Search results for: plant related to bacillus bacteria.

Authors: Assel E. Molzhigitova, Amankeldi K. Sadanov, Elvira T. Ismailova, Kulyash A. Iskandarova, Olga N. Shemshura, Ainur I. Seitbattalova

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Fire blight is a very harmful for commercial apple and pear production quarantine bacterial disease. To date, several different methods have been proposed for disease control, including the use of copperbased preparations and antibiotics, which are not always reliable or effective. The use of bacteria as biocontrol agents is one of the most promising and eco-friendly alternative methods. Bacteria with protective activity against the causal agent of fire blight are often present among the epiphytic microorganisms of the phyllosphere of host plants. Therefore, the main objective of our study was screening of local epiphytic bacteria as possible antagonists against Erwinia amylovora, the causal agent of fire blight. Samples of infected organs of apple and pear trees (shoots, leaves, fruits) were collected from the industrial horticulture areas in various agro-ecological zones of Kazakhstan. Epiphytic microorganisms were isolated by standard and modified methods on specific nutrient media. The primary screening of selected microorganisms under laboratory conditions to determine the ability to suppress the growth of Erwinia amylovora was performed by agar-diffusion-test. Among 142 bacteria isolated from the fire blight host plants, 5 isolates, belonging to the genera Bacillus, Lactobacillus, Pseudomonas, Paenibacillus and Pantoea showed higher antagonistic activity against the pathogen. The diameters of inhibition zone have been depended on the species and ranged from 10 mm to 48 mm. The maximum diameter of inhibition zone (48 mm) was exhibited by B. amyloliquefaciens. Less inhibitory effect was showed by Pantoea agglomerans PA1 (19 mm). The study of inhibitory effect of Lactobacillus species against E. amylovora showed that among 7 isolates tested only one (Lactobacillus plantarum 17M) demonstrated inhibitory zone (30 mm). In summary, this study was devoted to detect the beneficial epiphytic bacteria from plants organs of pear and apple trees due to fire blight control in Kazakhstan. Results obtained from the in vitro experiments showed that the most efficient bacterial isolates are Lactobacillus plantarum 17M, Bacillus amyloliquefaciens MB40, and Pantoea agglomerans PA1. These antagonists are suitable for development as biocontrol agents for fire blight control. Their efficacies will be evaluated additionally, in biological tests under in vitro and field conditions during our further study.

Keywords: antagonists, epiphytic bacteria, Erwinia amylovora, fire blight

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Authors: Falana Yetunde Olaitan, Ijah U. J. J, Solebo Shakirat O.

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Genetically modified crops are already phenomenally successful and are grown worldwide in more than eighteen countries on more than 67 million hectares. Nigeria, in October 2018, approved Bacillus thuringiensis (Bt) cotton and maize; therefore, the need to carry out environmental risk assessment studies. A total of 15 4L octagonal ceramic pots were filled with 4kg of soil and placed on the bench in 2 rows of 10 pots each and the 3rd row of 5 pots, 1st-row pots were used to plant GM cotton seeds, while the 2nd-row pots were used for non-GM cotton seeds and the 3rd row of 5 pots served as control, all in the screen house. Soil samples for metagenomic DNA extraction were collected at random and at the monthly interval after planting at a distance of 2mm from the plant’s root and at a depth of 10cm using a sterile spatula. Soil samples for physicochemical analysis were collected before planting and after harvesting the GM and non-GM crops as well as from the control soil. The DNA was extracted, quantified and sequenced; Sample 1A (DNA from GM cotton Soil at 1st interval) gave the lowest sequence read with 0.853M while sample 2B (DNA from GM cotton Soil at 2nd interval) gave the highest with 5.785M, others gave between 1.8M and 4.7M. The samples treatment were grouped into four, Group 1 (GM cotton soil from 1 to 3 intervals) had between 800,000 and 5,700,000 strains of microbes (SOM), Group 2 (non GM cotton soil from 1 to 3 intervals) had between 1,400,600 and 4,200,000 SOM, Group 3 (control soil) had between 900,000 and 3,600,000 SOM and Group 4 (initial soil) had between 3,700,000 and 4,000,000 SOM. The microbes observed were predominantly bacteria (including archaea), fungi, dark matter alongside protists and phages. The predominant bacterial groups were the Terrabacteria (Bacillus funiculus, Bacillus sp.), the Proteobacteria (Microvirga massiliensis, sphingomonas sp.) and the Archaea (Nitrososphaera sp.), while the fungi were Aspergillus fischeri and Fusarium falciforme. The comparative analysis between groups was done using JACCARD PERMANOVA beta diversity analysis at P-value not more than 0.76 and there was no significant pair found. The pH for initial, GM cotton, non-GM cotton and control soil were 6.28, 6.26, 7.25, 8.26 and the percentage moisture was 0.63, 0.78, 0.89 and 0.82, respectively, while the percentage Nitrogen was observed to be 17.79, 1.14, 1.10 and 0.56 respectively. Other parameters include, varying concentrations of Potassium (0.46, 1,284.47, 1,785.48, 1,252.83 mg/kg) and Phosphorus (18.76, 17.76, 16.87, 15.23 mg/kg) were recorded for the four treatments respectively. The soil consisted mainly of silt (32.09 to 34.66%) and clay (58.89 to 60.23%), reflecting the soil texture as silty – clay. The results were then tested with ANOVA at less than 0.05 P-value and no pair was found to be significant as well. The results suggest that the GM crops have no significant effect on microbial ecology and physicochemical properties of the soil and, in turn, no direct or indirect effects on human health.

