Induction of biochemical stress markers and apoptosis in transgenic Drosophila melanogaster against complex chemical mixtures: role of reactive oxygen species

The study was aimed to investigate the effect of leachates of solid waste from a flashlight battery factory and a pigment plant on 70 kDa heat shock protein (Hsp70) expression, generation of reactive oxygen species (ROS), antioxidant enzymes activities and apoptosis in Drosophila. Third instar larvae of Drosophila melanogaster transgenic for hsp70 (hsp70-lacZ) were fed on diet mixed with leachates of solid wastes (0.05-2.0%, v/v) released from two industrial plants at three different pHs (7.00, 4.93 and 2.88) for 2-48 h. A concentration- and time-dependent significant change in Hsp70 expression, ROS generation, antioxidant enzymes activities and MDA content was observed in the exposed larvae preceding the antioxidant enzymes activities. Mitochondria-mediated, caspase-dependent apoptotic cell death in the larvae exposed to 1.0 and 2.0% leachates of flashlight battery factory was concurrent with a significant regression in Hsp70 expression and a higher ROS generation. A positive correlation drawn between ROS generation and apoptotic markers and a negative correlation between apoptotic markers and Hsp70 expression in these groups indicated the important role of ROS in the leachate-induced cellular damage. Hsp70 along with antioxidant enzymes offered protection to the organisms exposed to all the tested concentrations of the leachates of pigment plant waste and 0.5% leachate of flashlight battery factory in a cooperative manner when ROS generation was less induced. Conversely, higher levels of ROS generation in the organisms treated with 1.0 and 2.0% leachate of flashlight battery factory after 24 and 48 h resulted in regression of Hsp70 expression in them leading to cell death. The study suggests that (1) leachates of flashlight battery factory waste more adversely affected the organisms in comparison to the leachates of pigment plant waste. (2) Hsp70 may be used as a biomarker of cellular damage in organisms exposed to leachates. (3) Cell based assays using D. melanogaster as an in vivo model may provide important mechanistic information about the adverse effect of xenobiotics.     

Allelopathic effects of <Emphasis Type="Italic">Ceratiola ericoides</Emphasis> (Empetraceae) on germination and survival of six Florida scrub species

Allelopathic inhibition of germination by Florida scrub plants has been demonstrated in the greenhouse and lab, but not in the field. We studied the allelopathic effects of Florida rosemary (Ceratiola ericoides) roots, leaves, and litter leachates on field germination and three-month survival of six Florida scrub species, three habitat generalists (Lechea deckertii, Palafoxia feayi, and Polygonella robusta), and three rosemary scrub specialists (Hypericum cumulicola, Lechea cernua, and Polygonella basiramia). We used AIC and model averaging to evaluate support for a series of non-exclusive hypotheses. Species varied in germination (2.7–24.6%) and survival (39.2–71%) percentages, and in their sensitivity to leachates. Germination of scrub species was most negatively affected by leaf > root > litter leachates, although not all species followed the overall trend. Additional germination suppression by leachate combinations (relative to single leachates) was minimal. Sites did not vary in germination, but seedling survival did differ among sites. This study further documents the negative impact of Florida rosemary leachates on the germination of co-occurring plant species. Allelopathy may be partly responsible for bare sand gaps in Florida rosemary scrub, and therefore be one of the forces structuring Florida rosemary scrub ecosystems.     

Toxicity of landfill leachate to sea urchin development with a focus on ammonia

Sea urchin gametes and embryos serve as a model system to evaluate toxicity in the marine environment. In this study, the toxicity of complex chemical mixtures in leachate samples to sea urchin development was examined with a focus on ammonia, which was the main contaminant of concern in most samples. Two rapid tests, the submitochondrial particle function and bacterial luminescence tests, were also used. Ammonia is highly toxic to sea urchin embryos with an EC50 of 1.3 mg l⁻¹ for the embryos of the Australian sea urchin Heliocidaris tuberculata. Leachate ammonia levels were well above these EC50 concentrations. To assess the contribution of ammonia to leachate toxicity in sea urchin development, we compared the predicted toxic units (PTU) and observed toxic units (OTU) for ammonia for each sample. The PTU/OTU comparison revealed that the sensitivity of the sea urchin embryos to ammonia were altered (enhanced or decreased) by other chemicals in the leachates. This result emphasises the need for parallel chemical analyses and a suite bioassays for evaluating the toxicity of complex and variable chemical mixtures.