Search results for: bacterial contamination
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1869

Search results for: bacterial contamination

1569 Progress in Replacing Antibiotics in Farm Animal Production

Authors: Debabrata Biswas

Abstract:

The current trend in the development of antibiotic resistance by multiple bacterial pathogens has resulted in a troubling loss of effective antibiotic options for human. The emergence of multi-drug-resistant pathogens has necessitated higher dosages and combinations of multiple antibiotics, further exacerbating the problem of antibiotic resistance. Zoonotic bacterial pathogens, such as Salmonella, Campylobacter, Shiga toxin-producing Escherichia coli (such as enterohaemorrhagic E. coli or EHEC), and Listeria are the most common and predominant foodborne enteric infectious agents. It was observed that these pathogens gained/developed their ability to survive in the presence of antibiotics either in farm animal gut or farm environment and researchers believe that therapeutic and sub-therapeutic antibiotic use in farm animal production might play an important role in it. The mechanism of action of antimicrobial components used in farm animal production in genomic interplay in the gut and farm environment, has not been fully characterized. Even the risk of promoting the exchange of mobile genetic elements between microbes specifically pathogens needs to be evaluated in depth, to ensure sustainable farm animal production, safety of our food and to mitigate/limit the enteric infection with multiple antibiotic resistant bacterial pathogens. Due to the consumer’s demand and considering the current emerging situation, many countries are in process to withdraw antibiotic use in farm animal production. Before withdrawing use of the sub-therapeutic antibiotic or restricting the use of therapeutic antibiotics in farm animal production, it is essential to find alternative natural antimicrobials for promoting the growth of farm animal and/or treating animal diseases. Further, it is also necessary to consider whether that compound(s) has the potential to trigger the acquisition or loss of genetic materials in zoonotic and any other bacterial pathogens. Development of alternative therapeutic and sub-therapeutic antimicrobials for farm animal production and food processing and preservation and their effective implementation for sustainable strategies for farm animal production as well as the possible risk for horizontal gene transfer in major enteric pathogens will be focus in the study.

Keywords: food safety, natural antimicrobial, sustainable farming, antibiotic resistance

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1568 Interaction of Chemical, Microbiological and Ecological Aspects in the Brown Alga Zonaria Tournefortii

Authors: Sonia Hamiche, Naima Bouzidi, Mohamed Reda Zahi, Yasmina Daghbouche, Abdelmalek Badis, Mohamed El Hattab

Abstract:

This study was carried out on the brown alga Zonaria tourfortii harvested on the central coast of Algeria. The chemical study enabled the characterization of phenolic compounds, mainly acyl phloroglucinol and chromone metabolites. The study isolated a significant quantity of all-cis-5,8,11,14,17 eicosapentanoic acid (EPA). Based on a literature review, we have proposed a biosynthetic pathway leading from EPA to phenolic metabolites. Bacterial screening from the algal surface led to isolate 30 bacterial strains, including 26 Gram+ containing the Staphylococcus and Bacillus genus, and 4 Gram- containing the Acinetobacter and Enterobacteracea genus. In terms of activity profiles, strain S13 (identified as Bacillus amyloliquefaciens based on 16S rRNA technique) proved highly interesting inhibitory activities against target germs, as well as its production of diffusible and volatile compounds. Bacterial cells from the B. amyloliquefaciens S13 strain were used to recover a volatile fraction. Analysis was carried out by gas chromatography-mass spectrometry. The main volatile compounds identified were: 13-epi-manoyl oxide (29.39%), manool (17.39%), 15,16-dinorlabd-8(20)-en-13-one (13.17%), labda-8(17),13Z-dien-15-ol (9. 51%) and 3-acetoxy-13 epimanoyl oxide (5.26%) belonging to the labdane class of diterpenes, the latter having never been described in the category of microbial volatile organic compounds. Ecological aspects were discussed.

Keywords: chemical analysis, acylphloroglucinols, phenolic compounds, microbial volatiles, Zonaria tournefortii

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1567 Clinical and Microbiologic Efficacy and Safety of Imipenem Cilastatin Relebactam in Complicated Infections: A Meta-analysis

Authors: Syeda Sahra, Abdullah Jahangir, Rachelle Hamadi, Ahmad Jahangir, Allison Glaser

Abstract:

Background: Antimicrobial resistance is on the rise. The use of redundant and inappropriate antibiotics is contributing to recurrent infections and resistance. Newer antibiotics with more robust coverage for gram-negative bacteria are in great demand for complicated urinary tract infections (cUTIs), complicated intra-abdominal infections (cIAIs), hospital-acquired bacterial pneumonia (H.A.B.P.), and ventilator-associated bacterial pneumonia (V.A.B.P.). Objective: We performed this meta-analysis to evaluate the efficacy and safety profile of a new antibiotic, Imipenem/cilastatin/relebactam, compared to other broad-spectrum antibiotics for complicated infections. Search Strategy: We conducted a systemic review search on PubMed, Embase, and Central Cochrane Registry. Selection Criteria: We included randomized clinical trials (R.C.T.s) with the standard of care as comparator arm with Imipenem/cilastatin/relebactam as intervention arm. Analysis: For continuous variables, the mean difference was used. For discrete variables, we used the odds ratio. For effect sizes, we used a confidence interval of 95%. A p-value of less than 0.05 was used for statistical significance. Analysis was done using a random-effects model irrespective of heterogeneity. Heterogeneity was evaluated using the I2 statistic. Results: The authors observed similar efficacy at clinical and microbiologic response levels on early follow-up and late follow-up compared to the established standard of care. The incidence of drug-related adverse events, serious adverse events, and drug discontinuation due to adverse events were comparable across both groups. Conclusion: Imipenem/cilastatin/relebactam has a non-inferior safety and efficacy profile compared to peer antibiotics to treat severe bacterial infections (cUTIs, cIAIs, H.A.B.P., V.A.B.P.).

