Search results for: Loop-Mediated Isothermal Amplification (LAMP)

Authors: Aditya Setyo Rahman, Dwikorita Karnawati, Muzli, Dadang Permana, Sigit Pramono, Fajri Syukur Rahmatullah, Oriza Sativa, Moehajirin, Edy Santoso, Nur Hidayati Oktavia, Ardian Yudhi Octantyo, Robby Wallansha, Juwita Sari Pradita, Nur Fani Habibah, Audia Kaluku, Amelia Chelcea, Yoga Dharma Persada, Anton Sugiharto

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Earthquake vibrations on the surface are not only affected by the magnitude of the earthquake and the distance from the hypocenter but also by the characteristics of the local soil. Variations and changes in soil characteristics from the depth of the bedrock to the surface can cause an amplification of earthquake vibrations that also affect the impact they may have on the surface. Soil characteristics vary widely even at relatively close distances, so for earthquake hazard mapping in cities with earthquake threats, it is necessary to study the characteristics of the local soil on a detailed or micro-scale (microzonation). This study proposes seismic microzonation and liquefaction potential based on microtremor observations. We carried out 143 microtremor observations, and the observation sites were spread across all populated sub-districts in Bengkulu City; the results showed that the dominance of Bengkulu City had medium soil types with a dominant period value of 0.4 < T₀ < 0.6, and there was one location with soft soil characteristics in the river, shaved with T₀ > 0.6. These results correlate with the potential for liquefaction as indicated by a seismic vulnerability index (K𝓰) greater than 5.

Keywords: microtremor, dominant period, microzonation, seismic vulnerability index

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Authors: Mansour F. Hussein, Mohammed A. Alshaikh, Riyadh S. Al-Jumaah, A. GarelNabi, I. Al-Khalifa, Osama B. Mohammed

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Serum samples from 489 male and female camels were tested for antibodies against C. burnetii using indirect enzyme-linked immunosorbent assay (ELISA). Antibodies to C. burnetii were recorded in sera of 252 (51.64%) camels. Significant differences in prevalence were found between male and female camels, juvenile and adult camels, different ecotypes and different sampling locations. 307 camels were simultaneously tested for C. burnetii antibodies by ELISA and indirect immunofluorescence (IFA). Close agreement was found between the results of the two tests. A high prevalence of C. burnetii antibodies was also recorded in milk samples tested by ELISA. Clinical samples from serologically positive camels were subjected to PCR analysis using primers which amplify the repetitive transposon-like and transposase gene regions of C. burnetii. Positive DNA amplification was obtained from both regions, with highest shedding of C. burnetii in faecal samples (27.59%) followed, in descending order, by urine (23.81%), blood (15.85%) and milk (6.5%). The present results indicate that camels are a major reservoir of C. burnetii in Saudi Arabia. The high prevalence of infection in camels, the poor sanitary standards under which the animals are kept and the consumption of raw camel milk indicate that camels could also be a major source of transmission of Q fever to humans in Saudi Arabia.

Keywords: Arabian camel, Camelus dromedarius, Coxiella brunetii, ELISA, immunofluoresence, PCR

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Authors: Rosemary Anibogwu, Kavita Sharma, Karl De Jesus

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Sagebrush is an abundant and naturally occurring plant in the Intermountain West region of the United States. The plant contains an array of bioactive compounds such as flavonoids, terpenoids, sterols, and phenolic acids. It is important to identify and characterize these compounds because Native Americans use sagebrush as herbal medicine. These compounds are also utilized for preventing infection in wounds, treating headaches and colds, and possess antitumor properties. This research is an exploratory study on the sesquiterpene present in the leaves of sagebrush. The leaf foliage was extracted with 100 % chloroform and 100 % methanol. The percentage yield for the crude was considerably higher in chloroform. The Thin Layer Chromatography (TLC) analysis of the crude extracted unveiled a brown band at Rf = 0.25 and a dark brown band at Rf = 0.74, along with three unknown faint bands the 254 nm UV lamp. Furthermore, the two distinct brown (Achillin) and dark brown band (Hydroxyachillin) in TLC were further utilized in the isolation of pure compounds with column chromatography. The structures of Achillin and Hydroxyachillin were elucidated based on extensive spectroscopic analysis, including TLC, High-Performance Liquid Chromatography (HPLC), 1D- and 2D-Nuclear Magnetic Resonance (NMR), and Mass Spectroscopy (MS). The antioxidant activities of crude extract and three pure compounds were evaluated in terms of their peroxyl radical scavenging by Ferric Reducing Ability of Plasma (FRAP) and 1,1-Diphenyl-2-picryl-hydrazyl (DPPH) methods. The crude extract showed the antioxidant activity of 18.99 ± 0.51 µmol TEg -1 FW for FRAP and 11.59 ± 0.38 µmol TEg -1 FW for DPPH. The activities of Achillin, Hydroxyachillin, and Quercetagetin trimethyl ether were 13.03, 15.90 and 14.02 µmol TEg -1 FW respectively for the FRAP assay. The three purified compounds have been submitted to the National Cancer Institute 60 cancer cell line for further study.

Keywords: HPLC, nuclear magnetic resonance spectroscopy, sagebrush, sesquiterpene lactones

Procedia PDF Downloads 131

Authors: Ananthakrishna L. R., Ramesh D., Kumar Wodeyar, Kotresh A. M., Gururaj P. M.

