Scholarly Research Excellence

Digital Open Science Index

Commenced in January 2007 Frequency: Monthly Edition: International Abstract Count: 56062

Bioengineering and Life Sciences

Survey of Potato Viral Infection Using Double Antibody Sandwich-Enzyme Linked Immunosorbent Assay Method in Georgia
Plant viruses can cause loss of yield and quality in a lot of important crops. Symptoms of pathogens are variable depending on the cultivars and virus stain. Selection resistant potato varieties would reduce the risk of virus transmission and significant economic impact. Other way to avoid reduced harvest yields is regular potato seed production sampling and testing for viral infection. The aim of this study was to determine the occurrence and distribution of viral diseases according to potato cultivars for further selection of virus-free material in Georgia. During the summer 2015-2016, 5 potato cultivars (Sante, Laura, Jelly, Red Sonia, Anushka) at 5 different farms located in Akhalkalaki were tested for 6 different potato viruses: Potato virus A (PVA), M (PVM), S (PVS), X (PVX), Y (PVY) and potato leaf roll (PLRV). A serological method, Double Antibody Sandwich-Enzyme linked Immunosorbent Assay (DAS-ELISA) was used at the laboratory to analyze the results. The result showed that PVY and PLRV virus presence respectively: 21.4% and 19.7% in collected samples was relatively high comparing to others. Researched potato cultivars except Jelly and Laura were infected by PVY with different concentration. PLRV was found only in three potato cultivars (Sante, Jelly, Red Sonia). With low prevalence was (3.12%) characterized PVM virus. PVX, PVA and PVS virus infection was not reported. It would be noted that 7.9 % of patterns were containing PVY/PLRV mix infection. Based on the result it can be concluded that PVY and PLRV infection are dominant in all research cultivars. Therefore significant yield losses are expected. Systematic, long-term work on potato virus control, where relatively frequent replacement of seed-potatoes must be regarded as the most important factor to increase seed productivity.
Arbuscular Mycorrhizal Symbiosis in Trema orientalis: Effect of a Naturally-Occurring Symbiosis Receptor Kinase Mutant Allele
The Trema genus represents a group of fast-growing tropical tree species within the Cannabaceae. Interestingly, five species nested in this lineage -known as Parasponia- can establish rhizobium nitrogen-fixing root nodules, similar to those found in legumes. Parasponia and legumes use a conserved genetic network to control root nodule formation, among which are genes also essential for mycorrhizal symbiosis (the so-called common symbiotic pathway). However, Trema species lost several genes that function exclusively in nodulation, suggesting a loss-of the nodulation trait in Trema. Strikingly, in a Trema orientalis population found in Malaysian Borneo we identified a truncated SYMBIOSIS RECEPTOR KINASE (SYMRK) mutant allele lacking a large portion of the c-terminal kinase domain. In legumes this gene is essential for nodulation and mycorrhization. This raises the question whether Trema orientalis can still be mycorrhized. To answer this question, we established quantitative mycorrhization assay for Parasponia andersonii and Trema orientalis. Plants were grown in closed pots on half strength Hoagland medium containing 20 µM potassium phosphate in sterilized sand and inoculated with 125 spores of Rhizopagus irregularis (Agronutrion-DAOM197198). Mycorrhization efficiency was determined by analyzing the frequency of mycorrhiza (%F), the intensity of the mycorrhizal colonization (%M) and the arbuscule abundance (%A) in the root system. Trema orientalis RG33 can be mycorrhized, though with lower efficiency compared to Parasponia andersonii. From this we conclude that a functional SYMRK kinase domain is not essential for Trema orientalis mycorrhization. In ongoing experiments, we aim to investigate the role of SYMRK in Parasponia andersonii mycorrhization and nodulation. For this two Parasponia andersonii symrk CRISPR-Cas9 mutant alleles were created. One mimicking the TorSYMRKRG33 allele by deletion of exon 13-15, and a full Parasponia andersonii SYMRK knockout.
Insect Diversity Assessment of Maize Crop (Zea mays L.) by Using Sweep Net, Pitfall Trap and Plant Inspection Methods
Maize is known as queen of cereals because of its highest genetic yield potential and multipurpose characteristics in human being and animal diet. Maize crop visited by many major, minor, visitors and sporadic insect pests. This study was conducted during 2014 to evaluate the richness and evenness of these insect pests and their interaction with metrological conditions at University of Agriculture, Faisalabad. In this experiment, two localities were selected; one was treated with pesticide and second was untreated. Maize field visited by many insect pests. Those insect pests were collected by using three collection method: sweep net, pitfall trap and plant inspection. The data was collected weekly interval from August to October and statistically analyzed by using Shannon Index which showed the results of insect pest richness and evenness. The value of Shannon Index was higher with the increase in number of species and abundance of insects. Camponotus nearcticus was most abundant in sweep net and pitfall trap method while Rhopalosiphum maidis was abundant in plant inspection method. Temperature was negatively co-relate with the insect population in all three collection methods while the relative humidity and rainfall had varying results.
Geographic Variation in the Baseline Susceptibility of Helicoverpa armigera (Hubner) (Noctuidae: Lepidoptera) Field Populations to Bacillus thuringiensis Cry Toxins for Resistance Monitoring
The transgenic cotton expressing Bacillus thuringiensis (Bt) provides an effective control of Helicoverpa armigera, a most damaging pest of the cotton crop. However, Bt cotton may not be the optimal solution owing to the selection pressure of Cry toxins. As Bt cotton express the insecticidal proteins throughout the growing seasons, there are the chances of resistance development in the target pests. A regular monitoring and surveillance of target pest’s baseline susceptibility to Bt Cry toxins is crucial for early detection of any resistance development. The present study was conducted to monitor the changes in the baseline susceptibility of the field population of H. armigera to Bt Cry1Ac toxin. The field-collected larval populations were maintained in the laboratory on artificial diet and F1 generation larvae were used for diet incorporated diagnostic studies. The LC₅₀ and MIC₅₀ were calculated to measure the level of resistance of population as a ratio over susceptible population. The monitoring results indicated a significant difference in the susceptibility (LC₅₀) of H. armigera for first, second, third and fourth instar larval populations sampled from different cotton growing areas over the study period 2016-17. The variations in susceptibility among the tested insects depended on the age of the insect and susceptibility decreased with the age of larvae. The overall results show that the average resistant ratio (RR) of all field-collected populations (FSD, SWL, MLT, BWP and DGK) exposed to Bt toxin Cry1Ac ranged from 3.381-fold to 7.381-fold for 1st instar, 2.370-fold to 3.739-fold for 2nd instar, 1.115-fold to 1.762-fold for 3rd instar and 1.141-fold to 2.504-fold for 4th instar, depicting maximum RR from MLT population, whereas minimum RR for FSD and SWL population. The results regarding moult inhibitory concentration of H. armigera larvae (1-4th instars) exposed to different concentrations of Bt Cry1Ac toxin indicated that among all field populations, overall Multan (MLT) and Bahawalpur (BWP) populations showed higher MIC₅₀ values as compared to Faisalabad (FSD) and Sahiwal (SWL), whereas DG Khan (DGK) population showed an intermediate moult inhibitory concentrations. This information is important for the development of more effective resistance monitoring programs. The development of Bt Cry toxins baseline susceptibility data before the widespread commercial release of transgenic Bt cotton cultivars in Pakistan is important for the development of more effective resistance monitoring programs to identify the resistant H. armigera populations.
