Scholarly

Sustainable Control of Taro Beetles via Scoliid Wasps and Metarhizium anisopliae

Year: 2014 Volume: 8 Issue: 6 642 - 647 Pages

Abstract: Taro Scarab beetles (Papuana uninodis, Coleoptera: Scarabaeidae) inflict severe damage on important root crops and plants such as Taro or Cocoyam, yam, sweet potatoes, oil palm and coffee tea plants across Africa and Asia resulting in economic hardship and starvation in some nations. Scoliid wasps and Metarhizium anisopliae fungus - bio-control agents; are shown to be able to control the population of Scarab beetle adults and larvae using a newly created simulation model based on non-linear ordinary differential equations that track the populations of the beetle life cycle stages: egg, larva, pupa, adult and the population of the scoliid parasitoid wasps, which attack beetle larvae. In spite of the challenge driven by the longevity of the scarab beetles, the combined effect of the larval wasps and the fungal bio-control agent is able to control and drive down the population of both the adult and the beetle eggs below the environmental carrying capacity within an interval of 120 days, offering the long term prospect of a stable and eco-friendly environment; where the population of scarab beetles is: regulated by parasitoid wasps and beneficial soil saprophytes.

Enzyme Involvement in the Biosynthesis of Selenium Nanoparticles by Geobacillus wiegelii Strain GWE1 Isolated from a Drying Oven

Year: 2014 Volume: 8 Issue: 6 637 - 641 Pages

Abstract: The biosynthesis of nanoparticles by microorganisms, on the contrary to chemical synthesis, is an environmentally-friendly process which has low energy requirements. In this investigation, we used the microorganism Geobacillus wiegelii, strain GWE1, an aerobic thermophile belonging to genus Geobacillus, isolated from a drying oven. This microorganism has the ability to reduce selenite evidenced by the change of color from colorless to red in the culture. Elemental analysis and composition of the particles were verified using transmission electron microscopy and energy-dispersive X-ray analysis. The nanoparticles have a defined spherical shape and a selenium elemental state. Previous experiments showed that the presence of the whole microorganism for the reduction of selenite was not necessary. The results strongly suggested that an intracellular NADPH/NADH-dependent reductase mediates selenium nanoparticles synthesis under aerobic conditions. The enzyme was purified and identified by mass spectroscopy MALDI-TOF TOF technique. The enzyme is a 1-pyrroline-5-carboxylate dehydrogenase. Histograms of nanoparticles sizes were obtained. Size distribution ranged from 40-160 nm, where 70% of nanoparticles have less than 100 nm in size. Spectroscopic analysis showed that the nanoparticles are composed of elemental selenium. To analyse the effect of pH in size and morphology of nanoparticles, the synthesis of them was carried out at different pHs (4.0, 5.0, 6.0, 7.0, 8.0). For thermostability studies samples were incubated at different temperatures (60, 80 and 100 ºC) for 1 h and 3 h. The size of all nanoparticles was less than 100 nm at pH 4.0; over 50% of nanoparticles have less than 100 nm at pH 5.0; at pH 6.0 and 8.0 over 90% of nanoparticles have less than 100 nm in size. At neutral pH (7.0) nanoparticles reach a size around 120 nm and only 20% of them were less than 100 nm. When looking at temperature effect, nanoparticles did not show a significant difference in size when they were incubated between 0 and 3 h at 60 ºC. Meanwhile at 80 °C the nanoparticles suspension lost its homogeneity. A change in size was observed from 0 h of incubation at 80ºC, observing a size range between 40-160 nm, with 20% of them over 100 nm. Meanwhile after 3 h of incubation at size range changed to 60-180 nm with 50% of them over 100 nm. At 100 °C the nanoparticles aggregate forming nanorod structures. In conclusion, these results indicate that is possible to modulate size and shape of biologically synthesized nanoparticles by modulating pH and temperature.

