Search results for: larvae/nymph mortality

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 219

Authors: Antonio B.Tangayan Jr., Hershey P. Mondejar, Pet Roey Pascual, Zeam Voltaire E. Amper

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A study on in vitro larvicidal acitivity of different levels of Madre de Cacao (Gliricidia sepium) concentrated crude ethanolic extract (CCEE) against horn fly larvae (Haematobia irritans) was conducted. The air-dried leaves of Gliricidia sepium were infused in a 1:3 ratio (w/v) using ethanol as solvent and concentrated in a rotary evaporator (60°C). A total of 120 larvae of Haematobia irritans were exposed in various concentration: 200, 400, 800 and 1000 ppm. Based on the result after 5 hours of exposure, CCE G. sepium extract at 200 ppm showed less effect with 30% mortality compared to 400 ppm, 800 ppm and 1000 ppm with 70%, 83%, and 100% mortality, respectively. Findings also revealed that CCE of G. sepium extract at 1000 ppm, 800 ppm, and commercial larvicide were comparable in causing mortality of H. irritans larvae from the first hour up to the fifth hours of exposure. However, on the fifth hour, 400 ppm was also found to be effective. This suggests that the higher the concentration of CCE G. sepium extract and the longer the time of exposure, the higher is the percentage mortality of the larvae. Thus, CCE G. sepium extract can be used as an alternative for commercial larvicide.

Keywords: horn fly, in vitro, larvicidal, Madre de Cacao

Procedia PDF Downloads 284

Authors: Ajuna B. Henry

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In recent decades, climate change has demanded biological pesticides; more Bt strains are being discovered worldwide, some containing novel insecticidal genes while others have been modified through molecular approaches for increased yield, toxicity, and wider host target. In this study, B. thuringiensis strain AH-2 (Bt-2) was isolated from the soil and tested for insecticidal activity against Aphis gossypii (Hemiptera: Aphididae) and Lepidoptera insect pests: fall webworm (Hyphantria cunea) and diamondback moth (Plutella xylostella). A commercial strain B. thuringiensis subsp. kurstaki (Btk), and a chemical pesticide, imidacloprid (for Hemiptera) and chlorantraniliprole (for Lepidoptera), were used as positive control and the same media (without bacterial inoculum) as a negative control. For aphidicidal activity, Bt-2 caused a mortality rate of 70.2%, 78.1% or 88.4% in third instar nymphs of A. gossypii (3N) at 10%, 25% or 50% culture concentrations, respectively. Moreover, Bt-2 was effectively produced in cost-effective (PB) supplemented with either glucose (PBG) or sucrose (PBS) and maintained high aphicidal efficacy with 3N mortality rates of 85.9%, 82.9% or 82.2% in TSB, PBG or PBS media, respectively at 50% culture concentration. Bt-2 also suppressed adult fecundity by 98.3% compared to only 65.8% suppression by Btk at similar concentrations but was slightly lower than chemical treatment, which caused 100% suppression. Partial purification of 60 – 80% (NH4)2SO4 fraction of Bt-2 aphicidal proteins purified on anion exchange (DEAE-FF) column revealed a 105 kDa aphicidal protein with LC50 = 55.0 ng/µℓ. For Lepidoptera pests, chemical pesticide, Bt-2, and Btk cultures, mortality of 86.7%, 60%, and 60% in 3rd instar larvae of P. xylostella, and 96.7%, 80.0%, and 93.3% in 6th instar larvae of H. cunea, after 72h of exposure. When the entomopathogenic strains were cultured in a cost-effective PBG or PBS, the insecticidal activity in all strains was not significantly different compared to the use of a commercial medium (TSB). Bt-2 caused a mortality rate of 60.0%, 63.3%, and 50.0% against P. xylostella larvae and 76.7%, 83.3%, and 73.3% against H. cunea when grown in TSB, PBG, and PBS media, respectively. Bt-2 (grown in cost-effective PBG medium) caused a dose-dependent toxicity of 26.7%, 40.0%, and 63.3% against P. xylostella and 46.7%, 53.3%, and 76.7% against H. cunea at 10%, 25% and 50% culture concentration, respectively. The partially purified Bt-2 insecticidal proteins fractions F1, F2, F3, and F4 (extracted at different ratios of organic solvent) caused low toxicity (50.0%, 40.0%, 36.7%, and 30.0%) against P. xylostella and relatively high toxicity (56.7%, 76.7%, 66.7%, and 63.3%) against H. cunea at 100 µg/g of artificial diets. SDS-PAGE analysis revealed that a128kDa protein is associated with toxicity of Bt-2. Our result demonstrates a medium and strong larvicidal activity of Bt-2 against P. xylostella and H. cunea, respectively. Moreover, Bt-2 could be potentially produced using a cost-effective PBG medium which makes it an effective alternative biocontrol strategy to reduce chemical pesticide application.

