Search results for: resistant microbial strains
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2550

Search results for: resistant microbial strains

120 Antimicrobial Efficacy of Some Antibiotics Combinations Tested against Some Molecular Characterized Multiresistant Staphylococcus Clinical Isolates, in Egypt

Authors: Nourhan Hussein Fanaki, Hoda Mohamed Gamal El-Din Omar, Nihal Kadry Moussa, Eva Adel Edward Farid

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The resistance of staphylococci to various antibiotics has become a major concern for health care professionals. The efficacy of the combinations of selected glycopeptides (vancomycin and teicoplanin) with gentamicin or rifampicin, as well as that of gentamicin/rifampicin combination, was studied against selected pathogenic staphylococcus isolated from Egypt. The molecular distribution of genes conferring resistance to these four antibiotics was detected among tested clinical isolates. Antibiotic combinations were studied using the checkerboard technique and the time-kill assay (in both the stationary and log phases). Induction of resistance to glycopeptides in staphylococci was tried in the absence and presence of diclofenac sodium as inducer. Transmission electron microscopy was used to study the effect of glycopeptides on the ultrastructure of the cell wall of staphylococci. Attempts were made to cure gentamicin resistance plasmids and to study the transfer of these plasmids by conjugation. Trials for the transformation of the successfully isolated gentamicin resistance plasmid to competent cells were carried out. The detection of genes conferring resistance to the tested antibiotics was performed using the polymerase chain reaction. The studied antibiotic combinations proved their efficacy, especially when tested during the log phase. Induction of resistance to glycopeptides in staphylococci was more promising in presence of diclofenac sodium, compared to its absence. Transmission electron microscopy revealed the thickening of bacterial cell wall in staphylococcus clinical isolates due to the presence of tested glycopeptides. Curing of gentamicin resistance plasmids was only successful in 2 out of 9 tested isolates, with a curing rate of 1 percent for each. Both isolates, when used as donors in conjugation experiments, yielded promising conjugation frequencies ranging between 5.4 X 10-2 and 7.48 X 10-2 colony forming unit/donor cells. Plasmid isolation was only successful in one out of the two tested isolates. However, low transformation efficiency (59.7 transformants/microgram plasmid DNA) of such plasmids was obtained. Negative regulators of autolysis, such as arlR, lytR and lrgB, as well as cell-wall associated genes, such as pbp4 and/or pbp2, were detected in staphylococcus isolates with reduced susceptibility to the tested glycopeptides. Concerning rifampicin resistance genes, rpoBstaph was detected in 75 percent of the tested staphylococcus isolates. It could be concluded that in vitro studies emphasized the usefulness of the combination of vancomycin or teicoplanin with gentamicin or rifampicin, as well as that of gentamicin with rifampicin, against staphylococci showing varying resistance patterns. However, further in vivo studies are required to ensure the safety and efficacy of such combinations. Diclofenac sodium can act as an inducer of resistance to glycopeptides in staphylococci. Cell-wall thickness is a major contributor to such resistance among them. Gentamicin resistance in these strains could be chromosomally or plasmid mediated. Multiple mutations in the rpoB gene could mediate staphylococcus resistance to rifampicin.

Keywords: glycopeptides, combinations, induction, diclofenac, transmission electron microscopy, polymerase chain reaction

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119 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

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118 Combined Pneumomediastinum and Pneumothorax Due to Hyperemesis Gravidarum

Authors: Fayez Hanna, Viet Tran

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A 20 years old lady- primigravida 6 weeks pregnant with unremarkable past history, presented to the emergency department at the Royal Hobart Hospital, Tasmania, Australia, with hyperemesis gravidarum associated with, dehydration and complicated with hematemesis and chest pain resistant. Accordingly, we conducted laboratory investigations which revealed: FBC: WBC 23.9, unremarkable U&E, LFT, lipase and her VBG showed a pH 7.4, pCo2 36.7, cK+ 3.2, cNa+ 142. The decision was made to do a chest X-ray (CXR) after explaining the risks/benefit of performing radiographic investigations during pregnancy and considering the patient's plan for the termination of the pregnancy as she was not ready for motherhood for shared decision-making and consent to look for pneumoperitoneum to suggest perforated viscus that might cause the hematemesis. However, the CXR showed pneumomediastinum but no evidence of pneumoperitoneum or pneumothorax. Consequently, a decision was made to proceed with CT oesophagography with imaging pre and post oral contrast administration to identify a potential oesophageal tear since it could not be excluded using a plain film of the CXR. The CT oesophagography could not find a leak for the administered oral contrast and thus, no oesophageal tear could be confirmed but could not exclude the Mallory-Weiss tear (lower oesophageal tear). Further, the CT oesophagography showed an extensive pneumomediastinum that could not be confirmed to be pulmonary in origin noting the presence of bilateral pulmonary interstitial emphysema and pneumothorax in the apex of the right lung that was small. The patient was admitted to the Emergency Department Inpatient Unit for monitoring, supportive therapy, and symptomatic management. Her hyperemesis was well controlled with ondansetron 8mg IV, metoclopramide 10mg IV, doxylamine 25mg PO, pyridoxine 25mg PO, esomeprazole 40mg IV and oxycodone 5mg PO was given for pain control and 2 litter of IV fluid. The patient was stabilized after 24 hours and discharged home on ondansetron 8mg every 8 hours whereas the patient had a plan for medical termination of pregnancy. Three weeks later, the patient represented with nausea and vomiting complicated by a frank hematemesis. Her observation chart showed HR 117- other vital signs were normal. Pathology showed WBC 14.3 with normal U&E and Hb. The patient was managed in the Emergency Department with the same previous regimen and was discharged home on same previous regimes. Five days later, she presented again with nausea, vomiting and hematemesis and was admitted under obstetrics and gynaecology for stabilization then discharged home with a plan for surgical termination of pregnancy after 3-days rather than the previously planned medical termination of pregnancy to avoid extension of potential oesophageal tear. The surgical termination and follow up period were uneventful. The case is considered rare as pneumomediastinum is a very rare complication of hyperemesis gravidarum where vomiting-induced barotrauma leads to a ruptured oesophagus and air leak into the mediastinum. However no rupture oesophagus in our case. Although the combination of pneumothorax and pneumomediastinum without oesophageal tear was reported only 8 times in the literature, but none of them was due to hyperemesis gravidarum.

Keywords: Pneumothorax, pneumomediastinum, hyperemesis gravidarum, pneumopericardium

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117 Treatment with Triton-X 100: An Enhancement Approach for Cardboard Bioprocessing

Authors: Ahlam Said Al Azkawi, Nallusamy Sivakumar, Saif Nasser Al Bahri

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Diverse approaches and pathways are under development with the determination to develop cellulosic biofuels and other bio-products eventually at commercial scale in “bio-refineries”; however, the key challenge is mainly the high level of complexity in processing the feedstock which is complicated and energy consuming. To overcome the complications in utilizing the naturally occurring lignocellulose biomass, using waste paper as a feedstock for bio-production may solve the problem. Besides being abundant and cheap, bioprocessing of waste paper has evolved in response to the public concern from rising landfill cost from shrinking landfill capacity. Cardboard (CB) is one of the major components of municipal solid waste and one of the most important items to recycle. Although 50-70% of cardboard constitute is known to be cellulose and hemicellulose, the presence of lignin around them cause hydrophobic cross-link which physically obstructs the hydrolysis by rendering it resistant to enzymatic cleavage. Therefore, pretreatment is required to disrupt this resistance and to enhance the exposure of the targeted carbohydrates to the hydrolytic enzymes. Several pretreatment approaches have been explored, and the best ones would be those can influence cellulose conversion rates and hydrolytic enzyme performance with minimal or less cost and downstream processes. One of the promising strategies in this field is the application of surfactants, especially non-ionic surfactants. In this study, triton-X 100 was used as surfactants to treat cardboard prior enzymatic hydrolysis and compare it with acid treatment using 0.1% H2SO4. The effect of the surfactant enhancement was evaluated through its effect on hydrolysis rate in respect to time in addition to evaluating the structural changes and modification by scanning electron microscope (SEM) and X-ray diffraction (XRD) and through compositional analysis. Further work was performed to produce ethanol from CB treated with triton-X 100 via separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF). The hydrolysis studies have demonstrated enhancement in saccharification by 35%. After 72 h of hydrolysis, a saccharification rate of 98% was achieved from CB enhanced with triton-X 100, while only 89 of saccharification achieved from acid pre-treated CB. At 120 h, the saccharification % exceeded 100 as reducing sugars continued to increase with time. This enhancement was not supported by any significant changes in the cardboard content as the cellulose, hemicellulose and lignin content remained same after treatment, but obvious structural changes were observed through SEM images. The cellulose fibers were clearly exposed with very less debris and deposits compared to cardboard without triton-X 100. The XRD pattern has also revealed the ability of the surfactant in removing calcium carbonate, a filler found in waste paper known to have negative effect on enzymatic hydrolysis. The cellulose crystallinity without surfactant was 73.18% and reduced to 66.68% rendering it more amorphous and susceptible to enzymatic attack. Triton-X 100 has proved to effectively enhance CB hydrolysis and eventually had positive effect on the ethanol yield via SSF. Treating cardboard with only triton-X 100 was a sufficient treatment to enhance the enzymatic hydrolysis and ethanol production.

