Search results for: Ignored and underutilized species
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3131

Search results for: Ignored and underutilized species

671 NLRP3-Inflammassome Participates in the Inflammatory Response Induced by Paracoccidioides brasiliensis

Authors: Eduardo Kanagushiku Pereira, Frank Gregory Cavalcante da Silva, Barbara Soares Gonçalves, Ana Lúcia Bergamasco Galastri, Ronei Luciano Mamoni

Abstract:

The inflammatory response initiates after the recognition of pathogens by receptors expressed by innate immune cells. Among these receptors, the NLRP3 was associated with the recognition of pathogenic fungi in experimental models. NLRP3 operates forming a multiproteic complex called inflammasome, which actives caspase-1, responsible for the production of the inflammatory cytokines IL-1beta and IL-18. In this study, we aimed to investigate the involvement of NLRP3 in the inflammatory response elicited in macrophages against Paracoccidioides brasiliensis (Pb), the etiologic agent of PCM. Macrophages were differentiated from THP-1 cells by treatment with phorbol-myristate-acetate. Following differentiation, macrophages were stimulated by Pb yeast cells for 24 hours, after previous treatment with specific NLRP3 (3,4-methylenedioxy-beta-nitrostyrene) and/or caspase-1 (VX-765) inhibitors, or specific inhibitors of pathways involved in NLRP3 activation such as: Reactive Oxigen Species (ROS) production (N-Acetyl-L-cysteine), K+ efflux (Glibenclamide) or phagossome acidification (Bafilomycin). Quantification of IL-1beta and IL-18 in supernatants was performed by ELISA. Our results showed that the production of IL-1beta and IL-18 by THP-1-derived-macrophages stimulated with Pb yeast cells was dependent on NLRP3 and caspase-1 activation, once the presence of their specific inhibitors diminished the production of these cytokines. Furthermore, we found that the major pathways involved in NLRP3 activation, after Pb recognition, were dependent on ROS production and K+ efflux. In conclusion, our results showed that NLRP3 participates in the recognition of Pb yeast cells by macrophages, leading to the activation of the NLRP3-inflammasome and production of IL-1beta and IL-18. Together, these cytokines can induce an inflammatory response against P. brasiliensis, essential for the establishment of the initial inflammatory response and for the development of the subsequent acquired immune response.

Keywords: inflammation, IL-1beta, IL-18, NLRP3, Paracoccidioidomycosis

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670 Insights into Insect Vectors: Liberibacter Interactions

Authors: Murad Ghanim

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The citrus greening disease, also known as Huanglongbing, caused by the phloem-limited bacterium Candidatus Liberibacter asiaticus (CLas) has resulted in tremendous losses and the death of millions of citrus trees worldwide. CLas is transmitted by the Asian citrus psyllid (ACP) Diaphorina citri. The closely-related bacterium Candidatus Liberibacter solanacearum (CLso), which is associated with vegetative disorders in carrots and the zebra chips disease in potatoes, is transmitted by other psyllid species including Bactericera trigonica in carrots and B. ckockerelli in potatoes. Chemical sprays are currently the prevailing method for managing these diseases for limiting psyllid populations; however, they are limited in their effectiveness. A promising approach to prevent the transmission of these pathogens is to interfere with the vector-pathogen interactions, but our understanding of these processes is very limited. CLas induces changes in the nuclear architecture in the midgut of ACP and activates programmed cell death (apoptosis) in this organ. Strikingly, CLso displayed an opposite effect in the gut of B. trigonica, showing limited apoptosis, but widespread necrosis. Electron and fluorescent microscopy further showed that CLas induced the formation of Endoplasmic reticulum (ER) inclusion- and replication-like bodies, in which it increases and multiplies. ER involvement in bacterial replication is hypothesized to be the first stage of an immune response leading to the apoptotic and necrotic responses. ER exploitation and the subsequent events that lead to these cellular and stress responses might activate a cascade of molecular responses ending up with apoptosis and necrosis. Understanding the molecular interactions that underlay the necrotic/apoptotic responses to the bacteria will increase our knowledge of ACP-CLas, and BT-CLso interactions, and will set the foundation for developing novel, and efficient strategies to disturb these interactions and inhibit the transmission.

Keywords: Liberibacter, psyllid, transmission, apoptosis, necrosis

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669 Morphological and Molecular Analysis of Selected Fast-Growing Blue Swimming Crab (Portunus pelagicus) in South of Sulawesi

Authors: Yushinta Fujaya, Andi Ivo Asphama, Andi Parenrengi, Andi Tenriulo

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Blue Swimming crab (Portunus pelagicus) is an important commercial species throughout the subtropical waters and as such constitutes part of the fisheries resources. Data are lacking on the morphological variations of selected fast-growing crabs reared in a pond. This study aimed to analyze the morphological and molecular character of a selected fast-growing crab reared in ponds in South of Sulawesi. The crab seeds were obtained from local fish-trap and hatchery. A study on the growth was carried out in the population of crabs. The dimensions analyzed were carapace width (CW) measured after 3 months of grow out. Morphological character states were examined based on the pattern of spots on the carapace. Molecular analysis was performed using RAPD (Random Amplified Polymorphic DNA). Genetic distance was analysed using TFPGA (Tools for Population Genetic Analyses) version 1.3. The results showed that there were variations in the growth of crabs. These crabs clustered morphologically into three quite distinct groups. The crab with white spots irregularly spread over its carapace was the largest size while the crab with large white spots scattered over the carapace was the smaller size (3%). The crab with small white spots scattered over the carapace was the smallest size found in this study. Molecular analysis showed that there are morphologically and genetically different between groups of crabs. Genetic distances among crabs ranged from 0.1527 to 0.5856. Thus, this study provides information the use of white spots pattern over carapace as indicators to identify the type of blue swimming crabs.

Keywords: crab, portunus pelagicus, morphology, RAPD, Carapace

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668 Coumestrol Induced Apoptosis in Breast Cancer MCF-7 Cells via Redox Cycling of Copper and ROS Generation: Implications of Copper Chelation Strategy in Cancer Treatment

Authors: Atif Zafar Khan, Swarnendra Singh, Imrana Naseem

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Breast cancer is one of the most frequent malignancies in women worldwide and a leading cause of cancer-related deaths among women. Therefore, there is a need to identify new chemotherapeutic strategies for cancer treatment. Unlike normal cells, cancer cells contain elevated copper levels which play an integral role in angiogenesis. Copper is an important metal ion associated with the chromatin DNA, particularly with guanine. Thus, targeting copper via copper-specific chelators in cancer cells can serve as effective anticancer strategy. Keeping in view these facts, we evaluated the anticancer activity and copper-dependent cytotoxic effect of coumestrol (phytoestrogen in soybean products) in breast cancer MCF-7 cells. Coumestrol inhibited proliferation and induced apoptosis in MCF-7 cells, which was prevented by copper chelator neocuproine and ROS scavengers. Coumestrol treatment induced ROS generation coupled to DNA fragmentation, up-regulation of p53/p21, cell cycle arrest at G1/S phase, mitochondrial membrane depolarization and caspases 9/3 activation. All these effects were suppressed by ROS scavengers and neocuproine. These results suggest that coumestrol targets elevated copper for redox cycling to generate ROS leading to DNA fragmentation. DNA damage leads to p53 up-regulation which directs the cell cycle arrest at G1/S phase and promotes caspase-dependent apoptosis of MCF-7 cells. In conclusion, coumestrol induces pro-oxidant cell death by chelating cellular copper to produce copper-coumestrol complexes that engages in redox cycling in breast cancer cells. Thus, targeting elevated copper levels might be a potential therapeutic strategy for selective cytotoxic action against malignant cells.

