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Toxicity and receptor binding properties of Bacillus thuringiensis delta-endotoxins to the midgut brush border membrane vesicles of the rice leaf folders, Cnaphalocrocis medinalis and Marasmia patnalis.

Pesticidal activity and receptor-binding properties of Bacillus thuringiensis toxins to rice leaf folders, Cnaphalocrocis medinalis and Marasmia patnalis, were investigated. Saturation and competition binding experiments were done with iodine ((125)1)-labeled Bt proteins and brush border membrane vesicles prepared from the midgut of C. medinalis and M. patnalis. The results show saturable, specific, and high-affinity binding of all toxins except Cry2A toxin. Cry1Aa and Cry2A toxins were bound with low affinity but with high binding site concentration. Heterologous competition experiments showed that Cry1Aa, Cry1Ab, and Cry1Ac recognized or shared the same binding site that is different from the binding site for Cry2A toxin. Iodine ((125)I)-labeled Cry1Ac and Cry1Ab toxins were used in ligand blot experiments to detect specific binding proteins in brush border membrane vesicles of C. medinalis and M. patnalis. Cry1Ab toxin protein binds to 205-kDa and 200-kDa proteins respectively in case of C. medinalis and M. patnalis. The apparent molecular mass of the protein bound to labeled Cry1Ac toxins was identified as a 120-kDa protein in both C. medinalis and M. patnalis.

Animals↗

Cytotoxicity of fungal metabolites to lepidopteran (Spodoptera frugiperda) cell line (SF-9).

The toxicity of sixteen fungal metabolites produced by some entomopathogenic fungi or biological control fungi agents was evaluated on lepidopteran Spodoptera frugiperda (SF-9) cell line by Trypan blue dye exclusion and MTT-colorimetric assay, after 48 h of incubation. No statistical difference was found between IC50values (50% Inhibiting Concentration) and CC50 values (50% Cytotoxicity Concentration) obtained by MTT test and Trypan blue dye exclusion for each fungal metabolite. By MTT assay, the cytotoxicity ranking was fusarenon X (IC50 0.3 microM) = diacetoxyscirpenol (IC50 0.5 microM) = beauvericin (IC50 2.5 microM) = nivalenol (IC50 5.3 microM) = enniatin (IC50 6.6 microM) > or = gliotoxin (IC50 7.5 microM) > zearalenone (IC50 17.5 microM) > deoxynivalenol (IC50 47.6 microM). By Trypan blue dye exclusion the cytotoxicity ranking was fusarenon X (CC50 0.4 microM) = diacetoxyscirpenol (CC50 1.1 microM) beauvericin = (CC50 3.0 microM)=gliotoxin (CC50 4.0 microM) = enniatin (CC50 6.7 microM) > or = nivalenol (CC50 9.5 microM) > zearalenone (CC50 18.3 microM) > deoxynivalenol (CC50 45.0 microM). The comparison with other bioassays showed that the SF-9 insect cell line could represent a further tool to screen for the toxic effects of fungal metabolites especially for beauvericin, gliotoxin, and zearalenone.

Animals↗

Effect of temperature and host plant leaf morphology on the efficacy of two entomopathogenic biocontrol agents of Thrips palmi (Thysanoptera: Thripidae).

The efficacy of two entomopathogenic biocontrol agents, Steinernema feltiae (Filipjev) and Verticillium lecanii (Zimmerman) Viégas (reclassified now as Lecanicillium muscarium (Petch) Zare & Gams), against Thrips palmi Karny was investigated. Assessments of the effect of temperature on the efficacy of S. feltiae indicated that higher mortality of T. palmi was recorded at 20 degrees C compared to either 15 or 25 degrees C, whereas significantly higher T. palmi mortality followed application of L. muscarium at 25 degrees C. Testing the control agents efficacy on three host plants; chrysanthemum, sweet pepper and cucumber, under constant temperature and high humidity conditions produced no significant difference in the level of T. palmi larval mortality on each host plant. Incorporating the chemical insecticide imidacloprid with both biological agents in a combined control strategy increased T. palmi juvenile mortality. The potential role of S. feltiae and L. muscarium within integrated pest management programmes for the control of T. palmi is discussed.