Preventing Neurodegenerative Diseases by Stabilization of Superoxide Dismutase by Natural Polyphenolic Compounds
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease caused by misfolding and aggregation of Cu, Zn superoxide dismutase (SOD1). The use of small molecules has been shown to stabilize the SOD1 dimer and preventing its dissociation and aggregation. In this study, we employed molecular docking, molecular dynamics simulation and surface plasmon resonance (SPR) to study the interactions between SOD1 and natural polyphenolic compounds. In order to explore the noncovalent interaction between SOD1 and natural polyphenolic compounds, molecular docking and molecular dynamic (MD) simulations were employed to gain insights into the binding modes and free energies of SOD1-polyphenolic compounds. MM/PBSA methods were used to calculate free energies from obtained MD trajectories. The compounds, Hesperidin, Ergosterol, and Rutin showed the excellent binding affinity in micromolar range with SOD1. Ergosterol and Hesperidin have the strongest binding affinity to SOD1 and was subjected to further characterization. Biophysical experiments using Circular Dichroism and Thioflavin T fluorescence spectroscopy results show that the binding of these two compounds can stabilize SOD1 dimer and inhibit the aggregation of SOD1. Molecular simulation results also suggest that these compounds reduce the dissociation of SOD1 dimers through direct interaction with the dimer interface. This study will be helpful to develop other drug-like molecules which may have the effect to reduce the aggregation of SOD1.
Development of Fluorescence Resonance Energy Transfer-Based Nanosensor for Measurement of Sialic Acid in vivo
Sialic acid (5-Acetylneuraminic acid, Neu5Ac) is a common sugar found as a terminal residue on glycoconjugates in many animals. Humans brain and the central nervous system contain the highest concentration of sialic acid (as N-acetylneuraminic acid) where these acids play an important role in neural transmission and ganglioside structure in synaptogenesis. Due to its important biological function, sialic acid is attracting increasing attention. To understand metabolic networks, fluxes and regulation, it is essential to be able to determine the cellular and subcellular levels of metabolites. Genetically-encoded fluorescence resonance energy transfer (FRET) sensors represent a promising technology for measuring metabolite levels and corresponding rate changes in live cells. Taking this, we developed a genetically encoded FRET (fluorescence resonance energy transfer) based nanosensor to analyse the sialic acid level in living cells. Sialic acid periplasmic binding protein (sia P) from Haemophilus influenzae was taken and ligated between the FRET pair, the cyan fluorescent protein (eCFP) and Venus. The chimeric sensor protein was expressed in E. coli BL21 (DE3) and purified by affinity chromatography. Conformational changes in the binding protein clearly confirmed the changes in FRET efficiency. So any change in the concentration of sialic acid is associated with the change in FRET ratio. This sensor is very specific to sialic acid and found stable with the different range of pH. This nanosensor successfully reported the intracellular level of sialic acid in bacterial cell. The data suggest that the nanosensors may be a versatile tool for studying the in vivo dynamics of sialic acid level non-invasively in living cells
Effectiveness Assessment of a Brazilian Larvicide on Aedes Control
The susceptibility status of an insect population to any larvicide depends on several factors such includes genetic constitution, environmental conditions and others. The mosquito Aedes aegypti is the primary vector of three important viral diseases, Zika, Dengue, and Chikungunya. The frequent outbreaks of those diseases in different parts of Brazil demonstrate the importance of testing the susceptibility of vectors in different environments. Since the control of this mosquito leads to the control of disease, alternatives for vector control that value the different Brazilian environmental conditions are needed for effective actions. The aim of this study was to evaluate a new commercial formulation of Bacillus thuringiensis israelenses (DengueTech: Brazilian innovative technology) in the Brazilian Legal Amazon considering the climate conditions. Semi-field tests were conducted in the Institute of Scientific and Technological Research of the State of Amapa in two different environments, one in a shaded area and the other exposed to sunlight. The mosquito larvae were exposed to larvicide concentration and a control; each group was tested in three containers of 40 liters each. To assess persistence 50 third instar larvae of Aedes aegypti laboratory lineages (Rockefeller) and 50 larvae of Aedes aegypti collected in the municipality of Macapa, Brazil’s Amapa state, were added weekly and after 24 hours the mortality was assessed. In total 16 tests were performed, where 12 were done with replacement of water (1/5 of the volume, three times per week). The effectiveness of the product was determined through mortality of ≥ 80%, as recommend by the World Health Organization. The results demonstrated that high-water temperatures (26-35 °C) on the containers influenced the residual time of the product, where the maximum effect achieved was 21 days in the shaded area; and no effectiveness of 60 days was found in any of the tests, as expected according to the larvicide company. The test with and without water replacement did not present significant differences in the mortality rate. Considering the different environments and climate, these results stimulate the need to test larvicide and its effectiveness in specific environmental settings in order to identify the parameters required for better results. Thus, we see the importance of semi-field researches considering the local climate conditions for a successful control of Aedes aegypti.
