Search results for: microbial property

Authors: Goksen Arik, Mihriban Korukluoglu

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Lactic acid bacteria (LAB) has been used commonly in the food industry. They can be used as natural preservatives because acidifying carried out in the medium can protect the last product against microbial spoilage. Besides, other metabolites produced by LAB during fermentation period have also an antimicrobial effect on pathogen and spoilage microorganisms in the food industry. LAB are responsible for the desirable and distinctive aroma and flavour which are observed in fermented food products such as pickle, kefir, yogurt, and cheese. Various LAB strains are able to produce B-group vitamins such as folate (B11), riboflavin (B2) and cobalamin (B12). Especially wild-type strains of LAB can produce B-group vitamins in high concentrations. These cultures may be used in food industry as a starter culture and also the microbial strains can be used in encapsulation technology for new and functional food product development. This review is based on the current applications of B-group vitamin producing LAB. Furthermore, the new technologies and innovative researches about B vitamin production in LAB have been demonstrated and discussed for determining their usage availability in various area in the food industry.

Keywords: B vitamin, food industry, lactic acid bacteria, starter culture, technology

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Authors: Y. M. Al-Wahaibi, S. N. Al-Bahry, A. E. Elshafie, A. S. Al-Bemani, S. J. Joshi, A. K. Al-Bahri

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Crude oil is a major source of global energy. The major problem is its widespread use and demand resulted is in increasing environmental pollution. One associated pollution problem is ‘oil spills’. Oil spills can be remediated with the use of chemical dispersants, microbial biodegradation and microbial metabolites such as biosurfactants. Four different minimal salt media for biosurfactant production by Bacillus isolated from oil contaminated sites from Oman were screened. These minimal salt media were supplemented with either glucose or sucrose as a carbon source. Among the isolates, W16 and B30 produced the most active biosurfactants. Isolate W16 produced better biosurfactant than the rest, and reduced surface tension (ST) and interfacial tension (IFT) to 25.26mN/m and 2.29mN/m respectively within 48h which are characteristics for removal of oil in contaminated sites. Biosurfactant was produced in bulk and extracted using acid precipitation method. Thin Layer Chromatography (TLC) of acid precipitate biosurfactant revealed two concentrated bands. Further studies of W16 biosurfactant in bioremediation of oil spills are recommended.

Keywords: oil contamination, remediation, Bacillus spp, biosurfactant, surface tension, interfacial tension

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Authors: Velvizhi Dharmalingam, Rajalaksmi Ramalingam, Rekha Prabhu, Ilavarasan Raju

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Background: The use of herbal products for various therapeutic regimens has increased tremendously in the developing countries. Elaeocarpus ganitrus(Rudraksha) is a broad-leaved tree, belonging to the family Elaeocarpaceae found in tropical and subtropical areas. It is popular in an indigenous system of medicine like Ayurveda, Siddha, and Unani. According to Ayurvedic medicine, Rudraksha is used in the managing of blood pressure, asthma, mental disorders, diabetes, gynaecological disorders, neurological disorders such as epilepsy and liver diseases. Objectives: The present study aimed to study the physicochemical parameters of Elaeocarpus ganitrus(fruits) and identify the phenolic compounds (gallic acid, ellagic acid, and chebulinic acid). To estimate the microbial load and the antibacterial activity of extract of Elaeocarpus ganitrus for selective pathogens. Methodology: The dried powdered fruit of Elaeocarpus ganitrus was performed the physicochemical parameters (such as Loss on drying, Alcohol soluble extractive, Water soluble extractive, Total ash and Acid insoluble ash) and pH was measured. The dried coarse powdered fruit of Elaeocarpus ganitrus was extracted successively with hexane, chloroform, ethylacetate and aqueous alcohol by cold percolation method. Identification of phenolic compounds (gallic acid, ellagic acid, chebulinic acid) was done by HPTLC method and confirmed by co-TLC using different solvent system.The successive extracts of Elaeocarpus ganitrus and standards (like gallic acid, ellagic acid, and chebulinic acid) was approximately weighed and made up with alcohol. HPTLC (CAMAG) analysis was performed on a TLC over silica gel 60F254 precoated aluminium plate, layer thickness 0.2 mm (E.Merck, Germany) by using ATS4, Visualizer and Scanner with wavelength at 254 nm, 366 nm and derivatized with different reagents. The microbial load such as total bacterial count, total fungal count, Enterobacteria, Escherichia coli, Salmonella species, Staphylococcus aureus and Pseudomonas aeruginosa by serial dilution method and antibacterial activity of was measured by Kirby bauer method for selective pathogens. Results: The physicochemical parameter of Elaeocarpus ganitrus was studied for standardization of crude drug. Among all the successive extracts were identified with phenolic compounds and Elaeocarpus ganitrus extract having potent antibacterial activity against gram-positive and gram-negative bacteria.

Keywords: antimicrobial activity, Elaeocarpus ganitrus, HPTLC, phenolic compounds

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Authors: I. Nava-Arenas, N. Ruiz-Ordaz, C. J. Galindez-Mayer, M. L. Luna-Guido, S. L. Ruiz-López, A. Cabrera-Orozco, D. Nava-Arenas