Animals↗

Toxicity and repellency of hot pepper extracts to spider mite, Tetranychus urticae Koch.

Increasing concern about persistence and environmental impact of synthetic pesticide residues require development of biodegradable and environmentally safe alternatives. The potential of using fruit extracts of hot pepper as alternatives to synthetic acaricides for controlling the two-spotted spider mite, Tetranychus urticae Koch, is explored in this study. Twenty-four Capsicum accessions (Solanaceae) were screened for their toxicity and repellency to the spider mites. Crude extracts from fruits of C. chinense, C. frutescens, C. baccatum, C. annuum, and C. pubescens were prepared in methanol and tested for their acaricidal properties. Spider mite mortality was greatest (45%) when fruit extract of accession Grif-9169 (C. annuum) was used. Results from diving board bioassays indicated that mites avoided filter paper strips treated with hot pepper extracts from accessions PI-596057 (C. baccatum), PI-195299 (C. annuum), and Grif- 9270 (C. annuum). This investigation suggests that methanolic extracts of these three accessions may have a great potential for repelling spider mites and should be field-tested on a large-scale to assess their value in managing populations of spider mites, which could reduce reliance on synthetic acaricides. An attempt was made to correlate repellency with chemical constituents of fruit extracts of the most repellent accessions to identify chemical sources of repellency. Capsaicin and dihydrocapsaicin, the pungent components of pepper fruit, were not correlated with toxicity or repellency, indicating that these are not likely related to the toxicity or repellency of the pepper fruit extracts. Other, unidentified chemicals are likely responsible for toxicity and repellency to the two-spotted spider mite.

Animals↗

Effect of drying period and soil moisture on egg hatch of the tadpole shrimp (Notostraca: Triopsidae).

Tadpole shrimp (Triops spp.) are potential biological control agents against larval mosquitoes in temporary ponds and flood-irrigated fields. In some rice field situations, however, they may become pests that uproot and eat young rice plants. In cursory observations, it has been reported that tadpole shrimp eggs do not readily hatch on flooding when the soil or substrate containing eggs is moist before flooding. The relationship between drying (moisture content) of soil and tadpole shrimp hatch was determined in studies conducted in mesocosms at the University of California Aquatic and Vector Control Research facilities at Riverside and at Oasis in the Coachella Valley of southern California. In laboratory hatching trials, an increase in hatch of Triops longicaudatus (LeConte) with declining soil moisture content was demonstrated (t = 8.4, P less than 0.001; r2 = 0.76). In field trials in mesocosms at Riverside, egg hatch increased with increased drying period and declining soil moisture content (G = 29.8, P less than 0.01). No hatch of eggs occurred after 3 d of drying, when soil moisture content was high, but a high level of hatching occurred after 7 and 14 d of drying, when soil moisture declined to low levels. At Oasis, soil moisture did not decrease with drying time because of porous soil and capillary action of water from adjacent flooded mesocosms and thick vegetation covering the pond bottoms. Therefore, hatch rates at Oasis were not associated with the length of the drying period (G = 35, P greater than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Infection of malaria (Anopheles gambiae s.s.) and filariasis (Culex quinquefasciatus) vectors with the entomopathogenic fungus Metarhizium anisopliae.

