Search results for: Spread Spectrum Modulation (SSM)

Authors: Elena Ionescu, Elena Iacob, Marie-Louise Ionescu, Carmen Elena Tebrencu, Oana Teodora Ciuperca

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Echinacea species (Asteraceae) has received a global attention because it is widely used for treatment of cold, flu and upper respiratory tract infections. Echinacea species contain a great variety of chemical components that contribute to their activity. The most important components responsible for the biological activity are those with high molecular-weight such as polysaccharides, polyacetylenes, highly unsaturated alkamides and caffeic acid derivatives. The principal factors that may influence the chemical composition of Echinacea include the species and the part of plant used (aerial parts or roots ). In recent years the market for Echinacea has grown rapidly and also the cases of adultery/replacement especially for Echinacea root. The identification of presence or absence of same biomarkers provide information for safe use of Echinacea species in food supplements industry. The aim of the study was the preliminary evaluation and fingerprinting by UV-VISIBLE spectroscopy of biomarkers in terms of content in phenolic derivatives of some Echinacea species (E. purpurea, E. angustifolia and E. pallida) for identification and authentication of the species. The steps of the study were: (1) samples (extracts) preparation from Echinacea species (non-hydrolyzed and hydrolyzed ethanol extracts); (2) samples preparation of reference substances (polyphenol acids: caftaric acid, caffeic acid, chlorogenic acid, ferulic acid; flavonoids: rutoside, hyperoside, isoquercitrin and their aglycones: quercitri, quercetol, luteolin, kaempferol and apigenin); (3) identification of specific absorption at wavelengths between 700-200 nm; (4) identify the phenolic compounds from Echinacea species based on spectral characteristics and the specific absorption; each class of compounds corresponds to a maximum absorption in the UV spectrum. The phytochemical compounds were identified at specific wavelengths between 700-200 nm. The absorption intensities were measured. The obtained results proved that ethanolic extract showed absorption peaks attributed to: phenolic compounds (free phenolic acids and phenolic acids derivatives) registrated between 220-280 nm, unsymmetrical chemical structure compounds (caffeic acid, chlorogenic acid, ferulic acid) with maximum absorption peak and absorption "shoulder" that may be due to substitution of hydroxyl or methoxy group, flavonoid compounds (in free form or glycosides) between 330-360 nm, due to the double bond in position 2,3 and carbonyl group in position 4 flavonols. UV spectra showed two major peaks of absorption (quercetin glycoside, rutin, etc.). The results obtained by UV-VIS spectroscopy has revealed the presence of phenolic derivatives such as cicoric acid (240 nm), caftaric acid (329 nm), caffeic acid (240 nm), rutoside (205 nm), quercetin (255 nm), luteolin (235 nm) in all three species of Echinacea. The echinacoside is absent. This profile mentioned above and the absence of phenolic compound echinacoside leads to the conclusion that species harvested as Echinacea angustifolia and Echinacea pallida are Echinacea purpurea also; It can be said that preliminary fingerprinting of Echinacea species through correspondence with the phenolic derivatives profile can be achieved by UV-VIS spectroscopic investigation, which is an adequate technique for preliminary identification and authentication of Echinacea in medicinal herbs.

Keywords: Echinacea species, Fingerprinting, Phenolic compounds, UV-VIS spectroscopy

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Authors: Bianca Peterson, Tomasz J. Sańko, Carlos C. Bezuidenhout, Johnnie Van Den Berg

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Busseola fusca (Fuller) (Lepidoptera: Noctuidae), the maize stem borer, is a major pest in sub-Saharan Africa. It causes economic damage to maize and sorghum crops and has evolved non-recessive resistance to genetically modified (GM) maize expressing the Cry1Ab insecticidal toxin. Since B. fusca is a non-model organism, very little genomic information is publicly available, and is limited to some cytochrome c oxidase I, cytochrome b, and microsatellite data. The biology of B. fusca is well-described, but still poorly understood. This, in combination with its larval-specific behavior, may pose problems for limiting the spread of current resistant B. fusca populations or preventing resistance evolution in other susceptible populations. As part of on-going research into resistance evolution, B. fusca larvae were collected from Bt and non-Bt maize in South Africa, followed by RNA isolation (15 specimens) and sequencing on the Illumina HiSeq 2500 platform. Quality of reads was assessed with FastQC, after which Trimmomatic was used to trim adapters and remove low quality, short reads. Trinity was used for the de novo assembly, whereas TransRate was used for assembly quality assessment. Transcript identification employed BLAST (BLASTn, BLASTp, and tBLASTx comparisons), for which two libraries (nucleotide and protein) were created from 3.27 million lepidopteran sequences. Several transcripts that have previously been implicated in Cry toxin resistance was identified for B. fusca. These included aminopeptidase N, cadherin, alkaline phosphatase, ATP-binding cassette transporter proteins, and mitogen-activated protein kinase. MEGA7 was used to align these transcripts to reference sequences from Lepidoptera to detect mutations that might potentially be contributing to Cry toxin resistance in this pest. RSEM and Bioconductor were used to perform differential gene expression analysis on groups of B. fusca larvae challenged and unchallenged with the Cry1Ab toxin. Pairwise expression comparisons of transcripts that were at least 16-fold expressed at a false-discovery corrected statistical significance (p) ≤ 0.001 were extracted and visualized in a hierarchically clustered heatmap using R. A total of 329,194 transcripts with an N50 of 1,019 bp were generated from the over 167.5 million high-quality paired-end reads. Furthermore, 110 transcripts were over 10 kbp long, of which the largest one was 29,395 bp. BLAST comparisons resulted in identification of 157,099 (47.72%) transcripts, among which only 3,718 (2.37%) were identified as Cry toxin receptors from lepidopteran insects. According to transcript expression profiles, transcripts were grouped into three subclusters according to the similarity of their expression patterns. Several immune-related transcripts (pathogen recognition receptors, antimicrobial peptides, and inhibitors) were up-regulated in the larvae feeding on Bt maize, indicating an enhanced immune status in response to toxin exposure. Above all, extremely up-regulated arylphorin genes suggest that enhanced epithelial healing is one of the resistance mechanisms employed by B. fusca larvae against the Cry1Ab toxin. This study is the first to provide a resource base and some insights into a potential mechanism of Cry1Ab toxin resistance in B. fusca. Transcriptomic data generated in this study allows identification of genes that can be targeted by biotechnological improvements of GM crops.

Keywords: epithelial healing, Lepidoptera, resistance, transcriptome

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Authors: Syed Jamil H. Kazmi, Faiza Sarwar

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The Indus delta of Sindh Province forms the largest creeks zone of Pakistan. The Sindh coast starts from the mouth of Hab River and terminates at Sir Creek area. In this paper, we have considered the major creeks from the site of Bin Qasim Port in Karachi to Jetty of Keti Bunder in Thatta District. A general decline in the mangrove forest has been observed that within a span of last 25 years. The unprecedented human interventions damage the creeks habitat badly which includes haphazard urban development, industrial and sewage disposal, illegal cutting of mangroves forest, reduced and inconsistent fresh water flow mainly from Jhang and Indus rivers. These activities not only harm the creeks habitat but affected the fish stock substantially. Fishing is the main livelihood of coastal people but with the above-mentioned threats, it is also under enormous pressure by fish catches resulted in unchecked overutilization of the fish resources. This pressure is almost unbearable when it joins with deleterious fishing methods, uncontrolled fleet size, increase trash and by-catch of juvenile and illegal mesh size. Along with these anthropogenic interventions study area is under the red zone of tropical cyclones and active seismicity causing floods, sea intrusion, damage mangroves forests and devastation of fish stock. In order to sustain the natural resources of the Indus Creeks, this study was initiated with the support of FAO, WWF and NIO, the main purpose was to develop a Geo-Spatial dataset for fish stock assessment. The study has been spread over a year (2013-14) on monthly basis which mainly includes detailed fish stock survey, water analysis and few other environmental analyses. Environmental analysis also includes the habitat classification of study area which has done through remote sensing techniques for 22 years’ time series (1992-2014). Furthermore, out of 252 species collected, fifteen species from estuarine and marine groups were short-listed to measure the weight, health and growth of fish species at each creek under GIS data through SPSS system. Furthermore, habitat suitability analysis has been conducted by assessing the surface topographic and aspect derivation through different GIS techniques. The output variables then overlaid in GIS system to measure the creeks productivity. Which provided the results in terms of subsequent classes: extremely productive, highly productive, productive, moderately productive and less productive. This study has revealed the Geospatial tools utilization along with the evaluation of the fisheries resources and creeks habitat risk zone mapping. It has also been identified that the geo-spatial technologies are highly beneficial to identify the areas of high environmental risk in Sindh Creeks. This has been clearly discovered from this study that creeks with high rugosity are more productive than the creeks with low levels of rugosity. The study area has the immense potential to boost the economy of Pakistan in terms of fish export, if geo-spatial techniques are implemented instead of conventional techniques.

Keywords: fish stock, geo-spatial, productivity analysis, risk

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Authors: Pierobon Enrico, Missiaggia Marta, Castelluzzo Michele, Tommasino Francesco, Ricci Leonardo, Scifoni Emanuele, Vincezo Monaco, Boscardin Maurizio, La Tessa Chiara

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Clinical outcomes collected over the past three decades have suggested that ion therapy has the potential to be a treatment modality superior to conventional radiation for several types of cancer, including recurrences, as well as for other diseases. Although the results have been encouraging, numerous treatment uncertainties remain a major obstacle to the full exploitation of particle radiotherapy. To overcome therapy uncertainties optimizing treatment outcome, the best possible radiation quality description is of paramount importance linking radiation physical dose to biological effects. Microdosimetry was developed as a tool to improve the description of radiation quality. By recording the energy deposition at the micrometric scale (the typical size of a cell nucleus), this approach takes into account the non-deterministic nature of atomic and nuclear processes and creates a direct link between the dose deposited by radiation and the biological effect induced. Microdosimeters measure the spectrum of lineal energy y, defined as the energy deposition in the detector divided by most probable track length travelled by radiation. The latter is provided by the so-called “Mean Chord Length” (MCL) approximation, and it is related to the detector geometry. To improve the characterization of the radiation field quality, we define a new quantity replacing the MCL with the actual particle track length inside the microdosimeter. In order to measure this new quantity, we propose a two-stage detector consisting of a commercial Tissue Equivalent Proportional Counter (TEPC) and 4 layers of Low Gain Avalanche Detectors (LGADs) strips. The TEPC detector records the energy deposition in a region equivalent to 2 um of tissue, while the LGADs are very suitable for particle tracking because of the thickness thinnable down to tens of micrometers and fast response to ionizing radiation. The concept of HDM has been investigated and validated with Monte Carlo simulations. Currently, a dedicated readout is under development. This two stages detector will require two different systems to join complementary information for each event: energy deposition in the TEPC and respective track length recorded by LGADs tracker. This challenge is being addressed by implementing SoC (System on Chip) technology, relying on Field Programmable Gated Arrays (FPGAs) based on the Zynq architecture. TEPC readout consists of three different signal amplification legs and is carried out thanks to 3 ADCs mounted on a FPGA board. LGADs activated strip signal is processed thanks to dedicated chips, and finally, the activated strip is stored relying again on FPGA-based solutions. In this work, we will provide a detailed description of HDM geometry and the SoC solutions that we are implementing for the readout.

