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Apoptosis of Spodoptera litura cells induced by AcMNPV ie-1 gene.

Apoptosis was induced in Spodoptera litura Sl-zsu-1 cells transfected with plasmid pAcie-1 DNA containing ie-1 gene of Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV). Apoptotic bodies appeared 24 h post transfection and cell mortality peaked after 48 h. Electrophoresis of total DNA extract of Sl-zsu-1 cells showed a DNA ladder, indicating that transfection of ie-1 gene alone can trigger apoptosis directly or indirectly. It was shown by cytochalasin D (CD) and ammonium chloride inhibition assays that the entry of virions into cytoplasm was required for induction of apoptosis. Reverse transcriptase PCR (RT-PCR) was used to demonstrate that ie-1 gene had transcribed in Sl-zsu-1 cells immediately after infection. Inhibition of DNA replication could not block the cell death process, suggesting that this apoptosis was independent on viral late replication events. Apoptosis did not occur when Sl-zsu-1 cells were infected with an AcMNPV mutant tsB821 at its nonpermissive temperature. Therefore, we concluded that the ie-1 gene is at least one of the factors inducing apoptosis in Sl-zsu-1 cells infected with AcMNPV.

Animals↗

Insecticidal activity of selected actinomycete strains against the Egyptian cotton leaf worm Spodoptera littoralis (Lepidoptera: Noctuidae).

Actinomycete strains isolated from soil samples of Saudi Arabia and Egypt were used in the present study to investigate the biological activity of their secondary metabolites on the cotton leaf worm Spodoptera littoralis. Secondary metabolites of 41 strains were given to the newly molted last instar larvae through the food plant (Castor leaves). Many actinomycete strains (58% of all) caused larval mortality ranging from 10-60%. Treatments with strains no. 7, 36, 41 resulted in significant prolongation of larval life (10.0 +/- 1.26, 11.5 +/- 0.7 and 10.6 +/- 1.5 days Vs 9.2 +/- 0.4 of controls). Pupation was affected by the actinomycete treatments (60.1% of all). Success of pupation ranged only from 20 to 80%, depending on the efficiency of the tested secondary metabolites. In contrast, the pupal duration had not been significantly affected but 19.5% of actinomycete strains halted the pupation, because different percentages of pupal deformities were counted (100% pupal deformation after treatment with secondary metabolites of actinomycete strains no. 36). In addition, considerable lethal effect of some actinomycetes were observed on pupae (60, 42.9, 40, 50 and 66.6% pupal mortalities caused by secondary metabolites of strains: 2, 9, 16, 18 and 26, respectively). Secondary metabolites of 34.1% of all actinomycete strains blocked the adult emergence. Depending on the available data, Streptomyces and Streptoverticillum were found the most potent actinomycetes affecting the biological and physiological criteria of the present insect species.

Actinomycetales↗

Enzymatic changes and cytotoxic effects of Metarhizium anisopliae fungal extracts on the Egyptian cotton leaf worm Spodoptera littoralis (SL 96) cell line.

The extra-and intra-cellular extracts of two Metarhizium anisopliae isolates: Ma23 and MR3, were introduced to a cell line of Spodoptera littoralis. The cell line (Sl96) was established in the Entomovirology centre from S. littoralis ovaries. MR3 extracellular fluid-treated culture had an increase in the Acid phosphatase (AcP) level as compared either to the untreated cell line or to the fungal extract alone. Moreover, the enzymatic tests showed that alpha and beta-glucosidases have been detected only after the treatment with extracellular fluid of MR3. The elevation of AcP level that appeared after the treatment of MR3 coincided with a conspicuous decrease of about one third of the initial number (96 x 10(3)) of the Sl96 cell. vacuolization and lysis of some cells. Deformation of the cells and shrinkage of cell membrane have been observed in the case of Ma23 treatments. Crystal formation was restricted to the treatment of MR3 extracellular extract. The results are discussed in the light of the possibility that the cytotoxic effects and enzymatic changes are attributed to the virulence of the fungal isolates.

Animals↗

[Preliminary studies on the apoptosis of Spodoptera litura cells induced by Two baculoviruses].

