Search results for: coliform bacteria
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1518

Search results for: coliform bacteria

708 Isolation and Characterization of Salt-Tolerance of Rhizobia under the Effects of Salinity

Authors: Sarra Sobti, Baelhadj Hamdi-Aïssa

Abstract:

The bacteria of the soil, usually called rhizobium, have a considerable importance in agriculture because of their capacity to fix the atmospheric nitrogen in symbiosis with the plants of the family of legumes. The present work was to study the effect of the salinity on growth and nodulation of alfalfa-rhizobia symbiosis at different agricultural experimental sites in Ouargla. The experiment was conducted in 3 steps. The first one was the isolation and characterization of the Rhizobia; next, the evolution of the isolates tolerance to salinity at three levels of NaCl (6, 8,12 and 16 g/L); and the last step was the evolution of the tolerance on symbiotic characteristics. The results showed that the phenotypic characterizations behave practically as Rhizobia spp, and the effects of salinity affect the symbiotic process. The tolerance to high levels of salinity and the survival and persistence in severe and harsh desert conditions make these rhizobia highly valuable inoculums to improve productivity of the leguminous plants cultivated under extreme environments.

Keywords: rhizobia, symbiosis, salinity, tolerance, nodulation, soil, Medicago sativa L.

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707 Treatment of Acid Mine Drainage with Modified Fly Ash

Authors: Sukla Saha, Alok Sinha

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Acid mine drainage (AMD) is the generation of acidic water from active as well as abandoned mines. AMD generates due to the oxidation of pyrites present in the rock in mining areas. Sulfur oxidizing bacteria such as Thiobacillus ferrooxidans acts as a catalyst in this oxidation process. The characteristics of AMD is extreme low pH (2-3) with elevated concentration of different heavy metals such as Fe, Al, Zn, Mn, Cu and Co and anions such sulfate and chloride. AMD contaminate the ground water as well as surface water which leads to the degradation of water quality. Moreover, it carries detrimental effect for aquatic organism and degrade the environment. In the present study, AMD is treated with fly ash, modified with alkaline agent (NaOH). This modified fly ash (MFA) was experimentally proven as a very effective neutralizing agent for the treatment of AMD. It was observed that pH of treated AMD raised to 9.22 from 1.51 with 100g/L of MFA dose. Approximately, 99% removal of Fe, Al, Mn, Cu and Co took place with the same MFA dose. The treated water comply with the effluent discharge standard of (IS: 2490-1981).

Keywords: acid mine drainage, heavy metals, modified fly ash, neutralization

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706 Acute Phase Proteins as Biomarkers of Urinary Tract Infection (UTI) in Dairy Cattle

Authors: Wael El-Deeb

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The present study aimed to investigate the diagnostic importance of acute phase proteins in urinary tract infection (UTI) in cattle. We describe the clinical, bacteriological and biochemical findings in 99 lactating cows. Blood and urine samples from diseased (n=84) and control healthy cows (n=15) were submitted to laboratory investigations. The urine analysis revealed hematuria and pyuria in UTI group. The isolated bacteria were E.coli (43/84) Corynebacterium spp, (31/84), Proteus spp. (6/84) and Streptococcus spp (4/84). The concentrations of Haptoglobin (Hp), serum amyloid A (SAA), α1-Acid glycoprotein (AGP), fibrinogen (Fb), total protein, albumen, and globulin were higher in cows with UTI when compared to healthy ones. Fifty-one of 84 cows with UTI were successfully treated. The levels of Hp, SAA, AGP, total protein, and globulin were associated with the odds of treatment failure. Conclusively, acute phase proteins could be used as diagnostic and prognostic biomarkers in cows with UTI.

Keywords: cows, urinary, infections, haptoglobin, serum Amyloid A

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705 Effect of Zinc Oxide Nanoparticles along with Sodium Hydroxide on Self-Cleaning and Antibacterial Properties of Polyethylene Terephthalate

Authors: Mohammad Mirjalili, Maryam Mohammdi, Loghman Karimi

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In this study, synthesis of zinc oxide nanoparticles was carried out along with the hydrolysis of Polyethylene terephthalate using sodium hydroxide to increase the surface activity and enhance the nanoparticles adsorption. The polyester fabrics were treated with zinc acetate and sodium hydroxide at ultrasound bath, resulting in the formation of ZnO nanospheres. The presence of zinc oxide on the surface of the polyethylene terephthalate was confirmed by scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS). The self-cleaning property of treated polyethylene terephthalate was evaluated through discoloring methylene blue stain under sunlight irradiation. The antibacterial activities of the samples against two common pathogenic bacteria including Escherichia coli and Staphylococcus aureus were also assessed. The results indicated that the photocatalytic and antibacterial activities of the ultrasound treated polyethylene terephthalate improved significantly.

Keywords: zinc oxide, polyethylene terephthalate, self-cleaning, antibacterial

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704 Microbial Fuel Cells: Performance and Applications

Authors: Andrea Pietrelli, Vincenzo Ferrara, Bruno Allard, Francois Buret, Irene Bavasso, Nicola Lovecchio, Francesca Costantini, Firas Khaled

