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Antioxidant enzyme level response to prooxidant allelochemicals in larvae of the southern armyworm moth, Spodoptera eridania.

Larvae of the southern armyworm, Spodoptera eridania, are highly polyphagous feeders which frequently encounter and feed upon plants containing high levels of prooxidant allelochemicals. While ingestion of moderate quantities of prooxidants can be tolerated by these larvae, ingestion of larger quantities can result in toxicity. Studies were conducted to assess the role of the antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR) in the protection of S. eridania against redox active prooxidant plant allelochemicals. Dietary exposure of mid-fifth-instar larvae to either quercetin (a flavonoid) or xanthotoxin (a photoactive furanocoumarin), which generate superoxide radical, and singlet oxygen, respectively, resulted in an increase in SOD levels. CAT levels increased in all groups of S. eridania including control insects. This may have been due to the sudden exposure to food following an extended fast of 18 h (to insure that larvae would not reject the diet because of the prooxidants' bitter taste) with an eventual lowering of CAT values with time. GR activities did not significantly change except for a slight inhibition at the highest prooxidant concentrations used at 12-h post-ingestion. The data from these studies suggest that SOD responds to prooxidant challenges in these insects and together with CAT and GR contributes to the insect's defense against potentially toxic prooxidant compounds.

Animals↗

Toxicity of selected tremorgenic mycotoxins and related compounds to Spodoptera frugiperda and Heliothis zea.

A series of tremorgenic mycotoxins and related compounds were tested for oral toxicity to the fall armyworm (Spodoptera frugiperda) and corn earworm (Heliothis zea) by incorporation of materials into artificial diets and examining mortality and weights after 7 days. Significant mortality to both insect species was caused with dihydroxyaflavinine and roseotoxin B, while significant mortality to H. zea was also caused by penitrem A at 25 ppm. After 7 days, weighs of larvae treated with 25 ppm penitrem A, roseotoxin B, and verruculogen were less than 50% of controls for both insect species. Weights of H. zea larvae treated with 25 ppb of penitrem A were less than 50% those of control larvae. Relative toxicities of the tremorgens and related compounds to insects compared to vertebrates are discussed.

Animals↗

Ventral exocrine gland of larva of Spodoptera (Prodenia) litura (Fabricius) (Lepidoptera: Noctuidae).

The morphological and histological structure of the ventral exocrine gland of the larva of the tobacco caterpillar, Spodoptera (Prodenia) litura (Fab.) is shown. The proximal region of the gland is capable of protrusion through the prothoracic sternal slit as an elongate organ reaching upto the spinneret. The middle region of the gland shows characteristic giant cells in only one of the walls of the epithelium, while the distal region shows a thin flat epithelium with small round closely situated nuclei resting against the tunica propria. The gland probably secretes a gummy fluid for larval movements rather than a stinking odourous fluid for defense.

Animals↗

Sequence Analysis of the Bam HI-J Fragment of the Spodoptera litura Multicapsid Nucleopolyhedrovirus.

The BamHI-J fragment located at 25.8--9.9 map units of the Spodoptera litura multicapsid nucleopolyhedrovirus (SpltMNPV) genome was sequenced. The fragment contained four ORFs, one partial ORF potentially encoding C-terminal of chitinase gene and one partial homologous region (hr). The four ORFs included lef-8 gene, J domain protein gene (bjdp gene), ORF570 and ORF165. The ORF570 revealed 31% identity to the helicase-2 of Lymantria dispar MNPV. The ORF165 was unique to the SpltMNPV. The bjdp gene, reported here for the first time in baculoviruses, was one of J domain family protein genes, and the predicated amino acid sequence possessed a characteristic of J domain protein of other DnaJ proteins at its N-terminus. The lef-8 showed high identities to the homologs of reported baculovirus genomes. As a component of virus-encoded RNA polymerase, the LEF-8 of SpltMNPV had the conserved motif GIKICGIHGQKG near the C-terminal end. Analysis of the LEF-8 phylogenic tree demonstrated SpltMNPV was very closely related to SpliMNPV.