Keywords: genetically modified crop, microbial ecology, physicochemical properties, metagenomics, DNA, soil

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Authors: Yaara Oppenheimer-Shaanan, Nimrod Nachmias, Marina Campos Rocha, Neta Schlezinger, Noam Dotan, Asaf Levy

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Fusarium oxysporum, a fungus that attacks a broad range of plants and can cause infections in humans, operates across different kingdoms. This pathogen encounters varied conditions, such as temperature, pH, and nutrient availability, in plant and human hosts. The Fusarium oxysporum species complex, pervasive in soils globally, can affect numerous plants, including key crops like tomatoes and bananas. Controlling Fusarium infections can involve biocontrol agents that hinder the growth of harmful strains. Our research developed a computational method to identify toxin domains within a vast number of microbial genomes, leading to the discovery of nine distinct toxins capable of killing bacteria and fungi, including Fusarium. These toxins appear to function as enzymes, causing significant damage to cellular structures, membranes and DNA. We explored biological control using bacteria that produce polymorphic toxins, finding that certain bacteria, non-pathogenic to plants, offer a safe biological alternative for Fusarium management, as they did not harm macrophage cells or C. elegans. Additionally, we elucidated the 3D structures of two toxins with their protective immunity proteins, revealing their function as unique DNases. These potent toxins are likely instrumental in microbial competition within plant ecosystems and could serve as biocontrol agents to mitigate Fusarium wilt and related diseases.

Keywords: microbial toxins, antifungal, Fusarium oxysporum, bacterial-fungal intreactions

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Authors: Adamu Muhammed Tukur

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Pulp and paper industry is one of the fastest growing industries due to an increased demand in paper products. For it to satisfy this ever increasing demand, it adopts new technological innovations some of which are proved to affect our environment negatively. Global consumption of paper has increased by 400% in the last four decades and this suggests that more research is required to assess the impact of industrial effluents to our environment and public health. Paper products are generally biodegradable, however, the processes involved in its production which involve the use of mainly bleaching agents and other non-biodegradable substances pose serious problem to the environment. There are more than 250 chemicals released in paper mill waste and some are xenobiotics. Different methods such as physical and chemical methods can be adopted for the remediation of the effluents but are proved to be costly and not safe to the environment. On the other hand, biological method is shown to be less costly and environmentally friendly. Microorganisms and their enzymes have shown a promising future for bioremediation of effluents related to paper mill. Many studies prove that one of the major pollutants in the paper mill effluent is phenol especially its chlorinated derivatives. Pentachlorophenol is extremely hazardous to living cells and therefore need to be removed from the environment. Microorganisms including bacteria and fungi have the potential to degrade phenolic compounds e.g. Bacillus stearothermiphilus, Pseudomonas putida, Coricus versicolor, Sphingomonas chlorophenolica, Fusarium sp, Bacillus subtilis and P. aeroginosa. Enzymes used for the degradation include phenol hydrooxylase, polyphenoloxylase, laccase, peroxidase among others. Lignin is another important pollutant and is resistant to microbial degradation but it has been proved that certain bacteria and fungi like can degrade it. Among the fungi white-rot fungi like Fomes lividus and Trametes vesicolor are the most important bioremediators. This review focused on use of microorganism to reduce or eradicate pollutants released from the paper industry. It can serve as a review for further research to be conducted especially in the field of Biotechnology.

Keywords: bioremediation, pulp and paper, pentachlorophenol, environment

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Authors: Ali M. Zaid, Muftah Mayouf, Yahya Said Farouj

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The effect of post emergence herbicides on soil micro-flora and nitrogen fixing bacteria was studied in pea field. Pea (Pisum sativum) was grown and treated with one or a mixture of two of several herbicides 2 weeks after sowing. Soil samples were collected 2 weeks after herbicides application. Average number of colony forming units per gram of soil of bacteria, actinomycetes and fungi were determined. Average number of nodules per plant was obtained at the end of the growing season. The results of the study showed MCPB, Bentazon, MCPB+Fluozifop-p-butyl, Bentazon+Fluozifop-p-butyl, Metribuzin, Flouzifop-p-butyl+Metribuzin, Cycloxydin, and Sethoxydin increased the population of soil fungi, with 4 to 10 times compared with the control. The herbicides used showed no significant effects on nitrogen fixing bacteria. The effects of herbicides on soil bacteria and actinomycetes were different. The study showed the use of herbicides could influence the biological balance of soil microflora, which has an important role in soil fertility and microbial ecosystem.