Keywords: particle tracking, ion therapy, low gain avalanche diode, tissue equivalent proportional counter, microdosimetry

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Authors: Chunyang Miao, Bingxin Li, Shanglong Ning, Christopher J. B. Ford

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While it is challenging to fabricate electronic devices close to atomic dimensions in conventional top-down lithography, molecular electronics is promising to help maintain the exponential increase in component densities via using molecular building blocks to fabricate electronic components from the bottom up. It offers smaller, faster, and more energy-efficient electronic and photonic systems. A self-assembled monolayer (SAM) of molecules is a layer of molecules that self-assembles on a substrate. They are mechanically flexible, optically transparent, low-cost, and easy to fabricate. A large-area multi-layer structure has been designed and investigated by the team, where a SAM of designed molecules is sandwiched between graphene and gold electrodes. Each molecule can act as a quantum dot, with all molecules conducting in parallel. When a source-drain bias is applied, significant current flows only if a molecular orbital (HOMO or LUMO) lies within the source-drain energy window. If electrons tunnel sequentially on and off the molecule, the charge on the molecule is well-defined and the finite charging energy causes Coulomb blockade of transport until the molecular orbital comes within the energy window. This produces ‘Coulomb diamonds’ in the conductance vs source-drain and gate voltages. For different tunnel barriers at either end of the molecule, it is harder for electrons to tunnel out of the dot than in (or vice versa), resulting in the accumulation of two or more charges and a ‘Coulomb staircase’ in the current vs voltage. This nanostructure exhibits highly reproducible Coulomb-staircase patterns, together with additional oscillations, which are believed to be attributed to molecular vibrations. Molecules are more isolated than semiconductor dots, and so have a discrete phonon spectrum. When tunnelling into or out of a molecule, one or more vibronic states can be excited in the molecule, providing additional transport channels and resulting in additional peaks in the conductance. For useful molecular electronic devices, achieving the optimum orbital alignment of molecules to the Fermi energy in the leads is essential. To explore it, a drop of ionic liquid is employed on top of the graphene to establish an electric field at the graphene, which screens poorly, gating the molecules underneath. Results for various molecules with different alignments of Fermi energy to HOMO have shown highly reproducible Coulomb-diamond patterns, which agree reasonably with DFT calculations. In summary, this large-area SAM molecular junction is a promising candidate for future electronic circuits. (1) The small size (1-10nm) of the molecules and good flexibility of the SAM lead to the scalable assembly of ultra-high densities of functional molecules, with advantages in cost, efficiency, and power dissipation. (2) The contacting technique using graphene enables mass fabrication. (3) Its well-observed Coulomb blockade behaviour, narrow molecular resonances, and well-resolved vibronic states offer good tuneability for various functionalities, such as switches, thermoelectric generators, and memristors, etc.

Keywords: molecular electronics, Coulomb blokade, electron-phonon coupling, self-assembled monolayer

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Authors: Irene Carmagnola, Valeria Chiono, Sandra Pacharra, Jochen Salber, Sean McMahon, Chris Lovell, Pooja Basnett, Barbara Lukasiewicz, Ipsita Roy, Xiang Zhang, Gianluca Ciardelli

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Thrombosis and restenosis after stenting procedure can be prevented by promoting fast stent wall endothelialisation. It is well known that surface functionalisation with antifouling molecules combining with extracellular matrix proteins is a promising strategy to design biomimetic surfaces able to promote fast endothelialization. In particular, REDV has gained much attention for the ability to enhance rapid endothelialization due to its specific affinity with endothelial cells (ECs). In this work, a two-step plasma treatment was performed to polymerize a thin layer of acrylic acid, used to subsequently graft PEGylated-REDV and polyethylene glycol (PEG) at different molar ratio with the aim to selectively promote endothelial cell adhesion avoiding platelet activation. PEGylate-REDV was provided by Biomatik and it is formed by 6 PEG monomer repetitions (Chempep Inc.), with an NH2 terminal group. PEG polymers were purchased from Chempep Inc. with two different chain lengths: m-PEG6-NH2 (295.4 Da) with 6 monomer repetitions and m-PEG12-NH2 (559.7 Da) with 12 monomer repetitions. Plasma activation was obtained by operating at 50W power, 5 min of treatment and at an Ar flow rate of 20 sccm. Pure acrylic acid (99%, AAc) vapors were diluted in Ar (flow = 20 sccm) and polymerized by a pulsed plasma discharge applying a discharge RF power of 200 W, a duty cycle of 10% (on time = 10 ms, off time = 90 ms) for 10 min. After plasma treatment, samples were dipped into an 1-(3-dimethylaminopropyl)-3- ethylcarbodiimide (EDC)/N-hydroxysuccinimide (NHS) solution (ratio 4:1, pH 5.5) for 1 h at 4°C and subsequently dipped in PEGylate-REDV and PEGylate-REDV:PEG solutions at different molar ratio (100 μg/mL in PBS) for 20 h at room temperature. Surface modification was characterized through physico-chemical analyses and in vitro cell tests. PEGylated-REDV peptide and PEG were successfully bound to the carboxylic groups that are formed on the polymer surface after plasma reaction. FTIR-ATR spectroscopy, X -ray Photoelectron Spectroscopy (XPS) and contact angle measurement gave a clear indication of the presence of the grafted molecules. The use of PEG as a spacer allowed for an increase in wettability of the surface, and the effect was more evident by increasing the amount of PEG. Endothelial cells adhered and spread well on the surfaces functionalized with the REDV sequence. In conclusion, a selective coating able to promote a new endothelial cell layer on polymeric stent surface was developed. In particular, a thin AAc film was polymerised on the polymeric surface in order to expose –COOH groups, and PEGylate-REDV and PEG were successful grafted on the polymeric substrates. The REDV peptide demonstrated to encourage cell adhesion with a consequent, expected improvement of the hemocompatibility of these polymeric surfaces in vivo. Acknowledgements— This work was funded by the European Commission 7th Framework Programme under grant agreement number 604251- ReBioStent (Reinforced Bioresorbable Biomaterials for Therapeutic Drug Eluting Stents). The authors thank all the ReBioStent partners for their support in this work.

Keywords: endothelialisation, plasma treatment, stent, surface functionalisation

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Authors: Saeid H. Mobini, Monika Lulsdorf, Thomas D. Warkentin

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The length of the breeding cycle from seed to seed is a limiting factor in the development of improved homozygous lines for breeding or recombinant inbred lines (RILs) for genetic analysis. The objective of this research was to accelerate the production of field pea RILs through application of rapid generation technology (RGT). RGT is based on the principle of growing miniature plants in an artificial medium under controlled conditions, and allowing them to produce a few flowers which develop seeds that are harvested prior to normal seed maturity. We aimed to maintain population size and genetic diversity in regeneration cycles. The effects of flurprimidol (a gibberellin synthesis inhibitor), plant density, hydroponic system, scheduled fertilizer applications, artificial light spectrum, photoperiod, and light/dark temperature were evaluated in the development of RILs from a cross between cultivars CDC Dakota and CDC Amarillo. The main goal was to accelerate flowering while reducing maintenance and space costs. In addition, embryo rescue of immature seeds was tested for shortening the seed fill period. Data collected over seven generations included plant height, the percentage of plant survival, flowering rate, seed setting rate, the number of seeds per plant, and time from seed to seed. Applying 0.6 µM flurprimidol reduced the internode length. Plant height was decreased to approximately 32 cm allowing for higher plant density without a delay in flowering and seed setting rate. The three light systems (T5 fluorescent bulbs, LEDs, and High Pressure Sodium +Metal-halide lamp) evaluated did not differ significantly in terms of flowering time in field pea. Collectively, the combination of 0.6 µM flurprimidol, 217 plant. m-2, 20 h photoperiod, 21/16 oC light/dark temperature in a hydroponic system with vermiculite substrate, applying scheduled fertilizer application based on growth stage, and 500 µmole.m-2.s-1 light intensity using T5 bulbs resulted in 100% of plants flowering within 34 ± 3 days and 96.5% of plants completed seed setting in 68.2 ± 3.6 days, i.e., 30-45 days/generation faster than conventional single seed descent (SSD) methods. These regeneration cycles were reproducible consistently. Hence, RGT could double (5.3) generations per year, using 3% occupying space, compared to SSD (2-3 generation/year). Embryo rescue of immature seeds at 7-8 mm stage, using commercial fertilizer solutions (Holland’s Secret™) showed seed setting rate of 95%, while younger embryos had lower germination rate. Mature embryos had a seed setting rate of 96.5% without either hormones or sugar added. So, considering the higher cost of embryo rescue using a procedure which requires skill, additional materials, and expenses, it could be removed from RGT with a further cost saving, and the process could be stopped between generations if required.

Keywords: field pea, flowering, rapid regeneration, recombinant inbred lines, single seed descent

Procedia PDF Downloads 342

Authors: Maryam Hajrezaie, Mahmood Ameen Abdulla, Nazia Abdul Majid, Hapipa Mohd Ali, Pouya Hassandarvish, Maryam Zahedi Fard

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Background: Colon cancer is one of the most prevalent cancers in the world and is the third leading cause of death among cancers in both males and females. The incidence of colon cancer is ranked fourth among all cancers but varies in different parts of the world. Cancer chemoprevention is defined as the use of natural or synthetic compounds capable of inducing biological mechanisms necessary to preserve genomic fidelity. Andrographolide is the major labdane diterpenoidal constituent of the plant Andrographis paniculata (family Acanthaceae), used extensively in the traditional medicine. Extracts of the plant and their constituents are reported to exhibit a wide spectrum of biological activities of therapeutic importance. Laboratory animal model studies have provided evidence that Andrographolide play a role in inhibiting the risk of certain cancers. Objective: Our aim was to evaluate the chemopreventive efficacy of the Andrographolide in the AOM induced rat model. Methods: To evaluate inhibitory properties of andrographolide on colonic aberrant crypt foci (ACF), five groups of 7-week-old male rats were used. Group 1 (control group) were fed with 10% Tween 20 once a day, Group 2 (cancer control) rats were intra-peritoneally injected with 15 mg/kg Azoxymethan, Gropu 3 (drug control) rats were injected with 15 mg/kg azoxymethan and 5-Flourouracil, Group 4 and 5 (experimental groups) were fed with 10 and 20 mg/kg andrographolide each once a day. After 1 week, the treatment group rats received subcutaneous injections of azoxymethane, 15 mg/kg body weight, once weekly for 2 weeks. Control rats were continued on Tween 20 feeding once a day and experimental groups 10 and 20 mg/kg andrographolide feeding once a day for 8 weeks. All rats were sacrificed 8 weeks after the azoxymethane treatment. Colons were evaluated grossly and histopathologically for ACF. Results: Administration of 10 mg/kg and 20 mg/kg andrographolide were found to be effectively chemoprotective, as evidenced microscopily and biochemically. Andrographolide suppressed total colonic ACF formation up to 40% to 60%, respectively, when compared with control group. Pre-treatment with andrographolide, significantly reduced the impact of AOM toxicity on plasma protein and urea levels as well as on plasma aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH) and gamma-glutamyl transpeptidase (GGT) activities. Grossly, colorectal specimens revealed that andrographolide treatments decreased the mean score of number of crypts in AOM-treated rats. Importantly, rats fed andrographolide showed 75% inhibition of foci containing four or more aberrant crypts. The results also showed a significant increase in glutathione (GSH), superoxide dismutase (SOD), nitric oxide (NO), and Prostaglandin E2 (PGE2) activities and a decrease in malondialdehyde (MDA) level. Histologically all treatment groups showed a significant decrease of dysplasia as compared to control group. Immunohistochemical staining showed up-regulation of Hsp70 and down-regulation of Bax proteins. Conclusion: The current study demonstrated that Andrographolide reduce the number of ACF. According to these data, Andrographolide might be a promising chemoprotective activity, in a model of AOM-induced in ACF.

