Search results for: bacteria isolation

Authors: Nivethika Ajeethan, Lord Abbey, Svetlana Yurge

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Riboflavin is a water-soluble vitamin and the direct precursor of the flavin cofactors flavin mononucleotide and flavin adenine dinucleotide. Flavins (FLs) are bioactive molecules that have a beneficial effect on plant growth and development. Sinorhizobium meliloti strain 1021 is an α-proteobacterium that forms agronomically important N₂-fixing symbiosis with Medicago plants and secretes a considerable amount of FLs (FL⁺ strain). This strain was also implicated in plant growth promotion in its association with non-legume host plants. However, the mechanism of this plant growth promotion is not well understood. In this study, we evaluated the growth and development of tomato plants inoculated with S. meliloti 1021 and its mutant (FL⁻ strain) with limited ability to secrete FLs. Our preliminary experiments indicated that inoculation with FL⁺ strain significantly increased seedlings' root and shoot length and surface area compared to those of plants inoculated with FL⁻ strain. For example, the root lengths of 9-day old seedlings inoculated with FL⁺ strain were 35% longer than seedlings inoculated with the mutant. Proteomic approaches combined with the analysis of plant physiological responses such as growth and photosynthetic rate, stomatal conductance, transpiration rate, and chlorophyll content will be used to evaluate the host-plant response to bacteria-derived FLs.

Keywords: flavin, plant growth promotion, riboflavin, Sinorhizobium meliloti

Procedia PDF Downloads 154

Authors: D. Malouch, M. Berchel, C. Dreanno, S. Stachowski-Haberkorn, P-A. Jaffres

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Biofilm is a predominant lifestyle chosen by bacteria. Whether it is developed on an immerged surface or a mobile biofilm known as flocs, the bacteria within this form of life show properties different from its planktonic ones. Within the biofilm, the self-formed matrix of Extracellular Polymeric Substances (EPS) offers hydration, resources capture, enhanced resistance to antimicrobial agents, and allows cell-communication. Biofouling is a complex natural phenomenon that involves biological, physical and chemical properties related to the environment, the submerged surface and the living organisms involved. Bio-colonization of artificial structures can cause various economic and environmental impacts. The increase in costs associated with the over-consumption of fuel from biocolonized vessels has been widely studied. Measurement drifts from submerged sensors, as well as obstructions in heat exchangers, and deterioration of offshore structures are major difficulties that industries are dealing with. Therefore, surfaces that inhibit biocolonization are required in different areas (water treatment, marine paints, etc.) and many efforts have been devoted to produce efficient and eco-compatible antifouling agents. The different steps of surface fouling are widely described in literature. Studying the biofilm and its stages provides a better understanding of how to elaborate more efficient antifouling strategies. Several approaches are currently applied, such as the use of biocide anti-fouling paint6 (mainly with copper derivatives) and super-hydrophobic coatings. While these two processes are proving to be the most effective, they are not entirely satisfactory, especially in a context of a changing legislation. Nowadays, the challenge is to prevent biofouling with non-biocide compounds, offering a cost effective solution, but with no toxic effects on marine organisms. Since the micro-fouling phase plays an important role in the regulation of the following steps of biofilm formation7, it is desired to reduce or delate biofouling of a given surface by inhibiting the micro fouling at its early stages. In our recent works, we reported that some amphiphilic compounds exhibited bacteriostatic or bactericidal properties at a concentration that did not affect eukaryotic cells. These remarkable properties invited us to assess this type of bio-inspired phospholipids9 to prevent the colonization of surfaces by marine bacteria. Of note, other studies reported that amphiphilic compounds interacted with bacteria leading to a reduction of their development. An amphiphilic compound is a molecule consisting of a hydrophobic domain and a polar head (ionic or non-ionic). These compounds appear to have interesting antifouling properties: some ionic compounds have shown antimicrobial activity, and zwitterions can reduce nonspecific adsorption of proteins. Herein, we investigate the potential of amphiphilic compounds as inhibitors of bacterial growth and marine biofilm formation. The aim of this study is to compare the efficacy of four synthetic phospholipids that features a cationic charge (BSV36, KLN47) or a zwitterionic polar-head group (SL386, MB2871) to prevent microfouling with marine bacteria. We also study the toxicity of these compounds in order to identify the most promising compound that must feature high anti-adhesive properties and a low cytotoxicity on two links representative of coastal marine food webs: phytoplankton and oyster larvae.

Keywords: amphiphilic phospholipids, bacterial biofilm, marine microfouling, non-toxic antifouling

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Authors: Walaa Al-Maghraby

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Endophytes are organisms that colonize internal plant tissues without causing apparent harm to their host. Almost all groups of microorganisms have been found in endophytic association with plants may be fungi. They stimulate the production of secondary metabolites with a diverse range of biological activities. Leptadenia pyrotechnica is a more or less leafless, erect shrub with straight stems which is highly distributed in Saudi Arabia. Four endophytes fungi were isolated from Leptadenia pyrotechnica and identified using 18S ribosomal RNA sequences, which revealed four fungi genuses, namely Aspergillus terreus; Aspergillus welwitschiae; Aspergillus fumigatus and Aspergillus flavus. In this present study, four endophytic fungi from Leptadenia pyrotechnica were used for obtaining crude aqueous and ethyl acetate extracts for antimicrobial screening against 6 human pathogens, the antibacterial tests presented satisfactory results, where the pathogenic bacteria were inhibited by the four extracts tested, except for Escherichia coli that was inhibited by all extracts except ethyl acetate extract of Aspergillus terreus. Analysis of variance showed that the extract produced by endophyte Leptadenia pyrotechnica was the most effective against all bacteria, either gram-negative or positive. However, the extract was not efficient against pathogenic fungi. Therefore, this study indicates that endophytes from medicinal plant Leptadenia pyrotechnica could be potential sources of antibacterial substances.

Keywords: antimicrobial activity, Aspergillus sp, endophytes, Leptadenia pyrotechnica

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Authors: Danyi Wang, Andrew Schriefer, Dennis O'Rourke, Brajendra Kumar, Yang Liu, Fei Zhong, Juergen Scheuenpflug, Zheng Feng

