Evaluation of Potentially Nonlethal Sampling Methods for Monitoring Mercury Concentrations in Smallmouth Bass (\u3ci\u3eMicropterus dolomieu\u3c/i\u3e)

We evaluated three potentially nonlethal alternatives to fillet sampling for the determination of mercury (Hg) concentrations in smallmouth bass (Micropterus dolomieu). Fish (n = 62, 226–464 mm total length) from six sites in southern Missouri were captured by electrofishing. Blood samples (1 mL) from each fish were obtained by caudal veinipuncture with a heparinized needle and syringe. Biopsy needle (10 mm x 14 gauge; three cuts per fish; 10–20 mg total dry weight) and biopsy punch (7 mm x 5 mm in diameter, one plug per fish, 30–50 mg dry weight) samples were obtained from the area beneath the dorsal fin. Fillet samples were obtained from the opposite side of the fish. All samples were freeze-dried and analyzed for total Hg by combustion amalgamation atomic absorption spectrophotometry. Mean relative standard deviations (RSDs) of triplicate samples were similar for all four methods (2.2–2.4%), but the range of RSDs was greater for blood (0.4–5.5%) than for the muscle methods (1.8–4.0%). Total Hg concentrations in muscle were 0.0200–0.8809 lg/g wet weight; concentrations in plug, needle, and fillet samples from each fish were nearly identical. Blood Hg concentrations were 0.0006–0.0812 lg/mL and were highly correlated with muscle concentrations; linear regressions between log-transformed blood and fillet Hg concentrations were linear and statistically significant (p \u3c 0.01), and explained 91–93% of the total variation. Correlations between fillet Hg concentrations and fish size and age were weak; together they explained \u3c37% of the total variation, and the relations differed among sites. Overall, any of the alternative methods could provide satisfactory estimates of fillet Hg in smallmouth bass; however, both blood and plug sampling with disposable instruments were easier to perform than needle sampling. The biopsy needle was the most difficult to use, especially on smaller fish, and its relative expense necessitates reuse and, consequently, thorough cleaning between fish to prevent cross-contamination

Concentrations of Cadmium, Lead, and Zinc in Fish from Mining-Influenced Waters of Northeastern Oklahoma: Sampling of Blood, Carcass, and Liver for Aquatic Biomonitoring

The Tri-States Mining District (TSMD) of Missouri (MO), Kansas (KS), and Oklahoma (OK), USA, was mined for lead (Pb) and zinc (Zn) for more than a century. Mining ceased more than 30 years ago, but wastes remain widely distributed in the region, and there is evidence of surface- and groundwater contamination in the Spring River- Neosho River (SR-NR) system of northeastern OK. In October 2001, we collected a total of 74 fish from six locations in the SR-NR system that included common carp (Cyprinus carpio), channel- and flathead catfish (Ictalurus punctatus and Pylodictis olivaris), largemouth- and spotted bass (Micropterus salmoides and Micropterus punctulatus), and white crappie (Pomoxis annularis). We obtained additional fish from locations in MO that included three reference sites and one site that served as a ‘‘positive control’’ (heavily contaminated by Pb). Blood, carcass (headed, eviscerated, and scaled) and liver (carp only) samples were analyzed for cadmium (Cd), Pb, and Zn. Our objectives were to assess the degree to which fish from the OK portionof the SR-NR system are contaminated by these elements and to evaluate fish blood sampling for biomonitoring. Concentrations of Cd and Pb in carp and catfish from OK sites were elevated and Pb concentrations of some approached those of the highly contaminated site in MO, but concentrations in bass and crappie were relatively low. For Zn, correlations were weak among concentrations in the three tissues and none of the samples appeared to reflect site contamination. Variability was high for Cd inall three tissues of carp; differences between sites were statistically significant (p \u3c 0.05) only for blood even though mean liver concentrations were at least 100-fold greater than those in blood. Blood concentrations of Cd and Pb were positively correlated (r2 = 0.49 to 0.84) with the concentration of the same element in carp and catfish carcasses or in carp livers, and the corresponding multiple regression models were highly significant (p \u3c 0.001). Our data indicate that potentially nonlethal blood sampling can be useful for monitoring of selected metals in carp, catfish, and perhaps other fishes

Concentrations of Metals in Aquatic Invertebrates from the Ozark National Scenic Riverways, Missouri

