Search results for: bacterial leaf spot
Commenced in January 2007
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Edition: International
Paper Count: 2032

Search results for: bacterial leaf spot

142 Biocontrol Potential of Trichoderma longibrachiatum as an Entomopathogenic Fungi against Bemisia tabaci

Authors: Waheed Anwar, Kiran Nawaz, Muhammad Saleem Haider, Ahmad Ali Shahid, Sehrish Iftikhar

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The whitefly, Bemisia tabaci (Gennadius), is a complex insect species, including many cryptic species or biotypes. Whitefly causes damage to many ornamental and horticultural crops through directly feeding on phloem sap, resulting in sooty mould and critically decreases the rate of photosynthesis of many host plants. Biological control has emerged as one of the most important methods for the management of soil-borne plant pathogens. Among the natural enemies of insects different entomopathogenic fungi are mostly used as biological control of the pest. The purpose of this research was to find indigenous insect-associated fungi and their virulence against Bemisia tabaci. A detailed survey of cotton fields in sample collection was conducted during July and August 2013 from the central mixed zone of Punjab, Pakistan. For the isolation of T. longibrachiatum, sabouraud dextrose peptone yeast extract agar (SDAY) media was used and morphological characterization of isolated T. longibrachiatum was studied using different dichotomous keys. Molecular Identification of the pathogen was confirmed by amplifying the internal transcribed spacer region. Blastn analysis showed 100% homology with already reported sequences on the database. For these bioassays, two conidial concentrations 4 × 108/mL & 4 × 104/mL of T. longibrachiatum was sprayed in clip cages for nymph and adult B. tabaci respectively under controlled environmental conditions. The pathogenicity of T. longibrachiatum was tested on nymph and adult whitefly to check mortality. Mortality of B. tabaci at nymphal and adult stages were observed after 24-hour intervals. Percentage mortality of nymphs treated with 4 x 104/mL conidia of T. longibrachiatum was 20, 24, 36 and 40% after 48, 72, 96, 72, 96, 120 and 144 hours respectively. However, no considerable difference was recorded in percentage mortality of whitefly after 120 and 144 hours. There were great variations after 24, 48, 72 and 96 hours in the rate of mortality. The efficacy of T. longibrachiatum as entomopathogenic fungi was evaluated in adult and nymphal stages of whitefly. Trichoderma longibrachiatum showed maximum activity on nymphal stages of whitefly as compared to adult stages. The percentage of conidial germination was also recorded on the outer surface of adult and nymphal stages of B. tabaci. The present findings indicated that T. longibrachiatum is an entomopathogenic fungus against B. tabaci and many species of Trichoderma were already reported as an antagonistc organism against a wide range of bacterial and fungal pathogens.

Keywords: efficacy, Trichoderma, virulence, bioassay

Procedia PDF Downloads 241
141 Investigations into the in situ Enterococcus faecalis Biofilm Removal Efficacies of Passive and Active Sodium Hypochlorite Irrigant Delivered into Lateral Canal of a Simulated Root Canal Model

Authors: Saifalarab A. Mohmmed, Morgana E. Vianna, Jonathan C. Knowles

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The issue of apical periodontitis has received considerable critical attention. Bacteria is integrated into communities, attached to surfaces and consequently form biofilm. The biofilm structure provides bacteria with a series protection skills against, antimicrobial agents and enhances pathogenicity (e.g. apical periodontitis). Sodium hypochlorite (NaOCl) has become the irrigant of choice for elimination of bacteria from the root canal system based on its antimicrobial findings. The aim of the study was to investigate the effect of different agitation techniques on the efficacy of 2.5% NaOCl to eliminate the biofilm from the surface of the lateral canal using the residual biofilm, and removal rate of biofilm as outcome measures. The effect of canal complexity (lateral canal) on the efficacy of the irrigation procedure was also assessed. Forty root canal models (n = 10 per group) were manufactured using 3D printing and resin materials. Each model consisted of two halves of an 18 mm length root canal with apical size 30 and taper 0.06, and a lateral canal of 3 mm length, 0.3 mm diameter located at 3 mm from the apical terminus. E. faecalis biofilms were grown on the apical 3 mm and lateral canal of the models for 10 days in Brain Heart Infusion broth. Biofilms were stained using crystal violet for visualisation. The model halves were reassembled, attached to an apparatus and tested under a fluorescence microscope. Syringe and needle irrigation protocol was performed using 9 mL of 2.5% NaOCl irrigant for 60 seconds. The irrigant was either left stagnant in the canal or activated for 30 seconds using manual (gutta-percha), sonic and ultrasonic methods. Images were then captured every second using an external camera. The percentages of residual biofilm were measured using image analysis software. The data were analysed using generalised linear mixed models. The greatest removal was associated with the ultrasonic group (66.76%) followed by sonic (45.49%), manual (43.97%), and passive irrigation group (control) (38.67%) respectively. No marked reduction in the efficiency of NaOCl to remove biofilm was found between the simple and complex anatomy models (p = 0.098). The removal efficacy of NaOCl on the biofilm was limited to the 1 mm level of the lateral canal. The agitation of NaOCl results in better penetration of the irrigant into the lateral canals. Ultrasonic agitation of NaOCl improved the removal of bacterial biofilm.

Keywords: 3D printing, biofilm, root canal irrigation, sodium hypochlorite

Procedia PDF Downloads 206
140 Combining Bio-Molecular and Isotopic Tools to Determine the Fate of Halogenated Compounds in Polluted Groundwater

Authors: N. Balaban, A. Buernstein, F. Gelman, Z. Ronen

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Brominated flame retardants are widespread pollutants, and are known to be toxic, carcinogenic, endocrinic disrupting as well as recalcitrant. The industrial complex Neot Hovav, in the Northern Negev, Israel, is situated above a fractured chalk aquitard, which is polluted by a wide variety of halogenated organic compounds. Two of the abundant pollutants found in the site are Dibromoneopentyl-glycol (DBNPG) and tribromoneopentyl-alcohol (TBNPA). Due to the elusive nature of the groundwater flow, it is difficult to connect between the spatial changes in contaminant concentrations to degradation. In this study, we attempt to determine whether these compounds are biodegraded in the groundwater, and to gain a better understanding concerning the bacterial community in the groundwater. This was achieved through the application of compound-specific isotope analysis (CSIA) of carbon (13^C/12^C) and bromine (81^Br/79^Br), and new-generation MiSeq pyrosequencing. The sampled boreholes were distributed among three main areas of the industrial complex: around the production plant of TBNPA and DBNPG; along the Hovav Wadi (small ephemeral stream) which crosses and drains the industrial complex; and downstream to the industrial area. TBNPA and DBNPG are found in all three areas, with no clear connection to the proximity of the borehole to the production plant. Initial isotopic data of TBNPA from boreholes in the area surrounding the production plant, reveal no changes in the carbon and bromine isotopic values. When observing the microbial groundwater community, the dominant phylum is Proteobacteria. Known anaerobic dehalogenating bacteria such as Dehalococcoides from the Chloroflexi phylum have also been detected. A statistical comparison of the groundwater microbial diversity using a multi-variant ordination of non-metric multidimensional scaling (NMDS) reveals three main clusters in accordance to spatial location in the industrial complex: all the boreholes sampled adjacent to the production plant cluster together and separately from the Wadi Hovav boreholes cluster and the downstream to the industrial area borehole cluster. This work provides the basis for the development and implication of an isotopic fractionation based tool for assessing the biodegradation of brominated organic compounds in contaminated environments, and a novel attempt to characterize the spatial microbial diversity in the contaminated site.

Keywords: biodegradation, brominated flame retardants, groundwater, isotopic fractionation, microbial diversity

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139 Degradation and Detoxification of Tetracycline by Sono-Fenton and Ozonation

Authors: Chikang Wang, Jhongjheng Jian, Poming Huang

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Among a wide variety of pharmaceutical compounds, tetracycline antibiotics are one of the largest groups of pharmaceutical compounds extensively used in human and veterinary medicine to treat and prevent bacterial infections. Because it is water soluble, biologically active, stable and bio-refractory, release to the environment threatens aquatic life and increases the risk posed by antibiotic-resistant pathogens. In practice, due to its antibacterial nature, tetracycline cannot be effectively destructed by traditional biological methods. Hence, in this study, two advanced oxidation processes such as ozonation and sono-Fenton processes were conducted individually to degrade the tetracycline for investigating their feasibility on tetracycline degradation. Effect of operational variables on tetracycline degradation, release of nitrogen and change of toxicity were also proposed. Initial tetracycline concentration was 50 mg/L. To evaluate the efficiency of tetracycline degradation by ozonation, the ozone gas was produced by an ozone generator (Model LAB2B, Ozonia) and introduced into the reactor with different flows (25 - 500 mL/min) at varying pH levels (pH 3 - pH 11) and reaction temperatures (15 - 55°C). In sono-Fenton system, an ultrasonic transducer (Microson VCX 750, USA) operated at 20 kHz combined with H₂O₂ (2 mM) and Fe²⁺ (0.2 mM) were carried out at different pH levels (pH 3 - pH 11), aeration gas and flows (air and oxygen; 0.2 - 1.0 L/min), tetracycline concentrations (10 - 200 mg/L), reaction temperatures (15 - 55°C) and ultrasonic powers (25 - 200 Watts), respectively. Sole ultrasound was ineffective on tetracycline degradation, where the degradation efficiencies were lower than 10% with 60 min reaction. Contribution of Fe²⁺ and H₂O₂ on the degradation of tetracycline was significant, where the maximum tetracycline degradation efficiency in sono-Fenton process was as high as 91.3% followed by 45.8% mineralization. Effect of initial pH level on tetracycline degradation was insignificant from pH 3 to pH 6 but significantly decreased as the pH was greater than pH 7. Increase of the ultrasonic power was slightly increased the degradation efficiency of tetracycline, which indicated that the hydroxyl radicals dominated the oxidation of tetracycline. Effects of aeration of air or oxygen with different flows and reaction temperatures were insignificant. Ozonation showed better efficiencies in tetracycline degradation, where the optimum reaction condition was found at pH 3, 100 mL O₃/min and 25°C with 94% degradation and 60% mineralization. The toxicity of tetracycline was significantly decreased due to the mineralization of tetracycline. In addition, less than 10% of nitrogen content was released to solution phase as NH₃-N, and the most degraded tetracycline cannot be full mineralized to CO₂. The results shown in this study indicated that both the sono-Fenton process and ozonation can effectively degrade the tetracycline and reduce its toxicity at profitable condition. The costs of two systems needed to be further investigated to understand the feasibility in tetracycline degradation.