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Purpose: It has been recognized that the microbiome plays critical roles in disease pathogenesis, including cancer, autoimmune disease, and multiple sclerosis. To develop a clinical-grade assay for exploring microbiome-derived clinical biomarkers across disease areas, a two-phase approach is implemented. 1) Identification of the optimal sample preparation reagents using pre-mixed bacteria and healthy donor stool samples coupled with proprietary Sigma-Aldrich® bioinformatics solution. 2) Exploratory analysis of patient samples for enabling precision medicine. Study Procedure: In phase 1 study, we first compared the 16S sequencing results of two ATCC® microbiome standards (MSA 2002 and MSA 2003) across five different extraction kits (Kit A, B, C, D & E). Both microbiome standards samples were extracted in triplicate across all extraction kits. Following isolation, DNA quantity was determined by Qubit assay. DNA quality was assessed to determine purity and to confirm extracted DNA is of high molecular weight. Bacterial 16S ribosomal ribonucleic acid (rRNA) amplicons were generated via amplification of the V3/V4 hypervariable region of the 16S rRNA. Sequencing was performed using a 2x300 bp paired-end configuration on the Illumina MiSeq. Fastq files were analyzed using the Sigma-Aldrich® Microbiome Platform. The Microbiome Platform is a cloud-based service that offers best-in-class 16S-seq and WGS analysis pipelines and databases. The Platform and its methods have been extensively benchmarked using microbiome standards generated internally by MilliporeSigma and other external providers. Data Summary: The DNA yield using the extraction kit D and E is below the limit of detection (100 pg/µl) of Qubit assay as both extraction kits are intended for samples with low bacterial counts. The pre-mixed bacterial pellets at high concentrations with an input of 2 x106 cells for MSA-2002 and 1 x106 cells from MSA-2003 were not compatible with the kits. Among the remaining 3 extraction kits, kit A produced the greatest yield whereas kit B provided the least yield (Kit-A/MSA-2002: 174.25 ± 34.98; Kit-A/MSA-2003: 179.89 ± 30.18; Kit-B/MSA-2002: 27.86 ± 9.35; Kit-B/MSA-2003: 23.14 ± 6.39; Kit-C/MSA-2002: 55.19 ± 10.18; Kit-C/MSA-2003: 35.80 ± 11.41 (Mean ± SD)). Also, kit A produced the greatest yield, whereas kit B provided the least yield. The PCoA 3D visualization of the Weighted Unifrac beta diversity shows that kits A and C cluster closely together while kit B appears as an outlier. The kit A sequencing samples cluster more closely together than both the other kits. The taxonomic profiles of kit B have lower recall when compared to the known mixture profiles indicating that kit B was inefficient at detecting some of the bacteria. Conclusion: Our data demonstrated that the DNA extraction method impacts DNA concentration, purity, and microbial communities detected by next-generation sequencing analysis. Further microbiome analysis performance comparison of using healthy stool samples is underway; also, colorectal cancer patients' samples will be acquired for further explore the clinical utilities. Collectively, our comprehensive qualification approach, including the evaluation of optimal DNA extraction conditions, the inclusion of positive controls, and the implementation of a robust qualified bioinformatics pipeline, assures accurate characterization of the microbiota in a complex matrix for deciphering the deep biology and enabling precision medicine.

Keywords: 16S rRNA sequencing, analytical validation, bioinformatics pipeline, metagenomics

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Authors: Azza A. Ali, Doaa M. Abd El-Latif, Amany M. Gad, Yasser M. A. Elnahas, Karema Abu-Elfotuh

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Background: Exposure to Aluminium (Al) has been increased recently. It is found in food products, food additives, drinking water, cosmetics and medicines. Chronic consumption of Al causes oxidative stress and has been implicated in several chronic disorders. Liver is considered as the major site for detoxification while kidney is involved in the elimination of toxic substances and is a target organ of metal toxicity. Social isolation (SI) or protein malnutrition (PM) also causes oxidative stress and has negative impact on Al-induced nephrotoxicity as well as hepatotoxicity. Coenzyme Q10 (CoQ10) is a powerful intracellular antioxidant with mitochondrial membrane stabilizing ability while wheat grass is a natural product with antioxidant, anti-inflammatory and different protective activities, cocoa is also potent antioxidants and can protect against many diseases. They provide different degrees of protection from the impact of oxidative stress. Objective: To study the impact of social isolation together with Protein malnutrition on nephro- and hepato-toxicity induced by chronic Al exposure in rats as well as to investigate the postulated protection using a combination of Co Q10, wheat grass and cocoa. Methods: Eight groups of rats were used; four served as protected groups and four as un-protected. Each of them received daily for five weeks AlCl3 (70 mg/kg, IP) for Al-toxicity model groups except one group served as control. Al-toxicity model groups were divided to Al-toxicity alone, SI- associated PM (10% casein diet) and Al- associated SI&PM groups. Protection was induced by oral co-administration of CoQ10 (200mg/kg), wheat grass (100mg/kg) and cocoa powder (24mg/kg) combination together with Al. Biochemical changes in total bilirubin, lipids, cholesterol, triglycerides, glucose, proteins, creatinine and urea as well as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate deshydrogenase (LDH) were measured in serum of all groups. Specimens of kidney and liver were used for assessment of oxidative parameters (MDA, SOD, TAC, NO), inflammatory mediators (TNF-α, IL-6β, nuclear factor kappa B (NF-κB), Caspase-3) and DNA fragmentation in addition to evaluation of histopathological changes. Results: SI together with PM severely enhanced nephro- and hepato-toxicity induced by chronic Al exposure. Co Q10, wheat grass and cocoa combination showed clear protection against hazards of Al exposure either alone or when associated with SI&PM. Their protection were indicated by the significant decrease in Al-induced elevations in total bilirubin, lipids, cholesterol, triglycerides, glucose, creatinine and urea levels as well as ALT, AST, ALP, LDH. Liver and kidney of the treated groups also showed significant decrease in MDA, NO, TNF-α, IL-6β, NF-κB, caspase-3 and DNA fragmentation, together with significant increase in total proteins, SOD and TAC. Biochemical results were confirmed by the histopathological examinations. Conclusion: SI together with PM represents a risk factor in enhancing nephro- and hepato-toxicity induced by Al in rats. CoQ10, wheat grass and cocoa combination provide clear protection against nephro- and hepatotoxicity as well as the consequent degenerations induced by chronic Al-exposure even when associated with the risk of SI together with PM.

Keywords: aluminum, nephrotoxicity, hepatotoxicity, isolation and protein malnutrition, coenzyme Q10, wheatgrass, cocoa, nutrients combinations

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Authors: M. Doudi, M. H. Pazandeh, L. Rahimzadeh Torabi

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Bedsores are pressure ulcers that occur on the skin or tissue due to being immobile and lying in bed for extended periods. Bedsores have the potential to progress into open ulcers, increasing the possibility of variety of bacterial infection. Acinetobacter baumannii, a pathogen of considerable clinical importance, exhibited a significant correlation with Bedsores (pressure ulcers) infections, thereby manifesting a wide spectrum of antibiotic resistance. The emergence of drug resistance has led researchers to focus on alternative methods, particularly phage therapy, for tackling bacterial infections. Phage therapy has emerged as a novel therapeutic approach to regulate the activity of these agents. The management of bacterial infections greatly benefits from the clinical utilization of bacteriophages as a valuable antimicrobial intervention. The primary objective of this investigation consisted of isolating and discerning potent bacteriophage capable of targeting multi drug-resistant (MDR) and extensively drug-resistant (XDR) bacteria obtained from pressure ulcers. In present study, analyzed and isolated A. baumannii strains obtained from a cohort of patients suffering from pressure ulcers at Taleghani Hospital in Ahvaz, Iran. An approach that included biochemical and molecular identification techniques was used to determine the taxonomic classification of bacterial isolates at the genus and species levels. The molecular identification process was facilitated by using the 16S rRNA gene in combination with universal primers 27 F, and 1492 R. Bacteriophage was obtained through the isolation process conducted on treatment plant sewage located in Isfahan, Iran. The main goal of this study was to evaluate different characteristics of phage, such as their appearance, range of hosts they can infect, how quickly they can enter a host, their stability at varying temperatures and pH levels, their effectiveness in killing bacteria, the growth pattern of a single phage stage, mapping of enzymatic digestion, and identification of proteomics patterns. The findings demonstrated that an examination was conducted on a sample of 50 specimens, wherein 15 instances of A. baumannii were identified. These microorganisms are the predominant Gram-negative agents known to cause wound infections in individuals suffering from bedsores. The study's findings indicated a high prevalence of antibiotic resistance in the strains isolated from pressure ulcers, excluding the clinical strains that exhibited responsiveness to colistin.According to the findings obtained from assessments of host range and morphological characteristics of bacteriophage VbɸAB-1, it can be concluded that this phage possesses specificity towards A. Baumannii BAH_Glau1001 was classified as a member of the Plasmaviridae family. The bacteriophage mentioned earlier showed the strongest antibacterial effect at a temperature of 18 °C and a pH of 6.5. Through the utilization of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis on protein fragments, it was established that the bacteriophage VbɸAB-1 exhibited a size range between 50 and 75 kilodaltons (KDa). The numerous research findings on the effectiveness of phages and the safety studies conducted suggest that the phages studied in this research can be considered as a practical solution and recommended approach for controlling and treating stubborn pathogens in burn wounds among hospitalized patients.