Keywords: bacterial pneumonia, complicated intra-abdominal infections, complicated urinary tract infection, Imipenem, cilastatin, relebactam

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1566 Application and Limitation of Heavy Metal Pollution Indicators in Coastal Environment of Pakistan

Authors: Noor Us Saher

Abstract:

Oceans and Marine areas have a great importance, mainly regarding food resources, fishery products and reliance of livelihood. Aquatic pollution is common due to the incorporation of various chemicals mainly entering from urbanization, industrial and commercial facilities, such as oil and chemical spills. Many hazardous wastes and industrial effluents contaminate the nearby areas and initiate to affect the marine environment. These contaminated conditions may become worse in those aquatic environments situated besides the world’s largest cities, which are hubs of various commercial activities. Heavy metal contamination is one of the most important predicaments for marine environments and during past decades this problem has intensified due to an increase in urbanization and industrialization. Coastal regions of Pakistan are facing severe threats from various organic and inorganic pollutants, especially the estuarine and coastal areas of Karachi city, the most populated and industrialized city situated along the coastline. Metal contamination causes severe toxicity in biota resulting the degradation of Marine environments and depletion of fishery resources and sustainability. There are several abiotic (air, water and sediment) and biotic (fauna and flora) indicators that indicate metal contamination. However, all these indicators have certain limitations and complexities, which delay their implementation for rehabilitation and conservation in the marine environment. The inadequate evidences have presented on this significant topic till the time and this study discussed metal pollution and its consequences along the marine environment of Pakistan. This study further helps in identification of possible hazards for the ecological system and allied resources for management strategies and decision making for sustainable approaches.

Keywords: coastal and estuarine environment, heavy metals pollution, pollution indicators, Pakistan

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1565 Evaluation of Immunostimulant Potential of Proteoliposomes Derived from Vibrio anguillarum Administered by Immersion in Zebrafish (Danio rerio)

Authors: M. Caruffo, P. Navarrete, C. G. Feijoo, L. Sáenz

Abstract:

Disease prevention through the use of vaccines has been crucial to achieve the current level of production in the salmon industry. However, vaccines have been developed based largely on inactivated bacterial formulations, using the whole pathogen. These formulations have demonstrated excellent efficacy against extracellular bacterial pathogens. However diseases with the greatest economic impacts correspond to intracellular bacterial and viral pathogens, vaccines based on these types of agents have shown a discrete effectiveness. It is for these reasons that the development of subunit vaccines based on defined antigens offers a promising solution. The main problem is that subunit vaccines offer a low immunogenicity, since they lack immunostimulatory elements, so that the development of new adjuvants platforms becomes an important challenge for this type of formulations. We evaluate the effect of a formulation based on proteoliposomes of Vibrio anguillarum administered by immersion as a new adjuvant strategy, allowing efficient stimulation of the innate immune system. Proteoliposomes physicochemical properties were evaluated in its ability to produce an inflammatory process. Using zebrafish (Danio rerio) larvae as a model species and the transgenic line (Tg(mpx: GFP)i114) allowed us to track the neutrophil migration in real time. Additionally we evaluated the gene expression of some molecular markers involved in the development of the innate immune response characterizing the adjuvant capacity of the formulation.

Keywords: adjuvants, vaccine development, zebrafish, innate immunity

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1564 Development of an Appropriate Method for the Determination of Multiple Mycotoxins in Pork Processing Products by UHPLC-TCFLD

Authors: Jason Gica, Yi-Hsieng Samuel Wu, Deng-Jye Yang, Yi-Chen Chen

Abstract:

Mycotoxins, harmful secondary metabolites produced by certain fungi species, pose significant risks to animals and humans worldwide. Their stable properties lead to contamination during grain harvesting, transportation, and storage, as well as in processed food products. The prevalence of mycotoxin contamination has attracted significant attention due to its adverse impact on food safety and global trade. The secondary contamination pathway from animal products has been identified as an important route of exposure, posing health risks for livestock and humans consuming contaminated products. Pork, one of the highly consumed meat products in Taiwan according to the National Food Consumption Database, plays a critical role in the nation's diet and economy. Given its substantial consumption, pork processing products are a significant component of the food supply chain and a potential source of mycotoxin contamination. This study is paramount for formulating effective regulations and strategies to mitigate mycotoxin-related risks in the food supply chain. By establishing a reliable analytical method, this research contributes to safeguarding public health and enhancing the quality of pork processing products. The findings will serve as valuable guidance for policymakers, food industries, and consumers to ensure a safer food supply chain in the face of emerging mycotoxin challenges. An innovative and efficient analytical approach is proposed using Ultra-High Performance Liquid Chromatography coupled with Temperature Control Fluorescence Detector Light (UHPLC-TCFLD) to determine multiple mycotoxins in pork meat samples due to its exceptional capacity to detect multiple mycotoxins at the lowest levels of concentration, making it highly sensitive and reliable for comprehensive mycotoxin analysis. Additionally, its ability to simultaneously detect multiple mycotoxins in a single run significantly reduces the time and resources required for analysis, making it a cost-effective solution for monitoring mycotoxin contamination in pork processing products. The research aims to optimize the efficient mycotoxin QuEChERs extraction method and rigorously validate its accuracy and precision. The results will provide crucial insights into mycotoxin levels in pork processing products.

Keywords: multiple-mycotoxin analysis, pork processing products, QuEChERs, UHPLC-TCFLD, validation

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1563 Bacteria Immobilized Electrospun Fibrous Biocomposites for Cr (VI) Remediation in Water

Authors: Omer Faruk Sarioglu, Asli Celebioglu, Turgay Tekinay, Tamer Uyar

Abstract:

Fibrous biocomposites were developed by immobilization of a Cr(VI) reducing bacterial strain, morganella morganii STB5, on electrospun polystyrene (PS) and polysulfone (PSU) webs. Cr(VI) removal characteristics of STB5/PS and STB5/PSU fibrous biocomposites were determined at 25 mg L-1 of initial Cr(VI) and 70.41% and 68.27% of removal were observed within 72 h, respectively. Reusability test results indicate that both biocomposites are potentially reusable and can be used for at least 5 cycles. After storage test results suggest that the biocomposites can be stored awhile without losing their Cr(VI) bioremoval capabilities. SEM images of STB5 immobilized PS and PSU webs after the reusability test exhibit strong attachment of bacterial biofilms onto fibrous surfaces. Our results are quite promising and suggesting that reusable bacteria immobilized electrospun fibrous biocomposites might be applicable for Cr(VI) remediation in water systems.

