Rôle of labelled dietary fatty acids and acetate in phospholipids during the metamorphosis of Pieris brassicae

The incorporation of labelled dietary palmitic, linoleic, and linolenic acids into neutral (NL) and phospholipids (PL) during the metamorphosis of Pieris brassicae was studied, and the ability of the fat body to incorporate acetate into PL determined. Thirty-three per cent of total lipid in early fifth instar larvae (minus haemolymph) is PL, while the corresponding value in female 4-day pupae is 13·0 per cent and in the fat body of 4-day pupae 6·3 per cent. Incorporation of label into PL was studied more closely and in all cases the label was recovered from phosphatidylcholine (PTC) and phosphatidylethanolamine (PTE). The label from palmitate was also found in sphingomyelin and possibly phosphatidylserine. Specific activity of PL in the case of palmitic and linolenic acids was greatest in late fifth instar larvae. In early fifth instar larvae on palmitic acid-1-¹⁴C 39·0 per cent of label was in PTC, 52·8 per cent in PTE, and 2·0 per cent in sphingomyelin. In late fifth instar 45·0 per cent was in PTC, 45·5 per cent in PTE, and 6·5 per cent in sphingomyelin, while in 4-day female pupae 45·2 per cent was in PTC, 41·3 per cent in PTE, and 13·5 per cent in sphingomyelin. The label from linolenic acid only varied a little from early fifth instar to 4-day pupae, 51·8 per cent being in PTC and 48·2 per cent in PTE in early fifth instar larvae. The label from linoleic acid is incorporated in fat body PL almost exclusively in PTC and PTE, 55·8 and 43·2 per cent respectively in 4-day female pupae. Injected acetate is distributed after 1 hr between PTC (58·6 per cent), PTE (24·4 per cent), and sphingomyelin (17·0 per cent). It was concluded that the polyunsaturated acids are proportionately more common in PTE than in other PL types, and that the fatty acids of sphingomyelin are mainly those that the insect is capable of synthesizing from acetate. Palmitic acid is desaturated by Pieris to palmitoleic acid and the latter possibly utilized in PTE to compensate for a deficiency of linolenic acid in the artificial diet. No saturation of linoleic or linolenic acid was found. The rates of PL and NL synthesis during development and the rôle of the investigated fatty acids in the biosynthesis of PL are discussed.     

Evidence for selective absorption of polyunsaturated fatty acids during digestion in the tobacco budworm, Heliothis virescens F.

Heliothis virescens F., the tobacco budworm, were reared individually on a semi-defined diet. The 5th and 6th instar larvae digested and absorbed approx. 72% of the available dietary lipid. Analysis of the fatty acid composition of total lipid in the diet and that remaining in the faeces indicated that the polyunsaturated fatty acids, linoleate and linolenate, were preferentially absorbed as compared to saturated and monoenoic fatty acids.     

Lipids and fatty acids of the southwestern corn borer,Diatraea grandiosella

The lipids of the adults and of several immature stages of the southwestern corn borer, Diatraea grandiosella, were studied after they were fed natural corn stalks or artificial diets. Linoleic acid (18:2) was the major fatty acid of the neutral lipids in both the natural and the artificial diets, but aleic acid (18:1) was the principal neutral lipid in all insect stages. Also, linoleic acid and oleic acid were the principal acids in the insect phospholipids of all stages. The content of linoleic acid in the natural diet was also high, but that in the artificial diet appeared to be much too low for insect requirements. Phosphatidyl choline (PC) and phosphatidyl ethanolamine (PE) were the major phospholipids in all growth stages. Thus, in larvae diapausing in the field, the unsaturated fatty acid content of PC was 59·3 per cent, primarily 16:1 and 18:1, and PE was 87·4 per cent, primarily 18:1, 18:2, and 18:3, and the fatty acids in the number 1- and 2-positions of PC were 53·6 and 97·2 per cent unsaturated, respectively. The haemolymph of diapausing southwestern corn borer larvae contained primarily glycerides but also had some PC and PE. Fat body from diapausing larvae contained primarily 16:0, 16:1, and 18:1 in a ratio of 1 : 1 : 2. Thus lipids of the southwestern corn borer do not reflect dietary lipids as closely as do other insects studied.     