Keywords: acinetobacter baumannii, extremely drug- resistant, phage therapy, surgery wound

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Authors: Claudia L. Bianchi, Giuseppina Cerrato, Federico Galli, Federica Minozzi, Valentino Capucci

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At the beginning of the industrial productions, porcelain gres tiles were considered as just a technical material, aesthetically not very beautiful. Today thanks to new industrial production methods, both properties, and beauty of these materials completely fit the market requests. In particular, the possibility to prepare slabs of large sizes is the new frontier of building materials. Beside these noteworthy architectural features, new surface properties have been introduced in the last generation of these materials. In particular, deposition of TiO₂ transforms the traditional ceramic into a photocatalytic eco-active material able to reduce polluting molecules present in air and water, to eliminate bacteria and to reduce the surface dirt thanks to the self-cleaning property. The problem of photocatalytic materials resides in the fact that it is necessary a UV light source to activate the oxidation processes on the surface of the material, processes that are turned off inexorably when the material is illuminated by LED lights and, even more so, when we are in darkness. First, it was necessary a thorough study change the existing plants to deposit the photocatalyst very evenly and this has been done thanks to the advent of digital printing and the development of an ink custom-made that stabilizes the powdered TiO₂ in its formulation. In addition, the commercial TiO₂, which is used for the traditional photocatalytic coating, has been doped with metals in order to activate it even in the visible region and thus in the presence of sunlight or LED. Thanks to this active coating, ceramic slabs are able to purify air eliminating odors and VOCs, and also can be cleaned with very soft detergents due to the self-cleaning properties given by the TiO₂ present at the ceramic surface. Moreover, the presence of dopant metals (patent WO2016157155) also allows the material to work as well as antibacterial in the dark, by eliminating one of the negative features of photocatalytic building materials that have so far limited its use on a large scale. Considering that we are constantly in contact with bacteria, some of which are dangerous for health. Active tiles are 99,99% efficient on all bacteria, from the most common such as Escherichia coli to the most dangerous such as Staphilococcus aureus Methicillin-resistant (MRSA). DIGITALIFE project LIFE13 ENV/IT/000140 – award for best project of October 2017.

Keywords: Ag-doped microsized TiO₂, eco-active ceramic, photocatalysis, digital coating

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Authors: Guna Raj Dhungana, Roshan Nepal, Apshara Parajuli, , Archana Maharjan, Shyam K. Mishra, Pramod Aryal, Rajani Malla

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Introduction: Klebsiella pneumoniae is a well-known opportunistic human pathogen, primarily causing healthcare-associated infections. The global emergence of carbapenemase-producing K. pneumoniaeis a major public health burden, which is often extensively multidrug resistant.Thus, because of the difficulty to treat these ‘superbug’ and menace and some term as ‘apocalypse’ of post antibiotics era, an alternative approach to controlling this pathogen is prudent and one of the approaches is phage mediated control and/or treatment. Objective: In this study, we aimed to isolate novel bacteriophage against carbapenemase-producing K. pneumoniaeand characterize for potential use inphage therapy. Material and Methods: Twenty lytic phages were isolated from river water using double layer agar assay and purified. Biological features, physiochemical characters, burst size, host specificity and activity spectrum of phages were determined. One most potent phage: Phage TU_Kle10O was selected and characterized by electron microscopy. Whole genome sequences of the phage were analyzed for presence/absence of virulent factors, and other lysin genes. Results: Novel phage TU_Kle10O showed multiple host range within own genus and did not induce any BIM up to 5th generation of host’s life cycle. Electron microscopy confirmed that the phage was tailed and belonged to Caudovirales family. Next generation sequencing revealed its genome to be 166.2 Kb. bioinformatical analysis further confirmed that the phage genome ‘did not’ contain any ‘bacterial genes’ within phage genome, which ruled out the concern for transfer of virulent genes. Specific 'lysin’ enzyme was identified phages which could be used as 'antibiotics'. Conclusion: Extensively multidrug resistant bacteria like carbapenemase-producing K. pneumoniaecould be treated efficiently by phages.Absence of ‘virulent’ genes of bacterial origin and presence of lysin proteins within phage genome makes phages an excellent candidate for therapeutics.

Keywords: bacteriophage, Klebsiella pneumoniae, MDR, phage therapy, carbapenemase,

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Authors: Haftay Abraha Tadesse, Dawit Gebreegziabiher Hagos, Atsebaha Gebrekidan Kahsay, Mahumd Abdulkader

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Background: Salmonella species and Escherichia coli (E. coli) are important foodborne pathogens affecting humans and animals. They are among the most important causes of infection that are associated with the consumption of contaminated food. This study was aimed to determine the prevalence, antimicrobial susceptibility patterns and associated risk factors for Salmonella species and E. coli in raw meat from butchery houses of Mekelle, Northern Ethiopia. Method: A cross-sectional study was conducted from January to December 2019. Socio-demographic data and risk factors were collected using a predesigned questionnaire. Meat samples were collected aseptically from the butchery houses and transported using icebox to Mekelle University, College of Veterinary Sciences for the isolation and identification of Salmonella species and E. coli. Antimicrobial susceptibility patterns were determined using Kirby disc diffusion method. Data obtained were cleaned and entered into Statistical Package for the Social Sciences version 22 and logistic regression models with odds ratio were calculated. P-value < 0.05 was considered as statistically significant. Results: A total of 153 out of 384 (39.8%) of the meat specimens were found to be contaminated. The contamination of Salmonella species and E. coli were 15.6% (n=60) and 20.8%) (n=80), respectively. Mixed contamination (Salmonella species and E. coli) was observed in 13 (3.4 %) of the analyzed. Poor washing hands regularly (AOR = 8.37; 95% CI: 2.75-25.50) and not using gloves during meat handling (AOR=11. 28; 95% CI:(4.69 27.10) were associated with overall bacterial contamination. About 100% of the tested isolates were sensitive to ciprofloxacin, gentamicin, Co trimoxazole , sulphamethoxazole, ceftriaxone, and trimethoprim and ciprofloxacin, gentamicin, and norfloxacine of E. coli and Salmonella species, respectively, while the resistance of amoxyclav_amoxicillin and erythromycin were both isolated bacteria species. The overall multidrug resistance pattern for Salmonella and E. coli were 51.4% (n=19) and 31.8% (14), respectively. Conclusion: Of the 153 (153/384) contaminated raw meat, 60 (15.6%) and 80 (20.8%) were contaminated by Salmonella species and E. coli, respectively. Poor handwashing practice and not using glove during meat handling showed a significant association with bacterial contamination. Multidrug-resistant showed in Salmonella species, and E. coli were 19 (51.4%) and 14 (31.8%), respectively.

