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Methods of testing and analyzing excito-repellency responses of malaria vectors to insecticides.

A new test system that includes an excito-repellency test box, test procedures, and statistical treatment of data is described. The method consists of enclosing 25 mosquitoes in an exposure chamber lined with insecticide-treated or untreated (control) test papers. Each chamber has a single portal for mosquitoes to escape to a receiving cage, and numbers escaping are manually recorded at 1-min intervals. The exposure chamber accommodates a screened, 2nd chamber that, when placed in the exposure chamber, prevents the mosquitoes from making physical contact with test papers. A full assay utilized one exposure chamber that permits physical contact with insecticide-treated papers, one chamber that permits physical contact with control papers, one chamber that prevents physical contact with insecticide-treated papers, and a 4th chamber that prevents contact with control papers. After insecticide exposure, test populations are held for observations on 24-h mortalities. A survival analysis approach is described for estimating mosquito escape rates and for comparing differences in mosquito escape rates, with or without physical contact with insecticide, among populations, insecticides, and doses of insecticide.

Animals

Effects of aerial spraying of insecticides on nontarget macrobenthos in a mountain stream.

The effects of aerial spraying of an insecticide were investigated in a mountain stream using a drift net. The concentration of fenitrothion (organophosphorus insecticide) in the river water increased to ca. 20 micrograms liter-1 3 hr after the spraying and decreased exponentially to half the peak value after 2 hr. A large number of aquatic insects were found drifting after the spraying. The total number of individuals which drifted in the daytime after spraying reached nine times the number found the previous night. The total number of species which drifted during the 24 hr following the spray increased to 43 from 17 on the previous day. Before the insecticide spraying, the drifting benthos were almost entirely made up of three species of Baetis (Ephemeroptera). However, many individuals of several species of Heptageniidae (Ephemeroptera), Apsilochorema sutshanum (Trichoptera), and Chironomidae (Diptera) drifted in addition to Baetis after the spray. A large number of young Baetis, which had not been seen in the natural night drift samples, were found drifting due to the insecticide. Natural night drift almost disappeared from the day following insecticide spraying. At the second insecticide spraying, conducted 20 days after the first, the number of individuals which drifted during the 24 hr following spraying decreased to only 0.85% of that in the first spraying although changes in insecticide concentration showed a similar pattern. Several species of Ephemeroptera were dominant among the fauna in the studied stream, and the causal relationships of this are discussed.

Aerosols

Cytogenetic effects of pesticides. IV. Cytogenetic effects of the insecticides Gardona and Dursban.

The cytogenetic effects of the insecticides Gardona and Dursban were investigated. The toxicity and ability of both insecticides to induce chromosome aberrations and sister-chromatid exchange in vitro was tested in a primary culture of mouse spleen cells, in order to assess the potential mutagenicity of both insecticides. The concentrations 10(-7)-10(-3) M were used for testing the toxic effects of the insecticides. Both Gardona and Dursban were toxic to spleen cell cultures and the percentage of viable cells decreased as the concentration of the insecticide was increased. It reached 76.8% and 77.8% of control after treatment with the highest concentration tested (10(-3) M) of Gardona and Dursban respectively. Gardona at 0.25, 0.50, 1.0 and 2.0 micrograms/ml, and Dursban at 0.50, 1.0, 2.0 and 4.0 micrograms/ml were tested for the induction of chromosome aberrations and sister-chromatid exchanges. All of the tested concentrations of both insecticides induced a high percentage of metaphases with chromosomal aberrations in cultured mouse spleen cells after 4-h treatment. The frequency of SCEs/cell increased with increasing concentration of the insecticides. It reached 11.92 +/- 0.14/cell and 13.40 +/- 0.20/cell after treatment with Gardona (2 micrograms/ml) and Dursban (4 micrograms/ml), respectively, compared with 8.2 +/- 0.19/cell and 7.6 +/- 0.15/cell in the solvent control. The presented results indicate that both Gardona and Dursban in the tested concentrations are mutagenic in mouse spleen cell cultures.

Animals

Bacillus thuringiensis pathogenicity islands encode regulatory circuits controlling insecticidal Cry toxin expression during vegetative growth.

