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Assessment of toxic potential of local Jordanian Bacillus thuringiensis strains on Drosophila melanogaster and Culex sp. (Diptera).

AIMS: To assess the toxic potential of different local Jordanian Bacillus thuringiensis isolates on larvae of Drosophila melanogaster and Culex sp. METHODS AND RESULTS: Scanning electron microscopy revealed the presence of spherical, bi-pyramidal, and bi-pyramidal and cuboidal parasporal bodies produced by the toxic isolates. Spherical inclusions dominated. The toxicity of the isolates to the two insects, determined using 24-well plates or vials, indicated that the 50% lethal concentration (LC50) of the bacterial suspension for D. melanogaster and Culex sp. larvae varied from 4.60 to 8.65, and from 5.30 to 6.74, respectively. CONCLUSION: Comparison of the LC50 values of isolate 82 with those of the reference strain B. t. israelensis showed that this isolate has a higher toxicity potential. SIGNIFICANCE AND IMPACT OF THE STUDY: Some local Jordanian B. thuringiensis isolates exhibit toxic potential that could be used to control some important pests, and could replace chemical pesticides.

Animals↗

The gut bacteria of insects: nonpathogenic interactions.

The diversity of the Insecta is reflected in the large and varied microbial communities inhabiting the gut. Studies, particularly with termites and cockroaches, have focused on the nutritional contributions of gut bacteria in insects living on suboptimal diets. The indigenous gut bacteria, however, also play a role in withstanding the colonization of the gut by non-indigenous species including pathogens. Gut bacterial consortia adapt by the transfer of plasmids and transconjugation between bacterial strains, and some insect species provide ideal conditions for bacterial conjugation, which suggests that the gut is a "hot spot" for gene transfer. Genomic analysis provides new avenues for the study of the gut microbial community and will reveal the molecular foundations of the relationships between the insect and its microbiome. In this review the intestinal bacteria is discussed in the context of developing our understanding of symbiotic relationships, of multitrophic interactions between insects and plant or animal host, and in developing new strategies for controlling insect pests.

Animals↗

Pathogenic action of Penicillium species on mosquito vectors of human tropical diseases.

In vitro bioassays were performed in order to assess the pathogenicity of 13 Penicillium strains in 2nd stage larvae of Aedes aegypti, Aedes fluviatilis, Anopheles aquasalis and Culex quinquefasciatus. Mortality rates began in the first 24 hours, ranging from 0 to 100%. P. corylophilum, P. fellutanum, P. implicatum, P. janthinellum, P. viridicatum and P. waksmanii species tested on Aedes aegypti larvae and at different concentrations presented mortality rates from 0 to 6.6%. These species, when tested on Aedes fluviatilis, Anopheles aquasalis and Culex quinquefasciatus larvae, presented a mortality rate of 3.33% to 100%. Susceptibility of Aedes fluviatilis, Aedes aegypti, Anopheles aquasalis and Culex quinquefasciatus mosquitoes to the tested agents, turn P. corylophilum and P. janthinellum into candidates for potential use in biological control of vectors.

Animals↗

A transgene-based, embryo-specific lethality system for insect pest management.

Biological approaches to insect pest management offer alternatives to pesticidal control. In area-wide control programs that cover entire regions, the sterile insect technique (SIT) can be used to successfully suppress economically important pest species by the mass release of sterilized pest organisms. However, conventional sterilization by ionizing radiation reduces insect fitness, which can result in reduced competitiveness of the sterilized insects. Here we report a transgene-based, dominant embryonic lethality system that allows for generation of large quantities of competitive but sterile insects without the need of irradiation. The system involves the ectopic expression of a hyperactive pro-apoptotic gene that causes embryo-specific lethality when driven by the tetracycline-controlled transactivator (tTA) under the regulation of a cellularization gene enhancer-promoter. We have successfully tested this system in Drosophila melanogaster. The embryonic lethality can be suppressed maternally, which will allow it to be combined with transgenic female-specific lethality systems to raise only vigorous but sterile males.

Animals↗

IgE-sensitization to predatory mites and respiratory symptoms in Swedish greenhouse workers.

