Search results for: bacteria and Fungi.
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 310

Search results for: bacteria and Fungi.

190 Biodegradation of Cyanide by a Novel Cyanidedegrading Bacterium

Authors: S. Potivichayanon, R. Kitleartpornpairoat

Abstract:

The objectives were to identify cyanide-degrading bacteria and study cyanide removal efficiency. Agrobacterium tumefaciens SUTS 1 was isolated. This is a new strain of microorganisms for cyanide degradation. The maximum growth rate of SUTS 1 obtained 4.7 × 108 CFU/ml within 4 days. The cyanide removal efficiency was studied at 25, 50, and 150 mg/L cyanide. The residual cyanide, ammonia, nitrate, nitrite, pH, and cell counts were analyzed. At 25 and 50 mg/L cyanide, SUTS 1 obtained similar removal efficiency approximately 87.50%. At 150 mg/L cyanide, SUTS 1 enhanced the cyanide removal efficiency up to 97.90%. Cell counts of SUTS 1 increased when the cyanide concentration was set at lower. The ammonia increased when the removal efficiency increased. The nitrate increased when the ammonia decreased but the nitrite did not detect in all experiments. pH values also increased when the cyanide concentrations were set at higher.

Keywords: Biodegradation, Cyanide-degrading bacteria, Removal efficiency, Residual cyanide

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189 A Green Chemical Technique for the Synthesis of Magnetic Nanoparticles by Magnetotactic Bacteria

Authors: Parisa Tajer-Mohammad-Ghazvini, Rouha Kasra-Kermanshahi, Ahmad Nozad-Golikand, Majid Sadeghizadeh

Abstract:

Bacterial magnetic nanoparticles have great useful potential in biotechnological and biomedical applications. In this study, a liquid growth medium was modified for cultivation a fastidious magnetotactic bacterium that has been isolated from Anzali lagoon, Iran in our previous research. These modifications include change in vitamin, mineral, carbon sources and etcetera. In our experience, the serum bottles and designed air-tight laboratory bottles were used to create microaerobic conditions in order to development of a method for scale-up experiment. This information may serve as a guide to green chemistry based biological protocols for the synthesis of magnetic nanoparticles with control over the chemical composition, morphology and size.

Keywords: Green chemistry, Magnetosome, Magnetotactic bacteria, Magnetic nanoparticles, Nano-Biotechnology.

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188 Biosynthesis of Silver-Phosphate Nanoparticles Using the Extracellular Polymeric Substance of Sporosarcina pasteurii

Authors: Mohammadhosein Rahimi, Mohammad Raouf Hosseini, Mehran Bakhshi, Alireza Baghbanan

Abstract:

Silver ions (Ag+) and their compounds are consequentially toxic to microorganisms, showing biocidal effects on many species of bacteria. Silver-phosphate (or silver orthophosphate) is one of these compounds, which is famous for its antimicrobial effect and catalysis application. In the present study, a green method was presented to synthesis silver-phosphate nanoparticles using Sporosarcina pasteurii. The composition of the biosynthesized nanoparticles was identified as Ag3PO4 using X-ray Diffraction (XRD) and Energy Dispersive Spectroscopy (EDS). Also, Fourier Transform Infrared (FTIR) spectroscopy showed that Ag3PO4 nanoparticles was synthesized in the presence of biosurfactants, enzymes, and proteins. In addition, UV-Vis adsorption of the produced colloidal suspension approved the results of XRD and FTIR analyses. Finally, Transmission Electron Microscope (TEM) images indicated that the size of the nanoparticles was about 20 nm.

Keywords: Bacteria, biosynthesis, silver-phosphate, Sporosarcina pasteurii, nanoparticle.

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187 Screening of Antagonistic/Synergistic Effect between Lactic Acid Bacteria (LAB) and Yeast Strains Isolated from Kefir

Authors: Mihriban Korukluoglu, Goksen Arik, Cagla Erdogan, Selen Kocakoglu

Abstract:

Kefir is a traditional fermented refreshing beverage which is known for its valuable and beneficial properties for human health. Mainly yeast species, lactic acid bacteria (LAB) strains and fewer acetic acid bacteria strains live together in a natural matrix named “kefir grain”, which is formed from various proteins and polysaccharides. Different microbial species live together in slimy kefir grain and it has been thought that synergetic effect could take place between microorganisms, which belong to different genera and species. In this research, yeast and LAB were isolated from kefir samples obtained from Uludag University Food Engineering Department. The cell morphology of isolates was screened by microscopic examination. Gram reactions of bacteria isolates were determined by Gram staining method, and as well catalase activity was examined. After observing the microscopic/morphological and physical, enzymatic properties of all isolates, they were divided into the groups as LAB and/or yeast according to their physicochemical responses to the applied examinations. As part of this research, the antagonistic/synergistic efficacy of the identified five LAB and five yeast strains to each other were determined individually by disk diffusion method. The antagonistic or synergistic effect is one of the most important properties in a co-culture system that different microorganisms are living together. The synergistic effect should be promoted, whereas the antagonistic effect is prevented to provide effective culture for fermentation of kefir. The aim of this study was to determine microbial interactions between identified yeast and LAB strains, and whether their effect is antagonistic or synergistic. Thus, if there is a strain which inhibits or retards the growth of other strains found in Kefir microflora, this circumstance shows the presence of antagonistic effect in the medium. Such negative influence should be prevented, whereas the microorganisms which have synergistic effect on each other should be promoted by combining them in kefir grain. Standardisation is the most desired property for industrial production. Each microorganism found in the microbial flora of a kefir grain should be identified individually. The members of the microbial community found in the glue-like kefir grain may be redesigned as a starter culture regarding efficacy of each microorganism to another in kefir processing. The main aim of this research was to shed light on more effective production of kefir grain and to contribute a standardisation of kefir processing in the food industry.