Keywords: biocontrol, insect pests, larvae/nymph mortality, cost-effective media, aphis gossypii, plutella xylostella, hyphantria cunea, bacillus thuringiensi

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Authors: Fatima Huda Hallak

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The current study aimed to evaluate the activity of bacterial insecticide Vertinic against the second star larvae of Trogoderma granarium (Everts) by four treatments: A, B, C, D, at seven concentrations: 0.001, 0.01, 0.1,1,10,100,1000 PPM. The mortality rate of larvae was 100% at concentrations 10 and 100 in treatments A and B after 24 hours and after 48 hours in treatment D at 1 PPM. The efficiency of treatment A was greater as compared to treatment B at all concentrations and all exposure times. The efficiency of treatment D was greater as compared to treatment C; for example, at 0.001, 0.01, 0.1, 1 PPM, after 120 hours, the Mortality rate of larve was 6.76, 13.33, 43.33, 100% in treatment D, which it was 0.00, 0.00, 23.33, 96.67%, respectively in the treatment C.

Keywords: bacterial insecticide, trogoderma granarium (everts), fourth star larvae, vertimic

Procedia PDF Downloads 51

Authors: A. Jatau, Q. Majeed, H. M. Bandiya

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The efficacy of five local spices, namely; Hot pepper (Capsicum annum L.), Black pepper (Piper guinese Schum and Thonn), Sweet basil (Occimum canum Sim), African nut-meg (Monodora myristica Dunal), and Ginger (Zingiber officianale Ross) with conventional insecticide against the D. maculatus was studied under ambient laboratory conditions. The plants were pulverized into powders and applied at the rate of 1.0, 2.0 and 3.0g per 25g of disinfected dried fish. The same amount of fish (25g) was treated with 5ml of 1.0, 2.0 and 3.0 percent solution of conventional insecticide (dichlorvos) and air dried for 2hrs. Ten newly hatched 1st instar larvae (24hrs old) were introduced into each powdered smoked fish in separate beakers. Untreated control was also set up. Observation on the mortality and development were recorded daily until the larvae pupated. Each of the treated smoked fish showed significant (p<0.05) effect on the larval mortality and development when compared with the control. The Piper guinense was as efficacious as dichlorvos in killing all the larvae (100%) at all concentrations before pupation. Ocimum Canunm gave the second best results (50.00, 63.33 and 100%), while the other three spices resulted in less than 50% mortalities at all rate of application. The spice powders were also observed to have extended the larval developmental period. Thus, the spices tested can be recommended for the control of D. maculatus.

Keywords: development, dermestes maculatus, insecticide, local spices, mortality

Procedia PDF Downloads 260

Authors: Muhammad Mamoon-Ur-Rashid, Gul Rehman

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Entomopathogenic fungi are considered effective bio-control agents for the management of a range of insect pests including red palm weevil. The research studies were conducted under laboratory and field conditions against 5th and 6th instars larvae and adults of [Rhynchophorus ferrugineus (Olivier)] at the faculty of Agriculture, Gomal University Dera Ismail Khan (KPK) Pakistan. The 5th instar larvae were used under field conditions whereas, the 6th instar larvae and newly emerged adults were used under lab conditions. Conidial suspensions were used at five different concentrations of 1×10⁴, 1×10⁵, 1×10⁶, 1×10⁷ and 1×10⁸, conidia per ml. The data were recorded on the mortality, total larval duration, weight of larvae, pre-pupal and pupal durations, percent pupal formation, pupal weight, percent adult emergence, and adult longevity (♂ and ♀) of red palm weevil. The B. bassiana had varying degrees of pathogenicity against different developmental stages of red palm weevil. The maximum larval duration (113.40 days) was noted when 5th instar larvae were treated with the maximum concentration (1 × 10⁸) of B. bassiana, whereas; the minimum total larval duration of 87.20 days was recorded on the lowest concentration (1 × 10⁴) of B. bassiana. The maximum pre-pual and pupal durations were noted at the maximum concentration. The maximum life span of adult male and females were noted at the lowest concentration, whereas; the minimum values were noted at the maximum concentration. The earliest mortality of red palm weevil was observed 1-day after treatment at higher concentrations of 1 × 10⁷ and 1 × 10⁸, whereas; it was recorded 3 and 4 days after treatment at lower concentrations of 1 × 10⁵ and 1 × 10⁴. At 10 days after treatment, the entomopathogenic fungus caused > 80% cumulative mortality of 5th and 6th instar larvae and adult weevils at the maximum concentrations which were more than double than those recorded at the lowest concentration. Overall, the 5th instar larvae of red palm weevils were most susceptible to the fungus compared to the 6th instar larvae and adult weevils. Based on current findings, it is suggested that entomopathogenic fungi could be used for the safer management of red palm weevil.

