Search results for: Bacillus cereus strain JMG-01

Authors: Suruchi Shukla, Shantanu Prakash, Amita Jain

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Introduction: Arboviral diseases are one of the most common causes of viral hemorrhagic fever (VHF). Of which, Dengue and Chikungunya pose a significant health problem in India. Arbovirus has a tendency to cross the territories and emerge in the new region. Considering the above issues, in the current study active surveillance was conducted among viral hemorrhagic fever (VHF) cases reported from Uttar Pradesh (UP), India. We studied the arboviral etiology of VHF; mainly Dengue, Chikungunya, and ZIKA. Methods: Clinical samples of 465 suspected VHF cases referred to tertiary care referral center of UP, India were enrolled in the study during a period from 15th May 2016 to 9th March 2018. Serum specimens were collected and analyzed for the presence of Dengue, Chikungunya, and ZIKA either by serology and/or by molecular assays. Results: Of all tested, 165 (35.4%) cases were positive for either Dengue or Chikungunya. Dengue (21.2%) was found to be the most prevalent, followed by Chikungunya, (6.6%). None of the cases tested positive for ZIKA virus. Serum samples of 35 (7.5%) cases were positive for both Dengue and Chikungunya. DEN-2 serotype was the most predominant serotype. Phylogenetic and sequence analysis of DEN-2 strains showed 100% clustering with the Cosmopolitan genotype strain. Bleeding from several sites, jaundice, abdominal pain, arthralgia, haemoconcentration, and thrombocytopenia were significantly higher in dengue hemorrhagic cases. However, the rash was significantly more common in Chikungunya patients. Most of the Dengue and Chikungunya positive cases (Age group 6-40 years) were seen in post monsoon season (September to November). Conclusion: Only one-third of total VHF cases are positive for either Dengue/Chikungunya or both. This necessitates the screening of other etiologies capable of causing hemorrhagic manifestations.

Keywords: viral hemorrhagic fever, dengue, chikungunya, zika, India

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Authors: Muhammad Bilal Qadir, Danish Umer, Amir Shahzad

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The use of conductive materials in smart and interactive textiles is gaining significant importance for creating value addition, innovation, and functional product development. These products find their potential applications in health monitoring, military, protection, communication, sensing, monitoring, actuation, fashion, and lifestyles. The materials which are most commonly employed in such type of interactive textile include intrinsically conducting polymers, conductive inks, and metallic coating on textile fabrics and inherently conducting metallic fibre yarns. In this study, silver coated polyester filament yarn is explored for the development of multifunctional interactive gloves. The composite yarn was developed by covering the silver coated polyester filament around the polyester spun yarn using hollow spindle technique. The electrical and tensile properties of the yarn were studied. This novel yarn was used to manufacture a smart glove to explore the antibacterial, functional, and interactive properties of the yarn. The change in electrical resistance due to finger movement at different bending positions and antimicrobial properties were studied. This glove was also found useful as an interactive tool to operate the commonly used touch screen devices due to its conductive nature. The yarn can also be used to develop the sensing elements like stretch, strain, and piezoresistive sensors. Such sensor can be effectively used in medical and sports textile for performance monitoring, vital signs monitoring and development of antibacterial textile for healthcare and hygiene.

Keywords: conductive yarn, interactive textiles, piezoresistive sensors, smart gloves

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Authors: Amir Saber Gharamaleki, Beitollah Alipour, Zeinab Faghfoori, Ahmad YariKhosroushahi

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Colorectal cancer (CRC) is one of the most prevalent cancers and intestinal microbial community plays an important role in colorectal tumorigenesis. Probiotics have recently been assessed as effective anti-proliferative agents and thus this study was performed to examine whether CRC undergo apoptosis by treating with isolated Iranian native dairy yeast, Kluyveromyces marxianus YAS, secretion metabolites. The cytotoxicity assessments on cells (HT-29, Caco-2) were accomplished through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay as well as qualitative DAPI (4',6-diamidino-2-phenylindole staining) and quantitative (flow cytometry assessments) evaluations of apoptosis. To evaluate the main mechanism of apoptosis, Real time PCR method was applied. Kluyveromyces marxianus YAS secretions (IC50) showed significant cytotoxicity against HT-29 and Caco-2 cancer cell lines (66.57 % and 66.34 % apoptosis) similar to 5-Fluorouracil (5-FU) while apoptosis only was developed in 27.57 % of KDR normal cells. The prophylactic effects of Kluyveromyces marxianus (PTCC 5195), as a reference yeast, was not similar to Kluyveromyces marxianus YAS indicating strain dependency of bioactivities on CRC disease prevention. Based on real time PCR results, the main cytotoxicity is related to apoptosis phenomenon and the core related mechanism is depended on the overexpression of BAX, CASP 9, CASP 8 and CASP 3 inducing apoptosis genes. However, several investigations should be conducted to precisely determine the effective compounds to be used as anticancer therapeutics in the future.

Keywords: anticancer, anti-proliferative, apoptosis, cytotoxicity, yeast

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Authors: Hanane Boutaj, Abdelilah Meddich, Said Wahbi, Zainab El Alaoui-Talibi, Allal Douira, Abdelkarim Filali-Maltouf, Cherkaoui El Modafar

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The present work aims to investigate the effect of arbuscular mycorrhizal fungi (AMF) in improving the olive tree resistance to Verticillium wilt caused by Verticillium dahliae. Inoculated plants with a mycorrhizal autochthonous consortium 'Rhizolive consortium' and pure strain 'Glomus irregulare' were infected after three months with V. dahliae. The improving of olive tree resistance was determined through disease severity, incidence, and defoliation. On the other hand, the defense mechanisms of olive plants were evaluated through lignin content, phenylalanine ammonia lyase (PAL) activity, and polyphenol content. The results revealed that both AMF significantly (p < 0.05) reduced disease development and the rate of defoliation in infected olive plants. Moreover, the contents of lignin were boosted after mycorrhizal inoculation in both the roots and the stems of olive plants, which remained significantly (p < 0.001) higher after the 90th days of V. dahliae inoculation. PAL activity was increased after V. dahliae inoculation in the stems of 'Rhizolive consortium' treatment that were 17 times higher than those in the roots of olive plants. The polyphenol content in the stems was about twice higher than those in the roots. The reduction of disease severity was accompanied by increased levels of lignin content, PAL activity, and polyphenol content, particularly in the stems of olive plants, indicating the strengthening of the olive plant immune system against V. dahliae.