Keywords: antimicrobial susceptibility test, butchery houses, E. coli, raw meat, salmonella species

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Authors: Ali Bahkali, Rayan Yousef Booq, Mohammad Khiyami

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Soil samples were collected from five different regions in the Kingdom of Saudi Arabia. Microbiological methods included dilution methods and pour plates to isolate and purify bacteria soil. The ability of isolates to develop biopolymer was investigated on petri dishes containing elements and substance concentrations stimulating developing biopolymer. Fluorescent stains, Nile red and Nile blue were used to stain the bacterial cells developing biopolymers. In addition, Sudan black was used to detect biopolymers in bacterial cells. The isolates which developed biopolymers were identified based on their gene sequence of 1 6sRNA and their ability to grow and synthesize PHAs on mineral medium supplemented with 1% dates molasses as the only carbon source under nitrogen limitation. During the study 293 bacterial isolates were isolated and detected. Through the initial survey on the petri dishes, 84 isolates showed the ability to develop biopolymers. These bacterial colonies developed a pink color due to accumulation of the biopolymers in the cells. Twenty-three isolates were able to grow on dates molasses, three strains of which showed the ability to accumulate biopolymers. These strains included Bacillus sp., Ralstonia sp. and Microbacterium sp. They were detected by Nile blue A stain with fluorescence microscopy (OLYMPUS IX 51). Among the isolated strains Ralstonia sp. was selected after its ability to grow on molasses dates in the presence of a limited nitrogen source was detected. The optimum conditions for formation of biopolymers by isolated strains were investigated. Conditions studied included, best incubation duration (2 days), temperature (30°C) and pH (7-8). The maximum PHB production was raised by 1% (v1v) when using concentrations of dates molasses 1, 2, 3, 4 and 5% in MSM. The best inoculated with 1% old inoculum (1= OD). The ideal extraction method of PHA and PHB proved to be 0.4% sodium hypochlorite solution, producing a quantity of polymer 98.79% of the cell's dry weight. The maximum PHB production was 1.79 g/L recorded by Ralstonia sp. after 48 h, while it was 1.40 g/L produced by R.eutropha ATCC 17697 after 48 h.

Keywords: bacteria forming polyhydroxyalkanoate, detection, molecular, Saudi Arabia

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Authors: Abdul Musaweer Habib, Habibul Hasan Mazumder, Saiful Islam, Sohel Sikder, Omar Faruk Sikder

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The plethora of genome sequence information of bacteria in recent times has ushered in many novel strategies for antibacterial drug discovery and facilitated medical science to take up the challenge of the increasing resistance of pathogenic bacteria to current antibiotics. In this study, we adopted subtractive genomics approach to analyze the whole genome sequence of the Fusobacterium nucleatum, a human oral pathogen having association with colorectal cancer. Our study divulged 1499 proteins of Fusobacterium nucleatum, which has no homolog in human genome. These proteins were subjected to screening further by using the Database of Essential Genes (DEG) that resulted in the identification of 32 vitally important proteins for the bacterium. Subsequent analysis of the identified pivotal proteins, using the KEGG Automated Annotation Server (KAAS) resulted in sorting 3 key enzymes of F. nucleatum that may be good candidates as potential drug targets, since they are unique for the bacterium and absent in humans. In addition, we have demonstrated the 3-D structure of these three proteins. Finally, determination of ligand binding sites of the key proteins as well as screening for functional inhibitors that best fitted with the ligands sites were conducted to discover effective novel therapeutic compounds against Fusobacterium nucleatum.

Keywords: colorectal cancer, drug target, Fusobacterium nucleatum, homology modeling, ligands

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Authors: Waode Nurmayani, Nikmatul Riswanda

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Broilers are chickens which are kept with the most efficient time and hoped get a good body weight. All things are done, for example with the improvement of feed and use antibiotics. Feed cost is the most cost to be spent. Nearly 80% of the cost is spent just for buy feed. Earthworm (Lumbricus rubellus) is a good choice to reduce the cost of feed protein source. The Earthworm has a high crude protein content of about 48.5%-61.9%, rich with proline amino acid about 15% of the 62 amino acids. Not only about protein, this earthworm also has a role in disease prevention. Prevention of disease in livestock usual with use feed supplement. Earthworm (Lumbricus rubellus) is one of the natural materials used as feed. In addition, several types of earthworms that have been known to contain active substances about antibacterial pathogens namely Lumbricus rubellus. The earthworm could be used as an antibiotic because it contain the antibody of Lumbricine active substance. So that, this animal feed from Lumbricus rubellus could improve the performance of broilers. Bioactive of anti-bacterial is called Lumbricine able to inhibit the growth of pathogenic bacteria in the intestinal wall so that the population of pathogenic bacteria is reduced. The method of write in this scientific writing is divided into 3 techniques, namely data completion, data analysis, and thinking pan from various literature about earthworm (Lumbricus rubellus) as broiler feed. It is expected that innovation of feed material of earthworm (Lumbricus rubellus) could reduce the cost of protein feed and the use of chemical antibiotics.

Keywords: earthworm, broiler, protein, antibiotic

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Authors: Maria Luisa de la Torre, Javier Aroba, Jose Miguel Davila, Aguasanta M. Sarmiento

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Steel is one of the most widely used materials in polymetallic sulfide mining installations. One of the main problems suffered by these facilities is the economic losses due to the corrosion of this material, which is accelerated and aggravated by the contact with acid waters generated in these mines when sulfides come into contact with oxygen and water. This generation of acidic water, in turn, is accelerated by the presence of acidophilic bacteria. In order to gain a more detailed understanding of this corrosion process and the interaction between steel and acidic water, a laboratory experiment was carried out in which carbon steel plates were introduced into four different solutions for 27 days: distilled water (BK), which tried to assimilate the effect produced by rain on this material, an acid solution from a mine with a high Fe2+/Fe3+ (PO) content, another acid solution of water from another mine with a high Fe3+/Fe2+ (PH) content and, finally, one that reproduced the acid mine water with a high Fe2+/Fe3+ content but in which there were no bacteria (ST). Every 24 hours, physicochemical parameters were measured, and water samples were taken to carry out an analysis of the dissolved elements. The results of these measurements were processed using an explainable AI model based on fuzzy logic. It could be seen that, in all cases, there was an increase in pH, as well as in the concentrations of Fe and, in particular, Fe(II), as a consequence of the oxidation of the steel plates. Proportionally, the increase in Fe concentration was higher in PO and ST than in PH because Fe precipitates were produced in the latter. The rise of Fe(II) was proportionally much higher in PH, especially in the first hours of exposure, because it started from a lower initial concentration of this ion. Although to a lesser extent than in PH, the greater increase in Fe(II) also occurred faster in PO than in ST, a consequence of the action of the catalytic bacteria. On the other hand, Cu concentrations decreased throughout the experiment (with the exception of distilled water, which initially had no Cu, as a result of an electrochemical process that generates a precipitation of Cu together with Fe hydroxides. This decrease is lower in PH because the high total acidity keeps it in solution for a longer time. With the application of an artificial intelligence tool, it has been possible to evaluate the effects of steel corrosion in mining environments, corroborating and extending what was obtained by means of classical statistics.

Keywords: acid mine drainage, artificial intelligence, carbon steel, corrosion, fuzzy logic

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Authors: Dagim Ali Hussen, Adnan A. Bekhit, Ariaya Hymete

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In this study, several pyridine derivatives were synthesized and evaluated for their in vitro antimicrobial activity against gram-positive bacteria (S. aureus and B. Cereus), gram-negative bacteria (P. aeruginosa and E. coli) and fungus (C. albican and A niger). The intermediate chalcone derivative 2a,b was synthesized by condensation of pyrazole aldehydes 1a,b with acetophenone in alcoholic KOH. Cyclization of 2a,b with ethyl cyanoacetate ad ammonium acetate resulted in pyridine carbonitrile derivatives 3a,b. Furthermore, condensation of pyridine-4-carboxaldeyhe with different amino-derivatives gave rise to pyridine derivatives 5a,b, 6a,b. The oxadiazole derivative 7a was prepared by cyclization of 6a with acetic anhydride. Characterization of the synthesized compound was performed using IR, 1H NMR, 13C NMR spectra and elemental microanalyses. The antimicrobial results revealed that compounds 5a, 6b and 7a exhibited half fold antibacterial activity compared to ampicillin, against B. cereus. On the other hand, compound 3b showed an equivalent activity compared to miconazole against candida albican (CANDAL 03) and to clotrimazole against the clinical isolate candida albican 6647. Moreover, this compound 3b was further tested for its acute toxicity profile. The results showed that oral LD50 is more that 300 mg/kg and parentral LD50 is more than 100 mg/kg. Compound 3b is a good candidate for antifungal agent with good toxicity profile, and deserves more chemical derivatization and clinical study.