Keywords: apoptosis, breast cancer, copper chelation, coumestrol, reactive oxygens species, redox cycling

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667 Effects of IMUNO-2865® as Immune Supplement for the Aquaculture Industry

Authors: Ivan Zupan, Tomislav Saric, Suzana Tkalcic

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IMUNO-2865® is a commercially available, β–glucan based, natural hemicellulose compound with proven immunostimulative properties in people, domestic and some aquatic animals. During the experimental feeding trial with IMUNO-2865® in juvenile wild-caught chub under laboratory conditions, supplementation resulted in overall higher growth performance for all experimental groups regardless of the concentration of the added compound. The maximum, 5% concentration of the supplement, resulted in highest weight gain and calculated specific growth rate. In sea bream, as economically most important species in the Mediterranean aquaculture, significant increases in numbers of monocytes and heterophils were observed in the group supplemented with 2.5 % of IMUNO-2865® in the feed. An overall increase of erythrocytes was noted by the end of the experiment, although with variable distribution among groups. Blood Ca++ levels, total proteins, and total NH₃ were significantly higher after 60 days of feeding in all treatment groups compared to the control and remained elevated in the treated group following the secession of supplementation. Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and serum paraoxonase PON1 (U/L) showed similar trends. All these parameters are playing a significant role in either oxygen supplementation of tissues, or anabolic and catabolic processes that on molecular levels contribute to the overall health and immune-building capacity of cells and tissues. The complete lack of mortality in sea bream and presented increases in cellular, biochemical and oxidative stress parameters in the blood suggest that the IMUNO-2865® represents a safe dietary supplement for in aquaculture, with an overall positive and potentially immunostimulative effect on farmed fish.

Keywords: IMUNO-2865®, β–glucans, Mediterranean aquaculture, fish imunnostimulans

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666 Unfolding Global Biodiversity Patterns of Marine Planktonic Diatom Communities across the World's Oceans

Authors: Shruti Malviya, Chris Bowler

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Analysis of microbial eukaryotic diversity is fundamental to understanding ecosystems’ structure, biology, and ecology. Diatoms (Stramenopiles, Bacillariophyceae) are one of the most diverse and ecologically prominent groups of phytoplankton. This study was performed to enhance the understanding of global biodiversity patterns and structure of planktonic diatom communities across the world's oceans. We used the metabarcoding data set generated from the biological samples and associated environmental data collected during the Tara Oceans (2009-2013) global circumnavigation covering all major oceanic provinces. A total of ~18 million diatom V9-18S rDNA tags from 126 sampling stations, constituting 631 size-fractionated plankton communities were generated. Using ~250,000 unique diatom metabarcodes, the global diatom distribution and diversity across size classes, genus and ecological niches was assessed. Notably, our analysis revealed: (i) a new estimate of the total number of planktonic diatom species, (ii) a considerable unknown diversity and exceptionally high diversity in the open ocean, and (iii) complex diversity patterns across oceanic provinces. Also, co-occurrence of several ribotypes in locations separated by great geographic distances (equatorial stations) demonstrated a widespread but not ubiquitous distribution. This work provides a comprehensive perspective on diatom distribution and diversity in the world’s oceans and elaborates interconnections between associated theories and underlying drivers. It shows how meta-barcoding approaches can provide a framework to investigate environmental diversity at a global scale, which is deemed as an essential step in answering various ecological research questions. Consequently, this work also provides a reference point to explore how microbial communities will respond to environmental conditions.

Keywords: diatoms, Tara Oceans, biodiversity, metabarcoding

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665 Identification and Characterization of Oil-Degrading Bacteria from Crude Oil-Contaminated Desert Soil in Northeastern Jordan

Authors: Mohammad Aladwan, Adelia Skripova

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Bioremediation aspects of crude oil-polluted fields can be achieved by isolation and identification of bacterial species from oil-contaminated soil in order to choose the most active isolates and increase the strength of others. In this study, oil-degrading bacteria were isolated and identified from oil-contaminated soil samples in northeastern Jordan. The bacterial growth count (CFU/g) was between 1.06×10⁵ and 0.75×10⁹. Eighty-two bacterial isolates were characterized by their morphology and biochemical tests. The identified bacterial genera included: Klebsiella, Staphylococcus, Citrobacter, Lactobacillus, Alcaligenes, Pseudomonas, Hafnia, Micrococcus, Rhodococcus, Serratia, Enterobacter, Bacillus, Salmonella, Mycobacterium, Corynebacterium, and Acetobacter. Molecular identification of a universal primer 16S rDNA gene was used to identify four bacterial isolates: Microbacterium esteraromaticum strain L20, Pseudomonas stutzeri strain 13636M, Klebsilla pneumoniae, and uncultured Klebsilla sp., known as new strains. Our results indicate that their specific oil-degrading bacteria isolates might have a high strength of oil degradation from oil-contaminated sites. Staphylococcus intermedius (75%), Corynebacterium xerosis (75%), and Pseudomonas fluorescens (50%) showed a high growth rate on different types of hydrocarbons, such as crude oil, toluene, naphthalene, and hexane. In addition, monooxygenase and catechol 2,3-dioxygenase were detected in 17 bacterial isolates, indicating their superior hydrocarbon degradation potential. Total petroleum hydrocarbons were analyzed using gas chromatography for soil samples. Soil samples M5, M7, and M8 showed the highest levels (43,645, 47,805, and 45,991 ppm, respectively), and M4 had the lowest level (7,514 ppm). All soil samples were analyzed for heavy metal contamination (Cu, Cd, Mn, Zn, and Pb). Site M7 contains the highest levels of Cu, Mn, and Pb, while Site M8 contains the highest levels of Mn and Zn. In the future, these isolates of bacteria can be used for the cleanup of oil-contaminated soil.

