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Bacillus thuringiensis (Bt) Berliner has potential for use in insect management. Its use can be an alternative for the management of Bradysia aff. ocellaris (Comstock), considered one of the main strawberry pests in a soilless system. Therefore, the objective of this work was to evaluate the toxicity of different bacteria on B. aff. ocellaris in laboratory and greenhouse bioassays. The following isolates were used in the experiments: Bacillus circulans (Bc), B. thuringiensis var. oswaldo cruzi (Bto) or B. thuringiensis var. israelensis (Bti) and B. thuringiensis var. kurstaki (Btk) In the laboratory, B. aff. ocellaris larvae showed high susceptibility to Bti isolate (92 % mortality) 14 days after treatment exposure (DAET). In contrast, the isolates Bc, Bto, and Btk showed less than 32 % mortality, not differing from the control treatment (water ­ 22 % mortality). According to the concentration-response curves the values of lethal concentration LC50 and LC90 were 4 x 106 CFU.mL-1 and 4 x 1015 CFU.mL-1. By applying Bti (4 × 1012 CFU.mL-1) at the base of strawberry plants growing in plastic pots containing commercial plant substrate, a reduction of 26 % in the emergence of B. aff. ocellaris was observed. According to these results, the Bti isolate is considered promising for the formulation of bioinsecticides based on Bt for the management of B. aff. ocellaris in strawberry culture.

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Bacillus thuringiensis svar. israelensis (Bti) larvicides are effective in controlling Aedes aegypti; however, the effects of long-term exposure need to be properly evaluated. We established an Ae. aegypti strain that has been treated with Bti for 30 generations (RecBti) and is still susceptible to Bti, but females exhibited increased susceptibility to Zika virus (ZIKV). This study compared the RecBti strain to a reference strain regarding: first, the relative transcription of selected immune genes in ZIKV-challenged females (F30) with increased susceptibility detected in a previous study; then, the whole transcriptomic profile using unchallenged females (F35). Among the genes compared by RT-qPCR in the ZIKV-infected and uninfected females from RecBti (F30) and the reference strain, hop, domeless, relish 1, defensin A, cecropin D, and gambicin showed a trend of repression in RecBti infected females. The transcriptome of RecBti (F35) unchallenged females, compared with a reference strain by RNA-seq, showed a similar profile and only 59 differentially expressed genes were found among 9202 genes analyzed. Our dataset showed that the long-term Bti exposure of the RecBti strain was associated with an alteration of the expression of genes potentially involved in the response to ZIKV infection in challenged females, which is an important feature found under this condition.

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Larvicides based on the bacteria Bacillus thuringiensis svar. israelensis (Bti) and Lysinibacillus sphaericus are effective and environmentally safe compounds for the control of dipteran insects of medical importance. They produce crystals that display specific and potent insecticidal activity against larvae. Bti crystals are composed of multiple protoxins: three from the three-domain Cry type family, which bind to different cell receptors in the midgut, and one cytolytic (Cyt1Aa) protoxin that can insert itself into the cell membrane and act as surrogate receptor of the Cry toxins. Together, those toxins display a complex mode of action that shows a low risk of resistance selection. L. sphaericus crystals contain one major binary toxin that display an outstanding persistence in field conditions, which is superior to Bti. However, the action of the Bin toxin based on its interaction with a single receptor is vulnerable for resistance selection in insects. In this review we present the most recent data on the mode of action and synergism of these toxins, resistance issues, and examples of their use worldwide. Data reported in recent years improved our understanding of the mechanism of action of these toxins, showed that their combined use can enhance their activity and counteract resistance, and reinforced their relevance for mosquito control programs in the future years.

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Insecticide resistance has become problematic in tropical and subtropical regions, where Aedes mosquitoes and Aedes-borne arboviral diseases thrive. With the recent occurrence of chikungunya and the Zika virus in Jamaica, the Ministry of Health and Wellness, Jamaica, partnered with the United States Agency for International Development to implement multiple intervention activities to reduce the Aedes aegypti populations in seven parishes across the island and to assess the susceptibility of collected samples to various concentrations of temephos, Bacillus thuringiensis subsp. israelensis, (Bti), diflubenzuron and methoprene. Of the insecticides tested, only temephos has been used in routine larviciding activities in the island. The results showed that only temephos at concentrations 0.625 ppm and Bti at concentrations 6-8 ppm were effective at causing 98-100% mortality of local Ae. aegypti at 24 h exposure. Surprisingly, the growth inhibitors diflubenzuron and methoprene had minimal effect at preventing adult emergence in Ae. aegypti larvae in the populations tested. The results demonstrate the need for insecticide resistance testing as a routine part of vector control monitoring activies in order to determine useful tools that may be incorporated to reduce the abundance of Ae. aegypti.

