Deepwater Horizon oil spill: assessment of potential impacts on the deep soft-bottom benthos; interim data summary report

A study was initiated in May 2011, under the direction of the Deepwater Horizon (DWH) Natural Resource Damage Assessment (NRDA) Deepwater Benthic Communities Technical Working Group (NRDA Deep Benthic TWG), to assess potential impacts of the DWH oil spill on sediments and resident benthic fauna in deepwater (> 200 meters) areas of the Gulf. Key objectives of the study were to complete the analysis of samples from 65 priority stations sampled in September-October 2010 on two DWH Response cruises (Gyre and Ocean Veritas) and from 38 long-term monitoring sites (including a subset of 35 of the original 65) sampled on a follow-up NRDA cruise in May-June 2011. The present progress report provides a brief summary of results from the initial processing of samples from fall 2010 priority sites (plus three additional historical sites). Data on key macrofaunal, meiofaunal, and abiotic environmental variables are presented for each of these samples and additional maps are included to depict spatial patterns in these variables throughout the study region. The near-field zone within about 3 km of the wellhead, where many of the stations showed evidence of impaired benthic condition (e.g. low taxa richness, high nematode/harpacticoid-copepod ratios), also is an area that contained some of the highest concentrations of total petroleum hydrocarbons (TPH), total polycyclic aromatic hydrocarbons (total PAHs), and barium in sediments (as possible indicators of DWH discharges). There were similar co-occurrences at other sites outside this zone, especially to the southwest of the wellhead out to about 15 km. However, there also were exceptions to this pattern, for example at several farther-field sites in deeper-slope and canyon locations where there was low benthic species richness but no evidence of exposure to DWH discharges. Such cases are consistent with historical patterns of benthic distributions in relation to natural controlling factors such as depth, position within canyons, and availability of organic matter derived from surface-water primary production.

Ecological condition of coastal ocean waters of the northwestern Gulf of Mexico: 2011

In August 2011, the NOAA National Ocean Service (NOS) conducted an assessment of the status of ecological condition of soft-bottom habitat and overlying waters of the continental shelf in the northwestern Gulf of Mexico (GOM). The original sampling design included 50 randomly selected sites from the Mississippi River delta to the U.S./Mexican border, representing a total area of 111,162 square kilometers; however, vessel failures and inclement weather precluded sampling at 16 sites in the western-most part of the study region. Sampling was completed at the remaining 34 sites in offshore waters between the Mississippi River delta and Freeport, Texas, representing an estimated 75,591 square kilometers. Field sampling followed standard methods and indicators applied in prior NOAA coastal studies and EPA’s Environmental Monitoring and Assessment Program (EMAP) and National Coastal Assessment (NCA). A key feature adopted from these studies was the incorporation of a random probabilistic sampling design. Such a design provides a basis for making unbiased statistical estimates of the spatial extent of ecological condition relative to various measured indicators and corresponding thresholds of concern. Indicators included multiple measures of water quality, sediment quality, and biological condition (benthic fauna, fish tissue contaminant levels). Water depths ranged from 13 – 83 m throughout the study area. About 9 % of the area had sediments composed of sands (< 20 % silt+clay), 47 % of the area was composed of intermediate muddy sands (20 – 80 % silt+clay), and 44 % of the sampled area consisted of mud (> 80 % silt+clay). About 50 % of the area (represented by 17 sites) had sediment total organic carbon (TOC) concentrations < 5 mg/g and all of the sites sampled had levels of TOC < 20 mg/g, well below the range associated with potentially harmful effects to benthic fauna (> 50 mg/g). Surface salinities ranged from 23.4 – 36.5 psu, with salinity generally increasing with distance west of the Mississippi River delta. Bottom salinities varied between 31.1 and 36.5 psu, with lowest values occurring at shallow, inner-shelf stations. Surface-water temperatures varied between 29.8 and 31.5 ºC, while near-bottom waters ranged in temperature from 19.4 – 31 ºC. An index of density stratification (Δσt) indicated that portions of coastal shelf waters in the northwestern GOM at the time of this sampling were strongly stratified. Values of Δσt at 19 of the 34 sites sampled in this study (56 % of the study area) ranged from 2.2 to 12.4, which is within the range considered to be indicative of strong vertical stratification (Δσt > 2). Stratification was strongest close to the Mississippi River delta, and decreased with distance west of the delta.

