STUDIES OF THE TOXICITY TO WORKER HONEY-BEES (APIS MELLIFERA L.) OF CERTAIN CHEMICALS USED IN PLANT PROTECTION

Laboratory techniques are described for the estimation of the stomach poison, direct and residual film contact poison and fumigant poison effects of chemicals to adult worker honey-bees.
The toxicity of eleven chemicals used in plant protection has been investigated by these methods. The order of effectiveness as stomach and contact poisons was: parathion, TEPP, γ-BHC, dieldrin, aldrin, chlordane, o , o -diethyl- o -ethylmercaptoethyl thiophosphate (constituent of Systox), bisdimethylamino fluorophosphine oxide, toxaphene and the sodium salts of 2:4-D and MCPA: as residual films, dieldrin, aldrin, γ-BHC, parathion, chlordane, and o , o -diethyl- o -ethylmercapto-ethyl thiophosphate (constituent of Systox); toxaphene, TEPP and bisdimethylamino fluorophosphine oxide had no measurable effect; as fumigants, dieldrin, γ-BHC, aldrin, parathion, and chlordane; the remainder had no measurable effect.     

STUDIES ON THE MECHANISM OF INSECTICIDAL ACTION OF ORGANO-PHOSPHORUS COMPOUNDS WITH PARTICULAR REFERENCE TO THEIR ANTIESTERASE ACTIVITY

The preparation of an extract of the mealworm larvae, Tenebrio molitor L. which hydrolyses ethyl butyrate and o -nitrophenyl acetate, but not acetylcholine, is described. The inhibition of this esterase by TEPP-containing materials and parathion was determined.
An enzyme that hydrolysed o -nitrophenyl acetate and was inhibited by a TEPP-containing material was demonstrated in the five other insect species used.
The relative toxicities as contact insecticides to adult Tribolium castaneum Hbst. of ten samples of TEPP-containing materials were compared with their relative activities as esterase inhibitors. There was not an exact quantitative correlation between TEPP content estimated chemically, insecticidal activity and anti-esterase activity; but the correlation was sufficient to suggest interdependence of these factors.
Eggs of Diataraxia oleracea L. and Ephestia kühniella Zell, were shown to contain an enzyme that hydrolysed o -nitrophenyl acetate and was inhibited by the TEPP-containing materials. This enzyme was present in eggs less than 24 hr. old, i.e. before there was any visible signs of development. The TEPP was shown to be toxic to these eggs and in high concentrations kills at an early stage of development before differentiation of the nervous system. This, in conjunction with the other evidence, suggests that esterases other than the choline-esterase of the nervous system are important when considering the toxic action of these compounds.
Comparison of the anti-esterase activity and toxicity of parathion and TEPP-containing materials as insecticides showed that although the TEPP materials were the more potent enzyme inhibitors, parathion was the more potent contact insecticide to five species of insects. This appears to be due to the relative instability of TEPP. The study of the rates of action of the two poisons applied at different concentrations supports this view.     

THE TOXICITY OF SOME INSECTICIDES TO FIFTH-INSTAR NYMPHS OF THE DESERT LOCUST

The toxicities of the insecticides γ-BHC, diazinon, dieldrin, DNC, heptachlor and parathion to fifth-instar hoppers of the desert locust were determined. Parathion and dieldrin were the most toxic, followed in descending order by heptachlor, γ-BHC, diazinon and DNC. The resistance of the hoppers to γ-BHC and DNC changed appreciably with age within the instar.
The effects of some factors which might influence the resistance of the insects were examined and are discussed.     

THE CUMULATIVE TOXICITY OF γ-BHC AND DIAZINON APPLIED IN SMALL DOSES TO LOCUSTS

The toxicity of γ-BHC and Diazinon when applied over a period of time to adults of the desert locust, Schistocerca gregaria (Forsk.), has been determined. The insecticides were applied in oil solutions, by means of the micro-drop syringe, to the ventral surface of the abdomen.
When doses were applied in two equal portions with a 72 hr. interval, or in four equal portions at 24 hr. intervals, no significant decrease in toxicity, in comparison with a single dose, could be detected with either insecticide. It is concluded, therefore, that if similar effects occur when locusts are sprayed with these insecticides in the field, successive spraying will be fully cumulative over a period of 72 hr. Previous work using DNC has shown that doses applied over 4 days were only half as effective as equivalent single doses and that continued dosing after 4 days brought about very little increase in kill.
The sexes were similar in resistance to γ-BHC when the doses were measured in μg./g., but females were considerably more resistant to Diazinon than males.     