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Malnad Gidda is the indigenous recognized cattle breed of Shivamogga District of Karnataka state, India is known for its disease resistance to many of the infectious diseases. There are 25 LRR (Leucine Rich Repeats) identified in bovine (Bos indicus) TLR9. The amino acid sequence of LRR is deduced to nucleotide sequence in BLASTx bioinformatic online tools. LRR2 to LRR10 are involved in pathogen recognition and binding in human TLR9 which showed a higher degree of nucleotide variations with respect to disease resistance to various pathogens. Hence, primers were designed to amplify the flanking sequences of LRR2 to LRR10, to discover the nucleotide variations if any, in Malnad Gidda breed of Cattle which is associated with disease resistance. The DNA isolated from peripheral blood mononuclear cells of ten Malnad Gidda cattle. A desired and specific amplification product of 0.8 kb was obtained at an annealing temperature of 56.6ᵒC. All the PCR products were sequenced on both sides by gene-specific primers. The sequences were compared with TLR9 sequence of cross breed cattle obtained from NCBI data bank. The sequence analysis between Malnad Gidda and crossbreed cattle revealed no nucleotide variations in the region LRR2 to LRR9 which shows the conserved in pathogen binding domain (LRR) of TLR9.

Keywords: leucine rich repeats, Malnad Gidda, cross breed, TLR9

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Authors: Sukovatov K.Yu., Bezuglova N.N.

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Winter precipitation variability over the Altai Krai was investigated by retrieving temporal patterns. The spectral singular analysis was used to describe the variance distribution and to reduce the precipitation data into a few components (modes). The associated time series were related to large-scale atmospheric and oceanic circulation indices by using lag cross-correlation and wavelet-coherence analysis. GPCC monthly precipitation data for rectangular field limited by 50-550N, 77-880E and monthly climatological circulation index data for the cold season were used to perform SSA decomposition and retrieve statistics for analyzed parameters on the time period 1951-2017. Interannual variability of winter precipitation over the Altai Krai are mostly caused by three natural factors: intensity variations of momentum exchange between mid and polar latitudes over the North Atlantic (explained variance 11.4%); wind speed variations in equatorial stratosphere (quasi-biennial oscillation, explained variance 15.3%); and surface temperature variations for equatorial Pacific sea (ENSO, explained variance 2.8%). It is concluded that under the current climate conditions (Arctic amplification and increasing frequency of meridional processes in mid-latitudes) the second and the third factors are giving more significant contribution into explained variance of interannual variability for cold season atmospheric precipitation over the Altai Krai than the first factor.

Keywords: interannual variability, winter precipitation, Altai Krai, wavelet-coherence

Procedia PDF Downloads 188

Authors: Ergun Sakalar, Kubra Bilgic

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In the last decades, real-time PCR has become a reliable tool preferred to use in many laboratories for pathogen detection. This technique allows for monitoring target amplification via fluorescent molecules besides admit of quantitative analysis by enabling of convert outcomes of thermal cycling to digital data. Sensitivity and traceability of real-time PCR are based on measuring of fluorescence that appears only when fluorescent reporter dye bound to specific target DNA.The fluorescent reporter systems developed for this purpose are divided into two groups. The first group consists of intercalator fluorescence dyes such as SYBR Green, EvaGreen which binds to double-stranded DNA. On the other hand, the second group includes fluorophore-labeled oligonucleotide probes that are separated into three subgroups due to differences in mechanism of action; initial primer-probes such as Cyclicons, Angler®, Amplifluor®, LUX™, Scorpions, and the second one hydrolysis probes like TaqMan, Snake assay, finally hybridization probes, for instance, Molecular Beacons, Hybprobe/FRET, HyBeacon™, MGB-Eclipse, ResonSense®, Yin-Yang, MGB-Pleiades. In addition nucleic acid analogues, an increase of probe affinity to target site is also employed with fluorescence-labeled probes. Consequently, abundant real-time PCR detection chemistries are chosen by researcher according to the field of application, mechanism of action, advantages, and proper structures of primer/probes.

Keywords: fluorescent dye, food safety, molecular probes, nucleic acid analogues

Procedia PDF Downloads 256

Authors: Sana Gul, Sheeba Murad, Aneela Javed

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Introduction: Human Papilloma Virus (HPV) is small DNA virus mostly infecting mucosa and cutaneous keratinocytes. So far, more than 200 Human papillomaviruses are known. HPV have been divided into high- and low-risk on the basis of their oncogenic potential. High risk HPV is considered to be the main etiological cause for cervical cancer. Objective: Current study was designed to screen the local cervical cancer patients from the twin cities of Pakistan for the occurance of high risk HPV. Methodology: A total of 67 formalin fixed paraffin-embedded samples of cervical cancer biopsies were obtained from the government hospitals in Islamabad and Rawalpindi. Cervical cancer biopsies were examined for the presence of HPV DNA. Polymerase chain reaction (PCR) was used for the amplification of a region in the HPV-L1 gene for the general detection of the Papilloma virus and for the genotype specific detection of high risk HPV 16 and 18 using the GP5/GP6 primers and genotype specific primers respectively. Results: HPV DNA was detected in 59 out of 67 samples analyzed. 30 samples showed the presence of HPV16 while 22 samples were positive for HPV 18 . HPV subtype could not be determined in 7 samples. Conclusion: Our results show a strong association between HPV infection and cervical cancer among women in twin cities of Pakistan. One way to minimize the disease burden in relation to HPV infection in Pakistani population is the use of prophylactic vaccines and routine screening. An early diagnosis of HPV infection will allow better health management to reduce the risk of developing cervical cancer.

