Search results for: Bacillus licheniformis LHH100

Authors: A. Khalique, T. Naseem, N. Haque, Z. Rasool

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The objective of current study was to investigate the effect of Bacillus subtilis PB6 (CloSTAT) as a probiotic in broilers. The corn-soybean based diet was divided into four treatment groups; T1 (basal diet with no probiotic and no Clostridium perfringens); T2 (basal diet challenged with C. perfringens without probiotic); T3 (basal diet challenged with C. perfringens having 0.05% probiotic); T4 (basal diet challenged with C. perfringens having 0.1% probiotic). Every treatment group had four replicates with 24 birds each. Body weight and feed intake were measured on weekly basis, while ileal bacterial count was recorded on day-28 following Clostridium perfringens challenge. The 0.1% probiotic treatment showed 7.2% increase in average feed intake (P=0.05) and 8% increase in body weight compared to T2. In 0.1% treatment body weight was 5% higher than T3 (P=0.02). It was also observed that 0.1% treatment had improved feed conversion ratio (1.77) on 6^th week. No effect of treatment was observed on mortality and ileal bacterial count. The current study indicated that 0.1% use of probiotic had positive response in C. perfringens challenged broilers.

Keywords: Bacillus subtilis PB6, antibiotic growth promoters, Clostridium perfringens, broilers

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Authors: R. Gounina-Allouane, N. Ouali, F. Z. Berrabah, A. Bentaleb

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For a long time, the Gram-positive spore-forming bacteria Bacillus thuringiensis (Bt) has been widely used in biological control against devastating and disease vectors insects. This is due to the insecticidal activity of its crystalline parasporal inclusion (crystals) predominantly comprised of one or more proteins (Cry and Cyt proteins) also called δ-endotoxins, produced during sporulation. The shape and composition of Bt crystals vary among strains and crystalline proteins are extremely varied (more than 475 cry gene were discovered). The insecticidal activity of Bt crystals is very well studied, thus their insecticidal mode of action is well established, however, their antimicrobial effect is largely unknown. The lack of data on the antimicrobial effect of crystalline proteins of Bt and the need for searching new antimicrobial molecules encouraged us to carried out this study. The antibacterial effect of δ-endotoxines produced by two Bt stains; a strain isolated from soil at northern of Algeria (Bt 7.2.B), and a strain isolated from a bioinsecticide (Bacillus thuringiensis var aizawai), activated by proteolysis, was assayed on clinical bacterial strains and ATCC collection ones respectively. Gram positive and negative clinical bacterial strains (Escherichia coli, Klebsiella pneumonaie, Pseudomonas aeruginosa, Staphylococcus aureus) were sensitive to activated Bt 72B endotoxins. Similarly, bacterial strains from ATCC collection (Escherichia coli ATCC 25922, Pseudomonas aerugenosa ATCC 27853, Staphylococcus aureus ATCC 25923) were sensitive to activated B. thuringiensis var aizawai δ-endotoxines. The activated δ-endotoxins were separated by SDS-PAGE.

Keywords: Bacillus thuringiensis, crystals, cry proteins, δ-endotoxins, antibacterial activity

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Authors: Alok Kumar Yadav, Saurabh Saraswat, Preeti Sirohi, Manjoo Rani, Sameer Srivastava, Manish Pratap Singh, Nand K. Singh

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The development of antibiotic resistance in bacteria is increasing at an alarming rate, and this is considered as one of the most serious threats in the history of medicine, and an alternative solution should be derived so as to tackle this problem. In many countries, people use the medicinal plants for the treatment of various diseases as these are cheaper, easily available and least toxic. Syzygium cumini is used for the treatment of various kinds of diseases but their mechanism of action is not reported. The antimicrobial activity of Syzygium cumini was tested by the well diffusion assay and zone of inhibition was reported to be 20.06 mm as compared to control with MIC of 0.3 mg/ml. Genomic DNA fragmentation of Bacillus subtilis revealed apoptosis and FE-SEM indicate cell wall cracking on several intervals of time. Propidium iodide staining results showed that few bacterial cells were stained in the control and population of stained cells increase after exposing them for various period of time. Flow cytometric kinetic data analysis on the membrane permeabilization in bacterial cell showed the significant contribution of antimicrobial potential of the seed extract on antimicrobial-induced permeabilization. Two components of Syzygium cumini methanolic seed extract was found to be quite active against four enzymes like PDB ID- 1W5D, 4OX3, 3MFD and 5E2F which have a very crucial role in membrane synthesis in Bacillus subtilis by in silico analysis. Through in silico analysis, lupeol showed highest binding energy for macromolecule 1W5D and 4OX3 whereas stigmasterol showed the highest binding energy for macromolecule 3MFD and 5E2F respectively. It showed that methanolic seed extract of Syzygium cumini can be used for the inhibition of foodborne infections caused by Bacillus subtilis and also as an alternative of prevalent antibiotics.

Keywords: antibiotics, Bacillus subtilis, inhibition, Syzygium cumini

Procedia PDF Downloads 164

Authors: S. Abdulsalam, A. D. I. Suleiman, N. M. Musa, M. Yusuf

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Environment free of pollutants should be the concern of every individual but with industrialization and urbanization it is difficult to achieve. In view of achieving a pollution limited environment at low cost, a study was conducted on the use of bioremediation technology to remediate hydrocarbons and three heavy metals namely; copper (Cu), zinc (Zn) and iron (Fe) from a typical petroleum refinery wastewater in a closed system. Physicochemical and microbiological characteristics on the wastewater sample revealed that it was polluted with the aforementioned pollutants. Isolation and identification of microorganisms present in the wastewater sample revealed the presence of Bacillus subtilis, Micrococcus luteus, Staphylococcus aureus and Staphylococcus epidermidis. Bioremediation experiments carried out on five batch reactors with different compositions but at same environmental conditions revealed that treatment T5 (boosted with the association of Bacillus subtilis, Micrococcus luteus) gave the best result in terms of oil and grease content removal (i.e. 67% in 63 days). In addition, these microorganisms were able of reducing the concentrations of heavy metals in the sample. Treatments T5, T3 (boosted with Bacillus subtilis only) and T4 (boosted with Micrococcus luteus only) gave optimum percentage uptakes of 65, 75 and 25 for Cu, Zn and Fe respectively.