Persistence and distribution of insecticidal seed-dressings for control of the wheat bulb fly, Leptohylemyia coarctata

Samples of plants and soil from two experimental sites, one clay loam and one peaty loam, were analysed chemically to try to explain differing results with γ-BHC and dieldrin seed-dressings applied to control wheat bulb fly on different soil types, and to suggest reasons for patchy plant stands and occasional failure to control the pest. Seed dressed with dieldrin yielded more than untreated seed on both sites, whereas seed treated with γ-BHC yielded as much as seed treated with dieldrin on the clay site, but it yielded less than untreated seed on peat. The chemical analyses showed that the poor performance of γ-BHC on the peat site could not be explained by its failure to persist, because the soil still contained considerable quantities of γ-BHC at the time of insect attack. Possible reasons may be sorption of the poison by organic matter making it less available, and deeper sowing, permitting larvae to reach the plants without prolonged contact with the insecticide. Bayer 38156 (O-ethyl S-p-tolyl ethyl phosphonodithioate) and trichloro-nate (O-ethyl O-2,4,5-trichlorophenyl ethyl phosphonothionate) persisted in soil long enough and were sufficiently toxic to wheat bulb fly to suggest that organophosphorus compounds might provide alternatives to chlorinated hydrocarbon insecticides for control of the fly. Analyses made on seeds dressed at the laboratory for the experiment showed that the amounts of insecticide on seeds were smaller than expected and that the amounts on individual seeds differed greatly. Of ten samples of seeds dressed commercially with y-BHC, three carried very little insecticide, and the variations in the other seven samples were greater than with experimentally applied dressings.     

THE CONTROL OF POTATO VIRUS DISEASES BY INSECTICIDES

Replicated trials were carried out in the years 1950, 1951, 1953 and 1954 to see if the spread of leaf-roll virus and virus Y in potato crops could be reduced by applying insecticides. The crops were disease-free, except for small numbers of deliberately introduced infected plants, and were sprayed at intervals of 10 or 14 days, according to the stage of growth of the plants, with a tractor-mounted spraying machine at 1OO gal./acre/application. Disease spread was estimated by growing tubers taken from the five plants on either side of each infector. DDT emulsion, DDT suspension, endrin, schradan, mipafox, malathion, parathion and Systox prevented the spread of leaf-roll virus and decreased the spread of virus Y, although DDT emulsion was the only insecticide used in all the trials. Dieldrin and toxaphene were ineffective. When virus control was successful, aphid control (estimated by counting apterae in the crops) was good.     

THE USE OF SEED DRESSINGS CONTAINING γ-BHC IN THE ESTABLISHMENT OF SUGAR-BEET SEEDLINGS

From 1949 to 1952 field trials were made to determine the effect of seed dressings containing γ-BHC, with and without the addition of organic mercurial compounds, upon the establishment of sugar-beet seedlings. Most of the trials were made in fields where no serious pest problems were anticipated. The 1949 trials established the concentration of γ-BHC non-phytocidal to sugar-beet seedlings. Later trials tested the effect on pre- and post-singling plant populations of dressings applied to natural or rubbed seed sown at standardized rates. Parallel tests of dressed seed were made on farms representing a variety of soil types, It was found that the phytotoxiCity of γ-BHC varied with season and soil type, but that a dressing containing 40 % was usually safe. In these trials, the benefits from seed dressing were, as a rule, slight. Generally the combined mercury-γ-BHC seed dressing gave the best pre-singling stand of plants. Most of the improvement was attributable to the organic mercury rather than to the γ-BHC, but where pest attacks occurred, γ-BHC gave marked improvements in stand. Wireworm appeared to be the chief pest controlled: control of pygmy mangold beetle was only partial.     