Keywords: cervical cancer, Pakistan, human papillomavirus, HPV 16

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Authors: Krasimir I. Ivanov, Elitsa N. Kolentsova, Dimitar Y. Dimitrov, Georgi V. Avdeev, Tatyana T. Tabakova

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In recent research copper and manganese systems were found to be the most active in CO and organic compounds oxidation among the base catalysts. The mixed copper manganese oxide has been widely studied in oxidation reactions because of their higher activity at low temperatures in comparison with single oxide catalysts. The results showed that the formation of spinel CuxMn3−xO4 in the oxidized catalyst is responsible for the activity even at room temperature. That is why most of the investigations are focused on the hopcalite catalyst (CuMn2O4) as the best copper-manganese catalyst. Now it’s known that this is true only for CO oxidation, but not for mixture of CO and VOCs. The purpose of this study is to investigate the alumina supported copper-manganese catalysts with different Cu/Mn molar ratio in terms of oxidation of CO, methanol and dimethyl ether. The catalysts were prepared by impregnation of γ-Al2O3 with copper and manganese nitrates and the catalytic activity measurements were carried out in continuous flow equipment with a four-channel isothermal stainless steel reactor. Gas mixtures on the input and output of the reactor were analyzed with a gas chromatograph, equipped with FID and TCD detectors. The texture characteristics were determined by low-temperature (- 196 oС) nitrogen adsorption in a Quantachrome Instruments NOVA 1200e (USA) specific surface area&pore analyzer. Thermal, XRD and TPR analyses were performed. It was established that the active component of the mixed Cu-Mn/γ–alumina catalysts strongly depends on the Cu/Mn molar ratio. Highly active alumina supported Cu-Mn catalysts for CO, methanol and DME oxidation were synthesized. While the hopcalite is the best catalyst for CO oxidation, the best compromise for simultaneous oxidation of all components is the catalyst with Cu/Mn molar ratio 1:5.

Keywords: supported copper-manganese catalysts, CO, VOCs oxidation, combustion of exhaust gases

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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

Procedia PDF Downloads 259

Authors: Morteza Haghi, Cigdem Yilmazbas, Ayse Zeynep Uysal, Melisa Tepedelen, Gozde Turkoz Bakirci

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Today, the increase in plastic waste accumulation is an inescapable consequence of environmental pollution; the disposal of these wastes has caused a significant problem. Variable methods have been utilized; however, biodegradation is the most environmentally friendly and low-cost method. Accordingly, the present study aimed to isolate the bacteria capable of biodegradation of plastics. In doing so, we applied the liquid carbon-free basal medium (LCFBM) prepared with deionized water for the isolation of bacterial species obtained from soil samples taken from the Izmir Menemen region. Isolates forming biofilms on plastic were selected and named (PLB3, PLF1, PLB1B) and subjected to a degradation test. FTIR analysis, 16s rDNA amplification, sequencing, identification of isolates were performed. Finally, at the end of the process, a mass loss of 16.6% in PLB3 isolate and 25% in PLF1 isolate was observed, while no mass loss was detected in PLB1B isolate. Only PLF1 and PLB1B created transparent zones on plastic texture. Considering the FTIR result, PLB3 changed plastic structure by 13.6% and PLF1 by 17%, while PLB1B did not change the plastic texture. According to the 16s rDNA sequence analysis, FLP1, PLB1B, and PLB3 isolates were identified as Streptomyces albogriseolus, Enterobacter cloacae, and Klebsiella pneumoniae, respectively.

Keywords: polyethylene, biodegradation, bacteria, 16s rDNA, FTIR

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Authors: Rabeb Triki, Hassene Djemel, Mounir Baccar

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Surface scraping is a passive heat transfer enhancement technique that is directly used in scraped surface heat exchanger (SSHE). The scraping action prevents the accumulation of the product on the inner wall, which intensifies the heat transfer and avoids the formation of dead zones. SSHEs are widely used in industry for several applications such as crystallization, sterilization, freezing, gelatinization, and many other continuous processes. They are designed to deal with products that are viscous, sticky or that contain particulate matter. This research work presents a three-dimensional numerical simulation of the coupled thermal and hydrodynamic behavior within a SSHE which includes Archimedes’ screw instead of scraper blades. The finite volume Fluent 15.0 was used to solve continuity, momentum and energy equations using multiple reference frame formulation. The process fluid investigated under this study is the pure glycerin. Different geometrical parameters were studied in the case of steady, non-isothermal, laminar flow. In particular, attention is focused on the effect of the conicity of the rotor and the pitch of Archimedes’ screw on temperature and velocity distribution and heat transfer rate. Numerical investigations show that the increase of the number of turns in the screw from five to seven turns leads to amelioration of heat transfer coefficient, and the increase of the conicity of the rotor from 0.1 to 0.15 leads to an increase in the rate of heat transfer. Further studies should investigate the effect of different operating parameters (axial and rotational Reynolds number) on the hydrodynamic and thermal behavior of the SSHE.

Keywords: ANSYS-Fluent, hydrodynamic behavior, scraped surface heat exchange, thermal behavior

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Authors: Eusebio A. Jiménez-Arévalo, Deifilia Ahuatzi-Chacón, Juvencio Galíndez-Mayer, Cleotilde Juárez-Ramírez, Nora Ruiz-Ordaz

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Bendiocarb is a known toxic xenobiotic that presents acute and chronic risks for freshwater invertebrates and estuarine and marine biota; thus, the treatment of water contaminated with the insecticide is of concern. In this paper, a bacterial community with the capacity to grow in bendiocarb as its sole carbon and nitrogen source was isolated by enrichment techniques in batch culture, from samples of a composting plant located in the northeast of Mexico City. Eight cultivable bacteria were isolated from the microbial community, by PCR amplification of 16 rDNA; Pseudoxanthomonas spadix (NC_016147.2, 98%), Ochrobacterium anthropi (NC_009668.1, 97%), Staphylococcus capitis (NZ_CP007601.1, 99%), Bosea thiooxidans. (NZ_LMAR01000067.1, 99%), Pseudomonas denitrificans. (NC_020829.1, 99%), Agromyces sp. (NZ_LMKQ01000001.1, 98%), Bacillus thuringiensis. (NC_022873.1, 97%), Pseudomonas alkylphenolia (NZ_CP009048.1, 98%). NCBI accession numbers and percentage of similarity are indicated in parentheses. These bacteria were regarded as the isolated species for having the best similarity matches. The ability to degrade bendiocarb by the immobilized bacterial community in a packed bed biofilm reactor, using as support volcanic stone fragments (tezontle), was evaluated. The reactor system was operated in batch using mineral salts medium and 30 mg/L of bendiocarb as carbon and nitrogen source. With this system, an overall removal efficiency (η_bend) rounding 90%, was reached.