Keywords: electrospinning, polystyrene, polysulfone, Cr(VI) bioremoval, environmental sustainability

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1562 The Staphylococcus aureus Exotoxin Recognition Using Nanobiosensor Designed by an Antibody-Attached Nanosilica Method

Authors: Hamed Ahari, Behrouz Akbari Adreghani, Vadood Razavilar, Amirali Anvar, Sima Moradi, Hourieh Shalchi

Abstract:

Considering the ever increasing population and industrialization of the developmental trend of humankind's life, we are no longer able to detect the toxins produced in food products using the traditional techniques. This is due to the fact that the isolation time for food products is not cost-effective and even in most of the cases, the precision in the practical techniques like the bacterial cultivation and other techniques suffer from operator errors or the errors of the mixtures used. Hence with the advent of nanotechnology, the design of selective and smart sensors is one of the greatest industrial revelations of the quality control of food products that in few minutes time, and with a very high precision can identify the volume and toxicity of the bacteria. Methods and Materials: In this technique, based on the bacterial antibody connection to nanoparticle, a sensor was used. In this part of the research, as the basis for absorption for the recognition of bacterial toxin, medium sized silica nanoparticles of 10 nanometer in form of solid powder were utilized with Notrino brand. Then the suspension produced from agent-linked nanosilica which was connected to bacterial antibody was positioned near the samples of distilled water, which were contaminated with Staphylococcus aureus bacterial toxin with the density of 10-3, so that in case any toxin exists in the sample, a connection between toxin antigen and antibody would be formed. Finally, the light absorption related to the connection of antigen to the particle attached antibody was measured using spectrophotometry. The gene of 23S rRNA that is conserved in all Staphylococcus spp., also used as control. The accuracy of the test was monitored by using serial dilution (l0-6) of overnight cell culture of Staphylococcus spp., bacteria (OD600: 0.02 = 107 cell). It showed that the sensitivity of PCR is 10 bacteria per ml of cells within few hours. Result: The results indicate that the sensor detects up to 10-4 density. Additionally, the sensitivity of the sensors was examined after 60 days, the sensor by the 56 days had confirmatory results and started to decrease after those time periods. Conclusions: Comparing practical nano biosensory to conventional methods like that culture and biotechnology methods(such as polymerase chain reaction) is accuracy, sensitiveness and being unique. In the other way, they reduce the time from the hours to the 30 minutes.

Keywords: exotoxin, nanobiosensor, recognition, Staphylococcus aureus

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1561 Exploring the Prebiotic Potential of Glucosamine

Authors: Shilpi Malik, Ramneek Kaur, Archita Gupta, Deepshikha Yadav, Ashwani Mathur, Manisha Singh

Abstract:

Glucosamine (GS) is the most abundant naturally occurring amino monosaccharide and is normally produced in human body via cellular glucose metabolism. It is regarded as the building block of cartilage matrix and is also an essential component of cartilage matrix repair mechanism. Besides that, it can also be explored for its prebiotic potential as many bacterial species are known to utilize the amino sugar by acquiring them to form peptidoglycans and lipopolysaccharides in the bacterial cell wall. Glucosamine can therefore be considered for its fermentation by bacterial species present in the gut. Current study is focused on exploring the potential of glucosamine as prebiotic. The studies were done to optimize considerable concentration of GS to reach GI tract and being fermented by the complex gut microbiota and food grade GS was added to various Simulated Fluids of Gastro-Intestinal Tract (GIT) such as Simulated Saliva, Gastric Fluid (Fast and Fed State), Colonic fluid, etc. to detect its degradation. Since it was showing increase in microbial growth (CFU) with time, GS was Further, encapsulated to increase its residential time in the gut, which exhibited improved resistance to the simulated Gut conditions. Moreover, prepared microspehres were optimized and characterized for their encapsulation efficiency and toxicity. To further substantiate the prebiotic activity of Glucosamine, studies were also performed to determine the effect of Glucosamine on the known probiotic bacterial species, i.e. Lactobacillus delbrueckii (MTCC 911) and Bifidobacteriumbifidum (MTCC 5398). Culture conditions for glucosamine will be added in MRS media in anaerobic tube at 0.20%, 0.40%, 0.60%, 0.80%, and 1.0%, respectively. MRS media without GS was included in this experiment as the control. All samples were autoclaved at 118° C for 15 min. Active culture was added at 5% (v/v) to each anaerobic tube after cooling to room temperature and incubated at 37° C then determined biomass and pH and viable count at incubation 18h. The experiment was completed in triplicate and the results were presented as Mean ± SE (Standard error).The experimental results are conclusive and suggest Glucosamine to hold prebiotic properties.

Keywords: gastro intestinal tract, microspheres, peptidoglycans, simulated fluid

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1560 Effects of Environmental Parameters on Salmonella Contaminated in Harvested Oysters (Crassostrea lugubris and Crassostrea belcheri)

Authors: Varangkana Thaotumpitak, Jarukorn Sripradite, Saharuetai Jeamsripong

Abstract:

Environmental contamination from wastewater discharges originated from anthropogenic activities introduces the accumulation of enteropathogenic bacteria in aquatic animals, especially in oysters, and in shellfish harvesting areas. The consumption of raw or partially cooked oysters can be a risk for seafood-borne diseases in human. This study aimed to evaluate the relationship between the presence of Salmonella in oyster meat samples, and environmental factors (ambient air temperature, relative humidity, gust wind speed, average wind speed, tidal condition, precipitation and season) by using the principal component analysis (PCA). One hundred and forty-four oyster meat samples were collected from four oyster harvesting areas in Phang Nga province, Thailand from March 2016 to February 2017. The prevalence of Salmonella of each site was ranged from 25.0-36.11% in oyster meat. The results of PCA showed that ambient air temperature, relative humidity, and precipitation were main factors correlated with Salmonella detection in these oysters. Positive relationship was observed between positive Salmonella in the oysters and relative humidity (PC1=0.413) and precipitation (PC1=0.607), while the negative association was found between ambient air temperature (PC1=0.338) and the presence of Salmonella in oyster samples. These results suggested that lower temperature and higher precipitation and higher relative humidity will possibly effect on Salmonella contamination of oyster meat. During the high risk period, harvesting of oysters should be prohibited to reduce pathogenic bacteria contamination and to minimize a hazard of humans from Salmonellosis.

Keywords: oyster, Phang Nga Bay, principal component analysis, Salmonella

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1559 Effect of Entomopathogenic Fungi on the Food Consumption of Acrididae Species

Authors: S. Kumar, R. Sultana

Abstract:

This study was conducted to evaluate the effect of Aspergillus species on acridid populations which are major agricultural pests of rice, sugarcane, wheat, maize and fodder crops in Pakistan. Three and replicates i.e. Aspergillus flavus, A. fumigatus and A. niger, excluding the control, were held under laboratory conditions. It was observed that consumption faecal production of acridids was significantly reduced after the pathogenic application of Aspergillus. In the control replicate, the mortality ratio for stage (N4-N6) was maximum on day 2nd i.e. [F10.7 = 18.33, P < 0.05] followed by [F4.20 = 07.85, P < 0.05] and [F3.77 = 06.11, P < 0.05] on 4th and 3rd day, respectively. Similarly, it was a minimum i.e. [F0.48 = 84.65, P < 0.05] on the 1st day. It was also noted that faecal production of Acridid nymphs was not significantly affected when treated with conidial concentration in H2O formulation; however, it was significantly reduced after the contamination with conidial concentration in oil. The high morality of acridids after contamination of Aspergillus supports their use as bio-control agent for reducing pest population. The present study recommends that exploration and screening must be conducted to provide additional pathogens for evaluation as potential biological control against grasshoppers and locusts.