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This paper aims to show some applications of microbial fuel cells (MFCs), an energy harvesting technique, as clean power source to supply low power device for application like wireless sensor network (WSN) for environmental monitoring. Furthermore, MFC can be used directly as biosensor to analyse parameters like pH and temperature or arranged in form of cluster devices in order to use as small power plant. An MFC is a bioreactor that converts energy stored in chemical bonds of organic matter into electrical energy, through a series of reactions catalysed by microorganisms. We have developed a lab-scale terrestrial microbial fuel cell (TMFC), based on soil that acts as source of bacteria and flow of nutrient and a lab-scale waste water microbial fuel cell (WWMFC), where waste water acts as flow of nutrient and bacteria. We performed large series of tests to exploit the capability as biosensor. The pH value has strong influence on the open circuit voltage (OCV) delivered from TMFCs. We analyzed three condition: test A and B were filled with same soil but changing pH from 6 to 6.63, test C was prepared using a different soil with a pH value of 6.3. Experimental results clearly show how with higher pH value a higher OCV was produced; indeed reactors are influenced by different values of pH which increases the voltage in case of a higher pH value until the best pH value of 7 is achieved. The influence of pH on OCV of lab-scales WWMFC was analyzed at pH value of 6.5, 7, 7.2, 7.5 and 8. WWMFCs are influenced from temperature more than TMFCs. We tested the power performance of WWMFCs considering four imposed values of ambient temperature. Results show how power performance increase proportionally with higher temperature values, doubling the output power from 20° to 40°. The best value of power produced from our lab-scale TMFC was equal to 310 μW using peaty soil, at 1KΩ, corresponding to a current of 0.5 mA. A TMFC can supply proper energy to low power devices of a WSN by means of the design of three stages scheme of an energy management system, which adapts voltage level of TMFC to those required by a WSN node, as 3.3V. Using a commercial DC/DC boost converter, that needs an input voltage of 700 mV, the current source of 0.5 mA, charges a capacitor of 6.8 mF until it will have accumulated an amount of charge equal to 700 mV in a time of 10 s. The output stage includes an output switch that close the circuit after a time of 10s + 1.5ms because the converter can boost the voltage from 0.7V to 3.3V in 1.5 ms. Furthermore, we tested in form of clusters connected in series up to 20 WWMFCs, we have obtained a high voltage value as output, around 10V, but low current value. MFC can be considered a suitable clean energy source to be used to supply low power devices as a WSN node or to be used directly as biosensor.

Keywords: energy harvesting, low power electronics, microbial fuel cell, terrestrial microbial fuel cell, waste-water microbial fuel cell, wireless sensor network

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703 Relative Expression and Detection of MUB Adhesion Domains and Plantaricin-Like Bacteriocin among Probiotic Lactobacillus plantarum-Group Strains Isolated from Fermented Foods

Authors: Sundru Manjulata Devi, Prakash M. Halami

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The immemorial use of fermented foods from vegetables, dairy and other biological sources are of great demand in India because of their health benefits. However, the diversity of Lactobacillus plantarum group (LPG) of vegetable origin has not been revealed yet, particularly with reference to their probiotic functionalities. In the present study, the different species of probiotic Lactobacillus plantarum group (LPG) i.e., L. plantarum subsp. plantarum MTCC 5422 (from fermented cereals), L. plantarum subsp. argentoratensis FG16 (from fermented bamboo shoot) and L. paraplantarum MTCC 9483 (from fermented gundruk) (as characterized by multiplex recA PCR assay) were considered to investigate their relative expression of MUB domains of mub gene (mucin binding protein) by Real time PCR. Initially, the allelic variation in the mub gene was assessed and found to encode three different variants (Type I, II and III). All the three types had 8, 9 and 10 MUB domains respectively (as analysed by Pfam database) and were found to be responsible for adhesion of bacteria to the host intestinal epithelial cells. These domains either get inserted or deleted during speciation or evolutionary events and lead to divergence. The reverse transcriptase qPCR analysis with mubLPF1+R1 primer pair supported variation in amplicon sizes with 300, 500 and 700 bp among different LPG strains. The relative expression of these MUB domains significantly unregulated in the presence of 1% mucin in overnight grown cultures. Simultaneously, the mub gene expressed efficiently by 7 fold in the culture L. paraplantarum MTCC 9483 with 10 MUB domains. An increase in the expression levels for L. plantarum subsp. plantarum MTCC 5422 and L. plantarum subsp. argentoratensis FG16 (MCC 2974) with 9 and 8 repetitive domains was around 4 and 2 fold, respectively. The detection and expression of an integrase (int) gene in the upstream region of mub gene reveals the excision and integration of these repetitive domains. Concurrently, an in vitro adhesion assay to mucin and exclusion of pathogens (such as Listeria monocytogenes and Micrococcus leuteus) was investigated and observed that the L. paraplantarum MTCC 9483 with more adhesion domains has more ability to adhere to mucin and inhibited the growth of pathogens. The production and expression of plantaricin-like bacteriocin (plnNC8 type) in MTCC 9483 suggests the pathogen inhibition. Hence, the expression of MUB domains can act as potential biomarkers in the screening of a novel probiotic LPG strain with adherence property. The present study provides a platform for an easy, rapid, less time consuming, low-cost methodology for the detection of potential probiotic bacteria. It was known that the traditional practices followed in the preparation of fermented bamboo shoots/gundruk/cereals of Indian foods contain different kinds of neutraceuticals for functional food and novel compounds with health promoting factors. In future, a detailed study of these food products can add more nutritive value, consumption and suitable for commercialization.

Keywords: adhesion gene, fermented foods, MUB domains, probiotics

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702 One Step Green Synthesis of Silver Nanoparticles and Their Biological Activity

Authors: Samy M. Shaban, Ismail Aiad, Mohamed M. El-Sukkary, E. A. Soliman, Moshira Y. El-Awady

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In situ and green synthesis of cubic and spherical silver nanoparticles were developed using sun light as reducing agent in the presence of newly prepared cationic surfactant which acting as capping agents. The morphology of prepared silver nanoparticle was estimated by transmission electron microscope (TEM) and the size distribution determined by dynamic light scattering (DLS). The hydrophobic chain length of the prepared surfactant effect on the stability of the prepared silver nanoparticles as clear from zeta-potential values. Also by increasing chain length of the used capping agent the amount of formed nanoparticle increase as indicated by increasing the absorbance. Both prepared surfactants and surfactants capping silver nanoparticles showed high antimicrobial activity against gram positive and gram-negative bacteria.

Keywords: photosynthesis, hexaonal shapes, zetapotential, biological activity

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701 Diversity of Microbial Ground Improvements

Authors: V. Ivanov, J. Chu, V. Stabnikov

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Low cost, sustainable, and environmentally friendly microbial cements, grouts, polysaccharides and bioplastics are useful in construction and geotechnical engineering. Construction-related biotechnologies are based on activity of different microorganisms: urease-producing, acidogenic, halophilic, alkaliphilic, denitrifying, iron- and sulphate-reducing bacteria, cyanobacteria, algae, microscopic fungi. The bio-related materials and processes can be used for the bioaggregation, soil biogrouting and bioclogging, biocementation, biodesaturation of water-satured soil, bioencapsulation of soft clay, biocoating, and biorepair of the concrete surface. Altogether with the most popular calcium- and urea based biocementation, there are possible and often are more effective such methods of ground improvement as calcium- and magnesium based biocementation, calcium phosphate strengthening of soil, calcium bicarbonate biocementation, and iron- or polysaccharide based bioclogging. The construction-related microbial biotechnologies have a lot of advantages over conventional construction materials and processes.

