Search results for: Escherichia%20coli%20SCC1

Authors: Bilal Murtaza, Xiaoyu Li, Liming Dong, Muhammad Kashif Saleemi, Gen Li, Bowen Jin, Lili Wang, Yongping Xu

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Fusarium spp. produce numerous mycotoxins, such as zearalenone (ZEN), deoxynivalenol (DON), and its acetylated compounds, 3-acetyl-deoxynivalenol (3-ADON) and 15-acetyl-deoxynivalenol (15-ADON) (15-ADON). In a co-culture system, the soil-derived Escherichia marmotae strain degrades ZEN and DON into 3-keto-DON and DOM-1 via enzymatic deep-oxidation. When pure mycotoxins were subjected to Escherichia marmotae in culture flasks, degradation, and detoxification were also attained. DON and ZEN concentrations, ambient pH, incubation temperatures, bacterium concentrations, and the impact of acid treatment on degradation were all evaluated. The results of the ELISA and high-performance liquid chromatography-electrospray ionization-high resolution mass spectrometry (HPLC-ESI-HRMS) tests demonstrated that the concentration of mycotoxins exposed to Escherichia marmotae was significantly lower than the control. ZEN levels were reduced by 43.9%, while zearalenone sulfate ([M/z 397.1052 C18H21O8S1) was discovered as a derivative of ZEN converted by microbes to a less toxic molecule. Furthermore, Escherichia marmotae appeared to metabolize DON 35.10% into less toxic derivatives (DOM-1 at m/z 281 of [DON - O]+ and 3-keto-DON at m/z 295 of [DON - 2H]+). These results show that Escherichia marmotae can reduce Fusarium mycotoxins production, degrade pure mycotoxins, and convert them to less harmful compounds, opening up new possibilities for study and innovation in mycotoxin detoxification.

Keywords: mycotoxins, zearalenone, deoxynivalenol, bacterial degradation

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Authors: Sana Mohammad Jafar, Hossaini Seyahi Zohreh

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Infections and food intoxication caused by microbial contamination of food is of major issues in different countries, and diseases caused by the consumption of contaminated food included a large percentage of the country's health problems. Since traditional cheese for cultural reasons, good taste and smell in many parts of the area still has the important place in people's food basket, transmission of pathogenic bacteria could be at risk human health through the consumption of this food. In this study selected randomly 100 samples of 250 grams of traditional cheeses supplied in the city Behbahan market and adjacent to the ice was transferred to the laboratory and microbiological tests were performed immediately. According to the results, from 100 samples tested traditional cheese, 94 samples (94% of samples) were contaminated with coliforms, which of this number 75 samples (75% of samples) the contamination rate was higher than the limit (more than 100 cfu/g). Of the total samples, 36 samples (36% of samples) were contaminated with fecal coliform which of this number 30 samples (30% of samples) were contaminated with Escherichia.coli bacteria. Based on the results of agglutination test,no samples was found positive as pathogenic Escherichia.coli.

Keywords: determination, traditional cheese, Behbahan, Escherichia coli

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Authors: Fuji Astuti, Helen Onyeaka

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The aim of the study was to investigate the combine effects of cinnamaldehyde (0.25 and 0.45% v/v) on thermal resistance of pathogenic Escherichia coli during low temperature long time (LT-LT) cooking below 60℃. Three different static temperatures (48, 53 and 50℃) were performed, and the number of viable cells was studied. The starting concentrations of cells were 10⁸ CFU/ml. In this experiment, heat treatment efficiency for safe reduction indicated by decimal logarithm reduction of viable recovered cells, which was monitored for heating over 6 hours. Thermal inactivation was measured by means of establishing the death curves between the mean log surviving cells (log₁₀ CFU/ml) and designated time points (minutes) for each temperature test. The findings depicted that addition of cinnamaldehyde exhibited to elevate the thermal sensitivity of E. coli. However, the injured cells found to be well-adapted to all temperature tests after certain time point of cooking, in which they grew to more than 10⁵ CFU/ml.

Keywords: cinnamaldehyde, decimal logarithm reduction, Escherichia coli, LT-LT cooking

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Authors: S. Techaoei, K. Jarmkom, P. Eakwaropas, W. Khobjai

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The objective of this research was focused on investigating in vitro antimicrobial activity of Phellinus linteus fruiting body extracts on Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. Phellinus linteus fruiting body was extracted with ethanol and ethyl acetate and was vaporized. The disc diffusion assay was used to assess antimicrobial activity against tested bacterial strains. Primary screening of chemical profile of crude extract was determined by using thin layer chromatography. The positive control and the negative control were used as erythromycin and dimethyl sulfoxide, respectively. Initial screening of Phellinus linteus crude extract with the disc diffusion assay demonstrated that only ethanol had greater antimicrobial activity against Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. The MIC assay showed that the lower MIC was observed with 0.5 mg/ml of Pseudomonas aeruginosa and Methicillin-resistant Staphylococcus aureus and 0.25 mg/ml. of Escherichia coli and Staphylococcus aureus, respectively. TLC chemical profile of extract was represented at R_f ≈ 0.71-0.76.

