Search results for: bioassay

Authors: Lauretta Ighedo, Bukola Okunade, Monisade Okunade

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In order to determine the effect of spent oil on the growth rates of indigenous phytoplankton in an aquaculture pond, a study was carried out on varying concentrations of samples using the bioassay procedure for a period of 14 days. Four divisions Cyanophyta, Chlorophyta, Euglenophyta and Bacillariophyta were observed in the water samples collected from the Aquaculture pond. The growth response was measured using a microprocessor photocolorimeter at optical density of 680nm. A general assessment of spent oil contaminated samples showed either a sharp rise or fall in growth rate from day 0 to day 2 followed by increased growth response for most higher concentration of pollutants up to Day 8, then fluctuations in the growth response pattern for the other days. There was no marked significant difference in the growth response of phytoplankton in the spent oil impacted water samples. The lowest and highest phytoplankton abundance was recorded in 10/90ml and 2.5/97.5ml spent oil impacted water sample respectively. Oscillatoria limosa, Chlorella sp., Microcystis aeruginosa, Nitzschia sp. and Navicula sp. showed high tolerance to oil pollution and these species used as bioindicators of an organic polluted environment increased abundantly and can therefore be employed in the cleanup and bioremediation process of an oil polluted freshwater body.

Keywords: phytoplankton, pollution, species abundance, environmental characteristics

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Authors: Muhammad Tariq, Bushra Siddique, Muhammad Naeem, Asim Gulzar

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The chemical ecology of natural enemies can play a pivotal role in any Integrated Pest Management (IPM) program. Different chemical cues help to correspond in the diversity of associations between prey and host plant species. Coccinellaseptempunctata and Diaeretiellarapae have the abilities to explore several chemical cues released by plants under herbivore attack that may enhance their efficiency of foraging. In this study, the behavioral responses of Coccinellaseptempunctata and Diaeretiellarapae were examined under the application of two semiochemicals and a plant extract and their combinations using four-arm olfactometer. The bioassay was consists of a pairwise treatment comparison. Data pertaining to the preference of C. septempunctata and D. rapae after treatment application were recorded and analyzed statistically. The mean number of entries and time spent of Coccinellaseptempunctata and D. rapaewere greater in arms treated with E-β-Farnesene. However, the efficacy of E-β-Farnesene was enhanced when combined with β-pinene. Thus, the mean number of entries and time spent of C. septempunctata and D. rapaewere highest in arms treated with the combination of E-β-Farnesene x β-pinene as compared with other treatments. The current work has demonstrated that the insect-derived semiochemicals may enhance the efficacy of natural enemies when applied in combination.

Keywords: olfectometer, parasitoid, predator, preference

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Authors: Karim Saeidi

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One of the most important pests in the warehouses is the flour beetle, Tribolium confusum Jacquelin du Val (Col.: Tenebrionidae). Regarding the high degree of damage of stored product pests and dangerous effects of the chemical control using plant extracts and their components are some of the best approaches to control these pests. Antifeedant activity of plant extracts from Lavandula angustifolia Mill and Zataria multiflora Boiss using hydro-distillation were tested against the flour beetle, Tribolium confusum Jacquelin du Val. The nutritional indices: relative growth rate (RGR), relative consumption rate (RCR), the efficiency of conversion of ingested food (ECI), and feeding deterrence index (FDI) were measured for adult insects. Treatments were evaluated using a flour disk bioassay in the dark; at 25±1ᵒC and 60±5% R. H. Concentrations of 0, 0.1, 0.5, 0.75, 1, 1.5, and 2 μl/disk were prepared from each essential oil. After 72 h, nutritional indices were calculated. L. angustifolia oils were more effective than Z. multiflora oils by significantly decreasing the RGR, RCR, and ECI. Feeding deterrence index (FDI) of L. angustifolia essential oil was increased significantly as essential oil concentration increased. The essential oil of L. angustifolia was more effective on FDI than Z. multiflora in some concentration.

Keywords: essential oil, nutritional indices, Tribolium confusum

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Authors: Yusuf Ali Abdulle, Azhar Uddin Keerio

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Background: Protein elicitors play a key role in signaling or displaying plant defense mechanisms and emerging as vital tools for bio-control of insects. This study was aimed at the characterization of the novel protein elicitor isolated from entomopathogenic fungi Lecanicillium lecanii (V3) strain and its activity against Whitefly, Bemisia tabaci in cotton. The sequence of purified elicitor protein showed 100% similarity with hypothetical protein LEL_00878 [Cordyceps confragosa RCEF 1005], GenBank no (OAA81333.1). This novel protein elicitor has 253 amino acid residues and 762bp with a molecular mass of 29 kDa. The protein recombinant was expressed in Escherichia coli using pET‐28a (+) plasmid. Effects of purified novel protein elicitor on Bemisia tabaci were determined at three concentrations of protein (i.e., 58.32, 41.22, 35.41 μg mL⁻¹) on cotton plants and were exposed to newly molted adult B.tabaci. Bioassay results showed a significant effect of the exogenous application of novel protein elicitor on B. tabaci in cotton. In addition, the gene expression analysis found a significant up-regulation of the major genes associated with salicylic acid (SA) and jasmonic acid (JA) linked plant defense pathways in elicitor protein-treated plants. Our results suggested the potential application of a novel protein elicitor derived from Lecanicillium lecanii as a future bio-intensive controlling approach against the whitefly, Bemisia tabaci.

Keywords: resistance, Lecanicillium lecanii, secondary metabolites, whitefly

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Authors: Harshani Uggallage, Kapila D. Dissanayaka

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A Sulforhodamine-B assay-guided fractionation on Nigella sativa seeds was conducted to determine the presence of cytotoxic compounds against human hepatoma (HepG2) cells. Initially, a freeze-dried sample of Nigella sativa seeds was sequentially extracted into solvents of increasing polarities. Crude extracts from the sequential extraction of Nigella sativa seeds in chloroform and ethyl acetate showed the highest cytotoxicity. The combined mixture of these two extracts was subjected to bioassay guided fractionation using a modified Kupchan method of partitioning, followed by Sephadex® LH-20 chromatography. This chromatographic separation process resulted in a column fraction with a convincing IC50 (half-maximal inhibitory concentration) value of 13.07µg/ml, which is considerable for developing therapeutic drug leads against human hepatoma. Reversed phase High-Performance Liquid Chromatography (HPLC) was finally conducted for the same column fraction, and the result indicates the presence of one or several main cytotoxic compounds against human HepG2 cells.

