Classifications

• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
  • A01N31/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic oxygen or sulfur compounds
  • A01N31/02—Acyclic compounds
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
  • A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
  • A01N25/002—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing a foodstuff as carrier or diluent, i.e. baits
  • A01N25/006—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing a foodstuff as carrier or diluent, i.e. baits insecticidal
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
  • A01N35/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical
  • A01N35/02—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical containing aliphatically bound aldehyde or keto groups, or thio analogues thereof; Derivatives thereof, e.g. acetals
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
  • A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
  • A01N37/02—Saturated carboxylic acids or thio analogues thereof; Derivatives thereof
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
  • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
  • A01N63/20—Bacteria; Substances produced thereby or obtained therefrom

Definitions

• the present invention concerns attractant traps, lures and baits for fruit flies belonging to the family Tephritidae.
• the invention also relates to methods for attracting and controlling these insects.
• a "bait” comprises an attractant material, especially a food substance, and usually includes a pesticide.
• a “lure” may or may not include a pesticide, and often indicates that a sexual pheromone is used as the insect attractant.
• the term “bait” is assumed to include the term “lure”, and the presence or absence of a pesticide is specifically indicated.
• the present invention is directed to attractant lures and baits for insects of the family Tephritidae, namely fruit flies.
• fruit flies is used to indicate all flies belonging to the family Tephritidae (Diptera), such as Daeus dorsalis (Oriental fruit fly), Daeus tryoni (Queensland fruit fly), Daeus cucurbitae (Melon Fly), Ceratitis capitata (Mediterranean fruit fly), and fruit flies of the genera Rhagoletis and Anastrepha, for example. Insects such as fruit fly cause considerable damage to agricultural crops throughout the world. Spraying and similar methods of applying the crops with pesticides are the most commonly practiced methods of controlling these insect pests.
• lures and baits rely on the principle of providing an attractant for a particular insect pest.
• the attractant can be of food substance, or a sex pheromone, for example.
• the mechanism of the attraction is primarily chemical, although tactile and visual stimuli created by the attractant can also be important.
• the attractants are sometimes used to attract and catch insect pests in traps, or more generally cause the insect pest to come in contact with poisons thereby killing or damaging it, as well as controlling the insect population as a result.
• the use of traps also enables the measurement of population densities, which is important for timing spray applications of insecticide, for example.
• Tephritid fruit flies cause extensive damage to horticultural crops in almost all horticultural districts throughout the world. Damage to fruit and vegetables is caused by fruit fly larvae which tunnel through fruits and in some cases through stem tissue, spreading bacterial rot as they go. The bacteria, which are deliberately introduced into the plant host tissue by the adult female fruit fly along with her eggs, multiply at the expense of this plant tissue. The larvae then feed on the resultant bacterial "soup" inside the host tissue which is soon rendered unfit for human consumption.
• Adult fruit flies, on the other hand, and in particular the females feed on naturally occurring leaf surface saprophytic bacteria. These latter bacteria may be, but need not always be, the same species which the female r e-gurgitates for inoculation into plant tissue with her eggs .
• Cue-lure is 4- (p-acetoxyphenyl)-2-butanone, anisylacetone is 4- (p-methoxyphenyl)-2-butanone, Willison's lure is 4-(p-hydroxyphenyl)-2-butanone, methyl eugenol is O-methyleugenol and trimedlure is t-butyl-4(or 5)chloro-2-methyl cyclohexane carboxylate.
• a fermenting bait to attract fruit flies. It is thought that fruit flies are attracted to this type of bait by it being a food source.
• a typical fermenting bait comprises a sugar solution to which yeast has been added, and which has been allowed to ferment. This bait has not been found to be particularly effective in attracting fruit flies, and is not commonly used these days.
• Another type of bait is the protein hydrolysate bait discovered by Steiner, and this bait is commonly used at present. In general, a protein hydrolysate is combined with an insecticide and, for example, sprayed on fruit trees.
• Protein-malathion hydrolysate bait sprays are suited to aerial application, and the protein solution is generally harmless to vegetation. Spraying is generally carried out using a high viscosity spray mixture, with large droplet size, in order to ensure the survival of non-target insect species, which will not be attracted to the mixture. (See M.A. Bateman in Drew, Hooper and Bateman, Economic Fruit Flies of the South Pacific Region, Brisbane, 1982) Experiments involving the proteinaceous baits for controlling oriental fruit flies are described in an article by Paul L.
