Discovery of crystalline inclusions in Bacillus licheniformis that resemble parasporal crystals of Bacillus thuringiensis

2007 ◽  
Vol 53 (9) ◽  
pp. 1111-1115 ◽  
Author(s):  
Ming Yan ◽  
Michael H. Roehrl ◽  
Julia Y. Wang
Keyword(s):  
Bacillus Thuringiensis ◽  
Bacillus Licheniformis ◽  
Rrna Gene ◽  
Bacterial Cells ◽  
Insecticidal Toxin ◽  
Crystalline Inclusions ◽  
Large Size ◽  
Protein Toxins ◽  
Bacterium Bacillus ◽  
Parasporal Crystals

Crystalline inclusions were discovered in stationary and sporulating cells of the spore-forming bacterium Bacillus licheniformis ATCC 9945a. As detected by electron microscopy, dying or sporulating bacterial cells contain a single crystal of strikingly large size. The crystals in sporulating cells are located next to nascent spores and can be several times larger than the spores. Morphologically, most crystals are rhomboid with uniformly spaced grids. These newly discovered crystalline inclusions of B. licheniformis closely resemble parasporal crystals of Bacillus thuringiensis that are formed by insecticidal toxin proteins and used widely as biopesticides. The taxonomic identity of this strain was verified by its 16S rRNA gene sequence and its fatty acid profile. The finding of crystal proteins in B. licheniformis may lead to the discovery of new protein toxins and may expand our pool of biopesticides.

Molecular Barcoding Based 16S rRNA Gene of Thermophilic Bacteria from Vulcanic Sites, Linow Lake, Tomohon

2019 ◽  
Vol 967 ◽  
pp. 83-92 ◽  
Author(s):  
Simanjuntak Suddin ◽  
Yermia Semuel Mokosuli ◽  
Welerubun Marcelina ◽  
Naharia Orbanus ◽  
Kapahang Ardi
Keyword(s):  
Bacillus Thuringiensis ◽  
16S Rrna ◽  
Bacillus Licheniformis ◽  
Thermophilic Bacteria ◽  
Extraction Procedure ◽  
Compatible Solutes ◽  
Rrna Gene ◽  
Accession Number ◽  
16S Rna ◽  
Thuringiensis Strain

Thermophilic bacteria live at temperatures above 450 C. Many investigations focused on their potential as sources of highly active enzymes ‘termostable enzyme’ and other products such as antibiotics and compatible solutes. Lake Linow is an active volcanic lake located in Tomohon City, North Sulawesi, Indonesia. Lake Linow becomes the habitat of thermophilic bacteria. A study has been conducted to obtain isolates of thermophilic bacteria and to identifikasi berdasarkan gen 16 s RNA. Bacterial DNA extraction procedure using the Presto TM Mini gDNA Bacteria Kit Geneaid protocol, with modifications. Amplification of 16s RNA gene using PCR method. Visualization of 16 s RNA amplicon genes with automatic electrophoresis capiler Qiaxel, Qiagen. Sequencing was carried out using Singapore's First BASE Sequencing service. The results showed that IL2 isolates and IL3 isolates could live up to 700C. Alignment analysis results using NCAST BLBI IL2 isolates showed 99% similarity with Bacillus thuringiensis strain H2682 (accession number CP009720.1). While isolate of IL3 thermophilic bacteria showed 94% similarity with Bacillus licheniformis strain 14DA11 (accession number CP023168.1). The results of phylogeny reconstruction with neighbor joining method, gene sequence 16S rRNA isolate IL2 showed the closest relation with Bacillus thuringiensis strain HD1011 (accession number CP009335.1). While IL3 isolate showed the closest relation with Bacillus licheniformis strain 14DA11 (accession number CP023168.1).

scholarly journals Isolation of Bacillus thuringiensis strains from Saudi Arabia soil and study of their potential efficiency against the lepidopteran pest Ephestia kuehniella

2021 ◽  
Vol 12 (2) ◽  
pp. 238-245
Author(s):  
Fidaa Ibrahim Kallaf ◽  
Hanen Boukedi ◽  
Dalel Daâssi ◽  
Lobna Abdelkefi-Mesrati
Keyword(s):  
Saudi Arabia ◽  
Bacillus Thuringiensis ◽  
Insect Pests ◽  
Ephestia Kuehniella ◽  
Environment Friendly ◽  
Insecticidal Proteins ◽  
Insect Pathogens ◽  
Bacterium Bacillus ◽  
Parasporal Crystals ◽  
Lepidopteran Pest