Keywords: antifungal, antimicrobial, Candida albican, pyridine

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Authors: Mohammad Hossein Pazandeh, Monir Doudi, Sona Rostampour Yasouri

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—Results The prudent administration of antibiotics aims to avoid the side effects and the microbes' resistance to antibiotics. An approach developing methods of local administration of antibiotics is especially required for localized infections caused by bacterial colonization of medical devices or implant materials. Among the wide variety of materials used as drug delivery systems, bioactive glasses (BG) have large utilization in regenerative medicine . firstly, the production of bioactive glass/nickel oxide/tin dioxide nanocomposite using sol-gel method, and then, the controlled release of imipenem from the double metal oxide/bioactive glass nanocomposite, and finally, the investigation of the antibacterial property of the nanocomposite. against a number of implant-related infectious agents. In this study, BG/SnO2 and BG/NiO single systema with different metal oxide present and BG/NiO/SnO2 nanocomposites were synthesized by sol-gel as drug carriers for tetracycline and imepinem. These two antibiotics were widely used for osteomyelitis because of its favorable penetration and bactericidal effect on all the probable osteomyelitis pathogens. The antibacterial activity of synthesized samples were evaluated against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa as bacteria model using disk diffusion method. The BG modification using metal oxides results to antibacterial property of samples containing metal oxide with highest efficiency for nancomposite. bioactivity of all samples was assessed by determining the surface morphology, structural and composition changes using scanning electron microscopy (SEM), FTIR and X-ray diffraction (XRD) spectroscopy, respectively, after soaking in simulated body fluid (SBF) for 28 days. The hydroxyapatite formation was clearly observed as a bioactivity measurement. Then, BG nanocomposite sample was loaded using two antibiotics, separately and their release profiles were studied. The BG nancomposite sample was shown the slow and continuous drug releasing for a period of 72 hours which is desirable for a drug delivery system. The loaded antibiotic nanocomposite sample retaining antibacterial property and showing inactivation effect against bacteria under test. The modified bioactive glass forming hydroxyapatite with controlled release drug and effective against bacterial infections can be introduced as scaffolds for bone implants after clinical trials for biomedical applications . Considering the formation of biofilm by infectious bacteria after sticking on the surfaces of implants, medical devices, etc. Also, considering the complications of traditional methods, solving the problems caused by the above-mentioned microorganisms in technical and biomedical industries was one of the necessities of this research.

Keywords: antibacterial, bioglass, drug delivery system, sol- gel

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Authors: Amalia Dwi Berliyanti Amel

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Streptococcus mutants bacteria are bacteria that are believed to be the cause of the growth of dental plaque which can further adversely affect dental caries if left unchecked. Previous research has shown that Moringa leaf extract can slow down the growth rate of this bacterium. This study aims to make the best formulation of mouthwash with the active ingredient of Moringa leaf extract based on its antibacterial and organoleptic test results. Nine mouthwash variations were carried out with two factors and three levels, namely a comparison of the concentration of sorbitol (A) with three levels namely 15% (A1), 20% (A2), and 25% (A3), and peppermint added (B) with three levels, namely 0.2% (B1), 0.25% (B2), and 0.3% (B3). The test parameters performed as the determination of the best mouthwash are based on physicochemical properties which include pH and viscosity as well as organoleptic test results which include color, viscosity, aroma, taste, sensation in the mouth, and general appearance. The results showed that the bright zone as a test for the antibacterial activity of Streptococcus mutants began to be seen at a concentration of 5%. Moringa leaf mouthwash formulation has a pH value between 6 - 7, with a control of 6. Whereas the mucosa leaf mouthwash vascularity produced between 1.1 - 1.7 cP with a control of 1.1 cP. Moringa leaf mouthwash and control have the same total number of microbes, namely 0 colonies / mL. Based on organoleptic tests performed with 20 panelists, it was shown that the best mouthwash formulation was formulation A1B3 with sorbitol composition 15% and peppermint 0.3%.

Keywords: antibasteria, formula, moringa leaf, mouthwash

Procedia PDF Downloads 155

Authors: Fayad Ghawbar

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The significance of the Multiple Input Multiple Output (MIMO) system in the 5G wireless communication system is essential to enhance channel capacity and provide a high data rate resulting in a need for dual-polarization in vertical and horizontal. Furthermore, size reduction is critical in a MIMO system to deploy more antenna elements requiring a compact, low-profile design. A compact dual-band 4-MIMO antenna system has been presented in this paper with pattern and polarization diversity. The proposed single antenna structure has been designed using two antenna layers with a C shape in the front layer and a partial slot with a U-shaped cut in the ground to enhance isolation. The single antenna is printed on an FR4 dielectric substrate with an overall size of 18 mm×18 mm×1.6 mm. The 4-MIMO antenna elements were printed orthogonally on an FR4 substrate with a size dimension of 36 × 36 × 1.6 mm3 with zero edge-to-edge separation distance. The proposed compact 4-MIMO antenna elements resonate at 3.4-3.6 GHz and 4.8-5 GHz. The s-parameters measurement and simulation results agree, especially in the lower band with a slight frequency shift of the measurement results at the upper band due to fabrication imperfection. The proposed design shows isolation above -15 dB and -22 dB across the 4-MIMO elements. The MIMO diversity performance has been evaluated in terms of efficiency, ECC, DG, TARC, and CCL. The total and radiation efficiency were above 50 % across all parameters in both frequency bands. The ECC values were lower than 0.10, and the DG results were about 9.95 dB in all antenna elements. TARC results exhibited values lower than 0 dB with values lower than -25 dB in all MIMO elements at the dual-bands. Moreover, the channel capacity losses in the MIMO system were depicted using CCL with values lower than 0.4 Bits/s/Hz.

Keywords: compact antennas, MIMO antenna system, 5G communication, dual band, ECC, DG, TARC

Procedia PDF Downloads 143

Authors: M. Chukhryaeva, R. Skhalyakho, J. Kagazegeva, E. Pocheshkhova, L. Yepiskopossyan, O. Balanovsky, E. Balanovska

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The Middle East is crossroad of different populations at different times. The Kurds are of particular interest in this region. Historical sources suggested that the origin of the Kurds is associated with Medes. Therefore, it was especially interesting to compare gene pool of Kurds with other supposed descendants of Medes-Tats. Yezidis are ethno confessional group of Kurds. Yezidism as a confessional teaching was formed in the XI-XIII centuries in Iraq. Yezidism has caused reproductively isolation of Yezidis from neighboring populations for centuries. Also, isolation helps to retain Yezidian caste system. It is unknown how the history of Yezidis affected its genу pool because it has never been the object of researching. We have examined the Y-chromosome variation in Yezidis and Kurdish males to understand their gene pool. We collected DNA samples from 90 Yezidi males and 24 Kurdish males together with their pedigrees. We performed Y-STR analysis of 17 loci in the samples collected (Yfiler system from Applied Biosystems) and analysis of 42 Y-SNPs by real-time PCR. We compared our data with published data from other Kurdish groups and from European, Caucasian, and West Asian populations. We found that gene pool of Yezidis contains haplogroups common in the Middle East (J-M172(xM67,M12)- 24%, E-M35(xM78)- 9%) and in South Western Asia (R-M124- 8%) and variant with wide distribution area - R-M198(xM458- 9%). The gene pool of Kurdish has higher genetic diversity than Yezidis. Their dominants haplogroups are R-M198- 20,3 %, E-M35- 9%, J-M172- 9%. Multidimensional scaling also shows that the Kurds and Yezidis are part of the same frontier Asian cluster, which, in addition, included Armenians, Iranians, Turks, and Greeks. At the same time, the peoples of the Caucasus and Europe form isolated clusters that do not overlap with the Asian clusters. It is noteworthy that Kurds from our study gravitate towards Tats, which indicates that most likely these two populations are descendants of ancient Medes population. Multidimensional scaling also reveals similarity between gene pool of Yezidis, Kurds with Armenians and Iranians. The analysis of Yezidis pedigrees and their STR variability did not reveal a reliable connection between genetic diversity and caste system. This indicates that the Yezidis caste system is a social division and not a biological one. Thus, we showed that, despite many years of isolation, the gene pool of Yezidis retained a common layer with the gene pool of Kurds, these populations have common spectrum of haplogroups, but Yezidis have lower genetic diversity than Kurds. This study received primary support from the RSF grant No. 16-36-00122 to MC and grant No. 16-06-00364 to EP.

