaneta americana, are described. The endings retained their cytoplasmic components, as shown by the presence of marker enzymes and by ultrastructural examination. A carrier-mediated, high-affinity uptake of [³H] choline into the synaptosomes was demonstrated, and this uptake was saturable, depended on a Na⁺-gradient, and was inhibited by hemicholinium-3. It had an apparent K_m value of 0.6 (±0.1) μM, and a V_max of 20.5 (±2.5) pmol min^?1 per mg of protein. The high-affinity [³H]choline uptake was associated with the synthesis of [³H]phosphocholine and [³H]O-acetylcholine. The rate of [³H]choline uptake in synaptosomes was increased by DDT [1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane] at 100 nM concentration, and this increase was inhibited by tetrodotoxin, while neostigmine appeared to be a potent inhibitor (I₅₀ = 10 pM) of the DDT-activated uptake of [³H]choline. The site of action of the insecticides was specifically on the pre-synaptic nerve terminals because the synaptosomes preparation did not retain the post-synaptic membrane of the original nerve-endings. Cockroach synaptosomes provided a useful in-vitro preparation for studying the effects of insecticides on the pre-synaptic nerve endings.     

Toxicity and electrophysiological effects of Spilanthes amella Murr. extracts on Periplaneta americana L.

The potential insecticidal activity of Spilanthes acmella Murr. (Compositae) extracts was assessed in this study. The extracts were shown to be toxic against adults of American cockroach, Periplaneta americana L. The active component in S. acmella extract was isolated and identified as N-isobutyl-2,6,8-decatrienamide (spilanthol). The results from topical application of spilanthol show that it has a high acute toxicity and spilanthol was the most potent compound when compared to three conventional insecticides. The potency was found to be 1.3, 2.6 and 3.8 times more toxic than carbaryl, bioresmethrin and lindane, respectively. Electrophysiological studies indicated immediate hyperexcitation followed by complete inhibition of the cockroach cercal nerve activity.     

Effects of topical and bath applications of the insecticide deltamethrin on electrical activity in a leg mechanoreceptor of the cockroach,Periplaneta americana

Two different methods were used to analyze the effects of topical and bath applications of low concentrations of deltamethrin. The first method enabled the analysis of the receptor potential as well as that of the spike activity generated by mechanical or electrical stimulation of the mechano-receptor. The second method enabled recording of the conducted spike activity originating from a mechanical stimulation of the receptor and propagated along the sensory nerve. Topical application of small amounts of deltamethrin (10⁻⁹ to 10⁻⁷ g) had little effect on the receptor potential induced by mechanical stimulation of the sensory hair but blocked the action potentials within a few minutes. Electrical stimulation of the receptor cell revealed that conduction in the dendrite was affected first by the insecticide. The effects of topical application on conducted activity were compared with those of bath applications (2 × 10⁻⁸ to 2 × 10⁻⁷ M) and no significant difference was found, suggesting a rapid penetration of the insecticide through the cuticle. These effects were not reversible and this absence of reversibility was not correlated with the integrity of the barrier which protects the receptor cell from rapid changes in the ionic composition of the hemolymph. Deltamethrin was never found to induce bursts of activity in the mechanoreceptor cell under investigation either at rest or following mechanical or electrical stimulation. There are, however, some indications that other receptor cells may respond differently to this insecticide.     

Microsomal oxidase activities in relation to age and chlorcyclizine induction in American cockroach,Periplaneta americana,fat body,midgut and hindgut

Female adult American cockroaches, Periplaneta americana L., showed definite age-dependent changes in levels of activity of the microsomal mixed-function oxidases. Cytochrome P-450 levels, EPN-detoxication, and p-nitroanisole O-demethylation activities were very low in young adult insects but increased steadily reaching a natural peak at about 100 days in fat body and at about 90 days in midgut and hindgut. The activities then declined rapidly reaching levels of young insects at about 130 to 140 days of age. NADPH-neotetrazolium-reductase activity was high in young insects, declined later in adult life, and returned to a peak at about 100 days.Injections of chlorcyclizine, a known microsomal enzyme inducer, significantly increased levels of cytochrome P-450, EPN-detoxication, p-nitroanisole O-demethylation, and NADPH-NT-reductase activities in young cockroaches. The drug injections were effective, however, only before the natural activity peak was reached. Beyond this point the injections had no inductive effect indicating that the microsomal oxidases in this insect are uninducible when normal enzyme levels are falling.NADPH-NT-reductase activity in male cockroaches, while being somewhat higher than in females, showed a similar age-dependent curve with the peak occurring at about 120 days.Age-dependent carbaryl resistance in male and female insects tended to follow levels of the microsomal oxidase activities. Fifty to 60-day-old insects, however, tended to be more resistant to the insecticide than microsomal enzyme levels would indicate.RNA levels of normal female insects showed age-dependent curves similar to those of the microsomal enzyme activities, being low in young adults and reaching a peak at about 100 days. Chlorcyclizine injections had little or no effect on total microsomal RNA levels.     