Insect pests represent a major threat to food crops and human health, and therefore have to be combated in several ways, including chemical methods. However, researchers demonstrated that these molecules are dangerous for the farmers, consumers and the environment in general. For this reason, scientists permanently searched environment friendly alternatives such as the use of the bacterium Bacillus thuringiensis classified as one of the best insect pathogens. This microorganism is known by its ability to produce two types of insecticidal proteins, Vegetative insecticidal proteins (Vip) and delta-endotoxins produced during vegetative and sporulation stages of growth, respectively. In the present study, 15 B. thuringiensis strains were isolated from soil collected from different regions in Saudi Arabia (Al Baha, Jeddah, Khulis and Yanbu). B. thuringiensis isolates were then classified according to the shape of their parasporal crystals identified under microscope and proteins content of these crystals. Delta-endotoxins efficiency of the different isolates was investigated and promising strains were identified as very active. After 5 days-treatment, B. thuringiensis isolates 14 and 7 killed Ephestia kuehniella larvae with low LC50 of about 59.18 and 65.67 mg/cm2, respectively. The results described in the present study proved that the new B. thuringiensis isolates could be of a great interest in the control of lepidopteran pests by using their delta-endotoxins in bioinsecticide formulations.

scholarly journals The human cancer cell active toxin Cry41Aa from Bacillus thuringiensis acts like its insecticidal counterparts

2017 ◽  
Vol 474 (10) ◽  
pp. 1591-1602 ◽  
Author(s):  
Vidisha Krishnan ◽  
Barbara Domanska ◽  
Alicia Elhigazi ◽  
Fatai Afolabi ◽  
Michelle J. West ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Mechanism Of Action ◽  
Human Cell ◽  
Human Cancer ◽  
Expression System ◽  
Membrane Damage ◽  
Cell Swelling ◽  
Loop Region ◽  
Protein Toxins ◽  
Bacterium Bacillus

Understanding how certain protein toxins from the normally insecticidal bacterium Bacillus thuringiensis (Bt) target human cell lines has implications for both the risk assessment of products containing these toxins and potentially for cancer therapy. This understanding requires knowledge of whether the human cell active toxins work by the same mechanism as their insecticidal counterparts or by alternative ones. The Bt Cry41Aa (also known as Parasporin3) toxin is structurally related to the toxins synthesised by commercially produced transgenic insect-resistant plants, with the notable exception of an additional C-terminal β-trefoil ricin domain. To better understand its mechanism of action, we developed an efficient expression system for the toxin and created mutations in regions potentially involved in the toxic mechanism. Deletion of the ricin domain did not significantly affect the activity of the toxin against the human HepG2 cell line, suggesting that this region was not responsible for the mammalian specificity of Cry41Aa. Various biochemical assays suggested that unlike some other human cell active toxins from Bt Cry41Aa did not induce apoptosis, but that its mechanism of action was consistent with that of a pore-forming toxin. The toxin induced a rapid and significant decrease in metabolic activity. Adenosine triphosphate depletion, cell swelling and membrane damage were also observed. An exposed loop region believed to be involved in receptor binding of insecticidal Cry toxins was shown to be important for the activity of Cry41Aa against HepG2 cells.

scholarly journals No Evidence of an Impact on the Rhizosphere Diazotroph Community by the Expression of Bacillus thuringiensis Cry1Ab Toxin by Bt White Spruce

2007 ◽  
Vol 73 (20) ◽  
pp. 6577-6583 ◽  
Author(s):  
Josyanne Lamarche ◽  
Richard C. Hamelin
Keyword(s):  
Bacillus Thuringiensis ◽  
Bacterial Communities ◽  
White Spruce ◽  
Sequencing Analysis ◽  
Nitrogen Fixing ◽  
Cloning And Sequencing ◽  
Insecticidal Toxin ◽  
Soil Bacterial Communities ◽  
Cry1ab Toxin ◽  
Natural Stands

