Synthesis and use of an isoform-specific affinity matrix in the purification of glutathione S-transferases from the housefly, Musca domestica (L.)

Glutathione may be linked to an agarose matrix which has been activated by treatment with epichlorhydrin. The resulting resin displayed group selectivity for the glutathione S-transferases of the housefly Musca domestica (L). The isoenzymes of low isoelectric point, which have little activity with substrates other than 1-chloro-2,4-dinitrobenzene, bound strongly to this matrix and were eluted with 10 mM glutathione at pH 7.4. On the other hand, the group of isoenzymes of higher isoelectric point, showing activity with other substrates such as 3,4-dichloronitrobenzene, did not bind. These isoenzymes did bind to a sulfobromophthalein-glutathione conjugate immobilized on agarose and could be eluted with 5 mM sulfobromophthalein at pH 7.4. The immobilized glutathione resin bound rat liver glutathione S-transferase subunits from all three molecular weight classes.     

Acetylcholine in the choline-deficient housefly, Musca domestica

The distribution of choline metabolites has been studied in the nervous system of housefly larvae reared on diets containing various amounts of choline or on diets containing either β-methylcholine (β-MCh) or N-dimethylethylcholine (DMECh). Adult houseflies were obtained from larvae reared on diets containing at least 4·6 times the level of choline in the basic diet or on diets containing the two choline analogues. The distribution of choline metabolites has been studied in the heads and thoraces plus abdomina of these adults.

Acetylcholine was found to be concentrated in the larval nervous system where it was synthesized preferentially to other choline metabolites when the amount of choline available to the insect was severely restricted. The acetylcholine content of the adult was between six and nine times that of the larval nervous system and approximately 70 per cent was concentrated in the head. The amount of acetylcholine present in flies obtained from larvae fed on diets containing the lowest amount of choline which allowed adults to develop was 230 pmole/insect. Flies obtained from larvae fed on diets with added β-MCh contained 150 pmole of acetylcholine/insect but no detectable acetyl-βMCh. Flies obtained from larvae fed on diets with added DMECh contained 43 pmole of acetylcholine and 127 pmole of acetyl-DMECh/insect.

It is concluded that the two choline analogues spare the choline requirement of the housefly by two different mechanisms. β-MCh displaces choline from the lipids of non-nervous tissue making more available for uptake into the nervous system where sufficient acetylcholine is synthesized for adult development. DMECh is a complete replacement for choline; the acetyl-DMECh acting as a neurotransmitter in place of acetylcholine.     

A complex glutathione transferase gene family in the housefly Musca domestica

In most metazoans, the glutathione S-transferases (GST) are encoded by gene families, and are used to detoxify xenobiotics. We describe the structure of genomic loci coding for the GSTs in the housefly that have been implicated, by both genetic and biochemical means, in mediating insecticide resistance. In earlier work, we showed that one of the theta-class enzymes, MdGST-3, is overproduced in resistant flies and degrades certain insecticides. We used a fragment from a cDNA clone of MdGST-3 as a probe to screen a housefly genomic DNA bank in phage λ. This probe detected multiple gst loci. Genes for GSTs were found in five different, nonoverlapping λ clones, three of which carry multiple, closely linked gsts. Multiple genes for both MdGST-3 and MdGST-4 were found; some of which have introns in their 5′ untranslated regions. In adults, the only MdGST-3 enzymes that are expressed are encoded by the intron-free genes. A new theta-class GST (called MdGST-5) was also discovered. Fusion genes comprising 5′ MdGST-3 sequences and either MdGST-4 or MdGST-5 sequences in their 3′ halves were encountered at three separate loci. The genes described here are found in both the ancestral sensitive strain and the insecticide-resistant strains. Received: 28 June 1996 / Accepted: 23 April 1997     

Fine structure of the Malpighian tubules in the housefly, Musca domestica

The epithelium of the Malpighian tubules in the housefly is comprised of four distinct cellular types. Type I cells are characterized by the presence of intimate associations between infoldings of basal plasma membrane and mitochondria. On the luminal surface, cytoplasm is extended into microvilli which contain mitochondria. Membrane-bound vacuoles in the cytoplasm seem to progressively accumulate granular material. Type II cells have dilated canaliculi. Microvilli lack mitochondria. The Type III cell has not been reported previously in Malpighian tubules. It has very well-developed granular endoplasmic reticulum which contains intracisternal bundles of tubules. Cytoplasm contains numerous electron dense bodies. Type IV cells occur in the common duct region of the Malpighian tubules. Mitochondria do not extend into the microvilli.     

