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1.
Braz. j. biol ; 84: e256933, 2024. tab, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1364505

RESUMO

Anticarsia gemmatalis Hünber, 1818 is one of the main defoliating species in the soybean crop. Bacillus thuringiensis Berliner, 1915, is a bacterium used in the biological control of this pest species. Resistant populations and their sublethal effects caused by the use of the bacteria have already been reported; however, there are no studies on phenotypic plasticity in adulthood exposed to Bt-based bioinsecticide sub-doses. This study aimed to evaluate the morphometry of A. gemmatalis adults under laboratory conditions submitted to the Bt-based bioinsecticide Dipel® over the three generations. The body segments mensuread were width, length, and area of the anterior and posterior wings, the weight of the integument, chest, abdomen, wings, and the whole adult of males and females. Among the treatments, LC5 in the first generation and LC10 in the second generation were those with lower thresholds in relation to the weight of the chest and abdomen, considering the proportions of the body smaller than the females. The female's weight adulthood was reduced by 10% about males, and, only in the first generation. Males have larger body size and more pronounced phenotypic plasticity than females. Here, we demonstrate the first study assessing the phenotypic plasticity of A. gemmatalis adults.


Anticarsia gemmatalis Hünber, 1818 é uma das principais espécies desfolhadoras da cultura da soja. Bacillus thuringiensis Berliner, 1915, é uma bactéria utilizada no controle biológico dessa espécie de praga. Populações resistentes e seus efeitos subletais causados pelo uso da bactéria já foram relatados, no entanto, não há estudos sobre a plasticidade fenotípica na idade adulta exposta a subdoses de bioinseticida à base de Bt. Este trabalho teve como objetivo avaliar a morfometria de adultos de A. gemmatalis em condições de laboratório submetidos ao bioinseticida Dipel® ao longo de três gerações. Os segmentos corporais mensuráveis eram largura, comprimento e área das asas anterior e posterior, o peso do tegumento, tórax, abdômen, asas e todo o adulto de machos e fêmeas. Dentre os tratamentos, CL5 na primeira geração e CL10 na segunda geração foram aqueles com limiares mais baixos em relação ao peso do tórax e abdômen, considerando as proporções do corpo menores que as do sexo feminino. O peso da fêmea na idade adulta foi reduzido em 10% em relação aos machos e, apenas na primeira geração. Os machos têm tamanho corporal maior e plasticidade fenotípica mais pronunciada do que as fêmeas. Este estudo demonstra o primeiro estudo avaliando a plasticidade fenotípica de adultos de A. gemmatalis.


Assuntos
Animais , Fenótipo , Soja , Bacillus thuringiensis , Controle Biológico de Vetores
2.
Environ Monit Assess ; 195(2): 257, 2023 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-36595073

RESUMO

Analytical methods exist to detect biothreat agents in environmental samples during a response to biological contamination incidents. However, the coastal zone facilities and assets of the US Coast Guard (USCG), including response boats in diverse geographical areas and maritime environmental conditions, can pose complex and unique challenges for adapting existing analytical detection methods. The traditional culture (TC) and the rapid viability polymerase chain reaction (RV-PCR) methods were evaluated for their compatibility for maritime environmental surface and grab sample analysis to detect spores of Bacillus thuringiensis subspecies kurstaki (Btk), a surrogate for Bacillus anthracis. The representative samples collected from a USCG installation included surfaces, such as aluminum on boats, nonskid tread on decks of watercraft, computer touchscreens, and concrete piers, and grab samples of boat washdown water, soil, vegetation, and gravel from surrounding areas. Replicate samples were spiked with Btk spores at two to three tenfold increasing levels and analyzed. Out of a total of 150 samples collected and analyzed, the TC method gave 10 false-positive and 19 false-negative results, while the RV-PCR method-based analysis resulted in 0 false-positive and 26 false-negative results. An abundance of microbial background and particulates in some samples interfered with true results, while both methods gave similar results for samples with low microbial background and particulates. Improved and high-throughput sample processing methods are needed for analysis of complex environmental samples.


