Search results for: Caddisfly larvae
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 229

Search results for: Caddisfly larvae

199 Differentiating Third Instar Larvae of Three Species of Flies (Family: Sarcophagidae) of Potential Forensic Importance in Jamaica, Using Morphological Characteristics

Authors: Rochelle Daley, Eric Garraway, Catherine Murphy

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Crime is a major problem in Jamaica as well as the high number of unsolved violent crimes. The introduction of forensic entomology in criminal investigations has the potential to decrease the number of unsolved violent crimes through the estimation of PMI (post-mortem interval) or time since death. Though it has great potential, forensic entomology requires data from insects specific to a geographical location to be credibly applied in legal investigations. It is a relatively new area of study in the Caribbean, with multiple pioneer research opportunities. Of critical importance in forensic entomology is the ability to identify the species of interest. Larvae are commonly collected at crime scenes and a means of rapid identification is crucial. Moreover, a low-cost method is critical in countries with limited budget available for crime fighting. Sarcophagids are one of the most important colonisers of a carcass however, they are difficult to distinguish using morphology due to their similarities, however, there is a lack of research on the larvae of this family. This research contributes to that, having identified the larvae of three species from the family Sarcophagidae: Peckia nicasia, Peckia chrysostoma and Blaesoxipha plinthopyga; important agents in flesh decomposition. Adults of Sarcophidae are also difficult to differentiate, often requiring study of the genitalia; the use of larvae in species identification is important in such cases. Adult Sarcophagids were attracted using bottle traps baited with pig liver. These adults larviposited and the larvae were collected and colonises (generation 2 and 3) reared at room temperature for morphological work (n=50). The posterior ends of the larvae from segments 9 or 10 were removed and mounted posterior end upwards to allow study using a light microscope at magnification X200 (posterior cavity and intersegmental spine bands) and X640 (anterior and posterior spiracle). The remaining sections of the larvae were cleared in 10 % KOH and the cephalopharyngeal skeleton dissected out and measured at different points. The cephalopharyngeal skeletons show observable differences in the shapes and sizes of the mouth hooks as well as the length of the ventral cornua. The most notable difference between species is in the general shape of the anal segments and the shape of the posterior spiracles. Intersegmental spine bands of these larvae become less pigmented and visible as the larvae change instars. Spine bands along with anterior spiracle are not recommended as features for species distinction. Larvae can potentially be used to distinguish Sarcophagids to the level of species, with observable differences in the anal segments and the cephalopharyngeal skeletons. However, this method of identification should be tested by comparing these morphological features with other Jamaican Sarcophagids to further support this conclusion.

Keywords: 3rd instar larval morphology, forensic entomology, Jamaica, Sarcophagidae

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198 Assessment of Toxic Impact of Metals on Different Instars of Silkworm, Bombyx Mori

Authors: Muhammad Dildar Gogi, Muhammad Arshad, Muhammad Ahsan Khan, M. Sufian, Ahmad Nawaz, Mubashir Iqbal, Muhammad Junaid Nisar, Waleed Afzal Naveed

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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.

Keywords: heavy-metals, larval-instars, lethal-concentration, mortality, silkworm

Procedia PDF Downloads 183
197 Wireworms under the Sword of Damocles: Attraction to Maize Root Volatiles

Authors: Diana La Forgia, Jean Baptiste Thibord, François Verheggen

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Volatiles Organic Compound (VOCs) are one of the many features of defense used by plants in their eternal fight against pests. Their main role is to attract the natural enemies of the herbivores. But on another hand, they can be used by the same herbivores to locate plants while foraging. In an attempt to fill a gap of knowledge in a complex web of interactions, we focused on wireworms (Coleoptera:Elateridae). Wireworms whose larvae feed on roots are one of the most spread pests of valuable crops such as maize and potatoes, causing important economical damage. Little is known about the root compounds that are playing a role in the attraction of the larvae. In order to know more about these compounds, we compared four different maize varieties (Zea mays mays) that are known to have different levels of attraction, from weak to strong, for wireworms in fields. We tested the attraction of larvae in laboratory conditions in dual-choice olfactometer assays where they were offered all possible combinations of the four maize varieties. Contemporary, we collected the VOCs of each variety during 24h using a push-and-pull system. The collected samples were then analyzed by gas chromatography coupled with a mass spectrometer (GC-MS) to identify their molecular profiles. The choice of the larvae was dependent on the offered combination and some varieties were preferred to others. Differences were also observed in terms of quantitative and qualitative emissions of volatile profiles between the maize varieties. Our aim is to develop traps based on VOCs from maize roots to open a new frontier in wireworms management.

Keywords: integrated pest management, maize roots, plant defense, volatile organic compounds, wireworms

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196 Effect of Juvenile Hormone on Respiratory Metabolism during Non-Diapausing Sesamia cretica Wandering Larvae (Lepidoptera: Noctuidae)

Authors: E. A. Abdel-Hakim

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The corn stemborer Sesamia cretica (Lederer), has been viewed in many parts of the world as a major pest of cultivated maize, graminaceous crops and sugarcane. Its life cycle is comprised of two different phases, one is the growth and developmental phase (non-diapause) and the other is diapause phase which takes place at the last larval instar. Several problems associated with the use of conventional insecticides, have strongly demonstrated the need for applying alternative safe compounds. Prominent among the prototypes of such prospective chemicals are the juvenoids; i.e. the insect (JH) mimics. In fact, the hormonal effect on metabolism has long been viewed as a secondary consequence of its direct action on specific energy-requiring biosynthetic mechanisms. Therefore, the present study was undertaken essentially in a rather systematic fashion as a contribution towards clarifying metabolic and energetic changes taking place during non-diapause wandering larvae as regulated by (JH) mimic. For this purpose, we applied two different doses of JH mimic (Ro 11-0111) in a single (standard) dose of 100µg or in a single dose of 20 µg/g bw in1µl acetone topically at the onset of nondiapause wandering larvae (WL). Energetic data were obtained by indirect calorimetry methods by conversion of respiratory gas exchange volumetric data, as measured manometrically using a Warburg constant respirometer, to caloric units (g-cal/g fw/h). The findings obtained can be given in brief; these treated larvae underwent supernumerary larval moults. However, this potential the wandering larvae proved to possess whereby restoration of larval programming for S. cretica to overcome stresses even at this critical developmental period. The results obtained, particularly with the high dose used, show that 98% wandering larvae were rescued to survive up to one month (vs. 5 days for normal controls), finally the formation of larval-adult intermediates. Also, the solvent controls had resulted in about 22% additional, but stationary moultings. The basal respiratory metabolism (O2 uptake and CO2 output) of the (WL), whether un-treated or larvae not had followed reciprocal U-shaped curves all along of their developmental duration. The lowest points stood nearly to the day of prepupal formation (571±187 µl O2/gfw/h and 553±181 µl CO2/gfw/h) during un-treated in contrast to the larvae treated with JH (210±48 µl O2/gfw/h and 335±81 µl CO2/gfw/h). Un-treated (normal) larvae proved to utilize carbohydrates as the principal source for energy supply; being fully oxidised without sparing any appreciable amount for endergonic conversion to fats. While, the juvenoid-treated larvae and compared with the acetone-treated control equivalents, there existed no distinguishable differences between them; both had been observed utilising carbohydrates as the sole source of energy demand and converting endergonically almost similar percentages to fats. The overall profile, treated and un-treated (WL) utilized carbohydrates as the principal source for energy demand during this stage.

