Species Of Genus Bacillus Research Articles

Diversity and Prevalence of Indigenous Soil Bacillus spp. in the Major Cabbage Growing Agroecological Zones of Uganda

Different species of genus Bacillus have been reported from different environments of the world. They are reported to play a role of soil fertility improvement, plant growth promotion and disease and pest management. Most of these reports on Bacillus species are from studies conducted outside Uganda and therefore information on the prevalence and diversity of bacillus species in Ugandan soils is scanty. This study aimed at determining the prevalence and diversity of Bacillus spp. isolated from the cabbage rhizosphere in the four major cabbage-growing agroecological zones of Uganda. The experiment was conducted in a laboratory at the College of Veterinary Medicine, Animal Resources and Biosafety Makerere University for morphological and biochemical identification of the Bacillus bacteria. DNA extraction and PCR were conducted at the College of Natural Resources Makerere University while sequencing was done at Macrogen laboratories in Korea and Inqaba Biotec in South Africa. Morphological, biochemical and genomic analyses revealed five Bacillus spp. (22 Bacillus strains) grouped as B. cereus, B. mycoides, B. thuringiensis, B. megaterium and B. bingmayongensis. B. cereus and B. megaterium were the most dominant and widely spread Bacillus spp. A phylogenetic tree indicated three major clads, showing that B. thuringiensis was closely related to B. cereus while B.bingmayongesis was closely related to B.megaterium. The B. mycoides were closely related to some B. cereus strains and B. bingmayongensis. The phylogenetic tree further showed that some Bacillus strains of the same species were distantly related. It was therefore concluded that most abundant and prevalent Bacillus spp. in Ugandan soils were B. cereus and B. megaterium. The presence and abundance of these bacillus species in the Ugandan soil presents an opportunity for soil scientists to innovatively manipulate them for use as biofertilizers and biopesticides for crop production and management. Such innovations would reduce the reliance of farmers on synthetic fertilizers that are pollutants to the environment and unhealthy to the users and consumers

Screening and characterization of biocontrol bacteria isolated from Ageratum conyzoides against Collectotrichum fructicola causing Chinese plum (Prunus salicina Lindl.) anthracnose.

Chinese plum (Prunus salicina Lindl.) is a nutritionally and economically important stone fruit widely grown around the world. Anthracnose, caused by Collectotrichum spp., is one of the primary biotic stress factors limiting plum production. Medicinal plants may harbor rhizospheric or endophytic microorganisms that produce bioactive metabolites that can be used as anthracnose biocontrol agents. Here, 27 bacterial isolates from the medicinal plant A. conyzoides with diverse antagonistic activities against C. fructicola were screened. Based on morphological, physiological, biochemical, and molecular characterization, 25 of these isolates belong to different species of genus Bacillus, one to Pseudomonas monsensis, and one more to Microbacterium phyllosphaerae. Eight representative strains showed high biocontrol efficacy against plum anthracnose in a pot experiment. In addition, several Bacillus isolates showed a broad spectrum of inhibitory activity against a variety of fungal phytopathogens. Analysis of the volatile organic compound profile of these eight representative strains revealed a total of 47 compounds, most of which were ketones, while the others included alkanes, alkenes, alcohols, pyrazines, and phenols. Overall, this study confirmed the potential value of eight bacterial isolates for development as anthracnose biocontrol agents.

Bacillus fonticola sp. nov., isolated from deep sea cold seep sediment.