Keywords: chemopreventive, andrographolide, colon cancer, aberrant crypt foci (ACF)

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Authors: Tramuta Clara, Masotti Chiara, Pitti Monica, Adriano Daniela, Battistini Roberta, Serraca Laura, Decastelli Lucia

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Oysters are considered filter organisms, and their raw consumption may increase health risks for consumers: it is often associated with outbreaks of gastroenteritis or enteric illnesses. Most of these foodborne diseases are caused by Vibrio strains, enteric pathogens also involved in the diffusion of genetic determinants of antibiotic resistance and their entrance along the food chain. The European Food Safety Authority (EFSA), during the European Union report on antimicrobial resistance in 2017, focused the attention about the role of food as a possible carrier of antibiotic-resistant bacteria or antibiotic-resistance genes that determine health risks for humans. This study wants to determine antibiotic resistance and antibiotic-resistance genes in Vibrio spp. isolated from Crassostrea gigas oysters collected in the Golfo della Spezia (Liguria, Italy). A total of 47 Vibrio spp. strains were isolated (ISO21872-2:2017) during the summer of 2021 from oysters of Crassostrea gigas. The strains were identified by MALDI-TOF (Bruker, Germany) mass spectrometry and tested for antibiotic susceptibility using a broth microdiluition method (ISO20776-1:2019) using Sensititre EUVSEC plates (Thermo-Fisher Scientific) to obtain the Minimum Inhibitory Concentration (MIC). The strains were tested with PCR-based biomolecular methods, according to previous works, to define the presence of 23 resistance genes of the main classes of antibiotics used in human and veterinary medicine: tet (B), tet (C), tet (D), tet (A), tet (E), tet (G ), tet (K), tet (L), tet (M), tet (O), tet (S) (tetracycline resistance); blaCTX-M, blaTEM, blaOXA, blaSHV (β-lactam resistance); mcr-1 and mcr-2 (colistin resistance); qnrA, qnrB, and qnrS (quinolone resistance); sul1, sul2 and sul3 (sulfonamide resistance). Six different species have been identified: V. alginolyticus 34% (n=16), V. harveyi 28% (n=13), V. fortis 15% (n=7), V. pelagius 8% (n=4), V. parahaemolyticus 11% (n=5) e V. chagasii 4% (n=2). The PCR assays showed the presence of the blaTEM gene on 40% of the strains (n=19). All the other genes were not detected, except for a V. alginolyticus positive for anrS gene. The broth microdiluition method results showed an high level of resistance for ciprofloxacin (62%; n=29), ampicillin (47%; n=22), and colistin (49%; n=23). Furthermore, 32% (n=15) of strains can be considered multiresistant bacteria for the simultaneous presence of resistance for three different antibiotic classes. Susceptibility towards meropenem, azithromycin, gentamicin, ceftazidime, cefotaxime, chloramphenicol, tetracycline and sulphamethoxazole reached 100%. The Vibrio species identified in this study are widespread in marine environments and can cause gastrointerstinal infections after the ingestion of raw fish products and bivalve molluscs. The level of resistance to antibiotics such as ampicillin, ciprofloxacin and colistin can be connected to anthropic factors (industrial, agricultural and domestic wastes) that promote the spread of resistance to these antibiotics. It can be also observed a strong correlation between phenotypic (resistant MIC) and genotypic (positive blaTEM gene) resistance for ampicillin on the same strains, probably due to the transfer of genetic material between bacterial strains. Consumption of raw bivalve molluscs can represent a risk for consumers heath due to the potentially presence of foodborne pathogens, highly resistant to different antibiotics and source of transferable antibiotic-resistant genes.

Keywords: vibrio species, blaTEM genes, antimicrobial resistance, PCR

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Authors: Sana Fatima, Paul Sadler, Jon Cooper, David Mendel, Ayesha Jameel

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Background: Neurodiversity refers to intrinsic differences between human minds and encompasses dyspraxia, dyslexia, attention deficit hyperactivity disorder, dyscalculia, autism spectrum disorder, and Tourette syndrome. There is increasing recognition of neurodiversity in relation to disability/diversity in medical education and the associated impact on training, career progression, and personal and professional wellbeing. In addition, documented and anecdotal evidence suggests that medical educators and training providers in all four nations (UK) are increasingly concerned about understanding neurodiversity and identifying and providing support for neurodivergent trainees. Summary of Work: A national Neurodiversity Task and Finish group were established to survey Health Education England local office Professional Support teams about insights into infrastructure, training for educators, triggers for assessment, resources, and intervention protocols. This group drew from educational leadership, professional and personal neurodiverse expertise, occupational medicine, employer human resource, and trainees. An online, exploratory survey was conducted to gather insights from supervisors and trainers across England using the Professional Support Units' platform. Summary of Results: This survey highlighted marked heterogeneity in the identification, assessment, and approaches to support and management of neurodivergent trainees and highlighted a 'deficit' approach to neurodiversity. It also demonstrated a paucity of educational and protocol resources for educators and supervisors in supporting neurodivergent trainees. Discussions and Conclusions: In phase one, we focused on faculty development. An educational repository for all supervising trainees using a thematic approach was formalised. This was guided by our survey findings specific for neurodiversity and took a triple 'A' approach: awareness, assessment, and action. This is further supported by video material incorporating stories in training as well as mobile workshops for trainers for more immersive learning. The subtle theme from both the survey and Task and finish group suggested a move away from deficit-focused methods toward a positive holistic, interdisciplinary approach within a biopsychosocial framework. Contributions: 1. Faculty Knowledge and basic understanding of neurodiversity are key to supporting trainees with known or underlying Neurodiverse conditions. This is further complicated by challenges around non-disclosure, varied presentations, stigma, and intersectionality. 2. There is national (and international) inconsistency in the approach to how trainees are managed once a neurodiverse condition is suspected or diagnosed. 3. A carefully constituted and focussed Task and Finish group can rapidly identify national inconsistencies in neurodiversity and implement rapid educational interventions. 4. Nuanced findings from surveys and discussion can reframe the approach to neurodiversity; from a medical model to a more comprehensive, asset-based, biopsychosocial model of support, fostering a cultural shift, accepting 'diversity' in all its manifestations, visible and hidden.

Keywords: neurodiversity, professional support, human considerations, workplace wellbeing

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Authors: Deepak Loura

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Production and productivity of several crops in the country continue to be adversely affected by biotic (e.g., Insect-pests and diseases) and abiotic (e.g., water temperature and salinity) stresses. Over-dependence on pesticides and other chemicals is economically non-viable for the resource-poor farmers of our country. Further, pesticides can potentially affect human and environmental safety. While traditional breeding techniques and proper- management strategies continue to play a vital role in crop improvement, we need to judiciously use biotechnology approaches for the development of genetically modified crops addressing critical problems in the improvement of crop plants for sustainable agriculture. Modern biotechnology can help to increase crop production, reduce farming costs, and improve food quality and the safety of the environment. Genetic engineering is a new technology which allows plant breeders to produce plants with new gene combinations by genetic transformation of crop plants for improvement of agronomic traits. Advances in recombinant DNA technology have made it possible to have genes between widely divergent species to develop genetically modified or genetically engineered plants. Plant genetic engineering provides the strength to harness useful genes and alleles from indigenous microorganisms to enrich the gene pool for developing genetically modified (GM) crops that will have inbuilt (inherent) resistance to insect pests, diseases, and abiotic stresses. Plant biotechnology has made significant contributions in the past 20 years in the development of genetically engineered or genetically modified crops with multiple benefits. A variety of traits have been introduced in genetically engineered crops which include (i) herbicide resistance. (ii) pest resistance, (iii) viral resistance, (iv) slow ripening of fruits and vegetables, (v) fungal and bacterial resistance, (vi) abiotic stress tolerance (drought, salinity, temperature, flooding, etc.). (vii) quality improvement (starch, protein, and oil), (viii) value addition (vitamins, micro, and macro elements), (ix) pharmaceutical and therapeutic proteins, and (x) edible vaccines, etc. Multiple genes in transgenic crops can be useful in developing durable disease resistance and a broad insect-control spectrum and could lead to potential cost-saving advantages for farmers. The development of transgenic to produce high-value pharmaceuticals and the edible vaccine is also under progress, which requires much more research and development work before commercially viable products will be available. In addition, molecular-aided selection (MAS) is now routinely used to enhance the speed and precision of plant breeding. Newer technologies need to be developed and deployed for enhancing and sustaining agricultural productivity. There is a need to optimize the use of biotechnology in conjunction with conventional technologies to achieve higher productivity with fewer resources. Therefore, genetic modification/ engineering of crop plants assumes greater importance, which demands the development and adoption of newer technology for the genetic improvement of crops for increasing crop productivity.

