Absorption of sugars and amino acids by the epidermis of Aphidius ervi larvae

Aphidius ervi Haliday (Hymenoptera, Braconidae) is an endophagous parasitoid of several aphid species of economic importance, widely used in biological control. The definition of a suitable artificial diet for in vitro mass production of this parasitoid is still an unresolved issue that, to be properly addressed, requires a deeper understanding both of its nutritional needs and of the functional properties of the larval epithelia involved in nutrient absorption. The experimental evidence presented in this paper unequivocally demonstrates that the uptake of sugars and amino acids takes place through the body surface of the larval stages of A. ervi. These nutrients are efficiently absorbed by the larval epidermis, but the transport rate progressively declines over time. The epidermis exhibits a cross-reactivity to antibodies raised against the mammalian facilitative glucose transporter GLUT2 and the sodium cotransporter SGLT1. The analysis of sugar transport sensitivity to specific inhibitors indicates the involvement of GLUT2-like transporters, while a role for SGLT1-like transporters is not supported. The peculiar pathways of nutrient absorption in A. ervi larvae further corroborate the general idea that the pre-imaginal stages of endophagous koinobiont Hymenoptera, like Metazoan parasites, show a high degree of physiological integration with their hosts.     

Unexpected similarity of intestinal sugar absorption by SGLT1 and apical GLUT2 in an insect (Aphidius ervi, Hymenoptera) and mammals

Sugars are critical substrates for insect metabolism, but little is known about the transporters and epithelial routes that ensure their constant supply from dietary resources. We have characterized glucose and fructose uptakes across the apical and basolateral membranes of the isolated larval midgut of the aphid parasitoid Aphidius ervi. The uptake of radiolabeled glucose at the basal side of the epithelium was almost suppressed by 200 microM cytochalasin B, uninhibited by phlorizin, and showed the following decreasing rank of specificity for the tested substrates: glucose > glucosamine > fructose, with no recognition of galactose. These functional properties well agree with the expression of GLUT2-like transporters in this membrane. When the apical surface of the epithelium was also exposed to the labeled medium, a cation-dependent glucose uptake, inhibited by 10 microM phlorizin and by an excess of galactose, was detected suggesting the presence in the apical membrane of a cation-dependent cotransporter. Radiolabeled fructose uptakes were only partially inhibited by cytochalasin B. SGLT1-like and GLUT5-like transporters were detected in the apical membranes of the epithelial cell by immunocytochemical experiments. These results, along with the presence of GLUT2-like transporters both in the apical and basolateral cell membranes of the midgut, as we recently demonstrated, allow us to conclude that the model for sugar transepithelial transport in A. ervi midgut appears to be unexpectedly similar to that recently proposed for sugar intestinal absorption in mammals.     

Functional analysis of a fatty acid binding protein produced by Aphidius ervi teratocytes

Aphidius ervi (Hymenoptera, Braconidae) is an endophagous parasitoid of various aphid species, including Acyrthosiphon pisum (Homoptera, Aphididae), the model host used in the present study. Parasitized hosts show a marked increase of their nutritional suitability for the developing parasitoid larvae. This alteration of the biochemical and metabolic profile is due to a castration process mediated by the combined action of the venom, injected at the oviposition, and of the teratocytes, cells deriving from the dissociation of the embryonic membrane. Teratocytes produce and release in the host haemocoel two parasitism-specific proteins, which are of crucial importance for the development of their sister larvae. One of the proteins is a fatty acid binding protein (Ae-FABP), which shows a high affinity for C14-C18 saturated fatty acids (FAs) and for oleic and arachidonic acids. To better define the possible nutritional role of this protein, we have studied its immunolocalization profile in vivo and the impact on FA uptake by the epidermal and midgut epithelia of A. ervi larvae. During the exponential growth of A. ervi larvae, Ae-FABP is distributed around discrete lipid particles, which are abundantly present in the haemocoel of parasitized host aphids and in the midgut lumen of parasitoid larvae. Moreover, a strong immunodetection signal is evident on the surface of the two larval epithelia involved in nutrient absorption: the parasitoid midgut epithelium and the external epidermal layer. These two epithelia can effectively absorb radiolabelled myristic acid, but the FA transport rates are not affected by the presence in the medium of Ae-FABP. The protein appears to act essentially as a vector in the host haemolymph, transferring FAs from the digestion sites of host lipids to the growing parasitoid larvae. These data indicate that the proteins produced by A. ervi teratocytes may play complementary roles in the nutritional exploitation of the host.     