Journal Article↗

Nucleotide Sequence of a 4 730 Base Pairs Region of the Spodoptera litura Nucleopolyhedrovirus Genome.

A 4 730 bp region of the Spodoptera litura Nucleopolyhedrovirus genome EcoRI-E fragment was sequenced, in which the odv-e66 gene and other two ORFs, ORF 1086 and the p34 gene, were found. The SpltNPV odv-e66 gene open reading frame was deduced to be 2 079 bp, encoding a protein of 692 amino acids. Unlike odv-e66 genes of AcMNPV and LsNPV, only one transcriptional initiation conserved sequence ATAAG of the late gene was present upstream of the initial codon ATG of SpltNPV odv-e66 gene, and no poly(A) signal was detected at the 3' end of the gene. Four highly conserved regions homologous to those of LdNPV, LsNPV, AcMNPV, BmNPV, CfNPV and OpNPV were present in the amino acid sequence of SpltNPV ODV-E66 protein, with the putative nuclear targeting signal(NTS) KKIWAEDGR in the fourth conserved region. The ORF upstream of the odv-e66 gene had the same direction of odv-e66 gene, but different reading frame. Besides the CAAT box, the early gene regulatory element ACGT and GC motifs and the transcriptional initiation conserved sequence TTAAG of the late gene were present at the 5' end of this ORF. A putative protein of 289 amino acid residues with Mr. about 34 kD was encoded by the ORF, so that it is named P34 protein. A leucine zipper and a leucine zipper-like structure were found in the C- and N-termini of SpltNPV P34 protein respectively. The functions of those regulatory elements may be of great importance to SpltNPV genome. An 1 086 nt open reading frame (ORF) capable of encoding a 361 amino acids polypeptide was found at the 3' end of the odv-e66 gene and in opposite direction. The ORF 1086 protein shares 44% amino acid sequence homology with AcMNPV ORF 109.

Journal Article↗

Cloning and Sequence Analysis of the Chitinase Gene of Spodoptera litura Nuclear Polyhedrosis Virus.

Using AcMNPV chiA-containing fragment as a probe, the chitinase gene of Spodoptera litura nuclear polyhedrosis virus (SpltNPV) was localized in two contiguous fragments, XbaI 5.1 kb and 2.1 kb, and the intact SpltNPV chiA gene was obtained by cloning and sequencing those two fragments. The complete open reading frame of the gene was 1 695 nucleotide long, encoding a putative protein of 564 amino acids with molecular weight of 62.9 kD. Sequence analysis further revealed that the 5' noncoding region had a baculovirus late promoter motif TAAG. A polyadenylation signal, AATAAA, was located dowmnstream of the translation stop codon. A putative signal peptide was present at the N-terminus of the protein. This gene shares 57% homology with AcMNPV chitinase gene.

Journal Article↗

The NADP-dependent methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate cyclohydrolase-formyltetrahydrofolate synthetase is not expressed in Spodoptera frugiperda cells.

The insect cell line derived from Spodoptera frugiperda (Sf9) does not express the activities of the trifunctional NADP-dependent methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate cyclohydrolase-formyltetrahydrofolate synthetase. The lack of synthetase activity was confirmed by the inability to incorporate radiolabeled formate into nucleotides. The cells express, instead, a Mg2+ and NAD-dependent bifunctional methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate cyclohydrolase with properties similar to the enzyme found in the mitochondria of transformed mammalian cells. In contrast, the enzyme in Sf9 cells is localized in the cytoplasm. Nutritional studies in defined medium with dialyzed serum demonstrated that the Sf9 cell does not required added purines or pyrimidines for growth. It is auxotrophic for cysteine and glycine; this latter requirement is probably due to the absence of mitochondrial serine hydroxymethyltransferase. Incorporation of labeled glycine and serine into DNA indicates that only serine is a source of one-carbon units. These results suggest that the mitochondria in Sf9 cells do not play a major role in folate-mediated metabolism.