Keywords: Antagonistic effect, kefir, lactic acid bacteria (LAB), synergistic, yeast.

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186 Microbial Contaminants in Drinking Water Collected from Different Regions of Kuwait

Authors: Abu Salim Mustafa

Abstract:

Water plays a major role in maintaining life on earth, but it can also serve as a matrix for pathogenic organisms, posing substantial health threats to humans. Although, outbreaks of diseases attributable to drinking water may not be common in industrialized countries, they still occur and can lead to serious acute, chronic, or sometimes fatal health consequences. The analysis of drinking water samples from different regions of Kuwait was performed in this study for bacterial and viral contaminations. Drinking tap water samples were collected from 15 different locations of the six Kuwait governorates. All samples were analyzed by confocal microscopy for the presence of bacteria. The samples were cultured in vitro to detect cultivable organisms. DNA was isolated from the cultured organisms and the identity of the bacteria was determined by sequencing the bacterial 16S rRNA genes, followed by BLAST analysis in the database of NCBI, USA. RNA was extracted from water samples and analyzed by real-time PCR for the detection of viruses with potential health risks, i.e. Astrovirus, Enterovirus, Norovirus, Rotavirus, and Hepatitis A. Confocal microscopy showed the presence of bacteria in some water samples. The 16S rRNA gene sequencing of culture grown organisms, followed by BLAST analysis, identified the presence of several non-pathogenic bacterial species. However, one sample had Acinetobacter baumannii, which often causes opportunistic infections in immunocompromised people, but none of the studied viruses could be detected in the drinking water samples analyzed. The results indicate that drinking water samples analyzed from various locations in Kuwait are relatively safe for drinking and do not contain many harmful pathogens.

Keywords: Drinking water, 16S rRNA, microbial diversity, viruses, Kuwait.

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185 An in Silico Approach for Prioritizing Drug Targets in Metabolic Pathway of Mycobacterium Tuberculosis

Authors: Baharak Khoshkholgh-Sima, Soroush Sardari, Jalal Izadi Mobarakeh, Ramezan Ali Khavari-Nejad

Abstract:

There is an urgent need to develop novel Mycobacterium tuberculosis (Mtb) drugs that are active against drug resistant bacteria but, more importantly, kill persistent bacteria. Our study structured based on integrated analysis of metabolic pathways, small molecule screening and similarity Search in PubChem Database. Metabolic analysis approaches based on Unified weighted used for potent target selection. Our results suggest that pantothenate synthetase (panC) and and 3-methyl-2-oxobutanoate hydroxymethyl transferase (panB) as a appropriate drug targets. In our study, we used pantothenate synthetase because of existence inhibitors. We have reported the discovery of new antitubercular compounds through ligand based approaches using computational tools.

Keywords: In Silico, Ligand-based Virtual Screening, Metabolic Pathways, Mycobacterium tuberculosis

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184 Bioleaching for Efficient Copper Ore Recovery

Authors: Zh. Karaulova, D. Baizhigitov

Abstract:

At the Aktogay deposit, the oxidized ore section has been developed since 2015; by now, the reserves of easily enriched ore are decreasing, and a large number of copper-poor, difficult-to-enrich ores has been accumulated in the dumps of the KAZ Minerals Aktogay deposit, which is unprofitable to mine using the traditional mining methods. Hence, another technology needs to be implemented, which will significantly expand the raw material base of copper production in Kazakhstan and ensure the efficient use of natural resources. Heap and dump bacterial recovery are the most acceptable technologies for processing low-grade secondary copper sulfide ores. Test objects were the copper ores of Aktogay deposit and chemolithotrophic bacteria Leptospirillum ferrooxidans (L.f.), Acidithiobacillus caldus (A.c.), Sulfobacillus acidophilus (S.a.), represent mixed cultures utilized in bacterial oxidation systems. They can stay active in the 20-40 °C temperature range. Biocatalytic acceleration was achieved as a result of bacteria oxidizing iron sulfides to form iron sulfate, which subsequently underwent chemical oxidation to become sulfate oxide. The following results have been achieved at the initial stage: the goal was to grow and maintain the life activity of bacterial cultures under laboratory conditions. These bacteria grew the best within the pH 1,2-1,8 range with light stirring and in an aerated environment. The optimal growth temperature was 30-33 оC. The growth rate decreased by one-half for each 4-5 °C fall in temperature from 30 °C. At best, the number of bacteria doubled every 24 hours. Typically, the maximum concentration of cells that can be grown in ferrous solution is about 107/ml. A further step researched in this case was the adaptation of microorganisms to the environment of certain metals. This was followed by mass production of inoculum and maintenance for their further cultivation on a factory scale. This was done by adding sulfide concentrate, allowing the bacteria to convert the ferrous sulfate as indicated by the Eh (> 600 mV), then diluting to double the volume and adding concentrate to achieve the same metal level. This process was repeated until the desired metal level and volumes were achieved. The final stage of bacterial recovery was the transportation and irrigation of secondary sulfide copper ores of the oxidized ore section. In conclusion, the project was implemented at the Aktogay mine since the bioleaching process was prolonged. Besides, the method of bacterial recovery might compete well with existing non-biological methods of extraction of metals from ores.

Keywords: Bacterial recovery, copper ore, bioleaching, bacterial inoculum.