Keywords: cytotoxic compounds, half-maximal inhibitory concentration, high-performance liquid chromatography, human HepG2 cells, nigella sativa seeds, Sulforhodamine-B assay

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Authors: Tayyaba Bibi, Jenny Ronan, Robert Hernan, Kathleen O’Rourke, Brendan McHugh, Evin McGovern, Michelle Giltrap, Gordon Chambers, James Wilson

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Sediment contamination is a major source of ecosystem stress and has received significant attention from the scientific community. Both the Water Framework Directive (WFD) and Marine Strategy Framework Directive (MSFD) require a robust set of tools for biological and chemical monitoring. For the MSFD in particular, causal links between contaminant and effects need to be assessed. Appropriate assessment tools are required in order to make an accurate evaluation. In this study, a range of recommended sediment bioassays and chemical measurements are assessed in a number of potentially impacted and lowly impacted locations around Ireland. Previously, assessment indices have been developed on individual compartments, i.e. contaminant levels or biomarker/bioassay responses. A number of assessment indices are applied to chemical and ecotoxicological data from the Seachange project (Project code) and compared including the metal pollution index (MPI), pollution load index (PLI) and Chapman index for chemistry as well as integrated biomarker response (IBR). The benefits and drawbacks of the use of indices and aggregation techniques are discussed. In addition to this, modelling of raw data is investigated to analyse links between contaminant and effects.

Keywords: bioassays, contamination indices, ecotoxicity, marine environment, sediments

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Authors: Zunnu Raen Akhtar

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Cotton Leaf Curl Disease (CLCuD) is from Gemini virus and is transmitted through whiteflies in cotton. Transgenic cotton containing Antisense Coat Protein (ACP) has been found to show better results against CLCuD in cotton. In current research, Antisense Coat Protein was inserted in cotton plants to observe resistance developed in the cotton plants against CLCuD. T1 generation of plants were observed for its expression in plants. Tests were carried out to observe the expression of Antisense Coat Protein using Polymerase Chain Reaction (PCR) technique and by southern blotting. Whiteflies showing positive Cotton Leaf Curl Virus (CLCV) were reared and released in bioassay on ACP expressing cotton plants under laboratory as well as confined semi-field conditions. Results confirmed the expression of AC protein in PCR and southern blotting. Further laboratory results showed that cotton plants expressing AC protein showed rare incidence of CLCuD infection as compared to control. In the confined semi-field, similar results were observed in AC protein expressing cotton as compared to control. These results explicitly show that ACP can help to tackle the CLCuD issue in the future and further studies on biochemical processes involved in these plants and effects of ACP induction on non-target organisms should also be studied for eco-system.

Keywords: cotton, white flies, antisense coat protein, CLCV

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Authors: Hadura Abu Hasan, Zairi Jaal, P. J. McCall

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Dengue is a serious problem in Malaysia, particularly in high-density urban communities with lower socio-economic levels. This study evaluated the susceptibility of local populations of Aedesaegypti (Linnaeus), Aedesalbopictus (Skuse) and Culexquinquefasciatus (Say) from the traditional community of BaganDalam, Penang, Malaysia to lambdacyhalothrin and pirimiphos-methyl using standard World Health Organization (WHO) adult bioassay test. Unfed female mosquitoes aged 3-5 days were exposed to WHO recommended dosages of insecticides over fixed time periods with results presented as knock-down time (KT50) for each strain.The insecticide susceptible VCRU laboratory strain was usedas control. All three specieswere highly resistant to lambda-cyhalothrin with less than 10% mortality at 24 hours after treatment. In contrast, Ae.aegypti and Ae. albopictus were susceptible to pirimiphos-methyl, showing 100% mortality recorded 24 hoursafter treatment. Cx. quinquefasciatuswasclassed as ‘suspected resistant’ to pirimiphos-methyl as mortality recorded 24 hours after treatment was 94-96%. The results indicate that organophosphates such as pirimiphos-methyl might be used as alternative to pyrethroid for dengue vector control in this dengue-prone area.

Keywords: vector control, aedes aegypti, aedes albopictus, dengue, culex quinquefasciatus, residuals insecticides, pyrethroid, organophosphate, resistant, mosquito

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Authors: Guru Prasad Satsangi, Shiv Shankar Gautam

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Allelopathy is a potential area of research for sustainable agriculture. It is environmentally safe, can conserve the available resources, and also may mitigate the problems raised by synthetic chemicals. The allelo-chemicals are secondary metabolites produced by plants, which are the byproducts of the primary metabolic process. These allelo-chemicals may be stimulatory, inhibitory, or may have no effect on the growth of the other plants. It has been observed in the present study that foliar extracts of Leucaena leucocephala showed an inhibitory effect on the germination of the test crop maize. The results revealed that at different concentrations of Leucaena leucocephala foliar extract, caused a significant inhibition in germination and growth behavior of Zea mays L. seedlings. Minimum germination and growth occurred in 100 % concentration, and an increase in extract concentrations result in a decrease in the germination. Bioassay also depicted that this inhibitory effect was proportional to the concentration of the extract as the higher concentration having a lesser stimulatory effect or vice versa. The phytochemical analysis of the secondary metabolites from foliar extracts of Leucaena leucocephala L. showed the presence of tannins, saponins, phenols, alkaloids, and flavanoids. Among various extracts, the presence of methanol extract was found in a significant amount of phytochemicals, followed by the aqueous and ethanol extracts. Leaves showed a significantly higher amount of the allelochemicals.

Keywords: allelopathic effect, germination /growth behavior , foliar extracts, Leucaena leucceophala , Zea mays L.

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Authors: Jarel Elgin Tolentino, Arby Denise Nera, Mary Rose Roco, Angela Vianca Aspa, Nikko Beltran, Else Dapat

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Drug resistance by cells has been the problem in the medical field for decades now. The use of medicinal plants as a source of creating powerful drugs has been nowadays recognized worldwide to treat such resistant diseases. In the present study, the potential for Diospyros philippinensis A. DC. to inhibit growth of both bacteria and cancer cell line was conducted. The leaf crude extracts were screened for the presence of phytochemicals and examined for potential bioactivities by employing several assays like Kirby-Bauer disc diffusion method, DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) assay and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) tetrazolium assay for the antibacterial, antioxidant and cytotoxic activities of the extract, respectively. Phytochemical test results of the extracts revealed the presence of alkaloids, flavonoids, saponins, phenols, quinones, cardiac glycosides, phlobatannins, carbohydrate, cardenolides and proteins. The leaf extracts were found to exhibit antibacterial activity against gram-positive bacteria, high antioxidant activity (99.22% ± 0.005) but did not show any sign of cytotoxicity towards HCT116 (ATCC CCL-247). The study therefore concludes that D. philippinensis A. DC. leaf extract can be a source of antibacterial and chemopreventive agents. This claim may be used as basis for future investigation.