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The chemical control of the mosquito Aedes aegypti is a great challenge worldwide, since several populations of this species are already resistant to traditional insecticides, such as temephos. In Brazil, alternative larvicides, such as Bacillus thuringiensis israelensis (Bti) and pyriproxyfen, have been used more recently. In this study we evaluated the persistence of pyriproxyfen (Sumilarv 0.5%G), 2 commercial formulations of Bti (Vectobac WDG and Vectobac G), and temephos (Fersol 1G) under field and simulated field conditions with treatments exposed to sun and shadow. In the field tests, the 2 formulations of Bti presented less persistence in the 8th wk of evaluation (46% and 37% positivity) compared with temephos (3.6% and 6.8% positivity) and Sumilarv (6.6% and 3.8% positivity) in containers exposed to the sun and shadow, respectively. In the simulated field trial, temephos and the 2 formulations of Bti presented high persistence (100% mortality at 8th wk) when applied in the water box and in deposits placed in the shade. In containers exposed to the sun, the persistence of these products was lower (>80% mortality by temephos after 4 wk, Bti formulations for 3 wk, and Sumilarv for 6 wk). Based on these data, however, Sumilarv presented better performance in the containers exposed to the sun than the 2 formulations of Bti (Vectobac G and Vectobac WDG), which were affected by the sun.

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OBJECTIVE: To evaluate combinations of larvicides and fish which are predators of larvae of Aedes aegypti, namely Betta splendens, Trichogaster trichopterus and Poecilia reticulata. METHODS: We used 15 water tanks with a capacity of 250 litres of water. In 10 tanks, larvicide - Temephos, Bti and Novaluron - was added, the other five contained only one specimen of fish and unchlorinated water. The fish were monitored for a week without changing the water, and their survival recorded on a form. An estimate of the Kaplan-Meier survival was performed to determine the significance of the tests. RESULTS: Betta splendens showed the lowest mortality range of larvicides tested. CONCLUSION: Combined use of larvivorous fish and larvicides in large water tanks is feasible.

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Aedes aegypti has played a major role in the dramatic expansion of dengue worldwide. The failure of control programs in reducing the rhythm of global dengue expansion through vector control suggests the need for studies to support more appropriated control strategies. We report here the results of a longitudinal study on Ae. aegypti population dynamics through continuous egg sampling aiming to characterize the infestation of urban areas of a Brazilian oceanic island, Fernando de Noronha. The spatial and temporal distribution of the dengue vector population in urban areas of the island was described using a monitoring system (SMCP-Aedes) based on a 103-trap network for Aedes egg sampling, using GIS and spatial statistics analysis tools. Mean egg densities were estimated over a 29-month period starting in 2011 and producing monthly maps of mosquito abundance. The system detected continuous Ae. aegypti oviposition in most traps. The high global positive ovitrap index (POI=83.7% of 2815 events) indicated the frequent presence of blood-fed-egg laying females at every sampling station. Egg density (eggs/ovitrap/month) reached peak values of 297.3 (0 - 2020) in May and 295 (0 - 2140) in August 2012. The presence of a stable Ae. aegypti population established throughout the inhabited areas of the island was demonstrated. A strong association between egg abundance and rainfall with a 2-month lag was observed, which combined with a first-order autocorrelation observed in the series of egg counts can provide an important forecasting tool. This first description of the characteristics of the island infestation by the dengue vector provides baseline information to analyze relationships between the spatial distribution of the vector and dengue cases, and to the development of integrated vector control strategies.

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The aim of this work was to study the production of the bioinsecticide using the aerobic cultivation. Two-stage cultivation process was used: aerobic vegetative growth and end-phase of sporulation (with aeration and without aeration). The cuts of working volume evaluated were 20, 40, 60 and 80 percent. The highest value of spore concentration (2.17x10(9) CFU/mL) and spore productivity (7.2x10(7) CFU/mL.h) were observed using SP60. The lethal concentration (CL50) obtained through the aeration process and non-aeration process were 0.21 and 0.05 mg/L, respectively. The specific potential of the bioinsecticide produced without aeration (0.025 mg/10-12 CFU) was two times higher than that obtained with aeration (0.05 mg/10-12 CFU).

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Aedes aegypti is the vector of Dengue disease, responsible for 20,000 deaths/year worldwide. Bacillus thuringiensis var israelensis - Bti releases selective and effective toxins (crystal proteins) against A. aegypti larvae. We present a low cost bioprocess for toxin production, accomplished by a selected Brazilian strain Bti (BR-LPB01) and employment of low cost substrates. Soybean meal and sugarcane molasses lead to high toxic effectiveness after 2L bioreactor fermentation (LD50=26ng/mL), near to the reference strain IPS82 (LD50=17.3 ng/mL). The pH ranged between 5.8 and 7.0 during the exponential growth period and between 7.0 and 8.4 during the stationary phase, with low activity. Thus, control of foam and pH 7.0 were started and proved to be crucial for high activity. It was verified that the fermentation could be discontinued after 20 hours, when the highest activity was present.