Keywords: cardboard, enhancement, ethanol, hydrolysis, treatment, Triton-X 100

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116 In vitro Antimicrobial Resistance Pattern of Bovine Mastitis Bacteria in Ethiopia

Authors: Befekadu Urga Wakayo

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Introduction: Bacterial infections represent major human and animal health problems in Ethiopia. In the face of poor antibiotic regulatory mechanisms, development of antimicrobial resistance (AMR) to commonly used drugs has become a growing health and livelihood threat in the country. Monitoring and control of AMR demand close coloration between human and veterinary services as well as other relevant stakeholders. However, risk of AMR transfer from animal to human population’s remains poorly explored in Ethiopia. This systematic research literature review attempted to give an overview on AMR challenges of bovine mastitis bacteria in Ethiopia. Methodology: A web based research literature search and analysis strategy was used. Databases are considered including; PubMed, Google Scholar, Ethiopian Veterinary Association (EVA) and Ethiopian Society of Animal Production (ESAP). The key search terms and phrases were; Ethiopia, dairy, cattle, mastitis, bacteria isolation, antibiotic sensitivity and antimicrobial resistance. Ultimately, 15 research reports were used for the current analysis. Data extraction was performed using a structured Microsoft Excel format. Frequency AMR prevalence (%) was registered directly or calculated from reported values. Statistical analysis was performed on SPSS – 16. Variables were summarized by giving frequencies (n or %), Mean ± SE and demonstrative box plots. One way ANOVA and independent t test were used to evaluate variations in AMR prevalence estimates (Ln transformed). Statistical significance was determined at p < 0.050). Results: AMR in bovine mastitis bacteria was investigated in a total of 592 in vitro antibiotic sensitivity trials involving 12 different mastitis bacteria (including 1126 Gram positive and 77 Gram negative isolates) and 14 antibiotics. Bovine mastitis bacteria exhibited AMR to most of the antibiotics tested. Gentamycin had the lowest average AMR in both Gram positive (2%) and negative (1.8%) bacteria. Gram negative mastitis bacteria showed higher mean in vitro resistance levels to; Erythromycin (72.6%), Tetracycline (56.65%), Amoxicillin (49.6%), Ampicillin (47.6%), Clindamycin (47.2%) and Penicillin (40.6%). Among Gram positive mastitis bacteria higher mean in vitro resistance was observed in; Ampicillin (32.8%), Amoxicillin (32.6%), Penicillin (24.9%), Streptomycin (20.2%), Penicillinase Resistant Penicillin’s (15.4%) and Tetracycline (14.9%). More specifically, S. aurues exhibited high mean AMR against Penicillin (76.3%) and Ampicillin (70.3%) followed by Amoxicillin (45%), Streptomycin (40.6%), Tetracycline (24.5%) and Clindamycin (23.5%). E. coli showed high mean AMR to Erythromycin (78.7%), Tetracycline (51.5%), Ampicillin (49.25%), Amoxicillin (43.3%), Clindamycin (38.4%) and Penicillin (33.8%). Streptococcus spp. demonstrated higher (p =0.005) mean AMR against Kanamycin (> 20%) and full sensitivity (100%) to Clindamycin. Overall, mean Tetracycline (p = 0.013), Gentamycin (p = 0.001), Polymixin (p = 0.034), Erythromycin (p = 0.011) and Ampicillin (p = 0.009) resistance increased from the 2010’s than the 2000’s. Conclusion; the review indicated a rising AMR challenge among bovine mastitis bacteria in Ethiopia. Corresponding, public health implications demand a deeper, integrated investigation.

Keywords: antimicrobial resistance, dairy cattle, Ethiopia, Mastitis bacteria

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115 Application of NBR 14861: 2011 for the Design of Prestress Hollow Core Slabs Subjected to Shear

Authors: Alessandra Aparecida Vieira França, Adriana de Paula Lacerda Santos, Mauro Lacerda Santos Filho

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The purpose of this research i to study the behavior of precast prestressed hollow core slabs subjected to shear. In order to achieve this goal, shear tests were performed using hollow core slabs 26,5cm thick, with and without a concrete cover of 5 cm, without cores filled, with two cores filled and three cores filled with concrete. The tests were performed according to the procedures recommended by FIP (1992), the EN 1168:2005 and following the method presented in Costa (2009). The ultimate shear strength obtained within the tests was compared with the values of theoretical resistant shear calculated in accordance with the codes, which are being used in Brazil, noted: NBR 6118:2003 and NBR 14861:2011. When calculating the shear resistance through the equations presented in NBR 14861:2011, it was found that provision is much more accurate for the calculation of the shear strength of hollow core slabs than the NBR 6118 code. Due to the large difference between the calculated results, even for slabs without cores filled, the authors consulted the committee that drafted the NBR 14861:2011 and found that there is an error in the text of the standard, because the coefficient that is suggested, actually presents the double value than the needed one! The ABNT, later on, soon issued an amendment of NBR 14861:2011 with the necessary corrections. During the tests for the present study, it was confirmed that the concrete filling the cores contributes to increase the shear strength of hollow core slabs. But in case of slabs 26,5 cm thick, the quantity should be limited to a maximum of two cores filled, because most of the results for slabs with three cores filled were smaller. This confirmed the recommendation of NBR 14861:2011which is consistent with standard practice. After analyzing the configuration of cracking and failure mechanisms of hollow core slabs during the shear tests, strut and tie models were developed representing the forces acting on the slab at the moment of rupture. Through these models the authors were able to calculate the tensile stress acting on the concrete ties (ribs) and scaled the geometry of these ties. The conclusions of the research performed are the experiments results have shown that the mechanism of failure of the hollow-core slabs can be predicted using the strut-and-tie procedure, within a good range of accuracy. In addition, the needed of the correction of the Brazilian standard to review the correction factor σcp duplicated (in NBR14861/2011), and the limitation of the number of cores (Holes) to be filled with concrete, to increase the strength of the slab for the shear resistance. It is also suggested the increasing the amount of test results with 26.5 cm thick, and a larger range of thickness slabs, in order to obtain results of shear tests with cores concreted after the release of prestressing force. Another set of shear tests on slabs must be performed in slabs with cores filled and cover concrete reinforced with welded steel mesh for comparison with results of theoretical values calculated by the new revision of the standard NBR 14861:2011.

Keywords: prestressed hollow core slabs, shear, strut, tie models

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114 Rhizobium leguminosarum: Selecting Strain and Exploring Delivery Systems for White Clover

Authors: Laura Villamizar, David Wright, Claudia Baena, Marie Foxwell, Maureen O'Callaghan

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Leguminous crops can be self-sufficient for their nitrogen requirements when their roots are nodulated with an effective Rhizobium strain and for this reason seed or soil inoculation is practiced worldwide to ensure nodulation and nitrogen fixation in grain and forage legumes. The most widely used method of applying commercially available inoculants is using peat cultures which are coated onto seeds prior to sowing. In general, rhizobia survive well in peat, but some species die rapidly after inoculation onto seeds. The development of improved formulation methodology is essential to achieve extended persistence of rhizobia on seeds, and improved efficacy. Formulations could be solid or liquid. Most popular solid formulations or delivery systems are: wettable powders (WP), water dispersible granules (WG), and granules (DG). Liquid formulation generally are: suspension concentrates (SC) or emulsifiable concentrates (EC). In New Zealand, R. leguminosarum bv. trifolii strain TA1 has been used as a commercial inoculant for white clover over wide areas for many years. Seeds inoculation is carried out by mixing the seeds with inoculated peat, some adherents and lime, but rhizobial populations on stored seeds decline over several weeks due to a number of factors including desiccation and antibacterial compounds produced by the seeds. In order to develop a more stable and suitable delivery system to incorporate rhizobia in pastures, two strains of R. leguminosarum (TA1 and CC275e) and several formulations and processes were explored (peat granules, self-sticky peat for seed coating, emulsions and a powder containing spray dried microcapsules). Emulsions prepared with fresh broth of strain TA1 were very unstable under storage and after seed inoculation. Formulations where inoculated peat was used as the active ingredient were significantly more stable than those prepared with fresh broth. The strain CC275e was more tolerant to stress conditions generated during formulation and seed storage. Peat granules and peat inoculated seeds using strain CC275e maintained an acceptable loading of 108 CFU/g of granules or 105 CFU/g of seeds respectively, during six months of storage at room temperature. Strain CC275e inoculated on peat was also microencapsulated with a natural biopolymer by spray drying and after optimizing operational conditions, microparticles containing 107 CFU/g and a mean particle size between 10 and 30 micrometers were obtained. Survival of rhizobia during storage of the microcapsules is being assessed. The development of a stable product depends on selecting an active ingredient (microorganism), robust enough to tolerate some adverse conditions generated during formulation, storage, and commercialization and after its use in the field. However, the design and development of an adequate formulation, using compatible ingredients, optimization of the formulation process and selecting the appropriate delivery system, is possibly the best tool to overcome the poor survival of rhizobia and provide farmers with better quality inoculants to use.

Keywords: formulation, Rhizobium leguminosarum, storage stability, white clover

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113 Halloysite Based Adsorbents for Removing Pollutants from Water Reservoirs

Authors: Agata Chelminska, Joanna Goscianska

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The rapid growth of the world’s population and the resulting economic development have had an enormous influence on the environment. Multiple industrial processes generate huge amounts of wastewater containing dangerous substances, most of which are discharged into water bodies. These contaminants include pharmaceuticals and synthetic dyes. Regardless of the presence of wastewater treatment plants, a lot of pollutants cannot be easily eliminated by well-known technologies. Hence, more effective methods of removing resistant chemicals are being developed. Due to cost-effectiveness as well as the availability of a wide range of adsorbents, a large interest in the adsorption process as an alternative way of water purification has been observed. Clay minerals, e.g., halloysite, are one of the most researched natural adsorbents because of their availability, non-toxicity, high specific surface area, porosity, layered structure, and low cost. The negatively charged surface makes them ideal for binding cations and organic compounds. Halloysite can be subjected to modifications which enhance its adsorptive properties. The aim of the presented research was to apply pure and modified halloysite in removing particular pollutants (tetracycline, tartrazine, and phosphates) from aqueous solutions. Halloysite was modified with alcoholic and aqueous solutions of hexadecyltrimethylammonium bromide (CTAB) and urea in different concentrations and subsequently impregnated with lanthanum(III) chloride. Acidic and basic oxygen groups located on the surface of all materials were determined. Moreover, the adsorbents obtained were characterized by X-ray diffraction, low-temperature nitrogen adsorption, scanning, and transmission electron microscopy. The effectiveness of samples in tetracycline, tartrazine, and phosphates adsorption from the liquid phase was then studied in order to determine their potential application in eliminating contaminants from water reservoirs. Modifiers’ employment enabled obtaining materials that possess better adsorption properties, which makes them useful for removing various pollutants from water. Modifying the pure halloysite with CTAB and urea solutions and impregnating LaCl₃ led to the formation of acidic and basic oxygen functional groups on the surface. Their amount increases with an increasing percentage of lanthanum content. The acid-base properties of materials, as well as the type of functional groups that appear on their surface, have a significant influence on their sorption capacities towards antibiotics, dyes, and phosphate(V) anions. The selected contaminants adsorb onto the halloysite studied following the Langmuir type isotherm. The thermodynamic study indicated that the adsorption was a spontaneous and exothermic process. The adsorption equilibrium was rapidly attained after 120 min of contact time. Research showed that synthesized materials based on halloysite may be applied as adsorbents for antibiotics, organic dyes, and PO₄³- ions which are difficult to eliminate.