The Residual Efficacy of Etofenprox WP on Different Surfaces for Malaria Control in the Brazilian Legal Amazon
Malaria is a public health problem in the Brazilian Legal Amazon. Among the integrated approaches for anopheline control, the Indoor Residual Spraying (IRS) remains one of the main tools in the basic strategy applied in the Amazonian States, where the National Malaria Control Program currently uses one of the insecticides from the pyrethroid class, the Etofenprox WP. Understanding the residual efficacy of insecticides on different surfaces is essential to determine the spray cycles, in order to maintain a rational use and to avoid product waste. The aim of this study was to evaluate the residual efficacy of Etofenprox - VECTRON ® 20 WP on surfaces of Unplastered Cement (UC) and Unpainted Wood (UW) on panels, in field, and in semi-field evaluation of Brazil’s Amapa State. The evaluation criteria used was the cone bioassay test, following the World Health Organization (WHO) recommended method, using plastic cones and female mosquitos of Anopheles sp. The tests were carried out in laboratory panels, semi-field evaluation in a “test house” built in the Macapa municipality, and in the field in 20 houses, being ten houses per surface type (UC and UW), in an endemic malaria area in Mazagão’s municipality. The residual efficacy was measured from March to September 2017, starting one day after the spraying, repeated monthly for a period of six months. The UW surface presented higher residual efficacy than the UC. In fact, the UW presented a residual efficacy of the insecticide throughout the period of this study with a mortality rate above 80% in the panels (= 95%), in the "test house" (= 86%) and in field houses ( = 87%). On the UC surface it was observed a mortality decreased in all the tests performed, with a mortality rate of 45, 47 and 29% on panels, semi-field and in field, respectively; however, the residual efficacy ≥ 80% only occurred in the first evaluation after the 24-hour spraying bioassay in the "test house". Thus, only the UW surface meets the specifications of the World Health Organization Pesticide Evaluation Scheme (WHOPES) regarding the duration of effective action (three to six months). To sum up, the insecticide residual efficacy presented variability on the different surfaces where it was sprayed. Although the IRS with Etofenprox WP was efficient on UW surfaces, and it can be used in spraying cycles at 4-month intervals, it is important to consider the diversity of houses in the Brazilian Legal Amazon, in order to implement alternatives for vector control, including the evaluation of new products or different formulations types for insecticides.
Lethal and Sub-Lethal Effects of Pyriproxyfen on Demography of Convergent Lady Beetle, Hippodamia convergens (Goeze) (Coccinellidae: Coleoptera)
To further develop integrated pest management (IPM) tactics against insect pests, demographic toxicology is considered important and efficient to evaluate the long-term effects of pesticides on biological control agents. In this study, lethal and sub-lethal effects of Pyriproxyfen (insect growth regulator) two concentrations of LC10 and LC30 were tested on second instar larvae of convergent lady beetle, Hippodamia convergens (Goeze) in order to evaluate the effect of insecticide on demographic parameters of the predator under laboratory conditions. The life table parameters were analysed statistically by using age-stage, two sex life table procedure. The results of this study show that developmental time for immature was prolonged in treated population (LC30 and LC10) rather than in control. Similarly, male and female longevity was also longer in the control group as compared to the treated population. Adult pre-oviposition period and fecundity were also greater in control as compared to the treated population. In addition, population parameters such as net reproductive rate (R0), intrinsic rate of increase (r) and finite rate of increase (λ) were also greater in control group rather than treated population. However, mean generation time (T) was greater in the treated group. The results revealed that pyriproxyfen, even at low concentrations, has potential to greatly affect the population growth of predatory lady beetle, therefore care should be taken when insect growth regulators are used within an IPM framework.
Diversity of Insect Pests of Paddy in Panhala Tehasil, Kolhapur, Maharashtra, India
Agriculture is the backbone of Indian economy and India is one of the world’s largest producers of Rice. Today, paddy crop is facing a severe problem of insect pests and is attacked by more than 100 species of insects, among those 20 species cause economic damage. Rice is the staple food of people of panhala tehasil, Kolhapur, Maharashtra, India. During June 2017 to September 2017 efforts were made to study the diversity of insect pests associated with the paddy crop in the study region. The collection and preservation of the specimens were done by following standard procedure and the identification was done with the help standard literature, taxonomic keys, and webography. In all, 6 species were recorded as pests of paddy in which order Lepidoptera was dominant with 2 species, while orders Diptera, Orthoptera, Hemiptera, and Coleoptera were represented by 1 species each respectively. The results of the present investigation will be helpful for formulating control strategies against these paddy pests.
Large-Scale Screening for Membrane Protein Interactions Involved in Platelet-Monocyte Interactions
Background: Beyond the classical roles in haemostasis and thrombosis, platelets are important in the initiation and development of various thrombo-inflammatory diseases. In atherosclerosis and deep vein thrombosis, for example, platelets bridge monocytes with endothelium and form heterotypic aggregates with monocytes in the circulation. This can alter monocyte phenotype by inducing their activation, stimulating adhesion and migration. These interactions involve cell surface receptor-ligand pairs on both cells. This list is likely incomplete as new interactions of importance to platelet biology are continuing to be discovered as illustrated by our discovery of PEAR-1 binding to FcεR1α. Results: We have developed a highly sensitive avidity-based assay to identify novel extracellular interactions among 126 recombinantly-expressed platelet cell surface and secreted proteins involved in platelet aggregation. In this study, we will use this method to identify novel platelet-monocyte interactions. We aim to identify ligands for orphan receptors and novel partners of well-known proteins. Identified interactions will be studied in preliminary functional assays to demonstrate relevance to the inflammatory processes supporting atherogenesis. Conclusions: Platelet-monocyte interactions are essential for the development of thromboinflammatory disease. Up until relatively recently, technologies only allow us to limit our studies on each individual protein interaction at a single time. These studies propose for the first time to study the cell surface platelet-monocyte interactions in a systematic large-scale approach using a reliable screening method we have developed. If successful, this will likely to identify previously unknown ligands for important receptors that will be investigated in details and also provide a list of novel interactions for the field. This should stimulate studies on developing alternative therapeutic strategies to treat vascular inflammatory disorders such as atherosclerosis, DVT and sepsis and other clinically important inflammatory conditions.
