Search results for: antimicrobial preservative
Commenced in January 2007
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Edition: International
Paper Count: 864

Search results for: antimicrobial preservative

84 An Efficient and Low Cost Protocol for Rapid and Mass in vitro Propagation of Hyssopus officinalis L.

Authors: Ira V. Stancheva, Ely G. Zayova, Maria P. Geneva, Marieta G. Hristozkova, Lyudmila I. Dimitrova, Maria I. Petrova

Abstract:

The study describes a highly efficient and low-cost protocol for rapid and mass in vitro propagation of medicinal and aromatic plant species (Hyssopus officinalis L., Lamiaceae). Hyssop is an important aromatic herb used for its medicinal values because of its antioxidant, anti-inflammatory and antimicrobial properties. The protocol for large-scale multiplication of this aromatic plant was developed using young stem tips explants. The explants were sterilized with 0.04% mercuric chloride (HgCl₂) solution for 20 minutes and washing three times with sterile distilled water in 15 minutes. The cultural media was full and half strength Murashige and Skoog medium containing indole-3-butyric acid. Full and ½ Murashige and Skoog media without auxin were used as controls. For each variant 20 glass tubes with two plants were used. In each tube two tip and nodal explants were inoculated. Maximum shoot and root number were obtained on ½ Murashige and Skoog medium supplemented with 0.1 mg L-1 indole-3-butyric acid at the same time after four weeks of culture. The number of shoots per explant and shoot height were considered. The data on rooting percentage, the number of roots per plant and root length were collected after the same cultural period. The highest percentage of survival 85% for this medicinal plant was recorded in mixture of soil, sand and perlite (2:1:1 v/v/v). This mixture was most suitable for acclimatization of all propagated plants. Ex vitro acclimatization was carried out at 24±1 °C and 70% relative humidity under 16 h illuminations (50 μmol m⁻²s⁻¹). After adaptation period, the all plants were transferred to the field. The plants flowered within three months after transplantation. Phenotypic variations in the acclimatized plants were not observed. An average of 90% of the acclimatized plants survived after transferring into the field. All the in vitro propagated plants displayed normal development under the field conditions. Developed in vitro techniques could provide a promising alternative tool for large-scale propagation that increases the number of homologous plants for field cultivation. Acknowledgments: This study was conducted with financial support from National Science Fund at the Bulgarian Ministry of Education and Science, Project DN06/7 17.12.16.

Keywords: Hyssopus officinalis L., in vitro culture, micro propagation, acclimatization

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83 Green Synthesis of Nanosilver-Loaded Hydrogel Nanocomposites for Antibacterial Application

Authors: D. Berdous, H. Ferfera-Harrar

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Superabsorbent polymers (SAPs) or hydrogels with three-dimensional hydrophilic network structure are high-performance water absorbent and retention materials. The in situ synthesis of metal nanoparticles within polymeric network as antibacterial agents for bio-applications is an approach that takes advantage of the existing free-space into networks, which not only acts as a template for nucleation of nanoparticles, but also provides long term stability and reduces their toxicity by delaying their oxidation and release. In this work, SAP/nanosilver nanocomposites were successfully developed by a unique green process at room temperature, which involves in situ formation of silver nanoparticles (AgNPs) within hydrogels as a template. The aim of this study is to investigate whether these AgNPs-loaded hydrogels are potential candidates for antimicrobial applications. Firstly, the superabsorbents were prepared through radical copolymerization via grafting and crosslinking of acrylamide (AAm) onto chitosan backbone (Cs) using potassium persulfate as initiator and N,N’-methylenebisacrylamide as the crosslinker. Then, they were hydrolyzed to achieve superabsorbents with ampholytic properties and uppermost swelling capacity. Lastly, the AgNPs were biosynthesized and entrapped into hydrogels through a simple, eco-friendly and cost-effective method using aqueous silver nitrate as a silver precursor and curcuma longa tuber-powder extracts as both reducing and stabilizing agent. The formed superabsorbents nanocomposites (Cs-g-PAAm)/AgNPs were characterized by X-ray Diffraction (XRD), UV-visible Spectroscopy, Attenuated Total reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR), Inductively Coupled Plasma (ICP), and Thermogravimetric Analysis (TGA). Microscopic surface structure analyzed by Transmission Electron Microscopy (TEM) has showed spherical shapes of AgNPs with size in the range of 3-15 nm. The extent of nanosilver loading was decreased by increasing Cs content into network. The silver-loaded hydrogel was thermally more stable than the unloaded dry hydrogel counterpart. The swelling equilibrium degree (Q) and centrifuge retention capacity (CRC) in deionized water were affected by both contents of Cs and the entrapped AgNPs. The nanosilver-embedded hydrogels exhibited antibacterial activity against Escherichia coli and Staphylococcus aureus bacteria. These comprehensive results suggest that the elaborated AgNPs-loaded nanomaterials could be used to produce valuable wound dressing.

Keywords: antibacterial activity, nanocomposites, silver nanoparticles, superabsorbent Hydrogel

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82 Application of Metarhizium anisopliae against Meloidogyne javanica in Soil Amended with Oak Debris

Authors: Mohammad Abdollahi

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Tomato (Lycopersicon esculentum Mill.) is one of the most popular, widely grown and the second most important vegetable crop, after potatoes. Nematodes have been identified as one of the major pests affecting tomato production throughout the world. The most destructive nematodes are the genus Meloidogyne. Most widespread and devastating species of this genus are M. incognita, M. javanica, and M. arenaria. These species can cause complete crop loss under adverse growing conditions. There are several potential methods for management of the root knot nematodes. Although the chemicals are widely used against the phytonematodes, because of hazardous effects of these compounds on non-target organisms and on the environment, there is a need to develop other control strategies. Nowadays, non-chemical measures are widely used to control the plant parasitic nematodes. Biocontrol of phytonematodes is an important method among environment-friendly measures of nematode management. There are some soil-inhabiting fungi that have biocontrol potential on phytonematodes, which can be used in nematode management program. The fungus Metarhizium anisopliae, originally is an entomopathogenic bioagent. Biocontrol potential of this fungus on some phytonematodes has been reported earlier. Recently, use of organic soil amendments as well as the use of bioagents is under special attention in sustainable agriculture. This research aimed to reduce the pesticide use in control of root-knot nematode, Meloidogyne javanica in tomato. The effects of M. anisopliae IMI 330189 and different levels of oak tree debris on M. javanica were determined. The combination effect of the fungus as well as the different rates of soil amendments was determined. Pots were filled with steam pasteurized soil mixture and the six leaf tomato seedlings were inoculated with 3000 second stage larvae of M. javanica/kg of soil. After eight weeks, plant growth parameters and nematode reproduction factors were compared. Based on the results of our experiment, combination of M. anisopliae IMI 330189 and oak debris caused more than 90% reduction in reproduction factor of nematode, at the rates of 100 and 150 g/kg soil (P ≤ 0.05). As compared to control, the reduction in number of galls was 76%. It was 86% for nematode reproduction factor, showing the significance of combined effect of both tested agents. Our results showed that plant debris can increase the biological activity of the tested bioagent. It was also proved that there was no adverse effect of oak debris, which potentially has antimicrobial activity, on antagonistic power of applied bioagent.

Keywords: biological control, nematode management, organic soil, Quercus branti, root knot nematode, soil amendment

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81 Post-Harvest Biopreservation of Fruit and Vegetables with Application of Lactobacillus Strains

Authors: Judit Perjessy, Zsolt Zalan, Ferenc Hegyi, Eniko Horvath-Szanics, Krisztina Takacs, Andras Nagy, Adel Klupacs, Erika Koppany-Szabo, Zhirong Wang, Kaituo Wang, Muying Du, Jianquan Kan

Abstract:

The post-harvest diseases cause great economic losses in the fruit and vegetables; the prevention of these deterioration has great importance. Against the fungi, which cause most of the diseases, are extensively used the fungicides. However, there are increasing consumer concerns over the presence of pesticide residues in food. An alternative and in recent years, increasingly studied method for the prevention of the diseases is biocontrol, where antagonistic microorganisms are used for the control of fungi. The genera of Lactobacillus is well known and extensively studied, but its applicability as biocontrol agents in post-harvest preservation of fruit and vegetables is poorly investigated. However these bacteria can be found on the surface of the plants and have great antimicrobial activity. In our study we have investigated the chitinase activity, the antifungal effect and the applicability of several Lactobacillus strains to select potential biocontrol agents. We investigated the determination of the environmental parameters of a gene (encoding chitinase) expression and we also investigated the relationship between actual antifungal activity and potential chitinase activity. Mixed cultures were also developed to enhance the antifungal activity and determined the optimal mold spore and bacteria concentration ratio for the appropriate efficacy. Five Lactobacillus strains (L. acidophilus N2, L. delbrueckii subsp. bulgaricus B397, L. sp. 2231, L. sake subsp. sake 2471, L. buchneri 1145) possess chitinase-coding gene from the 43 investigated Lactobacillus strains. Proteins with similar molecular weight and separation properties like bacterial chitinases were detected from these strains, which also possess chitin-binding property. Nevertheless, they were inactive, lacks the chitinolytic activity. In point of the cumulative activity of inhibition, our results showed that certain strains were statistically significant in a positive direction compared to other strains, e.g., L. rhamnosus VT1 and L. Casey 154 have shown great general antifungal effect against 11 molds from the genera Penicillium and Botrytis and isolated from spoiled fruit and vegetables. Also, some mixed cultures (L. rhamnosus VT1 - L. Plantarum 299v) showed significant antifungal effects against the indigenous molds on the surface of apple fruit during the industrial storage experiment. Thus, they could be promising for post-harvest biopreservation.