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183 Acute and Chronic Effect of Biopesticide on Infestation of Whitefly Bemisia tabaci (Gennadius) on the Culantro Cultivation

Authors: U. Pangnakorn, S. Chuenchooklin

Abstract:

Acute and chronic effects of biopesticide from entomopathogenic nematode (Steinernema thailandensis n. sp.), bacteria ISR (Pseudomonas fluorescens), wood vinegar and fermented organic substances from plants: (neem Azadirachta indica + citronella grass Cymbopogon nardus Rendle + bitter bush Chromolaena odorata L.) were tested on culantro (Eryngium foetidum L.). The biopesticide was investigated for infestation reduction of the major insect pest whitefly (Bemisia tabaci (Gennadius)). The experimental plots were located at a farm in Nakhon Sawan Province, Thailand. This study was undertaken during the drought season (late November to May). Effectiveness of the treatment was evaluated in terms of acute and chronic effect. The populations of whitefly were observed and recorded every hour up to 3 hours with insect nets and yellow sticky traps after the treatments were applied for the acute effect. The results showed that bacteria ISR had the highest effectiveness for controlling whitefly infestation on culantro; the whitefly numbers on insect nets were 12.5, 10.0 and 7.5 after 1 hr, 2 hr, and 3 hr, respectively while the whitefly on yellow sticky traps showed 15.0, 10.0 and 10.0 after 1 hr, 2 hr, and 3 hr, respectively. For chronic effect, the whitefly was continuously collected and recorded at weekly intervals; the result showed that treatment of bacteria ISR found the average whitefly numbers only 8.06 and 11.0 on insect nets and sticky traps respectively, followed by treatment of nematode where the average whitefly was 9.87 and 11.43 on the insect nets and sticky traps, respectively. In addition, the minor insect pests were also observed and collected. The biopesticide influenced the reduction number of minor insect pests (red spider mites, beet armyworm, short-horned grasshopper, pygmy locusts, etc.) with only a few found on the culantro cultivation.

Keywords: Whitefly (Bemisia tabaci Gennadius), Culantro (Eryngium foetidum L.), Entomopathogenic nematode (Steinernema thailandensis n. sp.), Bacteria ISR (Pseudomonas fluorescens), wood vinegar, fermented organic substances.

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182 Chemical Characterization and Prebiotic Effect of Water-Soluble Polysaccharides from Zizyphus lotus Leaves

Authors: Zakaria Boual, Abdellah Kemassi, Toufik Chouana, Philippe Michaud, Mohammed Didi Ould El Hadj

Abstract:

In order to investigate the prebiotic potential of oligosaccharides prepared by chemical hydrolysis of water-soluble polysaccharides (WSP) from Zizyphus lotus leaves, the effect of oligosaccharides on bacterial growth was studied. The chemical composition of WSP was evaluated by colorimetric assays revealed the average values: 7.05±0.73% proteins and 86.21±0.74% carbohydrates, among them 64.81±0.42% is neutral sugar and the rest 16.25±1.62% is uronic acids. The characterization of monosaccharides was determined by high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) was found to be composed of galactose (23.95%), glucose (21.30%), rhamnose (20.28%), arabinose (9.55%), and glucuronic acid (22.95%). The effects of oligosaccharides on the growth of lactic acid bacteria were compared with those of fructooligosaccharide (RP95). The oligosaccharides concentration was 1g/L of Man, Rogosa, Sharpe broth. Bacterial growth was assessed during 2, 4.5, 6.5, 9, 12, 16 and 24 h by measuring the optical density of the cultures at 600 nm (OD600) and pH values. During fermentation, pH in broth cultures decreased from 6.7 to 5.87±0.15. The enumeration of lactic acid bacteria indicated that oligosaccharides led to a significant increase in bacteria (P≤0.05) compared to the control. The fermentative metabolism appeared to be faster on RP95 than on oligosaccharides from Zizyphus lotus leaves. Both RP95 and oligosaccharides showed clear prebiotic effects, but had differences in fermentation kinetics because of to the different degree of polymerization. This study shows the prebiotic effectiveness of oligosaccharides, and provides proof for the selection of leaves of Zizyphus lotus for use as functional food ingredients.

Keywords: Zizyphus lotus, polysaccharides, characterization, prebiotic effects.

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181 The Use of Bituminaria bituminosa (L.) Stirton and Microbial Biotechnologies for Restoration of Degraded Pastoral Lands: The Case of the Middle Atlas of Morocco

Authors: O. Zennouhi, M. El Mderssa, J. Ibijbijen, E. Bouiamrine, L. Nassiri

Abstract:

Rangelands and silvopastoral systems of the middle Atlas are under a heavy pressure, which led to pasture degradation, invasion by non-palatable and toxic species and edaphic aridification due to the regression of the global vegetation cover. In this situation, the introduction of multipurpose leguminous shrubs, such as Bituminaria bituminosa (L.) Stirton, commonly known as bituminous clover, could be a promising socio-ecological alternative for the rehabilitation of these degraded areas. The application of biofertilizers like plant growth promoting rhizobacteria especially phosphate solubilizing bacteria (PSB) can ensure a successful installation of this plant in the selected degraded areas. The main objective of the present work is to produce well-inoculated seedlings using the best efficient PSB strains in the greenhouse to increase their ability to resist to environmental constraints once transplanted to the field in the central Middle Atlas.

Keywords: Biofertilizers, Bituminaria bituminosa, phosphate solubilizing bacteria, rehabilitation.

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180 A Maximum Parsimony Model to Reconstruct Phylogenetic Network in Honey Bee Evolution

Authors: Usha Chouhan, K. R. Pardasani

Abstract:

Phylogenies ; The evolutionary histories of groups of species are one of the most widely used tools throughout the life sciences, as well as objects of research with in systematic, evolutionary biology. In every phylogenetic analysis reconstruction produces trees. These trees represent the evolutionary histories of many groups of organisms, bacteria due to horizontal gene transfer and plants due to process of hybridization. The process of gene transfer in bacteria and hybridization in plants lead to reticulate networks, therefore, the methods of constructing trees fail in constructing reticulate networks. In this paper a model has been employed to reconstruct phylogenetic network in honey bee. This network represents reticulate evolution in honey bee. The maximum parsimony approach has been used to obtain this reticulate network.