Keywords: herbicides, post emergence, nitrogen fixing bacteria, environmental systems

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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: 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

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Authors: Mohamed Trigui, Fatma Masmoudi, Imen Zouari

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Massive utilizations of chemical fertilizer and chemical pesticides in agriculture sector to improve the farming productivity have created increasing environmental damages. Then, agriculture must become sustainable, focusing on production systems that respect the environment and help to reduce climate change. Isolation and microbial identification of new bacterial strains from naturally saline habitats and compost extracts could be a prominent way in pest management and crop production under saline conditions. In this study, potential mechanisms involved in plant growth promotion and suppressive activity against fungal diseases of a compost extract produced from poultry manure/olive husk compost and halotolerant and halophilic bacterial strains under saline stress were investigated. On the basis of the antimicrobial tests, different strains isolated from Sfax solar saltern (Tunisia) and from compost extracts were selected and tested for their plant growth promoting traits, such as siderophores production, nitrogen fixation, phosphate solubilization and the production of extracellular hydrolytic enzymes (protease and lipase) under in-vitro conditions. Among 450 isolated bacterial strains, 16 isolates showed potent antifungal activity against the tested plant pathogenic fungi. Their identification based on 16S rRNA gene sequence revealed they belonged to different species. Some of these strains were also characterized for their plant growth promoting capacities. Obtained results showed the ability of four strains belonging to Bacillus genesis to ameliorate germination rate and root elongation compared to the untreated positive controls. Combinatorial capacity of halotolerant bacteria with antimicrobial activity and plant growth promoting traits could be promising sources of interesting bioactive substances under saline stress.

Keywords: abiotic stress, biofertilizer, biotic stress, compost extract, halobacteria, plant growth promoting (PGP), soil fertility

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Authors: B. Samuel Raj, Solomon R. D. Jebakumar

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Microbial fuel cells (MFCs) are an emerging and promising method for achieving sustainable energy since they can remove contaminated organic matter and simultaneously generate electricity. Our approach was driven in three different ways like Bacterial fuel cell, Soil Microbial fuel cell (Soil MFC) and Plant Microbial fuel cell (Plant MFC). Bacterial MFC: Sulphate reducing bacteria (SRB) were isolated and identified as the efficient electricigens which is able to produce ±2.5V (689mW/m2) and it has sustainable activity for 120 days. Experimental data with different MFC revealed that high electricity production harvested continuously for 90 days 1.45V (381mW/m2), 1.98V (456mW/m2) respectively. Biofilm formation was confirmed on the surface of the anode by high content screening (HCS) and scanning electron Microscopic analysis (SEM). Soil MFC: Soil MFC was constructed with low cost and standard Mudwatt soil MFC was purchased from keegotech (USA). Vermicompost soil (V1) produce high energy (± 3.5V for ± 400 days) compared to Agricultural soil (A1) (± 2V for ± 150 days). Biofilm formation was confirmed by HCS and SEM analysis. This finding provides a method for extracting energy from organic matter, but also suggests a strategy for promoting the bioremediation of organic contaminants in subsurface environments. Our Soil MFC were able to run successfully a 3.5V fan and three LED continuously for 150 days. Plant MFC: Amaranthus candatus (P1) and Triticum aestivium (P2) were used in Plant MFC to confirm the electricity production from plant associated microbes, four uniform size of Plant MFC were constructed and checked for energy production. P2 produce high energy (± 3.2V for 40 days) with harvesting interval of two times and P1 produces moderate energy without harvesting interval (±1.5V for 24 days). P2 is able run 3.5V fan continuously for 10days whereas P1 needs optimization of growth conditions to produce high energy.

Keywords: microbial fuel cell, biofilm, soil microbial fuel cell, plant microbial fuel cell

Procedia PDF Downloads 312

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: L. C. Sánchez, L. C. Corrales, A. G. Lancheros, E. Castañeda, Y. Ariza, L. S. Fuentes, L. Sierra, J. L. Cuervo

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The importance of environment friendly alternatives in agricultural processes is the reason why the research group Ceparium, the Colegio Mayor de Cundinamarca University, Colombia, investigated the genus Bacillus and its applicability for improving crops of economic importance in Colombia. In this investigation, we presented a study in which the genus Bacillus plays a leading role as beneficial microorganism. The objective was to identify the biochemical potential of three indigenous species of Bacillus, which were able to carry out actions for biological control against pathogens and pests or promoted growth to improve productivity of crops in Colombia. The procedures were performed in three phases: first, the production of biomass of an indigenous strain and a reference strain starting from culture media for production of spores and toxins were made. Spore count was done in a Neubauer chamber, concentrations of spores of Bacillus sphaericus were prepared and a bioassay was done at the Laboratory of Entomology at the University Jorge Tadeo Lozano of Plutella xylostella larvae, insect pest of crucifers in several Colombian regions. The second phase included the extraction in the liquid state fermentation, a secondary metabolite that has antibiosis action against fungi, call iturin B, and was obtained from strains of Bacillus subtilis. The molecule was identified using High Resolution Chromatography (HPLC) and its biocontrol effect on Fusarium sp fungus causes vascular wilt in economically important plant varieties, was confirmed using testing of antagonism in Petri dish. In the third phase, an initial procedure in that let recover and identify microorganisms of the genus Bacillus from the rhizosphere in two aromatic herbs, Rosmarinus officinalis and Thymus vulgaris L. was used. Subsequently, testing of antagonism against Fusarium sp were made and an assay was done under greenhouse conditions to observe biocontrol and growth promoting action by comparing growth in length and dry weight. In the first experiment, native Bacillus sphaericus was lethal to 92% Plutella xylostella larvae in 10 DDA. In the second experiment, iturin B was identified and biological control of Fusarium sp was demonstrated. In the third study, all strains demonstrated biological control and the B14 strain identified as Bacillus megaterium increased root length and productivity of the two plants in terms of weight. It was concluded that the native microorganisms of the genus Bacillus has a great biochemical potential that provides a beneficial interactions with plants, improve their growth and development and therefore a greater impact on production.

