Formation of Brominated Organic Compounds and Molecular Transformations in Dissolved Organic Matter (DOM) after Ballast Water Treatment with Sodium Dichloroisocyanurate Dihydrate (DICD).

Estuarine water treated with a ballast water management system (BWMS) using a solution of dissolved dichloroisocyanurate dihydrate (DICD) resulted in the formation of newly described brominated disinfection byproducts (Br-DBPs). Analysis of dissolved organic matter (DOM) in untreated water with ultrahigh resolution Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) identified 3897 m/z ions and their exact molecular formulas. After DICD treatment, a total of 213 halogenated molecular ions with relative abundance of at least 1% were assigned and confirmed using isotope simulation. Halogenated ions were assigned in four DBP elemental groups including CHOBr (180), CHONBr (13), CHOCl (16), and CHOBrCl (4). Forty-nine of the 197 brominated formulas have not been previously reported. We also were able to tentatively assign possible structures to the formula C3HBr3N2 due to very limited isomeric possibilities. The tentatively assigned compound found at 6.4% relative abundance was identified as either tribromoimidazole or tribromopyrazole. Our results show the formation of complex halogenated DBPs that are formed in the treatment of water with a novel BWMS that employs granular DICD as a biocide. The toxicological and mutagenic properties as well as the fate of these newly identified brominated DBPs are unknown.

In Vitro and In Vivo Assessment of Aqueously Extractable Estrogens in Poultry Manure after Pilot-scale Composting.

Poultry manure contains free and conjugated forms of the natural estrogens 17ß-estradiol and estrone, which can be transported to receiving waters via runoff when land-applied. Previous studies have demonstrated estrogens in runoff from poultry manure-amended fields but have not tracked changes in estrogenicity within this water over time. Microbial conversion of conjugated estrogens (a major portion of water-extractable estrogens) to parent forms may result in temporary increases in estrogenicity in natural water bodies. The present study created 80-L batches of simulated poultry manure runoff, which were investigated over 10 d for estrogenicity by bioluminescent yeast estrogen screen assay and fathead minnow () vitellogenin induction model. The efficacy of different compost conditions (in-vessel aeration ± turning, and piling) on reduction/elimination of aqueously extractable estrogens in poultry manure was also investigated. Results indicate 3- to 10-fold increases in estrogenicity in various poultry manure mixtures during 10-d observations. Estrogenicity returned to low levels in postcompost treatments but remained elevated in the precompost treatment. Aerated compost resulted in >75% reductions in initial, peak, and 10-d mean estrogenicity in aqueous mixtures (0.3, 0.8, and 0.5 ng 17ß-estradiol equivalents [EEQ] L, respectively) compared with the precompost mixture (1.4, 4.8, and 2.1 ng EEQ L, respectively). Estrogenicity was significantly higher in the aqueous extract from the piled treatment than the aerated treatment, and 10-d exposure of male fish to the piled treatment resulted in statistically significant vitellogenin induction. Collectively, our results suggest a need to investigate estrogenicity in surface waters for several days after receiving manure-influenced runoff.

Environmental concerns of roxarsone in broiler poultry feed and litter in Maryland, USA.

Roxarsone has been used extensively in the broiler chicken industry. We reviewed the environmental concerns of this usage. To summarize, arsenic added to poultry feed as roxarsone ends up in poultry litter. Fresh litter contains predominately roxarsone, whereas aged litter contains predominately inorganic arsenic. Soil arsenic concentrations from long-term poultry litter applications can exceed Maryland arsenic soil background remediation standards. Due to continued soil accumulation, arsenic-amended litter use as fertilizer is thought to be unsustainable. Surface-applied roxarsone-amended litter does not influence deep aquifer arsenic concentrations but is transported as inorganic arsenic to receiving waters and very shallow groundwater after precipitation. Arsenic in some receiving waters and sediments from agriculturally dominated watersheds have levels above established criteria. Arsenic in fish and shellfish is mostly organic. Phosphorus-based nutrient management will tend to limit PL application rates in areas that have over-applied phosphorus relative to crop needs, resulting in decreased rates of arsenic application and accumulation. Despite most arsenic in surface soils being tightly bound, as surface soils become more enriched in arsenic, the potential for downward movement increases but is limited in most soils due to the high capacity for binding of arsenic to clay minerals and oxides of iron and aluminum in subsoil horizons. In 2012, Maryland passed a law banning the use of arsenic additives except nitarsone in poultry feed. In 2013, the USFDA withdrew approval of roxarsone, carbarsone, and arsanilic but is reviewing nitarsone.

Using the Sediment Quality Triad to characterize baseline conditions in the Anacostia River, Washington, DC, USA.

