Search results for: escherichia%20coli%20MG%201655

Authors: H. Alijani, M. Jabari, S. Matroodi, H. Zolqarnein, A. Sharafi, I. Zamani

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Actinomycetes, Gram-positive bacteria, are interesting as a main producer of secondary metabolites and are important industrially and pharmaceutically. The marine environment is a potential source for new actinomycetes, which can provide novel bioactive compounds and industrially important enzymes. The aims of this study were to isolate and identify novel actinomycetes from Persian Gulf sediments and screen these isolates for the production of secondary metabolites, especially antibiotics, Using phylogenetic (16S rRNA gene sequence), morphological and biochemical analyses. 15 different actinomycete strains from Persian Gulf sediments at a depth of 5-10 m were identified. DNA extraction was done using Cinnapure DNA Kit. PCR amplification of 16S rDNA gene was performed using F27 and R1492 primers. Phylogenetic tree analysis was performed using the MEGA 6 software. Most of the isolated strains belong to the genus namely Streptomyces (14), followed by Nocardiopsis (1). Antibacterial assay of the isolates supernatant was performed using a standard disc diffusion assay with replication (n=3). The results of disk diffusion assay showed that most active strain against Proteus volgaris and Bacillus cereus was AMJ1 (16.46±0.2mm and 13.78±0.2mm, respectively), against Salmonella sp. AMJ7 was the most effective strain (10.13±0.2mm), and AMJ1 and AHA5 showed more inhibitory activity against Escherichia coli (8.04±0.02 mm and 8.2±0.03 ). The AMJ6 strain showed best antibacterial activity against Klebsiella sp. (8.03±0.02mm). Antifungal activity of AMJ2 showed that it was most active strain against complex (16.05±0.02mm) and against Aspergillus flavus strain AMJ1 was most active strain (16.4±0.2mm) and highest antifungal activity against Trichophyton mentagrophytes, Microsporum gyp serum and Candida albicans, were shown by AHA1 (21.03±0.02mm), AHA3 and AHA7 (18±0.03mm), AMJ6 (21.03±0.2mm) respectively. Our results revealed that the marine actinomycetes of Persian Gulf sediments were potent source of novel antibiotics and bioactive compounds and indicated that the antimicrobial metabolites were extracellular. Most of the secondary metabolites and antibiotics are extracellular in nature and extracellular products of actinomycetes show potent antimicrobial activities.

Keywords: antibacterial activity, antifungal activity, marine actinomycetes, Persian Gulf

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Authors: Sneha Singh, Abhimanyu Dev, Vinod Nigam

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The major issue which decides the impending use of gold nanoparticles (AuNPs) in nanobiotechnological applications is their particle size and stability. Often the AuNPs obtained biomimetically are considered useless owing to their instability in the aqueous medium and thereby limiting the widespread acceptance of this facile green synthesis procedure. So, the use of nontoxic surfactants is warranted to stabilize the biogenic nanoparticles (NPs). But does the surfactant only play a role in stabilizing by being adsorbed to the NPs surface or can it have any other significant contribution in synthesis process and controlling their size as well as shape? Keeping this idea in mind, AuNPs were synthesized by using surfactant treated (lechate) and untreated (cell lysate supernatant) Bacillus licheniformis cell extract. The cell extracts mediated reduction of chloroauric acid (HAuCl 4) in the presence of non-ionic surfactant, Tween 20 (TW20), and its effect on the AuNPs stability was studied. Interestingly, the surfactant used in the study served as potential alternative to harvest cellular enzymes involved in bioreduction process in a hassle free condition. The surfactants ability to solubilize/leach membrane proteins and simultaneously stabilizing the AuNPs could have advantage from process point of view as it will reduce the time and economics involve in the nanofabrication of biogenic NPs. The synthesis was substantiated with UV-Vis spectroscopy, Dynamic light scattering study, FTIR spectroscopy, and Transmission electron microscopy. The Zeta potential of AuNPs solutions was measured routinely to corroborate the stability observations recorded visually. Highly stable, ultra-small AuNPs of 2.6 nm size were obtained from the study. Further, the biological efficacy of the obtained AuNPs as potential antibacterial agent was evaluated against Bacilllus subtilis, Pseudomonas aeruginosa, and Escherichia coli by observing the zone of inhibition. This potential of AuNPs of size < 3 nm as antibacterial agent could pave way for development of new antimicrobials and overcoming the problems of antibiotics resistance

Keywords: antibacterial, bioreduction, nanoparticles, surfactant

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Authors: Adepeju Simon-Oke, Olatunji Odeyemi, Mobolanle Oniya

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Urinary tract infection has become the most common bacterial infections in humans, both at the community and hospital settings; it has been reported in all age groups and in both sexes. This study was carried out in order to determine and evaluate the prevalence, current drug susceptibility pattern of the isolated organisms and identify the associated risk factors of UTIs among the pregnant women in Akure, Ondo State, Nigeria. A cross-sectional study was conducted on the urine of pregnant women, and socio-demographic information of the women was collected. A total of 300 clean midstream urine samples were collected, and a general urine microscopic examination and culture were carried out, the Microbact identification system was used to identify gram-negative bacteria. Out of the 300 urine samples cultured, 183(61.0%) yielded significant growth of urinary pathogens while 117(39.0%) yielded either insignificant growth or no growth of any urinary pathogen. Prevalence of UTI was significantly associated with the type of toilet used, symptoms of UTI, and previous history of urinary tract infection (p<0.05). Escherichia coli 58(31.7%) was the dominant pathogen isolated, and the least isolated uropathogens were Citrobacter freudii and Providencia retgerri 2(1.1%) respectively. Gram-negative bacteria showed 77.6%, 67.9%, and 61.2% susceptibility to ciprofloxacin, augmentin, and chloramphenicol, respectively. Resistance against septrin, chloramphenicol, sparfloxacin, amoxicillin, augmentin, gentamycin, pefloxacin, trivid, and streptomycin was observed in the range of 23.1 to 70.1%. Gram-positive uropathogens isolated showed high resistance to amoxicillin (68.4%) and high susceptibility to the remaining nine antibiotics in the range 65.8% to 89.5%. This study justifies that pregnant women are at high risk of UTI. Therefore screening of pregnant women during antenatal clinics should be considered very important to avoid complications. Health education with regular antenatal and personal hygiene is recommended as precautionary measures to UTI.