Diel changes in DDT absorption and breakdown rates and respiratory rhythm in the housefly, Musca domestica

Absorption rates in three strains of housefly show little variation when DDT is applied at different times during the 24-hr cycle. Rates of breakdown of DDT to DDE, however, vary significantly at different times of the day with most rapid breakdown at 05.00–05.30 and 15.00 hr, corresponding to diel peaks in oxygen consumption. The pre-dawn peak breakdown rates occur at the times at which the flies were found, previously, to be most susceptible to DDT, thus this change in susceptibility must have other causes.

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Zusammenfassung Die Geschwindigkeit, mit der DDT von drei Stubenfliegenrassen absorbiert wurde, zeigte nur wenig Verschiedenheit, wenn es zu verschiedenen Zeiten im Verlauf des 24-h-Tages appliziert wurde. Die Rate des Abbaus von DDT zu DDE variierte dagegen im Lauf des Tages, sie war am grössten um 5.00 bis 5.30 Uhr und um 15.00 Uhr, entsprechend den Tagesspitzen des Sauerstoff-Verbrauchs. Das erste Abbaumaximum (vor Tagesanbruch) fiel zusammen mit der Zeit, in der die Fliegen höchste Empfindlichkeit gegenüber DDT zeigten (vgl. vorige Veröffentlichung). Diese tageszeitspezifische Empfindlichkeit liess sich demnach keineswegs mit einer (geringeren) Abbaugeschwindigkeit in Zusammenhang bringen.

     

Linkage relationships of several insecticide resistance factors in the house fly (Musca domestica L.)

Summary Strains of the house fly resistant to parathion, malathion, Isolan, and DDT were crossed with a susceptible strain which carried the recessive marker, stubby-wing (stw). F₁ populations were composed of normal-winged insecticide-resistant flies. When the F_1's were backcrossed to the stw parent, the resultant normal-winged progeny were resistant and the stw progeny were susceptible. Thus the major factors for resistance in the strains studied were all located on the same chromosome.Similar cross-over ratios were observed with parathion and malathion-resistant strains and populations of resistant stw flies were established. Low ali-esterase, characteristic of the parent resistant strains, was also present in the resistant stw strains.No crossing-over occurred between stw and DDT-resistance. Resistance to DDT, present in the malathion-resistant strain, was not introduced with malathion-resistance into the stw strain. Therefore, the factors for DDT and malathion resistance, although linked, are genetically distinct.

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Zusammenfassung Stubenfliegenstämme, die Resistenzfaktoren gegen Parathion, Malathion, Isolan^R (1-isopropyl-3-methyl-5-pyrazolyl dimethylcarbamate) und DDT tragen, wurden mit einem anfälligen Stamm gekreuzt, der das rezessive Markierungsgen, stummelflügelig (stw), aufweist. Die F₁-Populationen jeder Kreuzung bestanden aus normalflügeligen, insektizidresistenten Fliegen. Wenn die F₁-Fliegen mit den stw-Eltern rückgekreuzt wurden, waren die normalflügeligen Nachkommen resistent gegen alle Insektizide und die stw-Nachkommen anfällig. Also sind die Hauptresistenzfaktoren der untersuchten Stämme alle in dem gleichen Chromosom lokalisiert.Für die Faktoren, welche die Resistenz gene Parathion und Malathion kontrollieren, wurden ähnliche crossing over-Verhältnisse beobachtet, und es wurden Populationen von stw-Fliegen aufgebaut, die resistent gegen diese Insektizide sind. Niedrige Ali-Esterase-Aktivität, die für die parathion- und malathionresistenten Ausgangsstämme charakteristisch ist, war auch in den resistenten stw-Stämmen vorhanden, so daß das Gen für niedrige Ali-Esterase-Aktivität identisch oder eng mit den Resistenzgenen verbunden ist.Eine unzulängliche Trennung zwischen anfälligen und isolanresistenten Fliegen schloß die Möglichkeit der crossing over-Messung des Faktors für Isolanresistenz aus. Zwischen stw und DDT-Resistenz wurde kein crossing over beobachtet. Hohe Kreuzungsresistenz gegen DDT, die in einem malathionresistenten Stamm vorhanden ist, wurde mit dem Faktor für Malathionresistenz nicht in den stw-Stamm eingeführt. Die Faktoren für die Resistenz gegen DDT und Malathion sind demnach genetisch unterschiedlich.