This report summarizes the findings of a study conducted as a pilot for part of a park-wide monitoring program being developed for the Ozark National Scenic Riverways (ONSR) of southeastern Missouri. The objective was to evaluate using crayfish (Orconectes spp.) and Asian clam (Corbicula fluminea) for monitoring concentrations of metals associated with lead-zinc mining. Lead-zinc mining presently (2007) occurs near the ONSR and additional mining has been proposed. Three composite samples of each type (crayfish and Asian clam), each comprising ten animals of approximately the same size, were collected during late summer and early fall of 2005 from five sites on the Current River and Jacks Fork within the ONSR and from one site on the Eleven Point River and the Big River, which are outside the ONSR. The Big River has been contaminated by mine tailings from historical leadzinc mining. Samples were analyzed by inductively coupled plasma mass spectrometry for lead, zinc, cadmium, cobalt, and nickel concentrations. All five metals were detected in all samples; concentrations were greatest in samples of both types from the Big River, and lowest in samples from sites within the ONSR. Concentrations of zinc and cadmium typically were greater in Asian clams than in crayfish, but differences were less evident for the other metals. In addition, differences among sites were small for cobalt in Asian clams and for zinc in crayfish, indicating that these metals are internally regulated to some extent. Consequently, both sample types are recommended for monitoring. Concentrations of metals in crayfish and Asian clams were consistent with those reported by other studies and programs that sampled streams in southeast Missouri

Biomonitoring of Lead, Zinc, and Cadmium in Streams Draining Lead-Mining and Non-Mining Areas, Southeast Missouri, USA

We evaluated exposure of aquatic biota to lead (Pb), zinc (Zn), and cadmium (Cd) in streams draining a Pb-mining district in southeast Missouri. Samples of plant biomass (detritus, periphyton, and filamentous algae), invertebrates (snails, crayfish, and riffle benthos), and two taxa of fish were collected from seven sites closest to mining areas (mining sites), four sites further downstream from mining (downstream sites), and eight reference sites in fall 2001. Samples of plant biomass from mining sites had highest metal concentrations, with means 10- to 60- times greater than those for reference sites. Mean metal concentrations in over 90% of samples of plant biomass from mining sites were significantly greater than those from reference sites. Mean concentrations of Pb, Zn, and Cd in most invertebrate samples from mining sites, and mean Pb concentrations in most fish samples from mining sites, were also significantly greater than those from reference sites. Concentrations of all three metals were lower in samples from downstream sites, but several samples of plant biomass from downstream sites had metal concentrations significantly greater than those from reference sites. Analysis of supplemental samples collected in the fall of 2002, a year of above-average stream discharge, had lower Pb concentrations and higher Cd concentrations than samples collected in 2001, near the end of a multi-year drought. Concentrations of Pb measured in fish and invertebrates collected from mining sites during 2001 and 2002 were similar to those measured at nearby sites in the 1970s, during the early years of mining in the Viburnum Trend. Results of this study demonstrate that long-term Pb mining activity in southeast Missouri has resulted in significantly elevated concentrations of Pb, Cd, and Zn in biota of receiving streams, compared to biota of similar streams without direct influence of mining. Our results also demonstrate that metal exposure in the study area differed significantly among sample types, habitats, and years, and that these factors should be carefully considered in the design of biomonitoring studies

Concentrations of Metals in Aquatic Invertebrates from the Ozark National Scenic Riverways, Missouri

This report summarizes the findings of a study conducted as a pilot for part of a park-wide monitoring program being developed for the Ozark National Scenic Riverways (ONSR) of southeastern Missouri. The objective was to evaluate using crayfish (Orconectes spp.) and Asian clam (Corbicula fluminea) for monitoring concentrations of metals associated with lead-zinc mining. Lead-zinc mining presently (2007) occurs near the ONSR and additional mining has been proposed. Three composite samples of each type (crayfish and Asian clam), each comprising ten animals of approximately the same size, were collected during late summer and early fall of 2005 from five sites on the Current River and Jacks Fork within the ONSR and from one site on the Eleven Point River and the Big River, which are outside the ONSR. The Big River has been contaminated by mine tailings from historical leadzinc mining. Samples were analyzed by inductively coupled plasma mass spectrometry for lead, zinc, cadmium, cobalt, and nickel concentrations. All five metals were detected in all samples; concentrations were greatest in samples of both types from the Big River, and lowest in samples from sites within the ONSR. Concentrations of zinc and cadmium typically were greater in Asian clams than in crayfish, but differences were less evident for the other metals. In addition, differences among sites were small for cobalt in Asian clams and for zinc in crayfish, indicating that these metals are internally regulated to some extent. Consequently, both sample types are recommended for monitoring. Concentrations of metals in crayfish and Asian clams were consistent with those reported by other studies and programs that sampled streams in southeast Missouri

Assessment of Elemental Concentrations in Streams of the New Lead Belt in Southeastern Missouri, 2002–05