Keywords: pregnant women, prevalence, risk factor, UTIs

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Authors: Elham A. Kwildi, Yahia S. Abugnah, Nuri S. Madi

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This study was designed to evaluate the microbiological quality and safety of two types of salads prepared at a catering center affiliated with Libyan Airlines in Tripoli, Libya. Two hundred and twenty-one (221) samples (132 economy-class and 89 first- class) were used in this project which lasted for ten months. Biweekly, microbiological tests were performed which included total plate count (TPC) and total coliforms (TCF), in addition to enumeration and/or detection of some pathogenic bacteria mainly Escherichia coli, Staphylococcus aureus, Bacillus cereus, Salmonella sp, Listeria sp and Vibrio parahaemolyticus parahaemolyticus, By using conventional as well as compact dry methods. Results indicated that TPC of type 1 salad ranged between (<10 – 62 x 103 cfu/gm) and (<10 to 36 x103 cfu/g), while TCF were (<10 – 41 x 103 cfu/gm) and (< 10 to 66 x102 cfu/g) using both methods of detection respectively. On the other hand, TPC of type 2 salad were: (1 × 10 – 52 x 103) and (<10 – 55 x 103 cfu/gm) and in the range of (1 x10 to 45x103 cfu/g), and the (TCF) counts were between (< 10 to 55x103 cfu/g) and (< 10 to 34 x103 cfu/g) using the 1st and the 2nd methods of detection respectively. Also, the pathogens mentioned above were detected in both types of salads, but their levels varied according to the type of salad and the method of detection. The level of Staphylococcus aureus, for instance, was 17.4% using conventional method versus 14.4% using the compact dry method. Similarly, E. coli was 7.6% and 9.8%, while Salmonella sp. recorded the least percentage i.e. 3% and 3.8% with the two mentioned methods respectively. First class salads were also found to contain the same pathogens, but the level of E. coli was relatively higher in this case (14.6% and 16.9%) using conventional and compact dry methods respectively. The second rank came Staphylococcus aureus (13.5%) and (11.2%), followed by Salmonella (6.74%) and 6.70%). The least percentage was for Vibrio parahaemolyticus (4.9%) which was detected in the first class salads only. The other two pathogens Bacillus cereus and Listeria sp. were not detected in either one of the salads. Finally, it is worth mentioning that there was a significant decline in TPC and TCF counts in addition to the disappearance of pathogenic bacteria after the 6-7th month of the study which coincided with the first trial of the HACCP system at the center. The ups and downs in the counts along the early stages of the study reveal that there is a need for some important correction measures including more emphasis on training of the personnel in applying the HACCP system effectively.

Keywords: air travel, vegetable salads, foodborne outbreaks, Libya

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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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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

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Authors: David Sarpong, Waleed Khursheed, Ernest Danquah, Erin Murphy

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Shigella is a pathogenic bacterium responsible for shigellosis, a severe diarrheal disease that claims the lives of immunocompromised individuals worldwide. To develop therapeutics against this disease, an understanding of the molecular mechanisms underlying the pathogen’s physiology is crucial. Small non-coding RNAs (sRNAs) have emerged as important regulators of bacterial physiology, including as components of toxin-antitoxin systems. In this study, we investigated the role of RyfA in S. flexneri physiology and virulence. RyfA, originally identified as an sRNA in Escherichia coli, is conserved within the Enterobacteriaceae family, including Shigella. Whereas two copies of ryfA are present in S. dysenteriae, all other Shigella species contain only one copy of the gene. Additionally, we identified a putative open reading frame within the RyfA transcript, suggesting that it may be a dual-functioning gene encoding a small protein in addition to its sRNA function. To study ryfA in vitro, we cloned the gene into an inducible plasmid and observed the effect on bacterial growth. Here, we report that RyfA production inhibits the growth of S. flexneri, and this inhibition is dependent on the contained open reading frame. In-silico analyses have revealed the presence of two divergently transcribed sRNAs, RyfB1 and RyfB2, which share nucleotide complementarity with RyfA and thus are predicted to function as anti-toxins. Our data demonstrate that RyfB2 has a stronger antitoxin effect than RyfB1. This regulatory pattern suggests a novel form of a toxin-antitoxin system in which the activity of a single toxin is inhibited to varying degrees by two sRNA antitoxins. Studies are ongoing to investigate the regulatory mechanism(s) of the antitoxin genes, as well as the downstream targets and mechanism of growth inhibition by the RyfA toxin. This study offers distinct insights into the regulatory mechanisms underlying Shigella physiology and may inform the development of new anti-Shigella therapeutics.

Keywords: sRNA, shigella, toxin-antitoxin, Type 1 toxin antitoxin

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Authors: M. T. Amina, A. A. Alazba, U. Manzoor

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Solar disinfection (SODIS) is a viable method for household water treatment and is recommended by the World Health Organization as cost effective approach that can be used without special skills. The efficiency of both SODIS and solar collector disinfection (SOCODIS) system was evaluated using four different sources of water including stored rainwater, storm water, ground water and treated sewage. Samples with naturally occurring microorganisms were exposed to sunlight for about 8-9 hours in 2-L polyethylene terephthalate bottles under similar experimental conditions. Total coliform (TC), Escherichia coli (E. coli) and heterotrophic plate counts (HPC) were used as microbial water quality indicators for evaluating the disinfection efficiency at different sunlight intensities categorized as weak, mild and strong weathers. Heterotrophic bacteria showed lower inactivation rates compared to E. coli and TC in both SODIS and SOCODIS system. The SOCODIS system at strong weather was the strongest disinfection system in this study and the complete inactivation of HPC was observed after 8-9 hours of exposure with SODIS being ineffective for HPC. At moderate weathers, however, the SOCODIS system did not show complete inactivation of HPC due to very high concentrations (up to 5x10^7 CFU/ml) in both storm water and treated sewage. SODIS even remained ineffective for the complete inactivation of E. coli due to its high concentrations of about 2.5x10^5 in treated sewage compared with other waters even after 8-9 hours of exposure. At weak weather, SODIS was not effective at all while SOCODIS system, though incomplete, showed good disinfection efficiency except for HPC and to some extent for high E. coli concentrations in storm water. Largest reduction of >5 log occurred for TC when used stored rainwater even after 6 hours of exposure in the case of SOCODIS system at strong weather. The lowest E. coli and HPC reduction of ~2 log was observed in SODIS system at weak weather. Further tests with varying pH and turbidity are required to understand the effects of reaction parameters that could be a step forward towards maximizing the disinfection efficiency of such systems for the complete inactivation of naturally occurring E. coli or HPC at moderate or even at weak weathers.

Keywords: efficiency, microbial, SODIS, SOCODIS, weathers

Procedia PDF Downloads 241

Authors: N. B. Ikenweiwe, A. A. Alimi, N. A. Bamidele, A. O. Ewumi, J. Dairo, I. A. Akinnubi, S. O. Otubusin

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A study on the physical, chemical and biological parameters of the lower course of Ogun River, Isheri-Olofin was carried out between January and December 2014 in order to determine the effects of the anthropogenic activities of the Kara abattoir and domestic waste depositions on the quality of the water. Water samples were taken twice each month at three selected stations A, B and C (based on characteristic features or activity levels) along the water course. Samples were analysed using standard methods for chemical and biological parameters the same day in the laboratory while physical parameters were determined in-situ with water parameters kit. Generally, results of Transparency, Dissolved Oxygen, Nitrates, TDS and Alkalinity fall below the permissible limits of WHO and FEPA standards for drinking and fish production. Results of phosphates, lead and cadmium were also low but still within the permissible limit. Only Temperature and pH were within limit. Low plankton community, (phytoplankton, zooplankton), which ranges from 3, 5 to 40, 23 were as a result of low levels of DO, transparency and phosphate. The presence of coliform bacteria of public health importance like Escherichia coli, Proteus vulgaris, Aeromonas sp., Shigella sp, Enterobacter aerogenes as well as gram negative bacteria Proteus morganii are mainly indicators of faecal pollution. Fish and other resources obtained from this water stand the risk of being contaminated with these organisms and man is at the receiving end. The results of the physical, chemical and some biological parameters of Isheri, Ogun River, according to this study showed that the live forms of aquatic and fisheries resources there are dwelling under stress as a result of deposition of bones, horns, faecal components, slurry of suspended solids, fat and blood into the water. Government should therefore establish good monitoring system against illegal waste depositions and create education programmes that will enlighten the community on the social, ecological and economic values of the river.