Assuntos
Bacillus anthracis , Bacillus thuringiensis , Bacillus anthracis/genética , Esporos Bacterianos , Monitoramento Ambiental , Reação em Cadeia da Polimerase/métodos
3.
Ecotoxicol Environ Saf ; 250: 114503, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-36610297

RESUMO

Emerging aquatic insects serve as one link between aquatic and adjacent riparian food webs via the flux of energy and nutrients. These insects provide high-quality subsidy to terrestrial predators. Thus, any disturbance of emergence processes may cascade to higher trophic levels and lead to effects across ecosystem boundaries. One stressor with potential impact on non-target aquatic insects, especially on non-biting midges (Diptera: Chironomidae), is the widely used mosquito control agent Bacillus thuringiensis var. israelensis (Bti). In a field experiment, we investigated emerging insect communities from Bti-treated (three applications, maximum field rate) and control floodplain pond mesocosms (FPMs) over 3.5 months for changes in their composition, diversity as well as the emergence dynamics and the individual weight of emerged aquatic insects over time. Bti treatments altered community compositions over the entire study duration - an effect mainly attributed to an earlier (∼10 days) and reduced (∼26%) peak in the emergence of Chironomidae, the dominant family (88% of collected individuals). The most reasonable explanation for this significant alteration is less resource competition caused by a decrease in chironomid larval density due to lethal effects of Bti. This is supported by the higher individual weight of Chironomidae emerging from treated FPMs (∼21%) during Bti application (April - May). A temporal shift in the emergence dynamics can cause changes in the availability of prey in linked terrestrial ecosystems. Consequently, terrestrial predators may be affected by a lack of appropriate prey leading to bottom-up and top-down effects in terrestrial food webs. This study indicates the importance of a responsible and elaborated use of Bti and additionally, highlights the need to include a temporal perspective in evaluations of stressors in aquatic-terrestrial meta-ecosystems.


Assuntos
Bacillus thuringiensis , Chironomidae , Humanos , Animais , Ecossistema , Cadeia Alimentar , Insetos
4.
Arch Microbiol ; 205(1): 53, 2023 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-36602580

RESUMO

The bacterial communities that colonize the cadaver environment of insects infected and killed by parasitic fungi can be selected by the sympatric fungi and provide novel impacts. In this study, we found that Bacillus cereus/thuringiensis predominate the bacterial community in Dolichoderus thoracicus ant cadavers colonized by O. pseudolloydii. The most predominant bacterial strains in these ant cadavers were hemolytic and able to produce hydrolytic enzymes for digesting the ant tissue. A relatively intense lethal effect on the co-cultured nematode was displayed by a hemolytic strain. Moreover, the antagonistic effect against pathogenic fungi detected in the bacteria sympatric with O. pseudolloydii was reported here. Naphthoquinones have been shown to confer antibacterial activities and produced by the ant-pathogenic Ophiocordyceps fungi. However, our results did not show the naphthoquinone tolerance we expected to be detected in the bacteria from the ant infected by O. pseudolloydii. The bacterial diversity in the samples associated with O. pseudolloydii infected ants as revealed in this study will be a step forward to the understanding of the roles playing by the microbial community in the native habitats of O. pseudolloydii.


Assuntos
Formigas , Bacillus thuringiensis , Bacillus , Animais , Formigas/microbiologia , Insetos
5.
PeerJ ; 11: e14670, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36684673

RESUMO

Bacillus thuringiensis, known to be one of the most important biocontrol microorganisms, contains three AA10 family lytic polysaccharide monooxygenases (LPMOs) in its genome. In previous reports, two of them, BtLPMO10A and BtLPMO10B, have been preliminarily characterized. However, some important biochemical features and substrate preference, as well as their potential applications in chitin degradation, still deserve further investigation. Results from present study showed that both BtLPMO10A and BtLPMO10B exhibit similar catalytic domains as well as highly conserved substrate-binding planes. However, unlike BtLPMO10A, which has comparable binding ability to both crystalline and amorphous form of chitins, BtLPMO10B exhibited much stronger binding ability to colloidal chitin, which mainly attribute to its carbohydrate-binding module-5 (CBM5). Interestingly, the relative high binding ability of BtLPMO10B to colloidal chitin does not lead to high catalytic activity of the enzyme. In contrast, the enzyme exhibited higher activity on ß-chitin. Further experiments showed that the binding of BtLPMO10B to colloidal chitin was mainly non-productive, indicating a complicated role for CBM5 in LPMO activity. Furthermore, synergistic experiments demonstrated that both LPMOs boosted the activity of the chitinase, and the higher efficiency of BtLPMO10A can be overridden by BtLPMO10B.