• Gow entitled “Proteinaceous Bait for the Oriental Fruit Fly” in the Journal of Economic Entomology, 47 (1), 153-160, (1954). It is of interest that on page 156, Gow noted that "the attractants in the proteinaceous baits were produced by microbial action on the original protein material". In addition, Gow isolated .various bacterial strains obtained from the protein hydrolysate, and tentatively identified one bacterial colony as belonging to the genus Proteus. However, no further identification of this bacterial species was provided in the article or subsequently, and in the experiments the proteinaceous baits were colonised by random bacteria from the general environment.
• the present invention comprises a bait for fruit fly comprising one or more live bacteria species of the family Enterobacteriacae on which the fruit fly feed, and a substrate able to support the growth of said bacteria species, and an agent toxic to the fruit fly.
• the agent toxic to the fruit fly may be a pesticide for example.
• a suitable such pesticides is malathion.
• Other pesticides are those known in the art to kill or damage fruit fly.
• the toxic agent may be a suitable fruit fly pathogen; for example a bacterium or virus which can kill the insect or otherwise be detrimental to it. As well as killing the fruit fly, the agent may damage, or destroy the reproductive abilities of the fly, or otherwise act to control the fly population. More than one pesticide or pathogen can be included. The chosen agent should not be harmful to the bacterial attractant.
• the invention also concerns a storage stable pre-mix for preparing the above described fruit fly bait.
• One form which these premixes can take is to combine a dried powdered substrate with dried and inactive live bacteria.
• a dried bacteria/dried substrate composition can be stored for long periods. When it is desired to use the composition, it merely needs to be moistened and left for a period for the bacteria to begin growth again.
• the pre-mix bacterial culture is dried by freeze drying.
• the pre-mix may comprise a dried powdered substrate with inactive bacteria, without the toxic agent required for the bait.
• the toxic agent for example a pesticide
• another pre-mix comprises the dried, inactive bacteria, preferably mixed with any desired additives such as dried CMC, glucose or urea.
• the substrate, and the pesticide are added when the bait or attractant is made up.
• the pre-mix may also be part of a kit, wherein the inactive bacteria is provided together with the substrate, either dried or ready for use. If the pre-mix is to be used as an attractant in a trap, for example, the pesticide or other toxic agent, need not be included in the attractant composition. When the pre-mix is made up, and moistened, the bacteria will become active, and begin growing on the substrate.
• the bacteria and substrate or storage stable pre-mix may include the pesticide, or the pesticide may be added at the time the bait is applied in the field.
• Any substrate conducive to the growth of the preferred bacterial inoculum may be used. Acid hydrolysates of proteinaceous material, such as soybean or wheat, or alternatively autolysates, may be used, but the preferred substrate is a yeast autolysate, extract, or powder. In dried form such a substrate may, for example, contain:- Moisture 4.1%
• non-hydrolysed material such as soy meal or pea flower may be used as a substrate, in which case the preferred inoculum inself makes the initial breakdown of the substrate.
• attraction to fruit flies may be delayed about a week, depending on temperature and other conditions, following inoculation.
• baits should be used in traps rather than as foliar applications.
• the substrate may contain one or more additives to promote, retard, or regulate bacterial growth in a manner consistent with maximum attractancy to fruit flies.
• Suitable additives to enrich the nutrient content of the substrate are, for example, glucose, other sugars, peptones, amino acids, urea, ammonium salts, potassium hydrogen phosphate salts, sodium thiosulphate, or B-vitamins.
• Other additives conducive to bacterial activity and the attractancy of the bait are those which retard dehydration or facilitate rehydration and the leakage of nutrients from within the applicable plant leaf or other surface to the attractant bait composition thereon.
• These water regulating materials may for example be hydrophilic gels or colloids, agars, extenders or food thickeners, or they may be organic films which present a barrier to evaporation.
• Sodium carboxy methyl cellulose and hydroxy propyl cellulose are examples of suitable extenders.
• Miller's "Nu-film 17" is an example of an anti-evaporative film.
• the pH of the attractant composition may be altered with suitable agents.
• the substrate may be modified by additives or other means to selectively favour the growth of the preferred bacterial inoculum over and above the growth of volunteer species of microorganism already present on applicable plant surfaces, or in the general environment of those surfaces.