Keywords: adsorption processes, halloysite, minerals, water reservoirs pollutants

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112 Efficacy of DAPG Producing Fluorescent Pseudomonas for Enhancing Nutrient Use Efficacy, Bio-Control of Soil-Borne Diseases and Yield of Groundnut

Authors: Basavaraj Yenagi, P. Nagaraju, C. R. Patil

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Groundnut (Arachis hypohaea L.) is called as “King of oilseeds” and one of the most important food and cash crops in Indian subcontinent. Yield and quality of oil are negatively correlated with poor or imbalanced nutrition and constant exposure to both biotic and abiotic stress factors. Variety of diseases affect groundnut plant, most of them are caused by fungi and lead to severe yield loss. Imbalanced nutrition increases the concerns of environmental deterioration which includes soil fertility. Among different microbial antagonists, Pseudomonas is common member of the plant growth promoting rhizobacteria microflora present in the rhizosphere of groundnut. These are known to produce a beneficial effect on groundnut due to their high metabolic activity leading to the production of enzymes, exopolysaccharides, secondary metabolites, and antibiotics. The ability of pseudomonas lies on their ability to produce antibiotic metabolites such as 2, 4-diacetylphloroglucinol (DAPG). DAPG can inhibit the growth of fungal pathogens namely collar rot and stem rot and also increase the availability of plant nutrients through increased solubilization and uptake of nutrients. Hence, the present study was conducted for three consecutive years (2014 to 2016) in vertisol during the rainy season to assess the efficacy of DAPG producing fluorescent pseudomonas for enhancing nutrient use efficacy, bio-control of soil-borne diseases and yield of groundnut at University of Agricultural Sciences, Dharwad farm. The experiment was laid out in an RCBD with three replications and seven treatments. The mean of three years data revealed that the effect of DAPG-producing producing fluorescent pseudomonas enhanced groundnut yield, uptake of nitrogen and phosphorus and nutrient use efficiency and also found to be effective in bio-control of collar rot and stem rot incidence leading to increase pod yield of groundnut. Higher dry pod yield of groundnut was obtained with DAPG 2(3535 kg ha-1) closely followed by DAPG 4(3492 kg ha-1), FP 98(3443 kg ha-1), DAPG 1(3414 kg ha-1), FP 86(3361 kg ha-1) and Trichoderma spp. (3380 kg ha-1) over control(3173 kg ha-1). A similar trend was obtained with other growth and yield attributing parameters. N uptake ranged from 8.21 percent to FP 86 to 17.91 percent with DAPG 2 and P uptake ranged between 5.56 percent with FP 86 to 16.67 percent with DAPG 2 over control. The first year, there was no incidence of collar rot. During the second year, the control plot recorded 2.51 percent incidence and it ranged from 0.82 percent to 1.43 percent in different DAPG-producing fluorescent pseudomonas treatments. The similar trend was noticed in the third year with lower incidence. The stem rot incidence was recorded during all the three years. Mean data indicated that the control plot recorded 2.65 percent incidence and it ranged from 0.71 percent to 1.23 percent in different DAPG-producing fluorescent pseudomonas treatments. The increase in net monetary benefits ranged from Rs.5975 ha-1 to Rs.11407 ha 1 in different treatments. Hence, as a low-cost technology, seed treatment with available DAPG-producing fluorescent pseudomonas has a beneficial effect on groundnut for enhancing groundnut yield, nutrient use efficiency and bio-control of soil-borne diseases.

Keywords: groundnut, DAPG, fluorescent pseudomonas, nutrient use efficiency, collar rot, stem rot

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111 Selection and Preparation of High Performance, Natural and Cost-Effective Hydrogel as a Bio-Ink for 3D Bio-Printing and Organ on Chip Applications

Authors: Rawan Ashraf, Ahmed E. Gomaa, Gehan Safwat, Ayman Diab

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Background: Three-dimensional (3D) bio-printing has become a versatile and powerful method for generating a variety of biological constructs, including bone or extracellular matrix scaffolds endo- or epithelial, muscle tissue, as well as organoids. Aim of the study: Fabricate a low cost DIY 3D bio-printer to produce 3D bio-printed products such as anti-microbial packaging or multi-organs on chips. We demonstrate the alignment between two types of 3D printer technology (3D Bio-printer and DLP) on Multi-organ-on-a-chip (multi-OoC) devices fabrication. Methods: First, Design and Fabrication of the Syringe Unit for Modification of an Off-the-Shelf 3D Printer, then Preparation of Hydrogel based on natural polymers Sodium Alginate and Gelatin, followed by acquisition of the cell suspension, then modeling the desired 3D structure. Preparation for 3D printing, then Cell-free and cell-laden hydrogels went through the printing process at room temperature under sterile conditions and finally post printing curing process and studying the printed structure regards physical and chemical characteristics. The hard scaffold of the Organ on chip devices was designed and fabricated using the DLP-3D printer, following similar approaches as the Microfluidics system fabrication. Results: The fabricated Bio-Ink was based onHydrogel polymer mix of sodium alginate and gelatin 15% to 0.5%, respectively. Later the 3D printing process was conducted using a higher percentage of alginate-based hydrogels because of it viscosity and the controllable crosslinking, unlike the thermal crosslinking of Gelatin. The hydrogels were colored to simulate the representation of two types of cells. The adaption of the hard scaffold, whether for the Microfluidics system or the hard-tissues, has been acquired by the DLP 3D printers with fabricated natural bioactive essential oils that contain antimicrobial activity, followed by printing in Situ three complex layers of soft-hydrogel as a cell-free Bio-Ink to simulate the real-life tissue engineering process. The final product was a proof of concept for a rapid 3D cell culturing approaches that uses an engineered hard scaffold along with soft-tissues, thus, several applications were offered as products of the current prototype, including the Organ-On-Chip as a successful integration between DLP and 3D bioprinter. Conclusion: Multiple designs for the organ-on-a-chip (multi-OoC) devices have been acquired in our study with main focus on the low cost fabrication of such technology and the potential to revolutionize human health research and development. We describe circumstances in which multi-organ models are useful after briefly examining the requirement for full multi-organ models with a systemic component. Following that, we took a look at the current multi-OoC platforms, such as integrated body-on-a-chip devices and modular techniques that use linked organ-specific modules.

Keywords: 3d bio-printer, hydrogel, multi-organ on chip, bio-inks

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110 LaeA/1-Velvet Interplay in Aspergillus and Trichoderma: Regulation of Secondary Metabolites and Cellulases

Authors: Razieh Karimi Aghcheh, Christian Kubicek, Joseph Strauss, Gerhard Braus

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Filamentous fungi are of considerable economic and social significance for human health, nutrition and in white biotechnology. These organisms are dominant producers of a range of primary metabolites such as citric acid, microbial lipids (biodiesel) and higher unsaturated fatty acids (HUFAs). In particular, they produce also important but structurally complex secondary metabolites with enormous therapeutic applications in pharmaceutical industry, for example: cephalosporin, penicillin, taxol, zeranol and ergot alkaloids. Several fungal secondary metabolites, which are significantly relevant to human health do not only include antibiotics, but also e.g. lovastatin, a well-known antihypercholesterolemic agent produced by Aspergillus. terreus, or aflatoxin, a carcinogen produced by A. flavus. In addition to their roles for human health and agriculture, some fungi are industrially and commercially important: Species of the ascomycete genus Hypocrea spp. (teleomorph of Trichoderma) have been demonstrated as efficient producer of highly active cellulolytic enzymes. This trait makes them effective in disrupting and depolymerization of lignocellulosic materials and thus applicable tools in number of biotechnological areas as diverse as clothes-washing detergent, animal feed, and pulp and fuel productions. Fungal LaeA/LAE1 (Loss of aflR Expression A) homologs their gene products act at the interphase between secondary metabolisms, cellulase production and development. Lack of the corresponding genes results in significant physiological changes including loss of secondary metabolite and lignocellulose degrading enzymes production. At the molecular level, the encoded proteins are presumably methyltransferases or demethylases which act directly or indirectly at heterochromatin and interact with velvet domain proteins. Velvet proteins bind to DNA and affect expression of secondary metabolites (SMs) genes and cellulases. The dynamic interplay between LaeA/LAE1, velvet proteins and additional interaction partners is the key for an understanding of the coordination of metabolic and morphological functions of fungi and is required for a biotechnological control of the formation of desired bioactive products. Aspergilli and Trichoderma represent different biotechnologically significant species with significant differences in the LaeA/LAE1-Velvet protein machinery and their target proteins. We, therefore, performed a comparative study of the interaction partners of this machinery and the dynamics of the various protein-protein interactions using our robust proteomic and mass spectrometry techniques. This enhances our knowledge about the fungal coordination of secondary metabolism, cellulase production and development and thereby will certainly improve recombinant fungal strain construction for the production of industrial secondary metabolite or lignocellulose hydrolytic enzymes.

Keywords: cellulases, LaeA/1, proteomics, secondary metabolites

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109 Fermented Fruit and Vegetable Discard as a Source of Feeding Ingredients and Functional Additives

Authors: Jone Ibarruri, Mikel Manso, Marta Cebrián

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A high amount of food is lost or discarded in the World every year. In addition, in the last decades, an increasing demand of new alternative and sustainable sources of proteins and other valuable compounds is being observed in the food and feeding sectors and, therefore, the use of food by-products as nutrients for these purposes sounds very interesting from the environmental and economical point of view. However, the direct use of discarded fruit and vegetables that present, in general, a low protein content is not interesting as feeding ingredient except if they are used as a source of fiber for ruminants. Especially in the case of aquaculture, several alternatives to the use of fish meal and other vegetable protein sources have been extensively explored due to the scarcity of fish stocks and the unsustainability of fishing for these purposes. Fish mortality is also of great concern in this sector as this problem highly reduces their economic feasibility. So, the development of new functional and natural ingredients that could reduce the need for vaccination is also of great interest. In this work, several fermentation tests were developed at lab scale using a selected mixture of fruit and vegetable discards from a wholesale market located in the Basque Country to increase their protein content and also to produce some bioactive extracts that could be used as additives in aquaculture. Fruit and vegetable mixtures (60/40 ww) were centrifugated for humidity reduction and crushed to 2-5 mm particle size. Samples were inoculated with a selected Rhizopus oryzae strain and fermented for 7 days in controlled conditions (humidity between 65 and 75% and 28ºC) in Petri plates (120 mm) by triplicate. Obtained results indicated that the final fermented product presented a twofold protein content (from 13 to 28% d.w). Fermented product was further processed to determine their possible functionality as a feed additive. Extraction tests were carried out to obtain an ethanolic extract (60:40 ethanol: water, v.v) and remaining biomass that also could present applications in food or feed sectors. The extract presented a polyphenol content of about 27 mg GAE/gr d.w with antioxidant activity of 8.4 mg TEAC/g d.w. Remining biomass is mainly composed of fiber (51%), protein (24%) and fat (10%). Extracts also presented antibacterial activity according to the results obtained in Agar Diffusion and to the Minimum Inhibitory Concentration (MIC) tests determined against several food and fish pathogen strains. In vitro, digestibility was also assessed to obtain preliminary information about the expected effect of extraction procedure on fermented product digestibility. First results indicated that remaining biomass after extraction doesn´t seem to improve digestibility in comparison to the initial fermented product. These preliminary results show that fermented fruit and vegetables can be a useful source of functional ingredients for aquaculture applications and a substitute of other protein sources in the feeding sector. Further validation will be also carried out through “in vivo” tests with trout and bass.