ABSTRACT Nitrogen fixation is one of the most important roles played by soil bacterial communities, as fixation supplies nitrogen to many ecosystems which are often N limited. As impacts on this functional group of bacteria might harm the ecosystem's health and reduce productivity, monitoring that particular group is important. Recently, a field trial with Bt white spruce, which constitutively expresses the Cry1Ab insecticidal toxin of Bacillus thuringiensis, was established. The Bt white spruce was shown to be resistant to spruce budworm. We investigated the possible impact of these genetically modified trees on soil nitrogen-fixing bacterial communities. The trial consisted of untransformed controls, GUS white spruce (transformed with the β-glucuronidase gene), and Bt/GUS white spruce (which constitutively expresses both the Cry1Ab toxin and β-glucuronidase) in a random design. Four years after planting, soil samples from the control and the two treatments from plantation as well as from two natural stands of white spruce were collected. Diazotroph diversity was assessed by extracting soil genomic DNA and amplifying a region of the nitrogenase reductase (nifH) gene, followed by cloning and sequencing. Analysis revealed that nitrogen-fixing communities did not differ significantly among the untransformed control, GUS white spruce, and Bt/GUS white spruce. Nevertheless, differences in diazotroph diversity were observed between white spruce trees from the plantation site and those from two natural stands, one of which grew only a few meters away from the plantation. We therefore conclude, in the absence of evidence that the presence of the B. thuringiensis cry1Ab gene had an effect on diazotroph communities, that either site and/or field preparation prior to planting seems to be more important in determining diazotroph community structure than the presence of Bt white spruce.

scholarly journals Draft genome sequence data of a chromium reducing bacterium, Bacillus licheniformis strain KNP

Data in Brief ◽  
2021 ◽  
Vol 34 ◽  
pp. 106640
Author(s):  
Pankaj Kumar Arora ◽  
Rupali Mishra ◽  
Rishabh Anand Omar ◽  
Raj Shekhar Saroj ◽  
Alok Srivastava ◽  
...  
Keyword(s):  
Genome Sequence ◽  
Bacillus Licheniformis ◽  
Sequence Data ◽  
Draft Genome ◽  
Draft Genome Sequence ◽  
Genome Sequence Data ◽  
Bacterium Bacillus

Function of the 28,000 and 66,000 mol. wt protein toxins of Bacillus thuringiensis

Toxicon ◽  
1997 ◽  
Vol 35 (4) ◽  
pp. 482
Author(s):  
Y. Nitzan ◽  
R. Cahan ◽  
I. Pechatnikov
Keyword(s):  
Bacillus Thuringiensis ◽  
Protein Toxins

scholarly journals THE USE OF AGENTS BASED ON THE BACTERIUM BACILLUS THURINGIENSIS VAR. ISRAELENSIS FOR MOSQUITO CONTROL

2019 ◽  
Vol 98 (8) ◽  
pp. 893-896
Author(s):  
Svetlana A. Roslavtseva
Keyword(s):  
Bacillus Thuringiensis ◽  
Aedes Aegypti ◽  
Synergistic Effect ◽  
Mosquito Control ◽  
Mosquito Species ◽  
Bacillus Sphaericus ◽  
Development Of Resistance ◽  
Blood Sucking ◽  
Low Concentrations ◽  
Bacterium Bacillus

Mosquito control is necessary to improve the epidemic and, consequently, the sanitary and hygienic situation in human settlements. At the same time, the safest and more environmentally friendly way of controlling is not the fight against adult mosquitoes, but the treatment of reservoirs with microbiological larvicides based on entomopathogenic, aerobic, spore-forming, saprophytic bacteria Bacillus thuringiensis (de Barjac) (Bti). A new serotype of the bacterium B. thuringiensis was found in Israel in the Negev desert. This serotype being more active against larvae of blood-sucking and non-blood-sucking mosquitoes and midges than previously known serotypes, was named israelensis. Bti endotoxin is a typical insecticide with intestinal type of action for different mosquito species. For example, Bti H14 is highly insecticidal to the larvae of Aedes aegypti and Ae. albopictus at very low concentrations. The parasporal body (endotoxin crystal), a crystalline protein consisted of four main polypeptides and two minor polypeptides, possesses of a larvicidal action. Larvicidal activity is associated with a synergistic effect in a combination of four polypeptides. The possibility of development of resistance to products based on Bti and Bacillus sphaericus in populations of mosquitoes (Culicidae) was investigated. The use of domestic microbiological formulations based on Bti («Baktitsid», «Larviol-pasta», and «Antinat») was shown an eradication the larvae of bloodsucking mosquitoes and midges to be possible and rational, since they are not generated resistant populations of mosquitoes. This is confirmed by more than 30 years of the use of such formulations.

scholarly journals Titanium Dioxide Nanoparticles Induce Inhibitory Effects against Planktonic Cells and Biofilms of Human Oral Cavity Isolates of Rothia mucilaginosa, Georgenia sp. and Staphylococcus saprophyticus

