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Concept: Bacillus thuringiensis israelensis

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Mosquito control based on the use of Bacillus thuringiensis israelensis (Bti) is regarded as an environmental friendly method. However, Bti also affects non-target chironomid midges that are recognized as a central resource in wetland food webs. To evaluate the risk for different larval stages of Chironomus riparius we performed a test series of daily acute toxicity laboratory tests following OECD guideline 235 over the entire aquatic life cycle of 28 days. Our study is the first approach that performs an OECD approved test design with Bti and C. riparius as a standard organism in ecotoxicological testing. First-instar larvae of Chironomus riparius show an increased sensitivity towards Bti which is two orders of magnitude higher than for fourth instar larvae. Most EC50 values described in the literature are based on acute toxicity tests using third and fourth instar larvae. The risk for chironomids is underestimated when applying the criteria of the biocide regulation EU 528/2012 to our data and therefore the existing assessment approval is not protective. Possible impacts of Bti induced changes in chironomid abundances and community composition may additionally affect organisms at higher trophic levels, especially in spring when chironomid midges represent a key food source for reproducing vertebrates.

Concepts: Organism, Larva, Mosquito, Trophic level, Food chain, Bacillus thuringiensis, Chironomidae, Bacillus thuringiensis israelensis

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The cyanobacterium Anabaena PCC 7120#11 has been genetically engineered to act as a delivery vehicle for Bacillus thuringiensis subspecies israelensis mosquitocidal toxins. To address ecological concerns about releasing this genetically engineered microorganism into the environment for mosquito larva control, the persistence and ecological impacts of PCC 7120#11 was evaluated using multi-species, standardized aquatic microcosms.

Concepts: Bacteria, Biology, Microbiology, Ecology, Natural environment, Environmentalism, Bacillus thuringiensis, Bacillus thuringiensis israelensis

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The entomopathogenic Bacillus thuringiensis serovar israelensis displays peculiar conjugative transfer capabilities, accounted for by the large conjugative plasmid pXO16 (350 kb). The efficient and fast conjugative transfers are accompanied by a macroscopic aggregation of bacterial partners. Moreover, pXO16 has proven capable of effective mobilization and the retro-transfer of both mobilizable and “non-mobilizable” plasmids. In this work, the aggregation phenomenon is shown to promote pXO16 transfer while not being mandatory for transfer. Transfer of pXO16 to B. thuringiensis recipient strains that do not display aggregation is observed as well, hence enlarging the previously defined host range. The use of variant calling analysis of transconjugants allowed for observation of up to 791 kb chromosomal regions mobilization. Previous analysis of pXO16 did not reveal any type IV secretion system (T4SS) homologs, which suggested the presence of an unusual conjugative system. A FtsK/SpOIIIE ATPase gene proved here to be necessary for conjugative transfer. Additionally, the analysis of natural restriction-modification systems in both conjugative partners gave credit to a ssDNA transfer mechanism. A “transfer israelensis plasmid” (tip) region containing this ATPase gene was shown to code for other potential T4SS proteins, illustrating a conjugative system distantly related to the other known Gram-positive T4SSs. This article is protected by copyright. All rights reserved.

Concepts: DNA, Gene, Cell, Bacteria, Microbiology, Secretion, Plasmid, Bacillus thuringiensis israelensis

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The annual incidence of dengue has been increasing over the last few years in Sri Lanka with seasonal epidemics. Biological control of the vector has not been part of the integrated vector control implemented by the public health authorities of Sri Lanka so far. This pilot study assessed the effectiveness of using Bacillus thuringiensis israelensis (Bti) spray to control the Aedes mosquito vector population density.

Concepts: Epidemiology, Mosquito, Aedes, Dengue fever, Sri Lanka, Mosquito control, Bacillus thuringiensis, Bacillus thuringiensis israelensis

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Due in part to the development of insecticide resistance, the common bed bug, Cimex lectularius, has overcome human intervention efforts to make a global resurgence. The failure of chemical pesticides has created a need for novel strategies to combat bed bugs. While a number of insect pests are susceptible to the use of entomopathogenic microbes or microbial-derived toxins, biological control methods have not been thoroughly explored in bed bugs. Here, we tested the virulence of three entomopathogenic bacterial species in C. lectularius to determine their potential for bed bug control. We examined bed bug survival after inoculation with live or heat-killed Serratia marcescens, Pseudomonas fluorescens, and Bacillus thuringiensis israelensis at varying temperatures. We also analyzed the viability and growth of the same bacteria in infected bed bugs. All three bacterial species were pathogenic to bed bugs. However, the effects of S. marcescens and P. fluorescens were temperature-dependent while the lethality of B. thuringiensis israelensis was not. In addition, bacterial virulence was partly dependent on the route of infection but was not strongly associated with proliferation. Thus, our results suggest multiple possible mechanisms of microbial pathogenicity in the bed bug and indicate that entomopathogenic bacteria, or products derived from them, may have useful applications for bed bug control.