Keywords: entomopathogenic nematodes, mortality, red palm weevil, sub-lethal effects

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Authors: Hadjeb Ayoub, Mehaoua Med Seghir, Ouakid M. Laid

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Currently, chemical control is the only means used to control populations of the date moth (Ectomyelois ceratoniae) which is the most important and dangerous pest to palm groves in Algeria, conventional insecticides act faster, but their main drawback is it can’t be destroyed or degraded. In this context we conducted our work to explore the insecticidal activity of Spinpsad which is a bio-pesticide on the larval stages of Ectomyelois ceratoniae. The study of the effect of Spinosad on the mortality of different larval stages revealed that the doses used were significantly and positively correlated with mortality adjusted for different durations of exposure of larvae bio- pesticide. Lowest corrected mortality was observed in a short time and lethal in older larvae treated with the lowest concentration. While the higher mortality was observed in a longer duration of exposure in younger instars treated with the highest concentration.

Keywords: Ectomyelois ceratoniae, date palm, Spinosad, biological control, toxicology

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Authors: Hyoun Joong Kim, Jin Woo Jun, Sib Sankar Giri, Cheng Chi, Saekil Yun, Sang Guen Kim, Sang Wha Kim, Jeong Woo Kang, Se Jin Han, Se Chang Park

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Vibrio corallilyticus is a well-known pathogen of coral. It is also infectious to a variety of shellfish species, including Pacific oyster (Crassostrea gigas) larvae. V. corallilyticus is remained to be a major constraint in marine bivalve aquaculture practice, especially in artificial seed production facility. Owing to the high mortality and contagious nature of the pathogen, large amount of antibiotics has been used for disease prevention and control. However, indiscriminate use of antibiotics may result in food and environmental pollution, and development of antibiotic resistant strains. Therefore, eco-friendly disease preventative measures are imperative for sustainable bivalve culture. The present investigation proposes the application of bacteriophage (phage) as an effective alternative method for controlling V. corallilyticus infection in marine bivalve hatcheries. Isolation of phages from sea water sample was carried out using drop or double layer agar methods. The host range, stability and morphology of the phage isolates were studied. In vivo phage efficacy to prevent V. corallilyticus infection in oyster larvae was also performed. The isolated phages, named pVco-5 and pVco-7 was classified as a podoviridae and pVco-14, was classified as a siphoviridae. Each phages were infective to four strains of seven V. corallilyticus strains tested. When oyster larvae were pre-treated with the phage before bacterial challenge, mortality of the treated oyster larvae was lower than that in the untreated control. This result suggests that each phages have the potential to be used as therapeutic agent for controlling V. corallilyticus infection in marine bivalve hatchery.

Keywords: bacteriophage, Vibrio coralliilyticus, Oyster larvae, mortality

Procedia PDF Downloads 224

Authors: Gunjan Srivastava, Shamila Kalia

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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

Procedia PDF Downloads 225

Authors: Shabistana Nisar, Parvez Qamar Rizvi, Sheeraz Malik

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The potential advantage of nanotechnology is comparably marginal due to its unclear benefits in agriculture and insufficiency in public opinion. The nanotech products might solve the pesticide problems of societal concern fairly at acceptable or low risk for consumers and environmental applications. The deleterious effect of chemicals used on crops can be compacted either by reducing the existing active ingredient to nanosize or by plummeting the metals into nanoform. Considering the above facts, an attempt was made to determine the efficacy of nanoelements viz., Silver, Copper Manganese and Neem seed kernel extract (NSKE) for effective management of gram pod borer, Helicoverpa armigera infesting chickpea, being the most damaging pest of large number of crops, gram pod borer was selected as test insect to ascertain the impact of nanoparticles under controlled conditions (25-27 ˚C, 60-80% RH). The respective nanoformulations (0.01, 0.005, 0.003, 0.0025, 0.002, 0.001) were topically applied on 4th instar larvae of pod borer. In general, nanochemicals (silver, copper, manganese, NSKE) produced relatively high mortality at low dilutions (0.01, 0.005, 0.003). The least mortality was however recorded at 0.001 concentration. Nanosilver proved most efficient producing significantly highest (f₄,₂₄=129.56, p < 0.05) mortality 63.13±1.77, 83.21±2.02 and 96.10±1.25 % at 0.01 concentration after 2nd, 4th and 6th day, respectively. The least mortality was however recorded with nanoNSKE. The mortality values obtained at respective days were 21.25±1.50%, 25.20±2.00%, and 56.20±2.25%. Nanocopper and nanomanganese showed slow rate of killing on 2nd day of exposure, but increased (79.20±3.25 and 65.33±1.25) at 0.01 dilution on 3rd day, followed by 83.00±3.50% and 70.20±2.20% mortality on 6thday. The sluggishness coupled with antifeedancy was noticed at early stage of exposure. The change in body colour to brown due to additional melanisation in copper, manganese, and silver treated larvae and demalinization in nanoNSKE exposed larvae was observed at later stage of treatment. Thus, all the nanochemicals applied, produced the significant lethal impact on Helicoverpa armigera and can be used as valuable tool for its effective management.