Keywords: boosted, bioremediation, closed system, aeration, uptake, wastewater

Procedia PDF Downloads 222

Authors: R. Gounina-Allouane, N. Ouali, F. Z. Berrabah, A. Bentaleb

Abstract:

For a long time, the Gram-positive spore-forming bacteria Bacillus thuringiensis (Bt) has been widely used in biological control against devastating and disease vectors insects. This is due to the insecticidal activity of its crystalline parasporal inclusion (crystals) predominantly comprised of one or more proteins (Cry and Cyt proteins) also called δ-endotoxins, produced during sporulation. The shape and composition of Bt crystals vary among strains and crystalline proteins are extremely varied (more than 475 cry gene were discovered). The insecticidal activity of Bt crystals is very well studied, thus their insecticidal mode of action is well established, however, their antimicrobial effect is largely unknown. The lack of data on the antimicrobial effect of crystalline proteins of Bt and the need for searching new antimicrobial molecules encouraged us to carried out this study. The antibacterial effect of δ-endotoxines produced by two Bt stains; a strain isolated from soil at northern of Algeria (Bt 7.2.B), and a strain isolated from a bioinsecticide (Bacillus thuringiensis var aizawai), activated by proteolysis, was assayed on clinical bacterial strains and ATCC collection ones respectively. Gram positive and negative clinical bacterial strains (Escherichia coli, Klebsiella pneumonaie, Pseudomonas aeruginosa, Staphylococcus aureus) were sensitive to activated Bt 72B endotoxins. Similarly, bacterial strains from ATCC collection (Escherichia coli ATCC 25922, Pseudomonas aerugenosa ATCC 27853, Staphylococcus aureus ATCC 25923) were sensitive to activated B. thuringiensis var aizawai δ-endotoxines. The activated δ-endotoxins were separated by SDS-PAGE.

Keywords: Bacillus thuringiensis, crystals, cry proteins, δ-endotoxins, antibacterial activity

Procedia PDF Downloads 409

Authors: Alok Kumar, Hari Ram, Lebin Thomas, Ved Pal Singh

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Organic solvent tolerant amylases and proteases of microbial origin are in great demand for their application in transglycosylation of water-insoluble flavanoids and in peptide synthesizing reaction in organic media. Most of the amylases and proteases are unstable in presence of organic solvent. In the present work two different bacterial strains M-11 and VP-07 were isolated from the soil sample of a dairy farm in Delhi, India, for the efficient production of extracellular amylase and protease through their screening on starch agar (SA) and skimmed milk agar (SMA) plates, respectively. Both the strains (M-11 and VP-07) were identified based on morphological, biochemical and 16S rRNA gene sequencing methods. After analysis through Ez-Taxon software, the strains M-11 and VP-07 were found to have maximum pairwise similarity of 98.63% and 100% with Bacillus subtilis subsp. inaquosorum BGSC 3A28 and Bacillus anthracis ATCC 14578 and were therefore identified as Bacillus sp. UKS1 and Bacillus sp. UKS2, respectively. Time course study of enzyme activity and bacterial growth has shown that both strains exhibited typical sigmoid growth behavior and maximum production of amylase (180 U/ml) and protease (78 U/ml) by these strains (UKS1 and UKS2) was commenced during stationary phase of growth at 24 and 20 h, respectively. Thereafter, both amylase and protease were tested for their tolerance towards organic solvents and were found to be active as well stable in p-xylene (130% and 115%), chloroform (110% and 112%), isooctane (119% and 107%), benzene (121% and 104%), n-hexane (116% and 103%) and toluene (112% and 101%, respectively). Owing to such properties, these enzymes can be exploited for their potential application in industries for organic synthesis.

Keywords: amylase, enzyme activity, industrial applications, organic solvent tolerant, protease

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Authors: Meriem Mansouri, Fatiha Bendali Saoudi, Noureddine Soltani

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Biological control presents a means of control for the protection of the environment. Bacillus thuringiensis israelensis Berliner 1915 is an inseticide of biological origin because it is a bacterium of the Bacillaceae family. This biocide has a biological importance, because of its specific larvicidal action against Culicidae, blood-sucking insects, responsible for several diseases to humans and animals through the world. As well as, its high specificity for these insects. Also, the freshwater mites, this necessarily parasitic group for aquatic species such as the Physidae, also have an effective biological control against the Culicidae, because of their voracious predation to the larvae of these insects. The present work aims to study the effects of the biocide Bacillus thuringiensis var israelinsis, against non-target adults of water mites Eylais hamata Koenike, 1897, as well as its associated host species Physa marmorata Fitzinger, 1833. After 12 days of oral treatment of adults with lethal concentration (LC50:0.08µg/ml), determined from essays on 4th instar larvae of Culex pipiens (hematophagous insects). No adverse effect has been recorded for adult individuals of Eylais hamata, contrary, snail Physa marmorata were sensitive for this dose of Bti. In parallel, after treatment at the Bti by LC50, the enzyme stress bio marker glutathione S-transferase, was measured after 24, 48 and 72 hours. The enzymatic activity of GST has increased after 24 and 48 hours following treatment.