Keywords: bioremediation, 16S rDNA gene, oil-degrading bacteria, hydrocarbons

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664 Screening and Evaluation of Plant Growth Promoting Rhizobacteria of Wheat/Faba Bean for Increasing Productivity and Yield

Authors: Yasir Arafat, Asma Shah, Hua Shao

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Background and Aims: Legume/cereal intercropping is used worldwide for enhancement in biomass and yield of cereal crops. However, because of intercropping, the belowground biological and chemical interactions and their effect on physiological parameters and yield of crops are limited. Methods: Wheat faba bean (WF) intercropping was designed to understand the underlying changes in the soil's chemical environment, soil microbial communities, and effect on growth and yield parameters. Experimental plots were established as having no root partition (NRP), semi-root partition (SRP), complete root partition (CRP), and their sole cropping (CK). Low molecular weight organic acids (LMWOAs) were determined by GC-MS, and high throughput sequencing of the 16S rRNA gene was carried out to screen microbial structure and composition in different root partitions of the WF intercropping system. Results: We show that intercropping induced a shift in the relative abundance of some genera of plant growth promoting rhizobacteria (PGPR) such as Allorhizobium, Neorhizobium, Pararhizobium, and Rhizobium species and resulted in better growth and yield performance of wheat. Moreover, as the plant's distance of wheat from faba beans decreased, the diversity of microbes increased, and a positive effect was observed on physiological traits and crop yield. Furthermore, an abundance and positive correlations of palmitic acid, arachidic acid, stearic acid, and 9-Octadecenoic with PGPR were recorded in the root zone of WF intercropping, which can play an important role in this facilitative mechanism of enhancing growth and yield of cereals. Conclusion: The two treatments clearly affected soil microbial and chemical composition, which can be reflected in growth and yield enhancement.

Keywords: intercropping, microbial community, LMWOAs, PGPR, soil chemical environment

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663 Kinetics and Mechanism Study of Photocatalytic Degradation Using Heterojunction Semiconductors

Authors: Ksenija Milošević, Davor Lončarević, Tihana Mudrinić, Jasmina Dostanić

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Heterogeneous photocatalytic processes have gained growing interest as an efficient method to generate hydrogen by using clean energy sources and degrading various organic pollutants. The main obstacles that restrict efficient photoactivity are narrow light-response range and high rates of charge carrier recombination. The formation of heterojunction by combining a semiconductor with low VB and a semiconductor with high CB and a suitable band gap was found to be an efficient method to prepare more sensible materials with improved charge separation, appropriate oxidation and reduction ability, and enhanced visible-light harvesting. In our research, various binary heterojunction systems based on the wide-band gap (TiO₂) and narrow bandgap (g-C₃N₄, CuO, and Co₂O₃) photocatalyst were studied. The morphology, optical, and electrochemical properties of the photocatalysts were analyzed by X-ray diffraction (XRD), scanning electron microscopy (FE-SEM), N₂ physisorption, diffuse reflectance measurements (DRS), and Mott-Schottky analysis. The photocatalytic performance of the synthesized catalysts was tested in single and simultaneous systems. The synthesized photocatalysts displayed good adsorption capacity and enhanced visible-light photocatalytic performance. The mutual interactions of pollutants on their adsorption and degradation efficiency were investigated. The interfacial connection between photocatalyst constituents and the mechanism of the transport pathway of photogenerated charge species was discussed. A radical scavenger study revealed the interaction mechanisms of the photocatalyst constituents in single and multiple pollutant systems under solar and visible light irradiation, indicating the type of heterojunction system (Z scheme or type II).

Keywords: bandgap alignment, heterojunction, photocatalysis, reaction mechanism

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662 Nanoparticle Induced Neurotoxicity Mediated by Mitochondria

Authors: Nandini Nalika, Suhel Parvez

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Nanotechnology has emerged to play a vital role in developing all through the industrial world with an immense production of nanomaterials including nanoparticles (NPs). Many toxicological studies have confirmed that due to unique small size and physico-chemical properties of NPs (1-100nm), they can be potentially hazardous. Metallic NPs of small size have been shown to induce higher levels of cellular oxidative stress and can easily pass through the Blood Brain Barrier (BBB) and significantly accumulate in brain. With the wide applications of titanium dioxide nanoparticles (TNPs) in day-to-day life in form of cosmetics, paints, sterilisation and so on, there is growing concern regarding the deleterious effects of TNPs on central nervous system and mitochondria appear to be important cellular organelles targeted to the pro-oxidative effects of NPs and an important source that contribute significantly for the production of reactive oxygen species after some toxicity or an injury. The aim of our study was to elucidate the effect of TNPs in anatase form with different concentrations (5-50 µg/ml) following with various oxidative stress markers in isolated brain mitochondria as an in vitro model. Oxidative stress was determined by measuring the different oxidative stress markers like lipid peroxidation as well as the protein carbonyl content which was found to be significantly increased. Reduced glutathione content and major glutathione metabolizing enzymes were also modulated signifying the role of glutathione redox cycle in the pathophysiology of TNPs. The study also includes the mitochondrial enzymes (Complex 1, Complex II, complex IV, Complex V ) and the enzymes showed toxicity in a relatively short time due to the effect of TNPs. The study provide a range of concentration that were toxic to the neuronal cells and data pointing to a general toxicity in brain mitochondria by TNPs, therefore, it is in need to consider the proper utilization of NPs in the environment.

Keywords: mitochondria, nanoparticles, brain, in vitro

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661 Glyco-Conjugated Gold Nanorods Based Biosensor for Optical Detection and Photothermal Ablation of Food Borne Bacteria

Authors: Shimayali Kaushal, Nitesh Priyadarshi, Nitin Kumar Singhal

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Food borne bacterial species have been identified as major pathogens in most of the severe pathogen-related diseases among humans which result in great loss to human health and food industry. Conventional methods like plating and enzyme-linked immune sorbent assay (ELISA) are time-consuming, laborious and require specialized instruments. Nanotechnology has emerged as a great field in case of rapid detection of pathogens in recent years. The AuNRs material has good electro-optical properties due to its larger light absorption band and scattering in surface plasmon resonance wavelength regions. By exploiting the sugar-based adhesion properties of microorganism, we can use the glycoconjugates capped gold nanorods as a potential nanobiosensor to detect the foodborne pathogen. In the present study, polyethylene glycol (PEG) coated gold nanorods (AuNRs) were prepared and functionalized with different types of carbohydrates and further characterized by UV-Visible spectrophotometry, dynamic light scattering (DLS), transmission electron microscopy (TEM). The reactivity of above said nano-biosensor was probed by lectin binding assay and also by different strains of foodborne bacteria by using spectrophotometric and microscopic techniques. Due to the specific interaction of probe with foodborne bacteria (Escherichia coli, Pseudomonas aeruginosa), our nanoprobe has shown significant and selective ablation of targeted bacteria. Our findings suggest that our nanoprobe can be an ideal candidate for selective optical detection of food pathogens and can reduce loss to the food industry.