Keywords: olive tree, Mycorrhizal autochthonous consortium, Glomus irregulare, Verticillium dahliae, defense mechanisms

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Authors: Afsheen Aman, Shah Ali Ul Qader

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Dextransucrase [EC 2.4.1.5] is a glucosyltransferase that catalysis the biosynthesis of a natural biopolymer called dextran. It can catalyze the transfer of D-glucopyranosyl residues from sucrose to the main chain of dextran. This unique biopolymer has multiple applications in several industries and the key utilization of dextran lies on its molecular weight and the type of branching. Extracellular dextransucrase from Leuconostoc mesenteroides is most extensively studied and characterized. Limited data is available regarding cell-bound or intracellular dextransucrase and on the characterization of dextran produced by in-vitro reaction of intracellular dextransucrase. L. mesenteroides AA1 is reported to produce extracellular dextransucrase that catalyzes biosynthesis of a high molecular weight dextran with only α-(1→6) linkage. Current study deals with the characterization of an intracellular dextransucrase and in vitro biosynthesis of low molecular weight dextran from L. mesenteroides AA1. Intracellular dextransucrase was extracted from cytoplasm and purified to homogeneity for characterization. Kinetic constants, molecular weight and N-terminal sequence analysis of intracellular dextransucrase reveal unique variation with previously reported extracellular dextransucrase from the same strain. In vitro synthesized biopolymer was characterized using NMR spectroscopic techniques. Intracellular dextransucrase exhibited Vmax and Km values of 130.8 DSU ml-1 hr-1 and 221.3 mM, respectively. Optimum catalytic activity was detected at 35°C in 0.15 M citrate phosphate buffer (pH-5.5) in 05 minutes. Molecular mass of purified intracellular dextransucrase is approximately 220.0 kDa on SDS-PAGE. N-terminal sequence of the intracellular enzyme is: GLPGYFGVN that showed no homology with previously reported sequence for the extracellular dextransucrase. This intracellular dextransucrase is capable of in vitro synthesis of dextran under specific conditions. This intracellular dextransucrase is capable of in vitro synthesis of dextran under specific conditions and this biopolymer can be hydrolyzed into different molecular weight fractions for various applications.

Keywords: characterization, dextran, dextransucrase, leuconostoc mesenteroides

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Authors: Malsawmtluanga, J. Malsawma, R. P. Tiwari, V. K. Gahalaut

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North East India is seismically one of the six most active regions of the world. It is placed in Zone V, the highest zone in the seismic zonation of India. It lies at the junction of Himalayan arc to the north and the Burmese arc to the east. The region has witnessed at least 18 large earthquakes including two great earthquakes Shillong (1987, M=8.7) and the Assam Tibet border (1950, M=8.7).The prominent Chite fault lies at the heart of Aizawl, the capital of Mizoram state and this hilly city is the home to about 2 million people. Geologically the area is a part of the Indo-Burmese Wedge and is prone to natural and man-made disasters. Unplanned constructions and urban dwellings on a rapid scale have lead to numerous unsafe structures adversely affecting the ongoing development and welfare projects of the government and they pose a huge threat for earthquakes. Crustal deformation measurements using campaign mode GPS were undertaken across this fault. Campaign mode GPS data were acquired and were processed with GAMIT-GLOBK software. The study presents the current velocity estimates at all the sites in ITRF 2008 and also in the fixed Indian reference frame. The site motion showed that there appears to be no differential motion anywhere across the fault area, thus confirming presently the fault is neither accumulating strain nor slipping aseismically. From the geological and geomorphological evidence, supported by geodetic measurements, lack of historic earthquakes, the Chite fault favours aseismic behaviour in this part of the Indo Burmese Arc (IBA).

Keywords: Chite fault, crustal deformation, geodesy, GPS, IBA

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Authors: Bijay Kumar Sahoo

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An important feature of InₓGa₁-ₓN/GaN heterostructures is strong built-in polarization (BIP) electric field at the hetero-interface due to spontaneous (sp) and piezoelectric (pz) polarizations. The intensity of this electric field reaches several MV/cm. This field has profound impact on optical, electrical and thermal properties. In this work, the effect of BIP field on thermal conductivity of InₓGa₁-ₓN/GaN heterostructure has been investigated theoretically. The interaction between the elastic strain and built in electric field induces additional electric polarization. This additional polarization contributes to the elastic constant of InₓGa₁-ₓN alloy. This in turn modifies material parameters of InₓGa₁-ₓN. The BIP mechanism enhances elastic constant, phonon velocity and Debye temperature and their bowing constants in InₓGa₁-ₓN alloy. These enhanced thermal parameters increase phonon mean free path which boost thermal conduction process. The thermal conductivity (k) of InxGa1-xN alloy has been estimated for x=0, 0.1, 0.3 and 0.9. Computation finds that irrespective of In content, the room temperature k of InₓGa₁-ₓN/GaN heterostructure is enhanced by BIP mechanism. Our analysis shows that at a certain temperature both k with and without BIP show crossover. Below this temperature k with BIP field is lower than k without BIP; however, above this temperature k with BIP field is significantly contributed by BIP mechanism leading to k with BIP field become higher than k without BIP field. The crossover temperature is primary pyroelectric transition temperature. The pyroelectric transition temperature of InₓGa₁-ₓN alloy has been predicted for different x. This signature of pyroelectric nature suggests that thermal conductivity can reveal pyroelectricity in InₓGa₁-ₓN alloy. The composition dependent room temperature k for x=0.1 and 0.3 are in line with prior experimental studies. The result can be used to minimize the self-heating effect in InₓGa₁-ₓN/GaN heterostructures.

Keywords: built-in polarization, phonon relaxation time, thermal properties of InₓGa₁-ₓN /GaN heterostructure, self-heating

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Authors: Mark Linden, Trisha Forbes, Michael Brown, Lynne Marsh, Maria Truesdale, Stuart Todd, Nathan Hughes

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Background The COVID-19 pandemic resulted in a reduction of health care services for many family carers of people with profound and multiple intellectual disabilities (PMID). Due to lack of services, family carers turned to charities for support during the pandemic. We explored the views of charity workers across the UK and Ireland who supported family carers during the COVID-19 pandemic and explored their views on effective online support programmes for family carers. Methods This was a qualitative study using online focus groups with participants (n = 24) from five charities across the UK and Ireland. Questions focused on challenges, supports, coping and resources which helped during lockdown restrictions. Focus groups were audio recorded, transcribed verbatim, and analysed through thematic analysis. Findings Four themes were identified (i) ‘mental and emotional health’, (ii) ‘they who shout the loudest’ (fighting for services), (iii) ‘lack of trust in statutory services’ and (iv) ‘creating an online support programme’. Mental and emotional health emerged as the most prominent theme and included three subthemes named as ‘isolation’, ‘fear of COVID-19’ and ‘the exhaustion of caring’. Conclusions The withdrawal of many services during the COVID-19 pandemic further isolated and placed strain on family carers. Even after the end of the pandemic family cares continue to report on the struggle to receive adequate support. There is a critical need to design services, including online support programmes, in partnership with family carers which adequately address their needs.