Keywords: chickpea, helicoverpa armigera, management, nanoparticles

Procedia PDF Downloads 358

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 287

Authors: Eka Candra Lina, Indah Widhianingrum, Mita Eka Putri, Nur Afni Evalia, Muhammad Makky

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The emulsifiable concentrate (EC) and wettable powder (WP) of Piper aduncum and Tephrosia vogelii mixed formulations were tested for their activities in the laboratory and their effectiveness in the field against cabbage pest Plutella xyostella. Cabbage leaves soaked in six different mixed formulation concentrations were tested to 2ⁿᵈ instar larvae of P. xylostella with six replications. The observation was conducted everyday until larvae reached 4ᵗʰ instar stage. Correlation between concentration and larvae mortality was analyzed using probit (POLO-PC). The survived larvae was observed by looking at the growth and development, as well as the antifeedant effects. Field efficacy test was based on LC₉₅ value from laboratory test result. The experiment used a randomized block design with 5 treatments and 3 replications to test the populations of P. xylostella larvae and insecticide effectivity. The results showed that the EC and WP mixed formulations showed insecticidal activity against P. xylostella larvae, with LC₉₅ value of 0.35% and 0.37%, respectively. The highest antifeedant effect on EC mixed formulation was 85.01% and WP mixed formulation was 86.23%. Both mixed formulations also slowed the development of larvae when compared with control. Field effication result showed that applications of EC mixed formulation were able to restrain the population of P. xylostella, with effectivity value of 71.06%. Insecticide effectivity value of EC mixed formulation was higher than WP mixed formulation and Bacillus thuringiensis formulation.

Keywords: botanical insecticide, efficacy, emulsifiable concentrate (EC), Plutella xylostella, wettable powder (WP)

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Authors: Mohd Shazad, Kamal Kumar Gupta

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Aedes aegypti (Diptera: Culicidae), commonly known as tiger mosquito is the vector of dengue fever, yellow fever, chikungunya and zika virus. In the absence of any effective vaccine against these diseases, control the mosquito population is the only promising mean to prevent the diseases. Currently used chemical insecticides cause environmental contamination, high mammalian toxicity and hazards to non-target organisms, insecticide resistance and vector resurgence. Present research work aimed to explore the potentials of phytochemicals present in the Ocimum sanctum in management of mosquito population. The leaves of Ocimum were extracted with ethanol by ‘cold extraction method’. 0-24h old fourth instar larvae of Aedes aegypti were treated with the extract of concentrations 50ppm, 100ppm, 200ppm and 400ppm for 24h. Survival, growth and development of the treated larvae were evaluated. The adults emerged from the treated larvae were used for the reproductive fitness studies. Our results indicate 77.2% mortality in the larvae exposed to 400 ppm. At lower doses, although there was no significant reduction in the survival after 24h however, it decreased during subsequent days of observations. In control experiments, no mortality was observed. It was also observed that the larvae survived after treatment showed severe growth and developmental abnormalities. There was significant increase in larval duration. In control, fourth instar moulted into pupa after 3 days while larvae treated with 400 ppm extract were moulted after 4.6 days. Larva-pupa intermediates and the pupa-adult intermediates were observed in many cases. The adults emerged from the treated larvae showed impaired mating and oviposition behaviour. The females exhibited longer preoviposition period, reduced oviposition rate and decreased egg output. GCMS analysis of the ethanol extract revealed presence of JH mimics and intermediates of JH biosynthetic pathway. Potentials of Ocimum sanctum in integrated vector management programme of Aedes aegypti were discussed.

Keywords: Aedes aegypti, Ocimum sanctum, oviposition, survival

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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

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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

Procedia PDF Downloads 341

Authors: A. S. El Naas, K. S. Ganghish, A. O. Booker, O. Elwaer

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10 out of 40 camels examined at Tripoli slaughter places were infected with the camel nasal bot fly Cephalopenia titillator. The larvae occurred mainly in the nasopharynx and occasionally the larvae were found embedded between turbinated bones. The nasal cavity was congested and filled with mucus in which some larvae were entangled. This work is undertaken to determine the Prevalence of Cephalopenia titillator of camel slaughtered in Tripoli-Libya.