Keywords: glyco-conjugates, gold nanorods, nanobiosensor, nanoprobe

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660 Anti-Oxidant and Anti-Cancer Activity of Helix aspersa Aqueous Extract

Authors: Ibtissem El Ouar, Cornelia Braicu, Dalila Naimi, Alexendru Irimie, Ioana Berindan-Neagoe

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Helix aspersa, 'the garden snail' is a big land snail widely found in the Mediterranean countries, it is one of the most consumed species in the west of Algeria. It is commonly used in zootherapy to purify blood and to treat cardiovascular diseases and liver problems. The aim of our study is to investigate, the antitumor activity of an aqueous extract from Helix aspersa prepared by the traditional method on Hs578T; a triple negative breast cancer cell line. Firstly, the free radical scavenging activity of H. aspersa extract was assessed by measuring its capability for scavenging the radical 2,2-diphenyl-1-picrylhydrazyl (DPPH), as well as its ability to reduce ferric ion by the FRAP assay (ferric reducing ability). The cytotoxic effect of H. aspersa extract against Hs578T cells was evaluated by the MTT test (3-(4,5- dimethylthiazl-2-yl)-2,5- diphenyltetrazolium bromide)) while the mode of cell death induced by the extract has been determined by fluorescence microscopy using acredine orange/ethidium bromide (AO/EB) probe. The level of TNFα has also measured in cell medium by ELISA method. The results suggest that H. aspersa extract has an antioxidant activity, especially at high concentrations, it can reduce DPPH radical and ferric ion. The MTT test shows that H. aspersa extract has a great cytotoxic effect against breast cancer cells, the IC50 value correspond of the dilution 1% of the crude extract. Moreover, the AO/EB staining shows that TNFα induced necrosis is the main form of cell death induced by the extract. In conclusion, the present study may open new perspectives in the search for new natural anticancer drugs.

Keywords: breast cancer, Helix aspersa, Hs578t cell line, necrosis

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659 Variation of Fertility-Related Traits in Italian Tomato Landraces under Mild Heat Stress

Authors: Maurizio E. Picarella, Ludovica Fumelli, Francesca Siligato, Andrea Mazzucato

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Studies on reproductive dynamics in crops subjected to heat stress are crucial to breed more tolerant cultivars. In tomato, cultivars, breeding lines, and wild species have been thoroughly evaluated for the response to heat stress in several studies. Here, we address the reaction to temperature stress in a panel of selected landraces representing genotypes cultivated before the advent of professional varieties that usually show high adaptation to local environments. We adopted an experimental design with two open field trials, where transplanting was spaced by one month. In the second field, plants were thus subjected to mild stress with natural temperature fluctuations. The genotypes showed wide variation for both vegetative (plant height) and reproductive (stigma exsertion, pollen viability, number of flowers per inflorescence, and fruit set) traits. On average, all traits were affected by heat conditions; except for the number of flowers per inflorescence, the “G*E” interaction was always significant. In agreement with studies based on different materials, estimated broad sense heritability was high for plant height, stigma exsertion, and pollen viability and low for the number of flowers per inflorescence and fruit set. Despite the interaction, traits recorded in control and in heat conditions were positively correlated. The first two principal components estimated by multivariate analysis explained more than 50% of the total variability. The study indicated that landraces present a wide variability for the response of reproductive traits to temperature stress and that such variability could be very informative to dissect the traits with higher heritability and identify new QTL useful for breeding more resilient varieties.

Keywords: fruit set, heat stress, solanum lycopersicum L., style exsertion, tomato

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658 Genetic Assessment of The Managed Gharial Population In The Girwa River, India

Authors: Surya Prasad Sharma, Suyash Katdare, Syed Ainul Hussain

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Human-induced factors contributed to the population decline of crocodylians in India which became evident by the mid-20th century when authorities forewarned the extinction risk for the crocodile and proposed regulation in the crocodile trade. The proposed action led to the enactment of national and international wildlife regulations to prohibit the trade-in of crocodile skins and parts. Subsequently, conservation translocation programs were initiated to restore the species in the wild through a 'head-start' approach. In India, the crocodile conservation program, which began in the early 1970s, has been one of India's longest-running conservation initiatives. The gharial (Gavialis gangeticus) population has benefitted, and the gharial number increased rapidly owing to these efforts. The immediate risk of extinction was averted as the gharial has recovered due to decades-long cumulative conservation efforts, the consideration of the genetic for monitoring the recovery of the recovered populations is still lacking. Hence, we assessed the genetic diversity of the Girwa gharial population in India using six polymorphic nuclear microsatellites loci and mitochondrial control region. The number of alleles per loci ranged between 2 to 5, and the allelic richness (Ar) was 2.67 ± 0.49, and the observed (Ho) and expected (He) heterozygosities were 0.42 ± 0.08 and 0.42 ± 0.09, respectively. The M-ratio yielded a value of (0.41 ± 0.16) lower than critical M, suggesting a genetic bottleneck in the Girwa population. We observed more mitochondrial control region haplotypes in the Girwa population than previously reported in the largest gharial population in the Chambal River. Overall, our study indicates that genetic diversity remains low despite the recovery in the Girwa population. Hence, we recommend a range-wide genetic assessment of gharial populations using high-throughput techniques to identify the source population and plan future translocation programs.

Keywords: conservation translocation, recovery, crocodile, bottleneck

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657 Effect of Mixture of Flaxseed and Pumpkin Seeds Powder on Hypercholesterolemia

Authors: Zahra Ashraf

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Flax and pumpkin seeds are a rich source of unsaturated fatty acids, antioxidants and fiber, known to have anti-atherogenic properties. Hypercholesterolemia is a state characterized by the elevated level of cholesterol in the blood. This research was designed to study the effect of flax and pumpkin seeds powder mixture on hypercholesterolemia and body weight. Rat’s species were selected as human representative. Thirty male albino rats were divided into three groups: a control group, a CD-chol group (control diet+cholesterol) fed with 1.5% cholesterol and FP-chol group (flaxseed and pumpkin seed powder+ cholesterol) fed with 1.5% cholesterol. Flax and pumpkin seed powder mixed at proportion of (5/1) (omega-3 and omega-6). Blood samples were collected to examine lipid profile and body weight was also measured. Thus the data was subjected to analysis of variance. In CD-chol group, body weight, total cholesterol TC, triacylglycerides TG in plasma, plasma LDL-C, ratio significantly increased with a decrease in plasma HDL (good cholesterol). In FP-chol group lipid parameters and body weights were decreased significantly with an increase in HDL and decrease in LDL (bad cholesterol). The mean values of body weight, total cholesterol, triglycerides, low density lipoprotein and high density lipoproteins in FP-chol group were 240.66±11.35g, 59.60±2.20mg/dl, 50.20±1.79 mg/dl, 36.20±1.62mg/dl, 36.40±2.20 mg/dl, respectively. Flaxseed and pumpkin seeds powder mixture showed reduction in body weight, serum cholesterol, low density lipoprotein and triglycerides. While significant increase was shown in high density lipoproteins when given to hypercholesterolemic rats. Our results suggested that flax and pumpkin seed mixture has hypocholesterolemic effects which were probably mediated by polyunsaturated fatty acids (omega-3 and omega-6) present in seed mixture.