Keywords: intellectual disability, family carers, COVID-19, charities

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Authors: Shahriar Sepahvand, Mohammad Ali Davarpanah

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Infections caused by Acinetobacter baumannii are among the common hospital-acquired infections that have seen an increase in antibiotic resistance in recent years. Colistin is the last treatment option against this pathogen. The aim of this study is to investigate the histopathology of BALB/C mice receiving sensitive and resistant strains of Acinetobacter baumannii to colistin. A total of 68 female laboratory mice weighing 30 to 40 grams of the BALB/C breed were studied in this research for three weeks under appropriate laboratory conditions in terms of food and environment. The experimental groups included: control group, second group, third group, fourth group. Lung, liver, spleen, and kidney tissues were removed from anesthetized mice and, after washing in physiological serum, were fixed in 10% formalin for 14 days. For dehydration, alcohol with ascending degrees of 70, 80, 90, and 100 was used. After clearing and soaking in paraffin, the samples were embedded in paraffin. Then, sections with a thickness of 5 microns were prepared and, after staining by hematoxylin-eosin, the samples were ready for study with a light microscope. In liver, spleen, lung, and kidney tissues of mice receiving the colistin-sensitive strain of Acinetobacter baumannii, infiltration of inflammatory cells and hyperemia were observed compared to control group mice. Liver and lung tissues of mice receiving strains of Acinetobacter baumannii resistant to colistin showed tissue destruction in addition to infiltration of inflammatory cells and hyperemia, with more destruction observed in lung tissue.

Keywords: acinetobacter baumannii, colistin antibiotic, histopathological examination, resistant

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Authors: Asavasereerat K., Teacharsripaitoon T., Tungyingyong P., Charupongrat S., Noppiboon S. Hochareon L., Kitsuban P.

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Third generation of vaccines is based on gene therapy where DNA is introduced into patients. The antigenic or therapeutic proteins encoded from transgenes DNA triggers an immune-response to counteract various diseases. Moreover, DNA vaccine offers the customization of its ability on protection and treatment with high stability. The production of DNA vaccines become of interest. According to USFDA guidance for industry, the recommended limits for impurities from host cell are lower than 1%, and the active conformation homogeneity supercoiled DNA, is more than 80%. Thus, the purification strategy using two-steps chromatography has been established and verified for its robustness. Herein, pVax1/lacZ, a pre-approved USFDA DNA vaccine backbone, was used and transformed into E. coli strain DH5α. Three purification process parameters including sample-loading flow rate, the salt concentration in washing and eluting buffer, were studied and the experiment was designed using response surface method with central composite face-centered (CCF) as a model. The designed range of selected parameters was 10% variation from the optimized set point as a safety factor. The purity in the percentage of supercoiled conformation obtained from each chromatography step, AIEX and HIC, were analyzed by HPLC. The response data were used to establish regression model and statistically analyzed followed by Monte Carlo simulation using SAS JMP. The results on the purity of the product obtained from AIEX and HIC are between 89.4 to 92.5% and 88.3 to 100.0%, respectively. Monte Carlo simulation showed that the pVAX1/lacZ purification process is robust with confidence intervals of 0.90 in range of 90.18-91.00% and 95.88-100.00%, for AIEX and HIC respectively.

Keywords: AIEX, DNA vaccine, HIC, puification, response surface method, robustness

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Authors: Cristina Busuioc, Georgeta Voicu, Sorin-Ion Jinga

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The human bone presents in its dry form piezoelectric properties, which means that a mechanical stress results in electric polarization and an applied electric field causes strain. The immediate consequence was the revealing of piezoelectricity role in bone remodelling, as well as the integration of ceramic materials with piezoelectric behaviour in the composition of unitary or composite biomaterials. Thus, we prepared hydroxyapatite - collagen hybrid materials with barium titanate addition in order to achieve a better osseointegration. Barium titanate powder synthesized by a combined sol-gel-hydrothermal method, commercial hydroxyapatite and laboratory extracted collagen gel were employed as starting materials. Before the composites, fabrication, the powder with piezoelectric features was characterized in detail from the compositional, structural, morphological and electrical point of view. The next step was to elucidate the influence of barium titanate presence especially on the biological properties of the final materials. The biocompatibility of the hybrid supports without or with piezoelectric addition was investigated on mouse osteoblast cells through LDH cytotoxicity assay, LIVE/DEAD cell viability assay, and MTT cell proliferation assay. All results indicated that the analysed materials do not exert cytotoxic effects and present the ability to sustain cell survival and to promote their proliferation. In conclusion, barium titanate nanoparticles exhibit a good biocompatibility and osteoinductive properties, while the derived composite materials based on hydroxyapatite as oxide phase and collagen as polymeric phase can be successfully used for tissue engineering applications.

Keywords: barium titanate, hybrid composites, piezoelectricity, tissue engineering

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Authors: Sadegh Lotfieblisofla, Arash Khodabakhshi

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Tissue plasminogen activator (tPA) as a serine protease plays an important role in the fibrinolytic system and the dissolution of fibrin clots in human body. The production of this drug in plants such as tobacco could reduce its production costs. In this study, expression of tPA gene and protein targeting to different plant cell compartments, using various signal peptides has been investigated. For high level of expression, Kozak sequence was used after CaMV35S in the beginning of the gene. In order to design the final construction, Extensin, KDEL (amino acid sequence including Lys-Asp-Glu-Leu) and SP (γ-zein signal peptide coding sequence) were used as leader signals to conduct this protein into apoplast, endoplasmic reticulum and cytosol spaces, respectively. Cloned human tPA gene under the CaMV (Cauliflower mosaic virus) 35S promoter and NOS (Nopaline Synthase) terminator into pBI121 plasmid was transferred into tobacco explants by Agrobacterium tumefaciens strain LBA₄₄₀₄. The presence and copy number of genes in transgenic tobacco was proved by Southern blotting. Enzymatic activity of the rt-PA protein in transgenic plants compared to non-transgenic plants was confirmed by Zymography assay. The presence and amount of rt-PA recombinant protein in plants was estimated by ELISA analysis on crude protein extract of transgenic tobacco using a specific antibody. The yield of recombinant tPA in transgenic tobacco for SP, KDEL, Extensin signals were counted 0.50, 0.68, 0.69 microgram per milligram of total soluble proteins.