Helicoverpa armigera Hubner (Lepidoptera: Noctuidae) Susceptibility to Bacillus thuringiensis Crystal Toxins
Bacillus thuringiensis is a gram-positive spore-forming bacterium that belongs to the Bacillus cereus group of Bacilli and it produces ICP (insecticidal crystal protein) Cry toxins or Cysts toxins. Spores are produced as parasporal crystalline inclusions bodies (also known as endotoxins) at the onset of sporulation during the stationary growth phase. During vegetative growth that does not form crystals and is called vegetative insecticidal proteins (VIP) and secreted an insecticidal protein (SIP). Bacillus thuringiensis (Bt) is important for pest management either in the form of insecticides or through incorporated in the gene of the crop. Bioassays were conducted on the F2 generation of 1st instar larvae of H. armigera by the diet incorporation method to determine the susceptibility to Bt Cry toxins (Cry1Ac, Cry2Ab, Cry2A). The median lethal concentration (LC₅₀) of Cry1Ac, Cry2Ab, Cry2A ranged from 0.11 to 1.06 µg/ml and moult inhibitory concentration (MIC₅₀) of Cry1Ac, Cry2Ab, Cry2A ranged from 0.05 to 0.25 µg/ml. Cry1Ac was found most toxic to 1st instar larvae of H. armigera as compared to other Bt Cry toxins (Cry1Ac, Cry2Ab, Cry2A). The experimental results are important to policy-makers and technology providers to develop strategies for the exploitation of transgenic Bt cotton varieties as a component of integrated pest management.
A 7 Dimensional-Quantitative Structure-Activity Relationship Approach Combining Quantum Mechanics Based Grid and Solvation Models to Predict Hotspots and Kinetic Properties of Mutated Enzymes: An Enzyme Engineering Perspective
Enzymes are molecular machines used in various industries such as pharmaceuticals, cosmetics, food and animal feed, paper and leather processing, biofuel, and etc. Nevertheless, this has been possible only by the breath-taking efforts of the chemists and biologists to evolve/engineer these mysterious biomolecules to work the needful. Main agenda of this enzyme engineering project is to derive screening and selection tools to obtain focused libraries of enzyme variants with desired qualities. The methodologies for this research include the well-established directed evolution, rational redesign and relatively less established yet much faster and accurate insilico methods. This concept was initiated as a Receptor Rependent-4Dimensional Quantitative Structure Activity Relationship (RD-4D-QSAR) to predict kinetic properties of enzymes and extended here to study transaminase by a 7D QSAR approach. Induced-fit scenarios were explored using Quantum Mechanics/Molecular Mechanics (QM/MM) simulations which were then placed in a grid that stores interactions energies derived from QM parameters (QMgrid). In this study, the mutated enzymes were immersed completely inside the QMgrid and this was combined with solvation models to predict descriptors. After statistical screening of descriptors, QSAR models showed > 90% specificity and > 85% sensitivity towards the experimental activity. Mapping descriptors on the enzyme structure revealed hotspots important to enhance the enantioselectivity of the enzyme.
Trapping Efficiency of Highly Effective Slow Released Formulations of Biodegradable Waxes with Methyl Eugenol Against Bactrocera zonata
Experiment was carried out to evaluate the performance of highly effective Slow-Released Formulations (SRF) of Methyl eugenol with Lanolin wax, Candellila wax, Bee-wax, Carnauba wax and paraffin wax in the orchard of University of Agriculture Faisalabad, Pakistan against fruit flies. The waxes were mixed with methyl eugenol in 1:9 ratio. The results revealed that SRF of Candellila, Paraffin, Bees and Carnauba wax attracted 13.77, 11, 8.15 and 7.23 flies/day/trap which was 2.6, 2, 1.5 and 1.4 times higher than standard respectively and exhibited 41.42%, 32.05%, 20.98% and 12.87% attractive index respectively, proved moderately attractive slow-released formulation to B. zonata and was catagorized as Class-II slow-released formulation (AI = 11-50%). However, SRF of Lanolin wax trapped 1.81 flies/day/trap which was 3 times less than standard and exhibited -61.86% attractive index proved little or non attractive slow-released formulation and was categorized as Class-I slow-released formulation for B. zonata (AI < 11%).
Assesment of Trapping Efficiency of Slow Released Formulations of Methyl Euginol with Carnauba Wax against Bactrocera zonata
Present study was carried out to evaluate the performance of Slow-Released Formulations (SRF) of methyl eugenol with Carnauba wax in orchard of University of Agriculture Faisalabad, Pakistan against fruit flies. Carnauba wax was mixed with methyl eugenol in nine ratios (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20 and 90:10). The results revealed that SRFCN-9 trapped 35.3 flies/day/trap, exhibited an attractancy index (AI) of 50.35%, proved strongly attractive SRFCN for B. zonata and was categorized as Class-III slow-released formulation (Attractive Index > 50%). The SRFCN-1, SRFCN-2, SRFCN-3, SRFCN-4, SRFCN-5, SRFCN-6, SRFCN-7 and SRFCN-8 trapped 2.0, 5.3, 3.3, 4.0, 5.7, 12.0, 9.7 and 14.3 flies/day/trap respectively exhibited an attractancy index (AI) of -70.73%, -37.25%, -55.55%, -48.93%, -34.61%, 1.40%, -9.37% and 10.25% Attractive Index respectively, proved little or non attractive slow-released formulation and was categorized as Class-I slow-released formulation for B. zonata (Attractive Index < 11%). Results revealed that the Slow-Released Formulation containing 10% Carnauba wax with 90% methyl eugenol trapped maximum number of flies of over 30 days.