Keywords: Staphylococcus aureus, Escherichia coli, Phellinus linteus, Methicillin-resistant Staphylococcus aureus, antimicrobial activity

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Authors: Bo Ram Choi, Ji Su Kim, Juyeon Cho, Hyukjin Lee

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Food contaminated by bacteria (E.coli), causes food poisoning, which occurs to many patients worldwide annually. We have introduced an application of rolling circle amplification (RCA) as a versatile biosensor and developed a diagnostic platform composed of capillary tube and microbeads for rapid and easy detection of Escherichia coli (E. coli). When specific mRNA of E.coli is extracted from cell lysis, rolling circle amplification (RCA) of DNA template can be achieved and can be visualized by beads aggregation in capillary tube. In contrast, if there is no bacterial pathogen in sample, no beads aggregation can be seen. This assay is possible to detect visually target gene without specific equipment. It is likely to the development of a genetic kit for point of care testing (POCT) that can detect target gene using microbeads.

Keywords: rolling circle amplification (RCA), Escherichia coli (E. coli), point of care testing (POCT), beads aggregation, capillary tube

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Authors: Rehan Deshmukh, Sunil Bhand, Utpal Roy

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We report the label-free electrochemical detection of Escherichia coli O157:H7 (ATCC 43895) in potable water using a DNA probe as a sensing molecule targeting the open reading frame marker. Indium tin oxide (ITO) surface was modified with organosilane and, glutaraldehyde was applied as a linker to fabricate the DNA sensor chip. Non-Faradic electrochemical impedance spectroscopy (EIS) behavior was investigated at each step of sensor fabrication using cyclic voltammetry, impedance, phase, relative permittivity, capacitance, and admittance. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) revealed significant changes in surface topographies of DNA sensor chip fabrication. The decrease in the percentage of pinholes from 2.05 (Bare ITO) to 1.46 (after DNA hybridization) suggested the capacitive behavior of the DNA sensor chip. The results of non-Faradic EIS studies of DNA sensor chip showed a systematic declining trend of the capacitance as well as the relative permittivity upon DNA hybridization. DNA sensor chip exhibited linearity in 0.5 to 25 pg/10mL for E. coli O157:H7 (ATCC 43895). The limit of detection (LOD) at 95% confidence estimated by logistic regression was 0.1 pg DNA/10mL of E. coli O157:H7 (equivalent to 13.67 CFU/10mL) with a p-value of 0.0237. Moreover, the fabricated DNA sensor chip used for detection of E. coli O157:H7 showed no significant cross-reactivity with closely and distantly related bacteria such as Escherichia coli MTCC 3221, Escherichia coli O78:H11 MTCC 723 and Bacillus subtilis MTCC 736. Consequently, the results obtained in our study demonstrated the possible application of developed DNA sensor chips for E. coli O157:H7 ATCC 43895 in real water samples as well.

Keywords: capacitance, DNA sensor, Escherichia coli O157:H7, open reading frame marker

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Authors: O. P. Yadav

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Ag-N co-doped ZnS and ZnS/Fe2O3 composite nanoparticles have been synthesized by chemical and sol-gel methods. As-synthesized nanomaterial have been characterized by XRD and TEM techniques and their antimicrobial effects were studied using paper disc diffusion technique against gram positive (Staphylococcus aureus) and gram negative (Escherichia coli) bacteria. As-synthesized nanomaterial showed potent antimicrobial activity against studied bacterial strains. Antimicrobial activity of synthesized nanomaterial has also been compared with some commonly used antibiotics.

Keywords: antibiotic, Escherichia coli, nanomaterial, TEM, Staphylococcus aureus

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Authors: B. Navidshad

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The main objective of this study was to examine the effects of enzymatic treatment and shell content of palm kernel expeller (PKE) on the ileal Lactobacilli and Escherichia coli populations in broiler chickens. At the finisher phase, one hundred male broiler chickens (Cobb-500) were fed a control diet or the diets containing 200 g/kg of normal PKE (70 g/kg shell), low shell PKE (30 g/kg shell), enzymatic treated PKE or low shell-enzymatic treated PKE. The quantitative real-time PCR were used to determine the ileal bacteria populations. The lowest ileal Lactobacilli population was found in the chickens fed the low shell PKE diet. Dietary normal PKE or low shell-enzymatic treated PKE decreased the Escherichia coli population compared to the control diet. The results suggested that PKE could be included up to 200 g/kg in the finisher diet, however, any screening practice to reduce the shell content of PKE without enzymatic degradation of β-mannan, decrease ileal Lactobacilli population.

Keywords: palm kernel expeller, exogenous enzyme, shell content, ileum bacteria, broiler chickens

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Authors: P. Pasomboon, P. Chumnanpuen, T. E-kobon

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Hyaluronic acid (HA) consists of linear heteropolysaccharides repeat of D-glucuronic acid and N-acetyl-D-glucosamine. HA has various useful properties to maintain skin elasticity and moisture, reduce inflammation, and lubricate the movement of various body parts without causing immunogenic allergy. HA can be found in several animal tissues as well as in the capsule component of some bacteria including Pasteurella multocida. This study aimed to modify a genome-scale metabolic model of Escherichia coli using computational simulation and flux analysis methods to predict HA productivity under different carbon sources and nitrogen supplement by the addition of two enzymes (GLMU2 and HYAD) from P. multocida to improve the HA production under the specified amount of carbon sources and nitrogen supplements. Result revealed that threonine and aspartate supplement raised the HA production by 12.186%. Our analyses proposed the genome-scale metabolic model is useful for improving the HA production and narrows the number of conditions to be tested further.