Keywords: bendiocarb, biodegradation, biofilm reactor, carbamate insecticide

Procedia PDF Downloads 279

Authors: Katalin Gombos, Bence Gálik, Krisztina Ildikó Kalács, Krisztina Gödöny, Ákos Várnagy, József Bódis, Attila Gyenesei, Gábor L. Kovács

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Although non-invasive pre-implantation genetic testing for aneuploidy (NIPGT-A) is potentially appropriate to assess chromosomal ploidy of the embryo, practical application of it in a routine IVF center has not been started in the absence of a recommendation. We developed a comprehensive workflow for a clinically applicable strategy for NIPGT-A based on next-generation sequencing (NGS) technology. We performed MALBAC whole genome amplification and NGS on spent blastocyst culture media of Day 3 embryos fertilized with intra-cytoplasmic sperm injection (ICSI). Spent embryonic culture media of morphologically good quality score embryos were enrolled in further analysis with the blank culture media as background control. Chromosomal abnormalities were identified by an optimized bioinformatics pipeline applying a copy number variation (CNV) detecting algorithm. We demonstrate a comprehensive workflow covering both wet- and dry-lab procedures supporting a clinically applicable strategy for NIPGT-A. It can be carried out within 48 h which is critical for the same-cycle blastocyst transfer, but also suitable for “freeze all” and “elective frozen embryo” strategies. The described integrated approach of non-invasive evaluation of embryonic DNA content of the culture media can potentially supplement existing pre-implantation genetic screening methods.

Keywords: next generation sequencing, in vitro fertilization, embryo assessment, non-invasive pre-implantation genetic testing

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Authors: M. A. Gad, A. Z. Al-Herrawy

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Microsporidia comprises various pathogenic species can infect humans by means of water. Moreover, chlorine disinfection of drinking-water has limitations against this protozoan pathogen. A total of 48 water samples were collected from two drinking water treatment plants having two different filtration systems (slow sand filter and rapid sand filter) during one year period. Samples were collected from inlet and outlet of each plant. Samples were separately filtrated through nitrocellulose membrane (142 mm, 0.45 µm), then eluted and centrifuged. The obtained pellet from each sample was subjected to DNA extraction, then, amplification using genus-specific primer for microsporidia. Each microsporidia-PCR positive sample was performed by two species specific primers for Enterocytozoon bieneusi and Encephalitozoon intestinalis. The results of the present study showed that the percentage of removal for microsporidia through different treatment processes reached its highest rate in the station using slow sand filters (100%), while the removal by rapid sand filter system was 81.8%. Statistically, the two different drinking water treatment plants (slow and rapid) had significant effect for removal of microsporidia. Molecular identification of microsporidia-PCR positive samples using two different primers for Enterocytozoon bieneusi and Encephalitozoon intestinalis showed the presence of the two pervious species in the inlet water of the two stations, while Encephalitozoon intestinalis was detected in the outlet water only. In conclusion, the appearance of virulent microsporidia in treated drinking water may cause potential health threat.

Keywords: removal, efficacy, microsporidia, drinking water treatment plants, PCR

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Authors: Faheema Khan

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In order to investigate the influence of genetic background on salt tolerance in Soybean (Glycine max) ten soybean genotypes released/notified in India were selected. (Pusa-20, Pusa-40, Pusa-37, Pusa-16, Pusa-24, Pusa-22, BRAGG, PK-416, PK-1042, and DS-9712). The 10-day-old seedlings were subjected to 0, 25, 50, 75, 100, 125, and 150 mM NaCl for 15 days. Plant growth, leaf osmotic adjustment, and RAPD analysis were studied. In comparison to control plants, the plant growth in all genotypes was decreased by salt stress, respectively. Salt stress decreased leaf osmotic potential in all genotypes however the maximum reduction was observed in genotype Pusa-24 followed by PK-416 and Pusa-20. The difference in osmotic adjustment between all the genotypes was correlated with the concentrations of ion examined such as Na+ and the leaf proline concentration. These results suggest that the genotypic variation for salt tolerance can be partially accounted for by plant physiological measures. The genetic polymorphisms between soybean genotypes differing in response to salt stress were characterized using 25 RAPD primers. These primers generated a total of 1640 amplification products, among which 1615 were found to be polymorphic. A very high degree of polymorphism (98.30%) was observed. UPGMA cluster analysis of genetic similarity indices grouped all the genotypes into two major clusters. Intra-clustering within the two clusters precisely grouped the 10 genotypes in sub-cluster as expected from their physiological findings. Our results show that RAPD technique is a sensitive, precise and efficient tool for genomic analysis in soybean genotypes.

Keywords: glycine max, NaCl, RAPD, proteomics

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Authors: Mojgan Rabiey, Shyamali Roy, Billy Quilty, Ryan Creeth, George Sundin, Robert W. Jackson

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Bacterial canker is a major disease of Prunus species such as cherry (Prunus avium). It is caused by Pseudomonas syringae species including P. syringae pv. syringae (Pss) and P. syringae pv. morsprunorum race 1 (Psm1) and race 2 (Psm2). Concerns over the environmental impact of, and developing resistance to, copper controls call for alternative approaches to disease management. One method of control could be achieved using naturally occurring bacteriophage (phage) infective to the bacterial pathogens. Phages were isolated from soil, leaf, and bark of cherry trees in five locations in the South East of England. The phages were assessed for their host range against strains of Pss, Psm1, and Psm2. The phages exhibited a differential ability to infect and lyse different Pss and Psm isolates as well as some other P. syringae pathovars. However, the phages were unable to infect beneficial bacteria such as Pseudomonas fluorescens. A subset of 18 of these phages were further characterised genetically (Random Amplification of Polymorphic DNA-PCR fingerprinting and sequencing) and using electron microscopy. The phages are tentatively identified as belonging to the order Caudovirales and the families Myoviridae, Podoviridae, and Siphoviridae, with genetic material being dsDNA. Future research will fully sequence the phage genomes. The efficacy of the phage, both individually and in cocktails, to reduce disease progression in vivo will be investigated to understand the potential for practical use of these phages as biocontrol agents.