Keywords: degradation, detoxification, mineralization, ozonation, sono-Fenton process, tetracycline

Procedia PDF Downloads 226
138 Synthesis, Physicochemical Characterization and Study of the Antimicrobial Activity of Chlorobutanol

Authors: N. Hadhoum, B. Guerfi, T. M. Sider, Z. Yassa, T. Djerboua, M. Boursouti, M. Mamou, F. Z. Hadjadj Aoul, L. R. Mekacher

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Introduction and objectives: Chlorobutanol is a raw material, mainly used as an antiseptic and antimicrobial preservative in injectable and ophthalmic preparations. The main objective of our study was the synthesis and evaluation of the antimicrobial activity of chlorobutanol hemihydrates. Material and methods: Chlorobutanol was synthesized according to the nucleophilic addition reaction of chloroform to acetone, identified by an infrared absorption using Spectrum One FTIR spectrometer, melting point, Scanning electron microscopy and colorimetric reactions. The dosage of carvedilol active substance was carried out by assaying the degradation products of chlorobutanol in a basic solution. The chlorobutanol obtained was subjected to bacteriological tests in order to study its antimicrobial activity. The antibacterial activity was evaluated against strains such as Escherichia coli (ATCC 25 922), Staphylococcus aureus (ATCC 25 923) and Pseudomonas aeroginosa (ATCC = American type culture collection). The antifungal activity was evaluated against human pathogenic fungal strains, such as Candida albicans and Aspergillus niger provided by the parasitology laboratory of the Hospital of Tizi-Ouzou, Algeria. Results and discussion: Chlorobutanol was obtained in an acceptable yield. The characterization tests of the product obtained showed a white and crystalline appearance (confirmed by scanning electron microscopy), solubilities (in water, ethanol and glycerol), and a melting temperature in accordance with the requirements of the European pharmacopoeia. The colorimetric reactions were directed towards the presence of a trihalogenated carbon and an alcohol function. The spectral identification (IR) showed the presence of characteristic chlorobutanol peaks and confirmed the structure of the latter. The microbiological study revealed an antimicrobial effect on all strains tested (Sataphylococcus aureus (MIC = 1250 µg/ml), E. coli (MIC = 1250 µg/ml), Pseudomonas aeroginosa (MIC = 1250 µg/ml), Candida albicans (MIC =2500 µg/ml), Aspergillus niger (MIC =2500 µg/ml)) with MIC values close to literature data. Conclusion: Thus, on the whole, the synthesized chlorobutanol satisfied the requirements of the European Pharmacopoeia, and possesses antibacterial and antifungal activity; nevertheless, it is necessary to insist on the purification step of the product in order to eliminate the maximum impurities.

Keywords: antimicrobial agent, bacterial and fungal strains, chlorobutanol, MIC, minimum inhibitory concentration

Procedia PDF Downloads 137
137 From the Perspective of a Veterinarian: The Future of Plant Raw Materials Used in the Feeding of Farm Animals

Authors: Ertuğrul Yılmaz

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One of the most important occupational groups in the food chain from farm to fork is a veterinary medicine. This occupational group, which has important duties in the prevention of many zoonotic diseases and in public health, takes place in many critical control points from soil to our kitchen. It has important duties from mycotoxins transmitted from the soil to slaughterhouses or milk processing facilities. Starting from the soil, which constitutes 70% of mycotoxin contamination, up to the TMR made from raw materials obtained from the soil, there are all critical control points from feeding to slaughterhouses and milk production enterprises. We can take the precaution of mycotoxins such as Aflatoxin B1, Ochratoxin, Zearalenone, and Fumonisin, which we encounter on farms while in the field. It has been reported that aflatoxin B1 is a casenerogen and passes into milk in studies. It is likely that many mycotoxins pose significant threats to public health and will turn out to be even more dangerous over time. Even raw material storage and TMR preparation are very important for public health. The danger of fumonisin accumulating in the liver will be understood over time. Zoonotic diseases are also explained with examples. In this study, how important veterinarians are in terms of public health is explained with examples. In the two-year mycotoxin screenings, fumonisin mycotoxin was found to be very high in corn and corn by-products, and it was determined that it accumulated in the liver for a long time and remained cornic in animals. It has been determined that mycotoxins are present in all livestock feeds, poultry feeds, and raw materials, not alone, but in double-triple form. Starting from the end, mycotoxin scans should be carried out from feed to raw materials and from raw materials to soil. In this way, we prevent the transmission of mycotoxins to animals and from animals to humans. Liver protectors such as toxin binders, beta-glucan, mannan oligosaccharides, activated carbon, prebiotics, and silymarin were used in certain proportions in the total mixed ratio, and positive results were obtained. Humidity and temperature controls of raw material silos were made at certain intervals. Necropsy was performed on animals that died as a result of mycotoxicosis, and macroscopic photographs were taken of the organs. We have determined that the mycotoxin screening in experimental animals and the feeds made without detecting the presence and amount of bacterial factors affect the results of the project to be made. For this, a series of precautionary plans have been created, starting from the production processes.

Keywords: mycotoxins, feed safety, processes, public health

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136 Surface Plasmon Resonance Imaging-Based Epigenetic Assay for Blood DNA Post-Traumatic Stress Disorder Biomarkers

Authors: Judy M. Obliosca, Olivia Vest, Sandra Poulos, Kelsi Smith, Tammy Ferguson, Abigail Powers Lott, Alicia K. Smith, Yang Xu, Christopher K. Tison

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Post-Traumatic Stress Disorder (PTSD) is a mental health problem that people may develop after experiencing traumatic events such as combat, natural disasters, and major emotional challenges. Tragically, the number of military personnel with PTSD correlates directly with the number of veterans who attempt suicide, with the highest rate in the Army. Research has shown epigenetic risks in those who are prone to several psychiatric dysfunctions, particularly PTSD. Once initiated in response to trauma, epigenetic alterations in particular, the DNA methylation in the form of 5-methylcytosine (5mC) alters chromatin structure and represses gene expression. Current methods to detect DNA methylation, such as bisulfite-based genomic sequencing techniques, are laborious and have massive analysis workflow while still having high error rates. A faster and simpler detection method of high sensitivity and precision would be useful in a clinical setting to confirm potential PTSD etiologies, prevent other psychiatric disorders, and improve military health. A nano-enhanced Surface Plasmon Resonance imaging (SPRi)-based assay that simultaneously detects site-specific 5mC base (termed as PTSD base) in methylated genes related to PTSD is being developed. The arrays on a sensing chip were first constructed for parallel detection of PTSD bases using synthetic and genomic DNA (gDNA) samples. For the gDNA sample extracted from the whole blood of a PTSD patient, the sample was first digested using specific restriction enzymes, and fragments were denatured to obtain single-stranded methylated target genes (ssDNA). The resulting mixture of ssDNA was then injected into the assay platform, where targets were captured by specific DNA aptamer probes previously immobilized on the surface of a sensing chip. The PTSD bases in targets were detected by anti-5-methylcytosine antibody (anti-5mC), and the resulting signals were then enhanced by the universal nanoenhancer. Preliminary results showed successful detection of a PTSD base in a gDNA sample. Brighter spot images and higher delta values (control-subtracted reflectivity signal) relative to those of the control were observed. We also implemented the in-house surface activation system for detection and developed SPRi disposable chips. Multiplexed PTSD base detection of target methylated genes in blood DNA from PTSD patients of severity conditions (asymptomatic and severe) was conducted. This diagnostic capability being developed is a platform technology, and upon successful implementation for PTSD, it could be reconfigured for the study of a wide variety of neurological disorders such as traumatic brain injury, Alzheimer’s disease, schizophrenia, and Huntington's disease and can be extended to the analyses of other sample matrices such as urine and saliva.

Keywords: epigenetic assay, DNA methylation, PTSD, whole blood, multiplexing

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135 Chemical Profiling of Hymenocardia acida Stem Bark Extract and Modulation of Selected Antioxidant and Esterase Enzymes in Kidney and Heart Ofwistar Rats

Authors: Adeleke G. E., Bello M. A., Abdulateef R. B., Olasinde T. T., Oriaje K. O., AransiI A., Elaigwu K. O., Omidoyin O. S., Shoyinka E. D., Awoyomi M. B., Akano M., Adaramoye O. A.