Lipid nutrition during larval development of the parasitic wasp, Exeristes

The parasite Exeristes roborator grew rapidly and completed larval development on fatty acid free chemically defined diets. Dietary supplements of palmitoleate, oleate, linoleate, and linolenate were detrimental to parasite development with most larvae dying in the first instar. Palmitate supplements were also toxic, but a small percentage of larvae consistently completed development and survival and development time on diets supplemented with free stearate did not differ significantly from results obtained with fatty acid free diets. Supplements of a mixture of all six free fatty acids were as toxic as the unsaturated free fatty acids. Dietary supplements of the triglycerides, tripalmitin, tripalmitoleate, tristearin, and trioleate had no positive nutritional value for larval growth and development but were not detrimental.Development time was increased when the parasite was reared on fatty acid free diets lacking carbohydrate, but survival was not affected. The parasite, therefore, appears to have the ability to utilize dietary free amino acids as the sole energy source at this stage. Under these nutritional conditions, supplements of triglycerides did not replace the nutritional value of carbohydrate and some of the supplementary triglycerides were detrimental to larval survival.     

Dietary cholesterol requirements of housefly, Musca domestica, larvae.

Quantitative analyses have been made of the dietary cholesterol requirement for the growth of the larvae of Musca domestica. The larvae will not grow on diets to which no cholesterol is added, a few pupae and adults are obtained when the concentration of cholesterol is 0·05 μmol/g of diet, but the concentration has to be raised to 0·36 μmol/g of diet before the maximum numbers of pupae and adults are obtained. Further addition of cholesterol above 0·36 μmol/g diet did not have any significant effect on the weight and growth of the larvae. However, the ratios of the cholesterol to phospholipid fractions recovered from the larvae increased rapidly when the concentration of cholesterol in the diet was raised from 0·05 to 0·56 μmol/g of diet. Above this concentration only a slight increase in the ratios was observed. Larvae reared on diets containing 0·05 μmol cholesterol/g of diet contain only 25 per cent of the cholesterol content of the larvae reared on the diets containing more than 0·28 μmol of cholesterol/g of diet, the cholesterol content being expressed relative to the weight of the larvae,The absence of cholesterol synthesis has been demonstrated in the larvae by feeding [4-¹⁴C] cholesterol. The specific activity of the cholesterol recovered from the larvae is the same as that of cholesterol added to the diet. Irrespective of the cholesterol concentration of the larval diet, approximately 97 per cent of the radioactivity recovered from the larvae behaved as free cholesterol, less than 1 per cent as cholesterol esters and the rest as unidentified ‘polar sterols’. The results are compared with those from similar studies on other insects.     

Ommochrome formation and kynurenine excretion in Pieris brassicae: Relation to tryptophan supply on an artificial diet

The brown-red pigment in the larval epidermis and in the testis of Pieris brassicae was identified as xanthommatin on the basis of solubility, redox behaviour, chromatography, degradation, visible and infrared spectra. In the epidermis, this pigment accumulates during the larval feeding period and disappears rapidly in the wandering stage. Larvae fed an artificial diet produce about half the amount of xanthommatin as larvae fed cabbage. This effect is caused by a lack of dietary tryptophan. Xanthommatin formation is increased by the addition of tryptophan which also increases body weight. At a tryptophan concentration of 0.2 mg per g, however, weight increase is lower than in controls and high mortality is observed. Pieris larvae excrete kynurenine in relation to dietary tryptophan. No measurable amounts are excreted in the last instar on the non-supplement diet. After feeding different quantities of tryptophan, different amounts of kynurenine are excreted only on the day following ecdysis.     