Keywords: bioassay, medicinal plants, plant crude extracts, phytochemical screening

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Authors: Asim Abbasi, Muhammad Sufyan

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The present experiment was carried out to examine the effects of silicon dioxide on tritrophic interaction of cotton, whitefly, and the predator Chrysoperla carnea. Population of whitefly was maintained on silicon treated and non-treated cotton for two generations in greenhouse net cages exposed to outside temperature and luminosity. The cotton was treated with silicon dioxide twice after 15 days intervals with 200 ppm concentration. A stock rearing of the natural predator was developed in the laboratory conditions. In the bioassay eggs of the predator all at the same age were individualized in glass petri plates that will be pierced with a pin to allow aeration and maintained in an incubator at 28 ± 2°C, 70 ± 10% relative humidity and 12h photo phase. Population of whitefly stayed on silicon treated, and non-treated cotton were offered to newly hatched chrysopid larvae until the end of the larval stage, assuring a permanent supply. Feeding preference of C. carnea along with longevity, survival of each instar larvae, pupation, adult emergence, and fecundity was checked. The results revealed that there was no significant difference in the feeding preference of C. carnea among both treatments. Durations of 1st and 2nd larval instar were also at par in both treatments. However overall longevity and adult emergence were a bit lower in silicon treated whitefly treatment. This may be due to the fact that silicon reduces the nutritional quality of host because of reduced whitefly feeding on silicon treated cotton. No significant difference in 1st and 2nd larval instars and then increased larval duration in later instars suggested that the effect of silicon treated host should be checked on more than 1 generation of C. carnea to get better findings.

Keywords: Chrysoperla carnea, silicon, tritrophic, whitefly

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Authors: Zahid Rehman, TorOve Leiknes

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Quorum sensing (QS) is the process by which bacteria communicate with each other through small signaling molecules, such as N-acylhomoserine lactones (AHLs). Also, certain bacteria have the ability to degrade AHL molecules by a process referred to as quorum quenching (QQ); therefore, QQ can be used to control bacterial infections and biofilm formation. In this study, we aimed to identify new species of bacteria with QQ activities. To achieve this, sediments from Red Sea were collected either in the close vicinity of Sea grass or from area with no vegetation. From these samples, we isolated 72 bacterial strains and tested their ability to degrade/inactivate AHL molecules. Chromobacterium violaceum based bioassay was used in initial screening of isolates for QQ activity. The QQ activity of the positive isolates was further confirmed and quantified by employing liquid chromatography and mass spectrometry. These analyses showed that isolated bacterial strain could degrade AHL molecules with different acyl chain length and modifications. Sequencing of 16S-rRNA genes of positive isolates revealed that they belong to three different genera. Specifically, two isolates belong to genus Erythrobacter, four to Labrenzia and one isolate belongs to Bacterioplanes. Time course experiment showed that isolate belonging to genus Erythrobacter could degrade AHLs faster than other isolates. Furthermore, these isolates were tested for their ability to inhibit formation of biofilm and degradation of 3OXO-C12 AHLs produced by P. aeruginosa PAO1. Our results showed that isolate VG12 is better at controlling biofilm formation. This aligns with the ability of VG12 to cause at least 10-fold reduction in the amount of different AHLs tested.

Keywords: quorum sensing, biofilm, quorum quenching, anti-biofouling

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Authors: Arunporn Itharat, Kamonmas Srikwan, Srisopa Ruangnoo, Pakakrong Thongdeeying

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Background: The rhizomes of Smilax glabra (SG) has long been used in Traditional Chinese and Thai herbal medicine to treat a variety of infectious diseases and immunological disorders. Objective: To investigate the in vitro anti-allergic activities of crude extracts and pure isolated flavonoid compounds from SG by determination of inhibitory effects on antigen-induced release of β-hexosaminidase from RBL-2H3 cells. Methods: The in vitro inhibitory effects of crude aqueous and organic extracts on beta-hexosaminidase release in RBL-2H3 cells were evaluated as an in vitro indication of possible anti-allergic activity in vivo. Bioassay-guided fractionation of extracts was used to isolate flavonoid compounds from the ethanolic extracts. Results: The 95% and 50% ethanolic extracts of SG showed remarkably high anti-allergic activity, with IC50 values of 5.74 ± 2.44 and 23.54 ± 4.75 μg/ml, much higher activity than that for Ketotifen (IC50 58.90 μM). The water extract had negligible activity (IC50 > 100 μg/ml). The two isolated flavonols, Engeletin and Astilbin, showed weak anti-allergic activity, IC50 values 97.46 ± 2.04 and > 100 μg/ml, respectively. Conclusions: The 95% and 50% ethanolic extracts of SG showed strong anti-allergic activity but two flavonol constituents did not show any significant anti-allergic activity. These findings suggest that a combination of effects of various phytochemicals in crude extracts used in traditional medicine are responsible for the purported anti-allergic activity of SG herbal preparations. The plethora of constituents in crude extracts, as yet unidentified, are likely to be acting synergistically to account for the strong observed anti-allergic in vitro activity.

Keywords: Smilax glabra, anti-allergic activity, RBL-2H3 cells, flavonoid compounds

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Authors: U. Pangnakorn, S. Kanlaya

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The efficiency of wood vinegar mixed with each individual of three plants extract such as: citronella grass (Cymbopogon nardus), neem seed (Azadirachta indica A. Juss), and yam bean seed (Pachyrhizus erosus Urb.) were tested against the second instar larvae of housefly (Musca domestica L.). Steam distillation was used for extraction of the citronella grass while neem and yam bean were simple extracted by fermentation with ethyl alcohol. Toxicity test was evaluated in laboratory based on two methods of larvicidal bioassay: topical application method (contact poison) and feeding method (stomach poison). Larval mortality was observed daily and larval survivability was recorded until the survived larvae developed to pupae and adults. The study resulted that treatment of wood vinegar mixed with citronella grass showed the highest larval mortality by topical application method (50.0%) and by feeding method (80.0%). However, treatment of mixed wood vinegar and neem seed showed the longest pupal duration to 25 day and 32 days for topical application method and feeding method respectively. Additional, larval duration on treated M. domestica larvae was extended to 13 days for topical application method and 11 days for feeding method. Thus, the feeding method gave higher efficiency compared with the topical application method.