Keywords: genus bacillus, biological control, PGPRs, biochemical potential

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Authors: Muhammad Shahzad Tufail, Iram Liaqat, Umer Sohail Meer, Muhammad Ishtaiq, Muhammad Sattar

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Nanotechnology is a vibrant field with numerous applications in many different branches of science and technology. Several methods are used to synthesize nanoparticles (NPs), which have multiple range of applications. Comparatively, the biogenic synthesis of NPs is a more economical and environmentally favourable method than the traditional chemical method. The current study aims to synthesize biogenically silver nanoparticles (AgNPs) using bacterial isolates. Four bacterial strains Escherichia coli (MT448673), Pseudomonas aeruginosa (MN900691), Bacillus subtilis (MN900684) and Bacillus licheniformis (MN900686) were used for the synthesis of AgNPs from silver nitrate (AgNO3) solution. The biofilm time kinetics of four bacterial isolates (P. aeruginosa, E. coli, B. licheniformis and B. subtilis) was analysed by incubating bacterial cultures at 37◦C in test tubes over a period of different time intervals i.e., 2, 3, 5 and 7 days following crystal violet staining method. All the four strains had ability to form strong biofilms between 48 to 72 hours of incubation. Two strains (B. subtilis and B. licheniformis) formed significant (p < 0.05) biofilm after 3 days of incubation period. The other two strains (E. coli and P. aeruginosa) showed strong biofilm formation after 2 days of incubation. Next, the antibiofilm activity of biogenically synthesized AgNPs (10 - 100 µgmL-1) was analysed against biofilm forming human pathogenic bacteria. Findings of the work revealed that 60-90% inhibition was observed at 60 µgmL-1 of AgNPs, while maximum inhibition (i.e.,100%) was found at highest concentration (90 µgmL-1). It was evident that highly significant (p < 0.05) decrease in biofilm formation was observed with increasing concentration of AgNPs.

Keywords: antibiofilm, biofilm formation, nanotechnology, pathogenic bacteria, silver nanoparticles

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Authors: Tanju Teker, Yener Tekeli̇, Esra Karpuz

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Salvia species are known as medicinal plant and often used in public. The antimicrobial effects and essential oil composition of Salvia verticillata L. subsp. amasiaca were determined. The antimicrobial activity is determined by using disk diffusion method against two Gram-positive bacteria, two Gram-negative bacteria and one kind of yeast and essential oil composition was determined by GC - MS. As a result of antimicrobial analysis while sample has shown very strong antimicrobial activity against Staphylococcus aureus, moderately effective against Pseudomonas aeruginosa and low effective against Enterococcus faecalis, it has not shown antimicrobial activity against Escherichia coli and C. albicans. Trans-caryophyllene (% 35.07), germacrene-d (% 10.98) and caryopyllene oxide (% 5.81) are the main components of essential oil composition.

Keywords: salvia, medicinal plant, antimicrobial activity, essential oil

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Authors: S. Hannah Elizabeth, A. Panneerselvam

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Objective: The main objective was to evaluate the degradative properties of Bacillus cereus sp TUHP2 isolated from TNT-Polluted soils in the Vellore District, Tamil Nadu, India. Methods: Among the 3 bacterial genera isolated from different soil samples, one potent TNT degrading strain Bacillus cereus sp TUHP2 was identified. The morphological, physiological and the biochemical properties of the strain Bacillus cereus sp TUHP2 was confirmed by conventional methods and genotypic characterization was carried out using 16S r-DNA partial gene amplification and sequencing. The broken down by products of DNT in the extract was determined by Gas Chromatogram- Mass spectrometry (GC-MS). Supernatant samples from the broth studied at 24 h interval were analyzed by HPLC analysis and the effect on various nutritional and environmental factors were analysed and optimized for the isolate. Results: Out of three isolates one strain TUHP2 were found to have potent efficiency to degrade TNT and revealed the genus Bacillus. 16S rDNA gene sequence analysis showed highest homology (98%) with Bacillus cereus and was assigned as Bacillus cereus sp TUHP2. Based on the energy of the predicted models, the secondary structure predicted by MFE showed the more stable structure with a minimum energy. Products of TNT Transformation showed colour change in the medium during cultivation. TNT derivates such as 2HADNT and 4HADNT were detected by HPLC chromatogram and 2ADNT, 4ADNT by GC/MS analysis. Conclusion: Hence this study presents the clear evidence for the biodegradation process of TNT by strain Bacillus cereus sp TUHP2.

Keywords: bioremediation, biodegradation, biotransformation, sequencing

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Authors: Zehour Rahmani, Messouda Dekmouche, Mohamed Hadjadj, Mokhtar Saidi

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In the last decades of the nineteenth century, the study of disease – causing microorganisms became concentrated on bacteria and largely institutionalized. In earlier years, the scientists interested in bacteria had originally been chemists like Pasteur, physicists like Tyndall, or botanists like Cohn and ward. For this reason, the objective of this research was to evaluate the potential of some dithiolethiones on standard microorganism strains as well as multi-drug resistant bacteria, which were isolated from hospitals. Recent studies have demonstrated, that several dithiolethione compounds, particularly (3H-1,2-dithiole-3-thione), exhibit the biological activities against several bacteria.