Keywords: biocontrol, chitinase, Lactobacillus, post-harvest

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80 Antimicrobial Resistance: Knowledge towards Antibiotics in a Mexican Population

Authors: L. D. Upegui, Isabel Alvarez-Solorza, Karina Garduno-Ulloa, Maren Boecker

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Introduction: The increasing prevalence rate of resistant and multiresistant bacterial strains to antibiotics is a threat to public health and requires a rapid multifunctional answer. Individuals that are affected by resistant strains present a higher morbidity and mortality than individuals that are infected with the same species of bacteria but with sensitive strains. There have been identified risk factors that are related to the misuse and overuse of antibiotics, like socio-demographic characteristics and psychological aspects of the individuals that have not been explored objectively due to a lack of valid and reliable instruments for their measurement. Objective: To validate a questionnaire for the evaluation of the levels of knowledge related to the use of antibiotics in a Mexican population. Materials and Methods: Analytical cross-sectional observational study. The questionnaire consists of 12 items to evaluated knowledge (1=no, 2=not sure, 3=yes) regarding the use of antibiotics, with higher scores corresponding to a higher level of knowledge. Data are collected in a sample of students. Data collection is still ongoing. In this abstract preliminary results of 30 respondents are reported which were collected during pilot-testing. The validation of the instrument was done using the Rasch model. Fit to the Rasch model was tested checking overall fit to the model, unidimensionality, local independence and evaluating the presence of Differential Item Functioning (DIF) by age and gender. The software Rumm2030 and the SPSS were used for the analyses. Results: The participants of the pilot-testing presented an average age of 32 years ± 12.6 and 53% were women. The preliminary results indicated that the items showed good fit to the Rasch model (chi-squared=12.8 p=0.3795). Unidimensionality (number of significant t-tests of 3%) could be proven, the items were locally independent, and no DIF was observed. Knowledge was the smallest regarding statements on the role of antibiotics in treating infections, e.g., most of the respondents did not know that antibiotics would not work against viral infections (70%) and that they could also cause side effects (87%). The knowledge score ranged from 0 to 100 points with a transformed measurement (mean of knowledge 27.1 ± 4.8). Conclusions: The instrument showed good psychometric proprieties. The low scores of knowledge about antibiotics suggest that misinterpretations on the use of these medicaments were prevalent, which could influence the production of antibiotic resistance. The application of this questionnaire will allow the objective identification of 'Hight risk groups', which will be the target population for future educational campaigns, to reduce the knowledge gaps on the general population as an effort against antibiotic resistance.

Keywords: antibiotics, knowledge, misuse, overuse, questionnaire, Rasch model, validation

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79 Antimicrobial Activity, Phytochemistry and Toxicity Of Extracts Of Naturally Growing and Cultivated Aloe Turkanensis

Authors: Zachary Muthii Rukenya, James Mbaria, Peter Mbaabu, Kiama Stephen Gitahi, Ronald Onzago

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Aloe turkanensis is one of the widely used medicinal shrub and in Kenya the plant is mainly found in Baringo, Isiolo, Laikipia, Turkana and West Pokot Counties where it is used in ethno-medicine and ethno-veterinary medicine. The Turkana community uses the plant products to manage malaria, wounds, stomach ache, constipation, pain, skin infection, poultry diseases and retained afterbirth in cows. This evaluated the efficacy and safety of the plant obtained from Turkana County, Kenya. Preliminary data on the use of the plant in the county was collected through observation, photographing and interviews. A sample of the whole plant was harvested in Natira sublocation, in ex-Turkana west district in February 2012 after identification by Aloe-working group herbalists who voluntarily provided information on its medicinal uses. Botanical identification was done at Kenya Forest Research Centre in Karura where voucher specimen was deposited. Cold maceration using 70% methanol and distilled water was used for extraction. Bioassays were to determine the effects of the plant extracts on brine shrimp and selected bacterial and fungal cultures. The extracts were tested in-vitro activity against standard cultures of B. cereus (ATCC 11778), S. aureus (ATCC25923), P. aeroginosa (ATCC 27853), E. coli (ATCC 25922) and a human infections clinical isolate of C. albicans. The extracts of Aloe turkanensis inhibited the growth B. cereus (100-200 mg/ml), S. aureus (50-100 mg/ml), P. aeroginosa (200mg/ml), E. coli (400mg/ml) while C. albicans was not affected. The extracts also inhibited the growth of S. aureus and B. cereus with mean diameters of inhibition zones being 19.75±1 mm and 18.5±05 mm reapectively. Phytochemical screening showed the presence of alkaloids, tarpenoids, steroids, quinones, saponins and tannins in the plant extracts. The extract was found to be non-toxic at a concentration of 1000µg/ml with a 100% survival of Brine Shrimp larva. It was concluded that Aloe turkanensis growing the study area has metabolites that inhibit the growth of microorganisms and is however, there is need for further studies to validate the in-vivo bioactivity of the plant and more generate adequate toxicological data.to support conservation, value chain addition of its products and widespread use as a herbal remedy.

Keywords: Aloe turkanensis, bioactivity, cultivated, human infections

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78 The Role of Microbes in Organic Sustainable Agriculture and Plant Protection

Authors: Koppula Prawan, Kehinde D. Oyeyemi, Kushal P. Singh

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As people become more conscious of the detrimental consequences of conventional agricultural practices on the environment and human health, organic, sustainable agriculture and plant protection employing microorganisms have grown in importance. Although the use of microorganisms in agriculture is a centuries-old tradition, it has recently attracted renewed interest as a sustainable alternative to chemical-based plant protection and fertilization. Healthy soil is the cornerstone of sustainable agriculture, and microbes are essential to this process. Synthetic fertilizers and pesticides can destroy the beneficial microorganisms in the soil, upsetting the ecosystem's equilibrium. By utilizing organic farming's natural practices, such as the usage of microbes, it aims to maintain and improve the health of the soil. Microbes have several functions in agriculture, including nitrogen fixation, phosphorus solubilization, and disease suppression. Nitrogen fixation is the process by which certain microbes, such as rhizobia and Azotobacter, convert atmospheric nitrogen into a form that plants can use. Phosphorus solubilization involves the conversion of insoluble phosphorus into a soluble form that plants can absorb. Disease suppression involves the use of microbes to control plant diseases by competing with pathogenic organisms for resources or by producing antimicrobial compounds. Microbes can be applied to plants through seed coatings, foliar sprays, or soil inoculants. Seed coatings involve applying a mixture of microbes and nutrients to the surface of seeds before planting. Foliar sprays involve applying microbes and nutrients to the leaves of plants during the growing season. Soil inoculants involve adding microbes to the soil before planting. The use of microbes in plant protection and fertilization has several advantages over conventional methods. Firstly, microbes are natural and non-toxic, making them safe for human health and the environment. Secondly, microbes have the ability to adapt to changing environmental conditions, making them more resilient to drought and other stressors. Finally, the use of microbes can reduce the need for synthetic fertilizers and pesticides, reducing costs and minimizing environmental impact. In conclusion, organic, sustainable agriculture and plant protection using microbes are an effective and sustainable alternatives to conventional farming practices. The use of microbes can help to preserve and enhance soil health, increase plant productivity, and reduce the need for synthetic fertilizers and pesticides. As the demand for organic and sustainable agriculture continues to grow, the use of microbes is likely to become more widespread, providing a more environmentally friendly and sustainable future for agriculture.

Keywords: microbes, inoculants, fertilization, soil health, conventional.

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77 Green Synthesis of Silver and Silver-Gold Alloy Nanoparticle Using Cyanobacteria as Bioreagent

Authors: Piya Roychoudhury, Ruma Pal

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Cyanobacteria, commonly known as blue green algae were found to be an effective bioreagent for nanoparticle synthesis. Nowadays silver nanoparticles (AgNPs) are very popular due to their antimicrobial and anti-proliferative activity. To exploit these characters in different biotechnological fields, it is very essential to synthesize more stable, non-toxic nano-silver. For this reason silver-gold alloy (Ag-AuNPs) nanoparticles are of great interest as they are more stable, harder and more effective than single metal nanoparticles. In the present communication we described a simple technique for rapid synthesis of biocompatible AgNP and Ag-AuNP employing cyanobacteria, Leptolyngbya and Lyngbya respectively. For synthesis of AgNP the biomass of Leptolyngbya valderiana (200 mg Fresh weight) was exposed to 9 mM AgNO3 solution (pH 4). For synthesis of Ag-AuNP Lyngbya majuscula (200 mg Fresh weight) was exposed to equimolar solution of hydrogen tetra-auro chlorate and silver nitrate (1mM, pH 4). After 72 hrs of exposure thallus of Leptolyngyba turned brown in color and filaments of Lyngbya turned pink in color that indicated synthesis of nanoparticles. The produced particles were extracted from the cyanobacterial biomass using nano-capping agent, sodium citrate. Firstly, extracted brown and pink suspensions were taken for Energy Dispersive X-ray (EDAX) analysis to confirm the presence of silver in brown suspension and presence of both gold and silver in pink suspension. Extracted nanoparticles showed a distinct single plasmon band (AgNP at 411 nm; Ag-Au NP at 481 nm) in Uv-vis spectroscopy. It was revealed from Transmission electron microscopy (TEM) that all the synthesized particles were spherical in nature with a size range of ~2-25 nm. In X-ray powder diffraction (XRD) analysis four intense peaks appeared at 38.2°, 44.5°, 64.8°and 77.8° which confirmed the crystallographic nature of synthesized particles. Presence of different functional groups viz. N-H, C=C, C–O, C=O on the surface of nanoparticles were recorded by Fourier transform infrared spectroscopy (FTIR). Scanning Electron microscopy (SEM) images showed the surface topography of metal treated filaments of cyanobacteria. The stability of the particles was observed by Zeta potential study. Antibiotic property of synthesized particles was tested by Agar well diffusion method against gram negative bacteria Pseudomonas aeruginosa. Overall, this green-technique requires low energy, less manufacturing cost and produces rapidly eco-friendly metal nanoparticles.