Keywords: fungal solid state fermentation, protein increase, functional extracts, feed ingredients

Procedia PDF Downloads 44
108 An Adaptable Semi-Numerical Anisotropic Hyperelastic Model for the Simulation of High Pressure Forming

Authors: Daniel Tscharnuter, Eliza Truszkiewicz, Gerald Pinter

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High-quality surfaces of plastic parts can be achieved in a very cost-effective manner using in-mold processes, where e.g. scratch resistant or high gloss polymer films are pre-formed and subsequently receive their support structure by injection molding. The pre-forming may be done by high-pressure forming. In this process, a polymer sheet is heated and subsequently formed into the mold by pressurized air. Due to the heat transfer to the cooled mold the polymer temperature drops below its glass transition temperature. This ensures that the deformed microstructure is retained after depressurizing, giving the sheet its final formed shape. The development of a forming process relies heavily on the experience of engineers and trial-and-error procedures. Repeated mold design and testing cycles are however both time- and cost-intensive. It is, therefore, desirable to study the process using reliable computer simulations. Through simulations, the construction of the mold and the effect of various process parameters, e.g. temperature levels, non-uniform heating or timing and magnitude of pressure, on the deformation of the polymer sheet can be analyzed. Detailed knowledge of the deformation is particularly important in the forming of polymer films with integrated electro-optical functions. Care must be taken in the placement of devices, sensors and electrical and optical paths, which are far more sensitive to deformation than the polymers. Reliable numerical prediction of the deformation of the polymer sheets requires sophisticated material models. Polymer films are often either transversely isotropic or orthotropic due to molecular orientations induced during manufacturing. The anisotropic behavior affects the resulting strain field in the deformed film. For example, parts of the same shape but different strain fields may be created by varying the orientation of the film with respect to the mold. The numerical simulation of the high-pressure forming of such films thus requires material models that can capture the nonlinear anisotropic mechanical behavior. There are numerous commercial polymer grades for the engineers to choose from when developing a new part. The effort required for comprehensive material characterization may be prohibitive, especially when several materials are candidates for a specific application. We, therefore, propose a class of models for compressible hyperelasticity, which may be determined from basic experimental data and which can capture key features of the mechanical response. Invariant-based hyperelastic models with a reduced number of invariants are formulated in a semi-numerical way, such that the models are determined from a single uniaxial tensile tests for isotropic materials, or two tensile tests in the principal directions for transversely isotropic or orthotropic materials. The simulation of the high pressure forming of an orthotropic polymer film is finally done using an orthotropic formulation of the hyperelastic model.

Keywords: hyperelastic, anisotropic, polymer film, thermoforming

Procedia PDF Downloads 597
107 Achieving Sustainable Agriculture with Treated Municipal Wastewater

Authors: Reshu Yadav, Himanshu Joshi, S. K. Tripathi

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Fresh water is a scarce resource which is essential for humans and ecosystems, but its distribution is uneven. Agricultural production accounts for 70% of all surface water supplies. It is projected that against the expansion in the area equipped for irrigation by 0.6% per year, the global potential irrigation water demand would rise by 9.5% during 2021-25. This would, on one hand, have to compete against the sharply rising urban water demand. On the other, it would also have to face the fear of climate change, as temperatures rise and crop yields could drop from 10-30% in many large areas. The huge demand for irrigation combined with fresh water scarcity encourages to explore the reuse of wastewater as a resource. However, the use of such wastewater is often linked to the safety issues when used non judiciously or with poor safeguards while irrigating food crops. Paddy is one of the major crops globally and amongst the most important in South Asia and Africa. In many parts of the world, use of municipal wastewater has been promoted as a viable option in this regard. In developing and fast growing countries like India, regularly increasing wastewater generation rates may allow this option to be considered quite seriously. In view of this, a pilot field study was conducted at the Jagjeetpur Municipal Sewage treatment plant situated in the Haridwar town of Uttarakhand state, India. The objectives of the present study were to study the effect of treated wastewater on the production of various paddy varieties (Sharbati, PR-114, PB-1, Menaka, PB1121 and PB 1509) and emission of GHG gases (CO2, CH4 and N2O) as compared to the same varieties grown in the control plots irrigated with fresh water. Of late, the concept of water footprint assessment has emerged, which explains enumeration of various types of water footprints of an agricultural entity from its production to processing stages. Paddy, the most water demanding staple crop of Uttarakhand state, displayed a high green water footprint value of 2966.538 m3/ton. Most of the wastewater irrigated varieties displayed upto 6% increase in production, except Menaka and PB-1121, which showed a reduction in production (6% and 3% respectively), due to pest and insect infestation. The treated wastewater was observed to be rich in Nitrogen (55.94 mg/ml Nitrate), Phosphorus (54.24 mg/ml) and Potassium (9.78 mg/ml), thus rejuvenating the soil quality and not requiring any external nutritional supplements. Percentage increase of GHG gases on irrigation with treated municipal waste water as compared to control plots was observed as 0.4% - 8.6% (CH4), 1.1% - 9.2% (CO2), and 0.07% - 5.8% (N2O). The variety, Sharbati, displayed maximum production (5.5 ton/ha) and emerged as the most resistant variety against pests and insects. The emission values of CH4 ,CO2 and N2O were 729.31 mg/m2/d, 322.10 mg/m2/d and 400.21 mg/m2/d in water stagnant condition. This study highlighted a successful possibility of reuse of wastewater for non-potable purposes offering the potential for exploiting this resource that can replace or reduce existing use of fresh water sources in agricultural sector.

Keywords: greenhouse gases, nutrients, water footprint, wastewater irrigation

Procedia PDF Downloads 297
106 Sugarcane Trash Biochar: Effect of the Temperature in the Porosity

Authors: Gabriela T. Nakashima, Elias R. D. Padilla, Joao L. Barros, Gabriela B. Belini, Hiroyuki Yamamoto, Fabio M. Yamaji

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Biochar can be an alternative to use sugarcane trash. Biochar is a solid material obtained from pyrolysis, that is a biomass thermal degradation with low or no O₂ concentration. Pyrolysis transforms the carbon that is commonly found in other organic structures into a carbon with more stability that can resist microbial decomposition. Biochar has a versatility of uses such as soil fertility, carbon sequestration, energy generation, ecological restoration, and soil remediation. Biochar has a great ability to retain water and nutrients in the soil so that this material can improve the efficiency of irrigation and fertilization. The aim of this study was to characterize biochar produced from sugarcane trash in three different pyrolysis temperatures and determine the lowest temperature with the high yield and carbon content. Physical characterization of this biochar was performed to help the evaluation for the best production conditions. Sugarcane (Saccharum officinarum) trash was collected at Corredeira Farm, located in Ibaté, São Paulo State, Brazil. The farm has 800 hectares of planted area with an average yield of 87 t·ha⁻¹. The sugarcane varieties planted on the farm are: RB 855453, RB 867515, RB 855536, SP 803280, SP 813250. Sugarcane trash was dried and crushed into 50 mm pieces. Crucibles and lids were used to settle the sugarcane trash samples. The higher amount of sugarcane trash was added to the crucible to avoid the O₂ concentration. Biochar production was performed in three different pyrolysis temperatures (200°C, 325°C, 450°C) in 2 hours residence time in the muffle furnace. Gravimetric yield of biochar was obtained. Proximate analysis of biochar was done using ASTM E-872 and ABNT NBR 8112. Volatile matter and ash content were calculated by direct weight loss and fixed carbon content calculated by difference. Porosity measurement was evaluated using an automatic gas adsorption device, Autosorb-1, with CO₂ described by Nakatani. Approximately 0.5 g of biochar in 2 mm particle sizes were used for each measurement. Vacuum outgassing was performed as a pre-treatment in different conditions for each biochar temperature. The pore size distribution of micropores was determined using Horváth-Kawazoe method. Biochar presented different colors for each treatment. Biochar - 200°C presented a higher number of pieces with 10mm or more and did not present the dark black color like other treatments after 2 h residence time in muffle furnace. Also, this treatment had the higher content of volatiles and the lower amount of fixed carbon. In porosity analysis, while the temperature treatments increase, the amount of pores also increase. The increase in temperature resulted in a biochar with a better quality. The pores in biochar can help in the soil aeration, adsorption, water retention. Acknowledgment: This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – Brazil – PROAP-CAPES, PDSE and CAPES - Finance Code 001.

Keywords: proximate analysis, pyrolysis, soil amendment, sugarcane straw

Procedia PDF Downloads 176
105 Molecular Characterization, Host Plant Resistance and Epidemiology of Bean Common Mosaic Virus Infecting Cowpea (Vigna unguiculata L. Walp)

Authors: N. Manjunatha, K. T. Rangswamy, N. Nagaraju, H. A. Prameela, P. Rudraswamy, M. Krishnareddy

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The identification of virus in cowpea especially potyviruses is confusing. Even though there are several studies on viruses causing diseases in cowpea, difficult to distinguish based on symptoms and serological detection. The differentiation of potyviruses considering as a constraint, the present study is initiated for molecular characterization, host plant resistance and epidemiology of the BCMV infecting cowpea. The etiological agent causing cowpea mosaic was identified as Bean Common Mosaic Virus (BCMV) on the basis of RT-PCR and electron microscopy. An approximately 750bp PCR product corresponding to coat protein (CP) region of the virus and the presence of long flexuous filamentous particles measuring about 952 nm in size typical to genus potyvirus were observed under electron microscope. The characterized virus isolate genome had 10054 nucleotides, excluding the 3’ terminal poly (A) tail. Comparison of polyprotein of the virus with other potyviruses showed similar genome organization with 9 cleavage sites resulted in 10 functional proteins. The pairwise sequence comparison of individual genes, P1 showed most divergent, but CP gene was less divergent at nucleotide and amino acid level. A phylogenetic tree constructed based on multiple sequence alignments of the polyprotein nucleotide and amino acid sequences of cowpea BCMV and potyviruses showed virus is closely related to BCMV-HB. Whereas, Soybean variant of china (KJ807806) and NL1 isolate (AY112735) showed 93.8 % (5’UTR) and 94.9 % (3’UTR) homology respectively with other BCMV isolates. This virus transmitted to different leguminous plant species and produced systemic symptoms under greenhouse conditions. Out of 100 cowpea genotypes screened, three genotypes viz., IC 8966, V 5 and IC 202806 showed immune reaction in both field and greenhouse conditions. Single marker analysis (SMA) was revealed out of 4 SSR markers linked to BCMV resistance, M135 marker explains 28.2 % of phenotypic variation (R2) and Polymorphic information content (PIC) value of these markers was ranged from 0.23 to 0.37. The correlation and regression analysis showed rainfall, and minimum temperature had significant negative impact and strong relationship with aphid population, whereas weak correlation was observed with disease incidence. Path coefficient analysis revealed most of the weather parameters exerted their indirect contributions to the aphid population and disease incidence except minimum temperature. This study helps to identify specific gaps in knowledge for researchers who may wish to further analyse the science behind complex interactions between vector-virus and host in relation to the environment. The resistant genotypes identified are could be effectively used in resistance breeding programme.