Keywords: juvenile hormone, respiratory metabolism, Sesamia cretica, wandering phase

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195 Effects of Gamma Radiation on Tomato Leafminer, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae)

Authors: Akın Kuyulu, Hanife Genç

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In present study, it was aimed to evaluate the gamma radiation impacts on tomato leaf miner at different biological stages. The laboratory colony of tomato leaf miner was used to set up the experiments. Different biological stages of the insects (eggs, 4th instars and pupae) were irradiated using Cobalt-60 at doses of 0 (control), 100 Gray (Gy), 200 Gy, 300 Gy and 400 Gy in Cos-44HH-N source, at dose rate of 480 Gy/h. After irradiation, the eggs were incubated until hatching; the mature larvae were reared to complete their developments. Adult emergences from irradiated pupae were also evaluated. The results showed that there were no egg hatching at all tested irradiation doses. Although, the pupal percentages of irradiated mature larvae were 54%, 15% and 8% at doses of 100 Gy, 200 Gy and 300 Gy respectively, there were no adult emergences from irradiated mature larvae. On the other hand, the adult emergences were observed from irradiated pupae, decreased as radiation doses increased along with malformed adult appearance. Male and female individuals were out crossed with laboratory reared adults. Fecundity was correlated with radiation doses.

Keywords: irradiation, tomato, tomato leafminer, Tuta absoluta

Procedia PDF Downloads 211
194 Bioefficacy of Ocimum sanctum on Survival, Development and Reproduction of Dengue Vector Aedes aegypti L. (Diptera: Culicidae)

Authors: Mohd Shazad, K. K. Gupta

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Vector borne diseases are a serious global concern. Aedes aegypti, the primary vector for viruses that cause dengue fever, dengue haemorrhagic fever, chikungunya and yellow fever is widespread over large areas of the tropics and subtropics. In last decade, diseases transmitted by Aedes aegypti are of serious concern. In past decade, number of cases of dengue fever, dengue hemorrhagic fever, and chikungunya has increased multifold. Present research work focused on impact of ethanol extract of Ocimum sanctum on dengue vector Aedes aegypti. 0-24 hr. old fourth instar larvae of lab-bred population of Aedes aegypti were exposed to ethanol leaf extract of Ocimum with concentrations ranging from 50 ppm to 400 ppm. Survival and development and the treated larvae and reproductive behaviour of the adults emerged from the treated larvae was evaluated. Our results indicated larvicidal potential of the leaf ethanol extract. The influence of the extract was dose dependent. 77.2% mortality was observed in the larvae exposed to 400 ppm for 24 hr. Treatment at lower concentrations revealed delayed toxicity. The larvae survived after treatment showed severe developmental anomalies. Consequently, there was the significant increase in duration of fourth instar larva. The L4 treated with 400-ppm extract moulted after 4.6 days; this was in sharp contrast to control where the larval period of the fourth instar lasts three days. The treated fourth instar larvae in many cases transformed into larva-pupa intermediates with the combination of larva, pupa characters. The larva-pupa intermediates had reduced life span and failed to moult successfully. The adults emerged from the larvae treated with lower doses had reduced reproductive potential. The females exhibited longer preoviposition period, reduced oviposition rate, abnormal oviposition behaviour and decreased fertility. Our studies indicated the possibility of the presence of JH mimic or JH analogue in the leaf ethanol extract of Ocimum. The present research work explored the potentials of Ocimum sanctum, also known as the queen of herbs, in integrated vector management programme of Aedes aegypti, which is a serious threat to human health.

Keywords: Aedes aegypti, development, mortality, Ocimum sanctum reproduction

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193 Response Development of larvae Portunus pelagicus to Artificial Feeding Predigest

Authors: Siti Aslamyah, Yushinta Fujaya, Okto Rimaldi

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One of the problems faced in the crab hatchery operations is the reliance on the use of natural feed. This study aims to analyze the response of larval development and determine the initial stages crab larvae begin to fully able to accept artificial feeding predigest with the help of probiotic Bacillus sp. The experiment was conducted in June 2014 through July 2014 at the location of the scale backyard hatcheries, Bojo village Mallusettasi sub-district, district Barru. This study was conducted in two stages larval rearing. The first stage is designed in a completely randomized design with 5 treatments and each with 3 repetitions, ie, without the use of artificial feeding; predigest feed given from zoea 1 - megalopa; predigest feed given since zoea 2 - megalopa; predigest feed given from zoea 3 - megalopa; and feed predigest given since zoea 4 - megalopa. The second stage of the two treatments, i.e. comparing artificial feeding without and with predigest. The results showed that the artificial feeding predigest able to replace the use of natural feed started zoea 3 generated based on the survival rate. Artificial feeding predigest provide a higher survival rate (16%) compared to artificial diets without predigest only 10.8%. However, feed predigest not give a different effect on the rate of development of stadia. Cell activity in larvae that received artificial feed predigest higher with RNA-DNA ratio of 8.88 compared with no predigest only 5:36. This research is very valuable information for crab hatchery hatchery scale households have limitations in preparing natural food.

Keywords: artificial feeding, development of stadia, larvae Portunus pelagicus, predigest

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192 Species Diversity of Coleoptera (Insecta: Coleoptera) Damaging Saxaul (Chenopodiáceae: Haloxylon spp.) in the Deserts Area of South-East Kazakhstan

Authors: B. Mombayeva

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In the deserts area of south east of Kazakhstan, 16 species of Coleoptera from 6 families and 12 genus of insects damaging Saxaul have been revealed. The vast number of species belong to the Cerambycidae familyCapricorn Beetle (4 species) and Hemlock Borer of Melanophila genus and 3 species of weevils and flea-beetles, and 1 species of coctsinelids and carrion beetle. Some of them cause appreciable harm, and sometimes very heavy damageto saxaul. According to food specialization they are divided into polyphages and - oligophages. According to the confinement to saxaul parts, registered beetles insects mainly feed on generative parts (11 species) and leaves (5 species). 9 species from them feed on roots, leaves and generative organs. They are scarablike beetle’s larvae (Apatophysismongolica Semenov., Tursmenigenavarentzovi Melg., Phytoecia (Opsilla) coerulescens Scopoli., Apatophysismongolica Semenov.), Jewel beetles (Julodis (s. Str.) Variolaris (Pallas), Sphenoptera (s. Str.) cuprina Motschulsky, S. (s. str.) exarata (Fischer), SphenopterapotaniniJak.) and some weevil (Barisartemisiae Hbst.). The larvae eat the roots and the imago - generative organs. Their feeding noticeably has its effect on the condition of saxaul. Beetles also slightlygnaw vegetative organs of plants. Among the harmful species the desert Capricorn Beetle Julodisvariolaris (Pallas) deserved attention. Its larvae live in the soil and cause harm to the roots of Saxaul and other pasture plants. In addition, the larvae of Sphenopterapotanini, S.punctatissima colonize the roots, trunk and branches of Haloxylon. In the spring Saxaul flowers are much damaged by Ladybeetle Bulaealichatchovi.