The Effect of Drought Stress on Grain Yield, Yield Components and Protein Content of Durum Wheat Cultivars in Ilam Province, Iran

Year: 2014 Volume: 8 Issue: 6 631 - 636 Pages

Abstract: In order to study the effect of drought stress on grain yield, yield components and associated traits of durum wheat cultivars, an experiment was done as split plot arrangement using randomized complete block design with three replications in Ilam province, Iran in 2009-2010 cropping season. Different levels of irrigation (Full irrigation, drought stress at stem elongation, Flowering and grain formation stages) were considered as a main plot and three durum wheat cultivars (Yavaros, Seimareh and Karkheh) were assigned as a sub plot. The results showed that drought stress was significant on grain yield, spike.m-2, grain. Spike-1, 1000-grain weight, biological yield, harvest index and protein content. Drought stress at all stages caused a loss in grain yield and its components. Full irrigation had the highest grain yield and yield components. Drought stress at stem elongation, flowering and grain formation stages caused a reduction in spike.m-2, grain.spike-1 and 1000-grain weight, respectively. Protein content was significantly affected by drought stress. The highest protein content was obtained from drought stress at grain formation stage. Cultivars had an influence on grain yield and yield components. Yavaros and Seimareh cultivars had the highest and lowest grain yield, respectively. Interaction effect between drought stress and cultivar had a significant effect on grain and yield components. Full irrigation and Yavaros cultivar had the highest grain yield and drought stress at grain formation stage and Seimareh cultivar had the lowest grain yield, respectively.

Influence of Culture Conditions on the Growth and Fatty Acid Composition of Green Microalgae Oocystis rhomboideus, Scenedesmus obliquus, Dictyochlorella globosa

Year: 2014 Volume: 8 Issue: 6 626 - 630 Pages

Abstract: Microalgae due to the ability to accumulate high levels of practically valuable polyunsaturated fatty acids attract attention as a promising raw material for commercial products. The features of the growth processes of cells green protococcal microalgae Oocystis rhomboideus, Scenedesmus obliquus, Dictyochlorella globosa at cultivation in different nutritional mediums were determined. For the rapid accumulation of biomass, combined with high productivity of total lipids fraction yield recommended to use the Fitzgerald medium (Scenodesmus obliquus, Oocystis rhomboideus) and/or Bold medium (Dictyochlorella globosa). Productivity of lipids decreased in sequence Dictyochlorella globosa > Scenodesmus obliquus > Oocystis rhomboideus. The bulk of fatty acids fraction of the total lipids is unsaturated fatty acids, which accounts for 70 to 83% of the total number of fatty acids. The share of monoenic acids accounts from 18 to 34%, while the share of unsaturated fatty acids - from 44 to 62% of the total number of unsaturated fatty acids fraction. Among the unsaturated acids dominate α-linolenic acid (C18:3n-3), hexadecatetraenic acid (C16:4) and linoleic acid (C18:2).

Determination of Vitamin C (Ascorbic Acid) in Orange Juices Product

Year: 2014 Volume: 8 Issue: 6 623 - 625 Pages

Authors:
Wanida Wonsawat

Abstract: This research describes a voltammetric approach to determine amounts of vitamin C (Ascorbic acid) in orange juice sample, using three screen printed electrode. The anodic currents of vitamin C were proportional to vitamin C concentration in the range of 0 – 10.0 mM with the limit of detection of 1.36 mM. The method was successfully employed with 2 µL of the working solution dropped on the electrode surface. The proposed method was applied for the analysis of vitamin C in packed orange juice without sample purification or complexion of sample preparation step.

Inhibitory Effects of Ambrosia trifida L. on the Development of Root Hairs and Protein Patterns of Radicles

Year: 2014 Volume: 8 Issue: 6 619 - 622 Pages

Abstract: Ambrosia trifida L. is designated as invasive alien species by the Act on the Conservation and Use of Biodiversity by the Ministry of Environment, Korea. The purpose of present paper was to investigate the inhibitory effects of aqueous extracts of A.trifida on the development of root hairs of Triticum aestivum L., and Allium tuberosum Rottler ex Spreng and the electrophoretic protein patterns of their radicles. The development of root hairs was inhibited by increasing of aqueous extract concentrations. Through SDS-PAGE, the electrophoretic protein bands of extracted proteins from their radicles were appeared in controls, but protein bands of specific molecular weight disappeared or weakened in treatments. In conclusion, inhibitory effects of A. trifida made two receptor species changed morphologically, and at the molecular level in early growth stage.

