Induction of chlorosis, ROs generation and cell death by a toxin isolated from Pyricularia oryzae

The ethyl acetate extract of the conidia germination fluid from an Avena isolate (Br58) of Pyricularia oryzae had chlorosis-inducing activity on oat leaf segments. The same activity was also present in the acetone extract of an oatmeal agar culture of Br58. Fungal cultures were used for a large-scale preparation. A series of acetone and ethyl acetate extraction monitored by chromatography was used to isolate an active fraction. The active principle was purified by HPLC. We show by NMR and LC/MS that the toxin was an oxidized C18 unsaturated fatty acid named Mag-toxin. Mag-toxin induced chlorosis on oat leaf segments incubated in the light but not in the dark. Reactive oxygen species (ROS) and cell death were induced by Mag-toxin in oat cells. The sub-cellular localization of ROS generation induced by the toxin treatment was correlated with the location of mitochondria. Interestingly, the induction of ROS generation and cell death by Mag-toxin was light-independent.     

Antimicrobial activity and phytochemical screening of Ocimum americanum L extracts against pathogenic microorganisms

The aim of the present study is to assess the antimicrobial activities of various leaf extracts of Ocimum americanum were tested against pathogenic microorganisms. Preparation of different extracts viz., aqueous, acetone, ethyl acetate and methanol through soxhlet extraction method. Various extracts were investigated against MTCC strains of Bacillus cereus, Clostridium penfrigens, Klebsilla pnemoniae, Salmonella paratyphi, Candida albicans and Aspergillus niger by agar well diffusion and disc diffusion methods. Minimum inhibitory concentration (MIC), Minimum Bactericidal/Fungicindal Concentration (MBC/MFC) were determined through micro dilution method. Elucidation of phytochemicals and functional groups were observed by HPLC and FT-IR respectively. Ethyl acetate leaf extract of O.americanum showed significant antimicrobial activity against the all tested pathogens in agar well diffusion method in which B.cereus (17 mm) was observed high zone of inhibition. Whereas lowest inhibition was observed in aqueous extract against C.pentrigens (7 mm). The ranges of MIC values from 0.78 μg/ml to 50 μg/ml and MBC/MFC 1.56 μg/ml to 50 μg/ml were observed. Phytochemicals such as alkaloids, steroids, saponins, flavonoids, tannins, terepenes, phenolic compounds cardiac glycosides were detected. Saponinns, flavonoids, tannins, phenolic compounds were observed in only ethyl acetate leaf extracts. Functional group of the leaf extracts was exhibited by FTIR and HPLC analysis of the ethyl acetate leaf extract was elutated at six peaks. Based on the results we concluded that ethyl acetate leaf extract of O.americanum has proved to be potentially effective than the other extracts. Therefore, ethyl acetate leaf extract of O.americanum could act as antimicrobial agent and further studies are recommended for isolation of compounds and toxicological studies.     

树脂吸附法分离瓜果腐霉毒素中的除草活性物质

本试验以对禾本科杂草具有除草活性的瓜果腐霉菌株PAM1为试验材料,研究了X-5、AB-8,H-103和S-8等4种大孔吸附树脂对其除草活性物质的吸附性能,结果表明X-5树脂能够较好地吸附该除草活性物质.选用50%乙醇、95%乙醇、乙酸乙酯和丙酮4种溶剂为供试洗脱溶剂,采用2种洗脱方案,对其最佳洗脱溶剂进行了选择,生长抑制作用和种子萌发试验结果表明,以方案1洗脱为最佳.     

Inhibition of Rust Diseases of Cereals by Metabolic Products of Verticillium chlamydosporium

Verticillium chlamydosporium produced in submers culture several antifungal and/or phytotoxic compounds which were detected in a bioassay by using the pathogen-host system Puccinia coronata and oat seedlings. The antifungal compounds were also tested against P. recondita on wheat and P. sorghi on corn seedlings. The production of the active metabolic compounds highly depended on the nutrient solution (peptone-Czapek [PC] and malt extract [ME]) and on the fermentation times. Cell-free filtrates of PC-cultures of the fungus were highly phytotoxic; the fungitoxic and phytotoxic compounds were heat-labile and dialyzable. The ethyl acetate extracts of the PC-culture filtrates contained only the antifungal active substances. The antifungal compounds in ME-culture filtrates proved to be heat-stable, could be dialyzed and extracted with ethyl acetate. Ethyl acetate extracts of PC- and ME-culture filtrates at concentrations of 500 μg/ml reduced rust disease incidence by up to 80 % compared to the control treatment. Further studies with extracts of ME-culture filtrates displayed a distinct protective but no systemic activity. The extract interfered with the development of several infection structures of the rust fungi, mostly with the growth of germ tubes as well as with the formation of the aappressoria and haustorial mother cells. Three rust-active fractions were obtained by preparative layer chromatography on silica gel. One of these fractions exhibited phytotoxic activity. The most active antifungal fraction is identical with the macrolid antibiotic monorden which caused a desorientated spiral growth in P. coronata germlings on oat leaves.     