     

Identification and cloning of a key insecticide-metabolizing glutathione S-transferase (MdGST-6A) from a hyper insecticide-resistant strain of the housefly Musca domestica

Strains of the housefly, Musca domestica, highly resistant to organophosphate (OP) and other insecticides are known because they overproduce glutathione S-transferases (GSTs). Previous work has shown that overproduction in these strains involved numerous isozymes with glutathione conjugating activities (Pesticide Biochem. Physiol., 25 (1986) 169; Mol. General Genetics, 227 (1991) 355; J. Biol. Chem., 267 (1992) 1840; Mol. General Genetics, 245 (1994) 236; J. Mol. Evol., 43 (1996) 236). The current work describes the purification and identification of a M. domestica GST isozyme (pI 7.1) broadly specific for substrates from a housefly strain, Cornell-HR, that is highly resistant against OP-insecticides, and the isolation of two new MdGST genes using the antibody made against it. This isozyme, which was identified from amongst more than 20 isoelectric forms of GSTs of the same subunit size, was highly active for conjugating GSH to the model substrate 3,4-dichloronitrobenzne (DCNB). When expressed in Escherichia coli, one of the cloned GSTs, MdGST-6A, produces an enzyme that conjugates glutathione to the insecticides methyl parathion and lindane. On indication that it was the most active isozyme toward several xenobiotics among several MdGSTs tested, we advance the notion that MdGST-6A probably plays an important role in M. domestica Cornell-HR's resistance towards OP-insecticides. MdGST-6A and a second closely related one found in this work, MdGST-6B, are members of the traditional insect class I family (theta-class) and share the greatest homologies with a cluster of Drosophila GSTs on locus 55. In addition to having the unusually broad substrate specificity, the sequence of the new group of enzymes reveals that it has a highly diverged hydrophobic motif in its active site as compared to other class I GSTs from insects.     

Glutathione and glutathione S-transferases in the salmonella mammalian-microsome mutagenicity test

Levels of the tripeptide glutathione (GSH) and the activity of glutathione S-transferases were investigated in S9 fractions of rats and mice and in Salmonella typhymurium tester strains TA1535, TA100, TA1538 and TA98. The S9 and Salmonella typhimurium tester strains had high levels of glutathione. Compared with S9, the activity of GSH S-transferases was lower in the bacteria. However, electrophiles such as 1-chloro-2,4-dinitrobenzene (CDNB), diethyl maleate and styrene oxide were effectively bound to bacterial GSH.

The mutagenicity of the direct mutagen CDNB was drastically lowered in presence of S9 fractions but not in presence of microsomes. A comparable decrease was obtained when microsomal supernatant, which contains GSH and GSH S-transferases, was added to the microsomes. Addition of GSH in excess completely abolished mutagenicity of CDNB. These results demonstrate that the conjugation of electrophiles with GSH mediated by the S9 fraction or the bacterial tester strains represents an important detoxication mechanism which may influence the results obtained with the Salmonella typhimurium mammalian-microsome mutagenicity test.     

The sibling species Drosophila melanogaster and Drosophila simulans differ in the expression profile of glutathione S-transferases

Two major forms of glutathione S-transferase are known in Drosophila melanogaster: GST D and GST 2. In the present paper we report the existence of a third major form of glutathione S-transferase in Drosophila simulans. Induction with phenobarbital revealed a different regulation of GST between these species. Despite the fact that these two species are closely related, there was a difference in the expression profile of the enzyme implicated in the detoxification system, suggesting variations in capacity to suit their environment.     