BACKGROUND: Predatory mites are used as biological pesticides worldwide for control of spider mites and other pests in greenhouses. The aim of this study was to evaluate the impact of occupational exposure to Phytoseiulus persimilis and Hypoaspis miles on IgE sensitization among a large group of Swedish greenhouse workers and to examine the relationship between exposure and allergic asthma and rhinoconjunctivitis. METHODS: A total of 96 greenhouse workers from the southern part of Sweden, who were using the predatory mites for control of pests, were investigated with a questionnaire and a medical examination including lung function test. Blood samples were taken to test for allergen-specific IgE antibodies to Phytoseiulus persimilis and Hypoaspis miles as well as to Tetranychus urticae, Dermatophagoides pteronyssinus/farinae and Tyrophagus putrescentiae. RESULTS: Seventeen of the 96 workers were positive in ImmunoCAP to predatory mites: 17 to P. persimilis (17.7%) and 14 to H. miles (14.6%). Subjects sensitized to predatory mites were significantly more often atopic (13/17), defined as a positive Phadiatop, than those who lacked IgE against these mite species (17/79) (P <0.01). IgE antibodies to the red spider mite T. urticae were present among 23 subjects. Thirty-five of the investigated subjects displayed a positive ImmunoCAP to at least one of the investigated mite species. Furthermore, sensitization to any of the mites tested was significantly associated with asthma (OR=9.3) and rhinoconjunctivitis (OR=4.3). CONCLUSIONS: IgE sensitization to predatory mites, P. persimilis and H. miles, is common among greenhouse workers. The findings stress the importance of improved allergen avoidance in greenhouse environments.

Adult↗

Biocontrol agents of Botrytis cinerea tested in climate chambers by making artificial infection on tomato leafs.

To reduce the use of chemical agents, that are causing damage to the environment, in the fight against Botrytis cinerea, different BCA's were tested for their possibility to control Botrytis cinerea in a biological way. In order to investigate the effectiveness of the different micro organisms and Elot-Vis, experiments were carried out in climate chambers with 5 weeks old tomato plants. Leafs on the plant were inoculated with drops of a suspension that contained spores of Botrytis cinerea. The possible antagonists that were tested in these experiments were Trichoderma harzianum (Trichodex), T. asperellum (Biofungus), T. hamatum (T382), Bacillus subtilis (Serenade and Phytovit) and Pseudomonas aeruginosa (7NSK2 and KMPCH). For all these different micro organisms the direct and the indirect influence on Botrytis cinerea was investigated. In tests where the direct influence of the antagonists was examined, the spores of the moulds or the bacteria were suspended together with spores of Botrytis cinerea and subsequently drops of this suspension were pippeted on the leafs. After a while by ideal circumstances for Botrytis cinerea the infections on the inoculated leafs were counted. For the indirect influence of the antagonists, also leafs of 5 weeks old tomato plants were inoculated with a suspension of Botrytis cinerea spores. The roots of the tomato plants that were used for testing the indirect influence were treated during there growth with a suspension of the antagonist to see if induced systemic resistance pathway (ISR) was activated. For testing the effectiveness of Elot-Vis, tomato plants were sprayed a few times with a solution of this product during their growth. Results of the climate chamber test of the plants that were treated with Elot-Vis, showed a reduction of Botrytis cinerea infections on the inoculated leafs. Biological control agents seem to be not always very effective against Botrytis cinerea. The biological control agents that are containing micro organisms are also depending on the circumstances of the environment for an optimal development. These conditions are not always that optimal to compete with Botrytis cinerea or other micro organisms that are present on the plant.

Bacillus subtilis↗

Susceptibility of Vespula vulgaris (Hymenoptera: vespidae) to generalist entomopathogenic fungi and their potential for wasp control.