Keywords: saxaul, coleoptera, insecta, haloxylon

Procedia PDF Downloads 231
191 The Role of EDTA and EDDS in Reducing Metal Toxicity for Aquaculture Shellfish Perna canaliculus

Authors: Daniel R. McDougall, Martin D. de Jonge, Gordon M. Miskelly, Duncan J. McGillivray, Andrew G. Jeffs

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The chelating agent ethylenediaminetetraacetic acid (EDTA) is commonly added as a cure-all to seawater in aquaculture hatcheries around the world to reduce heavy metal toxicity, significantly improve the survival of larval shellfish, and to therefore improve the overall production efficiency of the aquaculture industry. However, EDTA is not a biodegradable chemical and is considered to be a persistent organic pollutant, which will accumulate in the environment over time. This makes the use of EDTA unsustainable environmentally, and therefore alternatives should be considered. Ethylenediaminedisuccinic acid (EDDS) is a biodegradable alternative to EDTA with very similar metal chelation properties. This study investigates the effect of EDTA and EDDS at two different concentrations, on metal concentrations found within developing New Zealand green-lipped mussel (Perna canaliculus) larvae. P. canaliculus is New Zealand’s main shellfish aquaculture species, providing a major export for New Zealand’s economy, with excellent potential for increased production in the near future. It is well known that the early stages of bivalve development are the most vulnerable to metal toxicity and P. canaliculus is no exception. The commercially used concentration (12 µmol L⁻¹) of EDTA added to P. canaliculus larval rearing tanks often increases the yield of D-larvae by over 80%. This concentration of EDTA and EDDS will be tested in this study, along with a lower concentration (3 µmol L⁻¹). After 48 hours of larval development, the D-larvae will be analyzed for heavy metal content with Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and heavy metal distribution with synchrotron X-ray Fluorescence Microscopy (XFM). In this study, we found that EDDS also improves the yield of P. canaliculus larvae and could be a viable alternative to EDTA in aquaculture. Furthermore, results suggest a higher concentration of chelating agent is more effective for improving the yield of developing P. canaliculus larvae. Metals with significant differences in concentration with the addition of EDTA were Cr, Cu, Zn, Cd and Pb (P < 0.05). We observed for the first time to the author’s best knowledge, metal distribution within 100 µm P. canaliculus D-larvae using synchrotron XFM and found changes in the distribution of metals with the addition of EDTA. XFM also has the potential to provide information about the chemical state of the metals within mussel larvae. This research provides greater insight into the reasons for the effectiveness of adding the chelating agent to aquaculture culture water, and a more environmentally conscious alternative to the currently used EDTA, which could be extremely valuable for the aquaculture industry.

Keywords: EDDS, EDTA, heavy metals, P. canaliculus, toxicity, water treatment

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190 Influence of the Use of Fruits Byproducts on the Lipid Profile of Hermetia illucens, Tenebrio molitor and Zophoba morio Larvae

Authors: Rebeca P Ramos-Bueno, Maria Jose Gonzalez-Fernandez, Rosa M. Moreno-Zamora, Antonia Barros Heras, Yolanda Serrano Alonso, Carolina Sanchez Barranco

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Insects are a new source of fatty acids (FA), so they are considered a sustainable and environmentally friendly alternative for both animal feed and the human diet, and furthermore, their harvesting/rearing require a low-tech and low capital investment. For that reason, lipids obtained by insect breeding open interesting possibilities with alimentary and industrial purposes, i.e., the production of biodiesel. Particularly, certain insect species, especially during the larval stage, contain high proportions of fat which is highly dependent on their feed and stage of development. Among them, Hermetia illucens larvae can be bred on food wastes to produce fat- and protein-rich raw materials for food by-product management. So, insects can act as excellent bioconverters of organic waste to nutrient-rich materials. In this regard, the aim of the study was to evaluate the effects of fruit byproducts on the FA compositions of Tenebrio molitor, Zophoba morio, and H. illucens larvae. Firstly, oil was extracted with the green solvent ethyl acetate, and FA methyl ester was obtained and analyzed by GC to show the FA profile. In addition, the triacylglycerol (TAG) profile was obtained by HPLC. Dehydrated watermelon, tomato, and papaya by-products, as well as wheat-based control feed, were assayed. High FA content was reached by Z. morio larvae fed with all fruits; however, no differences were shown in lipid profile with any change. It is worth highlighting that both Z. morio and H. illucens could be selected as the best candidates for biodiesel production due to their high content of saturated FA. On the other hand, T. molitor larvae showed a higher content of monounsaturated FA than control larvae, whereas the n-6 polyunsaturated FA content decreased in larvae fed with fruits. This result indicates that the improvement of the FA profile of Tenebrio can depend on both the type of feeding and the intended use. The lipid profile of H. illucens larvae fed with papaya and tomato showed a slight increase in the content of α-linoleic acid (ALA, 18:3n3). This FA is the precursor of docosahexaenoic acid (DHA, 22:6n3), which plays an important role as a component of structural lipids in cell membranes as well as in the synthesis of eicosanoids, protecting and resolving. Also, it was evaluated the TAG profile of Z. morio larvae due to their highest oil content. The results showed a high oleic acid (OA, 18:1n9) content, which displays modulatory effects in a wide range of physiological functions, having anti-inflammatory and anti-atherogenic properties. In conclusion, this study clearly shows that Z. morio and H. illucens larvae constitute an alternative source of OA- and ALA-rich oils, respectively, which can be devoted for food use, as well as for using in the food and pharmaceutical industries, with agronomic implications. Finally, although the profile of Z. morio was not improved with fruit feeding, this kind of feeding could be used due to its low environmental impact.

Keywords: fatty acids, fruit byproducts, Hermetia illucens, Zophoba morio, Tenebrio molitor, insect rearing

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189 Bioaccumulation and Forensic Relevance of Gunshot Residue in Forensically Relevant Blowflies

Authors: Michaela Storen, Michelle Harvey, Xavier Conlan

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Gun violence internationally is increasing at an unprecedented level, becoming a favoured means for executing violence against another individual. Not only is this putting a strain on forensic scientists who attempt to determine the cause of death in circumstances where firearms have been involved in the death of an individual, but it also highlights the need for an alternative technique of identification of a gunshot wound when other established techniques have been exhausted. A corpse may be colonized by necrophagous insects following death, and this close association between the time of death and insect colonization makes entomological samples valuable evidence when remains become decomposed beyond toxicological utility. Entomotoxicology provides the potential for the identification of toxins in a decomposing corpse, with recent research uncovering the capabilities of entomotoxicology to detect gunshot residue (GSR) in a corpse. However, shortcomings of the limited literature available on this topic have not been addressed, with the bioaccumulation, detection limits, and sensitivity to gunshots not considered thus far, leaving questions as to the applicability of this new technique in the forensic context. Larvae were placed on meat contaminated with GSR at different concentrations and compared to a control meat sample to establish the uptake of GSR by the larvae, with bioaccumulation established by placing the larvae on fresh, uncontaminated meat for a period of time before analysis using ICP-MS. The findings of Pb, Ba, and Sb at each stage of the lifecycle and bioaccumulation in the larvae will be presented. In addition, throughout these previously mentioned experiments, larvae were washed once, twice and three times to evaluate the effectiveness of existing entomological practices in removing external toxins from specimens prior to entomotoxicologyical analysis. Analysis of these larval washes will be presented. By addressing these points, this research extends the utility of entomotoxicology in cause-of-death investigations and provides an additional source of evidence for forensic scientists in the circumstances involving a gunshot wound on a corpse, in addition to advising the effectiveness of current entomology collection protocols.

