Search results for: Escherichia coli bacteria
1679 Characterization of Triterpenoids Antimicrobial Potential in Ethyl Acetate Extracts from Aerial Parts of Deinbollia Pinnata
Authors: Rufai Yakubu And Suleiman Kabiru
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Triterpenoids are a diverse class of secondary metabolites with potential antimicrobial properties. In this study, the crude extracts from ethyl acetate was obtained with ultrasonic extraction method. Using a combined chromatographic separation method to isolate squalene (1) stigmasterol (2), stigmasta-5,22-diene-3-ol acetate (3), γ-sitosterol (4), lupeol (5), taraxasterol (6), and betulinic acid (7) from ethyl acetate extracts. Ethyl acetate crude extracts and isolated compounds were both screened for antimicrobial activity and minimum inhibitory concentration (MIC). For ethyl acetate crude extracts with concentrations of (1.5, 0.75, 0.35, & 0.168 mg/mL) indicated marginal antibacterial activity with a range of 17, 20 and 14 mm zone of inhibition for Staphylococcus aureus, Escherichia coli and Candida albicans and lower minimum inhibitory concentrations ranges from 18.75 µg/ml to 150 µg/mL. Butulinic acid showed the highest activity against E. coli and C. albicans at 15 mm and 15 mm followed by Lupeol against S. aureus, E. coli and C. albicans at 13, 12, 12 mm. Moreso, no antimicrobial activity for both S. aureus and C. albicans with squalene except for E. coli which showed activity at 11 mm with 300 µg/mL (MIC). Thus, abundant triterpenoids in Deinbollia pinnata will be another centered area for antimicrobial drug discovery.Keywords: triterpenoid, antimicrobial potentials, deinbollia pinnata, aerial parts
Procedia PDF Downloads 711678 Bacteria Flora in the Gut and Respiratory Organs of Clarias gariepinus in Fresh and Brackish Water Habitats of Ondo State, South/West Nigeria
Authors: Nelson R. Osungbemiro, Rafiu O. Sanni, Rotimi F. Olaniyan, Abayomi O. Olajuyigbe
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Bacteria flora of Clarias gariepinus collected from two natural habitats namely Owena River (freshwater) and Igbokoda lagoon (brackish water) were examined using standard microbiological procedures. Thirteen bacterial species were identified. The result indicated that from the identified bacteria isolated, Vibrio sp, Proteus sp. Shigella sp. and E. coli were present in both habitats (fresh and brackish waters). Others were habitat-selective such as Salmonella sp., Pseudomonas sp, Enterococcus sp, Staphylococcus sp. that were found only in freshwater habitat. While Branhamella sp, Streptococcus sp. and Micrococcus sp. were found in brackish water habitat. Bacteria load from Owena river (freshwater) was found to be the highest load recorded at 6.21 x 104cfu. T-test analysis also revealed that there was a marked significant difference between bacterial load in guts of sampled Clarias from fresh water and brackish water habitats.Keywords: bacteria flora, gut, Clarias gariepinus, Owena river
Procedia PDF Downloads 4611677 Production and Purification of Salmonella Typhimurium MisL Autotransporter Protein in Escherichia coli
Authors: Neslihan Taskale Karatug, Mustafa Akcelik
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Some literature data show that misL protein play a role on host immune response formed against Salmonella Typhimurium. The aim of the present study is to learn the role of the protein in S. Typhimurium pathogenicity. To describe certain functions of the protein, primarily recombinant misL protein was produced and purified. PCR was performed using a primer set targeted to passenger domain of the misL gene on S. Typhimurium LT2 genome. Amplicon and pet28a vector were enzymatically cleaved with EcoRI and NheI. The digested DNA materials were purified with High Pure PCR Product Purification Kit. The ligation reaction was achieved with the pure products. After preparation of competent Escherichia coli Dh5α, ligation mix was transformed into the cell by electroporation. To confirm the existence of insert gene, recombinant plasmid DNA of Dh5α was isolated with high pure plasmid DNA kit. Proved the correctness of recombinant plasmid was electroporated to BL21. The cell was induced by IPTG. After induction, the presence of recombinant protein was checked by SDS-PAGE. The recombinant misL protein was purified using HisPur Ni-NTA spin colon. The pure protein was shown by SDS-PAGE and western blot immünoassay. The concentration of the protein was measured BCA Protein Assay kit. In the wake of ligation with digested products (2 kb misL and 5.4 kb pet28a) visualised on gel size of the band was about 7.4 kb and was named as pNT01. The pNT01 recombinant plasmid was transformed into Dh5α and colonies were chosen in selective medium. Plasmid DNA isolation from them was carried out. PCR was achieved on the pNT01 to check misL and 2 kb band was observed on the agarose gel. After electroporation of the plasmid and induction of the cell, 68 kDa misL protein was seen. Subsequent to the purification of the protein, only a band was observed on SDS-PAGE. Association of the pure protein with anti-his antibody was verified by the western blot assay. The concentration of the pure misL protein was determined as 345 μg/mL. Production of polyclonal antibody will be achieved by using the obtained pure recombinant misL protein as next step. The role of the protein will come out on the immune system together some assays.Keywords: cloning, Escherichia coli, recombinant protein purification, Salmonella Typhimurium
Procedia PDF Downloads 3921676 Antimicrobial Effect of Essential Oil of Plant Schinus molle on Some Bacteria Pathogens
Authors: Mehani Mouna, Ladjel segni
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Humans use plants for thousands of years to treat various ailments, In many developing countries, Much of the population relies on traditional doctors and their collections of medicinal plants to cure them. Essential oils have many therapeutic properties. In herbal medicine, They are used for their antiseptic properties against infectious diseases of fungal origin, Against dermatophytes, Those of bacterial origin. The aim of our study is to determine the antimicrobial effect of essential oils of the plant Schinus molle on some pathogenic bacteria. It is a medicinal plant used in traditional therapy. Essential oils have many therapeutic properties. In herbal medicine, They are used for their antiseptic properties against infectious diseases of fungal origin, Against dermatophytes, Those of bacterial origin. The test adopted is based on the diffusion method on solid medium (Antibiogram), This method allows to determine the susceptibility or resistance of an organism according to the sample studied. Our study reveals that the essential oil of the plant Schinus molle has a different effect on the resistance of germs: For Pseudomonas aeruginosa strain is a moderately sensitive with an inhibition zone of 10 mm, Further Antirobactere, Escherichia coli and Proteus are strains that represent a high sensitivity, A zone of inhibition equal to 14.66 mm.Keywords: Essential oil, microorganism, antibiogram, shinus molle
Procedia PDF Downloads 3471675 Parametric Study of a Washing Machine to Develop an Energy Efficient Program Regarding the Enhanced Washing Efficiency Index and Micro Organism Removal Performance
Authors: Peli̇n Yilmaz, Gi̇zemnur Yildiz Uysal, Emi̇ne Bi̇rci̇, Berk Özcan, Burak Koca, Ehsan Tuzcuoğlu, Fati̇h Kasap