In this study, we report a novel Gram-positive bacterium, designated as strain CS13T, isolated from deep-sea sediment collected in the cold seep area of the South China Sea. Growth of strain CS13T occurred at 16-37°C (optimum 25-28°C), pH 7.0-9.0 (optimum, 7.0), and 0-8% (w/v) NaCl (optimum, 2-3%). Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain CS13T belonged to the genus Bacillus. The closest phylogenetic neighbors of strain CS13T are Bacillus carboniphilus JCM 9731T (96.0%), Bacillus pakistanensis NCCP-168T (95.7%) and Bacillus acidicola 105-2T (95.6%). The genomic DNA G + C content of strain CS13T is 43.7mol%. The principal respiratory quinone was menaquinone 7 (MK-7). The polar lipids of CS13T contained diphosphatidylglycerol, phosphatidylglycerol, phospholipid, and glycolipid. The major fatty acids of CS13T contained anteiso-C15:0, anteiso-C17:0, C16:0 and C18:0. Strain CS13T harboured meso-diaminopimelic acid as the diagnostic diamino acid. Phylogenetic, physiological, biochemical, and morphological analyses suggested that strain CS13T represents a novel species of genus Bacillus, and the name Bacillus fonticola sp. nov. is proposed for the type species CS13T (= CCTCC AB 2019194T = JCM 33663T).

Physiological and Molecular-genetic Characteristic of Bacteria Strains, Isolated from Mountain Spring and Mineral Waters in Plovdiv Region, Bulgaria

The present study aims to investigate the microbiological research of mineral and spring mountain waters in the region of Plovdiv, Bulgaria. In Bulgaria, there are mineral and mountain spring waters, which are not subjected to physicochemical and microbiological control by the Regional Health Inspectorate, yet they are the most widely used springs by the population as sources of drinking water. The methodology is including the determination of total number of mesophilic aerobic and facultative anaerobic bacteria. The results showed 11 rod shaped, Gram–positive and spore forming bacteria and 2 rod shaped, Gram–negative non-spore forming bacteria are isolated from spring and healing waters. The colonial and physiological characteristics of the isolated strains are defined. The biochemical profiles of the selected bacteria are determined with the system for rapid identification API 50 CH. The strains are identified by software processing of the obtained biochemical profiles with apiweb®. Furthermore, the tested strains are genetically identified by molecular-genetic methods – ARDRA analysis and sequencing of the gene for the 16S rRNA. The results obtained from the biochemical and molecular-genetic studies indicate that the selected strains belong to different species of genus Bacillus, Aeromonas, Klebsiella.
 The identification strains support for registration of water sources in Plovdiv region, Bulgaria according Decree No. 9.

Bacterial and Fungal Diversity Inside the Medieval Building Constructed with Sandstone Plates and Lime Mortar as an Example of the Microbial Colonization of a Nutrient-Limited Extreme Environment (Wawel Royal Castle, Krakow, Poland).

Biodeterioration is a serious threat to cultural heritage objects and buildings. The deterioration of a given material often incurs irreparable losses in terms of uniqueness and historical value. Hence preventive actions should be taken. One important challenge is to identify microbes involved in the biodeterioration process. In this study, we analyzed the microbial diversity of an ancient architectonical structure of the Rotunda of Sts. Felix and Adauctus, which is a part of the Wawel Royal Castle located in Krakow, Poland. The Rotunda is unavailable to tourists and could be treated as an extreme habitat due to the low content of nutrients coming either from sandstone plates bound with lime mortar or air movement. Microbial diversity was analyzed with the use of the high-throughput sequencing of marker genes corresponding to fragments of 16S rDNA (for Bacteria) and ITS2 (internal transcribed spacer 2) (for Fungi). The results showed that the microbial community adhered to wall surfaces is, to a large extent, endemic. Furthermore, alongside many microorganisms that could be destructive to masonry and mortar (e.g., Pseudomonas, Aspergillus), there were also bacteria, such as species of genera Bacillus, Paenisporosarcina, and Amycolatopsis, that can positively affect wall surface properties by reducing the damage caused by the presence of other microorganisms. We also showed that airborne microorganisms probably have little impact on the biodeterioration process as their abundance in the microbial community adhered to the ancient walls was very low.