Keywords: housefly (Musca domestica L.), neem seed (Azadirachta indica), citronella grass (Cymbopogon nardus), yam bean seed (Pachyrhizus erosus), mortality

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Authors: Hajji Lobna, Chattaoui Mayssa, Regaieg Hajer, M'Hamdi-Boughalleb Naima, Rhouma Ali, Horrigue-Raouani Najet

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In this study, three local isolates of Trichoderma (Tr1: T. viride, Tr2: T. harzianum and Tr3: T. asperellum) were isolated and evaluated for their biocontrol effectiveness under in vitro conditions and in greenhouse. In vitro bioassay revealed a biopotential control against Fusarium oxysporum f. sp. radicis lycopersici and Meloidogyne javanica (RKN) separately. All species of Trichoderma exhibited biocontrol performance and (Tr1) Trichoderma viride was the most efficient. In fact, growth rate inhibition of Fusarium oxysporum f. sp. radicis lycopersici (FORL) was reached 75.5% with Tr1. Parasitism rate of root-knot nematode was 60% for juveniles and 75% for eggs with the same one. Pots experiment results showed that Tr1 and Tr2, compared to chemical treatment, enhanced the plant growth and exhibited better antagonism against root-knot nematode and root-rot fungi separated or combined. All Trichoderma isolates revealed a bioprotection potential against Fusarium oxysporum f. sp. radicis lycopersici. When pathogen fungi inoculated alone, Fusarium wilt index and browning vascular rate were reduced significantly with Tr1 (0.91, 2.38%) and Tr2 (1.5, 5.5%), respectively. In the case of combined infection with Fusarium and nematode, the same isolate of Trichoderma Tr1 and Tr2 decreased Fusarium wilt index at 1.1 and 0.83 and reduced the browning vascular rate at 6.5% and 6%, respectively. Similarly, the isolate Tr1 and Tr2 caused maximum inhibition of nematode multiplication. Multiplication rate was declined at 4% with both isolates either tomato infected by nematode separately or concomitantly with Fusarium. The chemical treatment was moderate in activity against Meloidogyne javanica and Fusarium oxysporum f. sp. radicis lycopersici alone and combined.

Keywords: trichoderma spp., meloidogyne javanica, Fusarium oxysporum f.sp.radicis lycopersici, biocontrol

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Authors: Michael Eickermann, Juergen Junk

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The Western honey (Apis mellifera) is threatened by a number of parasites, especially the devastating Varroa mite (Varroa destructor) is responsible for a high level of mortality over winter, e.g., in Europe and USA. While the use of synthetic pesticides or organic acids has been preferred so far to control this parasite, breeding strategies for less susceptible honey bees are in early stages. Hygienic behavior can be an important tool for controlling Varroa destructor. Worker bees with a high level of this behavior are able to detect infested brood in the cells under the wax lid during pupation and remove them out of the hive. The underlying processes of this behavior are only partly investigated, but it is for sure that hygienic behavior is heritable and therefore, can be integrated into commercial breeding lines. In a first step, breeding lines with a high level of phenotypic hygienic behavior have been identified by using a bioassay for accurate assessment of this trait in a long-term national breeding program in Luxembourg since 2015. Based on the artificial infestation of nucleus colonies with 150 phoretic Varroa destructor mites, the level of phenotypic hygienic behavior was detected by counting the number of mites in all stages, twelve days after infestation. A nucleus with a high level of hygienic behavior was overwintered and used for breeding activities in the following years. Artificial insemination was used to combine different breeding lines. Buckfast lines, as well as Carnica lines, were used. While Carnica lines offered only a low increase of hygienic behavior up to maximum 62.5%, Buckfast lines performed much better with mean levels of more than 87.5%. Some mating ends up with a level of 100%. But even with a level of 82.5% Varroa mites are not able to reproduce in the colony anymore. In a final step, a nucleus with a high level of hygienic behavior were build up to full colonies and located at two places in Luxembourg to build up a drone congregation area. Local beekeepers can bring their nucleus to this location for mating the queens with drones offering a high level of hygienic behavior.

Keywords: agiculture, artificial insemination, honey bee, varroa destructor

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Authors: Kiran Nawaz, Waheed Anwar, Sehrish Iftikhar, Muhammad Nasir Subhani, Ahmad Ali Shahid

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Plant growth promoting rhizobacteria (PGPR) have been extensively studied and applied for the biocontrol of many soilborne diseases. These rhizobacteria are very efficient against root rot and many other foliar diseases associated with solanaceous plants. These bacteria may inhibit the growth of various pathogens through direct inhibition of target pathogens or indirectly by the initiation of systemic resistance (ISR) which is active all over the complete plant. In the present study, 20 different rhizobacterial isolates were recovered from the root zone of healthy chili plants. All soil samples were collected from various chili-growing areas in Punjab. All isolated rhizobacteria species were evaluated in vitro and in vivo against Phytophthora capsici. Different species of Bacillus and Pseudomonas were tested for the antifungal activity against P. capsici the causal organism of Root rot disease in different crops together with chili. Dual culture and distance culture bioassay were carried out to study the antifungal potential of volatile and diffusible metabolites secreted from rhizobacteria. After seven days of incubation at 22°C, growth inhibition rate was recorded. Growth inhibition rate depended greatly on the tested bacteria and screening methods used. For diffusible metabolites, inhibition rate was 35-62% and 20-45% for volatile metabolites. The screening assay for plant growth promoting and disease inhibition potential of chili associated PGPR indicated 42-100% reduction in disease severity and considerable enhancement in roots fresh weight by 55-87%, aerial parts fresh weight by 35-65% and plant height by 65-76% as compared to untreated control and pathogen-inoculated plants. Pseudomonas flourescene, B. thuringiensis, and B. subtilis were found to be the most efficient isolates in inhibiting P. capsici radial growth, increase plant growth and suppress disease severity.

Keywords: rhizobacteria, chili, phytophthora, root rot

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Authors: S. Ben-Shabat, T. Turgeman, O. Leubinski, N. Roth-Bejerano, V. Kagan-Zur, Y. Sitrit

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The ectomycorrhizal (ECM) desert truffle Terfezia boudieri produces edible fruit bodies and forms symbiosis with its host plant Helianthemum sessiliflorum (Cistaceae) in the Negev desert of Israel. The symbiosis is vital for both partners' survival under desert conditions. Under desert habitat conditions, ECMs must form symbiosis before entering the dry season. To secure a successful encounter, in the course of evolution, both partners have responded by evolving special signals exchange that facilitates recognition. Members of the Cistaceae family serve as host plants for many important truffles. Conceivably, during evolution a common molecule present in Cistaceae plants was recruited to facilitate successful encounter with ectomycorrhizas. Arbuscular vesicular fungi (AM) are promiscuous in host preferences, in contrast, ECM fungi show specificity to host plants. Accordingly, we hypothesize that H. sessiliflorum secretes a chemotrophic-signaling, which is common to plants hosting ECM fungi belonging to the Pezizales. However, thus far no signaling molecules have been identified in ECM fungi. We developed a bioassay for chemotrophic activity. Fractionation of root exudates revealed a substance with chemotrophic activity and molecular mass of 534. Following the above concept, screening the transcriptome of Terfezia, grown under chemoattraction, discovered genes showing high homology to G proteins-coupled receptors of plant pathogens involved in positive chemotaxis and chemotaxis suppression. This study aimed to identify the active molecule using analytical methods (LC-MS, NMR etc.). This should contribute to our understanding of how ECM fungi communicate with their hosts in the rhizosphere. In line with the ability of Terfezia to form also endomycorrhizal symbiosis like AM fungi, analysis of the mechanisms may likewise be applicable to AM fungi. Developing methods to manipulate fungal growth by the chemoattractant can open new ways to improve inoculation of plants.

