A new cell line from larval fat bodies of the bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae)

Summary A new cell line, designated IOZCAS-Ha-I, was initiated from the fat body of larvae of Helicoverpa armigera (Lepidoptera: Noctuidae) in TNM-FH medium containing 10% fetal bovine serum. Spherical cells were predominant among the various cell types. The cell line showed a typical lepidopteran chromosome pattern ranging from 58 to 239 chromosomes in the majority of the cells, it was confirmed to have originated from the H. armigera by the DNA amplification-fingerprinting polymerase chain reaction (DAF-PCR) technique. The new cell line was only slightly susceptible to the multiple nucleocapsid nuclear polyhedrosis viruses (NPV) from H. armigera.     

Expression of the cry1EC gene in castor (Ricinus communis L.) confers field resistance to tobacco caterpillar (Spodoptera litura Fabr) and castor semilooper (Achoea janata L.)

Castor (cv. DCS-9) has been transformed through Agrobacterium-mediated and particle gun bombardment methods using appropriate vectors containing the Bt chimeric gene cry1EC driven by enhanced 35S promoter. About 81 and 12 putative transformants were regenerated following selection on hygromycin and kanamycin, respectively. Southern analysis of DNA extracted from T₀ plants confirmed integration of the introduced gene in castor genome. The integration and inheritance of the introduced genes was demonstrated up to T₄ generation by PCR and Southern analysis. Southern analysis of two events having single and two copies showed the same pattern of integration in the subsequent generations. Insect feeding experiments conducted in the laboratory by releasing neonate larvae of castor semilooper and S. litura on leaf tissues excised from transgenic and control plants showed varying degrees of larval mortality and slow growth in larvae fed on transgenic leaf tissue. Field bioassays against Spodoptera litura and castor semilooper conducted for eight events in T₁–T₄ generations under net confinement were more informative and events conferring resistance to the two major defoliators were identified.     

Galleria mellonella larvae fat body disruption (Lepidoptera: Pyralidae) caused by the venom of Habrobracon brevicornis (Hymenoptera: Braconidae)

The ability of Habrobracon brevicornis venom to elevate the nutritional suitability of a host by affecting the host larvae fat body condition was studied. To understand whether H. brevicornis crude venom impacts the host biochemical profile, the concentrations of total lipids and main sugars in the host larvae lymph were analyzed. All measurements were carried out during the first 3 days after envenomation. A significant increase in the lipid level was fixed only on the second day after envenomation. A significant increase in the total trehalose count was detected during all 3 days, while a significant increase in glucose concentration was noted only on the first day. Well‐observed disruptions were fixed in thin and semithin sections of the G. mellonella larval fat body starting from the second day after envenomation. Significant increases in both phospholipase A₂ and C enzyme activity as well as acid proteases were detected in the wax moth fat body after envenomation during all experimental times. At the same time, imbalances in the antioxidant system, including changes in the activities of superoxide dismutase, peroxidases, catalase, and glutathione‐S‐transferase, were detected. The reliable increase in the expression of the gene encoding Hsp70 was fixed both for 24 and 48 h after envenomation, while a reliable increase in the expression of the gene encoding inhibitor of apoptosis protein was detected only 24 h after wax moth larvae envenomation. Considering the absence of DNA fragmentation, the imbalance in the “ROS/antioxidants” system, and the increased activity of phospholipases and acid proteases in the fat body cells from envenomated wax moth larvae, we can hypothesize that the fat body disruption occurs in a necrotic manner. The results of the work expand the knowledge about the biochemical aspects of interaction between ectoparasitoids and their hosts     

Hormonal control of lipid synthesis in the fat body of the adult female tsetse fly, Glossina morsitans