Keywords: tPA, recombinant, transgenic, tobacco

Procedia PDF Downloads 127

Authors: Javad Saeidaskari, Nader Khalafian

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The undesirable impact of vibrations induced by road and railway traffic is an important concern in modern world. These vibrations are transmitted through soil and cause disturbances to the residence area and high-tech production facilities alongside the train/traffic lines. In this paper for the first time a new method of soil improvement with vibration absorber material, is used to increase the damping factor, in other word, to reduce the ability of wave transitions in sand. In this study standard Firoozkooh No. 161 sand is used as the host sand. The semi rigid polyurethane (PU) foam which used in this research is one of the common materials for vibration absorbing purposes. Series of cyclic triaxial tests were conducted on remolded samples with identical relative density of 70% of maximum dry density for different volume percentage of shredded PU foam. The frequency of tests was 0.1 Htz with shear strain of 0.37% and 0.75% and also the effective confining pressures during the tests were 100 kPa and 350 kPa. In order to find out the best soil-PU foam mixture, different volume percent of PU foam varying from 10% to 30% were examined. The results show that adding PU foam up to 20%, as its optimum content, causes notable enhancement in damping ratio for both shear strains of 0.37% (52.19% and 69% increase for effective confining pressures of 100 kPa and 350 kPa, respectively) and 0.75% (59.56% and 59.11% increase for effective confining pressures of 100 kPa and 350 kPa, respectively). The results related to shear modulus present significant reduction for both shear strains of 0.37% (82.22% and 56.03% decrease for effective confining pressures of 100 kPa and 350 kPa, respectively) and 0.75% (89.32% and 39.9% decrease for effective confining pressures of 100 kPa and 350 kPa, respectively). In conclusion, shredded PU foams effectively affect the dynamic properties of sand and act as vibration absorber in soil.

Keywords: polyurethane foam, sand, damping ratio, shear modulus

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Authors: Aid Abdelkrim

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A method of fatigue damage accumulation based upon application of energy parameters of the fatigue process is proposed in the paper. Using this model is simple, it has no parameter to be determined, it requires only the knowledge of the curve W–N (W: strain energy density N: number of cycles at failure) determined from the experimental Wöhler curve. To examine the performance of nonlinear models proposed in the estimation of fatigue damage and fatigue life of components under random loading, a batch of specimens made of 6082 T 6 aluminium alloy has been studied and some of the results are reported in the present paper. The paper describes an algorithm and suggests a fatigue cumulative damage model, especially when random loading is considered. This work contains the results of uni-axial random load fatigue tests with different mean and amplitude values performed on 6082T6 aluminium alloy specimens. The proposed model has been formulated to take into account the damage evolution at different load levels and it allows the effect of the loading sequence to be included by means of a recurrence formula derived for multilevel loading, considering complex load sequences. It is concluded that a ‘damaged stress interaction damage rule’ proposed here allows a better fatigue damage prediction than the widely used Palmgren–Miner rule, and a formula derived in random fatigue could be used to predict the fatigue damage and fatigue lifetime very easily. The results obtained by the model are compared with the experimental results and those calculated by the most fatigue damage model used in fatigue (Miner’s model). The comparison shows that the proposed model, presents a good estimation of the experimental results. Moreover, the error is minimized in comparison to the Miner’s model.

Keywords: damage accumulation, energy model, damage indicator, variable loading, random loading

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Authors: A. A. M. Azoddein, Y. Nuratri, A. B. Bustary, F. A. M. Azli, S. C. Sayuti

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Pseudomonas putida is a potential strain in biological treatment to remove mercury contained in the effluent of petrochemical industry due to its mercury reductase enzyme that able to reduce ionic mercury to elementary mercury. Freeze-dried P. putida allows easy, inexpensive shipping, handling and high stability of the product. This study was aimed to freeze dry P. putida cells with addition of lyoprotectant. Lyoprotectant was added into the cells suspension prior to freezing. Dried P. putida obtained was then mixed with synthetic mercury. Viability of recovery P. putida after freeze dry was significantly influenced by the type of lyoprotectant. Among the lyoprotectants, tween 80/ sucrose was found to be the best lyoprotectant. Sucrose able to recover more than 78% (6.2E+09 CFU/ml) of the original cells (7.90E+09CFU/ml) after freeze dry and able to retain 5.40E+05 viable cells after 4 weeks storage in 4oC without vacuum. Polyethylene glycol (PEG) pre-treated freeze dry cells and broth pre-treated freeze dry cells after freeze-dry recovered more than 64% (5.0 E+09CFU/ml) and >0.1% (5.60E+07CFU/ml). Freeze-dried P. putida cells in PEG and broth cannot survive after 4 weeks storage. Freeze dry also does not really change the pattern of growth P. putida but extension of lag time was found 1 hour after 3 weeks of storage. Additional time was required for freeze-dried P. putida cells to recover before introduce freeze-dried cells to more complicated condition such as mercury solution. The maximum mercury reduction of PEG pre-treated freeze-dried cells after freeze dry and after storage 3 weeks was 56.78% and 17.91%. The maximum of mercury reduction of tween 80/sucrose pre-treated freeze-dried cells after freeze dry and after storage 3 weeks were 26.35% and 25.03%. Freeze dried P. putida was found to have lower mercury reduction compare to the fresh P. putida that has been growth in agar. Result from this study may be beneficial and useful as initial reference before commercialize freeze-dried P. putida.

Keywords: Pseudomonas putida, freeze-dry, PEG, tween80/Sucrose, mercury, cell viability

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Authors: Manendra Singh Parihar, Sandip Ghosh Chowdhury

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Austenitic stainless steels are widely used and give a good combination of properties. When this steel is plastically deformed, a phase transformation of the metastable Face Centred Cubic Austenite to the stable Body Centred Cubic (α’) or to the Hexagonal close packed (ԑ) martensite may occur, leading to the enhancement in the mechanical properties like strength. The work was based on variant selection and corresponding texture analysis for the strain induced martensitic transformation during deformation of the parent austenite FCC phase to form the product HCP and the BCC martensite phases separately, obeying their respective orientation relationships. The automated method for reconstruction of the parent phase orientation using the EBSD data of the product phase orientation is done using the MATLAB and TSL-OIM software. The method of triplets was used which involves the formation of a triplet of neighboring product grains having a common variant and linking them using a misorientation-based criterion. This led to the proper reconstruction of the pre-transformation phase orientation data and thus to its micro structure and texture. The computational speed of current method is better compared to the previously used methods of reconstruction. The reconstruction of austenite from ԑ and α’ martensite was carried out for multiple samples and their IPF images, pole figures, inverse pole figures and ODFs were compared. Similar type of results was observed for all samples. The comparison gives the idea for estimating the correct sequence of the transformation i.e. γ → ε → α’ or γ → α’, during deformation of AISI 304 austenitic stainless steel.