Keywords: bioaccumulation, chemistry, entomology, gunshot residue, toxicology

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188 Liposomal Antihelmintics in Parasitology

Authors: Nina Ivanova

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More than one third of the population and animals are infected with parasitic helminths. It is especially difficult to cure the larval forms of parasites. The larvae of Hymenolepis nana invade the villi of the intestinal mucosa. Toxocara larvae can live in the liver, heart, lungs, brain, eyes, and pancreas. Commercial antiparasitic drugs cannot guarantee a 100% cure after a single course of treatment, because parasite larvae invade the villi of the intestinal mucosa and the anthelmintics do not reach and kill cestode larvae. The aim of this work was to conduct a study of liposomal antihelminthics on the laboratory animals under the certification received from the Committee on Bioethics and Deontology. It has been checked: 1) anthelmintic activity of the liposomal form of fenasal in experimental hymenolepidosis of white mice (larval stage - Hymenolepis nana). 2) anthelmintic activity of the liposomal form of albendazole in experimental toxocariasis of white mice (in the lungs at the stage of larval migration). Since some helminths cause hemolysis of erythrocytes as we used a mixture of polar lipids developed by us with antihemolytic activity to obtain liposomes Fenasal and albendazole were included in the liposome membrane in the ratio of anthelmintic: lipids 1:10. . The average size of liposomes was 180 nm, and the concentration of lipids in liposomes was 1%. The researches were carried out on white male mice who were infected with Hymenolepis Nana invasional eggs in a peroral way with a doze of 100 eggs per animal. On the 5th day after infection, a liposomal fenasal and commercial fenasal were administered orally for comparison. The animals were observed for 15 days. Before the introduction of liposomes and on the 3rd, 5th, 15th day after the administration of the drug, studies were carried out on the presence of helminths in the organs of animals. The liposomal fenasal, when administered orally, had an anthelmintic effect on Hymenolepis Nana cysticercoids at a dose of 25 mg/kg. The percentage of efficiency was 90.06, 91.36 96.85% on days 3, 5, 15, respectively. For comparison, the commercial activity was at a dose of 200 mg/ml, which is 8 times higher than the dose of liposomal fenasal. To evaluate the anthelmintic effect of the liposomal form of albendazole in experimental toxocariasis of white mice (toxascaris in the lungs at the stage of larval migration), studies were also carried out on white mice. The animals were infected with invasive eggs of Toxocara canis, orally, at a dose of 100 eggs per animal. On the 5th day after infection, the liposomal albendazole was administered orally. The efficacy of the study dosage form was determined by counting Larva mirgans larvae in the lungs. The results obtained showed that the liposomal albendazole had the greatest anthelmintic effect on Toxocara larvae at a dose of 2.0 mg/kg, which was 3.75 times less than the therapeutic one. At the same time, the percentage of efficiency was 93.75% on the 3rd day, and 98.66% on the 5th day.

Keywords: hymenolepis, Toxocara, larvae, liposomes

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187 Controlling the Growth and Development of Mosquito (Aedes aegypti) Using Testosterone

Authors: Brian F. Estidola, Alfredo A. Alcantara, Catherine del Cruz, Genelita S. Garcia

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This study aimed to investigate the effects of testosterone in the development and growth of Aedes aegypti as a main vector of dengue virus. There were three concentrations of testosterone: (0µM), (10µM), and (15µM) arranged randomly in two blocks. Each concentration houses 10 mosquitoes and monitored their development. The results showed that there were no significant differences on the effects of testosterone in emergence of larvae, mortality of eggs and larvae. However, it was shown that adults emerged from 15µM had a lower sex ratio than 10µM leading to the conclusion that there could be an optimal concentration of testosterone close to 10µM that could led to a high possibility of sex reversal of adult mosquitoes from female to male.

Keywords: mosquito, sex reversal, testosterone, ecdysterone

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186 Lucilia Sericata Netrin-A: Secreted by Salivary Gland Larvae as a Potential to Neuroregeneration

Authors: Hamzeh Alipour, Masoumeh Bagheri, Tahereh Karamzadeh, Abbasali Raz, Kourosh Azizi

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Netrin-A, a protein identified for conducting commissural axons, has a similar role in angiogenesis. In addition, studies have shown that one of the netrin-A receptors is expressed in the growing cells of small capillaries. It will be interesting to study this new group of molecules because their role in wound healing will become clearer in the future due to angiogenesis. The greenbottle blowfly Luciliasericata (L. sericata) larvae are increasingly used in maggot therapy of chronic wounds. This aim of this was the identification of moleculareatures of Netrin-A in L. sericata larvae. Larvae were reared under standard maggotarium conditions. The nucleic acid sequence of L. sericataNetrin-A (LSN-A) was then identified using Rapid Amplification of cDNA Ends (RACE) and Rapid Amplification of Genomic Ends (RAGE). Parts of the Netrin-A gene, including the middle, 3′-, and 5′-ends were identified, TA cloned in pTG19 plasmid, and transferred into DH5ɑ Escherichia coli. Each part was sequenced and assembled using SeqMan software. This gene structure was further subjected to in silico analysis. The DNA of LSN-A was identified to be 2407 bp, while its mRNA sequence was recognized as 2115 bp by Oligo0.7 software. It translated the Netrin-A protein with 704 amino acid residues. Its molecular weight is estimated to be 78.6 kDa. The 3-D structure ofNetrin-A drawn by SWISS-MODEL revealed its similarity to the Netrin-1 of humans with 66.8% identity. The LSN-A protein conduces to repair the myelin membrane in neuronal cells. Ultimately, it can be an effective candidate in neural regeneration and wound healing. Furthermore, our next attempt is to deplore recombinant proteins for use in medical sciences.

Keywords: maggot therapy, netrin-A, RACE, RAGE, lucilia sericata

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185 Management of Insect Pests Using Baculovirus Based Biopesticides in India

Authors: Mudasir Gani, Rakesh Kumar Gupta, Kamlesh Bali, Abdul Rouf Wani

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The gypsy moth (Lymantria obfuscata) and tent caterpillar (Malacosoma indicum) are serious pests that attack a wide range of fruit and forest trees in Jammu & Kashmir range of North-Western Himalayas in India. Investigations were carried out to isolate and bioprospect naturally occurring nucleopolyhedroviruses (NPVs) as potent biopesticides against these pests. The biological and molecular characterization of NPV isolates from different ecosystems was conducted, and the polh, lef-8 and lef-9 genes were sequenced and subjected to phylogenetic analysis. The L. obfuscata NPV was more closely related to the L. dispar NPV, whereas M. indicum NPV was more closely related to the M. californicum NPV in the NCBI taxonomy database. Among different isolates, Bhaderwah isolates exhibited highest virus activity (LD₅₀ = 250 POBs/larvae) and speed of kill (ST₅₀ = 6.80 days) against L. obfuscata whereas Mahor isolates proved most virulent against M. indicum, with lowest LD₅₀ (257 POBs/larva) and ST₅₀ (6.80 days). The in vivo mass production for highest productivity and quality revealed that the optimum yield was obtained when 3rd instar larvae were inoculated with a viral dose of 1.44 × 105 POBs/larva and allowed to incubate for nine days for L. obfuscata. However, for M. indicum larvae, a viral dose of 2.88 × 10⁶ POBs/larva and incubation period of 10 days were found optimum. It was found that harvesting of moribund larvae yields good quality NPV. The field application of L. obfuscata NPV and M. indicum NPV against the respective host populations on apple and willow with the pre-standardized dosage of 1 × 10¹² POBs/acre reduced the larval population density up to 25-63%.

Keywords: baculoviruses, biopesticides, Lymantria obfuscata, Malacosoma indicum

Procedia PDF Downloads 83
184 Effect of Four Medicinal Plant Extracts on Chickpea Leaf Miner Liriomyza cicerina (Rondani)

Authors: Sabraoui Abdelhadi, El Bouhssini Mustapha, Lhaloui Saadia, El Fakhouri Karim, Bouchelta Aziz

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The surveys carried out in 2014, 2015 in the regions of Abda- Doukala, Chaouia- Ouardigha, Zemour- Zair and Fes- Sais have confirmed that the leaf miner was the main insect pest attacking chickpea (Cicer arietinum L.) in Morocco. The grain yield losses caused by this pest could be more than 20% for winter planting and more than 42% for spring-sown crop. To reduce the chickpea leaf miner infestations, four essential oils, as biopesticide alternatives, were tested for their insecticidal effect on L. ciccerina, adults and larvae under laboratory conditions. In addition, we assessed the efficacy of these essential oils with and without adjuvant against this pest in comparison to three insecticides under field conditions. Mentha pulegium, with a dose of 33 µl/l of air caused 100% mortality on adults and larvae, after three hours and six hours of exposure, respectively. Eucalyptus showed 100% mortality on adults and larvae, with doses of 33 and 83 µl/l, after six and three hours of exposure, respectively. In the field conditions M. pulegium and E. globulus with adjuvant showed promising results compared with Abamectin, Azadirachtin and Spinetoram respectively. Essential oils could be used as one of the IPM components for the control of chickpea leaf miner.