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Development of Energy Efficient Programs (EEP) is one of the most significant trends in the wet appliance industry of the recent years. Thanks to the EEP, the energy consumption of a washing machine as one of the most energy-consuming home appliances can shrink considerably, while its washing performance and the textile hygiene should remain almost unchanged. Here in, the goal of the present study is to achieve an optimum EEP algorithm providing excellent textile hygiene results as well as cleaning performance in a domestic washing machine. In this regard, steam-pretreated cold wash approach with a combination of innovative algorithm solution in a relatively short washing cycle duration was implemented. For the parametric study, steam exposure time, washing load, total water consumption, main-washing time, and spinning rpm as the significant parameters affecting the textile hygiene and cleaning performance were investigated within a Design of Experiment study using Minitab 2021 statistical program. For the textile hygiene studies, specific loads containing the contaminated cotton carriers with Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa bacteria were washed. Then, the microbial removal performance of the designed programs was expressed as log reduction calculated as a difference of microbial count per ml of the liquids in which the cotton carriers before and after washing. For the cleaning performance studies, tests were carried out with various types of detergents and EMPA Standard Stain Strip. According to the results, the optimum EEP program provided an excellent hygiene performance of more than 2 log reduction of microorganism and a perfect Washing Efficiency Index (Iw) of 1.035, which is greater than the value specified by EU ecodesign regulation 2019/2023.Keywords: washing machine, energy efficient programs, hygiene, washing efficiency index, microorganism, escherichia coli, staphylococcus aureus, pseudomonas aeruginosa, laundry
Procedia PDF Downloads 1381674 The Effect of Some Macrofungi Extracts on Cytoplasmic Membrane of Multidrug Resistant Bacteria by Flow Cytometry
Authors: Yener Tekeli, Hayri Baba
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The natural active compounds found in medicinal plants are belong to various chemical structures including polyphenolic compounds, flavonoids, essential oils, and vitamins and some of these compounds have anticancer, antioxidant, and antimicrobial activity. However, these compounds have been little known about mechanisms to confer antibacterial drug resistance. In this study; some macrofungi extracts (Pholiota lucifera, Gnaoderma applanatum and Pleurotus ostreatus) were investigated for their abilities to enhance bacterial permeability by flow cytometry. This experiments exhibited enhancement of these extracts to disrupt the cytoplasmic membrane of living bacterial (Listeria innocua and Escherichia coli) cells. These experiments were designed to detect uptake of PI&SYT by enhancing with a ranged concentration of herb extracts.Keywords: antimicrobial activity, flow cytometry, macrofungi, multidrug resistant
Procedia PDF Downloads 4451673 Purification of Eicosapentaenoic Acid (EPA) and Docosahexaenoic Acid (DHA) from Fish Oil Using HPLC Method and Investigation of Their Antibacterial Effects on Some Pathogenic Bacteria
Authors: Yılmaz Uçar, Fatih Ozogul, Mustafa Durmuş, Yesim Ozogul, Ali Rıza Köşker, Esmeray Kuley Boğa, Deniz Ayas
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The aim of this study was to purified eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), that are essential oils from trout oil, using high-performance liquid chromatography (HPLC) method, bioconverted EPA and DHA into bioconverted EPA (bEPA), bioconverted DHA (bDHA) extracts by P. aeruginosa PR3. Moreover, in vitro antibacterial activity of bEPA and bDHA was investigated using disc diffusion methods and minimum inhibitory concentration (MIC). EPA and DHA concentration of 11.1% and 15.9% in trout oil increased in 58.64% and 40.33% after HPLC optimisation, respectively. In this study, EPA and DHA enriched products were obtained which are to be used as valuable supplements for food and pharmaceutical purposes. The bioconverted EPA and DHA exhibited antibacterial activities against two Gram-positive bacteria (Listeria monocytogenes ATCC 7677 and Staphylococcus aureus ATCC 29213) and six Gram-negative bacteria (Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC700603, Enterococcus faecalis ATCC 29212, Aeromonas hydrophila NCIMB 1135, and Salmonella Paratyphi A NCTC 13). Inhibition zones and MIC value of bEPA and bDHA against bacterial strains ranged from 7 to 12 mm and from 350 to 2350 μg/mL, respectively. Our results suggested that the crude extracts of bioconversion of EPA and DHA by P. aeruginosa PR3 can be considered as promising antimicrobials in improving food safety by controlling foodborne pathogens.Keywords: High-Performance Liquid Chromatography (HPLC), docosahexaenoic acid, DHA, eicosapentaenoic acid, EPA, minimum inhibitory concentration, MIC, Pseudomonas aeruginosa PR3
Procedia PDF Downloads 5011672 Prevalence of Foodborne Pathogens in Pig and Cattle Carcass Samples Collected from Korean Slaughterhouses
Authors: Kichan Lee, Kwang-Ho Choi, Mi-Hye Hwang, Young Min Son, Bang-Hun Hyun, Byeong Yeal Jung
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Recently, worldwide food safety authorities have been strengthening food hygiene in order to curb foodborne illness outbreaks. The hygiene status of Korean slaughterhouses has been monitored annually by Animal and Plant Quarantine Agency and provincial governments through foodborne pathogens investigation using slaughtered pig and cattle meats. This study presented the prevalence of food-borne pathogens from 2014 to 2016 in Korean slaughterhouses. Sampling, microbiological examinations, and analysis of results were performed in accordance with ‘Processing Standards and Ingredient Specifications for Livestock Products’. In total, swab samples from 337 pig carcasses (100 samples in 2014, 135 samples in 2015, 102 samples in 2016) and 319 cattle carcasses (100 samples in 2014, 119 samples in 2015, 100 samples in 2016) from twenty slaughterhouses were examined for Listeria monocytogenes, Campylobacter jejuni, Campylobacter coli, Salmonella spp., Staphylococcus aureus, Clostridium perfringens, Yersinia enterocolitica, Escherichia coli O157:H7 and non-O157 enterohemorrhagic E. coli (EHEC, serotypes O26, O45, O103, O104, O111, O121, O128 and O145) as foodborne pathogens. The samples were analyzed using cultural and PCR-based methods. Foodborne pathogens were isolated in 78 (23.1%) out of 337 pig samples. In 2014, S. aureus (n=17) was predominant, followed by Y. enterocolitica (n=7), C. perfringens (n=2) and L. monocytogenes (n=2). In 2015, C. coli (n=14) was the most prevalent, followed by L. monocytogenes (n=4), S. aureus (n=3), and C. perfringens (n=2). In 2016, S. aureus (n=16) was the most prevalent, followed by C. coli (n=13), L. monocytogenes (n=2) and C. perfringens (n=1). In case of cattle carcasses, foodborne bacteria were detected in 41 (12.9%) out of 319 samples. In 2014, S. aureus (n=16) was the most prevalent, followed by Y. enterocolitica (n=3), C. perfringens (n=3) and L. monocytogenes (n=2). In 2015, L. monocytogenes was isolated from 4 samples, S. aureus from three, C. perfringens, Y. enterocolitica and Salmonella spp. from one, respectively. In 2016, L. monocytogenes (n=6) was the most prevalent, followed by C. perfringens (n=3) C. jejuni (n=1), respectively. It was found that 10 carcass samples (4 cattle and 6 pigs) were contaminated with two bacterial pathogen tested. Interestingly, foodborne pathogens were more detected from pig carcasses than cattle carcasses. Although S. aureus was predominantly detected in this study, other foodborne pathogens were also isolated in slaughtered meats. Results of this study alerted the risk of foodborne pathogen infection for humans from slaughtered meats. Therefore, the authors insisted that it was important to enhance hygiene level of slaughterhouses according to Hazard Analysis and Critical Control Point.Keywords: carcass, cattle, foodborne, Korea, pathogen, pig
Procedia PDF Downloads 3441671 A Retrospective Cross Sectional Study of Blood Culture Results in a Tertiary Hospital, Ekiti, Nigeria