Keywords: biological control, Bacillus thuringiensis var israelinsis, culicidae, hydrachnidia, enzymatic activity

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Authors: M. L. Ibrahim, A. Abdulhamid

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A surface active molecule synthesized by a rhizobacterial strain Bacillus lentus isolated from Cajanus cajan was investigated. The bioemulsifier was extracted, purified and partially characterized using standard methods. Surface properties of the bioemulsifier were determined by studying the emulsification index, solubility test and stability studies. Partial purification of the bioemulsifier was carried out using FT-IR analysis, Silica-gel column chromatography and thin layer chromatography. GC-MS analysis was carried out to detect the composition and mass of the lipids and esters. The isolate showed an emulsifying activity of 57% and surface activity of 36mm. The stability studies revealed that the bioemulsifier had better stability at temperature of 70oC, 8% pH and 8% NaCl concentration. FT-IR indicated the bioemulsifier to contain peptide and aliphatic chain, TLC revealed the compound to be ninhydrin positive and Column chromatography showed the presence of three amino acids namely; glutamine, valine and cysteine. GC-MS indicated the lipid moiety to contain aliphatic chain ranging from C9-C16 and two major peaks of 1,2-benzenedicarboxylic acid diethyl octyl ester. Therefore, surface active agent from Bacillus lentus can be used effectively in a wide range of applications such as in MEOR and in the biosynthesis of plasticizers for industrial uses.

Keywords: Bacillus lentus, bioemulsifiers, phthalic acid ester, Rhizosphere

Procedia PDF Downloads 387

Authors: Anza-Vhudziki Mboyi, Ilunga Kamika, Maggy Momba

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The aim of this study was to ascertain the survival limit and capability of commonly found wastewater protozoan (Aspidisca sp, Trachelophyllum sp, and Peranema sp) and bacterial (Bacillus licheniformis, Brevibacillus laterosporus, and Pseudomonas putida) species to remove COD while exposed to commercial nanomaterials under varying pH conditions. The experimental study was carried out in modified mixed liquor media adjusted to various pH levels (pH 2, 7 and 10), and a comparative study was performed to determine the difference between the cytotoxicity effects of commercial zinc oxide (nZnO) and silver (nAg) nanomaterials (NMs) on the target wastewater microbial communities using standard methods. The selected microbial communities were exposed to lethal concentrations ranging from 0.015 g/L to 40 g/L for nZnO and from 0.015 g/L to 2 g/L for nAg for a period of 5 days of incubation at 30°C (100 r/min). Compared with the absence of NMs in wastewater mixed liquor, the relevant environmental concentration ranging between 10 µg/L and 100 µg/L, for both nZnO and nAg caused no adverse effects, but the presence of 20 g of nZnO/L and 0.65 g of nAg/L significantly inhibited microbial growth. Statistical evidence showed that nAg was significantly more toxic compared to nZnO, but there was an insignificant difference in toxicity between microbial communities and pH variations. A significant decrease in the removal of COD by microbial populations was observed in the presence of NMs with a moderate correlation of r = 0.3 to r = 0.7 at all pH levels. It was evident that there was a physical interaction between commercial NMs and target wastewater microbial communities; although not quantitatively assessed, cell morphology and cell death were observed. Such phenomena suggest the high resilience of the microbial community, but it is the accumulation of NMs that will have adverse effects on the performance in terms of COD removal.

Keywords: bacteria, biological treatment, chemical oxygen demand (COD) and nanomaterials, consortium, pH, protozoan

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Authors: Samuel E. Okere, Anthony E. Ataga

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Introduction: The microbial ecology of Pleurotus osteratus and Pleurotus tuber–regium spent mushroom substrate (SMS) were characterized to determine other ways of its utilization. Materials and Methods: The microbiological properties of the spent mushroom substrate were determined using standard methods. This study was carried out at the Microbiology Laboratory University of Port Harcourt, Rivers State, Nigeria. Results: Quantitative microbiological analysis revealed that Pleurotus osteratus spent mushroom substrate (POSMS) contained 7.9x10⁵ and 1.2 x10³ cfu/g of total heterotrophic bacteria and total fungi count respectively while Pleurotus tuber-regium spent mushroom substrate (PTSMS) contained 1.38x10⁶ and 9.0 x10² cfu/g of total heterotrophic bacteria count and total fungi count respectively. The fungi species encountered from Pleurotus tuber-regium spent mushroom substrate (PTSMS) include Aspergillus and Cladosporum species, while Aspergillus and Penicillium species were encountered from Pleurotus osteratus spent mushroom substrate (POSMS). However, the bacteria species encountered from Pleurotus tuber-regium spent mushroom substrate include Bacillus, Acinetobacter, Alcaligenes, Actinobacter, and Pseudomonas species while Bacillus, Actinobacteria, Aeromonas, Lactobacillus and Aerococcus species were encountered from Pleurotus osteratus spent mushroom substrate (POSMS). Conclusion: Therefore based on the findings from this study, it can be concluded that spent mushroom substrate contain microorganisms that can be utilized both in bioremediation of oil-polluted soils as they contain important hydrocarbon utilizing microorganisms such as Penicillium, Aspergillus and Bacillus species and also as sources of plant growth-promoting rhizobacteria (PGPR) such as Pseudomonas and Bacillus species which can induce resistance on plants. However, further studies are recommended, especially to molecularly characterize these microorganisms.

Keywords: characterization, microorganisms, mushroom, spent substrate

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Authors: Mohsen Farahat, Tsuyoshi Hirajima

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Surface characteristics of Bacillus megaterium strain were investigated; zeta potential, FTIR and contact angle were measured. Surface energy components including Lifshitz-van der Waals, Hamaker constant, and acid/base components (Lewis acid/Lewis base) were calculated from the contact angle data. The results showed that the microbial cells were negatively charged over all pH regions with high values at alkaline region. A hydrophilic nature for the strain was confirmed by contact angle and free energy of adhesion between microbial cells. Adsorption affinity of the strain toward dolomite was studied at different pH values. The results showed that the cells had a high affinity to dolomite at acid pH comparing to neutral and alkaline pH. Extended DLVO theory was applied to calculate interaction energy between B. megaterium cells and dolomite particles. The adsorption results were in agreement with the results of Extended DLVO approach. Surface changes occurred on dolomite surface after the bio-treatment were monitored; contact angle decreased from 69° to 38° and the mineral’s floatability decreased from 95% to 25% after the treatment.