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Hymenocardia acidatul belongs to the genus, Hymenocardiaceae, which is widely distributed in Africa. Both the leaf and stem bark of the plant have been used in the treatment of several diseases. The present study examined the chemical constituents of the H. acida stem bark extract (HASBE) and its effects on some antioxidant indices and esterase enzymes in female Wistar rats. The HASBE was obtained by Soxhlet extraction using methanol and then subjected to Atomic Absorption Spectroscopy (AAS) for elemental analysis, and Fourier-Transform Infrared (FT-IR) spectroscopy, ultraviolet (UV) spectroscopy, for functional group analysis, while High-performance liquid chromatography (HPLC), and Gas Chromatography-Flame ionization detection (GC-FID) were carried out for compound identification. Forty-eight female Wistar rats were assigned into eight groups of six rats each and separately administered orally with normal saline (Control), 50, 100, 150, 200, 250, 300, 350 mg/kg of HASBE twice per week for eight weeks. The rats were sacrificed under chloroform anesthesia, and kidneys and heart were excised and processed to obtain homogenates. The levels of superoxide dismutase (SOD), catalase, Malondialdehyde (MDA), glutathione peroxidase (GPx), acetylcholinesterase (AChE), and carboxylesterase (CE) were determined spectrophotometrically. The AAS of HASBE shows the presence of eight elements, including Cobalt (0.303), Copper (0.222), Zinc (0.137), Iron (2.027), Nickel (1.304), Chromium (0.313), Manganese (0.213), and Magnesium (0.337 ppm). The FT-IR result of HASBE shows four peaks at 2961.4, 2926.0, 1056.7, and 1034.3 cm-1, while UV analysis shows a maximum absorbance (0.522) at 205 nm. The HPLC spectrum of HASBE indicates the presence of four major compounds, including orientin (77%), β-sitosterol (6.58%), rutin (5.02%), and betulinic acid (3.33%), while GC-FID result shows five major compounds, including rutin (53.27%), orientin (13.06%) and stigmasterol (11.73%), hymenocardine (6.43%) and homopterocarpin (5.29%). The SOD activity was significantly (p < 0.05) lowered in the kidney but elevated in the heart, while catalase was elevated in both organs relative to control rats. The GPx activity was significantly elevated only in the kidney, while MDA was not significantly (p > 0.05) affected in the two organs compared with controls. The activity of AChE was significantly elevated in both organs, while CE activity was elevated only in the kidney relative to control rats. The present study reveals that Hymenocardia acida stem bark extract majorly contains orientin, rutin, stigmasterol, hymenocardine, β-sitosterol, homopterocarpin, and betulinic acid. In addition, these compounds could possibly enhance redox status and esterase activities in the kidney and heart of Wistar rats.

Keywords: hymenocardia acida, elemental analysis, compounds identification, redox status, organs

Procedia PDF Downloads 119
134 Facilitating Primary Care Practitioners to Improve Outcomes for People With Oropharyngeal Dysphagia Living in the Community: An Ongoing Realist Review

Authors: Caroline Smith, Professor Debi Bhattacharya, Sion Scott

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Introduction: Oropharyngeal Dysphagia (OD) effects around 15% of older people, however it is often unrecognised and under diagnosed until they are hospitalised. There is a need for primary care healthcare practitioners (HCPs) to assume a proactive role in identifying and managing OD to prevent adverse outcomes such as aspiration pneumonia. Understanding the determinants of primary care HCPs undertaking this new behaviour provides the intervention targets for addressing. This realist review, underpinned by the Theoretical Domains Framework (TDF), aims to synthesise relevant literature and develop programme theories to understand what interventions work, how they work and under what circumstances to facilitate HCPs to prevent harm from OD. Combining realist methodology with behavioural science will permit conceptualisation of intervention components as theoretical behavioural constructs, thus informing the design of a future behaviour change intervention. Furthermore, through the TDF’s linkage to a taxonomy of behaviour change techniques, we will identify corresponding behaviour change techniques to include in this intervention. Methods & analysis: We are following the five steps for undertaking a realist review: 1) clarify the scope 2) Literature search 3) appraise and extract data 4) evidence synthesis 5) evaluation. We have searched Medline, Google scholar, PubMed, EMBASE, CINAHL, AMED, Scopus and PsycINFO databases. We are obtaining additional evidence through grey literature, snowball sampling, lateral searching and consulting the stakeholder group. Literature is being screened, evaluated and synthesised in Excel and Nvivo. We will appraise evidence in relation to its relevance and rigour. Data will be extracted and synthesised according to its relation to Initial programme theories (IPTs). IPTs were constructed after the preliminary literature search, informed by the TDF and with input from a stakeholder group of patient and public involvement advisors, general practitioners, speech and language therapists, geriatricians and pharmacists. We will follow the Realist and Meta-narrative Evidence Syntheses: Evolving Standards (RAMESES) quality and publication standards to report study results. Results: In this ongoing review our search has identified 1417 manuscripts with approximately 20% progressing to full text screening. We inductively generated 10 IPTs that hypothesise practitioners require: the knowledge to spot the signs and symptoms of OD; the skills to provide initial advice and support; and access to resources in their working environment to support them conducting these new behaviours. We mapped the 10 IPTs to 8 TDF domains and then generated a further 12 IPTs deductively using domain definitions to fulfil the remaining 6 TDF domains. Deductively generated IPTs broadened our thinking to consider domains such as ‘Emotion,’ ‘Optimism’ and ‘Social Influence’, e.g. If practitioners perceive that patients, carers and relatives expect initial advice and support, then they will be more likely to provide this, because they will feel obligated to do so. After prioritisation with stakeholders using a modified nominal group technique approach, a maximum of 10 IPTs will progress to test against the literature.

Keywords: behaviour change, deglutition disorders, primary healthcare, realist review

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133 Magnitude of Infection and Associated factor in Open Tibial Fractures Treated Operatively at Addis Ababa Burn Emergency and Trauma Center April, 2023

Authors: Tuji Mohammed Sani

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Back ground: An open tibial fracture is an injury where the fractured bone directly communicates with the outside environment. Due to the specific anatomical features of the tibia (limited soft tissue coverage), more than quarter of its fractures are classified as open, representing the most common open long-bone injuries. Open tibial fractures frequently cause significant bone comminution, periosteal stripping, soft tissue loss, contamination and are prone to bacterial entry with biofilm formation, which increases the risk of deep bone infection. Objective: The main objective of the study was to determine Prevalence of infection and its associated factors in surgically treated open tibial fracture in Addis Ababa Burn Emergency and Trauma (AaBET) center. Method: A facility based retrospective cross-sectional study was conducted among patient treated for open tibial fracture at AaBET center from September 2018 to September 2021. The data was collected from patient’s chart using structured data collection form, and Data was entered and analyzed using SPSS version 26. Bivariable and multiple binary logistic regression were fitted. Multicollinearity was checked among candidate variables using variance inflation factor and tolerance, which were less than 5 and greater than 0.2, respectively. Model adequacy were tested using Hosmer-Lemeshow goodness of fitness test (P=0.711). AOR at 95% CI was reported, and P-value < 0.05 was considered statistically significant. Result: This study found that 33.9% of the study participants had an infection. Initial IV antibiotic time (AOR=2.924, 95% CI:1.160- 7.370) and time of wound closure from injury (AOR=3.524, 95% CI: 1.798-6.908), injury to admission time (AOR=2.895, 95% CI: 1.402 – 5.977). and definitive fixation method (AOR=0.244, 95% CI: 0.113 – 0.4508) were the factors found to have a statistically significant association with the occurrence of infection. Conclusion: The rate of infection in open tibial fractures indicates that there is a need to improve the management of open tibial fracture treated at AaBET center. Time from injury to admission, time from injury to first debridement, wound closure time, and initial Intra Venous antibiotic time from the injury are an important factor that can be readily amended to improve the infection rate. Whether wound closed before seven days or not were more important factor associated with occurrences of infection.

Keywords: infection, open tibia, fracture, magnitude

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132 Xylanase Impact beyond Performance: A Prebiotic Approach in Laying Hens

Authors: Veerle Van Hoeck, Ingrid Somers, Dany Morisset

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Anti-nutritional factors such as non-starch polysaccharides (NSP) are present in viscous cereals used to feed poultry. Therefore, exogenous carbohydrases are commonly added to monogastric feed to degrade these NSP. Our hypothesis is that xylanase not only improves laying hen performance and digestibility but also induces a significant shift in microbial composition within the intestinal tract and, thereby, can cause a prebiotic effect. In this context, a better understanding of whether and how the chicken gut flora can be modulated by xylanase is needed. To do so, in the herein laying hen study, the effects of dietary supplementation of xylanase on performance, digestibility, and cecal microbiome were evaluated. A total of 96 HiSex laying hens was used in this experiment (3 diets and 16 replicates of 2 hens). Xylanase was added to the diets at concentrations of 0, 45,000 (15 g/t XygestTM HT) and 90,000 U/kg (30 g/t Xygest HT). The diets were based on wheat (~55 %), soybean, and sunflower meal. The lowest dosage, 45,000 U/kg, significantly increased average egg weight and improved feed efficiency compared to the control treatment (p < 0.05). Egg quality parameters were significantly improved in the experiment in response to the xylanase addition. For example, during the last 28 days of the trial, the 45,000 U/kg and the 90,000 U/kg treatments exhibited an increase in Haugh units and albumin heights (p < 0.05). Compared with the control, organic matter digestibility and N retention were drastically improved in the 45,000 U/kg treatment group, which implies better nutrient digestibility at this lowest recommended dosage compared to the control (p < 0.05). Furthermore, gross energy and crude fat digestibility were improved significantly for birds fed 90,000 U/kg group compared to the control. Importantly, 16S rRNA gene analysis revealed that xylanase at 45,000 U/kg dosages can exert a prebiotic effect. This conclusion was drawn based on studying the sequence variation in the 16S rRNA gene in order to characterize diverse microbial communities of the cecal content. A significant increase in beneficial bacteria (Lactobacilli spp and Enterococcus casseliflavus) was documented when adding 45,000 U/kg xylanase to the diet of laying hens. In conclusion, dietary supplementation of xylanase, even at the lowest dose of (45,000 U/kg), significantly improved laying hen performance and digestibility. Furthermore, it is generally accepted that a proper bacterial balance between the number of beneficial bacteria and pathogenic bacteria in the intestine is vital for the host. It seems that the xylanase enzyme is able to modulate the laying hen microbiome beneficially and thus exerts a prebiotic effect. This microbiome plasticity in response to the xylanase provides an attractive target for stimulating intestinal health.