Keywords: variant selection, reconstruction, EBSD, austenitic stainless steel, martensitic transformation

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Authors: Sidra Shaheen, Nadir Naveed Siddiqui, Shah Ali Ul Qader

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In recent years, significant progress has been made in discovering and developing new bacterial polysaccharides producing organisms possessing extremely functional properties. Levan is a natural biopolymer of fructose which is produced by transfructosylation reaction in the presence of levansucrase. It is one of the industrially promising enzymes that offer a variety of industrial applications in the field of cosmetics, foods and pharmaceuticals. Although levan has significant applications but the yield of levan produced is not equal to other biopolymers due to the inefficiency of producer microorganism. Among wide range of levansucrase producing microorganisms, Zymomonas mobilis is considered as a potential candidate for large scale production of this natural polysaccharide. The present investigation is concerned with the isolation of levansucrase producing natural isolate having maximum enzyme production. Furthermore, production parameters were optimized to get higher enzyme yield. Levansucrase was partially purified and characterized to study its applicability on industrial scale. The results of this study revealed that the bacterial strain Z. mobilis KIBGE-IB14 was the best producer of levansucrase. Bacterial growth and enzyme production was greatly influenced by physical and chemical parameters. Maximum levansucrase production was achieved after 24 hours of fermentation at 30°C using modified medium of pH-6.5. Contrary to other levansucrases, the one presented in the current study is able to produce high amount of products in relatively short period of time with optimum temperature at 35°C. Due to these advantages, this enzyme can be used on large scale for commercial production of levan and other important metabolites.

Keywords: levansucrase, metabolites, polysaccharides, transfructosylation

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Authors: M. L. Burnwal, P. Raychowdhury

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Earthquake leads to adverse effects on buildings resting on soft soils. Mitigating the response of shallow foundations on soft soil with different methods reduces settlement and provides foundation stability. Few methods such as the rocking foundation (used in Performance-based design), deep foundation, prefabricated drain, grouting, and Vibro-compaction are used to control the pore pressure and enhance the strength of the loose soils. One of the problems with these methods is that the settlement is uncontrollable, leading to differential settlement of the footings, further leading to the collapse of buildings. The present study investigates the utility of geosynthetics as a potential improvement of the subsoil to reduce the earthquake-induced settlement of structures. A steel moment-resisting frame building resting on loose liquefiable dry soil, subjected to Uttarkashi 1991 and Chamba 1995 earthquakes, is used for the soil-structure interaction (SSI) analysis. The continuum model can simultaneously simulate structure, soil, interfaces, and geogrids in the OpenSees framework. Soil is modeled with PressureDependentMultiYield (PDMY) material models with Quad element that provides stress-strain at gauss points and is calibrated to predict the behavior of Ganga sand. The model analyzed with a tied degree of freedom contact reveals that the system responses align with the shake table experimental results. An attempt is made to study the responses of footing structure and geosynthetics with unreinforced and reinforced bases with varying parameters. The result shows that geogrid reinforces shallow foundation effectively reduces the settlement by 60%.

Keywords: settlement, shallow foundation, SSI, continuum FEM

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Authors: Niloofar Bahramian, Mohammad Atai, Mohammad Reza Naimi-Jamal

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Rule of mixtures is the simple analytical model is used to predict various properties of composites before design. The aim of this study was to demonstrate the benefits and limitations of the Rule-of-Mixtures (ROM) for predicting bending modulus of a continuous and unidirectional fiber reinforced composites using in dental applications. The Composites were fabricated from light curing resin (with and without silica nanoparticles) and modified and non-modified fibers. Composite samples were divided into eight groups with ten specimens for each group. The bending modulus (flexural modulus) of samples was determined from the slope of the initial linear region of stress-strain curve on 2mm×2mm×25mm specimens with different designs: fibers corona treatment time (0s, 5s, 7s), fibers silane treatment (0%wt, 2%wt), fibers volume fraction (41%, 33%, 25%) and nanoparticles incorporation in resin (0%wt, 10%wt, 15%wt). To study the fiber and matrix interface after fracture, single edge notch beam (SENB) method and scanning electron microscope (SEM) were used. SEM also was used to show the nanoparticles dispersion in resin. Experimental results of bending modulus for composites made of both physical (corona) and chemical (silane) treated fibers were in reasonable agreement with linear ROM estimates, but untreated fibers or non-optimized treated fibers and poor nanoparticles dispersion did not correlate as well with ROM results. This study shows that the ROM is useful to predict the mechanical behavior of unidirectional dental composites but fiber-resin interface and quality of nanoparticles dispersion play important role in ROM accurate predictions.

Keywords: bending modulus, fiber reinforced composite, fiber treatment, rule-of-mixtures

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Authors: Arash Sadeghi, Taimaz Larimian

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This study explores the intricate interplay between community-based social capital (CBSC) and neighbourhood dynamics in enhancing resilience of Iranian SMEs, particularly under the strain of international sanctions. Utilising fuzzy-set Qualitative Comparative Analysis (fsQCA), we examine how different dimensions of CBSC—structural, relational, and cognitive—interact with neighbourhood socio-economic and built-environment characteristics to influence SME resilience. Findings reveal four configurations that contribute to the presence of resistance and five configurations associated with the adaptation outcome. Each configuration demonstrates a distinct combination of social capital elements, which vary according to the specific socio-economic and built-environmental characteristics of the neighbourhoods. The first configuration highlights the importance of structural social capital in deprived areas for building resistance, while the second emphasises the role of relational social capital in low-density, minimally deprived areas. Overall, cognitive social capital seems to be less effective in driving economic resilience compared to structural and relational types. This research contributes to the literature by providing a nuanced understanding of the synergistic effects of CBSC dimensions and neighbourhood characteristics on SME resilience. By adopting a configurational approach, we move beyond traditional methodologies, offering a comprehensive view of the complex dynamics of CBSC and neighbourhood characteristics and their impact on SME resilience in varying neighbourhoods.