Keywords: water parameters, Isheri Ogun river, anthropogenic activities, Kara abattoir

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Authors: Aysenur Ozturk, Aysen Yildiz, Hilal Yilmaz, Pinar Ergenekon, Melek Ozkan

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Water scarcity is one of the most important environmental problems of the World today. Desalination process is regarded as a promising solution to solve drinking water problem of the countries facing with water shortages. Reverse osmosis membranes are widely used for desalination processes. Nano structured biomimetic membrane production is one of the most challenging research subject for improving water filtration efficiency of the membranes and for reducing the cost of desalination processes. There are several researches in the literature on the development of novel biomimetic nanofiltration membranes by incorporation of aquaporin Z molecules. Aquaporins are cell membrane proteins that allow the passage of water molecules and reject all other dissolved solutes. They are present in cell membranes of most of the living organisms and provide high water passage capacity. In this study, GST (Glutathione S-transferas) tagged E. coli aquaporinZ and H. elongate aquaporin proteins, which were previously cloned and characterized, were purified from E. coli BL21 cells and used for fabrication of modified Polysulphone Membrane (PS). Aquaporins were incorporated on the surface of the membrane by using 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) phospolipids as carrier liposomes. Aquaporin containing proteoliposomes were immobilized on the surface of the membrane with m-phenylene-diamine (MPD) and trimesoyl chloride (TMC) rejection layer. Water flux, salt rejection and glucose rejection performances of the thin film composite membranes were tested by using Dead-End Reactor Cell. In this study, effect of proteoliposome concentration, and filtration pressure on water flux and salt rejection rate of membranes were investigated. Type of aquaporin used for membrane fabrication, flux and pressure applied for filtration were found to be important parameters affecting rejection rates. Results suggested that optimization of concentration of aquaporin carriers (proteoliposomes) on the membrane surface is necessary for fabrication of effective composite membranes used for different purposes.

Keywords: aquaporins, biomimmetic membranes, desalination, water treatment

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Authors: Wan-Ling Jiang, Hsin Chi, Jia-Lu Cheng, Ming-Fang Cheng

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Background: Investigating the risk factors for the fecal carriage of third-generation cephalosporin-resistant E.coli could contribute to further disease prevention. Previous research on third-generation cephalosporin-resistant prevalence in children in different regions of Taiwan is limited. This project aims to explore the risk factors and regional differences in the prevalence of third-generation cephalosporin-resistant and other antibiotic-resistant E. coli in the northern, southern, and eastern regions of Taiwan. Methods: We collected data from children aged 0 to 18 from community or outpatient clinics from July 2022 to May 2023 in southern, northern, and eastern Taiwan. The questionnaire was designed to survey the characteristics of participants and possible risk factors, such as clinical information, household environment, drinking water, and food habits. After collecting fecal samples and isolating stool culture with E.coli, antibiotic sensitivity tests and MLST typing were performed. Questionnaires were used to analyze the risk factors of third-generation cephalosporin-resistant E. coli in the three different regions of Taiwan. Results: In the total 246 stool samples, third-generation cephalosporin-resistant E.coli accounted for 37.4% (97/246) of all isolates. Among the three different regions of Taiwan, the highest prevalence of fecal carriage with third-generation cephalosporin-resistant E.coli was observed in southern Taiwan (42.7%), followed by northern Taiwan (35.5%) and eastern Taiwan (28.4%). Multi-drug resistant E. coli had prevalence rates of 51.9%, 66.3%, and 37.1% in the northern, southern, and eastern regions, respectively. MLST typing revealed that ST131 was the most prevalent type (11.8%). The prevalence of ST131 in northern, southern, and eastern Taiwan was 10.1%, 12.3%, and 13.2%, respectively. Risk factors analysis identified lower paternal education, overweight status, and non-vegetarian diet as statistical significance risk factors for third-generation cephalosporin-resistant E.coli. Conclusion: The fecal carriage rates of antibiotic-resistant E. coli among Taiwanese children were on the rise. This study found regional disparities in the prevalence of third-generation cephalosporin-resistant and multi-drug-resistant E. coli, with southern Taiwan having the highest prevalence. Lower paternal education, overweight, and non-vegetarian diet were the potential risk factors of third-generation cephalosporin-resistant E. coli in this study.

Keywords: Escherichia coli, fecal carriage, antimicrobial resistance, risk factors, prevalence

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

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

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

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Authors: Alissa Jane S. Mondejar, Florifern C. Paglinawan, Nanette Hope N. Sumaya, Joey Genevieve T. Martinez, Mylah Villacorte-Tabelin

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Mining is associated with increased heavy metals in the environment, and heavy metal contamination disrupts the activities of soil fauna, such as nematodes, causing changes in the function of the soil ecosystem. Previous studies found that nematode community composition and diversity indices were strongly affected by heavy metals (e.g., Pb, Cu, and Zn). In this study, the influence of heavy metals on nematode survivability and reproduction were investigated. Life history analysis of the free-living nematodes, Heterocephalobellus sp. and Cephalobus sp. (Rhabditida: Cephalobidae) were assessed using the hanging drop technique, a technique often used in life history trait experiments. The nematodes were exposed to different temperatures, i.e.,20°C, 25°C, and 30°C, in different groups (control and heavy metal exposed) and fed with the same bacterial density of 1×109 Escherichia coli cells ml-1 for 30 days. Results showed that increasing temperature and exposure to heavy metals had a significant influence on the survivability and egg production of both species. Heterocephalobellus sp. and Cephalobus sp., when exposed to 20°C survived longer and produced few numbers of eggs but without subsequent hatching. Life history parameters of Heterocephalobellus sp. showed that the value of parameters was higher in the control group under net production rate (R0), fecundity (mx) which is also the same value for the total fertility rate (TFR), generation times (G0, G₁, and Gh) and Population doubling time (PDT). However, a lower rate of natural increase (rm) was observed since generation times were higher. Meanwhile, the life history parameters of Cephalobus sp. showed that the value of net production rate (R0) was higher in the exposed group. Fecundity (mx) which is also the same value for the TFR, G0, G1, Gh, and PDT, were higher in the control group. However, a lower rate of natural increase (rm) was observed since generation times were higher. In conclusion, temperature and exposure to heavy metals had a negative influence on the life history of the nematodes, however, further experiments should be considered.

Keywords: artisanal and small-scale gold mining (ASGM), hanging drop method, heavy metals, life history trait.