Keywords: laying hen, prebiotic, XygestTM HT, xylanase

Procedia PDF Downloads 103
131 Lactobacillus rhamnosus GG Increases the Re-Epithelialization Rate of Model Wounds by Stimulating Keratinocyte Migration in Ex-Vivo

Authors: W. Mohammedsaeed, A. J. Mcbain, S. M. Cruickshank, C. A. O’Neill

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Many studies have demonstrated the importance of probiotics and their potential therapeutic effects within the gut. Recently, the possible therapeutic effects of probiotics in other tissues have also begun to be investigated. Comparatively few studies have evaluated the use of topical probiotics in relation to the skin. In this study, we have conducted preliminary investigations into whether a well-known probiotic, Lactobacillus rhamnosus GG (LGG), can increase the rate of re-epithelialization in a model wound. Full-thickness skin was obtained from individuals undergoing elective cosmetic surgery. This skin was wounded using excisional punch and cultured using a serum-free medium, either in the presence or absence of L. rhamnosus GG lysate. Histological staining of the sections was performed with Haematoxylin& Eosin E to quantify “epithelial tongue length”. This is the length of the new epithelial ‘tongue’ that grows and covers the exposed dermis at the inner wound edges. The length of the new epithelial ‘tongue’ was compared in untreated section and section treated with and L. rhamnosus GG made using108CFU/ml bacterial cells. L. rhamnosus GG lysate enhanced significantly the re-epithelialisation of treated wounds compared with that of untreated wounds (P=0.005, n=3). Tongue length, at day 1 was 7.55μm 0.15, at day 3 it was 18.5μm 0.25 and at day 7 was 22.9μm 0.35. These results can be compared with untreated cultures in which tongue length was 3.25μm 0.35, day 3 was 9.65μm 0.25 and day 7 was 13.5μm 0.15 post-wounding. In ex-vivo proliferation and migration cells were measured by determining the expression of nuclear proliferation marker Ki-67 and the expression of Phosphorylated cortactin respectively demonstrated that L. rhamnosus GG significantly increased NHEK proliferation and migration rates relative to controls. However, the dominant mechanism was migration because in ex-vivo skin treated with the L. rhamnosus GG up-regulated the gene expression of the chemokine receptor and ligands CXCR2 and CXCL2 comparing with controls (P=0.02, P=0.03 respectively, n=3). High levels of CXCL2/CXCL2 have already been implicated in multiple aspects of stimulation of wound healing through activation of keratinocyte migration. These data demonstrate that lysates from Lactobacillus rhamnosus GG increase re-epithelialization by stimulation of keratinocyte migration. The current study identifies the partial mechanism that contribute to stimulating the wound-healing process ex vivo in response to L. rhamnosus GG lysate is an increase in the production of CXCL2/ CXCR2 in ex vivo models. The use of probiotic lysates potentially offers new options to develop treatments that could improve wound healing.

Keywords: Lactobacillus rhamnosus GG, wounds, migration, lysate

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130 The Ductile Fracture of Armor Steel Targets Subjected to Ballistic Impact and Perforation: Calibration of Four Damage Criteria

Authors: Imen Asma Mbarek, Alexis Rusinek, Etienne Petit, Guy Sutter, Gautier List

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Over the past two decades, the automotive, aerospace and army industries have been paying an increasing attention to Finite Elements (FE) numerical simulations of the fracture process of their structures. Thanks to the numerical simulations, it is nowadays possible to analyze several problems involving costly and dangerous extreme loadings safely and at a reduced cost such as blast or ballistic impact problems. The present paper is concerned with ballistic impact and perforation problems involving ductile fracture of thin armor steel targets. The target fracture process depends usually on various parameters: the projectile nose shape, the target thickness and its mechanical properties as well as the impact conditions (friction, oblique/normal impact...). In this work, the investigations are concerned with the normal impact of a conical head-shaped projectile on thin armor steel targets. The main aim is to establish a comparative study of four fracture criteria that are commonly used in the fracture process simulations of structures subjected to extreme loadings such as ballistic impact and perforation. Usually, the damage initiation results from a complex physical process that occurs at the micromechanical scale. On a macro scale and according to the following fracture models, the variables on which the fracture depends are mainly the stress triaxiality ƞ, the strain rate, temperature T, and eventually the Lode angle parameter Ɵ. The four failure criteria are: the critical strain to failure model, the Johnson-Cook model, the Wierzbicki model and the Modified Hosford-Coulomb model MHC. Using the SEM, the observations of the fracture facies of tension specimen and of armor steel targets impacted at low and high incident velocities show that the fracture of the specimens is a ductile fracture. The failure mode of the targets is petalling with crack propagation and the fracture facies are covered with micro-cavities. The parameters of each ductile fracture model have been identified for three armor steels and the applicability of each criterion was evaluated using experimental investigations coupled to numerical simulations. Two loading paths were investigated in this study, under a wide range of strain rates. Namely, quasi-static and intermediate uniaxial tension and quasi-static and dynamic double shear testing allow covering various values of stress triaxiality ƞ and of the Lode angle parameter Ɵ. All experiments were conducted on three different armor steel specimen under quasi-static strain rates ranging from 10-4 to 10-1 1/s and at three different temperatures ranging from 297K to 500K, allowing drawing the influence of temperature on the fracture process. Intermediate tension testing was coupled to dynamic double shear experiments conducted on the Hopkinson tube device, allowing to spot the effect of high strain rate on the damage evolution and the crack propagation. The aforementioned fracture criteria are implemented into the FE code ABAQUS via VUMAT subroutine and they were coupled to suitable constitutive relations allow having reliable results of ballistic impact problems simulation. The calibration of the four damage criteria as well as a concise evaluation of the applicability of each criterion are detailed in this work.

Keywords: armor steels, ballistic impact, damage criteria, ductile fracture, SEM

Procedia PDF Downloads 288
129 Effects of Macro and Micro Nutrients on Growth and Yield Performances of Tomato (Lycopersicon esculentum MILL.)

Authors: K. M. S. Weerasinghe, A. H. K. Balasooriya, S. L. Ransingha, G. D. Krishantha, R. S. Brhakamanagae, L. C. Wijethilke

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Tomato (Lycopersicon esculentum Mill.) is a major horticultural crop with an estimated global production of over 120 million metric tons and ranks first as a processing crop. The average tomato productivity in Sri Lanka (11 metric tons/ha) is much lower than the world average (24 metric tons/ha).To meet the tomato demand for the increasing population the productivity has to be intensified through the agronomic-techniques. Nutrition is one of the main factors which govern the growth and yield of tomato and the main nutrient source soil affect the plant growth and quality of the produce. Continuous cropping, improper fertilizer usage etc., cause widespread nutrient deficiencies. Therefore synthetic fertilizers and organic manures were introduced to enhance plant growth and maximize the crop yields. In this study, effects of macro and micronutrient supplementations on improvement of growth and yield of tomato were investigated. Selected tomato variety is Maheshi and plants were grown in Regional Agricultural and Research Centre Makadura under the Department of Agriculture recommended (DOA) macro nutrients and various combination of Ontario recommended dosages of secondary and micro fertilizer supplementations. There were six treatments in this experiment and each treatment was replicated in three times and each replicate consisted of six plants. Other than the DOA recommendation, five combinations of Ontario recommended dosage of secondary and micronutrients for tomato were also used as treatments. The treatments were arranged in a Randomized Complete Block Design. All cultural practices were carried out according to the DOA recommendations. The mean data was subjected to the statistical analysis using SAS package and mean separation (Duncan’s Multiple Range test at 5% probability level) procedures. Secondary and micronutrients containing treatments significantly increased most of the growth parameters. Plant height, plant girth, number of leaves, leaf area index etc. Fruits harvested from pots amended with macro, secondary and micronutrients performed best in terms of total yield; yield quality; to pots amended with DOA recommended dosage of fertilizer for tomato. It could be due to the application of all essential macro and micro nutrients that rise in photosynthetic activity, efficient translocation and utilization of photosynthates causing rapid cell elongation and cell division in actively growing region of the plant leading to stimulation of growth and yield were caused. The experiment revealed and highlighted the requirements of essential macro, secondary and micro nutrient fertilizer supplementations for tomato farming. The study indicated that, macro and micro nutrient supplementation practices can influence growth and yield performances of tomato fruits and it is a promising approach to get potential tomato yields.