The pathogenicity of Vespula vulgaris wasp workers and larvae to a range of fungi was determined. All fungi were isolated in New Zealand and included isolates from Vespula, known generalist insect pathogens, and isolates generally nonpathogenic to insects. Workers and larvae were highly susceptible to pathogenic isolates at high spore concentrations (>1.75 x 10(5) cfu/individual). Eight isolates, two of Metarhizium anisopliae, five of Beauveria bassiana, and one of Aspergillus flavus were pathogenic while a single isolate of M. flavouiride var. novazealandicum, Cladosporium sp., and Paecilomyces sp. were not. The transfer of spores between workers, and between workers and larvae, was also investigated using several different application methods. Transfer of spores occurred between treated and untreated individuals, and for some of the application methods sufficient spores were transferred to cause mortality of the nontreated individuals. These findings are related to the potential of fungi for the control of wasps.

Animals↗

Characterization of an antifungal soil bacterium and its antagonistic activities against Fusarium species.

Bacteria were isolated from a cultivated soil and screened for antagonistic activity against Fusarium graminearum, a predominant agent of ear rot and head blight in cereal crops. Based on its in vitro effectiveness, isolate D1/2 was selected for characterization and identified as a strain of Bacillus subtilis by phenotypic tests and comparative analysis of its 16S ribosomal RNA gene (rDNA) sequence. It inhibited the mycelial growth of a collection of common fungal phytopathogens, including eight Fusarium species, three other ascomycetes, and one basidiomycete. The cell-free culture filtrate of D1/2 at different dilutions was active against macroconidium germination and hyphal growth of F. graminearum, depending on the initial macroconidium density. It induced the formation of swollen hyphal cells in liquid cultures of this fungus grown from macroconidia. A bioassay also demonstrated that D1/2 offered in planta protection against the damping-off disease in alfalfa seedlings caused by F. graminearum, while the type strain of B. subtilis was ineffective. Hence, B. subtilis D1/2 or its culture filtrate has potential application in controlling plant diseases caused by Fusarium.

Antibiosis↗

Larvicidal activity of blastospores and conidiospores of Beauveria bassiana (strain GK 2016) against age groups of Aedes aegypti.

Laboratory bioassays of two development stages, blastospores (BS) and conidiospores (CS) of Beauveria bassiana (strain GK 2016) against Aedes aegypti larvae were conducted at 27 degrees C. In Study 1, against 24 h post-hatched larvae, both BS and CS stages showed significant difference in their respective larvicidal efficacy over the control (P less than 0.0001). Larval mortality between 24 and 96 h post-exposure was significantly higher than any other time period investigated. Significantly higher larval mortality was observed with BS or CS at 10(8) ml-1 over lower concentrations (P less than 0.05). In Study 2, against different age groups, 12-24 h post-hatched larvae showed significantly higher mortality when treated with BS or CS than older age groups (P less than 0.05). A significant difference was found in the larvicidal potency of B. bassiana at different fungal stages.

Aedes↗

Effect of tadpole shrimp, Triops longicaudatus, (Notostraca: Triopsidae), on the efficacy of the microbial control agent Bacillus thuringiensis var. israelensis in experimental microcosms.

Laboratory bioassays using Culex quinquefasciatus larvae evaluated the effect of tadpole shrimp on the persistence of Bacillus thuringiensis var. israelensis (B.t.i.) in water collected from the surface of field microcosms. Time elapsed since B.t.i. treatment, as well as presence or absence of soil and tadpole shrimp, affected B.t.i. persistence at the water surface in 15- and 30-cm total depths. The presence of tadpole shrimp slowed the natural decline in B.t.i. effectiveness over time, but this effect was depressed when soil was present. Tadpole shrimp foraged throughout the water column and stirred up the substrate, keeping more particles in suspension at the surface than in microcosms with no shrimp, in microcosms with water depths of 15 and 30 cm.

Animals↗

Isolation of dipicolinic acid as an insecticidal toxin from Paecilomyces fumosoroseus.