Keywords: acridid, agriculture, formulation, grasshoppers

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1558 Discrimination of Bio-Analytes by Using Two-Dimensional Nano Sensor Array

Authors: P. Behera, K. K. Singh, D. K. Saini, M. De

Abstract:

Implementation of 2D materials in the detection of bio analytes is highly advantageous in the field of sensing because of its high surface to volume ratio. We have designed our sensor array with different cationic two-dimensional MoS₂, where surface modification was achieved by cationic thiol ligands with different functionality. Green fluorescent protein (GFP) was chosen as signal transducers for its biocompatibility and anionic nature, which can bind to the cationic MoS₂ surface easily, followed by fluorescence quenching. The addition of bio-analyte to the sensor can decomplex the cationic MoS₂ and GFP conjugates, followed by the regeneration of GFP fluorescence. The fluorescence response pattern belongs to various analytes collected and transformed to linear discriminant analysis (LDA) for classification. At first, 15 different proteins having wide range of molecular weight and isoelectric points were successfully discriminated at 50 nM with detection limit of 1 nM. The sensor system was also executed in biofluids such as serum, where 10 different proteins at 2.5 μM were well separated. After successful discrimination of protein analytes, the sensor array was implemented for bacteria sensing. Six different bacteria were successfully classified at OD = 0.05 with a detection limit corresponding to OD = 0.005. The optimized sensor array was able to classify uropathogens from non-uropathogens in urine medium. Further, the technique was applied for discrimination of bacteria possessing resistance to different types and amounts of drugs. We found out the mechanism of sensing through optical and electrodynamic studies, which indicates the interaction between bacteria with the sensor system was mainly due to electrostatic force of interactions, but the separation of native bacteria from their drug resistant variant was due to Van der Waals forces. There are two ways bacteria can be detected, i.e., through bacterial cells and lysates. The bacterial lysates contain intracellular information and also safe to analysis as it does not contain live cells. Lysates of different drug resistant bacteria were patterned effectively from the native strain. From unknown sample analysis, we found that discrimination of bacterial cells is more sensitive than that of lysates. But the analyst can prefer bacterial lysates over live cells for safer analysis.

Keywords: array-based sensing, drug resistant bacteria, linear discriminant analysis, two-dimensional MoS₂

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1557 Characterization of Bacteriophage for Biocontrol of Pseudomonas syringae, Causative Agent of Canker in Prunus spp.

Authors: Mojgan Rabiey, Shyamali Roy, Billy Quilty, Ryan Creeth, George Sundin, Robert W. Jackson

Abstract:

Bacterial canker is a major disease of Prunus species such as cherry (Prunus avium). It is caused by Pseudomonas syringae species including P. syringae pv. syringae (Pss) and P. syringae pv. morsprunorum race 1 (Psm1) and race 2 (Psm2). Concerns over the environmental impact of, and developing resistance to, copper controls call for alternative approaches to disease management. One method of control could be achieved using naturally occurring bacteriophage (phage) infective to the bacterial pathogens. Phages were isolated from soil, leaf, and bark of cherry trees in five locations in the South East of England. The phages were assessed for their host range against strains of Pss, Psm1, and Psm2. The phages exhibited a differential ability to infect and lyse different Pss and Psm isolates as well as some other P. syringae pathovars. However, the phages were unable to infect beneficial bacteria such as Pseudomonas fluorescens. A subset of 18 of these phages were further characterised genetically (Random Amplification of Polymorphic DNA-PCR fingerprinting and sequencing) and using electron microscopy. The phages are tentatively identified as belonging to the order Caudovirales and the families Myoviridae, Podoviridae, and Siphoviridae, with genetic material being dsDNA. Future research will fully sequence the phage genomes. The efficacy of the phage, both individually and in cocktails, to reduce disease progression in vivo will be investigated to understand the potential for practical use of these phages as biocontrol agents.

Keywords: bacteriophage, pseudomonas, bacterial cancker, biological control

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1556 Landscape Management in the Emergency Hazard Planning Zone of the Nuclear Power Plant Temelin: Preventive Improvement of Landscape Functions

Authors: Ivana Kašparová, Emilie Pecharová

Abstract:

The experience of radiological contamination of land, especially after the Chernobyl and Fukushima disasters have shown the need to explore possibilities to the capture of radionuclides in the area affected and to adapt the landscape management to this purpose ex –ante the considered accident in terms of prevention. The project‚ Minimizing the impact of radiation contamination on land in the emergency zone of Temelin NPP‘ (2012-2015), dealt with the possibility of utilization of wetlands as retention sites for water carrying radionuclides in the case of a radiation accident. A model artificial wetland was designed and adopted as a utility model by the Ministry of Industry and Trade of the Czech Republic. The article shows the conditions of construction of designed wetlands in the landscape with regard to minimizing the negative effect on agricultural production and enhancing the hydrological functionality of the landscape.

Keywords: artificial wetland, land use/ land cover, old maps, surface-to-water transport of radionuclides

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1555 Sustainable Agriculture Practices Using Bacterial-mediated Alleviation of Salinity Stress in Crop Plants

Authors: Mohamed Trigui, Fatma Masmoudi, Imen Zouari

Abstract:

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

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

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1554 A Script for Presentation to the Management of a Teaching Hospital on MYCIN: A Clinical Decision Support System

Authors: Rashida Suleiman, Asamoah Jnr. Boakye, Suleiman Ahmed Ibn Ahmed

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In recent years, there has been an enormous success in discoveries of scientific knowledge in medicine coupled with the advancement of technology. Despite all these successes, diagnoses and treatment of diseases have become complex. MYCIN is a groundbreaking illustration of a clinical decision support system (CDSS), which was developed to assist physicians in the diagnosis and treatment of bacterial infections by providing suggestions for antibiotic regimens. MYCIN was one of the earliest expert systems to demonstrate how CDSSs may assist human decision-making in complicated areas. Relevant databases were searched using google scholar, PubMed and general Google search, which were peculiar to clinical decision support systems. The articles were then screened for a comprehensive overview of the functionality, consultative style and statistical usage of MYCIN, a clinical decision support system. Inferences drawn from the articles showed some usage of MYCIN for problem-based learning among clinicians and students in some countries. Furthermore, the data demonstrated that MYCIN had completed clinical testing at Stanford University Hospital following years of research. The system (MYCIN) was shown to be extremely accurate and effective in diagnosing and treating bacterial infections, and it demonstrated how CDSSs might enhance clinical decision-making in difficult circumstances. Despite the challenges MYCIN presents, the benefits of its usage to clinicians, students and software developers are enormous.