Aqueous extracts of brain, thoracic ganglion or corpora cardiaca of female Glossina morsitans were shown to contain a substance which inhibited the synthesis of lipid from l[U-¹⁴C] leucine by fat cells incubated in vitro. The highest concentration of this substance was found in the corpora cardiaca; approximately 1 × 10^?6 gland pairs μl^?1 were required for maximum inhibition. At concentrations greater than 1 × 10^?4 gland pairs μl^?1 the lipid synthesis inhibiting factor (hereafter referred to as the LSIF) was inactivated by the presence of a substance which could be removed by gel filtration. The concentration of LSIF in the corpora cardiaca and midbrain varied throughout the reproductive cycle of the female. Net release of LSIF from the midbrain occurred between the 2nd and 7th day of the 9-day reproductive cycle. Net release from the corpora cardiaca began on day 5 and continued until the end of the interlarval period on day 9. Results are consistent with the hypothesis that LSIF is synthesised mainly in the medial neurosecretory cells of the midbrain whereas the corpora cardiaca are the site of storage and release into the haemolymph. LSIF was present in midbrain and corpora cardiaca extracts from male G. morsitans but at lower concentrations than in females. No variation in LSIF concentration could be correlated with the feeding cycle. LSIF activity was not detected in fresh haemolymph but was found at high concentration in boiled haemolymph, suggesting the presence of an inhibitor which was inactivated at high temperature. Preliminary investigations into the nature of LSIF have shown it to be inactivated by proteolytic enzymes and to be recoverable in a single peak from a Sephadex G15 column.Results support the view that LSIF is a peptide hormone which, in conjunction with an inhibitor, controls the lipid synthetic ability of the fat cells of the adult female tsetse fly throughout the reproductive cycle.     

Variations in body length, weight, fat content and survival inCoccinella septempunctata at different hibernation sites

Adults ofCoccinella septempunctata L. (Coleoptera; Coccinellidae) were sampled in four overwintering sites: one in England and three at different altitudes in the Czech Republic (350–1420 m above sea level) in autumn 1992 and spring 1993. Body length, dry weight and fat weight were measured. There were significant differences in body length between sampling sites in spring 1993, but not in autumn 1992. The average sex ratio (♀) in all samples was 1∶0.60 in 1992 and 1∶0.72 in 1993. Females were significantly larger and heavier, and contained more fat than males in both sampling periods. Dry weight and fat weight were positively correlated to body length at all sampling sites except at Ruzyne, in the Czech Republic. Adults hibernating at higher altitudes and away from their breeding and feeding habitats had significantly more fat than those hibernating at lower altitudes. Fat reserves were reduced by 30% during the hibernation at the top of the Krkonose mountain, in the Czech Republic, where temperatures were much lower, but more than half of the fat reserves were consumed during the overwintering period at the other sampling sites. The significance of body length and fat reserves in relation to adult hibernation sites and overwintering survival is discussed.     

On the importance of fat reserves for the little auk (Alle alle) chicks

Summary The pattern of fat deposition and inter-year variation in the amount of deposited fat, lean dry mass (LDM) and water were studied in the chicks of little auk (Alle alle), an arctic alcid, at Spitsbergen in the 1984, 1986 and 1987 breeding seasons. The lipid index (lipid mass/LDM) rose from 0.29 at hatching to about 0.70 at the age of maximum body mass attained in the nest, the highest yet recorded for seabirds except procellariiforms. Just before fledging the index dropped to 0.38. Fat mass in 1984 chicks and also fresh body mass of older 1984 chicks was significantly lower than in the other two seasons, but there were no statistical year-to-year differences in LDM's of chicks. The amount of fat in chicks was significantly correlated with certain weather parameters. Fasting capability (FC) of the chicks (the time that chicks could survive when using only their lipid stores) was calculated from fat mass and previous data on metabolic rates of chicks. The FC's of all 108 chicks collected in three seasons were greater than maximum interval between two consecutive feeds to chicks reported in the study colony. Also the estimated fat reserves of 99.7% of chicks weighed in 1984 were sufficient to support them for longer than the maximum recorded intervals between feeds. In relation to death by starvation, little auk chicks, at least up to the age of maximum body mass, have a substantial surplus of fat. This is contrary to the hypothesis that fat depots in seabird chicks assist survival during the periodic fasts resulting from unpredictable feeding conditions.     

Vitellogenin synthesis by the fat body of Porcellio dilatatus Brandt (Crustacea,Isopoda)

Using immunofluorescence, vitellogenin has been identified in the fat body of ovariectomized females of Porcellio dilatatus. Using immunoradiography of incubated fat body in leucine-¹⁴C-labeled medium, this organ has been demonstrated to be the synthesis site. Storage, but not synthesis, has also been proven in a particular kind of hepatopancreatic cell which might play a part in the resorption of vitellogenin excess.     