Several entomopathogenic fungi produce toxins that could be used as bioinsecticides in integrated pest management programs. Paecilomyces fumosoroseus is currently used for the biological control of the whiteflies Bemisia tabaci and B. argentifolii. Supernatants from submerged batch culture, where the fungus produced abundant dispersed mycelium, conidia and blastospores, were toxic to the whitefly nymphs. The most abundant metabolite was purified by HPLC and identified by mass spectrometry and NMR as dipicolinic acid. Both the dipicolinic acid produced by the fungus and the chemically synthesized compound had insecticidal activity against third-instar nymphs of the insect. Dipicolinic acid was toxic to the whitefly nymphs in bioassays involving topical applications. In submerged culture, the specific growth rate of P. fumosoroseus was 0.054 h-1, the specific glucose consumption rate was 0.1195 g g-1 h-1 and the specific dipicolinic acid production rate was 0.00012 g g-1 h-1. Dipicolinic acid was detected after 24 h when the fungus started growing; and dipicolinic acid production was directly correlated with fungal growth. Nevertheless, the yield was low and the maximal concentration was only 0.041 g l-1. The maximal concentrations of conidia and blastospores (per milliliter) were 1.4x10(8) and 7x10(7), respectively.

Antibiosis↗

Two new bacterial pathogens of Colorado potato beetle Coleoptera: Chrysomelidae).

Other than Bacillus thuringiensis Berliner, few bacteria are lethal to the Colorado potato beetle (Leptinotarsa decemlineata [Say]), a major pest of potatoes and eggplant. Expanded use of biologicals for the control of Colorado potato beetle will improve resistance management, reduce pesticide use, and produce novel compounds for potential use in transgenic plants. Using freeze-dried, rehydrated artificial diet in pellet form to screen bacteria lethal to other insects, we determined that strains of Photorhabdus luminescens killed Colorado potato beetle larvae. The LC50 for second instar larvae of strain HM5-1 was 6.4 +/- 1.87 x 10(7) cells per diet pellet. In an attempt to find additional naturally occurring P. luminescens strains toxic to Colorado potato beetle larvae, we recovered, from soil, bacteria that produced a purple pigment. This bacterial strain, identified as Chromobacterium sp. by 16S ribosomal DNA sequencing, was also toxic to Colorado potato beetle larvae within 3 d. The LC50 for second instar larvae for these bacteria was 2.0 +/- 0.79 x 10(8) cells per diet pellet, while the LC50 was approximately 1 log lower for third instar larvae. P. luminescens appeared to kill by means of a protein toxin that may be similar to the described lepidopteran protein toxins. Based on the heat and acid stability, the toxin or toxins that Chromobacterium sp. produces, while not fully characterized, do not appear to be typical proteins. In both bacteria, the toxins are made after exponential growth ceases.

Animals↗

The Ecological Areawide Management (TEAM) of leafy spurge program of the United States Department of Agriculture-Agricultural Research Service.

The Ecological Areawide Management (TEAM) of Leafy Spurge program was developed to focus research and control efforts on a single weed, leafy spurge, and demonstrate the effectiveness of a coordinated, biologically based, integrated pest management program (IPM). This was accomplished through partnerships and teamwork that clearly demonstrated the advantages of the biologically based IPM approach. However, the success of regional weed control programs horizontally across several states and provinces also requires a vertical integration of several sectors of society. Awareness and education are the essential elements of vertical integration. Therefore, a substantial effort was made to produce a wide variety of information products specifically designed to educate different segments of society. During its tenure, land managers and agency decision makers have seen the potential of using the TEAM approach to accelerate the regional control of leafy spurge. The example set by the TEAM organization and participants is viewed as a model for future weed-control efforts.

Agriculture↗

Baseline susceptibility to bacterial insecticides in populations of Culex pipiens complex (Diptera: Culicidae) from California and from the Mediterranean Island of Cyprus.