• sodium taurocholate bile salt
• the substrate will be more or less used up by random less attractant or non-attractant bacterial species or strains from the applicable surface or general environment, and the attraction of the bait will be proportionately spoiled.
• the substrate may be altered by the addition of what in medical terminology are called chemotherapeutic agents or antibiotics, some of which are bacteriocidal and others bacteriostatic, different examples having different spectra of activity against some but not other bacteria.
• chemotherapeutic agents or antibiotics some of which are bacteriocidal and others bacteriostatic, different examples having different spectra of activity against some but not other bacteria.
• Klebsiella aeruginosa a very similar bacterium to K.pneumoniae isolated below from the fruit fly Dacus dorsalis, is resistant to the Sulphonamides.
• K. aeruginosa were used as the inoculum, the addition of a sulphonamide to the bait would eliminate competition, not only from, for example, leaf surface Bacillus species, but also many other competing
• Example 1 Enterobacteriaceae.
• uninoculated baits using the preferred yeast autolysate substrate were of low attractancy during the first day, and thereafter, or sooner, to varying degrees caught up with the inoculated baits in terms of the number of flies caught. It is believed that the degree to which they "caught up" reflects the attractancy of the volunteer species.
• Example 1 treatments No. 4 and No. 9 may be compared in this regard.
• additives of these nutritive, regulatory, selective and other kinds are well understood by persons skilled in the art of the present invention, and commonly known additives can be used in the invention.
• Additives may be incorporated into the attractant composition, or into any storage stable pre-mix preparation referred to above.
• the protein autolysate and hydrolysate substrate has a low salt (NaCl) content.
• Protein autolysate and hydrolysate substrates which are readily available can contain added salt, and are known as “high salt” substrates, or contain small amounts of added salts, or no added salt, and are known as “low salt” or “salt free” substrates.
• a high salt substrate will contain approximately more than about 5 % w/v salt and often between about 12 and about 16 % w/v salt.
• Low salt autolysates will typically contain about 1.5 % w/v salt, which is the natural amount salt of present in the protein autolysate without any additional salt being added.
• Acid hydrolysates tend not to support strong bacterial growth and their comparatively low attractancy to fruit flies may possibly be due to de-naturing of the plant protein ingredient during acid hydrolysis rather than to their salt content.
• the bacteria species present in the attractant composition or bait be a single species. Two or more species can be included in a single composition or bait, but in this case the two or more strains will compete with one another, and one will predominate in the long run.
• Preferred bacteria species are of the genera: Erwinia, Citrobacter, Providencia, Klebsiella, Serratia or Salmonella.
• certain species of genus Proteus are also preferred.
• Earlier experiments by Gow isolated an unidentified bacteria of genus Proteus, and so no novelty is claimed for all bacteria of this genus.
• particular species of Proteus which have now been identified and found to be attractants for fruit fly are claimed.
• Preferred bacteria species are chosen from Erwinia herbicola, Citrobacter freundii, Citrobacter diversus,
• Providencia rettgeri Klebsiella oxytoca, Klebsiella pneumoniae, Proteus vulgar is. Salmonella cholerae-suis, or Serratia liquefaciens.
• Providencia rettgeri is synonymous with Proteus rettgeri.
• the bacteria of genus Salmonella are harmful to humans and animals, and are therefore not particularly preferred for use in the baits and traps of the invention.
• bacteria of species Enterobacter cloacae which is a contaminant and which is not considered as one of the attractant bacteria of the invention.
• the bacteria of the invention are leaf and fruit surface saprophytes belonging to family Enterobacteriaceae. Such bacteria grow strongly in a manner attractive to fruit flies where the plant surface substrate is rich in hydrolysed plant proteins, or in protein precursors such as amino acids and peptides.
• bacteria are generally obtained by catching wild fruit flies of the species in question usually while they are feeding, and, using sterile dissection techniques, removing crop and stomach.
• the crop, in particular, and the stomach contains, among other micro-organisms, the required inoculum.
• Enterobacter cloacae a common inhabitant of the lower gut of fruit flies, presents an example of a non-required contaminant which may be found in the crop together with the ingested bacterial food.