Effects of the pyrethroids bioallethrin,deltamethrin and RU-15525 on the electrical activity of a cuticular mechanoreceptor of the cockroach Periplaneta americana

A study has been made of the effects of bioallethrin, RU-15525 [5-benzyl-3-furylmethyl (1R)-cis-2,2-dimethyl-3-(tetrahydro-2-oxo-3-thienylidenemethyl)-cyclopropanecarboxylate, ‘Kadethrin’], and deltamethrin on the electrical activity, measured in vivo, of a cuticular mechanoreceptor of Periplaneta americana. The modifications induced by these pyrethroids on the membrane excitability can be classified into two groups: Type I effects (bioallethrin) are characterised by a substantial increase in the number of action potentials triggered at the initiation site by a given mechanical stimulation, by an electrical activity persisting after mechanical stimulus has been stopped (repetitive activity), and possibly, by an inhibition of excitability of the cell membrane. Type II effects (RU-15525 and deltamethrin), are characterised by an inhibition of the excitability of the initiation site. In the case of RU-15525, there was a transient spontaneous electrical activity. Both types of effects have been linked to an action on the sodium channel, particularly at the initiation site. The preparation studied, which possessed no synapses, was shown to be more sensitive to deltamethrin (which is also the most insecticidal of the three pyrethroids) than to either allethrin or RU-15525. These results suggest that it is unnecessary to envisage a main target (sodium channel) that is different for the two types of pyrethroid.     

The neuroexcitatory,convulsive and lethal effects of lindane analogs on Periplaneta americana (L.)

Lindane analogs, in which one or two chlorine atoms are replaced by various substituents, exhibit convulsive and lethal effects on Periplaneta americana (L.). Some analogs are as active as lindane. These effects are well related to the neuroexcitatory action which produces after-discharges in the central nervous system. Carbamate insecticides have potent convulsive and lethal effects on P. americana, which are also related to after-discharges produced in nerve cords. Convulsion caused by lindane analogs and carbamates is solely attributable to their neuroexcitatory effects on the nervous system, in spite of the difference in their ultimate modes of action.     

美洲大蠊和东亚飞蝗的耐缺氧力及其比较

研究并比较了美洲大蠊和东亚飞蝗在接近真空和充满CO2的环境下的耐缺氧力。结果表明：美洲大蠊和东亚飞蝗在0.06pa的真空泵中都能存活,在2～10min后其生理活动会不断减弱,直到假死或静止不动,但不会死亡,90～120min后两者的生理活动逐渐恢复,但都比正常状态下弱,生理活动都呈现反抛物线的规律,且在同一时间东亚飞蝗比美洲大蠊的生理活动要弱;美洲大蠊在充满CO2的密闭瓶中存活的时间比东亚飞蝗要长,将不同程度缺氧处理后的试虫置于空气中后,美洲大蠊恢复正常生理活动的能力和恢复后的活动强度明显比东亚飞蝗强。说明美洲大蠊和东亚飞蝗都有较强的耐缺氧力,但美洲大蠊的耐缺氧力更强。     

Interaction of formamidines with octopamine-, dopamine-and 5-hydroxytryptamine-sensitive adenylate cyclase in the nerve cord of Periplaneta americana