Assuntos
Bacillus thuringiensis , Oxigenases de Função Mista , Oxigenases de Função Mista/genética , Bacillus thuringiensis/genética , Polissacarídeos/metabolismo , Quitina/química , Domínio Catalítico
6.
Sci Rep ; 13(1): 831, 2023 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-36646757

RESUMO

Monitoring the presence of pathogenic Bacillus spores is important for industrial applications, as well as necessary for ensuring human health. Bacillus thuringiensis is used as a biopesticide against several insect pests. Bacillus cereus spores are a significant cause of food poisoning, and Bacillus anthracis is a recognized biosecurity threat. Laboratory-based methods, such as polymerase chain reaction, enzyme-linked immunosorbent assay, or matrix-assisted laser desorption ionization spectroscopy provide sensitive detection of bacteria and spores, but the application of those methods for quasi-continuous environmental monitoring presents a significant challenge requiring frequent human intervention. To address this challenge, we developed a workstation for quasi-autonomous monitoring of water reservoirs for the presence of bacteria and spores, and designed and validated the functionality of a microprocessor-controlled module capable of repetitive collection and pre-concentration of spores in liquid samples tested with fiberglass (FG), polyether sulfone and polyvinylidene fluoride filters. The best results were obtained with FG filters delivering a 20× concentration of B. thuringiensis and B. cereus spores from saline suspensions. The successful 20× pre-concentration of Bacillus spores demonstrated with FG filters could be repeated up to 3 times when bleach decontamination is applied between filtrations. Taken together, our results demonstrate an attractive instrument suitable for semi-automated, quasi-continuous sampling and pre-processing of water samples for biosensing of bacterial spores originating from a complex environment.


Assuntos
Bacillus anthracis , Bacillus thuringiensis , Bacillus , Humanos , Bacillus cereus , Esporos Bacterianos/química
7.
Toxins (Basel) ; 15(1)2023 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-36668875

RESUMO

The beetle Anthonomus grandis Boheman, 1843, is the main cotton pest, causing enormous losses in cotton. The breeding of genetically modified plants with A. grandis resistance is seen as an important control strategy. However, the identification of molecules with high toxicity to this insect remains a challenge. The susceptibility of A. grandis larvae to proteins (Cry1Ba, Cry7Ab, and Mpp23Aa/Xpp37Aa) from Bacillus thuringiensis Berliner, 1915, with toxicity reported against Coleopteran, has been evaluated. The ingestion of different protein concentrations (which were incorporated into an artificial diet) by the larvae was tested in the laboratory, and mortality was evaluated after one week. All Cry proteins tested exhibited higher toxicity than that the untreated artificial diet. These Cry proteins showed similar results to the control Cry1Ac, with low toxicity to A. grandis, since it killed less than 50% of larvae, even at the highest concentration applied (100 µg·g-1). Mpp/Xpp proteins provided the highest toxicity with a 0.18 µg·g-1 value for the 50% lethal concentration. Importantly, this parameter is the lowest ever reported for this insect species tested with B. thuringiensis proteins. This result highlights the potential of Mpp23Aa/Xpp37Aa for the development of a biotechnological tool aiming at the field control of A. grandis.


Assuntos
Bacillaceae , Bacillales , Bacillus thuringiensis , Besouros , Inseticidas , Gorgulhos , Animais , Larva , Bacillus thuringiensis/genética , Bacillus thuringiensis/metabolismo , Inseticidas/toxicidade , Inseticidas/metabolismo , Melhoramento Vegetal , Gossypium
8.
Pestic Biochem Physiol ; 188: 105269, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36464374