Keywords: Hybridization, HGT, Reticulate networks, Recombination, Species, Parsimony.

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179 Antimicrobial Activity of Girardinia heterophylla

Authors: P. S. Bedi, Neayti Thakur, Balvinder Singh

Abstract:

In the present study an attempt has been made to prepare the crude extracts of leaves and stem of ‘Girardinia heterophylla’ by using various solvents like petroleum ether, ethanol and double distilled water. The samples were given the code NGLS 1, NGLS 2, NGLS 3 and NGSS 1, NGSS 2 and NGSS 3 respectively. All the extracts were used to study their antimicrobial activity against gram positive bacteria e.g. Bacillus subtilis, gram negative bacteria e.g. E. coli and K. pneumonia and antifungal activity against Aspergillus niger. The results of the antimicrobial activity showed that all the crude extracts of the plant possesses antibacterial activity. Maximum antibacterial activity was shown by NGLS 2, NGLS 3 and NGSS 3 against K. pneumonia. The growth of fungus A. niger was also inhibited by all the crude extracts. Maximum inhibition was shown by NGSS 2 followed by NGSS 1.

Keywords: Girardinia heterophylla, leaves and stem extracts, antibacterial activity, antifungal activity.

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178 Novel Structural Insights of Glutamate Racemase from Mycobacterium tuberculosis through Modeling and Docking Studies

Authors: Jayashree Ramana

Abstract:

An alarming emergence of multidrug-resistant strains of the tuberculosis pathogen Mycobacterium tuberculosis and continuing high worldwide incidence of tuberculosis has invigorated the search for novel drug targets. The enzyme glutamate racemase (MurI) in bacteria catalyzes the stereoconversion of L-glutamate to D-glutamate which is a component of the peptidoglycan cell wall of the bacterium. The inhibitors targeted against MurI from several bacterial species have been patented and are advocated as promising antibacterial agents. However there are none available against MurI from Mycobacterium tuberculosis, due to the lack of its threedimensional structure. This work accomplished two major objectives. First, the tertiary structure of MtMurI was deduced computationally through homology modeling using the templates from bacterial homologues. It is speculated that like in other Gram-positive bacteria, MtMurI exists as a dimer and many of the protein interactions at the dimer interface are also conserved. Second, potent candidate inhibitors against MtMurI were identified through docking against already known inhibitors in other organisms.

Keywords: Glutamate racemase, homology modeling, docking, drug resistance.

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177 Starch Based Biofilms for Green Packaging

Authors: Roshafima R. Ali, W. A. Wan Abdul Rahman, Rafiziana M. Kasmani, N. Ibrahim

Abstract:

This current research focused on development of degradable starch based packaging film with enhanced mechanical properties. A series of low density polyethylene (LDPE)/tapioca starch compounds with various tapioca starch contents were prepared by twin screw extrusion with the addition of maleic anhydride grafted polyethylene as compatibilizer. Palm cooking oil was used as processing aid to ease the blown film process, thus, degradable film can be processed via conventional blown film machine. Studies on their characteristics, mechanical properties and biodegradation were carried out by Fourier Transform Infrared (FTIR) spectroscopy and optical properties, tensile test and exposure to fungi environment respectively. The presence of high starch contents had an adverse effect on the tensile properties of LDPE/tapioca starch blends. However, the addition of compatibilizer to the blends improved the interfacial adhesion between the two materials, hence, improved the tensile properties of the films. High content of starch amount also was found to increase the rate of biodegradability of LDPE/tapioca starch films. It can be proved by exposure of the film to fungi environment. A growth of microbes colony can be seen on the surface of LDPE/tapioca starch film indicates that the granular starch present on the surface of the polymer film is attacked by microorganisms, until most of it is assimilated as a carbon source.

Keywords: Degradable polymer, starch based biofilms, blown film extrusion, green food packaging.

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176 Microbiological Analysis, Cytotoxic and Genotoxic Effects from Material Captured in PM2.5 and PM10 Filters Used in the Aburrá Valley Air Quality Monitoring Network (Colombia)

Authors: Carmen E. Zapata, Juan Bautista, Olga Montoya, Claudia Moreno, Marisol Suarez, Alejandra Betancur, Duvan Nanclares, Natalia A. Cano

Abstract:

This study aims to evaluate the diversity of microorganisms in filters PM2.5 and PM10; and determine the genotoxic and cytotoxic activity of the complex mixture present in PM2.5 filters used in the Aburrá Valley Air Quality Monitoring Network (Colombia). The research results indicate that particulate matter PM2.5 of different monitoring stations are bacteria; however, this study of detection of bacteria and their phylogenetic relationship is not complete evidence to connect the microorganisms with pathogenic or degrading activities of compounds present in the air. Additionally, it was demonstrated the damage induced by the particulate material in the cell membrane, lysosomal and endosomal membrane and in the mitochondrial metabolism; this damage was independent of the PM2.5 concentrations in almost all the cases.

Keywords: Cytotoxic, genotoxic, microbiological analysis, PM10, PM2.5.

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175 Evaluation of Bacterial Composition of the Aerosol of Selected Abattoirs in Akure, South Western Nigeria

Authors: Funmilola O. Omoya, Joseph O. Obameso, Titus A. Olukibiti

Abstract:

This study was carried out to reveal the bacterial composition of aerosol in the studied abattoirs. Bacteria isolated were characterized according to microbiological standards. Factors such as temperature and distance were considered as variable in this study. The isolation was carried out at different temperatures such as 27oC, 31oC and 29oC and at various distances of 100meters and 200meters away from the slaughter sites. Result obtained showed that strains of Staphylococcus aureus, Escherichia coli, Bacillus subtilis, Lactobacillus alimentarius and Micrococcus sp. were identified. The total viable counts showed that more microorganisms were present in the morning while the least viable count of 388cfu was recorded in the evening period of this study. This study also showed that more microbial loads were recorded the further the distance is to the slaughter site. Conclusively, the array of bacteria isolated suggests that abattoir sites may be a potential source of pathogenic organisms to commuters if located within residential environment.