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Authors: L. L. Tundisi, A. Pessoa Jr., E. B. Tambourgi, E. Silveira, P. G. Mazzola

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L-asparaginase (L-ASNase) is the gold standard treatment for acute lymphoblastic leukemia that mainly affects pediatric patients; treatment increases survival from 20% to 90%. The characterization of other L-Asparaginases, apart from the most used from Escherichia coli and Erwinia chrysanthemi, has been reported, but the choice of the most appropriate is still under debate. This choice should be based on its pharmacokinetics, immune hypersensitivity, doses, prices, pharmacodynamics. The main factors influencing the antileukemic activity of ASNase are enzymatic activity, Km, glutaminase activity, clearance of the enzyme and development of resistance. However, most of the commercialized enzyme present an intrinsic glutaminase activity, which is responsible for some side effects. In this study, glutaminase free asparaginase produced from Aspergillus oryzae was precipitated in different percentages of ethanol (0–80%), until optimum ethanol concentration of 60% (w/w) was found. Following, precipitation of crude L-ASNase was performed in a single step, using 60% (w/w) ethanol, under constant agitation and temperature. It presented activity of 135.45 U/mg and after gel filtration chromatography with Sephadex G-the enzymatic activity was 322.02 U/mg. The apparent molecular mass of the purified L-ASNase fraction was estimated by 10% SDS-PAGE. Proteins were stained with Coomassie Brilliant Blue R-250. The molar mass range was from 10 kDa to 250 kDa. L-ASNase from Aspergillus oryzae was characterized aiming possible therapeutic use. Four different buffers (phosphate-citrate buffer pH 2.6 to 5.8; phosphate buffer pH 5.8 to 7.4; Tris - HCl pH 7.4 to 9.0; and carbonate buffer pH 9.8 to 10.6) were used to measure the optimum pH for L-ASNase activity. The optimum temperature for enzyme activity was measured at optimal pH conditions (Tris-HCl and phosphate buffer, pH 7.4) at different temperatures ranging from 5 to 55°C. All activities were calculated by quantifying the free ammonia, using the Nessler reagent. The kinetic parameters calculation, e.g. Michaelis-Menten constant (Km), maximum velocity (Vmax) and Hills coefficient (n), were performed by incubating the enzyme in different concentrations of the substrate at optimum conditions of pH and fitted on Hill’s equation. This glutaminase free asparaginase showed a low Km (3.39 mM and 3.81 mM) and enzymatic activity of 135.45 U/mg after precipitation with ethanol. After gel filtration chromatography it rose to 322.02 U/mg. Optimum activity was found between pH 5.8 - 9.0, best activity results with phosphate buffer pH 7.4 and Tris-HCl pH 7.4 and showed activity from 5°C to 55°C. These results indicate that L-ASNase from A. oryzae has the potential for human use.

Keywords: biopharmaceuticals, bioprocessing, bioproducts, biotechnology, enzyme activity, ethanol precipitation

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Authors: Ofonime Ogba, Augustine Akpan

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Objectives and Goals: The study was aimed at determining pathogenic microbial carriage on the external body parts of Camponotus consobrinus which is also known as the banded sugar ant because of its liking for sugar and sweet food. The level of pathogenic microbial carriage of Camponotus consobrinus in association to the environment in which they have been collected is not known. Methods: The ants were purposively collected from four locations including the kitchens, bedroom of various homes, food shops, and bakeries. The sample collection took place within the hours of 6:30 pm to 11:00 pm. The ants were trapped in transparent plastic containers of which sugar, pineapple peels, sugar cane and soft drinks were used as bait. The ants were removed with a sterile spatula and put in 10mls of peptone water in sterile universal bottles. The containers were vigorously shaken to wash the external surface of the ant. It was left overnight and transported to the Microbiology Laboratory, University of Calabar Teaching Hospital for analysis. The overnight peptone broths were inoculated on Chocolate agar, Blood agar, Cystine Lactose Electrolyte-Deficient agar (CLED) and Sabouraud dextrose agar. Incubation was done aerobically and in a carbon dioxide jar for 24 to 48 hours at 37°C. Isolates were identified based on colonial characteristics, Gram staining, and biochemical tests. Results: Out of the 250 Camponotus consobrinus caught for the study, 90(36.0%) were caught in the kitchen, 75(30.0%) in the bedrooms 40(16.0%) in the bakery while 45(18.0%) were caught in the shops. A total of 82.0% prevalence of different microbial isolates was associated with the ants. The kitchen had the highest number of isolates 75(36.6%) followed by the bedroom 55(26.8%) while the bakery recorded the lowest number of isolates 35(17.1%). The profile of micro-organisms associated with Camponotus consobrinus was Escherichia coli 73(30.0%), Morganella morganii 45(18.0%), Candida species 25(10.0%), Serratia marcescens 10(4.0%) and Citrobacter freundii 10(4.0%). Conclusion: Most of the Camponotus consobrinus examined in the four locations harboured potential pathogens. The presence of ants in homes and shops can facilitate the propagation and spread of pathogenic microorganisms. Therefore, the development of basic preventive measures and the control of ants must be taken seriously.

Keywords: Camponotus consobrinus, potential pathogens, microbial isolates, spread

Procedia PDF Downloads 141

Authors: Pampita Chakraborty, Sukumar Mukherjee

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People with diabetes mellitus are more susceptible to developing infections, as high blood sugar levels can weaken the patient's immune system defences. People with diabetes are more adversely affected when they get an infection than someone without the disease, because you have weakened immune defences in diabetes. People who have minimally elevated blood sugar levels experience worse outcomes with infections. Diabetic patients in hospitals do not necessarily have a higher mortality rate due to infections, but they do face longer hospitalisation and recovery times. A study was done in a tertiary care unit in eastern India. Patients with type 2 diabetes mellitus infection were recruited in the study. A total of 520 cases of Type 2 Diabetes Mellitus were recorded out of which 200 infectious cases was included in the study. All subjects underwent detailed history & clinical examination. Microbiological samples were collected from respective site of the infection for microbial culture and antibiotic sensitivity test. Out of the 200 infectious cases urinary tract infection(UTI) was found in majority of the cases followed by diabetic foot ulcer (DFU), respiratory tract infection(RTI) and sepsis. It was observed that Escherichia coli was the most commonest pathogen isolated from UTI cases and Staphylococcus aureus was predominant in foot ulcers followed by other organisms. Klebsiella pneumonia was the major organism isolated from RTI and Enterobacter aerogenes was commonly observed in patients with sepsis. Isolated bacteria showed differential sensitivity pattern against commonly used antibiotics. The majority of the isolates were resistant to several antibiotics that are usually prescribed on an empirical basis. These observations are important, especially for patient management and the development of antibiotic treatment guidelines. It is recommended that diabetic patients receive pneumococcal and influenza vaccine annually to reduce morbidity and mortality. Appropriate usage of antibiotics based on local antibiogram pattern can certainly help the clinician in reducing the burden of infections.