Keywords: Bacillus megaterium, surface modification, flotation, dolomite, adhesion energy

Procedia PDF Downloads 217

Authors: Fdhila Walid, Bayar Sihem, Khouidi Bochra, Maâtouk Fethi, Ben Amor Feten, Hajer Hentati, Mahdhi Abdelkarim

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The presence of resistant bacteria in the oral cavity can be the major cause of dental antibiotic prophylaxis failure. Multidrug efflux has been described for many organisms, including bacteria and fungi as part of their drugs resistance strategy. The potential use of probiotic bacteria can be considered as a new alternative in the prevention or cure of oral cavity diseases. In this study, different Bacillus strains isolated from the environment were isolated and characterized using biochemical and molecular procedures. The inhibitory activity against different pathogenic bacteria and yeast strains was tested using diffusion agar assay method. Our data revealed that the tested strains have an antimicrobial effect against the pathogenic strains such as Streptococcus mutants. The inhibitory effect was variable depending from the probiotic and pathogenic strains. The obtained result demonstrated that Bacillus can be used as a potential candidates probiotic and help in the prevention and treatment of oral infections, including dental caries, periodontal disease and halitosis. Our data, partly encourage the use of probiotic strains because they do not produce acid which can contribute to faster installation decay and these are spore-forming bacteria that can withstand the stress of the oral cavity (acids, alkalis, and salty foods).

Keywords: probiotic, pathogenic bacteria, yeast, oral cavity

Procedia PDF Downloads 348

Authors: G. Ultanbekova, Zh. Suleimenova, Zh. Rakhmetova, G. Mombekova, S. Mantieva

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Plant Growth Promoting Rhizobacteria (PGPR) are a group of free-living bacteria that colonize the rhizosphere, enhance plant growth of many cereals and other important agricultural crops and protect plants from disease and abiotic stresses through a wide variety of mechanisms. The use of PGPR has been proven to be an environmentally sound way of increasing crop yields by facilitating plant growth. In the present study, strain improvement of PGPR isolates were carried out by chemical mutagenesis for the improvement of growth and yield of lima bean. Induced mutagenesis is widely used for the selection of microorganisms producing biologically active substances and further improving their activities. Strain improvement is usually done by classical mutagenesis which involves exposing the microbes to chemical or physical mutagens. The strains of Pseudomonas putida 4/1, Azotobacter chroococcum Р-29 and Bacillus subtilis were subjected to mutation process for strain improvement by treatment with a chemical agent (sodium nitrite) to cause mutation and were observed for its consequent action on the seeds germination and plant growth of lima bean (Phaseolus lunatus). Bacterial mutant strains of Pseudomonas putida M-1, Azotobacter chroococcum M-1 and Bacillus subtilis M-1, treated with sodium nitrite in the concentration of 5 mg/ml for 120 min, were found effective to enhance the germination of lima bean seeds compared to parent strains. Moreover, treatment of the lima bean seeds with a mutant strain of Bacillus subtilis M-1 had a significant stimulation effect on plant growth. The length of the stems and roots of lima bean treated with Bacillus subtilis M-1 increased significantly in comparison with parent strain in 1.6 and 1.3 times, respectively.

Keywords: chemical mutagenesis, germination, kidney bean, plant growth promoting rhizobacteria (PGPR)

Procedia PDF Downloads 168

Authors: V. Khosravi, F. D. Ardejani, A. Aryafar, M. Sedighi

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Heavy metals have a damaging impact for the environment, animals and humans due to their extreme toxicity and removing them from wastewaters is a very important and interesting task in the field of water pollution control. Biosorption is a relatively new method for treatment of wastewaters and recovery of heavy metals. In this study, a continuous fixed bed study was carried out by using Bacillus thuringiensis as a biosorbent for the removal of Cu and Mn ions from Sarcheshmeh Acid Mine Drainage (AMD). The effect of operating parameters such as flow rate and bed height on the sorption characteristics of B. thuringiensis was investigated at pH 6.0 for each metal ion. The experimental results showed that the breakthrough time decreased with increasing flow rate and decreasing bed height. The data also indicated that the equilibrium uptake of both metals increased with decreasing flow rate and increasing bed height. BDST, Thomas, and Yoon–Nelson models were applied to experimental data to predict the breakthrough curves. All models were found suitable for describing the whole dynamic behavior of the column with respect to flow rate and bed height. In order to regenerate the adsorbent, an elution step was carried out with 1 M HCl and five adsorption-desorption cycles were carried out in continuous manner.

Keywords: acid mine drainage, bacillus thuringiensis, biosorption, cu and mn ions, fixed bed

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Authors: Mohammad Abdel Fattah Mohammad Kewisha

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Microbial deterioration of ancient materials became one of the biggest problems facing the workers in the field of cultural heritage protection because the microbial deterioration of artifacts causes detrimental effects on the aesthetic value of the monuments due to colonization, whether they are made of inorganic materials such as stone or organic like wood, textiles, wall paintings, and paper. So, the early identification of the bacterial strains that caused deterioration is the most important point for the protection of monument objects. The present study focuses on the Bacillus spp. group, which was isolated from some biodeterioration monuments from different areas of Egypt. The investigated objects in this study were made from organic materials (cellulose), paper, textile, and wood. Isolated strains were identified up to the species level biochemically. Eleven bacterial isolates were obtained from collected samples. They were taken from different archaeological objects, four microbicides, cetrimonium bromide, sodium azide, tetraethyl ammonium bromide, and dichloroxylenol, at various concentrations ranging from 25 ppm to 500 ppm. They were screened for their antibacterial activity against the Bacillus spp. isolates, and detection of Minimum inhibitory concentration (MIC). It was also necessary to indicate the ideal Minimum inhibitory concentration for each strain for the purpose of biotreatment of the infected monuments with less damaging effect on monument materials.