Keywords: clinical decision support system, MYCIN, diagnosis, bacterial infections, support systems

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1553 Functional Diversity of Pseudomonas: Role in Stimulation of Bean Germination and Common Blight Biocontrol

Authors: Slimane Mokrani, Nabti El hafid

Abstract:

Description of the subject: Currently, several efforts focus on the study of biodiversity, microbial biotechnology, and the use of ecological strategies. Objectives: The aim of this present work is to determine the functional diversity of bacteria in rhizospheric and non-rhizospheric soils of different plants. Methods: Bacteria were isolated from soil and identified based on physiological and biochemical characters and genotypic taxonomy performed by 16S rDNA and BOX-PCR. As well as the characterization of various PGPR traits. Then, they are tested for their effects on the stimulation of seed germination and the growth of Phaseolus vulgaris L. As well as their biological control activities with regard to the phytopathogenic bacterial isolate Xapf. Results and Discussion: The biochemical and physiological identification of 75 bacterial isolates made it possible to associate them with the two groups of fluorescent Pseudomonas (74.67%) and non-fluorescent Pseudomonas (25.33%). The identification by 16S rDNA of 27 strains made it possible to attribute the majority of the strains to the genus Pseudomonas (81.48%), Serratia (7.41%) and Bacillus (11.11%). The bacterial strains showed a high capacity to produce IAA, siderophores, HCN and to solubilize phosphate. A significant stimulation of germination and growth was observed by applying the Pseudomonas strains. Furthermore, significant reductions in the severity and intensity of the disease caused caused by Xapf were observed. Conclusion: The bacteria described in this present study endowed with different PGPR activities seem to be very promising for their uses as biological control agents and bio-fertilization.

Keywords: biofertilization, biological control, phaseolus vulgaris L, pseudomonas, Xanthomonas axonopodis pv. phaseoli var. fuscans and common blight

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1552 Isolation of Biosurfactant Producing Spore-Forming Bacteria from Oman: Potential Applications in Bioremediation

Authors: Saif N. Al-Bahry, Yahya M. Al-Wahaibi, Abdulkadir E. Elshafie, Ali S. Al-Bemani, Sanket J. Joshi

Abstract:

Environmental pollution is a global problem and best possible solution is identifying and utilizing native microorganisms. One possible application of microbial product -biosurfactant is in bioremediation of hydrocarbon contaminated sites. We have screened forty two different petroleum contaminated sites from Oman, for biosurfactant producing spore-forming bacterial isolates. Initial screening showed that out of 42 soil samples, three showed reduction in surface tension (ST) and interfacial tension (IFT) within 24h of incubation at 40°C. Out of those 3 soil samples, one was further selected for isolation of bacteria and 14 different bacteria were isolated in pure form. Of those 14 spore-forming, rod shaped bacteria, two showed highest reduction in ST and IFT in the range of 70mN/m to < 35mN/m and 26.69mN/m to < 9mN/m, respectively within 24h. These bacterial biosurfactants may be utilized for bioremediation of oil-spills.

Keywords: bioremediation, hydrocarbon pollution, spore-forming bacteria, bio-surfactant

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1551 Quorum Quenching Activities of Bacteria Isolated from Red Sea Sediments

Authors: Zahid Rehman, TorOve Leiknes

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Quorum sensing (QS) is the process by which bacteria communicate with each other through small signaling molecules, such as N-acylhomoserine lactones (AHLs). Also, certain bacteria have the ability to degrade AHL molecules by a process referred to as quorum quenching (QQ); therefore, QQ can be used to control bacterial infections and biofilm formation. In this study, we aimed to identify new species of bacteria with QQ activities. To achieve this, sediments from Red Sea were collected either in the close vicinity of Sea grass or from area with no vegetation. From these samples, we isolated 72 bacterial strains and tested their ability to degrade/inactivate AHL molecules. Chromobacterium violaceum based bioassay was used in initial screening of isolates for QQ activity. The QQ activity of the positive isolates was further confirmed and quantified by employing liquid chromatography and mass spectrometry. These analyses showed that isolated bacterial strain could degrade AHL molecules with different acyl chain length and modifications. Sequencing of 16S-rRNA genes of positive isolates revealed that they belong to three different genera. Specifically, two isolates belong to genus Erythrobacter, four to Labrenzia and one isolate belongs to Bacterioplanes. Time course experiment showed that isolate belonging to genus Erythrobacter could degrade AHLs faster than other isolates. Furthermore, these isolates were tested for their ability to inhibit formation of biofilm and degradation of 3OXO-C12 AHLs produced by P. aeruginosa PAO1. Our results showed that isolate VG12 is better at controlling biofilm formation. This aligns with the ability of VG12 to cause at least 10-fold reduction in the amount of different AHLs tested.

Keywords: quorum sensing, biofilm, quorum quenching, anti-biofouling

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1550 Mycoflora and Aflatoxin Contamination of Kokoro: A Nigerian Maize Snack

Authors: D. A. Onifade

Abstract:

Kokoro is maize snack which is very popular among poor masses in Nigeria who consume it along with gari(a cassava product) as lunch on a regular basis. In this study, fungal contaminants of kokoro were characterized and its aflatoxin content determined. A total of 30 fungal isolates were obtained from kokoro samples and they belong to 3 different species. Aspergillus flavus had the highest frequency of occurrence of 73.33% while Penicillium species had the lowest (6.66%). Different concentration of aflatoxin B1 was detected in some of the kokoro samples analyzed. Sample D had the highest concentration of 7.25 parts per billion (ppb). The lowest concentration detected was 0.06 ppb in sample P. No aflatoxin G1 and G2 was detected in all the kokoro samples with exception of sample P which contained 2.54 ppb aflatoxin G1.According to international standards some of the kokoro samples are not suitable for human consumption because of high-level aflatoxin which was above the recommended level. Therefore, production of kokoro should be standardized and appropriate packaging materials utilized to prevent the growth of aflatoxigenic fungi. This is to safeguard the health of many poor Nigerians who consume it on a regular basis.

