Sumedha Roy

Potential toxicity of flubendiamide in Drosophila melanogaster and associated structural alterations of its compound eye

Flubendiamide, a comparatively new insecticide, is used against lepidopteran insect pests and presumed safe for non-target Drosophila melanogaster (D. melanogaster). In this study, treatment concentrations (0.5, 1, 2 5, 10, and 20 μg/ml) of flubendiamide, lower than agricultural concentrations (rice 50μg/ml or cotton 100 μg/ml), were able to inhibit acetylcholinesterase activity in third instar larvae of D. melanogaster indicating a neurotoxic potential. In addition, larvae exposed to flubendiamide also manifested increased amounts of stress protein hsp70. The larvae expressing such stress response when allowed to emerge as adults displayed severe eye structure deformities found by scanning electron microscopy. These findings indicate a toxic potential for flubendiamide in D. melanogaster.

Study of the changes in life cycle parameters of Drosophila melanogaster exposed to fluorinated insecticide, cryolite

The study explored variations in the life cycle parameters in Drosophila melanogaster as a function of treatment with fluorinated insecticide, cryolite. Some of the life cycle parameters considered in this study were larval duration, pupal duration, and percentage of adult fly emergence in D. melanogaster of Oregon R strain. Freshly hatched first instar larvae were transferred to different dietary concentrations of the test chemical (5, 10, 15, 20, 40, 60, 80, 100, 150, and 200 ppm). Larval duration, pupal duration, and the emergence of flies of both treated and control groups were recorded. Results obtained show a significant (p < 0.001–0.05) change in the mentioned parameters in the treated flies when compared with the controls. Interestingly, the percentage emergence of flies shows a decreasing trend along with increase in treatment concentration and almost no detectable emergence is observed in 200 ppm treatment category until the 20th day of experiment. Thus, the study indicates insecticide-induced variation in duration of different life stages and thereby suggests an effect of the fluorinated insecticide on the biology of a nontarget organism like Drosophila.

Effect of acute exposure of acephate on hemocyte abundance in a non-target victim Drosophila melanogaster

Organophosphate insecticide acephate was tested to investigate effects on hemocyte abundance in a non-target dipteran insect Drosophila melanogaster. For this purpose, third-instar larvae were fed on four graded concentrations (2, 4, 6, or 8 μg/ml) acephate for a period of 12 and 24 hr (acute treatment). Control groups were simultaneously maintained for comparison. Relative proportions of plasmatocytes in hemolymph smear were found to fall with increasing concentrations of the test chemical. Similar decreasing trend in population of lamellocytes was also noted after 12 and 24 hr chemical exposure. In contrast to plasmatocytes and lamellocytes, crystal cell number was found to rise with increasing pesticide concentration. Several factors like oxidative stress, apoptosis induction, and mitotic failure might be the cause of reduced plasmatocyte and lamellocyte count. The elevated number of crystal cells in hemolymph smears is directly indicative of high melanin synthesis that assists larvae to combat chemical stress, since melanin is well known for its potential to minimize physical, chemical, and pathogenic stress.

Acephate-induced shortening of developmental duration and early adult emergence in a nontarget insect Drosophila melanogaster

Similar to several environmental monitoring studies, the present study used Drosophila melanogaster as a model nontarget organism to explore the interfering effects of an organophosphate (OP) insecticide acephate on insect life cycle parameters. Acephate, a common OP, is readily available in nature from agricultural sources as an environmental contaminant. Along with target pests, nontarget fruit flies also suffer exposure to such environmental chemical. To evaluate the effects of such exposure, initially, acute LC50 of acephate for third instar larvae was investigated and found to be between 14 and 16 μg/ml. This information yielded the following experimental concentrations (0.5, 1.5, 2.5, 3.5, 4.5, and 5.5 μg/ml) of test chemical for evaluation of effect, if any, on the insect model. Results showed that mean larval duration of insect significantly decreased on treatment with acephate, whereas the mean pupal duration remained unaffected. Interestingly the decreasing trend was seen to persist in case of mean adult emergence, where treated flies emerged significantly earlier in comparison to controls. Thus, the study demonstrated that acephate-induced shortening of developmental time and early emergence in Drosophila melanogaster.