Keywords: Liriomyza cicerina, chickpea, essential oils, insecticidal activity, Morocco

Procedia PDF Downloads 343
183 Laboratory Evaluation of Bacillus subtilis Bioactivity on Musca domestica (Linn) (Diptera: Muscidae) Larvae from Poultry Farms in South Western Nigeria

Authors: Funmilola O. Omoya

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Muscid flies are known to be vectors of disease agents and species that annoy humans and domesticated animals. An example of these flies is Musca domestica (house fly) whose adult and immature stages occur in a variety of filthy organic substances including household garbage and animal manures. They contribute to microbial contamination of foods. It is therefore imperative to control these flies as a result of their role in Public health. The second and third instars of Musca domestica (Linn) were infected with varying cell loads of Bacillus subtilis in vitro for a period of 48 hours to evaluate its larvicidal activities. Mortality of the larvae increased with incubation period after treatment with the varying cell loads. Investigation revealed that the second instars larvae were more susceptible to treatment than the third instars treatments. Values obtained from the third instar group were significantly different (P0.05) from those obtained from the second instars group in all the treatments. Lethal concentration (LC50) at 24 hours for 2nd instars was 2.35 while LC50 at 48 hours was 4.31.This study revealed that Bacillus subtilis possess good larvicidal potential for use in the control of Musca domestica in poultry farms.

Keywords: Bacillus subtilis, Musca domestica, larvicidal activities, poultry farms

Procedia PDF Downloads 385
182 Efficacy of Some Plant Extract against Larvae and Pupae of American Bollworm (Helicoverpa armigera) including the Effect on Peritropme Membrane

Authors: Deepali Lal, Sudha Summerwar, Jyoutsna Pandey

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The resistance of pesticide by the pest is an important matter of concern.The pesticide of plant origin having nontoxic biodegradable and environmentally friendly qualities. The frequent spraying of toxic chemicals is developing resistance to the pesticide. Leaf powder of the plants like Argimone mexicana and Calotropis procera is prepared, Different doses of these plant extracts are given to the Fourth in star stages of Helicoverpa armigera through feeding methods, to find their efficacy the experimental findings will be put under analysis using various parameters. The effect on paritrophic membrane is also studied.

Keywords: distillation plant, acetone, alcohol, pipette, castor leaves, grams pods, larvae of helicoverpa armigera, plant extract, vails, jars, cotton

Procedia PDF Downloads 281
181 Correlation between the Larvae Density (Diptera: Culicidae) and Physicochemical Characteristics of Habitats in Mazandaran Province, Northern Iran

Authors: Seyed Hassan Nikookar, Mahmoud Fazeli-Dinan, Seyyed Payman Ziapour, Ahmad-Ali Enayati

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Background: Mosquitoes look for all kinds of aquatic habitats for laying eggs. Characteristics of water habitats are important factors in determining whether a mosquito can survive and successfully completed their developmental stages. Physicochemical factors can display an important role in vector control programs. This investigate determined whether physicochemical factors differ between habitats can be effective in the larvae density in Mazandaran province. Methods: Larvae were collected by the standard dipper up to 350 ml for 15-20 minutes from fixed habitats in 16 villages of 30 townships, the specimens identified by morphological key. Water samples were collected during larval collection and were evaluated for temperature (°C), acidity (pH), turbidity (NTU), electrical conductivity (μS/cm), alkalinity (mg/l), total hardness (mg/l), nitrate (mg/l), chloride (mg/l), phosphate (mg/l), sulfate (mg/l) in selected habitats using standard methods. Spearman Correlation coefficient was used for analyze data. Results: Totally 7566 mosquito larvae of three genera and 15 species were collected of fixed habitats. Cx. pipiens was the dominant species except in villages of Tileno, Zavat, Asad Abad, Shah Mansur Mahale which An. maculipennis, Cx. torrentium were as the predominant species. Turbidity in Karat Koti, Chloride in Al Tappeh, nitrate, phosphate and sulfate in Chalmardi, electrical conductivity, alkalinity, total hardness in Komishan villages were significantly higher than other villages (P < 0.05). There were a significant positive correlation between Cx. pipiens and Electrical conductivity, Alkalinity, Total hardness, Chloride, Cx. tritaeniorhynchus and Chloride, whereas a significant negative correlation observed between Sulfate and Cx. perexiguss. Conclusion: The correlations observed between physicochemical factor and larval density, possibly can confirm the effect of these parameters on the breeding activities of mosquitoes, and could probability facilitate larval control programs by the handwork of such factors.

Keywords: anopheles, culex, culiseta, physicochemical, habitats, larvae density, correlation

Procedia PDF Downloads 233
180 The Chemical Composition and Larvicidal Activity of Essential Oils Derived from Piper Longepetiolatum and Piper Brachypetiolatum (Piperaceae) Against Aedes Aegypti Larvae (Culicidae) Were Investigated

Authors: Suelen C. Lima, André C. de Oliveira, Rosemary A. Roque

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Dengue is fatal arboviruses transmitted by the A. aegypti mosquito to more than 100 countries, for which the WHO estimates that 2.5 million people will be infected by these disease. The widespread of these diseases is due, among other factors, to the resistance that A. aegypti has to several commercial insecticides. On the other hand, natural products based on plants of the genus Piper (Piperaceae) are characterized by their insecticidal activities against mosquitoes. Piper longepetiolatum and Piper brachypetiolatum are species with wide distribution in the State of Amazonas. However, there is no investigation of phytochemical or biological of these plants against mosquitoes such as A. aegypti. The main of this study was to identify the chemical composition of the essential oil (EOs) from P. longepetiolatum and P. brachypetiolatum and to evaluate the biological activity against A. aegypti. The EOs were extracted by hydrodistillation from leaves (200 g) of P. longepetiolatum and P. brachypetiolatum and analyzed by GC-MS and GC-FID. The main compounds β-caryophyllene (99.9% of purity) and E-nerolidol (99.4% of purity) were purchased from Sigma-Aldrich® Brazil. The larvicidal activity of EOs (20 to 100 ppm), β-caryophyllene and E-nerolidol (10 to 50 ppm) was performed according to WHO protocol against A. aegypti larvae. The GC-MS and GC-FID analysis of EOs from P. longepetiolatum and P. brachypetiolatum indicated the majority presence of β-caryophyllene (35.42%) and E-nerolidol (49.79%), respectively. The results showed that all natural products presented larvicidal activity against A. aegypti. In this aspect, the OE from P. brachypetiolatum (LC50 of 15.51 ppm and LC90 of 22.79 ppm) was more active than the OE from P. longepetiolatum (LC50 of 47.17 ppm and LC90 of 69.60 ppm) (p < 0.05). Regarding of main compounds, E-nerolidol (LC50 of 9.50 ppm and LC90 of 23.89 ppm) showed higher larvicidal activity than the β-caryophyllene compound (LC50 of 79.00 ppm and LC90 of 230.91 ppm) (p < 0.05). The larvae treated with these natural products showed tremors and lethargic movements, suggesting that these natural products have neurotoxic action. These observations support studies to investigate the mechanism of action. This is the first record of the chemical composition and larvicidal activity of the EO from P. longepetiolatum and P. brachypetiolatum rich in β-caryophyllene and E-nerolidol against A. aegypti larvae.