Authors: S. I. Nwadioha, M. S. Odimayo, J. A. Omotayo, A. Olu Taiwo, O. E. Olabiyi
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The current study was conducted to determine the epidemiology and antibiotic sensitivity pattern of bacteria isolated from blood of septicemic patients for improved antibiotic therapy. A three-year descriptive study has been carried out at Microbiology Laboratory, Ekiti State University Teaching Hospital, Ado Ekiti, from April 2012 to April 2015. Information compiled from patients’ records includes age, sex, isolated organisms and antibiotic susceptibility patterns. Three hundred and thirteen blood cultures were collected from neonatology and pediatrics wards, Out Patients’ Department (OPD) and from other adult patients. Forty-one cultures yielded mono microbial growth (no polymicrobial growth), giving an incidence of 13.1% positive blood culture (N=41/313). There were 58.4% Gram-negative bacilli and 41.6% Gram-positive cocci in the microbial growth. Bacteria isolated were Staphylococcus aureus 34%(14/41), Klebsiella species22% (9/41), Enterococci 17%(7/41), Proteus species12%(5/41), Escherichia coli 7%(3/41) and Streptococcal pneumoniae 7%(3/41). There was a (35%) higher occurrence of septicemia in neonates than in any other age groups in the hospital. Bacterial sensitivity to 13 antibiotic agents was determined by antibiotics disc diffusion using modified Kirby Bauer’s method. Gram-positive organisms showed a higher antibiotic sensitivity ranging from 14- 100% than the Gram-negative bacteria (11-80%). Staphylococcus aureus and Klebsiella species are the most prevalent organisms. The third generation Cephalosporins (Ceftriaxone) and Floroquinolone(Levofloxacin, Ofloxacin) have proved reliable for management of these blood infections.Keywords: blood cultures, septicemia, antibiogram, Nigeria
Procedia PDF Downloads 2351670 Molecular Profiling and Potential Bioactive Characteristics of Endophytic Fungi Isolated from Leptadenia Pyrotechnica
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
Procedia PDF Downloads 1411669 Antimicrobial Resistance Patterns of Campylobacter from Pig and Cattle Carcasses in Poland
Authors: Renata Szewczyk, Beata Lachtara, Kinga Wieczorek, Jacek Osek
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Campylobacter is recognized as the main cause of bacterial gastrointestinal infections in Europe. A main source of the pathogen is poultry and poultry meat; however, other animals like pigs and cattle can also be reservoirs of the bacteria. Human Campylobacter infections are often self-limiting but in some cases, macrolide and fluoroquinolones have to be used. The aim of this study was to determine antimicrobial resistance patterns (AMR) of Campylobacter isolated from pig and cattle carcasses. Between July 2009 and December 2015, 735 swabs from pig (n = 457) and cattle (n = 278) carcasses were collected at Polish slaughterhouses. All samples were tested for the presence of Campylobacter by ISO 10272-1 and confirmed to species level using PCR. The antimicrobial susceptibility of Campylobacter isolates was determined by a microbroth dilution method with six antimicrobials: gentamicin (GEN), streptomycin (STR), erythromycin (ERY), nalidixic acid (NAL), ciprofloxacin (CIP) and tetracycline (TET). It was found that 167 of 735 samples (22.7%) were contaminated with Campylobacter. The vast majority of them were of pig origin (134; 80.2%), whereas for cattle carcasses Campylobacter was less prevalent (33; 19.8%). Among positive samples C. coli was predominant species (123; 73.7%) and it was isolated mainly from pig carcasses. The remaining isolates were identified as C. jejuni (44; 26.3%). Antimicrobial susceptibility indicated that 22 out of 167 Campylobacter (13.2%) were sensitive to all antimicrobials used. Fourteen of them were C. jejuni (63.6%; pig, n = 6; cattle, n = 8) and 8 was C. coli (36.4%; pig, n = 4; cattle, n = 4). Most of the Campylobacter isolates (145; 86.8%) were resistant to one or more antimicrobials (C. coli, n = 115; C. jejuni, n = 30). Comparing the AMR for Campylobacter species it was found that the most common pattern for C. jejuni was CIP-NAL-TET (9; 30.0%), whereas CIP-NAL-STR-TET was predominant among C. coli (47; 40.9%). Multiresistance, defined as resistance to three or more classes of antimicrobials, was found in 57 C. coli strains, mostly obtained from pig (52 isolates). On the other hand, only one C. jejuni strain, isolated from cattle, showed multiresistance with pattern CIP-NAL-STR-TET. Moreover, CIP-NAL-STR-TET was characteristic for most of multiresistant C. coli isolates (47; 82.5%). For the remaining C. coli the resistance patterns were CIP-ERY-NAL-TET (7 strains; 12.3%) and for one strain of each patterns: ERY-STR-TET, CIP-STR-TET, CIP-NAL-GEN-STR-TET. According to the present findings resistance to erythromycin was observed only in 11 C. coli (pig, n = 10; cattle, n = 1). In conclusion, the results of this study showed that pig carcasses may be a serious public health concern because of contamination with C. coli that might features multiresistance to antimicrobials.Keywords: antimicrobial resistance, Campylobacter, carcasses, multi resistance
Procedia PDF Downloads 3341668 Pathogenic Escherichia Coli Strains and Their Antibiotic Susceptibility Profiles in Cases of Child Diarrhea at Addis Ababa University, College of Health Sciences, Tikur Anbessa Specialized Hospital, Addis Ababa, Ethiopia
Authors: Benyam Zenebe, Tesfaye Sisay, Gurja Belay, Workabeba Abebe
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Background: The prevalence and antibiogram of pathogenic E. coli strains, which cause diarrhea vary from region to region, and even within countries in the same geographical area. In Ethiopia, diagnostic approaches to E. coli induced diarrhea in children less than five years of age are not standardized. The aim of this study was to determine the involvement of pathogenic E. coli strains in child diarrhea and determine the antibiograms of the isolates in children less than 5 years of age with diarrhea at Addis Ababa University College of Health Sciences TikurAnbessa Specialized Hospital, Addis Ababa, Ethiopia. Methods: A purposive study that included 98 diarrheic children less than five years of age was conducted at Addis Ababa University College of Health Sciences, TikurAnbessa Specialized Hospital, Addis Ababa, Ethiopia to detect pathogenic E. coli biotypes. Stool culture was used to identify presumptive E. coliisolates. Presumptive isolates were confirmed by biochemical tests, and antimicrobial susceptibility tests were performed on confirmed E. coli isolates by the disk diffusion method. DNA was extracted from confirmed isolates by a heating method and subjected to Polymerase Chain Reaction or the presence of virulence genes. Amplified PCR products were analyzed by agarose gel electrophoresis. Data were collected on child demographics and clinical conditions using administered questionnaires. The prevalence of E. coli strains from the total diarrheic children, and the prevalence of pathogenic strains from total E. coli isolates along with their susceptibility profiles; the distribution of pathogenic E.coli biotypes among different age groups and between the sexes were determined by using descriptive statistics. Result: Out of 98 stool specimens collected from diarrheic children less than 5 years of age, 75 presumptive E. coli isolates were identified by culture; further confirmation by biochemical tests showed that only 56 of the isolates were E. coli; 29 of the isolates were found in male children and 27 of them in female children. Out of the 58 isolates of E. coli, 25 pathotypes belonging to different classes of pathogenic strains: STEC, EPEC, EHEC, EAEC were detected by using the PCR technique. Pathogenic E. coli exhibited high rates of antibiotic resistance to many of the antibiotics tested. Moreover, they exhibited multiple drug resistance. Conclusion: This study found that the isolation rate of E. coli and the involvement of antibiotic-resistant pathogenic E. coli in diarrheic children is prominent, and hence focus should be given on the diagnosis and antimicrobial sensitivity testing of pathogenic E. coli at Addis Ababa University College of Health Sciences TikurAnbessa Specialized Hospital. Among antibiotics tested, Cefotitan could be a drug of choice to treat E. coli.Keywords: antibiotic susceptibility profile, children, diarrhea, E. coli, pathogenic