Keywords: Pasteurella multocida, Escherichia coli, hyaluronic acid, genome-scale metabolic model, bioinformatics

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Authors: Emmanuel Jaluchimike Iloputaife, Kelechi Nkechinyere Mbah-Omeje

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This study was designed to determine the antimicrobial activities and minimum inhibitory concentration of three different batches (Fresh, Oven dried and Sundried) of Ass Hay extracted with water, ethanol and methanolagainst selected human pathogenic microorganisms (Escherichia coli, Klebsiella Pneumonia, Staphylococcus aureus, Aspergillus niger and Candidaalbicans). All extracts were reconstituted with peptone water and tested for antimicrobial activity. The antimicrobial activity, the Minimum Inhibitory Concentration and Minimum Bactericidal/Fungicidal concentrations were determined by agar well diffusion methodagainst test organismsin which aseptic conditions were observed. The antimicrobial activities of the different batches of Ass Hay on the test organisms varied considerably. The highest inhibition zone diameter at 200 mg/ml for the different batches of Ass Hay was recorded by sundried methanol extract against Escherichia coli at 36.4 ± 0.2 mm while fresh methanol extract inhibited Klebsiela pneumonia with the least inhibition zone diameter at 20.1 ± 0.1mm. At 100 mg/ml the highest inhibition zone diameter was recorded by oven dried water extract against Escherichia coli at 30.3 ± 0.3 mm while sun dried water extract inhibited Staphylococcus aureus with the least inhibition zone diameter at 15.1 ± 0.1 mm. At 50mg/ml, the highest inhibition zone diameter was recorded by fresh water extract against Escherichia coli at 25.9 ± 0.1 mm while oven dried water extract inhibited Klebsiela pneumonia with least inhibition zone diameter at 12.1 ± 0.2 mm. At 25mg/ml, the highest inhibition zone diameter was recorded by fresh water extract against Escherichia coli at 18.3 ± 0.2 mm while sun dried ethanol extract inhibited Escherichia coli with least inhibition zone diameter at 10.1 ± 0.1 mm. The MIC and MBC result of ethanol extract of fresh Ass Hay showed a uniform value of 6.25 mg/ml and 12.5 mg/ml respectively for all test bacterial isolates. The Minimum Inhibitory concentration and Minimum bactericidal concentration results of Oven dried ethanol Ass Hay extract showed a uniform value of 3.125 mg/ml and 6.25 mg/ml respectively for all test bacterial isolates and Minimum fungicidal concentration value of 12.5 mg/ml for Aspergillus niger. Statistical analysis showed there is significant difference in mean zone inhibition diameter of the products at p < 0.05, p = 0.019. This study has shown there is antimicrobial potential in Ass Hay and at such there is need to further exploit Donkey Ass Hay in order to maximize the potential.

Keywords: microorganisms, Ass Hay, antimicrobial activity, extracts

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Authors: Zoran Herceg, Višnja Stulić, Anet Režek Jambrak, Tomislava Vukušić

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Electrical discharge plasma is a new non-thermal processing technique which is used for the inactivation of contaminating and hazardous microbes in liquids. Plasma is a source of different antimicrobial species including UV photons, charged particles, and reactive species such as superoxide, hydroxyl radicals, nitric oxide and ozone. Escherichia coli was studied as foodborne pathogen. The aim of this work was to examine inactivation effects of electrical discharge plasma treatment on the Escherichia coli MG 1655 in pure culture. Two types of plasma configuration and polarity were used. First configuration was with titanium wire as high voltage needle and another with medical stainless steel needle used to form bubbles in treated volume and titanium wire as high voltage needle. Model solution samples were inoculated with Escerichia coli MG 1655 and treated by electrical discharge plasma at treatment time of 5 and 10 min, and frequency of 60, 90 and 120 Hz. With the first configuration after 5 minutes of treatment at frequency of 120 Hz the inactivation rate was 1.3 log₁₀ reduction and after 10 minutes of treatment the inactivation rate was 3.0 log₁₀ reduction. At the frequency of 90 Hz after 10 minutes inactivation rate was 1.3 log₁₀ reduction. With the second configuration after 5 minutes of treatment at frequency of 120 Hz the inactivation rate was 1.2 log₁₀ reduction and after 10 minutes of treatment the inactivation rate was also 3.0 log₁₀ reduction. In this work it was also examined the formation of biofilm, nucleotide and protein leakage at 260/280 nm, before and after treatment and recuperation of treated samples. Further optimization of method is needed to understand mechanism of inactivation.

Keywords: electrical discharge plasma, escherichia coli MG 1655, inactivation, point-to-plate electrode configuration

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Authors: Debjyoti Bhakat, Indranil Mondal, Asish K. Mukhopadayay, Nabendu S. Chatterjee