Keywords: cowpea, epidemiology, genotypes, virus

Procedia PDF Downloads 206
104 Antibacterial Bioactive Glasses in Orthopedic Surgery and Traumatology

Authors: V. Schmidt, L. Janovák, N. Wiegand, B. Patczai, K. Turzó

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Large bone defects are not able to heal spontaneously. Bioactive glasses seem to be appropriate (bio)materials for bone reconstruction. Bioactive glasses are osteoconductive and osteoinductive, therefore, play a useful role in bony regeneration and repair. Because of their not optimal mechanical properties (e.g., brittleness, low bending strength, and fracture toughness), their applications are limited. Bioactive glass can be used as a coating material applied on metal surfaces. In this way -when using them as implants- the excellent mechanical properties of metals and the biocompatibility and bioactivity of glasses will be utilized. Furthermore, ion release effects of bioactive glasses regarding osteogenic and angiogenic responses have been shown. Silicate bioactive glasses (45S5 Bioglass) induce the release and exchange of soluble Si, Ca, P, and Na ions on the material surface. This will lead to special cellular responses inducing bone formation, which is favorable in the biointegration of the orthopedic prosthesis. The incorporation of other additional elements in the silicate network such as fluorine, magnesium, iron, silver, potassium, or zinc has been shown, as the local delivery of these ions is able to enhance specific cell functions. Although hip and knee prostheses present a high success rate, bacterial infections -mainly implant associated- are serious and frequent complications. Infection can also develop after implantation of hip prostheses, the elimination of which means more surgeries for the patient and additional costs for the clinic. Prosthesis-related infection is a severe complication of orthopedic surgery, which often causes prolonged illness, pain, and functional loss. While international efforts are made to reduce the risk of these infections, orthopedic surgical infections (SSIs) continue to occur in high numbers. It is currently estimated that up to 2.5% of primary hip and knee surgeries and up to 20% of revision arthroplasties are complicated by periprosthetic joint infection (PJIs). According to some authors, these numbers are underestimated, and they are also increasing. Staphylococcus aureus is the leading cause of both SSIs and PJIs, and the prevalence of methicillin-resistant S. aureus (MRSA) is on the rise, particularly in the United States. These deep infections lead to implant removal and consequently increase morbidity and mortality. The study targets this clinical problem using our experience so far with the Ag-doped polymer coatings on Titanium implants. Non-modified or modified (e.g., doped with antibacterial agents, like Ag) bioactive glasses could play a role in the prevention of infections or the therapy of infected tissues. Bioactive glasses have excellent biocompatibility, proved by in vitro cell culture studies of human osteoblast-like MG-63 cells. Ag-doped bioactive glass-scaffold has a good antibacterial ability against Escherichia coli and other bacteria. It may be concluded that these scaffolds have great potential in the prevention and therapy of implant-associated bone infection.

Keywords: antibacterial agents, bioactive glass, hip and knee prosthesis, medical implants

Procedia PDF Downloads 158
103 Ruminal Fermentation of Biologically Active Nitrate- and Nitro-Containing Forages

Authors: Robin Anderson, David Nisbet

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Nitrate, 3-nitro-1-propionic acid (NPA) and 3-nitro-1-propanol (NPOH) are biologically active chemicals that can accumulate naturally in rangeland grasses forages consumed by grazing cattle, sheep and goats. While toxic to livestock if accumulations and amounts consumed are high enough, particularly in animals having no recent exposure to the forages, these chemicals are known to be potent inhibitors of methane-producing bacteria inhabiting the rumen. Consequently, there is interest in examining their potential use as anti-methanogenic compounds to decrease methane emissions by grazing ruminants. Presently, rumen microbes, collected freshly from a cannulated Holstein cow maintained on 50:50 corn based concentrate:alfalfa diet were mixed (10 mL fluid) in 18 x 150 mm crimp top tubes with 0.5 of high nitrate-containing barley (Hordeum vulgare; containing 272 µmol nitrate per g forage dry matter), and NPA- or NPOH- containing milkvetch forages (Astragalus canadensis and Astragalus miser containing 80 and 174 soluble µmol NPA or NPOH/g forage dry matter respectively). Incubations containing 0.5 g alfalfa (Medicago sativa) were used as controls. Tubes (3 per each respective forage) were capped and incubated anaerobically (using oxygen free carbon dioxide) for 24 h at 39oC after which time amounts of total gas produced were measured via volume displacement and headspace samples were analyzed by gas chromatography to determine concentrations of hydrogen and methane. Fluid samples were analyzed by gas chromatography to measure accumulations of fermentation acids. A completely randomized analysis of variance revealed that the nitrate-containing barley and both the NPA- and the NPOH-containing milkvetches significantly decreased methane production, by > 50%, when compared to methane produced by populations incubated similarly with alfalfa (70.4 ± 3.6 µmol/ml incubation fluid). Accumulations of hydrogen, which are typically increased when methane production is inhibited, by incubations with the nitrate-containing barley and the NPA- and NPOH-containing milkvetches did not differ from accumulations observed in the alfalfa controls (0.09 ± 0.04 µmol/mL incubation fluid). Accumulations of fermentation acids produced in the incubations containing the high-nitrate barley and the NPA- and NPOH-containing milkvetches likewise did not differ from accumulations observed in incubations containing alfalfa (123.5 ± 10.8, 36.0 ± 3.0, 17.1 ± 1.5, 3.5 ± 0.3, 2.3 ± 0.2, 2.2 ± 0.2 µmol/mL incubation fluid for acetate, propionate, butyrate, valerate, isobutyrate, and isovalerate, respectively). This finding indicates the microbial populations did not compensate for the decreased methane production via compensatory changes in production of fermentative acids. Stoichiometric estimation of fermentation balance revealed that > 77% of reducing equivalents generated during fermentation of the forages were recovered in fermentation products and the recoveries did not differ between the alfalfa incubations and those with the high-nitrate barley or the NPA- or NPOH-containing milkvetches. Stoichiometric estimates of amounts of hexose fermented similarly did not differ between the nitrate-, NPA and NPOH-containing incubations and those with the alfalfa, averaging 99.6 ± 37.2 µmol hexose consumed/mL of incubation fluid. These results suggest that forages containing nitrate, NPA or NPOH may be useful to reduce methane emissions of grazing ruminants provided risks of toxicity can be effectively managed.

Keywords: nitrate, nitropropanol, nitropropionic acid, rumen methane emissions

Procedia PDF Downloads 102
102 Coping Strategies and Characterization of Vulnerability in the Perspective of Climate Change

Authors: Muhammad Umer Mehmood, Muhammad Luqman, Muhammad Yaseen, Imtiaz Hussain

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Climate change is an arduous fact, which could not be unheeded easily. It is a phenomenon which has brought a collection of challenges for the mankind. Scientists have found many of its negative impacts on the life of human being and the resources on which the life of humanity is dependent. There are many issues which are associated with the factor of prime importance in this study, 'climate change'. Whenever changes happen in nature, they strike the whole globe. Effects of these changes vary from region to region. Climate of every region of this globe is different from the other. Even within a state, country or the province has different climatic conditions. So it is mandatory that the response in that specific region and the coping strategy of this specific region should be according to the prevailing risk. In the present study, the objective was to assess the coping strategies and vulnerability of small landholders. So that a professional suggestion could be made to cope with the vulnerability factor of small farmers. The cross-sectional research design was used with the intervention of quantitative approach. The study was conducted in the Khanewal district, of Punjab, Pakistan. 120 small farmers were interviewed after randomized sampling from the population of respective area. All respondents were above the age of 15 years. A questionnaire was developed after keen observation of facts in the respective area. Content and face validity of the instrument was assessed with SPSS and experts in the field. Data were analyzed through SPSS using descriptive statistics. From the sample of 120, 81.67% of the respondents claimed that the environment is getting warmer and not fit for their present agricultural practices. 84.17% of the sample expressed serious concern that they are disturbed due to change in rainfall pattern and vulnerability towards the climatic effects. On the other hand, they expressed that they are not good at tackling the effects of climate change. Adaptation of coping strategies like change in cropping pattern, use of resistant varieties, varieties with minimum water requirement, intercropping and tree planting was low by more than half of the sample. From the sample 63.33% small farmers said that the coping strategies they adopt are not effective enough. The present study showed that subsistence farming, lack of marketing and overall infrastructure, lack of access to social security networks, limited access to agriculture extension services, inappropriate access to agrometeorological system, unawareness and access to scientific development and low crop yield are the prominent factors which are responsible for the vulnerability of small farmers. A comprehensive study should be conducted at national level so that a national policy could be formulated to cope with the dilemma in future with relevance to climate change. Mainstreaming and collaboration among the researchers and academicians could prove beneficiary in this regard the interest of national leaders’ does matter. Proper policies to avoid the vulnerability factors should be the top priority. The world is taking up this issue with full responsibility as should we, keeping in view the local situation.