Procedia PDF Downloads 2341667 Safety Assessment of Traditional Ready-to-Eat Meat Products Vended at Retail Outlets in Kebbi and Sokoto States, Nigeria
Authors: M. I. Ribah, M. Jibir, Y. A. Bashar, S. S. Manga
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Food safety is a significant and growing public health problem in the world and Nigeria as a developing country, since food-borne diseases are important contributors to the huge burden of sickness and death of humans. In Nigeria, traditional ready-to-eat meat products (RTE-MPs) like balangu, tsire, guru and dried meat products like kilishi, dambun nama, banda, were reported to be highly appreciated because of their eating qualities. The consumption of these products was considered as safe due to the treatments that are usually involved during their production process. However, during processing and handling, the products could be contaminated by pathogens that could cause food poisoning. Therefore, a hazard identification for pathogenic bacteria on some traditional RTE-MPs was conducted in Kebbi and Sokoto States, Nigeria. A total of 116 RTE-MPs (balangu-38, kilishi-39 and tsire-39) samples were obtained from retail outlets and analyzed using standard cultural microbiological procedures in general and selective enrichment media to isolate the target pathogens. A six-fold serial dilution was prepared and using the pour plating method, colonies were counted. Serial dilutions were selected based on the prepared pre-labeled Petri dishes for each sample. A volume of 10-12 ml of molten Nutrient agar cooled to 42-45°C was poured into each Petri dish and 1 ml each from dilutions of 102, 104 and 106 for every sample was respectively poured on a pre-labeled Petri plate after which colonies were counted. The isolated pathogens were identified and confirmed after series of biochemical tests. Frequencies and percentages were used to describe the presence of pathogens. The General Linear Model was used to analyze data on pathogen presence according to RTE-MPs and means were separated using the Tukey test at 0.05 confidence level. Of the 116 RTE-MPs samples collected, 35 (30.17%) samples were found to be contaminated with some tested pathogens. Prevalence results showed that Escherichia coli, salmonella and Staphylococcus aureus were present in the samples. Mean total bacterial count was 23.82×106 cfu/g. The frequency of individual pathogens isolated was; Staphylococcus aureus 18 (15.51%), Escherichia coli 12 (10.34%) and Salmonella 5 (4.31%). Also, among the RTE-MPs tested, the total bacterial counts were found to differ significantly (P < 0.05), with 1.81, 2.41 and 2.9×104 cfu/g for tsire, kilishi, and balangu, respectively. The study concluded that the presence of pathogenic bacteria in balangu could pose grave health risks to consumers, and hence, recommended good manufacturing practices in the production of balangu to improve the products’ safety.Keywords: ready-to-eat meat products, retail outlets, public health, safety assessment
Procedia PDF Downloads 1341666 Characterization of Shiga Toxin Escherichia coli Recovered from a Beef Processing Facility within Southern Ontario and Comparative Performance of Molecular Diagnostic Platforms
Authors: Jessica C. Bannon, Cleso M. Jordao Jr., Mohammad Melebari, Carlos Leon-Velarde, Roger Johnson, Keith Warriner
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There has been an increased incidence of non-O157 Shiga Toxin Escherichia coli (STEC) with six serotypes (Top 6) being implicated in causing haemolytic uremic syndrome (HUS). Beef has been suggested to be a significant vehicle for non-O157 STEC although conclusive evidence has yet to be obtained. The following aimed to determine the prevalence of the Top 6 non-O157 STEC in beef processing using three different diagnostic platforms then characterize the recovered isolates. Hide, carcass and environmental swab samples (n = 60) were collected from a beef processing facility over a 12 month period. Enriched samples were screened using Biocontrol GDS, BAX or PALLgene molecular diagnostic tests. Presumptive non-O157 STEC positive samples were confirmed using conventional PCR and serology. STEC was detected by GDS (55% positive), BAX (85% positive), and PALLgene (93%). However, during confirmation testing only 8 of the 60 samples (13%) were found to harbour STEC. Interestingly, the presence of virulence factors in the recovered isolates was unstable and readily lost during subsequent sub-culturing. There is a low prevalence of Top 6 non-O157 STEC associated with beef although other serotypes are encountered. Yet, the instability of the virulence factors in recovered strains would question their clinical relevance.Keywords: beef, food microbiology, shiga toxin, STEC
Procedia PDF Downloads 4631665 Biosynthesis of Silver Nanoparticles Using Zataria multiflora Extract, and Study of Antibacterial Effects on UTI Bacteria (MDR)
Authors: Mohammad Hossein Pazandeh, Monir Doudi, Sona Rostampour Yasouri
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Irregular consumption of current antibiotic makes increases of antibiotic resistance between urin pathogens on all worlds. This study selected based on this great community problem. The aim of this study was the biosynthesis of silver nanoparticles from Zataria multiflora extract and then to investigate its antibacterial effect on gram-negative bacilli common in Urinary Tract Infections (UTI) and MDR. The plant used in the present research was Zataria multiflora whose extract was prepared through Soxhlet extraction method. Green synthesis condition of silver nanoparticles was investigated in terms of three parameters including the extract amount, concentration of silver nitrate salt, and temperature. The seizes of nanoparticles were determined by Zetasizer. In order to identify synthesized silver nanoparticles Transmission Electron Microscopy (TEM) and X-ray Diffraction (XRD) methods were used. For evaluating the antibacterial effects of nanoparticles synthesized through biological method different concentrations of silver nanoparticles were studied on 140 cases of Muliple Drug Resistance (MDR) bacteria strains Escherichia coli, Klebsiella pneumoniae, Enterobacter aerogenes, Proteus vulgaris,Citrobacter freundii, Acinetobacter bumanii and Pseudomonas aeruginosa, (each genus of bacteria, 20 samples), which all were MDR and cause urinary tract infections , for identification of bacteria were used of Polymerase Chain Reaction (PCR) test and laboratory methods (Agar well diffusion and Microdilution methods) to assess their sensitivity to Nanoparticles. The data were analyzed using SPSS software by nonparametric Kruskal-Wallis and Mann-Whitney tests. Significant results were found about the effects of silver nitrate concentration, different amounts of Zataria multiflora extract, and temperature on nanoparticles; that is, by increasing the concentration of silver nitrate, extract amount, and temperature, the sizes of synthesized nanoparticles declined. However, the effect of above mentioned factors on particles diffusion index was not significant. Based on the TEM results, particles were mainly spherical shape with a diameter range of 25 to 50 nm. The results of XRD Analysis indicated the formation of Nanostructures and Nanocrystals of silver.. The obtained results of antibacterial effects of different concentrations of silver nanoparticles on according to agar well diffusion and microdilution method, biologically synthesized nanoparticles showed 1000 mg /ml highest and lowest mean inhibition zone diameter in E.coli , Acinetobacter bumanii 23 and 15mm, respectively. MIC was observed for all of bacteria 125mg/ml and for Acinetobacter bumanii 250mg/ml.Comparing the growth inhibitory effect of chemically synthesized Nanoparticles and biologically synthesized Nanoparticles showed that in the chemical method the highest growth inhibition belonged to the concentration of 62.5 mg /ml. The inhibitory effect on the growth all of bacteria causes of urine infection and MDR was observed and by increasing silver ion concentration in Nanoparticles, antibacterial activity increased. Generally, the biological synthesis can be considered an efficient way not only in making Nanoparticles but also for having anti-bacterial properties. It is more biocompatible and may be possess less toxicity than the Nanoparticles synthesized chemically.Keywords: biosynthesis, MDR bacteria, silver nanoparticles, UTI