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Enterotoxigenic Escherichia coli is one of the leading causes of diarrhea in infants and travelers in developing countries. Colonization factors play an important role in pathogenesis and are one of the main targets for Enterotoxigenic Escherichia coli (ETEC) vaccine development. However, ETEC vaccines had poorly performed in the past, as the prevalence of colonization factors is region-dependent. There are more than 25 classical colonization factors presently known to be expressed by ETEC, although all are not expressed together. Further, there are other multiple non-classical virulence factors that are also identified. Here the presence and expression of common classical and non-classical virulence factors were studied. Further studies were done on the expression of prevalent colonization factors in different strains. For the prevalence determination, multiplex polymerase chain reaction (PCR) was employed, which was confirmed by simplex PCR. Quantitative RT-PCR was done to study the RNA expression of these virulence factors. Strains negative for colonization factors expression were confirmed by SDS-PAGE. Among the clinical isolates, the most prevalent toxin was est+elt, followed by est and elt, while the pattern was reversed in the control strains. There were 29% and 40% strains negative for any classical colonization factors (CF) or non-classical virulence factors (NCVF) among the clinical and control strains, respectively. Among CF positive ETEC strains, CS6 and CS21 were the prevalent ones in the clinical strains, whereas in control strains, CS6 was the predominant one. For NCVF genes, eatA was the most prevalent among the clinical isolates and etpA for control. CS6 was the most expressed CF, and eatA was the predominantly expressed NCVF for both clinical and controlled ETEC isolates. CS6 expression was more in strains having CS6 alone. Different strains express CS6 at different levels. Not all strains expressed their respective virulence factors. Understanding the prevalent colonization factor, CS6, and its nature of expression will contribute to designing an effective vaccine against ETEC in this region of the globe. The expression pattern of CS6 also will help in examining the relatedness between the ETEC subtypes.

Keywords: classical virulence factors, CS6, diarrhea, enterotoxigenic escherichia coli, expression, non-classical virulence factors

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Authors: Raana Babadi Fathipour

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Ice cream is a nutritious dairy product that, given its constituent materials and high nutritional value, is a suitable growth medium for the growth of various food microorganisms. The contamination of this product with pathogenic microorganisms may cause food poisoning and infections, and so could be harmful to human health. The foremost critical pathogenic microscopic organisms of ice cream incorporate Escherichia coli, Staphylococcus aureus, Bacillus cereus, Enterobacteriaceae, coliforms, Listeria monocytogenes and Enterococcus. Biosensor technology, albeit a recent addition to the dairy industry, has proven its worth in other fields, such as medical devices. Through numerous studies, the advantages of employing biosensors have consistently emerged. These incredible tools present expeditious and straightforward means while specifically targeting analytes. Thus, they bring forth unparalleled solutions that bolster ongoing advancements within dairy products and processes. This review delves into the latest developments in the realm of biosensors and evaluates the diverse techniques of bio-recognition and transduction in terms of their benefits, drawbacks, and relevance to traditional ice cream. Furthermore, the obstacles that impede the progress of these approaches in meeting the growing need for swift and real-time quality control of milk products, particularly ice cream, are also expounded upon.

Keywords: traditional ice cream, Escherichia coli, Staphylococcus aureus, biosensors

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Authors: Desouky Abd El Haleem, Sahar Zaki

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This work was conducted to develop a one-step multiplex PCR system for rapid, sensitive, and specific detection of three different bacterial pathogens, Escherichia coli, Pseudomonas aeruginosa, and Salmonella spp, directly in wastewater without prior isolation on selective media. As a molecular confirmatory test after isolation of the pathogens by classical microbiological methods, PCR-RFLP of their amplified 16S rDNA genes was performed. It was observed that the developed protocols have significance impact in the ability to detect sensitively, rapidly and specifically the three pathogens directly in water within short-time, represents a considerable advancement over more time-consuming and less-sensitive methods for identification and characterization of these kinds of pathogens.

Keywords: multiplex PCR, bacterial pathogens, Escherichia coli, Pseudomonas aeruginosa, Salmonella spp.

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Authors: I. N. Nwafia, U. C. Ozumba, M. E. Ohanu, S. O. Ebede

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Antibiotic resistance is increasing globally and has become a major health challenge. Extended-spectrum beta-lactamase is clinically important because the ESBL gene are mostly plasmid encoded and these plasmids frequently carry genes encoding resistance to other classes of antimicrobials thereby limiting antibiotic options in the treatment of infections caused by these organisms. The specific objectives of this study were to determine the prevalence of ESBLs production in Escherichia coli, to determine the antibiotic susceptibility pattern of ESBLs producing Escherichia coli, to detect TEM, SHV and CTX-M genes and the risk factors to acquisition of ESBL producing Escherichia coli. The protocol of the study was approved by Health Research and Ethics committee of the University of Nigeria Teaching Hospital (UNTH), Enugu. It was a descriptive cross-sectional study that involved all hospitalized patients in UNTH from whose specimens Escherichia coli was isolated during the period of the study. The samples analysed were urine, wound swabs, blood and cerebrospinal fluid. These samples were cultured in 5% sheep Blood agar and MacConkey agar (Oxoid Laboratories, Cambridge UK) and incubated at 35-370C for 24 hours. Escherichia coli was identified with standard biochemical tests and confirmed using API 20E auxanogram (bioMerieux, Marcy 1'Etoile, France). The antibiotic susceptibility testing was done by disc diffusion method and interpreted according to the Clinical and Laboratory Standard Institute guideline. ESBL production was confirmed using ESBL Epsilometer test strips (Liofilchem srl, Italy). The ESBL bla genes were detected with polymerase chain reaction, after extraction of DNA with plasmid mini-prep kit (Jena Bioscience, Jena, Germany). Data analysis was with appropriate descriptive and inferential statistics. One hundred and six isolates (53.00%) out of the 200 were from urine, followed by isolates from different swabs specimens 53(26.50%) and the least number of the isolates 4(2.00) were from blood (P value = 0.096). Seventy (35.00%) out of the 200 isolates, were confirmed positive for ESBL production. Forty-two (60.00%) of the isolates were from female patients while 28(40.00%) were from male patients (P value = 0.13). Sixty-eight (97.14%) of the isolates were susceptible to imipenem while all of the isolates were resistant to ampicillin, chloramphenicol and tetracycline. From the 70 positive isolates the ESBL genes detected with polymerase chain reaction were blaCTX-M (n=26; 37.14%), blaTEM (n=7; 10.00%), blaSHV (n=2; 2.86%), blaCTX-M/TEM (n=7; 10.0%), blaCTX-M/SHV (n=14; 20.0%) and blaCTX-M/TEM/SHV (n=10; 14.29%). There was no gene detected in 4(5.71%) of the isolates. The most associated risk factors to infections caused by ESBL producing Escherichia coli was previous antibiotics use for the past 3 months followed by admission in the intensive care unit, recent surgery, and urinary catheterization. In conclusion, ESBLs was detected in 4 of every 10 Escherichia coli with the predominant gene detected being CTX-M. This knowledge will enable appropriate measures towards improvement of patient health care, antibiotic stewardship, research and infection control in the hospital.