Keywords: macro and micronutrients, tomato, SAS package, photosynthates

Procedia PDF Downloads 423
128 Structural and Binding Studies of Peptidyl-tRNA Hydrolase from Pseudomonas aeruginosa Provide a Platform for the Structure Based Inhibitor Design against Peptidyl-tRNA Hydrolase

Authors: Sujata Sharma, Avinash Singh, Lovely Gautam, Pradeep Sharma, Mau Sinha, Asha Bhushan, Punit Kaur, Tej P. Singh

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Peptidyl-tRNA hydrolase (Pth) Pth is an essential bacterial enzyme that catalyzes the release of free tRNA and peptide moeities from peptidyl tRNAs during stalling of protein synthesis. In order to design inhibitors of Pth from Pseudomonas aeruginosa (PaPth), we have determined the structures of PaPth in its native state and in the bound states with two compounds, amino acylate-tRNA analogue (AAtA) and 5-azacytidine (AZAC). The peptidyl-tRNA hydrolase gene from Pseudomonas aeruginosa was amplified by Phusion High-Fidelity DNA Polymerase using forward and reverse primers, respectively. The E. coliBL21 (λDE3) strain was used for expression of the recombinant peptidyl-tRNA hydrolase from Pseudomonas aeruginosa. The protein was purified using a Ni-NTA superflow column. The crystallization experiments were carried out using hanging drop vapour diffusion method. The crystals diffracted to 1.50 Å resolution. The data were processed using HKL-2000. The polypeptide chain of PaPth consists of 194 amino acid residues from Met1 to Ala194. The centrally located β-structure is surrounded by α-helices from all sides except the side that has entrance to the substrate binding site. The structures of the complexes of PaPth with AAtA and AZAC showed the ligands bound to PaPth in the substrate binding cleft and interacted with protein atoms extensively. The residues that formed intermolecular hydrogen bonds with the atoms of AAtA included Asn12, His22, Asn70, Gly113, Asn116, Ser148, and Glu161 of the symmetry related molecule. The amino acids that were involved in hydrogen bonded interactions in case of AZAC included, His22, Gly113, Asn116, and Ser148. As indicated by fittings of two ligands and the number of interactions made by them with protein atoms, AAtA appears to be a more compatible with the structure of the substrate binding cleft. However, there is a further scope to achieve a better stacking than that of O-tyrosyl moiety because it is not still ideally stacked. These observations about the interactions between the protein and ligands have provided the information about the mode of binding of ligands, nature and number of interactions. This information may be useful for the design of tight inhibitors of Pth enzymes.

Keywords: peptidyl tRNA hydrolase, Acinetobacter baumannii, Pth enzymes, O-tyrosyl

Procedia PDF Downloads 385
127 Polyvinyl Alcohol Incorporated with Hibiscus Extract Microcapsules as Combined Active and Intelligent Composite Film for Meat Preservation: Antimicrobial, Antioxidant, and Physicochemical Investigations

Authors: Ahmed F. Ghanem, Marwa I. Wahba, Asmaa N. El-Dein, Mohamed A. EL-Raey, Ghada E. A. Awad

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Numerous attempts are being performed in order to formulate suitable packaging materials for the meat products. However, to the best of our knowledge, the incorporation of the free hibiscus extract or its microcapsules in the pure polyvinyl alcohol (PVA) matrix as packaging materials for the meats is seldom reported. Therefore, this study aims at the protection of the aqueous crude extract of the hibiscus flowers utilizing the spry drying encapsulation technique. Results of the Fourier transform infrared (FTIR), the scanning electron microscope (SEM), and the particle size analyzer confirmed the successful formation of the assembled capsules via strong interactions, the spherical rough microparticles, and the particle size of ~ 235 nm, respectively. Also, the obtained microcapsules enjoy higher thermal stability than the free extract. Then, the obtained spray-dried particles were incorporated into the casting solution of the pure PVA film with a concentration of 10 wt. %. The segregated free-standing composite films were investigated, compared to the neat matrix, with several characterization techniques such as FTIR, SEM, thermal gravimetric analysis (TGA), mechanical tester, contact angle, water vapor permeability, and oxygen transmission. The results demonstrated variations in the physicochemical properties of the PVA film after the inclusion of the free and the extract microcapsules. Moreover, biological studies emphasized the biocidal potential of the hybrid films against the microorganisms contaminating the meat. Specifically, the microcapsules imparted not only antimicrobial but also antioxidant activities to the PVA matrix. Application of the prepared films on the real meat samples displayed a low bacterial growth with a slight increase in the pH over the storage time which continued up to 10 days at 4 oC, as further evidence to the meat safety. Moreover, the colors of the films did not significantly changed except after 21 days indicating the spoilage of the meat samples. No doubt, the dual-functional of the prepared composite films pave the way towards combined active and smart food packaging applications. This would play a vital role in the food hygiene, including also the quality control and the assurance.

Keywords: PVA, hibiscus, extraction, encapsulation, active packaging, smart and intelligent packaging, meat spoilage

Procedia PDF Downloads 61
126 Metagenomic Assessment of the Effects of Genetically Modified Crops on Microbial Ecology and Physicochemical Properties of Soil

Authors: Falana Yetunde Olaitan, Ijah U. J. J, Solebo Shakirat O.

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Genetically modified crops are already phenomenally successful and are grown worldwide in more than eighteen countries on more than 67 million hectares. Nigeria, in October 2018, approved Bacillus thuringiensis (Bt) cotton and maize; therefore, the need to carry out environmental risk assessment studies. A total of 15 4L octagonal ceramic pots were filled with 4kg of soil and placed on the bench in 2 rows of 10 pots each and the 3rd row of 5 pots, 1st-row pots were used to plant GM cotton seeds, while the 2nd-row pots were used for non-GM cotton seeds and the 3rd row of 5 pots served as control, all in the screen house. Soil samples for metagenomic DNA extraction were collected at random and at the monthly interval after planting at a distance of 2mm from the plant’s root and at a depth of 10cm using a sterile spatula. Soil samples for physicochemical analysis were collected before planting and after harvesting the GM and non-GM crops as well as from the control soil. The DNA was extracted, quantified and sequenced; Sample 1A (DNA from GM cotton Soil at 1st interval) gave the lowest sequence read with 0.853M while sample 2B (DNA from GM cotton Soil at 2nd interval) gave the highest with 5.785M, others gave between 1.8M and 4.7M. The samples treatment were grouped into four, Group 1 (GM cotton soil from 1 to 3 intervals) had between 800,000 and 5,700,000 strains of microbes (SOM), Group 2 (non GM cotton soil from 1 to 3 intervals) had between 1,400,600 and 4,200,000 SOM, Group 3 (control soil) had between 900,000 and 3,600,000 SOM and Group 4 (initial soil) had between 3,700,000 and 4,000,000 SOM. The microbes observed were predominantly bacteria (including archaea), fungi, dark matter alongside protists and phages. The predominant bacterial groups were the Terrabacteria (Bacillus funiculus, Bacillus sp.), the Proteobacteria (Microvirga massiliensis, sphingomonas sp.) and the Archaea (Nitrososphaera sp.), while the fungi were Aspergillus fischeri and Fusarium falciforme. The comparative analysis between groups was done using JACCARD PERMANOVA beta diversity analysis at P-value not more than 0.76 and there was no significant pair found. The pH for initial, GM cotton, non-GM cotton and control soil were 6.28, 6.26, 7.25, 8.26 and the percentage moisture was 0.63, 0.78, 0.89 and 0.82, respectively, while the percentage Nitrogen was observed to be 17.79, 1.14, 1.10 and 0.56 respectively. Other parameters include, varying concentrations of Potassium (0.46, 1,284.47, 1,785.48, 1,252.83 mg/kg) and Phosphorus (18.76, 17.76, 16.87, 15.23 mg/kg) were recorded for the four treatments respectively. The soil consisted mainly of silt (32.09 to 34.66%) and clay (58.89 to 60.23%), reflecting the soil texture as silty – clay. The results were then tested with ANOVA at less than 0.05 P-value and no pair was found to be significant as well. The results suggest that the GM crops have no significant effect on microbial ecology and physicochemical properties of the soil and, in turn, no direct or indirect effects on human health.

Keywords: genetically modified crop, microbial ecology, physicochemical properties, metagenomics, DNA, soil

Procedia PDF Downloads 121
125 Development of a Bioprocess Technology for the Production of Vibrio midae, a Probiotic for Use in Abalone Aquaculture

Authors: Ghaneshree Moonsamy, Nodumo N. Zulu, Rajesh Lalloo, Suren Singh, Santosh O. Ramchuran

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The abalone industry of South Africa is under severe pressure due to illegal harvesting and poaching of this seafood delicacy. These abalones are harvested excessively; as a result, these animals do not have a chance to replace themselves in their habitats, ensuing in a drastic decrease in natural stocks of abalone. Abalone has an extremely slow growth rate and takes approximately four years to reach a size that is market acceptable; therefore, it was imperative to investigate methods to boost the overall growth rate and immunity of the animal. The University of Cape Town (UCT) began to research, which resulted in the isolation of two microorganisms, a yeast isolate Debaryomyces hansenii and a bacterial isolate Vibrio midae, from the gut of the abalone and characterised them for their probiotic abilities. This work resulted in an internationally competitive concept technology that was patented. The next stage of research was to develop a suitable bioprocess to enable commercial production. Numerous steps were taken to develop an efficient production process for V. midae, one of the isolates found by UCT. The initial stages of research involved the development of a stable and robust inoculum and the optimization of physiological growth parameters such as temperature and pH. A range of temperature and pH conditions were evaluated, and data obtained revealed an optimum growth temperature of 30ᵒC and a pH of 6.5. Once these critical growth parameters were established further media optimization studies were performed. Corn steep liquor (CSL) and high test molasses (HTM) were selected as suitable alternatives to more expensive, conventionally used growth medium additives. The optimization of CSL (6.4 g.l⁻¹) and HTM (24 g.l⁻¹) concentrations in the growth medium resulted in a 180% increase in cell concentration, a 5716-fold increase in cell productivity and a 97.2% decrease in the material cost of production in comparison to conventional growth conditions and parameters used at the onset of the study. In addition, a stable market-ready liquid probiotic product, encompassing the viable but not culturable (VBNC) state of Vibrio midae cells, was developed during the downstream processing aspect of the study. The demonstration of this technology at a full manufacturing scale has further enhanced the attractiveness and commercial feasibility of this production process.