Synergism of organophosphates in Musca domestica and Chrysomya putoria

A series of symmetrical trisubstituted phosphorus compounds was tested as synergists for malathion and certain other organophosphorus insecticides against resistant and normal strains of Chrysomya putoria and Musca domestica. All the compounds synergised the insecticides more effectively against resistant than normal strains. This was especially true with malathion, notably with the C. putoria strain the resistance of which is highly specific for malathion. A 5:1 mixture of the best synergists virtually abolished its resistance. Among the synergists tested, the two best were common to the C. putoria and the two different resistant strains of M. domestica; these two synergists were also outstanding against Culex tarsalis larvae, as reported by other authors.In addition to the synergistic action on malathion, the better compounds were also effective in reducing resistance to other phosphorus insecticides (Dicapthon, parathion, coumaphos) though not quite so effectively. It is concluded that their action is not entirely specific for suppressing carboxyesterase detoxication.

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Synergismus von organophosphaten bei Musca domestica und tChrysomya putoria

Zusammenfassung Eine Reihe symmetrisch dreifach substituierter Phosphorverbindungen wurde als Synergisten für Malathion und gewisse andere Organophosphat-Insektizide gegenüber resistenten und normalen Stämmen von Chrysomya putoria und Musca domestica geprüft. Alle diese Verbindungen verstärkten die Insektizide gegen resistente Stämme wirkungsvoller als gegen normale. Das galt besonders für Malathion, vor allem bei dem C. putoria-Stamm, dessen Resistenz gegenüber Malathion hoch spezifisch ist. Ein 5:1-Gemisch der besten Synergisten hob seine Resistenz praktisch auf. Unter den geprüften Synergisten waren die zwei besten C. putoria und den beiden verschiedenen resistenten Stämmen von M. domestica gemeinsam; diese beiden Synergisten wirkten auch gegenüber Culex tarsalis-Larven ausgezeichnet, wie andere Autoren berichteten.Neben der synergistischen Wirksamkeit für Malathion reduzierten die besseren Verbindungen auch die Resistenz anderer Phosphor-Insektizide (Dicapthon, Parathion, Coumaphos), wenn auch nicht ganz so vollkommen. Es wird geschlossen, daß ihre Wirkungsweise für die Unterdrückung der Carboxyesterase-Entgiftung nicht ganz spezifisch ist.

     

The Hermes transposon of Musca domestica and its use as a mutagen of Schizosaccharomyces pombe

Transposon mutagenesis allows for the discovery and characterization of genes by creating mutations that can be easily mapped and sequenced. Moreover, this method allows for a relatively unbiased approach to isolating genes of interest. Recently, a system of transposon based mutagenesis for Schizosaccharomyces pombe became available. This mutagenesis relies on Hermes, a DNA transposon from the house fly that readily integrates into the chromosomes of S. pombe. The Hermes system is distinct from the retrotransposons of S. pombe because it efficiently integrates into open reading frames. To mutagenize S. pombe, cells are transformed with a plasmid that contains a drug resistance marker flanked by the terminal inverted repeats of Hermes. The Hermes transposase expressed from a second plasmid excises the resistance marker with the inverted repeats and inserts this DNA into chromosomal sites. After S. pombe with these two plasmids grow 25 generations, approximately 2% of the cells contain insertions. Of the cells with insertions, 68% contain single integration events. The protocols listed here provide the detailed information necessary to mutagenize a strain of interest, screen for specific phenotypes, and sequence the positions of insertion.     

Knockdown resistance (kdr) to DDT and pyrethroid insecticides maps to a sodium channel gene locus in the housefly (Musca domestica)

The voltage-sensitive sodium channel is generally regarded as the primary target site of dichlorodiphenyl-trichloro-ethane (DDT) and pyrethroid insecticides, and has been implicated in the widely reported mechanism of nerve insensitivity to these compounds. This phenomenon is expressed as knockdown resistance (kdr) and has been best characterised in the housefly where several putative alleles, including the more potent super-kdr factor, have been identified. We report the isolation of cDNA clones containing part of a housefly sodium channel gene, designated Msc, which show close homology to the para sodium channel of Drosophila (99% amino acid identity within the region of overlap). Using Southern blots of insect DNA, restriction fragment length polymorphisms (RFLPs) at the Msc locus were identified in susceptible, kdr and super-kdr housefly strains. These RFLPs showed tight linkage to resistance in controlled crosses involving these strains, thus providing clear genetic evidence that kdr, and hence pyrethroid mode of action, is closely associated with the voltage-sensitive sodium channel.     