Keywords: ground improvement, biocementation, biogrouting, microorganisms

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700 Artificial Cells Capable of Communication by Using Polymer Hydrogel

Authors: Qi Liu, Jiqin Yao, Xiaohu Zhou, Bo Zheng

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The first artificial cell was produced by Thomas Chang in the 1950s when he was trying to make a mimic of red blood cells. Since then, many different types of artificial cells have been constructed from one of the two approaches: a so-called bottom-up approach, which aims to create a cell from scratch, and a top-down approach, in which genes are sequentially knocked out from organisms until only the minimal genome required for sustaining life remains. In this project, bottom-up approach was used to build a new cell-free expression system which mimics artificial cell that capable of protein expression and communicate with each other. The artificial cells constructed from the bottom-up approach are usually lipid vesicles, polymersomes, hydrogels or aqueous droplets containing the nucleic acids and transcription-translation machinery. However, lipid vesicles based artificial cells capable of communication present several issues in the cell communication research: (1) The lipid vesicles normally lose the important functions such as protein expression within a few hours. (2) The lipid membrane allows the permeation of only small molecules and limits the types of molecules that can be sensed and released to the surrounding environment for chemical communication; (3) The lipid vesicles are prone to rupture due to the imbalance of the osmotic pressure. To address these issues, the hydrogel-based artificial cells were constructed in this work. To construct the artificial cell, polyacrylamide hydrogel was functionalized with Acrylate PEG Succinimidyl Carboxymethyl Ester (ACLT-PEG2000-SCM) moiety on the polymer backbone. The proteinaceous factors can then be immobilized on the polymer backbone by the reaction between primary amines of proteins and N-hydroxysuccinimide esters (NHS esters) of ACLT-PEG2000-SCM, the plasmid template and ribosome were encapsulated inside the hydrogel particles. Because the artificial cell could continuously express protein with the supply of nutrients and energy, the artificial cell-artificial cell communication and artificial cell-natural cell communication could be achieved by combining the artificial cell vector with designed plasmids. The plasmids were designed referring to the quorum sensing (QS) system of bacteria, which largely relied on cognate acyl-homoserine lactone (AHL) / transcription pairs. In one communication pair, “sender” is the artificial cell or natural cell that can produce AHL signal molecule by synthesizing the corresponding signal synthase that catalyzed the conversion of S-adenosyl-L-methionine (SAM) into AHL, while the “receiver” is the artificial cell or natural cell that can sense the quorum sensing signaling molecule form “sender” and in turn express the gene of interest. In the experiment, GFP was first immobilized inside the hydrogel particle to prove that the functionalized hydrogel particles could be used for protein binding. After that, the successful communication between artificial cell-artificial cell and artificial cell-natural cell was demonstrated, the successful signal between artificial cell-artificial cell or artificial cell-natural cell could be observed by recording the fluorescence signal increase. The hydrogel-based artificial cell designed in this work can help to study the complex communication system in bacteria, it can also be further developed for therapeutic applications.

Keywords: artificial cell, cell-free system, gene circuit, synthetic biology

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699 The Impact of Different Rhizobium leguminosarum Strains on the Protein Content of Peas and Broad Beans

Authors: Alise Senberga, Laila Dubova, Liene Strauta, Ina Alsina, Ieva Erdberga

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Legume symbiotic relationship with nitrogen fixating bacteria Rhizobim leguminosarum is an important factor used to improve the productivity of legumes, due to the fact that rhizobia can supply plant with the necessary amount of nitrogen. R. leguminosarum strains have shown different activity in fixing nitrogen. Depending on the chosen R. leguminosarum strain, host plant biochemical content can be altered. In this study we focused particularly on the changes in protein content in beans (using two different varieties) and peas (five different varieties) due to the use of several different R. leguminosarum strains (four strains for both beans and peas). Overall, the protein content increase was observed after seed inoculation with R. leguminosarum. Strain and plant cultivar interaction specification was observed. The effect of R. leguminosarum inoculation on the content of protein was dependent on the R. leguminosarum strain used. Plant cultivar also appeared to have a decisive role in protein content formation with the help of R. leguminosaru.

Keywords: legumes, protein content, rhizobia strains, soil

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698 Clustered Regularly Interspaced Short Palindromic Repeats Interference (CRISPRi): An Approach to Inhibit Microbial Biofilm

Authors: Azna Zuberi

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Biofilm is a sessile bacterial accretion in which bacteria adapts different physiological and morphological behavior from planktonic form. It is the root cause of about 80% microbial infections in human. Among them, E. coli biofilms are most prevalent in medical devices associated nosocomial infections. The objective of this study was to inhibit biofilm formation by targeting LuxS gene, involved in quorum sensing using CRISPRi. luxS is a synthase, involved in the synthesis of Autoinducer-2(AI-2), which in turn guides the initial stage of biofilm formation. To implement CRISPRi system, we have synthesized complementary sgRNA to target gene sequence and co-expressed with dCas9. Suppression of luxS was confirmed through qRT-PCR. The effect of luxS gene on biofilm inhibition was studied through crystal violet assay, XTT reduction assay and scanning electron microscopy. We conclude that CRISPRi system could be a potential strategy to inhibit bacterial biofilm through mechanism base approach.

Keywords: biofilm, CRISPRi, luxS, microbial

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697 Virtualization of Biomass Colonization: Potential of Application in Precision Medicine

Authors: Maria Valeria De Bonis, Gianpaolo Ruocco

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Nowadays, computational modeling is paving new design and verification ways in a number of industrial sectors. The technology is ripe to challenge some case in the Bioengineering and Medicine frameworks: for example, looking at the strategical and ethical importance of oncology research, efforts should be made to yield new and powerful resources to tumor knowledge and understanding. With these driving motivations, we approach this gigantic problem by using some standard engineering tools such as the mathematics behind the biomass transfer. We present here some bacterial colonization studies in complex structures. As strong analogies hold with some tumor proliferation, we extend our study to a benchmark case of solid tumor. By means of a commercial software, we model biomass and energy evolution in arbitrary media. The approach will be useful to cast virtualization cases of cancer growth in human organs, while augmented reality tools will be used to yield for a realistic aid to informed decision in treatment and surgery.