Larval anatomy and structure of absorbing epithelia in the aphid parasitoid Aphidius ervi Haliday (Hymenoptera, Braconidae)

The present work describes Aphidius ervi Haliday (Hymenoptera, Braconidae) larval anatomy and development, focusing on time-related changes of body structure and cell ultrastructure, especially of the epithelial layers involved in nutrient absorption. Newly hatched 1st instar larvae of A. ervi are characterised by gut absence and a compact cluster of cells makes up their body. As the parasitoid larva develops, the central undifferentiated cell mass becomes hollowed out, leading to the formation of gut anlage. This suggests that absorption of nutrients at that stage may take place through the body surface, as more directly demonstrated by the occurrence on the epidermis of proteins associated with transepithelial transport, such as Na(+)/K(+)-ATPase and alkaline phosphatase (ALP). Second instar larvae show the presence of the gut with a well-differentiated brush border and a peritrophic membrane. Gut cells are filled by masses of glycogen granules and lipid droplets. The tracheal system starts to be visible. The haemocoel becomes evident in late 2nd instar, and contains large silk glands. Mature 3rd instar larvae are typically hymenopteriform. The midgut accounts for most of the body volume and is actively involved in nutrient absorption, as indicated by the well developed brush border and by the presence of Na(+)/K(+)-ATPase and ALP on the basolateral and luminal membrane respectively. At this stage, large lipid droplets have gradually replaced the cellular glycogen stores in the midgut cells. The tracheae are completely differentiated, but their internal lumen still contains fibrillar material, suggesting that they are not functional as long as host fluids bath the parasitoid larva. In late 3rd instar larvae, silk glands, structurally similar to Malpighian tubules, show a very intense vesicular traffic toward the internal lumen, which, eventually, results in being filled by secretion products, suggesting the possible recycling of metabolic waste products during mummy formation.     

Leucine transport by the larval midgut of the parasitoid Aphidius ervi (Hymenoptera)

The larval midgut of the hymenopteran parasitoid Aphidius ervi accomplishes a large transport of nutrients from the lumen to the haemocoel, providing most of the organic molecules necessary for rapid insect development. l-amino acids in general, and leucine in particular, are efficiently accumulated in the larval body. We show here that the intact midgut of early 3rd instar larvae incubated in vitro can take up [³H]l-leucine from the basolateral side of the epithelium by transporters insensitive to the presence of monovalent cations. When the midgut is opened and the apical membrane of the absorbing epithelial cells is exposed to the medium containing radiolabelled leucine, a sodium-dependent uptake of the amino acid becomes apparent, disclosing the presence of a symport mechanism. Inhibition experiments of leucine uptake by a 100-fold excess of different amino acids, selected according to the properties of their side chain, revealed that this apical sodium-dependent mechanism is a broad spectrum transport system with a specialization for the absorption of aliphatic amino acids, that can also transfer glutamine and proline, but not phenylalanine, lysine and arginine. Altogether the experimental results obtained with intact- and open-gut preparations suggest that leucine transport across the basolateral membrane is mediated by both an uniporter and an obligatory amino acid exchange mechanism.     