7-hydroxytropolone is the main metabolite responsible for the fungal antagonism of Pseudomonas donghuensis strain SVBP6

Pseudomonas donghuensis strain SVBP6, an isolate from an agricultural plot in Argentina, displays a broad-spectrum and diffusible antifungal activity, which requires a functional gacS gene but could not be ascribed yet to known secondary metabolites typical of Pseudomonas biocontrol species. Here, we report that Tn5 mutagenesis allowed the identification of a gene cluster involved in both the fungal antagonism and the production of a soluble tropolonoid compound. The ethyl acetate extract from culture supernatant showed a dose-dependent inhibitory effect against the phytopathogenic fungus Macrophomina phaseolina. The main compound present in the organic extract was identified by spectroscopic and X-ray analyses as 7-hydroxytropolone (7HT). Its structure and tautomerism was confirmed by preparing the two key derivatives 2,3-dimethoxy- and 2,7-dimethoxy-tropone. 7HT, but not 2,3- or 2,7-dimethoxy-tropone, mimicked the fungal inhibitory activity of the ethyl acetate extract from culture supernatant. The activity of 7HT, as well as its production, was barely affected by the presence of up to 50 μM added iron (Fe⁺²). To summarize, P. donghuensis SVBP6 produces 7HT under the positive control of the Gac-Rsm cascade and is the main active metabolite responsible for the broad-spectrum inhibition of different phytopathogenic fungi.     

Mitochondrial oxidative burst involved in apoptotic response in oats

Apoptotic cell response in oats is induced by victorin, a host-selective toxin secreted by Cochliobolus victoriae and thought to exert toxicity by inhibiting mitochondrial glycine decarboxylase (GDC) in Pc-2/Vb oats. We examined the role of mitochondria, especially the organelle-derived production of reactive oxygen species (ROS), in the induction of apoptotic cell death. Cytofluorimetric analysis showed that victorin caused mitochondrial deltaPsim breakdown and mitochondrial oxidative burst. Ultrastructural analysis using a cytochemical assay based on the reaction of H2O2 with CeCl3 detected H2O2 eruption at permeability transition pore-like sites on the mitochondrial membrane in oat cells treated with victorin. ROS generation preceded the apoptotic cell responses seen in chromatin condensation and DNA laddering. Both aminoacetonitrile (a specific GDC inhibitor) and antimycin A (a mitochondrial complex III inhibitor) also induced mitochondrial H2O2 eruption, and led to the apoptotic response in oat cells. ROS scavengers such as N-acetyl-l-cysteine and catalase suppressed the mitochondrial oxidative burst and delayed chromatin condensation and DNA laddering in the victorin- or antimycin A-treated leaves. These findings indicate possible involvement of mitochondria, especially mitochondrial-derived ROS generation, as an important regulator in controlling apoptotic cell death in oats.     

Oligolignans in the wood of <Emphasis Type="Italic">Picea obovata</Emphasis> Ledeb.

We studied the chemical composition of the phenolic complex and the structure of oligomeric lignans in Siberian spruce (Picea obovata Ledeb.). We used the wood of Siberian spruce collected near the city of Irkutsk. The extractives were isolated from ground wood (particle size: 10–15 mm; humidity: 5.9%) by three-stage acetone extraction. The extract was separated by consecutive treatment with the solvents with increasing polarity: hexane, ethyl acetate and n-butanol. The main amount of phenolic compounds (lignans) was concentrated in the ethyl acetate fraction and it was 0.7% in absolutely dry wood (adw).The ethyl acetate fraction of spruce wood extract was separated by silica gel column; a chloroform-acetone mixture was used as the eluent (the concentration of acetone in the mixture was increased from 0 to 100%). Monomeric lignans (~60–65% of the ethyl acetate fraction of spruce wood extract), oligomeric lignans (~20–25%), and polymer lignans (~12–15%) were isolated.We also obtained ¹³C nuclear magnetic resonance (NMR) spectra for the main monomeric, oligomeric and polymeric lignan compounds of phenolic complexes. It was found that oligomeric and polymeric fractions contain monomeric lignan units with the butyrolactone cycle, primarily, the fragments with hydroxymatairesinol structure. Oligomeric lignans contain the fragments with a pinoresinol and lariciresinol structure. All the monomeric structural units are characterized by the guaiacyl substitution type of the aromatic cycles.A preliminary study has been performed on the antiviral and antioxidant activity of the ethyl acetate fraction of acetone extract of Siberian spruce wood. It was found that the lignan complex is active against Coxsackie B4 virus in cell culture and in the pancreatitis model in white mice, reducing the activity of enteroviruses in the cell culture approximately 100 times. The antioxidant activity rate of polyphenol complex of Siberian spruce wood is comparable to that of a known antioxidant dihydroquercetin.     