Keywords: hypercolesterolemia, omega 3 and omega 6 fatty acids, cardiovascular diseases

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656 Biotic Potential of Different Densities of Aphid Parasitoids, Diaeretiella rapae (Hymenoptera: Braconidae: Aphidiinae) Feeding on Brevicoryne brassicae

Authors: Muhammad Anjum Aqueel, Muhammad Jaffar Hussain, Abu Bakar Muhammad Raza

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Diaeretiella rapae (M’Intosh) attack most of the aphid species. However, it is specialized in feeding on crucifer aphid, Brevicoryne brassicae. Biological potential of parasitoid is its density-dependency due to sharing of limited resources in few cases. The present study was carried out to check the biotic potential of D. rapae at its different densities (1, 2, 4, 8 and 10 pairs) on fixed number of B. brassicae (100 in number) as a host. The present study was performed under laboratory conditions (25 ± 2 ºC temperature and 65-70 % R.H.). Different biological parameters for parasitoid (e.g. percent parasitism, adult emergence, adult longevity and per pair parasitism) were evaluated to check its biotic potential. The present findings showed that maximum parasitism (43.09 % ± 0.63) was observed in highest density (10 pairs) and minimum parasitism (16.59 % ± 1.28) in lowest density (1 pair) of the parasitoid. Maximum adult emergence (80.31 % ± 1.33) was observed in highest density (10 pairs) and minimum parasitism (45.99 % ± 1.27) in lowest density (1 pair) of the parasitoid. In the case of adult longevity, highest (8.2 days ± 0.38) and lowest (6 days ± 0.32) longevity were observed in lowest (1 pair) and highest (10 pairs) densities of parasitoids respectively. However, per pair parasitism rate decreased with the increase in parasitoid densities due to intra-specific competition, developed between the parasitoids for parasitism. The positive but close relationship was observed between percent parasitism and adult emergence. The increase in parasitoid densities increased the percent parasitism and adult emergence of the parasitoid. So, we conclude that an inter-specific competition negatively affected the efficacy of parasitoids and may reduce the fitness of the emerging parasitoid.

Keywords: Diaeretiella rapae, Parasitoid densities, Percent parasitism, adult emergence

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655 Spectral Mixture Model Applied to Cannabis Parcel Determination

Authors: Levent Basayigit, Sinan Demir, Yusuf Ucar, Burhan Kara

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Many research projects require accurate delineation of the different land cover type of the agricultural area. Especially it is critically important for the definition of specific plants like cannabis. However, the complexity of vegetation stands structure, abundant vegetation species, and the smooth transition between different seconder section stages make vegetation classification difficult when using traditional approaches such as the maximum likelihood classifier. Most of the time, classification distinguishes only between trees/annual or grain. It has been difficult to accurately determine the cannabis mixed with other plants. In this paper, a mixed distribution models approach is applied to classify pure and mix cannabis parcels using Worldview-2 imagery in the Lakes region of Turkey. Five different land use types (i.e. sunflower, maize, bare soil, and cannabis) were identified in the image. A constrained Gaussian mixture discriminant analysis (GMDA) was used to unmix the image. In the study, 255 reflectance ratios derived from spectral signatures of seven bands (Blue-Green-Yellow-Red-Rededge-NIR1-NIR2) were randomly arranged as 80% for training and 20% for test data. Gaussian mixed distribution model approach is proved to be an effective and convenient way to combine very high spatial resolution imagery for distinguishing cannabis vegetation. Based on the overall accuracies of the classification, the Gaussian mixed distribution model was found to be very successful to achieve image classification tasks. This approach is sensitive to capture the illegal cannabis planting areas in the large plain. This approach can also be used for monitoring and determination with spectral reflections in illegal cannabis planting areas.

Keywords: Gaussian mixture discriminant analysis, spectral mixture model, Worldview-2, land parcels

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654 Interaction between the Rio Conventions on Climate and Biodiversity: Analysis of the Integration of Ecosystem-Based Approaches and Nature-Based Solutions into the UNFCCC

Authors: Dieudonne Mevono Mvogo

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The Intergovernmental Platform on Biodiversity and Ecosystem Services (IPBES)-Intergovernmental Panel on Climate Change (IPCC) co-sponsored workshop report suggests that climate change and biodiversity loss are two of the most pressing issues of the Anthropocene. Research establishes the interconnection between climate change and biodiversity. On the one hand, the impact of climate change on biodiversity loss – 14 % over the past century – is projected to surpass other threats – land and sea use 34 % and direct exploitation of species 23 % – during the 21st century. Response measures to climate change also affect biodiversity negatively or positively. On the other hand, actions to halt or reverse biodiversity loss can enhance land and ocean capacity for carbon sequestration. These actions can also promote adaptation by ensuring adaptive capacity. This systemic interaction between climate change and biodiversity affects the human quality of life. The United Nations Secretariat's report entitled 'Gaps in international environmental law and environment-related instruments: towards a global pact for the environment,' released in 2018, states that cooperation and mutual support among agreements dealing with climate change, the protection of the marine environment, freshwater resources and hazardous waste are indispensable for the effective implementation of the Convention on the Biological Diversity (CBD). Since biodiversity is being lost at an alarming rate, this study aims to evaluate the cooperative framework for the coherence and coordination between climate change and biodiversity regimes to provide co-benefits for climate and biodiversity crises. It questions the potential improvement regarding integrating ecosystem-based approaches and nature-based solutions – promoted by the CBD – into the United Nations Framework Convention on Climate Change (UNFCCC).

Keywords: rio conventions, climate change, biodiversity, cooperative framework, ecosystem-based approaches, nature-based solutions

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653 Economic Important of Manta Ray Watching Tourism in Dampier Strait, Raja Ampat, West Papua, Indonesia

Authors: Maulita Sari Hani, Abraham B. Sianipar, Jamaluddin Jompa, Natsir Nessa, Alan T. White

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Manta ray is an icon for tourism in Raja Ampat. The tourist volume has been increased for the past ten years which up to approximately 23,000 tourists in 2017. Since 2013, Conservation International Indonesia deployed satellite and acoustic tags on manta ray in Dampier strait to track the species and identify the aggregation areas. These findings encourage the government and the local community to boost conservation through the management of marine protected areas for tourism purposes. Community in Dampier strait including the village of Arborek, Kurkapa, Kapisawar, and Sawingray involved in variety of small scale tourism business including homestay, dive shop, tour operator, and crafts. Working groups of related local businesses were established to support the local community and to ensure the sustainability of the economic viability and environmental sustainability. In order to analyze the economic benefits of manta ray tourism, this study was conducted to identify the number of local business in Dampier Strait and the economic impacts in terms of local finance security, social, humanity, individual, and physical assets. The results of this study identify 30 homestays, 2 dive shops, 10 tour operators, 30 women involved in crafts, and about 50 villagers worked for dive resorts. In addition to community assets, we confirmed the welfare of community has been improved in terms of food security, households, education for children, savings, and health insurance.

Keywords: marine wildlife tourism, elasmobranch, conservation, ecotourism, co-management, economic viability, environmental sustainability

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652 Economic Assessment of the Fish Solar Tent Dryers

Authors: Collen Kawiya

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In an effort of reducing post-harvest losses and improving the supply of quality fish products in Malawi, the fish solar tent dryers have been designed in the southern part of Lake Malawi for processing small fish species under the project of Cultivate Africa’s Future (CultiAF). This study was done to promote the adoption of the fish solar tent dryers by the many small scale fish processors in Malawi through the assessment of the economic viability of these dryers. With the use of the project’s baseline survey data, a business model for a constructed ‘ready for use’ solar tent dryer was developed where investment appraisal techniques were calculated in addition with the sensitivity analysis. The study also conducted a risk analysis through the use of the Monte Carlo simulation technique and a probabilistic net present value was found. The investment appraisal results showed that the net present value was US$8,756.85, the internal rate of return was 62% higher than the 16.32% cost of capital and the payback period was 1.64 years. The sensitivity analysis results showed that only two input variables influenced the fish solar dryer investment’s net present value. These are the dried fish selling prices that were correlating positively with the net present value and the fresh fish buying prices that were negatively correlating with the net present value. Risk analysis results showed that the chances that fish processors will make a loss from this type of investment are 17.56%. It was also observed that there exist only a 0.20 probability of experiencing a negative net present value from this type of investment. Lastly, the study found that the net present value of the fish solar tent dryer’s investment is still robust in spite of any changes in the levels of investors risk preferences. With these results, it is concluded that the fish solar tent dryers in Malawi are an economically viable investment because they are able to improve the returns in the fish processing activity. As such, fish processors need to adopt them by investing their money to construct and use them.