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Among the most important herbicides, the organochlorine compounds are of considerable interest due to their recalcitrance to the chemical, biological, and photolytic degradation, their persistence in the environment, their mobility, and their bioacummulation. The most widely used herbicides in North America are primarily 2,4-dichlorophenoxyacetic acid (2,4-D), the triazines (atrazine and simazine), and to a lesser extent diuron. The contamination of soils and water bodies frequently occurs by mixtures of these xenobiotics. For this reason, in this work, the operational stability to changes in the composition of the medium supplied to an aerobic biofilm reactor was studied. The reactor was packed with fragments of volcanic rock that retained a complex microbial film, able to degrade a mixture of organochlorine herbicides atrazine, simazine, diuron and 2,4-D, and whose members have microbial genes encoding the main catabolic enzymes atzABCD, tfdACD and puhB. To acclimate the attached microbial community, the biofilm reactor was fed continuously with a mineral minimal medium containing the herbicides (in mg•L-1): diuron, 20.4; atrazine, 14.2, simazine, 11.4, and 2,4-D, 59.7, as carbon and nitrogen sources. Throughout the bioprocess, removal efficiencies of 92-100% for herbicides, 78-90% for COD, 92-96% for TOC and 61-83% for dehalogenation were reached. In the microbial community, the genes encoding catabolic enzymes of different herbicides tfdACD, puhB and, occasionally, the genes atzA and atzC were detected. After the acclimatization, the triazine herbicides were eliminated from the mixture formulation. Volumetric loading rates of the mixture 2,4-D and diuron were continuously supplied to the reactor (1.9-21.5 mg herbicides •L-1 •h-1). Along the bioprocess, the removal efficiencies obtained were 86-100% for the mixture of herbicides, 63-94% for for COD, 90-100% for COT, and dehalogenation values of 63-100%. It was also observed that the genes encoding the enzymes in the catabolism of both herbicides, tfdACD and puhB, were consistently detected; and, occasionally, the atzA and atzC. Subsequently, the triazine herbicide atrazine and simazine were restored to the medium supply. Different volumetric charges of this mixture were continuously fed to the reactor (2.9 to 12.6 mg herbicides •L-1 •h-1). During this new treatment process, removal efficiencies of 65-95% for the mixture of herbicides, 63-92% for COD, 66-89% for TOC and 73-94% of dehalogenation were observed. In this last case, the genes tfdACD, puhB and atzABC encoding for the enzymes involved in the catabolism of the distinct herbicides were consistently detected. The atzD gene, encoding the cyanuric hydrolase enzyme, could not be detected, though it was determined that there was partial degradation of cyanuric acid. In general, the community in the biofilm reactor showed some catabolic stability, adapting to changes in loading rates and composition of the mixture of herbicides, and preserving their ability to degrade the four herbicides tested; although, there was a significant delay in the response time to recover to degradation of the herbicides.

Keywords: biodegradation, biofilm reactor, microbial community, organochlorine herbicides

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Authors: Payel Ghosh, Rama Chandra Pradhan, Sabyasachi Mishra

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Jamun (Syzygium cuminii L.) is one of the important indigenous minor fruit with high medicinal value. The jamun cultivation is unorganized and there is huge loss of this fruit every year. The perishable nature of the fruit makes its postharvest management further difficult. Due to the strong cell wall structure of pectin-protein bonds and hard seeds, extraction of juice becomes difficult. Enzymatic treatment has been commercially used for improvement of juice quality with high yield. The objective of the study was to optimize the best treatment method for juice extraction. Enzymes (Pectinase and Tannase) from different stains had been used and for each enzyme, best result obtained by using response surface methodology. Optimization had been done on the basis of physicochemical property, nutritional property, sensory quality and cost estimation. According to quality aspect, cost analysis and sensory evaluation, the optimizing enzymatic treatment was obtained by Pectinase from Aspergillus aculeatus strain. The optimum condition for the treatment was 44 ^oC with 80 minute with a concentration of 0.05% (w/w). At these conditions, 75% of yield with turbidity of 32.21NTU, clarity of 74.39%T, polyphenol content of 115.31 mg GAE/g, protein content of 102.43 mg/g have been obtained with a significant difference in overall acceptability.

Keywords: enzymatic treatment, Jamun, optimization, physicochemical property, sensory analysis

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Authors: Umar Yusuf Batsari

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Let E be a uniformly smooth and uniformly convex real Banach space and C be a nonempty, closed and convex subset of E. Let $V= \{S_i : C\to C, ~i=1, 2, 3\cdots N\}$ be a convex set of relatively nonexpansive mappings containing identity. In this paper, an iterative sequence obtained from CQ algorithm was shown to have strongly converge to a point $\hat{x}$ which is a common fixed point of relatively nonexpansive mappings in V and also solve the system of equilibrium problems in E. The result improve some existing results in the literature.

Keywords: relatively nonexpansive mappings, strong convergence, equilibrium problems, uniformly smooth space, uniformly convex space, convex set, kadec klee property

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Authors: Lana Horvat Dmitrović

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The main purpose of this paper is to study the box dimension as fractal property of bifurcations of discrete dynamical systems in higher dimensions. The paper contains the fractal analysis of the orbits near the hyperbolic and non-hyperbolic fixed points in discrete dynamical systems. It is already known that in one-dimensional case the orbit near the hyperbolic fixed point has the box dimension equal to zero. On the other hand, the orbit near the non-hyperbolic fixed point has strictly positive box dimension which is connected to the non-degeneracy condition of certain bifurcation. One of the main results in this paper is the generalisation of results about box dimension near the hyperbolic and non-hyperbolic fixed points to higher dimensions. In the process of determining box dimension, the restriction of systems to stable, unstable and center manifolds, Lipschitz property of box dimension and the notion of projective box dimension are used. The analysis of the bifurcations in higher dimensions with one multiplier on the unit circle is done by using the normal forms on one-dimensional center manifolds. This specific change in box dimension of an orbit at the moment of bifurcation has already been explored for some bifurcations in one and two dimensions. It was shown that specific values of box dimension are connected to appropriate bifurcations such as fold, flip, cusp or Neimark-Sacker bifurcation. This paper further explores this connection of box dimension as fractal property to some specific bifurcations in higher dimensions, such as fold-flip and flip-Neimark-Sacker. Furthermore, the application of the results to the unit time map of continuous dynamical system near hyperbolic and non-hyperbolic singularities is presented. In that way, box dimensions which are specific for certain bifurcations of continuous systems can be obtained. The approach to bifurcation analysis by using the box dimension as specific fractal property of orbits can lead to better understanding of bifurcation phenomenon. It could also be useful in detecting the existence or nonexistence of bifurcations of discrete and continuous dynamical systems.

Keywords: bifurcation, box dimension, invariant manifold, orbit near fixed point

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Authors: Neda Lovričević, Đilda Pečarić, Josip Pečarić

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The Jensen functional in its discrete form is brought in relation to the Csiszar divergence functional, this time via its monotonicity property. This approach presents a generalization of the previously obtained results that made use of interpolating Jensen-type inequalities. Thus the monotonicity property is integrated with the Zipf-Mandelbrot law and applied to f-divergences for probability distributions that originate from the Csiszar divergence functional: Kullback-Leibler divergence, Hellinger distance, Bhattacharyya distance, chi-square divergence, total variation distance. The Zipf-Mandelbrot and the Zipf law are widely used in various scientific fields and interdisciplinary and here the focus is on the aspect of the mathematical inequalities.