Aminohydrolases↗

[Realized resistance heritability and resistance risk of spodoptera exigua to cyhalothrin, fenvalerate and alpha-cypermethrin].

In this paper, Spodoptera exigua from Fuzhou of Fujian Province was continuously selected and bred to test its resistance to cyhalothrin, fenvalerate and alpha-cypermethrin, and to evaluate its realized resistance heritability (h2) and resistance risk. The results showed that after 12, 10, and 10 generations, the resistance of S. exigua to cyhalothrin, fenvalerate and alpha-cypermethrin was increased by 18.1-, 27.6-, and 45.4-fold, and the realized cyhalothrin, fenvalerate and alpha-cypermethrin was increased by 18.1-, 27.6-, and 45.4-fold, and the realized resistance heritability was 0. 2567, 0. 3571 and 0. 4239, respectively. Assuming that the h2 of field S. exigua population was half of these values, it required 9 to approximately 20 generations for cyhalothrin, 6 to approximately 14 generations for fenvalerate, and 5 to approximately12 generations for alpha-cypermethrin to obtain 10-fold increase in resistance under selective pressure of 50% to approximately 90 % mortality for each selective generation. Of the three pyrethroids, the resistance risk to cyhalothrin was smaller than that to fenvalerate and alpha-cypermethrin.

Animals↗

Comparative studies on the development of the entomogenous nematodes Steinernema feltiae Filipjev on Spodoptera littoralis (Boisduval) and Musca domestica Linneous.

Comparative studies were made at 26 degrees C under laboratory conditions on the development of Steinernema feltiae Filipjev (= Neoaplectana carpocapsae) on the different stages of Spodoptera littoralis (Biosduval) and Musca domestica (Linneous). In the present investigation the host species influence manifested itself, in a quicker development rate along with the cotton leafworm S. littoralis. Particular development stages took place earlier in the S. littoralis than in M. domestica. As far as the occurrence dates of particular stages of S. feltiae were concerned, the differences in relation to the two host species were significant. The changes in numbers of giant forms in the development course of populations in the caterpillars, pupae and imagos body of both species were studied. The numbers of giant forms correspond with the numbers of invasive larvae, it seems that at first they are growing and then decreasing.

Animals↗

Expression of biologically active human antithrombin III by recombinant baculovirus in Spodoptera frugiperda cells.

Antithrombin III (ATIII) is a plasma-borne serine protease inhibitor that plays a pivotal role in the regulation of hemostasis. The cDNA for ATIII has been available, but genetic studies on the functional domains of ATIII have not progressed because of the absence of an expression system that will yield sufficient quantities of biologically active protein for biochemical analyses. In the present studies the cDNA of the human antithrombin III gene was inserted into the vector pVL 1393, which is suitable for cotransfection of Spodoptera frugiperda (Sf9) insect cells with Baculovirus wild-type DNA. Recombinant virus particles were selected by the presence of occlusion-negative plaques. Upon infection with purified recombinant virus, Sf9 cells secreted 10-35 micrograms of ATIII/1 x 10(6) cells. Southern analysis of DNA from infected cells demonstrated incorporation of the full-length cDNA into the Baculovirus recombinant, and RNase protection experiments verified the presence of full-length transcript. This recombinant ATIII protein was immunologically reactive with antisera raised against native human ATIII, formed stable complexes with thrombin, and was heparin-accelerated at the same concentration as native human ATIII. In addition, the recombinant ATIII retained specificity for the same molecular species of heparin that activates authentic human ATIII. This is the first successful production of active, recombinant ATIII in quantities that will allow purification on the milligram scale and permit a biochemical analysis of genetically engineered variants.

Amino Acid Sequence↗

Sex pheromone biosynthetic pathway in Spodoptera littoralis and its activation by a neurohormone.