Keywords: microbial deterioration, ancient materials, heritage protection, protection of monuments, biodeteriorative monuments

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Authors: Memoona Ramzan, Bushra Tabassum, Anwar Khan, Muhammad Tariq, Mudassar Fareed Awan, Idrees Ahmad Nasir, Zahida Qamar, Naila Shahid, Tayyab Husnain

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Biological control is a novel approach being used in crop protection nowadays. Bacteria like Bacillus and Pseudomonas are reported for this purpose and few of their products are commercially available too. Rhizosphere and phyllosphere of healthy cotton plants were used as a source to isolate bacteria capable of exhibiting properties worthy for selection as biocontrol agent. For this purpose all isolated strains were screened for the activities like phosphate solubilization, Indole acetic acid (IAA) production and biocontrol against fungi. Two strains S1HL3 and S1HL4 showed phosphate solubilization and IAA production simultaneously while two other JS2HR4 and JS3HR2 were good inhibitors of fungal pathogens. Through biochemical and molecular characterization these bacteria were identified as P. aeruginosa, Burkholderia and Bacillus respectively. In green house trials of these isolates against Cotton leaf curl virus (CLCuV), seven treatments including individual bacterial isolate and consortia were included. Treated plants were healthy as compared to control plants in which upto 74% CLCuV symptomatic plants exist. Maximum inhibition of CLCuV was observed in T7 treated plants where viral load was only 0.4% as compared to control where viral load was upto 74%. This treatment consortium included Bacillus and Pseudomonas isolates; S1HL3, S1HL4, JS2HR4 and JS3HR2. Principal Component Biplot depicted highly significant correlation between percentage viral load and the disease incidence.

Keywords: cotton leaf curl virus, biological control, bacillus, pseudomonas

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Authors: N. P. Eswara Reddy, S. Thahir Basha

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Here, we report the success stories of potential leaf, seed and root endophytes against soil borne as well as foliar plant pathogens which are nutritionally adequate and safe for consumption. Endophytes are the microorganisms that reside asymptomatically in the tissues of higher plants are a robust source of potential biocontrol agents and it is presumed that the survival ability of endophytes may be better when compared to phylloplane microflora. Of all the 68 putative leaf endophytes, the endophytes viz., EB9 (100%), and EB35 (100%) which were superior in controlling Colletotrichum gloeosporioides causing mango anthracnose were identified as Brevundimonas bullata (EB09) and Bacillus thuringiensis (EB35) and further delayed in ripening of mango fruits up to 21 days. As a part, the seed endophyte GSE-4 was identified as Archoromobacter spp. against Sclerotium rolfsii causing stem rot of groundnut and the root endophyte REB-8 against Rhizoctonia bataticola causing dry root rot of chickpea was identified as Bacillus subtilis. Both recorded least percent disease incidence (PDI) and increased plant growth promotion, respectively. Further, the novel Bacillus subtilis (SEB-2) against Macrophomina pahseolina causing charcoal rot of sunflower provides an ample scope for exploring the endophytes at large scale. The talc-based formulations of these endophytes developed can be commercialized after toxicological studies. At the bottom line these unexplored endophytes are the need of the hour against aggressive plant pathogens and to maintain the quality and abundance of food and feed and also to fetch marginal economy to the farmers will be discussed.

Keywords: endophytes, plant pathogens, commercialization, abundance of food

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Authors: Ganga Sagar Bhattarai, Swastika Gurung

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Antibiotics were being misused in both humans and animals, which led to the development of multidrug-resistant microorganisms. Antibiotic abuse is likely rampant among goats, cows, and buffaloes in order to boost growth and reduce production losses. The aim of this study is to know the multidrug resistance (MDR) bacteria in goats, cows, and buffaloes. Out of 68 samples that were examined, S. aureus, Bacillus spp., E. coli, Shigella spp., Klebsiella spp., S. epidremidis, and Salmonella spp. were isolated. S. aureus was the highest isolated bacteria (91.17%), Bacillus spp. (61.76%), E. coli (48.52%), Shigella spp. (22.05%), Klebsiella spp. (17.64%), S. epidermidis (13.23%), and the Salmonella spp. (7.35%). Salmonella spp. and E. coli showed multidrug resistance to at least four antibiotics, including Amoxicillin, Tetracycline, Piperacillin, and Ciprofloxacin, in Salmonella and to at least three antibiotics, including Amoxicillin, Tetracycline, and Nalidic acid. The highest resistance bacteria Salmonella spp. showed (57.14%) E. coli and Bacillus spp. showed (42.85%) S. aureus, S. epidermidis, and Shigella spp. showed (28.57%), and Klebsiella spp. showed (14.28%). This study showed that antibiotic-resistant bacteria with high levels of Amoxicillin, Penicillin, and Tetracycline resistance are present in healthy farm animals such as goats, cows, and buffaloes. Options for antibiotic therapy in both humans and animals will likely be limited as a result. The use, distribution, storage, and sale of antibiotics in veterinary practices must consequently be under strict control.