Pharmaceutics ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 1564
Author(s):  
Saher Fatima ◽  
Khursheed Ali ◽  
Bilal Ahmed ◽  
Abdulaziz A. Al Kheraif ◽  
Asad Syed ◽  
...  
Keyword(s):  
Titanium Dioxide Nanoparticles ◽  
Anatase Phase ◽  
Spherical Shape ◽  
Rrna Gene ◽  
Dependent Manner ◽  
Bacterial Cells ◽  
Gram Positive ◽  
Concentration Dependent Manner ◽  
Staphylococcus Saprophyticus ◽  
Rothia Mucilaginosa

Multi-drug resistant (MDR) bacterial cells embedded in biofilm matrices can lead to the development of chronic cariogenesis. Here, we isolated and identified three Gram-positive MDR oral cocci, (1) SJM-04, (2) SJM-38, and (3) SJM-65, and characterized them morphologically, biochemically, and by 16S rRNA gene-based phylogenetic analysis as Georgenia sp., Staphylococcus saprophyticus, and Rothia mucilaginosa, respectively. These three oral isolates exhibited antibiotic-resistance against nalidixic acid, tetracycline, cefuroxime, methicillin, and ceftazidime. Furthermore, these Gram positive MDR oral cocci showed significant (p < 0.05) variations in their biofilm forming ability under different physicochemical conditions, that is, at temperatures of 28, 30, and 42 °C, pH of 6.4, 7.4, and 8.4, and NaCl concentrations from 200 to 1000 µg/mL. Exposure of oral isolates to TiO2NPs (14.7 nm) significantly (p < 0.05) reduced planktonic cell viability and biofilm formation in a concentration-dependent manner, which was confirmed by observing biofilm architecture by scanning electron microscopy (SEM) and optical microscopy. Overall, these results have important implications for the use of tetragonal anatase phase TiO2NPs (size range 5–25 nm, crystalline size 13.7 nm, and spherical shape) as an oral antibiofilm agent against Gram positive cocci infections. We suggest that TiO2NPs pave the way for further applications in oral mouthwash formulations and antibiofilm dental coatings.

scholarly journals In Vivo Entombment of Bacteria and Fungi during Calcium Oxalate, Brushite and Struvite Urolithiasis

Kidney360 ◽  
2020 ◽  
pp. 10.34067/KID.0006942020
Author(s):  
Jessica J. Saw ◽  
Mayandi Sivaguru ◽  
Elena M. Wilson ◽  
Yiran Dong ◽  
Robert A. Sanford ◽  
...  
Keyword(s):  
Calcium Oxalate ◽  
Stone Formation ◽  
Human Kidney ◽  
Rrna Gene ◽  
Bacterial Cells ◽  
Stone Formers ◽  
Bacteria And Fungi ◽  
Struvite Stones

Background: Human kidney stones form via repeated events of mineral precipitation, partial dissolution and reprecipitation, which are directly analogous to similar processes in other natural and man-made environments where resident microbiomes strongly influence biomineralization. High-resolution microscopy and high-fidelity metagenomic (microscopy-to-omics) analyses, applicable to all forms of biomineralization, have been applied to assemble definitive evidence of in vivo microbiome entombment during urolithiasis. Methods: Stone fragments were collected from a randomly chosen cohort of 20 patients using standard percutaneous nephrolithotomy (PCNL). Fourier transform infrared (FTIR) spectroscopy indicated that 18 of these patients were calcium oxalate (CaOx) stone formers, while one patient each formed brushite and struvite stones. This apportionment is consistent with global stone mineralogy distributions. Stone fragments from 7 of these 20 patients (5 CaOx, 1 brushite and 1 struvite) were thin sectioned and analyzed using brightfield (BF), polarization (POL), confocal, superresolution autofluorescence (SRAF) and Raman techniques. DNA from remaining fragments, grouped according to each of the 20 patients, were analyzed with amplicon sequencing of 16S rRNA gene sequences (V1-V3, V3-V5) and internal transcribed spacer (ITS1, ITS2) regions. Results: Bulk entombed DNA was sequenced from stone fragments in 11 of the 18 CaOx patients, as well as the brushite and struvite patients. These analyses confirmed the presence of an entombed low-diversity community of bacteria and fungi, including Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria, and Aspergillus niger. Bacterial cells ~1  µm in diameter were also optically observed to be entombed and well-preserved in amorphous hydroxyapatite spherules and fans of needle-like crystals of brushite and struvite. Conclusions: These results indicate a microbiome is entombed during in vivo CaOx stone formation. Similar processes are implied for brushite and struvite stones. This evidence lays the groundwork for future in vitro and in vivo experimentation to determine how the microbiome may actively and/or passively influence kidney stone biomineralization.

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