RESUMO

Insect resistance to Bacillus thuringiensis (Bt) is a critical limiting factor for applying the Bt crops. Some studies indicated that decreased protoxin activation because of lower enzymatic activities of trypsin and chymotrypsin and increased expression of serpin might involve in Bt resistance. Our previous study identified an endogenous serpin could inhibit the midgut proteases to activate Cry1Ac and reduce the insecticide activity to Helicoverpa armigera. We hypothesis that up-regulated serpin involve in resistance via inhibiting enzymatic activities of trypsin and chymotrypsin to decrease protoxin activation. Herein, we found the serpin-e gene relative expression in midgut was significantly higher in the LF30 resistant strain than that in the susceptible strain during all developmental stages. Importantly, RNAi-mediated silencing of serpin-e gene expression caused 4.46-fold mortality changes in LF30 strain, but the trypsin and chymotrypsin proteases activities were only changed 0.79-fold and 2.22-fold. In addition, although proteases activities were significantly lower in LF30 strain than that in the susceptible strain, the resistance ratios of LF30 to Cry1Ac protoxin and to activated Cry1Ac toxin were no difference. The results indicated serpins caused insect resistance to Cry1Ac protoxins partly through inhibiting the trypsin and chymotrypsin proteases activities, but it also existed other mechanisms in LF30.


Assuntos
Bacillus thuringiensis , Mariposas , Serpinas , Animais , Serpinas/genética , Quimotripsina/genética , Tripsina , Peptídeo Hidrolases , Mariposas/genética
9.
Front Immunol ; 13: 1022445, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36466886

RESUMO

Midgut receptors have been recognized as the major mechanism of resistance to Cry proteins in lepidopteran larvae, while there is a dearth of data on the role of hemocyte's response to Cry intoxication and resistance development. We aimed at investigating the role of circulating hemocytes in the intoxication of Cry1F toxin in larvae from susceptible (ACB-BtS) and resistant (ACB-FR) strains of the Asian corn borer (ACB), Ostrinia furnacalis. Transcriptome and proteome profiling identified genes and proteins involved in immune-related (tetraspanin and C-type lectins) and detoxification pathways as significantly up-regulated in the hemocytes of Cry1F treated ACB-FR. High-throughput in vitro assays revealed the binding affinity of Cry1F with the tetraspanin and C-type lectin family proteins. We found significant activation of MAPKinase (ERK 1/2, p38α, and JNK 1/2) in the hemocytes of Cry1F treated ACB-FR. In testing plausible crosstalk between a tetraspanin (CD63) and downstream MAPK signaling, we knocked down CD63 expression by RNAi and detected an alteration in JNK 1/2 level but a significant increase in susceptibility of ACB-FR larvae to Cry1F toxin. Information from this study advances a change in knowledge on the cellular immune response to Cry intoxication and its potential role in resistance in a lepidopteran pest.


Assuntos
Bacillus thuringiensis , Animais , Humanos , Larva , Hemócitos , Zea mays , Lectinas Tipo C
10.
Toxins (Basel) ; 14(12)2022 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-36548720

RESUMO

The crystal protein Cry5B, a pore-forming protein produced by the soil bacterium Bacillus thuringiensis, has been demonstrated to have excellent anthelmintic activity. While a previous structure of the three-domain core region of Cry5B(112-698) had been reported, this structure lacked a key N-terminal extension critical to function. Here we report the structure of Cry5B(27-698) containing this N-terminal extension. This new structure adopts a distinct quaternary structure compared to the previous Cry5B(112-698) structure, and also exhibits a change in the conformation of residues 112-140 involved in linking the N-terminal extension to the three-domain core by forming a random coil and an extended α-helix. A role for the N-terminal extension is suggested based on a computational model of the tetramer with the conformation of residues 112-140 in its alternate α-helix conformation. Finally, based on the Cry5B(27-698) structure, site-directed mutagenesis studies were performed on Tyr495, which revealed that having an aromatic group or bulky group at this residue 495 is important for Cry5B toxicity.


Assuntos
Bacillus thuringiensis , Bacillus thuringiensis/metabolismo , Endotoxinas/metabolismo , Toxinas de Bacillus thuringiensis/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Proteínas Hemolisinas/metabolismo
11.
Artigo em Inglês | MEDLINE | ID: mdl-36554372

RESUMO

The use of microbial insecticides and their toxins in biological control and transgenic plants has increased their presence in the environment. Although they are natural products, the main concerns are related to the potential impacts on the environment and human health. Several assays have been performed worldwide to investigate the toxicity or adverse effects of these microbial products or their individual toxins. This overview examines the published data concerning the knowledge obtained about the ecotoxicity and environmental risks of these natural pesticides. The data presented show that many results are difficult to compare due to the diversity of measurement units used in the different research data. Even so, the products and toxins tested present low toxicity and low risk when compared to the concentrations used for pesticide purposes. Complementary studies should be carried out to assess possible effects on human health.