Keywords: multidrug resistance, multidrug resistance bacteria, susceptibility testing, bacterial infections

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Authors: Khadijeh Abhari, Seyed Shahram Shekarforoush, Saeid Hosseinzadeh

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An in vivo trial was conducted to evaluate the effects of Bacillus coagulans, and inulin, either separately or in combination, on lipid profile using a rat model. Thirty-two male Wistar rats were randomly divided into four groups (n=8) and fed as follows: standard diet (control), standard diet with 5% w/w long chain inulin (prebiotic), standard diet with 109 spores/day spores of B. coagulans by orogastric gavage (probiotic), and standard diet with 5% w/w long chain inulin and 109 spores/day of B. coagulans (synbiotic). Rats were fed the treatments for 30 days. Serum samples were collected 10, 20 and 30 days following onset of treatment. Total cholesterol, HDL and LDL cholesterol and triglycerides concentrations were analyzed. Results of this study showed that inulin potentially affected the lipid profile. An obvious decrease in serum total cholesterol and LDL-cholestrol of rats fed with inulin in synbiotic and prebiotic groups was seen in all sampling days. Inulin fed rats also demonstrated higher levels of HDL-cholesterol concentration; however this value in probiotic and control fed rats remains without significant change. According to the results of this study, B. coagulans did not contribute to any lipid profile changes after 30 days. Thus, further in vitro investigations on the characteristic of these bacteria could be useful to gain insights into understanding the treatment of probiotics in order to achieve the maximum beneficial effect.

Keywords: bacillus coagulans, inulin, rat, lipid profile, synbiotic diet

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Authors: Aida Kalantari, Boyang Ji, Tao Chen, Ivan Mijakovic

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3-hydroxypropanoic acid (3-HP) is one of the most important biomass-derivable platform chemicals that can be converted into a number of industrially important compounds. There have been several attempts at production of 3-HP from renewable sources in cell factories, focusing mainly on Escherichia coli, Klebsiella pneumoniae, and Saccharomyces cerevisiae. Despite the significant progress made in this field, commercially exploitable large-scale production of 3-HP in microbial strains has still not been achieved. In this study, we investigated the potential of Bacillus subtilis to be used as a microbial platform for bioconversion of glycerol into 3-HP. Our recombinant B. subtilis strains overexpress the two-step heterologous pathway containing glycerol dehydratase and aldehyde dehydrogenase from various backgrounds. The recombinant strains harboring the codon-optimized synthetic pathway from K. pneumoniae produced low levels of 3-HP. Since the enzymes in the heterologous pathway are sensitive to oxygen, we had to perform our experiments in micro-aerobic conditions. Under these conditions, the cell produces lactate in order to regenerate NAD+, and we found the lactate production to be in competition with the production of 3-HP. Therefore, based on the in silico predictions, we knocked out the glycerol kinase (glpk), which in combination with growth on glucose, resulted in improving the 3-HP titer to 1 g/L and the removal of lactate. Cultivation of the same strain in an enriched medium improved the 3-HP titer up to 7.6 g/L. Our findings provide the first report of successful introduction of the biosynthetic pathway for conversion of glycerol into 3-HP in B. subtilis.

Keywords: bacillus subtilis, glycerol, 3-hydroxypropanoic acid, metabolic engineering

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Authors: Yan-Shiang Chiou, Yi-Huang Hsueh

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Bacillus cereus is a foodborne pathogen that causes two types of foodborne illness, the emetic and diarrheal syndromes. B. cereus consistently ranks among the top three among bacterial foodborne outbreaks in the ten years of 2001 to 2010 in Taiwan. Foodborne outbreak caused by B. cereus has been increased, and recently it ranks second foodborne pathogen after Vibrio parahaemolyticus. This pathogen is difficult to control due to its ubiquitousness in the environment, the psychrotrophic nature of many strains, and the heat resistance of their spores. Because complete elimination of biofilms is difficult, a better understanding of the molecular mechanisms of biofilm formation by B. cereus will help to develop better strategies to control this pathogen. Surface translocation can be an important factor in biofilm formation. In B. cereus, NprR is a quorum sensor, and its apo NprR is a dimer and changes to a tetramer in the presence of NprX. The small peptide NprX may induce conformational change allowing the apo dimer to switch to an active tetramer specifically recognizing target DNA sequences. Our result showed that mutation of nprRX causes surface spreading deficiency. Mutation of flagella, pili and surfactant genes (flgAB, bcpAB, krsABC), did not abolish spreading motility. Under nprRX mutant, mutation of spo0A restored the spreading deficiency. This suggests that spreading motility is not related surfactant, pili and flagella but other unknown mechanism and Spo0A, a sporulation initiation protein, inhibits spreading motility.

Keywords: Bacillus cereus, nprRX, spo0A, spreading motility

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Authors: Nice Vasconcelos Coimbra, Fábio dos Santos Gonçalves, Marisa Nascimento, Ellen Cristine Giese

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The development of chemical routes for the recovery and separation of rare earth elements (REE) is seen as a priority and strategic action by several countries demanding these elements. Among the possibilities of alternative routes, the biosorption process has been evaluated in our laboratory. In this theme, the present work attempts to assess and fit the solution equilibrium data in Langmuir, Freundlich and DKR isothermal models, based on the biosorption results of the lanthanum and samarium elements by Bacillus subtilis immobilized on calcium alginate gel. It was observed that the preference of adsorption of REE by the immobilized biomass followed the order Sm (III)> La (III). It can be concluded that among the studied isotherms models, the Langmuir model presented better mathematical results than the Freundlich and DKR models.

Keywords: rare earth elements, biosorption, Bacillus subtilis, adsorption isotherm models

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Authors: Dong Hwi Kim, Dharaneedharan Subramanian, So Hyun Park, Ha-Ri Choi, Ji-Hyung Kim, Dong-Hoon Lee, Moon Soo Heo

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The present study was performed to evaluate the use of Bacillus amyloliquefaciens JFP-2 isolated from a traditional fermented sea food, as probiotic bacteria in the diets for Rock-bream, Oplegnathus faciatus. A total of 180 fish (187.4 ± 2.7 g) were divided into two groups, control (C) and probiotic (P) group (90 fish per group) in triplicate. C group was fed with basal diet without probiotic, while P group was fed with B. amyloliquefaciens spores at concentration of 1.4 x 106 colony forming units per gram (CFU/g) of feed. After two months of feeding experiments, P group fish showed significant improvements in body weight (BW), weight gain (WG), specific growth rate (SGR) and food conversion ratio (FCR) compared with C group. Also, bi-weekly assessment of serum protein, glucose, fatty acid profile showed a significant increase in probiotic fed fish than that of control fish group. Similar increase in serum antioxidant and lysozyme activity was found in probiotic fed fish group. Twenty days challenge experiment shows decrease mortality in probiotic fed fish group when compared with that of control group. Hence, these results indicate that the use of B. amyloliquefaciens JFP-2 as a feed supplement, is beneficial to improve the health status of Oplegnathus fasciatus challenged with Vibrio anguillarum.