Deuterium-labeled fatty acids have been used to elucidate the sex pheromone biosynthetic pathway in Spodoptera littoralis. Label from palmitic acid was incorporated during the scotophase into all the pheromone acetates and their corresponding fatty acyl intermediates. (Z,E)-9,11-tetradecadienyl acetate, the major component of the pheromone blend, is synthesized from palmitic acid via tetradecanoic acid, which, by the action of a specific (E)-11 desaturase and subsequently a (Z)-9 desaturase, is converted into (Z,E)-9,11-tetradecadienoate. By further reduction and acetylation, this compound leads to the dienne acetate. Deuterated precursors applied to the pheromone gland during the photophase were also incorporated into the pheromone. The percentage of labeled (Z,E)-9,11-tetradecadienyl acetate relative to natural compound was significantly higher during the light period. Label incorporation from different intermediates into the pheromone was stimulated by injection of brain-subesophageal ganglion extract during the photophase. The influence of the pheromone biosynthesis-activating neuropeptide on the biosynthetic pathway is discussed.

Animals↗

Different pathways of DNA synthesis are operative in the fat body of Spodoptera litura (insecta) during larval and pupal development.

In Spodoptera litura, the DNA content of the fat body increases during the last larval instar. Radiolabelling studies reveal high degree of H3-thymidine incorporation in the fat cell DNA, during last larval stage of development. At prepupal stage the DNA synthesised during larval development is degraded, at least partially and the resultant nucleotides are most likely used as building blocks for the fat cell DNA synthesis during pupal development and pupal-adult metamorphosis, because very little H3 -thymidine injected into pupae is incorporated in DNA, though in fact, significant amount of DNA is synthesised during this period.

Animals↗

The influence of infection with Steinernema feltiae Filipjev on the haemocyte picture of the cotton leaf worm (Spodoptera littoralis Boisduval).

Infection of Spodoptera littoralis Boisduval larvae with Steinernema feltiae Filipjev nematodes caused sharp variation in proportional percentages of differential haemocyte counts. The mean total haemocyte count per mm3 of haemolymph in control larae of S. littoralis was 6000 haemocyte mm3. After infection with 50, 100, 200 and 500 parasitic nematodes of S. feltiae, the mean total count was significantly increased to 17,250, 18,600, 21,000 and 24,000 haemocyte mm3 respectively.

Animals↗

A high density field population of mixed phases (solitaria and gregaria) of armyworm (Spodoptera exempta) caterpillars in Kenya.

An unusual type of mass propagation ("outbreak") of armyworms (Spodoptera exempta) was found during the rainy season in the highlands of central Kenya. In a very small area it had a high density of individuals but - unlike normal outbreaks - it comprised a continuity of different developmental stages from young caterpillars to moths. The older caterpillars were of phase solitaria or gregaria together with numerous individuals of intermediate forms. From their different ages it is improbable that they all came from parental moths having arrived together. The finding of such type of outbreak seems of interest for understanding the development of the huge uniform outbreaks of this species frequently occurring in East Africa.

Animals↗

Role of the perineurium and glial lacunar system in nutrition and storage of nutrients in the central nervous system of Spodoptera litura Fabr. (Lepidoptera: Noctulidae) at the time of reorganisation of the neural lamella during metamorphosis.

Brains and nerve cords of Spodoptera litura (Fabr.) of various stages during metamorphosis were tested for acid mycopolisaccharides (AMPS), PAS-positive substances, glycogen, proteins and lipids. During reorganisation of the neural lamella in pupal period glycogen and PAS-positive substances are stored in perineurium and AMPS in the glial lacunar system (GIS) and below the perineurium as a thin layer. Lipids and AMPS diffuse in the GIS. The perineurium and GIS serve for the passage as well as storage of nutrients. In neuropile, only proteins are present. Significances of these findings is discussed.

Animals↗

Intoxication of cattle on kikuyu grass following army worm (Spodoptera exempta) invasion.

