Evaluations of larvicidal activity of medicinal plant extracts to Aedes aegypti （Diptera： Culicidae） and other effects on a non target fish

A preliminary study was conducted to investigate the effects of the extracts of 112 medicinal plant species, collected from the southern part of Thailand, on Aedes aegypti. Studies on larvicidal properties of plant extracts against the fourth instar larvae revealed that extracts of 14 species showed evidence of larvicidal activity. Eight out of the 14 plant species showed 100% mosquito larvae mortality. The LC50 values were less than 100μg/mL （4.1μg/ mL-89.4μg/mL）. Six plant species were comparatively more effective against the fourth instar larvae at very low concentrations. These extracts demonstrated no or very low toxicity to guppy fish （Poecilia reticulata）, which was selected to represent most common non-target organism found in habitats ofAe. aegypti, at concentrations active to mosquito larvae. Three medicinal plants with promising larvicidal activity, having LC50 and LC50 values being 4.1 and 16.4 μg/mL for Mammea siamensis, 20.2 and 34.7 μg/mL forAnethum graveolens and 67.4 and 110.3μg/mL forAnnona muricata, respectively, were used to study the impact of the extracts on the life cycle ofAe. aegypti. These plants affected pupal and adult mortality and also affected the reproductive potential of surviving adults by reducing the number of eggs laid and the percentage of egg hatchability. When each larval stage was treated with successive extracts at the LC50 value, the first instar larvae were found to be very susceptible to A. muricata and the second instar larvae were found to be susceptible to A. graveolens, while the third and fourth instar larvae were found to be susceptible to M. siamensis. These extracts delayed larval development and inhibited adult emergence and had no adverse effects on P. reticulata at LC50 and LC50 values, except for the M. siamensis extract at its LC50 value.     

The Use of Agar Nutrient Solution to Simulate Lack of Convection in Waterlogged Soils

Agar at 0.1% in nutrient solution (‘stagnant solution’)was used to prevent turbulence (convection), thus simulatingthe slow gas movements which occur in waterlogged soils. Wheat,aged between 6 and 16 d at the start of the treatment, was usedto test plant growth and development in this stagnant solutionfor 8–15 d. K-MES buffer at 5 mol m^-3was used to retainthe pH of the rhizosphere in the stagnant solution at pH 6.5. The prevention of convection reduced dissolved oxygen concentrationsin the bulk solution from 0.275 to below 0.05 mol m^-3after 1d, while ethylene accumulated over 10 d to 6.5x10^-6m³m^-3(ppm). Aerenchyma of nodal roots grown in stagnant solution comprised22% of the cross sectional area of the root 50 mm behind theroot tip; this was similar to values recorded earlier for nodalroots of wheat in waterlogged soil and contrasts with 7.6% forroots in non-flushed solution without agar (referred to in thispaper as ‘semi-stagnant solution’) and 2.4% in N₂-flushedsolution. Increases in dry weight and numbers of nodal roots with timewere larger for stagnant and N₂-flushed, than for semi-stagnantor aerated solution. In contrast, seminal roots did not growin stagnant solution, while seminal roots in N₂-flushed solutiongrew much less than in semi-stagnant or aerated solution. In the stagnant solution, relatively high concentrations ofN, K and P were required to avoid limitations in mineral uptakeinto the roots, due to the long diffusion pathway from the bulksolution imposed by the lack of convection. Nevertheless, ourdata show that the slow growth imposed by the lack of convectionwas due to factors other than low mineral nutrition. The mostlikely cause was the change in the dissolved gas compositionof the root media, particularly of the rhizosphere. In conclusion, in terms of anatomy and morphology the rootsgrown in the stagnant solution more closely resembled thosefrom waterlogged soil than did those grown in either semi-stagnantor N₂-flushed solution. Triticum aestivum; wheat; waterlogging; lack of convection; aerenchyma; root development; nutrient uptake     

Optimization of diet and culture environment for larvae and juvenile freshwater pearl mussels,Hyriopsis (Limnoscapha) myersiana Lea, 1856

Summary

Culture of the freshwater pearl mussel, Hyriopsis (Limnoscapha) myersiana, was carried out in three consecutive steps: (1) culture of glochidia larvae in artificial media, (2) rearing the early juveniles (0–120 days old) in a nursery, and (3) rearing the juveniles (120–360 days old) in an earthen pond. The percentage survival of glochidia in standard tissue culture medium (M199) supplemented with common carp plasma was 95±2.5. All surviving larvae (100%) transformed to juveniles, the duration of transformation being 8 days. The early juveniles (0–60 days old) were fed with a mixture of four selected phytoplankton species (Chlorella sp., Kirchneriella incurvata, Navicula sp. and Coccomyxa sp.). The survival rate of juveniles was 8±0.2%. The average length of these juveniles increased from 0.13±0.01 mm to 1.41±0.16 mm and the average height from 0.16±0.01 mm to 0.98±0.09 mm. Subsequently, 60–120-day juveniles were fed with one of the same four phytoplankton species or a combination of the four. Feeding the juveniles with K. incurvata resulted in the highest survival rate (65±8.32%), with an average length of 3.46±0.04 mm and an average height of 1.94±0.04 mm. Finally, the 120–360-day juveniles were cultured in an earthen pond. There were progressive changes in average weight (0.0037±0.002 g to 11.24±5.02 g), length (3.48±0.39 mm to 54.08±6.21 mm), height (1.97±0.24 mm to 25.09±2.48 mm) and width (0.98±0.06 mm to 12.28±3.21 mm) from 120 to 360 days. The average growth rates per day of these parameters were 0.0497±0.01 g, 0.2414±0.15 mm, 0.0975±0.08 mm and 0.0493±0.03 mm, respectively. H. (L.) myersiana juveniles developed the complete structural composition of the adult by 160 days, and at 360 days, gametogenesis was complete.