BACKGROUND: Current intra-domiciliary vector control depends on the application of residual insecticides and/or repellents. Although biological control agents have been developed against aquatic mosquito stages, none are available for adults. Following successful use of an entomopathogenic fungus against tsetse flies (Diptera: Glossinidae) we investigated the potency of this fungus as a biological control agent for adult malaria and filariasis vector mosquitoes. METHODS: In the laboratory, both sexes of Anopheles gambiae sensu stricto and Culex quinquefasciatus were passively contaminated with dry conidia of Metarhizium anisopliae. Pathogenicity of this fungus for An. gambiae was further tested for varying exposure times and different doses of oil-formulated conidia. RESULTS: Comparison of Gompertz survival curves and LT50 values for treated and untreated specimens showed that, for both species, infected mosquitoes died significantly earlier (p < 0.0001) than uninfected control groups. No differences in LT50 values were found for different exposure times (24, 48 hrs or continuous exposure) of An. gambiae to dry conidia. Exposure to oil-formulated conidia (doses ranging from 1.6 x 10(7) to 1.6 x 10(10) conidia/m2) gave LT50 values of 9.69 +/- 1.24 (lowest dose) to 5.89 +/- 0.35 days (highest dose), with infection percentages ranging from 4.4-83.7%. CONCLUSION: Our study marks the first to use an entomopathogenic fungus against adult Afrotropical disease vectors. Given its high pathogenicity for both adult Anopheles and Culex mosquitoes we recommend development of novel targeted indoor application methods for the control of endophagic host-seeking females.

Age Factors↗

Cytolytic toxin Cyt1Aa of Bacillus thuringiensis synergizes the mosquitocidal toxin Mtx1 of Bacillus sphaericus.

Using the shuttle vector pBU4, the mosquitocidal toxin gene mtx1 from Bacillus sphaericus strain SSII-1 was introduced into an acrystalliferous strain of B. thuringiensis both individually and in combination with the accessory protein gene p20 and the cytolytic protein gene cyt1Aa from B. thuringiensis subsp. israelensis. Bioassay results indicated that the recombinants B-pMT4(Mtx1) and B-pMT9(Mtx1), both individually containing mtx1, had moderate toxicities to binary toxin susceptible and binary toxin resistant Culex quinquefasciatus larvae during the vegetative growth stage, but that their toxicities declined rapidly during the sporulation phase. The LC50 values were 2.5 and 4.8 mg/ml respectively, against 3-4 instar susceptible and resistant larvae for the final sporulated cultures of recombinants B-pMT9(Mtx1), and little toxicity was detected for B-pMT4(Mtx1). Meanwhile, the recombinant B-pMPX2(Mtx1+Cyt1Aa) expressing Mtx1, P20 alone, and Cyt1Aa in combination had stable toxicities during both the vegetative phase and the sporulation phase, with a LC50 ranging from 0.45-0.58 mg/ml. Furthermore, expression of Cyt1Aa appeared to enhance the activity of Mtx1 to target mosquito larvae, suggesting a synergism between Cyt1Aa and Mtx1 toxins.

Animals↗

Streptomyces avermitilis from marine.

The insecticidal strain 173 was isolated from marine source and its activity was explored by the bioassay of brine shrimp and Helicoverpa armigera. Based on morphological, physiological and molecular properties, the insecticidal strain 173 was identified as Streptomycetes avermitilis, which is the best insecticidal microorganism found in the terrestrial environment. The taxonomy of the strain 173, insecticidal spectrum and properties of the corresponding insecticidal antibiotics are reported.

Acetates↗

Synergistic interaction between Beauveria bassiana- and Bacillus thuringiensis tenebrionis-based biopesticides applied against field populations of Colorado potato beetle larvae.

Commercial biopesticides based on the fungal pathogen Beauveria bassiana strain GHA and the bacterial pathogen Bacillus thuringiensis tenebrionis were applied alone and in combination (tank mixed) against larval populations of the Colorado potato beetle, Leptinotarsa decemlineata, in small plots of potatoes over three field seasons. Interactions between the two products were evaluated in terms of pest-control efficacy. B. bassiana (formulated as Mycotrol) was applied at low and medium label rates of 1.25 and 2.5 x 10(13) conidia/ha, and B. thuringienis (formulated as Novodor) was applied at low and high label rates of 40.3 and 120.8 x 10(6) Leptinotarsa units/ha. Two weekly applications of the bacterial pesticide alone provided 50-85% control of beetle larvae within 14 days after the initial application, while applications of the mycopesticide alone produced no greater than 25% control. Maximum control, in nearly all tests, was produced by the combination of the two products. The combined treatments produced a statistically significant 6-35% greater reduction in larval populations than would have been predicted had the two biopesticides acted independently. This low-level synergistic interaction was observed during all field seasons and resulted from combinations at all rates, including, in one of two tests, the low rates of each product. These results indicate that B. thuringiensis and B. bassiana have strong potential for integrated biologically based management of Colorado potato beetle.