ANNONA SPECIES AS INSECTICIDES

By extraction and precipitation from several solvents the toxic principle present in Annona reticulata and A. squamosa seeds and roots has been concentrated up to one hundred-fold. A preliminary chemical examination of this concentrate is described, leading to the conclusion that the toxicity is due to a glyceride or glycerides of a hydroxylated unsaturated acid or acids of high molecular weight.
These extracts have been examined for contact and stomach poison and ovicidal properties in a variety of media. When used as a contact insecticide against Aphis fabae, Macrosiphoniella sanborni and Macronphum solanifolii , the concentrate exhibited a toxicity of the same order as that of rotenone, but against Oryzaephilus surinamensis the toxicity was considerably less. As a stomach poison the ether extract was both toxic and repellent to Plutella maculipennis larvae, but was neither toxic nor repellent to Diataraxia oleracea larvae. Ovicidal tests against the eggs of Plutella maculipennis and of Ephestia kühniella were inconclusive. The potency of this concentrate is therefore of a limited nature and although of roughly the same order as that of rotenone to certain aphides, it has neither the intensity of effect nor the range of insecticidal action of that compound.     

Determination of the resistance of locusts to γ-BHC and diazinon in relation to their age, sex and weight

The relation between body weight and resistance to γ-BHC and diazinon was studied in experiments using the desert locust ( Schistocerca gregaria ) and the African migratory locust ( Locusta migratoria migratorioides) .
In all experiments with Schistocerca , resistance was proportional to weight within any one age and sex group. Resistance to diazinon (per unit body weight) decreased with age, that of the males at a faster rate than that of the females. Against γ-BHC the insects showed a nearly twofold increase in resistance about 1 week after fledging.
Within groups of Locusta the resistance to diazinon was found to be independent of weight, but the resistance to γ-BHC was dependent on weight. Similarly the resistance to diazinon at different ages after fledging was independent of age but resistance to γ-BHC increased with age.     

THE EFFECTS OF CONCENTRATION OF POISON AND THE AMOUNT APPLIED ON THE TOXICITY OF INSECTICIDAL DUSTS TO ADULT TRIBOLIUM CASTANEUM HBST

Using insecticidal dusts of D.D.T., γ-benzene hexachloride and D.N.C. under the experimental conditions, comparatively large changes in the amount of material of a given concentration are necessary in order to give appreciable differences in mortality, but small changes in the concentration of the poison will produce large differences in toxicity with a given amount applied. This relationship would appear to hold with a number of insecticides.     

Characterization of mosquito larval habitats and assessment of insecticide-resistance status of Anopheles gambiae senso lato in urban areas in southwestern Ghana

The study was carried out to characterize potential larval habitats in the city of Sekondi with the aim of assessing the relative importance of anthropogenic and natural water bodies as larval habitats. Insecticide-resistance status of Anopheles gambiae senso lato in the southwestern part of the coastal savannah zone in Ghana was also assessed against four different classes of insecticides. Larval surveys were carried out in two communities that are separated by a lagoon. Although the lagoon was a potential mosquito larval habitat, we showed that it was not an important mosquito breeding site. The major larval habitats were anthropogenic, resulting from human behavior. Some of the organically polluted breeding sites were inhabited by both An. gambiae s.l. and Culex quinquefasciatus larvae. The data also showed that An. gambiae s.l. has currently developed a strong resistance to DDT and pyrethroid insecticides in southwestern Ghana, where the species was reported to be susceptible about a decade ago. The use of insecticides in households was implicated as a possible cause of the development of resistance among An. gambiae s.l. populations in the area. The management of insecticide resistance among malaria vectors needs urgent attention if insecticide-treated materials can continue to be used for malaria control.     

Ion channels: molecular targets of neuroactive insecticides

Many of the insecticides in current use act on molecular targets in the insect nervous system. Recently, our understanding of these targets has improved as a result of the complete sequencing of an insect genome, i.e., Drosophila melanogaster. Here we examine the recent work, drawing on genetics, genomics and physiology, which has provided evidence that specific receptors and ion channels are targeted by distinct chemical classes of insect control agents. The examples discussed include, sodium channels (pyrethroids, p,p′-dichlorodiphenyl-trichloroethane (DDT), dihydropyrazoles and oxadiazines); nicotinic acetylcholine receptors (cartap, spinosad, imidacloprid and related nitromethylenes/nitroguanidines); γ-aminobutyric acid (GABA) receptors (cyclodienes, γ-BHC and fipronil) and L-glutamate receptors (avermectins). Finally, we have examined the molecular basis of resistance to these molecules, which in some cases involves mutations in the molecular target, and we also consider the future impact of molecular genetic technologies in our understanding of the actions of neuroactive insecticides.     