Assessment of Toxic Impact of Metals on Different Instars of Silkworm, Bombyx Mori
Larvae of silkworm (Bombyx mori) exhibit very high mortality when reared on mulberry leaves collected from mulberry orchards which get contaminated with metallic/nonmetallic compounds through either drift-deposition or chemigation. There is need to screen out such metallic compound for their toxicity at their various concentrations. The present study was carried out to assess toxicity of metals in different instars of silkworm. Aqueous solutions of nine heavy-metal based salts were prepared by dissolving 50, 100, 150, 200, 250, 300, 350 and 400 mg of each salt in one liter of water and were applied on the mulberry leaves by leaf-dip methods. The results reveal that mortality in 1st, 2nd, 3rd, 4th and 5th instar larvae caused by each heavy metal salts increased with an increase in their concentrations. The 1st instar larvae were found more susceptible to metal salts followed by 2nd, 3rd, 4th and 5th instar larvae of silkworm. Overall, Nickel chloride proved more toxic for all larval instar as it demonstrated approximately 40-99% mortality. On the basis of LC2 and larval mortality, the order of toxicity of heavy metals against all five larval instar was Nickel chloride (LC₂ = 1.9-13.9 mg/L; & 15.0±1.2-69.2±1.7% mortality) followed by Chromium nitrate (LC₂ = 3.3-14.8 mg/L; & 13.3±1.4-62.4±2.8% mortality), Cobalt nitrate (LC₂ = 4.3-30.9; &11.4±0.07-54.9±2.0% mortality), Lead acetate (LC₂ =8.8-53.3 mg/L; & 9.5±1.3-46.4±2.9% mortality), Aluminum sulfate (LC₂ = 15.5-76.6 mg/L; & 8.4±0.08-42.1±2.8% mortality), Barium sulfide (LC₂ = 20.9-105.9; & 7.7±1.1-39.2±2.5% mortality), Copper sulfate (LC2 = 28.5-12.4 mg/L; & 7.3±0.06-37.1±2.4% mortality), Manganese chloride (LC₂ = 29.9-136.9 mg/L; & 6.8±0.09-35.3±1.6% mortality) and Zinc nitrate (LC₂ = 36.3-15 mg/L; & 6.2±1.2-32.1±1.9% mortality). Zinc nitrate @ 50 and 100 mg/L, Barium sulfide @ 50 mg/L, Manganese chloride @ 50 and 100 mg/L and Copper sulfate @ 50 mg/L proved safe for 5th instar larvae as these interaction attributed no mortality. All the heavy metal salts at a concentration of 50 mg/L demonstrated less than 10% mortality.
Performance of Different Biodegradable Waxes Based Specialized Pheromone and Lure Application Technology-Male Anhelation Technique-Cue Lure Formulations in Bittergourd Field against Bactrocera cucurbitae
Melon fruit flies (Diptera: Tephritidae: Dacinae) are economically important pests of the cucurbits and are geographically distributed throughout the tropics and subtropics of the world. It causes heavy quantitative and qualitative losses in bitter gourd. The present experiment was carried out to evaluate the performance of different biodegradable waxes based SPLAT-MAT-CL (Specialized Pheromone and Lure Application Technology-Male Anhelation Technique- Cue Lure) formulations in bitter gourd field. Fourteen SPLAT-MAT emulsions/formulations were prepared by admixing different SPLAT matrices with toxicant (spinosad) and sex pheromone cuelure (attractant) in different proportionate percentage by weight. The results revealed that attraction and trapping of fruit flies of B. cucurbitae varied significantly for different SPLAT-MAT-CL formulations (p < 0.05). The maximum B. cucurbitae males were trapped in SPLAT-MAT-CL-7 (60 flies/trap/day) followed by SPLAT-MAT-CL-9 (40 flies/trap/day). The performance of all other formulations of SPLAT-MAT-CL was found in the order of SPLAT-MAT-CL-8 (30 flies/trap/day) > SPLAT-MAT-CL-3 (28 flies/trap/day) > SPLAT-MAT-CL-5 (25 flies/trap/day) > SPLAT-MAT-CL-4 (22 flies/trap/day) > SPLAT-MAT-CL-12 (20 flies/trap/day) SPLAT-MAT-CL-2 (19 flies/trap/day) > SPLAT-MAT-CL-14 (17 flies/trap/day) > SPLAT-MAT-CL-13 (15 flies/trap/day) > SPLAT-MAT-CL-11 (10 flies/trap/day) > SPLAT-MAT-CL-1 (8 flies/trap/day) > SPLAT-MAT-CL-10 (02 flies/trap/day). Overall, all the SPLAT-MAT-CL formulations, except SPLAT-MAT-CL-10, demonstrated higher density of captures of B. cucurbitae males as compared to standard (06 flies/trap/day). The results also demonstrate that SPLAT-MAT-CL-7, SPLAT-MAT-CL-9, SPLAT-MAT-CL-8, SPLAT-MAT-CL-3, SPLAT-MAT-CL-5, SPLAT-MAT-CL-4, SPLAT-MAT-CL-12, SPLAT-MAT-CL-2, SPLAT-MAT-CL-14, SPLAT-MAT-CL-13, SPLAT-MAT-CL-11 and SPLAT-MAT-CL-1 explained approximately 5, 4.6, 4.1, 3.6, 3.3, 3.1,2.8,2.5 and 1.6 times higher captures of B. cucurbitae males over standards. However, SPLAT-MAT-CL-10 demonstrated 3 times fewer captures of B. cucurbitae males over standards. In conclusion, SPLAT-MAT-CL-7, SPLAT-MAT-CL-9 can be exploited for the monitoring and trapping of B. cucurbitae in its IPM of program.
Assessment of Attractency of Bactrocera Zonata and Bactrocera dorsalis (Diptera:Tephritidae) to Different Biolure Phagostimulant-Mixtures
Fruit flies of Bactrocera genus cause heavy losses in fruits and vegetables globally and insecticide-application for their control creates issues of ecological backlash, environmental pollution, and food safety. There is need to explore alternatives and food-baits application is considered safe for the environment and effective for fruit fly management. Present experiment was carried out to assess the attractancy of five phagostimulant-Mixtures (PHS-Mix) prepared by mixing banana-squash, mulberry, protein-hydrolysate and molasses with some phagostimulant-lure sources including beef extract, fish extract, yeast, starch, rose oil, casein and cedar oil in five different ratios i.e., PHS-Mix-1 (1 part of all ingredients), PHS-Mix-2 (1 part of banana with 0.75 parts of all other ingredients), PHS-Mix-3 (1 part of banana with 0.5 parts of all other ingredients), PHS-Mix-4 (1 part of banana with 0.25 parts of all other ingredients) and PHS-Mix-5 (1 part of banana with 0.125 parts of all other ingredients). These were evaluated in comparison with a standard (GF-120). PHS-Mix-4 demonstrated 40.5±1.3-46.2±1.6% AI for satiated flies (class-II i.e., moderately attractive) and 59.5±2.0-68.6±3.0% AI for starved flies (class-III i.e., highly attractive) for both B. dorsalis and B. zonata in olfactometric study while the same exhibited 51.2±0.53% AI (class-III i.e., highly attractive) for B. zonata and 45.4±0.89% AI (class-II i.e., moderately attractive) for B. dorsalis in field study. PHS-Mix-1 proved non-attractive (class-I) and moderately attractive (class-II) phagostimulant in olfactometer and field studies, respectively. PHS-Mix-2 exhibited moderate attractiveness for starved lots in olfactometer and field-lot in field studies. PHS-Mix-5 proved non-attractive to starved and satiated lots of B. zonata and B. dorsalis females in olfactometer and field studies. Overall PHS-Mix-4 proved better phagostimulant-mixture followed by PHS-Mix-3 which was categorized as class-II (moderately attractive) phagostimulant for starved and satiated lots of female flies of both species in olfactometer and field studies; hence these can be exploited for fruit fly management.