Bacillus thuringiensis (Bt) produces insecticidal toxins, including Cry and Vip3 proteins, that are widely used for biological pest control. Cry proteins are classically expressed during sporulation under the control of sporulation-specific σ factors, whereas Vip3 is produced during vegetative growth, suggesting distinct regulatory pathways. Notably, many cry and vip3A genes are clustered within pathogenicity islands (PAIs), such as BtPAI-1. However, whether these PAIs also encode regulatory mechanisms coordinating toxin expression remains unclear. Here, we identify VipR, a BtPAI-1-encoded transcriptional regulator, as an activator of insecticidal gene expression during the vegetative phase in Bt strains HD-1 and CT-43. In these strains, VipR promotes the transcription of BtPAI-1 associated insecticidal genes, including vip3A and selected cry genes, resulting in premature Cry protein accumulation and increased insecticidal activity. In addition, VipR contributes to the vegetative-phase expression of the non-BtPAI-1 cry9Aa genes in strain BGSC 4AE1. Phylogenetic analysis revealed that vipR is widely distributed in one-third of Bt strains, and is strongly associated with PAIs. Futhermore, heterologous expression of vipR in BGSC 4J5 and HD-73 was sufficient to activate vegetative-phase transcription of some cry independently of sporulation-specific σ factor cascade. These results support a role for VipR in coordinating vegetative-phase expression of insecticidal genes in the Bt strains examined and suggest that BtPAI-1 can encode both insecticidal determinants and regulatory functions that influence their expression. These findings provide new insights into the regulatory architecture of Bt pathogenicity islands and may facilitate the engineering of strains with enhanced insecticidal activity.

Bacillus thuringiensis

Insecticide synergists: role, importance, and perspectives.

Synergists have been used commercially for about 50 years and have contributed significantly to improve the efficacy of insecticides, particularly when problems of resistance have arisen. In the current article we review the nature, mode of action, role in resistance management, natural occurrence, and significance in research of insecticide synergists. These natural or synthetic chemicals, which increase the lethality and effectiveness of currently available insecticides, are by themselves considered nontoxic. The mode of action of the majority of synergists is to block the metabolic systems that would otherwise break down insecticide molecules. They interfere with the detoxication of insecticides through their action on polysubstrate monooxygenases (PSMOs) and other enzyme systems. The role of synergists in resistance management is related directly to an enzyme-inhibiting action, restoring the susceptibility of insects to the chemical, which would otherwise require higher levels of the toxicant for their control. For this reason synergists are considered straightforward tools for overcoming metabolic resistance, and can also delay the manifestation of resistance. However, the full potential of these compounds may not have been realized in resistance management. Synergists have an important role to play in the ongoing investigation of insecticide toxicity and mode of action and the nature of resistance mechanism. They also can be used in understanding the effects of other xenobiotics in non-target organisms. The search for and the need of new molecules capable of synergizing existing or new pesticides has reactivated the identification and characterization of secondary plant compounds possessing such activity. Plants do possess and utilize synergists to overcome the damage produced by phytophages. This has to be exploited in pest management programs. Hopefully, it will lead to a new perspective on the nature and significance of synergism.

Animals

Continuous, alternating, and mixed insecticides affect development of resistance in the horn fly (Diptera: Muscidae).

Susceptible populations of horn flies, Haematobia irritans (L.), were selected with six insecticide treatment regimens: continuous use of permethrin, diazinon, or ivermectin; permethrin/diazinon (1:2) mixture; permethrin-diazinon rotation; and permethrin-ivermectin rotation. Insecticide selection was applied as low-volume sprays on steers infested with horn flies housed in environmentally controlled rooms. During the study period, horn fly populations developed an apparent resistance to all insecticide treatment regimens. Selection with permethrin, diazinon, and ivermectin resulted in development of resistance in generations 21, 31, and 30, respectively. Selection with insecticide mixtures and rotations resulted in the delay of any apparent resistance development for an additional one to seven generations. The magnitude of this resistance ranged from 1,470-fold for the single continuous use of permethrin to < 3-fold for ivermectin in the permethrin-ivermectin rotation. Compared with single continuous insecticide selections, the onset and degree of resistance development was significantly reduced by alternating and mixing insecticides.

Animals

Insecticide susceptibility of some vector fleas and mosquitoes in Burma.