Procedia PDF Downloads 541664 An Evaluative Microbiological Risk Assessment of Drinking Water Supply in the Carpathian Region: Identification of Occurrent Hazardous Bacteria with Quantitative Microbial Risk Assessment Method
Authors: Anikó Kaluzsa
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The article's author aims to introduce and analyze those microbiological safety hazards which indicate the presence of secondary contamination in the water supply system. Since drinking water belongs to primary foods and is the basic condition of life, special attention should be paid on its quality. There are such indicators among the microbiological features can be found in water, which are clear evidence of the presence of water contamination, and based on this there is no need to perform other diagnostics, because they prove properly the contamination of the given water supply section. Laboratory analysis can help - both technologically and temporally – to identify contamination, but it does matter how long takes the removal and if the disinfection process takes place in time. The identification of the factors that often occur in the same places or the chance of their occurrence is greater than the average, facilitates our work. The pathogen microbiological risk assessment by the help of several features determines the most likely occurring microbiological features in the Carpathian basin. From among all the microbiological indicators, that are recommended targets for routine inspection by the World Health Organization, there is a paramount importance of the appearance of Escherichia coli in the water network, as its presence indicates the potential ubietiy of enteric pathogens or other contaminants in the water network. In addition, the author presents the steps of microbiological risk assessment analyzing those pathogenic micro-organisms registered to be the most critical.Keywords: drinking water, E. coli, microbiological indicators, risk assessment, water safety plan
Procedia PDF Downloads 3341663 In vitro Antioxidant, Anticancer Properties and Probiotic Characteristics of Selected Lactic Acid Bacteria Strains
Authors: M. G. Shehata, S. A. El Sohaimy, Marwa M. Abu-Serie, Nourhan M. Abd El-Aziz
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Probiotic strains can potentially be used as bio-preservatives and functional food supplement. Eight lactic acid bacteria strains (LAB) Lactobacillus brevis NRRL B-4527; Streptococcus thermophilus BLM 58; Pediococcusacidilactici ATCC 8042; Lactobacillus rhamnosus CCUG 1452; Lactobacillus curvatus ATCC 51436; Lactococcuslactis sub sp. lactisDSM 20481; Lactobacillus plantarum DMSZ 20079 and Lactobacillus plantarumTF103 were selected to screen the antioxidant, anticancer potential and probiotic properties. LAB strains exhibited good probiotic, antioxidant properties and showed antagonistic activity against food-borne pathogenic (Bacillus subtilis DB 100 host; Candida albicans ATCCMYA-2876; Clostridium botulinum ATCC 3584; Escherichia coli BA 12296; Klebsiellapneumoniae ATCC12296; Salmonella senftenberg ATCC 8400 and Staphylococcus aureus NCTC 10788). Further, in vitro probiotic properties of eight strains displayed excellent acid tolerance, bile tolerance, simulated gastrointestinal juice tolerance, in vitro adhesion ability for HT-29 cell line. The antioxidant effect of intracellular and cell-free extract of lactic acid bacteria strains was evaluated by various antioxidant assays, namely, resistance to hydrogen peroxide, DPPH radical scavenging, ABTS radical scavenging, and hydroxyl radical scavenging (HRS). The results showed that intracellular and cell-free supernatant of S. Thermophilus BLM 58, L. lactissubsp.lactis DSM 20481, P. acidilactici ATCC 8042, L. brevis NRRL B-4527 strains possess excellent antioxidant capacity. The intracellular of S. Thermophilus BLM 58 and P. acidilactici ATCC 8042 also showed excellent anticancer activity against Caco-2, MCF-7, HepG-2, and PC-3. Antioxidative property of selected lactic acid bacteria strains would be useful in the functional food manufacturing industry. They could beneficially affect the consumer by providing dietary source of antioxidants.Keywords: anticancer activity, antioxidant activity, functional food, lactic acid bacteria, probiotic
Procedia PDF Downloads 2241662 Antimicrobial Activity of Igusa and the Application to Foam Materials for Food Industry
Authors: I. Nanako, Mariko Era, Hiroshi Morita
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Objectives: Japanese uses TATAMI rather than flooring at home. Igusa ( Juncus effuses var. decipiens ), which is commonly known in the forms of TATAMI. Juncus spp. grow at a relatively high humidity area (Japan, China and Southeast Asia ). Yatsushiro region in the southern part of Kumamoto prefecture is major produing area of Igusa. Igusa found to have honeycomb structure and was also shown to have the ability to control humidity. And Igusa has been used as a medicinal herb for diuretic and antiphlogistic agent. In previous study, we investigated antimicrobial effects of Igusa, and showed high antimicrobial activity against food poisoning bacteria. Therefore, the food trays blended Igusa can be kept clean by antimicrobial activity of Igusa. We focus on ‘Igusa foam materials’. In this study, we investigated the antibacterial and antifungal activity of Igusa, and new application to foam materials for food industry. Materials and method: We used Igusa foam materials (3 × 3 × 3 cm) as a sample. We set about fifteen types of samples combined with a commercial antibacterial agent A, a commercial antibacterial agent B, potassium laurate (C12K) and a commercial antifungal agent C, a commercial antifungal agent D and a commercial antifungal agent E. We selected four bacteria strains (Escherichia coli NBRC 3972, Staphylococus aureus NBRC 12732, Salmonella typhimurium NBRC 13245, Bacillus subtilis NBRC 3335 ) and three fungus strains (Penicillium pinophilum NBRC 6345, Cladosporium cladosporioides NBRC 30314, Aspergillus oryzae NBRC 5238 ). The fungus was cultured at 30 °C on Igusa foam materials after inoculation of the fungus for fourteen days. The bacteria was cultured at 30 °C on Igusa foam materials after inoculation of the bacteria for three days. And the Igusa foam materials were washed with 10 mL normal saline after three days. The normal saline washed Igusa foam materials plated the NA medium. After, It was cultured at 30 °C and used colony counting method. Result and Conclusion: The fifteen types of sample of Igusa foam materials had antifungal activity against C. cladosporioides, A. oryzae and P. pinophilum for fourteen days. The four types of sample contained potassium laurate and antibacterial agent A, sample contained antibacterial agent B and antifungal agent D, sample contained A and antifungal agent E, sample contained B and E had antibacterial activity against B. subtilis. The three types of sample contained potassium laurate and A, sample contained B and D, sample contained A and E had antibacterial activity against S. typhimurium. The five types of sample contained potassium laurate and A, sample contained B and D, sample contained A and E, sample contained B and E, sample contained B and antifungal agent C had antibacterial activity against E. coli and S. aureus. These results indicate that Igusa of Igusa foam materials had high antifungal activity. In addition, Igusa foam materials combined with a commercial antibacterial agent had antibacterial activity. In the future, we consider that use of Igusa foam materials may be spread from food industry.Keywords: antibacterial, antifungal, foam materials, Igusa