Keywords: kokoro, maize snack, aflatoxin, contamination, mould, Nigeria

Procedia PDF Downloads 298
1549 Hybrid Molecules: A Promising Approach to Design Potent Antimicrobial and Anticancer Drugs

Authors: Blessing Atim Aderibigbe

Abstract:

A series of amine/ester-linked hybrid compounds containing pharmacophores, such as ursolic acid, oleanolic acid, ferrocene and bisphosphonates, were synthesized in an attempt to develop potent antibacterial and anticancer agents. Their structures were analyzed and confirmed using Nuclear Magnetic Resonance, Fourier Transform Infrared Spectroscopy, and mass spectroscopy. All the synthesized hybrid compounds were evaluated for their antibacterial activities against eleven selected bacterial strains using a serial dilution method. Some of the compounds displayed significant antibacterial activity against most of the bacterial and fungal strains. In addition, the in vitro cytotoxicity of these compounds was also performed against selected cancer cell lines. Some of the compounds were also found to be more active than their parent compounds, revealing the efficacy of designing hybrid molecules using plant-based bioactive agents.

Keywords: ursolic acid, hybrid drugs, oleanolic acid, bisphosphonates

Procedia PDF Downloads 55
1548 Network Analysis to Reveal Microbial Community Dynamics in the Coral Reef Ocean

Authors: Keigo Ide, Toru Maruyama, Michihiro Ito, Hiroyuki Fujimura, Yoshikatu Nakano, Shoichiro Suda, Sachiyo Aburatani, Haruko Takeyama

Abstract:

Understanding environmental system is one of the important tasks. In recent years, conservation of coral environments has been focused for biodiversity issues. The damage of coral reef under environmental impacts has been observed worldwide. However, the casual relationship between damage of coral and environmental impacts has not been clearly understood. On the other hand, structure/diversity of marine bacterial community may be relatively robust under the certain strength of environmental impact. To evaluate the coral environment conditions, it is necessary to investigate relationship between marine bacterial composition in coral reef and environmental factors. In this study, the Time Scale Network Analysis was developed and applied to analyze the marine environmental data for investigating the relationship among coral, bacterial community compositions and environmental factors. Seawater samples were collected fifteen times from November 2014 to May 2016 at two locations, Ishikawabaru and South of Sesoko in Sesoko Island, Okinawa. The physicochemical factors such as temperature, photosynthetic active radiation, dissolved oxygen, turbidity, pH, salinity, chlorophyll, dissolved organic matter and depth were measured at the coral reef area. Metagenome and metatranscriptome in seawater of coral reef were analyzed as the biological factors. Metagenome data was used to clarify marine bacterial community composition. In addition, functional gene composition was estimated from metatranscriptome. For speculating the relationships between physicochemical and biological factors, cross-correlation analysis was applied to time scale data. Even though cross-correlation coefficients usually include the time precedence information, it also included indirect interactions between the variables. To elucidate the direct regulations between both factors, partial correlation coefficients were combined with cross correlation. This analysis was performed against all parameters such as the bacterial composition, the functional gene composition and the physicochemical factors. As the results, time scale network analysis revealed the direct regulation of seawater temperature by photosynthetic active radiation. In addition, concentration of dissolved oxygen regulated the value of chlorophyll. Some reasonable regulatory relationships between environmental factors indicate some part of mechanisms in coral reef area.

Keywords: coral environment, marine microbiology, network analysis, omics data analysis

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1547 Bacterial Community Diversity in Soil under Two Tillage Systems

Authors: Dalia Ambrazaitienė, Monika Vilkienė, Danute Karcauskienė, Gintaras Siaudinis

Abstract:

The soil is a complex ecosystem that is part of our biosphere. The ability of soil to provide ecosystem services is dependent on microbial diversity. T Tillage is one of the major factors that affect soil properties. The no-till systems or shallow ploughless tillage are opposite of traditional deep ploughing, no-tillage systems, for instance, increase soil organic matter by reducing mineralization rates and stimulating litter concentrations of the top soil layer, whereas deep ploughing increases the biological activity of arable soil layer and reduces the incidence of weeds. The role of soil organisms is central to soil processes. Although the number of microbial species in soil is still being debated, the metagenomic approach to estimate microbial diversity predicted about 2000 – 18 000 bacterial genomes in 1 g of soil. Despite the key role of bacteria in soil processes, there is still lack of information about the bacterial diversity of soils as affected by tillage practices. This study focused on metagenomic analysis of bacterial diversity in long-term experimental plots of Dystric Epihypogleyic Albeluvisols in western part of Lithuania. The experiment was set up in 2013 and had a split-plot design where the whole-plot treatments were laid out in a randomized design with three replicates. The whole-plot treatments consisted of two tillage methods - deep ploughing (22-25 cm) (DP), ploughless tillage (7-10 cm) (PT). Three subsamples (0-20 cm) were collected on October 22, 2015 for each of the three replicates. Subsamples from the DP and PT systems were pooled together wise to make two composition samples, one representing deep ploughing (DP) and the other ploughless tillage (PT). Genomic DNA from soil sample was extracted from approximately 200 mg field-moist soil by using the D6005 Fungal/Bacterial Miniprep set (Zymo Research®) following the manufacturer’s instructions. To determine bacterial diversity and community composition, we employed a culture – independent approach of high-throughput pyrosequencing of the 16S rRNA gene. Metagenomic sequencing was made with Illumina MiSeq platform in Base Clear Company. The microbial component of soil plays a crucial role in cycling of nutrients in biosphere. Our study was a preliminary attempt at observing bacterial diversity in soil under two common but contrasting tillage practices. The number of sequenced reads obtained for PT (161 917) was higher than DP (131 194). The 10 most abundant genus in soil sample were the same (Arthrobacter, Candidatus Saccharibacteria, Actinobacteria, Acidobacterium, Mycobacterium, Bacillus, Alphaproteobacteria, Longilinea, Gemmatimonas, Solirubrobacter), just the percent of community part was different. In DP the Arthrobacter and Acidobacterium consist respectively 8.4 % and 2.5%, meanwhile in PT just 5.8% and 2.1% of all community. The Nocardioides and Terrabacter were observed just in PT. This work was supported by the project VP1-3.1-ŠMM-01-V-03-001 NKPDOKT and National Science Program: The effect of long-term, different-intensity management of resources on the soils of different genesis and on other components of the agro-ecosystems [grant number SIT-9/2015] funded by the Research Council of Lithuania.