Keywords: Cephalopenia titillator, camels, Libya, larvae, nasopharynx

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Authors: Luz Hurtado, Rodrigo Rojas, Jaime Romero, Christopher Concha

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The culture of the marine fish red cusk eel Genypterus chilensis is currently considered a priority for Chilean aquaculture which is a Chilean native species of high gastronomic demand and market value. The microbiota was analyzed in terms of diversity and structure using massive Illumina sequencing. The analysis of alpha diversity was performed in samples of G. chilensis larvae of 6, 18 and 32 dph (days post-hatching) and it was observed that there were significant differences (P = 0.05) between the days of culture for the Chao1 index, being the larvae of 18 dph the one with the highest index followed by the larvae of 6 dph, The lowest value for this index was presented in larvae of 32 dph. There were no significant differences in larvae between the days of culture for the Shannon (P=0.0857) and Simpson (P=0.0714) indices. In general, the larvae of G. chilensis have high rates of diversity. When analyzing the beta diversity, a differentiation between the bacterial communities is observed depending on the day of the culture of the larvae. Considering the PCoA elaborated from the unweighted UniFrac statistic, the explained variance was 46.2% (PC1 29.2% and PC2 17.0%) and in the case of the PCoA elaborated with the weighted UniFrac statistic; the explained variance was 65.5% (PC1 41.8% and PC2 23.7%) these differences were significant based on the Permanova statistical analysis (P= 0.002 and 0.037 respectively). When analyzing the taxonomic composition of the microbiota of the larvae in the different days of culture it was observed that at the phyla level the most abundant in the larvae of 6 dph were Proteobacteria (57%) Verrucomicrobia (24%) and Firmicutes (14%), for the larvae of 18 dph the predominant phyla were Proteobacteria (90%), Dependientiae (5%), Actinobacteria (2%) and Plactomyces (2%), for the larvae of 32 dph the phyla that presented the highest relative abundance were Proteobacteria (57%), Firmicutes (29%), Verrucomicrobia (5%) and Actinobacteria (5%), when comparing the larvae between the days it was observed that the phylum Proteobacteria was the most abundant in the samples of larvae of 6, 18 and 32 dph being the larvae of 18 dph those that present the highest relative abundance, the larvae of 6 dph were those that presented the highest relative abundance for the phylum Verrucomicrobia and in the larvae of 32 dph was observed greater abundance of the phylum Firmicutes compared to the other days of larval culture. At the level of genera, those with the highest relative abundance in larvae of 6 dph were Rubritalea (30%), Psychrobacter (28%), staphylococcus (17%) and Ralstonia (10%), for the larvae of 18 dph the genera with the highest abundance were Psychrobacter (47%), Litoreibacter (13%), Nautella (9%) and Cohesibacter (8%), for the larvae of 32 dph the most abundant genera were Alloiococcus (25%), Dialister (14%), Neptunomonas (13%) and Piscirickettsia (11%). When observing the taxonomic composition of the larvae between the days of larval culture, it is observed that there are differences between them.

Keywords: microbiota, diversity, G. Chilensis, larvae

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Authors: Zunnu Raen Akhtar

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Brevicoryne brassicae, aphid feeds on cabbage and Brassica sp. as preferred host. Brassica plants usually ripen when maize starts growing in multiple cropping systems. Experiment was conducted to observe suitability of B. brassicae by rearing it on maize as host. In a tritrophic eco-system, predator coccinellids can be found in the fields of brassica and maize. This experiment emphasized on issue of aphids growing incidence in a cropping system. Brassica is sown and harvested earlier than maize and is attacked by aphids, while maize is also attacked by aphids. Five mortality tests were conducted of B. brassicae fed on maize. Out of five mortality tests, 3 tests were conducted using 1st instar, while in two mortality tests, 2nd instars of aphids were used. Mortality tests revealed that first instar mortality was quite high on the second day, while in second instar larvae mortality was delayed up to third to the fourth day. These experiments reveal that aphids can use maize as substitute host at later instars as compared to young ones. These experiments can be foundation for studying further crop-insect interaction and sampling techniques used for this purpose.

Keywords: host suitability, B. brassicae, maize, tritrophic interaction

Procedia PDF Downloads 194

Authors: Futo Asano, Yusuke Yatsushiro, Hirokuni Miyamoto, Hiroaki Kodama

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A thermophile-fermented compost is produced using small fishes, crabs, and shrimps under a high temperature (approximately 75℃) by fermentation-associated self-heating. This compost has been used as a feed additive for pigs and hens in Japan, and the fecundity of this livestock is enhanced. Firmicutes is a dominant phylum in the microbial composition of the compost. We first reported that improvement of female larval fitness of Hercules beetle can be achieved by amendment of this compost to the humus. When the 90-d-old larvae were reared for subsequent 72 days in the humus with this compost, the growth of female larvae was significantly enhanced when compared with the growth of female larvae in the humus without the compost. In contrast, the growth of male larvae in the compost-free humus was the same as the larvae grow in the compost-amended humus. The bacterial composition of the feces of larvae was determined at 0 days and 46 days after transfer to the humus with or without the compost. The most dominant bacterium in the feces was Xylanimonas. Interestingly, the growth improvement of female larvae was associated with an increased abundance of Mollicutes in the fecal samples. These results indicate that the compost act as a probiotic material for enhancing the female larvae growth by supporting Mollicutes. Here, we tried to isolate Mollicutes from the contents of the midgut and hindgut of the 3rd instar female larvae of the Hercules beetle. These gut contents were spread onto a selective agar medium for Mollicutes (PPLO agar broth, BD Difco, NJ, USA). Although we isolated none of the Mollicutes until now, several bacteria that are closely related to Xylanimonas and Luteimicrobium were isolated. These isolates have xylanase and glucanase (CMCase) activities. We show the gut bacterial profiles of larvae and discuss how the fitness of female larvae of the Hercules beetle is improved by the compost.