The interaction of chlordimeform, N¹-demethylchlordimeform [N²-(4-chloro-o-tolyl)-N¹-methylformamidine], BTS-27271 [N¹-methyl-N²-(2,4-xylyl)formamidine, the 2,4-xylyl analogue of N¹-demethylformamidine], and amitraz with octopamine-, dopamine-, and 5-hydroxytryptamine-mediated enhancement of cyclic-AMP production was investigated in nerve cord homogenates of the American cockroach Periplaneta americana. N¹-Demethylchlordimeform, BTS-27271 and amitraz simulated the actions of the monoamines in enhancing cyclic-AMP production, whereas chlordimeform showed no such agonism. Additivity studies indicated that these formamidine derivatives were interacting with dopamine- and 5-hydroxytryptamine-sensitive sites, in addition to their known interaction with octopamine-sensitive adenylate cyclase. N¹-Demethylchlordimeform (Ki= 0.2μM) and amitraz (Ki=0.5μM) inhibited the dopamine-mediated response, but only N¹-demethylchlordimeform (Ki=2μM) effected appreciable inhibition of the octopamine-response. Pharmacological characterisation of the formamidineinduced elevation of cyclic-AMP production indicated that the agonistic effects of formamidines are expressed primarily through the octopamine-sensitive adenylate cyclase, rather than the dopamine- or 5-hydroxytryptamine-sensitive sites. The results are discussed in the light of the proposal that the behavioural changes, associated with formamidine poisoning, reflect the cumulative effects of octopamine-agonism, and antagonism of the dopamine- and 5-hydroxytryptaminemediated processes.     

Effect of abamectin on electrophysiological activity in the ventral nerve cord of Periplaneta americana L.: Correlation with symptoms of poisoning and levels of toxicant in vivo

Twenty-four hours after injection of adult male Periplaneta americana with 36 ng [5-³H]abamectin, when symptoms of poisoning were apparent in most individuals, similar levels of radioactivity were found in tissue samples from the ventral nerve cord, metathoracic muscle, fat body and testis of randomly-selected insects. When insects injected with [5-³H]abamectin (36 ng) were separated after 24 h into three groups showing varying symptoms, the level of radioactivity in the ventral nerve cord was found to be significantly greater in partially-paralysed and paralysed groups (4- and 7-fold respectively) than in a treated but non- paralysed group. The mean levels of abamectin in the nerve cords of unaffected, ‘partially-paralysed’ and ‘paralysed’ insects were estimated to be 4, 23 and 37 nM respectively. Extracellular studies of the in-vitro action of abamectin (10 nM and 1 μM) on the spontaneous activity of a picrotoxin-sensitive, partially desheathed, ventral nerve cord preparation of adult male P. americana found the mean response time to be relatively slow (77 and 38 min respectively). In sheathed nerve cords, the mean response time to abamectin (1 μM) was 117 min. In all cases, treatment with abamectin increased the stimulus voltage required to evoke a response. Spontaneous activity was found to be progressively reduced in ventral nerve cord preparations from ‘partially-paralysed’ and ‘paralysed’ insects compared with ‘non-paralysed’ and untreated insects following dissection 24 h after injection of P. americana with unlabelled or labelled abamectin (36 ng), while the stimulus required to evoke activity in the ventral nerve cord showed the reverse trend. These differences in electrophysiological activity could be correlated directly with varying levels of abamectin in the nerve cords of ‘paralysed’ and ‘partially-paralysed’ insects.     

美洲大蠊Na~ -K~ -ATPase作为筛选靶标的初步研究

以美洲大蠊 (Periplanata americana)雄性成虫的中枢神经系统为试材 ,对 Na -K - ATPase作为农药筛选靶标进行了初步研究 ,表明 Na - K - ATPase活力测定的最适反应条件是 :酶源蛋白浓度 8.5μg/ m L,温度 30℃ ,p H7.4 ,反应时间 15min。同时以最佳反应系统初步研究了几种拟除虫菊酯 (南开菊酯、氯氰菊酯、溴灭菊酯、氰戊菊酯、二氯苯醚菊酯、胺菊酯 )对 Na - K - ATPase离体活性的影响 ,结果显示出不同程度的抑制作用。并对其用作农药筛选靶标的可能性进行了探讨。     

The toxicity of mptp (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) to Periplaneta americana l. and its effects on dopamine in the cerebral ganglia

MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) and MPP⁺ (1-methyl-4-phenylpyridinium ion) are potent dopaminergic neurotoxins in mammals. The mammalian toxicity of MPTP depends on its conversion, by monoamine oxidase, to MPP⁺. MPTP is toxic to cockroaches (LD₅₀ 720 μg gm^?1) and the results suggest that MPTP toxicity depends on monoamine oxidase activity at a site outside the nervous system. MPTP depletes dopamine from cockroach cerebral ganglia and MPP⁺ inhibits cockroach mitochondrial respiration. While the biochemistry of MPTP toxicity appears to be the same as in mammals it seems that insects are unable to detoxify MPTP before its action has fatal consequences outside the nervous system.     