Keywords: Abattoir, Aerosol, Bacterial Composition, Environment.

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174 Neutralization of Alkaline Waste-Waters using a Blend of Microorganisms

Authors: Rita Kumar, Alka Sharma, Purnima Dhall, Niha M. Kulshreshtha, Anil Kumar

Abstract:

The efficient operation of any biological treatment process requires pre-treatment of incompatible pollutants such as acids, bases, oil, toxic substances, etc. which hamper the treatment of other major components which are otherwise degradable. The pre-treatment of alkaline waste-waters, generated from various industries like textile, paper & pulp, potato-processing industries, etc., having a pH of 10 or higher, is essential. The pre-treatment, i.e., neutralization of such alkaline waste-waters can be achieved by chemical as well as biological means. However, the biological pretreatment offers better package over the chemical means by being safe and economical. The biological pre-treatment can be accomplished by using a blend of microorganisms able to withstand such harsh alkaline conditions. In the present study, for the proper pre-treatment of alkaline waste-waters, a package of alkalophilic bacteria is formulated to neutralise the alkaline pH of the industrial waste-waters. The developed microbial package is cost-effective as well as environmental friendly.

Keywords: alkaline, alkalophilic bacteria, biological, pollutants, textile.

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173 Microencapsulation of Probiotic, Evaluation for Viability and Cytotoxic Activities of Its Postbiotic Metabolites on MCF-7 Breast Cancer Cell Line

Authors: N. V. Enwuru, B. Nkeki, E. A. Adekoya, O. A. Adebesin, B. O. Ojo, R. F. Peters, V. A. Aikhomu, U. E. Mendie, O. Akinloye

Abstract:

Awareness about probiotic health benefits is increasing tremendously. However, cell viability is often low due to harsh conditions exposed during processing, handling, storage, and gastrointestinal transit. Thus, encapsulation is a promising technique that increases cell viability. The study aims to encapsulate the probiotic, evaluate its viability and cytotoxic activity of its postbiotic on the Michigan Cancer Foundation (MCF)-7 breast cancer cell line. Human and animal raw milk was sampled for lactic acid bacteria. Isolated bacteria were identified using conventional and VITEK 2 systems. The identified bacteria were encapsulated using the spray-drying method. The free and encapsulated probiotic cells were exposed to simulated gastric intestinal (SGI) fluid conditions and different storage conditions for their viability. The properties of the formed probiotic granules, their disintegration time, and the weight uniformity of the microcapsules were tested. Furthermore, the postbiotic of the free cells was extracted, and its cytotoxic effect on the MCF-7 breast cancer cell line was tested through [3-(4,5-dimethylthiazolyl-2)-2,5 diphenyltetrazolium bromide] (MTT) assay. The bacteria isolated were identified as Lactobacillus plantarum. The size of the formed probiotic granules ranges within 0.71-1.00 mm in diameter, and disintegration time ranges from 2.14 ± 0.045 to 2.91 ± 0.293 minutes, while the average weight is 502.1 mg. The viability of encapsulated cells stored at refrigerated condition (4oC) was higher than that of cells stored at room temperature (25 oC). The encapsulated probiotic cells exhibited better viability after exposure to SGI solution at different pH levels than free cells. The Postbiotic Metabolites (PM) of L. plantarum produced a cytotoxic effect that shows significant activity similar to 5FU, a standard antineoplastic agent. The inhibition concentration of 50% growth (IC50) of postbiotic metabolite was consistent with the IC50 of the positive control (Cisplatin). Lactobacillus plantarum postbiotic exhibited a cytotoxic effect on the MCF-7 breast cancer cell line and could be used as combined adjuvant therapy in breast cancer management. The microencapsulation technique protects the probiotics and maintains their viability.

Keywords: Cytotoxicity effect, encapsulation, postbiotic, probiotic.

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172 Phytoadaptation in Desert Soil Prediction Using Fuzzy Logic Modeling

Authors: S. Bouharati, F. Allag, M. Belmahdi, M. Bounechada

Abstract:

In terms of ecology forecast effects of desertification, the purpose of this study is to develop a predictive model of growth and adaptation of species in arid environment and bioclimatic conditions. The impact of climate change and the desertification phenomena is the result of combined effects in magnitude and frequency of these phenomena. Like the data involved in the phytopathogenic process and bacteria growth in arid soil occur in an uncertain environment because of their complexity, it becomes necessary to have a suitable methodology for the analysis of these variables. The basic principles of fuzzy logic those are perfectly suited to this process. As input variables, we consider the physical parameters, soil type, bacteria nature, and plant species concerned. The result output variable is the adaptability of the species expressed by the growth rate or extinction. As a conclusion, we prevent the possible strategies for adaptation, with or without shifting areas of plantation and nature adequate vegetation.

Keywords: Climate changes, dry soil, Phytopathogenicity, Predictive model, Fuzzy logic.