Microscopic examination of Spodoptera litura(SL-1) cell infected with AcMNPV or SfaMNPV revealed progressive cell blebbing starting at 8 h-12 h postinfection and culminating in total cell destruction at 24 h postinfection. The fragmentation of the infected cell nuclei was observed by stained with the specific fluorescent dye DAPI. Agarose gel electrophoresis analysis of the DNA extracted from infected cells show typical DNA ladder. All these supported that both viruses infected SL-1 cells undergo apoptosis. Through lipofectin transinfection, we observed that only the DNA of AcMNPV or SfaMNPV can also induce apopotosis of SL-1 cell. Virus titer analysis revealed that neither viruses can duplicate in SL-1 cells.

Animals↗

Use of RAPD in assessing the genetic variability in Spodoptera litura.

A polymerase chain reaction based assay to distinguish six different ecotypes of the Spodoptera litura, a sporadic insect pest and a predominant defoliator of various crops was carried out. A total of 40 random primers were screened to reveal the existence of polymorphism between the populations. Among them eight showed scorable banding patterns and three primers (OPA-01, OPA-05, OPM-01) exhibited distinguishable banding patterns. However, Chengalpattu and Chennai populations revealed their closed relatedness and Coimbatore population stood distantly from others.

Animals↗

Evidence for negative cross resistance to insecticides in field collected Spodoptera littoralis (Boisd.) from Lebanon in laboratory bioassays.

A sample of Spodoptera littoralis pupae were field collected from in Lebanon in November 1999. Approximately 50 healthy pupae were used to initiate a laboratory colony (stain code LEB). As a benchmark for sensitivity to insecticides the susceptible laboratory strain (LET) was used for comparison. The LET stain has been held in laboratory culture for over 10 years and originated from Israel. Three test chemicals were investigated; Spinosad (Tracer, 480 g/L SC), chlorpyrifos (Dursban 480 g/L EC) and cypermethrin (Ambush C 100 g/L EC) representing a range of different modes of actions. To estimate the sensitivity of the LEB strain relative to the LET stain laboratory bioassays, conducted on second instar larvae exposed treated tomato leaves for two days. The method used was based on IRAC (Insecticide Resistance Action Committee) insecticide susceptibly test number 7. Larvae from the LEB strain were tested in both the F1 and F2 laboratory generations. LC50 values were calculated using a Log-Probit model allowing the susceptibly of each strain to each test item to be compared. The LEB (field) strain were approximately twice as sensitive to spinosad compared with the LET (laboratory) strain. The LEB strain was 6-10 times less sensitive to chlorpyrifos which was within the range expected. However the LEB stain was 250-360 times less sensitive to cypermethrin compared with the LET strain, indicating a high level of resistance to pyrethroids in the field population. This kind of relationship gave a strong indication for negative cross resistance between spinosad and cypermethrin. It was concluded that spinosad had potential as a resistance management tool due to its novel mode of action and negative cross resistance with pyrethroids.

Animals↗

[Influence of host age on the epizootic of nuclear polyhedrosis virus of Spodoptera litura].

Through investigation of tracking infected Spodoptera litura larvae, a study was carried out to observe the epizootic dynamics of nuclear virus disease of S. litura after virus release in different instar with concentration of 2.15 x 10(7) PIBs.ml-1 in vegetable field. The results showed that with the age of S. litura increasing, the susceptibility to virus, mortality and death velocity decreased, the disease death sustained period of larvae obviously elongated, and the time that larval mortality and diseased prevalence began and peaked were postponed. The distribution of disease death time could be well simulated by Complementary log-log model (CLL). Hosmer-Lemoshow test showed that the theoretic values well fitted to observed data, and t-test indicated that the coefficients of the model reached significant level. The distribution of diseased prevalence of host larvae was simulated by Holliday model, the regression of function was significant by F-test, and its coefficients reached or approached significant level. Moreover, the cumulative diseased death time distribution could be described by S-type models (Gompertz, Logistic, Weibull), in which, the Gompertz model was the best.

Age Factors↗

[Cooperative effect of Spodoptera litura nuclear polyhedrosis virus with parasite wasp, Microplitis sp. on the S. litura larvae].