Keywords: bacteria, dithiolethiones, microorganism, potential

Procedia PDF Downloads 284

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

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Authors: Hai Chi, Ibrahim Mehmeti, Kirill Ovchinnikov, Hegle Holo, Ingolf F. Nes, Dzung B. Diep

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By using a panel of multiple indicator strains of different bacterial species and genera, we screened a large collection of bacterial isolates (over 1800 isolates) derived from raw milk, for bacteriocin producers with broad inhibition spectra (BIS). Fourteen isolates with BIS were identified, and by 16S rDNA sequencing they were found to belong to Lactococcus garvieae (10 isolates) and Enterococcus feacalis (4 isolates). Further analysis of the ten L. garvieae isolates revealed that they were very similar, if not identical, to each other in metabolic and genetic terms: they had the same fermentation profile on different types of sugars, repetitive sequence-based PCR (rep-PCR) DNA pattern as well as they all had the same inhibition profile towards over 50 isolates of different species. The bacteriocin activity from one of the L. garvieae isolates was assessed further. The bacteriocin which was termed garvicin KS, was found to be heatstable and proteinase-labile and its inhibition spectrum contained many distantly related genera of Firmicutes, comprising most lactic acid bacteria (LAB) as well as problematic species of Bacillus, Listeria, Streptococcus and Staphylococcus and their antibiotic resistant derivatives (e.g. VRE, MRSA). Taken together, the results indicate that this is a potent bacteriocin from L. garvieae and that its very broad inhibition spectrum can be a very useful property for use in food preservation as well as in infection treatments caused by gram-positive pathogens and their antibiotic-derivatives.

Keywords: bacteriocin, lactic acid bacteria, Lactococcus garvieae, antibiotics resistance

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Authors: Eusebio A. Jiménez-Arévalo, Deifilia Ahuatzi-Chacón, Juvencio Galíndez-Mayer, Cleotilde Juárez-Ramírez, Nora Ruiz-Ordaz

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Bendiocarb is a known toxic xenobiotic that presents acute and chronic risks for freshwater invertebrates and estuarine and marine biota; thus, the treatment of water contaminated with the insecticide is of concern. In this paper, a bacterial community with the capacity to grow in bendiocarb as its sole carbon and nitrogen source was isolated by enrichment techniques in batch culture, from samples of a composting plant located in the northeast of Mexico City. Eight cultivable bacteria were isolated from the microbial community, by PCR amplification of 16 rDNA; Pseudoxanthomonas spadix (NC_016147.2, 98%), Ochrobacterium anthropi (NC_009668.1, 97%), Staphylococcus capitis (NZ_CP007601.1, 99%), Bosea thiooxidans. (NZ_LMAR01000067.1, 99%), Pseudomonas denitrificans. (NC_020829.1, 99%), Agromyces sp. (NZ_LMKQ01000001.1, 98%), Bacillus thuringiensis. (NC_022873.1, 97%), Pseudomonas alkylphenolia (NZ_CP009048.1, 98%). NCBI accession numbers and percentage of similarity are indicated in parentheses. These bacteria were regarded as the isolated species for having the best similarity matches. The ability to degrade bendiocarb by the immobilized bacterial community in a packed bed biofilm reactor, using as support volcanic stone fragments (tezontle), was evaluated. The reactor system was operated in batch using mineral salts medium and 30 mg/L of bendiocarb as carbon and nitrogen source. With this system, an overall removal efficiency (η_bend) rounding 90%, was reached.

Keywords: bendiocarb, biodegradation, biofilm reactor, carbamate insecticide

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Authors: Mehani Mouna, Ladjel segni

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Humans use plants for thousands of years to treat various ailments, In many developing countries, Much of the population relies on traditional doctors and their collections of medicinal plants to cure them. Essential oils have many therapeutic properties. In herbal medicine, They are used for their antiseptic properties against infectious diseases of fungal origin, Against dermatophytes, Those of bacterial origin. The aim of our study is to determine the antimicrobial effect of essential oils of the plant Schinus molle on some pathogenic bacteria. It is a medicinal plant used in traditional therapy. Essential oils have many therapeutic properties. In herbal medicine, They are used for their antiseptic properties against infectious diseases of fungal origin, Against dermatophytes, Those of bacterial origin. The test adopted is based on the diffusion method on solid medium (Antibiogram), This method allows to determine the susceptibility or resistance of an organism according to the sample studied. Our study reveals that the essential oil of the plant Schinus molle has a different effect on the resistance of germs: For Pseudomonas aeruginosa strain is a moderately sensitive with an inhibition zone of 10 mm, Further Antirobactere, Escherichia coli and Proteus are strains that represent a high sensitivity, A zone of inhibition equal to 14.66 mm.