Isolation and Molecular Characterization of Biosurfactant-Producing Bacterial Diversity of Fimkassar Oil Field, Pakistan

Oil-contaminated sites potentially harbor biosurfactant-producing microorganisms. The present study relates to isolation of biosurfactant-producing bacteria from Fimkassar oil field, Chakwal, Pakistan, and their molecular screening for two important genes viz. srfA and rhlB, responsible for production of surfactin and rhamnolipid biosurfactants, respectively. In total, 38 out of 79 different bacterial isolates showing growth on crude-oil-containing media were screened for biosurfactant production. Evidently, 34.2% ( $$n=13$$ ) of the isolates were found to inherit srfA gene, while 15.8% ( $$n=6$$ ) of the isolates contained rhlB gene. Subsequently, 16S ribosomal RNA sequence homology studies confirmed the gene-positive isolates to be the species of genera Bacillus, Brevundimonas, Alcaligenes, Pseudomonas, Serratia, Proteus and Stenotrophomonas. The unusual presence of genes in these microorganisms indicates the possibility of horizontal gene transfer and retention or presence of gene orthologs. Although all the gene-positive isolates showed biosurfactant production under submerged fermentative conditions, maximum production in terms of biosurfactant activities (i.e., $$E_{24} = 59.5\pm 4.0\%$$ ; $$\hbox {SFT} = 27.2\pm 1.1\, \hbox {mN/m}$$ ; $$\hbox {ODA} = 3.5\,\pm \,0.2\, \hbox {cm}$$ ) was revealed by Bacillus subtilis strain SWW1. Surfactin nature of biosurfactant produced was confirmed by thin-layer chromatography and Fourier transform infrared spectroscopy.

Evaluation of β-1,4-endoglucanases produced by bacilli isolated from paper and pulp mill effluents irrigated soil.

A total of 10 cellulase-producing bacteria were isolated from soil samples irrigated with paper and pulp mill effluents. The sequencing of 16S rRNA gene revealed that all isolates belonged to different species of genus Bacillus. Among the different isolates, B. subtilis IARI-SP-1 exhibited a high degree of β-1,4-endoglucanase (2.5 IU/ml), β-1,4-exoglucanase (0.8 IU/ml), and β-glucosidase (0.084 IU/ml) activity, followed by B. amyloliquefaciens IARI-SP-2. CMC was found to be the best carbon source for production of endo/exoglucanase and β-glucosidase. The β-1,4-endoglucanase gene was amplified from all isolates and their deduced amino acid sequences belonged to glycosyl hydrolase family 5. Among the domains of different isolates, the catalytic domains exhibited the highest homology of 93.7%, whereas the regions of signal, leader, linker, and carbohydrate-binding domain indicated low homology (73-74%). These variations in sequence homology are significant and could contribute to the structure and function of the enzyme.

Bacterial community compositions of tomato (Lycopersicum esculentum Mill.) seeds and plant growth promoting activity of ACC deaminase producing Bacillus subtilis (HYT-12-1) on tomato seedlings

Study of endophytic bacteria within plant seeds is very essential and meaningful on account of their heritability and versatility. This study investigated Bacillus bacterial communities within the seeds of four commercial tomato varieties, by 16S rRNA gene PCR-RFLP (restriction fragment length polymorphism). Phylogenetic analysis of 16S rRNA gene sequences indicated that the 22 representative isolates belonged to five species of genus Bacillus and the bacterial compositions showed remarkable differences among tomato varieties. Isolates exhibited multiple plant growth promoting (PGP) traits: 37 % of indole-3-acetic acid production; 37 % of phosphate solubilization; 24 % of siderophores production; 85 % of potential nitrogen fixation and 6 % of 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity. Isolate HYT-12-1 was shown to have highest ACC deaminase activity (112.02 nmol α-ketobutyrate mg⁻¹ protein h⁻¹) among the five ACC deamiase producing strains. 16S rRNA gene sequencing indicated that the isolate HYT-12-1 shared the highest sequence similarity (100 %) with B. subtilis. PGP experiments under gnotobiotic and greenhouse conditions revealed the ability of strain HYT-12-1 to enhance the growth of tomato seedlings. This is the first study to describe endophytic Bacillus communities within tomato seeds, and the results suggest that B. subtilis strain HYT-12-1 would have a great potential for industrial application as biofertilizer in the future.

