Mosquito larvicidal and ovicidal activity of puffer fish extracts against Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti (Diptera: Culicidae)

The extracts of liver (LE), ovary (OE), skin (SE) and muscle (ME) tissues of four species of puffer fishes viz., Arothron hispidus, Lagocephalus inermis, Lagocephalus scleratus and Chelonodon patoca were evaluated against larvae and eggs of three mosquito vectors, Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti. The LC50 values were 1194.26, 1382.73 (LE); 1421.42, 1982.73 (OE); 7116.86, 15038.98 (ME) and 10817.8 ppm (SE) for An. stephensi and Cx. quinquefasciatus respectively for A. hispidus. In the case of L. inermis, the LC50 values were 1163.83, 1556.1 and 2426.38 (LE); 1653.53, 2734.74 (OE); 6067.47 (ME) and 10283.04 ppm (SE) for An. stephensi, Cx. quinquefasciatus and Ae. aegypti respectively. The LC50 values were 1509.98, 1608.69 (LE) and 1414.9, 2278.69 ppm (OE) for An. stephensi and Cx. quinquefasciatus respectively for the extracts of L. scleratus. In the case C. patoca extracts the LC50 values were 1182.29, 1543.00, 2441.03 (LE) and 1076.13, 2582.11 ppm (OE) for An. stephensi, Cx. quinquefasciatus and Ae. aegypti respectively. OE and LE of all puffer fishes exhibited zero percent egg hatchability from 600 to 1000 ppm against eggs of An. stephensi and Cx. quinquefasciatus. This study shows that puffer toxins are effective in killing the larvae and eggs of mosquitoes.

Development of a controlled release formulation of an indigenous insect growth regulator, DPE-28, a substituted diphenylether, for controlling the breeding of Culex quinquefasciatus.

Background & objectives: DPE-28, a substituted diphenyl ether (2,6-ditertiarybutyl phenyl-2’,4’-dinitro phenyl ether) was reported to exhibit promising insect growth regulating activity against Culex quinquefasciatus, the vector of lymphatic filariasis. A controlled release formulation (CRF) of DPE-28 has been developed to control Cx. quinquefasciatus in its breeding habitats. Toxicity of DPE-28, safety to non-target mosquito predators and the release profile of the CRF of DPE-28 are studied and discussed. Methods: The acute oral and dermal toxicity was tested in male and female Wistar rats as per the Organization for Economic Cooperation and Development (OECD) guidelines 425 and 402 respectively. The toxicity of DPE-28 to non-target predators was tested as per the reported procedure from this laboratory. The CRF of DPE-28 was prepared by following the reported procedure developed at this laboratory earlier. The concentration of DPE-28 released from the CRF was monitored by HPLC by constructing a calibration graph by plotting the peak area in the Y-axis and the concentration of DPE-28 in the X-axis. Results: DPE-28 has been tested for acute oral toxicity and found to be moderately toxic with LD50 value of 1098 mg/kg body weight (b.w). The results of the acute dermal toxicity and skin irritation studies reveal that DPE-28 is safe and non-irritant. DPE-28 when tested at 0.4 mg/litre against non-target mosquito predators did not produce any mortality. The release profile of the active ingredient DPE-28 from the CRF by HPLC technique showed that the average daily release (ADR) of DPE-28 ranged from 0.07 to 5.0 mg/litre during first four weeks. Thereafter the matrix started eroding and the ADR ranged from 5 to 11 mg/litre during the remaining 5 wk. The cumulative release of active ingredient showed that > 90 per cent of the active ingredient was released from the matrix. Interpretation & conclusions: The controlled release matrix of DPE-28 was thus found to inhibit the adult emergence (>80%) of Cx. quinquefasciatus for a period of nine weeks. The CRF of DPE-28 may play a useful role in field and may be recommended for mosquito control programme after evaluating the same under field conditions.

Optimization of media composition for the production of cyclosporin A by Tolypocladium species.

BACKGROUND & OBJECTIVE: Cyclosporins are produced by certain species of the filamentous fungi, belonging to the genus Tolypocladium. While there are numerous reports on the use of cyclosporins in clinical studies, reports on the various aspects of their production have been very limited. Therefore, this study was carried to optimize the medium composition for the production of cyclosporin A, produced by a strain of the filamentous fungus, Tolypocladium species by static fermentation. METHODS: The effect of different nutrients on the production of cyclosporin A, produced by Tolypocladium species in stationary culture was studied by growing the fungus for 21 days at 25 +/- 2 degrees C under different media composition. Cyclosporin A was extracted by homogenizing the fungal cells with methanol and the cyclosporin A level was analyzed by high performance liquid chromatography (HPLC). RESULTS: Among the six different media studied for the production of cyclosporin A, medium 'f' containing glucose (8%), casein acid hydrolysate (3%), malt extract (2%), peptone (1%) and DL- alpha-amino butyric acid (0.5%) favoured the maximum production (2.22 +/- 0.02 g/l medium or 5.85 +/- 0.35 g/kg biomass). INTERPRETATION & CONCLUSION: This study showed that by optimizing the composition of fermentation media enhanced production of cyclosporin A was obtained. Since the strain Tolypocladium (VCRC F21 NRRL No.18950) produces a high level of cyclosporin A in the identified fermentation medium, it could be exploited for industrial production.