Keywords: gene pool, haplogroup, Kurds, SNP and STR markers, Yezidis

Procedia PDF Downloads 205

Authors: M. Doudi, M. H. Pazandeh, L. Rahimzadeh Torabi

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Bedsores are pressure ulcers that occur on the skin or tissue due to being immobile and lying in bed for extended periods. Bedsores have the potential to progress into open ulcers, increasing the possibility of a variety of bacterial infections. Acinetobacter baumannii, a pathogen of considerable clinical importance, exhibited a significant correlation with Bedsores (pressure ulcers) infections, thereby manifesting a wide spectrum of antibiotic resistance. The emergence of drug resistance has led researchers to focus on alternative methods, particularly phage therapy, for tackling bacterial infections. Phage therapy has emerged as a novel therapeutic approach to regulate the activity of these agents. The management of bacterial infections greatly benefits from the clinical utilization of bacteriophages as a valuable antimicrobial intervention. The primary objective of this investigation consisted of isolating and discerning potent bacteriophage capable of targeting multi-drug-resistant (MDR) and extensively drug-resistant (XDR) bacteria obtained from pressure ulcers. The present study analyzed and isolated A. baumannii strains obtained from a cohort of patients suffering from pressure ulcers at Taleghani Hospital in Ahvaz, Iran. An approach that included biochemical and molecular identification techniques was used to determine the taxonomic classification of bacterial isolates at the genus and species levels. The molecular identification process was facilitated by using the 16S rRNA gene in combination with universal primers 27 F and 1492 R. Bacteriophage was obtained through the isolation process conducted on treatment plant sewage located in Isfahan, Iran. The main goal of this study was to evaluate different characteristics of phage, such as their appearance, the range of hosts they can infect, how quickly they can enter a host, their stability at varying temperatures and pH levels, their effectiveness in killing bacteria, the growth pattern of a single phage stage, mapping of enzymatic digestion, and identification of proteomics patterns. The findings demonstrated that an examination was conducted on a sample of 50 specimens, wherein 15 instances of A. baumannii were identified. These microorganisms are the predominant Gram-negative agents known to cause wound infections in individuals suffering from bedsores. The study's findings indicated a high prevalence of antibiotic resistance in the strains isolated from pressure ulcers, excluding the clinical strains that exhibited responsiveness to colistin. According to the findings obtained from assessments of host range and morphological characteristics of bacteriophage VbɸAB-1, it can be concluded that this phage possesses specificity towards A. Baumannii BAH_Glau1001 was classified as a member of the Podoviridae family. The bacteriophage mentioned earlier showed the strongest antibacterial effect at a temperature of 18 °C and a pH of 6.5. Through the utilization of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis on protein fragments, it was established that the bacteriophage VbɸAB-1 exhibited a size range between 50 and 75 kilodaltons (KDa). The numerous research findings on the effectiveness of phages and the safety studies conducted suggest that the phages studied in this research can be considered as a practical solution and recommended approach for controlling and treating stubborn pathogens in burn wounds among hospitalized patients. The findings of our research indicated that isolated phages could be an effective antimicrobial and an appreciate candidate for prophylaxis against pressure ulcers.

Keywords: acinetobacter baumannii, extremely drug-resistant, phage therapy, surgery wound

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Authors: Ikram Kamal, Mohamed Blaghen

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Biosurfactants are amphiphilic biological compounds consisting of hydrophobic and hydrophilic domains produced extracellularly or as part of the cell membrane by a variety of yeast, bacteria and filamentous fungi. Biosurfactant applications in the environmental industries are promising due to their biodegradability, low toxicity, and effectiveness in enhancing biodegradation and solubilization of low solubility compounds. Currently, the main application is for enhancement of oil recovery and hydrocarbon bioremediation due to their biodegradability and low critical micelle concentration (CMC). The use of biosurfactants has also been proposed for various industrial applications, such as in food additives, cosmetics, detergent formulations and in combinations with enzymes for wastewater treatment. In this study, we have investigated the potential of bacterial strains: Mannheimia haemolytica, Burkholderia cepacia and Serratia ficaria were collected aseptically from the lagoon Marchika (water and soil) in Nador, Morocco; for the production of biosurfactants. This study also aimed to optimize the biosurfactant production process by changing the variables that influence the type and amount of biosurfactant produced by these microorganisms such as: carbon sources and also other physical and chemical parameters such as temperature and pH. Emulsification index, methylene blue test, and thin layer chromatography (TLC) revealed the ability of strains used in this study to produce compounds that could emulsify gasoline. In addition, a GC/MS was used to separate and identify different biosurfactants purified.

Keywords: biosurfactants, Mannheimia haemolytica, biodegradability, Burkholderia cepacia, Serratia ficaria

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Authors: Benguerine Zohra, Merzak Siham, Pr. Chelghoum

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Introduction: The widespread production and consumption of antibiotics have raised significant concerns due to various adverse effects and the development of bacterial resistance. This increasing resistance to currently available antibiotics necessitates the search for new antibacterial agents. One alternative strategy to combat antibiotic-resistant bacteria is the use of natural antimicrobial substances such as plant extracts. This study aims to provide an overview of the antibacterial effects of Salvia officinalis (sage), a plant native to the Middle East and Mediterranean regions. Materials and Methods: This review was conducted by searching studies in databases such as PubMed, Scopus, JSTOR, and SpringerLink. The search terms were “Salvia officinalis L.” and “antibacterial effects.” Only studies that met our inclusion criteria (in English, focusing on the antibacterial effects of Salvia officinalis L., and primarily dated from 2012 to 2023) were considered for further review. Results and Discussion: The initial search strategy identified approximately 78 references, of which only 13 articles were included in this review. The synthesis of these articles revealed that multiple data sources confirm the antimicrobial effects of S. officinalis. Its essential oil and alcoholic extract exhibit strong bactericidal and bacteriostatic effects against both Gram-positive and Gram-negative bacteria. Conclusion: The significant value of the extract, oil, and leaves of S. officinalis demands further studies on other useful and unknown properties of this multipurpose plant.

Keywords: salvia officinalis, literature review, antibacterial., botany

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Authors: Natapol Pumipuntu

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Staphylococcus argenteus is the emerging species of S. aureus complex. It was generally misidentified as S. aureus by standard techniques and their features. S. argenteus is possibly emerging in both humans and animals, as well as increasing worldwide distribution. The objective of this study was to differentiate and identify S. argenteus from S. aureus, which has been collected and isolated from milk samples of subclinical bovine mastitis cases in Maha Sarakham province, Northeastern of Thailand. Twenty-one isolates of S. aureus, which confirmed by conventional methods and immune-agglutination method were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and multilocus sequence typing (MLST). The result from MALDI-TOF MS and MLST showed 6 from 42 isolates were confirmed as S. argenteus, and 36 isolates were S. aureus, respectively. This study indicated that the identification and classification method by using MALDI-TOF MS and MLST could accurately differentiate the emerging species, S. argenteus, from S. aureus complex which usually misdiagnosed. In addition, the identification of S. argenteus seems to be very limited despite the fact that it may be the important causative pathogen in bovine mastitis as well as pathogenic bacteria in food and milk. Therefore, it is very necessary for both bovine medicine and veterinary public health to emphasize and recognize this bacterial pathogen as the emerging disease of Staphylococcal bacteria and need further study about S. argenteus infection.