Clinical features and pathological and mycological findings in a field outbreak of intoxication in dairy cattle grazing kikuyu grass are reported. The outbreak followed invasion of the grass by the army worm (Spodoptera exempta). Clinical signs included drooling of saliva, depression, apparent inco-ordination, sunken eyes, ruminal distension and atony, recumbency, moderate diarrhoea and "sham drinking". Seventy-seven cows (64%) were clinically affected over a period of 12 days. Of these, 37 died. Necropsies performed on 4 affected cattle revealed necrosis of the epithelium of the forestomach, which was consistently more severe in the omasum. Light microscopy showed extensive necrosis of the epithelium of the forestomach with associated fibrinopurulent inflammation. The stratum spinosum and s. granulosum were selectively involved, but the s. basale was generally preserved. Electron microscopical examination of ruminal and omasal epithelium from 2 of these cattle revealed cytopathological features in the s. spinosum and s. granulosum which were consistent with stages in an acute, anoxic type of injury. Mycological examination of the pastures revealed sparse growth of a mixed fungal population, which included Myrothecium verrucaria. There was no evidence of heavy fungal infestation. Previous evidence that M. verrucaria, or other fungi, may be involved in the aetiology of kikuyu grass poisoning of cattle in New Zealand is addressed. It appears improbable that any of the fungi isolated in this investigation could have played an important role in the aetiology of this outbreak.

Animals↗

Baculovirus infection does not alter N-glycosylation in Spodoptera frugiperda cells.

Spodoptera frugiperda cells are the standard host system for baculovirus-vector-mediated expression of recombinant glycoproteins. In an attempt to explore their ability to produce complex N-glycans containing terminal neuraminic acid, we tested both mock- and AcMNPV-infected SF9 cells. To elucidate the structures of the carbohydrate chains of cellular glycoproteins, radiolabeled oligosaccharides were liberated by treatment with endo-H and glycopeptidase F. When the endo-H resistant material was subjected to sequential degradation with exoglycosidases, only truncated carbohydrates with the structures Man3GlcNAc2 and Man3[Fuc]GlcNAc2 were found. There was no evidence for the presence of neuraminic acid and complex N-glycans. The results indicate that SF9 cells have only a limited capacity to process N-glycans. Infection with AcMNPV has no significant effect on glycosylation in these cells.

Amidohydrolases↗

Postembryonal development of the testes in cotton leaf worm, Spodoptera littoralis (Boisd.) (Noctuidae, Lepidoptera).

During postembryonal development of males of Spodoptera littoralis the paired four-follicular larval testes undergo fusion and torsion, forming in the prepupal stage one gonad composed of eight testicular follicles. From the 6th larval till early pupal stage, the interior of the testicular follicles is divided into the following zones: 1) germarium with apical complex (an apical cell and two kinds of spermatogonia); 2) a zone, in which the single spermatogonia become surrounded by somatic cells, thus forming spermatogonial cysts; 3) a zone in which the spermatogonia inside the cysts undergo six incomplete mitotic divisions to form a syncytium of 64 spermatocytes (eupyrene spermatocytes with spherical nuclei or apyrene ones with polymorphic nuclei); 4) a zone, in which the spermatocytes transform into eupyrene or apyrene spermatids (256 per one cyst). In the mid-period of pupal stage two events occur: the apical cell in germarium degenerates and the eupyrene spermatogenesis ends. The apyrene spermatogenesis starts in the 6th larval instar and ends in the late pupa. In the late pupal and young imago testis, apyrene spermatozoa cysts form a compact layer under the gonadal wall, whereas the eupyrene cysts are loosely scattered in the central region of testicular follicles. The flagellum of the eupyrene spermatozoon is characterized by one mitochondrial derivative, by axonemal microtubules containing electron-dense material and by two kinds of appendage structures on the surface: lacinate appendages and satellite bodies. The flagellum of apyrene spermatozoon possesses two mitochondrial derivatives. It has neither extracellular appendages nor electron-dense material in microtubules. In seminal follicles the apyrene spermatozoa acquire a thick coating exhibiting periodic structure.

Animals↗