Assessment of potential mercury toxicity to native invertebrates in a high-gradient stream.

This study evaluated potential effects of mercury (Hg) on benthic macroinvertebrates in the South River, Virginia, USA. The study used a multiple lines of evidence approach, including spatially and temporally matched sediment chemistry, biological, and toxicological information (Sediment Quality Triad), exposure and effect analysis in bulk and interstitial sediment and interstitial water, and critical body residue analysis. Ten-day Chironomus dilutus and Hyalella azteca toxicity tests established site-specific no-effect concentrations (NOEC) at 18.9 µg/g total Hg (THg) and 102 ng/g methylmercury (MeHg). However, the benthic community at these locations was impaired, with lower mayfly and caddisfly composition. Few locations had concentrations of THg and MeHg that exceeded the NOEC in bulk or interstitial sediment. The THg concentrations in interstitial water were far below concentrations expected to reduce survival in benthic invertebrates, and only a low percentage of samples exceeded sublethal (growth) low-effect concentrations (LOEC) for THg or MeHg. The THg concentrations in invertebrate tissue did not exceed the NOEC or LOEC in the South River, and MeHg concentrations exceeded the LOEC at all locations, including those with no evidence of benthic impairment, illustrating the uncertainty associated with this line of evidence. Finally, statistical modeling that evaluated diversity of sensitive invertebrate species as a function of Hg exposure, geomorphological parameters, and physicochemical variables indicated that physicochemical and geomorphological parameters were most predictive of benthic community; where Hg was indicated, we were unable to distinguish between models with or without interstitial water Hg. Overall, the lines of evidence indicate that Hg, while clearly toxic to invertebrates at sufficiently high exposure concentrations, is not negatively impacting invertebrate communities in the South River. This study illustrates the difficulty of assessing risks to invertebrates using traditional tools of risk assessment and identifies critical gaps in knowledge that complicate the management of Hg risk. Integr Environ Assess Manag 2019;00:000-000. © 2019 SETAC.

Contribution of coexisting sulfate and iron reducing bacteria to methylmercury production in freshwater river sediments.

We investigated microbial methylmercury (CH(3)Hg) production in sediments from the South River (SR), VA, an ecosystem contaminated with industrial mercury (Hg). Potential Hg methylation rates in samples collected at nine sites were low in late spring and significantly higher in late summer. Demethylation of (14)CH(3)Hg was dominated by (14)CH(4) production in spring, but switched to producing mostly (14)CO(2) in the summer. Fine-grained sediments originating from the erosion of river banks had the highest CH(3)Hg concentrations and were potential hot spots for both methylation and demethylation activities. Sequencing of 16S rRNA genes of cDNA recovered from sediment RNA extracts indicated that at least three groups of sulfate-reducing bacteria (SRB) and one group of iron-reducing bacteria (IRB), potential Hg methylators, were active in SR sediments. SRB were confirmed as a methylating guild by amendment experiments showing significant sulfate stimulation and molybdate inhibition of methylation in SR sediments. The addition of low levels of amorphous iron(III) oxyhydroxide significantly stimulated methylation rates, suggesting a role for IRB in CH(3)Hg synthesis. Overall, our studies suggest that coexisting SRB and IRB populations in river sediments contribute to Hg methylation, possibly by temporally and spatially separated processes.

Proteolysis of interleukin-2, interferon and immunoglobulin by venoms.

Limited proteolysis by venoms was analysed by the cleaved peptide band(s) in SDS-polyacrylamide gel electrophoresis. The venom from Crotalus atrox degraded interferon, interleukin-2, IgG, IgM, and a crude form of acetyl cholinesterase but had no effect on IgA. Although the venom from Androctonus australis did not exert appreciable proteolysis on any of the immunoglobulins it had potent proteolytic activities against interferon and interleukin-2. The venom from Vespula maculifrons had only a minor proteolytic effect on interferon. The proteolysis by venoms was not effectively inhibited by alpha 1-antitrypsin or a2-macroglobulin. Moreover, no appreciable proteolytic activity was detected in the venoms from Bufo arenarum, Apis mellifera and Heloderma suspectrum.

Inhibitory effect of transfer RNA on protein kinases from baker's yeast and rat skeletal muscle.

Sephadex G-200 gel filtration of DNA cellulose-treated crude extracts of rat skeletal muscle, revealed a broad peak-fraction of tRNA-inhibitory protein kinases (PK) coeluted endogenous substrates. In comparison, the elution profile of baker's yeast exhibited multiple peak-fractions of tRNA-inhibiting PK. Various tRNA all showed inhibition to PK. In the presence of regulatory subunit of cyclic AMP-dependent protein kinase, tRNA did not exert synergetic inhibition on PK. Moreover, the interaction of tRNA with active muscle PK fractions could not be monitored by the increment of absorbance at 340 nm. tRNA had no significant regulatory effect on the phosphorylation of actin and myosin.

A further study on the regulation of microbial proteases.

Various agents were tested for their effects on microbial proteases, which activity was monitored by the analysis of cleaved peptide bands in SDS-polyacrylamide gel electrophoresis. Using casein as a substrate, fungal protease (type XIX) was inhibited by the phenyl methyl sulphonyl fluoride, chymostatin, antipain and leupeptin, while bacterial protease (type XXVI) was inhibited by phosphatidyl glycerol, phosphatidyl inositol and sphingosine. MS2 RNA exerted minor inhibition on the bacterial proteolysis of regulatory subunits of cyclic AMP-dependent protein kinase (A-PK). The cleavage of DNA binding protein by both proteases was inhibited, in the presence of MS2 RNA and lambda DNA. In comparison, phosphatidyl serine slightly stimulated the fungal protease on the cleavage of ribonuclease T1. RNA polymerase is a good substrate of the bacterial protease as indicated by the generation of multiple cleaved peptide fragments, whereas alkaline phosphatase is not susceptible to proteolysis.