Pathogenic Bacillus anthracis in the progressive gene losses and gains in adaptive evolution

BackgroundSequence mutations represent a driving force of adaptive evolution in bacterial pathogens. It is especially evident in reductive genome evolution where bacteria underwent lifestyles shifting from a free-living to a strictly intracellular or host-depending life. It resulted in loss-of-function mutations and/or the acquisition of virulence gene clusters. Bacillus anthracis shares a common soil bacterial ancestor with its closely related bacillus species but is the only obligate, causative agent of inhalation anthrax within the genus Bacillus. The anthrax-causing Bacillus anthracis experienced the similar lifestyle changes. We thus hypothesized that the bacterial pathogen would follow a compatible evolution path.ResultsIn this study, a cluster-based evolution scheme was devised to analyze genes that are gained by or lost from B. anthracis. The study detected gene losses/gains at two separate evolutionary stages. The stage I is when B. anthracis and its sister species within the Bacillus cereus group diverged from other species in genus Bacillus. The stage II is when B. anthracis differentiated from its two closest relatives: B. cereus and B. thuringiensis. Many genes gained at these stages are homologues of known pathogenic factors such those for internalin, B. anthracis-specific toxins and large groups of surface proteins and lipoproteins.ConclusionThe analysis presented here allowed us to portray a progressive evolutionary process during the lifestyle shift of B. anthracis, thus providing new insights into how B. anthracis had evolved and bore a promise of finding drug and vaccine targets for this strategically important pathogen.

Biodiversity and phylogenetic analysis of culturable bacteria indigenous to Khewra salt mine of Pakistan and their industrial importance

Culturable bacterial biodiversity and industrial importance of the isolates indigenous to Khewra salt mine, Pakistan was assessed. PCR Amplification of 16S rDNA of isolates was carried out by using universal primers FD1 and rP1and products were sequenced commercially. These gene sequences were compared with other gene sequences in the GenBank databases to find the closely related sequences. The alignment of these sequences with sequences available from GenBank database was carried out to construct a phylogenetic tree for these bacteria. These genes were deposited to GenBank and accession numbers were obtained. Most of the isolates belonged to different species of genus Bacillus, sharing 92-99% 16S rDNA identity with the respective type strain. Other isolates had close similarities with Escherichia coli, Staphylococcus arlettae and Staphylococcus gallinarum with 97%, 98% and 99% 16S rDNA similarity respectively. The abilities of isolates to produce industrial enzymes (amylase, carboxymethylcellulase, xylanase, cellulase and protease) were checked. All isolates were tested against starch, carboxymethylcellulose (CMC), xylane, cellulose, and casein degradation in plate assays. BPT-5, 11,18,19 and 25 indicated the production of copious amounts of carbohydrates and protein degrading enzymes. Based on this study it can be concluded that Khewra salt mine is populated with diverse bacterial groups, which are potential source of industrial enzymes for commercial applications.

Biodiversity and phylogenetic analysis of culturable bacteria indigenous to Khewra salt mine of pakistan and their industrial importance