Procedia PDF Downloads 2451661 Synthesis, Characterization and Anti-Microbial Study of Urethanized Poly Vinyl Alcohol Metal Complexes
Authors: Maha A. Younus, Dhefaf H. Badri, Maha A. Al Abayaji, Taha M. Salih
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Polymer metal complexes of poly vinyl alcohol and Cu (II), Ni (II), Mn (II) and Co (III) were prepared from the reaction of PVA with three different percentages of urea. The compound was characterized by fourier transform infrared spectrometry (FTIR) analysis and differential scanning calorimetric (DSC) Analysis. It has been established that the polymer and its metal complexes showed good activities against nine pathogenic bacteria (Escherichia coli, Klebsiellapneumonae, Staphylococcusaureus, Staphylococcus Albus, Salmonella Typhoid, Pseudomonas Aeruginosa, Shigella Dysentery, Proteus Morgani, Brucella Militensis). The polymer metal complexes show activity higher than that of the free polymer. The increasing activities were in the order (polymer < pol-Mn< pol-Co < pol-Ni ˂ pol-Cu). The ability of these compounds to show antimicrobial properties suggests that they can be further evaluated for medicinal and/or environmental applications.Keywords: antimicrobial activity, PVA, polymer-metal complex, urea
Procedia PDF Downloads 3381660 Polyphosphate Kinase 1 Active Site Characterization for the Identification of Novel Antimicrobial Targets
Authors: Sanaa Bardaweel
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Inorganic polyphosphate (poly P) is present in all living forms tested to date, from each of the three kingdoms of life. Studied mainly in prokaryotes, poly P and its associated enzymes are vital in diverse basic metabolism, in at least some structural functions and, notably, in stress responses. These plentiful and unrelated roles for poly P are probably the consequence of its presence in life-forms early in evolution. The genomes of many bacterial species, including pathogens, encode a homologue of a major poly P synthetic enzyme, poly P kinase 1 (PPK1). Genetic deletion of ppk1 results in reduced poly P levels and loss of pathogens virulence towards protozoa and animals. Thus far, no PPK1 homologue has been identified in higher-order eukaryotes and, therefore, PPK1 represents a novel target for chemotherapy. The idea of the current study is to purify the PPK1 from Escherichia coli to homogeneity in order to study the effect of active site point mutations on PPK1 catalysis via the application of site-directed mutagenesis strategy. The knowledge obtained about the active site of PPK1 will be utilized to characterize the catalytic and kinetic mechanism of PPK1 with model substrates. Comprehensive understanding of the enzyme kinetic mechanism and catalysis will be used to design and screen a library of synthetic compounds for potential discovery of selective PPK1-inhibitors.Keywords: antimicobial, Escherichia coli, inorganic polyphosphate, PPK1-inhibitors
Procedia PDF Downloads 2791659 Effect of Lime Stabilization on E. coli Destruction and Heavy Metal Bioavailability in Sewage Sludge for Agricultural Utilization
Authors: G. Petruzzelli, F. Pedron, M. Grifoni, A. Pera, I. Rosellini, B. Pezzarossa
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The addition of lime as Ca(OH)2 to sewage sludge to destroy pathogens (Escherichia coli), was evaluated also in relation to heavy metal bioavailability. The obtained results show that the use of calcium hydroxide at the dose of 3% effectively destroyed pathogens ensuring the stability at high pH values over long period and the duration of the sewage sludge stabilization. In general, lime addition decreased the total extractability of heavy metals indicating a reduced bioavailability of these elements. This is particularly important for a safe utilization in agricultural soils to reduce the possible transfer of heavy metals to the food chain.Keywords: biological sludge, Ca(OH)2, copper, pathogens, sanitation, zinc
Procedia PDF Downloads 4261658 A contribution to Phytochemical and Biological Studies of Ailanthus Alitssima Swingle Cultivated in Egypt
Authors: Ahmed Samy Elnoby
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Ailanthus altissima native to Asia which belongs to the family Simaroubaceae was subjected to phytochemical screening and biological investigations. Phytochemical screening revealed the presence of carbohydrates, tannins, sterols, flavonoids and traces of saponins. In addition, quantitative determination of phenolics and flavonoid content were performed. The antimicrobial activity of methanolic extract of the leaves was determined against gram-positive, gram-negative bacteria in addition to fungi using a modified Kirby-Bauer disc diffusion method that was compared with standard discs ampicillin which acts as an antibacterial agent and amphotericin B which acts as an antifungal agent. A high potency was observed against gram-positive bacteria mainly staphylococcus aureus, gram-negative bacteria mainly Escherichia coli and showed no potency against fungi mainly Aspergillus flavus and candida albicans. On the other hand, the antioxidant activity of the extract was determined by 1, 1-diphenyl-2- diphenyl-2-picryl-hydrazil (DPPH). A very low potency was shown by using DPPH for the antioxidant effect so IC50 = 0 ug/ml, IC90 =0 ug /ml and remark gave 47.2 % at 100 ug/ml which is very weak. Cytotoxic activity was determined by using MTT assay (3-4, 5-Dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium Bromide) against MCF7 (Human Caucasian breast adenocarcinoma) cell line. A moderate potency was shown by using MCF7 cell line for cytotoxic effect so LC50= 90.2 ug/ml, LC90=139.9 ug/ml and the remark gave 55.2% at 100 ug/ml which is of moderate activity so, Ailanthus altissima can be considered to be a promising antimicrobial agent from natural origin.Keywords: Ailanthus altissima, TLC, HPLC, anti-microbial activity, antifungal activity, antioxidant, cytotoxic activity
Procedia PDF Downloads 1751657 Antimicrobial Properties of SEBS Compounds with Copper Microparticles
Authors: Vanda Ferreira Ribeiro, Daiane Tomacheski, Douglas Naue Simões, Michele Pitto, Ruth Marlene Campomanes Santana