Keywords: Staphylococcus argenteus, subclinical bovine mastitis, Staphylococcus aureus complex, mass spectrometry, MLST

Procedia PDF Downloads 151

Authors: Retno A. S. Lestari, Wahyudi B. Sediawan, Siti Syamsiah, Sarto

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Sulfur-oxidizing bacteria were isolated and then grown on snakefruits seeds forming biofilm. Their performance in sulfide removal were experimentally observed. Snakefruit seeds were then used as packing material in a cylindrical tube. Biological treatment of hydrogen sulfide from biogas was investigated using biofilm on packed bed of snakefruits seeds. Biogas containing 27,9512 ppm of hydrogen sulfide was flown through the bed. Then the hydrogen sulfide concentrations in the outlet at various times were analyzed. A set of simple kinetics model for the rate of the sulfide removal and the bacterial growth was proposed. The axial sulfide concentration gradient in the flowing liquid are assumed to be steady-state. Mean while the biofilm grows on the surface of the seeds and the oxidation takes place in the biofilm. Since the biofilm is very thin, the sulfide concentration in the biofilm is assumed to be uniform. The simultaneous ordinary differential equations obtained were then solved numerically using Runge-Kutta method. The acuracy of the model proposed was tested by comparing the calcultion results using the model with the experimental data obtained. It turned out that the model proposed can be applied to describe the removal of sulfide liquid using bio-filter in packed bed. The values of the parameters were also obtained by curve-fitting. The biofilter could remove 89,83 % of the inlet of hydrogen sulfide from biogas for 2.5 h, and optimum loading of 8.33 ml/h.

Keywords: Sulfur-oxidizing bacteria, snakefruits seeds, biofilm, packing material, biogas

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Authors: Ahmed Tawfik

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Oxidation of 1,4 dioxane produces metabolites by-products involving glycolaldehyde and acids that have geno- and cytotoxicity impact on microbial degradation. Thereby, the incorporation of algae with bacteria in the treatment system would eliminate and overcome the accumulation of metabolites that are utilized as a carbon source for the build-up of biomass. Therefore, the aim of the present study is to assess the potential of algae/bacteria-based membrane bioreactor (AB-MBR) for biodegradation of 1,4 dioxane-rich wastewater at a high imposed loading rate. Three identical reactors, i.e., AB-MBR1, AB-MBR2, and AB-MBR3, were operated in parallel at 1,4 dioxane loading rates of 641.7, 320.9, and 160.4 mg/L. d., and HRTs of 6.0, 12 and 24 h. respectively. The AB-MBR1 achieved 1,4 dioxane removal rate of 263.7 mg/L.d., where the residual value in the treated effluent amounted to 94.4±22.9 mg/L. Reducing the 1,4 dioxane loading rate (LR) to 320.9 mg/L.d in the AB-MBR2 maximized the removal rate efficiency of 265.9 mg/L.d., with a removal efficiency of 82.8±3.2%. The minimum value of 1,4 dioxane of 17.3±1.8 mg/L in the treated effluent of AB-MBR3 was obtained at an HRT of 24.0 h and loading rate of 160.4 mg/L.d. The mechanism of 1,4 dioxane degradation in AB-MBR was a combination of volatilization (8.03±0.6%), UV oxidation (14.1±0.9%), microbial biodegradation (49.1±3.9%) and absorption/uptake and assimilation by algae (28.8±2.%). Further, the Thioclava, Afipia, and Mycobacterium genera oxidized and produced the required enzymes for hydrolysis and cleavage of the dioxane ring into 2-hydroxy-1,4 dioxane. Moreover, the fungi, i.e., Basidiomycota and Cryptomycota, played a big role in the degradation of the 1,4 dioxane into 2-hydroxy-1,4 dioxane. Xanthobacter and Mesorhizobium were involved in the metabolism process by secreting alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH), and glycolate oxidase. Bacteria and fungi produced dehydrogenase (DH) for the transformation of 2-hydroxy-1,4 dioxane into 2-hydroxy-ethoxyacetaldehyde. The latter is converted into Ethylene glycol by Aldehyde hydrogenase (ALDH). Ethylene glycol is oxidized into acids using Alcohol hydrogenase (ADH). The Diatomea, Chlorophyta, and Streptophyta utilize the metabolites for biomass assimilation and produce the required oxygen for further oxidation of the dioxane and its metabolites by-products of bacteria and fungi. The major portion of metabolites (ethylene glycol, glycolic acid, and oxalic acid were removed due to uptake and absorption by algae (43±4.3%), followed by adsorption (18.4±0.9%). The volatilization and UV oxidation contribution for the degradation of metabolites were 8.7±0.7% and 12.3±0.8%, respectively. The capabilities of genera Defluviimonas, Thioclava, Luteolibacter, and Afipia. The genera of Defluviimonas, Thioclava, Luteolibacter, and Mycobacterium were grown under a high 1,4 dioxane LR of 641.7 mg/L.d. The Chlorophyta (4.1-43.6%), Streptophyta (2.5-21.7%), and Diatomea (0.8-1.4%) phyla were dominant for degradation of 1,4 dioxane. The results of this study strongly demonstrated that the bioremediation and bioaugmentation process can safely remove 1,4 dioxane from industrial wastewater while minimizing environmental concerns and reducing economic costs.

Keywords: wastewater, membrane bioreactor, bacterial community, algal community

Procedia PDF Downloads 43

Authors: Rajeev Manhas, M. A. Bhat, A. K. Taku, Dalip Singh, Deep Shikha, Gulzar Bader

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The study was aimed to understand the distribution of various serogroups of Pasteurella multocida in bovines, small ruminants, pig, rabbit, and poultry from Jammu, Jammu and Kashmir and to characterize the isolates with respect to LPS synthesizing genes, dermonecrotic toxin gene (toxA) gene and antibiotic resistance. For isolation, the nasopharyngeal swab procedure appeared to be better than nasal swab procedure, particularly in ovine and swine. Out of 200 samples from different animals, isolation of P. multocida could be achieved from pig and sheep (5 each) and from poultry and buffalo (2 each) samples only, which accounted for 14 isolates. Upon molecular serogrouping, 3 isolates from sheep and 2 isolates from poultry were found as serogroup A, 2 isolates from buffalo were confirmed as serogroup B and 5 isolates from pig were found to belong to serogroup D. However, 2 isolates from sheep could not be typed, hence, untypable. All the 14 isolates were subjected to mPCR genotyping. A total of 10 isolates, 5 each from pig and sheep, generated an amplicon specific to genotype L6 and L6 indicates Heddleston serovars 10, 11, 12 and 15. Similarly, 2 isolates from bovines generated an amplicon of genotype L2 which indicates Heddleston serovar 2/5. However, 2 isolates from poultry generated specific amplicon with L1 signifying Heddleston serovar 1, but these isolates also produced multiple bands with primer L5. Only, one isolate of capsular type A from sheep possessed the structural gene, toxA for dermonecrotoxin. There was variability in the antimicrobial susceptibility pattern in sheep isolates, but overall the rate of tetracycline resistance was relatively high (64.28%) in our strains while all the isolates were sensitive to streptomycin. Except for the swine isolates and one toxigenic sheep isolate, the P. multocida isolates from this study were sensitive to quinolones. Although the level of resistance to commercial antibiotics was generally low, the use of tetracycline and erythromycin was not recommended.