Keywords: community-based social capital, fuzzy-set qualitative comparative analysis (fsQCA), place-based resilience, resistance

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Authors: Gamal Seliem, Hassan Seliem

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Aswan area is one of the important areas in Egypt and because it encompasses the vital engineering structure of the High dam, so it has been selected for the present study. The study of the crustal deformation and gravity associated with earthquake activity in the High Dam area of great importance for the safety of the High Dam and its economic resources. This paper deals with using micro-gravity, precise leveling and GPS data for geophysical and geodetically studies. For carrying out the detailed gravity survey in the area, were established for studying the subsurface structures. To study the recent vertical movements, a profile of 10 km length joins the High Dam and Aswan old dam were established along the road connecting the two dams. This profile consists of 35 GPS/leveling stations extending along the two sides of the road and on the High Dam body. Precise leveling was carried out with GPS and repeated micro-gravity survey in the same time. GPS network consisting of nine stations was established for studying the recent crustal movements. Many campaigns from December 2001 to December 2014 were performed for collecting the gravity, leveling and GPS data. The main aim of this work is to study the structural features and the behavior of the area, as depicted from repeated micro-gravity, precise leveling and GPS measurements. The present work focuses on the analysis of the gravity, leveling and GPS data. The gravity results of the present study investigate and analyze the subsurface geologic structures and reveal to there be minor structures; features and anomalies are taking W-E and N-S directions. The geodetic results indicated lower rates of the vertical and horizontal displacements and strain values. This may be related to the stability of the area.

Keywords: repeated micro-gravity changes, precise leveling, GPS data, Aswan High Dam

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Authors: Anyaphat Srithanasuwan, Noppason Pangprasit, Montira Intanon, Phongsakorn Chuammitri, Witaya Suriyasathaporn, Ynte H. Schukken

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Streptococcus uberis is one of the most common mastitis-causing pathogens, with a wide range of intramammary infection (IMI) durations and pathogenicity. This study aimed to compare shared or unique virulence factor gene clusters distinguishing persistent and transient strains of S. uberis. A total of 139 S. uberis strains were isolated from three small-holder dairy herds with a high prevalence of S. uberis mastitis. The duration of IMI was used to categorize bacteria into two groups: transient and persistent strains with an IMI duration of less than 1 month and longer than 2 months, respectively. Six representative S. uberis strains, three from each group (transience and persistence) were selected for analysis. All transient strains exhibited multi-locus sequence types (MLST), indicating a highly diverse population of transient S. uberis. In contrast, MLST of persistent strains was available in an online database (pubMLST). Identification of virulence genes was performed using whole-genome sequencing (WGS) data. Differences in genomic size and number of virulent genes were found. For example, the BCA gene or alpha-c protein and the gene associated with capsule formation (hasAB), found in persistent strains, are important for attachment and invasion, as well as the evasion of the antimicrobial mechanisms and survival persistence, respectively. These findings suggest a genetic-level difference between the two strain types. Consequently, a comprehensive study of 139 S. uberis isolates will be conducted to perform an in-depth genetic assessment through WGS analysis on an Illumina platform.

Keywords: Streptococcus Uberis, mastitis, whole genome sequence, intramammary infection, persistent S. Uberis, transient s. Uberis

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Authors: Jae-Yong Park, Gwancheol Seo, Young-Ho Kim

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Microvoids, sometimes called Kikendall voids, generally form in the interfaces between Sn-based solders and Cu and degrade the mechanical and electrical properties of the solder joints. The microvoid formation is known as the rapid interdiffusion between Sn and Cu and impurity content in the Cu. Cu electroplating from the acid solutions has been widely used by microelectronic packaging industry for both printed circuit board (PCB) and integrated circuit (IC) applications. The quality of electroplated Cu that can be optimized by the electroplating conditions is critical for the solder joint reliability. In this paper, the influence of electroplating conditions on the microvoid growth in the interfaces between Sn-3.0Ag-0.5Cu (SAC) solder and Cu layer was investigated during isothermal aging. The Cu layers were electroplated by controlling the additive of electroplating bath and current density to induce various microvoid densities. The electroplating bath consisted of sulfate, sulfuric acid, and additives and the current density of 5-15 mA/cm2 for each bath was used. After aging at 180 °C for up to 250 h, typical bi-layer of Cu6Sn5 and Cu3Sn intermetallic compounds (IMCs) was gradually growth at the SAC/Cu interface and microvoid density in the Cu3Sn showed disparities in the electroplating conditions. As the current density increased, the microvoid formation was accelerated in all electroplating baths. The higher current density induced, the higher impurity content in the electroplated Cu. When the polyethylene glycol (PEG) and Cl- ion were mixed in an electroplating bath, the microvoid formation was the highest compared to other electroplating baths. On the other hand, the overall IMC thickness was similar in all samples irrespective of the electroplating conditions. Impurity content in electroplated Cu influenced the microvoid growth, but the IMC growth was not affected by the impurity content. In conclusion, the electroplated conditions are properly optimized to avoid the excessive microvoid formation that results in brittle fracture of solder joint under high strain rate loading.

Keywords: electroplating, additive, microvoid, intermetallic compound

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Authors: Kevin Giraldo, Juan A. Gallego, Uriel Zapata, Fanny L. Casado

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Compliant mechanisms are designed to deform in a controlled manner in response to external forces, utilizing the flexibility of their components to store potential elastic energy during deformation, gradually releasing it upon returning to its original form. This article explores the design of a knee orthosis intended to assist users during stand-up motion. The orthosis makes use of a compliant mechanism to balance the user’s weight, thereby minimizing the strain on leg muscles during standup motion. The primary function of the compliant mechanism is to store and exchange potential energy, so when coupled with the gravitational potential of the user, the total potential energy variation is minimized. The design process for the semi-rigid knee orthosis involved material selection and the development of a numerical model for the compliant mechanism seen as a spring. Geometric properties are obtained through the numerical modeling of the spring once the desired stiffness and safety factor values have been attained. Subsequently, a 3D finite element analysis was conducted. The study demonstrates a strong correlation between the maximum stress in the mathematical model (250.22 MPa) and the simulation (239.8 MPa), with a 4.16% error. Both analyses safety factors: 1.02 for the mathematical approach and 1.1 for the simulation, with a consistent 7.84% margin of error. The spring’s stiffness, calculated at 90.82 Nm/rad analytically and 85.71 Nm/rad in the simulation, exhibits a 5.62% difference. These results suggest significant potential for the proposed device in assisting patients with knee orthopedic restrictions, contributing to ongoing efforts in advancing the understanding and treatment of knee osteoarthritis.

Keywords: biomechanics, complaint mechanisms, gonarthrosis, orthoses.