The Effect of Sea Buckthorn (Hippophae rhamnoides L.) Berries on Some Quality Characteristics of Cooked Pork Sausages

Year: 2014 Volume: 8 Issue: 6 615 - 618 Pages

Abstract: The aim of this study was to analyze selected quality characteristics of cooked pork sausages manufactured with the addition of Sea buckthorn (Hippophae rhamnoides L.) berries preparations. Stuffings of model sausages consisted of pork, backfat, water and additives such a curing salt and sodium isoascorbate. Functional additives used in production process were two preparations obtained from dried Sea buckthorn berries in form of powder and brew. Powder of dried berries was added in amount of 1 and 3 g, while water infusion as a replacement of 50 and 100% ice water included in meat products formula. Control samples were produced without functional additives. Experimental stuffings were heat treated in water bath and stored for 4 weeks under cooled conditions (4±1ºC). Physical parameters of colour, texture profile and technological parameters as acidity, weight losses and water activity were estimated. The effect of Sea buckthorn berries preparations on lipid oxidation during storage of final products was determine by TBARS method. Studies have shown that addition of Sea buckthorn preparations to meat-fatty batters significant (P≤0.05) reduced the pH values of sausages samples after thermal treatment. Moreover, the addition of berries powder caused significant differences (P ≤ 0.05) in weight losses after cooking process. Analysis of results of texture profile analysis indicated, that utilization of infusion prepared from Sea buckthorn dried berries caused increase of springiness, gumminess and chewiness of final meat products. At the same time, the highest amount of Sea buckthorn berries powder in recipe caused the decrease of all measured texture parameters. Utilization of experimental preparations significantly decreased (P≤0.05) lightness (L* parameter of color) of meat products. Simultaneously, introduction of 1 and 3 grams of Sea buckthorn berries powder to meat-fatty batter increased redness (a* parameter) of samples under investigation. Higher content of substances reacting with thiobarbituric acid was observed in meat products produced without functional additives. It was observed that powder of Sea buckthorn berries added to meat-fatty batters caused higher protection against lipid oxidation in cooked sausages.

Prevalence and Fungicidal Activity of Endophytic Micromycetes of Plants in Kazakhstan

Year: 2014 Volume: 8 Issue: 6 609 - 614 Pages

Abstract: Endophytic microorganisms are presented in plants of different families growing in the foothills and piedmont plains of Trans-Ili Alatau. It was found that the maximum number of endophytic micromycetes is typical to the Fabaceae family. The number of microscopic fungi in the roots reached (145.9±5.9)×103 CFU/g of plant tissue; yeasts - (79.8±3.5)×102 CFU/g of plant tissue. Basically, endophytic microscopic fungi are typical for underground parts of plants. In contrast, yeasts more infected aboveground parts of plants. Small amount of micromycetes is typical to inflorescence and fruits. Antagonistic activity of selected micromycetes against Fusarium graminearum, Cladosporium sp., Phytophtora infestans and Botrytis cinerea phytopathogens was detected. Strains with a broad, narrow and limited range of action were identified. For further investigations Rh2 and T7 strains were selected, they are characterized by a broad spectrum of fungicidal activity and they formed the large inhibition zones against phytopathogens. Active antagonists are attributed to the Rhodotorula mucilaginosa and Beauveria bassiana species.

Influence of Culturing Conditions on Biomass Yield, Total Lipid, and Fatty Acid Composition of Some Filamentous Fungi

Year: 2014 Volume: 8 Issue: 6 605 - 608 Pages

Abstract: In this work the effect of culturing conditions of filamentous fungi Penicillium raistrickii, Penicillium anatolicum, Fusarium sp. on biomass yield, the content of total lipids and fatty acids was studied. It has been established that in time the process of lipids accumulation correlated with biomass growth of cultures, reaching maximum values in stationary growth phase. Biomass yield and accumulation of general lipids was increased by adding zinc to the culture medium. The more intensive accumulation of biomass and general lipids was observed at temperature 18°C. Lowering the temperature of culturing has changed the ratio of saturated: Unsaturated fatty acids in the direction of increasing the latter.

Evaluation of Antioxidant Activities of Cabbage (Brassica oleracea L. var. capitata L.)