Keywords: bacteriophage, pseudomonas, bacterial cancker, biological control

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Authors: Vishnu Raj, Chockalingam Prathap

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The objective of this study was to explore the impact of variation in the exit annular area on the local flow field features and the flame stability of an annular premixed swirl burner (unconfined) operated with premixed n-butane air mixture at equivalence ratio (ϕ) = 1, 1 bar, and 300K. A swirl burner with an axial swirl generator having a swirl number of 1.5 was used. Three different burner heads were chosen to have the exit area increased from 100%, 160%, and 220% resulting in inner and outer diameters and cross-sectional areas as (1) 10mm&15mm, 98mm2 (2) 17.5mm&22.5mm, 157mm2 and (3) 25mm & 30mm, 216mm2. The bulk velocity and Reynolds number based on the hydraulic diameter and unburned gas properties were kept constant at 12 m/s and 4000. (i) Planar PIV with TiO2 seeding particles and (ii) OH* chemiluminescence were used to measure the velocity fields and reaction zones of the swirl flames at 5Hz, respectively. Velocity fields and the jet spreading rates measured at the isothermal and reactive conditions revealed that the presence of a flame significantly altered the flow field in the radial direction due to the gas expansion. Important observations from the flame measurements were: the height and maximum width of the recirculation bubbles normalized by the hydraulic diameter, and the jet spreading angles for the flames for the three exit area cases were: (a) 4.52, 1.95, 28ᵒ, (b) 6.78, 2.37, 34ᵒ, and (c) 8.73, 2.32, 37ᵒ. The lean blowout was also measured, and the respective equivalence ratios were: 0.80, 0.92, and 0.82. LBO was relatively narrow for the 157mm2 case. For this case, particle image velocimetry (PIV) measurements showed that Turbulent Kinetic Energy and turbulent intensity were relatively high compared to the other two cases, resulting in higher stretch rates and narrower lean blowout (LBO).

Keywords: chemiluminescence, jet spreading rate, lean blowout, swirl flow

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Authors: S. Bahrami, A. Rezaie, Z. Boroumand, S. Ghavami

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Neospora caninum is protozoan parasite which can cause a serious disease in dogs and cattle. It has been shown that birds may be a permissive intermediate host for N. caninum since parasite DNA has been detected in tissues from birds. It is showed that embryonated chicken egg can be used as an animal model for experimental infection. The aim of present study was to compare experimental infection of Neospora in chicken and pigeons embryonated eggs. An infection with N. caninum Nc1 isolate was conducted in chicken and pigeons embryonated eggs to evaluate LD50. After calculation of LD50, 2LD50 of tachyzoites were injected to eggs. Macroscopic changes of each embryo were noticed and to investigate the parasite distribution in tissues immunohistochemistry (IHC) and molecular methods were used. In the present study, histopathological changes were considered and sections to those used for histopathological examination including heart, liver, brain and chorioallantoic (CA) membrane were subjected to IHC, too. For PCR procedure, primer pair Np21/Np6 was used for amplification of the Nc5 gene. Pigeon's embryo showed more macroscopic changes than chicken embryo. A hemorrhage of the CA was the main grass lesion. All the infected tissues had histopathological changes. Microscopic examination of tissues revealed acute neosporosis due to hemorrhage, necrosis and infiltration of mononuclear inflammatory cells. Based on IHC and molecular results, the parasite aggregation in the heart was more predominant than in the other tissues. These results reinforce that there is genetic susceptibility to N. caninum in pigeons embryonated eggs like chickens embryonated eggs and provide new insights to research an inexpensive and available animal model for N. caninum.

Keywords: immunohistochemistry, Neospora caninum, PCR, pigeon embryonated egg

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Authors: Ogbonnaya O. Iroanya, Samson T. Fakorede, Osamudiamen J. Edosa, Hadiat A. Azeez

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The human mitochondrial DNA (mtDNA) is about 17 kbp circular DNA fragments found within the mitochondria together with smaller fragments of 1200 bp known as the control region. Knowledge of variation within populations has been employed in forensic and molecular anthropology studies. The study was aimed at investigating the polymorphic nature of the two hypervariable segments (HVS) of the mtDNA, i.e., HVS1 and HVS2, and to determine the haplogroup distribution among individuals resident in Lagos, Southwestern Nigeria. Peripheral blood samples were obtained from sixty individuals who are not related maternally, followed by DNA extraction and amplification of the extracted DNA using primers specific for the regions under investigation. DNA amplicons were sequenced, and sequenced data were aligned and compared to the revised Cambridge Reference Sequence (rCRS) GenBank Accession number: NC_012920.1) using BioEdit software. Results obtained showed 61 and 52 polymorphic nucleotide positions for HVS1 and HVS2, respectively. While a total of three indels mutation were recorded for HVS1, there were seven for HVS2. Also, transition mutations predominate nucleotide change observed in the study. Genetic diversity (GD) values for HVS1 and HVS2 were estimated to be 84.21 and 90.4%, respectively, while random match probability was 0.17% for HVS1 and 0.89% for HVS2. The study also revealed mixed haplogroups specific to the African (L1-L3) and the Eurasians (U and H) lineages. New polymorphic sites obtained from the study are promising for human identification purposes.