Keywords: bacteria, simulation, tumor, precision medicine

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696 Cereal Bioproducts Conversion to Higher Value Feed by Using Pediococcus Strains Isolated from Spontaneous Fermented Cereal, and Its Influence on Milk Production of Dairy Cattle

Authors: Vita Krungleviciute, Rasa Zelvyte, Ingrida Monkeviciene, Jone Kantautaite, Rolandas Stankevicius, Modestas Ruzauskas, Elena Bartkiene

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The environmental impact of agricultural bioproducts from the processing of food crops is an increasing concern worldwide. Currently, cereal bran has been used as a low-value ingredient for both human consumption and animal feed. The most popular bioprocessing technologies for cereal bran nutritional and technological functionality increasing are enzymatic processing and fermentation, and the most popular starters in fermented feed production are lactic acid bacteria (LAB) including pediococci. However, the ruminant digestive system is unique, there are billions of microorganisms which help the cow to digest and utilize nutrients in the feed. To achieve efficient feed utilization and high milk yield, the microorganisms must have optimal conditions, and the disbalance of this system is highly undesirable. Pediococcus strains Pediococcus acidilactici BaltBio01 and Pediococcus pentosaceus BaltBio02 from spontaneous fermented rye were isolated (by rep – PCR method), identified, and characterized by their growth (by Thermo Bioscreen C automatic turbidometer), acidification rate (2 hours in 2.5 pH), gas production (Durham method), and carbohydrate metabolism (by API 50 CH test ). Antimicrobial activities of isolated pediococcus against variety of pathogenic and opportunistic bacterial strains previously isolated from diseased cattle, and their resistance to antibiotics were evaluated (EFSA-FEEDAP method). The isolated pediococcus strains were cultivated in barley/wheat bran (90 / 10, m / m) substrate, and developed supplements, with high content of valuable pediococcus, were used for Lithuanian black and white dairy cows feeding. In addition, the influence of supplements on milk production and composition was determined. Milk composition was evaluated by the LactoScope FTIR” FT1.0. 2001 (Delta Instruments, Holland). P. acidilactici BaltBio01 and P. pentosaceus BaltBio02 demonstrated versatile carbohydrate metabolism, grown at 30°C and 37°C temperatures, and acidic tolerance. Isolated pediococcus strains showed to be non resistant to antibiotics, and having antimicrobial activity against undesirable microorganisms. By barley/wheat bran utilisation using fermentation with selected pediococcus strains, it is possible to produce safer (reduced Enterobacteriaceae, total aerobic bacteria, yeast and mold count) feed stock with high content of pediococcus. Significantly higher milk yield (after 33 days) by using pediococcus supplements mix for dairy cows feeding could be obtained, while similar effect by using separate strains after 66 days of feeding could be achieved. It can be stated that barley/wheat bran could be used for higher value feed production in order to increase milk production. Therefore, further research is needed to identify what is the main mechanism of the positive action.

Keywords: barley/wheat bran, dairy cattle, fermented feed, milk, pediococcus

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695 Characterization of Surface Microstructures on Bio-Based PLA Fabricated with Nano-Imprint Lithography

Authors: D. Bikiaris, M. Nerantzaki, I. Koliakou, A. Francone, N. Kehagias

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In the present study, the formation of structures in poly(lactic acid) (PLA) has been investigated with respect to producing areas of regular, superficial features with dimensions comparable to those of cells or biological macromolecules. Nanoimprint lithography, a method of pattern replication in polymers, has been used for the production of features ranging from tens of micrometers, covering areas up to 1 cm², down to hundreds of nanometers. Both micro- and nano-structures were faithfully replicated. Potentially, PLA has wide uses within biomedical fields, from implantable medical devices, including screws and pins, to membrane applications, such as wound covers, and even as an injectable polymer for, for example, lipoatrophy. The possibility of fabricating structured PLA surfaces, with structures of the dimensions associated with cells or biological macro- molecules, is of interest in fields such as cellular engineering. Imprint-based technologies have demonstrated the ability to selectively imprint polymer films over large areas resulting in 3D imprints over flat, curved or pre-patterned surfaces. Here, we compare nano-patterned with nano-patterned by nanoimprint lithography (NIL) PLA film. A silicon nanostructured stamp (provided by Nanotypos company) having positive and negative protrusions was used to pattern PLA films by means of thermal NIL. The polymer film was heated from 40°C to 60°C above its Tg and embossed with a pressure of 60 bars for 3 min. The stamp and substrate were demolded at room temperature. Scanning electron microscope (SEM) images showed good replication fidelity of the replicated Si stamp. Contact-angle measurements suggested that positive microstructuring of the polymer (where features protrude from the polymer surface) produced a more hydrophilic surface than negative micro-structuring. The ability to structure the surface of the poly(lactic acid), allied to the polymer’s post-processing transparency and proven biocompatibility. Films produced in this were also shown to enhance the aligned attachment behavior and proliferation of Wharton’s Jelly Mesenchymal Stem cells, leading to the observed growth contact guidance. The bacterial attachment patterns of some bacteria, highlighted that the nano-patterned PLA structure can reduce the propensity for the bacteria to attach to the surface, with a greater bactericidal being demonstrated activity against the Staphylococcus aureus cells. These biocompatible, micro- and nanopatterned PLA surfaces could be useful for polymer– cell interaction experiments at dimensions at, or below, that of individual cells. Indeed, post-fabrication modification of the microstructured PLA surface, with materials such as collagen (which can further reduce the hydrophobicity of the surface), will extend the range of applications, possibly through the use of PLA’s inherent biodegradability. Further study is being undertaken to examine whether these structures promote cell growth on the polymer surface.

Keywords: poly(lactic acid), nano-imprint lithography, anti-bacterial properties, PLA

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694 Antibacterial Activity of Green Synthesis Silver Nanoparticles from Moringa Oleifera

Authors: Ali Fadhel Ahmed, Tuqa Abdulkareem Hameed

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Moringa oleifera (leaves and seeds) ethanolic and aqueous extracts were tested for antibacterial activity. The effect of plant extracts on three types of bacterial species: Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae, was investigated. Using the agar well diffusion method, ethanolic extracts of Moringa oleifera demonstrated a significant antibacterial effect on the forty tested bacterial strains. Seed-induced inhibition zones (ethanolic extracts)were ranged from16 to 24 mm in diameter against S. aureus, respectively, whileE. coli and K. pneumonia had no effect. Gram-positive and Gram-negative bacteria were not affected by alcoholic and aqueous plant leaf extracts. The purpose of this present study was to look at the cytotoxic effects of M.Oleifera plant (alcoholic extracts).