Antibacterial activity of three South Indian seagrasses, <Emphasis Type="Italic">Cymodocea serrulata</Emphasis>, <Emphasis Type="Italic">Halophila ovalis</Emphasis> and <Emphasis Type="Italic">Zostera capensis</Emphasis>

The antibacterial properties of the three seagrasses namely Cymodocea serrulata, Halophila ovalis and Zostera capensis were tested against the human pathogens Staphylococcus aureus, Bacillus cereus, B. subtilis, Escherichia coli, Salmonella paratyphi, Salmonella typhimurium and Micrococcus luteus, using six different solvents namely, petroleum ether, chloroform, ethyl acetate, acetone, methanol and water. Ethyl acetate and methanol extracts showed maximum activity against most of the pathogens when compared to other solvents. Experiments are underway to isolate active compound(s) implicated in controlling the growth of the pathogens in vitro.     

Phosphatidic acid induces leaf cell death in Arabidopsis by activating the Rho-related small G protein GTPase-mediated pathway of reactive oxygen species generation

Phosphatidic acid (PA) level increases during various stress conditions. However, the physiological roles of this lipid in stress response remain largely unknown. In this study, we report that PA induced leaf cell death and elevated the levels of reactive oxygen species (ROS) in the whole leaf and single cells. To further elucidate the mechanism of PA-induced cell death, we then examined whether Rho-related small G protein (ROP) 2, which enhanced ROS production in an in vitro assay, is involved in PA-induced ROS production and cell death. In response to PA, transgenic leaves of Arabidopsis expressing a constitutively active rop2 mutant exhibited earlier cell death and higher levels of ROS than wild type (WT), whereas those expressing a dominant-negative rop2 mutant exhibited later cell death and lower ROS. However, in the absence of exogenous PA, no spontaneous cell death or elevated ROS was observed in constitutively active rop2 plants, suggesting that the activation of ROP GTPase alone is insufficient to activate the ROP-mediated ROS generation pathway. These results suggest that PA modulates an additional factor required for the active ROP-mediated ROS generation pathway. Therefore, PA may be an important regulator of ROP-regulated ROS generation and the cell death process during various stress and defense responses of plants.     

Repellency and toxicity of extract from Francoeria crispa (Forsk.) to Eutetranychus orientalis (Klein) (Acari: Tetranychidae)

Laboratory bioassays were conducted to characterise the activity of the extract Francoeria crispa (Forsk.) (Family: Compositae) against the citrus brown mite, Eutetranychus orientalis (Klein). Ethyl acetate was tested for preparing the crude extract of F. crispa. The extract was tested for its toxicity against eggs and adult females of the pest E. orientalis. Ethyl acetate extract of F. crispa affected the behaviour, toxicity and fecundity of females under laboratory conditions. The extract had similar toxic effects on egg stage and adult females of E. orientalis (LC₅₀ = 0.00050 g/ml), respectively. Leaf discs treated with increasing concentrations of ethyl acetate extract of F. crispa showed a high percentage of repellency (97.45%), respectively. Treated females with LC₅₀ concentration of ethyl acetate extract showed a higher remarkable percentage of mortality as well as a reduction in the total number of eggs laid during 7 days. Ten isolated fractions of ethyl acetate crude extract from F. crispa were detected. The results clearly indicate that the isolate number (10) was the most toxic isolate on eggs and females of E. orientalis (LC₅₀ = 0.00014 and 0.000125 g/ml), respectively.     