Keywords: compost, beetle, mollicutes, woody biomass

Procedia PDF Downloads 84

Authors: Guler D. Uzel, Andrew G. Parker, Robert L. Mach, Adly Abd-Alla

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Several tsetse fly species (Diptera: Glossinidae) in natural or colonized populations can be infected with the salivary gland hypertrophy virus (SGHV), a circular dsDNA virus (Hytrosaviridae). The virus infection is mainly asymptomatic but, in some species under certain conditions, the infection can produce salivary gland hypertrophy (SGH) symptoms. In the laboratory colonized tsetse, flies with SGH have reduced fertility, which negatively affects colony performance. Therefore, a high prevalence of SGH in insect mass rearing represents a major challenge for tsetse control using the sterile insect technique. The main objective of this study is to analyze the impact of Glossina pallidipes SGHV infection in various tsetse species on mortality and productivity and its impact on the symbiotic bacteria. Hypertropied salivary glands (SG) were collected from G. pallidipes into phosphate buffered saline (PBS) to prepare suspension; 2 µl aliquots were injected into adults of several tsetse species (G. pallidipes (Gp), G. p. gambiensis (Gpg), G. brevipalpis (Gb), G. morsitans morsitans (Gmm), G. morsitans centralis (Gmc) and G. fuscipes (Gf)) and the change in virus and symbiont titers were analyzed using qPCR. The development of SGH in the F1 was detected by dissection 10 days after emergence and virus infection was confirmed by PCR. The impact of virus infection on fly mortality and productivity was recorded. 2 µl aliquots were also injected into 3rd instar larvae of the different species and the adult SGs assayed by PCR for virus. Virus positive SGs from each species were homogenized in PBS and pooled within species for injection into larvae of the same species. Flies injected with PBS were used as control. Injecting teneral flies with SGHV caused increasing virus titer over time in all species but no SGH was detected. Dissection of the F1 also showed no development of SGH except in Gp (the homologous host). Injection of SGHV did not have any impact on the prevalence of the tsetse symbionts, but an increase in Sodalis titer was observed correlated with fly age regardless of virus infection. The virus infection had a negative impact on productivity and mortality. SGHV injection into larvae of the different species produced SGHV infected glands in the adults determined by PCR with a rate of 60%, 27%, 16%, 7% and 7% for Gp, Gf, Gpg, Gmm and Gmc, respectively. Virus positive SGs observed in the heterologous species were smaller than SGH found in Gp. No virus positive SG was detected by PCR in Gb and no SGH was observed in any adults except in Gp. Injecting virus suspension from the virus positive SGs into conspecific larvae did not produce any adults with infected SGs (except in Gp). SGHV can infect all tested tsetse species. Although the virus can infect and increase in titer in other tsetse species and affect fly mortality and productivity, no vertical virus transmission was observed in other tsetse species with might indicate a transmission barrier in these species, and virus collected from flies injected as larvae was not infective by injection.

Keywords: DNA viruses, glossina, hytrosaviridae, symbiotic bacteria, tsetse

Procedia PDF Downloads 216

Authors: Tadesse Kebede, Orkun Baris Kovanci

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Tomatoes leaf minor (Tutaabasoluta) is one of the most economically important insect pest in tomatoes production. The use of biological control such as entomopathogen fungi isolates would be a long-term and cost-effective solution to control insects pest. Therefore, identifying the most virulent and pathogenic entomopathogen fungi is one of the basic requirements for effective management options to combat Tomatoes leaf minor (Tutaabasoluta). Furthermore, the pathogenicity and virulence difference among entomopathogenfungus strains is not widely well investıgated. The current study was therefore initiated to test the pathogenicity of some entomopathogenic fungus isolates against Tutaabsoluta. The experiment was conducted at Bursa Uludag University, Agiculutre faculty, horticulture department glasshouse in 2020/2021. Tutabasoluta adult were collected, and masslarvae were reared in a growth chamber. Then, ten third instar larvae were inoculated with four entomopathogen fungi isolates (Beuaveriabassania Ak-10, Beuaveriabassania Ak-14, Metarhziumanisoplai Ak-11, and Metarhziumanisoplai Ak-12) with different inoculum suspension (0, 1x10⁶, 1x10⁷,,4 × 10⁸, 4× 10⁹ and 1×10¹⁰ conidia /ml) in a factorial experiment arranged in randomized complete block design with three replication. Mortality data assessment was done on the 3rd, 5thand 7th days after treatment and analyzed. The analysis of variance for mortality rate revealed significant variations (p<0.05) among entomoptahogen fungi isolates and conidia concentrations. The results revealed thatMetarhziumanisoplai Ak-12was found to show the lowest mortality percentage80.77%, highest LC50 2.3x108, and the longest incubation period, LT50, 4.9 and LT90, 9.9daysand considered to be less pathogenic fungi. On the other hand, Beuaveriabassania Ak-10 isolate showed the highest mortality percentage, 91%, and the lowest LT50, 4, and LT90, 7.6 values at 1×10¹⁰ conidia /ml, followed by Beuaveriabassania Ak-14 and being considered as the most aggressive bio-agent. Metarhziumanisoplai Ak-11 was determined as moderately virulent, having a mortality rate 27-81%. Results also revealed that among conidia concentrations, 1x10⁹ and 1x10¹⁰ suspensions is the most effective, while 1x10⁶ conidia/ml concentration is the least effective. Hence, results indicated that EPF tested were effective against T. absoluta larvae. As the current work revealed the potential variation among entomopathogen fungi isolates and concentration against third instar larvae.