Keywords: Bacillus amyloliquefaciens, Oplegnathus fasciatus, probiotic feed, rock bream

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Authors: Tahammal Hussain, Khuram Zia, Mumammad Jalal Arif, Megha Parajulee, Abdul Hakeem

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The Bioassay on neonate, 2nd and 3rd instar larvae of Helicoverpa armigera (Hubner) were conducted against Bacillus thuringiensis proteins Cry1Ac. Cry1Ac was incorporated into an artificial diet and was serially diluted with distilled water and then mixed with diet at an appropriate temperature of diet. Toxins incorporated prepared diet was poured into Petri-dishes. For controls, distilled water was mixed with the diet. Five toxin doses 0.25, 0.5, 1, 2, and 4 ug / ml and one control were used for each instars of H. armigera 20 larvae were used in each replication and each treatment is replicated four times. LC50 of Cry1Ac against neonate, 2nd and 3rd instar larvae of H. armigera were 0.34, 0.81 and 1.46 ug / ml. So Cry1Ac is more effective against neonate larvae of H .armigera as compared to 2nd and 3rd instar larvae under laboratory conditions.

Keywords: B. thuringiensis, Cry1Ac, H. armigera, toxicity

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Authors: Sneha Singh, Abhimanyu Dev, Vinod Nigam

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The major issue which decides the impending use of gold nanoparticles (AuNPs) in nanobiotechnological applications is their particle size and stability. Often the AuNPs obtained biomimetically are considered useless owing to their instability in the aqueous medium and thereby limiting the widespread acceptance of this facile green synthesis procedure. So, the use of nontoxic surfactants is warranted to stabilize the biogenic nanoparticles (NPs). But does the surfactant only play a role in stabilizing by being adsorbed to the NPs surface or can it have any other significant contribution in synthesis process and controlling their size as well as shape? Keeping this idea in mind, AuNPs were synthesized by using surfactant treated (lechate) and untreated (cell lysate supernatant) Bacillus licheniformis cell extract. The cell extracts mediated reduction of chloroauric acid (HAuCl 4) in the presence of non-ionic surfactant, Tween 20 (TW20), and its effect on the AuNPs stability was studied. Interestingly, the surfactant used in the study served as potential alternative to harvest cellular enzymes involved in bioreduction process in a hassle free condition. The surfactants ability to solubilize/leach membrane proteins and simultaneously stabilizing the AuNPs could have advantage from process point of view as it will reduce the time and economics involve in the nanofabrication of biogenic NPs. The synthesis was substantiated with UV-Vis spectroscopy, Dynamic light scattering study, FTIR spectroscopy, and Transmission electron microscopy. The Zeta potential of AuNPs solutions was measured routinely to corroborate the stability observations recorded visually. Highly stable, ultra-small AuNPs of 2.6 nm size were obtained from the study. Further, the biological efficacy of the obtained AuNPs as potential antibacterial agent was evaluated against Bacilllus subtilis, Pseudomonas aeruginosa, and Escherichia coli by observing the zone of inhibition. This potential of AuNPs of size < 3 nm as antibacterial agent could pave way for development of new antimicrobials and overcoming the problems of antibiotics resistance

Keywords: antibacterial, bioreduction, nanoparticles, surfactant

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Authors: N. Kotabin, Y. Tahara, K. Issakul, O. Chunhachart

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Cadmium (II) (Cd) is one of the major toxic elemental pollutants which is hazardous for humans, animals and plants. γ-Polyglutamic acid (γ-PGA) is an extracellular biopolymer produced by several species of Bacillus which has been reported to be an effective biosorbent for metal ions. The effect of γ-PGA on growth of rice grown under laboratory conditions was investigated. Rice seeds were germinated and then grown at 30±1°C on filter paper soaked with Cd solution and γ-PGA for 7 days. The result showed that Cd significantly inhibited the growth of roots and shoots by reducing root and shoot lengths. Fresh and dry weights also decreased compared with control; however, the addition of 500 mg•L-1 γ-PGA alleviated rice seedlings from the adverse effects of Cd. The analysis of physiological traits revealed that Cd caused a decrease in the total chlorophyll and soluble protein contents and amylase activities in all treatments. The Cd content in seedling tissues increased for the Cd 250 μM treatment (P < 0.05) but the addition of 500 mg•L-1 γ-PGA resulted in a noticeable decrease in Cd (P < 0.05).