Assuntos
Bacillus thuringiensis , Produtos Agrícolas , Inseticidas , Praguicidas , Plantas Geneticamente Modificadas , Bacillus thuringiensis/genética , Proteínas de Bactérias/genética , Produtos Agrícolas/toxicidade , Ecotoxicologia , Endotoxinas , Proteínas Hemolisinas/toxicidade , Inseticidas/toxicidade , Controle Biológico de Vetores/métodos , Praguicidas/toxicidade , Plantas Geneticamente Modificadas/toxicidade
12.
J Econ Entomol ; 115(6): 1752-1760, 2022 12 14.
Artigo em Inglês | MEDLINE | ID: mdl-36515105

RESUMO

The fall armyworm, Spodoptera frugiperda, is an economically important pest of corn, cotton, and soybean, and a major target of transgenic crops expressing Bacillus thuringiensis (Bt) proteins. In recent years, this insect has invaded most countries in Africa, Southeastern Asia, and Oceania, posing a great threat to food security. Successful use of Bt crops in the U.S. indicates that Bt technology can be an effective tool for management of S. frugiperda in other countries. Evolution of insect resistance is the primary threat to the long-term efficacy of Bt technology. There are many factors that may affect the rate of evolution of insect resistance to Bt crops, which include initial resistance allele frequency, the dose of Bt protein in Bt crops, cross-resistance, complete/incomplete resistance, and fitness costs associated with resistance. Currently, the high dose/refuge and gene-pyramiding approaches are the two main IRM strategies used in the U.S. to combat evolution of insect resistance. In this paper, we review research on resistance of S. frugiperda to Cry1, Cry2, and Vip3Aa proteins. Specifically, we discuss the resistance allele frequencies of S. frugiperda to these three proteins in the field, the genetic basis of resistance, the patterns of cross-resistance, and the fitness costs associated with resistance. Experience and knowledge gained from these studies provide valuable information for the successful use of Bt crop technology for control of S. frugiperda worldwide.


Assuntos
Bacillus thuringiensis , Animais , Bacillus thuringiensis/genética , Spodoptera/genética , Zea mays/genética , Zea mays/metabolismo , Plantas Geneticamente Modificadas/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/farmacologia , Proteínas de Bactérias/metabolismo , Proteínas Hemolisinas/farmacologia , Proteínas Hemolisinas/metabolismo , Gossypium/genética , Produtos Agrícolas , América , Resistência a Inseticidas/genética , Endotoxinas/farmacologia
13.
Int J Mol Sci ; 23(23)2022 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-36499184

RESUMO

The use of insect-resistant transgenic crops producing Bacillus thuringiensis protein Cry toxins (Bt) to control caterpillars is wide-spread. Development of a mechanism to prevent Bt from reaching its target site in the digestive system could result in Bt resistance and resistance to other insecticides active per os. Increased feeding rates by increasing temperature in tobacco budworms, Chloridea virescens, and bollworms, Helicoverpa zea, decreased Bt Cry1Ac susceptibility and mortality. The same was found in C. virescens for Bollgard II plant extract containing Bt Cry1Ac and Cry2Ab2 toxins. Furthermore, H. zea from the same inbred laboratory colony that fed faster independent of temperature manipulation were less susceptible to Bt intoxication. A laboratory derived C. virescens Bt resistant strain demonstrated a higher feeding rate on non-Bt artificial diet than the parental, Bt susceptible strain. A laboratory-reared Bt resistant fall armyworm, Spodoptera frugiperda, strain also fed faster on non-Bt diet compared to Bt susceptible caterpillars of the same species, both originally collected from corn. The studies in toto and the literature reviewed support the hypothesis that increased feeding rate is a behavioral mechanism for reducing caterpillar susceptibility to Bt. Its possible role in resistance needs further study.