Rat fleas and mosquitoes are insect vectors of public health importance in Burma. Plague is endemic in Central Burma and DDT has been the principal insecticide used for its control to date. Dengue haemorrhagic fever, recently introduced and transmitted by Aedes aegypti, has been spreading to major towns since 1971. The rodents, Rattus rattus, R. exulans, Bandicota bengalensis, Mus musculus, as well as shrews were commonly caught during routine trapping in the country. Rattus norvegicus, prevalent in Rangoon City, is not found in Central Burma. The rat fleas, Xenopsylla cheopis and X. astia, were found to be infesting these rodents, the former being the principal vector of plague. Insecticide susceptibility tests have been carried out periodically in plague endemic areas and Rangoon since 1966 and it is now observed that rat fleas in most of these endemic towns and Rangoon Port are no longer susceptible to DDT. Subsequent rechecks in towns where fleas became resistance to DDT show that fleas are persistently resistant to the insecticide. Aedes aegypti is found to be highly prevalent both in rural and urban areas of almost every major town and townships below 900 meters. Insecticide susceptible tests on this mosquito in limited areas show that the mosquito is generally resistant to DDT but susceptible to other insecticides. With the development of DDT resistance in fleas, it is now necessary to change to an effective insecticide in the control of plague in Burma.

Aedes

The scope and limitations of insecticide spraying in rural vector control programmes in the states of Karnataka and Tamil Nadu in India.

The resurgence of malaria in India began in 1966 and the states of Karnataka and Tamil Nadu were no exception to this phenomenon. In both states the peak occurrence came in 1976. Malaria was largely confined to highly vulnerable and receptive areas. The problem of increased incidence was particularly associated with the development of several irrigation and hydro-electric schemes. Improperly maintained irrigation systems and reservoirs provided ideal breeding grounds. The present paper examines the scope and limitations of a major anti-malaria activity, namely residual insecticide spraying as adopted and practised in rural vector control programmes in irrigation development project areas. Past experiences (as during the National Malaria Eradication programme, 1958-1965) and current practices are reviewed on the basis of selected examples. Eradication programme, 1958-1965) and current practices are reviewed on the basis of selected examples. In view of the current re-emergence of the disease, the states are faced with new obstacles to residual insecticide spraying such as (a) the development of resistance of malaria vectors to DDT and other alternative compounds like BHC (benzene hexachloride), changing vector behaviour with avoidance of contact with indoor insecticide deposits on walls, (c) environmental contamination (risks of chemicals), (d) extensive use of insecticides and pesticides for crop protection under an expanding green revolution agricultural technology, particularly in irrigated areas and (e) the existence of outdoor resting populations of the major vector Anopheles culicifacies and their role in extra-domiciliary transmission, making residual insecticide spray less effective. Spraying operations are also hindered by the persistence of certain social and cultural factors. The custom of mud plastering, white-washing and rethatching rural houses, for example, results in the loss of insecticide-treated surfaces. Other outdoor rural activities persist as obstacles in attempts to break the transmission cycle; washing, bathing and sleeping outdoors; illegal fishing and woodcutting at night; poorly constructed make-shift structures;housing project labourers near water sources; cattle grazing in nearby forests and human population movements related to seasonal migrants. The chain and extent of the transmission is dependent upon the malaria parasite carriers in the community (both indigenous and imported types) and the degree of contact of the community with those sites where people carry on the above activities, and on the effectiveness of surveillance operations.

Anopheles

Insecticide-induced changes in secretory activity of the thyroid gland in rats.

The present study was conducted to examine the effect of organochlorine (Heptachlor, Benzene hexachloride (BHC)), organophosphorus (Malathion, Monitor) and pyrethroid (Karate, Talstar) insecticides on the thyroid secretory function in rats. Heptachlor (0.5 mg per rat), BHC (0.66 mg per rat) Malathion (0.06 mg per rat), Monitor (0.2 mg per rat), Karate (0.2 mg per rat), Talstar (0.5 mg per rat) were orally administered to young adult rats for 21 days. Serum concentrations of triiodothyronine (T3), thyroxine (T4) and thyrotrophin (TSH) were determined by using specific radioimmunoassays. Body weight was not affected by treatment with any insecticide except Talstar (P < 0.01). Among organochlorine and organophosphorus insecticides, treatment with BHC and Malathion, respectively, led to a significant decrease (P < 0.01) in serum concentration of T3 and T4. Administration of BHC and Malathion also increased (P < 0.01) TSH secretion. Treatment with both of the pyrethroid insecticides similarly induced significant suppression (P < 0.01) of serum T3 and T4 levels, and concomitant stimulation (P < 0.01) of TSH concentrations. The T4/T3 ratio was decreased (P < 0.05) in rats treated with Karate but not with any other insecticide. These data indicate that immense care is warranted in the use of insecticides, because they not only affect the liver, kidney and other organs but also may alter the activity of the endocrine glands.