Keywords: cyanobacteria, silver nanoparticles, silver-gold alloy nanoparticles, spectroscopy

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76 Preparation of Allyl BODIPY for the Click Reaction with Thioglycolic Acid

Authors: Chrislaura Carmo, Luca Deiana, Mafalda Laranjo, Abilio Sobral, Armando Cordova

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Photodynamic therapy (PDT) is currently used for the treatment of malignancies and premalignant tumors. It is based on the capture of a photosensitizing molecule (PS) which, when excited by light at a certain wavelength, reacts with oxygen and generates oxidizing species (radicals, singlet oxygen, triplet species) in target tissues, leading to cell death. BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s-indaceno) derivatives are emerging as important candidates for photosensitizer in photodynamic therapy of cancer cells due to their high triplet quantum yield. Today these dyes are relevant molecules in photovoltaic materials and fluorescent sensors. In this study, it will be demonstrated the possibility that BODIPY can be covalently linked to thioglycolic acid through the click reaction. Thiol−ene click chemistry has become a powerful synthesis method in materials science and surface modification. The design of biobased allyl-terminated precursors with high renewable carbon content for the construction of the thiol-ene polymer networks is essential for sustainable development and green chemistry. The work aims to synthesize the BODIPY (10-(4-(allyloxy) phenyl)-2,8-diethyl-5,5-difluoro-1,3,7,9-tetramethyl-5H-dipyrrolo[1,2-c:2',1'-f] [1,3,2] diazaborinin-4-ium-5-uide) and to click reaction with Thioglycolic acid. BODIPY was synthesized by the condensation reaction between aldehyde and pyrrole in dichloromethane, followed by in situ complexation with BF3·OEt2 in the presence of the base. Then it was functionalized with allyl bromide to achieve the double bond and thus be able to carry out the click reaction. The thiol−ene click was performed using DMPA (2,2-Dimethoxy-2-phenylacetophenone) as a photo-initiator in the presence of UV light (320–500 nm) in DMF at room temperature for 24 hours. Compounds were characterized by standard analytical techniques, including UV-Vis Spectroscopy, 1H, 13C, 19F NMR and mass spectroscopy. The results of this study will be important to link BODIPY to polymers through the thiol group offering a diversity of applications and functionalization. This new molecule can be tested as third-generation photosensitizers, in which the dye is targeted by antibodies or nanocarriers by cells, mainly in cancer cells, PDT and Photodynamic Antimicrobial Chemotherapy (PACT). According to our studies, it was possible to visualize a click reaction between allyl BODIPY and thioglycolic acid. Our team will also test the reaction with other thiol groups for comparison. Further, we will do the click reaction of BODIPY with a natural polymer linked with a thiol group. The results of the above compounds will be tested in PDT assays on various lung cancer cell lines.

Keywords: bodipy, click reaction, thioglycolic acid, allyl, thiol-ene click

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75 Development and Validation of a Green Analytical Method for the Analysis of Daptomycin Injectable by Fourier-Transform Infrared Spectroscopy (FTIR)

Authors: Eliane G. Tótoli, Hérida Regina N. Salgado

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Daptomycin is an important antimicrobial agent used in clinical practice nowadays, since it is very active against some Gram-positive bacteria that are particularly challenges for the medicine, such as methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE). The importance of environmental preservation has receiving special attention since last years. Considering the evident need to protect the natural environment and the introduction of strict quality requirements regarding analytical procedures used in pharmaceutical analysis, the industries must seek environmentally friendly alternatives in relation to the analytical methods and other processes that they follow in their routine. In view of these factors, green analytical chemistry is prevalent and encouraged nowadays. In this context, infrared spectroscopy stands out. This is a method that does not use organic solvents and, although it is formally accepted for the identification of individual compounds, also allows the quantitation of substances. Considering that there are few green analytical methods described in literature for the analysis of daptomycin, the aim of this work was the development and validation of a green analytical method for the quantification of this drug in lyophilized powder for injectable solution, by Fourier-transform infrared spectroscopy (FT-IR). Method: Translucent potassium bromide pellets containing predetermined amounts of the drug were prepared and subjected to spectrophotometric analysis in the mid-infrared region. After obtaining the infrared spectrum and with the assistance of the IR Solution software, quantitative analysis was carried out in the spectral region between 1575 and 1700 cm-1, related to a carbonyl band of the daptomycin molecule, and this band had its height analyzed in terms of absorbance. The method was validated according to ICH guidelines regarding linearity, precision (repeatability and intermediate precision), accuracy and robustness. Results and discussion: The method showed to be linear (r = 0.9999), precise (RSD% < 2.0), accurate and robust, over a concentration range from 0.2 to 0.6 mg/pellet. In addition, this technique does not use organic solvents, which is one great advantage over the most common analytical methods. This fact contributes to minimize the generation of organic solvent waste by the industry and thereby reduces the impact of its activities on the environment. Conclusion: The validated method proved to be adequate to quantify daptomycin in lyophilized powder for injectable solution and can be used for its routine analysis in quality control. In addition, the proposed method is environmentally friendly, which is in line with the global trend.

Keywords: daptomycin, Fourier-transform infrared spectroscopy, green analytical chemistry, quality control, spectrometry in IR region

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74 Molecular Detection of Staphylococcus aureus in the Pork Chain Supply and the Potential Anti-Staphylococcal Activity of Natural Compounds

Authors: Valeria Velasco, Ana M. Bonilla, José L. Vergara, Alcides Lofa, Jorge Campos, Pedro Rojas-García

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Staphylococcus aureus is both commensal bacterium and opportunistic pathogen that can cause different diseases in humans and can rapidly develop antimicrobial resistance. Since this bacterium has the ability to colonize the nares and skin of humans and animals, there is a risk of contamination of food in different steps of the food chain supply. Emerging strains have been detected in food-producing animals and meat, such as methicillin-resistant S. aureus (MRSA). The aim of this study was to determine the prevalence and oxacillin susceptibility of S. aureus in the pork chain supply in Chile and to suggest some natural antimicrobials for control. A total of 487 samples were collected from pigs (n=332), carcasses (n=85), and retail pork meat (n=70). Presumptive S. aureus colonies were isolated by selective enrichment and culture media. The confirmation was carried out by biochemical testing (Api® Staph) and molecular technique PCR (detection of nuc and mecA genes, associated with S. aureus and methicillin resistance, respectively). The oxacillin (β-lactam antibiotic that replaced methicillin) susceptibility was assessed by minimum inhibitory concentration (MIC) using the Epsilometer test (Etest). A preliminary assay was carried out to test thymol, carvacrol, oregano essential oil (Origanum vulgare L.), Maqui or Chilean wineberry extract (Aristotelia chilensis (Mol.) Stuntz) as anti-staphylococcal agents using the disc diffusion method at different concentrations. The overall prevalence of S. aureus in the pork chain supply reached 33.9%. A higher prevalence of S. aureus was determined in carcasses (56.5%) than in pigs (28.3%) and pork meat (32.9%) (P ≤ 0.05). The prevalence of S. aureus in pigs sampled at farms (40.6%) was higher than in pigs sampled at slaughterhouses (23.3%) (P ≤ 0.05). The contamination of no packaged meat with S. aureus (43.1%) was higher than in packaged meat (5.3%) (P ≤ 0.05). The mecA gene was not detected in S. aureus strains isolated in this study. Two S. aureus strains exhibited oxacillin resistance (MIC ≥ 4µg/mL). Anti-staphylococcal activity was detected in solutions of thymol, carvacrol, and oregano essential oil at all concentrations tested. No anti-staphylococcal activity was detected in Maqui extract. Finally, S. aureus is present in the pork chain supply in Chile. Although the mecA gene was not detected, oxacillin resistance was found in S. aureus and could be attributed to another resistance mechanism. Thymol, carvacrol, and oregano essential oil could be used as anti-staphylococcal agents at low concentrations. Research project Fondecyt No. 11140379.

Keywords: antimicrobials, mecA gen, nuc gen, oxacillin susceptibility, pork meat

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73 Evaluation of Medicinal Plants, Catunaregam spinosa, Houttuynia cordata, and Rhapis excelsa from Malaysia for Antibacterial, Antifungal and Antiviral Properties

Authors: Yik Sin Chan, Bee Ling Chuah, Wei Quan Chan, Ri Jin Cheng, Yan Hang Oon, Kong Soo Khoo, Nam Weng Sit

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Traditionally, medicinal plants have been used to treat different kinds of ailments including infectious diseases. They serve as a good source of lead compounds for the development of new and safer anti-infective agents. This study aimed to investigate the antimicrobial potential of the leaves of three medicinal plants, namely Catunaregam spinosa (Rubiaceae; Mountain pomegranate), Houttuynia cordata (Saururaceae; "fishy-smell herb") and Rhapis excelsa (Arecaceae; “broadleaf lady palm”). The leaves extracts were obtained by sequential extraction using hexane, chloroform, ethyl acetate, ethanol, methanol and water. The antibacterial and antifungal activities were assessed using a colorimetric broth microdilution method against a panel of human pathogenic bacteria (Gram-positive: Bacillus cereus and Staphylococcus aureus; Gram-negative: Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa) and fungi (yeasts: Candida albicans, Candida parapsilosis and Cryptococcus neoformans; Moulds: Aspergillus fumigatus and Trichophyton mentagrophytes) respectively; while antiviral activity was evaluated against the Chikungunya virus on monkey kidney epithelial (Vero) cells by neutral red uptake assay. All the plant extracts showed bacteriostatic activity, however, only 72% of the extracts (13/18) were found to have bactericidal activity. The lowest minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were given by the hexane extract of C. spinosa against S. aureus with the values of 0.16 and 0.31 mg/mL respectively. All the extracts also possessed fungistatic activity. Only the hexane, chloroform and ethyl acetate extracts of H. cordata exerted inhibitory activity against A. fumigatus, giving the lowest fungal susceptibility index of 16.7%. In contrast, only 61% of the extracts (11/18) showed fungicidal activity. The ethanol extract of R. excelsa exhibited the strongest fungicidal activity against C. albicans, C. parapsilosis and T. mentagrophytes with minimum fungicidal concentration (MFC) values of 0.04–0.08 mg/mL, in addition to its methanol extract against T. mentagrophytes (MFC=0.02 mg/mL). For anti-Chikungunya virus activity, only chloroform and ethyl acetate extracts of R. excelsa showed significant antiviral activity with 50% effective concentrations (EC50) of 29.9 and 78.1 g/mL respectively. Extracts of R. excelsa warrant further investigations into their active principles responsible for antifungal and antiviral properties.