Keywords: chemotrophic signal, Helianthemum sessiliflorum, Terfezia boudieri, ECM

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Authors: Seyyed Payman Ziapour, Ahmadali Enayati, Sadegh Kheiri, Farzaneh Sahraei-Rostami, Reza Ali Mohammadpour, Mahmoud Fazeli-Dinan, Mohsen Aarabi, Fatemeh Asgarian, Seyed Hassan Nikookar, Mohammad Sarafrazi

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Rhipicephalus bursa (R. bursa) is a two-host ixodid tick with wide distribution in north of Iran especially in domestic animals of Mazandaran Province. The prolonged or incorrect use of chemical insecticides has led to build up of resistance in hard ticks in many areas of the world. Lack of basic information on resistance status of R. bursa was the reason behind this study to determine the susceptibility status of the species to lambda-cyhalothrin insecticide in Mazandaran Province. From May 2013 to March 2014, R. bursa ticks were collected on sheep, goat and cattle in different districts of Mazandaran Province. The engorged female ticks were reared in a controlled insectary for producing 12-18 days old larvae for larval packet test (LPT) bioassay against discriminant doses of lambda-cyhalothrin 5% EC (MAC SILAT®). 80% of ten pooled tick populations were susceptible to lambda-cyhalothrin as resistance ratios (RR50s) varied from 1 to 2.94 when compared with the most susceptible population NH-16. Only GK-12 and BF-6 populations (from plain areas of Galugah and Fereydunkenar Counties, respectively) were classified as resistant level I at LC50 level. Population NK-2 (from woodland areas of Kojour district in Nowshahr County) showed the highest resistance ratio of RR99 = 4.32 and 30% of tick populations were resistant at LC99 level. Our research showed initiation of lambda-cyhalothrin resistance in Rhipicephalus bursa populations in Mazandaran Province, Northern Iran. This is considered a warning to policy makers for disease control in the study area. This research is a part of the PhD thesis of SP. Ziapour by grant No. 92-89 in Student Research Committee, Mazandaran University of Medical Sciences, Iran.

Keywords: Rhipicephalus bursa, hard tick, lambda-cyhalothrin resistance, Iran

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Authors: Soledad Carinelli, Iñigo Fernández, José Luis González-Mora, Pedro A. Salazar-Carballo

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Mastitis is the most relevant inflammatory disease in cattle, affecting the animal health and causing important economic losses on dairy farms. This disease takes place in the mammary gland or udder when some opportunistic microorganisms, such as Staphylococcus aureus, Streptococcus agalactiae, Corynebacterium bovis, etc., invade the teat canal. According to the severity of the inflammation, mastitis can be classified as sub-clinical, clinical and chronic. Standard methods for mastitis detection include counts of somatic cells, cell culture, electrical conductivity of the milk, and California test (evaluation of “gel-like” matrix consistency after cell lysed with detergents). However, these assays present some limitations for accurate detection of subclinical mastitis. Currently, haptoglobin, an acute phase protein, has been proposed as novel and effective biomarker for mastitis detection. In this work, an electrochemical biosensor based on polydopamine-modified magnetic nanoparticles (MNPs@pDA) for haptoglobin detection is reported. Thus, MNPs@pDA has been synthesized by our group and functionalized with hemoglobin due to its high affinity to haptoglobin protein. The protein was labeled with specific antibodies modified with alkaline phosphatase enzyme for its electrochemical detection using an electroactive substrate (1-naphthyl phosphate) by differential pulse voltammetry. After the optimization of assay parameters, the haptoglobin determination was evaluated in milk. The strategy presented in this work shows a wide range of detection, achieving a limit of detection of 43 ng/mL. The accuracy of the strategy was determined by recovery assays, being of 84 and 94.5% for two Hp levels around the cut off value. Milk real samples were tested and the prediction capacity of the electrochemical biosensor was compared with a Haptoglobin commercial ELISA kit. The performance of the assay has demonstrated this strategy is an excellent and real alternative as screen method for sub-clinical bovine mastitis detection.

Keywords: bovine mastitis, haptoglobin, electrochemistry, magnetic nanoparticles, polydopamine

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Authors: Katie Emerson, Sara Perez-Ojalvo, Jim Komorowski, Danielle Greenberg

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The purpose of this study was to evaluate arginase activity levels in response to combinations of an inositol-stabilized arginine silicate (ASI; Nitrosigine®), L-arginine, and Inositol. Arginine acts as a vasodilator that promotes increased blood flow resulting in enhanced delivery of oxygen and nutrients to the brain and other tissues. ASI alone has been shown to improve performance on cognitive tasks. Arginase, found in human serum, catalyzes the conversion of arginine to ornithine and urea, completing the last step in the urea cycle. Decreasing arginase levels maintains arginine and results in increased nitric oxide production. This study aimed to determine the most effective combination of ASI, L-arginine and inositol for minimizing arginase levels and therefore maximize ASI’s effect on cognition. Serum was taken from untreated healthy donors by separation from clotted factors. Arginase activity of serum in the presence or absence of test products was determined (QuantiChrom™, DARG-100, Bioassay Systems, Hayward CA). The remaining ultra-filtrated serum units were harvested and used as the source for the arginase enzyme. ASI alone or combined with varied levels of Inositol were tested as follows: ASI + inositol at 0.25 g, 0.5 g, 0.75 g, or 1.00 g. L-arginine was also tested as a positive control. All tests elicited changes in arginase activity demonstrating the efficacy of the method used. Adding L-arginine to serum from untreated subjects, with or without inositol only had a mild effect. Adding inositol at all levels reduced arginase activity. Adding 0.5 g to the standardized amount of ASI led to the lowest amount of arginase activity as compared to the 0.25g 0.75g or 1.00g doses of inositol or to L-arginine alone. The outcome of this study demonstrates an interaction of the pairing of inositol with ASI on the activity of the enzyme arginase. We found that neither the maximum nor minimum amount of inositol tested in this study led to maximal arginase inhibition. Since the inhibition of arginase activity is desirable for product formulations looking to maintain arginine levels, the most effective amount of inositol was deemed preferred. Subsequent studies suggest this moderate level of inositol in combination with ASI leads to cognitive improvements including reaction time, executive function, and concentration.