Animals

Susceptibility of immature Ixodes scapularis (Acari: Ixodidae) to desiccants and an insecticidal soap.

Efficacy of commercially available formulations of desiccants (Drione, Dri-die, diatomaceous earth) and insecticidal soap (Safer's) comprised of active ingredients were evaluated against Ixodes scapularis Say immatures in petri dish and grass turf bioassays. In petri dish bioassays at label rate, all products, except diatomaceous earth, provided a high degree of control by 24 and 48 h (71-100% mortality). Mortality of larvae exceeded 94% by 4 h after treatment with Drione (10 x label rate held at 85% RH), Dri-die (label rate held at both 85% and 98% RH) and Safer's insecticidal soap (10 x label rate held at 98% RH). Nymphal mortality was highest by 4 h after treatment with Safer's insecticidal soap (up to 96%). Larvae were more susceptible than nymphs to desiccants, however, nymphs were more susceptible than larvae to Safer's insecticidal soap. Immature ticks treated with Safer's insecticidal soap exhibited sublethal effects with significantly decreased attachment to hosts and no engorgement. In grass turf bioassays, mortality of nymphs was equivalent (85-95%) between treatments of chlorpyrifos (0.6 kg [AI]/ha, Safer's insecticidal soap, and Drione. In the same bioassays, nymphal mortality was moderate after treatment with Dri-die (23-29%) and low following application with diatomaceous earth (16%) and for untreated controls (6%).

Animals

Insecticides, polychlorinated biphenyls and metals in African lake ecosystems. I. Hartbeespoort Dam, Transvaal and Voëlvlei Dam, Cape Province, Republic of South Africa.

Concentrations and distribution of chlorinated hydrocarbon insecticides, polychlorinated biphenyls (PCB's) and some metals were determined in two South African lakes, Hartbeespoort Dam and Voëlvlei Dam. Water, bottom sediments, aquatic plants, aquatic insects, fish, fish-eating birds and their eggs were collected. Insecticides and PCB's were analyzed by thin layer and gas chromatography and mass-spectrometry. Analysis of metals was accomplished with atomic absorption spectrophotometry. Metals included arsenic, cadmium, copper, mangenese, lead, zinc, and mercury. The insecticide residue most commonly found in both dams were DDE, DDD, DDT, and dieldrin. Hartbeespoort had higher levesl than Voëlvlei of insecticides and PCB's in all types of samples common to both lakes. Concentrations of PCB's in all types of samples common to both lakes. Concentrations of PCB's having six or more chlorines increased with an increase in the trophic level. Concentrations of PCB's in the brains of the African birds were greater than the average total concentration of insecticides while the opposite was true for carcasses. Biological magnification of insecticides and PCB's occurred in both lakes. Hartbeespoort Dam had higher levels than Voëlvlei for all metals examined in bottom sediments and birds, except for copper in bird carcasses. Mercury levels in bird carcasses ranged from 2- to 5-fold greater than in fish while lead concentrations ranged from 2- to 10-fold greater.

Animals

Community structure and dynamics in desert ecosystems: potential implications for insecticide risk assessment.

Insecticides are increasingly being used in hot arid ecosystems. The evaluation of the ecological risk these insecticides may pose, however, is based largely on data derived from temperate organisms and ecosystems. The major differences in the composition, structure, and functioning of desert animal communities when compared to temperate terrestrial ecosystems are discussed. Desert communities are characterized by a high fraction of ectotherms (both vertebrates and invertebrates);rodents and insectivores appear to dominate the mammalian fauna; and detritivores make up a very large part of the arthropod fauna. Presently available toxicity data cover these groups only to a very limited extent. It is not known if the ranges of insecticide susceptibility observed in temperate species are representative of those in arid organisms. Thus, it is not certain that ecotoxicological assessments based on such data sets adequately protect desert animal communities. It is shown that food web connectance is higher in desert ecosystems than in temperate grasslands or forests. This may to a large extent be due to the high degree of omnivory among desert organisms. Population regulation between predators and prey appears to be weaker in deserts. The same is often, though not always, the case for competition among desert organisms. It is argued that such characteristics will reduce the chance that strong indirect effects of insecticide perturbations will occur. In spite of the fact that many desert organisms are well adapted to cope with high temporal and spatial environmental variability, there is no reason to believe that they will always recover more rapidly from population perturbations caused by insecticides. The relatively large physiological and life-history plasticity encountered in many desert animals may increase tolerance to insecticide stress. Food chains are longer in deserts than in temperate grasslands and forests. The implications of this observation for the risk of biomagnification of contaminants are discussed.