Keywords: bactericidal, Chikungunya virus, extraction, fungicidal

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72 Ecofriendly Synthesis of Au-Ag@AgCl Nanocomposites and Their Catalytic Activity on Multicomponent Domino Annulation-Aromatization for Quinoline Synthesis

Authors: Kanti Sapkota, Do Hyun Lee, Sung Soo Han

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Nanocomposites have been widely used in various fields such as electronics, catalysis, and in chemical, biological, biomedical and optical fields. They display broad biomedical properties like antidiabetic, anticancer, antioxidant, antimicrobial and antibacterial activities. Moreover, nanomaterials have been used for wastewater treatment. Particularly, bimetallic hybrid nanocomposites exhibit unique features as compared to their monometallic components. Hybrid nanomaterials not only afford the multifunctionality endowed by their constituents but can also show synergistic properties. In addition, these hybrid nanomaterials have noteworthy catalytic and optical properties. Notably, Au−Ag based nanoparticles can be employed in sensor and catalysis due to their characteristic composition-tunable plasmonic properties. Due to their importance and usefulness, various efforts were developed for their preparation. Generally, chemical methods have been described to synthesize such bimetallic nanocomposites. In such chemical synthesis, harmful and hazardous chemicals cause environmental contamination and increase toxicity levels. Therefore, ecologically benevolent processes for the synthesis of nanomaterials are highly desirable to diminish such environmental and safety concerns. In this regard, here we disclose a simple, cost-effective, external additive free and eco-friendly method for the synthesis of Au-Ag@AgCl nanocomposites using Nephrolepis cordifolia root extract. Au-Ag@AgCl NCs were obtained by the simultaneous reduction of cationic Ag and Au into AgCl in the presence of plant extract. The particle size of 10 to 50 nm was observed with the average diameter of 30 nm. The synthesized nanocomposite was characterized by various modern characterization techniques. For example, UV−visible spectroscopy was used to determine the optical activity of the synthesized NCs, and Fourier transform infrared (FT-IR) spectroscopy was employed to investigate the functional groups present in the biomolecules that were responsible for both reducing and capping agents during the formation of nanocomposites. Similarly, powder X-ray diffraction (XRD), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), thermogravimetric analysis (TGA) and energy-dispersive X-ray (EDX) spectroscopy were used to determine crystallinity, size, oxidation states, thermal stability and weight loss of the synthesized nanocomposites. As a synthetic application, the synthesized nanocomposite exhibited excellent catalytic activity for the multicomponent synthesis of biologically interesting quinoline molecules via domino annulation-aromatization reaction of aniline, arylaldehyde, and phenyl acetylene derivatives. Interestingly, the nanocatalyst was efficiently recycled for five times without substantial loss of catalytic properties.

Keywords: nanoparticles, catalysis, multicomponent, quinoline

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71 Bacteriophage Is a Novel Solution of Therapy Against S. aureus Having Multiple Drug Resistance

Authors: Sanjay Shukla, A. Nayak, R. K. Sharma, A. P. Singh, S. P. Tiwari

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Excessive use of antibiotics is a major problem in the treatment of wounds and other chronic infections, and antibiotic treatment is frequently non-curative, thus alternative treatment is necessary. Phage therapy is considered one of the most promising approaches to treat multi-drug resistant bacterial pathogens. Infections caused by Staphylococcus aureus are very efficiently controlled with phage cocktails, containing a different individual phages lysate infecting a majority of known pathogenic S. aureus strains. The aim of the present study was to evaluate the efficacy of a purified phage cocktail for prophylactic as well as therapeutic application in mouse model and in large animals with chronic septic infection of wounds. A total of 150 sewage samples were collected from various livestock farms. These samples were subjected for the isolation of bacteriophage by the double agar layer method. A total of 27 sewage samples showed plaque formation by producing lytic activity against S. aureus in the double agar overlay method out of 150 sewage samples. In TEM, recovered isolates of bacteriophages showed hexagonal structure with tail fiber. In the bacteriophage (ØVS) had an icosahedral symmetry with the head size 52.20 nm in diameter and long tail of 109 nm. Head and tail were held together by connector and can be classified as a member of the Myoviridae family under the order of Caudovirale. Recovered bacteriophage had shown the antibacterial activity against the S. aureus in vitro. Cocktail (ØVS1, ØVS5, ØVS9, and ØVS 27) of phage lysate were tested to know in vivo antibacterial activity as well as the safety profile. Result of mice experiment indicated that the bacteriophage lysate were very safe, did not show any appearance of abscess formation, which indicates its safety in living system. The mice were also prophylactically protected against S. aureus when administered with cocktail of bacteriophage lysate just before the administration of S. aureuswhich indicates that they are good prophylactic agent. The S. aureusinoculated mice were completely recovered by bacteriophage administration with 100% recovery, which was very good as compere to conventional therapy. In the present study, ten chronic cases of the wound were treated with phage lysate, and follow up of these cases was done regularly up to ten days (at 0, 5, and 10 d). The result indicated that the six cases out of ten showed complete recovery of wounds within 10 d. The efficacy of bacteriophage therapy was found to be 60% which was very good as compared to the conventional antibiotic therapy in chronic septic wounds infections. Thus, the application of lytic phage in single dose proved to be innovative and effective therapy for the treatment of septic chronic wounds.

Keywords: phage therapy, S aureus, antimicrobial resistance, lytic phage, and bacteriophage

Procedia PDF Downloads 94
70 Endemic Asteraceae from Mauritius Islands as Potential Phytomedicines

Authors: S.Kauroo, J. Govinden Soulange, D. Marie

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Psiadia species from the Asteraceae are traditionally used in the folk medicine of Mauritius to treat cutaneous and bronchial infections. The present study aimed at validating the phytomedicinal properties of the selected species from the Asteraceae family, namely Psiadia arguta, Psiadia viscosa, Psiadia lithospermifolia, and Distephanus populifolius. Dried hexane, ethyl acetate, and methanol leaf extracts were studied for their antioxidant properties using the DPPH (1, 1-diphenyl-2-picryl-hydrazyl), FRAP (Ferric Reducing Ability of Plasma), and Deoxyribose assays. Antibacterial activity against human pathogenic bacteria namely Escherichia coli (ATCC 27853), Staphylococcus aureus (ATCC 29213), Enterococcus faecalis (ATCC 29212), Klebsiella pneumonia (ATCC27853), Pseudomonas aeruginosa (ATCC 27853), and Bacillus cereus (ATCC 11778) was measured using the broth microdilution assay. Qualitative phytochemical screening using standard methods revealed the presence of coumarins, tannins, leucoanthocyanins, and steroids in all the tested extracts. The measured phenolics level of the selected plant extracts varied from 24.0 to 231.6 mg GAE/g with the maximum level in methanol extracts in all four species. The highest flavonoids and proanthocyanidins content was noted in Psiadia arguta methanolic extracts with 65.7±1.8 mg QE/g and 5.1±0.0 mg CAT/g dry weight (DW) extract, respectively. The maximum free radical scavenging activity was measured in Psiadia arguta methanol and ethyl acetate extracts with IC50 11.3±0.2 and 11.6± 0.2 µg/mL, respectively and followed by Distephanus populifolius methanol extracts with an IC50 of 11.3± 0.8 µg/mL. The maximum ferric reducing antioxidant potential was noted in Psiadia lithospermifolia methanol extracts with a FRAP value of 18.8 ± 0.4 µmol Fe2+/L/g DW. The antioxidant capacity based on DPPH and Deoxyribose values were negatively related to total phenolics, flavonoid and proanthocyanidins content while the ferric reducing antioxidant potential were strongly correlated to total phenolics, flavonoid and proanthocyanidins content. All four species exhibited antimicrobial activity against the tested bacteria (both Gram-negative and Gram-positive). Interestingly, the hexane and ethyl acetate extracts of Psiadia viscosa and Psiadia lithospermifolia were more active than the control antibiotic Chloramphenicol. The Minimum inhibitory concentration (MIC) values for hexane and ethyl acetate extracts of Psiadia viscosa and Psiadia lithospermifolia against the tested bacteria ranged from (62.5 to 500 µg/ml). These findings validate the use of these tested Asteraceae in the traditional medicine of Mauritius and also highlight their pharmaceutical potential as prospective phytomedicines.

Keywords: antibacterial, antioxidant, DPPH, flavonoids, FRAP, Psiadia spp

Procedia PDF Downloads 499
69 Cytotoxic Effect of Biologically Transformed Propolis on HCT-116 Human Colon Cancer Cells

Authors: N. Selvi Gunel, L. M. Oktay, H. Memmedov, B. Durmaz, H. Kalkan Yildirim, E. Yildirim Sozmen