Effects of the parasitization of a braconid, Apanteles, on the blood of its host, Pieris

Parasitization of a braconid wasp, Apanteles glomeratus, of larvae of a common cabbage butterfly, Pieris rapae crucivora, caused changes in differential haemocyte count (DHC), total haemocyte count (THC), and encapsulative capacity against dead eggs of Apanteles in the fourth and fifth instar host larvae.However, no correlation could be found between the number of Apanteles eggs deposited and THC of the middle fourth instar host larvae or between the number of parasitoid larvae and specific gravity of the haemolymph from the late fifth instar host larvae.From the changes in DHC and in THC of both non-parasitized and parasitized Pieris larvae, an increase in the number of plasmatocytes of non-parasitized Pieris larvae in the early fourth instar period was supposed to be due to transformation of prohaemocytes into plasmatocytes, and a low population of plasmatocytes of parasitized larvae in the comparable period was assumed to be due to a suppression of transformation of prohaemocytes by some factor released from the parasitoid eggs.Failure of the parasitized fourth instar Pieris larvae to encapsulate injected dead eggs of Apanteles indicated that the parasitoid embryos were, in some way, actively inhibiting the encapsulation reactions of the host.The increase in THC of the parasitized fifth instar larvae could not be ascribed to a decrease in the volume of host haemolymph. Rather it could be interpreted by a suppression of adhesive capacity of haemocytes in the host haemocoel to tissue surfaces.Reduced encapsulative capacity of the parasitized fifth instar larvae might be attributed either to a depression of the adhesive activity of plasmatocytes resulting from a depletion of energy source for haemocytes in the host haemolymph by parasitization, or from an active suppression of adhesiveness of the plasmatocytes by secretions from ‘giant cells’ (teratocytes) originated from the parasitoid.     

Lipid utilization in adult Pieris brassicae with special reference to the rôle of linolenic acid

Fatty acids of adult Pieris brassicae and the incorporation of dietary linolenic acid-1-¹⁴C into adult (and egg) lipids were analysed 1 and 9 days after ecdysis. Females grown on a leaf diet retained palmitic, palmitoleic, and oleic acids but lost linoleic and linolenic acids during adult life, while males utilized their fatty acids more evenly. On an artificial diet both sexes retained palmitic acid but utilized palmitoleic and oleic acids. In both cases females laid eggs with a high palmitic and oleic acid content.Analysis of thorax flight muscles (artificial diet) revealed that 67·9% of the lipids in 1-day females and 83·6% in 9-day females was phospholipid (PL). During adult life linolenic acid increased in thorax neutral lipids (NL) from 14·6 to 20·0% in females and from 18·5 to 30·0% in males. Males incorporated more linolenic acid-1-¹⁴C especially in fat body and flight muscle PL than females. The majority of this was recovered from phosphatidyl cholines (PTC) in 1-day adults whereas in 9-day adults phosphatidyl ethanolamines (PTE) and another compound, most likely cardiolipin, contained more label (29·0% in PTC, 33·1% in PTE, 34·9% in cardiolipin, and 2·0% in sphingomyelin in the thorax of females). The females also incorporated the label into egg lipids (42·2% in PL, 57·8% in NL). There was recovered from PTC 54·5% of the label in egg PL.Most of the label in thorax NL was found to be in free fatty acids (FFA). The label disappeared from triglycerides during adult life and tended to accumulate in FFA (82·7% in 9-day females) while in diglycerides the label did not vary during adult life (17·2% in 9-day post-emergence females). PTC apparently is a fairly labile PL type which is utilized in muscle whereas PTE and cardiolipin may be more structural in function and accumulated more label from linolenic acid with increasing adult age. Linolenic acid, then, essentially is a structural fatty acid and its rôle appears to be mainly in the structures of flight muscle membranes and organelles.     