Variation in body composition of female big brown bats (Eptesicus fuscus) during lactation

Most small mammals support the nutritional requirements of milk production by increasing food intake. However, when nutrient intake is low, maternal body reserves may be mobilized to maintain adequate milk output. We examined patterns of body composition, including dry matter, fat, protein, and mineral content in big brown bats, Eptesicus fuscus, during lactation. Concentrations of fat and phosphorus were markedly lower in lactating mothers during week three of lactation than during the first two weeks, but these constituents rebounded to previous levels in the fourth and fifth week. Rapid recovery from fat depletion suggests that females are able to adjust to changes in demands for energy. The decrease in phosphorus during mid-lactation suggests bone demineralization, but an interspecific comparison of adult concentrations of minerals prevalent in bone suggests that mineral concentrations may never reach critically low levels in reproductively active females.     

Metabolism of juvenile hormone in cultures of ovaries and fat body in the cockroachperiplaneta Americana

Summary The time course of juvenile hormone (JH) metabolism is examined in cultures ofPeriplaneta americana fat body and ovaries in medium containingManduca sexta carrier protein or cockroach hemolymph. In the absence ofM. sexta carrier protein or cockroach hemolymph, both tissues extensively catabolize exogenous [³H]JH in the medium. Addition of the carrier protein or hemolymph to the culture system prevents the hydrolysis of the hormone in the medium. Within the tissues JH is degraded whether or not carrier protein or hemolymph is present which suggests that the protective role of these molecules is exclusively extracellular. Incubation of [³H]JH with medium preconditioned with tissue results in destruction of the hormone. This suggests that the fat body secretes esterases into the medium. In contrast, the ovarioles hydrolyze the hormone by means of cell-associated enzyme. The relationship of these phenomena to insect development is discussed. This work supported by NSF Grant PCM 76-02229 and University of Kansas Biomedical Sciences Grant RR-07037.     

Comparing the effects of the exotic cactus-feeding moth, Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae) and the native cactus-feeding moth, Melitara prodenialis (Walker) (Lepidoptera: Pyralidae) on two species of Florida Opuntia

This study examined the effects of the native cactus moth borer, Melitara prodenialis, and the invasive cactus moth borer, Cactoblastis cactorum, on two common cactus species, Opuntia stricta and O. humifusa at coastal and inland locations in central Florida. Opuntia stricta were present only at coastal sites and O. humifusa were present at coastal and inland sites. Throughout the duration of the study, coastal plants were subject to damage solely by C. cactorum and inland plants solely by M. prodenialis. Results showed marginally significantly higher numbers of eggsticks on O. stricta than O. humifusa and significantly higher numbers at coastal sites than at inland sites. There was also significantly higher moth damage on O. stricta than O. humifusa and at coastal sites than inland sites, but not significantly so. However, there was a higher level of plant mortality for O. humifusa than for O. stricta and a significantly higher level of cactus mortality at inland sites when compared to coastal sites. This increased mortality may be due to increased attack by true bugs, Chelinidea vittiger, and by Dactylopius sp., combined with attack by M. prodenialis. Inland plants also tended to be smaller than coastal plants and could be more susceptible to the combined effects of all insects. Further long-term research on coastal cactus survival when attacked and unattacked by Cactoblastis is necessary to fully determine the effects of this moth on Opuntia survival.     

Characterization of cockroach (Periplaneta americana) fat body phospholipase A(2) activity

A phospholipase has been identified in the fat body of the American cockroach, Periplaneta americana, which removes fatty acid from the sn-2 acyl position of an artificial substrate. The enzyme has been characterized using a crude preparation obtained by low-speed centrifugation of the homogenized tissue. With 1-hexadecanoyl-2-(1-pyrenedecanoyl)-sn-glycero-3-phosphocholine as the substrate, the K(m) has been estimated to be 1.17 microM and the v(max) 113.5 pmol/min/mg protein. The phospholipase has a pH optimum close to 7 and shows maximal activity at 50 degrees C. Activity of the phospholipase has been determined in cytosolic and plasma membrane fractions. The specific activity of the latter fraction is approximately twice that of the cytosol. The enzyme in both fractions is Ca(2+)-independent. Arch.     