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Object: Propolis which consists of compounds that are accepted as antioxidant, antimicrobial, antiseptic, antibacterial, anti-inflammatory, anti-mutagenic, immune-modulator and cytotoxic, is frequently used in current therapeutic applications. However, some of them result in allergic side effects, causing consumption to be restricted. Previously our group has succeeded in producing a new biotechnological product which was less allergenic. In this study, we purpose to optimize production conditions of this biologically-transformed propolis and determine the cytotoxic effects of obtained new products on colon cancer cell line (HCT-116). Method: Firstly, solid propolis samples were dissolved in water after weighing, grinding and sizing (sieve-35mesh) and applied 40 kHz/10 min ultrasonication. Samples were prepared according to inoculation with Lactobacillus plantarum in two different proportions (2.5% and 3.5%). Chromatographic analyzes of propolis were performed by UPLC-MS/MS (Waters, Milford, MA) system. Results were analysed by UPLC-MS/MS system MassLynx™ 4.1 software. HCT-116 cells were treated with propolis examples at 25-1000 µg/ml concentrations and cytotoxicity were measured by using WST-8 assay at 24, 48, and 72 hours. Samples with biological transformation were compared with the non-transformed control group samples. Our experiment groups were formed as follows: untreated (group 1), propolis dissolved in water ultrasonicated at 40 kHz/10 min (group 2), propolis dissolved in water ultrasonicated at 40 kHz/10 min and inoculated 2.5% L. plantarum L1 strain (group 3), propolis dissolved in water ultrasonicated at 40 kHz/10 min and inoculated 3.5% L. plantarum L3 strain (group 4). Obtained data were calculated with Graphpad Software V5 and analyzed by two-way ANOVA test followed by Bonferroni test. Result: As a result of our study, the cytotoxic effect of propolis samples on HCT-116 cells was evaluated. There was a 7.21 fold increase in group 3 compared to group 2 in the concentration of 1000 µg/ml, and it was a 6.66 fold increase in group 3 compared to group 1 at the end of 24 hours. At the end of 48 hours, in the concentration of 500 µg/ml, it was determined 4.7 fold increase in group 4 compared to group 3. At the same time, in the concentration of 750 µg/ml it was determined 2.01 fold increase in group 4 compared to group 3 and in the same concentration, it was determined 3.1 fold increase in group 4 compared to group 2. Also, at the 72 hours, in the concentration of 750 µg/ml, it was determined 2.42 fold increase in group 3 according to group 2 and in the same time, in the concentration of 1000 µg/ml, it was determined 2.13 fold increase in group 4 according to group 2. According to cytotoxicity results, the group which were ultrasonicated at 40 kHz/10min and inoculated 3.5% L. plantarum L3-strain had a higher cytotoxic effect. Conclusion: It is known that bioavailability of propolis is halved in six months. The data obtained from our results indicated that biologically-transformed propolis had more cytotoxic effect than non-transformed group on colon cancer cells. Consequently, we suggested that L. plantarum-transformation provides both reduction of allergenicity and extension of bioavailability period by enhancing healthful polyphenols.

Keywords: bio-transformation, propolis, colon cancer, cytotoxicity

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68 Bacteriological Spectrum and Resistance Patterns of Common Clinical Isolates from Infections in Cancer Patients

Authors: Vivek Bhat, Rohini Kelkar, Sanjay Biswas

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Introduction: Cancer patients are at increased risk of bacterial infections. This may due to the disease process itself, the effect of chemotherapeutic drugs or invasive procedures such as catheterization. A wide variety of bacteria including some emerging pathogens are increasingly being reported from these patients. The incidence of multidrug-resistant organisms particularly in the Gram negative group is also increasing, with higher resistance rates seen to cephalosporins, β-lactam/β-lactam inhibitor combinations, and the carbapenems. This study documents the bacteriological spectrum of infections and their resistance patterns in cancer patients. Methods: This study includes all bacterial isolates recovered from infections cancer patients over a period of 18 months. Samples included Blood cultures, Pus/wound swabs, urine, tissue biopsies, body fluids, catheter tips and respiratory specimens such as sputum and bronchoalveolar lavage (BAL). All samples were processed in the microbiology laboratory as per standard laboratory protocols. Organisms were identified to species level and antimicrobial susceptibility testing was performed manually by the disc diffusion technique or in the Vitek-2 (Biomereux, France) instrument. Interpretations were as per Clinical laboratory Standards Institute (CLSI) guidelines. Results: A total of 1150 bacterial isolates were cultured from 884 test samples during the study period. Of these 227 were Gram-positive and 923 were Gram-negative organisms. Staphylococcus aureus (99 isolates) was the commonest Gram-positive isolate followed by Enterococcus (79) and Gr A Streptococcus (30). Among the Gram negatives, E. coli (304), Pseudomonas aeruginosa (201) and Klebsiella pneumoniae (190) were the most common. Of the Staphylococcus aureus isolates 27.2% were methicillin resistant. Only 5.06% enterococci were vancomycin resistant. High rates of resistance to cefotaxime and ciprofloxacin were seen amongst E. coli (84.8% & 83.55%) and Klebsiella pneumoniae (71 & 62.1%) respectively. Resistance to carbapenems (meropenem) was high at 70% in Acinetobacter spp.; however all isolates were sensitive to colistin. Among the aminoglycosides, amikacin retained good efficacy against Escherichia coli (82.9%) and Pseudomonas aeruginosa (78.1%). Occasional isolates of emerging pathogens such as Chryseobacterium indologens, Roseomonas, and Achromobacter xyloxidans were also recovered. Conclusion: The common infections in cancer patients include respiratory, wound, tract infections and sepsis. The commonest isolates include Staphylococcus aureus, Enterococci, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa. There is a high level of resistance to the commonly used antibiotics among Gram-negative organisms.

Keywords: bacteria, resistance, infection, cancer

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67 The Effects of Grape Waste Bioactive Compounds on the Immune Response and Oxidative Stress in Pig Kidney

Authors: Mihai Palade, Gina Cecilia Pistol, Mariana Stancu, Veronica Chedea, Ionelia Taranu

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Nutrition is an important determinant of general health status, with especially focus on prevention and/or attenuation of the inflammatory-associated pathologies. People with chronic kidney disease can experience chronic inflammation that can lead to cardiovascular disease and even an increased rate of death. There are important links between chronic kidney diseases, inflammation and nutritional strategies that may prevent or protect against undesirable inflammation and oxidative stress. The grape by-products either seeds or pomace are rich in polyphenols which may be beneficial in prevention of inflammatory, antioxidant and antimicrobial processes. As a model for studying the impact of grape seeds on renal inflammation and oxidative stress, we used in this study weaned piglets. After a feeding trial of 30 days with a control diet and an experimental diet containing 5% grape seed (GS), kidney samples were collected. In renal tissues were determined the expression and activity of important markers of immune respose and oxidative stress: pro-inflammatory cytokines (TNF-alpha, IL-1 beta, IL-6, IL-8, IFN-gamma), anti-inflammatory cytokines (IL-4, IL-10), anti-oxidant enzymes (catalase CAT, superoxide dismutase SOD, glutathione peroxidise GPx) and important mediators belonging to nuclear receptors (NF-kB1, Nrf-2 and PPAR-gamma). Gene expression was evaluated by qPCR, whereas protein concentration was determined using proteomic techniques (ELISA). The activity of anti-oxidant enzymes was determined using specific kits. Our results showed that GS enriched in polyphenols does not have effect on TNF-alpha, IL-6 and IL-1 beta gene expression and protein concentration in kidney. By contrast, the gene expression and protein level of IL-8 and IFN-gamma were decreased in GS kidney. Anti-inflammatory cytokines IL-4 and IL-10 gene levels were increased in kidneys collected from GS piglets in comparison with controls, with no modification of protein levels between the two groups. The activities of anti-oxidant enzymes CAT and GPx were increased in kidney by GS, whereas SOD activity was unmodified in comparison with control samples. Also, the GS diet was associated with no modulation of mRNAs for nuclear receptors NF-kB1, Nrf-2 and PPAR-gamma gene expressions in kidneys. In conclusion, our results demonstrated that GS enriched in bioactive compounds such polyphenols could modulate inflammation and oxidative stress markers in kidney tissues. Further studies are necessary to elucidate the mechanism of action of GS compounds in case kidney inflammation associated with oxidative stress, and signalling molecules involved in these mechanisms.

Keywords: animal model, kidney inflammation, oxidative stress, grape seed

Procedia PDF Downloads 274
66 Control of an Outbreak of Vancomycin-Resistant Enterococci in a Tunisian Teaching Hospital

Authors: Hela Ghali, Sihem Ben Fredj, Mohamed Ben Rejeb, Sawssen Layouni, Salwa Khefacha, Lamine Dhidah, Houyem Said Laatiri

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Background: Antimicrobial resistance is a growing threat to public health and motivates to improve prevention and control programs both at international (WHO) and national levels. Despite their low pathogenicity, vancomycin-resistant enterococci (VRE) are common nosocomial pathogens in several countries. The high potential for transmission of VRE between patients and the threat to send its resistance genes to other bacteria such as staphylococcus aureus already resistant to meticilin, justify strict control measures. Indeed, in Europe, the proportion of Enterococcus faecium responsible for invasive infections, varies from 1% to 35% in 2011 and less than 5% were resistant to vancomycin. In addition, it represents the second cause of urinary tract and wound infections and the third cause of nosocomial bacteremia in the United States. The nosocomial outbreaks of VRE have been mainly described in intensive care services, hematology-oncology and haemodialysis. An epidemic of VRE has affected our hospital and the objective of this work is to describe the measures put in place. Materials/Methods: Following the alert given by the service of plastic surgery concerning a patient carrier of VRE, a team of the prevention and healthcare security service (doctor + technician) made an investigation. A review of files was conducted to draw the synoptic table and the table of cases. Results: By contacting the microbiology laboratory, we have identified four other cases of VRE and who were hospitalized in Medical resuscitation department (2 cases, one of them was transferred to the Physical rehabilitation department), and Nephrology department (2 cases). The visit has allowed to detect several malfunctions in professional practice. A crisis cell has allowed to validate, coordinate and implement control measures following the recommendations of the Technical Center of nosocomial infections. In fact, the process was to technically isolate cases in their sector of hospitalization, to restrict the use of antibiotics, to strength measures of basic hygiene, and to make a screening by rectal swab for both cases and contacts (other patients and health staff). These measures have helped to control the situation and no other case has been reported for a month. 2 new cases have been detected in the intensive care unit after a month. However, these are short-term strategies, and other measures in the medium and long term should be taken into account in order to face similar outbreaks. Conclusion: The efforts to control the outbreak were not efficient since 2 new cases have been reported after a month. Therefore, a continuous monitoring in order to detect new cases earlier is crucial to minimize the dissemination of VRE.