Keywords: biotechnology, plant genetic engineering, genetically modified, biotic, abiotic, disease resistance

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Authors: Devin Doos, Ashley Hughes, Trang Pham, Paul Barach, Rami Ahmed

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Over 115,000 Health Care Workers (HCWs) have died from COVID-19, and millions have been infected while caring for patients. HCWs have filed thousands of safety complaints surrounding safety concerns due to Personal Protective Equipment (PPE) shortages, which included concerns around inadequate and PPE reuse. Protocols for donning and doffing PPE remain ambiguous, lacking an evidence-base, and often result in wide deviations in practice. PPE donning and doffing protocol deviations commonly result in self-contamination but have not been thoroughly addressed. No evidence-driven protocols provide guidance on protecting HCW during periods of PPE reuse. Objective: The aim of this study was to examine safety-related threats and risks to Health Care Workers (HCWs) due to the reuse of PPE among Emergency Department personnel. Method: We conducted a prospective observational study to examine the risks of reusing PPE. First, ED personnel were asked to don and doff PPE in a simulation lab. Each participant was asked to don and doff PPE five times, according to the maximum reuse recommendation set by the Centers for Disease Control and Prevention (CDC). Each participant was videorecorded; video recordings were reviewed and coded independently by at least 2 of the 3trained coders for safety behaviors and riskiness of actions. A third coder was brought in when the agreement between the 2 coders could not be reached. Agreement between coders was high (81.9%), and all disagreements (100%) were resolved via consensus. A bowtie risk assessment chart was constructed analyzing the factors that contribute to increased risks HCW are faced with due to PPE use and reuse. Agreement amongst content experts in the field of Emergency Medicine, Human Factors, and Anesthesiology was used to select aspects of health care that both contribute and mitigate risks associated with PPE reuse. Findings: Twenty-eight clinician participants completed five rounds of donning/doffing PPE, yielding 140 PPE donning/doffing sequences. Two emerging threats were associated with behaviors in donning, doffing, and re-using PPE: (i) direct exposure to contaminant, and (ii) transmission/spread of contaminant. Protective behaviors included: hand hygiene, not touching the patient-facing surface of PPE, and ensuring a proper fit and closure of all PPE materials. 100% of participants (n= 28) deviated from the CDC recommended order, and most participants (92.85%, n=26) self-contaminated at least once during reuse. Other frequent errors included failure to tie all ties on the PPE (92.85%, n=26) and failure to wash hands after a contamination event occurred (39.28%, n=11). Conclusions: There is wide variation and regular errors in how HCW don and doffPPE while including in reusing PPE that led to self-contamination. Some errors were deemed “recoverable”, such as hand washing after touching a patient-facing surface to remove the contaminant. Other errors, such as using a contaminated mask and accidentally spreading to the neck and face, can lead to compound risks that are unique to repeated PPE use. A more comprehensive understanding of the contributing threats to HCW safety and complete approach to mitigating underlying risks, including visualizing with risk management toolsmay, aid future PPE designand workflow and space solutions.

Keywords: bowtie analysis, health care, PPE reuse, risk management

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Authors: Nils Haglund, Svenolof Dahlgren, Maria Rastam, Peik Gustafsson, Karin Kalien

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In Sweden, like in most developed countries, there is a substantial increase of children diagnosed with autism and other conditions within the autism spectrum (ASD). The rapid increase of ASD rates stresses the importance of developing care programs to provide support and comprehensive interventions for affected families. The current observational study was conducted in order to evaluate an ongoing Comprehensive Intensive Early Intervention (CIEI) program for children with autism in southern Sweden. The change in autism symptoms among children participating in CIEI (intervention group, n=67) was compared with children who received traditional habilitation services only (comparison group, n=27). Children of parents who accepted the offered CIEI-program, constituted the intervention group, whereas children, whose parents (for some reason) were not interested in the offered CIEI-program, constituted the comparison group. The CIEI-program was individualized to each child by experienced applied behavior analysis (ABA) specialists with different backgrounds as psychologists, speech pathologists or special education teachers, in cooperation with parents and preschool staff. Due to the individualization, the intervention could vary in intensity and techniques. The intensity was calculated to 15-25 hours each week at home and the preschool altogether. Each child was assigned one 'trainer', who was often employed as a preschool teacher but could have another educational background. An agreement between supervisor- parents and preschool staff was reached to confirm the intensity and content of the CIEI- program over an approximately two-year intervention period. Symptom changes were measured as evaluation-ADOS-2-scores, total- and severity-scores, minus the corresponding baseline-scores, divided by the time between baseline and evaluation. The difference between the study-groups regarding change of ADOS-2-scores was estimated using ANCOVA. In the current study, children in the CIEI-group improved their ADOS-2-total scores between baseline and evaluation (-0.8 scores per year; 95%CI: -1.2 to -0.4), whereas no such improvement was detected in the comparison group (+0.1 scores per year; 95%CI: -0.7 to +0.9). The change difference (change in the CIEI-group vs. change in the comparison group) was statistically significant, both crude and after adjusting for possible confounders (-1.1; 95%CI -1.9 to -0.4). Children in the CIEI-group also significantly improved their ADOS-calibrated severity scores, but not significantly differently so from the comparison group. The results from the current study indicate that the CIEI program significantly improves social and communicative skills among children with autism and that children with developmental delay could benefit to a similar degree as other children. The results support earlier studies reporting on the improvement of autism symptoms after early intensive interventions. The results from observational studies are difficult to interpret, but it is nevertheless of uttermost importance to evaluate costly autism intervention programs. Such results may be of immediate importance to healthcare organizations when allocating the already strained resources to different patient groups. Albeit the obvious limitation of the current naturalistic study, the results support previous positive studies and indicate that children with autism benefit from participating in early comprehensive, intensive programs and that investments in these programs may be highly justifiable.

Keywords: autism symptoms, ADOS-scores, evaluation, intervention program

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Authors: Rupesh S. Gundewar, Kanchan C. Khare

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Runoff contribution from urban areas is generally from manmade structures and few natural contributors. The manmade structures are buildings; roads and other paved areas whereas natural contributors are groundwater and overland flows etc. Runoff alleviation is done by manmade as well as natural storages. Manmade storages are storage tanks or other storage structures such as soakways or soak pits which are more common in western and European countries. Natural storages are catchment slope, infiltration, catchment length, channel rerouting, drainage density, depression storage etc. A literature survey on the manmade and natural storages/inflow has presented percentage contribution of each individually. Sanders et.al. in their research have reported that a vegetation canopy reduces runoff by 7% to 12%. Nassif et el in their research have reported that catchment slope has an impact of 16% on bare standard soil and 24% on grassed soil on rainfall runoff. Infiltration being a pervious/impervious ratio dependent parameter is catchment specific. But a literature survey has presented a range of 15% to 30% loss of rainfall runoff in various catchment study areas. Catchment length and channel rerouting too play a considerable role in reduction of rainfall runoff. Ground infiltration inflow adds to the runoff where the groundwater table is very shallow and soil saturates even in a lower intensity storm. An approximate percent contribution through this inflow and surface inflow contributes to about 2% of total runoff volume. Considering the various contributing factors in runoff it has been observed during a literature survey that integrated modelling approach needs to be considered. The traditional storm water network models are able to predict to a fair/acceptable degree of accuracy provided no interaction with receiving water (river, sea, canal etc), ground infiltration, treatment works etc. are assumed. When such interactions are significant then it becomes difficult to reproduce the actual flood extent using the traditional discrete modelling approach. As a result the correct flooding situation is very rarely addressed accurately. Since the development of spatially distributed hydrologic model the predictions have become more accurate at the cost of requiring more accurate spatial information.The integrated approach provides a greater understanding of performance of the entire catchment. It enables to identify the source of flow in the system, understand how it is conveyed and also its impact on the receiving body. It also confirms important pain points, hydraulic controls and the source of flooding which could not be easily understood with discrete modelling approach. This also enables the decision makers to identify solutions which can be spread throughout the catchment rather than being concentrated at single point where the problem exists. Thus it can be concluded from the literature survey that the representation of urban details can be a key differentiator to the successful understanding of flooding issue. The intent of this study is to accurately predict the runoff from impermeable areas from urban area in India. A representative area has been selected for which data was available and predictions have been made which are corroborated with the actual measured data.

Keywords: runoff, urbanization, impermeable response, flooding

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Authors: Ivana Cabarkapa, Zorica Tomicic, Olivera Duragic

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Antimicrobial resistance represents one of the major challenges facing humanity in the last decades. Increasing antibiotic-resistant pathogens indicates the need for the development of alternative antibacterial drugs and new treatment strategies. One of the innovative emerging treatments in overcoming multidrug-resistant pathogens certainly represents the inhibition anti-quorum sensing system. For most of the food-borne pathogens, the expression of the virulence depends on their capability communication with other members of the population by means of quorum sensing (QS). QS represents a specific way of bacterial intercellular communication, which enabled owing to their ability to detect and to respond to cell population density by gene regulation. QS mechanisms are responsible for controls the pathogenesis, virulence luminescence, motility, sporulation and biofilm formation of many organisms by regulating gene expression. Therefore, research in this field is being an attractive target for the development of new natural antibacterial agents. Anti-QS compounds are known to have the ability to prohibit bacterial pathogenicity. Considering the importance of quorum sensing during bacterial pathogenesis, this research has been focused on evaluation anti - QS properties of four essential oils (EOs) Origanum heracleoticum, Origanum vulgare, Thymus vulgare, and Thymus serpyllum, using biomonitor strain of Chromobacterium violaceum CV026. Tests conducted on Luria Bertani agar supplemented with N hexanol DL homoserine lacton (HHL) 10µl/50ml of agar. The anti-QS potential of the EOs was assayed in a range of concentrations of 200 – 0.39 µl/ml using the disc diffusion method. EOs of Th. vulgaris and T. serpyllum were exhibited anti-QS activity indicated by a non- pigmented ring with a dilution-dependent manner. The lowest dilution of EOs T. vulgaris and T. serpyllum in which they exhibited visually detectable inhibition of violacein synthesis was 6.25 µl/ml for both tested EOs. EOs of O. heracleoticum and O. vulgare were displayed different active principles, i.e., antimicrobial activity indicated by the inner clear ring and anti-QS activity indicated by the outer non-pigmented ring, in a concentration-dependent manner. The lowest dilution of EOs of O. heracleoticum and O. vulgare in which exhibited visually detectable inhibition of violacein synthesis was 1.56 and 3.25 µl/ml, respectively. Considering that, the main constituents of the tested EOs represented by monoterpenes (carvacrol, thymol, γ-terpinene, and p-cymene), anti - QS properties of tested EOs can be mainly attributed to their activity. In particular, from the scientific literature, carvacrol and thymol show a sub-inhibitory effect against foodborne pathogens. Previous studies indicated that sub-lethal concentrations of carvacrol reduced the mobility of bacteria due to the ability of interference using QS mechanism between the bacterial cells, and thereby reducing the ability of biofilm formation The precise mechanism by which carvacrol inhibits biofilm formation is still not fully understood. Our results indicated that EOs displayed different active principles, i.e., antimicrobial activity indicated by the inner clear ring and anti-QS activity indicated by an outer non- pigmented ring with visually detectable inhibition of violacein. Preliminary results suggest that EOs represent a promising alternative for effective control of the emergence and spread of resistant pathogens.