Keywords: investment appraisal, risk analysis, sensitivity analysis, solar tent drying

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651 The Evaluation of Adjuvant Effects of CD154 in a Subunit Vaccine against Classical Swine Fever Virus

Authors: Yu-Chieh Chen, Li-Yun Wang, Chi-Chih Chen, Huy Hùng Đào, Ya-Mei Chen, Ming-Chu Cheng, Wen-Bin Chung, Hso-Chi Chaung, Guan-Ming Ke

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Many recent researches have demonstrated that CD154, a protein primarily expressed on activated T cell molecules, has potentially acted as a molecular adjuvant to improve the immunogenicity of subunit vaccines against viral infections. Classical swine fever (CSF) affects the swine industry worldwide that is one of the most devastating and highly contagious pig diseases. It is listed by the World Organization for Animal Health (OIE) as an infectious animal disease that must be reported. Although pigs vaccinated with subunit vaccines can be differentially diagnosed from those infected animals, subunit vaccines usually need adjuvants to enhance and elicit immune responses. In this study, CD154 was linked with CSFV E2 sequences and then expressed in CHO cells to produce the fusion protein as E2-CD154. The porcine specific CpG adjuvant was also used in one of the formulations. The specific pathogen-free pigs (SPF) at the age of 4-week-old were randomly separated into four groups, vaccinated with E2-CpG, E2-CD154, E2-CD154-CpG or the commercial Bayovac® CSF-E2 vaccine and boosted two weeks after primary vaccination. The results showed that the percentages of CD4+ and CD4+IL2+ in peripheral blood mononuclear cells (PBMC) in E2-CD154 vaccinated piglets seven days after primary vaccination were gained by 1-5% relative to the control group. In addition, the percentages of CD4+IFNγ+ T cells had slightly edged up 0.1-0.3% compared with the control group. Also, increased E2-specific IFNγ levels had edged up CD4+CD8+ T cells found in E2-CD154 and E2-CD154-CpG groups, particularly in the E2-CD154-CpG group. These results implicate that CD154 may enhance cellular immunity and synergistically act with species-specific CpG adjuvant as a dual-phase adjuvant. Therefore, the CD154 may be beneficial as a promising adjuvant in subunit vaccines.

Keywords: CD154, CpG adjuvant, cellular immunity, subunit vaccine, pig

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650 Optimization and Kinetic Analysis of the Enzymatic Hydrolysis of Oil Palm Empty Fruit Bunch To Xylose Using Crude Xylanase from Trichoderma Viride ITB CC L.67

Authors: Efri Mardawati, Ronny Purwadi, Made Tri Ari Penia Kresnowati, Tjandra Setiadi

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EFB are mainly composed of cellulose (≈ 43%), hemicellulose (≈ 23%) and lignin (≈20%). The palm oil empty fruit bunches (EFB) is the lignosellulosic waste from crude palm oil industries mainly compose of (≈ 43%), hemicellulose (≈ 23%) and lignin (≈20%). Xylan, a polymer made of pentose sugar xylose and the most abundant component of hemicellulose in plant cell wall. Further xylose can be used as a raw material for production of a wide variety of chemicals such as xylitol, which is extensively used in food, pharmaceutical and thin coating applications. Currently, xylose is mostly produced from xylan via chemical hydrolysis processes. However, these processes are normally conducted at a high temperature and pressure, which is costly, and the required downstream processes are relatively complex. As an alternative method, enzymatic hydrolysis of xylan to xylose offers an environmentally friendly biotechnological process, which is performed at ambient temperature and pressure with high specificity and at low cost. This process is catalysed by xylanolytic enzymes that can be produced by some fungal species such as Aspergillus niger, Penicillium crysogenum, Tricoderma reseei, etc. Fungal that will be used to produce crude xylanase enzyme in this study is T. Viride ITB CC L.67. It is the purposes of this research to study the influence of pretreatment of EFB for the enzymatic hydrolysis process, optimation of temperature and pH of the hydrolysis process, the influence of substrate and enzyme concentration to the enzymatic hydrolysis process, the dynamics of hydrolysis process and followingly to study the kinetics of this process. Xylose as the product of enzymatic hydrolysis process analyzed by HPLC. The results show that the thermal pretreatment of EFB enhance the enzymatic hydrolysis process. The enzymatic hydrolysis can be well approached by the Michaelis Menten kinetic model, and kinetic parameters are obtained from experimental data.

Keywords: oil palm empty fruit bunches (EFB), xylose, enzymatic hydrolysis, kinetic modelling

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649 Dissolved Black Carbon Accelerates the Photo-Degradation of Polystyrene Microplastics

Authors: Qin Ou, Yanghui Xu, Xintu Wang, Kim Maren Lompe, Gang Liu, Jan Peter Van Der Hoek

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Microplastics (MPs) can undergo the photooxidation process under ultraviolet (UV) exposure, which determines their transformation and fate in environments. The presence of dissolved organic matter (DOM) can interact with MPs and take participate in the photo-degradation of MPs. As an important DOM component, dissolved black carbon (DBC), widely distributed in aquatic environments, can accelerate or inhibit the sunlight-driven photo-transformation of environmental pollutants. However, the role and underlying mechanism of DBC in the photooxidation of MPs are not clear. Herein, the DBC (< 0.45 µm) was extracted from wood biochar and fractionated by molecular weight (i.e., <3 KDa, 3 KDa−30 KDa, 30 KDa−0.45 µm). The effects of DBC chemical composition (i.e., molecular weight and chemical structure) in DBC-mediated photo-transformation of polystyrene (PS) MPs were investigated. The results showed that DBC initially inhibited the photo-degradation of MPs due to light shielding. Under UV exposure for 6−24 h, the presence of 5 mg/L DBC decreased the carbonyl index of MPs compared to the control. This inhibitory effect of DBC was found to decrease with increasing irradiation time. Notably, DBC initially decreased but then increased the hydroxyl index with aging time, suggesting that the role of DBC may shift from inhibition to acceleration. In terms of the different DBC fractions, the results showed that the smallest fraction of DBC (<3 KDa) significantly accelerated the photooxidation of PS MPs since it acted as reactive oxygen species (ROS) generators, especially in promoting the production of ¹O₂ and ³DBC* and •OH. With the increase in molecular weight, the acceleration effect of DBC on the degradation of MPs was decreased due to the increase of light shielding and possible decrease of photosensitization ability. This study thoroughly investigated the critical role of DBC chemical composition in the photooxidation process, which helps to assess the duration of aging and transformation of MPs during long-term weathering in natural waters.