Modification of the nutritional parameters and of midgut biochemical and absorptive functions induced by the IGR fenoxycarb in Bombyx mori larvae

Fifth instar larvae of B. Mori were topically or orally treated with increasing amounts of the Insect Growth Regulator (IGR) fenoxycarb in a single application, in order to determine its effects on the nutritional parameters, the midgut functional activities and the growth of the silk glands. The IGR affected in a dose-dependent manner the progress of the life cycle of the insect, causing a delay or inhibition of spinning, alteration of the feeding behaviour, decrease of the nutritional parameters, impairment of the growth of the silk glands, and an increased mortality during larval-pupal transformation. Measurement of leucine uptake into midgut brush border membrane vesicles and midgut histochemistry revealed a reduced absorption of leucine by the midgut and a large alteration of a number of midgut enzyme activities as a result of treatments with a high dose of fenoxycarb (2.5 μg). Treatments with a dose of 2.5 femto g/larva caused an increase in leucine uptake by the midgut, an increased weight of the cocoon shell, and a modification of some midgut enzyme activities. The lepidopteran midgut appears to be a larval organ that responds promptly to the exposure to fenoxycarb. The epithelial columnar cells modify their absorptive functions, at least with regard to amino acid uptake, as well as their metabolic activity, with a modification of the oxidative status of the cells that is detectable with a single dose of the chemical as low as few fg/larva. Arch. Insect Biochem. Physiol. 39:18–35, 1998. © 1998 Wiley-Liss, Inc.     

Influence of the parasitoid Chelonus inanitus and its polydnavirus on host nutritional physiology and implications for parasitoid development

Chelonus inanitus is a solitary egg-larval endoparasitoid, which feeds on host haemolymph during its internal phase. Parasitization induces in the host Spodoptera littoralis a precocious onset of metamorphosis and a developmental arrest in the prepupal stage. At this stage the parasitoid larva emerges from the host and consumes it. We show here that parasitization and the co-injected polydnaviruses affect the nutritional physiology of the host mainly in the last larval instar. Polydnaviruses cause a reduced uptake of food and an increase in the concentration of free sugars in the haemolymph and of glycogen in whole body. The parasitoid larva, along with polydnaviruses, causes a reduction of proteins in the host's plasma and an accumulation of lipids in whole body. Dilution of host haemolymph led to a reduced concentration of lipid in parasitoid larvae and a reduced survival rate. Thus, a sufficient concentration of nutrients in the host's haemolymph appears to be crucial for successful parasitoid development. Altogether, the data show that the parasitoid and the polydnavirus differentially influence host nutritional physiology and that the accumulated lipids and glycogen are taken up by the parasitoid in its haematophagous stage as well as through the subsequent external host feeding.     

Modulation of leucine absorption in the larval midgut of Bombyx mori (Lepidoptera, Bombycidae).

In the larval midgut of Bombyx mori a K(+)-dependent transporter for leucine and amino acids with a hydrophobic side chain is responsible for the absorption of most essential amino acids. We investigated if a modulation of its activity occurred as a result of starvation or after hormonal treatments. We measured amino acid uptake in brush border membrane vesicles (BBMV) purified from the anterior-middle (AM) and posterior (P) regions of the midgut in fifth instar larvae. Silkworms were either starved or topically treated with low dosages of fenoxycarb, a molecule often used as a juvenile hormone mimic. The maximal uptake value of K(+)-driven leucine transport was increased in BBMV of AM- and P-midgut regions of starved larvae. The initial uptake rates of serine and glutamine, two amino acids transported by the same cotransporter as leucine, were also increased. Leucine kinetics proved that V(max) was the kinetic parameter modified by starvation in both midgut regions. Topical applications of fenoxycarb at a dose of 2.5 fg/larva immediately after the fourth ecdysis, induced an increase of leucine initial uptake rates and of intravesicular accumulation of leucine in both AM- and P-BBMV. Kinetic analysis of leucine uptake indicated again that V(max) was increased in BBMV from both midgut regions in treated larvae.     