葎草不同溶剂提取物和萃取物的杀虫活性

对律草[Humulus scandens(Lour.) Merr.]全草干粉的不同溶剂提取物以及丙酮提取物的不同溶剂萃取物的杀虫活性进行了研究.在葎草的石油醚、苯、三氯甲烷、乙酸乙酯、丙酮和乙醇提取物(333 g·L-1)中,丙酮和乙醇提取物对小菜蛾(Plutella xylostella L.)的触杀作用较强,其中丙酮提取物的触杀作用最强,试虫的48h校正死亡率达到86.67％.6种溶剂提取物(100g·L-1)对小菜蛾的拒食活性均较弱,24和48 h拒食率仅为9.65％～ 20.45％.6种溶剂提取物对棉蚜(Aphis gossypii Glover)均有较强的触杀效果.其中丙酮提取物的触杀作用最强,经50g·L-1丙酮提取物处理后棉蚜24和48 h的校正死亡率分别为76.78％和85.64％,而经100g·L-1丙酮提取物处理后棉蚜24和48 h的校正死亡率分别为82.63％和92.53％.丙酮提取物的石油醚、三氯甲烷和乙酸乙酯萃取物对小菜蛾均有一定的触杀活性,且随萃取物浓度提高及处理时间的延长触杀活性增强;其中石油醚萃取物的触杀作用最强,经25.0g·L-1石油醚萃取物处理后小菜蛾24和48 h校正死亡率分别达到80.00％和96.67％ 研究结果表明:葎草丙酮提取物对小菜蛾和棉蚜的杀虫活性均最强,其主要有效杀虫活性成分存在于丙酮提取物的石油醚萃取物中.     

Bax translocation mediated mitochondrial apoptosis and caspase dependent photosensitizing effect of Ficus religiosa on cancer cells

The main aim of the present work was to investigate the potential effect of acetone extract of Ficus religosa leaf (FAE) in multiple apoptosis signalling in human breast cancer cells. FAE treatment significantly induced dose and time dependent, irreversible inhibition of breast cancer cell growth with moderate toxicity to normal breast epithelial cells. This observation was validated using Sulforhodamine B assay. Cell cycle analysis by Flow cytometry showed cell cycle arrest in G1 phase and induction of sub-G0 peak. FAE induced chromatin condensation and displayed an increase in apoptotic population in Annexin V-FITC/PI (Fluorescein isothiocyanate/Propidium iodide) double staining. FAE stimulated the loss of mitochondrial membrane potential in multiple breast cancer cell lines when compared to normal diploid cells. To understand the role of Bax in FAE induced apoptosis, we employed a sensitive cell based platform of MCF-7 cells expressing Bax-EGFP. Bax translocation to mitochondria was accompanied by the disruption of mitochondrial membrane potential and marked elevation in LEHDase activity (Caspase 9). Consistent with this data, FAE induced Caspase activation as evidenced by ratio change in FRET Caspase sensor expressing MCF-7 cell line and cleavage of prominent Caspases and PARP. Interestingly, FAE accelerated cell death in a mitochondrial dependent manner in continuous live cell imaging mode indicating its possible photosensitizing effect. Intracellular generation of reactive oxygen species (ROS) by FAE played a critical role in mediating apoptotic cell death and photosensitizing activity. FAE induced dose and time dependent inhibition of cancer cell growth which was associated with Bax translocation and mitochondria mediated apoptosis with the activation of Caspase 9 dependent Caspase cascade. FAE also possessed strong photosensitizing effect on cancer cell line that was mediated through rapid mitochondrial transmembrane potential loss and partial Caspase activation involving generation of intracellular ROS.     

Immunomodulatory Effect of Gymnema sylvestre (R.Br.) Leaf Extract: An In Vitro Study in Rat Model