Keywords: Jensen functional, monotonicity, Csiszar divergence functional, f-divergences, Zipf-Mandelbrot law

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Authors: Suganya Chandrababu, Dhundy Bastola

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Like in the gut, the subgingival microbiota plays a crucial role in oral hygiene, health, and cariogenic diseases. Human activities like diet, antibiotics, and periodontal treatments alter the bacterial communities, metabolism, and functions in the oral cavity, leading to a dysbiotic state and changes in the plaques of orthodontic patients. Fixed periodontal appliances hinder oral hygiene and cause changes in the dental plaques influencing the subgingival microbiota. However, the microbial species’ diversity and complexity pose a great challenge in understanding the taxa’s community distribution patterns and their role in oral health. In this research, we analyze the subgingival microbial samples from individuals with fixed dental appliances (metal/clear) using an in silico approach. We employ exploratory hypothesis-driven multivariate and regression analysis to shed light on the microbial community and its functional fluctuations due to dental appliances used and identify risks associated with complex disease phenotypes. Our findings confirm the changes in oral microbiota composition due to the presence and type of fixed orthodontal devices. We identified seven main periodontic pathogens, including Bacteroidetes, Actinobacteria, Proteobacteria, Fusobacteria, and Firmicutes, whose abundances were significantly altered due to the presence and type of fixed appliances used. In the case of metal braces, the abundances of Bacteroidetes, Proteobacteria, Fusobacteria, Candidatus saccharibacteria, and Spirochaetes significantly increased, while the abundance of Firmicutes and Actinobacteria decreased. However, in individuals With clear braces, the abundance of Bacteroidetes and Candidatus saccharibacteria increased. The highest abundance value (P-value=0.004 < 0.05) was observed with Bacteroidetes in individuals with the metal appliance, which is associated with gingivitis, periodontitis, endodontic infections, and odontogenic abscesses. Overall, the bacterial abundances decrease with clear type and increase with metal type of braces. Regression analysis further validated the multivariate analysis of variance (MANOVA) results, supporting the hypothesis that the presence and type of the fixed oral appliances significantly alter the bacterial abundance and composition.

Keywords: oral microbiota, statistical analysis, fixed or-thodontal appliances, bacterial abundance, multivariate analysis, regression analysis

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Authors: Yixin Yan, Miao Yan, Irini Angelidaki, Ioannis Fotidis

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Ammonia, which derives from the degradation of urea and protein-substrates, is the major toxicant of the commercial anaerobic digestion reactors causing loses of up to 1/3 of their practical biogas production, which reflects directly on the overall revenue of the plants. The current experimental work is aiming to alleviate the ammonia inhibition in anaerobic digestion (AD) process by developing an innovative bioaugmentation method of ammonia tolerant methanogenic consortia. The ammonia tolerant consortia were cultured in batch reactors and immobilized together with biochar in agar (customized inocula). Three continuous stirred-tank reactors (CSTR), fed with the organic fraction of municipal solid waste at a hydraulic retention time of 15 days and operated at thermophilic (55°C) conditions were assessed. After an ammonia shock of 4 g NH4+-N L-1, the customized inocula were bioaugmented into the CSTR reactors to alleviate ammonia toxicity effect on AD process. Recovery rate of methane production and methanogenic activity will be assessed to evaluate the bioaugmentation performance, while 16s rRNA gene sequence will be used to reveal the difference of microbial community changes through bioaugmentation. At the microbial level, the microbial community structures of the four reactors will be analysed to find the mechanism of bioaugmentation. Changes in hydrogen formation potential will be used to predict direct interspecies electron transfer (DIET) between ammonia tolerant methanogens and syntrophic bacteria. This experimental work is expected to create bioaugmentation inocula that will be easy to obtain, transport, handled and bioaugment in AD reactors to efficiently alleviate the ammonia toxicity, without alternating any of the other operational parameters including the ammonia-rich feedstocks.

Keywords: artisanal fishing waste, acidogenesis, volatile fatty acids, pH, inoculum/substrate ratio

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Authors: Omaima A. Sharaf, Tarek A. Moussa, Said M. Badr El-Din, H. Moawad

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Background: Polycyclic aromatic hydrocarbons (PAHs) pose great environmental and human health concerns for their widespread occurrence, persistence, and carcinogenic properties. PAHs releases due to anthropogenic activities to the wider environment have led to higher concentrations of these contaminants than would be expected from natural processes alone. This may result in a wide range of environmental problems that can accumulate in agricultural ecosystems, which threatened to become a negative impact on sustainable agricultural development. Thus, this study aimed to evaluate the physico-chemical, and microbial properties of the compost leachate (CL) to assess its role as nutrient and microbial source (biostimulation/bioaugmentation) for developing a cost-effective bioremediation technology for PAHs contaminated sites. Material and Methods: PAHs-degrading bacteria were isolated from CL that was collected from a composting site located in central Scotland, UK. Isolation was carried out by enrichment using phenanthrene (PHR), pyrene (PYR) and benzo(a)pyrene (BaP) as the sole source of carbon and energy. The isolates were characterized using a variety of phenotypic and molecular properties. Six different isolates were identified based on the difference in morphological and biochemical tests. The efficiency of these isolates in PAHs utilization was assessed. Further analysis was performed to define taxonomical status and phylogenic relation between the most potent PAHs-utilizing bacterial strains and other standard strains, using molecular approach by partial 16S rDNA gene sequence analysis. Results indicated that the 16S rDNA sequence analysis confirmed the results of biochemical identification, as both of biochemical and molecular identification of the isolates assigned them to Bacillus licheniformis, Pseudomonas aeruginosa, Alcaligenes faecalis, Serratia marcescens, Enterobacter cloacae and Providenicia which were identified as the prominent PAHs-utilizers isolated from CL. Conclusion: This study indicates that the CL samples contain a diverse population of PAHs-degrading bacteria and the use of CL may have a potential for bioremediation of PAHs contaminated sites.