Pyrethroid resistance and cross‐resistance in the German cockroach,Blattella germanica (L)

A German cockroach (Blatella germanica (L)) strain, Apyr‐R, was collected from Opelika, Alabama after control failures with pyrethroid insecticides. Levels of resistance to permethrin and deltamethrin in Apyr‐R (97‐ and 480‐fold, respectively, compared with a susceptible strain, ACY) were partially or mostly suppressed by piperonyl butoxide (PBO) and S,S,S,‐tributylphosphorotrithioate (DEF), suggesting that P450 monooxygenases and hydrolases are involved in resistance to these two pyrethroids in Apyr‐R. However, incomplete suppression of pyrethroid resistance with PBO and DEF implies that one or more additional mechanisms are involved in resistance. Injection, compared with topical application, resulted in 43‐ and 48‐fold increases in toxicity of permethrin in ACY and Apyr‐R, respectively. Similarly, injection increased the toxicity of deltamethrin 27‐fold in ACY and 28‐fold in Apyr‐R. These data indicate that cuticular penetration is one of the obstacles for the effectiveness of pyrethroids against German cockroaches. However, injection did not change the levels of resistance to either permethrin or deltamethrin, suggesting that a decrease in the rate of cuticular penetration may not play an important role in pyrethroid resistance in Apyr‐R. Apyr‐R showed cross‐resistance to imidacloprid, with a resistance ratio of 10. PBO treatment resulted in no significant change in the toxicity of imidacloprid, implying that P450 monooxygenase‐mediated detoxication is not the mechanism responsible for cross‐resistance. Apyr‐R showed no cross‐resistance to spinosad, although spinosad had relatively low toxicity to German cockroaches compared with other insecticides tested in this study. This result further confirmed that the mode of action of spinosad to insects is unique. Fipronil, a relatively new insecticide, was highly toxic to German cockroaches, and the multi‐resistance mechanisms in Apyr‐R did not confer significant cross‐resistance to this compound. Thus, we propose that fipronil could be a valuable tool in integrated resistance management of German cockroaches. © 2001 Society of Chemical Industry     

Effects of chlordimeform and lindane on monoamine levels in the central nervous system of the american cockroach, Periplaneta americana L

The effects of chlordimeform and lindane on levels of 5-hydroxytryptamine, tryptophan, and N-acetyl dopamine were studied in the cerebral ganglia of the american cockroach, Periplaneta americana. The effects of chlordimeform on nerve cord levels of 5-hydroxytryptamine, tryptophan, dopamine, and octopamine, and the effect of lindane on cerebral ganglia levels of dopamine were also investigated in this species. Topical applications of chlordimeform deplete 5-hydroxytryptamine and tryptophan from the cerebral ganglia whereas levels of n-acetyl dopamine are elevated. The effect of chlordimeform on these compounds is dose-dependent. Similar chlordimeform-induced effects are observed in the nerve cord, and octopamine levels are also depleted in this tissue following treatment with chlordimeform. A biphasic response to chlordimeform is observed in the nerve cord for dopamine levels with a 40% decrease evident after 2 hr and a 30% increase apparent after 6 hr. In contrast to chlordimeform, lindane does not affect 5-hydroxytryptamine and tryptophan levels in the cebral ganglion but low doses of this insecticide effect increases in brain levels of dopamine and n-acetyl dopamine.     

Effect of lindane (γ-hexachlorocyclohexane) on carbohydrate and lipid reserves in the American cockroach, Periplaneta americana L

The effects of lindane on carbohydrate and lipid reserves of Periplaneta americana were studied in adult male insects. Topical application of lindane resulted in depleted levels of glycogen in the fat body (90% depletion) and thoracic musculature (57% depletion) and a 55% decrease in hemolymph trehalose (anthrone-positive material) by the prostration stage of poisoning. By contrast, lindane caused a 42% elevation of fat-body acylglycerol reserves and an associated 60% decrease of hemolymph free fatty acid levels. The lindane effects on carbohydrate and lipid were expressed also in head-ligated insects, thereby indicating that the results are not attributable solely to the action of lindane on the corpus cardiacum. The results are discussed in light of the proposal that lindane, and some other insecticides, cause indiscriminate release of neuroactive factors from the neuroendocrine system and that the consequent perturbation of physiological balance may contribute to the lethal action of the insecticide.