Agriculture

A malaria control trial using insecticide-treated bed nets and targeted chemoprophylaxis in a rural area of The Gambia, west Africa. 5. Design and implementation of the trial.

A large-scale malaria intervention programme using insecticide-treated bed nets and chemoprophylaxis administered to children was introduced into a rural area of The Gambia. The operation was carried out using the existing primary health care (PHC) service in the region. Training of the village health workers, sensitization of the communities, and implementation of net impregnation and the drug delivery programme are described. This delivery system resulted in over 90% of nets being treated with insecticide and 80% of children receiving over 90% of their tablets during the rainy season. There was considerable variation in the distribution of permethrin on a bed net and between individual nets, which is likely to facilitate the spread of insecticide resistance in the local mosquito populations. Bed nets made from heavier fabrics tended to absorb more insecticide than those made from lighter materials. Four months after dipping, 89% of the insecticide had been lost from treated nets. This was probably due mainly to women washing their nets, an activity carried out on average once every 2 months during the rainy season. The high number of insecticide-treated bed nets in the study area demonstrated that a malaria control programme operated through a PHC system can be implemented successfully.

Antimalarials

Biochemical mechanisms contributing to species differences in insecticidal toxicity.

Comparison of published LD50 or LC50 levels for a variety of insecticides in several vertebrate species indicate that a wide range of toxicity levels exist, and these cannot be easily predicted within either a chemical group or within a species. There is a relatively limited data base documenting interactions between insecticides and other chemicals, either agricultural or non-agricultural; however, the fact that all major insecticide groups perturb nervous system function as their primary mechanism of acute toxicity suggests the potential for interactions. Studies in our laboratories on a select group of phosphorothionate insecticides in rats indicated that brain acetylcholinesterase sensitivity to inhibition by the oxons, the active metabolites of the phosphorothionates, does not correlate with acute toxicity levels. The activities and properties of hepatic cytochrome P450-mediated activation (desulfuration) and detoxication (dearylation) of the phosphorothionates as well as of A-esterase-mediated hydrolysis of oxons contribute substantially to understanding the acute toxicity levels in rats, as does the sensitivity of the protective aliesterases to phosphorylation. However, in the channel catfish, the acetylcholinesterase sensitivity to oxon inhibition reflects the acute toxicity level of these same insecticides, and may be largely responsible for determining the acute toxicity level in this species. Thus, metabolism of insecticides appears to be far more influential in some species than others in determining the toxicity elicited.

Animals

Removal of organic pollutants from aqueous solution. V. Comparative study of the extraction, recovery and chromatographic separation of some organic insecticides using unloaded polyurethane foam columns.

The concentration of dissolved insecticides in aqueous media was determined by chromatographic separation on polyurethane foam columns. The results of preliminary screening tests on the removal of insecticides by the unloaded polyurethane foam indicated that a reasonable percentage of the insecticides was retained on the foam. Therefore attempts were made to extract these compounds from aqueous media using foam columns. Various parameters affecting the retention and separation of these compounds were studied, including temperature, flow-rate, pH, insecticide concentration, shaking time, sample volume and eluting solvent. The complete separation and quantitative recovery of these compounds from the foam with acetone in a Soxhlet extractor were achieved. The method can be used to preconcentrate insecticides in tap water and modified to determine dissolved insecticides in industrial and natural waters. Polyurethane foam has a good capacity for use when large volume samples need to be handled and is an inexpensive sorbent compared to other known solid sorbents.

Chromatography, Liquid

Social and cultural factors affecting rates of regular retreatment of mosquito nets with insecticide in Bagamoyo District, Tanzania.