Keywords: polycyclic aromatic hydrocarbons, physico-chemical analyses, compost leachate, microbial and biochemical analyses, phylogenic relations, 16S rDNA sequence analysis

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Authors: Peng Cheng, Liqing Wei, Hongyu Chen, Ruomiao Wang

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This paper discusses a light-weight reinforced thermoplastic (LWRT) composite with superior fire retardancy. This porous LWRT composite is manufactured using polyolefin, fiberglass, and fire retardant additives via a wet-lay process. However, discoloration of the LWRT can be induced by various mechanisms, which may be a concern in the building and construction industry. It is commonly understood that discoloration is strongly associated with the presence of phenolic antioxidant(s) and NO_x. The over-oxidation of phenolic antioxidant(s) is probably the root-cause of the discoloration (pinking/yellowing). Hanwha Azdel, Inc. developed a LWRT with fire-retardant property of ASTM E84-Class A specification, as well as negligible discoloration even under harsh conditions. In addition, this thermoplastic material is suitable for secondary processing (e.g. compression molding) if necessary.

Keywords: discoloration, fire-retardant, thermoplastic composites, wet-lay process

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Authors: Shi-Bo Feng, Sheng-Jie Zhang, Jin Zhou

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The relationship between microbial diversity and algal bloom has received considerable attention for decades. Microbes undoubtedly affect annual bloom events and impact the physiology of both partners, as well as shape ecosystem diversity. However, knowledge about interactions and network correlations among broader-spectrum microbes that lead to the dynamics in a complete bloom cycle are limited. In this study, pyrosequencing and network approaches simultaneously assessed the associate patterns among bacteria, archaea, and microeukaryotes in surface water and sediments in response to a natural dinoflagellate (Alexandrium sp.) bloom. In surface water, among the bacterial community, Gamma-Proteobacteria and Bacteroidetes dominated in the initial bloom stage, while Alpha-Proteobacteria, Cyanobacteria, and Actinobacteria become the most abundant taxa during the post-stage. In the archaea biosphere, it clustered predominantly with Methanogenic members in the early pre-bloom period while the majority of species identified in the later-bloom stage were ammonia-oxidizing archaea and Halobacteriales. In eukaryotes, dinoflagellate (Alexandrium sp.) was dominated in the onset stage, whereas multiply species (such as microzooplankton, diatom, green algae, and rotifera) coexistence in bloom collapse stag. In sediments, the microbial species biomass and richness are much higher than the water body. Only Flavobacteriales and Rhodobacterales showed a slight response to bloom stages. Unlike the bacteria, there are small fluctuations of archaeal and eukaryotic structure in the sediment. The network analyses among the inter-specific associations show that bacteria (Alteromonadaceae, Oceanospirillaceae, Cryomorphaceae, and Piscirickettsiaceae) and some zooplankton (Mediophyceae, Mamiellophyceae, Dictyochophyceae and Trebouxiophyceae) have a stronger impact on the structuring of phytoplankton communities than archaeal effects. The changes in population were also significantly shaped by water temperature and substrate availability (N & P resources). The results suggest that clades are specialized at different time-periods and that the pre-bloom succession was mainly a bottom-up controlled, and late-bloom period was controlled by top-down patterns. Additionally, phytoplankton and prokaryotic communities correlated better with each other, which indicate interactions among microorganisms are critical in controlling plankton dynamics and fates. Our results supplied a wider view (temporal and spatial scales) to understand the microbial ecological responses and their network association during algal blooming. It gives us a potential multidisciplinary explanation for algal-microbe interaction and helps us beyond the traditional view linked to patterns of algal bloom initiation, development, decline, and biogeochemistry.

Keywords: microbial community, harmful algal bloom, ecological process, network

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Authors: R. Gounina-Allouane, I. Moussaoui, N. Boukahel

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Bacillus antimicrobial metabolites, especially those of Bacillus thuringiensis (Bt), are of great interest for research because of health risks generated by the excessive use of chemical additives as well as the propagation of resistant microbial strains, caused by the massive treatment with antibiotics. The objective of this study was the selection of Bt strains producing antimicrobial peptides (bacteriocins), and the partial purification of the most powerful bacteriocins, then the determination of their spectra of antimicrobial action. A collection of twenty one Bt strains isolated from soil at Boumerdès (northern Algeria) was used for screening strains having an antagonistic activity against phylogenetically closed bacteria. Spectra of antagonistic activity of two selected strains was determined against other Bt strains, Gram positive and Gram negative bacterial strains of clinical origin and others from ATCC collection as well as yeasts isolated in human dermatology. Bacteriocins of these two strains were partially purified and their effect on the kinetics of growth of the most sensitive microbial strains was studied. The bacteriocinogenic strains were biochemically characterized and their sensitivity to antibiotics was studied.

Keywords: antimicrobial peptides, Bacillus thuringiensis, bacteriocin, partial purification

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Authors: Mahmoud H. Abu Elella, Riham R. Mohamed, Magdy W. Sabaa

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Introduction: Microbial pollution is global problem threatening the human health. It is resulted by pathogenic microorganisms such as Escherichia coli (E. coli), Staphylococcus aureus (S. aureus) and other pathogenic strains. They cause a dangerous effect on human health, so great efforts have been exerted to produce new and effective antimicrobial agents. Nowadays, natural polysaccharides, such as chitosan and its derivatives are used as antimicrobial agents. The aim of our work is to synthesize of a biodegradable polymer such as N-quaternized chitosan (NQC) then Characterization of NQC by using different analysis techniques such as Fourier transform infrared (FTIR) and Scanning electron microscopy (SEM) and using it as an antibacterial agent against different pathogenic bacteria. Methods: Synthesis of NQC using dimethylsulphate. Results: FTIR technique exhibited absorption peaks of NQC, SEM images illustrated that surface of NQC was smooth and antibacterial results showed that NQC had a high antibacterial effect. Discussion: NQC was prepared and it was proved by FTIR technique and SEM images antibacterial results exhibited that NQC was an antibacterial agent.