Keywords: antimicrobial, Escherichia coli, extended spectrum beta lactamase, resistance

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Authors: Refal Hussain, Saifuddin M. Nomanbhay

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Glycerol is a valuable raw material for the production of industrially useful metabolites. Among many promising applications for the use of glycerol is its bioconversion to high value-added compounds, such as bioethanol through microbial fermentation. Bioethanol is an important industrial chemical with emerging potential as a biofuel to replace vanishing fossil fuels. The yield of liquid fuel in this process was greatly influenced by various parameters viz, temperature, pH, glycerol concentration, organic concentration, and agitation speed were considered. The present study was undertaken to investigate optimum parameters for bioethanol production from raw glycerol by immobilized mutant Escherichia coli (E.coli) (ATCC11505) strain on chitosan cross linked glutaraldehyde optimized by Taguchi statistical method in shake flasks. The initial parameters were set each at four levels and the orthogonal array layout of L16 (45) conducted. The important controlling parameters for optimized the operational fermentation was temperature 38 °C, medium pH 6.5, initial glycerol concentration (250 g/l), and organic source concentration (5 g/l). Fermentation with optimized parameters was carried out in a custom fabricated shake flask. The predicted value of bioethanol production under optimized conditions was (118.13 g/l). Immobilized cells are mainly used for economic benefits of continuous production or repeated use in continuous as well as in batch mode.

Keywords: bioethanol, Escherichia coli, immobilization, optimization

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Authors: Nadia A. Mohamed, Magdy W. Sabaa, Ahmed H. H. El-Ghandour, Marwa M. Abdel-Aziz, Omayma F. Abdel-Gawad

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Eighteen carboxymethyl chitosan (CMCh) schiff bases and their reduced derivatives have been synthesized. They were characterized by spectral analyses (FT-IR and H1-NMR) and scanning electron microscopy observation. Their antibacterial activities against Streptococcus pneumoniae (RCMB 010010), Bacillis subtilis (RCMB 010067), as Gram positive bacteria and Escherichia coli (RCMB 010052) as Gram negative bacteria and the antifungal activity against Aspergillus fumigatus (RCMB 02568), Geotricum candidum (RCMB 05097), and Candida albicans (RCMB 05031) were examined using agar disk diffusion method. The results demonstrate how the antibacterial and the antifungal activity are clearly affected by both the nature and position of the substituent groups in the aryl ring of the prepared derivatives. CMCh-4-nitroBenz Schiff base and its reduced form show higher antimicrobial activity comparing with other para substituted derivatives. CMCh-4-nitroBenz Schiff base: 18.3, 17, and 15.6 mm against Bacillis subtilis, Streptococcus pneumonia, and Escherichia coli respectively and 16.2, 17.3, and 16.4 mm against Aspergillus fumigates, Geotricum candidum, and Candida albicans respectively. CMCh-4-nitroBenz reduced form: 19.5, 18.7, and 16.2 mm against Bacillis subtilis, Streptococcus pneumonia, and Escherichia coli respectively and 17.5, 19.5, and 17.4 mm against Aspergillus fumigates, Geotricum candidum, and Candida albicans respectively. Also CMCh-3-bromoBenz show good results; CMCh-3-bromoBenz schiff base: 19.2, 16.9, and 14.6 mm Bacillis subtilis, Streptococcus pneumonia, and Escherichia coli respectively and 18.4, 17.6, and 15.9 mm against Aspergillus fumigates, Geotricum candidum, and Candida albicans respectively.

Keywords: chitosan, schiff base, minimum inhibition concentration, antimicrobial activity

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Authors: Pui Shan Wong, Kosuke Tashiro, Satoru Kuhara, Sachiyo Aburatani

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Functional genomics and gene regulation inference has readily expanded our knowledge and understanding of gene interactions with regards to expression regulation. With the advancement of transcriptome sequencing in time-series comes the ability to study the sequential changes of the transcriptome. This method presented here works to augment existing regulation networks accumulated in literature with transcriptome data gathered from time-series experiments to construct a sequential representation of transcription factor activity. This method is applied on a time-series RNA-Seq data set from Escherichia coli as it transitions from growth to stationary phase over five hours. Investigations are conducted on the various metabolic activities in gene regulation processes by taking advantage of the correlation between regulatory gene pairs to examine their activity on a dynamic network. Especially, the changes in metabolic activity during phase transition are analyzed with focus on the pagP gene as well as other associated transcription factors. The visualization of the sequential transcriptional activity is used to describe the change in metabolic pathway activity originating from the pagP transcription factor, phoP. The results show a shift from amino acid and nucleic acid metabolism, to energy metabolism during the transition to stationary phase in E. coli.