• the crop and stomach contents may be spread on the surface of a suitable medium, such as a peptone yeast extract (PYE) agar (10g peptone, 5g yeast extract, 5g NaCl, 15g agar, 1,000ml distilled water) and incubated at a suitable temperature.
• PYE peptone yeast extract
• the most common colony types found in a series of plates from a number of crops and stomachs are selected and pure cultures obtained.
• the attractant composition consists of a substrate with or without the hereinabove described additives, and a bacterial inoculum.
• the so called low salt yeast autolysate (LSA) produced by Mauri Foods of Toowoomba, Queensland, is a good example of a preferred substrate, although this Mauri Foods product does require refrigeration during storage.
• a powdered dried form of LSA could be stored for long periods at normal temperatures. Most preferably 50mls of LSA or other substrate, or 25g of protein solids, are added to each litre of water to make the bait.
• an insecticide can be added to the inoculated substrate.
• Malathion as the 103% emulsifiable concentrate, at the rate of approximately 0.1% is one example of a suitable pesticide.
• a food thickener or extender such as carboxy-methyl-cellulose (CMC) may be added to thicken the inoculated substrate.
• CMC carboxy-methyl-cellulose
• a solution of CMC is preferred which will make the solution sufficiently thick without impeding the application onto foliage through a spray nozzle.
• “Spots” or “squirts” of this bait preparation may consist of about 100ml each, and is most preferably mainly applied to the undersides of green foliage. Aerial application to the upper surfaces of the foliage from light aircraft or helicopters, is also satisfactory but in this case the bait is preferably not dispersed with a fine spray nozzle.
• “spots” may be applied on a regular grid spacing to combat sedentary fruit flies (M.A. Bateman op. cit. 1982) . where the target population of pest fruit flies is sexually immature, and therefore less sedentary, the distance between "spots" can be increased. This may apply in eradication programmes where teneral, or immature, flies are moving away from a locus of infestation.
• spots are applied and calico sheets may be spread beneath each spot to monitor the kill.
• white sheets of apprximately 2 square metres in area below a "spot" of 1 square metre will provide the required result, of course provided that ants, frogs, skinks, and so forth are not permitted to remove the catch.
• This inoculated attractant preparation can be used in a trap, with or without insecticide depending on the mode of operation of the trap.
• fruit flies are not particularly attracted to protein hydrolysates made by acid hydrolysis when such are applied in wicks or traps
• fruit flies are strongly attracted to inoculated substrates, in accordance with the invention, applied in such devices.
• inoculated attractant preparations are attractive to fruit flies even when applied to unusual target surfaces, such as fence posts, brick walls, or telegraph poles for example.
• the non-inoculated preparations of the prior art are scarcely attractive at all on these non-living surfaces, and must be applied to green leaves and fruit which fruit flies normally frequent and where volunteer inocula may be present.
• a further aspect of the present invention relates to baits and attractants for fruit flies involving chemical compounds chosen from those naturally produced in bacterial volatile emissions.
• chemical compounds chosen from those naturally produced in bacterial volatile emissions.
• Several volatile compounds were identified by Lee, Smith and Freeman ("High-resolution gas chromatographic profiles of volatile organic compounds produced by microorganisms at refrigerated temperatures",
• 2-butanone was tested and found to act as an attractant for fruit fly.
• Other compounds namely phenyl acetate, butan-1-ol and isopentanol were also found to be attractive to fruit fly. It was found that the combination of these compounds functioned better than the individual compounds.
• 2-butanone was particularly effective as an attractant among the compounds tested under laboratory conditions . Further details are provided. in Example 1. If desired, these attractant compounds, (preferably stabilized chemically by adsorption on to inert powders such as talc, or by microencapsulation, for example) may be added to the bacterial lures discussed previously.
• the attractant compounds are very volatile, it is preferred to use them in a controlled release formulation; for example, microencapsulated.
• Fruit fly can be caught in traps using the attractant compositions or baits of the invention. Any of the commonly known traps for catching insects can be used. Merely some examples of these which are used for catching fruit flies are the Steiner trap, the Israeli trap, the Bateman trap, the Jackson trap, or the McPhail trap. These traps are discussed in more detail in Drew, Hooper and Bateman, op.cit. (1982), on pages 129 to 133. Traps can catch fruit fly by having an opening which allows the flies to easily enter the trap but which restricts them from leaving it. Otherwise, the trap may incorporate a sticky surface to hold the fruit flies, and this restricts the fly from leaving the trap.