Keywords: hospitals, nosocomial infection, outbreak, vancomycin-resistant enterococci

Procedia PDF Downloads 277
65 Physico-Chemical and Biotechnological Characterization of Sheep’s Milk (Ovis aries) by Three Medicinal Plants Extracts

Authors: Fatima Bouazza, Khadija Khedid, Lamiae Amallah, Aziz Mouhaddach, Basma Boukour, Jihane Ennadir, Rachida Hassikou

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In order to combine milk and its derived products conservation and flavoring, Moroccans often used aromatic and medicinal plants. These plant extracts are endowed with several nutritive and therapeutic properties. This study constitutes a first national assessment of physico-chemical quality of sheep’s milk from moroccan Sardi breed and the evaluation of the antibacterial effect of three medicinal plants extracts: Aloe barbadensis Miller, Thymus satureioides and Mentha pulegium on flora isolated from this sheep's milk. 100 milk samples were collected in four regions of Morocco. The bacteria isolated were identified by classical and molecular methods (16S rRNA sequencing) and tested, according to the disk method, for their sensitivity to several antibiotics. The physico-chemical analyzes of sheep’s milk concerned the pH, titratable acidity, density, dry extract, freezing point and contents of: fat, proteins, lactose and calcium. The essential oils (EOs) of T. satureioides and M .pulegium were extracted by hydrodistillation and analyzed by GC / MS, while the Aloe vera leaf pulp was analyzed by the methods of Harborne and HPLC. A total number of 125 bacteria have been identified. Significant resistance to chemical antibiotics has been noted in LABs. The average temperature value of milk is around 57.15 °C, the pH is 6.56, the titratable acidity is around 3.4 ° D, the density is 1.035g / cm³ , the total dry extract is around 169.5g / l, the ash (9.8g / l), the freezing point (- 0.556 °C) while the average fat content is 67.85g / l . The samples richest in fat belong to the region of Settat, cradle of the Sardi breed, with a maximum average value of 74.4g / l. The average protein is 56g / l, lactose (39.92g / l), and calcium (1.855g / l). Analysis of the major components of EOs revealed the dominance of borneol in the case of T. satureioides and of pulegone in M. pulegium. Aloe vera gel contains alkaloids, flavonoids, catechic tannins, saponins and 1.60 µg / ml of aloin. The plant extracts have a bactericidal effect on E. coli, Klebsiellaoxytoca and Staphylococci and bacteriostatic effect on LABs of technological interest (Lactobacillus). As a result of this study, it is believed that the consumption of sardi sheep’s milk would be of nutritional benefit. Its richness in fat and proteins predisposes it for biotechnological development in the manufacture of cheese and yogurt. Also, the use of aromatic and medicinal plants, as natural additives would be of great benefit to flavor and maintain its quality.

Keywords: sheep’s milk, lactic flora, antimicrobial power, aloe barbadensis miller, thymus satureioides, mentha pulegium

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64 Antibacterial Effects of Zinc Oxide Nanoparticles as Alternative Therapy on Drug-Resistant Group B Streptococcus Strains Isolated from Pregnant Women

Authors: Leila Fozouni, Anahita Mazandarani

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Background: Maternal infections are the most common cause of infections in infants, and the level of infection and its severity highly depends on the degree of colonization of the bacteria in the mother; so, the occurrence of aggressive diseases is not unpredictable in mothers with very high colonization. Group B Streptococcus is part of the normal flora of the gastrointestinal and genital tracts in women and is the leading cause of septicemia and meningitis in newborns. Today Zinc oxide nanoparticle is regarded as one of the most commonly used and safest nanoparticles for defeating Gram-positive and Gram-negative bacteria. This study aims to determine the antibacterial effects of Zinc oxide on the growth of drug-resistant group B Streptococcus strains isolated from pregnant women. Materials and Methods: This cross-sectional study was conducted on 150 pregnant women of 28–37 weeks admitted to seven hospitals and maternity wards in Golestan province, northeast of Iran. For bacterial identification, rectovaginal swabs were firstly inoculated to the Todd-Hewitt Broth and cultured in blood agar (containing 5% sheep blood). Then microbiologic and PCR methods were performed to detect group B Streptococci. Disk diffusion and broth microdilution tests were used to determine the bacterial susceptibility to antibiotics according to CLSI M100(2021) criteria. The antibacterial properties of Zinc oxide nanoparticles were evaluated using the agar well-diffusion method. Results: The prevalence of group B Streptococcus was 18% in pregnant women. Out of twenty-seven positive cultures, 62.96% were higher than thirty years old. Ninety percent and 45% of isolates were resistant to clindamycin and erythromycin, respectively, and susceptibility to cefazolin was 71%. In addition, susceptibility to ampicillin and penicillin were 74% and 55%, respectively. The results showed that 82% of erythromycin-resistant, 92% clindamycin-resistant, and 78% of cefazolin-resistant isolates were eliminated by zinc oxide nanoparticles at a concentration of 100 mg/L of the nanoparticle. Furthermore, ZnONPs could inhibit all drug-resistant isolates at a concentration of 200 mg/mL (MIC90 ≥ 200). Conclusion: Since the drug resistance of group B streptococci against various antibiotics is increasing, determining and investigating the drug-resistance pattern of this bacterium to different antibiotics in order to prevent arbitrary consumption of antibiotics by pregnant women and ultimately prevent Infant mortality seems necessary. Generally, ZnONPs showed a high antimicrobial effect, and it was revealed that the bactericide effect increases upon the increase in the concentration of the nanoparticle.

Keywords: group B beta-hemolytic streptococcus, pregnant women, zinc oxide nanoparticles, drug resistance

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63 Biodegradable Self-Supporting Nanofiber Membranes Prepared by Centrifugal Spinning

Authors: Milos Beran, Josef Drahorad, Ondrej Vltavsky, Martin Fronek, Jiri Sova

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While most nanofibers are produced using electrospinning, this technique suffers from several drawbacks, such as the requirement for specialized equipment, high electrical potential, and electrically conductive targets. Consequently, recent years have seen the increasing emergence of novel strategies in generating nanofibers in a larger scale and higher throughput manner. The centrifugal spinning is simple, cheap and highly productive technology for nanofiber production. In principle, the drawing of solution filament into nanofibers using centrifugal spinning is achieved through the controlled manipulation of centrifugal force, viscoelasticity, and mass transfer characteristics of the spinning solutions. Engineering efforts of researches of the Food research institute Prague and the Czech Technical University in the field the centrifugal nozzleless spinning led to introduction of a pilot plant demonstrator NANOCENT. The main advantages of the demonstrator are lower investment cost - thanks to simpler construction compared to widely used electrospinning equipments, higher production speed, new application possibilities and easy maintenance. The centrifugal nozzleless spinning is especially suitable to produce submicron fibers from polymeric solutions in highly volatile solvents, such as chloroform, DCM, THF, or acetone. To date, submicron fibers have been prepared from PS, PUR and biodegradable polyesters, such as PHB, PLA, PCL, or PBS. The products are in form of 3D structures or nanofiber membranes. Unique self-supporting nanofiber membranes were prepared from the biodegradable polyesters in different mixtures. The nanofiber membranes have been tested for different applications. Filtration efficiencies for water solutions and aerosols in air were evaluated. Different active inserts were added to the solutions before the spinning process, such as inorganic nanoparticles, organic precursors of metal oxides, antimicrobial and wound healing compounds or photocatalytic phthalocyanines. Sintering can be subsequently carried out to remove the polymeric material and transfer the organic precursors to metal oxides, such as Si02, or photocatalytic Zn02 and Ti02, to obtain inorganic nanofibers. Electrospinning is more suitable technology to produce membranes for the filtration applications than the centrifugal nozzleless spinning, because of the formation of more homogenous nanofiber layers and fibers with smaller diameters. The self-supporting nanofiber membranes prepared from the biodegradable polyesters are especially suitable for medical applications, such as wound or burn healing dressings or tissue engineering scaffolds. This work was supported by the research grants TH03020466 of the Technology Agency of the Czech Republic.

Keywords: polymeric nanofibers, self-supporting nanofiber membranes, biodegradable polyesters, active inserts

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62 Use of Curcumin in Radiochemotherapy Induced Oral Mucositis Patients: A Control Trial Study

Authors: Shivayogi Charantimath

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Radiotherapy and chemotherapy are effective for treating malignancies but are associated with side effects like oral mucositis. Chlorhexidine gluconate is one of the most commonly used mouthwash in prevention of signs and symptoms of mucositis. Evidence shows that chlorhexidine gluconate has side effects in terms of colonization of bacteria, bad breadth and less healing properties. Thus, it is essential to find a suitable alternative therapy which is more effective with minimal side effects. Curcumin, an extract of turmeric is gradually being studied for its wide-ranging therapeutic properties such as antioxidant, analgesic, anti-inflammatory, antitumor, antimicrobial, antiseptic, chemo sensitizing and radio sensitizing properties. The present study was conducted to evaluate the efficacy and safety of topical curcumin gel on radio-chemotherapy induced oral mucositis in cancer patients. The aim of the study is to evaluate the efficacy and safety of curcumin gel in the management of oral mucositis in cancer patients undergoing radio chemotherapy and compare with chlorhexidine. The study was conducted in K.L.E. Society’s Belgaum cancer hospital. 40 oral cancer patients undergoing the radiochemotheraphy with oral mucositis was selected and randomly divided into two groups of 20 each. The study group A [20 patients] was advised Cure next gel for 2 weeks. The control group B [20 patients] was advised chlorhexidine gel for 2 weeks. The NRS, Oral Mucositis Assessment scale and WHO mucositis scale were used to determine the grading. The results obtained were calculated by using SPSS 20 software. The comparison of grading was done by applying Mann-Whitney U test and intergroup comparison was calculated by Wilcoxon matched pairs test. The NRS scores observed from baseline to 1st and 2nd week follow up in both the group showed significant difference. The percentage of change in erythema in respect to group A was 63.3% for first week and for second week, changes were 100.0% with p = 0.0003. The changes in Group A in respect to erythema was 34.6% for 1st week and 57.7% in second week. The intergroup comparison was significant with p value of 0.0048 and 0.0006 in relation to group A and group B respectively. The size of the ulcer score was measured which showed 35.5% [P=0.0010] of change in Group A for 1st and 2nd week showed totally reduction i.e. 103.4% [P=0.0001]. Group B showed 24.7% change from baseline to 1st week and 53.6% for 2nd week follow up. The intergroup comparison with Wilcoxon matched pair test was significant with p=0.0001 in group A. The result obtained by WHO mucositis score in respect to group A shows 29.6% [p=0.0004] change in first week and 75.0% [p=0.0180] change in second week which is highly significant in comparison to group B. Group B showed minimum changes i.e. 20.1% in 1st week and 33.3% in 2nd week. The p value with Wilcoxon was significant with 0.0025 in Group A for 1st week follow up and 0.000 for 2nd week follow up. Curcumin gel appears to an effective and safer alternative to chlorhexidine gel in treatment of oral mucositis.