Keywords: antimicrobial resistance, diabetic foot ulcer, respiratory tract infection, urinary tract infection

Procedia PDF Downloads 323

Authors: Holm Zerbe, Laura Macias, Hans-Joachim Schuberth, Wolfram Petzl

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Mastitis is among the most important production diseases in cows. It accounts for large parts of antimicrobial drug use in the dairy industry worldwide. Due to the imminent normative to reduce the use of antimicrobial drugs in livestock, new ways for therapy and prophylaxis of mastitis are needed. Recently epigenetic regulation of inflammation by chromatin modifications has increasingly drawn attention. Currently, some epigenetic modifiers have already been approved for the use in humans, however little is known about their actions in the bovine system. The aim of our study was to investigate whether three selected epigenetic modifiers (Vitamin D3, SAHA and S2101) influence the initial immune response towards mastitis pathogens in bovine udder tissue in vitro. Tissue explants of the teat cistern and udder parenchyma were collected from 21 cows and were incubated for 36 hours in the absence and presence of epigenetic modifiers. Additionally, the tissue was stimulated with heat-inactivated particles of Escherichia coli and Staphylococcus aureus, which are regarded as two of the most important mastitis pathogens. After incubation, the explants were tested by RT-qPCR for transcript abundances of immune-related candidate genes. Gene expression was validated in culture supernatants by an AlphaLISA assay. Furthermore, the culture supernatants were analyzed for their chemotactic capacity through a chemotaxis assay. Statistical analysis of data was performed with the program ‘R’ version 3.2.3. Vitamin D3 had no effect on the immune response of udder tissue in vitro after stimulation with mastitis pathogens. The epigenetic modifiers SAHA and S2101 however significantly blocked the pathogen-induced upregulation of CXCL8, TNFα, S100A9 and LAP (P < 0.05). The regulation of IL10 was not affected by treatment with SAHA and S2101. Transcript abundances for CXCL8 were reflected by IL8 contents and chemotactic activity in culture supernatants. In conclusion, these data show the potential of epigenetic modifiers (SAHA and S2101) to block overshooting inflammation in the udder. Thus epigenetic modifiers may serve in future as immune modulators for the treatment and/or prophylaxis of clinical mastitis. (Funded by Deutsche Forschungsgemeinschaft PE 1495/2-1).

Keywords: mastitis, cattle, epigenetics, immunomodulation

Procedia PDF Downloads 209

Authors: Nimet Yildirim Tirgil, Ahmed Busnaina, April Z. Gu

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Environmental waters are monitored worldwide to protect people from infectious diseases primarily caused by enteric pathogens. All long, Escherichia coli (E. coli) is a good indicator for potential enteric pathogens in waters. Thus, a rapid and simple detection method for E. coli is very important to predict the pathogen contamination. In this study, to the best of our knowledge, as the first time we developed a rapid, direct and reusable SWCNTs (single walled carbon nanotubes) based biosensor system for sensitive and selective E. coli detection in water samples. We use a novel and newly developed flexible biosensor device which was fabricated by high-rate nanoscale offset printing process using directed assembly and transfer of SWCNTs. By simple directed assembly and non-covalent functionalization, aptamer (biorecognition element that specifically distinguish the E. coli O157:H7 strain from other pathogens) based SWCNTs biosensor system was designed and was further evaluated for environmental applications with simple and cost-effective steps. The two gold electrode terminals and SWCNTs-bridge between them allow continuous resistance response monitoring for the E. coli detection. The detection procedure is based on competitive mode detection. A known concentration of aptamer and E. coli cells were mixed and after a certain time filtered. The rest of free aptamers injected to the system. With hybridization of the free aptamers and their SWCNTs surface immobilized probe DNA (complementary-DNA for E. coli aptamer), we can monitor the resistance difference which is proportional to the amount of the E. coli. Thus, we can detect the E. coli without injecting it directly onto the sensing surface, and we could protect the electrode surface from the aggregation of target bacteria or other pollutants that may come from real wastewater samples. After optimization experiments, the linear detection range was determined from 2 cfu/ml to 10⁵ cfu/ml with higher than 0.98 R² value. The system was regenerated successfully with 5 % SDS solution over 100 times without any significant deterioration of the sensor performance. The developed system had high specificity towards E. coli (less than 20 % signal with other pathogens), and it could be applied to real water samples with 86 to 101 % recovery and 3 to 18 % cv values (n=3).

Keywords: aptamer, E. coli, environmental detection, nanobiosensor, SWCTs

Procedia PDF Downloads 170

Authors: Carles Ferré, Ferran Llopis, Javier Jacob, Jordi Giol, Xavier Palom, Ignasi Bardés

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Objective: To analyze the characteristics of acute bacterial prostatitis (ABP) in elderly patients attended in the emergency department (ED). Methods: Observational and cohort study with prospective follow-up including patients with ABP presenting to the ED from January-December 2012. Data were collected for demographic variables, comorbidities, clinical and microbiological findings, treatment, outcome, and reconsultation at 30 days follow up. Findings were compared between patients ≥ 75 years (study group) and < 75 years (control group). Results: During the study period 241 episodes of ABP were included for analysis. Mean age was 62,9 ± 16 years, and 64 (26.5%) were ≥ 75 years old. A history of prostate adenoma was reported in 54 cases (22,4%), diabetes mellitus in 47 patients (19,5%) and prior manipulation of the lower urinary tract in 40 (17%). Mean symptoms duration was 3.38 ± 4.04 days, voiding symptoms were present in 176 cases (73%) and fever in 154 (64%). From 216 urine cultures, 128 were positive (59%) and 24 (17,6%) out of 136 blood cultures. Escherichia coli was the main pathogen in 58.6% of urine cultures and 64% of blood cultures (with resistant strains to fluoroquinolones in 27,7%, cotrimoxazole in 22,9% and amoxicillin/clavulanic in 27.7% of cases). Seventy patients (29%) were admitted to the hospital, and 3 died. At 30-day follow-up, 29 patients (12%) returned to the ED. In the bivariate analysis previous manipulation of the urinary tract, history of cancer, previous antibiotic treatment, resistant E. coli strains to amoxicillin-clavulanate and ciprofloxacin and extended spectrum beta-lactamase (ESBL) producers, renal impairment, and admission to the hospital were significantly more frequent (p < 0.05) among patients ≥ 75 years compared to those younger than 75 years. Conclusions: Ciprofloxacin and amoxicillin-clavulanate appear not to be good options for the empiric treatment of ABP for patients ≥ 75 years given the drug-resistance pattern in our series, and the proportion of ESBL-producing strains of E. coli should be taken into account. Awaiting bacteria identification and antibiogram from urine and/or blood cultures, treatment on an inpatient basis should be considered in older patients with ABP.

Keywords: acute bacterial prostatitits, antibiotic resistance, elderly patients, emergency