Concerns about possible effects of lead-mining activities on the water quality of federally protected streams located in southeastern Missouri prompted a suite of multidisciplinary studies to be conducted by the U.S. Geological Survey. As part of this investigation, a series of biological studies were initiated in 2001 for streams in the current mining region and the prospecting area. In this report, results are examined for trace elements and other selected chemical measurements in sediment, surface water, and sediment interstitial (pore) water sampled between 2002 and 2005 in association with these biological studies. Compared to reference sites, fine sediments collected downstream from mining areas were enriched in metals by factors as large as 75 for cadmium, 62 for cobalt, 171 for nickel, 95 for lead, and 150 for zinc. Greatest metal concentrations in sediments collected in 2002 were from sites downstream from mines on Strother Creek, Courtois Creek, and the West Fork Black River. Sediments from sites on Bee Fork, Logan Creek, and Sweetwater Creek also were noticeably enriched in lead. Sediments in Clearwater Lake, at least 75 kilometers downstream from mining activity, had metal concentrations that were 1.5 to 2.1 times greater than sediments in an area of the lake with no upstream mining activity. Longitudinal sampling along three streams in 2004 indicated that sediment metal concentrations decreased considerably a few kilometers downstream from mining activities; however, in Strother Creek some metals were still enriched by a factor of five or more as far as 13 kilometers downstream from the Buick tailings impoundment. Compared with 2002 samples, metals concentrations were dramatically lower in sediments collected in 2004 at an upper West Fork Black River site, presumably because beneficiation operations at the West Fork mill ceased in 2000. Concentrations of metals and sulfate in sediment interstitial (pore) waters generally tracked closely with metal concentrations in sediments. Metals, including cobalt, nickel, lead, and zinc, were elevated substantially in laboratory-produced pore waters of fine sediments collected near mining operations in 2002 and 2004. Passive diffusion samplers (peepers) buried 4 to 6 centimeters deep in riffle-run stream sediments during 2003 and 2005 had much lower pore-water metal concentrations than the laboratory-produced pore waters of fine sediments collected in 2002 and 2004, but each sampling method produced similar patterns among sites. The combined mean concentration of lead in peeper samples from selected sites located downstream from mining activities for six streams was about 10-fold greater than the mean of the reference sites. In most instances, metals concentrations in surface water and peeper water were not greatly different, indicating considerable exchange between the surface water and pore water at the depths and locations where peepers were situated. Passive sampling probes used to assess metal lability in pore waters of selected samples during 2004 sediment toxicity tests indicated that most of the filterable lead in the laboratory-prepared pore water was relatively non-labile, presumably because lead was complexed by organic matter, or was present as colloidal species. In contrast, large percentages of cobalt and nickel in pore water appeared to be labile. Passive integrative samplers deployed in surface water for up to 3 weeks at three sites in July 2005 confirmed the presence of elevated concentrations of labile metals downstream from mining operations on Strother Creek and, to a lesser extent, Bee Fork. These samplers also indicated a considerable increase in metal loadings occurred for a few days at the Strother Creek site, which coincided with moderate increases in stream discharges in the area

Tropical coastal habitats as surrogates of fish community structure, grazing, and fisheries value

Habitat maps are frequently invoked as surrogates of biodiversity to aid the design of networks of marine reserves. Maps are used to maximize habitat heterogeneity in reserves because this is likely to maximize the number of species protected. However, the technique's efficacy is limited by intra-habitat variability in the species present and their abundances. Although communities are expected to vary among patches of the same habitat, this variability is poorly documented and rarely incorporated into reserve planning. To examine intra-habitat variability in coral-reef fishes, we generated a data set from eight tropical coastal habitats and six islands in the Bahamian archipelago using underwater visual censuses. Firstly, we provide further support for habitat heterogeneity as a surrogate of biodiversity as each predefined habitat type supported a distinct assemblage of fishes. Intrahabitat variability in fish community structure at scales of hundreds of kilometers (among islands) was significant in at least 75% of the habitats studied, depending on whether presence/absence, density, or biomass data were used. Intra-habitat variability was positively correlated with the mean number of species in that habitat when density and biomass data were used. Such relationships provide a proxy for the assessment of intra-habitat variability when detailed quantitative data are scarce. Intra-habitat variability was examined in more detail for one habitat (forereefs visually dominated by Montastraea corals). Variability in community structure among islands was driven by small, demersal families (e. g., territorial pomacentrid and labrid fishes). Finally, we examined the ecological and economic significance of intra-habitat variability in fish assemblages on Montastraea reefs by identifying how this variability affects the composition and abundances of fishes in different functional groups, the key ecosystem process of parrotfish grazing, and the ecosystem service of value of commercially important finfish. There were significant differences in a range of functional groups and grazing, but not fisheries value. Variability at the scale of tens of kilometers (among reefs around an island) was less than that among islands. Caribbean marine reserves should be replicated at scales of hundreds of kilometers, particularly for species-rich habitats, to capture important intra-habitat variability in community structure, function, and an ecosystem process