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171 Re-Examination of Louis Pasteur’s S-Shaped Flask Experiment

Authors: Ming-Hua Fu

Abstract:

No negative control nor control to prevent microbes from escaping was set when the S-shaped flask experiments were performed by Pasteur. Microscope was not used to observe the media in the flasks. Louis Pasteur’s S-shaped flask experiment was re-examined by using U-shaped flasks, modified S-shaped flasks and microscope. A mixture of microbes was isolated from the room air, from which one rod-shaped Bacillus species with proposed name Bacillus gaso-mobilis sp nov and one grape-shaped Staphylococcus species with proposed name of Staphylococcus gaso-mobilis sp nov were identified. Their penicillin and ampicillin resistant strains containing plasmids were isolated. These bacteria could change color, produce odor and automatically move in the air. They did not form colonies on solid media. They had a high suspension capacity in liquid media. Their light absorbance peaked at the wave length of 320 nm.  It was concluded that there were flaws with Louis Pasteur’s S-shaped flask experiments. 

Keywords: Bacteria, gaso-mobile, re-examine, S-shaped flasks.

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170 The Effect of Nano-Silver Packaging on Quality Maintenance of Fresh Strawberry

Authors: Naser Valipour Motlagh, Majid Aliabadi, Elnaz Rahmani, Samira Ghorbanpour

Abstract:

Strawberry is one of the most favored fruits all along the world. But due to its vulnerability to microbial contamination and short life storage, there are lots of problems in industrial production and transportation of this fruit. Therefore, lots of ideas have tried to increase the storage life of strawberries especially through proper packaging. This paper works on efficient packaging as well. The primary material used is produced through simple mixing of low-density polyethylene (LDPE) and silver nanoparticles in different weight fractions of 0.5 and 1% in presence of dicumyl peroxide as a cross-linking agent. Final packages were made in a twin-screw extruder. Then, their effect on the quality maintenance of strawberry is evaluated. The SEM images of nano-silver packages show the distribution of silver nanoparticles in the packages. Total bacteria count, mold, yeast and E. coli are measured for microbial evaluation of all samples. Texture, color, appearance, odor, taste and total acceptance of various samples are evaluated by trained panelists and based on 9-point hedonic scale method. The results show a decrease in total bacteria count and mold in nano-silver packages compared to the samples packed in polyethylene packages for the same storage time. The optimum concentration of silver nanoparticles for the lowest bacteria count and mold is predicted to be around 0.5% which has attained the most acceptance from the panelist as well. Moreover, organoleptic properties of strawberry are preserved for a longer period in nano-silver packages. It can be concluded that using nano-silver particles in strawberry packages has improved the storage life and quality maintenance of the fruit.

Keywords: Antimicrobial properties, polyethylene, silver nanoparticles, strawberry.

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169 Optimization of Growth of Rhodobacter Sphaeroides Using Mixed Volatile Fatty Acidsby Response Surface Methodology

Authors: R.Sangeetha, T.Karunanithi

Abstract:

A combination of photosynthetic bacteria along with anaerobic acidogenic bacteria is an ideal option for efficient hydrogen production. In the present study, the optimum concentration of substrates for the growth of Rhodobacter sphaeroides was found by response surface methodology. The optimum combination of three individual fatty acids was determined by Box Behnken design. Increase of volatile fatty acid concentration decreased the growth. Combination of sodium acetate and sodium propionate was most significant for the growth of the organism. The results showed that a maximum biomass concentration of 0.916 g/l was obtained when the concentrations of acetate, propionate and butyrate were 0.73g/l,0.99g/l and 0.799g/l, respectively. The growth was studied under an optimum concentration of volatile fatty acids and at a light intensity of 3000 lux, initial pH of 7 and a temperature of 35°C.The maximum biomass concentration of 0.92g/l was obtained which verified the practicability of this optimization.

Keywords: Biohydrogen, Response Surface Methodology, Rhodobacter sphaeroides, Volatile fatty acid

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168 Antimicrobial Agents Produced by Yeasts

Authors: T. Buyuksirit, H. Kuleasan

Abstract:

Natural antimicrobials are used to preserve foods that can be found in plants, animals, and microorganisms. Antimicrobial substances are natural or artificial agents that produced by microorganisms or obtained semi/total chemical synthesis are used at low concentrations to inhibit the growth of other microorganisms. Food borne pathogens and spoilage microorganisms are inactivated by the use of antagonistic microorganisms and their metabolites. Yeasts can produce toxic proteins or glycoproteins (toxins) that cause inhibition of sensitive bacteria and yeast species. Antimicrobial substance producing phenotypes belonging different yeast genus were isolated from different sources. Toxins secreted by many yeast strains inhibiting the growth of other yeast strains. These strains show antimicrobial activity, inhibiting the growth of mold and bacteria. The effect of antimicrobial agents produced by yeasts can be extremely fast, and therefore may be used in various treatment procedures. Rapid inhibition of microorganisms is possibly caused by microbial cell membrane lipopolysaccharide binding and in activation (neutralization) effect. Antimicrobial agents inhibit the target cells via different mechanisms of action.

Keywords: Antimicrobial agents, Glycoprotein, Toxic protein, Yeast.

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167 In vitro Study of Antibacterial Activity of Cymbopogon citratus

Authors: C.K. Hindumathy

Abstract:

Alcohol and water extracts of Cymbopogon citratus was investigated for anti-bacterial properties and phytochemical constituents. The extract was screened against four gram-negative bacteria Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus vulgaris) and two grampositive bacteria Bacillus subtilis and Staphylococcus aureus at four different concentrations (1:1, 1:5, 1:10 and 1:20) using disc diffusion method. The antibacterial examination was by disc diffusion techniques, while the photochemical constituents were investigated using standard chemical methods. Results showed that the extracts inhibited the growth of standard and local strains of the organisms used. The treatments were significantly different (P = 0.05). The minimum inhibitory concentration of the extracts against the tested microorganisms ranged between 150mg/ml and 50mg/ml. The alcohol extracts were found to be generally more effective than the water extract. The photochemical analysis revealed the presence of alkaloids and phenol but absence of cardiac and cyanogenic glycosides. The presence of alkaloid and phenols were inferred as being responsible for the anti-bacterial properties of the extracts.