Keywords: antimicrobial agent, N-quaternized chitosan chloride, silver nanocomposites, sodium polyacrylate

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Authors: Patrícia Branco, Catarina Prista, Helena Albergaria

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Industrial fuel-ethanol fermentations are carried out under non-sterile conditions, which favors the development of microbial contaminants, leading to huge economic losses. Wild yeasts such as Brettanomyces bruxellensis and lactic acid bacteria are the main contaminants of industrial bioethanol fermentation, affecting Saccharomyces cerevisiae performance and decreasing ethanol yields and productivity. In order to control microbial contaminations, the fuel-ethanol industry uses different treatments, including acid washing and antibiotics. However, these control measures carry environmental risks such as acid toxicity and the rise of antibiotic-resistant bacteria. Therefore, it is crucial to develop and apply less toxic and more environmentally friendly biocontrol methods. In the present study, an industrial fuel-ethanol starter, S. cerevisiae Ethanol-Red, was genetically modified to over-express AMPs with activity against fuel-ethanol microbial contaminants and evaluated regarding its biocontrol effect during mixed-culture alcoholic fermentations artificially contaminated with B. bruxellensis. To achieve this goal, S. cerevisiae Ethanol-Red strain was transformed with a plasmid containing the AMPs-codifying genes, i.e., partial sequences of TDH1 (925-963 bp) and TDH2/3 (925-963 bp) and a geneticin resistance marker. The biocontrol effect of those genetically modified strains was evaluated against B. bruxellensis and compared with the antagonistic effect exerted by the modified strain with an empty plasmid (without the AMPs-codifying genes) and the non-modified strain S. cerevisiae Ethanol-Red. For that purpose, mixed-culture alcoholic fermentations were performed in a synthetic must use the modified S. cerevisiae Ethanol-Red strains together with B. bruxellensis. Single-culture fermentations of B. bruxellensis strains were also performed as a negative control of the antagonistic effect exerted by S. cerevisiae strains. Results clearly showed an improved biocontrol effect of the genetically-modified strains against B. bruxellensis when compared with the modified Ethanol-Red strain with the empty plasmid (without the AMPs-codifying genes) and with the non-modified Ethanol-Red strain. In mixed-culture fermentation with the modified S. cerevisiae strain, B. bruxellensis culturability decreased from 5×104 CFU/mL on day-0 to less than 1 CFU/mL on day-10, while in single-culture B. bruxellensis increased its culturability from 6×104 to 1×106 CFU/mL in the first 6 days and kept this value until day-10. Besides, the modified Ethanol-Red strain exhibited an enhanced antagonistic effect against B. bruxellensis when compared with that induced by the non-modified Ethanol-Red strain. Indeed, culturability loss of B. bruxellensis after 10 days of fermentation with the modified Ethanol-Red strain was 98.7 and 100% higher than that occurred in fermentations performed with the non-modified Ethanol-Red and the empty-plasmid modified strain, respectively. Therefore, one can conclude that the S. cerevisiae genetically modified strain obtained in the present work may be a valuable solution for the mitigation of microbial contamination in fuel-ethanol fermentations, representing a much safer and environmentally friendly preservation strategy than the antimicrobial treatments (acid washing and antibiotics) currently applied in fuel-ethanol industry.

Keywords: antimicrobial peptides, fuel-ethanol microbial contaminations, fuel-ethanol fermentation, biocontrol agents, genetically-modified yeasts

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Authors: Abdulrahman Fahim M. Alsulami

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The current research seeks to explore the regulation of vaccine-related IP rights in light of the areas of divergence between the Trade-Related Aspects of Intellectual Property Rights (TRIPS) Agreement and investment treaties. The study is conducted in the context of the COVID-19 pandemic; therefore, it seems natural that a specific chapter is devoted to the examination of vaccine arrangements related to vaccine supplies. The chapter starts with the examination of a typical vaccine from the perspective of IP rights. It presents the distinctive features of vaccines as pharmaceutical products and investments, reviews the basics of their patent protection, reviews vaccines’ components, and discusses IPR protection of different components of vaccines. The subsection that focuses on vaccine development and licensing reviews vaccine development stages investigates differences between vaccine licensing in different countries and presents barriers to vaccine licensing. The third subsection, at the same time, introduces the existing arrangements related to COVID-19 vaccine supplies, including COVAX arrangements, international organizations’ assistance, and direct negotiations between governments and vaccine manufacturers.

Keywords: bilateral investment treaties, COVID-19 vaccine, IP rights, TRIPs agreement

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Authors: Tatiana A. Kuznetsova, Maxim V. Vechersky, Natalia V. Kostina, Marat M. Umarov, Elena I. Naumova

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One of the main problems of nutrition of phytophagous animals is the insufficiency of protein in their forage. Usually, symbiotic microorganisms highly contribute both to carbohydrates and nitrogen compounds of the food. But it is not easy to utilize microbial biomass in the large intestine and caecum for the animals with hindgut fermentation. So that, some animals, as well hares, developed special mechanism of contribution of such biomass - obligate autocoprophagy, or reingestion. Hares have two types of feces - the hard and the soft. Hard feces are excreted at night, while hares are vigilance ("foraging period"), and the soft ones (caecotrophs) are produced and reingested in the day-time during hares "resting-period". We examine the role of microbial digestion in providing nitrogen nutrition of hare (Lepus europaeus). We determine the ability of nitrogen fixation in fornix and stomach body, small intestine, caecum and colon of hares' gastro-intestinal tract in two main period of hares activity - "resting-period" (day time) and "foraging period" (late-evening and very-early-morning). We use gas chromatography to measure levels of nitrogen fixing activity (acetylene reduction). Nitrogen fixing activity was detected in the contents of all analyzed parts of the gastrointestinal tract. Maximum values were recorded in the large intestine. Also daily dynamics of the process was detected. Thus, during hare “resting-period” (caecotrophs formation) N2-fixing activity was significantly higher than during “foraging period”, reaching 0,3 nmol C2H4/g*h. N2-fixing activity in the gastrointestinal tract can allocate to significant contribution of nitrogen fixers to microbial digestion in hare and confirms the importance of coprophagy as a nitrogen source in lagomorphs.