Keywords: Essential oil, microorganism, antibiogram, shinus molle

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Authors: Kubilay Kurtulus Bastas

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Bacterial diseases are very destructive and cause economic losses on pears. Promising plant extracts for the management of plant diseases are environmentally safe, long-lasting and extracts of certain plants contain alkaloids, tannins, quinones, coumarins, phenolic compounds, and phytoalexins. In this study, bacteria were isolated from different parts of pear exhibiting characteristic symptoms of bacterial diseases from the Central Anatolia, Turkey. Pathogenic bacteria were identified by morphological, physiological, biochemical and molecular methods as fire blight (Erwinia amylovora (39%)), bacterial blossom blast and blister bark (Pseudomonas syringae pv. syringae (22%)), crown gall (Rhizobium radiobacter (1%)) from different pear cultivars, and determined virulence levels of the pathogens with pathogenicity tests. The air-dried 25 plant material was ground into fine powder and extraction was performed at room temperature by maceration with 80% (v/v) methanol/distilled water. The minimum inhibitory concentration (MIC) values were determined by using modified disc diffusion method at five different concentrations and streptomycin sulphate was used as control chemical. Bacterial suspensions were prepared as 108 CFU ml⁻¹ densities and 100 µl bacterial suspensions were spread to TSA medium. Antimicrobial activity was evaluated by measuring the inhibition zones in reference to the test organisms. Among the tested plants, Origanum vulgare, Hedera helix, Satureja hortensis, Rhus coriaria, Eucalyptus globulus, Rosmarinus officinalis, Ocimum basilicum, Salvia officinalis, Cuminum cyminum and Thymus vulgaris showed a good antibacterial activity and they inhibited the growth of the pathogens with inhibition zone diameter ranging from 7 to 27 mm at 20% (w/v) in absolute methanol in vitro conditions. In vivo, the highest efficacy was determined as 27% on reducing tumor formation of R. radiobacter, and 48% and 41% on reducing shoot blight of E. amylovora and P. s. pv. syringae on pear seedlings, respectively. Obtaining data indicated that some plant extracts may be used against the bacterial diseases on pome fruits within sustainable and organic management programs.

Keywords: bacteria, eco-friendly management, organic, pear, plant extract

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Authors: Bassam O AlJohny

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The growth of Bacillus aquimaris was inhibited from 6 - 20 % of NaCl but it showed some tolerance when Diatomaceous earth (DE) added from 2 - 12% NaCl. Concerning the effect of NaCl on polyol production, we can conclude that, the test bacterium showed some tolerance to NaCl by producing glycerol up to 8 % of NaCl. Then decreased sharply. The addition of DE decrease the amount of polyol and glycerol remarkably and this due to the productive effect of DE to the bacterial cells. The SEM figures represented the presence of electron dense bodies due to the accumulation of small particles of DE as protective molecules.

Keywords: Bacillus aquimaris, Diatomaceous earth (DE), osmoticstress, sodium chloride

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Authors: Sachin Kamble, Shradha Gawankar

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This paper presents the findings of successful implementation of Business Process Reengineering (BPR) of cement dispatch activities in a cement manufacturing plant located in India. Simulation model was developed for the purpose of identifying and analyzing the areas for improvement. The company was facing a problem of low throughput rate and subsequent forced stoppages of the plant leading to a high production loss of 15000MT per month. It was found from the study that the present systems and procedures related to the in-plant logistics plant required significant changes. The major recommendations included process improvement at the entry gate, reducing the cycle time at the security gate and installation of an additional weigh bridge. This paper demonstrates how BPR can be implemented for improving the in-plant logistics process. Various recommendations helped the plant to increase its throughput by 14%.

Keywords: in-plant logistics, cement logistics, simulation modelling, business process re-engineering, supply chain management

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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: Narasimhulu Korrapati

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The objective of this present study is to optimize the process parameters for batch biosorption of Cr(VI) and Cu(II) ions by Bacillus subtilis using Response Surface Methodology (RSM). Batch biosorption studies were conducted under optimum pH, temperature, biomass concentration and contact time for the removal of Cr(VI) and Cu(II) ions using Bacillus subtilis. From the studies it is noticed that the maximum biosorption of Cr(VI) and Cu(II) was by Bacillus subtilis at optimum conditions of contact time of 30 minutes, pH of 4.0, biomass concentration of 2.0 mg/mL, the temperature of 32°C in batch biosorption studies. Predicted percent biosorption of the selected heavy metal ions by the design expert software is in agreement with experimental results of percent biosorption. The percent biosorption of Cr(VI) and Cu(II) in batch studies is 80% and 78.4%, respectively.

Keywords: heavy metal ions, response surface methodology, biosorption, wastewater

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Authors: Emmanuel Ekpa, Adaji Andrew, Queen Opaluwa, Isreal Daraobong

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Three (3) indigenous bacteria species (Bacillus spp, Acinectobacter spp and Moraxella spp) previously isolated from contaminated soil of some auto mechanic workshops were used for bioremediation studies on some irrigation water used at Sarkin-noma Fadama farms located in Lokoja Kogi State, Nigeria. This was done in order to investigate their bioremediation potentials using a simple pour plate method. The physicochemical parameters and heavy metal analysis (using AAS iCE 3000) of the irrigation water were performed before and after inoculation of the isolated organisms. Nitrate and phosphate concentration were found to be 10.56mg/L and 12.63mg/L prior to inoculation while iron and zinc were 0.9569mg/L and 0.2245mg/L respectively. Other physicochemical parameters were also observed to be high prior to inoculation. After the bioremediation test (inoculation with the isolated organisms), a nitrate and phosphate content of 2.53mg/L and 2.61mg/L were recorded respectively, iron and zinc gave 0.1694mg/L and 0.0174mg/L concentrations while other physicochemical parameters measured were also found to be lower in their respective values. The implication of this present study is that a number of carefully isolated indigenous bacteria species are capable of reducing the amount of heavy metal concentrations in water. Also, non-metallic contaminants like nitrate and phosphate are susceptible to bioremediation in the presence of such efficient system.