Growth and silk formation of silkworm larvae influenced by phytoecdysones

The fourth and fifth instar larvae of the silkworm were reared on artificial diets containing ponasterone A, ecdysterone, and inokosterone. The growth of the larvae and their silk glands, fibroin-synthesizing activity, and silk formation have been investigated. With a diet containing ponasterone A, the fourth instar larvae grew slowly and only a few larvae could ecdyse, while the growth of the fifth instar larvae was disturbed and they died with a darkening of the skin. Ponasterone A also inhibited the growth of the silk glands during the fifth instar. In contrast, the other two phytoecdysones did not greatly influence larval growth. The fourth instar larvae grew rapidly and their ecdysis was advanced with a diet which contained 10 μg of inokosterone/1 g of dry diet. The diet which contained 5 μg of ecdysterone or 10 μg of inokosterone/1 g of dry diet accelerated maturation, while that containing 10 or 20 μg of ecdysterone, or 40 μg of inokosterone, delayed maturation of the fifth instar larvae.Only phytoecdysones caused a decrease in growth of the silk glands in the early half of the instar, and a large amount of phytoecdysones accelerated their growth during the last part of the fifth instar. The fibroin-synthesizing activity was levelled up by feeding ecdysterone and inokosterone, and inokosterone appreciably stimulated activity. Assay of in vitro fibroin synthesis showed that ponasterone A competed with ecdysterone in a stimulative action. Silk formation was much lower in larvae fed the diet containing 5 μg of ecdysterone or 10 μg of inokosterone/1 g of dry diet and was far greater in larvae fed the diet containing 40 μg of inokosterone than in the controls.     

Effects of diet on development of Podisus nigrispinus (Dallas) (Het., Pentatomidae), a predator of the cotton leafworm

Abstract:  The effects of diet on development of Podisus nigrispinus (Dallas) (Het., Pentatomidae) were studied at 25°C, relative humidity of 60 ± 10%, and photoperiod of 14 h light : 10 h dark. Development times of P. nigrispinus nymphs were similar when fed with third or fifth instar larvae of cotton leafworm ( Alabama argillacea Hübner) (Lep., Noctuidae) or Tenebrio molitor L. (Col., Tenebrionidae). When fed with housefly larvae ( Musca domestica L.) (Dipt., Muscidae) or artificial diet, the predator had a longer development time. Independent of diet, instar or sex, the females of P. nigrispinus showed a longer longevity than the males. The total survival of the nymphal stage, on the different diets, varied from 22.46 (fed with housefly) to 77.33% (fed with T. molitor larvae). P. nigrispinus males were heavier when fed with third or fifth instar cotton leafworm larvae than when fed with artificial diet. The weight of the females varied from 37.91 (with artificial diet) to 64.68 mg (with fifth instar cotton leafworm larvae). Independently of the diet, newly emerged females of P. nigrispinus were heavier than the males. Females of P. nigrispinus which were fed with fifth instar cotton leafworm larvae had heavier ovaries than those fed other diets.     

Lipid requirements of the Angoumois grain moth, Sitotroga cerealella

The Angoumois grain moth, Sitotroga cerealella, grew well in an artificial pellet containing about 11 per cent whole wheat germ as its only crude adjuvant. A study of the insect's lipid requirements showed that while dietary wheat germ oil slightly enhanced the larval growth rate it was not required to complete metamorphosis. Normal development without dietary methyl linoleate further confirmed the insect's capacity to mature into an aerial adult without ingesting polyunsaturated fatty acids. A study of the insect's sterol requirements revealed that while a dietary sterol was essential, all free sterols (6) and sterol esters (3) tested permitted equivalent growth and development. Results also showed that larvae did not utilize the sterol relatives, squalene, lanosterol, and cholestane. It was concluded that Sitotroga had a non-specific sterol requirement. Factors in the lipid-free portion of wheat germ, which probably include vitamins, minerals, and amino acids, proved to be essential for normal growth and development, but their exact nature has yet to be determined. The significance of the results in relation to other similar investigations on grain- and plant-feeding insects is discussed.     

亚洲玉米螟幼虫对氮、糖的吸收和利用

本文以不同含氮和含糖量的系列人工饲料为材料,研究了亚洲玉米螟Ostrinia fur nacalis(Guen6e)幼虫对氮和糖的吸收和利用。结果表明：幼虫对饲料中氮的吸收能力随虫龄增加而下降,对糖的吸收能力则随之而增强;5龄幼虫对糖的吸收能力随所食饲料中氮含量的增加而增强,而对氮的吸收则与饲料糖含量关系不大,5龄幼虫对氮和糖的消耗速率均随饲料含糖量增加而加快,但幼虫的氮利用率和虫体含氮量则随之下降;幼虫的氮消耗速率随饲料含氮量的增加而增加,而氮利用率则下降,5龄幼虫的氮利用率比3龄幼虫的氮利用率低,导致虫体含氮量的迅速下降。这些结果说明饲料中氮含量影响幼虫对糖的吸收,高龄幼虫对糖的需要量增加而对氮的需要则较稳定,饲料中糖比氮更重要。     