• the preferred attractant bacteria for use as the bacterial inoculum in the invention is Erwinia herbicola.
• a strain of Erwinia herbicola which is particularly effective has been deposited with the American Type Culture Collection (ATCC) at Rockville, Maryland, the United States of America, under the Applicant's reference number "BTA 751". The ATCC deposit number is lodged on
• Example 1 Synthetic bacterial volatiles
• a number of Steiner traps were set up at equal spaces around a field cage. Dacus tryoni were released into the room after thimbles, with a pin hole for release of the attractant, were placed in each trap. The thimbles contained the different chemicals with one empty as a control. No insecticide was used and each trap was examined at 60 minute intervals throughout the day. The contents of each trap were noted - the flies were counted and sexed - and released back into the room. Cue-lure was also included for comparison purposes. The totals for each chemical were:
• Control 50 31 38 81 A s ignif icant feature is the number of females caught by 2-butanone as the lure in comparison with cue-lure . By catching a s ignificant number of females , 2-butanone f unctioned much better than cue-lure, which hardly ever catches females at all.
• the attractant or bait mixture may be used to attract fruit flies into a plastic or glass or other trap containing the attractant mixture.
• the bait may be applied to green foliage in orchards, as, for example, a controlled release formulation of the preferred compounds.
• the mixture may be used in association with a suitable insecticide such as malathion, dichlorvos, or pyrethroids.
• the insecticide may also be given controlled release formulation either separately or after mixing with the attractants.
• the constituents of the attractant mixture, their relative concentrations, and the methods of distributing the attractant mixture can be varied to suit the particular applicable conditions.
• the attractant mixture can provide an effective lure for the damage causing female fruit flies which are not, if barely, attracted to the known synthetic lures .
• Example 2 Field attractancy of baits Five experimental trials were conducted against Dacus tryoni and D.neohumeralis in abandoned Guava groves in South East Queensland during autumn 1984. 100ml spots of various preparations, using different recipes and different bacteria were tested under these field conditions. In each trial, 2-square-metre calico sheets were suspended beneath each spot to monitor the catch.
• An LSA substrate should preferably not be autoclaved or otherwise heated before use.
• a liquid LSA substrate should preferably not be stored without refrigeration.
• An inoculated bait using autolysate or hydrolysate substrates should preferably be held for as short a time as possible before application.
• an inoculum is added as a broth, a small volume (20ml/litre) is preferred.
• Erwinia herbicola proved a superior species of inoculum to Providencia rettgeri under these field conditions.
• Escherichia coli proved less attractive, and Pseudomonas putida of family Pseudomonadaceae much less attractive under the same field conditions.
• sample 1 The bacteria in sample 1 were isolated from a laboratory culture of fruit flies that had been in a field cage at sea level, outside this species normal range, for several weeks.
• Example 5 Comparative attractiveness of bacteria An experiment was conducted to determine the comparative attractancy of three bacterial isloates obtained from fruit flies. In a 14 cubic metre field cage containing 600 sexually mature D. tryoni, 12 Golden Delcious apples were coated with inoculated baits and suspended at head height. The baits contained 6% Mauri Foods' low salt yeast autolysate, 1.5% glucose, 2% sodium CMC, and one of the three inocula. Four apples were assigned at random to each bait. The number of flies feeding on each apple were then counted at 30 minute intervals. At each count the flies were removed and each apple rotated two stations clockwise in the cage. The following results were obtained.
• Example 6 Nutrient additives An experiment was designed to measure the effect of added sugar on the attractancy of a bait using Mauri Foods low salt yeast autolysate (LSA) as the substrate.
• LSA Mauri Foods low salt yeast autolysate
• the inoculum was Providencia rettgeri.
• the two inoculated LSA treatments included 2 1/2% CMC.
• 3% glucose was added, and this treatment was referred to as "Queenslure II" or "Q-II”.
• Q-II glucose
• LSA without additives or innoculum and also the two commonly known acid hydrolysate fruit fly baits "Lanes Protein Hydrolysate” and “Staley's P.I.B.-7" also without additives or innoculum were included in the trial.

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• 1984
  • 1984-10-17 WO PCT/AU1984/000204 patent/WO1985001638A1/en not_active Application Discontinuation
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