Procedia PDF Downloads 351

Authors: Monika Ksiezopolska-Gocalska, Pawel Albrycht, Robert Holyst

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Surface Enhanced Raman Spectroscopy (SERS) is a technique used to analyze very low concentrations of substances in solutions, even in aqueous solutions - which is its advantage over IR. This technique can be used in the pharmacy (to check the purity of products); forensics (whether at a crime scene there were any illegal substances); or medicine (serving as a medical test) and lots more. Due to the high potential of this technique, its increasing popularity in analytical laboratories, and simultaneously - the absence of appropriate platforms enhancing the SERS signal (crucial to observe the Raman effect at low analyte concentration in solutions (1 ppm)), we decided to invent our own SERS platforms. As an enhancing layer, we have chosen gold and silver nanoparticles, because these two have the best SERS properties, and each has an affinity for the other kind of particles, which increases the range of research capabilities. The next step was to commercialize them, which resulted in the creation of the company ‘SERSitive.eu’ focusing on production of highly sensitive (Ef = 10⁵ – 10⁶), homogeneous and reproducible (70 - 80%) substrates. SERStive SERS substrates are made using the electrodeposition of silver or silver-gold nanoparticles technique. Thanks to a very detailed analysis of data based on studies optimizing such parameters as deposition time, temperature of the reaction solution, applied potential, used reducer, or reagent concentrations using a standardized compound - p-mercaptobenzoic acid (PMBA) at a concentration of 10⁻⁶ M, we have developed a high-performance process for depositing precious metal nanoparticles on the surface of ITO glass. In order to check a quality of the SERSitive platforms, we examined the wide range of the chemical compounds and the biological substances. Apart from analytes that have great affinity to the metal surfaces (e.g. PMBA) we obtained very good results for those fitting less the SERS measurements. Successfully we received intensive, and what’s more important - very repetitive spectra for; amino acids (phenyloalanine, 10⁻³ M), drugs (amphetamine, 10⁻⁴ M), designer drugs (cathinone derivatives, 10⁻³ M), medicines and ending with bacteria (Listeria, Salmonella, Escherichia coli) and fungi.

Keywords: nanoparticles, Raman spectroscopy, SERS, SERS applications, SERS substrates, SERSitive

Procedia PDF Downloads 132

Authors: Ridwaan N. Milase, Leonardo J. Van Zyl, Marla Trindade

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American foulbrood (AFB) is the world’s most devastating honeybee disease that has drastically reduced the population of Apis mellifera capensis since 2009. The outbreak has jeopardized the South African bee keeping industry as well as the agricultural sector dependent on honeybees for honey production and pollination, leading to significant economic losses. AFB is caused by Paenibacillus larvae, a spore-forming, Gram positive facultative anaerobic and flagellated bacterium. The use of antibiotics within beehives has selected for resistant strains of P. larvae, while the current practice of burning spore contaminated beehives and equipment contributes to the economic losses in the honeybee-keeping industry. Therefore, phage therapy is proposed as a promising alternative to combat P. larvae strains affecting A. mellifera capensis. The genomes of two P. larvae strains isolated from infected combs in the Western Cape have been sequenced and annotated using bioinformatics tools. Genome analyses has revealed that these P. larvae strains are lysogens to more than 6 different prophages and possess different type of clustered regularly interspaced short palindromic repeat (CRISPRs) regions per strain. Active prophages from one of the two P. larvae strains were detected and identified using PCR. Electron microscopy was used to determine the family of the identified active prophages. Lytic bacteriophages that specifically target the two P. larvae strains were purified from sewage wastewater, beehive materials, and soil samples to investigate their potential development as anti-P. larvae agents. Another alternative treatment being investigated is the development of a prophage endolysin cocktail. Endolysin genes of the prophages have been targeted, cloned and expressed in Escherichia coli. The heterologously expressed endolysins have been purified and are currently being assessed for their lytic activity against P. larvae strains and other commensal microorganisms that compose the honeybee larvae microbiota. The study has shown that phage therapy and endolysins have a great potential as alternative control methods for AFB disease affecting A. mellifera capensis.

Keywords: American foulbrood, bacteriophage, honeybee, Paenibacillus larvae

Procedia PDF Downloads 157

Authors: Mohammed Sabbah, Asmaa Al-Asmar, Doaa Abu-Hani, Fuad Al-Rimawi

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Plant extracts are an increasingly popular natural product with a wide range of potential applications in food, industrial, and health care industries. They are rich in polyphenolic compounds and flavonoids, which have been demonstrated to possess a variety of beneficial properties, including antimicrobial and antioxidant activity. Plant extracts have been found to possess antimicrobial activity against a variety of foodborne pathogens and can be used as a natural preservative to extend the shelf life of food products. They have also strong antioxidant activity, which can reduce the formation of free radicals and oxidation of food components. Recently there is an increase interest in bio-based polymers to be used as innovative “bioplastics” for industrial exploitation e.g. packaging materials for food products. Additionally, incorporation of active compounds (e.g. antioxidants and antimicrobials) in bio-polymer materials is of particular interest since such active polymers can be used as active packaging materials (with antimicrobial and antioxidant activity). In this work, different plant extracts have been characterized for their phenolic compounds, flavonoids content, antioxidant activity (both as free radical scavenging ability and reducing ability), and antimicrobial activity against gram positive and negative bacteria (Escherichia coli; Staphylococcus aureus, and Pseudomonas aeruginosa) as well as antifungal activities (against yeast, mold and Botrytis cinera/a plant pathogen). Results showed that many extracts are rich with polyphenolic compounds and flavonoids and have strong antioxidant activities, and rich with phytochemicals (e.g. rutin, quercetin, oleuropein, tyrosol and hydroxytyrosol). Some extracts showed antibacterial activity against both gram positive and negative bacteria as well as antifungal activities and can work, therefore, as preservatives for food or pharmaceutical industries. As an application, two extracts were used as additive to pectin-based packaging film, and results showed that the addition of these extracts significantly improve their functionality as antimicrobial and antioxidant activity. These biomaterials, therefore can be used in food packaging materials to extend the shelf life of food products.

Keywords: plant extracts, antioxidants, flavonoids, bioplastic, edible biofilm, packaging materials

Procedia PDF Downloads 53

Authors: J. Nyenhuis, J. Drelich

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This research study was designed to determine if food crops could be bio-fortified with micro-nutrients by growing sprouts on mineral fortified fiber mats. Diets high in processed foods have been found to lack essential micro-nutrients for optimum human development and overall health. Some micro-nutrients such as copper (Cu) have been found to enhance the inflammatory response through its oxidative functions, thereby having a role in cardiovascular disease (CVD), metabolic syndrome (MetS), diabetes and related complications. Recycled cellulose fibers and clay saturated with micro-nutrient ions can be converted to a novel mineral-metal hybrid material in which the fiber mat becomes a carrier of essential micro-nutrients. The reduction of ionic to metallic copper was accomplished using hydrogen at temperatures ranging from 400o to 600oC. Copper particles with diameters ranging from ~1 to 400-500 nm reside on the recycled fibers that make up the mats. Seeds purchased from a commercial, organic supplier were germinated on the specially engineered cellulose fiber mats that incorporated w10 wt% clay fillers saturated with either copper particles or ionic copper. After the appearance of the first leaves, the sprouts were dehydrated and analyzed for Cu content. Nutrient analysis showed 1.5 to 1.6 increase in Cu of the sprouts grown on the fiber mats with copper particles, and 2.3 to 2.5 increase on mats with ionic copper as compared to the control samples. The antibacterial properties of materials saturated with copper ions at room temperature and at temperatures up to 400°C have been verified with halo method tests against Escherichia Coli in previous studies. E. coli is a known pathogenic risk in sprout production. Copper exhibits excellent antibacterial properties when tested on S. aureus, a pathogenic gram-positive bacterium. This has also been confirmed for the fiber-copper hybrid material in this study. This study illustrates the potential for the use of engineered mats as a viable way to increase the micro-nutrient composition of locally-grown food crops and the need for additional research to determine the uptake, nutritional implications and risks of micro-nutrient bio-fortification.