Differential net food utilization by larvae of Sitophilus oryzae and Sitophilus granarius

The net utilization of a meridic diet by Sitophilus oryzae from the newly hatched larva to the pupa was more efficient compared with that of S. granarius. The artificial diet was highly digestible to both species, but S. oryzae converted 24·1 per cent of the ingested food into body substance compared with 12·4 per cent converted by S. granarius. The mean pupal weight of S. granarius (1·402 mg) was greater than that of S. oryzae (1·012 mg); however, larvae of S. granarius consumed 2·6 times as much food and excreted 8·1 times as much faecal material to obtain that weight. Even though S. granarius consumed the diet at a greater rate, the relative growth rate of S. oryzae was faster. The intracellular symbiotes present in S. oryzae may have a rôle in the species' faster development and more efficient utilization of its food.     

Effects of dietary variation on growth,composition, and maturation of Manduca sexta (Sphingidae: Lepidoptera)

Most studies linking dietary variation with insect fitness focus on a single dietary component and late larval growth. We examined the effects of variation in multiple dietary factors over most life stages of the sphingid moth, Manduca sexta. Larvae received artificial diets in which protein, sucrose, and water content were varied. The relationship between larval size, growth and consumption rates differed significantly across diets. Larvae on control and low-sucrose diets grew most rapidly and attained the largest pupal and adult sizes. Conversely, larvae on low-water and low-protein diets initially grew slowly, but accelerated in the fifth instar and became pupae and adults comparable to control animals in size. There were no fundamental differences in protein:carbohydrate consumption patterns or strategies among experimental diets and larval instars. However, inadequate dietary water appeared to be more important for early than late instar larvae. Larvae on all artificial diets showed increasing fat content throughout all stages, including wandering and metamorphosis. Compensatory feeding among low-water and low-protein larvae was correlated with significantly higher fat content in larvae, pupae and adults, whereas low-sucrose animals were substantially leaner than those on the control diet. These differences may have strong effects on adult physiology, reproduction, and foraging patterns.     

Changes in lipid and fatty acid composition of eggs during development of the beet armyworm, Spodoptera exigua

The dry weight of Spodoptera exigua eggs decreased by 15·9 μg/egg or 64% of initial weight during embryogenesis and development of pharate first instar larvae. Lipid depletion accounts for 36% of this total dry weight loss and this occurs at an essentially constant rate throughout development. This marks S. exigua as an exception since most insects utilize lipids more rapidly during later developmental stages. Lipid depletion is due primarily to triglyceride catabolism, although phospholipids also decrease significantly.Fatty acid composition remains stable during development. In triglycerides, 18:1 is most common followed by 16:0 and 18:2; in the phospholipids, the order of abundance is 18:1, 18:2, and 16:0. Egg fatty acids differ from dietary fatty acids: 16:1 comprises 7% of triglyceride fatty acids although it is not present in the larval media; 18:1 predominates in the egg whereas 18:2 is most abundant in the diet.     

A possible rôle for microsomal oxidases in metamorphosis and reproduction in the housefly

Housefly larvae and adults of the Isolan-B strain, insecticide resistant because of high microsomal oxidase activity, and the susceptible WHO Standard Reference (SR) strain, were reared on diets containing phenobarbital or piperonyl butoxide. Periodically throughout the treatments groups of insects were assayed for microsomal aldrin epoxidase activity. This activity was compared with such growth and development parameters as pupation, adult emergence, and reproduction in similar groups of insects. At diet levels of 0·01 to 0·50 per cent both chemicals caused large increases, as much as elevenfold in microsomal oxidase activity in third instar larvae and 15 to 100 per cent inhibition of pupation and emergence. In the adult diet at 1 per cent, both compounds caused at least 50 per cent decrease in egg production. Phenobarbital enhanced the enzyme system in both larval and adult stages of both strains but piperonyl butoxide, while enhancing enzyme activity in larvae of the Isolan-B strain, was an inhibitor in the adult stage. The results are interpreted as an indication of a direct connexion between microsomal oxidase activity and the action of hormones in the housefly, probably through regulation of hormone titre by these enzymes.     