Keywords: microplastics, photo-degradation, dissolved black carbon, molecular weight, photosensitization

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648 Hydrodynamics of Periphyton Biofilters in Recirculating Aquaculture

Authors: Adam N. Bell, Sarina J. Ergas, Michael Nystrom, Nathan P. Brennan, Kevan L. Main

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Integrated Multi-Trophic Aquaculture systems (IMTA) have the potential to improve the sustainability of seafood production, generate organic fertilizer and feed, remove waste discharges and reduce energy use. IMTA can include periphyton biofilters where algae and microbes grow on surfaces, along with caught detritus and amphipods. Periphyton biofilters provide many advantages: nitrification, denitrification, primary production and ecological diversity. The goal of this study was to determine how biofilter hydraulic residence time (τ) effects periphyton biomass production, dissolved oxygen (DO) and nutrient removal. A pilot scale recirculating aquaculture system (RAS) was designed, constructed and operated at different hydraulic residence times (τ= 1, 2, 4, 6, 8 hours per tank). For each τ, a conservative tracer study was conducted to investigate system hydrodynamics. Data on periphyton weights, pH, nitrogen species, phosphorus, temperature and DO were collected. The tracer study for τ =1 hour revealed that the normalized time < τ, indicating short-circuiting. Periphyton biomass production rate was relatively unaffected by τ (R_e<1 for all τ). Average ammonia nitrogen removal was > 75% for all trials. Nitrate and nitrite did not accumulate in the RAS for τ≥4 hours due to enhanced denitrification in anoxic zones. For τ≥4 hours DO concentration was at a maximum of 4 mg L-1 after 14:00, and decreased to 0 mg L-1 during nighttime. At τ=1 hour, the RAS stayed > 2 mg L-1 and DO was more evenly distributed. For the validation trial, the culture tank was stocked with Centropomus undecimalis (common snook) and the system was operated at τ= 1 hr. Preliminary results showed that a RAS with an integrated periphyton biofilter could support fish health with low nutrient concentrations DO > 6 mg L-1.

Keywords: sustainable aquaculture, resource recovery, nitrogen, microalgae, hydrodynamics, integrated multi-trophic aquaculture

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647 Role of Climatic Conditions on Pacific Bluefin Tuna Thunnus orientalis Stock Structure

Authors: Ashneel Ajay Singh, Kazumi Sakuramoto, Naoki Suzuki, Kalla Alok, Nath Paras

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Bluefin (Thunnus orientalis) tuna is one of the most economically valuable tuna species in the world. In recent years the stock has been observed to decline. It is suspected that the stock-recruitment relationship and population structure is influenced by environmental and climatic variables. This study was aimed at investigating the influence of environmental and climatic conditions on the trajectory of the different life stages of the North Pacific bluefin tuna. Exploratory analysis was performed for the North Pacific sea surface temperature (SST) and Pacific Decadal Oscillation (PDO) on the time series of the bluefin tuna cohorts (age-0, 1, 2,…,9, 10+). General Additive Modeling (GAM) was used to reconstruct the recruitment (R) trajectory. The spatial movement of the SST was also monitored from 1953 to 2012 in the distribution area of the bluefin tuna. Exploratory analysis showed significance influence of the North Pacific Sea Surface temperature (SST) and Pacific Decadal Oscillation (PDO) on the time series of the age-0 group. Other age group (1, 2,…,9, 10+) time series did not exhibit any significant correlations. PDO showed most significant relationship in the months of October to December. Although the stock-recruitment relationship is of biological significance, the recruits (age-0) showed poor correlation with the Spawning Stock Biomass (SSB). Indeed the most significant model incorporated the SSB, SST and PDO. The results show that the stock-recruitment relationship of the North Pacific bluefin tuna is multi-dimensional and cannot be adequately explained by the SSB alone. SST and PDO forcing of the population structure is of significant importance and needs to be accounted for when making harvesting plans for bluefin tuna in the North Pacific.

Keywords: pacific bluefin tuna, Thunnus orientalis, cohorts, recruitment, spawning stock biomass, sea surface temperature, pacific decadal oscillation, general additive model

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646 Cytotoxicity thiamethoxam Study on the Hepatopancreas and Its Reversibility under the Effect of Ginger in Helix aspersa

Authors: Samira Bensoltane, Smina Ait Hamlet, Samti Meriem, Semmasel Asma

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Living organisms in the soil are subject to regular fluctuations of abiotic parameters, as well as a chemical contamination of the environment due to human activities. They are subject to multiple stressors they face. The aim of our work was to study the effects of insecticide: thiamethoxam (neonicotinoid), and the potential reversibility of the effects by an antioxidant: ginger on a bioindicator species in ecotoxicology, the land snail Helix aspersa. The effects were studied by a targeted cell approach of evaluating the effect of these molecules on tissue and cellular aspect of hepatopancreas through histological study. Treatment with thiamethoxam concentrations 10, 20, and 40 mg/l shows signs of inflammation even at low concentrations and from the 5th day of treatment. Histological examination of the hepatopancreas of snails treated with thiamethoxam showed significant changes from the lowest concentrations tested , note intertubular connective tissue enlargement, necrosis deferent types of cells (cells with calcium , digestive, excretory) , also damage acini, alteration of the apical membrane and lysis of the basement membrane in a dose- dependent manner. After 10 days of treatment and with 40 mg/l, the same changes were observed with a very advanced degeneration of the wall of the member that could be confused with the cell debris. For cons, the histological study of the hepatopancreas in Helix aspersa treated with ginger for a period of 15 days after stopping treatment with thiamethoxam has shown a partial regeneration of hepatopancreatic tissue snails treated with all concentrations of thiamethoxam and especially in the intertubular connective tissue of the wall and hepatopancreatic digestive tubules. Finally, we can conclude that monitoring the effect of the insecticide thiamethoxam showed significant alterations, however, treatment with ginger shows regeneration of damaged cells themselves much sharper at low concentration (10 mg/L).

Keywords: Helix aspersa, insecticides, thiamethoxam, ginger, hepatopancreas

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645 Treatment of Histopathological Symptoms in N-Nitrosopyrrolidine Induced Changes in Lung Tissue by Isolated Flavonoid from Indigofera tinctoria

Authors: Aastha Agarwal, Veena Sharma

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N-nitrosopyrollidine or NPYR is a tobacco-specific nitrosamine which upon intoxicated causes abnormal production of Reactive Oxygen Species disrupt the endogenous antioxidant system. The study was designed to evaluate the histological changes in lung tissue of Mus musculus in NPYR administered lungs and effect of isolated flavonoid 3,6-dihydroxy-(3’,4’,7’-trimethoxyphenyl)-chromen-4-one-7-glucoside (ITC) from experimental plant Indigofera tinctorial. Post treatment with isolated compound significantly restored the abnormal symptoms and changes in pulmonary tissue. Transverse section of mouse lung in control animals appeared as a thin lace. Histologically, most of the lung was arranged as alveoli which were thin walled structures made up of single layered squamous epithelial cells. In the transverse section of lung at 100 X will clearly show the component of alveoli, surround by a thin layer of connective tissue and blood vessels. Smaller bronchioles were lined by cuboidal epithelial cells while larger bronchioles were lined by ciliated columnar epithelium layer while in NPYR intoxicated lungs signs of vast pulmonary damages and carcinogenesis as alveolar damage, necrosis, DADs or defused alveolar damages hyperplasia, metaplasia, dysplasia and next stage of carcinogenesis were revealed. Treatment with ITC showed the significant positive changes in the lung tissue due to the side hydroxyl and methoxy groups in its structure which help in combating oxidative injuries and give protection from the free radicals generated during the metabolism of NPYR in body. Thus, histopathological analysis confirms the development of the cancerous conditions in the lung tissue in mice model and the protective effects of ITC.