Keywords: bio-fortification, copper nutrient analysis, micro-nutrient uptake, sprouts and mineral-fortified mats

Procedia PDF Downloads 330

Authors: Michail Milas, Aikaterini Dafni Tegiou, Nickolas Rigopoulos, Eustathios Giaouris, Zaharias Loannou

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Nanotechnology, a relatively new scientific field, addresses the manipulation of nanoscale materials and devices, which are governed by unique properties, and is applied in a wide range of industries, including food packaging. The incorporation of nanoparticles into polymer matrices used for food packaging is a field that is highly researched today. One such combination is silver nanoparticles with polypropylene. In the present study, the synthesis of the silver nanoparticles was carried out by a natural method. In particular, a ripe banana peel extract was used. This method is superior to others as it stands out for its environmental friendliness, high efficiency and low-cost requirement. In particular, a 1.75 mM AgNO₃ silver nitrate solution was used, as well as a BPE concentration of 1.7% v/v, an incubation period of 48 hours at 70°C and a pH of 4.3 and after its preparation, the polypropylene films were soaked in it. For the PP films, random PP spheres were melted at 170-190°C into molds with 0.8cm diameter. This polymer was chosen as it is suitable for plastic parts and reusable plastic containers of various types that are intended to come into contact with food without compromising its quality and safety. The antimicrobial test against Escherichia coli DFSNB1 and Staphylococcus epidermidis DFSNB4 was performed on the films. It appeared that the films with silver nanoparticles had a reduction, at least 100 times, compared to those without silver nanoparticles, in both strains. The limit of detection is the lower limit of the vertical error lines in the presence of nanoparticles, which is 3.11. The main reasons that led to the adsorption of nanoparticles are the porous nature of polypropylene and the adsorption capacity of nanoparticles on the surface of the films due to hydrophobic-hydrophilic forces. The most significant parameters that contributed to the results of the experiment include the following: the stage of ripening of the banana during the preparation of the plant extract, the temperature and residence time of the nanoparticle solution in the oven, the residence time of the polypropylene films in the nanoparticle solution, the number of nanoparticles inoculated on the films and, finally, the time these stayed in the refrigerator so that they could dry and be ready for antimicrobial treatment.

Keywords: antimicrobial control, banana peel extract, E. coli, natural synthesis, microbe, plant extract, polypropylene films, S.epidermidis, silver nano, random pp

Procedia PDF Downloads 149

Authors: Sze Wing Ho, Nagendra P. Shah

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Lactic acid bacteria (LAB) have shown the beneficial effects on human gastrointestinal tract, such as protects diarrhea induced by lactose intolerance or enteric pathogens. Citrulline is a non-protein amino acid and also the precursors of arginine and nitric oxide, it has shown to enhance intestinal barrier function. Citrulline has shown to improve the growth of some strains of LAB, it is important for LAB to have a sufficient cell concentration to contribute the effects. Therefore, the aims of this study were to investigate the effect of combining citrulline with LAB on the anti-adhesion effect against pathogens and the effect on the cell integrity. The effect of citrulline on selected LAB was determined by incubating in 0%, 0.1% or 0.2% citrulline enriched MRS broth for 18 h. The adhesion ability of LAB and the anti-adhesion effect of LAB and citrulline against pathogens were performed on IPEC-J2 cell line. Transepithelial electrical resistance (TEER) assay was used to measure the tight junction (TJ) integrity. TJ proteins (claudin-1, occludin and zonula occluden-1 (ZO-1)) were determined by western blot analysis. It found that the growth of Lactobacillus helveticus ASCC 511 was significantly stimulated by 0.2% citrulline compared with control during 18 h fermentation. The adhesion of L. helveticus ASCC 511 and Lactobacillus delbrueckii ssp. bulgaricus (L. bulgaricus) ASCC 756 was increased when supplemented with citrulline. Citrulline has shown significant inhibitory effect on the adhesion of Escherichia coli PELI0480 (O157:H7), Shigella sonnei ATCC 25931, Staphyloccocus aureus CMCC26003 and Cronobacter sakazakii ATCC 29544. The anti-adhesion effect of L. helveticus ASCC 511, L. bulgaricus ASCC 756 and Lactobacillus paracasei ASCC 276 against Cronobacter sakazakii ATCC 29544 was significantly enhanced with citrulline supplementation. Treatments with citrulline and LAB were able to maintain the TEER of IPEC-J2 cell and shown the positive effect on the TJ proteins. In conclusion, citrulline had stimulating effect on some strains of LAB and determined to improve the adhesion of LAB on intestinal epithelial cell, to enhance the inhibitory effect on enteric pathogens adhesion as well as had beneficial effects on maintaining cell integrity. It implied LAB supplemented with citrulline might have advantageous effects on gastrointestinal tracts.

Keywords: citrulline, lactic acid bacteria, amino acid, anti-adhesion effect, cell integrity

Procedia PDF Downloads 221

Authors: Ade O. Oyewole, Sunday O. Okoh, Ruth O. Ishola, Adenike D. Odusote, Chima C. Igwe, Gloria N. Elemo, Anthony I. Okoh

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Azadirachta indica (Neem tree) also referred to as an all-purpose tree is used in a wide range of medical preparations in tropical and subtropical countries for prevention and management of various livestock, crops products and human diseases. In Nigeria however, the potentials of this plant have not been fully exploited thus it causes an environmental nuisance during the fruiting season. With a rise in the demand for herbal personal care products globally extracts from different parts of the neem plant were used as the bio-active ingredients in the formulation of personal care products. In this study, formulated neem soap, body cream, lotion, toothpaste and shampoo are analyzed to determine their antibacterial, antifungal, and toxicity properties. The efficacies of these products for management of infectious diseases, both oral and dermal, were also investigated in vitro. Oil from the neem seeds obtained using a mechanical press and acetone extracts of both the neem bark and leaves obtained by the maceration method were used in the formulation and production of the neem personal care products. The antimicrobial and toxicity properties of these products were investigated by agar diffusion, and haemolytic methods respectively. The five neem products (NPs) exhibited strong antibacterial activities against four multi–drug resistant pathogenic and three none pathogenic bacterial strains (Escherichia coli (180), Listeria ivanovii, Staphylococcus aureus, Enterobacter cloacae, Vibro spp., Streptococcus uberis, Mycobacterium smegmatis), except the neem lotion with insignificant activity against E. coli and S. aureus. The minimum inhibitory concentration (MIC) range was between 0.20-0.40 mg/ mL. The 5 NPs demonstrated moderate activity against three clinical dermatophytes isolates (Tinea corporis, Tinea capitis, and Tinea cruiz) as well as one fungal strain (Candida albican) with the MIC ranging between 0.30 - 0.50 mg/ mL and 0.550 mg/mL respectively. The soap and shampoo were the most active against test bacteria and fungi. The haemolytic analysis results on the 5 NPs indicated none toxicity at 0.50 mg/ mL in sheep red blood cells (SRBC).