Keywords: piperaceae, aedes, sesquiterpenes, biological control

Procedia PDF Downloads 39
179 Natural Enemies of the Fall Armyworm (Spodoptera frugiperda, Smith) and Comparing Neem Aqueous Extracts against Its Larvae in Gurage Zone, Central Ethiopia

Authors: Abera Hailu Degaga, Emana Getu Degaga

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Spodoptera frugiperda is an invasive insect pest that infests and feeds various crops, particularly affecting maize yields. However, nature has its own way of maintaining balance, and in this case, natural enemies play a crucial role in regulating the population of S. frugiperda. Locally available and easily prepared botanical sources, bio-pesticides, are also important. The objectives of the study were to investigate the natural enemies of S. frugiperda in the Gurage zone and to compare Neem aqueous extracts against its larvae in central Ethiopia. S. frugiperda larvae and egg masses were collected randomly from smallholder maize farms infested with pests between June and August 2023. Our findings revealed the existence of diverse types of parasitoids, predators, and entomopathogenic fungi associated with S. frugiperda. Notably, we documented three species of parasitoids, namely Exorista xanthaspis and Tachina spp. (Diptera: Tachinidae) and Charops annulipes (Hymenoptera: Ichneumonidae). All three species of parasitoids were recorded from Ethiopia for the first time. The overall parasitism rate was 5.3%, with individual rates ranging from 1.3 to 4%. Additionally, we identified ten species of predator insects from four different orders, including Hemiptera, Dermaptera, Coleoptera, and Mantodea, in the maize farms infested with S. frugiperda. Aqueous extract of Neem seed and leaf powder and green leaf exhibited similar mortality rates of S. frugiperda larvae at 72 hours even though there was a significant difference at 24 and 48 hours of the test. For effective management of S. frugiperda further research is necessary to fully exploit the potential of these natural enemies and additionally to use botanical source pesticides like Azadirachta indica.

Keywords: bio-pesticide, natural enemy, parasitoids, predators, Tachinid flies

Procedia PDF Downloads 23
178 Effects of Small Impoundments on Leaf Litter Decomposition and Methane Derived Carbon in the Benthic Foodweb in Streams

Authors: John Gichimu Mbaka, Jan Helmrich Martin von Baumbach, Celia Somlai, Denis Köpfer, Andreas Maeck, Andreas Lorke, Ralf Schäfer

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Leaf litter decomposition is an important process providing energy to biotic communities. Additionally, methane gas (CH4) has been identified as an important alternative source of carbon and energy in some freshwater food webs.Flow regulation and dams can strongly alter freshwater ecosystems, but little is known about the effect of small impoundments on leaf litter decomposition and methane derived carbon in streams. In this study, we tested the effect of small water storage impoundments on leaf litter decomposition rates and methane derived carbon. Leaf litter decomposition rates were assessed by comparing treatment sites located close to nine impoundments (Rheinland Pfalz state, Germany) and reference sites located far away from the impoundments.CH4 concentrations were measured in eleven impoundments and correlated with the δ13C values of two subfamilies of chironomid larvae (i.e. Chironomini and Tanypodinae). Leaf litter break down rates were significantly lower in study sites located immediately above the impoundments, especially associated with a reduction in the abundance of shredders. Chironomini larvae had the lower mean δ13C values (‒29.2 to ‒25.5 ‰), than Tanypodinae larvae (‒26.9 to ‒25.3 ‰).No significant relationships were established between CH4 concentrations and δ13C values of chironomids (p> 0.05).Mean δ13C values of chironomid larvae (mean: ‒26.8‰, range: ‒ 29.2‰ to ‒ 25.3‰) were similar to those of sedimentary organic matter (SOM) (mean: ‒28.4‰, range: ‒ 29.3‰ to ‒ 27.1‰) and tree leaf litter (mean: ‒29.8 ‰, range: ‒ 30.5‰ to ‒ 29.1‰). In conclusion, this study demonstrates that small impoundments may have a negative effect on leaf litter decomposition in forest streams and that CH4 has limited influence on the benthic food web in stream impoundments.

Keywords: river functioning, chironomids, Alder tree, stable isotopes, methane oxidation, shredder

Procedia PDF Downloads 701
177 Antifeedant Activity of Methanol and Hexane Extracts of Datura Innoxia (Mill.) (Solanaceae) in the Management of Spodoptera Litura (F.) (Lepidoptera: Noctuidae) Larvae

Authors: Vagisha Rawal, Anupam V. Sharma, Tarun Kumar Vats, Ashok Kumar Singh

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The antifeedant activity of methanol and hexane extract of leaves and seeds of Datura innoxia (Mill.) (Solanaceae) was evaluated against the 5th instar Spodoptera litura (F.) (Lepidoptera: Noctuidae) larvae in choice and no-choice leaf disc bioassays under laboratory conditions. These larvae when given a choice between the ‘control’ and ‘treated’ leaf discs in choice bioassays, consumed significantly (p ˂ 0.05) greater area of the ‘control’ leaf discs compared to those treated with the crude extracts of leaves and seeds of D. innoxia. The Antifeedant Index (AFI) for 5% concentration of the hexane extract of Datura seeds (DSHE) was 43.3% and 38.5% for methanol extract of Datura seeds (DSME). On the other hand, these values were 34.1% for the hexane extract of Datura leaves (DLHE), and 31.0% for the methanol extract of Datura leaves (DLME), respectively. In no-choice bioassays also, there was a significant (p˂0.05) reduction in the larval consumption of ‘treated’ leaf discs compared to the ‘control’ leaf discs. Maximum AFI was recorded at 5% concentration of the extracts of both the leaves and seeds with 47.7% for DSHE against 40.0% (DSME) and 39.4% for DLHE compared with 38.4% (DLME). Moreover, DSHE was found to have the maximum antifeedant effect irrespective of its concentration in comparison to the other crude extracts of leaves or seeds of D. innoxia. It is evident from these results that the crude methanol and hexane extracts of leaves and seeds of D. innoxia exhibited potent antifeedant activity against the 5th instar S. litura larvae. Also, the use of the bioactive compound(s) present in these extracts can prove to be an effective, eco-friendly, viable and sustainable component that can be integrated in IPM programs for the management of this economically important polyphagous insect pest in the Indian subcontinent.

Keywords: antifeedant activity, antifeedant index, datura innoxia, spodoptera litura

Procedia PDF Downloads 488
176 Zebrafish Larvae Model: A High Throughput Screening Tool to Study Autism

Authors: Shubham Dwivedi, Raghavender Medishetti, Rita Rani, Aarti Sevilimedu, Pushkar Kulkarni, Yogeeswari Perumal

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Autism Spectrum Disorder (ASD) is a complex neurodevelopmental disorder of early onset, characterized by impaired sociability, cognitive function and stereotypies. There is a significant urge to develop and establish new animal models with ASD-like characteristics for better understanding of underlying mechanisms. The aim of the present study was to develop a cost and time effective zebrafish model with quantifiable parameters to facilitate mechanistic studies as well as high-throughput screening of new molecules for autism. Zebrafish embryos were treated with valproic acid and a battery of behavioral tests (anxiety, inattentive behavior, irritability and social impairment) was performed on larvae at 7th day post fertilization, followed by study of molecular markers of autism. This model shows a significant behavioural impairment in valproic acid treated larvae in comparison to control which was again supported by alteration in few marker genes and proteins of autism. The model also shows a rescue of behavioural despair with positive control drugs. The model shows robust parameters to study behavior, molecular mechanism and drug screening approach in a single frame. Thus we postulate that our 7 days zebrafish larval model for autism can help in high throughput screening of new molecules on autism.