A dengue é transmitida pelo Aedes aegypti, doença responsável por 20.000 mortes/ano no mundo. Bacillus thuringiensis var israelensis libera toxinas seletivas e eficazes (proteínas cristal) contra larvas de A. aegypti. Propõe-se um bioprocesso de baixo custo para a produção da toxina, pelo emprego de uma cepa brasileira selecionada de Bti (BR-LPB01) e de substratos de baixo custo. Farelo de soja e melaço de cana levaram a eficácia tóxica alta após fermentação em biorreator 2L (DL50=26ng/mL), valor próximo a estirpe de referência IPS82 (DL50=17,3 ng/mL). O pH variou entre 5,8 e 7,0 durante o período de crescimento exponencial e entre 7,0 e 8,4 durante a fase estacionária, com baixa atividade larvicida. Assim, controles de espuma e de pH 7,0 foram iniciados e demonstraram serem cruciais para alta atividade. Verificou-se que a fermentação deve ser interrompida após vinte horas, quando se obtém a maior atividade.

RESUMO

A dengue é atualmente considerada a mais importante arbovirose que afeta o homem. O agravamento desta situação epidemiológica tem acarretado um aumento expressivo no uso de inseticidas organofosforados no combate ao vetor, principalmente o temefós, que é amplamente e sistematicamente aplicado em ambientes urbanos em reservatórios de água, até para o consumo humano. Este projeto propôs investigar efeito genotóxico (clastogênico) em células de mamíferos, induzido pela exposição ao temefós (grau técnico 95,5%), e ao biolarvicida Bacillus thuringiensis sorovar israelenses (Bti) - IPS 82, ambos empregados no controle do Aedes aegypti. Na avaliação genotóxica foram utilizadas células da medula óssea de camundongos albinos Swiss Webster de ambos os sexos, empregando-se os testes de metáfase e de micronúcleos para detecção dos danos cromossômicos: aberrações cromossômicas e micronúcleos. Os camundongos foram tratados com temefós (grau técnico 95,5%) nas concentrações de 27,75; 55,5 e 111 mg/kg ou com água destilada 10 ml/kg, via gavagem, como controle negativo ou com Ciclofosfamida a 25 mg/kg, via i.p., como controle positivo por 24, 48 e 72h, em dose única ou em 9 doses repetidas (1 dose/semana). Outros grupos foram tratados com Bti nas doses de 204 e 122,4 UFC (Unidade Formadora de Colônia) ou água destilada (200 ?l) como controle negativo, via gavagem, por 24 e 48h. Os resultados observados confirmaram a ação genotóxica induzida pelo inseticida temefós em camundongos de ambos os sexos, com a formação de micronúcleo em eritrócitos policromáticos (PCE MN), em todas as concentrações testadas 24h após o tratamento único. Na concentração de 111,00 mg/kg induziu PCE MN também após 48 e 72h em dose única e após tratamento com 9 doses. O temefós induziu também aberrações cromossômicas nas células em metáfases, em todas as concentrações testadas, 24h após tratamento único, e na concentração de 111,00 mg/kg, também após 48 e 72h de tratamento...

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A new approach to dengue vector surveillance based on permanent egg-collection using a modified ovitrap and Bacillus thuringiensis israelensis(Bti) was evaluated in different urban landscapes in Recife, Northeast Brazil. From April 2004 to April 2005, 13 egg-collection cycles of four weeks were carried out. Geo-referenced ovitraps containing grass infusion, Bti and three paddles were placed at fixed sampling stations distributed over five selected sites. Continuous egg-collections yielded more than four million eggs laid into 464 sentinel-ovitraps over one year. The overall positive ovitrap index was 98.5 percent (over 5,616 trap observations). The egg density index ranged from 100 to 2,500 eggs per trap-cycle, indicating a wide spread and high density of Aedes aegypti (Diptera: Culicidae) breeding populations in all sites. Fluctuations in population density over time were observed, particularly a marked increase from January on, or later, according to site. Massive egg-collection carried out at one of the sites prevented such a population outbreak. At intra-site level, egg counts made it possible to identify spots where the vector population is consistently concentrated over the time, pinpointing areas that should be considered high priority for control activities. The results indicate that these could be promising strategies for detecting and preventing Ae. aegypti population outbreaks.

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