Keywords: antimicrobial, Azadirachta indica, multi–drug resistant pathogenic bacteria, personal care products

Procedia PDF Downloads 243

Authors: Naira Sahakyan, Margarit Petrosyan, Armen Trchounian

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One of the tasks in contemporary biotechnology, pharmacology and other fields of human activities is to obtain biologically active substances from plants. They are very essential in the treatment of many diseases due to their actually high therapeutic value without visible side effects. However, sometimes the possibility of obtaining the metabolites is limited due to the reduction of wild-growing plants. That is why the plant cell cultures are of great interest as alternative sources of biologically active substances. Besides, during the monitored cultivation, it is possible to obtain substances that are not synthesized by plants in nature. Isolated culture of Ajuga genevensis with high growth activity and ability of regeneration was obtained using MS nutrient medium. The agar-diffusion method showed that aqueous extracts of callus culture revealed high antimicrobial activity towards various gram-positive (Bacillus subtilis A1WT; B. mesentericus WDCM 1873; Staphylococcus aureus WDCM 5233; Staph. citreus WT) and gram-negative (Escherichia coli WKPM M-17; Salmonella typhimurium TA 100) microorganisms. The broth dilution method revealed that the minimal and half maximal inhibitory concentration values against E. coli corresponded to the 70 μg/mL and 140 μg/mL concentration of the extract respectively. According to the photochemiluminescent analysis, callus tissue extracts of leaf and root origin showed higher antioxidant activity than the same quantity of A. genevensis intact plant extract. A. genevensis intact plant and callus culture extracts showed no cytotoxic effect on K-562 suspension cell line of human chronic myeloid leukemia. The GC-MS analysis showed deep differences between the qualitative and quantitative composition of callus culture and intact plant extracts. Hexacosane (11.17%); n-hexadecanoic acid (9.33%); and 2-methoxy-4-vinylphenol (4.28%) were the main components of intact plant extracts. 10-Methylnonadecane (57.0%); methoxyacetic acid, 2-tetradecyl ester (17.75%) and 1-Bromopentadecane (14.55%) were the main components of A. genevensis callus culture extracts. Obtained data indicate that callus culture of A. genevensis can be used as an alternative source of biologically active substances.

Keywords: Ajuga genevensis, antibacterial activity, antioxidant activity, callus cultures

Procedia PDF Downloads 276

Authors: Igbakura I. Luga, Alex A. Adikwu

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A comparison of resistance to quinolones was carried out on isolates of Shiga toxin-producing Escherichia coliO157:H7 from cattle and mecA and nuc genes harbouring Staphylococcus aureus from pigs. The isolates were separately tested in the first and current decades of the 21^st century. The objective was to demonstrate the dissemination of resistance to this frontline class of antibiotic by bacteria from food animals and bring to the limelight the spread of antibiotic resistance in Nigeria. A total of 10 isolates of the E. coli O157:H7 and 9 of mecA and nuc genes harbouring S. aureus were obtained following isolation, biochemical testing, and serological identification using the Remel Wellcolex E. coli O157:H7 test. Shiga toxin-production screening in the E. coli O157:H7 using the verotoxin E. coli reverse passive latex agglutination (VTEC-RPLA) test; and molecular identification of the mecA and nuc genes in S. aureus. Detection of the mecA and nuc genes were carried out using the protocol by the Danish Technical University (DTU) using the following primers mecA-1:5'-GGGATCATAGCGTCATTATTC-3', mecA-2: 5'-AACGATTGTGACACGATAGCC-3', nuc-1: 5'-TCAGCAAATGCATCACAAACAG-3', nuc-2: 5'-CGTAAATGCACTTGCTTCAGG-3' for the mecA and nuc genes, respectively. The nuc genes confirm the S. aureus isolates and the mecA genes as being methicillin-resistant and so pathogenic to man. The fluoroquinolones used in the antibiotic resistance testing were norfloxacin (10 µg) and ciprofloxacin (5 µg) in the E. coli O157:H7 isolates and ciprofloxacin (5 µg) in the S. aureus isolates. Susceptibility was tested using the disk diffusion method on Muller-Hinton agar. Fluoroquinolone resistance was not detected from isolates of E. coli O157:H7 from cattle. However, 44% (4/9) of the S. aureus were resistant to ciprofloxacin. Resistance of up to 44% in isolates of mecA and nuc genes harbouring S. aureus is a compelling evidence for the rapid spread of antibiotic resistance from bacteria in food animals from Nigeria. Ciprofloxacin is the drug of choice for the treatment of Typhoid fever, therefore widespread resistance to it in pathogenic bacteria is of great public health significance. The study concludes that antibiotic resistance in bacteria from food animals is on the increase in Nigeria. The National Food and Drug Administration and Control (NAFDAC) agency in Nigeria should implement the World Health Organization (WHO) global action plan on antimicrobial resistance. A good starting point can be coordinating the WHO, Office of International Epizootics (OIE), Food and Agricultural Organization (FAO) tripartite draft antimicrobial resistance monitoring and evaluation (M&E) framework in Nigeria.

Keywords: Fluoroquinolone, Nigeria, resistance, Staphylococcus aureus

Procedia PDF Downloads 432

Authors: Kichan Lee, Kwang-Ho Choi, Mi-Hye Hwang, Young Min Son, Bang-Hun Hyun, Byeong Yeal Jung

Abstract:

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 323

Authors: Aasaithambi Kalaiselvi, Michael Gabriel Paulraj, Ekambaram Nakkeeran

Abstract:

Emergences of mosquito vector-borne diseases are considered as a perpetual problem globally in tropical countries. The outbreak of several diseases such as chikungunya, zika virus infection and dengue fever has created a massive threat towards the living population. Frequent usage of synthetic insecticides like Dichloro Diphenyl Trichloroethane (DDT) eventually had its adverse harmful effects on humans as well as the environment. Since there are no perennial vaccines, prevention, treatment or drugs available for these pathogenic vectors, WHO is more concerned in eradicating their breeding sites effectively without any side effects on humans and environment by approaching plant-derived natural eco-friendly bio-insecticides. The aim of this study is to investigate the larvicidal potency of Cinnamomum verum essential oil (CEO) loaded niosomes. Cholesterol and surfactant variants of Span 20, 60 and 80 were used in synthesizing CEO loaded niosomes using Transmembrane pH gradient method. The synthesized CEO loaded niosomes were characterized by Zeta potential, particle size, Fourier Transform Infrared Spectroscopy (FT-IR), GC-MS and SEM analysis to evaluate charge, size, functional properties, the composition of secondary metabolites and morphology. The Z-average size of the formed niosomes was 1870.84 nm and had good stability with zeta potential -85.3 meV. The entrapment efficiency of the CEO loaded niosomes was determined by UV-Visible Spectrophotometry. The bio-potency of CEO loaded niosomes was treated and assessed against gram-positive (Bacillus subtilis) and gram-negative (Escherichia coli) bacteria and fungi (Aspergillus fumigatus and Candida albicans) at various concentrations. The larvicidal activity was evaluated against II to IV instar larvae of Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus at various concentrations for 24 h. The mortality rate of LC₅₀ and LC₉₀ values were calculated. The results exhibited that CEO loaded niosomes have greater efficiency against mosquito larvicidal activity. The results suggest that niosomes could be used in various applications of biotechnology and drug delivery systems with greater stability by altering the drug of interest.

Keywords: Cinnamomum verum, niosomes, entrapment efficiency, bactericidal and fungicidal, mosquito larvicidal activity

Procedia PDF Downloads 144