Exploring hazards of acute exposure of Acephate in Drosophila melanogaster and search for L-ascorbic acid mediated defense in it

This study reveals protective role of l-ascorbic acid (25, 50 and 100μg/mL) against toxic impacts of acute sub-lethal exposure of Acephate (5μg/mL) in a non-target organism Drosophila melanogaster. Organismal effect was evident from increased impairment in climbing activities (9 folds) of treated individuals who also manifested altered ocular architecture. These anomalies were reduced with l-ascorbic acid (l-AA) supplementation. Acephate induced apoptotic lesions in eye imaginal discs and gut confirmed tissue damage that also reduced with l-AA co-treatment. Reduction in viability of fat body cells (∼41%), neural cells (∼42%) and hemocytes (3 folds) indicates cytotoxic and immunotoxic potential of Acephate, which were significantly mitigated with l-AA co-administration. The sub-cellular toxic impacts of Acephate treatment became obvious from enhancement in activities of antioxidant enzymes (CAT by ∼1.63 folds, SOD by ∼1.32 folds), detoxifying enzymes (Cyp450 by ∼1.99 folds and GST by ∼1.34 folds), 2.1 times boost in HSP 70 expression, and inhibition of cholinesterase activity (by ∼0.66 folds). DNA breaks evident through comet assay confirmed Acephate triggered genotoxicity which could also be prevented through co-administration of. L-AA Furthermore, the study proposes the use of Drosophila as a model to screen chemicals for their protective potential against pesticide toxicity.

Structural alterations in compound eye of Drosophila melanogaster in response to sodium fluoride treatment

Sodium fluoride, used as a pesticide in agriculture, is also an ingredient of toothpastes which help to fight dental problems. In this study, Drosophila melanogaster, a non-target organism, is used to explore morphological changes in the adult compound eye as a function of exposure to fluoride at 20, 40, 80, 100 and 150 mg L−1, using scanning electron microscopy. The experimental concentrations were much lower than the ones used in the studies of the British Association of Community Dentistry. Distinct morphological alterations in the eye of the treated insects revealed ommatidial ridges and disoriented mechanosensory bristles which were most prominent in the 40 and 80 mg L−1 treatment groups. Since humans and Drosophila share homology in many genes that are involved in developmental pathways, the present findings raise concern on the use of sodium fluoride as a pesticide.

Exposure-dependent variation in cryolite induced lethality in the nontarget insect, Drosophila melanogaster

Abstract The starting point of toxicity testing of any chemical in an organism is the determination of its Lethal Concentration 50 (LC50). In the present study, LC50 of a fluorinated insecticide cryolite is determined in a non-target insect model, Drosophila melanogaster. Interestingly, the result shows that acute LC50 of cryolite was much greater in comparison to the chronic one in case of Drosophila larvae. Larvae which were exposed to 65,000 to 70,000 μg/ml cryolite through food showed 50% mortality after 18 hours of acute exposure, whereas only 150 to 160 μg/ml cryolite was sufficient to cause 50% mortality in case of chronic exposure. Thus cryolite in a small amount when applied once cannot produce noticeable changes in Drosophila, whereas the same amount when used continuously can be fatal. The non-feeding pupal stage was also seen to be affected by chemical treatment. This suggests that the test chemical affects the developmental fate and results in failure of adult emergence. Absence of chemical-induced mortality in adults assumes that the toxicity of cryolite might be restricted to the preimaginal stages of the organism. Reduction in body size of larvae after ingestion of cryolite (with food) in acute treatment schedule is another interesting finding of this study. Some individuals consuming cryolite containing food cannot survive whereas the few survivors manifest a significant growth retardation which might be due to a tendency of refusal in feeding. Hence the present findings provide a scope of assessment of risk of other similar non-target groups