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Authors: Maia Kukhaleishvili, Ekaterine Bulauri, Iveta Megrelishvili, Tamar Shamatava, Tamar Chipashvili

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Plant viruses can cause loss of yield and quality in a lot of important crops. Symptoms of pathogens are variable depending on the cultivars and virus strain. Selection of resistant potato varieties would reduce the risk of virus transmission and significant economic impact. Other way to avoid reduced harvest yields is regular potato seed production sampling and testing for viral infection. The aim of this study was to determine the occurrence and distribution of viral diseases according potato cultivars for further selection of virus-free material in Georgia. During the summer 2015- 2016, 5 potato cultivars (Sante, Laura, Jelly, Red Sonia, Anushka) at 5 different farms located in Akhalkalaki were tested for 6 different potato viruses: Potato virus A (PVA), Potato virus M (PVM), Potato virus S (PVS), Potato virus X (PVX), Potato virus Y (PVY) and potato leaf roll virus (PLRV). A serological method, Double Antibody Sandwich-Enzyme linked Immunosorbent Assay (DASELISA) was used at the laboratory to analyze the results. The result showed that PVY (21.4%) and PLRV (19.7%) virus presence in collected samples was relatively high compared to others. Researched potato cultivars except Jelly and Laura were infected by PVY with different concentrations. PLRV was found only in three potato cultivars (Sante, Jelly, Red Sonia) and PVM virus (3.12%) was characterized with low prevalence. PVX, PVA and PVS virus infection was not reported. It would be noted that 7.9% of samples were containing PVY/PLRV mix infection. Based on the results it can be concluded that PVY and PLRV infections are dominant in all research cultivars. Therefore significant yield losses are expected. Systematic, long-term control of potato viral infection, especially seed-potatoes, must be regarded as the most important factor to increase seed productivity.

Keywords: virus, potato, infection, diseases

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Authors: Shagufta Gulraiz, Abu Saeed Hashmi, Muhammad Mohsin Javed

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Lysine required for poultry feed is imported in Pakistan to fulfil the desired dietary needs. Present study was designed to produce maximum lysine by utilizing cheap sources to save the foreign exchange. To achieve the goal of lysine production through fermentation, large scale production of lysine was carried out in 7.5 L stirred glass vessel fermenter with wild and mutant Brevibacterium flavum (B. flavum) using all pre-optimized conditions. The identification of produced lysine was carried out by TLC and amino acid analyzer. Toxicity evaluation of produced lysine was performed before feeding to broiler chicks. During biological trial concentrated fermented broth having 8% lysine was used in poultry rations as a source of Lysine for test birds. Fermenter scale studies showed that the maximum lysine (20.8 g/L) was produced at 250 rpm, 1.5 vvm aeration, 6.0% inoculum under controlled pH conditions after 56 h of fermentation with wild culture but mutant (BFENU2) gave maximum yield of lysine 36.3 g/L under optimized condition after 48 h. Amino acid profiling showed 1.826% Lysine in fermented broth by wild B. flavum and 2.644% by mutant strain (BFENU2). Toxicity evaluation report showed that the produced lysine is safe for consumption by broilers. Biological evaluation results showed that produced lysine was equally good as commercial lysine in terms of weight gain, feed intake and feed conversion ratio. A cheap and practical bioprocess of Lysine production was concluded, that can be exploited commercially in Pakistan to save foreign exchange.

Keywords: lysine, fermentation, broiler chicks, biological evaluation

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Authors: Nickolay Nosik, Dmitry Nosik, Marina Bochkova, Nina Kondrashina, Olga Lobach

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The bactericidal effect of UV radiation is known for long time and widely used for inactivation of pathogens but for viruses it is not so uniform. Due to a wide variety of viruses their sensitivity to UV radiation is quite different and not quite predictable. The goal of the study was to determine the inactivation kinetics of UV radiation ( 254 nm) of the viruses of social importance (HIV), as well as test-viruses (poliovirus, adenovirus) used for the evaluation of the viral inactivation efficacy of germicides. Methods: DNA viruses- adenovirus, type 5; Herpes simplex virus (HSV), type 1, and RNA viruses–human immunodeficiency virus (HIV), type 1 and poliovirus, type 1 (Sabin strain) were obtained from State collection of viruses ( The D.I. Ivanovsky Institute of Virology). The source of UV radiation was a 15-watt low-pressure mercury vapor lamp (over 60% 254nm). The samples of 5cm2 were placed direct under the UV lamp flow (h-0.3m). Log reduction value was used as a marker for the rate of virus inactivation. Results: The data obtained indicate that poliovirus (one of the viruses most resistant to chemical germicides) and HSV are rather sensitive to UV radiation ( D90 =250-311 J/m2). Adenovirus is much more resistant to UV radiation (750 J/m2 ). The kinetics of adenovirus inactivation : 0 min- 5.0 lg TCID50, 10 min - 5,0, 15 min -4,0, 30 min – 3.5, 60 min – 1,0, 75 min -0,5 lg TCID50, 90 min –virus not detectable. HIV is most resistant to UV radiation among the studied viruses. It takes more than 4 hrs to inactivate the virus on the surface. D90 = 2000 J/m2 Conclusion: The results of the study show that there is no direct dependence between sensitivity to UV light and the size of the virion or presence\absence of the envelope of the virus. Poliovirus and adenovirus are small viruses (20-30nm poliovirus and 70-90nm adenovirus) and both are non-enveloped viruses but adenovirus 3-fold more resistant to UV radiation than poliovirus. It can be expected that viruses with more complicate structure, like Herpes virus (200nm) or HIV (80-100 nm), would be more sensitive to UV light. However, the very high resistance of HIV to UV radiation needs further investigation. The diverse resistance of the different viruses to UV radiation should be taken into the account when UV light is used to inactivate infectious viruses in hospitals and other public environments.

Keywords: HIV, HSV, inhibition of viruses, UV radiation

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Authors: Anuj Tyagi, Balwinder Singh, Naveen Kumar B. T., Niraj K. Singh