Keywords: curcumin, chemotheraphy, mucositis, radiotheraphy

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61 The Prevalence and Profile of Extended Spectrum B-Lactamase (ESBL) Producing Enterobacteriaceae Species in the Intensive Care Unit (ICU) Setting of a Tertiary Care Hospital of North India

Authors: Harmeet Pal Singh Dhooria, Deepinder Chinna, UPS Sidhu, Alok Jain

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Serious infections caused by gram-negative bacteria are a significant cause of mortality and morbidity in the hospital setting. In acute care facilities like in intensive care units (ICUs), the intensity of antimicrobial use together with a population highly susceptible to infection, creates an environment, which facilitates both emergence and transmission of Extended Spectrum -lactamase (ESBL) producing Enterobacteriaceae species. The study was conducted in the Medical Intensive Care Unit (MICU) and the Pulmonary Critical Care Unit (PCCU) of the Department of Medicine, Dayanand Medical College and Hospital, Ludhiana, Punjab, India. Out of a total of 1108 samples of urine, blood and respiratory tract secretions received for culture and sensitivity analysis from Medical Intensive Care Unit and Pulmonary Critical Care Unit, a total of 170 isolates of Enterobacteriaceae species were obtained which were then included in our study. Out of these 170 isolates, confirmed ESBL production was seen in 116 (68.24%) cases. E.coli was the most common species isolated (56.47%) followed by Klebsiella (32.94%), Enterobacter (5.88%), Citrobacter (3.53%), Enterobacter (0.59%) and Morganella (0.59%) among the total isolates. The rate of ESBL production was more in Klebsiella (78.57%) as compared to E.coli (60.42%). ESBL producers were found to be significantly more common in patients with prior history of hospitalization, antibiotic use, and prolonged ICU stay. Also significantly increased the prevalence of ESBL related infections was observed in patients with a history of catheterization or central line insertion but not in patients with the history of intubation. Patients who had an underlying malignancy had significantly higher prevalence of ESBL related infections as compared to other co-morbid illnesses. A slightly significant difference in the rate of mortality/LAMA was observed in the ESBL producer versus the non-ESBL producer group. The rate of mortality/LAMA was significantly higher in the ESBL related UTI but not in the ESBL related respiratory tract and bloodstream infections. ESBL producing isolates had significantly higher rates of resistance to Cefepime and Piperacillin/Tazobactum, and to non β-lactum antibiotics like Amikacin and Ciprofloxacin. The level of resistance to Imipenem was lower as compared to other antibiotics. However, it was noted that ESBL producing isolates had higher levels of resistance to Imipenem as compared to non-ESBL producing isolates. Conclusion- The prevalence of ESBL producing organisms was found to be very high (68.24%) among Enterobacteriaceae isolates in our ICU setting as among other ICU care settings around the world.

Keywords: enterobacteriaceae, extended spectrum B-lactamase (ESBL), ICU, antibiotic resistance

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60 A Case of Prosthetic Vascular-Graft Infection Due to Mycobacterium fortuitum

Authors: Takaaki Nemoto

Abstract:

Case presentation: A 69-year-old Japanese man presented with a low-grade fever and fatigue that had persisted for one month. The patient had an aortic dissection on the aortic arch 13 years prior, an abdominal aortic aneurysm seven years prior, and an aortic dissection on the distal aortic arch one year prior, which were all treated with artificial blood-vessel replacement surgery. Laboratory tests revealed an inflammatory response (CRP 7.61 mg/dl), high serum creatinine (Cr 1.4 mg/dL), and elevated transaminase (AST 47 IU/L, ALT 45 IU/L). The patient was admitted to our hospital on suspicion of prosthetic vascular graft infection. Following further workups on the inflammatory response, an enhanced chest computed tomography (CT) and a non-enhanced chest DWI (MRI) were performed. The patient was diagnosed with a pulmonary fistula and a prosthetic vascular graft infection on the distal aortic arch. After admission, the patient was administered Ceftriaxion and Vancomycine for 10 days, but his fever and inflammatory response did not improve. On day 13 of hospitalization, a lung fistula repair surgery and an omental filling operation were performed, and Meropenem and Vancomycine were administered. The fever and inflammatory response continued, and therefore we took repeated blood cultures. M. fortuitum was detected in a blood culture on day 16 of hospitalization. As a result, we changed the treatment regimen to Amikacin (400 mg/day), Meropenem (2 g/day), and Cefmetazole (4 g/day), and the fever and inflammatory response began to decrease gradually. We performed a test of sensitivity for Mycobacterium fortuitum, and found that the MIC was low for fluoroquinolone antibacterial agent. The clinical course was good, and the patient was discharged after a total of 8 weeks of intravenous drug administration. At discharge, we changed the treatment regimen to Levofloxacin (500 mg/day) and Clarithromycin (800 mg/day), and prescribed these two drugs as a long life suppressive therapy. Discussion: There are few cases of prosthetic vascular graft infection caused by mycobacteria, and a standard therapy remains to be established. For prosthetic vascular graft infections, it is ideal to provide surgical and medical treatment in parallel, but in this case, surgical treatment was difficult and, therefore, a conservative treatment was chosen. We attempted to increase the treatment success rate of this refractory disease by conducting a susceptibility test for mycobacteria and treating with different combinations of antimicrobial agents, which was ultimately effective. With our treatment approach, a good clinical course was obtained and continues at the present stage. Conclusion: Although prosthetic vascular graft infection resulting from mycobacteria is a refractory infectious disease, it may be curative to administer appropriate antibiotics based on the susceptibility test in addition to surgical treatment.

Keywords: prosthetic vascular graft infection, lung fistula, Mycobacterium fortuitum, conservative treatment

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59 Prevalence of Antibiotic Resistant Enterococci in Treated Wastewater Effluent in Durban, South Africa and Characterization of Vancomycin and High-Level Gentamicin-Resistant Strains

Authors: S. H. Gasa, L. Singh, B. Pillay, A. O. Olaniran

Abstract:

Wastewater treatment plants (WWTPs) have been implicated as the leading reservoir for antibiotic resistant bacteria (ARB), including Enterococci spp. and antibiotic resistance genes (ARGs), worldwide. Enterococci are a group of clinically significant bacteria that have gained much attention as a result of their antibiotic resistance. They play a significant role as the principal cause of nosocomial infections and dissemination of antimicrobial resistance genes in the environment. The main objective of this study was to ascertain the role of WWTPs in Durban, South Africa as potential reservoirs for antibiotic resistant Enterococci (ARE) and their related ARGs. Furthermore, the antibiogram and resistance gene profile of Enterococci species recovered from treated wastewater effluent and receiving surface water in Durban were also investigated. Using membrane filtration technique, Enterococcus selective agar and selected antibiotics, ARE were enumerated in samples (influent, activated sludge, before chlorination and final effluent) collected from two WWTPs, as well as from upstream and downstream of the receiving surface water. Two hundred Enterococcus isolates recovered from the treated effluent and receiving surface water were identified by biochemical and PCR-based methods, and their antibiotic resistance profiles determined by the Kirby-Bauer disc diffusion assay, while PCR-based assays were used to detect the presence of resistance and virulence genes. High prevalence of ARE was obtained at both WWTPs, with values reaching a maximum of 40%. The influent and activated sludge samples contained the greatest prevalence of ARE with lower values observed in the before and after chlorination samples. Of the 44 vancomycin and high-level gentamicin-resistant isolates, 11 were identified as E. faecium, 18 as E. faecalis, 4 as E. hirae while 11 are classified as “other” Enterococci species. High-level aminoglycoside resistance for gentamicin (39%) and vancomycin (61%) was recorded in species tested. The most commonly detected virulence gene was the gelE (44%), followed by asa1 (40%), while cylA and esp were detected in only 2% of the isolates. The most prevalent aminoglycoside resistance genes were aac(6')-Ie-aph(2''), aph(3')-IIIa, and ant(6')-Ia detected in 43%, 45% and 41% of the isolates, respectively. Positive correlation was observed between resistant phenotypes to high levels of aminoglycosides and presence of all aminoglycoside resistance genes. Resistance genes for glycopeptide: vanB (37%) and vanC-1 (25%), and macrolide: ermB (11%) and ermC (54%) were detected in the isolates. These results show the need for more efficient wastewater treatment and disposal in order to prevent the release of virulent and antibiotic resistant Enterococci species and safeguard public health.

Keywords: antibiogram, enterococci, gentamicin, vancomycin, virulence signatures

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58 Development of Polylactic Acid Insert with a Cinnamaldehyde-Betacyclodextrin Complex for Cape Gooseberry (Physalis Peruviana L.) Packed

Authors: Gómez S. Jennifer, Méndez V. Camila, Moncayo M. Diana, Vega M. Lizeth

Abstract:

The cape gooseberry is a climacteric fruit; Colombia is one of the principal exporters in the world. The environmental condition of temperature and relative moisture decreases the titratable acidity and pH. These conditions and fruit maturation result in the fungal proliferation of Botrytis cinerea disease. Plastic packaging for fresh cape gooseberries was used for mechanical damage protection but created a suitable atmosphere for fungal growth. Beta-cyclodextrins are currently implemented as coatings for the encapsulation of hydrophobic compounds, for example, with bioactive compounds from essential oils such as cinnamaldehyde, which has a high antimicrobial capacity. However, it is a volatile substance. In this article, the casting method was used to obtain a polylactic acid (PLA) polymer film containing the beta-cyclodextrin-cinnamaldehyde inclusion complex, generating an insert that allowed the controlled release of the antifungal substance in packed cape gooseberries to decrease contamination by Botrytis cinerea in a latent state during storage. For the encapsulation technique, three ratios for the cinnamaldehyde: beta-cyclodextrin inclusion complex were proposed: (25:75), (40:60), and (50:50). Spectrophotometry, colorimetry in L*a*b* coordinate space and scanning electron microscopy (SEM) were made for the complex characterization. Subsequently, two ratios of tween and water (40:60) and (50:50) were used to obtain the polylactic acid (PLA) film. To determine mechanical and physical parameters of colourimetry in L*a*b* coordinate space, atomic force microscopy and stereoscopy were done to determine the transparency and flexibility of the film; for both cases, Statgraphics software was used to determine the best ratio in each of the proposed phases, where for encapsulation it was (50:50) with an encapsulation efficiency of 65,92%, and for casting the ratio (40:60) obtained greater transparency and flexibility that permitted its incorporation into the polymeric packaging. A liberation assay was also developed under ambient temperature conditions to evaluate the concentration of cinnamaldehyde inside the packaging through gas chromatography for three weeks. It was found that the insert had a controlled release. Nevertheless, a higher cinnamaldehyde concentration is needed to obtain the minimum inhibitory concentration for the fungus Botrytis cinerea (0.2g/L). The homogeneity of the cinnamaldehyde gas phase inside the packaging can be improved by considering other insert configurations. This development aims to impact emerging food preservation technologies with the controlled release of antifungals to reduce the affectation of the physico-chemical and sensory properties of the fruit as a result of contamination by microorganisms in the postharvest stage.