Keywords: anti-quorum sensing activity, Chromobacterium violaceum, essential oils, violacein

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Authors: Savita Tangwongkit, Chittur S Srinivasan, Philip J. Jones

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Thailand has become the world’s fourth largest sugarcane producer and second largest sugar exporter. While there have been a number of drivers of this growth, the primary driver has been wide-ranging government support measures. Recently, the Thai government has emphasized the need for policy reform as part of a broader industry restructuring to bring the sector up-to-date with the current and future developments in the international sugar market. Because of the sectors historical dependence on government support, any such reform is likely to have a very significant impact on the fortunes of Thai cane farmers. This study explores the impact of three policy scenarios, representing a spectrum of policy approaches, on Thai cane producers. These reform scenarios were designed in consultation with policy makers and academics working in the cane sector. Scenario 1 captures the current ‘government proposal’ for policy reform. This scenario removes certain domestic production subsidies but seeks to maintain as much support as is permissible under current WTO rules. The second scenario, ‘protectionism’, maintains the current internal market producer supports, but otherwise complies with international (WTO) commitments. Third, the ‘libertarian scenario’ removes all production support and market interventions, trade and domestic consumption distortions. Most important driver of producer behaviour in all of the scenarios is the producer price of cane. Cane price is obviously highest under the protectionism scenario, followed by government proposal and libertarian scenarios, respectively. Likely producer responses to these three policy scenarios was determined by means of a large-scale survey of cane farmers. The sample was stratified by size group and quotas filled by size group and region. One scenario was presented to each of three sub-samples, consisting of approx.150 farmers. Total sample size was 462 farms. Data was collected by face-to-face interview between June and August 2019. There was a marked difference in farmer response to the three scenarios. Farmers in the ‘Protectionism’ scenario, which maintains the highest cane price and those who farm larger cane areas are more likely to continue cane farming. The libertarian scenario is likely to result in the greatest losses in terms of cane production volume broadly double that of the ‘protectionism’ scenario, primarily due to farmers quitting cane production altogether. Over half of loss cane production volume comes from medium-size farm, i.e. the largest and smallest producers are the most resilient. This result is likely due to the fact that the medium size group are large enough to require hired labour but lack the economies of scale of the largest farms. Over all size groups the farms most heavily specialized in cane production, i.e. those devoting 26-50% of arable land to cane, are also the most vulnerable, with 70% of all farmers quitting cane production coming from this group. This investigation suggests that cane price is the most significant determinant of farmer behaviour. Also, that where scenarios drive significantly lower cane price, policy makers should target support towards mid-sized producers, with policies that encourage efficiency gains and diversification into alternative agricultural crops.

Keywords: farmer intentions, farm survey, policy reform, Thai cane production

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Authors: Richard McGregor, Carmen Reaiche, Stephen Boyle

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Cyber threats are a relatively recent phenomenon and offer cyber insurers a dynamic and intelligent peril. As individuals en mass become increasingly digitally dependent, Personal Cyber Insurance (PCI) offers an attractive option to mitigate cyber risk at a personal level. This abstract proposes a literature review that conceptualises a framework for siting Personal Cyber Insurance (PCI) within the context of cyberspace. The lack of empirical research within this domain demonstrates an immediate need to define the scope of PCI to allow cyber insurers to understand personal cyber risk threats and vectors, customer awareness, capabilities, and their associated needs. Additionally, this will allow cyber insurers to conceptualise appropriate frameworks allowing effective management and distribution of PCI products and services within a landscape often in-congruent with risk attributes commonly associated with traditional personal line insurance products. Cyberspace has provided significant improvement to the quality of social connectivity and productivity during past decades and allowed enormous capability uplift of information sharing and communication between people and communities. Conversely, personal digital dependency furnish ample opportunities for adverse cyber events such as data breaches and cyber-attacksthus introducing a continuous and insidious threat of omnipresent cyber risk–particularly since the advent of the COVID-19 pandemic and wide-spread adoption of ‘work-from-home’ practices. Recognition of escalating inter-dependencies, vulnerabilities and inadequate personal cyber behaviours have prompted efforts by businesses and individuals alike to investigate strategies and tactics to mitigate cyber risk – of which cyber insurance is a viable, cost-effective option. It is argued that, ceteris parabus, the nature of cyberspace intrinsically provides characteristic peculiarities that pose significant and bespoke challenges to cyber insurers, often in-congruent with risk attributes commonly associated with traditional personal line insurance products. These challenges include (inter alia) a paucity of historical claim/loss data for underwriting and pricing purposes, interdependencies of cyber architecture promoting high correlation of cyber risk, difficulties in evaluating cyber risk, intangibility of risk assets (such as data, reputation), lack of standardisation across the industry, high and undetermined tail risks, and moral hazard among others. This study proposes a thematic overview of the literature deemed necessary to conceptualise the challenges to issuing personal cyber coverage. There is an evident absence of empirical research appertaining to PCI and the design of operational business models for this business domain, especially qualitative initiatives that (1) attempt to define the scope of the peril, (2) secure an understanding of the needs of both cyber insurer and customer, and (3) to identify elements pivotal to effective management and profitable distribution of PCI - leading to an argument proposed by the author that postulates that the traditional general insurance customer journey and business model are ill-suited for the lineaments of cyberspace. The findings of the review confirm significant gaps in contemporary research within the domain of personal cyber insurance.

Keywords: cyberspace, personal cyber risk, personal cyber insurance, customer journey, business model

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Authors: Mohamad F. Jamiluddin, Emeline Sarry, Ana Bejanariu, Cécile Bauche

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Introduction: Over 360 million people are chronically infected with hepatitis B virus (HBV), of whom 1 million die each year from HBV-associated liver cirrhosis or hepatocellular carcinoma. Current treatment options for chronic hepatitis B depend on interferon-α (IFNα) or nucleos(t)ide analogs, which control virus replication but rarely eliminate the virus. Treatment with PEG-IFNα leads to a sustained antiviral response in only one third of patients. After withdrawal of the drugs, the rebound of viremia is observed in the majority of patients. Furthermore, the long-term treatment is subsequently associated with the appearance of drug resistant HBV strains that is often the cause of the therapy failure. Among the new therapeutic avenues being developed, therapeutic vaccine aimed at inducing immune responses similar to those found in resolvers is of growing interest. The high prevalence of chronic hepatitis B necessitates the design of better vaccination strategies capable of eliciting broad-spectrum of cell-mediated immunity(CMI) and humoral immune response that can control chronic hepatitis B. Induction of HBV-specific T cells and B cells by therapeutic vaccination may be an innovative strategy to overcome virus persistence. Lentiviral vectors developed and optimized by THERAVECTYS, due to their ability to transduce non-dividing cells, including dendritic cells, and induce CMI response, have demonstrated their effectiveness as vaccination tools. Method: To develop a HBV therapeutic vaccine that can induce a broad but specific immune response, we generated recombinant lentiviral vector carrying IRES(Internal Ribosome Entry Site)-containing bicistronic constructs which allow the coexpression of two vaccine products, namely HBV T- cell epitope vaccine and HBV virus like particle (VLP) vaccine. HBV T-cell epitope vaccine consists of immunodominant cluster of CD4 and CD8 epitopes with spacer in between them and epitopes are derived from HBV surface protein, HBV core, HBV X and polymerase. While HBV VLP vaccine is a HBV core protein based chimeric VLP with surface protein B-cell epitopes displayed. In order to evaluate the immunogenicity, mice were immunized with lentiviral constructs by intramuscular injection. The T cell and antibody immune responses of the two vaccine products were analyzed using IFN-γ ELISpot assay and ELISA respectively to quantify the adaptive response to HBV antigens. Results: Following a single administration in mice, lentiviral construct elicited robust antigen-specific IFN-γ responses to the encoded antigens. The HBV T- cell epitope vaccine demonstrated significantly higher T cell immunogenicity than HBV VLP vaccine. Importantly, we demonstrated by ELISA that antibodies are induced against both HBV surface protein and HBV core protein when mice injected with vaccine construct (p < 0.05). Conclusion: Our results highlight that THERAVECTYS lentiviral vectors may represent a powerful platform for immunization strategy against chronic hepatitis B. Our data suggests the likely importance of Lentiviral vector based novel bicistronic construct for further study, in combination with drugs or as standalone antigens, as a therapeutic lentiviral based HBV vaccines. THERAVECTYS bicistronic HBV vaccine will be further evaluated in animal efficacy studies.

Keywords: chronic hepatitis B, lentiviral vectors, therapeutic vaccine, virus-like particle

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Authors: Ram Sharma, Esha Chatterjee, Santosh Kumar Guru, Kunal Nepali

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A tractable anti-lung cancer agent was identified via the installation of a Ring C expanded synthetic analogue of the alkaloid vasicinone [7,8,9,10-tetrahydroazepino[2,1-b] quinazolin-12(6H)-one (TAZQ)] as a surface recognition part in the HDAC inhibitory three-component model. Noteworthy to mention that the candidature of TAZQ was deemed suitable for accommodation in HDAC inhibitory pharmacophore as per the results of the fragment recruitment process conducted by our laboratory. TAZQ was pinpointed through the fragment screening program as a synthetically flexible fragment endowed with some moderate cell growth inhibitory activity against the lung cancer cell lines, and it was anticipated that the use of the aforementioned fragment to generate hydroxamic acid functionality (zinc-binding motif) bearing HDAC inhibitors would boost the antitumor efficacy of TAZQ. Consistent with our aim of applying epigenetic targets to the treatment of lung cancer, a strikingly potent anti-lung cancer scaffold (compound 6) was pinpointed through a series of in-vitro experiments. Notably, the compounds manifested a magnificent activity profile against KRAS and EGFR mutant lung cancer cell lines (IC50 = 0.80 - 0.96 µM), and the effects were found to be mediated through preferential HDAC6 inhibition (IC50 = 12.9 nM). In addition to HDAC6 inhibition, the compounds also elicited HDAC1 and HDAC3 inhibitory activity with an IC50 value of 49.9 nM and 68.5 nM, respectively. The HDAC inhibitory ability of compound 6 was also confirmed from the results of the western blot experiment that revealed its potential to decrease the expression levels of HDAC isoforms (HDAC1, HDAC3, and HDAC6). Noteworthy to mention that complete downregulation of the HDAC6 isoform was exerted by compound 6 at 0.5 and 1 µM. Moreover, in another western blot experiment, treatment with hydroxamic acid 6 led to upregulation of H3 acK9 and α-Tubulin acK40 levels, ascertaining its inhibitory activity toward both the class I HDACs and Class II B HDACs. The results of other assays were also encouraging as treatment with compound 6 led to the suppression of the colony formation ability of A549 cells, induction of apoptosis, and increase in autophagic flux. In silico studies led us to rationalize the results of the experimental assay, and some key interactions of compound 6 with the amino acid residues of HDAC isoforms were identified. In light of the impressive activity spectrum of compound 6, a pH-responsive nanocarrier (hyaluronic acid-compound 6 nanoparticles) was prepared. The dialysis bag approach was used for the assessment of the nanoparticles under both normal and acidic circumstances, and the pH-sensitive nature of hyaluronic acid-compound 6 nanoparticles was confirmed. Delightfully, the nanoformulation was devoid of cytotoxicity against the L929 mouse fibroblast cells (normal settings) and exhibited selective cytotoxicity towards the A549 lung cancer cell lines. In a nutshell, compound 6 appears to be a promising adduct, and a detailed investigation of this compound might yield a therapeutic for the treatment of lung cancer.