Keywords: moringa oleifera, escherichia coli, klebsiella pneumoniae, staphylococcus aureus

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693 Production and Characterization of Silver Doped Hydroxyapatite Thin Films for Biomedical Applications

Authors: C. L Popa, C.S. Ciobanu, S. L. Iconaru, P. Chapon, A. Costescu, P. Le Coustumer, D. Predoi

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In this paper, the preparation and characterization of silver doped hydroxyapatite thin films and their antimicrobial activity characterized is reported. The resultant Ag: HAp films coated on commercially pure Si disks substrates were systematically characterized by Scanning Electron Microscopy (SEM) coupled with X-ray Energy Dispersive Spectroscopy detector (X-EDS), Glow Discharge Optical Emission Spectroscopy (GDOES) and Fourier Transform Infrared spectroscopy (FT-IR). GDOES measurements show that a substantial Ag content has been deposited in the films. The X-EDS and GDOES spectra revealed the presence of a material composed mainly of phosphate, calcium, oxygen, hydrogen and silver. The antimicrobial efficiency of Ag:HAp thin films against Escherichia coli and Staphylococcus aureus bacteria was demonstrated. Ag:HAp thin films could lead to a decrease of infections especially in the case of bone and dental implants by surface modification of implantable medical devices.

Keywords: silver, hydroxyapatite, thin films, GDOES, SEM, FTIR, antimicrobial effect

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692 Probiotics as Therapeutic Agents in the Treatment of Various Diseases: A Literature Review

Authors: K. B. Chathyushya, M. Shiva Prakash, R. Hemalatha

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Introduction: Gastrointestinal (GI) tract has a number of microorganisms (microbiota) that influences the host’s health. The imbalance in the gut microbiota, which is also called as gut dysbiosis, affects human health which causes various metabolic, inflammatory, and infectious diseases. Probiotics play an important role in reinstating the gut balance. Probiotics are involved in the maintenance of healthier gut microbiota and have also been identified as effective adjuvants in insulin resistance therapies. Methods: This paper systematically reviews different randomized, controlled, blinded trials of probiotics for the treatment of various diseases along with the therapeutic or prophylactic properties of probiotic bacteria in different metabolic, inflammatory, infectious and anxiety-related disorders. Conclusion: The present review summarises that probiotics have some considerable effect in the management of various diseases, however, the benefits are strain specific, although more clinical trials are need to be carried out with different probiotic and symbiotic combinations as some probiotics have broad spectrum of benefits and few with specific activity

Keywords: life style diseases, cognition, health, gut dysbiosis, probiotics

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691 Gold Nanoprobes Assay for the Identification of Foodborn Pathogens Such as Staphylococcus aureus, Listeria monocytogenes and Salmonella enteritis

Authors: D. P. Houhoula, J. Papaparaskevas, S. Konteles, A. Dargenta, A. Farka, C. Spyrou, M. Ziaka, S. Koussisis, E. Charvalos

Abstract:

Objectives: Nanotechnology is providing revolutionary opportunities for the rapid and simple diagnosis of many infectious diseases. Staphylococcus aureus, Listeria monocytogenes and Salmonella enteritis are important human pathogens. Diagnostic assays for bacterial culture and identification are time consuming and laborious. There is an urgent need to develop rapid, sensitive, and inexpensive diagnostic tests. In this study, a gold nanoprobe strategy developed and relies on the colorimetric differentiation of specific DNA sequences based approach on differential aggregation profiles in the presence or absence of specific target hybridization. Method: Gold nanoparticles (AuNPs) were purchased from Nanopartz. They were conjugated with thiolated oligonucleotides specific for the femA gene for the identification of members of Staphylococcus aureus, the mecA gene for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus, hly gene encoding the pore-forming cytolysin listeriolysin for the identification of Listeria monocytogenes and the invA sequence for the identification of Salmonella enteritis. DNA isolation from Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis cultures was performed using the commercial kit Nucleospin Tissue (Macherey Nagel). Specifically 20μl of DNA was diluted in 10mMPBS (pH5). After the denaturation of 10min, 20μl of AuNPs was added followed by the annealing step at 58oC. The presence of a complementary target prevents aggregation with the addition of acid and the solution remains pink, whereas in the opposite event it turns to purple. The color could be detected visually and it was confirmed with an absorption spectrum. Results: Specifically, 0.123 μg/μl DNA of St. aureus, L.monocytogenes and Salmonella enteritis was serially diluted from 1:10 to 1:100. Blanks containing PBS buffer instead of DNA were used. The application of the proposed method on isolated bacteria produced positive results with all the species of St. aureus and L. monocytogenes and Salmonella enteritis using the femA, mecA, hly and invA genes respectively. The minimum detection limit of the assay was defined at 0.2 ng/μL of DNA. Below of 0.2 ng/μL of bacterial DNA the solution turned purple after addition of HCl, defining the minimum detection limit of the assay. None of the blank samples was positive. The specificity was 100%. The application of the proposed method produced exactly the same results every time (n = 4) the evaluation was repeated (100% repeatability) using the femA, hly and invA genes. Using the gene mecA for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus the method had a repeatability 50%. Conclusion: The proposed method could be used as a highly specific and sensitive screening tool for the detection and differentiation of Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis. The use AuNPs for the colorimetric detection of DNA targets represents an inexpensive and easy-to-perform alternative to common molecular assays. The technology described here, may develop into a platform that could accommodate detection of many bacterial species.