Insecticide-treated mosquito nets have an impact on mortality and morbidity in young children under controlled conditions. When integrated into larger control programs, there is the danger that rates of regular retreatment of the nets with insecticide will drop, greatly limiting their effectiveness as a public health intervention. In Bagamoyo District, Tanzania, rates of retreatment dropped significantly when payment for the insecticide was introduced. A series of neighbourhood (hamlet) meetings were held in all study villages to discuss people's concerns about the insecticide and ways to increase rates of retreatment. Although changes were made in the procedure for retreatment, rates of retreatment remained lower than expected and showed marked variation within as well as between villages. We then conducted unstructured key informant interviews as well as informal discussions in a village with strong variation between different sectors of the villages in rates of retreatment. While logistical problems were most frequently cited as reasons not to bring nets for retreatment, political and social divisions within the community provided a better explanation. This is borne out by the low response to rearrangements in logistics which made retreating the nets significantly easier for households, and the higher response when changes were made in the channels of communication as well as the logistic features. It is clearly more difficult for villagers to appreciate the benefits of the insecticide than those of the nets. Great emphasis needs to be placed on the insecticide and its beneficial effects from the outset for any large-scale programme to be sustainable.

Bedding and Linens

Residues of insecticides, and fungicides in fruit produced in Ontario, Canada, 1986-1988.

Between 1986 and 1988, 306 composite samples of fruit representing eight commodities were collected from farm deliveries to the marketplace in Ontario, Canada. All samples were analysed for insecticides and fungicides. The analysis procedure included tests for organochlorine, organophosphorus, synthetic pyrethroid and methylcarbamate insecticides and dithiocarbamate, dicarboximide and organochlorine fungicides. The commodities tested included apples, blueberries, cherries, grapes, peaches, pears, raspberries and strawberries. In 14% of all fruit samples, pesticide residues were below the detection limits, which ranged between 0.005 and 0.02 mg/kg. A further 14% had total combined pesticide residues below 0.1 mg/kg. Total combined fungicide and insecticide residues ranged from 0.1 to 11 mg/kg in 72% of samples. Six different pesticides were in violation of maximum residue limits (MRL) on 11 (3.6%) of fruit samples. Captan exceeded the 5 mg/kg MRL in five samples and EBDC exceeded the 7 mg/kg MRL in two. Other violations included single fruit samples with dicofol, endosulphan, phosalone and iprodione above the MRL. Raw grapes harvested for wine contained residues of 10 pesticides and the number changed little following the crushing of the grapes; however, fermentation into wine significantly reduced residues. Six insecticides and four fungicides were present on the raw grapes and 4/105 were above the MRL. Following crushing, four insecticides and five fungicides were identified and 4/40 were above the MRL. In wine only three insecticides were identified and all were well below the MRL. Carbaryl appeared to be the most persistent, declining very little between raw grapes and wine.

Food Contamination

Effects of several insecticide formulations on oothecal drop and hatchability in German cockroaches (Dictyoptera: Blattellidae).

Formulations used for control of German cockroaches, Blattella germanica (L.), were investigated. The insecticides tested on gravid females were bendiocarb, chlorpyrifos, cyfluthrin, cypermethrin, fenvalerate, hydramethylnon, malathion, propetamphos, propoxur, and pyrethrins. Exposure to each insecticide increased the frequency of oothecal drop and reduced the percentage oothecal hatch. The percentage of oothecae that were dropped increased as a curvilinear function of insecticide concentration. An LD50 of propoxur to female German cockroaches resulted in the greatest oothecal drop (83.50%); fenvalerate caused the least drop (22.60%). LD50's of cypermethrin, propetamphos, and propoxur resulted in 29.60, 31.807, and 37.30% hatch from dropped oothecae, respectively. The smallest percentage hatch from retained oothecae was caused by LD50's of propoxur (1.5%) and cyfluthrin (7.70%). Retained oothecae from females treated with an LD50 of fenvalerate (68.70%) or pyrethrins (68.70%) had the greatest percentage hatch. Total percentage hatch (dropped and retained oothecae) declined exponentially as the insecticide concentration increased. An LD50 of cypermethrin limited total oothecal hatch to 24.50%, whereas LD50's of fenvalerate, malathion, and pyrethrins resulted in 53.60, 52.20, and 58.90% hatch, respectively. Mean time to oothecal hatch increased linearly with increasing insecticide concentration for all insecticides tested.

Animals