Keywords: hypervariable region, indels, mitochondrial DNA, polymorphism, random match probability

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Authors: Xing Wang, Lin Feng, Xuan Zou, Hongchen liu

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Immediate Implantation after tooth extraction is considered to be the ideal way to retain the alveolar bone, but some scholars believe the aesthetic effect in the Early Implantation case are more reliable. In this study, 89 patients were added to this retrospective study up to 5 years. Assessment indicators was including the survival of the implant (peri-implant infection, implant loosening, shedding, crowns and occlusal), aesthetics (color and fullness gums, papilla height, probing depth, X-ray alveolar crest height, the patient's own aesthetic satisfaction, doctors aesthetics score), repair defects around the implant (peri-implant bone changes in height and thickness, whether the use of autologous bone graft, whether to use absorption/repair manual nonabsorbable material), treatment time, cost and the use of antibiotics.The results demonstrated that there is no significant difference in long-term success rate of immediate implantation, early implantation and delayed implantation (p> 0.05). But the results indicated immediate implantation group could get get better aesthetic results after two years (p< 0.05), but may increase the risk of complications and failures (p< 0.05). High-risk indicators include gingival recession, labial bone wall damage, thin gingival biotypes, planting position and occlusal restoration bad and so on. No matter which type of implanting methods was selected, the extraction methods and bone defect amplification techniques are observed as a significant factors on aesthetic effect (p< 0.05).

Keywords: immediate implantation, long-term effects, aesthetics area, dental implants

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Authors: Baljinder Singh, Navneet Sharma

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Chromium is an essential nutrient involved in normal carbohydrate and lipid metabolism. It influences glucose metabolism by potentiating the action as taking part in insulin signal amplification mechanism. A placebo-controlled single blind, prospective study was carried out to investigate the effect of chromium supplementation on blood glucose, HbA1C and lipid profile in newly onset patients with type-2 diabetes. Total 40 newly onset type-2 diabetics were selected and after one month stabilization further randomly divided into two groups viz. study group and placebo group. The study group received 9 gm brewer’s yeast (42 μ Cr) daily and the other placebo group received yeast devoid of chromium for 3 months. Subjects were instructed not to change their normal eating and living habits. Fasting blood glucose, HbA1C and lipid profile were analyzed at beginning and completion of the study. Results revealed that fasting blood glucose level significantly reduced in the subjects consuming yeast supplemented with chromium (197.65±6.68 to 103.68±6.64 mg/dl; p<0.001). HbA1C values improved significantly from 9.51±0.26% to 6.86±0.28%; p<0.001 indicating better glycaemic control. In experimental group total cholesterol, TG and LDL levels were also significantly reduced from 199.66±3.11 to 189.26±3.01 mg/dl; p<0.02, 144.94±8.31 to 126.01±8.26; p<0.05 and 119.19±1.71 to 99.58±1.10; p<0.001 respectively. These data demonstrate beneficial effect of chromium supplementation on glycaemic control and lipid variables in subjects with newly onset type-2 diabetes.

Keywords: type-2 diabetes, chromium, glucose, HbA1C

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Authors: D. S. Malatesh, K. J. Ananda, C. Ansar Kamran, K. Ganesh Udupa

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Dirofilaria repens is a mosquito-borne filarioid nematode of dogs and other carnivores and accidentally affects humans. D. repens is reported in many countries, including India. Subcutaneous dirofilariosis caused by D. repens is a zoonotic disease, widely distributed throughout Europe, Asia, and Africa, with higher prevalence reported in dogs from Sri Lanka (30-60%), Iran (61%) and Italy (21-25%). Dirofilariasis in dogs was diagnosed by detection of microfilariae in blood. Identification of different Dirofilaria species was done by using molecular methods like polymerase chain reaction (PCR). Even though many researchers reported molecular evidence of D. repens across India, to our best knowledge there is no data available on molecular diagnosis of D. repens in dogs and its zoonotic implication in Karnataka state a southern state in India. The aim of the present study was to identify the Dirofilaria species occurring in dogs from Karnataka, India. Out of 310 samples screened for the presence of microfilariae using traditional diagnostic methods, 99 (31.93%) were positive for the presence of microfilariae. Based on the morphometry, the microfilariae were identified as D. repens. For confirmation of species, the samples were subjected to PCR using pan filarial primers (DIDR-F1, DIDR-R1) for amplification of internal transcribed spacer region 2 (ITS2) of the ribosomal DNA. The PCR product of 484 base pairs on agarose gel was indicative of D. repens. Hence, a single PCR reaction using pan filarial primers can be used to differentiate filarial species found in dogs. The present study confirms that dirofilarial species occurring in dogs from Karnataka is D. repens and further sequencing studies are needed for genotypic characterization of D. repens.

Keywords: Dirofilaria repens, molecular characterization, polymerase chain reaction, Karnataka, India

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Authors: Montree Bunruangses, Sonath Bhattacharyya, Somchat Sonasang, Preecha Yupapin

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A two-level system is manipulated by a microstrip add-drop circuit configured as an atom like system for wave-particle behavior investigation when its traveling speed along the circuit perimeter is the speed of light. The entangled pair formed by the upper and lower sideband peaks is bound by the angular displacement, which is given by 0≤θ≤π/2. The control signals associated with 3-peak signal frequencies are applied by the external inputs via the microstrip add-drop multiplexer ports, where they are time functions without the space term involved. When a system satisfies the speed of light conditions, the mass term has been changed to energy based on the relativistic limit described by the Lorentz factor and Einstein equation. The different applied frequencies can be utilized to form the 3-phase torques that can be applied for quantum engines. The experiment will use the two-level system circuit and be conducted in the laboratory. The 3-phase torques will be recorded and investigated for quantum engine driving purpose. The obtained results will be compared to the simulation. The optimum amplification of torque can be obtained by the resonant successive filtering operation. Torque will be vanished when the system is balanced at the stopped position, where |Time|=0, which is required to be a system stability condition. It will be discussed for future applications. A larger device may be tested in the future for realistic use. A synchronous and asynchronous driven motor is also discussed for the warp drive use.