The relationship between the survival rate, developmental duration of Microplitis sp. in Spodoptera litura larvae and time of exposure of the host larvae to virus, the inoculation virus concentration were investigated to determined if parasites which emerged from virus-infected S. litura larvae had normal survive, and if the parasites could sever as the vector for S. litura nuclear polyhedrosis virus (SINPV). There was no significant difference in time that parasites spent in virus-infected larvae and healthy one, indicating the SINPV did not affect the developmental period of Microplitis sp. Parasite larvae in hosts exposed to virus after parasitization compmeted their development before their hosts died of virus infection. The proportion of parasites survived varied with the inoculation virus concentration and the time of exposure of the host larvae to virus, the percent of parasites survived increased as the time between host parasitization and host virus exposure increased, and decreased as the inoculation virus concentrations increased. The female parasites which developed or oviposited in virus-infected hosts, and manually contaminated the ovipositor of the females with virus suspension could all carry infective doses of the virus, and transmit virus from infected larvae to healthy one through ovipositor. A female parasite which developed or oviposited in virus-infected hosts transmitted infective doses of the virus to an average of 2.14, 2.45 healthy host larvae exposed to it respectively. A female parasite from the cocoon contaminated with SINPV of different virus concentration could transfer infective doses of the virus to an average of 1.45 healthy host larvae. A female parasite exposed to the mixture different virus concentration and 10% honey-water solution could also transmit virus to an average of 0.94 larva.

Animals↗

Performance of Spodoptera litura Fabricius on different host plants: influence of nitrogen and total phenolics of plants and mid-gut esterase activity of the insect.

Five host plants [castor, Ricinus communis (Carolus Linnaeus); cotton, Gossypium hirsutm (Carolus Linnaeus); tomato, Lycopersicum esculentum (Philip Miller); mint, Mentha arvensis (Carolus Linnaeus) and cabbage, Brassica oleracea (Carolus Linnaeus)] belonging to different families were used to study the performance of the Asian armyworm, Spodoptera litura larvae. Highest consumption of food and dry weight gain was observed in larvae fed on castor. Mint did not support optimum larval growth because of low digestibility and low efficiency of conversion of digested food to body matter. Dry weight gain ranged from 26.64 mg on mint to 86.80 mg in castor. These differences tend to be related to nitrogen and total phenolics content of the leaf tissues; however, the most clear-cut correlation is an inverse one between the host plant preference and the ratio of total phenolics to nitrogen in the leaf tissues. Mid-gut esterase activity in larvae showed an increasing trend with the increase in total phenolics: nitrogen ratio in the test plants and the order of mid-gut esterase activity in larvae was mint > cabbage > cotton > tomato > castor.

Animals↗

The hemolymph of caterpillars Spodoptera littoralis: physico-chemical properties and ionic composition compared to culture media.

In this paper, we determined some physico-chemical properties like osmotic pressure, pH and electrical conductivity of the hemolymph from caterpillars of Spodoptera littoralis (Lepidoptera: Noctuidae) during the last larval instar. It was of interest that we observed an increase in osmotic pressure with the increase in age in the last instar that may concur with the start of histolysis at metamorphosis. These physicochemical properties were then compared to those of Grace's and modified Grace's tissue culture medium. In addition, concentrations of the cations Na, K, Ca and Mg, and the anions Cl, NO3, PO4 and SO4 were determined in the insect hemolymph of S. littoralis. The cations K and Mg reached high values with a percent of about 52% of the total amount of cations. The concentration of sodium was low. The total sum of the anions consisted about 56 meq/1, and this allows to neutralise about 45 % of the total cations.

Animals↗

Alzheimer amyloid precursor aspartyl proteinase activity in CHAPSO homogenates of Spodoptera frugiperda cells.

Presenilins are polytopic, integral proteins that control intramembranous proteolysis at the "gamma-" and "epsilon-" cleavage sites of the Alzheimer amyloid-beta precursor protein (APP) to yield amyloid-beta peptide (Abeta) and the APP intracellular domain (AICD). We have overexpressed a constitutively active, pathogenic form of PS1 (known as PS1 Delta exon 9) together with its substrate, APP-C99, in Spodoptera frugiperda (Sf9) cells. Sf9 cells have been reported to lack endogenous gamma-secretase, an unexpected finding since there exists an insect homologue of PS1. In our hands, neither intact insect cells coexpressing PS1 Delta exon 9/APP-C99 nor the aqueous homogenates of these cells displayed obvious products of the gamma- or epsilon-secretase reactions, as reported. Surprisingly, when APP-C99-expressing cells were homogenized in 3[(3-cholamidopropyl) dimethylammonio]-2-hydroxypropanesulfonic acid (CHAPSO), a detergent known to support gamma-secretase activity, subsequent incubation led to the accumulation of an AICD-like peptide (AICD-L). Aspartyl proteinase inhibitors were effective in preventing the appearance of AICD-L, but inhibitors of other classes of proteinases were ineffective. Immunoprecipitation-mass spectrometry of AICD-L revealed its identity as the minor of the two known AICDs.