Keywords: adaptation, coping strategies, climate change, Pakistan, small farmers, vulnerability

Procedia PDF Downloads 109
101 Activation of Apoptosis in the Midgut Epithelium of Spodoptera exigua Hübner (Lepidoptera: Noctuidae) Exposed to Various Cadmium Concentration

Authors: Magdalena Maria Rost-Roszkowska, Alina Chachulska-Żymełka, Monika Tarnawska, Maria Augustyniak, Alina Kafel, Agnieszka Babczyńska

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The digestive system of insects is composed of three distinct regions: fore-, mid- and hingut. The middle region (the midgut) is treated as one of the barriers which protects the organism against any stressors which originate from external environment, e.g. toxic metals. Such factors can activate the cell death in epithelial cells to preserve the entire tissue/organs against the degeneration. Different mechanisms involved in homeostasis maintenance have been described, but the studies of animals under field conditions do not give the opportunity to conclude about potential ability of subsequent generation to inherit the tolerance mechanisms. It is possible only by a multigenerational strain of an animal led under laboratory conditions, exposed to a selected toxic factor, present also in polluted ecosystems. The main purpose of the project was to check if changes, which appear in the midgut epithelium after Cd treatment, can be fixed during the following generations of insects with the special emphasis on apoptosis. As the animal for these studies we chose 5th larval stage of the beet armyworm Spodoptera exigua Hübner (Lepidoptera: Noctuidae), which is one of pest of many vegetable crops. Animals were divided into some experimental groups: K, Cd, KCd, Cd1, Cd2, Cd3. A control group (K) fed a standard diet, and was conducted for XX generations, a cadmium group (Cd), fed on standard diet supplemented with cadmium (44 mg Cd per kg of dry weight of food) for XXX generations. A reference Cd group (KCd) has been initiated: control insects were fed with Cd supplemented diet (44 mg Cd per kg of dry weight of food). Experimental groups Cd1, Cd2, Cd3 developed from the control one: 5 mg Cd per kg of dry weight of food, 10 mg Cd per kg of dry weight of food, 20 mg Cd per kg of dry weight of food. We were interested in the activation of apoptosis during following generations in all experimental groups. Therefore, during the 1st year of the experiment, the measurements were done for 6 generations in all experimental group. The intensity and the course of apoptosis have been examined using transmission electron microscope (TEM), confocal microscope and flow cytometry. During apoptosis the cell started to shrink, extracellular spaces appeared between digestive and neighboring cells, the nucleus achieved a lobular shape. Eventually, the apoptotic cells was discharged into the midgut lumen. A quantitative analysis revealed that the number of apoptotic cells depends significantly on the generation, tissue and cadmium concentration in the insect rearing medium. In the following 6 generations, we observed that the percentage of apoptotic cells in the midguts from cadmium-exposed groups decreased gradually according to the following order of strains: Cd1, Cd2, Cd3 and KCd. At the same time, it was still higher than the percentage of apoptotic cells in the same tissues of the insects from the control and multigenerational cadmium strain. The results of our studies suggest that changes caused by cadmium treatment were preserved during 6-generational development of lepidopteran larvae. The study has been financed by the National Science Centre Poland, grant no 2016/21/B/NZ8/00831.

Keywords: cadmium, cell death, digestive system, ultrastructure

Procedia PDF Downloads 193
100 Evaluation of the Incidence of Mycobacterium Tuberculosis Complex Associated with Soil, Hayfeed and Water in Three Agricultural Facilities in Amathole District Municipality in the Eastern Cape Province

Authors: Athini Ntloko

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Mycobacterium bovis and other species of Mycobacterium tuberculosis complex (MTBC) can result to a zoonotic infection known as Bovine tuberculosis (bTB). MTBC has members that may contaminate an extensive range of hosts, including wildlife. Diverse wild species are known to cause disease in domestic livestock and are acknowledged as TB reservoirs. It has been a main study worldwide to deliberate on bTB risk factors as a result and some studies focused on particular parts of risk factors such as wildlife and herd management. The significance of the study was to determine the incidence of Mycobacterium tuberculosis complex that is associated with soil, hayfeed and water. Questionnaires were administered to thirty (30) smallholding farm owners in the two villages (kwaMasele and Qungqwala) and three (3) three commercial farms (Fort Hare dairy farm, Middledrift dairy farm and Seven star dairy farm). Detection of M. tuberculosis complex was achieved by Polymerase Chain Reaction using primers for IS6110; whereas a genotypic drug resistance mutation was detected using Genotype MTBDRplus assays. Nine percent (9%) of respondents had more than 40 cows in their herd, while 60% reported between 10 and 20 cows in their herd. Relationship between farm size and vaccination for TB differed from forty one percent (41%) being the highest to the least five percent (5%). The highest number of respondents who knew about relationship between TB cases and cattle location was ninety one percent (91%). Approximately fifty one percent (51%) of respondents had knowledge about wild life access to the farms. Relationship between import of cattle and farm size ranged from nine percent (9%) to thirty five percent (35%). Cattle sickness in relation to farm size differed from forty three (43%) being the highest to the least three percent (3%); while thirty three percent (33%) of respondents had knowledge about health management. Respondents with knowledge about the occurrence of TB infections in farms were forty-eight percent (48%). The frequency of DNA isolation from samples ranged from the highest forty-five percent (45%) from water to the least twenty two percent (22%) from soil. Fort Hare dairy farm had the highest number of positive samples, forty four percent (44%) from water samples; whereas Middledrift dairy farm had the lowest positive from water, seventeen percent (17%). Twelve (22%) out of 55 isolates showed resistance to INH and RIF that is, multi-drug resistance (MDR) and nine percent (9%) were sensitive to either INH or RIF. The mutations at rpoB gene differed from 58% being the highest to the least (23%). Fifty seven percent (57%) of samples showed a S315T1 mutation while only 14% possessed a S531L in the katG gene. The highest inhA mutations were detected in T8A (80 %) and the least was observed in A16G (17%). The results of this study reveal that risk factors for bTB in cattle and dairy farm workers are a serious issue abound in the Eastern Cape of South Africa; with the possibility of widespread dissemination of multidrug resistant determinants in MTBC from the environment.

Keywords: hayfeed, isoniazid, multi-drug resistance, mycobacterium tuberculosis complex, polymerase chain reaction, rifampicin, soil, water

Procedia PDF Downloads 312
99 COVID-19: Potential Effects of Nutritional Factors on Inflammation Relief

Authors: Maryam Nazari

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COVID-19 is a respiratory disease triggered by the novel coronavirus, SARS-CoV-2, that has reached pandemic status today. Acute inflammation and immune cells infiltration into lung injuries result in multi-organ failure. The presence of other non-communicable diseases (NCDs) with systemic inflammation derived from COVID-19 may exacerbate the patient's situation and increase the risk for adverse effects and mortality. This pandemic is a novel situation and the scientific community at this time is looking for vaccines or drugs to treat the pathology. One of the biggest challenges is focused on reducing inflammation without compromising the correct immune response of the patient. In this regard, addressing the nutritional factors should not be overlooked not only as a matter of avoiding the presence of NCDs with severe infections but also as an adjunctive way to modulate the inflammatory status of the patients. Despite the pivotal role of nutrition in modifying immune response, due to the novelty of the COVID-19 disease, information about the effects of specific dietary agents is limited in this area. From the macronutrients point of view, protein deficiency (quantity or quality) has negative effects on the number of functional immunoglobulins and gut-associated lymphoid tissue (GALT). High biological value proteins or some amino acids like arginine and glutamine are well known for their ability to augment the immune system. Among lipids, fish oil has the ability to inactivate enveloped viruses, suppress pro-inflammatory prostaglandin production and block platelet-activating factors and their receptors. In addition, protectin D1, which is an Omega-3 PUFAs derivation, is a novel antiviral drug. So it seems that these fatty acids can reduce the severity and/or improve recovery of patients with COVID-19. Carbohydrates with lower glycemic index and fibers are associated with lower levels of inflammatory cytokines (CRP, TNF-α, and IL-6). Short-Chain Fatty acids not only exert a direct anti-inflammatory effect but also provide appropriate gut microbial, which is important in gastrointestinal issues related to COVID-19. From the micronutrients point of view, Vitamins A, C, D, E, iron, magnesium, zinc, selenium and copper play a vital role in the maintenance of immune function. Inadequate status in these nutrients may result in decreased resistance against COVID-19 infection. There are specific bioactive compounds in the diet that interact with the ACE2 receptor, which is the gateway for SARS and SARS-CoV-2, and thus controls the viral infection. Regarding this, the potential benefits of probiotics, resveratrol (a polyphenol found in grape), oleoylethanolamide (derived from oleic acid), and natural peroxisome proliferator-activated receptor γ agonists in foodstuffs (like curcumin, pomegranate, hot pepper) are suggested. Yet, it should be pointed out that most of these results have been reported in animal models and further human studies are needed to be verified.

Keywords: Covid-19, inflammation, nutrition, dietary agents

Procedia PDF Downloads 150
98 Impedimetric Phage-Based Sensor for the Rapid Detection of Staphylococcus aureus from Nasal Swab

Authors: Z. Yousefniayejahr, S. Bolognini, A. Bonini, C. Campobasso, N. Poma, F. Vivaldi, M. Di Luca, A. Tavanti, F. Di Francesco

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Pathogenic bacteria represent a threat to healthcare systems and the food industry because their rapid detection remains challenging. Electrochemical biosensors are gaining prominence as a novel technology for the detection of pathogens due to intrinsic features such as low cost, rapid response time, and portability, which make them a valuable alternative to traditional methodologies. These sensors use biorecognition elements that are crucial for the identification of specific bacteria. In this context, bacteriophages are promising tools for their inherent high selectivity towards bacterial hosts, which is of fundamental importance when detecting bacterial pathogens in complex biological samples. In this study, we present the development of a low-cost and portable sensor based on the Zeno phage for the rapid detection of Staphylococcus aureus. Screen-printed gold electrodes functionalized with the Zeno phage were used, and electrochemical impedance spectroscopy was applied to evaluate the change of the charge transfer resistance (Rct) as a result of the interaction with S. aureus MRSA ATCC 43300. The phage-based biosensor showed a linear range from 101 to 104 CFU/mL with a 20-minute response time and a limit of detection (LOD) of 1.2 CFU/mL under physiological conditions. The biosensor’s ability to recognize various strains of staphylococci was also successfully demonstrated in the presence of clinical isolates collected from different geographic areas. Assays using S. epidermidis were also carried out to verify the species-specificity of the phage sensor. We only observed a remarkable change of the Rct in the presence of the target S. aureus bacteria, while no substantial binding to S. epidermidis occurred. This confirmed that the Zeno phage sensor only targets S. aureus species within the genus Staphylococcus. In addition, the biosensor's specificity with respect to other bacterial species, including gram-positive bacteria like Enterococcus faecium and the gram-negative bacterium Pseudomonas aeruginosa, was evaluated, and a non-significant impedimetric signal was observed. Notably, the biosensor successfully identified S. aureus bacterial cells in a complex matrix such as a nasal swab, opening the possibility of its use in a real-case scenario. We diluted different concentrations of S. aureus from 108 to 100 CFU/mL with a ratio of 1:10 in the nasal swap matrices collected from healthy donors. Three different sensors were applied to measure various concentrations of bacteria. Our sensor indicated high selectivity to detect S. aureus in biological matrices compared to time-consuming traditional methods, such as enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), and radioimmunoassay (RIA), etc. With the aim to study the possibility to use this biosensor to address the challenge associated to pathogen detection, ongoing research is focused on the assessment of the biosensor’s analytical performances in different biological samples and the discovery of new phage bioreceptors.