Culturable bacterial biodiversity and industrial importance of the isolates indigenous to Khewra salt mine, Pakistan was assessed. PCR Amplification of 16S rDNA of isolates was carried out by using universal primers FD1 and rP1and products were sequenced commercially. These gene sequences were compared with other gene sequences in the GenBank databases to find the closely related sequences. The alignment of these sequences with sequences available from GenBank database was carried out to construct a phylogenetic tree for these bacteria. These genes were deposited to GenBank and accession numbers were obtained. Most of the isolates belonged to different species of genus Bacillus, sharing 92-99% 16S rDNA identity with the respective type strain. Other isolates had close similarities with Escherichia coli, Staphylococcus arlettae and Staphylococcus gallinarum with 97%, 98% and 99% 16S rDNA similarity respectively. The abilities of isolates to produce industrial enzymes (amylase, carboxymethylcellulase, xylanase, cellulase and protease) were checked. All isolates were tested against starch, carboxymethylcellulose (CMC), xylane, cellulose, and casein degradation in plate assays. BPT-5, 11,18,19 and 25 indicated the production of copious amounts of carbohydrates and protein degrading enzymes. Based on this study it can be concluded that Khewra salt mine is populated with diverse bacterial groups, which are potential source of industrial enzymes for commercial applications.

Number of Triplets in 16S rRNA Gene Related with Pathogenicity of Bacillus spp. and Clostridium spp.

The relation between the number of some trinucleotides in the sequence of 16S rRNA gene and pathogenicity of bacterial species from the genera of Bacillus and Clostridium was revealed. The species of genusBacillus , which are pathogenic for humans, mammals and insects, have an increased number of AAA and TAT triplets in 16S rRNA gene. Theoretically, these species, B. anthracis and B. cereus for example, may be detected in the specimen by the higher ratio of AAA plus TAT triplets to the number of GGG triplet. Species of genus Clostridium, which are pathogenic for humans and mammals, have a maximum ratio of AAA and TAT triplet numbers. This ratio was higher than 2.6 for pathogenic species and lower than 2.2 for saprophytic ones. These theoretical data may open a new way for detecting pathogenic bacteria through the determination of triplet numbers in the sequences of 16S rRNA or rRNA. However, the mechanism of evolutionary relation between the number of AAA and TAT triplets in the sequence of 16S rRNA gene and the pathogenicity of bacterial species is not known.

Rapid classification ofBacillus isolates using RAPD technique

This paper describes the typing of selectedBacillus strains from environmental origin and ATCC type strains. The objective of the study was to evaluate the usefulness of PCR-fingerprinting using Randomly Amplified Polymorphic DNA (RAPD) as a method for classifying members of the genusbacillus. Our experiments show that the levels of similarity of 69%∼90%, 40%∼60% were observed between the members of the same species and among the species of genusBacillus, respectively. In a dendrogram showing the positions of 28 strains, these strains were clearly seperated into distinct species, 13 of which are corresponding to type strains used in this study. These results suggest that RAPD analyses is a simple and powerful method for species identification and delimitation.

ISOLATION AND CHARACTERIZATION OF MICROORGANISMS INVOLVED IN THE FERMENTATION OF TRINIDAD'S CACAO BEANS

In order to isolate, identify and characterize the microflora of cacao beans before, during and after fermentation and locate possible sources contributing to microbial contamination, cacao beans from the Centeno and San Louis Estates in Trinidad were investigated. Prior to fermentation, the interior and exterior of the pods, hands of employees, utensils, dried pulp material of the sweatboxes and finally fruitflies (Drosophila melanogaster) were studied microbiologically. At Centeno Estate, beans were sampled at 5, 45 and 90 cm depths at 8‐hr intervals for the first 72 hr and every 12 hr thereafter for 7 days. Sampling at San Louis Estate was carried out at 24 hr intervals for the same period. The changes in microbial population of the beans sampled at Centeno Estate ranged from 1.48 × 105/g at 0 hr to 4.1 × 105/g at the completion of the fermentation, whereas, at San Louis Estate they ranged from 6.8 × 105/g to 9.2 × 105/g during the same period. Taxonomical studies of isolates obtained during the fermentation period revealed the identification of 44 microorganisms at both Estates. Yeasts Zymomonas mobilis and several species of lactic acid organisms dominated the flora during the early stages of fermentation. As the fermentation progressed, these and other isolates were taken over by several species of genus Bacillus. Microbiological examination of dried and polished beans resulted in the identification of 22 organisms at Centeno Estate and 15 organisms at San Louis Estate