INSECTICIDES FOR PREVENTING BARNACLE SETTLEMENT

During the summer of 1950 experiments were made to test the use of insecticides for preventing the fouling of oyster spat collectors by the barnacle Elminius modestus.
The insecticides used were DDT, BHC (benzene hexachloride-γ-isomer) and dieldrin (hexachloro-epoxy-octahydro-dimethano-naphthalene), made up as 5% solutions and tested on egg-box collectors and quarry tiles. The tiles were used for frequent observations on settlement.
When the solutions were sprayed on to the surfaces, in sufficient quantity to give a covering of 200 mg./ft.² of active insecticide, none of them interfered with either barnacle or oyster settlement.
When the test surfaces were dipped in the solutions, DDT and BHC prevented barnacle settlement almost completely, while dieldrin reduced it by approximately 50%. Settlement of Ostrea edulis was also reduced on the treated surfaces.
Urea-formaldehyde resins made up as varnishes, to which DDT or BHC had been added, were also tested. A varnish incorporating 20% by weight of DDT provided an effective anti-fouling compound which lasted at least 3 months and completely prevented barnacle settlement.
The possibility of insecticides dissolving into the water and inhibiting barnacle growth on adjacent surfaces is discussed.     

棉铃虫的谷胱甘肽S-转移酶(GSTs):杀虫药剂和植物次生性物质的诱导与GSTs对杀虫药剂的代谢

棉铃虫Helicoverpaarmigera（Hubner）中肠谷胱甘肽S-转移酶（GSTs）对甲基对硫磷和灭多威的代谢能力明显高于对马来酸二乙酯（DEM）和两个混剂。LD5剂量的对硫磷和灭多威对棉铃虫3龄幼虫GSTs的活性均没有诱导增加的影响,用LD50的选择剂量仅对硫磷组GSTs活性增加15％。用含0．01％的芸香苷、2-十三烷酮和槲皮素的人工饲料饲养棉铃虫经1～4代后,GSTs活性提高4～18倍。3种植物次生性物质诱导组对灭多威和溴氰菊酯的敏感度均没有明显的变化,而槲皮素组对甲基对硫磷的敏感度则降低近一半,芸香苷和2-十三烷酮组对甲基对硫磷的敏感度略有降低。这种对甲基对硫磷敏感度的变化可能与上述GSTs活性的变化有关。     

A technique for determining the stomach poison effect of insecticides used against leaf-eating insects

Methods devised for feeding individual insects with leaf areas bearing known deposits of insecticides are described, and the problems associated with incomplete consumption of the treated leaf portions are discussed. An account is given of the difficulties of applying the technique to small insects and to those with erratic feeding habits.
Experimental data are given to illustrate the effects on insect resistance of the following: (i) rearing conditions, (ii) larval age and body weight, (iii) acute and chronic poisoning, (iv) the diluting effect of the leaf tissue consumed with the poison dose.
The dosage mortality curve for stomach poisons was found to be typically sigmoid: analysis of results by the method of probits (Bliss, 1934) therefore proved satisfactory. Using lead arsenate against fifth-instar larvae of Phlogophera meticulosa the ratio of weight increase to increase in median lethal dose was found to be constant.     

STUDIES IN THE MODE OF ACTION OF INSECTICIDES

Acetylcholine, choline chloride, acetyl-β-methylcholine, benzoylcholine, carbamyl choline, adrenaline and d -tubocurarine are non-toxic when injected into the locust. Prostigmine is also non-toxic, and eserine considerably less toxic to the locust than to man.
The toxic effect of tetraethyl pyrophosphate (TEPP) cannot be antagonized by injection of atropine or enhanced by d -tubocurarine.
The injection of acetylcholine chloride following injection of TEPP does not affect subsequent mortality.
These findings are discussed, and it is suggested that the physiology of the nervous system of the insect is unlike that of the mammal, neither cholinesters nor adrenaline being concerned in it. Phosphorus insecticides are thought to inhibit a general esterase not specifically connected with cholinesters.     