Keywords: polyglutamic acid, cadmium, rice, bacillus subtilis

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Authors: Nonhlanhla Nkosi, Kulsum Kondiah

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The toxicological manifestation of heavy metals motivates interest towards the development of a reliable, eco-friendly biosorption process. With that being said, the aim of the current study was to characterise the EPS from heavy-metal resistant bacteria isolated from acid mine decant on the West Rand, Gauteng, South Africa. To achieve this, six exopolysaccharide (EPS) producing, metal resistant strains (Pb101, Pb102, Pb103, Pb204, Co101, and Ni101) were identified as Bacillus safensis strain NBRC 100820, Bacillus proteolyticus, Micrococcus luteus, Enterobacter sp. Pb204, Bacillus wiedmannii and Bacillus zhangzhouensis, respectively with 16S rRNA sequencing. Thereafter, EPS was extracted using chemical (formaldehyde/NaOH) and physical (ultrasonification) methods followed by physicochemical characterisation of carbohydrate, DNA, and protein contents using chemical assays and spectroscopy (FTIR- Fourier transformed infrared and 3DEEM- three-dimensional excitation-emission matrix fluorescence spectroscopy). EPS treated with formaldehyde/NaOH showed better recovery of macromolecules than ultrasonification. The results of the present study showed that carbohydrates were more abundant than proteins, with carbohydrate and protein concentrations of 8.00 mg/ml and 0.22 mg/ml using chemical method in contrast to 5.00 mg/ml and 0.77 mg/ml using physical method, respectively. The FTIR spectroscopy results revealed that the extracted EPS contained hydroxyl, amide, acyl, and carboxyl groups that corresponded to the aforementioned chemical analysis results, thus asserting the presence of carbohydrates, DNA, polysaccharides, and proteins in the EPS. These findings suggest that identified functional groups of EPS form surface charges, which serve as the binding sites for suspended particles, thus possibly mediating adsorption of divalent cations and heavy metals. Using the extracted EPS in the development of a cost-effective biosorption solution for industrial wastewater treatment is attainable.

Keywords: biosorbent, exopolysaccharides, heavy metals, wastewater treatment

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Authors: Togzhan D. Mukasheva, Anel A. Omirbekova, Raikhan S. Sydykbekova, Ramza Zh. Berzhanova, Lyudmila V. Ignatova

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Following plants-barley (Hordeum sativum), alfalfa (Medicago sativa), grass mixture (red fescue-75%, long-term ryegrass - 20% Kentucky bluegrass - 10%), oilseed rape (Brassica napus biennis), resistant to growth in the contaminated soil with oil content of 15.8 g / kg 25.9 g / kg soil were used. Analysis of the population showed that the oil pollution reduces the number of bacteria in the rhizosphere and rhizoplane of plants and enhances the amount of spore-forming bacteria and saprotrophic micromycetes. It was shown that regardless of the plant, dominance of Pseudomonas and Bacillus genera bacteria was typical for the rhizosphere and rhizoplane of plants. The frequency of bacteria of these genera was more than 60%. Oil pollution changes the ratio of occurrence of various types of bacteria in the rhizosphere and rhizoplane of plants. Besides the Pseudomonas and Bacillus genera, in the presence of hydrocarbons in the root zone of plants dominant and most typical were the representatives of the Mycobacterium and Rhodococcus genera. Together the number was between 62% to 72%.

Keywords: pollution, root system, micromycetes, identification

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Authors: Kgaugelo Lekota, Ayesha Hassim, Henriette Van Heerden

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Bacillus anthracis is the causative agent of anthrax that is composed of three genetic groups, namely A, B, and C. Clade-A is distributed world-wide, while sub-clades B has been identified in Kruger National Park (KNP), South Africa. KNP is one of the endemic anthrax regions in South Africa with distinctive genetic diversity. Genomic surveillance of KNP B. anthracis strains was employed on the historical culture collection isolates (n=67) dated from the 1990’s to 2015 using a whole genome sequencing approach. Whole genome single nucleotide polymorphism (SNPs) and pan-genomics analysis were used to define the B. anthracis genetic population structure. This study showed that KNP has heterologous B. anthracis strains grouping in the A-clade with more prominent ABr.005/006 (Ancient A) SNP lineage. The 2012 and 2015 anthrax isolates are dispersed amongst minor sub-clades that prevail in non-stabilized genetic evolution strains. This was augmented with non-parsimony informative SNPs of the B. anthracis strains across minor sub-clades of the Ancient A clade. Pan-genomics of B. anthracis showed a clear distinction between A and B-clade genomes with 11 374 predicted clusters of protein coding genes. Unique accessory genes of B-clade genomes that included biosynthetic cell wall genes and multidrug resistant of Fosfomycin. South Africa consists of diverse B. anthracis strains with unique defined SNPs. The sequenced B. anthracis strains in this study will serve as a means to further trace the dissemination of B. anthracis outbreaks globally and especially in South Africa.

Keywords: bacillus anthracis, whole genome single nucleotide polymorphisms, pangenomics, kruger national park

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Authors: Arfan Ali, Idrees Ahmad Nasir

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The critical evaluation of tolerance in tomato plants against the induced saline soil were assessed by transcript analysis of genes coding for products potentially involved in stress tolerance. A reverse transcriptase PCR experiment was performed with Hsp90-1, MT2, and GR1like protein genes using RNA isolated from different tissues of tomato plants. Four strains of Bacillus magisterium were inoculated with 100 Mm & 200 Mm concentrations of salt. Eleven treatments each ten replica pots were installed in green house experiment and the parameters taken into account were morphological (length, weight, number of leaves, leaf surface area), chemical (anthocyanin, chlorophyll-a, chlorophyll-b, carotenoids) and biological (gene expression). Results bare a response i.e. highest response of MT2 like gene was at 24 hpi and the highest levels of GR1 like protein transcript accumulation were detected at 36 hpi. The chemical and morphological parameters at diverse salt concentrations bequeath superlative response amongst strains which candidly flank on Zm7 and Zm4. Therefore, Bacillus magisterium Zm7 strains and somehow Zm4 strain can be used in saline condition to make plants tolerant. The overall performance of strains Zm7, Zm6, and Zm4 was found better for all studied traits under salt stress conditions. Significant correlations among traits root length, shoot length, number of leaves, leaf surface area, carotenoids, anthocyanin, chlorophyll-a and chlorophyll-b were found and suggested that the salt tolerance in tomato may be improved through the use of PGPR strains.

Keywords: Bacillus magisterium, gene expression glutathione reductase, metallothionein, PGPR, Rhizobacteria, saline

Procedia PDF Downloads 406