Keywords: electrochemical impedance spectroscopy, bacteriophage, biosensor, Staphylococcus aureus

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97 Gene Expression Profiling of Iron-Related Genes of Pasteurella multocida Serotype A Strain PMTB2.1

Authors: Shagufta Jabeen, Faez Jesse Firdaus Abdullah, Zunita Zakaria, Nurulfiza Mat Isa, Yung Chie Tan, Wai Yan Yee, Abdul Rahman Omar

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Pasteurella multocida is associated with acute, as well as, chronic infections in avian and bovine such as pasteurellosis and hemorrhagic septicemia (HS) in cattle and buffaloes. Iron is one of the most important nutrients for pathogenic bacteria including Pasteurella and acts as a cofactor or prosthetic group in several essential enzymes and is needed for amino acid, pyrimidine, and DNA biosynthesis. In our recent study, we showed that 2% of Pasteurella multocida serotype A strain PMTB2.1 encode for iron regulating genes (Accession number CP007205.1). Genome sequencing of other Pasteurella multocida serotypes namely PM70 and HB01 also indicated up to 2.5% of the respective genome encode for iron regulating genes, suggesting that Pasteurella multocida genome comprises of multiple systems for iron uptake. Since P. multocida PMTB2.1 has more than 40 CDs out of 2097 CDs (approximately 2%), encode for iron-regulated. The gene expression profiling of four iron-regulating genes namely fbpb, yfea, fece and fur were characterized under iron-restricted environment. The P. multocida strain PMTB2.1 was grown in broth with and without iron chelating agent and samples were collected at different time points. Relative mRNA expression profile of these genes was determined using Taqman probe based real-time PCR assay. The data analysis, normalization with two house-keeping genes and the quantification of fold changes were carried out using Bio-Rad CFX manager software version 3.1. Results of this study reflect that iron reduced environment has significant effect on expression profile of iron regulating genes (p < 0.05) when compared to control (normal broth) and all evaluated genes act differently with response to iron reduction in media. The highest relative fold change of fece gene was observed at early stage of treatment indicating that PMTB2.1 may utilize its periplasmic protein at early stage to acquire iron. Furthermore, down-regulation expression of fece with the elevated expression of other genes at later time points suggests that PMTB2.1 control their iron requirements in response to iron availability by down-regulating the expression of iron proteins. Moreover, significantly high relative fold change (p ≤ 0.05) of fbpb gene is probably associated with the ability of P. multocida to directly use host iron complex such as hem, hemoglobin. In addition, the significant increase (p ≤ 0.05) in fbpb and yfea expressions also reflects the utilization of multiple iron systems in P. multocida strain PMTB2.1. The findings of this study are very much important as relative scarcity of free iron within hosts creates a major barrier to microbial growth inside host and utilization of outer-membrane proteins system in iron acquisition probably occurred at early stage of infection with P. multocida. In conclusion, the presence and utilization of multiple iron system in P. multocida strain PMTB2.1 revealed the importance of iron in the survival of P. multocida.

Keywords: iron-related genes, real-time PCR, gene expression profiling, fold changes

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96 Approaches to Inducing Obsessional Stress in Obsessive-Compulsive Disorder (OCD): An Empirical Study with Patients Undergoing Transcranial Magnetic Stimulation (TMS) Therapy

Authors: Lucia Liu, Matthew Koziol

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Obsessive-compulsive disorder (OCD), a long-lasting anxiety disorder involving recurrent, intrusive thoughts, affects over 2 million adults in the United States. Transcranial magnetic stimulation (TMS) stands out as a noninvasive, cutting-edge therapy that has been shown to reduce symptoms in patients with treatment-resistant OCD. The Food and Drug Administration (FDA) approved protocol pairs TMS sessions with individualized symptom provocation, aiming to improve the susceptibility of brain circuits to stimulation. However, limited standardization or guidance exists on how to conduct symptom provocation and which methods are most effective. This study aims to compare the effect of internal versus external techniques to induce obsessional stress in a clinical setting during TMS therapy. Two symptom provocation methods, (i) Asking patients thought-provoking questions about their obsessions (internal) and (ii) Requesting patients to perform obsession-related tasks (external), were employed in a crossover design with repeated measurement. Thirty-six treatments of NeuroStar TMS were administered to each of two patients over 8 weeks in an outpatient clinic. Patient One received 18 sessions of internal provocation followed by 18 sessions of external provocation, while Patient Two received 18 sessions of external provocation followed by 18 sessions of internal provocation. The primary outcome was the level of self-reported obsessional stress on a visual analog scale from 1 to 10. The secondary outcome was self-reported OCD severity, collected biweekly in a four-level Likert-scale (1 to 4) of bad, fair, good and excellent. Outcomes were compared and tested between provocation arms through repeated measures ANOVA, accounting for intra-patient correlations. Ages were 42 for Patient One (male, White) and 57 for Patient Two (male, White). Both patients had similar moderate symptoms at baseline, as determined through the Yale-Brown Obsessive Compulsive Scale (YBOCS). When comparing obsessional stress induced across the two arms of internal and external provocation methods, the mean (SD) was 6.03 (1.18) for internal and 4.01 (1.28) for external strategies (P=0.0019); ranges were 3 to 8 for internal and 2 to 8 for external strategies. Internal provocation yielded 5 (31.25%) bad, 6 (33.33%) fair, 3 (18.75%) good, and 2 (12.5%) excellent responses for OCD status, while external provocation yielded 5 (31.25%) bad, 9 (56.25%) fair, 1 (6.25%) good, and 1 (6.25%) excellent responses (P=0.58). Internal symptom provocation tactics had a significantly stronger impact on inducing obsessional stress and led to better OCD status (non-significant). This could be attributed to the fact that answering questions may prompt patients to reflect more on their lived experiences and struggles with OCD. In the future, clinical trials with larger sample sizes are warranted to validate this finding. Results support the increased integration of internal methods into structured provocation protocols, potentially reducing the time required for provocation and achieving greater treatment response to TMS.

Keywords: obsessive-compulsive disorder, transcranial magnetic stimulation, mental health, symptom provocation

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95 Biosynthesis of Silver Nanoparticles Using Zataria multiflora Extract, and Study of Antibacterial Effects on UTI Bacteria (MDR)

Authors: Mohammad Hossein Pazandeh, Monir Doudi, Sona Rostampour Yasouri

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Irregular consumption of current antibiotic makes increases of antibiotic resistance between urin pathogens on all worlds. This study selected based on this great community problem. The aim of this study was the biosynthesis of silver nanoparticles from Zataria multiflora extract and then to investigate its antibacterial effect on gram-negative bacilli common in Urinary Tract Infections (UTI) and MDR. The plant used in the present research was Zataria multiflora whose extract was prepared through Soxhlet extraction method. Green synthesis condition of silver nanoparticles was investigated in terms of three parameters including the extract amount, concentration of silver nitrate salt, and temperature. The seizes of nanoparticles were determined by Zetasizer. In order to identify synthesized silver nanoparticles Transmission Electron Microscopy (TEM) and X-ray Diffraction (XRD) methods were used. For evaluating the antibacterial effects of nanoparticles synthesized through biological method different concentrations of silver nanoparticles were studied on 140 cases of Muliple Drug Resistance (MDR) bacteria strains Escherichia coli, Klebsiella pneumoniae, Enterobacter aerogenes, Proteus vulgaris,Citrobacter freundii, Acinetobacter bumanii and Pseudomonas aeruginosa, (each genus of bacteria, 20 samples), which all were MDR and cause urinary tract infections , for identification of bacteria were used of Polymerase Chain Reaction (PCR) test and laboratory methods (Agar well diffusion and Microdilution methods) to assess their sensitivity to Nanoparticles. The data were analyzed using SPSS software by nonparametric Kruskal-Wallis and Mann-Whitney tests. Significant results were found about the effects of silver nitrate concentration, different amounts of Zataria multiflora extract, and temperature on nanoparticles; that is, by increasing the concentration of silver nitrate, extract amount, and temperature, the sizes of synthesized nanoparticles declined. However, the effect of above mentioned factors on particles diffusion index was not significant. Based on the TEM results, particles were mainly spherical shape with a diameter range of 25 to 50 nm. The results of XRD Analysis indicated the formation of Nanostructures and Nanocrystals of silver.. The obtained results of antibacterial effects of different concentrations of silver nanoparticles on according to agar well diffusion and microdilution method, biologically synthesized nanoparticles showed 1000 mg /ml highest and lowest mean inhibition zone diameter in E.coli , Acinetobacter bumanii 23 and 15mm, respectively. MIC was observed for all of bacteria 125mg/ml and for Acinetobacter bumanii 250mg/ml.Comparing the growth inhibitory effect of chemically synthesized Nanoparticles and biologically synthesized Nanoparticles showed that in the chemical method the highest growth inhibition belonged to the concentration of 62.5 mg /ml. The inhibitory effect on the growth all of bacteria causes of urine infection and MDR was observed and by increasing silver ion concentration in Nanoparticles, antibacterial activity increased. Generally, the biological synthesis can be considered an efficient way not only in making Nanoparticles but also for having anti-bacterial properties. It is more biocompatible and may be possess less toxicity than the Nanoparticles synthesized chemically.

