Conserved Microbial Toxicity Responses for Acute and Chronic Silver Nanoparticle Treatments in Wetland Mesocosms.

Most studies of bacterial exposure to environmental contaminants focus on acute treatments; however, the impacts of single, high-dose exposures on microbial communities may not readily be extended to the more likely scenario of chronic, low-dose contaminant exposures. Here, in a year-long, wetland mesocosm experiment, we compared microbial community responses to pulse (single 450 mg dose of silver) and chronic (weekly 8.7 mg doses of silver for 1 year) silver nanoparticle (Ag0 NP) treatments, as well as a chronic treatment of "aged" sulfidized silver nanoparticles (Ag2S NPs). While mesocosms exposed to Ag2S NPs never differed significantly from the controls, both Ag0 NP treatments exhibited reduced microbial diversity and altered community composition; however, the effects differed in timing, duration, and magnitude. Microbial community-level impacts in the acute Ag0 NP treatment were apparent only within the first weeks and then converged on the control mesocosm composition, while chronic exposure effects were observed several months after exposures began, likely due to interactive effects of nanoparticle toxicity and winter environmental conditions. Notably, there was a high level of overlap in the taxa which exhibited significant declines (>10×) in both treatments, suggesting a conserved toxicity response for both pulse and chronic exposures. Thus, this research suggests that complex, but short-term, acute toxicological studies may provide critical, cost-effective insights into identifying microbial taxa sensitive to long-term chronic exposures to Ag NPs.

A survey of crop-derived transgenes in activated and digester sludges in wastewater treatment plants in the United States.

The use of transgenic crops has become increasingly common in the United States over the last several decades. Increasing evidence suggests that DNA may be protected from enzymatic digestion and acid hydrolysis in the digestive tract, suggesting that crop-derived transgenes may enter into wastewater treatment plants (WWTPs) intact. Given the historical use of antibiotic resistance genes as selection markers in transgenic crop development, it is important to consider the fate of these transgenes. Herein we detected and quantified crop-derived transgenes in WWTPs. All viable US WWTP samples were found to contain multiple gene targets (p35, nos, bla and nptII) at significantly higher levels than control samples. Control wastewater samples obtained from France, where transgenic crops are not cultivated, contained significantly fewer copies of the nptII gene than US activated and digester sludges. No significant differences were measured for the bla antibiotic resistance gene (ARG). In addition, a nested PCR (polymerase chain reaction) assay was developed that targeted the bla ARG located in regions flanked by the p35 promoter and nos terminator. Overall this work suggests that transgenic crops may have provided an environmental source of nptII; however, follow-up studies are needed to ascertain the viability of these genes as they exit WWTPs.

Microbial community response to silver nanoparticles and Ag+ in nitrifying activated sludge revealed by ion semiconductor sequencing.

Silver nanoparticles (AgNPs), which are known to act as biocides, are incorporated into medical and consumer products including athletic clothing, stuffed animals, liquid dietary supplements, and more. The increasing use of AgNPs in these products is likely to lead to their entry into both natural and engineered systems, which has the potential to disrupt bacterial processes including those involved in nutrient cycling in wastewater treatment. In the present study, sequencing batch reactors (SBR) mimicking secondary wastewater treatment were operated to determine the effects of AgNPs on the microbial communities contained within activated sludge of wastewater treatment plants (WWTP). SBRs were treated with 0.2 and 2ppm of either gum Arabic (GA)-coated AgNPs, citrate (Ca)-coated AgNPs, or Ag+ as AgNO3. Cell samples were collected and DNA isolated periodically throughout SBR operation. DNA was used for Ion Torrent Next Gen Sequencing of the V3 region of the 16S rDNA gene. Subsequent analyses revealed that the microbial community both shifted and recovered quickly in response to Ag+. Both AgNP treatments resulted in slower initial community shifts than that observed with the Ag+ treatment. GA-AgNPs elicited the longest lasting effect. Additional examination of nitrogen removal bacteria suggested the possibility of an increase in sludge bulking species with increased concentrations of AgNPs in WWTPs. This study supports the hypothesis that Ag+ release from AgNPs is largely coating-dependent and thus a key driver in dictating AgNP toxicity.

Reducing Environmental Toxicity of Silver Nanoparticles through Shape Control.

The use of antibacterial silver nanomaterials in consumer products ranging from textiles to toys has given rise to concerns over their environmental toxicity. These materials, primarily nanoparticles, have been shown to be toxic to a wide range of organisms; thus methods and materials that reduce their environmental toxicity while retaining their useful antibacterial properties can potentially solve this problem. Here we demonstrate that silver nanocubes display a lower toxicity toward the model plant species Lolium multiflorum while showing similar toxicity toward other environmentally relevant and model organisms (Danio rerio and Caenorhabditis elegans) and bacterial species (Esherichia coli, Bacillus cereus, and Pseudomonas aeruginosa) compared to quasi-spherical silver nanoparticles and silver nanowires. More specifically, in the L. multiflorum experiments, the roots of silver nanocube treated plants were 5.3% shorter than the control, while silver nanoparticle treated plant roots were 39.6% shorter than the control. The findings here could assist in the future development of new antibacterial products that cause less environmental toxicity after their intended use.

The in vitro antitumor activity of vitamins C and K3 against ovarian carcinoma.

BACKGROUND: The objective was to evaluate the cytotoxic effect and mechanism of action of vitamins C (VC) and K3 (VK3) on ovarian carcinoma. MATERIALS AND METHODS: Cytotoxicity assays were performed on ovarian cancer cell lines with VC, VK3 or a VC/VK3 combination. FIC index was employed to evaluate synergism. Flow cytometry was accomplished at 90% cytotoxic doses. Light, transmission electron microscopy and DNA isolation were performed. RESULTS: Antitumor activity was exhibited by both VC, VK3 and VC/VK3. VC/VK3 demonstrated synergistic activity. VC/VK3 may induce a G1 block in the cell cycle. Combined vitamin treatment resulted in cells that maintain apparently intact nuclei while extruding pieces of organelle-free cytoplasm. Degradation of chromosomal DNA was observed. CONCLUSION: Cell death (autoschizis) displayed characteristics of both apoptosis and necrosis. The cytotoxic effects observed may enable vitamins C and K3 to play an adjuvant role in the treatment of ovarian cancer.