幼虫密度对草地螟食物利用率及消化酶活性的影响

为了阐明草地螟Loxostege sticticalis大发生种群幼虫取食行为特征, 在室内条件下（温度22±1℃, 相对湿度70%）对不同幼虫密度［1, 10, 30头/瓶（650 mL）］饲养草地螟幼虫的食物利用率及消化酶活性进行了研究。结果表明： 幼虫中等（或高）密度对草地螟幼虫相对中肠重量、 相对取食量、 粪便干重、 食物利用率和近似消化率及总蛋白酶和亮氨酸氨肽酶活性影响显著。幼虫相对中肠重量以10头/瓶的幼虫密度最大, 1头/瓶的幼虫密度最小。随着幼虫密度的增加, 幼虫相对取食量和粪便干重增加, 而虫体干重减轻, 幼虫食物利用率降低。幼虫密度30头/瓶的幼虫相对取食量和粪便干重显著高于1和10头/瓶的, 而30头/瓶的幼虫食物利用率显著低于1头/瓶的。幼虫近似消化率随幼虫密度的逐渐增加而显著降低。幼虫密度10头/瓶的幼虫总蛋白酶和亮氨酸氨肽酶的活性显著高于1和30头/瓶的, 而淀粉酶的活性受幼虫密度影响不显著。随幼虫密度的增加, 幼虫相对中肠重量与总蛋白酶和亮氨酸氨肽酶活性变化趋势较为一致, 消化酶活性的变化可能与相对中肠重量大小有关。因此, 幼虫密度是影响草地螟幼虫取食行为的重要因子, 这些结果为阐明草地螟大发生种群与一般种群的为害特征提供了重要理论依据。     

Host hemolymph proteins and protein digestion in larval Habrobracon hebetor (Hymenoptera: braconidae)

Host plasma proteins and protein digestion in larval parasitoids were studied during trophic interactions of the ectoparasitoid Habrobracon hebetor Say (Hymenoptera: Braconidae), with a host, larvae of the Indianmeal moth, Plodia interpunctella Hübner (Lepidoptera: Pyralidae). We could detect no apparent differences in host hemolymph protein patterns up to 72 h after paralysation and/or parasitization by H. hebetor. A 190 kDa putative apolipophorin I present in host hemolymph could not be detected in the midguts of feeding H. hebetor larvae indicating that it is rapidly digested. The major 60 kDa storage proteins (putative hexamerins) in host hemolymph were detected in the parasitoid midgut and were completely digested 24 h after cessation of feeding and the beginning of cocoon formation. Host hemolymph had a pH of about 6.4. The pH optima of the midgut proteinases in the larval parasitoid were in the alkaline region, but midgut fluid in feeding parasitoid larvae was about pH 6. 8. Based on enzyme activity against selected artificial proteinase substrates including azocasein, N-alpha-benzoyl-L-Arg p-nitroanilide (BApNA), succinyl-Ala-Ala-Pro-Phe p-nitroanilide (SAAPFpNA), succinyl-Ala-Ala-Pro-Leu p-nitroanilide (SAAPLpNA), and inhibition by selected proteinase inhibitors, serine proteinases appear to be the predominant class of enzymes involved in protein digestion in the midguts of H. hebetor. There is also an active aminopeptidase (LpNA) associated with the microsomal fraction of midgut preparations. There was no evidence for preoral digestion or ingestion of proteinases from host hemolymph by the parasitoid larva. There was a very active BApNAase in the soluble fraction of midgut extracts. This activity increased on a per midgut basis up to 24 h after the beginning of cocoon formation but decreased rapidly by 48 h. Two major (P1 and P3) and several minor proteinases were detected in midgut extracts of H. hebetor analysed with gelatin zymograms. The apparent molecular mass of P1 varied from 95 to 49 kDa depending on protein loading. P3 had an apparent molecular mass of 39 kDa that was independent of protein loading. In summary, electrophoretic evidence indicates that host hemolymph protein patterns do not change significantly for at least 72 h after paralysation by H. hebetor. The role, if any, of envenomization in preventing breakdown of hemolymph proteins during this time remains to be determined. Because the predominant host hemolymph proteins, a putative apolipophorin I and the putative hexamerins, are readily digested by the serine proteinases present in the midguts of this parasitoid larva, these or similar proteins would provide an easily digested source of dietary amino acids that could be used for development of artificial diets for this beneficial insect.     