Alzheimer Disease↗

[Establishment of Spodoptera exigua multicapsid nucleopolyhedrovirus BAC-TO-BAC expression system].

Present studies describe the successful establishment of Spodoptera exigua multicapsid nucleopolyhedrovirus (SeMNPV) BAC-TO-BAC expression system. The mini-F-lacZ-attTn7-kan fragment (Luckow et al, 1993) was inserted into SeMNPV US1 isolate (SeUS1) at polyhedrin gene locus by directly cloning. The recombinant virus containing low-copy-number mini-F replication, which named bacmid, can propagate in Escherichia coli. Because SeUS1 isolate is make up of several genotypes and one bacmid carries one SeMNPV genotype, the SeUS1 BAC library is established by all SeMNPV bacmids (SeBAC). REN analysis for 111 SeBAC shows that SeUS1 consists of the genotype with whole SeMNPV genetic information and several genotypes with various different deletions. Progeny virus can be produced in insect cell line after transfection with SeBAC10, which carries the whole SeMNPV genome. So SeBAC10 is a shuttle vector that can replicate in eukaryocyte as well as prokaryocyte. Considering the insert mutation of SeMNPV polyhedrin gene (Seph) in SeBAC10, Seph was reintroduced into the bacmid by site-specific transposon-mediated insertion at attTn7, the target site for the bacterial transposon Tn7. The derived recombinant SeBAC10 was named SeBAC10ph. After SeBAC10ph was transfected into Se301 cells (a susceptible insect cell line to SeMNPV), cytopathogenic effect was shown and polyhedra appeared, which indicate that the foreign gene (Seph) is expressed.

Animals↗

Establishment and characterization of Spodoptera littoralis cell line (SL 96), adapted at 19 degrees C permissive for different baculoviruses.

Cells from ovarioule tissues of the Egyptian cotton leaf worm, Spodoptera littoralis (SL 96) were established, identified and characterized comparing with other lepidopteran cell lines using isozymes, karyotyping, RAPD PCR, and polyacrylamide gel electrophoresis polypeptide PAGE profile. Comparison of the number of chromosomes and PAGE profiles; also the isozymes profiles demonstrate the possibility to distinguish the cell lines by those analyses and having a distinguishable finger print for every cell line. A complete replication of the S. littoralis granulovirus (SpliGV), nucleopolyherovirus (SpliNPV), and Phthorimaea operculella granulovirus (PhopGV) was obtained in vitro by both viral infection and DNA transfection in the S. littoralis (S196) cell line. Spli-GV and PhopGV multiplied extensively during several passages in S196 cells at 19 degrees C. Virus infected cells were incubated at 19 degrees C for the first 4 hours and then at .27 degrees C for the rest of the experiment (20 days). Generally, all the three viral multiplications proceeded at the same rate approximately. Comparison of Spli GV progenies multiplied either in vivo or in vitro, using electron microscopy and restriction profile analysis, showed their identity. Viral infections were detected also by specific prepared nucleic probes.

Animals↗

[Antifeeding effects of crude lantadene from Lantana camara on Plutella xylostella and Spodoptera litura larvae].

In this study, crude lantadene was extracted from Lantana canmara leaves, and its antifeeding effects on Plutella xylostella and Spodoptera litura larvae were tested. In no-choice test, crude lantadene at 1.6 mg x ml(-1) concentration had antifeeding effects on the 2nd instar P. xylostella larvae and 1st instar S. litura larvae, with the antifeeding rate being 62.4% and 33.1%, respectively within 48 h. In choice test, even a low concentration (0.4 mg x ml(-1)) crude lantadene still had anti-feeding effects on the 2nd instar P. xylostella larvae, and the antifeeding rate at 0.4, 0.8 and 1.6 mg x ml(-1) concentration was 52.7%, 55.5% and 78.9%, respectively. Crude lantadene only at 1.6 mg x ml(-1) concentration had anti-feeding effects on the 1st instar S. litura larvae, and the antifeeding rate was 33.0%. For the 2nd instar S. litura larvae, crude lantadene had no antifeeding effects both in no-choice and in choice test.