The Sediment Quality Triad (SQT) consists of complementary measures of sediment chemistry, benthic community structure, and sediment toxicity. We applied the SQT at 20 stations in the tidal portion of the Anacostia River from Bladensburg, MD to Washington, DC to establish a baseline of conditions to evaluate the effects of management actions. Sediment toxicity was assessed using 10-day survival and growth tests with the freshwater amphipod, Hyalella azteca and the midge, Chironomus dilutus. Triplicate grabs were taken at each station for benthic community analysis and the Benthic Index of Biotic Integrity (B-IBI) was used to interpret the data. Only one station, #92, exhibited toxicity related to sediment contamination. Sediments from this station significantly inhibited growth of both test species, had the highest concentrations of contaminants, and had a degraded benthic community, indicated by a B-IBI of less than 3. Additional sediment from this station was tested and sediment toxicity identification evaluation (TIE) procedures tentatively characterized organic compounds as the cause of toxicity. Overall, forty percent of the stations were classified as degraded by the B-IBI. However, qualitative and quantitative comparisons with sediment quality benchmarks indicated no clear relationship between benthic community health and contaminant concentrations. This study provides a baseline for assessing the effectiveness of management actions in the Anacostia River.

Long-term algal toxicity of oxidant treated ballast water.

National and international regulations require that ships' ballast water is treated to minimize the risk of introducing potentially invasive species. A common approach employed by commercial ballast water management systems is chlorination. This study presents the algal toxicity findings for three chlorination-based BWMS and their implications to environmental safety of port waters receiving treated ballast water from ships. Discharged treated ballast water from all three BWMS was toxic to algae with IC25s (25% growth inhibition) ranging from 9.9% to 17.9%, despite having total residual oxidant concentrations below 0.02 mg/l, based on Whole Effluent Toxicity assays. When held at 4 °C, some of the ballast water samples continued to exhibit toxic effects with no observed effect concentrations as low as 18% after a 134 day holding time. Thirteen individual disinfection by-products were measured above the detected limit at the time of discharge. No correlation between DBPs and algal toxicity was observed.

Same Content, Different Methods: Comparing Lecture, Engaged Classroom, and Simulation.

BACKGROUND AND OBJECTIVES: There is a push to use classroom technology and active teaching methods to replace didactic lectures as the most prevalent format for resident education. This multisite collaborative cohort study involving nine residency programs across the United States compared a standard slide-based didactic lecture, a facilitated group discussion via an engaged classroom, and a high-fidelity, hands-on simulation scenario for teaching the topic of acute dyspnea. The primary outcome was knowledge retention at 2 to 4 weeks. METHODS: Each teaching method was assigned to three different residency programs in the collaborative according to local resources. Learning objectives were determined by faculty. Pre- and posttest questions were validated and utilized as a measurement of knowledge retention. Each site administered the pretest, taught the topic of acute dyspnea utilizing their assigned method, and administered a posttest 2 to 4 weeks later. Differences between the groups were compared using paired t-tests. RESULTS: A total of 146 residents completed the posttest, and scores increased from baseline across all groups. The average score increased 6% in the standard lecture group (n=47), 11% in the engaged classroom (n=53), and 9% in the simulation group (n=56). The differences in improvement between engaged classroom and simulation were not statistically significant. CONCLUSIONS: Compared to standard lecture, both engaged classroom and high-fidelity simulation were associated with a statistically significant improvement in knowledge retention. Knowledge retention after engaged classroom and high-fidelity simulation did not significantly differ. More research is necessary to determine if different teaching methods result in different levels of comfort and skill with actual patient care.

Genome Sequence of Gordonia Bacteriophage Lucky10.

Lucky10 is a newly isolated phage of Gordonia terrae 3612 that was recovered from a soil sample in Pittsburgh, PA. Lucky10 has siphoviral morphology and a double-stranded DNA (dsDNA) genome of 42,979 bp, with 70 predicted protein-coding genes. Lucky10 shows little similarity to previously reported Gordonia phages.

Derivation of acute ecological risk criteria for chlorite in freshwater ecosystems.

Chlorine dioxide has been proposed as an alternative to chlorine because it is a powerful disinfection agent that does not react with ammonia or chlorinated organics to form chloramines or trihalomethanes. The major reduction product formed when chlorine dioxide is added to water is the chlorite ion. Chlorite has been shown to be less toxic than free or combined chlorine. However, previous studies with eight freshwater families produced a US Environmental Protection Agency (USEPA) acute water quality criteria (WQC) lower than the WQC for chlorine. In the current study, an additional 12 families were added to the toxicological database to determine ecological risk criteria (ERC) using both the standard USEPA WQC methodology and the USEPA Office of Pesticide Programs (OPP) probabilistic approach. Chlorite LC50 values for the 20 family database ranged from 0.27 mg/L for the Daphnidae to 208.76 mg/L for the Salmonidae. The Daphnidae were by far the most sensitive family tested, with an LC50 44 times lower than the next most sensitive family tested, the Hyalellidae (LC50=1.19 mg/L) and 214 times lower than the most sensitive fish family tested, the Ictaluridae (LC50=5.79 mg/L). The Criteria Maximum Concentration calculated using the USEPA WQC methodology was 0.025 mg/L while the 95% ERC calculated using the USEPA OPP probabilistic methodology was 0.135 mg/L. The USEPA OPP probabilistic approach yields a criterion more consistent with the overall species sensitivity distribution and is not overly skewed by the low Daphnidae LC50 as is the USEPA WQC approach.