Gymnema sylvestre Wild R.Br (family: Asclepidaceae) is a valuable medicinal plant used in folk medicine to treat diabetes, obesity, asthma etc. in India for antiquity. Diabetes mellitus is a syndrome characterized immunologically by lymphocyte apoptosis and reduced cell-mediated and humoral immunity. Modulation of immune responses to alleviate diseases has been of interest, and traditional herbal medicines may play an important role in this regard. In this study, we aim to evaluate the immunomodulatory potential of methanolic extract of G. sylvestre leaf using rat model. HPLC analysis of leaf extract was carried out for gymnemic acid. The method involves the initial hydrolysis of gymnemic acids, the active ingredients, to a common aglycone followed by the quantitative estimation of gymnemagenin, using gymnemagenin as reference standard. Gymnemic acid content was 2.40% (w/w) in G. sylvestre leaf extract. In vitro immunomodulatory activity of the methanolic extract of G. sylvestre leaf (1–200μg/ml) was evaluated by gauging its effects on nitroblue tetrazolium reduction and nitrite release in rat peritoneal macrophages and on mitogen (ConA, PHA and LPS) induced splenic lymphocyte proliferation. G. sylvestre leaf extract showed significant (<0.05) enhancement in NO and ROS generation in macrophages and in proliferation of lymphocytes in dose dependent manner. EC₅₀ value was 3.10, 3.75 and 2.68μg/ml for NBT reduction, nitrite release and lymphoproliferation, respectively. Potential effect was observed at 100 μg/ml in NO and ROS generation in macrophages and 20 μg/ml in lymphocyte proliferation. G. sylvestre leaf extract stimulates macrophage reactivity, increasing the level of activity even higher when combined with PMA or LPS. These findings suggest the presence of active compounds, gymnemic acid, in methanolic extract of G. sylvestre leaf that stimulates both myeloid and lymphoid components of immune system, and therefore can restore the innate immune function. Through this study, the traditional knowledge of anti-diabetic property of G. sylvestre is scientifically supplemented with its immunomodulatory properties.     

H2O2-induced Leaf Cell Death and the Crosstalk of Reactive Nitric/Oxygen Species([F])

In plants, the chloroplast is the main reactive oxygen species (ROS) producing site under high light stress. Catalase (CAT), which decomposes hydrogen peroxide (H₂O₂), is one of the controlling enzymes that maintains leaf redox homeostasis. The catalase mutants with reduced leaf catalase activity from different plant species exhibit an H₂O₂‐induced leaf cell death phenotype. This phenotype was differently affected by light intensity or photoperiod, which may be caused by plant species, leaf redox status or growth conditions. In the rice CAT mutant nitric oxide excess 1 (noe1), higher H₂O₂ levels induced the generation of nitric oxide (NO) and higher S‐nitrosothiol (SNO) levels, suggesting that NO acts as an important endogenous mediator in H₂O₂‐induced leaf cell death. As a free radical, NO could also react with other intracellular and extracellular targets and form a series of related molecules, collectively called reactive nitrogen species (RNS). Recent studies have revealed that both RNS and ROS are important partners in plant leaf cell death. Here, we summarize the recent progress on H₂O₂‐induced leaf cell death and the crosstalk of RNS and ROS signals in the plant hypersensitive response (HR), leaf senescence, and other forms of leaf cell death triggered by diverse environmental conditions. [ Chengcai Chu (Corresponding author)]     

Biological activity of maize leaf extracts against the African armyworm,Spodoptera exempta

Maize (Zea mays L.) leaf tissue of cv Bastille and cv Michoacan 12 was extracted with n-hexane. The extracts were bioassayed against 5th instar African armyworm,Spodoptera exempta (Walker)(Lepidoptera: Noctuidae), by feeding the larvae on agar based media or sucrose impregnated glass fibre discs. The hexane extract of the ‘resistant’ cv Bastille exhibited feeding deterrency and toxicity which were not shown by the ‘susceptible’ cv Michoacan 12. The hexane extract of cv Bastille was adsorbed onto silica gel, the solution filtered off and the adsorbed component taken up into ethyl acetate. Bioassay of these fractions indicated that the toxic and deterrent action was retained in the ethyl acetate fraction. Preparative thin layer chromatography of the ethyl acetate fraction isolated two biologically active constituents. These were both growth inhibitors and lethal by ingestion to the 5th instar African armyworm. Implications for resistance in maize varieties to insect pests are discussed.     

Studies on Enzymatic Synthesis of Oligosaccharides

The overproduction of tumor necrosis factor-α (TNF-α) was suppressed by orally administering a perilla leaf extract (PLE). When mice were successively injected with OK-432, severe TNF-α was induced in the serum, but this elevated TNF-α level was reduced after an oral administration of PLE (400 μl/mouse). Oral administration of PLE also inhibited TNF-α production that was induced by muramyl dipeptide (500 μg/mouse) and OK-432 (3 KE/mouse). These characteristics were obtained from all strains of perilla. The inhibitory activity against TNF-α production was heat-stable, and the existence of several active molecules was suggested. When PLE was passed through an ultrafilter, the inhibitory activity against TNF-α production was collected in those fractions with a mass of 0.5 to 1 kDa and more than 10 kDa. When PLE was solvent-extracted, the strongest activity was recognized with aqueous preparation, although significant activity was also detected in preparations extracted with n-hexane and ethyl acetate. These findings suggest that the daily use of certain functional foods may be useful for controlling the host defense system.     