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Indoor environments, such as car cabins and public transportation vehicles are places where users are subject to air quality. Microorganisms (bacteria, fungi, yeasts) enter these environments through windows, ventilation systems and may use the organic particles present as a growth substrate. In addition, atmospheric pollutants can act as potential carbon and nitrogen sources for some microorganisms. Compounds base SEBS copolymers, poly(styrene-b-(ethylene-co-butylene)-b-styrene, are a class of thermoplastic elastomers (TPEs), fully recyclable and largely used in automotive parts. Metals, such as cooper and silver, have biocidal activities and the production of the SEBS compounds by melting blending with these agents can be a good option for producing compounds for use in plastic parts of ventilation systems and automotive air-conditioning, in order to minimize the problems caused by growth of pathogenic microorganisms. In this sense, the aim of this work was to evaluate the effect of copper microparticles as antimicrobial agent in compositions based on SEBS/PP/oil/calcite. Copper microparticles were used in weight proportion of 0%, 1%, 2% and 4%. The compounds were prepared using a co-rotating double screw extruder (L/D ratio of 40/1 and 16 mm screw diameter). The processing parameters were 300 rpm of screw rotation rate, with a temperature profile between 150 to 190°C. SEBS based TPE compounds were injection molded. The compounds emission were characterized by gravimetric fogging test. Compounds were characterized by physical (density and staining by contact), mechanical (hardness and tension properties) and rheological properties (melt volume rate – MVR). Antibacterial properties were evaluated against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) strains. To avaluate the abilities toward the fungi have been chosen Aspergillus niger (A. niger), Candida albicans (C. albicans), Cladosporium cladosporioides (C. cladosporioides) and Penicillium chrysogenum (P. chrysogenum). The results of biological tests showed a reduction on bacteria in up to 88% in E.coli and up to 93% in S. aureus. The tests with fungi showed no conclusive results because the sample without copper also demonstrated inhibition of the development of these microorganisms. The copper addition did not cause significant variations in mechanical properties, in the MVR and the emission behavior of the compounds. The density increases with the increment of copper in compounds.Keywords: air conditioner, antimicrobial, cooper, SEBS
Procedia PDF Downloads 2831656 Antibacterial Activity of Calendula officinalis Extract Loaded Chitosan Nanoparticles
Authors: Sanjay Singh, Swati Jaiswal, Prashant Mishra
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Nanoparticle based formulations of drug delivery systems have shown their potential in improving the performance of existing drugs and have opened avenues for new therapies. Calendula extract is a low cost, wide spectrum bioactive material that has been used for a long term therapy of various infections. Aim: The aim of this study was to develop Calendula officinalis extract based nanoformulations and to study the antibacterial activity of either Calendula extract loaded chitosan nanoparticles or Calendula extract coated silver nanoparticles for increased bioavailability and their long term effect. Methods: Chitosan nanoparticles were prepared by the process of ionotropic gelation, based on interaction between the negative groups of tri polyphosphate (TPP) and positively charged amino groups of chitosan. The size of the Calendula extract-loaded chitosan particles was determined using dynamic light scattering and scanning electron microscopy. Antibacterial activities of these formulations were determined based on minimum inhibitory concentration and time kill studies. In addition, silver nanoparticles were also synthesized in the presence of Calendula extract and characterized by UV visible spectrum, DLS and XRD. Experiments were conducted on 96-plates against two Gram-positive bacteria; Staphylococcus aureus and Bacillus subtilis two Gram-negative bacteria; Escherichia coli and Pseudomonas aeruginosa. Results: Results demonstrated time dependent antibacterial activity against different microbes studied. Both Calendula extract and Calendula extract loaded chitosan nanoparticles have shown good antimicrobial activity against both Gram positive and Gram negative bacteria. Conclusion: Calendula extract loaded chitosan nanoparticles and calendula extract coated silver nanoparticles are potential antibacterial for their long term antibacterial effects.Keywords: antibacterial, Calendula extract, chitosan nanoparticles, silver nanoparticles
Procedia PDF Downloads 3461655 Exploratory Tests of Crude Bacteriocins from Autochthonous Lactic Acid Bacteria against Food-Borne Pathogens and Spoilage Bacteria
Authors: M. Naimi, M. B. Khaled
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The aim of the present work was to test in vitro inhibition of food pathogens and spoilage bacteria by crude bacteriocins from autochthonous lactic acid bacteria. Thirty autochthonous lactic acid bacteria isolated previously, belonging to the genera: Lactobacillus, Carnobacterium, Lactococcus, Vagococcus, Streptococcus, and Pediococcus, have been screened by an agar spot test and a well diffusion assay against Gram-positive and Gram-negative harmful bacteria: Bacillus cereus, Bacillus subtilis ATCC 6633, Escherichia coli ATCC 8739, Salmonella typhimurium ATCC 14028, Staphylococcus aureus ATCC 6538, and Pseudomonas aeruginosa under conditions means to reduce lactic acid and hydrogen peroxide effect to select bacteria with high bacteriocinogenic potential. Furthermore, crude bacteriocins semiquantification and heat sensitivity to different temperatures (80, 95, 110°C, and 121°C) were performed. Another exploratory test concerning the response of St. aureus ATCC 6538 to the presence of crude bacteriocins was realized. It has been observed by the agar spot test that fifteen candidates were active toward Gram-positive targets strains. The secondary screening demonstrated an antagonistic activity oriented only against St. aureus ATCC 6538, leading to the selection of five isolates: Lm14, Lm21, Lm23, Lm24, and Lm25 with a larger inhibition zone compared to the others. The ANOVA statistical analysis reveals a small variation of repeatability: Lm21: 0.56%, Lm23: 0%, Lm25: 1.67%, Lm14: 1.88%, Lm24: 2.14%. Conversely, slight variation was reported in terms of inhibition diameters: 9.58± 0.40, 9.83± 0.46, and 10.16± 0.24 8.5 ± 0.40 10 mm for, Lm21, Lm23, Lm25, Lm14and Lm24, indicating that the observed potential showed a heterogeneous distribution (BMS = 0.383, WMS = 0.117). The repeatability coefficient calculated displayed 7.35%. As for the bacteriocins semiquantification, the five samples exhibited production amounts about 4.16 for Lm21, Lm23, Lm25 and 2.08 AU/ml for Lm14, Lm24. Concerning the sensitivity the crude bacteriocins were fully insensitive to heat inactivation, until 121°C, they preserved the same inhibition diameter. As to, kinetic of growth , the µmax showed reductions in pathogens load for Lm21, Lm23, Lm25, Lm14, Lm24 of about 42.92%, 84.12%, 88.55%, 54.95%, 29.97% in the second trails. Inversely, this pathogen growth after five hours displayed differences of 79.45%, 12.64%, 11.82%, 87.88%, 85.66% in the second trails, compared to the control. This study showed potential inhibition to the growth of this food pathogen, suggesting the possibility to improve the hygienic food quality.Keywords: exploratory test, lactic acid bacteria, crude bacteriocins, spoilage, pathogens
Procedia PDF Downloads 2131654 An Investigation of How Salad Rocket May Provide Its Own Defence Against Spoilage Bacteria
Authors: Huda Aldossari
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Members of the Brassicaceae family, such as rocket species, have high concentrations of glucosinolates (GLSs). GSLs and isothiocyanates (ITCs), the product of GLSs hydrolysis, are the most influential compounds that affect flavour in rocket species. Aside from their contribution to the flavour, GSLs and ITCs are of particular interest due to their potential ability to inhibit the growth of human pathogenic bacteria such as E. coli O157. Quantitative and qualitative analysis of glucosinolate compounds in rocket extracts was obtained by Liquid Chromatography-Mass Spectrometry (LC–MS).Each individual component of non-volatile GLSs and ITCs was isolated by High-Performance Liquid Chromatography (HPLC) fractionation. The identity and purity of each fraction were confirmed using Ultra High-Performance Liquid Chromatography (UPLC). The separation of glucosinolates in the complex rocket extractions was performed by optimizing a HPLC fractionation method through changing the mobile phase composition, solvent gradient, and the flow rate. As a result, six glucosinolates compounds (Glucosativin, 4-Methoxyglucobrassicin, Glucotropaeolin GTP, Glucoiberin GIB, Diglucothiobenin, and Sinigrin) have been isolated, identified and quantified in the complex samples. This step aims to evaluate the antibacterial activity of glucosinolates and their enzymatic hydrolysis against bacterial growth of E.coli k12. Therefore, fractions from this study will be used to determine the most active compounds by investigating the efficacy of each component of GLSs and ITCs at inhibiting bacterial growth.Keywords: rocket, glucosinolates, E.coli k12., HPLC fractionatio