Keywords: deep ploughing, metagenomics, ploughless tillage, soil community analysis

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1546 Biohydrogen Production from Rice Water Using Bacteria Isolated from Wetland Sediment

Authors: Jerry John T. M., Sylas V. P., Shijo Joy

Abstract:

Hydrogen is the most essential gas that can be used for many purposes. During the production of hydrogen using raw materials like Soil and leftover cooked rice water (kanjivellam), the major by-product formed is water. Soil is collected from three different places in kottayam district: Kallara, Meenachilar, and Athirampuzha. Collected samples are mixed with rice water and tested for traces of hydrogen using a biohydrogen sensor after 72 hours. The result was the presence of hydrogen in all the 3 samples. After streaking, PCR and gel electrophoresis detected the bacteria which produced the hydrogen. RGCB Thiruvananthapuram conducted the sequencing of the PCR resultant. And identified the bacterial strains. Five variants of Bacillus bacteria ( (1) Bacillus cereus strain JTM GenBank: OP278839.1 (2) Bacillus toyonensis strain JTM2 GenBank: OP278841.1 (3) Bacillus anthracis strain JTM_SR2989-3-R_H08 GenBank: OP278960.1 (4) Bacillus thuringiensis strain JRY1 GenBank: OP278976.1 (5) Bacillus anthracis strain JTM_SR2989-3-F_H07 GenBank: OP278959.1 ) are identified and successfully registered in NCBI Gen bank. These Bacillus bacteria are major types of Rhizobacteria that can form spores and can survive in the soil for a long time period under harsh environmental conditions. Also, plant growth is enhanced by PGPR (Plant growth promoting rhizobacteria) through the induction of systemic resistance, antibiosis, and competitive omission. The molecular sequencing was submitted to the NCBI Gen Bank, and the accession numbers were allotted for the bacterial cultures.

Keywords: bio hydrogen production, bacterial bio hydrogen production, plant related to bacillus bacteria., bacillus bacteria study

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1545 Prevalence and Evaluation of Antimicrobial Activity of Dodonaea viscosa Extract and Antibacterial Agents against Salmonella spp. Isolated from Poultry

Authors: Shayma Munqith Al-Baker, Fadhl Ahmed Saeed Al-Gasha’a, Samira Hamid Hanash, Ahmed Ali Al-Hazmi

Abstract:

A total of 200 samples (180 fecal materials and 20 organ samples) were collected from (5 different poultry farms, 10 local poultry shops, 5 houses poultry, 5 Eggs stores shops and 5 hand slaughters centers) in Ibb city, Yemen, 2014. According to morphological, cultural, as well as biochemical characterization and serological tests, 59 29.5% isolates were identified as Salmonella spp. and all Salmonella isolates were categorized by serotype, which comprised of, 37 62.71% Salmonella Typhimurium serovar, 21 35.59%. Salmonella Enteritidis serovar and 11.69% Salmonella Heidelberg serovar. Antibiotic sensitivity test was done for bacterial isolates and the results showed there were clear differences in antibiotic resistant. Antimicrobial susceptibility of the isolates varies as follows: Ofloxacin 79.66%, Ciprofloxacin 67.80%, Colistin 59.32% and Gentamycin 52.54%. All of isolates were resistant to Erythromycin, Penicillin and Lincomycin. Antibacterial activity was done for both aqueous and ethanol extracts of Dodonaea viscosa plant by using well and disc diffusion assay. The results indicated that well diffusion assay had best results than disc diffusion assay, the highest inhibition zone was 22 mm for well diffusion and 15 mm for disc diffusion assay, the results observed that ethanol extract had best antibacterial effect than aqueous extract which the percentage of bacterial isolates affected with ethanol extract was 71.19% comparing with aqueous extract 28.81% by using disc diffusion assay, while the percentage of bacterial isolates affected with ethanol extract was 88.13% comparing with aqueous extract 52.54% by using will diffusion assay.

Keywords: Salmonella spp, Dodonaea viscosa, antimicrobial and salmonellosis

Procedia PDF Downloads 444
1544 Evaluation of Heavy Metal Contamination and Assessment of the Suitability of Water for Irrigation: A Case Study of the Sand River, Limpopo Province, South Africa

Authors: Ngonidzashe Moyo, Mmaditshaba Rapatsa

Abstract:

The primary objective of this study was to determine heavy metal contamination in the water, sediment, grass and fish in Sand River, South Africa. This river passes through an urban area and sewage effluent is discharged into it. Water from the Sand river is subsequently used for irrigation downstream of the sewage treatment works. The suitability of this water and the surrounding boreholes for irrigation was determined. This study was undertaken between January, 2014 and January, 2015. Monthly samples were taken from four sites. Sites 1 was upstream of the Polokwane Wastewater Treatment Plant, sites 2, 3 and 4 were downstream. Ten boreholes in the vicinity of the Sand River were randomly selected and the water was tested for heavy metal contamination. The concentration of heavy metals in Sand River water followed the order Mn>Fe>Pb>Cu≥Zn≥Cd. Manganese concentration averaged 0.34 mg/L. Heavy metal concentration in the sediment, grass and fish followed the order Fe>Mn>Zn>Cu>Pb>Cd. The bioaccumulation factor from grass to fish was highest in manganese (19.25), followed by zinc (16.39) and iron (14.14). Soil permeability index (PI) and sodium adsorption ratio (SAR) were used to determine the suitability of Sand River and borehole water for irrigation. The PI index for Sand River water was 75.1% and this indicates that Sand River water is suitable for irrigation of crops. The PI index for the borehole water ranged from 65.8-72.8% and again this indicates suitability of borehole water for crop irrigation. The sodium adsorption ratio also indicated that both Sand River and borehole water were suitable for irrigation. A risk assessment study is recommended to determine the suitability of the fish for human consumption.