Animals↗

Evaluation of substituted oxime ethers for growth regulatory activity against Spodoptera litura (F.).

Insect growth regulatory activity (IGR) of fifty-two substituted oxime ethers were evaluated against an important polyphagous lepidopteran crop pest, Spodoptera litura (F.). A number of compounds produced symptoms comparable to exogenously applied juvenile hormone. Maximum IGR activity was exhibited by 4'-(2,6,6-trimethyl-2-cyclohexen- -yl)-3'-buten-2'(E)-ketoxime-N-O-alkyl ether with an ED50 (morphological) of 40 microg g(-1) body weight, compared to 20 microg g(-1) of JH III. Two more compounds namely 4'-(2,6,6-trimethyl-2-cyclohexen-1-yl)-3'-buten-2'(Z)-ketoxime-N-O-methyl propyl ether (ED50 192 microg g(-1)) and 4'-(2,6,6-trimethyl-1-cyclohexen-1-yl)-3'-buten-2'(E)-ketoxime-N-O-pentyl ether (ED50 380 microg g(-1)) showed considerable IGR activity, whereas 4'-(2,6,6-trimethyl-2-cyclohexen-1-yl)-3'-buten-2'(E)-ketoxime-N-O-pentyl ether was found to be toxic to the larvae (ED50 268 microg g(-1)). Three compounds used in this study were also synergised by piperonyl butoxide (PBO). The synergistic ratios were found in the range of 1.33 to 4.605. The ovicidal activity of the oxime ethers is not significant.

Animals↗

Effect of JH III and substituted oxime ethers on the in vitro protein and RNA synthesis in male accessory reproductive gland (MARG) of Spodoptera litura (F.).

The physiological action of two substituted oxime ethers namely: 4'-(2,6,6-trimethyl-2-cyclohexen-1-yl)-3'-buten-2'(E)-ketoxime-N-O-propylether (compound No. 3) and 4'-(2,6,6-trimethyl-1-cyclohexen-1-yl)-3'-buten-2'(E)-ketoxime-N-O-pentylether (compound No. 34,) were compared with that of JH III in an in vitro assay to monitor the synthesis of RNA and protein in male accessory reproductive gland (MARG) of Spodoptera litura by using 3H-leucine and 3H-uridine, respectively. Both the compounds have stimulated protein synthesis compare to control. Compound No 34 is slightly more effective than JH III in increasing the protein synthesis at physiological concentration of 10(-6) and 10(-5) M. Compound No 3 and JH III have doubled the RNA synthesis and increased the protein synthesis by 1.5 times over the control at 10(-4) to 10(-6) M concentrations. While JH III at 10(-5) M significantly enhanced RNA synthesis, similar effect is produced only at 10(-3) M by compound No 3 and 34.

Animals↗

Protein profiles of Spodoptera litura (F.) male accessory reproductive gland and its secretions.

Protein profiles of the male accessory reproductive glands of a polyphagous pest, Spodoptera litura (Lepidoptera: Noctuidae) and their glandular secretions were analyzed by electrophoresis under denaturing conditions. The study revealed the presence of 23 proteins in the glands and 14 in the secretions. The molecular weights of the gland proteins ranged from 163 to 3.8 KD whereas that of secretions ranged from 100 to 3.8 KD. The 9 low molecular weight proteins of the secretions are a characteristic feature. It is suggested that the secretions may contain a sex peptide that has a role in the fertility and fecundity of the females.

Age Factors↗

[Modeling of Spodoptera litura nuclear polyhedral virus epidemic at different temperatures].

The study on the nuclear polyhedral virus epidemic of Spodoptera litura under effects of temperature showed that at above 29 degrees C, S. litura larvae started to die 2-3 d after virus-inoculation, and reached the peak 4-5 d after the inoculation. The mortality and death velocity of virus-infected host larvae were increased with increasing incubation temperature. No thermo-inhibition temperature for the virus was observed in the test temperature range, while there existed a constant temperature range, i.e., 29-35 degrees C, for the highest mortality of infected host larvae. Complementary log-log regression model could well simulate the daily distribution of host larvae mortality, and the cumulative disease death time could be described by Gompertz model. The revised biophysical model could well simulate the relationship between S. litura larvae mortality and temperature, and be applied to virus epidemic forecasting.

Animals↗