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Characterization of diverse microbial communities in specific environment plays a crucial role in the better understanding of their functional relationship with the ecosystem. It is now well established that gut microbiome of fish is not the simple replication of microbiota of surrounding local habitat, and extensive species, dietary, physiological and metabolic variations in fishes may have a significant impact on its composition. Moreover, overuse of antibiotics in human, veterinary and aquaculture medicine has led to rapid emergence and propagation of antibiotic resistance genes (ARGs) in the aquatic environment. Microbial communities harboring specific ARGs not only get a preferential edge during selective antibiotic exposure but also possess the significant risk of ARGs transfer to other non-resistance bacteria within the confined environments. This phenomenon may lead to the emergence of habitat-specific microbial resistomes and subsequent emergence of virulent antibiotic-resistant pathogens with severe fish and consumer health consequences. In this study, gut microbiota of freshwater carp (Labeo rohita) was investigated by shotgun metagenomics to understand its taxonomic composition and functional capabilities. Metagenomic DNA, extracted from the fish gut, was subjected to sequencing on Illumina NextSeq to generate paired-end (PE) 2 x 150 bp sequencing reads. After the QC of raw sequencing data by Trimmomatic, taxonomic analysis by Kraken2 taxonomic sequence classification system revealed the presence of 36 phyla, 326 families and 985 genera in the fish gut microbiome. At phylum level, Proteobacteria accounted for more than three-fourths of total bacterial populations followed by Actinobacteria (14%) and Cyanobacteria (3%). Commonly used probiotic bacteria (Bacillus, Lactobacillus, Streptococcus, and Lactococcus) were found to be very less prevalent in fish gut. After sequencing data assembly by MEGAHIT v1.1.2 assembler and PROKKA automated analysis pipeline, pathway analysis revealed the presence of 1,608 Metacyc pathways in the fish gut microbiome. Biosynthesis pathways were found to be the most dominant (51%) followed by degradation (39%), energy-metabolism (4%) and fermentation (2%). Almost one-third (33%) of biosynthesis pathways were involved in the synthesis of secondary metabolites. Metabolic pathways for the biosynthesis of 35 antibiotic types were also present, and these accounted for 5% of overall metabolic pathways in the fish gut microbiome. Fifty-one different types of antibiotic resistance genes (ARGs) belonging to 15 antimicrobial resistance (AMR) gene families and conferring resistance against 24 antibiotic types were detected in fish gut. More than 90% ARGs in fish gut microbiome were against beta-lactams (penicillins, cephalosporins, penems, and monobactams). Resistance against tetracycline, macrolides, fluoroquinolones, and phenicols ranged from 0.7% to 1.3%. Some of the ARGs for multi-drug resistance were also found to be located on sequences of plasmid origin. The presence of pathogenic bacteria and ARGs on plasmid sequences suggested the potential risk due to horizontal gene transfer in the confined gut environment.

Keywords: antibiotic resistance, fish gut, metabolic pathways, microbial diversity

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Authors: J. Gabrielczyk, J. Kluitmann, T. Dammeyer, H. J. Jördening

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High-level expression of proteins in bacteria often causes production of insoluble protein aggregates, called inclusion bodies (IB). They contain mainly one type of protein and offer an easy and efficient way to get purified protein. On the other hand, proteins in IB are normally devoid of function and therefore need a special treatment to become active. Most refolding techniques aim at diluting the solubilizing chaotropic agents. Unfortunately, optimal refolding conditions have to be found empirically for every protein. For large-scale applications, a simple refolding process with high yields and high final enzyme concentrations is still missing. The constructed plasmid pASK-IBA63b containing the sequence of fructosyltransferase (FTF, EC 2.4.1.162) from Bacillus subtilis NCIMB 11871 was transformed into E. coli BL21 (DE3) Rosetta. The bacterium was cultivated in a fed-batch bioreactor. The produced FTF was obtained mainly as IB. For refolding experiments, five different amounts of IBs were solubilized in urea buffer with protein concentration of 0.2-8.5 g/L. Solubilizates were refolded with batch or continuous dialysis. The refolding yield was determined by measuring the protein concentration of the clear supernatant before and after the dialysis. Particle size was measured by dynamic light scattering. We tested the solubilization properties of fructosyltransferase IBs. The particle size measurements revealed that the solubilization of the aggregates is achieved at urea concentration of 5M or higher and confirmed by absorption spectroscopy. All results confirm previous investigations that refolding yields are dependent upon initial protein concentration. In batch dialysis, the yields dropped from 67% to 12% and 72% to 19% for continuous dialysis, in relation to initial concentrations from 0.2 to 8.5 g/L. Often used additives such as sucrose and glycerol had no effect on refolding yields. Buffer screening indicated a significant increase in activity but also temperature stability of FTF with citrate/phosphate buffer. By adding citrate to the dialysis buffer, we were able to increase the refolding yields to 82-47% in batch and 90-74% in the continuous process. Further experiments showed that in general, higher ionic strength of buffers had major impact on refolding yields; doubling the buffer concentration increased the yields up to threefold. Finally, we achieved corresponding high refolding yields by reducing the chamber volume by 75% and the amount of buffer needed. The refolded enzyme had an optimal activity of 12.5±0.3 x104 units/g. However, detailed experiments with native FTF revealed a reaggregation of the molecules and loss in specific activity depending on the enzyme concentration and particle size. For that reason, we actually focus on developing a process of simultaneous enzyme refolding and immobilization. The results of this study show a new approach in finding optimal refolding conditions for inclusion bodies at high concentrations. Straightforward buffer screening and increase of the ionic strength can optimize the refolding yield of the target protein by 400%. Gentle removal of chaotrope with continuous dialysis increases the yields by an additional 65%, independent of the refolding buffer applied. In general time is the crucial parameter for successful refolding of solubilized proteins.

Keywords: dialysis, inclusion body, refolding, solubilization

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Authors: Mark Linden, Michael Brown, Lynne Marsh, Maria Truesdale, Stuart Todd, Nathan Hughes, Trisha Forbes, Rachel Leonard

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Background: The COVID-19 pandemic exacerbated the already significant strain placed on family carers of people with profound and multiple intellectual disabilities (PMID), given the withdrawal of many services during lockdown. The aim of this study was to explore the experiences of family carers of people with PMID during the COVID-19 pandemic. Methods: Online focus groups were conducted with family carers (n=126) from across the UK and the Republic of Ireland. Participants were asked about their experiences of the COVID-19 pandemic, coping strategies, and challenges faced. Focus groups were audio recorded, transcribed verbatim and analyzed through thematic analysis. Findings: Three themes emerged from our analysis of the data: (i) COVID-19 as a double-edged sword, (ii) The struggle for support (iii) the Constant nature of caring. These included 11 subthemes: (i) ‘COVID-19 as a catalyst for change’, ‘Challenges during COVID-19: dealing with change’, ‘Challenges during COVID-19: fear of COVID-19’, ‘The online environment: the new normal’ (ii) ‘Invisibility of male carers’, ‘Carers supporting carers’, ‘The only service you get is lip service: non-existent services’, ‘Knowing your rights’ (iii) ‘Emotional response to the caring role: Feeling devalued’, ‘Emotional response to the caring role: Desperation of caring’, ‘Multiple demands of the caring role.’ Conclusions: Poor or inconsistent access to services and support has been an ongoing difficulty for many family carers. The COVID-19 pandemic has only further intensified these difficulties, increasing family carers' stress. There is an urgent need to design services, such as online support programs, in partnership with family carers that adequately address their needs.

Keywords: intellectual disabilities, family carer, COVID-19, disability

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