Keywords: Escherichia coli, gene regulation, network, time-series

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Authors: Sanja O. Podunavac-Kuzmanović, Lidija R. Jevrić, Strahinja Z. Kovačević

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In last decade, metal-organic complexes have captured intensive attention because of their wide range of biological activities such as antibacterial, antifungal, anticancerous, antimicrobial and antiHIV. Therefore, it is of great importance for the development of coordination chemistry to explore the assembly of functional organic ligands with metal ion and to investigate the relationship between the structure and property. In view of our studies, we reasoned that benzimidazoles complexed to metal ions could act as a potent antibacterial agents. Thus, we have bioassayed the inhibitory potency of benzimidazoles and their metal salts (Co or Ni) against Gram negative bacteria Escherichia coli. In order to validate our in vitro study, we performed in silico studies using molecular docking software’s. The investigated compounds and their metal complexes (Co, Ni) showed good antibacterial activity against Escherichia coli. In silico docking studies of the synthesized compounds suggested that complexed benzimidazoles have a greater binding affinity and enhanced antibacterial activity in comparison with noncomplexed ligands. In view of their enhanced inhibitory properties we propose that the studied complexes can be used as potential pharmaceuticals. This study is financially supported by COST action CM1306 and the project No. 114-451-347/2015-02, financially supported by the Provincial Secretariat for Science and Technological Development of Vojvodina.

Keywords: benzimidazoles, complexes, antibacterial, Escherichia coli, metal

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Authors: Fariha Akhter Chowdhury, Mohammad Nurul Islam, Anamika Saha, Sabrina Mahboob, Abu Syed Md. Mosaddek, Md. Omar Faruque, Most. Fahmida Begum, Rajib Bhattacharjee

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Urinary tract infection (UTI) is one of the most common infectious diseases in Bangladesh where Escherichia coli is the prevalent organism and responsible for most of the infections. Lipopolysaccharide (LPS) is known to act as a major virulence factor of E. coli. The present study aimed to purify, extract and visualize LPS of E. coli clinical isolates from urine samples of patients with UTI. The E. coli strain was isolated from the urine samples of 10 patients with UTI and then the antibiotic sensitivity pattern of the isolates was determined. The purification of LPS was carried out using the hot aqueous-phenol method and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, which was directly stained using the modified silver staining method and Coomassie blue. The silver-stained gel demonstrated both smooth and rough type LPS by showing trail-like band patterns with the presence and lacking O-antigen region, respectively. Coomassie blue staining showed no band assuring the absence of any contaminating protein. Our successful extraction of purified LPS from E. coli isolates of UTI patients’ urine samples can be an important step to understand the UTI disease conditions.

Keywords: Escherichia coli, electrophoresis, polyacrylamide gel, silver staining, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)

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Authors: Karima Ould Yerou, B. Meddah, A. Tir Touil

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The intestinal flora also called the intestinal microbiota, consists of different bacteria and other microorganisms which occur naturally in the gastrointestinal tract organs components. These intestinal bacteria are present in their millions and help the functioning of the body in particular allowing aid to degradation of certain molecules into absorbable substrates. They also protect against invasion of the gut by other pathogenic bacteria, that is to say which may be responsible for disease. Factors like stress, antibiotics and diet can affect the balance of intestinal flora and in case of imbalance, digestive disorders type bloating, diarrhea or vomiting may occur. Rattus norvegicus of bad weight of 100 kg, an Algerian firm West are scarified and isolation of their ileum and colon respectively two Bactrian strains Escherichia coli and Lactobacillus are then purified and identified.

Keywords: intestinal flora, Rattus norvegicus, Escherichia coli, lactobacillus, West Algerian farm

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Authors: Abdullahi Musa, Yakubu Kukure Enebe Ibrahim, Adeshina Gujumbola

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The emergence of multidrug resistance in clinical Escherichia coli has been associated with plasmid-mediated genes. DNA transfer among bacteria is critical to the dissemination of resistance. Plasmids have proved to be the ideal vehicles for dissemination of resistance genes. Plasmids coding for antibiotic resistance were long being recognized by many researchers globally. The study aimed at determining the antibiotic susceptibility pattern of ESBL E. coli isolates claimed to be multidrug resistance using disc diffusion method. Antibacterial activity of the test isolates was carried out using disk diffusion methods. The results showed that, majority of the multidrug resistance among clinical isolates of ESBL E. coli was as a result of acquisition of plasmid carrying antibiotic-resistance genes. Production of these ESBL enzymes by these organisms which are normally carried by plasmid and transfer from one bacterium to another has greatly contributed to the rapid spread of antibiotic resistance amongst E. coli isolates, which lead to high economic burden, increase morbidity and mortality rate, complication in therapy and limit treatment options. To curtail these problems, it is of significance to checkmate the rate at which over the counter drugs are sold and antibiotic misused in animal feeds. This will play a very important role in minimizing the spread of resistance bacterial strains in our environment.