Keywords: gold nanoparticles, pathogens, nanotechnology, bacteria

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690 Application of Phenol Degrading Microorganisms for the Treatment of Olive Mill Waste (OMW)

Authors: M. A. El-Khateeb

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The growth of the olive oil production in Saudi Arabia peculiarly in Al Jouf region in recent years has been accompanied by an increase in the discharge of associated processing waste. Olive mill waste is produced throughout the extraction of oil from the olive fruit using the traditional mill and press process. Deterioration of the environment due to olive mill disposal wastes is a serious problem. When olive mill waste disposed into the soil, it affects soil quality, soil micro flora, and also toxic to plants. The aim of this work is to isolate microorganism (bacterial or fungal strains) from OMW capable of degrading phenols. Olive mill wastewater, olive mill waste and soil (beside oil production mill) contaminated with olive waste were used for isolation of phenol tolerant microorganisms. Four strains (two fungal and two bacterial) were isolated from olive mill waste. The isolated strains were Candida tropicalis and Phanerochaete chrysosporium (fungal strains) and Bacillus sp. and Rhodococcus sp. (bacterial strains). These strains were able to degrade phenols and could be used for bioremediation of olive mill waste.

Keywords: bioremediation, bacteria, fungi, Sakaka

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689 Microbial Biogeography of Greek Olive Varieties Assessed by Amplicon-Based Metagenomics Analysis

Authors: Lena Payati, Maria Kazou, Effie Tsakalidou

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Table olives are one of the most popular fermented vegetables worldwide, which along with olive oil, have a crucial role in the world economy. They are highly appreciated by the consumers for their characteristic taste and pleasant aromas, while several health and nutritional benefits have been reported as well. Until recently, microbial biogeography, i.e., the study of microbial diversity over time and space, has been mainly associated with wine. However, nowadays, the term 'terroir' has been extended to other crops and food products so as to link the geographical origin and environmental conditions to quality aspects of fermented foods. Taking the above into consideration, the present study focuses on the microbial fingerprinting of the most important olive varieties of Greece with the state-of-the-art amplicon-based metagenomics analysis. Towards this, in 2019, 61 samples from 38 different olive varieties were collected at the final stage of ripening from 13 well spread geographical regions in Greece. For the metagenomics analysis, total DNA was extracted from the olive samples, and the 16S rRNA gene and ITS DNA region were sequenced and analyzed using bioinformatics tools for the identification of bacterial and yeasts/fungal diversity, respectively. Furthermore, principal component analysis (PCA) was also performed for data clustering based on the average microbial composition of all samples from each region of origin. According to the composition, results obtained, when samples were analyzed separately, the majority of both bacteria (such as Pantoea, Enterobacter, Roserbergiella, and Pseudomonas) and yeasts/fungi (such as Aureobasidium, Debaromyces, Candida, and Cladosporium) genera identified were found in all 61 samples. Even though interesting differences were observed at the relative abundance level of the identified genera, the bacterial genus Pantoea and the yeast/fungi genus Aureobasidium were the dominant ones in 35 and 40 samples, respectively. Of note, olive samples collected from the same region had similar fingerprint (genera identified and relative abundance level) regardless of the variety, indicating a potential association between the relative abundance of certain taxa and the geographical region. When samples were grouped by region of origin, distinct bacterial profiles per region were observed, which was also evident from the PCA analysis. This was not the case for the yeast/fungi profiles since 10 out of the 13 regions were grouped together mainly due to the dominance of the genus Aureobasidium. A second cluster was formed for the islands Crete and Rhodes, both of which are located in the Southeast Aegean Sea. These two regions clustered together mainly due to the identification of the genus Toxicocladosporium in relatively high abundances. Finally, the Agrinio region was separated from the others as it showed a completely different microbial fingerprinting. However, due to the limited number of olive samples from some regions, a subsequent PCA analysis with more samples from these regions is expected to yield in a more clear clustering. The present study is part of a bigger project, the first of its kind in Greece, with the ultimate goal to analyze a larger set of olive samples of different varieties and from different regions in Greece in order to have a reliable olives’ microbial biogeography.

Keywords: amplicon-based metagenomics analysis, bacteria, microbial biogeography, olive microbiota, yeasts/fungi

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688 Antimicrobial Activity of Biosynthesized Silver Nanoparticles with Handroanthus Chrysanthus Flower Extract

Authors: Eduardo Padilla, Luis Daniel Rodriguez, Ivan Sanchez, Angelica Sofia Go

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The synthesis and application of metallic nanoparticles have increased in recent years. Biological methods go beyond the chemical and physical synthesis that is expensive and not friendly to the environment. Therefore, in this study, silver nanoparticles were synthesized biologically in an environmentally friendly way by Handroanthus chrysanthus flower aqueous extract (AgNPs) that contains phytochemicals capable of reducing silver nitrate. AgNPs were characterized visually by UV-visible spectroscopy and TEM. The antimicrobial activity of the AgNPs was tested by determining the minimum inhibitory concentration (MIC), and minimal bactericidal concentration (MBC) in Escherichia coli and Staphylococcus aureus strains AgNPs showed potent antimicrobial activity against gram-negative and gram-positive bacteria. MIC and MBC values were as low as 41.6, and 83.2 ug/mL using AgNPs biosynthesized by H. chrysanthus flower extract. This nanoparticle could be the basis for the formulation of disinfectants for use in the food and pharmaceutical industry.

Keywords: antimicrobial, silver nanoparticles, flower extract, Handroanthus

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687 Development of Extemporaneous Pediatric Syrup of Prednisone

Authors: Amel Chenafa, Sihem Boulenouar, Linda Aoued, Imane Sediri, Ismahan Djebbar, Mohamed Adil Selka

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Introduction: The specialties intended for adults are often inadequate marketed for pediatric use, such as for a galenic form or in the dosage. For an industrial, development of a pediatric drug is confronted to various problems. So, the hospital pharmacies have to respond to adaptation needs of pharmaceutical forms for pediatric use. The objective of our work is to develop an oral form of prednisone for pediatric use since no adapted form to children is commercialized. Materials and Methods: Therefore an extemporaneous syrup of prednisone was prepared at the concentration of 0,5mg/ml from 5mg tablets and stored in amber glass bottles. Organoleptic and microbiological stability was studied in two temperatures: 5°C and 25°C, and evaluated at D0, D15, and D30. Results: No organoleptic changes have been detected on the syrup conserved at 25 and 5°C. The results show that there is no presence of bacteria, yeasts, and molds in the syrups stored at both temperatures during the analysis period. Conclusion: Sheltered from light, the developed syrup of prednisone remained stable at room temperature and/or refrigerator for 30 days.