Bacterial insecticides play an increasingly important role in mosquito control. To establish guidelines for detecting resistance at an early stage, information on natural variation in susceptibility of insect populations to these insecticides is needed. Between 1990 and 1993, the susceptibility of Culex pipiens L. complex to Bacillus thuringiensis subsp. israelensis de Barjac and/or Bacillus sphaericus Neide was determined in 31 collections from California. These collections were undertaken before the widespread use of B. thuringiensis subsp. israelensis and before the registration of B. sphaericus in California. Seven collections from the Mediterranean island of Cyprus, where no microbial insecticides have been used, also were tested. The 1990-1991 California collections exhibited limited variation in susceptibility to B. thuringiensis subsp. israelensis. LC50 and LC95 values spanned about a three-fold and four-fold range, respectively. The 1993 Cyprus collections exhibited both higher mean LC values, and greater variability in those values, than the California collections. The LC50s for the Cyprus collections varied over a 10-fold range, whereas the LC50s varied over a 12.5-fold range. Variation in susceptibility to B. sphaericus among the 1991 California collections was about five-fold at the LC50 and LC95. No significant geographic variation in susceptibility to B. thuringiensis subsp. israelensis was observed among regions within California. Although variation in susceptibility was limited among California collections, the greater variability observed among the Cyprus collections and between the Cyprus and California collections illustrates the importance of establishing regional baselines to monitor accurately for changes in susceptibility.

Animals↗

Different products for biological control of Botrytis cinerea examined on wounded stem tissue of tomato plants.

For the moment the agents that are used against Botrytis cinerea, in glasshouses were tomatoes are cultivated, are from chemical origin. For reducing the use of chemical agents in the future it is important to search for effective biological control agents against the fight of Botrytis cinerae. The following biological products Vital pasta, Vital gel and Elot-Vis were examined in there possibility to control Botrytis cinerea. Elot Vis was tested out in experiments that were carried out in climate chambers were leafs of 3 week old tomato plants were artificially infected with Botrytis cinerea spores. Also the biological products of Vital were first investigated in experiments that were carried out in climate chambers. In stead off leafs of tomato plants it were stem wounds of tomato plants who were treated with the pasta or the gel that was spread over the wounded surface after this has been inoculated with a suspension of conidia of Botrytis cinerae. The results of these first tests that were executed in the climate chambers were the circumstances for Botrytis cinerea were ideal seemed promising. In the next step these products were tested out on large scale in glasshouses. For each plant 5 wounds were created by removing the leafs, these wounds were or first treated with the Biological product and thereafter artificially infected with Botrytis cinerea spores to check out if these products can be used as a preventive agent or the wounds were first inoculated with a suspension of Botrytis cinerea spores and thereafter treated with the product. For the product Elot-Vis a few plants were totally sprayed with an Elot-Vis suspension before leafs were removed and the wounds were inoculated with conidia of Botrytis cinerea to check out if this product was able to activated the induced systemic resistance pathway. The experiments that were executed in glasshouses showed different percentages of succeeded Botrytis cinerea infections. This is probable due to the different weather conditions during both days that the experiments were executed. For the wounds that were treated with pasta it was difficult to distinguish wounds were Botrytis cinerea succeeded to infect the plant, because these wounds frequently didn't show any sign of infection on the surface but when the wounds with the pasta were cut open it was possible to see Botrytis cinerea infections inside the stem.

Biological Products↗

Laboratory and field tests of spray-dried and granular formulations of a Bacillus thuringiensis strain with insecticidal activity against the sugarcane borer.

Formulations of Bacillus thuringiensis Berliner (Bt) with insecticidal activity against the sugarcane borer, Diatraea saccharalis Fabricius (Lepidoptera: Pyralidae), were developed and tested under laboratory and field conditions. The formulations were prepared using biodegradable polymers such as modified corn starch as an encapsulating agent, gelatin as an adherent, powdered sugarcane as a feeding stimulant and a Bt var. kurstaki GM-34 strain from a non-sugarcane region as the active ingredient. The spore-crystal complex of this strain was mixed at three different concentrations (30, 70 and 100 g kg(-1)) with the other ingredients. The blends were prepared as spray-dried and granular formulations, and then submitted to laboratory tests with two day old larvae of D. saccharalis and field tests in sugarcane crops with natural sugarcane borer infestation. Spray-dried formulations in laboratory bioassays caused mortality near 100% with all three concentrations, and granular formulations caused mortality around 84%. The field tests showed that spray-dried formulations at 70 and 100 g kg(-1) concentrations were as effective as a commercial bioinsectide (Lepinox), while granular formulations were ineffective.

Animals↗