Keywords: arginine, inositol, arginase, cognitive benefits

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Authors: Selvamuthukumaran Thirunavukkarasu, Arivudainambi Sundararajan

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The plant active molecules with their known mode of action are important leads to the development of newer insecticides. Lactone glycoside was identified earlier as the active principle in Cleistanthus collinus (Roxb.) Benth. (Fam: Euphorbiaceae). It possessed feeding deterrent, insecticidal and insect growth regulatory actions at varying concentrations. Deducing its mode of action opens a possibility of its further development. A no-choice leaf disc bioassay was carried out with lactone glycoside at different doses for different instars and Deterrence Indices were worked out. Using regression analysis concentrations imparting 10, 30 and 50 per cent deterrence (DI10, DI30 & DI50) were worked out. At these doses, effect on nutritional indices like Relative Consumption and Growth Rates (RCR & RGR), Efficiencies of Conversion of Ingested and Digested food (ECI & ECD) and Approximate Digestibility (AD) were worked out. The Relative Consumption and Growth Rate of control and lactone glycoside larva were compared by regression analysis. Regression analysis of deterrence indices revealed that the concentrations needed for imparting 50 per cent deterrence was 60.66, 68.47 and 71.10 ppm for third, fourth and fifth instars respectively. Relative consumption rate (RCR) and relative growth rate (RGR) were reduced. This confirmed the antifeedant action of the fraction. Approximate digestibility (AD) was found greater in treatments indicating reduced faeces because of poor digestibility and retention of food in the gut. Efficiency of conversion of both ingested and digested (ECI and ECD) food was also found to be greatly reduced. This indicated presence of toxic action. This was proved by comparing growth efficiencies of control and lactone glycoside treated larvae. Lactone glycoside was found to possess both feeding deterrent and toxic modes of action. Studies on molecular targets based on this preliminary site of action lead to new insecticide development.

Keywords: Spodoptera litura Fabricius, Cleistanthus collinus (Roxb.) Benth, feeding deterrence, mode of action

Procedia PDF Downloads 127

Authors: Karol Rodriguez-Peña, Martha L. Macias-Rubalcava, Leticia Rocha-Zavaleta, Sergio Sanchez

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A bioassay-guided study for anti-cancer compounds from endophytes of the Mexican medicinal plant Amphipteryygium adstringens resulted in the isolation of a streptomycete capable of producing a group of compounds with high cytotoxic activity. Microorganisms from surface sterilized samples of various sections of the plant were isolated and all the actinomycetes found were evaluated for their potential to produce compounds with cytotoxic activity against cancer cell lines MCF7 (breast cancer) and HeLa (cervical cancer) as well as the non-tumoural cell line HaCaT (keratinocyte). The most active microorganism was picked for further evaluation. The identification of the microorganism was carried out by 16S rDNA gene sequencing, finding the closest proximity to Streptomyces scabrisporus, but with the additional characteristic that the strain isolated in this study was capable of producing colorful compounds never described for this species. Crude extracts of dichloromethane and ethyl acetate showed IC50 values of 0.29 and 0.96 μg/mL for MCF7, 0.51 and 1.98 μg/mL for HeLa and 0.96 and 2.7 μg/mL for HaCaT. Scaling the fermentation to 10 L in a bioreactor generated 1 g of total crude extract, which was fractionated by silica gel open column to yield 14 fractions. Nine of the fractions showed cytotoxic activity. Fraction 4 was chosen for subsequent purification because of its high activity against cancerous cell lines, lower activity against keratinocytes. HPLC-UV-MS/ESI was used for the evaluation of this fraction, finding at least 10 different compounds with high values of m/z (≈588). Purification of the compounds was carried out by preparative thin-layer chromatography. The prevalent compound was Steffimycin B, a molecule known for its antibiotic and cytotoxic activities and also for its low solubility in aqueous solutions. Along with steffimycin B, another five compounds belonging to the steffimycin family were isolated and at this moment their structures are being elucidated, some of which display better solubility in water: an attractive property for the pharmaceutical industry. As a conclusion to this study, the isolation of endophytes resulted in the discovery of a strain capable of producing compounds with high cytotoxic activity that need to be studied for their possible utilization.

Keywords: amphipterygium adstringens, cytotoxicity, streptomyces scabrisporus, steffimycin

Procedia PDF Downloads 332

Authors: Mehdi Piri Ouchtapeh, Fariba Mehrkhou, Maryam Fourouzan

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The cabbage aphid, Bravicoryne brassicae (Hemiptera: Aphididae), is known as an economically important and oligophagous pest of different cole crops. The polyvolitine characteristics of B. brassicae resulted in resistance to insecticides. For this purpose, in this study, the sub-lethal concentration (LC25) of two insecticides, clothianidin and summer oil, on the life table parameters and population projection of cabbage aphid were studied at controlled condition (20±1 ℃, R.H. 60 ±5 % and a photoperiod of 16:8 h (L:D). The dipping method was used in bioassay and life table studies. Briefly, the leaves of cabbage containing 15 the same-aged (24h) adults of cabbage aphid (four replicates) were dipped into the related concentrations of insecticides for 10 s. The sub-lethal (LC25) obtained concentration were used 5.822 and 108.741 p.p.m for clothianidin and summer oil, respectively. The biological and life table studies were done using at least 100, 93 and 82 the same age of eggs for control, summer oil and clothianidin treatments respectively. The life history data of the greenhouse whitefly cohorts exposed to sublethal concentration of the aforementioned insecticides were analyzed using the computer program TWOSEX–MSChart based on the age-stage, two-sex life table theory. The results of this study showed that the used insecticides affected the developmental time, survival rate, adult longevity, and fecundity of the F1 generation. The developmental time on control, clothianidin and summer oil treatments was obtained (5.91 ± 0.10 days), (7.64 ± 0.12 days) and (6.66 ± 0.10 days), respectively. The sublethal concentration of clothianidin resulted in decreasing of adult longevity (8.63 ± 0.30 days), fecundity (14.14 ± 87 nymphs), survival rate (71%) and the life expectancy (10.26 days) of B. brassicae, as well. Additionally, usage of LC25 insecticides led to decreasing of the net reproductive rate (R0) of the cabbage aphid compared to summer oil and control treatments. The intrinsic rate of increase (r) (day-1) was decreased in F1 adults of cabbage aphid compared with other treatments. Additionally, the population projection results were accordance with the population growth rate of cabbage aphid. Therefore, the findings of this research showed that, however, both of the insecticides were effective on cabbage aphid population, but clothianidin was more effective and could be consider in the management of aforementioned pest.

