M. Sufian


2 Geographic Variation in the Baseline Susceptibility of Helicoverpa armigera (Hubner) (Noctuidae: Lepidoptera) Field Populations to Bacillus thuringiensis Cry Toxins for Resistance Monitoring

Authors: A. Aslam, Muhammad Arshad, Muhammad D. Gogi, M. Sufian


The transgenic cotton expressing Bacillus thuringiensis (Bt) provides an effective control of Helicoverpa armigera, a most damaging pest of the cotton crop. However, Bt cotton may not be the optimal solution owing to the selection pressure of Cry toxins. As Bt cotton express the insecticidal proteins throughout the growing seasons, there are the chances of resistance development in the target pests. A regular monitoring and surveillance of target pest’s baseline susceptibility to Bt Cry toxins is crucial for early detection of any resistance development. The present study was conducted to monitor the changes in the baseline susceptibility of the field population of H. armigera to Bt Cry1Ac toxin. The field-collected larval populations were maintained in the laboratory on artificial diet and F1 generation larvae were used for diet incorporated diagnostic studies. The LC₅₀ and MIC₅₀ were calculated to measure the level of resistance of population as a ratio over susceptible population. The monitoring results indicated a significant difference in the susceptibility (LC₅₀) of H. armigera for first, second, third and fourth instar larval populations sampled from different cotton growing areas over the study period 2016-17. The variations in susceptibility among the tested insects depended on the age of the insect and susceptibility decreased with the age of larvae. The overall results show that the average resistant ratio (RR) of all field-collected populations (FSD, SWL, MLT, BWP and DGK) exposed to Bt toxin Cry1Ac ranged from 3.381-fold to 7.381-fold for 1st instar, 2.370-fold to 3.739-fold for 2nd instar, 1.115-fold to 1.762-fold for 3rd instar and 1.141-fold to 2.504-fold for 4th instar, depicting maximum RR from MLT population, whereas minimum RR for FSD and SWL population. The results regarding moult inhibitory concentration of H. armigera larvae (1-4th instars) exposed to different concentrations of Bt Cry1Ac toxin indicated that among all field populations, overall Multan (MLT) and Bahawalpur (BWP) populations showed higher MIC₅₀ values as compared to Faisalabad (FSD) and Sahiwal (SWL), whereas DG Khan (DGK) population showed an intermediate moult inhibitory concentrations. This information is important for the development of more effective resistance monitoring programs. The development of Bt Cry toxins baseline susceptibility data before the widespread commercial release of transgenic Bt cotton cultivars in Pakistan is important for the development of more effective resistance monitoring programs to identify the resistant H. armigera populations.

Keywords: Bt cotton, baseline, H. armigera, Cry1Ac toxins

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

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


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: Silkworm, Mortality, heavy-metals, larval-instars, lethal-concentration

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