Keywords: coprophagy, gastrointestinal tract, lagomorphs, nitrogen fixation

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Authors: R. Gounina-Allouane, I. Moussaoui, N. Boukahel

Abstract:

Bacillus antimicrobial metabolites, especially those of Bacillus thuringiensis (Bt), are of great interest for research because of health risks generated by the excessive use of chemical additives as well as the propagation of resistant microbial strains, caused by the massive treatment with antibiotics. The objective of this study was the selection of Bt strains producing antimicrobial peptides (bacteriocins), and the partial purification of the most powerful bacteriocins, then the determination of their spectra of antimicrobial action. A collection of twenty one Bt strains isolated from soil at Boumerdès (northern of Algeria) was used for screening strains having an antagonistic activity against phylogenetically closed bacteria. Spectra of antagonistic activity of two selected strains was determined against other Bt strains, Gram positive and Gram negative bacterial strains of clinical origin and others from ATCC collection as well as yeasts isolated in human dermatology. Bacteriocins of these two strains were partially purified and their effect on the kinetics of growth of the most sensitive microbial strains was studied. The bacteriocinogenic strains were biochemically characterized and their sensitivity to antibiotics was studied.

Keywords: antimicrobial peptides, Bacillus thuringiensis, bacteriocin, partial purification

Procedia PDF Downloads 333

Authors: András Fülöp, Lejla Heilmann, Zsolt Szabó, Ákos Koós

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Microbial fuel cells (MFCs) present a sustainable biotechnological solution to future energy demands. The aim of this study was to construct soil based, single cell, membrane-less MFC systems, operated without treatment to continuously power on-site monitoring and control systems during the soil bioremediation processes. Our Pseudomonas aeruginosa 541 isolate is an ideal choice for MFCs, because it is able to produce pyocyanin which behaves as electron-shuttle molecule, furthermore, it also has a significant antimicrobial effect. We tested several materials and structural configurations to obtain long term high power output. Comparing different configurations, a proton exchange membrane-less, 0.6 m long with 0.05 m diameter MFC tubes offered the best long-term performances. The long-term electricity production were tested from starch, yeast extract (YE), carboxymethyl cellulose (CMC) with humic acid (HA) as a mediator. In all cases, 3 kΩ external load have been used. The two best-operated systems were the Pseudomonas aeruginosa 541 containing MFCs with 1 % carboxymethyl cellulose and the MFCs with 1% yeast extract in the anode area and 35% hydrogel in the cathode chamber. The first had 3.3 ± 0.033 mW/m2 and the second had 4.1 ± 0.065 mW/m2 power density values. These systems have operated for 230 days without any treatment. The addition of 0.2 % HA and 1 % YE referred to the volume of the anode area resulted in 1.4 ± 0.035 mW/m2 power densities. The mixture of 1% starch with 0.2 % HA gave 1.82 ± 0.031 mW/m2. Using CMC as retard carbon source takes effect in the long-term bacterial survivor, thus enable the expression of the long term power output. The application of hydrogels in the cathode chamber significantly increased the performance of the MFC units due to their good water retention capacity.

Keywords: microbial fuel cell, bioremediation, Pseudomonas aeruginosa, biotechnological solution

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Authors: Yongkui Cui, Fengping Wang, Hailei Zhao, Muhammad Zubair Iqbal, Ziya Wang, Yan Li, Pengpeng LV

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The novel 3D SnO cabbages self-assembled by nanosheets were successfully synthesized via template-free hydrothermal growth method under facile conditions.The XRD results manifest that the as-prepared SnO is tetragonal phase. The TEM and HRTEM results show that the cabbage nanosheets are polycrystalline structure consisted of considerable single-crystalline nanoparticles. Two typical Raman modes A1g=210 and Eg=112 cm-1 of SnO are observed by Raman spectroscopy. Moreover, galvanostatic cycling tests has been performed using the SnO cabbages as anode material of lithium ion battery and the electrochemical results suggest that the synthesized SnO cabbage structures are a promising anode material for lithium ion batteries.

Keywords: electrochemical property, hydrothermal synthesis, lithium ion battery, stannous oxide

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Authors: Islem Kedidi, Rihab Bedoui Bensalem, Faysal Manssouri

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In traditional models of insurance data, the number and size of claims are assumed to be independent. Relaxing the independence assumption, this article explores the Vine copula to model dependence structure between multivariate frequency and average severity of insurance claim. To illustrate this approach, we use the Wisconsin local government property insurance fund which offers several insurance protections for motor vehicles, property and contractor’s equipment claims. Results show that the C-vine copula can better characterize the multivariate dependence structure between frequency and severity. Furthermore, we find significant dependencies especially between frequency and average severity among different coverage types.

Keywords: dependency modeling, government insurance, insurance claims, vine copula

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Authors: N. Mezıou-Cheboutı, A. Merabet, N. Behidj, F. Z. Bissaad

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The Algeria by its location, offers a rich and diverse vegetation. A large number of aromatic and medicinal plants grow spontaneously. The interest in these plants has continued to grow in recent years. Their particular properties due to the essential oil fraction can be utilized to treat microbial infections. To this end, and in the context of the valuation of the Algerian flora, we became interested in the species of the family Lamiaceae which is one of the most used as a global source of spices and extracts strong families antimicrobial potency. The plant on which we have based our choice is a species of sage "Salvia officinalis" from the Isser localized region within the province of Boumerdes. This work focuses on the study of the antimicrobial activity of essential oil extracted from the leaves of salvia officinalis. The extraction is carried out by HE hydrodistillation and reveals a yield of 1.06℅. The study of the antimicrobial activity of the essential oil by the method of at aromatogramme shown that Gram positive bacteria are most susceptible (Staphylococcus aureus and Bacillus subtilis) with a strong inhibition of growth. The yeast Candida albicans fungus Aspergillus niger and have shown moderately sensitive.

Keywords: Salvia officinalis, steam distillation, essential oil, aromatogram, anti-microbial activity

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Authors: Akshita Bhardwaj

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Keratinase is an enzyme obtained from extracellular sources that is involved in biodegradation of keratin. It is a member of a group of proteases that can break down keratin into amino acids. Keratinases are produced only in the presence of substrate that contain keratin. It attacked the disulfide bond of substrate and involve in keratin degradation. Human hair, feathers, animal hard tissues, horns, claws, and hooves all contain keratin.. It exists in two form alpha keratin (found in soft tissues) and beta keratin (found in hard tissue). By taking part in the degradation of keratin, keratinases derived from microbial sources, often referred to as microbial keratinases, are important in the process of turning wastes containing keratin into products with added value. Chicken feathers contain high level of keratin protein content than other sources and became a suitable protein source. Keratinase production occurs at near alkaline pH and thermophilic temperatures. The bioprocessing of keratinous waste benefits greatly from the use of keratinases. Additionally, it lessens the issue caused by poultry excrement. The use of feather meal, along with keratinase, improves the digestion of proteins and amino acids.