Expression and localization of storage protein 1 (SP1) in differentiated fat body tissues of red hairy caterpillar, Amsacta albistriga Walker

The accumulation and utilization of storage proteins are prominent events linked to the metamorphosis of holometabolous insects. The female-specific storage protein 1 (SP1) is the major storage protein found in the hemolymph and fat body of female larvae of the groundnut pest, Amsacta albistriga. Here we show SP1 expression and localization in differentiated fat body tissues using biochemical and immunohistochemistry scrutiny. Comparison of A. albistriga SP1 with that of other species with respect to amino acid composition and N-terminal sequences show that SP1 is a methonine-rich protein and its identity was confirmed by means of immunoblot analysis. Northern blot studies revealed that the SP1 gene demonstrates stage- and tissue-specific expression in the peripheral fat body cells during the mid-larval period of fifth instar of A. albistriga. During the larval pupal transformation, SP1 are sequestered mainly by the perivisceral fat body tissues, until they serve the purpose of supplying amino acids for the production of egg yolk proteins. Further, electron microscopic studies using immunogold tracer techniques confirmed the localization of crystalline SP1 reserves, stored in the perivisceral fat body tissues. Hence, the peripheral fat body is responsible for biosynthesis of storage proteins, whereas the perivisceral fat body is a specialized storage organ.     

On the presence and localization of glycogen accumulations inside coronet cells of the saccus vasculosus of the dogfish (Scylliorhinus caniculus)

Summary In several coronet cells of the saccus vasculosus of Scylliorhinus large quantities of glycogen occur, as shown by light and electron microscopy. The significance of glycogen as an energy storage necessary for a transcellular ion transport process taking place in the coronet cells is discussed.The authors thank Dr. F.C.G. van de Veerdonk, W. F. Jansen and W. F. G. Flight for reading the manuscipt and for their critical remarks. They are also indebted to Mr. H. van Kooten and his staff for their valuable photographic assistance.     

Hormonal regulation of proline synthesis and glucose release in the fat body of the Colorado potato beetle, Leptinotarsa decemlineata

In the adult males of the Colorado potato beetle, Leptinotarsa decemlineata, corpora cardiaca extract injected in vivo gives rise to an increase in glucose and a decrease in alanine concentration of the haemolymph. The regulation of proline synthesis and glucose release in the fat body of the Colorado potato beetle was investigated further in vitro. Proline regulates its own synthesis by a feedback inhibition. Moreover, a factor present in extracts from the corpora cardiaca stimulates synthesis in the fat body in vitro. This effect was demonstrated with corpora cardiaca extracts from beetles, locusts and cokroaches. Also, synthetic adipokinetic hormone stimulates proline synthesis in the fat body of the Colorado beetle. In addition, a release of glucose from the fat body of the beetle could be evoked by the addition of locust and beetle corpora cardiaca extracts or synthetic adipokinetic hormone. The physiological significance of both effects is discussed.     

颈双缘姬蜂寄生引起小菜蛾蛹脂肪体形态和超微结构的变化

采用光学和电子显微镜观察了颈双缘姬蜂Diadromus collaris（膜翅目： 姬蜂科）寄生后小菜蛾Plutella xylostella（鳞翅目：菜蛾科）蛹脂肪体形态、超微结构和脂肪细胞的变化。结果表明： 被寄生72 h后小菜蛾蛹脂肪体结构松散,细胞游离,细胞膜破裂;细胞内营养物质开始被动地消耗;细胞器数量减少,细胞核内染色质状态发生变化。这些现象说明寄生对寄主的脂肪体结构及脂肪细胞产生了明显的影响,这有利于为幼蜂的发育提供营养。     

Effect of maternal fat reserves on the fatty acid composition of sardine (Sardina pilchardus) oocytes

We compared the fatty acid (FA) composition of the muscle and gonads of female Iberian sardines with hydrated oocytes collected during the 2002/03 spawning season off southern Portugal (November and February) and off western Portugal (February). Sardine condition and total FA concentration in the muscle decreased between the two sampling dates, while the gonadosomatic index was similar between samples. Total monounsaturated FA concentrations in sardine gonads were different for the three samples while saturated and polyunsaturated FA concentrations were similar. Significant linear relations were found between FA concentrations in female muscle and oocytes, including eicosapentaenoic acid (EPA; 20:5n − 3) and arachidonic acid (AA; 20:4n − 6), both being essential for normal larval development. The concentration of docosahexaenoic acid (DHA; 22:6n − 3) in oocytes was independent on muscle concentration, probably resulting from its selective transfer to the oocytes. The EPA/DHA ratio was highly conserved in sardine tissues, while DHA/AA and EPA/AA ratios varied significantly between samples. These results indicate that the FA content of eggs produced by sardines varies throughout the spawning season, egg FA concentrations decreasing as females lose condition, and FA composition also shows spatial variability. Both types of variability may have a significant impact on egg quality, particularly on the amount of reserves available to larvae affecting their resistance to starvation, and the appropriate FA composition required for normal growth.