菜蛾盘绒茧蜂对寄主小菜蛾幼虫营养的调节和利用

寄主小菜蛾Plutella xylostella被内寄生蜂菜蛾盘绒茧蜂Cotesia plutellae寄生后,其取食、发育及营养代谢在各种寄生因子的作用下伴随幼蜂的发育而发生很大的变化,畸形细胞作为调节因子之一也发挥了重要的作用。本实验通过比较被寄生和未被寄生小菜蛾血淋巴蛋白浓度以及两种血淋巴对菜蛾盘绒茧蜂幼蜂进行体外培养的培养液的蛋白浓度,发现被寄生小菜蛾血淋巴比未被寄生小菜蛾血淋巴的蛋白浓度略低但差异不显著,而未被寄生小菜蛾血淋巴幼蜂培养液的蛋白浓度显著低于被寄生小菜蛾血淋巴幼蜂培养液的蛋白浓度,证明畸形细胞的蛋白质分泌功能。被寄生后期, 小菜蛾体重明显大于未被寄生的小菜蛾体重,而脂肪体重量相比正好相反;通过显微染色观察,在小菜蛾念珠状脂肪体表面粘附有畸形细胞,对脂肪体进行分解破坏而使其成颗粒状; 蛋白含量和脂滴浓度测定也表明,脂肪体的可溶性蛋白含量和脂滴浓度也迅速降低,同比低于未被寄生小菜蛾。而与此同时,幼蜂正处在快速生长阶段,中肠酯酶的活性逐步上升,幼蜂得以快速消化吸收小菜蛾体内的营养直到完成幼虫发育,整个幼蜂的脂滴浓度也达到了最大值。因此寄生后期,推测在畸形细胞的协助下,幼蜂吸收了寄主小菜蛾体内的营养为自身生长发育所用。     

Stage-dependent strategies of host invasion in the egg-larval parasitoid Chelonus inanitus

Chelonus inanitus (Braconidae) is a solitary egg-larval parasitoid which lays its eggs into eggs of Spodoptera littoralis (Noctuidae); the parasitoid larva then develops in the haemocoel of the host larva. Host embryonic development lasts approx. 3.5 days while parasitoid embryonic development lasts approx. 16 h. All stages of host eggs can be successfully parasitized, and we show here that either the parasitoid larva or the wasp assures that the larva eventually is located in the host's haemocoel. (1) When freshly laid eggs, up to almost 1-day-old, are parasitized, the parasitoid hatches while still in the yolk and enters the host either after waiting or immediately through the dorsal opening. (2) When 1-2-day-old eggs are parasitized, the host embryo has accomplished final dorsal closure and is covered by an embryonic cuticle when the parasitoid hatches; in this case the parasitoid larva bores with its moving abdominal tip into the host. (3) When 2.5-3.5-day-old eggs are parasitized, the wasp oviposits directly into the haemocoel of the host embryo; from day 2 to 2.5 the embryo is still very small and the wasps, after probing, often restrain from oviposition for a few hours.     

Role of specific activators of intestinal amino acid transport in Bombyx mori larval growth and nutrition.

Nutrient absorption and its modulation are critical for animal growth. In this paper, we demonstrate that leucine methyl ester (Leu-OMe) can greatly increase the activity of the transport system responsible for the absorption of most essential amino acids in the larval midgut of the silkworm Bombyx mori. We investigated leucine uptake activation by Leu-OMe in brush border membrane vesicles and in the apical membrane of epithelial cells in the midgut incubated in vitro. Moreover, the addition of this strong activator of amino acid absorption to diet significantly affected larval growth. Silkworms fed on artificial diet supplemented with Leu-OMe reached maximum body weight 12-18 h before control larvae, and produced cocoon shells up to 20% heavier than those of controls. The activation of amino acid absorption plays an essential role in larval development so that larval growth and cocoon production similar to controls reared on an artificial diet with 25% of dry mulberry leaf powder were observed in silkworms fed on an artificial diet with only 5% of mulberry powder. Arch.     