Keywords: bioremediation, heavy metals, physicochemical parameters, Bacillus spp, Acinectobacter spp and Moraxella spp, AAS, spectrometer 3000

Procedia PDF Downloads 304

Authors: K. Petrosyan, R. Piwowarczyk, K. Ruraż, S. Thijs, J. Vangronsveld, W. Kaca

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Holoparasitic plants are flowering heterotrophic angiosperms which with the help of an absorbing organ - haustorium, attach to another plant, the so-called the host. Due to the different hosts, unusual lifestyle, lack of roots, chlorophylls and photosynthesis, these plants are interesting and unique study objects for global biodiversity. The seeds germination of the parasitic plants also is unique: they germinate only in response to germination stimulants, namely strigolactones produced by the root of an appropriate host. Resistance of the seeds on different environmental conditions allow them to stay viable in the soil for more than 20 years. Among the wide range of plant protection mechanisms the endophytic communities have a specific role. In this way, they have the potential to mitigate the impacts of adverse conditions such as soil salinization. The major objective of our study was to compare the bacterial endo-microbiomes from seeds of two holoparasitic plants from Orobanchaceae family, Cistanche – C. armena (Armenia) and C. phelypaea (Portugal) – from saline habitats different in soil water status. The research aimed to perform how environmental conditions influence on the diversity of the bacterial communities of C. armena and C. phelypaea seeds. This was achieved by comparison of the endophytic microbiomes of two species and isolation of culturable bacteria. A combination of culture-dependent and molecular techniques was employed for the identification of the seed endomicrobiome (culturable and unculturable). Using the V3-V4 hypervariable region of the 16S rRNA gene, four main taxa were identified: Proteobacteria, Actinobacteria, Bacteroidetes, Firmicutes, but the relative proportion of the taxa was different in each type of seed. Generally, sixteen phyla, 323 genera and 710 bacterial species were identified, mainly Gram negative, halotolerant bacteria with an environmental origin. However, also some unclassified and unexplored taxonomic groups were found in the seeds of both plants. 16S rRNA gene sequencing analysis from both species identified the gram positive, endospore forming, halotolerant and alkaliphile Bacillus spp. which suggests that the endophytic bacteria of examined seeds possess traits that are correlated with the natural habitat of their hosts. The cultivable seed endophytes from C. armena and C. phelypaea were rather similar, notwithstanding the big distances between their growth habitats - Armenia and Portugal. Although the seed endophytic microbiomes of C. armena and C. phelypaea contain a high number of common bacterial taxa, also remarkable differences exist. We demonstrated that the environmental conditions or abiotic stresses influence on diversity of the bacterial communities of holoparasiotic seeds. To the best of our knowledge the research is the first report of endophytes from seeds of holoparasitic Cistanche armena and C. phelypaea plants.

Keywords: microbiome, parasitic plant, salinity, seeds

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Authors: Uftah Ali M. Shushni, Viola Stuppec, Ulrike Lindequist

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Because of the evolving resistance of microorganisms to existing antibiotics, there is an increasing need for new antibiotics not only in human but also in veterinary medicine. As part of our ongoing work on the secondary metabolites produced by marine fungi, the organic extract of the culture filtrate of an Ascomycetous fungus, which was found on driftwood collected from the coast of the Greifswalder Bodden, Baltic Sea, Germany displayed antimicrobial activity against some fish and human pathogenic bacteria. Bioactivity-guided column chromatographic separation led to the isolation of 6-Deoxybostrycoidin. The structure was determined from the interpretation of spectroscopic data (UV, MS, and NMR). 6-Deoxybostrycoidin exhibited in vitro activity against Bacillus subtilis, Staphylococcus aureus and Flexibacter maritimus with minimal inhibitory concentrations of 25, 12.5 and 12.5 μg/ml respectively.

Keywords: marine fungi, fish pathogenic bacteria, microorganism, medicine

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Authors: Kraiem Khadija, Hamadi Kallali, Naceur Jedidi