Animals↗

Pathogenicity of the fungus, Verticillium lecanii, to the green peach aphid, Myzus persicae (Hom.: Aphididae).

Pathogenicity of the hyphomycete, Verticillium lecanii (DAOM 198499), was investigated to aphid, Myzus persicae under laboratory conditions. Analysis of lethal effect of six various concentrations 0, 10(4), 10(5), 10(6), 10(7) and 10(8) conidia/ml of V. lecanii against third nymphal stage of M. persicae, indicated significant mortality on aphids. Mean comparisons showed that there was significant difference between treatments. Three days after treatment, aphid mortality observed and after 12 days minimal mortality was 17.77 in control and maximum was 100 percent related to 10(7) and 10(8) conidia/ml. LC50 and LT50 values were estimated by probit analysis and life test. LC50 value for aphid mortality was 1.4 x 10(4) conidia/ml, LT50 values for concentrations 10(4), 10(6), 10(6), 10(7) and 10(8) conidla/ml was 10, 10, 9, 8 and 6 days, respectively. In this experiment, net reproduction rate of aphid's (R0) decreased significantly when concentration increased. These observations showed that V. lecanii (DAOM 198499) can be an active biological agent against aphids.

Animals↗

The effect of chrysanthemum leaf trichome density and prey spatial distribution on predation of Tetranychus urticae (Acari: Tetranychidae) by Phytoseiulus persimilis (Acari: Phytoseiidae).

The effect of plant architecture, in terms of leaf hairiness, and prey spatial arrangement, on predation rate of eggs of the spider mite, Tetranychus urticae Koch, by the predatory mite Phytoseiulus persimilis Athias-Henriot was examined on cut stems of chrysanthemums. Three levels of leaf hairiness (trichome density) were obtained using two different chrysanthemum cultivars and two ages within one of the cultivars. The number of prey consumed by P. persimilis was inversely related to trichome density. At low prey densities (less than ten eggs per stem), prey consumption did not differ in a biologically meaningful way between treatments. The effect of prey spatial arrangement on the predation rate of P. persimilis was also examined. Predation rates were higher in prey patches on leaves adjacent to the release point of P. persimilis, but significantly greater numbers of prey were consumed in higher density prey patches compared to low density patches. The predators exhibited non-random searching behaviour, spending more time on leaves closest to the release point. The implications of these findings for biological control and predator-prey dynamics are discussed.

Animals↗

Using chimeric hypoviruses to fine-tune the interaction between a pathogenic fungus and its plant host.

Infectious cDNA clones of mild (CHV1-Euro7) and severe (CHV1-EP713) hypovirus strains responsible for virulence attenuation (hypovirulence) of the chestnut blight fungus Cryphonectria parasitica were used to construct viable chimeric viruses. Differences in virus-mediated alterations of fungal colony morphology, growth rate, and canker morphology were mapped to a region of open reading frame B extending from nucleotides 2,363 to 9, 904. By swapping domains within this region, it was possible to generate chimeric hypovirus-infected C. parasitica isolates that exhibited a spectrum of defined colony and canker morphologies. Several severe strain traits were observed to be dominant. It was also possible to uncouple the severe strain traits of small canker size and suppression of asexual sporulation. For example, fungal isolates infected with a chimera containing nucleotides 2363 through 5310 from CHV1-Euro7 in a CHV1-713 background formed small cankers that were similar in size to that caused by CHV1-EP713-infected isolates but with the capacity for producing asexual spores at levels approaching that observed for fungal isolates infected with the mild strain. These results demonstrate that hypoviruses can be engineered to fine-tune the interaction between a pathogenic fungus and its plant host. The identification of specific hypovirus domains that differentially contribute to canker morphology and sporulation levels also provides considerable utility for continuing efforts to enhance biological control potential by balancing hypovirulence and ecological fitness.

