Chronic toxicity of dietary copper to <i>Daphnia magna</i>

There is a growing concern that dietborne metal toxicity might be important in aquatic ecosystems. However, the science behind this matter is insufficiently developed to explicitly and accurately account for this in metal regulation or risk assessment. We investigated the effects of a chronic exposure of Daphnia magna to an elevated level of Cu (3000 µg Cu/g dry wt) in their diet (the green alga Pseudokirchneriella subcapitata). Compared to daphnids fed with P. subcapitata containing a background of 10.6 µg Cu/g dry wt, daphnids fed for 21 days with this Cu-contaminated food accumulated a total copper body burden of 325 µg Cu/g dry wt, which is about 30-fold higher than the control body burden of 12.1 µg/g dry wt. The exposed daphnids experienced a 38% reduction of growth (measured as final dry body weight), a 50% reduction of reproduction (total number of juveniles produced per daphnid), and only produced three broods versus four broods by the control daphnids. Unlike most other studies, we were able to demonstrate that these effects were most likely not due to a reduced nutritional quality of the food, based on C:P ratios and fatty acid content and composition of the Cu-contaminated algae. Life-history analysis showed that time to first brood was not affected by dietary Cu, while the second and third broods were significantly delayed by 0.7 and 1.5 days, respectively. On the other hand, brood sizes of all three broods were significantly lower in Cu exposed daphnids, i.e. by 32–55%. The variety of effects observed suggest the possible, and perhaps simultaneous, involvement of several toxicity mechanisms such as increased metabolic cost, reduced energy acquisition (potentially via inhibition of digestive enzyme activity), targeted inhibition of reproduction (potentially via inhibition of vitellogenesis), and/or direct inhibition of molting. Further research is needed to differentiate between these postulated mechanisms of dietary Cu toxicity and to determine whether they act separately or in concert

Reproductive toxicity of dietary zinc to <i>Daphnia magna</i>

Regulatory assessments of metals in freshwaters are mostly based on dissolved metal concentrations, assuming that toxicity is caused by waterborne metal only. Little attention has been directed to the toxicity of dietary metals to freshwater invertebrates. In this study the chronic toxicity of dietary zinc to Daphnia magna was investigated. The green alga Pseudokirchneriella subcapitata was exposed for 64 h to a control and three dissolved zinc concentrations, i.e. 23, 28 and 61 µg L-1, resulting in internal zinc burdens in the algae of 130, 200, 320 and 490 µg g-1 dry weight, respectively. These algae were used as a food source in chronic, 21-day bioassays with D. magna in a test medium to which no dissolved zinc was added. None of the treatments resulted in effects on feeding rates or somatic growth of D. magna. In contrast, a significant 40% decrease of total reproduction (number of juveniles per adult) was observed in the 28 and 61 µg L-1 treatments. Time to first brood was not affected, whereas the mean brood size and the fraction of reproducing parent daphnids were reduced from the second brood onwards and the magnitude of these reductions increased with each subsequent brood. The reduced reproduction was accompanied with an elevated zinc accumulation in the 61 µg L-1 treatment only, suggesting that total body burden is no good indicator of dietary zinc toxicity. Overall our data suggest that dietary zinc specifically targets reproduction in D. magna through accumulation in particular target sites, possibly cells or tissues where vitellogenin synthesis or processing occur. Further, our data illustrate that the potential importance of the dietary exposure route should be carefully considered and interpreted in regulatory assessments of zinc

Microbe–microbe interactions trigger Mn(II)-oxidizing gene expression

Manganese (Mn) is an important metal in geochemical cycles. Some microorganisms can oxidize Mn(II) to Mn oxides, which can, in turn, affect the global cycles of other elements by strong sorption and oxidation effects. Microbe-microbe interactions have important roles in a number of biological processes. However, how microbial interactions affect Mn(II) oxidation still remains unknown. Here, we investigated the interactions between two bacteria (Arthrobacter sp. and Sphingopyxis sp.) in a co-culture, which exhibited Mn(II)-oxidizing activity, although neither were able to oxidize Mn(II) in isolation. We demonstrated that the Mn(II)-oxidizing activity in co-culture was most likely induced via contact-dependent interactions. The expressed Mn(II)-oxidizing protein in the co-culture was purified and identified as a bilirubin oxidase belonging to strain Arthrobacter. Full sequencing of the bilirubin oxidase-encoding gene (boxA) was performed. The Mn(II)-oxidizing protein and the transcripts of boxA were detected in the co-culture, but not in either of the isolated cultures. This indicate that boxA was silent in Arthrobacter monoculture, and was activated in response to presence of Sphingopyxis in the co-culture. Further, transcriptomic analysis by RNA-Seq, extracellular superoxide detection and cell density quantification by flow cytometry indicate induction of boxA gene expression in Arthrobacter was co-incident with a stress response triggered by co-cultivation with Sphingopyxis. Our findings suggest the potential roles of microbial physiological responses to stress induced by other microbes in Mn(II) oxidation and extracellular superoxide production