Keywords: the cabbage aphid, sublethal effects, survival rate, population projection, life expectancy

Procedia PDF Downloads 38

Authors: A. K. Faizah, G. Vadamalai, S. K. Balasundram, W. L. Lim

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Pineapple (Ananas comosus) is a common crop in tropical and subtropical areas of the world. Malaysia once ranked as one of the top 3 pineapple producers in the world in the 60's and early 70's, after Hawaii and Brazil. Moreover, government’s recognition of the pineapple crop as one of priority commodities to be developed for the domestics and international markets in the National Agriculture Policy. However, pineapple industry in Malaysia still faces numerous challenges, one of which is the management of disease and pest. Red tip disease on pineapple was first recognized about 20 years ago in a commercial pineapple stand located in Simpang Renggam, Johor, Peninsular Malaysia. Since its discovery, there has been no confirmation on its causal agent of this disease. The epidemiology of red tip disease is still not fully understood. Nevertheless, the disease symptoms and the spread within the field seem to point toward viral infection. Bioassay test on nucleic acid extracted from the red tip-affected pineapple was done on Nicotiana tabacum cv. Coker by rubbing the extracted sap. Localised lesions were observed 3 weeks after inoculation. Negative staining of the fresh inoculated Nicotiana tabacum cv. Coker showed the presence of membrane-bound spherical particles with an average diameter of 94.25nm under transmission electron microscope. The shape and size of the particles were similar to tospovirus. SDS-PAGE analysis of partial purified virions from inoculated N. tabacum produced a strong and a faint protein bands with molecular mass of approximately 29 kDa and 55 kDa. Partial purified virions of symptomatic pineapple leaves from field showed bands with molecular mass of approximately 29 kDa, 39 kDa and 55kDa. These bands may indicate the nucleocapsid protein identity of tospovirus. Furthermore, a handheld sensor, Greenseeker, was used to detect red tip symptoms on pineapple non-destructively based on spectral reflectance, measured as Normalized Difference Vegetation Index (NDVI). Red tip severity was estimated and correlated with NDVI. Linear regression models were calibrated and tested developed in order to estimate red tip disease severity based on NDVI. Results showed a strong positive relationship between red tip disease severity and NDVI (r= 0.84).

Keywords: pineapple, diagnosis, virus, NDVI

Procedia PDF Downloads 762

Authors: Bagnoud-Velásquez Mariluz, Damergi Eya, Grandjean Dominique, Frédéric Vogel, Ludwig Christian

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The sustainability of algae to biofuel processes is seriously affected by the energy intensive production of fertilizers. Large amounts of nitrogen and phosphorus are required for a large-scale production resulting in many cases in a negative impact of the limited mineral resources. In order to meet the algal bioenergy opportunity it appears crucial the promotion of processes applying a nutrient recovery and/or making use of renewable sources including waste. Hydrothermal (HT) conversion is a promising and suitable technology for microalgae to generate biofuels. Besides the fact that water is used as a “green” reactant and solvent and that no biomass drying is required, the technology offers a great potential for nutrient recycling. This study evaluated the possibility to treat the water HT effluent by the growth of microalgae while producing renewable algal biomass. As already demonstrated in previous works by the authors, the HT aqueous product besides having N, P and other important nutrients, presents a small fraction of organic compounds rarely studied. Therefore, extracted heteroaromatic compounds in the HT effluent were the target of the present research; they were profiled using GC-MS and LC-MS-MS. The results indicate the presence of cyclic amides, piperazinediones, amines and their derivatives. The most prominent nitrogenous organic compounds (NOC’s) in the extracts were carefully examined by their effect on microalgae, namely 2-pyrrolidinone and β-phenylethylamine (β-PEA). These two substances were prepared at three different concentrations (10, 50 and 150 ppm). This toxicity bioassay used three different microalgae strains: Phaeodactylum tricornutum, Chlorella sorokiniana and Scenedesmus vacuolatus. The confirmed IC50 was for all cases ca. 75ppm. Experimental conditions were set up for the growth of microalgae in the aqueous phase by adjusting the nitrogen concentration (the key nutrient for algae) to fit that one established for a known commercial medium. The values of specific NOC’s were lowered at concentrations of 8.5 mg/L 2-pyrrolidinone; 1mg/L δ-valerolactam and 0.5 mg/L β-PEA. The growth with the diluted HT solution was kept constant with no inhibition evidence. An additional ongoing test is addressing the possibility to apply an integrated water cleanup step making use of the existent hydrothermal catalytic facility.

Keywords: hydrothermal process, microalgae, nitrogenous organic compounds, nutrient recovery, renewable biomass

Procedia PDF Downloads 384

Authors: Waheed Anwar, Kiran Nawaz, Muhammad Saleem Haider, Ahmad Ali Shahid, Sehrish Iftikhar

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The whitefly, Bemisia tabaci (Gennadius), is a complex insect species, including many cryptic species or biotypes. Whitefly causes damage to many ornamental and horticultural crops through directly feeding on phloem sap, resulting in sooty mould and critically decreases the rate of photosynthesis of many host plants. Biological control has emerged as one of the most important methods for the management of soil-borne plant pathogens. Among the natural enemies of insects different entomopathogenic fungi are mostly used as biological control of the pest. The purpose of this research was to find indigenous insect-associated fungi and their virulence against Bemisia tabaci. A detailed survey of cotton fields in sample collection was conducted during July and August 2013 from the central mixed zone of Punjab, Pakistan. For the isolation of T. longibrachiatum, sabouraud dextrose peptone yeast extract agar (SDAY) media was used and morphological characterization of isolated T. longibrachiatum was studied using different dichotomous keys. Molecular Identification of the pathogen was confirmed by amplifying the internal transcribed spacer region. Blastn analysis showed 100% homology with already reported sequences on the database. For these bioassays, two conidial concentrations 4 × 108/mL & 4 × 104/mL of T. longibrachiatum was sprayed in clip cages for nymph and adult B. tabaci respectively under controlled environmental conditions. The pathogenicity of T. longibrachiatum was tested on nymph and adult whitefly to check mortality. Mortality of B. tabaci at nymphal and adult stages were observed after 24-hour intervals. Percentage mortality of nymphs treated with 4 x 104/mL conidia of T. longibrachiatum was 20, 24, 36 and 40% after 48, 72, 96, 72, 96, 120 and 144 hours respectively. However, no considerable difference was recorded in percentage mortality of whitefly after 120 and 144 hours. There were great variations after 24, 48, 72 and 96 hours in the rate of mortality. The efficacy of T. longibrachiatum as entomopathogenic fungi was evaluated in adult and nymphal stages of whitefly. Trichoderma longibrachiatum showed maximum activity on nymphal stages of whitefly as compared to adult stages. The percentage of conidial germination was also recorded on the outer surface of adult and nymphal stages of B. tabaci. The present findings indicated that T. longibrachiatum is an entomopathogenic fungus against B. tabaci and many species of Trichoderma were already reported as an antagonistc organism against a wide range of bacterial and fungal pathogens.