Evaluation of Trapping Efficiency of Slow Released Formulations of Methyl Eugenol with Lanolin Wax against Bactrocera zonata
The study was carried out to evaluate the performance of Slow-Released Formulations (SRF) of Methyl eugenol with Lanolin wax in orchard of the University of Agriculture Faisalabad, Pakistan against fruit flies. Lanolin wax was mixed with methyl eugenol in nine ratios (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20 and 90:10). The results revealed that SRFₗₗ-7 trapped 42.1 flies /day/trap, exhibited an attractancy index (AI) of 51.71%, proved strongly attractive SRFₗₗ for B. zonata and was categorized as Class-III slow-released formulation (AI > 50%). The SRFₗₗ-2, SRFₗₗ-3, SRFₗₗ-4, SRFₗₗ-5, SRFₗₗ-6, SRFₗₗ-8 and SRFₗₗ-9 trapped 17.7, 27.9, 32.3, 23.8, 28.3, 37.8 and 19.9 flies /day/trap, exhibited an attractancy index (AI) of 20.54%, 41.02%, 26.00%, 34.15%, 43.50%, 49.86% and 46.07% AI respectively, proved moderately attractive slow-released formulations for B. zonata and were categorized as Class-II slow-released formulations (AI = 11-50%). However, SRFₗₗ-1 trapped 14.8 flies /day/trap, exhibited 0.71% AI proved little or nonattractive slow-released formulation and was categorized as Class-I slow-released formulation for B. zonata (AI < 11%).
Taxonomic and Faunistic Data on the Genus Triaspis Haliday, 1835 (Hymenoptera: Braconidae: Brachistinae) from Turkey
Brachistinae F&ouml;erster, 1862 is a subfamily of the family Braconidae (order Hymenoptera) with about 410 species distributed all around the world. Brachistinae includes the genera, Eubazus Nees von Esenbeck 1814, Foersteria Sz&eacute;pligeti 1896, Chelostes van Achterberg 1990, Triaspis Haliday 1835 and Schizoprymnus F&ouml;rster 1862. Members of the subfamily live as parasitoids on the families Curculionidae and Apionidae (Coleoptera), which also include very important agricultural pests.&nbsp; In generally, members of the genus Triaspis are poorly known biologically. The genus is represented by 37 species in the West Palearctic region and 118 species worldwide. Adult specimens of Triaspis were collected from as wide a range of habitats as possible at different altitudes in different parts of Turkey between 1982 and 2010. Samples collected from short plants using standard insect sweeping nets were transferred into tubes containing 70% ethanol and labelled following their preparations according to museum techniques. Seven Triaspis species have been reported from Turkey in this study. Five of these species are new to the fauna of Turkey.
An Insight into the Conformational Dynamics of Glycan through Molecular Dynamics Simulation
Glycan of glycolipids and glycoproteins is playing a significant role in living systems particularly in molecular recognition processes. Molecular recognition processes are attributed to their occurrence on the surface of the cell, sequential arrangement and type of sugar molecules present in the oligosaccharide structure and glyosidic linkage diversity (glycoinformatics) and conformational diversity (glycoconformatics). Molecular Dynamics Simulation study is a theoretical-cum-computational tool successfully utilized to establish glycoconformatics of glycan. The study on various oligosaccharides of glycan clearly indicates that oligosaccharides do exist in multiple conformational states and these conformational states arise due to the flexibility associated with a glycosidic torsional angle (φ,ψ) . As an example: a single disaccharide structure NeuNacα(2-3) Gal exists in three different conformational states due to the differences in the preferential value of glycosidic torsional angles (φ,ψ). Hence establishing three dimensional structural and conformational models for glycan (cartesian coordinates of every individual atoms of an oligosaccharide structure in a preferred conformation) is quite crucial to understand various molecular recognition processes such as glycan-toxin interaction and glycan-virus interaction. The gycoconformatics models obtained for various glycan through Molecular Dynamics Simulation stored in our 3DSDSCAR (3DSDSCAR.ORG) a public domain database and its utility value in understanding the molecular recognition processes and in drug design venture will be discussed.
An Insight into the Conformational Dynamics of Glycan through Molecular Dynamics Simulation
Glycan of glycolipids and glycoproteins is playing a significant role in living systems particularly in molecular recognition processes. Molecular recognition processes are attributed to their occurrence on the surface of the cell, sequential arrangement and type of sugar molecules present in the oligosaccharide structure and glyosidic linkage diversity (glycoinformatics) and conformational diversity (glycoconformatics). Molecular Dynamics Simulation study is a theoretical-cum-computational tool successfully utilized to establish glycoconformatics of glycan. The study on various oligosaccharides of glycan clearly indicates that oligosaccharides do exist in multiple conformational states and these conformational states arise due to the flexibility associated with a glycosidic torsional angle (φ,ψ) . As an example: a single disaccharide structure NeuNacα(2-3) Gal exists in three different conformational states due to the differences in the preferential value of glycosidic torsional angles (φ,ψ). Hence establishing three dimensional structural and conformational models for glycan (cartesian coordinates of every individual atoms of an oligosaccharide structure in a preferred conformation) is quite crucial to understand various molecular recognition processes such as glycan-toxin interaction and glycan-virus interaction. The gycoconformatics models obtained for various glycan through Molecular Dynamics Simulation stored in our 3DSDSCAR (3DSDSCAR.ORG) a public domain database and its utility value in understanding the molecular recognition processes and in drug design venture will be discussed.