Keywords: HDAC inhibitors, lung cancer, scaffold, hyaluronic acid, nanoparticles

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Authors: Ruolan Zhang, Ryo Imai, Naoko Senda, Tomoyuki Sakai

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Enzymes have attracted increasing attentions in industrial manufacturing for their applicability in catalyzing complex chemical reactions under mild conditions. Directed evolution has become a powerful approach to optimize enzymes and exploit their full potentials under the circumstance of insufficient structure-function knowledge. With the incorporation of cell-free synthetic biotechnology, rapid enzyme synthesis can be realized because no cloning procedure such as transfection is needed. Its open environment also enables direct enzyme measurement. These properties of cell-free biotechnology lead to excellent throughput of enzymes generation. However, the capabilities of current screening methods have limitations. Fluorescence-based assay needs applicable fluorescent label, and the reliability of acquired enzymatic activity is influenced by fluorescent label’s binding affinity and photostability. To acquire the natural activity of an enzyme, another method is to combine pre-screening step and high-performance liquid chromatography (HPLC) measurement. But its throughput is limited by necessary time investment. Hundreds of variants are selected from libraries, and their enzymatic activities are then identified one by one by HPLC. The turn-around-time is 30 minutes for one sample by HPLC, which limits the acquirable enzyme improvement within reasonable time. To achieve the real high-throughput enzyme screening, i.e., obtain reliable enzyme improvement within reasonable time, a widely applicable high-throughput measurement of enzymatic reactions is highly demanded. Here, a high-throughput screening method using broadband coherent anti-Stokes Raman spectroscopy (CARS) was proposed. CARS is one of coherent Raman spectroscopy, which can identify label-free chemical components specifically from their inherent molecular vibration. These characteristic vibrational signals are generated from different vibrational modes of chemical bonds. With the broadband CARS, chemicals in one sample can be identified from their signals in one broadband CARS spectrum. Moreover, it can magnify the signal levels to several orders of magnitude greater than spontaneous Raman systems, and therefore has the potential to evaluate chemical's concentration rapidly. As a demonstration of screening with CARS, alcohol dehydrogenase, which converts ethanol and nicotinamide adenine dinucleotide oxidized form (NAD+) to acetaldehyde and nicotinamide adenine dinucleotide reduced form (NADH), was used. The signal of NADH at 1660 cm⁻¹, which is generated from nicotinamide in NADH, was utilized to measure the concentration of it. The evaluation time for CARS signal of NADH was determined to be as short as 0.33 seconds while having a system sensitivity of 2.5 mM. The time course of alcohol dehydrogenase reaction was successfully measured from increasing signal intensity of NADH. This measurement result of CARS was consistent with the result of a conventional method, UV-Vis. CARS is expected to have application in high-throughput enzyme screening and realize more reliable enzyme improvement within reasonable time.

Keywords: Coherent Anti-Stokes Raman Spectroscopy, CARS, directed evolution, enzyme screening, Raman spectroscopy

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Authors: Z. Nurzaireena, K. Azalea, T. Azirawaty, S. Jameela, G. Muralitharan

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The aim of this study is the review of the management practices of sickle cell disease patients during pregnancy, as well as the maternal and neonatal outcome at Ampang Hospital, Selangor. The study consisted of a review of pregnant patients with sickle cell disease under follow up at the Hematology Clinic, Ampang Hospital over the last seven years to assess their management and maternal-fetal outcome. The results of the review show that Ampang Hospital is considered the public hematology centre for sickle cell disease and had successfully managed three pregnancies throughout the last seven years. Patients’ presentations, managements and maternal-fetal outcome were compared and reviewed for academic improvements. All three patients were seen very early in their pregnancy and had been given a regime of folic acid, antibiotics and thrombo-prophylactic drugs. Close monitoring of maternal and fetal well being was done by the hematologists and obstetricians. Among the patients, there were multiple admissions during the pregnancy for either a painful sickle cell bone crisis, haemolysis following an infection and anemia requiring phenotype- matched blood and exchange transfusions. Broad spectrum antibiotics coverage during and infection, hydration, pain management and venous-thrombolism prophylaxis were mandatory. The pregnancies managed to reach near term in the third trimester but all required emergency caesarean section for obstetric indications. All pregnancies resulted in live births with good fetal outcome. During post partum all were nursed closely in the high dependency units for further complications and were discharged well. Post partum follow up and contraception counseling was comprehensively given for future pregnancies. Sickle cell disease is uncommonly seen in the East, especially in the South East Asian region, yet more cases are seen in the current decade due to improved medical expertise and advance medical laboratory technologies. Pregnancy itself is a risk factor for sickle cell patients as increased thrombosis event and risk of infections can lead to multiple crisis, haemolysis, anemia and vaso-occlusive complications including eclampsia, cerebrovasular accidents and acute bone pain. Patients mostly require multiple blood product transfusions thus phenotype-matched blood is required to reduce the risk of alloimmunozation. Emphasizing the risks and complications in preconception counseling and establishing an ultimate pregnancy plan would probably reduce the risk of morbidity and mortality to the mother and unborn child. Early management for risk of infection, thromboembolic events and adequate hydration is mandatory. A holistic approach involving multidisciplinary team care between the hematologist, obstetricians, anesthetist, neonatologist and close nursing care for both mother and baby would ensure the best outcome. In conclusion, sickle cell disease by itself is a high risk medical condition and pregnancy would further amplify the risk. Thus, close monitoring with combine multidisciplinary care, counseling and educating the patients are crucial in achieving the safe outcome.

Keywords: anaemia, haemoglobinopathies, pregnancy, sickle cell disease

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Authors: Olga Smilevska Spasova, Katerina Boshkovska, Gorica Popova, Mirjana Popovska

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Introduction: Pneumonia is an infection of the pulmonary parenchyma. HPIV 3 is one of four viruses that is a member of the Paramyxoviridae family designated types 1-4 that have a nonsegmented, single-stranded RNA genome with a lipid-containing envelope. They are spread from the respiratory tract by aerosolized secretions or by direct contact with secretions. Type 3 is endemic and can cause serious illness in immunocompromised patients. Illness caused by parainfluenza occurs shortly after inoculation with the virus. The level of immunoglobulin A antibody in serum is the best predictor of susceptibility to infection. Streptococcus pneumonia or pneumococcus is a Gram-positive, spherical bacteria, usually found in pairs and it is a member of the genus Streptococcus. Streptococcus pneumonia resides asymptomatically in healthy carriers typically colonizing the respiratory tract, sinuses, and nasal cavity. In individuals with weaker immune systems like young infants, pneumococcal bacterium is the most common cause of community-acquired pneumonia in the world. Case Report: The aim is to present a case of lower respiratory tract infection in an infant caused by parainfluenza virus 3, S. pneumonia and undifferentiated gram-negative bacteria that was successfully treated. The infant is with a history of recurrent episodes of wheezing in the past 3mounts.Infant of 10months presents 2weeks before admittance with high fever, runny nose, and cough. The primary pediatrician prescribed oral cefpodoxime for 10days and inhaled salbutamol. Two days before admittance in hospital the infant with high fever, cough, and difficulty breathing. At admittance, infant is pale, anxious with rapid respirations, cough, wheezing and tachycardia. On auscultation: vesicular breathing sounds with high pitched wheezing and on the right coarse crackles. Investigations: Blood analysis: RBC: 4, 7 x1012L, WBC: 8,3x109L: Neut: 42.73% Lym: 41.57%, Hgb: 9.38 g/dl MCV: 62.7fl, MCH: 20.0pg MCHC: 31.8 g/dl RDW: 18.7% Plt-307.9 x109LCRP: 2,5mg/l, serum iron-7.92umol/l, O2sat-97% on blood gas analysis, puls-125/min.X-ray of chest with hyperinflationand right pericardial consolidation. Microbiological analysis of sputum sample is positive for undifferentiated gram-negative bacteria (colonizer)–resistant to cefotaxime, ampicillin, cefoxitin, sulfamet.+trimetoprim and sensitive to amikacin, gentamicin, and ciprofloxacin. Molecular multiplex RT-PCR for 19 viruses and multiplex PCR for 7 bacteria test for respiratory pathogens positive for Parainfluenza virus 3(Ct=22.73), Streptococcus pneumonia (Ct=26.75).IED: IgG-9.31g/l, IgA-0.351g/l, IgM-0.86g/l. Therapy: Treatment was started with inhaled salbutamol, intravenous antibiotic cefotaxime as well as systemic corticosteroids. On day 7 because of slow clinical resolution of chest auscultation findings and an etiologic clue with a positive sputum sample for resistant undifferentiated gram negative bacteria, a second intravenous antibiotic was administered amikacin. The infant is discharged on day 14 with resolution of clinical findings. Conclusion: Mixed co-infections with respiratory viruses and bacteria in immunocompromised infants are likely to lead to a more severe form of community acquired pneumonia that will need hospitalization.

Keywords: HPIV- 3, infant, pneumonia, S. pneumonia, x-ray chest

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Authors: Kaushikk Iyer, Richard M Hall, David Keeling

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Hardware-in-the-loop (HIL) simulation is an advanced technique for developing and testing complex real-time control systems. This paper presents the benefits of HIL simulation and how it can be implemented and used effectively to develop, test, and validate advanced control algorithms used in a spinal joint Wear simulator for the Tribological testing of spinal disc prostheses. spinal wear simulator is technologically the most advanced machine currently employed For the in-vitro testing of newly developed spinal Discimplants. However, the existing control techniques, such as a simple position control Does not allow the simulator to test non-sinusoidal waveforms. Thus, there is a need for better and advanced control methods that can be developed and tested Rigorouslybut safely before deploying it into the real simulator. A benchtop HILsetupis was created for experimentation, controller verification, and validation purposes, allowing different control strategies to be tested rapidly in a safe environment. The HIL simulation aspect in this setup attempts to replicate similar spinal motion and loading conditions. The spinal joint wear simulator containsa four-Barlinkpowered by electromechanical actuators. LabVIEW software is used to design a kinematic model of the spinal wear Simulator to Validatehow each link contributes towards the final motion of the implant under test. As a result, the implant articulates with an angular motion specified in the international standards, ISO-18192-1, that define fixed, simplified, and sinusoid motion and load profiles for wear testing of cervical disc implants. Using a PID controller, a velocity-based position control algorithm was developed to interface with the benchtop setup that performs HIL simulation. In addition to PID, a fuzzy logic controller (FLC) was also developed that acts as a supervisory controller. FLC provides intelligence to the PID controller by By automatically tuning the controller for profiles that vary in amplitude, shape, and frequency. This combination of the fuzzy-PID controller is novel to the wear testing application for spinal simulators and demonstrated superior performance against PIDwhen tested for a spectrum of frequency. Kaushikk Iyer is a Ph.D. Student at the University of Leeds and an employee at Key Engineering Solutions, Leeds, United Kingdom, (e-mail: [email protected], phone: +44 740 541 5502). Richard M Hall is with the University of Leeds, the United Kingdom as a professor in the Mechanical Engineering Department (e-mail: [email protected]). David Keeling is the managing director of Key Engineering Solutions, Leeds, United Kingdom (e-mail: [email protected]). Results obtained are successfully validated against the load and motion tolerances specified by the ISO18192-1 standard and fall within limits, that is, ±0.5° at the maxima and minima of the motion and ±2 % of the complete cycle for phasing. The simulation results prove the efficacy of the test setup using HIL simulation to verify and validate the accuracy and robustness of the prospective controller before its deployment into the spinal wear simulator. This method of testing controllers enables a wide range of possibilities to test advanced control algorithms that can potentially test even profiles of patients performing various dailyliving activities.