Year: 2014 Volume: 8 Issue: 6 602 - 604 Pages

Authors:
Rutanachai Thaipratum

Abstract: At present, it is widely-known that free radicals are the causes of illness such as cancers, coronary heart disease, Alzheimer’s disease and aging. One method of protection from free radical is the consumption of antioxidant-containing foods or herbs. Several analytical methods have been used for qualitative and quantitative determination of antioxidants. This project aimed to evaluate antioxidant activity of ethanolic and aqueous extracts from cabbage (Brassicca oleracea L. var. capitata L.) measured by DPPH and Hydroxyl radical scavenging method. The results show that averaged antioxidant activity measured in ethanolic extract (µmol Ascorbic acid equivalent/g fresh mass) were 7.316 ± 0.715 and 4.66 ± 1.029 as determined by DPPH and Hydroxyl radical scavenging activity assays respectively. Averaged antioxidant activity measured in aqueous extract (µmol Ascorbic acid equivalent/g fresh mass) were 15.141 ± 2.092 and 4.955 ± 1.975 as determined by DPPH and Hydroxyl radical scavenging activity assays respectively.

Systematic Identification and Quantification of Substrate Specificity Determinants in Human Protein Kinases

Year: 2014 Volume: 8 Issue: 6 592 - 601 Pages

Abstract: Protein kinases participate in a myriad of cellular processes of major biomedical interest. The in vivo substrate specificity of these enzymes is a process determined by several factors, and despite several years of research on the topic, is still far from being totally understood. In the present work, we have quantified the contributions to the kinase substrate specificity of i) the phosphorylation sites and their surrounding residues in the sequence and of ii) the association of kinases to adaptor or scaffold proteins. We have used position-specific scoring matrices (PSSMs), to represent the stretches of sequences phosphorylated by 93 families of kinases. We have found negative correlations between the number of sequences from which a PSSM is generated and the statistical significance and the performance of that PSSM. Using a subset of 22 statistically significant PSSMs, we have identified specificity determinant residues (SDRs) for 86% of the corresponding kinase families. Our results suggest that different SDRs can function as positive or negative elements of substrate recognition by the different families of kinases. Additionally, we have found that human proteins with known function as adaptors or scaffolds (kAS) tend to interact with a significantly large fraction of the substrates of the kinases to which they associate. Based on this characteristic we have identified a set of 279 potential adaptors/scaffolds (pAS) for human kinases, which is enriched in Pfam domains and functional terms tightly related to the proposed function. Moreover, our results show that for 74.6% of the kinase–pAS association found, the pAS colocalize with the substrates of the kinases they are associated to. Finally, we have found evidence suggesting that the association of kinases to adaptors and scaffolds, may contribute significantly to diminish the in vivo substrate crossed-specificity of protein kinases. In general, our results indicate the relevance of several SDRs for both the positive and negative selection of phosphorylation sites by kinase families and also suggest that the association of kinases to pAS proteins may be an important factor for the localization of the enzymes with their set of substrates.

Immobilization of Lipase Enzyme by Low Cost Material: A Statistical Approach

Year: 2014 Volume: 8 Issue: 6 588 - 591 Pages

Abstract: Immobilization of lipase enzyme produced from palm oil mill effluent (POME) by the activated carbon (AC) among the low cost support materials was optimized. The results indicated that immobilization of 94% was achieved by AC as the most suitable support material. A sequential optimization strategy based on a statistical experimental design, including one-factor-at-a-time (OFAT) method was used to determine the equilibrium time. Three components influencing lipase immobilization were optimized by the response surface methodology (RSM) based on the face-centered central composite design (FCCCD). On the statistical analysis of the results, the optimum enzyme concentration loading, agitation rate and carbon active dosage were found to be 30 U/ml, 300 rpm and 8 g/L respectively, with a maximum immobilization activity of 3732.9 U/g-AC after 2 hrs of immobilization. Analysis of variance (ANOVA) showed a high regression coefficient (R2) of 0.999, which indicated a satisfactory fit of the model with the experimental data. The parameters were statistically significant at p

Assessment of Susceptibility of the Poultry Red Mite, Dermanyssus gallinae (Acari: Dermanyssidae) to Some Plant Preparations with Focus on Exposure Time