Keywords: tuta absoluta, tomato, metarhizium anisopliae, beauveria bassiana, biological control

Procedia PDF Downloads 128

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

Procedia PDF Downloads 244

Authors: N. Adjlane, N. Haddad

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The American foulbrood is one of the most serious diseases that may affect brood of larvae and pupae stages. The causative organism is a gram positive bacterium Paaenibacillus larvae. American foulbrood infected apiaries suffer from severe economic losses, resulting from significant decreases in honeybee populations and honey production. The aim of this study was to detect Paenibacillus larvae in the remains collected at the bottom of the hive from the suspected hives by direct PCR and culture growth. A total of 56 suspected beehive wax debris samples collected in 40 different apiaries located in the central region of Algeria. MYPGP the culture medium is used during all the identifications of the bacterium. After positive results on samples, biochemical confirmation tests (test of catalase, presence hydrolysis of casein) and microscopic (gram stain) are used in order to verify the accuracy of the initial results. The QIAamp DNA Mini Kit is used to identify the DNA of Paaenibacillus larvae. Paaenibacillus larvae were identified in 14 samples out of 16 by the PCR. A suspected culture-negative sample was found positive through evaluation with PCR. This research is for the bacterium Paaenibacillus larvae in the debris of the colony is an effective method for diagnosis of the pathology of American foulbrood.

Keywords: Paenibacillus larvae, honeybee, PCR, microbiological method

Procedia PDF Downloads 411

Authors: Besma Hamrouni Assadi, Mohamed Sadok Belkadhi

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The whitefly Bemisia tabaci is a cosmopolitan insect that causes serious damage to greenhouse crops. It is increasingly recognized that the use of biological control means such as entomopathogenic fungi presents a sustainable solution to integrated pest management programs. In order to reduce the use of chemical pesticides, Beauveria bassiana strain R444 was tested against eggs and second, third and fourth instar larvae of B. tabaci in a geothermal tomato greenhouse in southern Tunisia. This entomopathogenic fungus was compared to a chemical pesticide Imidacloprid and an untreated control. We found significant mortality of individuals caused by B. bassiana comparable to that caused by the chemical pesticide. After four weeks of follow-up, this fungus causes a mortality of eggs and larvae of B. tabaci that exceeds 60%. It shows that the use of entomopathogenic fungi can help reduce the use of pesticides to control B. tabaci on geothermal crops.

Keywords: entomopathogenic fungi, Bemisia tabaci, geothermal greenhouse, integrated pest management programs

Procedia PDF Downloads 104

Authors: RuiJin Zhang, HengJiang Cai, JinSong Gui

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It takes Patinopecten yessoensis larvae more than 20 days from spawning to settlement. Due to the natural environmental factors such as current, Patinopecten yessoensis larvae are transported to a distance more than hundreds of kilometers, leading to a high instability of their spatial and temporal distribution and great difficulties in the natural spat collection. Therefore predicting the distribution is of great significance to improve the operating efficiency of the collecting. Hydrodynamic model of Northern China Yellow Sea was established and the motions equations of physical oceanography and verified by the tidal harmonic constants and the measured data velocities of Dalian Bay. According to the passivity drift characteristics of the larvae, combined with the hydrodynamic model and the particle tracking model, the spatial and temporal distribution prediction model was established and the spatial and temporal distribution of the larvae under the influence of flow and wind were simulated. It can be concluded from the model results: ocean currents have greatest impacts on the passive drift path and diffusion of Patinopecten yessoensis larvae; the impact of wind is also important, which changed the direction and speed of the drift. Patinopecten yessoensis larvae were generated in the sea along Zhangzi Island and Guanglu-Dachangshan Island, but after two months, with the impact of wind and currents, the larvae appeared in the west of Dalian and the southern of Lvshun, and even in Bohai Bay. The model results are consistent with the relevant literature on qualitative analysis, and this conclusion explains where the larvae come from in the perspective of numerical simulation.