Ascomycota↗

Influence of disease-suppressive strains of Streptomyces on the native Streptomyces community in soil as determined by the analysis of cellular fatty acids.

Analysis of cellular fatty acid profiles was used to distinguish among introduced pathogen- suppressive strains and indigenous strains of Streptomyces spp. isolated from soil of field plots established to test the efficacy of Streptomyces strains PonSSII and PonR in the biological control of potato scab. Reference libraries of fatty acid profiles were developed for a collection of known pathogenic strains and the introduced suppressive strains. Population densities of pathogen-related, suppressive, and saprophytic Streptomyces strains were determined from the relationship of field isolates to mean library profiles using cluster analysis and the unweighted pair-group method using arithmetic averages. Community diversity was similarly determined. Streptomyces strains PonSSII and PonR were distinguished from each other and from the pathogen group (which clustered together) based on fatty acid profiles. The introduced, suppressive strains successfully colonized the soil and represented 2-19% of the isolates sampled over 2 years. The introduction of the suppressive strains inhibited the population of strains related to the pathogen library at each sample date; the pathogen population was substantially lower in soil from treatments where the suppressive strains were introduced compared with the nonamended control. At harvest, the pathogen-related population was suppressed 85-93 and 36-44% in 1991 and 1992, respectively, in treatments with the suppressive strains compared with the nonamended control. Diversity of the community was not affected by the introduced strains, and diversity and equitability indices were similar among treatments at any sample time. The inhibition of the pathogen-related population was correlated with a reduction of scab symptoms observed in the field plots into which the suppressive strains were introduced. Implications of a fundamental shift in the pathogen-related population in response to the introduction of the suppressive strains for long-term biological control of potato scab are encouraging.

Fatty Acids↗

Serratia entomophila bacteriophages: host range determination and preliminary characterization.

Eight bacteriophages specific to Serratia entomophila, a commercially available bacterial pathogen of the New Zealand grass grub (Costelytra zealandica), were characterized by host range determination, morphology and restriction endonuclease patterns of DNA. Phages were originally isolated from grass grub larvae and fermenter broth where phages had disrupted large-scale production of S. entomophila. Seven of the phages (CW1-CW5, BC, and BT) had heads similar in size (approximately 60 x 60 nm) and long noncontractile tails (185 x 10 nm). Phage AgRP8 (P8) had a smaller head and a short tail structure. Restriction endonuclease analysis divided the phages into four groups: CW2, CW4, CW5, BC, and BT gave identical patterns, white CW1, CW3, and P8 each gave different patterns. Six distinct phage groups were distinguished by host range determination, after screening phages against 70 bacterial isolates: CW1, CW2/CW4, CW3, CW5, BC/BT, and P8. While confirming the indicated groupings by DNA analysis, it was possible to distinguish between some of the phages in the largest group: CW2/4 could be distinguished from CW5 and BC/BT. Screening of soil bacterial isolates of S. entomophila against nondiluted phages will aid in monitoring the establishment and persistence of strains applied for biological control of the grass grub.

Bacteriophages↗

[Evaluation of Trichoderma spp. as antagonist of Rhizoctonia solani in vitro and as biocontrol of greenhouse tomato plants].

Five Trichoderma isolates were compared in their ability for controlling Rhizoctonia solani attack to tomato plants in greenhouse and as antagonists of this pathogen in three independent laboratory assays. Four out of five isolates showed biocontrol ability and decreased pathogen growth and survival of its sclerotia in soil. Results suggest that dual cultures in Petri dishes and mycoparasitism assays against R. solani sclerotia may be useful for detecting isolates effective as biological control agents against this pathogen in tomato plants.

Agriculture↗