Year: 2014 Volume: 8 Issue: 6 584 - 587 Pages

Abstract: Plant preparations from thyme and garlic have been shown to be effective acaricides against the poultry red mite, Dermanyssus gallinae. In a layer house with a history of D. gallinae problem, mites were detected in the monitoring traps for the first time and number of them was counted. Then, some rows of layer house was sprayed twice using a concentration of 0.21 mg/cm2 thyme essential oil and 0.07 mg/cm2 garlic juice and a similar row was used as an untreated control group. Red mite traps made of cardboard were used to assess the mite density during days 1 and 7 after treatment and always removed after 24 h. the collected mites were counted and the efficacy against all mite stages (larvae, nymphs and adults) was calculated. Results showed that on day 1 and 7 after the administration of garlic extract efficacy rate was 92.05% and 74.62%, respectively. Moreover, efficacy rate on day 1 and 7 was 89.4% and 95.37% when treatment was done with thyme essential oil. It is concluded that using garlic juice to control of D. gallinae is more effective on short time. But thyme essential oil has a long time effect in compare to garlic preparation.

Statistical Modeling for Permeabilization of a Novel Yeast Isolate for β-Galactosidase Activity Using Organic Solvents

Year: 2014 Volume: 8 Issue: 6 578 - 583 Pages

Abstract: The hydrolysis of lactose using β-galactosidase is one of the most promising biotechnological applications, which has wide range of potential applications in food processing industries. However, due to intracellular location of the yeast enzyme, and expensive extraction methods, the industrial applications of enzymatic hydrolysis processes are being hampered. The use of permeabilization technique can help to overcome the problems associated with enzyme extraction and purification of yeast cells and to develop the economically viable process for the utilization of whole cell biocatalysts in food industries. In the present investigation, standardization of permeabilization process of novel yeast isolate was carried out using a statistical model approach known as Response Surface Methodology (RSM) to achieve maximal b-galactosidase activity. The optimum operating conditions for permeabilization process for optimal β-galactosidase activity obtained by RSM were 1:1 ratio of toluene (25%, v/v) and ethanol (50%, v/v), 25.0 oC temperature and treatment time of 12 min, which displayed enzyme activity of 1.71 IU /mg DW.

Utilization of Agro-Industrial Byproducts for Bacteriocin Production Using Newly Isolated Enterococcus faecium BS13

Year: 2014 Volume: 8 Issue: 6 573 - 577 Pages

Abstract: Microbial production of antimicrobials as biopreservatives is the major area of focus nowadays due to increased interest of consumers towards natural and safe preservation of ready to eat food products. The agro-industrial byproduct based medium and optimized process conditions can contribute in economical production of bacteriocins. Keeping this in view, the present investigation was carried out on agro-industrial byproducts utilization for the production of bacteriocin using Enterococcus faecium BS13 isolated from local fermented food. Different agro-industrial byproduct based carbon sources (whey, potato starch liquor, kinnow peel, deoiledrice bran and molasses), nitrogen sources (soya okra, pea pod and corn steep liquor), metal ions and surfactants were tested for optimal bacteriocin production. The effect of various process parameters such as pH, temperature, inoculum level, agitation and time were also tested on bacteriocin production. The optimized medium containing whey, supplemented with 4%corn steep liquor and polysorbate-80 displayed maximum bacteriocin activity with 2% inoculum, at pH 6.5, temperature 40oC under shaking conditions (100 rpm).

Bacteria Flora in the Gut and Respiratory Organs of Clarias gariepinus in Fresh and Brackish Water Habitats of Ondo State, South/West Nigeria

Year: 2014 Volume: 8 Issue: 6 569 - 572 Pages

Abstract: Bacteria flora of Clarias gariepinus collected from two natural habitats namely Owena River (freshwater) and Igbokoda lagoon (brackish water) were examined using standard microbiological procedures. Thirteen bacterial species were identified. The result indicated that from the identified bacteria isolated, Vibrio sp, Proteus sp. Shigella sp. and E. coli were present in both habitats (fresh and brackish waters). Others were habitat-selective such as Salmonella sp., Pseudomonas sp, Enterococcus sp, Staphylococcus sp. that were found only in freshwater habitat. While Branhamella sp, Streptococcus sp. and Micrococcus sp. were found in brackish water habitat. Bacteria load from Owena river (freshwater) was found to be the highest load recorded at 6.21 x 104cfu. T-test analysis also revealed that there was a marked significant difference between bacterial load in guts of sampled Clarias from fresh water and brackish water habitats.