Keywords: quantum engine, relativistic motor, 3-phase torque, atomic engine

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Authors: Adel M. Zakri, Mohammed A. Al-Saleh, Judith. K. Brown, Ali M. Idris

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Betasatellites are small circular single stranded DNA molecules found associated with begomoviruses on field symptomatic plants. Their genome size is about half that of the helper begomovirus, ranging between 1.3 and 1.4 kb. The helper begomoviruses are usually members of the family Geminiviridae. Okra leaves showing vein thickening were collected from okra plants growing in Jazan, Saudi Arabia. Total DNA was extracted from leaves and used as a template to amplify circular DNA using rolling circle amplification (RCA) technology. Products were digested with PstI to linearize the helper viral genome(s), and associated DNA satellite(s), yielding a 2.8kbp and 1.4kbp fragment, respectively. The linearized fragments were cloned into the pGEM-5Zf (+) vector and subjected to DNA sequencing. The 2.8 kb fragment was identified as Cotton leaf curl Gezira virus genome, at 2780bp, an isolate closely related to strains reported previously from Saudi Arabia. A clone obtained from the 1.4 kb fragments he 1.4kb was blasted to GeneBank database found to be a betasatellite. The genome of betasatellite was 1357-bp in size. It was found to be a recombinant containing one fragment (877-bp) that shared 91% nt identity with Cotton leaf curl Gezira betasatellite [KM279620], and a smaller fragment [133--bp) that shared 86% nt identity with Tomato leaf curl Sudan virus [JX483708]. This satellite is thus a recombinant between a malvaceous-infecting satellite and a solanaceous-infecting begomovirus.

Keywords: begomovirus, betasatellites, cotton leaf curl Gezira virus, okra plants

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Authors: Ha T. T. Ly, Truc B. Truc, Hai N. Truong, Mai P. T. Nguyen, Ngoc D. Ngo, Khanh V. Tran, Hai T. Le

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Beta thalassemia is one of the most common inherited blood disorders, which is characterized by decreased or absent in beta globin expression. Patients with Beta thalassemia whose anemia is not so severe as to necessitate transfusions are said to have thalassemia intermedia. Objective: The goal of this study is prenatal diagnosis for pregnancy woman with Beta thalassemia intermedia and her husband with Beta thalassemia carrier at high risk of Beta thalassemia major in Northern of Vietnam. Material and method: The family has a 6 years-old compound heterozygous thalassemia major for CD71/72(+A) and Hbb:c. -78A>G/nt-28(A>G) male child. The father was heterozygous for CD71/72(+A) mutation which is Beta plus type and the mother was compound heterozygosity of two different variants, namely, Hbb: c. -78A>G/nt-28(A>G) and CD26(A-G) HbE. Prenatal Beta thalassemia mutation detection in fetal DNA was carried out using multiplex Amplification-refractory mutation system ARMS-PCR and confirmed by direct Sanger-sequencing Hbb gene. Prenatal diagnoses were perfomed by amniotic fluid sampling from pregnant woman in the 16-18th week of pregnancy after the genotypes of parents of the probands were identified. Result: When amniotic fluid sample was analyzed for Beta globin gene (Hbb), we found that the genotype is heterozygous for CD71/72(+A) and CD26(A-G) HbE. This genotype is different from the 1st child of this family. Conclusion: Prenatal diagnosis helps the parents to know the genotype and the thalassemia status of the fetus, so they can have early decision on their pregnancy. Genetic diagnosis provided a useful method in diagnosis for familial members in pedigree, genetic counseling and prenatal diagnosis.

Keywords: beta thalassemia intermedia, Hbb gene, pedigree, prenatal diagnosis

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Authors: Muhammad S. Sajid, A. Iqbal, A. Kausar, M. Jawad-ul-Hassan, Z. Iqbal, Hafiz M. Rizwan, M. Saqib

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Among the ixodid ticks, species of genus Hyalomma are of prime importance as they can survive in harsh conditions better than those of other species. Similarly, among various tick-borne pathogens, Theileria (T.) annulata, the causative agent of tropical theileriosis in large ruminants, is responsible for reduced productivity and ultimately substantial economic losses due to morbidity and mortality. The present study was planned to screening of vector ticks through molecular techniques for determination of tick-borne theileriosis in district Toba Tek Singh (T. T. Singh), Punjab, Pakistan. For this purpose, among the collected ticks (n = 2252) from livestock and their microclimate, Hyalomma spp. were subjected to dissection for procurement of salivary glands (SGs) and formation of pool (averaged 8 acini in each pool). Each pool of acini was used for DNA extraction, quantification and primer-specific amplification of 18S rRNA of Theileria (T.) annulata. The amplicons were electrophoresed using 1.8% agarose gel following by imaging to identify the band specific for T. annulata. For confirmation, the positive amplicons were subjected to sequencing, BLAST analysis and homology search using NCBI software. The number of Theileria-infected acini was significantly higher (P < 0.05) in female ticks vs male ticks, infesting ticks vs questing ticks and riverine-collected vs non-riverine collected. The data provides first attempt to quantify the vectoral capacity of ixodid ticks in Pakistan for T. annulata which can be helpful in estimation of risk analysis of theileriosis to the domestic livestock population of the country.