Keywords: antibiogram, genotyping, Pasteurella multocida, serogrouping, toxA

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Authors: Afroza Khatun, Masuma, Md. Younus Mia, Kamal Kanta Das

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Analysis of the microbial status and physicochemical parameters of some branded and street vended ice cream showed that total viable bacteria in branded ice cream ranged from 4.8×10³ to 1.10×10⁵ cfu/ml, and in street vended ice-cream ranged from 7.5×10⁴ to 1.6×10⁸ cfu/ml. Total coliform bacteria present up to 9.20×10³ cfu/ml in branded ice cream and 5.3×10³ to 9.6×10⁶ cfu/ml observed in street vended ice cream. Total E. coli were found to be present within a range from 0 to 4.5×10³ cfu/ml in branded and 4.1×10² to 7.5×10⁴ cfu/ml in street ice cream. The ranges of Staphylococcus aureus count were 1.8×10² to 2.9×10⁴ cfu/ml (branded) and 3.9×10⁴ to 7.9×10⁶ cfu/ml (street). The pH of both types of ice cream showed acidic to neutral conditions where the concentration of pH for branded ice cream was 5.5 to 6.9, as well as the value of pH in street ice cream, was 6.2 to 7.0. The range of Total soluble solids in several branded ice creams was 26 to 29%, and the value of TSS obtained in street-vended ice-creams ranged from 5 to 10%. The overall results of this research demonstrated that the microbial quality in all street ice creams exceeded the BSTI standard and exhibited lower quality than the industrially produced branded ice creams due to comparatively faulty manufacturing processes and poor hygiene practices. The presence of pathogenic microbes was also observed in branded ice creams which was quite alarming for public health. So it is suggested that the government authorized organization should conduct the proper monitoring system to ensure that both branded and street vended ice-creams are microbiologically safe to prevent public health hazards.

Keywords: food safety, microbiological analysis, physicochemical, ice-cream, E. coli, Staphylococcus aureus

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Authors: V. Baweja, K. K. Gupta, V. Dubey, C. Keshavam

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Antimicrobial activity of six indigenous plants such as Tulsi Ocimum sanctum, Neem Azadirachta indica, Aloe vera, Turmeric Curcuma longa, Lantana Lantana camara, and Clove Syzygium aromaticum was assessed against the gut microbiota of the dengue fever mosquito Aedes aegypti, keeping in view that the presence of midgut bacteria may affect the ability of the vector to transmit pathogens. Eleven different types of bacterial clones were isolated from the midgut of lab-reared fourth instar larvae of Aedes aegypti and were grown on LB agar medium at an optimum temperature of 25 ºC. Identification of these bacteria was done on the basis of their colony characteristic such as colony size, shape, opacity, elevation, consistency, and growth. Light microscopic studies of the gut microbiota revealed dominance of Gram-negative cocci over gram positive cocci and bacilli and Gram-negative bacilli. Identification of species was done by chemical characterization of the colonies. Crude extracts of all test plants were screened for their antimicrobial activities against gut microbiota by disc diffusion assay. The zone of exclusion seen after 24 hr of incubation in different assays revealed the most potent antibacterial activities in neem followed by clove and turmeric. Lantana and Aloe vera were least effective.

Keywords: plant extract, aedes, dengue, antimicrobial activity

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Authors: Allan Kiptanui Kimisto, Geoffrey Odhiambo Ongondo, Anastasia Wairimu Muia, Cyrus Ndungu Kimani

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The major challenge to proper sewage sludge treatment processes is the poor understanding of sludge microbiome diversities. This study applied the whole-genome. shotgun metagenomics technique to profile the microbial composition of sewage sludge in two active digestion lagoons at the Nyeri-Kangemi Wastewater Treatment Plant in Nyeri County, Kenya. Total microbial community DNA was extracted from samples using the available ZymoBIOMICS™ DNA Miniprep Kit and sequenced using Shotgun metagenomics. Samples were analyzed using MG-RAST software (Project ID: mgp100988), which allowed for comparing taxonomic diversity before β-diversities studies for Bacteria, Archaea and Eukaryotes. The study identified 57 phyla, 145 classes, 301 orders, 506 families, 963 genera, and 1980 species. Bacteria dominated the microbes and comprised 28 species, 51 classes, 110 orders, 243 families, 597 genera, and 1518 species. The Bacteroides(6.77%) were dominant, followed by Acinetobacter(1.44%) belonging to the Gammaproteobacteria and Acidororax (1.36%), Bacillus (1.24%) and Clostridium (1.02%) belonging to Betaproteobacteria. Archaea recorded 5 phyla, 13 classes, 19 orders, 29 families, 60 genera,and87 species, with the dominant genera being Methanospirillum (16.01%), methanosarcina (15.70%), and Methanoregula(14.80%) and Methanosaeta (8.74%), Methanosphaerula(5.48%) and Methanobrevibacter(5.03%) being the subdominant group. The eukaryotes were the least in abundance and comprised 24 phyla, 81 classes, 301 orders, 506 families, 963 genera, and 980 species. Arabidopsis (4.91%) and Caenorhabditis (4.81%) dominated the eukaryotes, while Dityostelium (3.63%) and Drosophila(2.08%) were the subdominant genera. All these microbes play distinct roles in the anaerobic treatment process of sewage sludge. The local sludge microbial composition and abundance variations may be due to age difference differences between the two digestion lagoons in operation at the plant and the different degradation rales played by the taxa. The information presented in this study can help in the genetic manipulation or formulation of optimal microbial ratios to improve their effectiveness in sewage sludge treatment. This study recommends further research on how the different taxa respond to environmental changes over time and space.

Keywords: shotgun metagenomics, sludge, bacteria, archaea, eukaryotes

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Authors: Livinus A. Obasi, Augustine N. Ajah

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Biotechnology in recent times has tried to develop a mechanism whereby sustainable electricity can be generated by the activity of microorganisms on waste and renewable biomass (often regarded as “negative value”) in a device called microbial fuel cell, MFC. In this paper, we established how the biocatalytic activities of bacteria on organic matter (substrates) produced some electrons with the associated removal of some water pollution parameters; Biochemical oxygen demand (BOD), chemical oxygen demand (COD) to the tune of 77.2% and 88.3% respectively from a petrochemical sanitary wastewater. The electricity generation was possible by conditioning the bacteria to operate anaerobically in one chamber referred to as the anode while the electrons are transferred to the fully aerated counter chamber containing the cathode. Power densities ranging from 12.83 mW/m² to 966.66 mW/m² were achieved using a dual-chamber starch membrane MFC experimental set-up. The maximum power density obtained in this research shows an improvement in the use of low cost MFC set up to achieve power production. Also, the level of organic matter removal from the sanitary waste water by the operation of this device clearly demonstrates its potential benefit in achieving an improved benign environment. The beauty of the MFCs is their potential utility in areas lacking electrical infrastructures like in most developing countries.

Keywords: bioelectricity, COD, microbial fuel cell, sanitary wastewater, wheat starch

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Authors: Mark Andrew Tu, Raymond Olazo, Cybele Abad

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Cryptococcosis is an invasive infection most commonly found in patients who are immuno compromised. However, patients with this infection usually present with meningitis and rarely pulmonary infection in isolation. We present a case of a Filipino HIV patient who developed cryptococcal pneumonia without meningitis.

Keywords: Cryptococcal Pneumonia, HIV, Filipino, immune system

Procedia PDF Downloads 441