Toxicity of Bisphenol-A: Effects on Health and Regulations

Year: 2014 Volume: 8 Issue: 6 564 - 568 Pages

Abstract: Bisphenol-A (BPA) is one of the highest volume chemicals produced worldwide in the plastic industry. This compound is mostly used in producing polycarbonate plastics that are often used for food and beverage storage, and BPA is also a component of epoxy resins that are used to line food and beverage containers. Studies performed in this area indicated that BPA could be extracted from such products while they are in contact with food. Therefore, BPA exposure is presumed. In this paper, the chemical structure of BPA, factors affecting BPA migration to food and beverages, effects on health, and recent regulations will be reviewed.

Laboratory Evaluation of Bacillus subtilis Bioactivity on Musca domestica (Linn) (Diptera: Muscidae) Larvae from Poultry Farms in South Western Nigeria

Year: 2014 Volume: 8 Issue: 6 561 - 563 Pages

Authors:
Funmilola O. Omoya

Abstract: Muscid flies are known to be vectors of disease agents and species that annoy humans and domesticated animals. An example of these flies is Musca domestica (house fly) whose adult and immature stages occur in a variety of filthy organic substances including household garbage and animal manures. They contribute to microbial contamination of foods. It is therefore imperative to control these flies as a result of their role in Public health. The second and third instars of Musca domestica (Linn) were infected with varying cell loads of Bacillus subtilis in vitro for a period of 48 hours to evaluate its larvicidal activities. Mortality of the larvae increased with incubation period after treatment with the varying cell loads. Investigation revealed that the second instars larvae were more susceptible to treatment than the third instars treatments. Values obtained from the third instar group were significantly different (P

SEM Analysis of the Effectiveness of the Acid Etching on Cat Enamel

Year: 2014 Volume: 8 Issue: 6 558 - 560 Pages

Abstract: The aim of this paper is to summarize the literature on micromorphology and composition of the enamel of the cat and present an original experiment by SEM on how it responds to the etching with ortophosphoric acid for the time recommended in the veterinary literature (30", 45", 60"), derived from research and experience on human enamel; 21 teeth of cat were randomly divided into three groups of 7 (A, B, C): Group A was subjected to etching for 30 seconds by means of orthophosphoric acid to 40% on a circular area with diameter of about 2mm of the enamel coronal; the Groups B and C had the same treatment but, respectively, for 45 and 60 seconds. The samples obtained were observed by SEM to constant magnification of 1000x framing, in particular, the border area between enamel exposed and not exposed to etching to highlight differences. The images were subjected to the analysis of three blinded experienced operators in electron microscopy. In the enamel of the cat the etching for the times considered is not optimally effective for the purpose adhesives and the presence of a thick prismless layer could explain this situation. To improve this condition may clinically in the likeness of what is proposed for the enamel of human deciduous teeth: a bevel or a chamfer of 1 mm on the contour of the cavity to discover the prismatic enamel and increase the bonding surface.

Keywords:
SEM
Cat
Enamel

Periodontal Disease or Cement Disease? New Frontier in the Treatment of Periodontal Disease in Dogs

Year: 2014 Volume: 8 Issue: 6 552 - 557 Pages

Abstract: A group of 10 dogs (group A) with Periodontal Disease in the third stage, were subjected to regenerative therapy of periodontal tissues, by use of nano hydroxy apatite (NHA). These animals induced by general anesthesia, where treated by ultrasonic scaling, root planning, and at the end by a mucogingival flap in which it was applied NHA. The flap was closed and sutured with simple steps. Another group of 10 dogs (group B), control group, was treated only by scaling and root planning. No patient was subjected to antibiotic therapy. After three months, a check was made by inspection of the oral cavity, radiography and bone biopsy at the alveolar level. Group A showed a total restitutio ad integrum of the periodontal structures, and in group B still mild gingivitis in 70% of cases and 30% of the state remains unchanged. Numerous experimental studies both in animals and humans have documented that the grafts of porous hydroxyapatite are rapidly invaded by fibrovascular tissue which is subsequently converted into mature lamellar bone tissue by activating osteoblast. Since we acted on the removal of necrotic cementum and rehabilitating the root tissue by polishing without intervention in the ligament but only on anatomical functional interface of cement-blasts, we can connect the positive evolution of the clinical-only component of the cement that could represent this perspective, the only reason that Periodontal Disease become a Cement Disease, while all other clinical elements as nothing more than a clinical pathological accompanying.

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