Keywords: Cymbopogon citratus; gram negative and grampositive

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166 Isolation and Screening of Laccase Producing Basidiomycetes via Submerged Fermentations

Authors: Mun Yee Chan, Sin Ming Goh, Lisa Gaik Ai Ong

Abstract:

Approximately 10,000 different types of dyes and pigments are being used in various industrial applications yearly, which include the textile and printing industries. However, these dyes are difficult to degrade naturally once they enter the aquatic system. Their high persistency in natural environment poses a potential health hazard to all form of life. Hence, there is a need for alternative dye removal strategy in the environment via bioremediation. In this study, fungi laccase is investigated via commercial agar dyes plates and submerged fermentation to explore the application of fungi laccase in textile dye wastewater treatment. Two locally isolated basidiomycetes were screened for laccase activity using media added with commercial dyes such as 2, 2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS), guaiacol and Remazol Brillant Blue R (RBBR). Isolate TBB3 (1.70±0.06) and EL2 (1.78±0.08) gave the highest results for ABTS plates with the appearance of greenish halo on around the isolates. Submerged fermentation performed on Isolate TBB3 with the productivity 3.9067 U/ml/day, whereas the laccase activity for Isolate EL2 was much lower (0.2097 U/ml/day). As isolate TBB3 showed higher laccase production, it was subjected to molecular characterization by DNA isolation, PCR amplification and sequencing of ITS region of nuclear ribosomal DNA. After being compared with other sequences in National Center for Biotechnology Information (NCBI database), isolate TBB3 is probably from species Trametes hirsutei. Further research work can be performed on this isolate by upscale the production of laccase in order to meet the demands of the requirement for higher enzyme titer for the bioremediation of textile dyes.

Keywords: Bioremediation, dyes, fermentation, laccase.

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165 Emergence of Fluoroquinolone Resistance in Pigs, Nigeria

Authors: Igbakura I. Luga, Alex A. Adikwu

Abstract:

A comparison of resistance to quinolones was carried out on isolates of Shiga toxin-producing Escherichia coliO157:H7 from cattle and mecA and nuc genes harbouring Staphylococcus aureus from pigs. The isolates were separately tested in the first and current decades of the 21st century. The objective was to demonstrate the dissemination of resistance to this frontline class of antibiotic by bacteria from food animals and bring to the limelight the spread of antibiotic resistance in Nigeria. A total of 10 isolates of the E. coli O157:H7 and 9 of mecA and nuc genes harbouring S. aureus were obtained following isolation, biochemical testing, and serological identification using the Remel Wellcolex E. coli O157:H7 test. Shiga toxin-production screening in the E. coli O157:H7 using the verotoxin E. coli reverse passive latex agglutination (VTEC-RPLA) test; and molecular identification of the mecA and nuc genes in S. aureus. Detection of the mecA and nuc genes were carried out using the protocol by the Danish Technical University (DTU) using the following primers mecA-1:5'-GGGATCATAGCGTCATTATTC-3', mecA-2: 5'-AACGATTGTGACACGATAGCC-3', nuc-1: 5'-TCAGCAAATGCATCACAAACAG-3', nuc-2: 5'-CGTAAATGCACTTGCTTCAGG-3' for the mecA and nuc genes, respectively. The nuc genes confirm the S. aureus isolates and the mecA genes as being methicillin-resistant and so pathogenic to man. The fluoroquinolones used in the antibiotic resistance testing were norfloxacin (10 µg) and ciprofloxacin (5 µg) in the E. coli O157:H7 isolates and ciprofloxacin (5 µg) in the S. aureus isolates. Susceptibility was tested using the disk diffusion method on Muller-Hinton agar. Fluoroquinolone resistance was not detected from isolates of E. coli O157:H7 from cattle. However, 44% (4/9) of the S. aureus were resistant to ciprofloxacin. Resistance of up to 44% in isolates of mecA and nuc genes harbouring S. aureus is a compelling evidence for the rapid spread of antibiotic resistance from bacteria in food animals from Nigeria. Ciprofloxacin is the drug of choice for the treatment of Typhoid fever, therefore widespread resistance to it in pathogenic bacteria is of great public health significance. The study concludes that antibiotic resistance in bacteria from food animals is on the increase in Nigeria. The National Food and Drug Administration and Control (NAFDAC) agency in Nigeria should implement the World Health Organization (WHO) global action plan on antimicrobial resistance. A good starting point can be coordinating the WHO, Office of International Epizootics (OIE), Food and Agricultural Organization (FAO) tripartite draft antimicrobial resistance monitoring and evaluation (M&E) framework in Nigeria.

Keywords: Fluoroquinolone, Nigeria, resistance, Staphylococcus aureus.

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164 From Primer Generation to Chromosome Identification: A Primer Generation Genotyping Method for Bacterial Identification and Typing

Authors: Wisam H. Benamer, Ehab A. Elfallah, Mohamed A. Elshaari, Farag A. Elshaari

Abstract:

A challenge for laboratories is to provide bacterial identification and antibiotic sensitivity results within a short time. Hence, advancement in the required technology is desirable to improve timing, accuracy and quality. Even with the current advances in methods used for both phenotypic and genotypic identification of bacteria the need is there to develop method(s) that enhance the outcome of bacteriology laboratories in accuracy and time. The hypothesis introduced here is based on the assumption that the chromosome of any bacteria contains unique sequences that can be used for its identification and typing. The outcome of a pilot study designed to test this hypothesis is reported in this manuscript. Methods: The complete chromosome sequences of several bacterial species were downloaded to use as search targets for unique sequences. Visual basic and SQL server (2014) were used to generate a complete set of 18-base long primers, a process started with reverse translation of randomly chosen 6 amino acids to limit the number of the generated primers. In addition, the software used to scan the downloaded chromosomes using the generated primers for similarities was designed, and the resulting hits were classified according to the number of similar chromosomal sequences, i.e., unique or otherwise. Results: All primers that had identical/similar sequences in the selected genome sequence(s) were classified according to the number of hits in the chromosomes search. Those that were identical to a single site on a single bacterial chromosome were referred to as unique. On the other hand, most generated primers sequences were identical to multiple sites on a single or multiple chromosomes. Following scanning, the generated primers were classified based on ability to differentiate between medically important bacterial and the initial results looks promising. Conclusion: A simple strategy that started by generating primers was introduced; the primers were used to screen bacterial genomes for match. Primer(s) that were uniquely identical to specific DNA sequence on a specific bacterial chromosome were selected. The identified unique sequence can be used in different molecular diagnostic techniques, possibly to identify bacteria. In addition, a single primer that can identify multiple sites in a single chromosome can be exploited for region or genome identification. Although genomes sequences draft of isolates of organism DNA enable high throughput primer design using alignment strategy, and this enhances diagnostic performance in comparison to traditional molecular assays. In this method the generated primers can be used to identify an organism before the draft sequence is completed. In addition, the generated primers can be used to build a bank for easy access of the primers that can be used to identify bacteria.

Keywords: Bacteria chromosome, bacterial identification, sequence, primer generation.

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163 The Use of Acid-Aluminium Tolerant Bradyrhizobium japonicum Formula for

Authors: Nisa Rachmania Mubarik, Tedja Imas, Aris Tri Wahyudi , Triadiati , Suharyanto, Happy Widiastuti

Abstract:

Land with low pH soil spread widely in Indonesia can be used for soybean (Glycine max) cultivation, however the production is low. The use of acid tolerant soybean and acidaluminium tolerant nitrogen-fixing bacteria formula was an alternative way to increase soybean productivity on acid soils. Bradyrhizobium japonicum is one of the nitrogen fixing bacteria which can symbiose with soybean plants through root nodule formation. Most of the nitrogen source required by soybean plants can be provided by this symbiosis. This research was conducted to study the influence of acid-aluminium tolerant B. japonicum strain BJ 11 formula using peat as carrier on growth of Tanggamus and Anjasmoro cultivar soybean planted on acid soil fields (pH 5.0- 5.5). The results showed that the inoculant was able to increase the growth and production of soybean which were grown on fields acid soil at Sukadana (Lampung) and Tanah Laut (South Kalimantan), Indonesia.

Keywords: Bradyrhizobium japonicum, acid-aluminium tolerant mutant, Tanggamus cultivar soybean, acid soils

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162 Identification and Characterization of Heavy Metal Resistant Bacteria from the Klip River

Authors: P. Chihomvu, P. Stegmann, M. Pillay

Abstract:

Pollution of the Klip River has caused microorganisms inhabiting it to develop protective survival mechanisms. This study isolated and characterized the heavy metal resistant bacteria in the Klip River. Water and sediment samples were collected from six sites along the course of the river. The pH, turbidity, salinity, temperature and dissolved oxygen were measured in-situ. The concentrations of six heavy metals (Cd, Cu, Fe, Ni, Pb and Zn) of the water samples were determined by atomic absorption spectroscopy. Biochemical and antibiotic profiles of the isolates were assessed using the API 20E® and Kirby Bauer Method. Growth studies were carried out using spectrophotometric methods. The isolates were identified using 16SrDNA sequencing. The uppermost part of the Klip River with the lowest pH had the highest levels of heavy metals. Turbidity, salinity and specific conductivity increased measurably at Site 4 (Henley on Klip Weir). MIC tests showed that 16 isolates exhibited high iron and lead resistance. Antibiotic susceptibility tests revealed that the isolates exhibited multitolerances to drugs such as Tetracycline, Ampicillin, and Amoxicillin.

Keywords: Klip River, heavy metals, resistance.

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161 The Antibacterial and Anticancer Activity of Marine Actinomycete Strain HP411 Isolated in the Northern Coast of Vietnam

Authors: Huyen T. Pham, Nhue P. Nguyen, Tien Q. Phi, Phuong T. Dang, Hy G. Le

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Since the marine environmental conditions are extremely different from the other ones, marine actinomycetes might produce novel bioactive compounds. Therefore, actinomycete strains were screened from marine water and sediment samples collected from the coastal areas of Northern Vietnam. Ninety-nine actinomycete strains were obtained on starch-casein agar media by dilution technique, only seven strains, named HP112, HP12, HP411, HPN11, HP 11, HPT13 and HPX12, showed significant antibacterial activity against both gram-positive and gram-negative bacteria (Bacillus subtilis ATCC 6633, Staphylococcus epidemidis ATCC 12228, Escherichia coli ATCC 11105). Further studies were carried out with the most active HP411 strain against Candida albicans ATCC 10231. This strain could grow rapidly on starch casein agar and other media with high salt containing 7-10% NaCl at 28-30oC. Spore-chain of HP411 showed an elongated and circular shape with 10 to 30 spores/chain. Identification of the strain was carried out by employing the taxonomical studies including the 16S rRNA sequence. Based on phylogenetic and phenotypic evidence it is proposed that HP411 to be belongs to species Streptomyces variabilis. The potent of the crude extract of fermentation broth of HP411 that are effective against wide range of pathogens: both grampositive, gram-negative and fungi. Further studies revealed that the crude extract HP411 could obtain the anticancer activity for cancer cell lines: Hep-G2 (liver cancer cell line); RD (cardiac and skeletal muscle letters cell line); FL (membrane of the uterus cancer cell line). However, the actinomycetes from marine ecosystem will be useful for the discovery of new drugs in the future.

Keywords: Marine actinomycetes, antibacterial, anticancer, Streptomyces variabilis.

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