Removal of Maxilon Red Dye by Adsorption and Photocatalysis: Optimum Conditions, Equilibrium, and Kinetic Studies
The present work has for main objective the elimination of the textile dye Maxilon Red (MR) by two processes, adsorption on activated clay followed by photocatalysis in presence of ZnO as a photocatalyst. The influence of the physical parameters like the initial pH, adsorbent dose of the activated clay, the MR concentration and temperature has been studied. The best adsorption yield occurs at neutral pH ~ 7 within 60 min with an uptake percentage of 97% for a concentration of 25 mg L⁻¹ and a dose of 0.5 g L⁻¹. The adsorption data were suitably fitted by the Langmuir model with a maximum capacity of 176 mg g⁻¹. The MR adsorption is well described by the pseudo second order kinetic. The second part of this work was dedicated to the photocatalytic degradation onto ZnO under solar irradiation of the residual MR concentration, remained after adsorption. The effect of ZnO dose and MR concentration has also been investigated. The parametric study showed that the elimination is very effective by this process, based essentially on the in situ generation of free radicals *OH which are non-selective and very reactive. The photodegradation process follows a first order kinetic model according to the Langmuir-Hinshelwood model.
A Faunistic Study of Tetranychid and Phytoseiid Mites Associated with Diverse Crops From Samsun, Turkey
This research was implemented from March to September to reveal tetranychid and phytoseiid mites on different field crops in Samsun province, Turkey. In consequence of microscope slide-mounting of mite samples in Hoyer’s medium, a total of six species belonging to Tetranychidae and fourteen species belonging to Phytoseiidae were found. Tetranychus urticae Koch, Tetranychus turkestani Ugarov and Nikolski, Tetranychus viennensis Zacher, Panonychus ulmi (Koch), Panonychus citri (Mc Gregor) and Bryobia rubrioculus (Scheuten) were detected as phytophaous mites. Euseius finlandicus (Oudemans), Kampimodromus aberrans (Oudemans), Amblyseius agrestris (Karg), Amblyseius andersoni (Chant), Amblyseius bicaudus Wainstein, Amblyseius zwölferi (Dosse), Amblyseius barkeri (Hughes), Paraseilus soleiger (Ribaga), Anthoseius recki (Wainstein), Phytoseius finitimus Ribaga, Typhlodromus pyri Scheuten, Typhloctonus tiliarum Oudemans, Phytoseiulus macropilis (Banks) and Phytoseiulus persimilis Athias-Henriot were identified to be predatory mites in Phytoseiidae. Among the phytoseiid species Kampimodromus aberrans, Amblyseius andersoni, Anthoseius recki, Phytoseius finitimus, Phytoseiulus persimilis and Phytoseiulus macropilis were widespread. Relationship between tetranychid and phytoseiid mites on different crops that can make considerable contribution to biological control in integrated pest management (IPM) programs is also reported.
Nucleotide Diversity and Bacterial Endosymbionts of the Black Cherry Aphid Myzus cerasi (Fabricus, 1775) (Hemiptera: Aphididae) from Turkey
Sequences of mitochondrial cytochrome oxidase I (COI) gene of twenty-five Turkish and one Greek Myzus cerasi (Fabricus) (Hemiptera: Aphididae) in populations were collected from Prunus avium and Prunus cerasus. The partial coding region of COI studied is 605 bp for all the populations, from which 565 nucleotides were conserved, 40 were variable, 37 were singleton, and 3 sites were parsimony-informative. Four haplotypes were identified based on nucleotide substitutions, and the mean of intraspecific divergence was calculated to be 0.3%. Phylogenetic trees were constructed using Maximum Likelihood, Minimum Evolution, Neighbor-joining, and Unweighed Pair Group Method of Arithmetic Averages (UPGMA) and Myzus persicae (Sulzer) and Myzus borealis Ossiannilson were included as outgroups. The population of M. cerasi from Isparta diverged from the rest of the groups and formed a clade (Haplotype B) with Myzus borealis. The rest of the haplotype diversity includes Haplotype A and Haplotype C with individuals characterized as Myzus cerasi pruniavium and Haplotype D with Myzus cerasi cerasi. M. cerasi diverge into two subspecies and it must be reevaluated whether this pest is monophagous or oligophagous in terms of plant type dependence. The obligated endosymbiont Buchnera aphidicola was also found during this research, but no facultative symbionts could be found. It is expected further studies will be required for a complete barcoding and diversity of bacterial endosymbionts present.
Systematics of Water Lilies (Genus nymphaea L.) Using 18S rDNA Sequences
The waterlilies (Nymphaea L.) is the largest genus of Nymphaeaceae. This family is composed of six genera (Nuphar, Ondinea, Euryale, Victoria, Barclaya, Nymphaea). Its members are nearly worldwide in tropical and temperate regions. The classification of some species in Nymphaea has complicated in high variation of petal and leaves. Therefore, the phylogenetic relationships based on 18S rDNA were constructed to delimit this genus. DNA of fifty-two specimens belonging to waterlily family was extracted using modified conventional method containing cetyltrimethyl ammonium bromide (CTAB). The results showed that the amplified fragment is about 1600 basepairs in size. After analysis, the aligned sequences presented 9.36% for variable characters comprising 2.66% of parsimonious informative sites and 6.70% of singleton sites. Moreover, there are 6 regions of 1-2 base(s) for insertion/deletion. The phylogenetic trees based on maximum parsimony and maximum likelihood with high bootstrap support indicated that genus Nymphaea was a paraphyletic group because of Ondinea, Victoria and Euryale disruption. Within genus Nymphaea, subgenus Nymphaea is a basal lineage group which cooperated with Euryale and Victoria. The other four subgenera, namely Lotos, Hydrocallis, Brachyceras and Anecphya were included the same large clade which Ondinea was placed within Anecphya clade due to geographical sharing.
Impact of Different Rearing Diets on the Performance of Adult Mealworms Tenebrio molitor
Production of insects for human and animal consumption is an increasingly important activity in Canada. Protein production is more efficient and less harmful to the environment using insect rearing compared to the impact of traditional livestock, poultry and fish farms. Insects are rich in essential amino acids, essential fatty acids and trace elements. Thus, insect-based products could be used as a food supplement for livestock and domestic animals and may even find their way into the diets of high performing athletes or fine dining. Nevertheless, several parameters remain to be determined to ensure efficient and profitable production that meet the potential of these sectors. This project proposes to improve the production processes, rearing diets and processing methods for three species with valuable gastronomic and nutritional potential: the common mealworms (Tenebrio molitor), the small mealworm (Alphitobius diaperinus), and the giant mealworm (Zophobas morio). The general objective of the project is to acquire specific knowledge for mass rearing of insects dedicated to animal and human consumption in order to respond to current market opportunities and meet a growing demand for these products. Mass rearing of the three species of mealworm was produced to provide the individuals needed for the experiments. Mealworms eat flour from different cereals (e.g. wheat, barley, buckwheat). These cereals vary in their composition (protein, carbohydrates, fiber, vitamins, antioxidant, etc.), but also in their purchase cost. Seven different diets were compared to optimize the yield of the rearing. Diets were composed of cereal flour (e.g. wheat, barley) and were either mixed or left alone. Female fecundity, larvae mortality and growing curves were observed. Some flour diets have positive effects on female fecundity and larvae performance while each mealworm was found to have specific diet requirements. Trade-offs between mealworm performance and costs need to be considered. Experiments on the effect of flour composition on several parameters related to performance and nutritional and gastronomic value led to the identification of a more appropriate diet for each mealworm.