Map position of Aldox (Aldehyde-oxidase) and Adh (Alcohol dehydrogenase) loci on autosome II of Musca domestica L.

The Aldox and Adh structural loci of Musca domestica L. belong to autosome II. They code for the enzymes aldehyde oxidase and alcohol dehydrogenase. Both these enzymes have allelic variants with specific electrophoretic mobility, which, on cellogel, are seen as single bands. The Aldox and Adh loci encompass a large map interval, which includes the morphological markers ar, cm, and car. The recombination frequencies between these five loci indicate the alignment Aldox-ar-cm-car-Adh.

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Localisation chez Musca domestica L. des gènes Aldox et Adh codant les enzymes ald\;ehyde oxydase (AO) et alcool déshydrogénase (ADH)

Résumé Chez la mouche, les enzymes AO et ADH, observées par électrophorèse sur cellogel, présentent toutes 2 des formes alléliques exprimées par des bandes ayant une mobilité anodique propre.Les gènes structuraux Aldox et Adh, codant ces formes, sont liés entre eux et situés sur le chromosome 2. Ils se recombinent avec une fréquence élevée d'interchange; ils sont donc séparés par un intervalle important dans lequel sont compris les caractères visibles ar, cm, car. La fréquence des recombinaisons entre caractères visibles et gènes enzymatiques indique l'ordre suivant sur ce segment du deuxième chromosome de la mouche: Aldox, ar, cm, car, Adh.

     

Male housefly (Musca domestica L.) genital system as a source of mating pheromone

Male volatile mating pheromone is released by male houseflies placed in close proximity to females or in contact with female hydrocarbon extract. The pheromone induces the aggregation of virgin females in the vicinity of the male. Preparations of macerated testicular and ejaculatory duct tissue exert a similar effect, and these organs are, apparently, the source of the pheromone. Such preparations are not attractive to males or mated females. The pheromone also appears to induce female receptivity, since females that are exposed to emanations from male system preparations show a quicker rate of pairing with males. The pheromone effect results from the testes and ejaculatory ducts acting together, as preparations of each of these alone are inactive. Pheromone activity is age dependent and the greatest female reaction is observed when tissues of 6–10-day old males are used.     

Host age, sex, and pathogen exposure level as factors in the susceptibility of Musca domestica to Entomophthora muscae

Adult Musca domestica were exposed to conidial showers from cadavers that were killed by Entomophthora muscae. Of the 4 host ages (14 days, 7 days, 3 days and <1 day), older flies were more likely to die without postmortem signs of infection, while younger flies were more likely to die with signs and produce primary conidia. The youngest fly group also succumbed to infection significantly sooner than older flies. Total mortality did not differ for the 2 sexes, but females died significantly more often with signs in one trial. Males died significantly earlier than did females, but differences were not significant when the sexes were paired by weight. Increased levels of pathogen exposure significantly increased both total mortality and the number of flies dying with signs; it also decreased incubation period. The potential significance of these findings in the epidemiology of the disease is discussed.