Keywords: flavonoid, histopathology, Indigofera tinctoria, lung

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644 Effect of Co-Infection With Intestinal Parasites on COVID-19 Severity: A Prospective Observational Cohort Study

Authors: Teklay Gebrecherkos, Dawit Wolday, Muhamud Abdulkader

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Background: COVID-19 symptomatology in Africa appears significantly less serious than in the industrialized world. Our hypothesis for this phenomenon, being a different, more activated immune system due to parasite infections contributes to reduced COVID-19 outcome. We investigated this hypothesis in an endemic area in sub sub-saharan Africa. Methods: Ethiopian COVID-19 patients were enrolled and screened for intestinal parasites, between July 2020 and March 2021. The primary outcome was the proportion of patients with severe COVID-19. SARS-CoV-2 infection was confirmed by RT-PCR on samples obtained from nasopharyngeal swabs, while direct microscopic examination, modified Ritchie concentration, and Kato-Katz methods were used to identify parasites and ova from a fresh stool sample. Ordinal logistic regression models were used to estimate the association between parasite infection and COVID-19 severity. Models were adjusted for sex, age, residence, education level, occupation, body mass index, and comorbidities. Data were analyzed using STATA version 14. P-value <0.05 was considered statistically significant. Results: A total of 751 SARS-CoV-2 infected patients were enrolled, of whom 284 (37•8%) had an intestinal parasitic infection. Only 27/255 (10•6%) severe COVID-19 patients were co-infected with intestinal parasites, while 257/496 (51•8%) non-severe COVID-19 patients appeared parasite positive (p<0.0001). Patients co-infected with parasites had lower odds of developing severe COVID-19, with an adjusted odds ratio (AOR) of 0•14 (95% CI 0•09–0•24; p<0•0001) for all parasites, AOR 0•20 ([95% CI 0•11–0•38]; p<0•0001) for protozoa, and AOR 0•13 ([95% CI 0•07–0•26]; p<0•0001) for helminths. When stratified by species, co-infection with Entamoeba spp., Hymenolopis nana, and Schistosoma mansoni implied a lower probability of developing severe COVID-19. There were 11 deaths (1•5%), and all were among patients without parasites (p=0•009). Conclusions: Parasite co-infection is associated with a reduced risk of severe COVID-19 in African patients. Parasite-driven immunomodulatory responses may mute hyper-inflammation associated with severe COVID-19.

Keywords: COVID-19, SARS-COV-2, intestinal parasite, RT-PCR, co-infection

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643 Study of Virus/es Threatening Large Cardamom Cultivation in Sikkim and Darjeeling Hills of Northeast India

Authors: Dharmendra Pratap

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Large Cardamom (Amomum subulatum), family Zingiberaceae is an aromatic spice crop and has rich medicinal value. Large Cardamom is as synonymous to Sikkim as Tea is to Darjeeling. Since Sikkim alone contributes up to 88% of India's large cardamom production which is the world leader by producing over 50% of the global yield. However, the production of large cardamom has declined almost to half since last two decade. The economic losses have been attributed to two viral diseases namely, chirke and Foorkey. Chirke disease is characterized by light and dark green streaks on leaves. The affected leaves exhibit streak mosaic, which gradually coalesce, turn brown and eventually dry up. Excessive sprouting and formation of bushy dwarf clumps at the base of mother plants that gradually die characterize the foorkey disease. In our surveys in Sikkim–Darjeeling hill area during 2012-14, 40-45% of plants were found to be affected with foorkey disease and 10-15% with chirke. Mechanical and aphid transmission study showed banana as an alternate host for both the disease. For molecular identification, total genomic DNA and RNA was isolated from the infected leaf tissues and subjected to Rolling circle amplification (RCA) and RT-PCR respectively. The DNA concatamers produced in the RCA reaction were monomerized by different restriction enzymes and the bands corresponding to ~1 kb genomes were purified and cloned in the respective sites. The nucleotide sequencing results revealed the association of Nanovirus with the foorkey disease of large cardamom. DNA1 showed 74% identity with Replicase gene of FBNYV, DNA2 showed 77% identity with the NSP gene of BBTV and DNA3 showed 74% identity with CP gene of BBTV. The finding suggests the presence of a new species of nanovirus associated with foorkey disease of large cardamom in Sikkim and Darjeeling hills. The details of their epidemiology and other factors would be discussed.

Keywords: RCA, nanovirus, large cardamom, molecular virology and microbiology

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642 Anti-Parasite Targeting with Amino Acid-Capped Nanoparticles Modulates Multiple Cellular Processes in Host

Authors: Oluyomi Stephen Adeyemi, Kentaro Kato

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Toxoplasma gondii is the etiological agent of toxoplasmosis, a common parasitic disease capable of infecting a range of hosts, including nearly one-third of the human population. Current treatment options for toxoplasmosis patients are limited. In consequence, toxoplasmosis represents a large global burden that is further enhanced by the shortcomings of the current therapeutic options. These factors underscore the need for better anti-T. gondii agents and/or new treatment approach. In the present study, we sought to find out whether preparing and capping nanoparticles (NPs) in amino acids, would enhance specificity toward the parasite versus the host cell. The selection of amino acids was premised on the fact that T. gondii is auxotrophic for some amino acids. The amino acid-nanoparticles (amino-NPs) were synthesized, purified and characterized following established protocols. Next, we tested to determine the anti-T. gondii activity of the amino-NPs using in vitro experimental model of infection. Overall, our data show evidence that supports enhanced and excellent selective action against the parasite versus the host cells by amino-NPs. The findings are promising and provide additional support that warrants exploring the prospects of NPs as alternative anti-parasite agents. In addition, the anti-parasite action by amino-NPs indicates that nutritional requirement of parasite may represent a viable target in the development of better alternative anti-parasite agents. Furthermore, data suggest the anti-parasite mechanism of the amino-NPs involves multiple cellular processes including the production of reactive oxygen species (ROS), modulation of hypoxia-inducing factor-1 alpha (HIF-1α) as well as the activation of kynurenine pathway. Taken together, findings highlight further, the prospects of NPs as alternative source of anti-parasite agents.

Keywords: drug discovery, infectious diseases, mode of action, nanomedicine

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