INSECTICIDAL CONTROL OF PAEDOGENETIC CECID LARVAE IN MUSHROOM BEDS

γ-BHC, incorporated at 50 p.p.m. in mushroom compost during the last turn of composting delayed the reproduction of paedogenetic cecid larvae without reducing the yield of mushrooms. Other insecticides were less satisfactory as they either did not control the larvae, were not sufficiently persistent or decreased the yield. Large cecid populations did not depress yields on normal composts but a proportion of the larvae migrated on to the sporophores, thus reducing the value of the crop.     

DETERMINATION OF THE RESISTANCE OF LOCUSTS TO DNC IN RELATION TO THEIR WEIGHT, AGE AND SEX

Experiments have shown that the resistance of adults of the desert locust ( Schistocerca gregaria ) and the African migratory locust ( Locusta migratoria migratorioides ) to DNC applied as a contact poison to the ventral surface of the abdomen is directly related to their body weight, irrespective of age or sex.
Varying the site of application of the insecticide had little effect on the susceptibility of the insects except when the dose was applied to the wings or femur, these being less effective sites of entry for the poison.
Increasing the post-treatment temperature from 25 to 36°C. had no noticeable effect on mortality, but the speed of action of the insecticide was much increased.     

Cost-comparison of DDT and alternative insecticides for malaria control

In anti-malaria operations the use of DDT for indoor residual spraying has declined substantially over the past 30years, but this insecticide is still considered valuable for malaria control, mainly because of its low cost relative to alternative insecticides. Despite the development of resistance to DDT in some populations of malaria vector Anopheles mosquitoes (Diptera: Culicidae), DDT remains generally effective when used for house-spraying against most species of Anopheles, due to excitorepellency as well as insecticidal effects. A 1990 cost comparison by the World Health Organization (WHO) found DDT to be considerably less expensive than other insecticides, which cost 2 to 23 times more on the basis of cost per house per 6 months of control. To determine whether such a cost advantage still prevails for DDT, this paper compares recent price quotes from manufacturers and WHO suppliers for DDT and appropriate formulations of nine other insecticides (two carbamates, two organophosphates and five pyrethroids) commonly used for residual house-spraying in malaria control programmes. Based on these 'global' price quotes, detailed calculations show that DDT is still the least expensive insecticide on a cost per house basis, although the price appears to be rising as DDT production declines. At the same time, the prices of pyrethroids are declining, making some only slightly more expensive than DDT at low application dosages. Other costs, including operations (labour), transportation and human safety may also increase the price advantages of DDT and some pyrethroids vs. organophosphates and carbamates, although possible environmental impacts from DDT remain a concern. However, a global cost comparison may not realistically reflect local costs or effective application dosages at the country level. Recent data on insecticide prices paid by the health ministries of individual countries showed that prices of particular insecticides can vary substantially in the open market. Therefore, the most cost-effective insecticide in any given country or region must be determined on a case-by-case basis. Regional coordination of procurement of public health insecticides could improve access to affordable products.     

THE INSECTICIDAL ACTIVITY OF PARATHION, ITS ISOMERS AND SOME RELATED COMPOUNDS

Heat treatment of parathion at 140° C. and above resulted in isomerization and then thermal decomposition; the loss of toxicity to Calandra granaria being correlated with a reduction of the thiono-sulphur content. Similar results were obtained with O:O-dimethyl-(4-nitrophenyl)-thiophosphate. Paraoxon, O:S-diethyl-(4-nitrophenyl)-thiophosphate, and O:O-diethyl-S-(4-nitrophenyl)-thiophosphate, although all possessing considerable contact activity, were less insecticidal than parathion; O:O-dimethyl-(4-nitrophenyl)-thiophosphate, on the other hand, was considerably more effective than parathion. O:O- bis (2-chloroethyl)-O-(4-nitrophenyl)-thiophosphate was considerably less toxic than parathion and appeared to have a different mode of action. Compounds, S-ethyl- bis -(4-nitrophenyl)-thiophosphate, O-ethyl- bis -(4-nitrophenyl)-thiophosphate, and triethyl thiophosphate were ineffective as contact insecticides.
In the series of compounds examined, replacement of the group P = S by P = O or alteration in size of the groups attached to the central phosphorus atom caused a reduction of insecticidal activity.