Plant virus and parasitoid interactions in a shared insect vector/host

Interactions between barley yellow dwarf luteovirus (BYDV) and the aphid parasitoid, Aphidius ervi Haliday (Hymenoptera: Aphidiidae), were investigated while sharing the vector/host, Sitobion avenae (F.) (Homoptera: Aphididae). Aphids, which were parasitized during their second larval stadium, had access to virus-infected plants before, immediately after, or several days after parasitoid attack. The larval development of A. ervi in S. avenae was significantly delayed when virus acquisition took place before or shortly after the parasitoid had hatched, but not when the parasitoid was at the second larval stage during virus acquisition. Similarly, the presence of BYDV led to a significantly higher aphid mortality when they acquired virus up to and including the time that A. ervi was at the first larval stage. Adult female parasitoids deposited fewer eggs in viruliferous aphids. Virus transmission was not reduced by parasitization, and in some experiments aphids which were subjected to parasitoid attack transmitted BYDV more efficiently than unattacked insects.     

Larval competition between Aphidius ervi and Praon volucre (Hymenoptera: Braconidae: Aphidiinae) in Macrosiphum euphorbiae (Hemiptera: Aphididae)

Interspecific competition between parasitoid larvae may influence the size, structure, and stability of the population, leading to a reduction in total parasitism and thus restricting the pest control. Aphidius ervi (Haliday) and Praon volucre (Haliday) are endoparasitoids that possess a wide host range and present considerable potential for the biological control of the aphid Macrosiphum euphorbiae (Thomas). The larval competition between A. ervi and P. volucre, and the possible intrinsic competitive superiority of one of the parasitoids in M. euphorbiae, have been studied. In single parasitism experiments, mated parasitoid females (n=10) were maintained individually in contact with M. euphorbiae hosts (n=30) inside petri dishes containing lettuce leaf discs and maintained in environmental chamber at 22 ± 1°C, 70 ± 10% RH, and 12-h photophase. The multiple parasitism experiments consisted of exposing single parasitized aphids (n=120) to the second parasitoid species. Two oviposition events were performed with a 4-h interval between them, namely the following: sequence A (oviposition by A. ervi, followed by P. volucre) and sequence B (oviposition by P. volucre, followed by A. ervi). Oviposition sequence A generated 24 A. ervi and 55 P. volucre adults, whereas oviposition sequence B generated 23 and 49 adults. P. volucre is an intrinsically superior competitor compared with A. ervi, and the use of the two species simultaneously may result in competitive exclusion and influence the stability of the parasitoid population.     

Effects of rearing density on larval growth and activity of digestive enzymes in Lymantria dispar L. (Lepidoptera: Lymantriidae)

Density dependent responses of 4th, 5th and 6th instar gypsy moth larvae were studied at the level of larval mass, midgut loading and activities of three digestive enzymes (alpha-amylase, trypsin and leucine aminopeptidase). High density significantly reduced larval mass while midgut loading (expressed as relative midgut mass) did not change except in the 5th instar where it was increased at high density. Specific amylase and leucine aminopeptidase activities were not affected by crowding. Specific trypsin activity was on average higher in crowded than in isolated larvae. High density also affected the correlations between midgut protein content and activities of two proteolytic enzymes suggesting differences in regulatory mechanisms of insect digestion. The importance of these changes for survival under stressful conditions is discussed.     

Rhythmic contractile movements of the larval midgut of the silkworm, Bombyx mori

The raidgut of intact larvae of Bombyx mori exhibited active contractile movements. Contraction waves were generated rhythmically at several regions of the midgut. The waves passed in both oral and aboral directions from their sites of origin. Midgut movements were depressed during moulting. The midgut continued to move normally when tetrodotoxin was injected into the larval haemocoel at doses sufficient to paralyze somatic muscle. Ligation of larvae paralyzed with tetrodotoxin behind the second or third body segment resulted in the abolition of the contraction waves in the midgut portion anterior to the ligature. A ligature applied behind other body segments did not hamper midgut motor activity irrespective of whether or not an abdominal ganglion had been extirpated. The frequency of contraction and the rate of food transport in the midgut were increased when larvae were administered serotonin or when their body temperature was raised.     