Procedia PDF Downloads 981653 Antimicrobial Properties of Copper in Gram-Negative and Gram-Positive Bacteria
Authors: Travis J. Meyer, Jasodra Ramlall, Phyo Thu, Nidhi Gadura
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For centuries humans have used the antimicrobial properties of copper to their advantage. Yet, after all these years the underlying mechanisms of copper mediated cell death in various microbes remain unclear. We had explored the hypothesis that copper mediated increased levels of lipid peroxidation in the membrane fatty acids is responsible for increased killing inEscherichia coli. In this study we show that in both gram positive (Staphylococcus aureus) and gram negative (Pseudomonas aeruginosa) bacteria there is a strong correlation between copper mediated cell death and increased levels of lipid peroxidation. Interestingly, the non-spore forming gram positive bacteria as well as gram negative bacteria show similar patterns of cell death, increased levels of lipid peroxidation, as well as genomic DNA degradation, however there is some difference inloss in membrane integrity upon exposure to copper alloy surface.Keywords: antimicrobial, copper, gram positive, gram negative
Procedia PDF Downloads 4831652 Identification of Phenolic Compounds with Antibacterial Activity in Raisin Extract
Authors: Yousef M. Abouzeed A. Elfahem, F. Zgheel, M. A. Saad, Mohamed O. Ahmed
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The bioactive properties of phytochemicals indicate their potential as natural drug products to prevent and treat human disease; in particular, compounds with antioxidant and antimicrobial activities may represent a novel class of safe and effective drugs. Following desiccation, grapes (Vitis vinifera) become more resistant to microbial-based degradation, suggesting that raisins may be a source of antimicrobial compounds. To investigate this hypothesis, total phenolic extracts were obtained from common raisins, local market-sourced. The acetone extract was tested for antibacterial activity against four prevalent bacterial pathogens (Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella spp. and Escherichia coli). Antibiotic sensitivity and the Minimum Inhibitory Concentration (MIC) were determined for each bacterium. High performance liquid chromatography was used to identify compounds in the total phenolic extract. The raisin phenolic extract inhibited growth of all the tested bacteria; the greatest inhibitive effect (normalized to cefotaxime sodium control antibiotic) occurred against P. aeruginosa, followed by S. aureus > Salmonella spp.= E. coli. The phenolic extracts contained the bioactive compounds catechin, quercetin, and rutin. Thus, phytochemicals in raisin extract have antibacterial properties; this plant-based extract, or its bioactive constituents, may represent a promising natural preservative or antimicrobial agent for the food industry or anti-infective drug.Keywords: Vitis vinifera raisin, extraction, phenolic compounds, antibacterial activity
Procedia PDF Downloads 6061651 Microbial Quality of Traditional Qatari Foods Sold by Women Street Vendors in Doha, Qatar
Authors: Tahra El-Obeid, Reham Mousa, Amal Alzahiri
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During the past few years the traditional market of Qatar has become an attraction to many customers who eat from the numerous women street vendors selling Qatari traditional dishes. To gain an understanding on the safety of these street vended foods, we designed the study to test microbiological quality of 14 different Qatari foods sold in Souk Wagif, the main traditional market in Qatar. This study was conducted to mainly identify presence or absence of microbial pathogens. A total of 56 samples were purchased from 10 different street vendors and the samples were collected randomly on different days. The samples were tested for microbial contaminants at Central Food Laboratories, Doha, Qatar. The qualitative study was conducted using Real Time-PCR to screen for; Salmonella spp., Listeria monocytogenes, Escherichia coli and E. coli 0157:H7. Out of the 56 samples, only two samples “Biryani” and “Khabess” contained E. coli. However, both samples tested negative for E. coli O157:H7. The microbial contamination of the Qatari traditional street vended foods was 3%. This result may be attributed to the food safety training requirement set by the regulatory authorities before issuing any license to food handlers in Qatar as well as the food inspection conducted by the food health inspectors on a regular basis.Keywords: microbiological quality, street vended food, traditional dishes, Qatar
Procedia PDF Downloads 3131650 Metagenomic Analysis and Pharmacokinetics of Phage Therapy in the Treatment of Bovine Subclinical Mastitis
Authors: Vaibhav D. Bhatt, Anju P. Kunjadia, D. S. Nauriyal, Bhumika J. Joshi, Chaitanya G. Joshi
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Metagenomic analysis of milk samples collected from local cattle breed, kankrej (Bos indicus), Gir (Bos indicus) and Crossbred (Bos indicus X Bos taurus) cattle harbouring subclinical mastitis was carried out by next-generation sequencing (NGS) 454 GS-FLX technology. Around 56 different species including members of Enterobacteriales, Pseudomonadales, Bacillales and Lactobacillales with varying abundance were detected in infected milk. The interesting presence of bacteriophages against Staphylococcus aureus, Escherichia coli, Enterobacter and Yersinia species were observed, especially Enterobacteria and E. coli phages (0∙32%) in Kankrej, Enterobacteria and Staphylococcus phages (1∙05%) in Gir and Staphylococcus phages (2∙32%) in crossbred cattle. NGS findings suggest that phages may be involved in imparting natural resistance of the cattle against pathogens. Further infected milk samples were subjected for bacterial isolation. Fourteen different isolates were identified, and DNA was extracted. Genes (Tet-K, Msr-A, and Mec-A) providing antibiotic resistance to the bacteria were screened by Polymerase Chain Reaction and results were validated with traditional antibiotic assay. Total 3 bacteriophages were isolated from nearby environment of the cattle farm. The efficacy of phages was checked against multi-drug resistant bacteria, identified by PCR. In-vivo study was carried out for phage therapy in mammary glands of female rats “Wister albino”. Mammary glands were infused with MDR isolates for 3 consecutive days. Recovery was observed in infected rats after intramammary infusion of sterile phage suspension. From day 4th onwards, level of C-reactive protein was significant increases up to day 12th . However, significant reduction was observed between days 12th to 18th post treatment. Bacteriophages have significant potential as antibacterial agents and their ability to replicate exponentially within their hosts and their specificity, make them ideal candidates for more sustainable mastitis control.Keywords: bacteriophages, c-reactive protein, mastitis, metagenomic analysis
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