Keywords: autism, zebrafish, valproic acid, neurodevelopment, behavioral assay

Procedia PDF Downloads 130
175 Antifeedant Activity of Ageratum conyzoides (L.) (Asteraceae) Extracts against Diamondback Moth Plutella xylostella (L.) (Lepidoptera: Plutellidae)

Authors: Tarun Kumar Vats, Sanjiv Mullick, Vagisha Rawal, Ashok Kumar Singh

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Antifeedant activity of aqueous, methanolic and hexane crude extracts of powdered leaves of Ageratum conyzoides (L.) was evaluated against the last instar larvae of Plutella xylostella (L.), an oligophagous pest of Crucifer crops. Cauliflower leaf discs treated with different concentrations of extracts were provided to last instar larvae in both no-choice and choice bioassays under the standard laboratory conditions. All three extracts showed antifeedant effects in both the test conditions. In no-choice condition, hexane extract was found to significantly reduce the leaf area consumption at all the tested concentrations (0.5%, 1%, 2%, 3%, 4% and 5%). Also, aqueous and methanol extracts significantly reduced the leaf area consumption at different concentrations (P<0.05). In choice tests, effect of aqueous extract was significantly higher at 3%, 4% and 5% concentrations as compared to control. However, significant activities of methanol and hexane extracts were recorded even at lowest concentrations of 1% (P < 0.05). Complete feeding inhibition of larvae was observed at 2% concentration of hexane extract. Antifeedant index values (AFI) obtained were found to increase in a dose dependent manner, i.e. higher the concentration, more the activity. The results clearly indicate the potential of A. conyzoides extracts for its use in the integrated management of P. xylostella, which will be ecofriendly and sustainable.

Keywords: ageratum conyzoides, plutella xylostella, crucifer, antifeedant index

Procedia PDF Downloads 329
174 Isolation, Characterization, and Antibacterial Evaluation of Antimicrobial Peptides and Derivatives from Fly Larvae Sarconesiopsis magellanica (Diptera: Calliphoridae)

Authors: A. Díaz-Roa, P. I. Silva Junior, F. J. Bello

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Sarconesiopsis magellanica (Diptera: Calliphoridae) is a medically important necrophagous fly which is used for establishing the post-mortem interval. Dipterous maggots release diverse proteins and peptides contained in larval excretion and secretion (ES) products playing a key role in digestion. The most important mechanism for combating infection using larval therapy depends on larval ES. These larvae are protected against infection by a diverse spectrum of antimicrobial peptides (AMPs), one already known like lucifensin. Special interest in these peptides has also been aroused regarding understanding their role in wound healing since they degrade necrotic tissue and kill different bacteria during larval therapy. The action of larvae on wounds occurs through 3 mechanisms of action: removal of necrotic tissue, stimulation of granulation tissue, and antibacterial action of larval ES. Some components of the ES include calcium, urea, allantoin ammonium bicarbonate and reducing the viability of Gram positive and Gram negative bacteria. The Lucilia sericata fly larvae have been the most used, however, we need to evaluate new species that could potentially be similar or more effective than fly above. This study was thus aimed at identifying and characterizing S. magellanica AMPs contained in ES products for the first time and compared them with the common fly used L. sericata. These products were obtained from third-instar larvae taken from a previously established colony. For the first analysis, ES fractions were separate by Sep-Pak C18 disposable columns (first step). The material obtained was fractionated by RP-HPLC by using Júpiter C18 semi-preparative column. The products were then lyophilized and their antimicrobial activity was characterized by incubation with different bacterial strains. The first chromatographic analysis of ES from L. sericata gives 6 fractions with antimicrobial activity against Gram-positive bacteria Micrococus luteus, and 3 fractions with activity against Gram-negative bacteria Pseudomonae aeruginosa while the one from S. magellanica gaves 1 fraction against M. luteus and 4 against P. aeruginosa. Maybe one of these fractions could correspond to the peptide already known from L. sericata. These results show the first work for supporting further experiments aimed at validating S. magellanica use in larval therapy. We still need to search if we find some new molecules, by making mass spectrometry and ‘de novo sequencing’. Further studies are necessary to identify and characterize them to better understand their functioning.

Keywords: antimicrobial peptides, larval therapy, Lucilia sericata, Sarconesiopsis magellanica

Procedia PDF Downloads 339
173 A Study of the Prevalence of Trichinellosis in Domestic and Wild Animals for the Region of Sofia, Bulgaria

Authors: Valeria Dilcheva, Svetlozara Petkova, Ivelin Vladov

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Nemathodes of the genus Trichinella are zoonotic parasites with a cosmopolitan distribution. More than 100 species of mammals, birds and reptiles are involved in the natural cycle of this nematode. At present, T. spiralis, T. pseudospiralis, and T. britovi have been found in Bulgaria. The existence of natural wildlife and domestic reservoirs of Trichinella spp. can be a serious threat to human health. Three trichinella isolates caused human trichinella infection outbreaks from three regions of Sofia City Province were used for the research: sample No. 1 - Ratus norvegicus, sample No. 2 – domestic pig (Sus scrofa domestica), sample No. 3 - domestic pig (Sus scrofa domestica). Trichinella larvae of the studied species were isolated via digestive method (pepsin, hydrochloric acid, water) at 37ºC by standard procedure and were determined by gender (male and female) based on their morphological characteristics. As a reference trichinella species were used: T. spiralis, T. pseudospiralis, T. nativa and T. britovi. Single male and female larvae of the three isolates were crossed with single male and female larvae of the reference trichinella species as well as reciprocally. As a result of cross-breeding, offspring of muscular larvae with T. spiralis and T. britovi were obtained, while in experiments with T. pseudospiralis and T. nativa, trichinella larvae were not found in the laboratory mice. The results obtained in the control groups indicate that the trichinella larvae used from the isolates and the four trichinella species are infective. Also, the infective ability of the F1 offspring from the successful cross-breeding between isolates and reference species was investigated. Through the data obtained in the experiment was found that isolates No. 1 and No. 2 belong to the species T. spiralis, and isolate No. 3 belongs to the species T. britovi. The results were confirmed by PCR and real-time PCR analysis. Thus the presence and circulation of the species T. spiralis and T. britovi in Bulgaria was confirmed. Probably the rodents (rats) are involved in the distribution of T. spiralis in urban environment. The species T. britovi found in a domestic pig speaks of some contact with wild animals for which T. britovi is characteristic. The probable reason is that a large number of farmers in Bulgaria practice the free-range breeding of domestic pigs. Part of the farmers also used as food for domestic pigs waste products from the game (foxes, jackals, bears, wolves) and probably thus the infection was obtained. The distribution range of trichinella species in Bulgaria is not strictly outlined. It is believed that T. spiralis is most common in domestic animals and T. britovi and T. pseudospiralis are characteristic of wildlife. To answer the question whether wild and synanthropic animals are infected with the same or different trichinella species, which species predominate in nature and what their distribution among different hosts is, further research is required.