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Zusammenfassung Die Mortalitätsrate von Musca domestica nach Behandlung mit Entomophthora muscae Sporen wurde in Laborversuchen in Abhängigkeit von Wirtsalter,-geschlecht und Expositionsdauer festgestellt. Fliegen verschiedenen Alters (14 Tage, 7 Tage, 3 Tage und jünger als 1 Tag) wurden gemeinsam in 237 ml Kartonbehältern gehalten. Die Behälter besassen Netzdeckel, auf denen frisch an einer Entomophthora Infektion gestorbene Fliegen ausgelegt wurden. Auf diese Weise wurden die Versuchstiere 8 Stunden (low exposure) bzw. 48 Stunden (high exposure) lang mit Entomophthora Sporen kontaminiert. Mit Hilfe von am Boden der Versuchsgefässe ausgelegten Objektträgern wurde die relative Menge Entomophthora Sporen festgestellt, der die Versuchstiere ausgesetzt waren. Der Totenfall wurde täglich für die Dauer von 10 Tagen registriert; dabei wurde zwischen Konidienträgern (external signs) und Leichen ohne Konidien (no external signs) unterschieden.Die Versuchsgruppen, die 48 Stunden lang mit Entomophthora Sporen behandelt worden waren, wiesen im Vergleich zu den kürzer behandelten Versuchsgruppen höhere Mortalität und einen höheren Anteil an Konidienträgern auf. Darüber hinaus wurde das Auftreten von Konidienträgern bei den länger behandelten Gruppen früher beobachtet. Im Vergleich zu unbehandelten Kontrollgruppen erhöhte die Behandlung mit Entomophthora Sporen auch die Rate nicht konidientragender Leichen.Bei gleicher Expositionsdauer zeigten Weibchen und Männchen ähnliche Mortalitätsraten. Ein höherer Anteil Konidienträger bei den Weibchen wurde bei einem von zwei entsprechenden Versuchen beobachtet. Konidienträger traten bei Weibchen später als bei Männchen auf. In Bezug auf die Inkubationszeit konnte kein geschlechsspezifischer Unterschied festgestellt werden, wenn Versuchsgruppen mit gleichem Gewicht verglichen wurden. Bei beiden Geschlechtern erfolgte die Bildung von Konidien bei grösseren Individuen später als bei kleineren Individuen.Das Alter der Versuchstiere beeinflusste sowohl die Mortalitätsrate als auch die Häufigkeit und das zeitliche Auftreten von Konidienträgern. Jüngere Fliegen wiesen einen höheren Anteil Konidienträger auf; bei Fliegen mit einem Alter von weniger als einem Tag wurde das Auftreten von Konidienträgern früher beobachtet als bei älteren Individuen.Die untersuchten Faktoren können auch für die Epidemiologie von Entomophthora freilebender Musca domestica Populationen eine wichtige Rolle spielen. In Populationen mit niedrigerem Durchschnittsalter wird sich eine Infektion rascher ausbreiten, da jüngere Fliegen eine kürzere Inkubationszeit aufweisen und auch nach kürzerer Kontaminationsdauer Konidien an den getöteten Individuen ausgebildet werden. Darüber hinaus wird sich eine Infektion dann rascher ausbreiten, wenn die Population mit einer grösseren Sporenmenge infiziert wird, da dann eine kürzere Inkubationzeit und ein höherer Anteil Konidienträger zu erwarten ist.

     

The enzymatic degradation of malathion in organophosphate resistant and susceptible strains of Musca domestica

The metabolic fate of malathion in susceptible and organophosphate resistant strains of houseflies was studied by means of radioisotope-techniques, in vivo and in vitro. The main interstrain difference was the superior ability of the two malathion resistant strains to degrade malathion to its monocarboxylic acid derivative. Attempts were made to characterize the carboxyesterases responsible for this difference with respect to heat stability, pH dependency and organophosphate inhibition, and to estimate their importance in causing resistance by means of cross-resistance and synergism experiments. The relation to, and possible identity of the carboxyesterases with, the malaoxon degrading enzymes previously found in the malathion resistant strains is discussed.

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Zusammenfassung Der Malathionmetabolismus wurde mit Hilfe der Radioisotopentechnik in normal empfindlichen und organophosphatresistenten Stubenfliegenstämmen in vivo und in vitro untersucht Die beiden malathionresistenten Stämme konnten Malathion besser in seinen Monocarboxylsäure-Abkömmling abbauen. Es wurde versucht, die hierfür verantwortliche Carboxyesterase hinsichtlich ihrer Hitzestabilität, pH-Abhängigkeit und Hemmbarkeit durch Organophosphate zu charakterisieren und ihre Bedeutung in der Resistenzbildung durch Kreuzungsresistenz- und Synergismusversuche zu bestimmen. Die Beziehung der besonders in den malathionresistenten Stämmen gefundenen malaoxonabbauenden Enzyme zu den Carboxyesterasen und ihre mögliche Identität mit denselben wird diskutiert.