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Constructed wetland systems are eco-technology recognized as environmentally friendly and emerging innovative solutions remediation as these systems are cost-effective and sustainable wastewater treatment systems. The performance of these biological system is affected by various factors such as plant, substrate, wastewater type, hydraulic loading rate, hydraulic retention time, water depth, and operation mood. The objective of this study was to to assess the alters of plant species on pollutants reduction and enrichment of anammox and nitrifing denitrifing bacteria in a modified vertical flow (VFCW) constructed wetland. This tests were carried out using three modified vertical constructed wetlands with a surface of 0.23 m² and depth 80 cm. It was a saturated vertical constructed wetland at the bottom. The saturation zone is maintained by the siphon structure at the outlet. The VFCW (₁) system was unplanted, VFCW (₂) planted with Typha angustofolia, and VFCW(₃) planted with Phragmites australis. The experimental units were fed with domestic wastewater and were operated by batch mode during 8 months at an average hydraulic loading rate around 20 cm day− 1. The operation cycle was two days feeding and five days rest. Results indicated that plants presence improved the removal efficiency; the removal rates of organic matter (85.1–90.9%; COD and 81.8–88.9%; BOD5), nitrogen (54.2–73%; NTK and 66–77%; NH4 -N) were higher by 10.7–30.1% compared to the unplanted vertical constructed wetland. On the other hand, the plant species had no significant effect on removal efficiency of COD, The removal of COD was similar in VFCW (₂) and VFCW (₃) (p > 0.05), attaining average removal efficiencies of 88.7% and 85.2%, respectively. Whereas it had a significant effect on NTK removal (p > 0.05), with an average removal rate of 72% versus 51% for VFCW (₂) and VFCW (₃), respectively. Among the three sets of vertical flow constructed wetlands, the VFCW(₂) removed the highest percent of total streptococcus, fecal streptococcus total coliforms, fecal coliforms, E. coli as 59, 62, 52, 63, and 58%, respectively. The presence and the plant species alters the community composition and abundance of the bacteria. The abundance of bacteria in the planted wetland was much higher than that in the unplanted one. VFCW(₃) had the highest relative abundance of nitrifying bacteria such as Nitrosospira (18%), Nitrosospira (12%), and Nitrobacter (8%). Whereas the vertical constructed wetland planted with typha had larger number of denitrifying species, with relative abundances of Aeromonas (13%), Paracoccus (11%), Thauera (7%), and Thiobacillus (6%). However, the abundance of nitrifying bacteria was very lower in this system than VFCW(₂). Interestingly, the presence of Thypha angustofolia species favored the enrichment of anammox bacteria compared to unplanted system and system planted with phragmites australis. The results showed that the middle layer had the most accumulation of anammox bacteria, which the anaerobic condition is better and the root system is moderate. Vegetation has several characteristics that make it an essential component of wetlands, but its exact effects are complex and debated.

Keywords: wastawater, constructed wetland, anammox, removal

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Authors: T. Deepa, S. R. Inchara, S. V. Venkatesh, Seema Tharannum

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Concrete is the most widely used structural material. It is made using locally available materials. However, Concrete has low tensile strength and may crack in the early days with exothermic hydration. Bacillus subtilis bacteria that form endospores is the biological agent considered in this study for Biomineralization or MICP (Microbially Induced Calcite Precipitation) Technique and to address the increased Construction water demand, Recycled Grey Water which is obtained from STP of PES University, opted in place of Potable water. In this work, M30 grade conventional concrete is designed using OPC 53 grade cement, Manufactured Sand, Natural coarse aggregates, and Potable water. Conventional Concrete (CC), Bacterial Concrete with Potable water (BS), and Recycled Grey Water concrete (RGW) are the three different concrete specimens casted. Experimental studies such as the strength test and the surface hardness test are conducted on Conventional and Bacterial concrete samples after 7, 28, and 56 days of curing. Concrete cubes are subjected to a temperature of 50° C to investigate the effect of higher temperature. Cracked cube specimens are observed for Self-healing - as well as microstructure analysis with Scanning Electron Microscope (SEM), Energy Dispersive X-Ray Analysis (EDAX), and X-Ray Diffraction Analysis (XRD).Noticeable Calcium salt deposition is observed on the surface of BS and RGW cracked specimen. Surface hardness and EDAX test gave promising result on the advantage of using spore-forming bacteria in concrete. This is followed by the strength gain in Compression and Flexure. Results also indicate that Recycled Grey Water can be a substitute for Normal water in concrete.

Keywords: bacillus subtilis, bacterial concrete, recycled grey water, self-healing, surface hardness of concrete

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Authors: Aurora Gitto, Philipp Proksch

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The presence of various microorganisms (bacteria, fungi) in surface water and groundwater represents an important issue for human health worldwide. The matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) has emerged as a promising and reliable tool for bacteria identification in clinical diagnostic microbiology and environmental strains thanks to an ionization technique that uses a laser energy absorbing matrix to create ions from large molecules with minimal fragmentation. The study aims first to conceptualise and set up library information and create a comprehensive database of MALDI-TOF-MS spectra from environmental water samples. The samples were analysed over a year (2021-2022) using membrane filtration methodology (0.45 μm and 0.22 μm) and then isolated on R2A agar for a period of 5 days and Yeast extract agar growing at 22 °C up to 4 days and 37 °C for 48 hours. The undetected organisms by MALDI-TOF-MS were analysed by PCR and then sequenced. The information obtained by the sequencing was further implemented in the MALDI-TOF-MS library. Among the culturable bacteria, the results show how the incubator temperature affects the growth of some genera instead of others, as demonstrated by Pseudomonas sp., which grows at 22 °C, compared to Bacillus sp., which is abundant at 37 °C. The bacteria community shows a variation in composition also between the media used, as demonstrated with R2A agar which has been defined by a higher presence of organisms not detected compared to YEA. Interesting is the variability of the Genus over one year of sampling and how the seasonality impacts the bacteria community; in fact, in some sampling locations, we observed how the composition changed, moving from winter to spring and summer. In conclusion, the bacteria community in groundwater and river bank filtration represents important information that needs to be added to the library to simplify future water quality analysis but mainly to prevent potential risks to human health.

Keywords: water quality, MALDI-TOF-MS, sequencing, library

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