Keywords: extemporaneous syrup, pediatric drug, prednisone, stability

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686 Early Formation of Adipocere in Subtropical Climate

Authors: Asit K. Sikary, O. P. Murty

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Adipocere formation is a modification of the process of putrefaction. It consists mainly of saturated fatty acids, formed by the post-mortem hydrolysis and hydrogenation of body fats with the help of bacterial enzymes in the presence of warmth, moisture and anaerobic bacteria. In temperate climate, it takes weeks to develop while in India it starts to begin within 4-5 days. In this study, we have collected cases with adipocere formation, which were from the South Delhi region (average room temperature 27-390C) and autopsied at our centre. Details of the circumstances of the death, cause and time of death, surrounding environment and demographic profile of the deceased were taken into account. Total 16 cases were included in this study. Adipocere formation was predominantly present over cheeks, shoulder, breast, flanks, buttocks, and thighs. Out of 16, 11 cases were found in a dry atmosphere, 5 cases were brought from the water. There were 5 cases in which adipocere formation was seen in less than 2 days, and among them, in 1 case, as early as one day. This study showed that adipocere formation can be seen as early as 1 day in a hot and humid environment.

Keywords: adipocere, drowning, hanging, humid environment, strangulation, subtropical climate

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685 Metagenomic analysis of Irish cattle faecal samples using Oxford Nanopore MinION Next Generation Sequencing

Authors: Niamh Higgins, Dawn Howard

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The Irish agri-food sector is of major importance to Ireland’s manufacturing sector and to the Irish economy through employment and the exporting of animal products worldwide. Infectious diseases and parasites have an impact on farm animal health causing profitability and productivity to be affected. For the sustainability of Irish dairy farming, there must be the highest standard of animal health. There can be a lack of information in accounting for > 1% of complete microbial diversity in an environment. There is the tendency of culture-based methods of microbial identification to overestimate the prevalence of species which grow easily on an agar surface. There is a need for new technologies to address these issues to assist with animal health. Metagenomic approaches provide information on both the whole genome and transcriptome present through DNA sequencing of total DNA from environmental samples producing high determination of functional and taxonomic information. Nanopore Next Generation Technologies have the ability to be powerful sequencing technologies. They provide high throughput, low material requirements and produce ultra-long reads, simplifying the experimental process. The aim of this study is to use a metagenomics approach to analyze dairy cattle faecal samples using the Oxford Nanopore MinION Next Generation Sequencer and to establish an in-house pipeline for metagenomic characterization of complex samples. Faecal samples will be obtained from Irish dairy farms, DNA extracted and the MinION will be used for sequencing, followed by bioinformatics analysis. Of particular interest, will be the parasite Buxtonella sulcata, which there has been little research on and which there is no research on its presence on Irish dairy farms. Preliminary results have shown the ability of the MinION to produce hundreds of reads in a relatively short time frame of eight hours. The faecal samples were obtained from 90 dairy cows on a Galway farm. The results from Oxford Nanopore ‘What’s in my pot’ (WIMP) using the Epi2me workflow, show that from a total of 926 classified reads, 87% were from the Kingdom Bacteria, 10% were from the Kingdom Eukaryota, 3% were from the Kingdom Archaea and < 1% were from the Kingdom Viruses. The most prevalent bacteria were those from the Genus Acholeplasma (71 reads), Bacteroides (35 reads), Clostridium (33 reads), Acinetobacter (20 reads). The most prevalent species present were those from the Genus Acholeplasma and included Acholeplasma laidlawii (39 reads) and Acholeplasma brassicae (26 reads). The preliminary results show the ability of the MinION for the identification of microorganisms to species level coming from a complex sample. With ongoing optimization of the pipe-line, the number of classified reads are likely to increase. Metagenomics has the potential in animal health for diagnostics of microorganisms present on farms. This would support wprevention rather than a cure approach as is outlined in the DAFMs National Farmed Animal Health Strategy 2017-2022.

Keywords: animal health, buxtonella sulcata, infectious disease, irish dairy cattle, metagenomics, minION, next generation sequencing

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684 Composition and in Vitro Antimicrobial Activity of Three Eryngium L. Species

Authors: R. Mickiene, A. Friese, U. Rosler, A. Maruska, O. Ragazinskiene

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This research focuses on phytochemistry and antimicrobial activities of compounds isolated and identified from three species of Eryngium. The antimicrobial activity of extracts from Eryngiumplanum L., Eryngium maritimum L., Eryngium campestre L. grown in Lithuania, were tested by the method of series dilutions, against different bacteria species: Escherichia coli, Proteus vulgaris and Staphylococcus aureus with and without antibiotic resistances, originating from livestock. The antimicrobial activity of extracts was described by determination of the minimal inhibitory concentration. Preliminary results show that the minimal inhibitory concentration range between 8.0 % and 17.0 % for the different Eryngium extracts and bacterial species.The total amounts ofphenolic compounds and total amounts of flavonoids were tested in the methanolic extracts of the plants. Identification and evaluation of the phenolic compounds were performed by liquid chromatography. The essential oils were analyzed by gas chromatography mass spectrometry.

Keywords: antimicrobial activities, Eryngium L. species, essential oils, gas chromatography mass spectrometry

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683 Ecotoxicological Safety of Wastewater Treated with Lignocellulosic Adsorbents

Authors: Luísa P. Cruz-Lopes, Artur Figueirinha, Isabel Brás, Bruno Esteves

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Portugal is an important wine and olive oil producer, activities which generate a high quantity of residues commonly called grape stalks and olive cake, respectively. In this work grape stalks and olive cake were used as lignocellulosic adsorbents for wastewater containing lead treatment. To attain a better knowledge of the factors that could influence the quality of the treated wastewater, a chemical characterization of the materials used in the treatment was done. To access the ecotoxicological safety of the treated wastewater, several tests were performed. The results of the toxicity test show that the samples leachate has a mild effect on the living models tested. The tests performed in lemna and bacteria were the most sensible to toxicity effects of the samples. The results obtained in this work evidenced the importance of use of simple and fast toxicity tests to predict impacts in the environment.