Effect of Thiovit® Jet on the structure of thoracic microtrichia/trichomes in Drosophila melanogaster

Widely used fungicides and pesticides are known to have profound effect on several nontarget organisms, which is a cause of concern. The present study aims to demonstrate the effect of a fungicide, Thiovit® Jet on the structure of epidermal microtrichia (trichome) of the dorsal thorax in Drosophila melanogaster. External morphology and structural variations of thoracic appendages have been extensively studied using scanning electron microscope from flies treated with different concentrations of Thiovit Jet (20, 30, 40 or 200 μg/ml). Similar to the effect of other fungicides like captan and captafol which are reported to produce somatic mutations in the same organism, the present study successfully demonstrates variation in the trichome/microtrichia structure of the dorsal thorax of D. melanogaster. Structural variations were observed to be associated with different concentrations of Thiovit Jet (30, 40 and 200 μg/ml), but the maximum notable change was found with 40 μg/ml treatment. The gross abnormality in the trichome structure may be due to mutation in proteins associated with normal cuticular deposition.

Trans-generational transmission of altered phenotype resulting from flubendiamide-induced changes in apoptosis in larval imaginal discs of Drosophila melanogaster

The eye and wing morphology of Drosophila melanogaster maintain unique, stable pattern of genesis from larval eye and wing imaginal discs. Increased apoptosis in cells of eye and wing discs was found to be associated with flubendiamide (fluoride containing insecticide) exposure (at the range 0.25-10μg/mL) in D. melanogaster larvae. The chemical fed larvae on attaining adulthood revealed alterations in morphology and symmetry of their compound eyes and wings through scanning electron microscopy. Nearly 40% and 30% of flies (P generation) demonstrated alterations in eyes and wings respectively. Transmission electron microscopic study (at the range 1-20μg/mL) also established variation in the rhabdomere and pigment cell orientation as well as in the shape of the ommatidium. Subsequent SEM study with F1 and F2 generation flies also revealed structural variation in eye and wing. Decrease in percentage of altered eye and wing phenotype was noted in subsequent generations (P> F1>F2). Thus, the diamide insecticide, flubendiamide, expected to be environmentally safe at sub-lethal concentrations was found to increase apoptosis in larvae and thereby cause morphological alteration in the adult D. melanogaster. This study further demonstrated trans-generational transmission of altered phenotype in three subsequent generations of a non-target insect model, D. melanogaster.

Altered differential hemocyte count in 3rd instar larvae of Drosophila melanogaster as a response to chronic exposure of Acephate.

Abstract Acephate, an organophosphate (OP) pesticide, was used to investigate the effects of its chronic exposure on hemocyte abundance in a non-target dipteran insect Drosophila melanogaster. For this purpose, six graded concentrations ranging from 1 to 6 μg/ml were selected, which are below the reported residual values (up to 14 μg/ml) of the chemical. 1st instar larvae were fed with these concentrations up to the 3rd instar stage and accordingly hemolymph smears from these larvae were prepared for differential hemocyte count. Three types of cells are found in Drosophila hemolymph, namely, plasmatocytes, lamellocytes and crystal cells. Plasmatocyte count was found to decrease with successive increase in treatment concentrations. Crystal cells showed an increasing trend in their number. Though the number of lamellocytes was very low, a bimodal response was noticed. Lamellocyte number was found to increase with the initial three concentrations, followed by a dose dependent reduction in their number. As hemocytes are directly linked to the immune system of fruit flies, fluctuations in normal titer of these cells may affect insect immunity. Hemocytes share homologies in their origin and mode of action with the immune cells of higher organisms including man. Thus the present findings suggest that immune cells of humans and other organisms may be affected adversely under chronic exposure to Acephate.