Keywords: antifungal, casting, encapsulation, postharvest

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57 Sequential Pulsed Electric Field and Ultrasound Assisted Extraction of Bioactive Enriched Fractions from Button Mushroom Stalks

Authors: Bibha Kumari, Nigel P. Brunton, Dilip K. Rai, Brijesh K. Tiwari

Abstract:

Edible mushrooms possess numerous functional components like homo- and hetero- β-glucans [β(1→3), β(1→4) and β(1→6) glucosidic linkages], chitins, ergosterols, bioactive polysaccharides and peptides imparting health beneficial properties to mushrooms. Some of the proven biological activities of mushroom extracts are antioxidant, antimicrobial, immunomodulatory, cholesterol lowering activity by inhibiting a key cholesterol metabolism enzyme i.e. 3-hydroxy-3-methyl-glutaryl CoA reductase (HMGCR), angiotensin I-converting enzyme (ACE) inhibition. Application of novel extraction technologies like pulsed electric field (PEF) and high power ultrasound offers clean, green, faster and efficient extraction alternatives with enhanced and good quality extracts. Sequential PEF followed by ultrasound assisted extraction (UAE) were applied to recover bioactive enriched fractions from industrial white button mushroom (Agaricus bisporus) stalk waste using environmentally friendly and GRAS solvents i.e. water and water/ethanol combinations. The PEF treatment was carried out at 60% output voltage, 2 Hz frequency for 500 pulses of 20 microseconds pulse width, using KCl salt solution of 0.6 mS/cm conductivity by the placing 35g of chopped fresh mushroom stalks and 25g of salt solution in the 4x4x4cm3 treatment chamber. Sequential UAE was carried out on the PEF pre-treated samples using ultrasonic-water-bath (USB) of three frequencies (25 KHz, 35 KHz and 45 KHz) for various treatment times (15-120 min) at 80°C. Individual treatment using either PEF or UAE were also investigation to compare the effect of each treatment along with the combined effect on the recovery and bioactivity of the crude extracts. The freeze dried mushroom stalk powder was characterised for proximate compositional parameters (dry weight basis) showing 64.11% total carbohydrate, 19.12% total protein, 7.21% total fat, 31.2% total dietary fiber, 7.9% chitin (as glucosamine equivalent) and 1.02% β-glucan content. The total phenolic contents (TPC) were determined by the Folin-Ciocalteu procedure and expressed as gallic-acid-equivalents (GAE). The antioxidant properties were ascertained using DPPH and FRAP assays and expressed as trolox-equivalents (TE). HMGCR activity and molecular mass of β-glucans will be measured using the commercial HMG-CoA Reductase Assay kit (Sigma-Aldrich) and size exclusion chromatography (HPLC-SEC), respectively. Effects of PEF, UAE and their combination on the antioxidant capacity, HMGCR inhibition and β-glucans content will be presented.

Keywords: β-glucan, mushroom stalks, pulsed electric field (PEF), ultrasound assisted extraction (UAE)

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56 Spectrum of Bacteria Causing Oral and Maxillofacial Infections and Their Antibiotic Susceptibility among Patients Attending Muhimbili National Hospital

Authors: Sima E. Rugarabamu, Mecky I. Matee, Elison N. M. Simon

Abstract:

Background: In Tanzania bacteriological studies of etiological agents of oro-facial infections are very limited, and very few have investigated anaerobes. The aim of this study was to determine the spectrum of bacterial agents involved in oral and maxillofacial infections in patients attending Muhimbili National Hospital, Dar-es-salaam, Tanzania. Method: This was a hospital based descriptive cross-sectional study that was conducted in the Department of Oral and Maxillofacial Surgery of the Muhimbili National Hospital in Dar es Salaam, Tanzania from 1st January 2014 to 31st August 2014. Seventy (70) patients with various forms of oral and maxillofacial infections who were recruited for the study. The study participants were interviewed using a prepared questionnaire after getting their consent. Pus aspirate was cultured on Blood agar, Chocolate Agar, MacConkey agar and incubated aerobically at 37°C. Imported blood agar was used for anaerobic culture whereby they were incubated at 37°Cin anaerobic jars in an atmosphere of generated using commercial gas-generating kits in accordance with manufacturer’s instructions. Plates were incubated at 37°C for 24 hours (For aerobic culture and 48 hours for anaerobic cultures). Gram negative rods were identified using API 20E while all other isolates were identified by conventional biochemical tests. Antibiotic sensitivity testing for isolated aerobic and anaerobic bacteria was detected by the disk diffusion, agar dilution and E-test using routine and commercially available antibiotics used to treat oral facial infections. Results: This study comprised of 41 (58.5%) males and 29 (41.5%) females with a mean age of 32 years SD +/-15.1 and a range of 19 to 70 years. A total of 161 bacteria strains were isolated from specimens obtained from 70 patients which were an average of 2.3 isolates per patient. Of these 103 were aerobic organism and 58 were strict anaerobes. A complex mix of strict anaerobes and facultative anaerobes accounted for 87% of all infections.The most frequent aerobes isolated was streptococcus spp 70 (70%) followed by Staphylococcus spp 18 (18%). Other organisms such as Klebsiella spp 4 (4%), Proteus spp 5 (5%) and Pseudomonas spp 2 (2%) were also seen. The anaerobic group was dominated by Prevotella spp 25 (43%) followed by Peptostreptococcus spp 18 (31%); other isolates were Pseudomonas spp 2 (1%), black pigmented Pophyromonas spp 4 (5%), Fusobacterium spp 3 (3%) and Bacteroides spp 5 (8%). Majority of these organisms were sensitive to Amoxicillin (98%), Gentamycin (89%), and Ciprofloxacin (100%). A 40% resistance to metronidazole was observed in Bacteroides spp otherwise this drug and others displayed good activity against anaerobes. Conclusions: Oral and maxillofacial facial infections at Muhimbili National Hospital are mostly caused by streptococcus spp and Prevotella spp. Strict anaerobes accounted for 36% of all isolates. The profile of isolates should assist in selecting empiric therapy for infections of the oral and maxillofacial region. Inclusion of antimicrobial agents against anaerobic bacteria is highly recommended.

Keywords: bacteria, oral and maxillofacial infections, antibiotic susceptibility, Tanzania

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55 Factors Mitigating against the Use of Alternative to Antibiotics (Phytobiotics) In Poultry Production among Farming Households in Nigeria

Authors: Akinola Helen Olufunke, Soetan Olatunbosun Jonathan, Adeleye Oludamola

Abstract:

Introduction: Antibiotic resistance has grown significantly, which is a major cause for concern. There have not been many significant developments in antibiotics over the past few decades, and practically all of the ones that are currently in use are losing effectiveness against pathogenic germs. Researchers are starting to focus more on the physiologically active compounds found in plants, particularly phytobiotics in poultry production. Consumption of chicken products is among the greatest in the country, but numerous nations, including Nigeria, use excessive amounts of necessary antibiotics in poultry farming, endangering the safety of such goods (through antimicrobial residues). Drug resistance has become a widespread issue as a result of the risky use of antibiotics in the chicken production industry. In order to replace antibiotics, biotic or natural products like phytobiotics (also known as botanicals or phytogenics) have drawn a lot of interest. Phytobiotics or their components are thought to be a relatively recent category of natural herbs that have acquired acceptance and favor among chicken farmers. The addition of several phytobiotic additions to poultry feed has demonstrated its capacity to improve both the broiler and layer populations' productivity. Design: Experimental research design and cross-sectional study was carried out at every 300 purposively selected farming household in the six-geopolitical zone in Nigeria. Data Analysis: A semi-structured questionnaire was administered to each farmer, and quantitative data were analyzed using Statistical Package for Social Science (SPSS) while the Chi-square test was used to analyze factors mitigating the use of Phytobiotics. Result: The result shows that the benefits associated with the use of phytobiotics are contributed to growth promotion in chickens and enhancement of productive performance of broiler and layer, which could be attributed to their antioxidant activity. The result further revealed that factors mitigating the use of phytobiotics were lack of knowledge in the use of phytobiotics, overdose or underdose usage, and seasonal availability of the phytobiotics. Others are the educational level of the farmers, intrinsic motivation, income poultry farming experience, price of phytobiotics based additives feeds, and intensity of extension agents in visiting them. Conclusion: The difficulties associated with using phytobiotics in chicken farms limit their willingness to boost productivity. The study found that most farmers were ignorant, which prevented them from handling this notion and turning their poultry into a viable enterprise while also allowing them to be creative. They believed that packing phytobiotics-based additive feed was expensive, and lastly, the seasonal availability of some phytobiotics. Recommendation: Further research in phytobiotics use in Nigeria should be carried out in order to establish its efficiency, safety, and awareness.

Keywords: mitigating, antibiotics, phytobiotics, poultry farming

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