Concepts: Bacteria, Microbiology, Pesticide, Bacillus thuringiensis, Biological pest control, Bedbug, Cimex lectularius, Bacillus thuringiensis israelensis

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Pesticides are regularly used for a variety of applications and are disseminated throughout the environment. These substances may have significant negative impacts. To date, the half-life, t1/2, was often used to study the fate of pesticides in environmental matrices (water, soil, sediment). However, this value gives limited information. First, it does not evaluate the formation of by-products, resulting in the need for additional experiments to be performed to evaluate biodegradation and biotransformation products. T1/2 also fails to consider the chemical’s impact on biodiversity. Resilience time, a new and integrative proxy, was recently proposed as an alternative to t1/2, with the potential to evaluate all the post-application effects of the chemical on the environment. The ‘Environmental Metabolic Footprinting’ (EMF) approach, giving an idea of the resilience time, was used to evaluate the impact of botanicals on soil. The goal is to optimise the EMF to study the impact of a microbial insecticide, the Bacillus thuringiensis israelensis (Bti), on sediment. The difficulty of this work lies in the commercial solution of Bti that is really complex, and this complexity yields chromatograms that are extremely difficult to interpret; t1/2 cannot be used. No methodologies currently exist to monitor the impact of these compounds on the environment. We will test the EMF to determine if it is sensitive enough to tolerate such complex mixtures. A pure chemical insecticide, the α-cypermethrin, will be also studied. The article shows that the EMF is able to distinguish meta-metabolome differences between control and exposed (with Bti) sediments.

Concepts: Bacteria, Environment, Natural environment, Environmentalism, Environmental science, Bacillus thuringiensis, Insecticide, Bacillus thuringiensis israelensis

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pXO16, a large plasmid originating from Bacillus thuringiensis serovar israelensis, displays unique conjugation capacities: besides efficient self-transfer, it is able to mobilize and retro-mobilize non-conjugative plasmids, including those missing an oriT and/or a mob gene, also known as “non-mobilizable” plasmids. In this paper, another peculiar transfer property of pXO16 is described. This element is indeed able to transfer chromosomal loci at frequencies of ca. 10(-5)-10(-6) transconjugants/donor cell. Whereas most other chromosomal transfer systems occur via the integration of the conjugative elements into the chromosome prior to its transfer, pXO16 appears to transfer the chromosomal markers in the absence of physical integration, but rather through a “donation-type” mobilization.

Concepts: DNA, Gene, Cell, Bacteria, Chromosome, Plasmid, Triparental mating, Bacillus thuringiensis israelensis

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A multi-phased study was conducted in Cambodia from 2005-2011 to measure the impact of larviciding with the bacterial larvicide, Bacillus thuringiensis israelensis (Bti), a water dispersible granule (WG) formulation on the vector, Aedes aegypti (L.) and the epidemiology. In our studies, all in-use containers were treated at 8 g/1000 L, including smaller containers and animal feeders which were found to contribute 23% of Ae aegypti pupae. The treated waters were subjected to routine water exchange activities. Pupal production was suppressed by an average 91% for 8 weeks. Pupal numbers continued to remain significantly lower than the untreated commune (UTC) for 13 weeks post treatment in the peak dengue vector season (p<0.05). Suppression of pupal production was supported by very low adult numbers in the treated commune. An average 70% of the household harbored 0-5 Ae aegypti mosquitoes per home for 8 weeks post treatment, but in the same period of time >50% of the household in the UTC harbored ≥11 mosquitoes per home. The adult population continued to remain at significantly much lower numbers in the Bti treated commune than in the UTC for 10-12 weeks post treatment (p<0.05). In 2011, a pilot operational program was evaluated in Kandal Province, a temephos resistant site. It was concluded that 2 cycles of Bti treatment in the 6 months monsoon season with complete coverage of the target districts achieved an overall dengue case reduction of 48% in the 6 treated districts compared to the previous year, 2010. Five untreated districts in the same province had an overwhelming increase of 352% of dengue cases during the same period of time. The larvicide efficacy, treatment of all in-use containers at the start of the monsoon season, together with treatment coverage of entire districts interrupted disease transmission in the temephos resistant province.

Concepts: Time, Mosquito, Yellow fever, Aedes aegypti, Aedes, Dengue fever, Bacillus thuringiensis, Bacillus thuringiensis israelensis

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Bacillus thuringiensis δ-endotoxins are the most widely used biopesticides for controlling economically important crop pests and disease vectors. Improving its efficacy is of great benefit. Here, an improvement of Cry2Aa δ-endotoxins toxicity was conducted via a cry gene over-expression system using P20 from B. thuringiensis israelensis.

Concepts: Caterpillar, Bacillus thuringiensis, Bacillaceae, Bacillus thuringiensis israelensis

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Bacillus thuringiensis serovar finitimus strain YBT-020 is the well-studied spore-crystal association (SCA) phenotypic strain, whose parasporal crystals adhere to spore after lysis of the mother cell. Its endogenous plasmids pBMB26 and pBMB28 were proved essential for this SCA phenotype. In our previous study, using conventional methods, pBMB26 cured derivative and both pBMB26 and pBMB28 cured derivative of YBT-020 were obtained. However, YBT-020 solely cured of pBMB28 could not be obtained. In this study, an unstable replication region of pBMB28 was identified and was used to construct an incompatible plasmid pRep28B. This incompatible plasmid was successfully used to cure plasmid pBMB28 and was easily eliminated through segregational instability under the optimum growth temperature of YBT-020. Therefore, an endogenous plasmid was cured from the B. thuringiensis strain utilizing plasmid incompatibility. Moreover, using an unstable replication region instead of a temperature sensitive (Ts) replication region is better to cure the incompatible plasmid because it can avoid culturing at higher temperature. This method provides an efficient method for plasmid curing in B. thuringiensis and other bacteria.

Concepts: Gene, Cell, Bacteria, Microbiology, Plasmid, Bacillus, Bacillus thuringiensis, Bacillus thuringiensis israelensis