Rhizosphere Microbiome Involvement in the Natural Suppression of Soybean Cyst Nematode in Disease Suppressive Soil
The rhizosphere microbiome elucidate multiple functioning in the soil suppressiveness against plant pathogens. Soybean rhizosphere microbial communities may involve in the natural suppression of soybean cyst nematode (SCN) populations in disease suppressive soils. To explore these ecological mechanisms of microbes, a long term monoculture suppressive soil were taken into account for further investigation to test the disease suppressive ability by using different treatments. The designed treatments are as, i) suppressive soil (S), ii) conducive soil (C), iii) conducive soil mixed with 10% (w/w) suppressive soil (CS), iv) suppressive soil treated at 80°C for 1 hr (S80), and v) suppressive soil treated with formalin (SF). By using an ultra-high-throughput sequencing approach, we identified the key bacterial and fungal taxa involved in SCN suppression. The Phylum-level investigation of bacteria revealed that Actinobacteria, Bacteroidetes, and Proteobacteria in the rhizosphere soil of soybean seedlings were more abundant in the suppressive soil than in the conducive soil. The phylum-level analysis of fungi in rhizosphere soil indicated that relative abundance of Ascomycota was higher in suppressive soil than in the conducive soil, where Basidiomycota was more abundant. Transferring suppressive soil to conducive soil increased the population of Ascomycota in the conducive soil by lowering the populations of Basidiomycota. The genera, such as, Pochonia, Purpureocillium, Fusarium, Stachybotrys that have been well documented as bio-control agents of plant nematodes were far more in the disease suppressive soils. Our results suggested that the plants engage a subset of functional microbial groups in the rhizosphere for initial defense upon nematode attack and protect the plant roots later on by nematodes to response for suppression of SCN in disease-suppressive soils.
The Advantages of Using DNA-Barcoding for Determining the Fraud in Seafood
Although seafood is an important part of human diet and categorized highly traded food industry internationally, it is remain overlooked generally in the global food security aspect. Food product authentication is the main interest in the aim of both avoids commercial fraud and to consider the risks that might be harmful to human health safety. In recent years, with increasing consumer demand for regarding food content and it's transparency, there are some instrumental analyses emerging for determining food fraud depend on some analytical methodologies such as proteomic and metabolomics. While, fish and seafood consumed as fresh previously, within advanced technology, processed or packaged seafood consumption have increased. After processing or packaging seafood, morphological identification is impossible when some of the external features have been removed. The main fish and seafood quality-related issues are the authentications of seafood contents such as mislabelling products which may be contaminated and replacement partly or completely, by lower quality or cheaper ones. For all mentioned reasons, truthful consistent and easily applicable analytical methods are needed for assurance the correct labelling and verifying of seafood products. DNA-barcoding methods become popular robust that used in taxonomic research for endangered or cryptic species in recent years; they are used for determining food traceability also. In this review, when comparing the other proteomic and metabolic analysis, DNA-based methods are allowing a chance to identification all type of food even as raw, spiced and processed products. This privilege caused by DNA is a comparatively stable molecule than protein and other molecules. Furthermore showing variations in sequence based on different species and founding in all organisms, make DNA-based analysis more preferable. This review was performed to clarify the main advantages of using DNA-barcoding for determining seafood fraud among other techniques.
Transcriptional Response of Honey Bee to Differential Nutritional Status and Nosema Infection
Bees are confronting several environmental challenges, including the intermingled effects of malnutrition and disease. Intuitively, pollen is the healthiest nutritional choice; however, commercial substitutes, such as BeePro and MegaBee, are widely used. Herein we examined how feeding natural and artificial diets shapes transcription in the abdomen of the honey bee, and how transcription shifts in combination with Nosema parasitism. Gene ontology enrichment revealed that, compared with poor diet (carbohydrates (C)), bees fed pollen (P > C), BeePro (B > C), and MegaBee (M > C) showed a broad upregulation of metabolic processes, especially lipids; however, pollen feeding promoted more functions and superior proteolysis. The superiority of the pollen diet was also evident through the remarkable overexpression of vitellogenin in bees fed pollen instead of MegaBee or BeePro. Upregulation of bioprocesses under carbohydrates feeding compared to pollen (C > P) provided a clear poor nutritional status, uncovering stark expression changes that were slight or absent relatively to BeePro (C > B) or MegaBee (C > M). Poor diet feeding (C > P) induced starvation response genes and hippo signaling pathway, while it repressed growth through different mechanisms. Carbohydrate feeding (C > P) also elicited ‘adult behavior’, and developmental processes suggesting transition to foraging. Finally, it altered the ‘circadian rhythm’, reflecting the role of this mechanism in the adaptation to nutritional stress in mammals. Nosema-infected bees fed pollen compared to carbohydrates (PN > CN) upheld certain bioprocesses of uninfected bees (P > C). Poor nutritional status was more apparent against pollen (CN > PN) than BeePro (CN > BN) or MegaBee (CN > MN). Nosema accentuated the effects of malnutrition since more starvation-response genes and stress response mechanisms were upregulated in CN > PN compared to C > P. The bioprocess ‘Macromolecular complex assembly’ was also enriched in CN > PN, and involved genes associated with human HIV and/or influenza, thus providing potential candidates for bee-Nosema interactions. Finally, the enzyme Duox emerged as essential for guts defense in bees, similarly to Drosophila. These results provide evidence of the superior nutritional status of bees fed pollen instead of artificial substitutes in terms of overall health, even in the presence of a pathogen.