Effects of phenanthrene on different levels of biological organization in larvae of the sediment-dwelling invertebrate Chironomus sancticaroli (Diptera: Chironomidae).

The hydrocarbon phenanthrene is an organic compound commonly found in the environment. In aquatic ecosystems, it is highly toxic to organisms, although little is known about its effects on sediment-dwelling organisms. The purpose of this study was to evaluate phenanthrene effects on biochemical, histological, and ontogenetic levels in larvae of the sediment-dwelling invertebrate Chironomus sancticaroli at acute and chronic exposure. Lethal concentrations were estimated and toxicity (acute-96 h and chronic- 8 d) tests were performed at phenanthrene concentrations from 0.12 to 1.2 mg L-1. At acute and chronic exposure, we evaluated acetylcholinesterase (AChE), alpha esterase (EST-α), and beta esterase (EST-ß) activities as well as histological alterations. In the assays with chronic exposure, effects on larval development were estimated using antennae length (instar estimative) and body length (growth estimative). The EST-α showed a significantly increased activity after 48 h at acute exposure to high concentrations of phenanthrene, while EST-ß activity was increased after 48 and 72 h at acute exposure at higher concentrations and at 0.12 mg L-1 at chronic exposure. At acute exposure, the midgut showed alterations such as brush border disruption, gastric caeca regression, and lumen area reduction; the fat body showed nuclear alteration in the trophocytes, while the Malpighian tubules showed brush border reduction and the salivary glands were subject to cytoplasm vacuolation. At chronic exposure, the same alterations were observed, in addition to vacuolar coalescence in the trophocytes of the fat body. Regarding larval development, a reduction of body length was observed with increasing phenanthrene concentrations. Similarly, molting was delayed; in the control group, all larvae were in the fourth instar, while at higher phenanthrene concentrations, larvae were predominantly in the third instar. Phenanthrene had toxic effects on this chironomid, indicating risks for natural populations.

Benzo(a)pyrene Exposure Causes Genotoxic and Biochemical Changes in the Midge Larvae of Chironomus sancticaroli Strixino & Strixino (Diptera: Chironomidae).

Benzo(a)pyrene (BaP) is a carcinogenic polycyclic aromatic hydrocarbon, also found in nature due to human activities. BaP adheres to sediments showing toxic effects on benthic organisms, including midge larvae of the family Chironomidae. We tested for toxic effects of benzo(a)pyrene on Chironomus sancticaroli Strixino & Strixino 1981 using biochemical and genotoxic biomarkers, to identify changes in metabolic and antioxidant pathways, besides neurotoxic and DNA damage. Enzyme activity was compared by exposing larvae to four nominal concentrations (0.47, 2.13, 3.41, and 4.73 µg l-1) and DNA damage to two concentrations (0.47 and 4.73 µg l-1), after exposure at 24, 48, 72, and 96 h. BaP caused neurotoxic effect, showing acetylcholinesterase alterations at different treatments. Changes in the biotransformation pathway were detected, with an increased activity of alpha and beta esterase in 48 h and reduction of glutathione-S-transferase activity in all periods at the highest concentrations. Damage to the antioxidant system was observed by the increase of the superoxide dismutase and reduction of the catalase, in 48 h. Genotoxicity was detected by an increased DNA damage at 48 and 72 h. The lowest concentration (0.47 µg l-1), even presenting low mortality, also altered the biochemical parameters of the larvae. Thus, these results indicate that BaP causes metabolic, neurotoxic, and genotoxic effects on C. sancticaroli, even at low concentrations and short-term exposure. BaP can cause damage of immature invertebrates, and the ecological dynamics can be affected, since these organisms have trophic importance in the aquatic environment.

Comparative sodium transport patterns provide clues for understanding salinity and metal responses in aquatic insects.

The importance of insects in freshwater ecosystems has led to their extensive use in ecological monitoring programs. As freshwater systems are increasingly challenged by salinization and metal contamination, it is important to understand fundamental aspects of aquatic insect physiology (e.g., osmoregulatory processes) that contribute to insect responses to these stressors. Here we compared the uptake dynamics of Na as NaCl, NaHCO3 and Na2SO4 in the caddisfly Hydropsyche betteni across a range of Na concentrations (0.06-15.22 mM) encompassing the vast majority of North American freshwater ecosystems. Sulfate as the major anion resulted in decreased Na uptake rates relative to the chloride and bicarbonate salts. A comparison of Na (as NaHCO3) turnover rates in the caddisfly Hydropsyche sparna and the mayfly Maccaffertium sp. revealed different patterns in the 2 species. Both species appeared to tightly regulate their whole body sodium concentrations (at ∼47±1.8 µmol/g wet wt) across a range of Na concentrations (0.06-15.22 mM) over 7 days. However, at the highest Na concentration (15.22 mM), Na uptake rates in H. sparna (419.1 µM Na g(-1) hr(-1) wet wt) appeared close to saturation while Na uptake rates in Maccaffertium sp. were considerably faster (715 g µM Na g(-1) hr(-1) wet wt) and appeared to not be close to saturation. Na efflux studies in H. sparna revealed that loss rates are commensurate with uptake rates and are responsive to changes in water Na concentrations. A comparison of Na uptake rates (at 0.57 mM Na) across 9 species representing 4 major orders (Ephemeroptera, Plecoptera, Trichoptera and Diptera) demonstrated profound physiological differences across species after accounting for the influence of body weight. Faster Na uptake rates were associated with species described as being sensitive to salinization in field studies. The metals silver (Ag) and copper (Cu), known to be antagonistic to Na uptake in other aquatic taxa did not generally exhibit this effect in aquatic insects. Ag only reduced Na uptake at extremely high concentrations, while Cu generally stimulated Na uptake in aquatic insects, rather than suppress it. These results help explain the lack of insect responses to dissolved metal exposures in traditional toxicity testing and highlight the need to better understand fundamental physiological processes in this ecologically important faunal group.