Keywords: Fuzzy-PID controller, hardware-in-the-loop (HIL), real-time simulation, spinal wear simulator

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Authors: Raham El-Kahlout, Hadi Yassin, Asmaa Athani, Marwan Abou Madi, Gheyath Nasrallah

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Background: Since its first isolation in September 2012, Middle East respiratory syndrome coronavirus (MERS-CoV) has diffused across 27 countries infecting more than two thousand individuals with a high case fatality rate. MERS-CoV–specific antibodies are widely found in Dromedary camel along with viral shedding of similar viruses detected in human at same region, suggesting that MERS epidemiology may be central role by camel. Interestingly, MERS-CoV has also been also reported to be asymptomatic or to cause influenza-like mild illnesses. Therefore, in a country like Qatar (bordered Saudi Arabia), where camels are widely spread, serological surveys are important to explore the role of camels in MERS-CoV transmission. However, widespread strategic serological surveillances of MERS-CoV among populations, particularly in endemic country, are infrequent. In the absence of clear epidemiological view, cross-sectional MERS antibody surveillances in human populations are of global concern. Method: We performed a comparative serological screening of 4719 healthy blood donors, 135 baseline case contacts (high risk individual), and four MERS confirmed patients (by PCR) for the presence of anti-MERS IgG. Initially, samples were screened using Euroimmune anti- MERS-CoV IgG ELISA kit, the only commercial kit available in the market and recommended by the CDC as a screening kit. To confirm ELISA test results, farther serological testing was performed for all borderline and positive samples using two assays; the anti MERS-CoV IgG and IgM Euroimmune indirect immunofluorescent test (IIFT) and pseudoviral particle neutralizing assay (PPNA). Additionally, to test cross reactivity of anti-MERS-CoV antibody with other family members of coronavirus, borderline and positive samples were tested for the presence of the of IgG antibody of the following viruses; SARS, HCoV-229E, HKU1 using the Euroimmune IIFT for SARS and HCoV-229E and ELISA for HKU1. Results: In all of 4858 screened 15 samples [10 donors (0.21%, 10/4719), 1 case contact (0.77 %, 1/130), 3 patients (75%, 3/4)] anti-MERS IgG reactive/borderline samples were seen in ELISA. However, only 7 (0.14%) of them gave positive with in IIFT and only 3 (0.06%) was confirmed by the specific anti-MERS PPNA. One of the interesting findings was, a donor, who was selected in the control group as a negative anti-MERS IgG ELISA, yield reactive for anti-MERS IgM IIFT and was confirmed with the PPNA. Further, our preliminary results showed that there was a strong cross reactivity between anti- MERS-COV IgG with both HCoV-229E or anti-HKU1 IgG, yet, no cross reactivity of SARS were found. Conclusions: Our findings suggest that MERS-CoV is not heavily circulated among the population of Qatar and this is also indicated by low number of confirmed cases (only 18) since 2012. Additionally, the presence of antibody of other pathogenic human coronavirus may cause false positive results of both ELISA and IIFT, which stress the need for more evaluation studies for the available serological assays. Conclusion: this study provides an insight about the epidemiological view for MERS-CoV in Qatar population. It also provides a performance evaluation for the available serologic tests for MERS-CoV in a view of serologic status to other human coronaviruses.

Keywords: seroprevalence, MERS-CoV, healthy individuals, Qatar

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Authors: LouAnne Boyd

Abstract:

From the perspective of self-advocates, the biggest unaddressed problem is not the symptoms of an autism spectrum diagnosis but the social stigma that accompanies autism. This societal perspective is in contrast to the focus on the majority of interventions. Autism interventions, and consequently, most innovative technologies for autism, aim to improve deficits that occur within the person. For example, the most common Human-Computer Interaction research projects in assistive technology for autism target social skills from a normative perspective. The premise of the autism technologies is that difficulties occur inside the body, hence, the medical model focuses on ways to improve the ailment within the person. However, other technological approaches to support people with autism do exist. In the realm of Human Computer Interaction, there are other modes of research that provide critique of the medical model. For example, critical design, whose intended audience is industry or other HCI researchers, provides products that are the opposite of interventionist work to bring attention to the misalignment between the lived experience and the societal perception of autism. For example, parodies of interventionist work exist to provoke change, such as a recent project called Facesavr, a face covering that helps allistic adults be more independent in their emotional processing. Additionally, from a critical disability studies’ perspective, assistive technologies perpetuate harmful normalizing behaviors. However, these critical approaches can feel far from the frontline in terms of taking direct action to positively impact end users. From a critical yet more pragmatic perspective, projects such as Counterventions lists ways to reduce the likelihood of perpetuating ableism in interventionist’s work by reflectively analyzing a series of evolving assistive technology projects through a societal lens, thus leveraging the momentum of the evolving ecology of technologies for autism. Therefore, all current paradigms fall short of addressing the largest need—the negative impact of social stigma. The current work introduces a new paradigm for technologies for autism, borrowing from a paradigm introduced two decades ago around changing the narrative related to eating disorders. It is the shift from reprimanding poor habits to celebrating positive aspects of eating. This work repurposes Celebratory Technology for Neurodiversity and intended to reduce social stigma by targeting for the public at large. This presentation will review how requirements were derived from current research on autism social stigma as well as design sessions with autistic adults. Congruence between these two sources revealed three key design implications for technology: provide awareness of the autistic experience; generate acceptance of the neurodivergence; cultivate an appreciation for talents and accomplishments of neurodivergent people. The current pilot work in Celebratory Technology offers a new paradigm for supporting autism by shifting the burden of change from the person with autism to address changing society’s biases at large. Shifting the focus of research outside of the autistic body creates a new space for a design that extends beyond the bodies of a few and calls on all to embrace humanity as a whole.

Keywords: neurodiversity, social stigma, accessibility, inclusion, celebratory technology

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Authors: Siddharudha B. Patil

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Cotton is a premier commercial fibre crop of India subjected to ravages of insect pests. Sucking pests viz thrips, Thrips tabaci,(lind) leaf hopper Amrsca devastance,(dist) miridbug, Poppiocapsidea beseratense (Dist) and bollworms continue to inflict damage Bt Cotton right from seeding stage. Their infestation impact cotton yield to an extent of 30-40 percent. Chemical control is still adoptable as one of the techniques for combating these pests. Presently, growers have many challenges in selecting effective chemicals which fit in with an integrated pest management. Spinetoram has broad spectrum with excellent insecticidal activity against both sucking pests and bollworms. Hence, it is expected to make a great contribution to stable production and quality improvement of agricultural products. Spinetoram is a derivative of biologically active substances (Spinosyns) produced by soil actinomycetes, Saccharopolypara spinosa which is semi synthetic active ingredient representing Spinosyn chemical class of insecticide and has demonstrated higher level of efficacy with reduced risk on beneficial arthropods. The efforts were made in the present study to test the efficacy of Spinetoram against sucking pests and bollworms in comparison with other insecticides in Bt Cotton under field condition. Field experiment was laid out during 2013-14 and 2014-15 at Agricultural Research station Dharwad (Karnataka-India) in a randomized block design comprising eight treatments and three replications. Bt cotton genotype, Bunny BG-II was sown in a plot size of 5.4 m x5.4 m. Recommend agronomical practices were followed. The Spinetoram 12% SC alone and incombination with sulfaxaflore with varied dosages against pest complex was tested. Performance was compared with Spinosad 45% SC and thiamethoxam 25% WG. The results of consecutive seasons revealed that nonsignificant difference in thrips and leafhopper population and varied significantly after 3 days of imposition. Among the treatments, combiproduct, Spinetoram 10%WG + Sulfoxaflor 30% WG@ 140 gai/ha registered lowest population of thrips (3.91/3 leaves) and leaf hoppers (1.08/3 leaves) followed by its lower dosages viz 120 gai/ha (4.86/3 leaves and 1.14/3 leaves of thrips and leaf hoppers, respectively) and 100 gai/ha (6.02 and 1.23./3 leaves of thrips and leaf hoppers respectively) being at par, significantly superior to rest of the treatments. On the contrary, the population of thrips, leaf hopper and miridbugs in untreated control was on higher side. Similarly the higher dosage of Spinetoram 10% WG+ Sulfoxaflor 30% WG (140 gai/ha) proved its bioefficacy by registering lowest miridbug incidence of 1.70/25 squares, followed by its lower dosage (1.78 and 1.83/25 squares respectively) Further observation made on bollworms incidence revealed that the higher dosage of Spinetoram 10% WG+Sulfoxaflor 30% WG (140 gai/ha) registered lowest percentage of boll damage (7.22%), more number of good opened bolls (36.89/plant) and higher seed cotton yield (19.45q/ha) followed by rest of its lower dosages, Spinetoram 12% SC alone and Spinosad 45% SC being at par significantly superior to rest of the treatments. However, significantly higher boll damage (15.13%) and lower seed cotton yield (14.45 q/ha) was registered in untreated control. Thus Spinetoram10% WG+Sulfoxaflor 30% WG can be a promising option for pest management in Bt Cotton.