Assuntos
Bacillus thuringiensis , Mariposas , Animais , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/farmacologia , Proteínas Hemolisinas/metabolismo , Endotoxinas/genética , Endotoxinas/metabolismo , Controle Biológico de Vetores , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Toxinas de Bacillus thuringiensis , Mariposas/genética , Bacillus thuringiensis/genética , Bacillus thuringiensis/metabolismo , Resistência a Inseticidas/genética , Gossypium/metabolismo , Larva/metabolismo
14.
Toxins (Basel) ; 14(12)2022 12 03.
Artigo em Inglês | MEDLINE | ID: mdl-36548749

RESUMO

Spodoptera frugiperda (fall armyworm, FAW) is one of the most devastating insect pests to corn and soybean production in the Americas and is rapidly expanding its range worldwide. It is known to be hard to control either by chemical insecticide applications or by GM. Although the use of GM traits can be an effective way to control this pest, it is very rare to find native insecticidal proteins that provide the necessary level of FAW control in crop fields where FAW pressure and damage are high. Insecticidal Cry proteins sourced from Bacillus thuringiensis have been heavily utilized in the development of crops with GM traits; however, it is increasingly difficult to identify Cry proteins with unique modes of action. Protein engineering via a phylogenetically guided Cry protein domain swapping approach enabled us to discover novel chimeric Cry proteins engineered from inactive parent sequences. Some of these chimeras show excellent efficacy against key biotypes of FAW from Brazil and North America. In this study, we characterized a Cry-based chimera eCry1Gb.1Ig that is a very potent FAW toxin. eCry1Gb.1Ig showed high efficacy against multiple FAW strains that are resistant to various traits, including Cry1Fa, Vip3Aa and Cry1A.105/Cry2Ab. These results clearly indicate that the FAW strains resistant to Cry1Fa, Vip3Aa or Cry1A.105/Cry2Ab demonstrate no cross-resistance to eCry1Gb.1Ig and strongly suggest that eCry1Gb.1Ig acts through a novel mode of action compared to the existing traits. In addition to its FAW activity, eCry1Gb.1Ig has also been shown to control Chrysodeixis includens (soybean looper, SBL) and Anticarsia gemmatalis (velvetbean caterpillar, VBC), which are significant pests of soybean. When eCry1Gb.1Ig was introduced into corn and soybean crops, transgenic events showed strong efficacy against FAW, SBL and VBC, but no adverse plant phenotypes. This suggests that the in planta expression of the eCry1Gb.1Ig protein does not compromise plant growth or reproduction and can protect plants from FAW-related damage. Therefore, this valuable discovery will provide a differentiating FAW control trait that will give growers another tool to help them reduce yield loss due to FAW.


Assuntos
Bacillus thuringiensis , Inseticidas , Mariposas , Animais , Spodoptera , Endotoxinas/genética , Endotoxinas/farmacologia , Endotoxinas/metabolismo , Toxinas de Bacillus thuringiensis/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/farmacologia , Proteínas de Bactérias/metabolismo , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/farmacologia , Proteínas Hemolisinas/metabolismo , Mariposas/genética , Inseticidas/farmacologia , Inseticidas/metabolismo , Bacillus thuringiensis/genética , Produtos Agrícolas/metabolismo , Zea mays/genética , Zea mays/metabolismo , Soja/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo
15.
Toxins (Basel) ; 14(12)2022 12 08.
Artigo em Inglês | MEDLINE | ID: mdl-36548760

RESUMO

Tpp80Aa1 from Bacillus thuringiensis is a Toxin_10 family protein (Tpp) with reported action against Culex mosquitoes. Here, we demonstrate an expanded target range, showing Tpp80Aa1 is also active against the larvae of Anopheles gambiae and Aedes aegypti mosquitoes. We report the first crystal structure of Tpp80Aa1 at a resolution of 1.8 Å, which shows Tpp80Aa1 consists of two domains: an N-terminal ß-trefoil domain resembling a ricin B lectin and a C-terminal putative pore-forming domain sharing structural similarity with the aerolysin family. Similar to other Tpp family members, we observe Tpp80Aa1 binds to the mosquito midgut, specifically the posterior midgut and the gastric caecum. We also identify that Tpp80Aa1 can interact with galactose-containing glycolipids and galactose, and this interaction is critical for exerting full insecticidal action against mosquito target cell lines.