Keywords: Hyalomma anatolicum, ixodids, PCR, Theileria annulata

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Authors: Meenu Kaushik, Ayon K. Bandhoyadhayay, Lalit M. Joshi

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Inductive output tubes (IOTs) are widely used as microwave power amplifiers for broadcast and scientific applications. It is capable of amplifying radio frequency (RF) power with very good efficiency. Its compactness, reliability, high efficiency, high linearity and low operating cost make this device suitable for various applications. The device consists of an integrated structure of electron gun and RF cavity, collector and focusing structure. The working principle of IOT is a combination of triode and klystron. The cathode lies in the electron gun produces a stream of electrons. A control grid is placed in close proximity to the cathode. Basically, the input part of IOT is the integrated structure of gridded electron gun which acts as an input cavity thereby providing the interaction gap where the input RF signal is applied to make it interact with the produced electron beam for supporting the amplification phenomena. The paper presents the design, fabrication and testing of a radial re-entrant cavity for implementing in the input structure of IOT at 350 MHz operating frequency. The model’s suitability has been discussed and a generalized mathematical relation has been introduced for getting the proper transverse magnetic (TM) resonating mode in the radial narrow gap RF cavities. The structural modeling has been carried out in CST and SUPERFISH codes. The cavity is fabricated with the Aluminum material and the RF characterization is done using vector network analyzer (VNA) and the results are presented for the resonant frequency peaks obtained in VNA.

Keywords: inductive output tubes, IOT, radial cavity, coaxial cavity, particle accelerators

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Authors: T. Srinivasa Krishna, K. Narendra, K. Thomas, S. S. Raju, B. Munibhadrayya

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The densities, speeds of sound and refractive index of the binary mixture of ionic liquid (IL) 1-Butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([BMIM][Imide]) and Pyrrolidin-2-one(PY) was measured at atmospheric pressure, and over the range of temperatures T= (298.15 -323.15)K. The excess molar volume, excess isentropic compressibility, excess speed of sound, partial molar volumes, and isentropic partial molar compressibility were calculated from the values of the experimental density and speed of sound. From the experimental data excess thermal expansion coefficients and isothermal pressure coefficient of excess molar enthalpy at 298.15K were calculated. The results were analyzed and were discussed from the point of view of structural changes. Excess properties were calculated and correlated by the Redlich–Kister and the Legendre polynomial equation and binary coefficients were obtained. Values of excess partial volumes at infinite dilution for the binary system at different temperatures were calculated from the adjustable parameters obtained from Legendre polynomial and Redlich–Kister smoothing equation. Deviation in refractive indices ΔnD and deviation in molar refraction, ΔRm were calculated from the measured refractive index values. Equations of state and several mixing rules were used to predict refractive indices of the binary mixtures and compared with the experimental values by means of the standard deviation and found to be in excellent agreement. By using Prigogine–Flory–Patterson (PFP) theory, the above thermodynamic mixing functions have been calculated and the results obtained from this theory were compared with experimental results.

Keywords: density, refractive index, speeds of sound, Prigogine-Flory-Patterson theory

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Authors: Mun Yee Chan, Sin Ming Goh, Lisa Gaik Ai Ong

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Approximately 10,000 different types of dyes and pigments are being used in various industrial applications yearly, which include the textile and printing industries. However, these dyes are difficult to degrade naturally once they enter the aquatic system. Their high persistency in natural environment poses a potential health hazard to all form of life. Hence, there is a need for alternative dye removal strategy in the environment via bioremediation. In this study, fungi laccase is investigated via commercial agar dyes plates and submerged fermentation to explore the application of fungi laccase in textile dye wastewater treatment. Two locally isolated basidiomycetes were screened for laccase activity using media added with commercial dyes such as 2, 2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS), guaiacol and Remazol Brillant Blue R (RBBR). Isolate TBB3 (1.70±0.06) and EL2 (1.78±0.08) gave the highest results for ABTS plates with the appearance of greenish halo on around the isolates. Submerged fermentation performed on Isolate TBB3 with the productivity 3.9067 U/ml/day, whereas the laccase activity for Isolate EL2 was much lower (0.2097 U/ml/day). As isolate TBB3 showed higher laccase production, it was subjected to molecular characterization by DNA isolation, PCR amplification and sequencing of ITS region of nuclear ribosomal DNA. After being compared with other sequences in National Center for Biotechnology Information (NCBI database), isolate TBB3 is probably from species Trametes hirsutei. Further research work can be performed on this isolate by upscale the production of laccase in order to meet the demands of the requirement for higher enzyme titer for the bioremediation of textile dyes.

Keywords: bioremediation, dyes, fermentation, laccase

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Authors: S. Hannah Elizabeth, A. Panneerselvam

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Objective: The main objective was to evaluate the degradative properties of Bacillus cereus sp TUHP2 isolated from TNT-Polluted soils in the Vellore District, Tamil Nadu, India. Methods: Among the 3 bacterial genera isolated from different soil samples, one potent TNT degrading strain Bacillus cereus sp TUHP2 was identified. The morphological, physiological and the biochemical properties of the strain Bacillus cereus sp TUHP2 was confirmed by conventional methods and genotypic characterization was carried out using 16S r-DNA partial gene amplification and sequencing. The broken down by products of DNT in the extract was determined by Gas Chromatogram- Mass spectrometry (GC-MS). Supernatant samples from the broth studied at 24 h interval were analyzed by HPLC analysis and the effect on various nutritional and environmental factors were analysed and optimized for the isolate. Results: Out of three isolates one strain TUHP2 were found to have potent efficiency to degrade TNT and revealed the genus Bacillus. 16S rDNA gene sequence analysis showed highest homology (98%) with Bacillus cereus and was assigned as Bacillus cereus sp TUHP2. Based on the energy of the predicted models, the secondary structure predicted by MFE showed the more stable structure with a minimum energy. Products of TNT Transformation showed colour change in the medium during cultivation. TNT derivates such as 2HADNT and 4HADNT were detected by HPLC chromatogram and 2ADNT, 4ADNT by GC/MS analysis. Conclusion: Hence this study presents the clear evidence for the biodegradation process of TNT by strain Bacillus cereus sp TUHP2.

Keywords: bioremediation, biodegradation, biotransformation, sequencing

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