A field test and comparison of acute and chronic sediment toxicity tests with the estuarine amphipod Leptocheirus plumulosus in Chesapeake Bay, USA.

A 28-d partial life-cycle test with the estuarine amphipod Leptocheirus plumulosus was developed in response to the need for an assay to mimic chronic exposure to sediment-associated contaminants. To ensure that toxicity tests have environmental relevance, it is essential to evaluate the relationship between laboratory responses and field measures of contamination. Consequently, one objective of the study was to compare the results of the chronic sediment toxicity test with L. plumulosus to gradients of sediment contamination and the in situ benthic community in its native Chesapeake Bay. Chronic tests were conducted by two laboratories, the Army Corps of Engineers Waterways Experiment Station ([WES]; Vicksburg, MS, USA) and the University of Maryland ([UM] College Park, MD, USA) using different feeding regimes, providing the opportunity to evaluate the effect of this variable on response sensitivity. A second objective was to compare the relative sensitivity of acute and chronic tests with L. plumulosus with field-collected sediments. Overall, there was good agreement between the toxicological response of acute and chronic tests with L. plumulosus and field measures of contamination. Survival in the acute test and chronic test conducted by WES was negatively correlated with concentrations of sediment-associated contaminants. Survival in acute exposures was significantly reduced in sediments from 8 of 11 stations. Indigenous L. plumulosus were found only at two of the three stations that did not exhibit acute toxicity. An unexpected finding was the difference in responsiveness of the two chronic tests. Survival in tests conducted by UM and WES was significantly reduced in sediments from 4 and 6 of 11 stations, respectively. No additional sublethal toxicity was detected in the UM chronic test, but the WES test detected reproductive effects at two additional stations. We believe the observed differences were related to the test diet used. Partly as a result of our findings, the recommended diet for the L. plumulosus chronic test was changed in the final methods document.

Intersex (testicular oocytes) in largemouth bass (Micropterus salmoides) on the Delmarva Peninsula, USA.

The authors describe the prevalence and severity of intersex in the form of testicular oocytes in largemouth bass (Micropterus salmoides) collected over a 5-yr period from a variety of surface waters on the Delmarva Peninsula, USA, a region dominated by poultry production and agricultural land use. During a survey from 2005 to 2007 of approximately 200 male specimens representing 6 fish and 2 frog species collected from numerous small-order streams on Delmarva, intersex was observed in only largemouth bass (system-wide prevalence 17%). During 2008 and 2009, testicular oocytes were encountered in male largemouth bass from 6 lakes and 1 large river system, with prevalence ranging from 33% to 88% (weighted arithmetic mean, 57%). The prevalence of testicular oocytes in largemouth bass from Delmarva lakes was comparable to the highest levels reported in a national US Geological Survey reconnaissance of this species, which also occurred in regions of the Atlantic coastal plain with intensive row-crop and animal agriculture. To the authors' knowledge, the present study represents the first report in the peer-reviewed scientific literature of testicular oocytes in fish on the Delmarva Peninsula.

In vivo imaging in an ABSL-3 regional biocontainment laboratory.

The Regional Biocontainment Laboratory (RBL) at the University of Pittsburgh is a state-of-the-art ABSL-3 facility that supports research on highly pathogenic viruses and bacteria. Recent advances in radiologic imaging provide several noninvasive, in vivo imaging modalities that can be used to longitudinally monitor animals following experimental infection or vaccination. The University of Pittsburgh RBL provides digital radiography, bioluminescence imaging, and PET/CT. Operating these platforms in an ABSL-3 poses unique challenges. This review will discuss the development and refinement of these imaging platforms in high containment, emphasizing specific challenges and how they were overcome.

Decontamination and digestion of infectious animal waste using a tissue dissolver in an animal biosafety level 3 facility.