Keywords: Spinetoram10% WG+Sulfoxaflor 30% WG, sucking pests, bollworms, Bt cotton, management

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Authors: Saeed Abbas, Rahilla Tabassum, Haider Abbas, Babar Khan, Shahid Hussain, Muhammad Zafar Khan, Fazal Karim, Yawar Abbas, Rizwan Karim

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Human–carnivore conflict is considered to be a major conservation and rural livelihood concern because many carnivore species have been heavily victimized due to elevated conflict levels with communities. Like other snow leopard range countries, this situation prevails in Pakistan, where WWF is currently working under Asia High Mountain Project (AHMP) in Gilgit-Baltistan of Pakistan. To mitigate such conflicts requires a firm understanding of grazing and predation pattern including human-carnivore interaction. For this purpose we conducted a survey in Hoper valley (one of the AHMP project sites in Pakistan), during August, 2013 through a questionnaire based survey and unstructured interviews covering 647 households, permanently residing in the project area out of the total 900 households. The valley, spread over 409 km2 between 36°7'46" N and 74°49'2"E, at 2900m asl in Karakoram ranges is considered to be one of an important habitat of snow leopard and associated prey species such as Himalayan ibex. The valley is home of 8100 Brusho people (ancient tribe of Northern Pakistan) dependent on agro-pastoral livelihoods including farming and livestock rearing. The total number of livestock reported were (N=15,481) out of which 8346 (53.91%) were sheep, 3546 (22.91%) goats, 2193 (14.16%) cows, 903 (5.83%) yaks, 508 (3.28%) bulls, 28 (0.18%) donkeys, 27 (0.17%) zo/zomo (cross breed of yak and cow), and 4 (0.03%) horses. 83 percent respondent (n=542 households) confirmed loss of their livestock during the last one year July, 2012 to June, 2013 which account for 2246 (14.51%) animals. The major reason of livestock loss include predation by large carnivores such as snow leopards and wolf (1710, 76.14%) followed by diseases (536, 23.86%). Of the total predation cases snow leopard is suspected to kill 1478 animals (86.43%). Among livestock sheep were found to be the major prey of snow leopard (810, 55%) followed by goats (484, 32.7%) cows (151, 10.21%), yaks (15, 1.015%), zo/zomo (7, 0.5%) and donkey (1, 0.07%). The reason for the mass depredation of sheep and goats is that they tend to browse on twigs of bushes and graze on soft grass near cliffs. They are also considered to be very active as compared to other species in moving quickly and covering more grazing area. This makes them more vulnerable to snow leopard attack. The majority (1283, 75%) of livestock killed by predators occurred during the warm season (May-September) in alpine and sub-alpine pastures and remaining (427, 25%) occurred in the winter season near settlements in valley. It was evident from the recent study that Snow leopard kills outside the pen were (1351, 79.76%) as compared to inside pen (359, 20.24%). Assessing the economic loss of livestock predation we found that the total loss of livestock predation in the study area is equal to PKR 11,230,000 (USD 105,797), which is about PRK 17, 357 (USD 163.51) per household per year. Economic loss incurred by the locals due to predation is quite significant where the average cash income per household per year is PKR 85,000 (USD 800.75).

Keywords: carnivores, conflict, predation, livelihood, conservation, rural, snow leopard, livestock

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Authors: Prem Kumar, Anuj Tyagi, Harsh Panwar, Vaneet Inder Kaur

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Though aquaculture started as an occupation for poor and weak farmers for livelihood, it has now acquired the shape of one of the biggest industry to grow live protein in the form of aquatic organisms. Industrialization of the aquaculture sector has led to intensification resulting in stress on aquatic organisms and frequent disease outbreaks leading to huge economic impacts. Indiscriminate use of antibiotics as growth promoter and prophylactic agent in aquaculture has resulted in rapid emergence and spread of antibiotic resistance in bacterial pathogens. Over the past few years, use of probiotics (as an alternative of antibiotics) in aquaculture has gained attention due to their immunostimulant and growth promoting properties. It has now well known that after administration, a probiotic bacterium has to compete and establish itself against native microbiota to show its eventual beneficial properties. Due to their non-fish origin, commercial probiotics sometimes may display poor probiotic functionalities and antagonistic effects. Thus, isolation and characterization of probiotic bacteria from same fish host is very much necessary. In this study, attempts were made to isolate potent probiotic lactic acid bacteria (LAB) from intestinal microflora of rohu fish. Twenty-five experimental rohu fishes (mean weight 400 ± 20gm, mean standard length 20 ± 3cm) were used in the study to collect fish gut after dissection in a sterile condition. A total of 150 tentative LAB isolates from selective agar media (de Man-Rogosa-Sharpe (MRS)) were screened for their antimicrobial activity against Aeromonas hydrophila and Microccocus leuteus. A total of 17 isolates, identified as Lactobacillus plantarum and Lactococcus lactis, identified by biochemical tests and PCR amplification and sequencing of 16S rRNA gene fragment, displayed promising antimicrobial activity against both the pathogens. Two isolates from each species (FLB1, FLB2 from L. plantarum; and FLC1, FLC2 from L. lactis) were subjected to downstream probiotic potential characterization. These isolates were compared in vitro for their hemolytic activity, acid and bile tolerance for growth kinetics, auto-aggregation, cell-surface hydrophobicity against xylene, and chloroform, tolerance to phenol, cell adhesion, and safety parameters (by intraperitoneal and intramuscular injections). None of the tested isolates showed any hemolytic activity indicating their potential safety. Moreover, these isolates were tolerant to 0.3% bile (75-82% survival), phenol stress (96-99% survival) with 100% viability at pH 3 over a period of 3 h. Antibiotic sensitivity test revealed that all the tested LAB isolates were resistant to vancomycin, gentamicin, streptomycin, and erythromycin and sensitive to Erythromycin, Chloramphenicol, Ampicillin, Trimethoprim, and Nitrofurantoin. Tetracycline resistance was found in L. plantarum (FLB1 and FLB2 isolates), whereas L. lactis were susceptible to it. Intramuscular and intraperitoneal challenges to fingerlings of rohu fish (5 ± 1gm weight) with FLB1 showed no pathogenicity and occurrence of disease symptoms in fishes over an observation period of 7 days. The results revealed FLB1 as a potential probiotic candidate for aquaculture application among other isolates.

Keywords: aquaculture, Lactobacillus plantarum, Lactococcus lactis, probiotics

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Authors: Ting Wu, Feiting Hu, Xinyue Zhang, Shuxin Tang, Xiaoyun Xu

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As a by-product of yuba, processing wastewater of Yuba (PWY) contains many bioactive components such as soybean isoflavones, soybean polysaccharides and soybean oligosaccharides, which is a good source of prebiotics and has a potential of high value utilization. The use of Lactobacillus plantarum to ferment PWY can be considered as a potential biogenic element, which can regulate the balance of intestinal microbiota. In this study, firstly, Lactobacillus plantarum was used to ferment PWY to improve its content of active components and antioxidant activity. Then, the health effect of fermented processing wastewater of yuba (FPWY) was measured in vitro. Finally, microencapsulation technology was used applied to improve the sustained release of FPWY and reduce the loss of active components in the digestion process, as well as to improving the activity of FPWY. The main results are as follows: (1) FPWY presented a good antioxidant capacity with DPPH free radical scavenging ability (0.83 ± 0.01 mmol Trolox/L), ABTS free radical scavenging ability (7.47 ± 0.35 mmol Trolox/L) and iron ion reducing ability (1.11 ± 0.07 mmol Trolox/L). Compared with non-fermented processing wastewater of yuba (NFPWY), there was no significant difference in the content of total soybean isoflavones, but the content of glucoside soybean isoflavones decreased, and aglyconic soybean isoflavones increased significantly. After fermentation, PWY can effectively reduce the soluble monosaccharides, disaccharides and oligosaccharides, such as glucose, fructose, galactose, trehalose, stachyose, maltose, raffinose and sucrose. (2) FPWY can significantly enhance the growth of beneficial bacteria such as Bifidobacterium, Ruminococcus and Akkermansia, significantly inhibit the growth of harmful bacteria E.coli, regulate the structure of intestinal microbiota, and significantly increase the content of short-chain fatty acids such as acetic acid, propionic acid, butyric acid, isovaleric acid. Higher amount of lactic acid in the gut can be further broken down into short chain fatty acids. (3) In order to improve the stability of soybean isoflavones in FPWY during digestion, sodium alginate and chitosan were used as wall materials for embedding. The FPWY freeze-dried powder was embedded by the method of acute-coagulation bath. The results show that when the core wall ratio is 3:1, the concentration of chitosan is 1.5%, the concentration of sodium alginate is 2.0%, and the concentration of calcium is 3%, the embossing rate is 53.20%. In the simulated in vitro digestion stage, the release rate of microcapsules reached 59.36% at the end of gastric digestion and 82.90% at the end of intestinal digestion. Therefore, the core materials with good sustained-release performance of microcapsules were almost all released. The structural analysis results of FPWY microcapsules show that the microcapsules have good mechanical properties. Its hardness, springness, cohesiveness, gumminess, chewiness and resilience were 117.75± 0.21 g, 0.76±0.02, 0.54±0.01, 63.28±0.71 g·sec, 48.03±1.37 g·sec, 0.31±0.01, respectively. Compared with the unembedded FPWY, the infrared spectrum results showed that the microcapsules had embedded effect on the FPWY freeze-dried powder.

Keywords: processing wastewater of yuba, lactobacillus plantarum, intestinal microbiota, microcapsule

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Authors: Cai J. T. Ladd, Mollie F. Duggan-Edwards, Tjeerd J. Bouma, Jordi F. Pages, Martin W. Skov

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Salt marshes are valued for their role in coastal flood protection, carbon storage, and for supporting biodiverse ecosystems. As a biogeomorphic landscape, marshes evolve through the complex interactions between sea level rise, sediment supply and wave/current forcing, as well as and socio-economic factors. Climate change and direct human modification could lead to a global decline marsh extent if left unchecked. Whilst the processes of saltmarsh erosion and expansion are well understood, empirical evidence on the key drivers of long-term lateral marsh dynamics is lacking. In a GIS, saltmarsh areal extent in 25 estuaries across Great Britain was calculated from historical maps and aerial photographs, at intervals of approximately 30 years between 1846 and 2016. Data on the key perceived drivers of lateral marsh change (namely sea level rise rates, suspended sediment concentration, bedload sediment flux rates, and frequency of both river flood and storm events) were collated from national monitoring centres. Continuous datasets did not extend beyond 1970, therefore predictor variables that best explained rate change of marsh extent between 1970 and 2016 was calculated using a Partial Least Squares Regression model. Information about the spread of Spartina anglica (an invasive marsh plant responsible for marsh expansion around the globe) and coastal engineering works that may have impacted on marsh extent, were also recorded from historical documents and their impacts assessed on long-term, large-scale marsh extent change. Results showed that salt marshes in the northern regions of Great Britain expanded an average of 2.0 ha/yr, whilst marshes in the south eroded an average of -5.3 ha/yr. Spartina invasion and coastal engineering works could not explain these trends since a trend of either expansion or erosion preceded these events. Results from the Partial Least Squares Regression model indicated that the rate of relative sea level rise (RSLR) and availability of suspended sediment concentration (SSC) best explained the patterns of marsh change. RSLR increased from 1.6 to 2.8 mm/yr, as SSC decreased from 404.2 to 78.56 mg/l along the north-to-south gradient of Great Britain, resulting in the shift from marsh expansion to erosion. Regional differences in RSLR and SSC are due to isostatic rebound since deglaciation, and tidal amplitudes respectively. Marshes exposed to low RSLR and high SSC likely leads to sediment accumulation at the coast suitable for colonisation by marsh plants and thus lateral expansion. In contrast, high RSLR with are likely not offset deposition under low SSC, thus average water depth at the marsh edge increases, allowing larger wind-waves to trigger marsh erosion. Current global declines in sediment flux to the coast are likely to diminish the resilience of salt marshes to RSLR. Monitoring and managing suspended sediment supply is not common-place, but may be critical to mitigating coastal impacts from climate change.

Keywords: lateral saltmarsh dynamics, sea level rise, sediment supply, wave forcing

Procedia PDF Downloads 111