Keywords: biosynthesis, MDR bacteria, silver nanoparticles, UTI

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94 The Use of Vasopressin in the Management of Severe Traumatic Brain Injury: A Narrative Review

Authors: Nicole Selvi Hill, Archchana Radhakrishnan

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Introduction: Traumatic brain injury (TBI) is a leading cause of mortality among trauma patients. In the management of TBI, the main principle is avoiding cerebral ischemia, as this is a strong determiner of neurological outcomes. The use of vasoactive drugs, such as vasopressin, has an important role in maintaining cerebral perfusion pressure to prevent secondary brain injury. Current guidelines do not suggest a preferred vasoactive drug to administer in the management of TBI, and there is a paucity of information on the therapeutic potential of vasopressin following TBI. Vasopressin is also an endogenous anti-diuretic hormone (AVP), and pathways mediated by AVP play a large role in the underlying pathological processes of TBI. This creates an overlap of discussion regarding the therapeutic potential of vasopressin following TBI. Currently, its popularity lies in vasodilatory and cardiogenic shock in the intensive care setting, with increasing support for its use in haemorrhagic and septic shock. Methodology: This is a review article based on a literature review. An electronic search was conducted via PubMed, Cochrane, EMBASE, and Google Scholar. The aim was to identify clinical studies looking at the therapeutic administration of vasopressin in severe traumatic brain injury. The primary aim was to look at the neurological outcome of patients. The secondary aim was to look at surrogate markers of cerebral perfusion measurements, such as cerebral perfusion pressure, cerebral oxygenation, and cerebral blood flow. Results: Eight papers were included in the final number. Three were animal studies; five were human studies, comprised of three case reports, one retrospective review of data, and one randomised control trial. All animal studies demonstrated the benefits of vasopressors in TBI management. One animal study showed the superiority of vasopressin in reducing intracranial pressure and increasing cerebral oxygenation over a catecholaminergic vasopressor, phenylephrine. All three human case reports were supportive of vasopressin as a rescue therapy in catecholaminergic-resistant hypotension. The retrospective review found vasopressin did not increase cerebral oedema in TBI patients compared to catecholaminergic vasopressors; and demonstrated a significant reduction in the requirements of hyperosmolar therapy in patients that received vasopressin. The randomised control trial results showed no significant differences in primary and secondary outcomes between TBI patients receiving vasopressin versus those receiving catecholaminergic vasopressors. Apart from the randomised control trial, the studies included are of low-level evidence. Conclusion: Studies favour vasopressin within certain parameters of cerebral function compared to control groups. However, the neurological outcomes of patient groups are not known, and animal study results are difficult to extrapolate to humans. It cannot be said with certainty whether vasopressin’s benefits stand above usage of other vasoactive drugs due to the weaknesses of the evidence. Further randomised control trials, which are larger, standardised, and rigorous, are required to improve knowledge in this field.

Keywords: catecholamines, cerebral perfusion pressure, traumatic brain injury, vasopressin, vasopressors

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93 Leveraging Advanced Technologies and Data to Eliminate Abandoned, Lost, or Otherwise Discarded Fishing Gear and Derelict Fishing Gear

Authors: Grant Bifolchi

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As global environmental problems continue to have highly adverse effects, finding long-term, sustainable solutions to combat ecological distress are of growing paramount concern. Ghost Gear—also known as abandoned, lost or otherwise discarded fishing gear (ALDFG) and derelict fishing gear (DFG)—represents one of the greatest threats to the world’s oceans, posing a significant hazard to human health, livelihoods, and global food security. In fact, according to the UN Food and Agriculture Organization (FAO), abandoned, lost and discarded fishing gear represents approximately 10% of marine debris by volume. Around the world, many governments, governmental and non-profit organizations are doing their best to manage the reporting and retrieval of nets, lines, ropes, traps, floats and more from their respective bodies of water. However, these organizations’ ability to effectively manage files and documents about the environmental problem further complicates matters. In Ghost Gear monitoring and management, organizations face additional complexities. Whether it’s data ingest, industry regulations and standards, garnering actionable insights into the location, security, and management of data, or the application of enforcement due to disparate data—all of these factors are placing massive strains on organizations struggling to save the planet from the dangers of Ghost Gear. In this 90-minute educational session, globally recognized Ghost Gear technology expert Grant Bifolchi CET, BBA, Bcom, will provide real-world insight into how governments currently manage Ghost Gear and the technology that can accelerate success in combatting ALDFG and DFG. In this session, attendees will learn how to: • Identify specific technologies to solve the ingest and management of Ghost Gear data categories, including type, geo-location, size, ownership, regional assignment, collection and disposal. • Provide enhanced access to authorities, fisheries, independent fishing vessels, individuals, etc., while securely controlling confidential and privileged data to globally recognized standards. • Create and maintain processing accuracy to effectively track ALDFG/DFG reporting progress—including acknowledging receipt of the report and sharing it with all pertinent stakeholders to ensure approvals are secured. • Enable and utilize Business Intelligence (BI) and Analytics to store and analyze data to optimize organizational performance, maintain anytime-visibility of report status, user accountability, scheduling, management, and foster governmental transparency. • Maintain Compliance Reporting through highly defined, detailed and automated reports—enabling all stakeholders to share critical insights with internal colleagues, regulatory agencies, and national and international partners.

Keywords: ghost gear, ALDFG, DFG, abandoned, lost or otherwise discarded fishing gear, data, technology

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92 Edible Active Antimicrobial Coatings onto Plastic-Based Laminates and Its Performance Assessment on the Shelf Life of Vacuum Packaged Beef Steaks

Authors: Andrey A. Tyuftin, David Clarke, Malco C. Cruz-Romero, Declan Bolton, Seamus Fanning, Shashi K. Pankaj, Carmen Bueno-Ferrer, Patrick J. Cullen, Joe P. Kerry

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Prolonging of shelf-life is essential in order to address issues such as; supplier demands across continents, economical profit, customer satisfaction, and reduction of food wastage. Smart packaging solutions presented in the form of naturally occurred antimicrobially-active packaging may be a solution to these and other issues. Gelatin film forming solution with adding of natural sourced antimicrobials is a promising tool for the active smart packaging. The objective of this study was to coat conventional plastic hydrophobic packaging material with hydrophilic antimicrobial active beef gelatin coating and conduct shelf life trials on beef sub-primal cuts. Minimal inhibition concentration (MIC) of Caprylic acid sodium salt (SO) and commercially available Auranta FV (AFV) (bitter oranges extract with mixture of nutritive organic acids) were found of 1 and 1.5 % respectively against bacterial strains Bacillus cereus, Pseudomonas fluorescens, Escherichia coli, Staphylococcus aureus and aerobic and anaerobic beef microflora. Therefore SO or AFV were incorporated in beef gelatin film forming solution in concentration of two times of MIC which was coated on a conventional plastic LDPE/PA film on the inner cold plasma treated polyethylene surface. Beef samples were vacuum packed in this material and stored under chilling conditions, sampled at weekly intervals during 42 days shelf life study. No significant differences (p < 0.05) in the cook loss was observed among the different treatments compared to control samples until the day 29. Only for AFV coated beef sample it was 3% higher (37.3%) than the control (34.4 %) on the day 36. It was found antimicrobial films did not protect beef against discoloration. SO containing packages significantly (p < 0.05) reduced Total viable bacterial counts (TVC) compared to the control and AFV samples until the day 35. No significant reduction in TVC was observed between SO and AFV films on the day 42 but a significant difference was observed compared to control samples with a 1.40 log of bacteria reduction on the day 42. AFV films significantly (p < 0.05) reduced TVC compared to control samples from the day 14 until the day 42. Control samples reached the set value of 7 log CFU/g on day 27 of testing, AFV films did not reach this set limit until day 35 and SO films until day 42 of testing. The antimicrobial AFV and SO coated films significantly prolonged the shelf-life of beef steaks by 33 or 55% (on 7 and 14 days respectively) compared to control film samples. It is concluded antimicrobial coated films were successfully developed by coating the inner polyethylene layer of conventional LDPE/PA laminated films after plasma surface treatment. The results indicated that the use of antimicrobial active packaging coated with SO or AFV increased significantly (p < 0.05) the shelf life of the beef sub-primal. Overall, AFV or SO containing gelatin coatings have the potential of being used as effective antimicrobials for active packaging applications for muscle-based food products.

Keywords: active packaging, antimicrobials, edible coatings, food packaging, gelatin films, meat science

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91 Aerobic Biodegradation of a Chlorinated Hydrocarbon by Bacillus Cereus 2479

Authors: Srijata Mitra, Mobina Parveen, Pranab Roy, Narayan Chandra Chattopadhyay

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Chlorinated hydrocarbon can be a major pollution problem in groundwater as well as soil. Many people interact with these chemicals on daily accidentally or by professionally in the laboratory. One of the most common sources for Chlorinated hydrocarbon contamination of soil and groundwater are industrial effluents. The wide use and discharge of Trichloroethylene (TCE), a volatile chlorohydrocarbon from chemical industry, led to major water pollution in rural areas. TCE is an mainly used as an industrial metal degreaser in industries. Biotransformation of TCE to the potent carcinogen vinyl chloride (VC) by consortia of anaerobic bacteria might have role for the above purpose. For these reasons, the aim of current study was to isolate and characterized the genes involved in TCE metabolism and also to investigate the in silico study of those genes. To our knowledge, only one aromatic dioxygenase system, the toluene dioxygenase in Pseudomonas putida F1 has been shown to be involved in TCE degradation. This is first instance where Bacillus cereus group being used in biodegradation of trichloroethylene. A novel bacterial strain 2479 was isolated from oil depot site at Rajbandh, Durgapur (West Bengal, India) by enrichment culture technique. It was identified based on polyphasic approach and ribotyping. The bacterium was gram positive, rod shaped, endospore forming and capable of degrading trichloroethylene as the sole carbon source. On the basis of phylogenetic data and Fatty Acid Methyl Ester Analysis, strain 2479 should be placed within the genus Bacillus and species cereus. However, the present isolate (strain 2479) is unique and sharply different from the usual Bacillus strains in its biodegrading nature. Fujiwara test was done to estimate that the strain 2479 could degrade TCE efficiently. The gene for TCE biodegradation was PCR amplified from genomic DNA of Bacillus cereus 2479 by using todC1 gene specific primers. The 600bp amplicon was cloned into expression vector pUC I8 in the E. coli host XL1-Blue and expressed under the control of lac promoter and nucleotide sequence was determined. The gene sequence was deposited at NCBI under the Accession no. GU183105. In Silico approach involved predicting the physico-chemical properties of deduced Tce1 protein by using ProtParam tool. The tce1 gene contained 342 bp long ORF encoding 114 amino acids with a predicted molecular weight 12.6 kDa and the theoretical pI value of the polypeptide was 5.17, molecular formula: C559H886N152O165S8, total number of atoms: 1770, aliphatic index: 101.93, instability index: 28.60, Grand Average of Hydropathicity (GRAVY): 0.152. Three differentially expressed proteins (97.1, 40 and 30 kDa) were directly involved in TCE biodegradation, found to react immunologically to the antibodies raised against TCE inducible proteins in Western blot analysis. The present study suggested that cloned gene product (TCE1) was capable of degrading TCE as verified chemically.

Keywords: cloning, Bacillus cereus, in silico analysis, TCE

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