Keywords: probiotics, abalone aquaculture, bioprocess technology, manufacturing scale technology development

Procedia PDF Downloads 126
124 Inverted Diameter-Limit Thinning: A Promising Alternative for Mixed Populus tremuloides Stands Management

Authors: Ablo Paul Igor Hounzandji, Benoit Lafleur, Annie DesRochers

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Introduction: Populus tremuloides [Michx] regenerates rapidly and abundantly by root suckering after harvest, creating stands with interconnected stems. Pre-commercial thinning can be used to concentrate growth on fewer stems to reach merchantability faster than un-thinned stands. However, conventional thinning methods are typically designed to reach even spacing between residual stems (1,100 stem ha⁻¹, evenly distributed), which can lead to treated stands consisting of weaker/smaller stems compared to the original stands. Considering the nature of P. tremuloides's regeneration, with large underground biomass of interconnected roots, aiming to keep the most vigorous and largest stems, regardless of their spatial distribution, inverted diameter-limit thinning could be more beneficial to post-thinning stand productivity because it would reduce the imbalance between roots and leaf area caused by thinning. Aims: This study aimed to compare stand and stem productivity of P. tremuloides stands thinned with a conventional thinning treatment (CT; 1,100 stem ha⁻¹, evenly distributed), two levels of inverted diameter-limit thinning (DL1 and DL2, keeping the largest 1100 or 2200 stems ha⁻¹, respectively, regardless of their spatial distribution) and a control unthinned treatment. Because DL treatments can create substantial or frequent gaps in the thinned stands, we also aimed to evaluate the potential of this treatment to recreate mixed conifer-broadleaf stands by fill-planting Picea glauca seedlings. Methods: Three replicate 21 year-old sucker-regenerated aspen stands were thinned in 2010 according to four treatments: CT, DL1, DL2, and un-thinned control. Picea glauca seedlings were underplanted in gaps created by the DL1 and DL2 treatments. Stand productivity per hectare, stem quality (diameter and height, volume stem⁻¹) and survival and height growth of fill-planted P. glauca seedlings were measured 8 year post-treatments. Results: Productivity, volume, diameter, and height were better in the treated stands (CT, DL1, and DL2) than in the un-thinned control. Productivity of CT and DL1 stands was similar 4.8 m³ ha⁻¹ year⁻¹. At the tree level, diameter and height of the trees in the DL1 treatment were 5% greater than those in the CT treatment. The average volume of trees in the DL1 treatment was 11% higher than the CT treatment. Survival after 8 years of fill planted P. glauca seedlings was 2% greater in the DL1 than in the DL2 treatment. DL1 treatment also produced taller seedlings (+20 cm). Discussion: Results showed that DL treatments were effective in producing post-thinned stands with larger stems without affecting stand productivity. In addition, we showed that these treatments were suitable to introduce slower growing conifer seedlings such as Picea glauca in order to re-create or maintain mixed stands despite the aggressive nature of P. tremuloides sucker regeneration.

Keywords: Aspen, inverted diameter-limit, mixed forest, populus tremuloides, silviculture, thinning

Procedia PDF Downloads 115
123 Impact of Fluoride Contamination on Soil and Water at North 24 Parganas, West Bengal, India

Authors: Rajkumar Ghosh

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Fluoride contamination is a growing concern in various regions across the globe, including North 24 Parganas in West Bengal, India. The presence of excessive fluoride in the environment can have detrimental effects on crops, soil quality, and water resources. This note aims to shed light on the implications of fluoride contamination and its impact on the agricultural sector in North 24 Parganas. The agricultural lands in North 24 Parganas have been significantly affected by fluoride contamination, leading to adverse consequences for crop production. Excessive fluoride uptake by plants can hinder their growth, reduce crop yields, and impact the quality of agricultural produce. Certain crops, such as paddy, vegetables, and fruits, are more susceptible to fluoride toxicity, resulting in stunted growth, leaf discoloration, and reduced nutritional value. Fluoride-contaminated water, often used for irrigation, contributes to the accumulation of fluoride in the soil. Over time, this can lead to soil degradation and reduced fertility. High fluoride levels can alter soil pH, disrupt the availability of essential nutrients, and impair microbial activity critical for nutrient cycling. Consequently, the overall health and productivity of the soil are compromised, making it increasingly challenging for farmers to sustain agricultural practices. Fluoride contamination in North 24 Parganas extends beyond the soil and affects water resources as well. The excess fluoride seeps into groundwater, making it unsafe for consumption. Long-term consumption of fluoride-contaminated water can lead to various health issues, including dental and skeletal fluorosis. These health concerns pose significant risks to the local population, especially those reliant on contaminated water sources for their daily needs. Addressing fluoride contamination requires concerted efforts from various stakeholders, including government authorities, researchers, and farmers. Implementing appropriate water treatment technologies, such as defluoridation units, can help reduce fluoride levels in drinking water sources. Additionally, promoting alternative irrigation methods and crop diversification strategies can aid in mitigating the impact of fluoride on agricultural productivity. Furthermore, creating awareness among farmers about the adverse effects of fluoride contamination and providing access to alternative water sources are crucial steps toward safeguarding the health of the community and sustaining agricultural activities in the region. Fluoride contamination poses significant challenges to crop production, soil health, and water resources in North 24 Parganas, West Bengal. It is imperative to prioritize efforts to address this issue effectively and implement appropriate measures to mitigate fluoride contamination. By adopting sustainable practices and promoting awareness, the community can work towards restoring the agricultural productivity, soil quality and ensuring access to safe drinking water in the region.

Keywords: fluoride contamination, drinking water, toxicity, soil health

Procedia PDF Downloads 64
122 Multicenter Baseline Survey to Outline Antimicrobial Prescribing Practices at Six Public Sectortertiary Care Hospitals in a Low Middle Income Country

Authors: N. Khursheed, M. Fatima, S. Jamal, A. Raza, S. Rattani, Q. Ahsan, A. Rasheed, M. Jawed

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Introduction: Antibiotics are among the commonly prescribed medicines to treat bacterial infections. Their misuse intensifies resistance, and overuse incurs heavy losses to the healthcare system in terms of increased treatment costs and enhanced disease burden. Studies show that 40% of empirically used antibiotics are irrationally utilized. The objective of this study was to evaluate prescribing pattern of antibiotics at six public sector tertiary care hospitals across Pakistan. Methods: A multicenter cross-sectional point prevalence survey (PPS) was conducted in selected wards of six public sector tertiary care hospitals in Pakistan as part of the Clinical Engagement program by Fleming Fund Country Grant Pakistan in collaboration with Indus Hospital & Health Network (IHHN) from February to March 2021, these included Jinnah Postgraduate Medical Center and Dr. Ruth K. M. Pfau Civil Hospital from Karachi, Sheikh Zayed Hospital Lahore, Nishtar Medical University Hospital Multan, Medical Teaching Institute Hayatabad Medical Complex Peshawar, and Provincial Headquarters Hospital Gilgit. WHO PPS methodology was used for data collection (Hospital, ward, and patient level data was collected). Data was entered into the open-source Kobo Collect application and was analyzed using SPSS (version 22.0). Findings: Medical records of 837 in-patients were surveyed, of which the prevalence of antibiotics use was 78.5%. The most commonly prescribed antimicrobial was Ceftriaxone (21.7%) which is categorized in the Watch group of WHO AWaRe Classification, followed by Metronidazole (17.3%), Cefoperazone/Sulbactam (8.4%), Co-Amoxiclav (6.3%) and Piperacillin/Tazobactam (5.9%). The antibiotics were prescribed largely for surgical prophylaxis (36.7%), followed by community-acquired infections (24.7%). One antibiotic was prescribed to 46.7%, two to 39.9%, and three or more to 12.5 %. Two of six (30%) hospitals had functional drug and therapeutic committees, three (50%) had infection prevention and control committees, and one facility had an antibiotic formulary. Conclusion: Findings demonstrate high consumption of broad-spectrum antimicrobials and emphasizes the importance of expanding the antimicrobial stewardship program. Mentoring clinical teams will help to rationalize antimicrobial use.