Assuntos
Aedes , Bacillus thuringiensis , Culex , Inseticidas , Animais , Bacillus thuringiensis/metabolismo , Galactose/metabolismo , Aedes/metabolismo , Inseticidas/química , Culex/metabolismo , Proteínas de Bactérias/metabolismo , Larva/metabolismo , Endotoxinas/metabolismo , Proteínas Hemolisinas/metabolismo
16.
Molecules ; 27(21)2022 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-36364090

RESUMO

Parasporin 2 has cytotoxic effects against numerous colon cancer cell lines, making it a viable alternative to traditional treatments. However, its mechanism of action and receptors remain unknown. In this study, site-directed mutagenesis was used to obtain PS2Aa1 mutants with variation in domain I at positions 256 and 257. Variants 015, 002, 3-3, 3-35, and 3-45 presented G256A, G256E, G257A, G257V, and G257E substitutions, respectively. Cytotoxicity tests were performed for the cell viability of cell lines SW480, SW620, and CaCo-2. Mutants 3-3, 3-35, and 3-45 efficiently killed the cell lines. It was found that the activated forms of caspase-3 and PARP were in higher abundance as well as increased production of γH2AX when 3-35 was used to treat CaCo-2 and SW480. To assess possible membrane-binding receptors involved in the interaction, an APN receptor blocking assay showed reduced activity of some parasporins. Hence, we performed molecular docking and molecular dynamics simulations to analyze the stability of possible interactions and identify the residues that could be involved in the protein-protein interaction of PS2Aa1 and APN. We found that residues 256 and 257 facilitate the interaction. Parasporin 3-35 is promising because it has higher cytotoxicity than PS2Aa1.


Assuntos
Antineoplásicos , Bacillus thuringiensis , Neoplasias Colorretais , Humanos , Bacillus thuringiensis/metabolismo , Simulação de Acoplamento Molecular , Células CACO-2 , Antineoplásicos/farmacologia , Neoplasias Colorretais/tratamento farmacológico , Neoplasias Colorretais/genética , Linhagem Celular Tumoral , Apoptose
17.
Commun Biol ; 5(1): 1286, 2022 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-36434275

RESUMO

Bacillus thuringiensis serovar israelensis is the most widely used biopesticide against insects, including vectors of animal and human diseases. Among several extrachromosomal elements, this endospore-forming entomopathogen harbors two bacteriophages: a linear DNA replicon named GIL01 that does not integrate into the chromosome during lysogeny and a circular-jumbo prophage known as pBtic235. Here, we show that GIL01 hinders the induction of cohabiting prophage pBtic235. The GIL01-encoded small protein, gp7, which interacts with the host LexA repressor, is a global transcription regulator and represses the induction of pBtic235 after DNA damage to presumably allow GIL01 to multiply first. In a complex with host LexA in stressed cells, gp7 down-regulates the expression of more than 250 host and pBtic235 genes, many of which are involved in the cellular functions of genome maintenance, cell-wall transport, and membrane and protein stability. We show that gp7 homologs that are found exclusively in bacteriophages act in a similar fashion to enhance LexA's binding to DNA, while likely also affecting host gene expression. Our results provide evidence that GIL01 influences both its host and its co-resident bacteriophage.


Assuntos
Bacillus thuringiensis , Bacteriófagos , Animais , Humanos , Bacillus thuringiensis/genética , Bacillus thuringiensis/metabolismo , Bacteriófagos/genética , Sorogrupo , Lisogenia/genética , DNA/metabolismo
18.
J Microbiol Methods ; 203: 106622, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36384173

RESUMO

Although electroporation technique has been mostly used to transform Bacillus thuringiensis (Bt), this method is not readily applicable to strains other than the one for which it was optimized. Polyethylenimine (PEI) is a golden standard non-viral vector that interacts with plasmids to form compact polymeric nanoparticles (PNPs) via electrostatic interactions. This PNPs system is very attractive because they are easily prepared, able to carry large nucleic acid constructs, and show low toxicity. In this study, PEI/pBTdsSBV-VP1 PNPs were successfully prepared at various N/P ratios which is positively-chargeable polymer amine (N = nitrogen) groups to negatively-charged nucleic acid phosphate (P) groups, and the internalization of the complexes into Bt 4Q7 was confirmed by confocal laser scanning microscopy. The PEI-mediated transformation showed similar efficiency comparable to that of electroporation method, suggesting that the method of PNPs will be an effective alternative for transformation of Bt strains.