Keywords: efficacy, Trichoderma, virulence, bioassay

Procedia PDF Downloads 241

Authors: Josiane N. Muller, Allan K. R. Galardo, Tatiane A. Barbosa, Evan P. Ferro, Wellington M. Dos Santos, Ana Paula S. A. Correa, Edinaldo C. Rego, Jose B. P. Lima

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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.

Keywords: Aedes aegypti, bioassay, larvicida, vector control

Procedia PDF Downloads 103

Authors: I. Ahumada, L. Ascar, C. Pedraza, J. Montecino

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It has been found that the addition of biosolids to soil is beneficial to soil health, enriching soil with essential nutrient elements. Although this sludge has properties that allow for the improvement of the physical features and productivity of agricultural and forest soils and the recovery of degraded soils, they also contain trace elements, organic trace and pathogens that can cause damage to the environment. The application of these biosolids to land without the total reclamation and the treated wastewater can transfer these compounds into terrestrial and aquatic environments, giving rise to potential accumulation in plants. The general aim of this study was to evaluate the bioavailability of bisphenol A (BPA), and 17 α-ethynyl estradiol (EE2) in a soil-biosolid system using wheat (Triticum aestivum) plant assays and a predictive extraction method using a solution of hydroxypropyl-β-cyclodextrin (HPCD) to determine if it is a reliable surrogate for this bioassay. Two soils were obtained from the central region of Chile (Lo Prado and Chicauma). Biosolids were obtained from a regional wastewater treatment plant. The soils were amended with biosolids at 90 Mg ha-1. Soils treated with biosolids, spiked with 10 mgkg-1 of the EE2 and 15 mgkg-1 and 30 mgkg-1of BPA were also included. The BPA, and EE2 concentration were determined in biosolids, soils and plant samples through ultrasound assisted extraction, solid phase extraction (SPE) and gas chromatography coupled to mass spectrometry determination (GC/MS). The bioavailable fraction found of each one of soils cultivated with wheat plants was compared with results obtained through a cyclodextrin biosimulator method. The total concentration found in biosolid from a treatment plant was 0.150 ± 0.064 mgkg-1 and 12.8±2.9 mgkg-1 of EE2 and BPA respectively. BPA and EE2 bioavailability is affected by the organic matter content and the physical and chemical properties of the soil. The bioavailability response of both compounds in the two soils varied with the EE2 and BPA concentration. It was observed in the case of EE2, the bioavailability in wheat plant crops contained higher concentrations in the roots than in the shoots. The concentration of EE2 increased with increasing biosolids rate. On the other hand, for BPA, a higher concentration was found in the shoot than the roots of the plants. The predictive capability the HPCD extraction was assessed using a simple linear correlation test, for both compounds in wheat plants. The correlation coefficients for the EE2 obtained from the HPCD extraction with those obtained from the wheat plants were r= 0.99 and p-value ≤ 0.05. On the other hand, in the case of BPA a correlation was not found. Therefore, the methodology was validated with respect to wheat plants bioassays, only in the EE2 case. Acknowledgments: The authors thank FONDECYT 1150502.

Keywords: emerging compounds, bioavailability, biosolids, endocrine disruptors

Procedia PDF Downloads 116

Authors: Gabriela Kalcikova, Igor Bosevski, Ula Rozman, Andreja Zgajnar Gotvajn

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Antibiotics are extensively used in human medicine and also in animal husbandry to prevent or control infections. Recently, a lot of attention has been put on veterinary antibiotics, because their global consumption is increasing and it is expected to be 106.600 tons in 2030. Most of veterinary antibiotics are introduced into the environment via animal manure, which is used as fertilizer. One of such veterinary antibiotics is tiamulin. It is used the form of fumarate for treatment of pig and poultry. It is used against prophylaxis of dysentery, pneumonia and mycroplasmal infections, but its environmental impact is practically unknown. Tiamulin has been found very persistent in animal manure and thus it is expected that can be, during rainfalls, transported into the aquatic environment and affect various organisms. For assessment of its environmental impact, it is necessary to evaluate its biodegradability and toxicity to various organisms from different levels of a food chain. Therefore, the aim of our study was to evaluate ready biodegradability and toxicity of tiamulin fumarate to various organisms. Bioassay used included luminescent bacterium Vibrio fischeri heterotrophic and nitrifying microorganisms of activated sludge, water flea Daphnia magna and duckweed Lemna minor. For each species, EC₅₀ values were calculated. Biodegradability test was used for determination of ready biodegradability and it provides information about biodegradability of tiamulin under the most common environmental conditions. Results of our study showed that tiamulin differently affects selected organisms. The most sensitive organisms were water fleas with 48hEC₅₀ = 14.2 ± 4.8 mg/L and duckweed with 168hEC₅₀ = 22.6 ± 0.8 mg/L. Higher concentrations of tiamulin (from 10 mg/L) significantly affected photosynthetic pigments content in duckweed and concentrations above 80 mg/L cause visible chlorosis. It is in agreement with previous studies showing significant effect of tiamulin on green algae and cyanobacteria. Tiamuline has a low effect on microorganisms. The lower toxicity was observed for heterotrophic microorganisms (30minEC₅₀ = 1656 ± 296 mg/L), than Vibrio fisheri (30minEC₅₀ = 492 ± 21) and the most sensitive organisms were nitrifying microorganisms (30minEC₅₀ = 183 ± 127 mg/L). The reason is most probably the mode of action of tiamulin being effective to gram-positive bacteria while gram-negative (e.g., Vibrio fisheri) are more tolerant to tiamulin. Biodegradation of tiamulin was very slow with a long lag-phase being 20 days. The maximal degradation reached 40 ± 2 % in 43 days of the test and tiamulin as other antibiotics (e.g. ciprofloxacin) are not easily biodegradable. Tiamulin is widely used antibiotic in veterinary medicine and thus present in the environment. According to our results, tiamulin can have negative effect on water fleas and duckweeds, but the concentrations are several magnitudes higher than that found in any environmental compartment. Tiamulin is low toxic to tested microorganisms, but it is very low biodegradable and thus possibly persistent in the environment.

Keywords: antibiotics, biodegradability, tiamulin, toxicity

Procedia PDF Downloads 159