Keywords: antimicrobial resistance, antimicrobial stewardship, point prevalence survey, antibiotics

Procedia PDF Downloads 73
121 Chemical Modifications of Three Underutilized Vegetable Fibres for Improved Composite Value Addition and Dye Absorption Performance

Authors: Abayomi O. Adetuyi, Jamiu M. Jabar, Samuel O. Afolabi

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Vegetable fibres are classes of fibres of low density, biodegradable and non-abrasive that are largely abundant fibre materials with specific properties and mostly found/ obtained in plants on earth surface. They are classified into three categories, depending on the part of the plant from which they are gotten from namely: fruit, Blast and Leaf fibre. Ever since four/five millennium B.C, attention has been focussing on the commonest and highly utilized cotton fibre obtained from the fruit of cotton plants (Gossypium spp), for the production of cotton fabric used in every home today. The present study, therefore, focused on the ability of three underutilized vegetable (fruit) fibres namely: coir fiber (Eleas coniferus), palm kernel fiber and empty fruit bunch fiber (Elias guinensis) through chemical modifications for better composite value addition performance to polyurethane form and dye adsorption. These fibres were sourced from their parents’ plants, identified and cleansed with 2% hot detergent solution 1:100, rinsed in distilled water and oven-dried to constant weight, before been chemically modified through alkali bleaching, mercerization and acetylation. The alkali bleaching involves treating 0.5g of each fiber material with 100 mL of 2% H2O2 in 25 % NaOH solution with refluxing for 2 h. While that of mercerization and acetylation involves the use of 5% sodium hydroxide NaOH solution for 2 h and 10% acetic acid- acetic anhydride 1:1 (v/v) (CH3COOH) / (CH3CO)2O solution with conc. H2SO4 as catalyst for 1 h, respectively on the fibres. All were subsequently washed thoroughly with distilled water and oven dried at 105 0C for 1 h. These modified fibres were incorporated as composite into polyurethane form and used in dye adsorption study of indigo. The first two treatments led to fiber weight reduction, while the acidified acetic anhydride treatment gave the fibers weight increment. All the treated fibers were found to be of less hydrophilic nature, better mechanical properties, higher thermal stabilities as well as better adsorption surfaces/capacities than the untreated ones. These were confirmed by gravimetric analysis, Instron Universal Testing Machine, Thermogravimetric Analyser and the Scanning Electron Microscope (SEM) respectively. The fiber morphology of the modified fibers showed smoother surfaces than unmodified fibres.The empty fruit bunch fibre and the coconut coir fibre are better than the palm kernel fibres as reinforcers for composites or as adsorbents for waste-water treatment. Acetylation and alkaline bleaching treatment improve the potentials of the fibres more than mercerization treatment. Conclusively, vegetable fibres, especially empty fruit bunch fibre and the coconut coir fibre, which are cheap, abundant and underutilized, can replace the very costly powdered activated carbon in wastewater treatment and as reinforcer in foam.

Keywords: chemical modification, industrial application, value addition, vegetable fibre

Procedia PDF Downloads 293
120 Artificial Intelligence Models for Detecting Spatiotemporal Crop Water Stress in Automating Irrigation Scheduling: A Review

Authors: Elham Koohi, Silvio Jose Gumiere, Hossein Bonakdari, Saeid Homayouni

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Water used in agricultural crops can be managed by irrigation scheduling based on soil moisture levels and plant water stress thresholds. Automated irrigation scheduling limits crop physiological damage and yield reduction. Knowledge of crop water stress monitoring approaches can be effective in optimizing the use of agricultural water. Understanding the physiological mechanisms of crop responding and adapting to water deficit ensures sustainable agricultural management and food supply. This aim could be achieved by analyzing and diagnosing crop characteristics and their interlinkage with the surrounding environment. Assessments of plant functional types (e.g., leaf area and structure, tree height, rate of evapotranspiration, rate of photosynthesis), controlling changes, and irrigated areas mapping. Calculating thresholds of soil water content parameters, crop water use efficiency, and Nitrogen status make irrigation scheduling decisions more accurate by preventing water limitations between irrigations. Combining Remote Sensing (RS), the Internet of Things (IoT), Artificial Intelligence (AI), and Machine Learning Algorithms (MLAs) can improve measurement accuracies and automate irrigation scheduling. This paper is a review structured by surveying about 100 recent research studies to analyze varied approaches in terms of providing high spatial and temporal resolution mapping, sensor-based Variable Rate Application (VRA) mapping, the relation between spectral and thermal reflectance and different features of crop and soil. The other objective is to assess RS indices formed by choosing specific reflectance bands and identifying the correct spectral band to optimize classification techniques and analyze Proximal Optical Sensors (POSs) to control changes. The innovation of this paper can be defined as categorizing evaluation methodologies of precision irrigation (applying the right practice, at the right place, at the right time, with the right quantity) controlled by soil moisture levels and sensitiveness of crops to water stress, into pre-processing, processing (retrieval algorithms), and post-processing parts. Then, the main idea of this research is to analyze the error reasons and/or values in employing different approaches in three proposed parts reported by recent studies. Additionally, as an overview conclusion tried to decompose different approaches to optimizing indices, calibration methods for the sensors, thresholding and prediction models prone to errors, and improvements in classification accuracy for mapping changes.

Keywords: agricultural crops, crop water stress detection, irrigation scheduling, precision agriculture, remote sensing

Procedia PDF Downloads 42
119 Synthesis, Structure and Spectroscopic Properties of Oxo-centered Carboxylate-Bridged Triiron Complexes and a Deca Ferric Wheel

Authors: K. V. Ramanaiah, R. Jagan, N. N. Murthy

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Trinuclear oxo-centered carboxylate-bridged iron complexes, [Fe3(µ3-O)(µ2-O2CR)L¬3]+/0 (where R = alkyl or aryl; L = H2O, ROH, Py, solvent) have attracted tremendous attention because of their interesting structural and magnetic properties, exhibit mixed-valent trapped and de-trapped states, and have bioinorganic relevance. The presence of a trinuclear iron binding center has been implicated in the formation of both bacterial and human iron storage protein, Ft. They are used as precursors for the synthesis of models for the active-site structures of non-heme proteins, hemerythrin (Hr), methane monooxygenase (MMO) and polyiron storage protein, ferritin (Ft). Used as important building blocks for the design and synthesis of supramolecules this can exhibit single molecular magnetism (SMM). Such studies have often employed simple and compact carboxylate ligands and the use of bulky carboxylates is scarce. In the present study, we employed two different type of sterically hindered carboxylates and synthesized a series of novel oxo-centered, carboxylate-bridged triiron complexes of general formula [Fe3(O)(O2CCPh3)6L3]X (L = H2O, 1; py, 2; 4-NMe2py, 3; X = ClO4; L = CH3CN, 4; X = FeCl4) and [Fe3(O)(O2C-anth)6L3]X (L = H2O, 5; X = ClO4; L = CH3OH, 6; X = Cl). Along with complex [Fe(OMe)2(O2CCPh3)]10, 7 was prepared by the self-assemble of anhydrous FeCl3, sodium triphenylacetate and sodium methoxide at ratio of 1:1:2 in CH3OH. The Electronic absorption spectra of these complexes 1-6, in CH2Cl2 display weak bands at near FTIR region (970-1135 nm, ε > 15M-1cm-1). For complex 7, one broad band centered at ~670nm and also an additional intense charge transfer (L→M or O→M) bands between 300 to 550nm observed for all the complexes. Paramagnetic 1H NMR is introduced as a good probe for the characterization of trinuclear oxo - cantered iron compounds in solution when the L ligand coordinated to iron varies as: H2O, py, 4-NMe2py, and CH3OH. The solution state magnetic moment values calculated by using Evans method for all the complexes and also solid state magnetic moment value of complex, 7 was calculated by VSM method, which is comparable with solution state value. These all magnetic moment values indicate there is a spin exchange process through oxo and carboxylate bridges in between two irons (d5). The ESI-mass data complement the data obtained from single crystal X-ray structure. Further purity of the compounds was confirmed by elemental analysis. Finally, structural determination of complexes 1, 3, 4, 5, 6 and 7 were unambiguously conformed by single crystal x-ray studies.

Keywords: decanuclear, paramagnetic NMR, trinuclear, uv-visible

Procedia PDF Downloads 323
118 Molecular Detection of E. coli in Treated Wastewater and Well Water Samples Collected from Al Riyadh Governorate, Saudi Arabia

Authors: Hanouf A. S. Al Nuwaysir, Nadine Moubayed, Abir Ben Bacha, Islem Abid

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Consumption of waste water continues to cause significant problems for human health in both developed and developing countries. Many regulations have been implied by different world authorities controlling water quality for the presence of coliforms used as standard indicators of water quality deterioration and historically leading health protection concept. In this study, the European directive for the detection of Escherichia coli, ISO 9308-1, was applied to examine and monitor coliforms in water samples collected from Wadi Hanifa and neighboring wells, Riyadh governorate, kingdom of Saudi Arabia, which is used for irrigation and industrial purposes. Samples were taken from different locations for 8 months consecutively, chlorine concentration ranging from 0.1- 0.4 mg/l, was determined using the DPD FREE CHLORINE HACH kit. Water samples were then analyzed following the ISO protocol which relies on the membrane filtration technique (0.45µm, pore size membrane filter) and a chromogenic medium TTC, a lactose based medium used for the detection and enumeration of total coliforms and E.coli. Data showed that the number of bacterial isolates ranged from 60 to 300 colonies/100ml for well and surface water samples respectively; where higher numbers were attributed to the surface samples. Organisms which apparently ferment lactose on TTC agar plates, appearing as orange colonies, were selected and additionally cultured on EMB and MacConkey agar for a further differentiation among E.coli and coliform bacteria. Two additional biochemical tests (Cytochrome oxidase and indole from tryptophan) were also investigated to detect and differentiate the presence of E.coli from other coliforms, E. coli was identified in an average of 5 to 7colonies among 25 selected colonies.On the other hand, a more rapid, specific and sensitive analytical molecular detection namely single colony PCR was also performed targeting hha gene to sensitively detect E.coli, giving more accurate and time consuming identification of colonies considered presumptively as E.coli. Comparative methodologies, such as ultrafiltration and direct DNA extraction from membrane filters (MoBio, Grermany) were also applied; however, results were not as accurate as the membrane filtration, making it a technique of choice for the detection and enumeration of water coliforms, followed by sufficiently specific enzymatic confirmatory stage.