Effect of Depth and Duration of Cooling on Death or Disability at Age 18 Months Among Neonates With Hypoxic-Ischemic Encephalopathy: A Randomized Clinical Trial

Importance Hypothermia for 72 hours at 33.5°C for neonatal hypoxic-ischemic encephalopathy reduces death or disability, but rates continue to be high. Objective To determine if cooling for 120 hours or to a temperature of 32.0°C reduces death or disability at age 18 months in infants with hypoxic-ischemic encephalopathy. Design, Setting, and Participants Randomized 2 × 2 factorial clinical trial in neonates (≥36 weeks’ gestation) with hypoxic-ischemic encephalopathy at 18 US centers in the Eunice Kennedy Shriver National Institute of Child Health and Human Development Neonatal Research Network between October 2010 and January 2016. Interventions A total of 364 neonates were randomly assigned to 4 hypothermia groups: 33.5°C for 72 hours (n = 95), 32.0°C for 72 hours (n = 90), 33.5°C for 120 hours (n = 96), or 32.0°C for 120 hours (n = 83). Main Outcomes and Measures The primary outcome was death or moderate or severe disability at 18 to 22 months of age adjusted for center and level of encephalopathy. Severe disability included any of Bayley Scales of Infant Development III cognitive score less than 70, Gross Motor Function Classification System (GMFCS) level of 3 to 5, or blindness or hearing loss despite amplification. Moderate disability was defined as a cognitive score of 70 to 84 and either GMFCS level 2, active seizures, or hearing with amplification. Results The trial was stopped for safety and futility in November 2013 after 364 of the planned 726 infants were enrolled. Among 347 infants (95%) with primary outcome data (mean age at follow-up, 20.7 [SD, 3.5] months; 42% female), death or disability occurred in 56 of 176 (31.8%) cooled for 72 hours and 54 of 171 (31.6%) cooled for 120 hours (adjusted risk ratio, 0.92 [95% CI, 0.68-1.25]; adjusted absolute risk difference, −1.0% [95% CI, −10.2% to 8.1%]) and in 59 of 185 (31.9%) cooled to 33.5°C and 51 of 162 (31.5%) cooled to 32.0°C (adjusted risk ratio, 0.92 [95% CI, 0.68-1.26]; adjusted absolute risk difference, −3.1% [95% CI, −12.3% to 6.1%]). A significant interaction between longer and deeper cooling was observed (P = .048), with primary outcome rates of 29.3% at 33.5°C for 72 hours, 34.5% at 32.0°C for 72 hours, 34.4% at 33.5°C for 120 hours, and 28.2% at 32.0°C for 120 hours. Conclusions and Relevance Among term neonates with moderate or severe hypoxic-ischemic encephalopathy, cooling for longer than 72 hours, cooling to lower than 33.5°C, or both did not reduce death or moderate or severe disability at 18 months of age. However, the trial may be underpowered, and an interaction was found between longer and deeper cooling. These results support the current regimen of cooling for 72 hours at 33.5°C

The PtdIns 3-Kinase/Akt Pathway Regulates Macrophage-Mediated ADCC against B Cell Lymphoma

Macrophages are important effectors in the clearance of antibody-coated tumor cells. However, the signaling pathways that regulate macrophage-induced ADCC are poorly defined. To understand the regulation of macrophage-mediated ADCC, we used human B cell lymphoma coated with Rituximab as the tumor target and murine macrophages primed with IFNγ as the effectors. Our data demonstrate that the PtdIns 3-kinase/Akt pathway is activated during macrophage-induced ADCC and that the inhibition of PtdIns 3-kinase results in the inhibition of macrophage-mediated cytotoxicity. Interestingly, downstream of PtdIns 3-kinase, expression of constitutively active Akt (Myr-Akt) in macrophages significantly enhanced their ability to mediate ADCC. Further analysis revealed that in this model, macrophage-mediated ADCC is dependent upon the release of nitric oxide (NO). However, the PtdIns 3-kinase/Akt pathway does not appear to regulate NO production. An examination of the role of the PtdIns 3-kinase/Akt pathway in regulating conjugate formation indicated that macrophages treated with an inhibitor of PtdIns 3-kinase fail to polarize the cytoskeleton at the synapse and show a significant reduction in the number of conjugates formed with tumor targets. Further, inhibition of PtdIns 3-kinase also reduced macrophage spreading on Rituximab-coated surfaces. On the other hand, Myr-Akt expressing macrophages displayed a significantly greater ability to form conjugates with tumor cells. Taken together, these findings illustrate that the PtdIns 3-kinase/Akt pathway plays a critical role in macrophage ADCC through its influence on conjugate formation between macrophages and antibody-coated tumor cells