Keywords: mili litre (ml), micro litre (Ul), TCA - trichloroacetic acid, OD - optical density

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Authors: Hakima Althalb

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Petroleum contamination of sandy soils is a severe environmental problem in Libya, but relatively little work has been carried out to optimize the bioremediation of such heavily contaminated soil, particularly at a pilot scale. The purpose of this research was to determine the potential for the microbial-based bioremediation of hydrocarbon-contaminated soil obtained from an oil refinery in Libya and to assess the potential of both ozonation and phytoremediation (both applied after initial bioremediation) to reduce residual hydrocarbon levels. Plots containing 500 kg soil (triplicates) (contaminated soil diluted with clean soil 50% volume) were set up, (designated as Land Treatment Units; LTUs) containing five different nutrient levels and mixtures (Urea + NPK (nitrogen; phosphor; potassium) mixtures) to obtain C:N:P ratios 100:10:1, and monitored for 90 days. Hydrocarbon levels, microbial numbers, and toxicity (EC50 using luminescent microbial based tests) were assessed. Hydrocarbon levels in non-diluted and diluted soil ranged from 20 733-22 366 mg/kg and from 16 000-17 000 mg/kg respectively. Although all the land treatment units revealed a significant hydrocarbon reduction over time, the highest reduction in hydrocarbon levels obtained was around 60%. For example, 63% hydrocarbon removal was observed using a mixture of urea and NPK with a C:N:P ratio of 100:10:1). Soil toxicity (as assessed using luminescence based toxicity assays) reduced in line with the reduction in total petroleum hydrocarbons observed. However, as relatively high residual TPH (total petroleum hydrocarbon) levels (ranging from 6033-14166mg/kg) were still present after initial bioremediation two ‘post-treatments’ (phytoremediation and ozonation) were attempted to remove residual hydrocarbons remaining. Five locally grown (agriculturally important) plant species were tested. The germination of all plants examined was strongly inhibited (80-100%) and seedlings failed to grow well in the contaminated soil, indicating that the previously bioremediated soils were still toxic to the plants. Subsequent ozonation followed by another bioremediation of soil was more successful than phytoremediation. But even the most promising successful treatment in this study (ozonation for 6 hours at 25ppm followed by bioremediation) still only removed approximately 31% of the residual hydrocarbons. Overall, this work showed that the bioremediation of such highly contaminated soils is difficult and that a combination of treatments would be required to achieve successful remediation. Even after initial dilution and bioremediation the soils remained toxic to plant growth and were therefore not suitable for phytoremediation.

Keywords: bioremediation, petroleum hydrocarbons, ozone, phytoremediation

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Authors: Alisa Kazarina, Guntis Gerhards, Elina Petersone-Gordina, Ilva Pole, Viktorija Igumnova, Janis Kimsis, Valentina Capligina, Renate Ranka

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Human body is inhabited by a vast number of microorganisms, collectively known as the human microbiome, and there is a tremendous interest in evolutionary changes in human microbial ecology, diversity and function. The field of paleomicrobiology, study of ancient human microbiome, is powered by modern techniques of Next Generation Sequencing (NGS), which allows extracting microbial genomic data directly from archaeological sample of interest. One of the major techniques is 16S rRNA gene sequencing, by which certain 16S rRNA gene hypervariable regions are being amplified and sequenced. However, some limitations of this method exist including the taxonomic precision and efficacy of different regions used. The aim of this study was to evaluate the phylogenetic sensitivity of different 16S rRNA gene hypervariable regions for microbiome studies in the archaeological samples. Towards this aim, archaeological bone samples and corresponding soil samples from each burial environment were collected in Medieval cemeteries in Latvia. The Ion 16S™ Metagenomics Kit targeting different 16S rRNA gene hypervariable regions was used for library construction (Ion Torrent technologies). Sequenced data were analysed by using appropriate bioinformatic techniques; alignment and taxonomic representation was done using Mothur program. Sequences of most abundant genus were further aligned to E. coli 16S rRNA gene reference sequence using MEGA7 in order to identify the hypervariable region of the segment of interest. Our results showed that different hypervariable regions had different discriminatory power depending on the groups of microbes, as well as the nature of samples. On the basis of our results, we suggest that wider range of primers used can provide more accurate recapitulation of microbial communities in archaeological samples. Acknowledgements. This work was supported by the ERAF grant Nr. 1.1.1.1/16/A/101.

Keywords: 16S rRNA gene, ancient human microbiome, archaeology, bioinformatics, genomics, microbiome, molecular biology, next-generation sequencing

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Authors: Claudivan Costa de Lima, Angelo da Silva Monteiro

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With the aim of evaluating the effects of grounding with copper wire on the reduction of the microbial load of biocontaminated milk samples and on their acidification over time, two complementary experiments were carried out. In the first, the treatments consisted of: i) raw milk sample (control), ii) slow pasteurization, iii) grounding with copper wire and, iv) contact with copper ring. Analyzes of total, thermoresistant and mesophilic coliforms were performed 30 minutes after the application of these treatments. In the second experiment, under the same conditions as the first, measurements of pH and Dornic acidity were performed at 0, 0.5, 2, 4, 8, 12, and 24 h from the installation of the experiment. Pasteurization eliminated almost all groups of bacteria present in the milk samples while grounding only allowed reductions in the population of thermotolerant coliforms and mesophiles, both greater than 95%, maintaining, however, unchanged the amounts of total coliforms. The copper ring, in turn, had no effect on the microbiological parameters studied. The reduction in the population of mesophiles in grounded milk samples, contrary to what happened with pasteurized milk, was not enough to inhibit the acidification process over the experimental period.