Red Mexican grapefruit: a novel source for bioactive limonoids and their antioxidant activity

Citrus limonoids have shown to inhibit the growth of cancer in colon, lung, mouth, stomach and breast in animal and cell culture studies. For the first time in the present study, an attempt has been made to isolate antioxidant fractions and five limonoids from red Mexican grapefruit seeds. Defatted seed powder was successively extracted with hexane, ethyl acetate (EtOAc), acetone, methanol (MeOH) and MeOH/water and the extracts were concentrated under vacuum. Radical scavenging activity of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and total phenolic content were also measured for comparison with the antioxidant capacity in the phosphomolybdenum method for the above extracts. Acetone and MeOH extracts, respectively, showed the highest (85.7%) and lowest (53.3%) radical scavenging activity, at 500 ppm. The total phenolic contents were found to be highest in the acetone extract (15.94%) followed by the MeOH extract (5.92%), ethyl acetate extract (5.54%) and water extract (5.26%). Antioxidant capacity of the extracts as equivalents to ascorbic acid (micromol/g of the extract) was in the order, EtOAc extract > acetone extract > water extract > methanol extract. Furthermore, the EtOAC and acetone extracts were loaded onto silica gel columns to obtain four limonoid aglycons. MeOH fraction was loaded onto a dowex-50 and sepabeads resin column to obtain a limonoid glucoside. The purity of the isolated five compounds was analyzed by HPLC using a C18 column and UV detection at 210 nm. Finally, the structures of the compounds were identified as obacunone, nomilin, limonin, deacetylnomilin (DAN) and limonin-17-beta-D-glucopyranoside (LG) using 1H and 13C NMR studies.     

Evidence for Naturally Occurring Inhibitors of Archegonial Differentiation in Lygodium japonicum

Archegonial differentiation in prothallia of Lygodium japonicum was inhibited when the filtrate of conditioned medium or the extracts of prothallia with organic solvents were added to the medium. By varying the timing of treatment with the methanol extract, archegonial differentiation was shown to start at least 4 days before microscopically detectable change. The inhibitory effect of methanol extract was nullified by transferring the treated plants to a fresh medium omitting the methanol extract, so that the archegonial formation became discernible 6 days after the transfer. The inhibitory activity was stable in both acidic and basic solutions at room temperature, and was partially lost by boiling at pH 3 or 11 for 30 min. The inhibitor, which could be retrieved from the filtrate and the methanol extract, was fractionated into the neutral ethyl acetate fraction, but was not found in the acidic ethyl acetate fraction and in the aqueous residue. At least two active zones were separated on thin layer chromatograms of the ethyl acetate extracts from the filtrate and the methanol extract, and the relative flow-rates of each active zone from these two sources were very similar. The evidence described above indicates that specific inhibitors of archegonial differentiation may be produced in the tissue of prothallia of Lygodium and eventually be secreted to the medium.     

Inhibitory effect of Torilis anthriscus on growth of microorganisms

Antibacterial and antifungal activities of aqueous, ethanol and ethyl acetate extract of Torilis anthriscus (L.) Gmel. (Apiaceae) were tested in vitro against ten species of bacteria and five of fungi. Antimicrobial properties were determined by disk diffusion and broth tube dilution method. In the minimum inhibitory concentrations (MICs), the ethanol extract showed the highest activity, followed by the ethyl acetate extract and the aqueous extract against bacterial species, while the extracts were inactive against the tested fungi species. The most active extract was chosen to examine the effects of its combinations with commercial antibiotics by checkerboard method. The obtained results showed that the interactions between ethanol extract/streptomycin and ethanol extract/chloramphenicol were additive and indifferent against the tested human-pathogenic bacteria. Synergism and antagonism were not observed.     

Bioefficacy of Aristolochia tagala Cham. against Spodoptera litura Fab. (Lepidoptera: Noctuidae)

Bioefficacy of leaf and root extracts of Aristolochia tagala Cham. at different concentrations was evaluated at room temperature against Spodoptera litura Fab. Effects on feeding, larvicidal and pupicidal activities and larval–pupal duration were studied. Higher antifeedant activity (56.06%), lethal concentration for feeding inhibition (3.69%), larvicidal (40.66%), pupicidal (28%), total mortality (68.66%) and prolonged larval–pupal duration (12.04–13.08 days) were observed in ethyl acetate leaf extract at 5.0% concentration. Dose dependant effect of test extracts was observed. This plant could be used to isolate active principles and to develop a new botanical formulation in pest management programmes.