A novel regulatory mechanism for amino acid absorption in lepidopteran larval midgut

A number of methyl and ethyl esters of naturally occurring amino acids exert a potent stimulatory effect on the cotransport system responsible for the absorption of most essential amino acids along the midgut of the silkworm Bombyx mori. L-Leucine methyl ester (Leu-OMe), one of the most effective activators, induces a large increase of the initial rate of leucine uptake in midgut brush border membrane vesicles (BBMV) from the anterior-middle (AM) region, and a small effect in BBMV from the posterior (P) region. Nonetheless, the methyl ester causes in both regions a relevant K(+)-, Deltapsi- and pH-independent increase of the intravesicular accumulation of the amino acid. The activation by Leu-OMe proves that amino acid absorption can be modulated all along the B. mori larval midgut and that the AM region, where the ability to transport and concentrate the substrate is very low, is more susceptible than the P region. Leucine uptake in AM-BMMV can be activated by amino acid methyl esters with definite structural requisites, with the following order of potency: L-leucine>L-phenylglycine>L-methionine>L-phenylalanine>L-norleucinez.Gt;L-isoleucine. The activation is stereospecific and occurs also with some ethyl esters (e.g. leucine and phenylalanine). No activation was observed with esters of amino acids with short hydrophobic or polar side-chains. The activation mechanism here described plays a fundamental role in larval growth since silkworms reared on artificial diets supplemented with leucine or methionine methyl esters reach maximum body weight 12-18 h before control larvae and spin cocoons with a larger shell weight. This novel regulatory mechanism of an amino acid transport protein appears to be widespread among lepidopteran larvae.     

Uptake of bovine IgG by first instars of the common cattle grub, Hypoderma lineatum

The midgut of first instar Hypoderma lineatum (De Vill.) (Diptera: Oestridae) was examined for the presence of bovine immunoglobulin using immuno-gold staining in conjunction with image analysis. Newly hatched larvae, unexposed to fluids or tissues of the host, showed little or no deposit of colloidal gold particles. Midgut lumen, microvillar border, epithelial cell bodies and basement membrane of first instars, 6-7 months of age, recovered from the host oesophagus, all showed colloidal gold staining. This indicates that immuno-reactive portions of bovine immunoglobulin survive exposure to extracorporeal larval enzymes, as well as transit of the midgut, and pass through epithelial cells to enter the larval haemocoel.     

Interactions between an entomopathogenic microsporidium and the endoparasitoid Glyptapanteles liparidis within their host, the gypsy moth larva

Interactions in the host-parasitoid-pathogen system, Lymantria dispar L. (Lep., Lymantriidae)-Glyptapanteles liparidis (Bouché) (Hym., Braconidae)-Vairimorpha sp. (Protista, Microspora), were investigated. Host selection experiments revealed that G. liparidis females did not discriminate between infected and uninfected host larvae for oviposition. Transmission of the microsporidium from infected to uninfected hosts by stinging female wasps could not be ascertained. Females that developed in infected L. dispar larvae did not transmit the pathogen via oviposition. Vairimorpha infection of the host negatively affected the performance of the braconid, when inoculation took place either before or after parasitization. Microsporidiosis of the host caused delayed development, reduced pupation and adult eclosion, reduction in size and weight, and reduction of adult longevity of G. liparidis. Parasitoids themselves were not systemically infected by Vairimorpha sp., but braconid larvae did ingest microsporidian spores at the end of their endoparasitic development and accumulated the undigested and ungerminated spores in the blind midgut. Negative effects of host infection on parasitoid larvae were detectable from the beginning of parasitoid larval development. Lethal time was reduced when L. dispar larvae were infected and parasitized, often at the expense of the parasitoid when G. liparidis were unable to complete endoparasitic development before the host died. Intensity of infection, measured as number of spores produced per milligram fresh weight of L. dispar larva, was slightly higher in parasitized and infected hosts than in unparasitized and infected hosts.