Keywords: numerical simulation, Patinopecten yessoensis larvae, predicting model, spatial and temporal distribution

Procedia PDF Downloads 304

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

Procedia PDF Downloads 567

Authors: Tahammal Hussain, Khuram Zia, Mumammad Jalal Arif, Megha Parajulee, Abdul Hakeem

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The Bioassay on neonate, 2nd and 3rd instar larvae of Helicoverpa armigera (Hubner) were conducted against Bacillus thuringiensis proteins Cry1Ac. Cry1Ac was incorporated into an artificial diet and was serially diluted with distilled water and then mixed with diet at an appropriate temperature of diet. Toxins incorporated prepared diet was poured into Petri-dishes. For controls, distilled water was mixed with the diet. Five toxin doses 0.25, 0.5, 1, 2, and 4 ug / ml and one control were used for each instars of H. armigera 20 larvae were used in each replication and each treatment is replicated four times. LC50 of Cry1Ac against neonate, 2nd and 3rd instar larvae of H. armigera were 0.34, 0.81 and 1.46 ug / ml. So Cry1Ac is more effective against neonate larvae of H .armigera as compared to 2nd and 3rd instar larvae under laboratory conditions.

Keywords: B. thuringiensis, Cry1Ac, H. armigera, toxicity

Procedia PDF Downloads 413

Authors: Zunnu Raen Akhtar, Farhan Ali, Muhammad Saeed-Ur-Rehman

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Jassids are considered as serious sucking pests in eggplants. Jassids can be controlled using imidacloprid, but it can also result in non-target ecological impacts on eco-system. It can also result in reduced population of predators of jassids in the field. An experiment was conducted on jassids, Amrasca sp. reared on eggplant leaves were treated with insecticide imidacloprid at lower, recommended and higher doses including 1L, 2L, 3L respectively. 3rd instar larvae and adults of jassids were exposed to lower, recommended, higher doses. Mortality tests were repeated three times for each dose and insect growth stage. Imidacloprid was sprayed on the leaves followed by drying. Data was recorded for 4, 8, 12, 16, 20, 24 hours after spraying insecticide on the leaves. Results showed that higher mortality was observed in higher and recommended doses, while slow mortality was observed in the case of lower dose. It can be asserted that higher and recommended doses causing immediate mortality of insects are better to control Amrasca sp. in the field, it will not cause immediate resistance development in insects against imidacloprid.

Keywords: Amrasca sp., imidacloprid, egg plant, efficacy

Procedia PDF Downloads 225

Authors: Funda Kul, İsmail Gür

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Pension scheme providers have to price mortality risk by accurate mortality forecasting method. There are many mortality-forecasting methods constructed and used in literature. The Lee-Carter model is the first model to consider stochastic improvement trends in life expectancy. It is still precisely used. Mortality forecasting is done by mortality index in the Lee-Carter model. It is assumed that mortality index fits ARIMA time series model. In this paper, we propose and use dynamic normal inverse gaussian distribution to modeling mortality indes in the Lee-Carter model. Using population mortality data for Italy, France, and Turkey, the model is forecasting capability is investigated, and a comparative analysis with other models is ensured by some well-known benchmarking criterions.

Keywords: mortality, forecasting, lee-carter model, normal inverse gaussian distribution

Procedia PDF Downloads 360

Authors: Irma Andriani, Ita Djuwita, Komar Sumantadinata, Alimuddin

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The recent study has been conducted to develop testicular germ cell transplantation as a tool for preservation and propagation of male germ-plasm from endangered fish species, as well as to produce surrogate broodstock of commercially valuable fish. Giant gourami testis had been used as a model for donor and Nile tilapia larvae as recipient. We developed testicular cell xenotransplantation by optimizing the timing of intraperitoneal cell transplantation to recipient larvae aged 1, 3, 5 and 7 days post hatching (dph). Freshly isolated testis of giant gourami weighing 600–800 g were minced in dissociation medium and then incubated for 3 hours in room temperature to collect monodisperce cell suspension. Donor cells labeled with PKH 26 were transplanted into the peritoneal cavity of Nile tilapia larvae using glass micropipettes. Parameters observed were survival rate of Nile tilapia larvae at 24 hours post transplantation (pt) and colonization efficiency of donor cells at 2 and 3 months pt. The incorporated donor cells were observed under fluorescent microscope. The result showed that the lowest survival rate at 24 hours pt was 1 dph larvae (82.74±6.76%) and the highest survival rate were 3 and 5 dph larvae (95.00±5.00% and 95.00±2.50%, respectively). The highest colonization efficiency was on 3 dph larvae (61.1±34.71%) and the lowest colonization efficiency was on 7 dph larvae (19.43±17.33%). In conclusion, 3 dph Nile tilapia larvae was the best recipient for giant gourami testicular germ cells xenotransplantation.

Keywords: xenotransplantation, testicular germ cell, giant gourami, Nile tilapia, colonization efficiency

Procedia PDF Downloads 582