Keywords: Escherichia coli, plasmid, multidrug resistance, ESBL, pan drug resistance

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Authors: Tzyy-Rong Jinn, Sheng-Kuo Hsieh, Yi-Ching Chung, Feng-Chia Hsieh

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In this study, we attempted to develop a recombinant oleosin-based fusion expression strategy in Escherichia coli (E. coli) and coupled with the artificial oil bodies (AOB)-cyanogen bromide technology platform to produce bioactive mastoparan B (MP-B). As reported, the oleosin in AOB system plays a carrier (fusion with target protein), since oleosin possess two amphipathic regions (at the N-terminus and C-terminus), which result in the N-terminus and C-terminus of oleosin could be arranged on the surface of AOB. Thus, the target protein fused to the N-terminus or C-terminus of oleosin which also is exposed on the surface of AOB, and this process will greatly facilitate the subsequent separation and purification of target protein from AOB. In addition, oleosin, a unique structural protein of seed oil bodies, has the added advantage of helping the fused MP-B expressed in inclusion bodies, which can protect from proteolytic degradation. In this work, MP-B was fused to the C-terminus of oleosin and then was expressed in E. coli as an insoluble recombinant protein. As a consequence, we successfully developed a reliable recombinant oleosin-based fusion expression strategy in Escherichia coli and coupled with the artificial oil bodies (AOB)-cyanogen bromide technology platform to produce the small peptide, MP-B. Take together, this platform provides an insight into the production of active MP-B, which will facilitate studies and applications of this peptide in the future.

Keywords: artificial oil bodies, Escherichia coli, Oleosin-fusion protein, Mastoparan-B

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Authors: N. Zeinalzadeh, Mahdi Sadeghi

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Enterotoxigenic Escherichia coli (ETEC) is the most common cause of non-inflammatory diarrhea in the developing countries, resulting in approximately 20% of all diarrheal episodes in children in these areas. ST is one of the most important virulence factors and CFA/I is one of the frequent colonization factors that help to process of ETEC infection. ST and CfaB (CFA/I subunit) are among vaccine candidates against ETEC. So, ST because of its small size is not a good immunogenic in the natural form. However to increase its immunogenic potential, here we explored candidate positions for ST insertion in CfaB sequence. After bioinformatics analysis, one of the candidate positions was selected and the chimeric gene (cfaB*st) sequence was synthesized and expressed in E. coli BL21 (DE3). The chimeric recombinant protein was purified with Ni-NTA columns and characterized with western blot analysis. The residue 74-75 of CfaB sequence could be a good candidate position for ST and other epitopes insertion.

Keywords: bioinformatics, CFA/I, enterotoxigenic E. coli, ST toxoid

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Authors: Cheng-Chih Tsai, Yu-Hsuan Liu, Cheng-Ying Ho, Chun-Chin Huang

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The aim of this study evaluated the in vitro and in vivo antimicrobial activity of selected lactic acid bacteria (LAB) against Uropathogenic Escherichia coli (UPEC) for prevention and amelioration of UTIs. We screened LAB strains with antimicrobial effects on UPEC using a well-diffusion assay, bacterial adherence to the uroepithelium cell line SV-HUC-1 (BCRC 60358), and a coculture inhibition assay. The results showed that the 7 LAB strains (Lactobacillus paracasei, L. salivarius, two Pediococcus pentosaceus strains, two L. plantarum strains, and L. crispatus) and the fermented probiotic products produced by these multi-LAB strains exhibited potent zones of inhibition against UPEC. Moreover, the LAB strains and probiotic products adhered strongly to the uroepithelium SV-HUC-1 cell line. The growth of UPEC strains was also markedly inhibited after co-culture with the LAB strains and probiotic products in human urine. In addition, the enhanced levels of IL-6, IL-8 and lactic acid dehydrogenase were significantly decreased by treatments with the LAB strains and probiotic products in UPEC-induced SV-HUC-1 cells. Furthermore, oral administration of probiotic products reduced the number of viable UPEC in the urine of UPEC-challenged BALB/c mice. Taken together, this study demonstrates that probiotic supplementation may be useful as an adjuvant therapy for the treatment of bacterial-induced urinary tract infections.

Keywords: lactic acid bacterium, SV-HUC-1 uroepithelium, urinary tract infection, uropathogenic Escherichia coli, BALB/c mice

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Authors: Siti Helmyati, Euis Nurdiyawati, Joko Susilo, Endri Yuliati, Siti Fadhilatun Nashriyah, Kurnia Widyastuti

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Background: Iron fortification is one of the most effective and sustainable strategies to overcome anemia. It contradictively, has negative effect on gut microbiota balance. Pathogenic bacteria required iron for their growth. The iron source have greatly affect iron absorption in the intestine. Probiotic can inhibit the growth of pathogen. Lactobacillus plantarum Dad 13, Indonesian local isolate provides many benefits for health while fructo-oligosaccharides (FOS) provides selective substrates for probiotics’ growth. Objective: To determine the effect of iron fortification (NaFeEDTA and FeSO4) on antibacterial activity of synbiotic fermented milk. Methods: The antibacterial activity test was performed using the disc diffusion method. Paper discs were soaked in three kinds of synbiotic fermented milk, which are: 1) fortified with NaFeEDTA, 2) FeSO4 and 3) control. Escherichia coli was inoculated on nutrient agar medium. The ability of inhibition was shown by the formation of clear zone around the paper disc and measured in diameter (mm). Results: Synbiotic fermented milk fortified with iron (either NaFeEDTA or FeSO4) had antibacterial activity against Escherichia coli with diameter of clear zone were 6.53 mm and 12.3 mm, respectively (p<0.05). Compared to control (10.73 mm), synbiotic fermented milk fortified with FeSO4 had similar antibacterial activity (p>0.05). Conclusions: In vitro, synbiotic fermented milk fortified with NaFeEDTA and FeSO4 had different antibacterial activity against Escherichia coli. Iron fortification compound affected the antibacterial activity of synbiotic fermented milk.