Keywords: coliform, cytochrome oxidase, hha primer, membrane filtration, single colony PCR

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117 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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116 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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115 Evaluation of Antimicrobial Properties of Lactic Acid Bacteria of Enterococcus Genus

Authors: Kristina Karapetyan, Flora Tkhruni, Tsovinar Balabekyan, Arevik Israyelyan, Tatyana Khachatryan

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The ability of the lactic acid bacteria (LAB) to prevent and cure a variety of diseases, their protective role against infections and colonization of pathogenic microorganisms in the digestive tract, has lead to the coining of the term probiotics or pro-life. LAB inhibiting the growth of pathogenic and food spoilage microorganisms, maintaining the nutritive quality and improving the shelf life of foods. They have also been used as flavor and texture producers. Enterococcus strains have been used for treatment of diseases such as diarrhea or antibiotic associated diarrhea, inflammatory pathologies that affect colon such as irritable bowel syndrome, or immune regulation, diarrhea caused by antibiotic treatments. The obtaining and investigation of biological properties of proteinoceous antibiotics, on the basis of probiotic LAB shown, that bacteriocins, metabiotics, and peptides of LAB represent bactericides have a broad range of activity and are excellent candidates for development of new prophylactic and therapeutic substances to complement or replace conventional antibiotics. The genotyping by 16S rRNA sequencing for LAB were used. Cell free culture broth (CFC) broth was purified by the Gel filtration method on the Sephadex Superfine G 25 resin. Antimicrobial activity was determined by spot-on-lawn method and expressed in arbitrary units (AU/ml). The diversity of multidrug-resistance (MDR) of pathogenic strains to antibiotics, most widely used for treatment of human diseases in the Republics of Armenia and Nagorno Karabakh were examined. It was shown, that difference of resistance of pathogens to antibiotics depends on their isolation sources. The influences of partially purified antimicrobial preparations (AMP), obtained from the different strains of Enterococcus genus on the growth of MDR pathogenic bacteria were investigated. It was shown, that bacteriocin containing partially purified preparations, obtained from different strains of Enterococcus faecium and durans species, possess bactericidal or bacteriostatic activity against antibiotic resistant intestinal, spoilage and food-borne pathogens such as Listeria monocytogenes, Staphylococcus aureus, E. coli, and Salmonella. Endemic strains of LAB, isolated from Matsoni made from donkey, buffalo and goat milk, shown broad spectrum of activity against food spoiling microorganisms, moulds and fungi, such as Salmonella sp., Esherichia coli, Aspergillus and Penicillium species. Highest activity against MDR pathogens shown bacteria, isolated from goat milk products. High stability of the investigated strains of the genus Enerococcus, isolated from samples of matsun from different regions of Nagorno-Karabakh (NKR) to the antibiotics was shown. The obtained data show high stability of the investigated different strains of the genus Enerococcus. The high genetic diversity in Enterococcus group suggests adaptations for specific mutations in different environments. Thus, endemic strains of LAB are able to produce bacteriocins with high and different inhibitory activity against broad spectrum of microorganisms isolated from different sources and belong to different taxonomic group. Prospect of the use of certain antimicrobial preparations against pathogenic strains is obvious. These AMP can be applied for long term use against different etiology antibiotic resistant pathogens for prevention or treatment of infectional diseases as an alternative to antibiotics.

Keywords: antimicrobial biopreparation, endemic lactic acid bacteria, intra-species diversity, multidrug resistance of pathogens

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114 Microbiological Analysis on Anatomical Specimens of Cats for Use in Veterinary Surgery

Authors: Raphael C. Zero, Marita V. Cardozo, Thiago A. S. S. Rocha, Mariana T. Kihara, Fernando A. Ávila, Fabrício S. Oliveira

Abstract:

There are several fixative and preservative solutions for use on cadavers, many of them using formaldehyde as the fixative or anatomical part preservative. In some countries, such as Brazil, this toxic agent has been increasingly restricted. The objective of this study was to microbiologically identify and quantify the key agents in tanks containing 96GL ethanol or sodium chloride solutions, used respectively as fixatives and preservatives of cat cadavers. Eight adult cat corpses, three females and five males, with an average weight of 4.3 kg, were used. After injection via the external common carotid artery (120 ml/kg, 95% 96GL ethyl alcohol and 5% pure glycerin), the cadavers were fixed in a plastic tank with 96GL ethanol for 60 days. After fixing, they were stored in a 30% sodium chloride aqueous solution for 120 days in a similar tank. Samples were collected at the start of the experiment - before the animals were placed in the ethanol tanks, and monthly thereafter. The bacterial count was performed by Pour Plate Method in BHI agar (Brain Heart Infusion) and the plates were incubated aerobically and anaerobically for 24h at 37ºC. MacConkey agar, SPS agar (Sulfite Polymyxin Sulfadizine) and MYP Agar Base were used to isolate the microorganisms. There was no microbial growth in the samples prior to alcohol fixation. After 30 days of fixation in the alcohol solution, total aerobic and anaerobic (<1.0 x 10 CFU/ml) were found and Pseudomonas sp., Staphylococcus sp., Clostridium sp. were the identified agents. After 60 days in the alcohol fixation solution, total aerobes (<1.0 x 10 CFU/ml) and total anaerobes (<2.2 x 10 CFU/mL) were found, and the identified agents were the same. After 30 days of storage in the aqueous solution of 30% sodium chloride, total aerobic (<5.2 x 10 CFU/ml) and total anaerobes (<3.7 x 10 CFU/mL) were found and the agents identified were Staphylococcus sp., Clostridium sp., and fungi. After 60 days of sodium chloride storage, total aerobic (<3.0 x 10 CFU / ml) and total anaerobes (<7.0 x 10 CFU/mL) were found and the identified agents remained the same: Staphylococcus sp., Clostridium sp., and fungi. The microbiological count was low and visual inspection did not reveal signs of contamination in the tanks. There was no strong odor or purification, which proved the technique to be microbiologically effective in fixing and preserving the cat cadavers for the four-month period in which they are provided to undergraduate students of University of Veterinary Medicine for surgery practice. All experimental procedures were approved by the Municipal Legal Department (protocol 02.2014.000027-1). The project was funded by FAPESP (protocol 2015-08259-9).

Keywords: anatomy, fixation, microbiology, small animal, surgery

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113 Comparative Characteristics of Bacteriocins from Endemic Lactic Acid Bacteria

Authors: K. Karapetyan, F. Tkhruni, A. Aghajanyan, T. S. Balabekyan, L. Arstamyan

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Introduction: Globalization of the food supply has created the conditions favorable for the emergence and spread of food-borne and especially dangerous pathogens (EDP) in developing countries. The fresh-cut fruit and vegetable industry is searching for alternatives to replace chemical treatments with biopreservative approaches that ensure the safety of the processed foods product. Antimicrobial compounds of lactic acid bacteria (LAB) possess bactericidal or bacteriostatic activity against intestinal pathogens, spoilage organisms and food-borne pathogens such as Listeria monocytogenes, Staphylococcus aureus and Salmonella. Endemic strains of LAB were isolated. The strains, showing broad spectrum of antimicrobial activity against food spoiling microorganisms, were selected. The genotyping by 16S rRNA sequencing, GS-PCR, RAPD PCR methods showed that they were presented by Lactobacillus rhamnosus109, L.plantarum 65, L.plantarum 66 and Enterococcus faecium 64 species. LAB are deposited in "Microbial Depository Center" (MDC) SPC "Armbiotechnology". Methods: LAB strains were isolated from different dairy products from rural households from the highland regions of Armenia. Serially diluted samples were spread on MRS (Merck, Germany) and hydrolyzed milk agar (1,2 % w/v). Single colonies from each LAB were individually inoculated in liquid MRS medium and incubated at 37oC for 24 hours. Culture broth with biomass was centrifuged at 10,000 g during 20 min for obtaining of cell free culture broth (CFC). The antimicrobial substances from CFC broth were purified by the combination of adsorption-desorption and ion-exchange chromatography methods. Separation of bacteriocins was performed using a HPLC method on "Avex ODS" C18 column. Mass analysis of peptides recorded on the device API 4000 in the electron ionization mode. The spot-on-lawn method on the test culture plated in the solid medium was applied. The antimicrobial activity is expressed in arbitrary units (AU/ml). Results. Purification of CFC broth of LAB allowed to obtain partially purified antimicrobial preparations which contains bacteriocins with broad spectrum of antimicrobial activity. Investigation of their main biochemical properties shown, that inhibitory activity of preparations is partially reduced after treatment with proteinase K, trypsin, pepsin, suggesting a proteinaceous nature of bacteriocin-like substances containing in CFC broth. Preparations preserved their activity after heat treatment (50-121 oC, 20 min) and were stable in the pH range 3–8. The results of SDS PAAG electrophoresis show that L.plantarum 66 and Ent.faecium 64 strains have one bacteriocin (BCN) with maximal antimicrobial activity with approximate molecular weight 2.0-3.0 kDa. From L.rhamnosus 109 two BCNs were obtained. Mass spectral analysis indicates that these bacteriocins have peptide bonds and molecular weight of BCN 1 and BCN 2 are approximately 1.5 kDa and 700 Da. Discussion: Thus, our experimental data shown, that isolated endemic strains of LAB are able to produce bacteriocins with high and different inhibitory activity against broad spectrum of microorganisms of different taxonomic group, such as Salmonella sp., Esherichia coli, Bacillus sp., L.monocytogenes, Proteus mirabilis, Staph. aureus, Ps. aeruginosa. Obtained results proved the perspectives for use of endemic strains in the preservation of foodstuffs. Acknowledgments: This work was realized with financial support of the Project Global Initiatives for Preliferation Prevention (GIPP) T2- 298, ISTC A-1866.

Keywords: antimicrobial activity, bacteriocins, endemic strains, food safety

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