Alkaline hydrolysis-based tissue dissolvers (TDs) are commercially available tools for the digestion and decontamination of infectious animal waste. The authors carried out a series of experiments to verify whether the TD in their facility completely digested animal carcasses and inactivated infectious agents. Using the manufacturer's recommended cycle parameters, the TD inactivated a high concentration of chemically resistant bacterial spores used as a surrogate for the infectious agents in use in the facility. Animal tissues were completely digested into a non-infectious liquid effluent that could be disposed of directly to the sanitary sewer. Reducing the cycle time by 50% still inactivated all spores, although a small amount of tissue remained undigested. The authors recommend that each facility carry out its own experiments to verify the efficacy of a TD before use, given that the design and style of TD as well as the composition of the tissue load is likely to vary between facilities.

Poultry litter-induced endocrine disruption in fathead minnow, sheepshead minnow, and mummichog laboratory exposures.

Animal feeding operations in the United States produce more than 500 million tons of manure annually. Disposal of poultry waste via application as fertilizer results in substantial runoff of poultry litter-associated contaminants (PLAC). Of particular concern are sex steroids, 17ß-estradiol, estrone and testosterone, responsible for sex differentiation and development of reproductive structures. In a series of laboratory assays, mature male and mixed-sex larval/juvenile fish were continuously exposed to environmentally relevant PLAC solutions. Effects on gonads were assessed histologically, and vitellogenin (VTG) induction was measured as a gauge of estrogenicity. Twenty-one-day exposures to laboratory-generated PLAC solutions routinely induced VTG in mature male Pimephales promelas. Vitellogenesis in Fundulus heteroclitus only occurred at the highest tested PLAC concentration, and Cyprinodon variegatus were unresponsive at any tested concentration. All species produced considerable VTG in response to a 17ß-estradiol-positive control. A pronounced feminization was seen in P. promelas when exposed to PLAC as larvae but not when exposed as juveniles. Runoff from a poultry litter-amended field cropped under standard agronomic practices induced significant VTG in male P. promelas. Results indicate that environmentally relevant PLAC concentrations exhibit endocrine activity sufficient to induce VTG production in male fish and possibly affect sex ratios in resident fish populations.

Assessment of sediment contamination, acute toxicity, and population viability of the estuarine amphipod Leptocheirus plumulosus in baltimore harbor, maryland, USA.

In Chesapeake Bay, Maryland, USA, some of the most contaminated sediments are found in the highly industrialized Baltimore Harbor-Patapsco River area. As part of a comprehensive assessment of sediment quality in this system, sediment toxicity was assessed in 10-d acute tests with the estuarine amphipod Leptocheirus plumulosus. Mean amphipod survival was significantly reduced in 7 of the 25 samples tested despite the occurrence of minor experimental artifacts. The most toxic sediments were collected from Bear Creek; other areas exhibiting toxicity included the Inner Harbor and Colgate Creek. Marginal toxicity was observed in samples from Curtis Creek, Lazeretto Point, and Back River. Negative relationships were detected between survival and concentrations of select sediment-associated contaminants, whereas a very strong positive association existed between survival in laboratory exposures and density of L. plumulosus at the test sites. A weight of evidence approach, including correlation analyses, a model of polycyclic aromatic hydrocarbon bioavailability, and comparisons to benchmark sediment levels, was used to tentatively identify classes of contaminants that contributed to the observed toxicity. Analysis of results suggested that toxicity at stations in Bear Creek and Colgate Creek may have been driven by sediment-associated metals, whereas toxicity at stations in the Inner Harbor was likely due to both metal and organic contaminants. The observed relationships among toxicity test results, concentrations of sediment-associated contaminants, and abundance of L. plumulosus at the test sites suggests that acute toxicity tests with this species are indicative of adverse biological effects in the field.

Assessment of the aquatic and terrestrial toxicity of five biodegradable polymers.

Radiofrequency countermeasures (i.e., chaff) may be released by fighter jets during tactical countermeasures training. Chaff cartridges, pistons, and endcaps (i.e., chaff dispenser materials), all currently made of styrene, are also released into the environment. Accumulation of chaff dispenser materials in the environment is a concern of the Department of Defense. The US Navy is exploring the possibility of constructing degradable chaff dispenser components made of biodegradable polymers. Five polymers are being considered. Degradability and toxicity tests are two of several criteria being used to evaluate various available biodegradable options. Dissolution products from four of five polymers being considered were toxic to aquatic organisms with LC50s/LOELs ranging between 1.24 and 731.30 mg total organic concentration/L. Supernatant from dissolving a 90:10 polyester amide/polyvinyl alcohol copolymer in water for 24h inhibited shoot growth of Brassica rappa and Lepidium sativum. Since our results were obtained using fractions of saturated degradable polymer solutions (1 or 10 g/L), we conclude that the tested degradable polymers were of low toxicity to the seven aquatic organisms and two terrestrial plant species used in our assays. However, our characterization of the toxicity of these degradable polymers may not be applicable to all species or environmental situations. Information gained from these studies will be used for making decisions on which polymers should be used in the engineering of environmentally friendly chaff dispenser cartridges, pistons, and endcaps.