Keywords: lactobacillus plantarum Dad 13, FOS, NaFeEDTA, FeSO4, antibacterial activity

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Authors: Debjani Ghosh, Goutam Chowdhury, Prosenjit Samanta, Asish Kumar Mukhopadhyay

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Acute diarrhoea caused by diarrheagenic Escherichia coli (DEC) is one of the major public health problem in developing countries, mainly in Asia and Africa. DEC consists of six pathogroups, but the majority of the cases were associated with the three pathogropus, enterotoxigenic E. coli (ETEC), enteroaggregative E. coli (EAEC), and enteropathogenic E. coli (EPEC). Hence, we studied the prevalence and antimicrobial resistance of these three major DEC pathogroups in hospitalized diarrheal patients in Kolkata, India, during 2012-2019 with a large sample size. 8,891 stool samples were processed, and 7.8% of them was identified as DEC infection screened by multiplex PCR, in which ETEC was most common (47.7%) followed by EAEC (38.4%) and EPEC (13.9%). Clinical patient history suggested that children <5 years of age were mostly affected with ETEC and EAEC, whereas people within >5-14 years of age were significantly associated with EPEC and ETEC infections. Antibiogram profile showed a high prevalence of multidrug resistant (MDR) isolates among DEC (56.9%), in which 9% were resistant to antibiotics of six different antimicrobial classes. Screening of the antibiotic resistance conferring genes in DEC showed the presence of blaCTX-M (30.2%) in highest number followed by blaTEM (27.5%), tetB (18%), sul2 (12.6%), strA (11.8%), aadA1 (9.8%), blaOXA-1 (9%), dfrA1 (1.6%) and blaSHV (1.2%) which indicates the existence of mobile genetic elements in those isolates. Therefore, the presence of MDR DEC strains in higher number alarms the public health authorities to take preventive measures before the upsurge of the DEC caused diarrhea cases in near future.

Keywords: diarrheagenic escherichia coli, ETEC, EAEC, EPEC

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Authors: Ali Fadhel Ahmed, Tuqa Abdulkareem Hameed

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

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

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Authors: Charalampos N. Bompas, Eleni Poulou-Sidiropoulou, Martina Samiotaki, Alexios Vlamis-Gardikas

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Ιn all living organisms, antioxidant defenses are orchestrated by the thioredoxin (Trx) and glutaredoxin (Grx) systems. The Trx system of Escherichia coli (E. coli) is comprised of Trx1 and Trx2, both reduced by thioredoxin reductase (TrxR). The Grx system consists of four Grxs (Grx1, Grx2, Grx3, and Grx4), all reduced by glutathione (GSH) except for Grx4, which is reduced by TrxR. Under normal conditions, the GSH reductase of the Grx system keeps GSH at its reduced state. NADPH+ provides the electrons for all reductions in the Trx and Grx systems. Although the role of the E. coli Trx system is widely known, the function of the Grx system reflects the main property of Grx1, which is the reduction of ribonucleotide reductase Ia (RRIa). E. coli Grx3 (encoded by grxC) may also reduce RRIa in vitro but with slow kinetics. The molecule may account for up to 0.4% of total soluble protein and has been the subject of extensive structural studies. Its biological function, however, remains unknown. Herein, affinity chromatography with monothiol Grx3 serving as bait was used to detect the interactions of Grx3 with other proteins. Different types of interactions were identified (covalent, weak, and strong non-covalent) that suggested novel functions for Grx3. In silico approaches were employed to validate selected interactions. In addition, total protein extracts from the null mutant for grxC and the wild-type strain were compared. The overall findings suggest that Grx3 is involved in various metabolic processes, protein synthesis, and stress responses, expanding the recognized functions of Grx3 beyond the possible reduction of RRIa.

Keywords: escherichia coli, glutaredoxin 3, interactome, thiol-disulfide oxidoreductase

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Authors: Naureen Akhtar

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The stability of the protein in detergent-containing solution is the key for its successful crystallisation. Fluorescence-detection size-exclusion chromatography (FSEC) is a potential approach for screening monodispersity as well as the stability of protein in a detergent-containing-solution. In this present study, covalently linked Green Fluorescent Protein (GFP) to bacterial nitrate transporter, NarU from Escherichia coli was studied for pre-crystallisation trials by FSEC. Immobilised metal ion affinity chromatography (IMAC) and gel filtration were employed for their purification. The main objectives of this study were over-expression, detergent screening and crystallisation of nitrate transporter proteins. This study could not produce enough proteins that could realistically be taken forward to achieve the objectives set for this particular research. In future work, different combinations of variables like vectors, tags, creation of mutant proteins, host cells, position of GFP (N- or C-terminal) and/or membrane proteins would be tried to determine the best combination as the principle of technique is still promising.

Keywords: transporters, detergents, over-expression, crystallography

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