Keywords: pasteurization, low frequency electric current, thermotolerant coliforms, mesophiles in bovine milk

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Authors: N. Balaban, A. Buernstein, F. Gelman, Z. Ronen

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Brominated flame retardants are widespread pollutants, and are known to be toxic, carcinogenic, endocrinic disrupting as well as recalcitrant. The industrial complex Neot Hovav, in the Northern Negev, Israel, is situated above a fractured chalk aquitard, which is polluted by a wide variety of halogenated organic compounds. Two of the abundant pollutants found in the site are Dibromoneopentyl-glycol (DBNPG) and tribromoneopentyl-alcohol (TBNPA). Due to the elusive nature of the groundwater flow, it is difficult to connect between the spatial changes in contaminant concentrations to degradation. In this study, we attempt to determine whether these compounds are biodegraded in the groundwater, and to gain a better understanding concerning the bacterial community in the groundwater. This was achieved through the application of compound-specific isotope analysis (CSIA) of carbon (13^C/12^C) and bromine (81^Br/79^Br), and new-generation MiSeq pyrosequencing. The sampled boreholes were distributed among three main areas of the industrial complex: around the production plant of TBNPA and DBNPG; along the Hovav Wadi (small ephemeral stream) which crosses and drains the industrial complex; and downstream to the industrial area. TBNPA and DBNPG are found in all three areas, with no clear connection to the proximity of the borehole to the production plant. Initial isotopic data of TBNPA from boreholes in the area surrounding the production plant, reveal no changes in the carbon and bromine isotopic values. When observing the microbial groundwater community, the dominant phylum is Proteobacteria. Known anaerobic dehalogenating bacteria such as Dehalococcoides from the Chloroflexi phylum have also been detected. A statistical comparison of the groundwater microbial diversity using a multi-variant ordination of non-metric multidimensional scaling (NMDS) reveals three main clusters in accordance to spatial location in the industrial complex: all the boreholes sampled adjacent to the production plant cluster together and separately from the Wadi Hovav boreholes cluster and the downstream to the industrial area borehole cluster. This work provides the basis for the development and implication of an isotopic fractionation based tool for assessing the biodegradation of brominated organic compounds in contaminated environments, and a novel attempt to characterize the spatial microbial diversity in the contaminated site.

Keywords: biodegradation, brominated flame retardants, groundwater, isotopic fractionation, microbial diversity

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Authors: Dheepshika Kodieswaran

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The persisting problem in the world that continuously impose our planet at risk is the increasing amounts of recalcitrant. One such issue is the disposal of the Petroleum Refinery Sludge (PRS) which constitutes hydrocarbons that are hazardous to terrestrial and aquatic life. The comparatively safe approach to handling these wastes is by microbial degradation, while the other chemical and physical methods are either expensive and/or produce secondary pollutants. The bacterial and algal systems have different pathways for the degradation of hydrocarbons, and their growth rates vary. This study shows how different bacterial and microalgal strains degrade the polyaromatic hydrocarbon PAHs individually and their symbiotic influence on degradation as well. In this system, the metabolites and gaseous exchange help each other in growth. This method using also aids in the accumulation of lipids in microalgal cells and from which bio-oils can also be extracted. The bacterial strains used in this experiment are reported to be indigenous strains isolated from PRS. The target PAH studied were anthracene and pyrene for a period of 28 days. The PAH degradation kinetics best fitted the Gompertz model, and the order of the kinetics, rate constants, and half-life was determined.

Keywords: petroleum refinery sludge, co-culturing, polycyclic hydrocarbons, microalgal-bacterial consortia

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Authors: Isabel N. Sierra-Garcia, Maria J. Ferreira, Sandro Figuereido, Newton Gomes, Helena Silva, Angela Cunha

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Plant-associated microbial communities are considered crucial in the adaptation of halophytes to coastal environments. The plant microbiota can be horizontally acquired from the environment or vertically transmitted from generation to generation via seeds. Recruiting of the microbial communities by the plant is affected by geographical location, soil source, host genotype, and cultivation practice. There is limited knowledge reported on the microbial communities in halophytes the influence of biotic and abiotic factors. In this work, the microbiota associated with the halophyte Salicornia ramosissima was investigated to determine whether the structure of bacterial communities is influenced by host genotype or soil source. For this purpose, two contrasting sites where S. ramosissima is established in the estuarine system of the Ria de Aveiro were investigated. One site corresponds to a natural salt marsh where S. ramosissima plants are present (wild plants), and the other site is a former salt pan that nowadays are subjected to intensive crop production of S. ramosissima (crop plants). Bacterial communities from the rhizosphere, seeds and root endosphere of S. ramossisima from both sites were investigated by sequencing bacterial 16S rRNA gene using the Illumina MiSeq platform. The analysis of the sequences showed that the three plant-associated compartments, rhizosphere, root endosphere, and seed endosphere, harbor distinct microbiomes. However, bacterial richness and diversity were higher in seeds of wild plants, followed by rhizosphere in both sites, while seeds in the crop site had the lowest diversity. Beta diversity measures indicated that bacterial communities in root endosphere and seeds were more similar in both wild and crop plants in contrast to rhizospheres that differed by local, indicating that the recruitment of the similar bacterial communities by the plant genotype is active in regard to the site. Moreover, bacterial communities from the root endosphere and rhizosphere were phylogenetically more similar in both sites, but the phylogenetic composition of seeds in wild and crop sites was distinct. These results indicate that cultivation practices affect the seed microbiome. However, minimal vertical transmission of bacteria from seeds to adult plants is expected. Seeds from the crop site showed higher abundances of Kushneria and Zunongwangia genera. Bacterial members of the classes Alphaprotebacteria and Bacteroidia were the most ubiquitous across sites and compartments and might encompass members of the core microbiome. These findings indicate that bacterial communities associated with S. ramosissima are more influenced by host genotype rather than local abiotic factors or cultivation practices. This study provides a better understanding of the composition of the plant microbiota in S. ramosissima , which is essential to predict the interactions between plant and associated microbial communities and their effects on plant health. This knowledge is useful to the manipulations of these microbial communities to enhance the health and productivity of this commercially important plant.

Keywords: halophytes, plant microbiome, Salicornia ramosissima, agriculture

Procedia PDF Downloads 132