     

The response of the labellar taste receptors of DDT-resistant and non-resistant houseflies (Musca domestica) to DDT

Two characteristics of DDT-resistance in houseflies (Musca domestica L.), namely, a significantly higher threshold to DDT and the ability to recover from DDT-poisoning, were found to be detectable quantitatively at the level of the neurones of the labellar chemoreceptor hairs.Treatment of the chemoreceptor hairs with DDT resulted in replacement of the normal single impulses by groups of 2 or more. Hairs of resistant (HR) flies showed some recovery from this effect after 10–15 minutes, but those of non-resistant (ES) flies did not.Whole flies topically dosed with DDT solutions were rated for the degree of intoxication shown at various times after treatment. In the resistant strain, flies that survived began to recover from intoxication at about 1 hour, but no recovery was observed in the non-resistant strain.The ratios of equi-effective concentrations or dosages for the two strains were found to be very similar in the chemoreceptor hair and whole fly tests. Up to the onset of recovery they were between 3.9 and 8.5 for the hairs and between 2.0 and 6.0 for the whole flies. After recovery commenced, the ratios increased to more than 15 for both the hairs and the whole flies. It was concluded that the resistance of strain HR flies is fully expressed at the level of the chemoreceptor hairs on their labella, and is mainly due to the ability to recover from DDT-poisoning.

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Zusammenfassung Die Wirkung von DDT-Behandlung auf die elektrische Reaktion der Neuronen von chemorezeptorischen Labellenhaaren bei Reizung mit Kochsalz wurde bei Stubenfliegen (Musca domestica L.) eines DDT-resistenten (HR) und eines nichtresistenten Stammes (ES) untersucht. Das Eintauchen einzelner Rezeptorhaare in wässrige DDT-Suspensionen vor der Reizung und Registrierung der Neuronenaktivität durch einen Kochsalz-Elektrolyten bewirkte den Ersatz der normalen Einzelimpulse durch Gruppen von 2 oder mehr Impulsen bei beiden Stämmen. Bei dem resistenten Stamm steigerte sich die Reaktion — gemessen am mittleren Grade der Impuls-Vervielfachung — während der ersten 10–15 Minuten; danach zeigte signifikante Abnahme der Vervielfachung die Erholung der Neurone von den DDT-Wirkungen an. Bei nichtresistenten Fliegen zeigten die Neurone keine signifikante Erholung. Bevor die Erholung deutlich wurde, waren die Chemorezeptorhaare resistenter Fliegen — gemessen an der Konzentration der angewendeten Suspension — 3,9 bis 8,5mal weniger empfindlich gegen DDT als diejenigen der nichtresistenten Fliegen. Das Verhältnis steigerte sich infolge der Erholung in den nächsten 15 Minuten auf mehr als das 15fache.In einer Paralleluntersuchung der DDT-Vergiftung ganzer Fliegen der beiden Stämme wurden einzeln begiftete Fliegen nach dem Grade der Vergiftung geordnet, den sie zu verschiedenen Zeiten nach der Behandlung aufwiesen. Alle nichtresistenten Fliegen, bei denen deutliche Vergiftungserscheinungen auftraten, wurden zunehmend mehr beeinflußt und starben. Es überlebten nur diejenigen, bei denen klare Anzeichen einer Vergiftung zu keinem Zeitpunkt auftraten. Die resistenten Fliegen waren zu einer völligen Erholung von fortgeschrittenen Stadien der Vergiftung fähig. Bei Dosen, die einen gewissen Grad der Erholung gestatteten, stieg in Gruppen resistenter Fliegen der mittlere Grad der Vergiftung nur während der ersten Stunde an, dann fiel er wieder, sobald sich einige Fliegen erholten. Die Relationen — für die beiden Stämme — gleich giftiger Dosen zeigten, bevor die Erholung in Erscheinung trat, daß die resistenten Fliegen 2- bis 6mal weniger empfindlich gegen DDT-Begiftung waren als die nichtresistenten. Erholung der resistenten Fliegen vergrößerte die Relation auf das mehr als 15fache.So ergaben die Untersuchungen an Chemorezeptorhaaren und an ganzen Fliegen sehr ähnliche Werte für die relative Empfindlichkeit der Stämme gegenüber DDT-Begiftung und für das große Ausmaß, in welchem die Widerstandsfähigkeit der Fliegen des HR-Stammes von der Fähigkeit zur Erholung von DDT-Vergiftungen abhängt. Es kann gefolgert werden, daß die Resistenz der Fliegen des HR-Stammes gegenüber DDT vollständig ausgedrückt werden kann auf dem Niveau der chemorezeptorischen Haare auf ihren Labellen. Es wird vermutet, daß Dehydrochlorierung des DDT, welche vermutlich den Mechanismus der Erholung gestattet, auch mit geringerer Sensibilität dieses Stammes gegenüber DDT-Vergiftung in Verbindung gebracht werden kann.