Keywords: cross-breeding, Sofia, trichinellosis, Trichinella britovi, Trichinella spiralis

Procedia PDF Downloads 156
172 Optimization of Headspace Solid Phase Microextraction (SPME) Technique Coupled with GC MS for Identification of Volatile Organic Compounds Released by Trogoderma Variabile

Authors: Thamer Alshuwaili, Yonglin Ren, Bob Du, Manjree Agarwal

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The warehouse beetle, Trogoderma variabile Ballion (Coleoptera: Dermestidae), is a major pest of packaged and processed stored products. Warehouse beetle is the common name which was given by Okumura (1972). This pest has been reported to infest 119 different commodities, and it is distributed throughout the tropical and subtropical parts of the world. Also, it is difficult to control because of the insect's ability to stay without food for long times, and it can survive for years under dry conditions and low-moisture food, and it has also developed resistance to many insecticides. The young larvae of these insects can cause damage to seeds, but older larvae prefer to feed on whole grains. The percentage of damage caused by these insects range between 30-70% in the storage. T. variabile is the species most responsible for causing significant damage in grain stores worldwide. Trogoderma spp. is a huge problem for cereal grains, and there are many countries, such as the USA, Australia, China, Kenya, Uganda and Tanzania who have specific quarantine regulations against possible importation. Also, grain stocks can be almost completely destroyed because of the massive populations the insect may develop. However, the purpose of the current research was to optimize conditions to collect volatile organic compound from Trogoderma variabile at different life stages by using headspace solid phase microextraction (SPME) coupled with gas chromatography-mass spectrometry (GC-MS) and flame ionization detection (FID). Using SPME technique to extract volatile from insects is an efficient, straightforward and nondestructive method. Result of the study shows that 15 insects were optimal number for larvae and adults. Selection of the number of insects depend on the height of the peak area and the number of peaks. Sixteen hours were optimized as the best extraction time for larvae and 8 hours was the optimal number of adults.

Keywords: Trogoderma variabile, warehouse beetle , GC-MS, Solid phase microextraction

Procedia PDF Downloads 96
171 A Wasp Parasitoids of Genus Cotesia (Hymenoptera: Braconidae) Naturally Parasitizing Pectinophora gossypiella (Saunders) on Transgenic Cotton in Indian Punjab

Authors: Vijay Kumar, G. K. Grewal, Prasad S. Burange

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India is one of the largest cultivators of cotton in the world. Among the various constraints, insect pests are posing a major hurdle to the success of cotton cultivation. Various bollworms, including the pink bollworm, Pectinophora gossypiella (Saunders), cause serious losses in India, China, Pakistan, Egypt, Brazil, tropical America, and Africa, etc. Bt cotton cultivars having Cry genes were introduced in India in 2002 (Cry1Ac) and 2006 (Cry1Ac+ Cry2Ab) for control of American, spotted, and pink bollworms. Pink bollworm (PBW) larvae infest flowers, squares, and bolls. Larva burrows into flowers and bolls to feed on pollen and seeds, respectively. It has a shorter lifecycle and more generations per year, so it develops resistance more quickly than other bollworms. Further, it has cryptic feeding sites, i.e., flowers and bolls/seeds, so it is not exposed to harsh environmental fluctuations and insecticidal applications. The cry toxin concentration is low in its feeding sites, i.e., seeds and flowers of cotton. The use of insecticide and Bt cotton is the primary control measure that has been successful in limiting the damage of PBW. But with the passage of time, it has developed resistance against insecticides and Bt cotton. However, the use of insecticides increases chemical control costs while causing secondary pest problems and environmental pollution. Extensive research has indicated that monitoring and control measures such as biological, cultural, chemical, and host plant resistance methods can be integrated for effective PBW management. The potential of various biological control organisms needs to be explored. The impact of transgenic cotton on non-target organisms, particularly natural enemies, which play an important role in pest control, is still being debated. According to some authors, Bt crops have a negative impact on natural enemies, particularly parasitoids. An experiment was carried out in the Integrated Pest Management Laboratory of the Department of Entomology, Punjab Agricultural University, Ludhiana, Punjab, India, to study the natural parasitization of PBW on Bt cotton in 2022. A large population of larvae of PBW were kept individually in plastic containers and fed with cotton bolls until the emergence of a parasitoid cocoon. The first cocoon of the parasitoid was observed on October 25, 2022. Symptoms of parasitization were never seen on larvae. Larvae stopped feeding and became inactive before the emergence of parasitoids for pupation. Grub makes its way out of larvae by making a hole in the integument, and immediately after coming out, it spins the cocoon. The adult parasitoid emerged from the cocoon after eight days. The parasitoids that emerged from the cocoon were identified as Cotesia (Braconidae: Hymenoptera) based on the features of the adult. Out of 475 larvae of PBW, 87 were parasitized, with 18.31% of parasitization. Out of these, 6.73% were first instar, 10.52% were second instar, and 1.05% were third instar larvae of PBW. No parasitization was observed in fourth instar larvae. Parasitoids were observed during the fag end of cropping season and mostly on the earlier instars. It is concluded that the potential of Cotesia may be explored as a biological control agent against PBW, which is safer to human beings, environment and non-taraltoget organisms.

Keywords: biocontrol, Bt cotton, Cotesia, Pectinophora gossypiella

Procedia PDF Downloads 54
170 A Comparative Laboratory Evaluation of Efficacy of Two Fungi: Beauveria bassiana and Acremonium perscinum, on Dichomeris eridantis Meyrick (Lepidoptera: Gelechiidae) Larvae, an Important Pest of Dalbergia sissoo

Authors: Gunjan Srivastava, Shamila Kalia

Abstract:

Dalbergia sissoo Roxb., (Family- Leguminosae; Subfamily- Papilionoideae), is an economically and ecologically important tree species having medicinal value. Of the rich complex of insect fauna, ten have been recognized as potential pests of nurseries and plantations. Present study was conducted to explore an effective ecofriendly control of Dichomeris eridantis Meyrick, an important defoliator pest of D. sissoo. Health and environmental concerns demanded devising a bio-intensive pest management strategy and employing ecofriendly measures. In the present laboratory bioassay two entomopathogenic fungi Acremonium perscinum and Beauveria bassiana were tested and compared for evaluating the efficacy of their seven different concentrations (besides control) against the 3rd, 4th and 5th instar larvae of D. eridantis, on the basis of mean percent mortality data recorded and tabulated for seven days after treatment application. Analysis showed that both treatments vary significantly among themselves. Also, variations amongst instars and duration with respect to their mortality were highly significant (p < .001). All their interactions were found to vary significantly. B. bassiana at 0.25x107 spores / ml spore concentration caused maximum mean percent mortality (62.38%) followed by mean percent mortality at its 0.25x106 spores / ml concentration (56.67%). Mean percent mortality at maximum spore concentration (0.054x107 spores / ml) and next highest spore concentration (0.054 x106 spores / ml) due to A. perscinum treatment were far less effective (mean percent mortality of 45.40% and 31.29%, respectively). At 168 hours mean percent mortality of larval instars due to both fungal treatment applications reached its maximum (52.99%) whereas, at 24 hours mean percent mortality remained least (5.70%). In both cases, treatments were most effective against 3rd instar larvae and least effective against 5th instar larvae. A comparative acccount of efficacy of B. bassiana and A. perscinum on the 3rd, 4th and 5th instar larvae of D. eridantis on 5th, 6th and 7th post treatment observation days after their application, on the basis of their median lethal concentrations (LC50) proved B. bassiana to be more potential microbial pathogen of the two fungal microbes, for all the three instars (3rd, 4th and 5th) of D. eridantis, on all the three days (5th, 6th and 7th post observation days after application of both treatments). Percent mortality of D. eridantis increased in a dose dependent manner. Koch’s Postulates tested positive, thus confirming the pathogenicity of B. bassiana against the larval instars of D. eridantis. LC90 values of 0.280x1011 spores/ml, 0.301x108 spores/ml and 0.262x108 spores/ml concentrations of B. bassiana were standardized which can effectively cause mortality of all the larval instars of D. eridantis in the field after 5th, 6th and 7th day of their application, respectively. Therefore, these concentrations can be safely used in nurseries as well as plantations of D. sissoo for effective control of D. eridantis larvae.

Keywords: Acremonium perscinum, Beauveria bassiana, Dalbergia sissoo, Dichomeris eridantis

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