The essential dietary fatty acid requirement of the tufted apple budmoth, Platynota idaeusalis

The tufted apple budmoth, Platynota idaeusalis (Walker), was reared non-axenically for two successive generations on a casein-based semisynthetic diet. The qualitative essential fatty acid requirement for growth, development and normal pupal-adult ecdysis was studied using the non-axenic casein-based semisynthetic diets with and without various 99% pure fatty acids. Linoleic or linolenic acids caused accelerated larval development; linoleic, linolenic and arachidonic acids showed similar activity in body weight gain and survival to pupal-adult ecdysis. Linoleic or linolenic acids were active in alleviating wing deformities; arachidonic acid was partially active in alleviating wing deformities at the one dietary concentration evaluated. Activity of arachidonic acid as an essential fatty acid for P. idaeusalis is unique among insects, except for mosquitoes. The essential fatty acid deficiency syndrome of the adult, resulting from the larvae feeding on fat-deficient diets, was greatly reduced when larvae were fed on a diet adequate in essential fatty acid during either their early or late development.     

Self-selection of an optimum diet from a mixture of wheat fractions by the larvae of Tribolium confusum

Tribolium confusum was reared from the beginning of the first instar to the pupa on wheat bran, endosperm or germ or on a 1 : 1 : 1 mixture of these three fractions. The food intake of the larvae reared on the mixture included more or less similar proportions of the three wheat fractions, averaging 1·5 per cent bran, 17·1 per cent endosperm and 81·4 per cent germ. Past work on the nutrition of stored-product insects has involved the assumption that the insects cannot feed selectively from a finely powdered diet. Our findings show that this assumption may not be warranted. The mixed diet supported growth better than any one of the pure fractions or any wheat diet previously tested by us. It was superior to germ, the best of the pure fractions, because it was more digestible and because the digested portion was more efficiently utilized for growth. Endosperm, the poorest diet of the fractions, was highly digestible but was not efficiently utilized. Bran, better than endosperm but inferior to germ, was relatively indigestible although the digested portion was efficiently utilized. Germ was both digestible and efficiently utilized.     

A granulosis virus,possible biological agent for control of Adoxophyes orana (Lepidoptera: Tortricidae) in apple orchards: I. Mass production

Young larvae of Adoxophyes orana (apple race) were infected with a granulosis virus by feeding them artificial diet containing the virus suspension. The susceptibility decreased remarkably after the fourth to fifth instar. Infected larvae survived the end of the fifth (last) instar, even when they were infected at an early developmental stage. Inoculation of egg masses by dipping them into a virus suspension gave a high percentage of infection when larvae were reared en masse.     

Effects of azadirachtin on the nutrition and development of the tobacco budworm,Heliothis virescens (Fabr.) (Noctuidae)

The plant chemical azadirachtin was administered, either in artificial diet or by oral injection, to fifth instar larvae of the tobacco budworm, Heliothis virescens (Fabr.). At a dietary concentration of 0.03125 ppm, azadirachtin significantly reduced the amount of diet consumed and the weight gained by the larvae. Higher dietary concentrations (0.25 and 0.5 ppm) were necessary to reduce the efficiency of larval conversion of digested and ingested food, respectively. However, the approximate digestibility increased at the dietary concentration of 0.25 ppm.Orally injected azadirachtin (0.25 and 0.5 μg) delayed moulting to the pupal stage, produced defective pupae or adults, and inhibited development to the adult stage. Higher doses (5.0 and 10.0 μg) reduced the pre-pupal weight loss normally associated with pupation, and completely inhibited pupation. At the critical dose of 1.0 μg (the minimal dose that disrupted development to the pupal stage), azadirachtin had less of an effect on older than on younger larvae. Larvae injected on the first day of the fifth instar failed to pupate, whereas approx 40% of those injected on subsequent days pupated.The results suggest that azadirachtin affects H. virescens in a manner similar to other tested species of insects. The significance of these results, especially regarding hormonal events in the insects, is discussed.