Assuntos
Bacillus thuringiensis , Nanopartículas , Ácidos Nucleicos , Polietilenoimina , Bacillus thuringiensis/genética , Polímeros , Plasmídeos/genética , DNA
19.
Parasit Vectors ; 15(1): 423, 2022 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-36369170

RESUMO

BACKGROUND: The common house mosquito Culex pipiens is known to be a major vector for West Nile virus. In order to decrease risks of West Nile virus outbreaks in Europe, insecticides and the bio-larvicide Bacillus thuringiensis israelensis (Bti) are commonly used for vector control. Alarmingly, insecticide resistance has been reported in Cx. pipiens populations from Southern Europe and several countries neighbouring Europe. For Central and Northern Europe, however, the phenotypic insecticide resistance status of Cx. pipiens has not yet been investigated. METHODS: A literature review was performed to assess the geographical distribution of insecticide resistance in Cx. pipiens. To fill the gap of knowledge for Central and Northern Europe, WHO susceptibility tests with permethrin, deltamethrin, malathion, bendiocarb and DDT and a larval toxicity test with Bti were performed with a Cx. pipiens population from Belgium, a country in Central Europe. RESULTS: This research provides the first evidence of widespread phenotypic insecticide resistance in Cx. pipiens. In general, Cx. pipiens developed resistance against multiple insecticides in several countries. Another Cx. pipiens population from Belgium was tested and showed insecticide resistance against deltamethrin, permethrin, DDT and possibly against bendiocarb. The bio-larvicide Bti caused lower mortality than reported for other Cx. pipiens populations in the literature. CONCLUSIONS: These results indicate the urgent need for insecticide resistance monitoring against commonly used adulticides and larvicides in Europe, for the translation of knowledge gained regarding the limited efficiency and availability of insecticide into EU legislation and the need for innovative non-chemical vector control tools in order to counter the widespread insecticide resistance in Culex populations.


Assuntos
Bacillus thuringiensis , Culex , Inseticidas , Vírus do Nilo Ocidental , Animais , Resistência a Inseticidas , Inseticidas/farmacologia , Permetrina , DDT , Controle de Mosquitos , Mosquitos Vetores
20.
Malar J ; 21(1): 328, 2022 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-36376966

RESUMO

BACKGROUND: In 2017, several new housing districts were constructed on Bioko Island, Equatorial Guinea. This case study assessed the impact construction projects had on mosquito larval habitats and the effectiveness of larval source management in reducing malaria vector density within the surrounding area. METHODS: Anopheline larval presence was assessed at 11 new construction sites by the proportion of larval habitats containing Anopheline pupae and late instar larval stages. Bacillus thuringiensis israelensis (Bti) larvicide was applied weekly to nine locations for 30 weeks, while two locations received no larvicide and acted as controls. Adult mosquito density was monitored via human landing collections in adjacent communities of six construction sites, including the two control sites. RESULTS: The sites that received Bti had significantly lower observation rates of both pupae (3.2% vs. 18.0%; p < 0.001) and late instar Anopheles spp. mosquitoes (14.1 vs. 43.6%; p < 0.001) compared to the two untreated sites. Anopheles spp. accounted for 67% of mosquitoes collected with human landing collections and were captured at significantly lower levels in communities adjacent to treated construction sites compared to untreated sites (p < 0.001), with an estimated 38% reduction in human biting rate (IRR: 0.62, 95% CI IRR: 0.55, 0.69). Seven months after the start of the study, untreated sites were treated due to ethical concerns given results from treatment sties, necessitating immediate Bti application. The following week, the number of habitats, the proportion of larval sites with Anopheles spp. pupae, late instars, and adult biting rates in adjacent communities to these sites all decreased to comparable levels across all sites. CONCLUSION: Findings suggest larval source management represents an effective intervention to suppress mosquito populations during infrastructure development. Incorporating larval source management into ongoing and planned construction initiatives represents an opportunity to fine tune vector control in response to anthropogenetic changes. Ideally, this should become standard practice in malaria-endemic regions in order to reduce viable mosquito habitats that are common by-products of construction.


Assuntos
Anopheles , Bacillus thuringiensis , Malária , Animais , Humanos , Anopheles/fisiologia , Malária/epidemiologia , Controle de Mosquitos/métodos , Larva , Reforma Urbana , Mosquitos Vetores , Pupa , Ecossistema
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