Keywords: bioaccumulation, bioavailability, heavy metals, sodium adsorption ratio

Procedia PDF Downloads 191
1543 Modeling Geogenic Groundwater Contamination Risk with the Groundwater Assessment Platform (GAP)

Authors: Joel Podgorski, Manouchehr Amini, Annette Johnson, Michael Berg

Abstract:

One-third of the world’s population relies on groundwater for its drinking water. Natural geogenic arsenic and fluoride contaminate ~10% of wells. Prolonged exposure to high levels of arsenic can result in various internal cancers, while high levels of fluoride are responsible for the development of dental and crippling skeletal fluorosis. In poor urban and rural settings, the provision of drinking water free of geogenic contamination can be a major challenge. In order to efficiently apply limited resources in the testing of wells, water resource managers need to know where geogenically contaminated groundwater is likely to occur. The Groundwater Assessment Platform (GAP) fulfills this need by providing state-of-the-art global arsenic and fluoride contamination hazard maps as well as enabling users to create their own groundwater quality models. The global risk models were produced by logistic regression of arsenic and fluoride measurements using predictor variables of various soil, geological and climate parameters. The maps display the probability of encountering concentrations of arsenic or fluoride exceeding the World Health Organization’s (WHO) stipulated concentration limits of 10 µg/L or 1.5 mg/L, respectively. In addition to a reconsideration of the relevant geochemical settings, these second-generation maps represent a great improvement over the previous risk maps due to a significant increase in data quantity and resolution. For example, there is a 10-fold increase in the number of measured data points, and the resolution of predictor variables is generally 60 times greater. These same predictor variable datasets are available on the GAP platform for visualization as well as for use with a modeling tool. The latter requires that users upload their own concentration measurements and select the predictor variables that they wish to incorporate in their models. In addition, users can upload additional predictor variable datasets either as features or coverages. Such models can represent an improvement over the global models already supplied, since (a) users may be able to use their own, more detailed datasets of measured concentrations and (b) the various processes leading to arsenic and fluoride groundwater contamination can be isolated more effectively on a smaller scale, thereby resulting in a more accurate model. All maps, including user-created risk models, can be downloaded as PDFs. There is also the option to share data in a secure environment as well as the possibility to collaborate in a secure environment through the creation of communities. In summary, GAP provides users with the means to reliably and efficiently produce models specific to their region of interest by making available the latest datasets of predictor variables along with the necessary modeling infrastructure.

Keywords: arsenic, fluoride, groundwater contamination, logistic regression

Procedia PDF Downloads 317
1542 Numerical Study on the Flow around a Steadily Rotating Spring: Understanding the Propulsion of a Bacterial Flagellum

Authors: Won Yeol Choi, Sangmo Kang

Abstract:

The propulsion of a bacterial flagellum in a viscous fluid has attracted many interests in the field of biological hydrodynamics, but remains yet fully understood and thus still a challenging problem. In this study, therefore, we have numerically investigated the flow around a steadily rotating micro-sized spring to further understand such bacterial flagellum propulsion. Note that a bacterium gains thrust (propulsive force) by rotating the flagellum connected to the body through a bio motor to move forward. For the investigation, we convert the spring model from the micro scale to the macro scale using a similitude law (scale law) and perform simulations on the converted macro-scale model using a commercial software package, CFX v13 (ANSYS). To scrutinize the propulsion characteristics of the flagellum through the simulations, we make parameter studies by changing some flow parameters, such as the pitch, helical radius and rotational speed of the spring and the Reynolds number (or fluid viscosity), expected to affect the thrust force experienced by the rotating spring. Results show that the propulsion characteristics depend strongly on the parameters mentioned above. It is observed that the forward thrust increases in a linear fashion with either of the rotational speed or the fluid viscosity. In addition, the thrust is directly proportional to square of the helical radius and but the thrust force is increased and then decreased based on the peak value to the pitch. Finally, we also present the appropriate flow and pressure fields visualized to support the observations.

Keywords: fluid viscosity, hydrodynamics, similitude, propulsive force

Procedia PDF Downloads 325
1541 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

Procedia PDF Downloads 157
1540 Bacterial Diversity in Human Intestinal Microbiota and Correlations with Nutritional Behavior, Physiology, Xenobiotics Intake and Antimicrobial Resistance in Obese, Overweight and Eutrophic Individuals

Authors: Thais O. de Paula, Marjorie R. A. Sarmiento, Francis M. Borges, Alessandra B. Ferreira-Machado, Juliana A. Resende, Dioneia E. Cesar, Vania L. Silva, Claudio G. Diniz

Abstract:

Obesity is currently a worldwide public health threat, being considered a pandemic multifactorial disease related to the human gut microbiota (GM). Add to that GM is considered an important reservoir of antimicrobial resistance genes (ARG) and little is known on GM and ARG in obesity, considering the altered physiology and xenobiotics intake. As regional and social behavior may play important roles in GM modulation, and most of the studies are based on small sample size and various methodological approaches resulting in difficulties for data comparisons, this study was focused on the investigation of GM bacterial diversity in obese (OB), overweight (OW) and eutrophic individuals (ET) considering their nutritional, clinical and social characteristics; and comparative screening of AGR related to their physiology and xenobiotics intake. Microbial community was accessed by FISH considering phyla as a taxonomic level, and PCR-DGGE followed by dendrograms evaluation (UPGMA method) from fecal metagenome of 72 volunteers classified according to their body mass index (BMI). Nutritional, clinical, social parameters and xenobiotics intake were recorded for correlation analysis. The fecal metagenome was also used as template for PCR targeting 59 different ARG. Overall, 62% of OB were hypertensive, and 12% or 4% were, regarding the OW and ET individuals. Most of the OB were rated as low income (80%). Lower relative bacterial densities were observed in the OB compared to ET for almost all studied taxa (p < 0.05) with Firmicutes/Bacteroidetes ratio increased in the OB group. OW individuals showed a bacterial density representative of GM more likely to the OB. All the participants were clustered in 3 different groups based on the PCR-DGGE fingerprint patterns (C1, C2, C3), being OB mostly grouped in C1 (83.3%) and ET mostly grouped in C3 (50%). The cluster C2 showed to be transitional. Among 27 ARG detected, a cluster of 17 was observed in all groups suggesting a common core. In general, ARG were observed mostly within OB individuals followed by OW and ET. The ratio between ARG and bacterial groups may suggest that AGR were more related to enterobacteria. Positive correlations were observed between ARG and BMI, calories and xenobiotics intake (especially use of sweeteners). As with nutritional and clinical characteristics, our data may suggest that GM of OW individuals behave in a heterogeneous pattern, occasionally more likely to the OB or to the ET. Regardless the regional and social behaviors of our population, the methodological approaches in this study were complementary and confirmatory. The imbalance of GM over the health-disease interface in obesity is a matter of fact, but its influence in host's physiology is still to be clearly elucidated to help understanding the multifactorial etiology of obesity. Although the results are in agreement with observations that GM is altered in obesity, the altered physiology in OB individuals seems to be also associated to the increased xenobiotics intake and may interfere with GM towards antimicrobial resistance, as observed by the fecal metagenome and ARG screening. Support: FAPEMIG, CNPQ, CAPES, PPGCBIO/UFJF.

Keywords: antimicrobial resistance, bacterial diversity, gut microbiota, obesity

Procedia PDF Downloads 135