Keywords: chemical composition, lignocellulosic residues, ecotoxicological safety, wastewater

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682 The Impact of the Genetic Groups of Microorganisms on the Production of Mousy-Compounds

Authors: Pierre Moulis, Markus Herderich, Doris Rauhut, Patricia Ballestra

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Nowadays, it is starting to be more frequent to detect wines with mousy off-flavor. The reasons behind this could be the significant decrease in sulphur dioxide, the increase in pH, and the trend for spontaneous fermentation in wine. This off-flavor can be produced by Brettanomyces bruxellensis or some Lactic acid bacteria. So far there is no study working on the influence of the genetic group on the production of these microorganisms. Objectives: The objectives of this research are to increase knowledge and to have a better understanding of the microbiological phenomena related to the production of the mousy off-flavor in the wine. Methodologies: In this research, microorganisms were screened in an N-heterocycle assay medium (this medium contained all known precursors) and the production of mousy compounds was quantified by Stir Bar Sorptive Extraction-Gas Chromatography-Mass Spectrometry (SBSE-GC-MS). Main contributions: Brettanomyces bruxellensis and Oenococcus oeni could produce mousiness at a different amount depending on the strain. But there is no group effect.

Keywords: mousy off-flavor, wine, Brettanomyces bruxellensis, Oenococcus oeni

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681 The Effect of Street Dust on Urban Environment

Authors: Turki M. Habeebullah, Abdel Hameed A. A. Awad, Said Munir, Atif M. F. Mohammed, Essam A. Morsy, Abdulaziz R. Seroji

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Street dust has been knoweldged as an important source of air pollution. It does not remain deposited in a place for long, as it is easily resuspended back into the atmosphere. Street dust is a complex mixture derived from different sources: Deposited dust, traffic, tire, and brake wear, construction and demolition processes. The present study aims to evaluate the elementals ”iron, calcium, lead, cadmium, nickel, silicon, and selenium” and microbial “bacteria and fungi” contents associated street dust at the holy mosque areas. The street dust was collected by sweeping an arera~1m2 along the both sides of the road. The particles with diameter ≤ 1.7 µm constitued the highest percentages of the total particulate ≤45 µm. Moreover, The crustal species: iron and calcium were found in the highest concentrations, and proof that demolition and constricution were the main source of street dust. Also, the low biodiversity of microorganisms is attributed to severe weather conditions and characteristics of the arid environment.

Keywords: dust, microbial, environment, street

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680 Antimicrobial Activities of Lactic Acid Bacteria from Fermented Foods and Probiotic Products

Authors: Alec Chabwinja, Cannan Tawonezvi, Jerneja Vidmar, Constance Chingwaru, Walter Chingwaru

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Objective: To evaluate the potential of commercial fermented / probiotic products available in Zimbabwe or internationally, and strains of Lactobacillus plantarum (L. plantarum) as prophylaxis and therapy against diarrhoeal and sexually transmitted infections. Methods: The antimicrobial potential of cultures of lactobacilli enriched from 4 Zimbabwean commercial food/beverage products, namely Dairibord Lacto sour milk (DLSM), Probrand sour milk (PSM), Kefalos Vuka cheese (KVC) and Chibuku opaque beer (COB); three probiotic products obtainable in Europe and internationally; and four strains of L. plantarum obtained from Balkan traditional cheeses and Zimbabwean foods against clinical strains of Escherichia coli (E. coli) and non-clinical strains of Candida albicans and Rhodotorula spp. was assayed using the well diffusion method. Three commercial Agar diffusion assay and a competitive exclusion assay were carried out on Mueller-Hinton agar. Results: Crude cultures of putative lactobacillus strains obtained from Zimbabwean dairy products (Probrand sour milk, Kefalos Vuka vuka cheese and Chibuku opaque beer) exhibited significantly greater antimicrobial activities against clinical strains of E. coli than strains of L. plantarum isolated from Balkan cheeses (CLP1, CLP2 or CLP3) or crude microbial cultures from commercial paediatric probiotic products (BG, PJ and PL) of a culture of Lactobacillus rhamnosus LGG (p < 0.05). Furthermore, the following has high antifungal activities against the two yeasts: supernatant-free microbial pellet (SFMP) from an extract of M. azedarach leaves (27mm ± 2.5) > cell-free culture supernatants (CFCS) from Maaz Dairy sour milk and Mnandi sour milk (approximately 26mm ± 1.8) > CFCS and SFMP from Amansi hodzeko (25mm ± 1.5) > CFCS from Parinari curatellifolia fruit (24mm ± 1.5), SFMP from P. curatellifolia fruit (24mm ± 1.4) and SFMP from mahewu (20mm ± 1.5). These cultures also showed high tolerance to acidic conditions (~pH4). Conclusions: The putative lactobacilli from four commercial Zimbabwean dairy products (Probrand sour milk, Kefalos Vuka vuka cheese and Chibuku opaque beer), and three strains of L. plantarum from Balkan cheeses (CLP1, CLP2 or CLP3) exhibited high antibacterial activities, while Maaz Dairy sour-, Mnandi sour- and Amansi hodzeko milk products had high antifungal activities. Our selection of Zimbabwean probiotic products has potential for further development into probiotic products for use in the control diarrhea caused by pathogenic strains of E. coli or yeast infections. Studies to characterise the probiotic potential of the live cultures in the products are underway.

Keywords: lactic acid bacteria, Staphylococcus aureus, Streptococcus spp, yeast, inhibition, acid tolerance

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679 The Antimicrobrial Effect of Alkaloids (Harmin, Harmalin) Extracted from Peganum harmala (L) Seeds in the South of Algeria (Bousaada)

Authors: Nassima Behidj-Benyounes, Thoraya Dahmene, Nadjiba Chebout

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This work examines the study of the antimicrobrial effect of alkaloids extracted from the seeds of Peganum harmala L (Zygophyllaceae). This natural substance is extracted by using different solvents (aqueous, ethanolic, and hexane). The evaluation of the antimicrobial activity has only dealt with alkaloids. The antimicrobial effect of alkaloids is evaluated on several microorganisms. It has been tested on eight bacterial strains. The extract has been studied by using two yeasts. Finally, three molds have been studied. It should be noted that these agents are characterized by a high frequency of contamination and pathogenicity. Through this study, we note that Staphylococcus aureus, Saccharomyces cerievisae and E. coli are very sensitive in respect of the ethanol extract. Pseudomonas aerogenosa and Penicillium sp. are resistant to this extract. The other microorganisms are moderately sensitive. The study of the antimicrobial activity of different extracts of the Harmel has shown an optimal activity with the ethanol extract.

Keywords: Peganum harmala L., seeds, alkaloids, bacteria, fungi, yeast, antimicrobial activity

Procedia PDF Downloads 380