Deep-sea benthic footprint of the Deepwater Horizon blowout

The Deepwater Horizon (DWH) accident in the northern Gulf of Mexico occurred on April 20, 2010 at a water depth of 1525 meters, and a deep-sea plume was detected within one month. Oil contacted and persisted in parts of the bottom of the deep-sea in the Gulf of Mexico. As part of the response to the accident, monitoring cruises were deployed in fall 2010 to measure potential impacts on the two main soft-bottom benthic invertebrate groups: macrofauna and meiofauna. Sediment was collected using a multicorer so that samples for chemical, physical and biological analyses could be taken simultaneously and analyzed using multivariate methods. The footprint of the oil spill was identified by creating a new variable with principal components analysis where the first factor was indicative of the oil spill impacts and this new variable mapped in a geographic information system to identify the area of the oil spill footprint. The most severe relative reduction of faunal abundance and diversity extended to 3 km from the wellhead in all directions covering an area about 24 km2. Moderate impacts were observed up to 17 km towards the southwest and 8.5 km towards the northeast of the wellhead, covering an area 148 km2. Benthic effects were correlated to total petroleum hydrocarbon, polycyclic aromatic hydrocarbons and barium concentrations, and distance to the wellhead; but not distance to hydrocarbon seeps. Thus, benthic effects are more likely due to the oil spill, and not natural hydrocarbon seepage. Recovery rates in the deep sea are likely to be slow, on the order of decades or longer.

Ecological condition of coastal ocean waters of the U.S. continental shelf off South Florida: 2007

A study was initiated with field work in May 2007 to assess the status of ecological condition and stressor impacts throughout the U.S. continental shelf off South Florida, focusing on soft-bottom habitats, and to provide this information as a baseline for evaluating future changes due to natural or human-induced disturbances. The boundaries of the study region extended from Anclote Key on the western coast of Florida to West Palm Beach on the eastern coast of Florida, inclusive of the Florida Keys National Marine Sanctuary (FKNMS), and from navigable depths along the shoreline seaward to the shelf break (~100m). The study incorporated standard methods and indicators applied in previous national coastal monitoring programs — U.S. Environmental Protection Agency’s (EPA) Environmental Monitoring and Assessment Program (EMAP) and National Coastal Assessment (NCA) — including multiple measures of water quality, sediment quality, and biological condition. Synoptic sampling of the various indicators provided an integrative weight-of-evidence approach to assessing condition at each station and a basis for examining potential associations between presence of stressors and biological responses. A probabilistic sampling design, which included 50 stations distributed randomly throughout the region, was used to provide a basis for estimating the spatial extent of condition relative to the various measured indicators and corresponding assessment endpoints (where available). The study was conducted through a large cooperative effort by National Oceanic and Atmospheric Administration (NOAA)/National Centers for Coastal Ocean Science (NCCOS), EPA, U.S. Geological Survey (USGS), NOAA/Oceanic and Atmospheric Research (OAR)/Atlantic Oceanographic and Meteorological Laboratory in Miami, FKNMS, and the Florida Fish and Wildlife Conservation Commission (FWC). The majority of the South Florida shelf had high levels of dissolved oxygen (DO) in near-bottom water (> 5 mg L-1) indicative of “good” water quality.. DO levels in bottom waters exceeded this upper threshold at 98.8% throughout the coastal-ocean survey area. Only 1.2% of the region had moderate DO levels (2-5 mg/L) and no part of the survey area had DO <2.0 mg/L. In addition, offshore waters throughout the region had relatively low levels of total suspended solids (TSS), nutrients, and chlorophyll a indicative of oligotrophic conditions. Results suggested good sediment quality as well. Sediments throughout the region, which ranged from sands to intermediate muddy sands, had low levels of total organic carbon (TOC) below bioeffect guidelines for benthic organisms. Chemical contaminants in sediments were also mostly at low, background levels. For example, none of the stations had chemicals in excess of corresponding Effects-Range Median (ERM) probable bioeffect values or more than one chemical in excess of lower-threshold Effects-Range Low (ERL) values. Cadmium was the only chemical that occurred at moderate concentrations between corresponding ERL and ERM values. Sixty fish samples from 28 stations were collected and analyzed for chemical contaminants. Eleven of these samples (39% of sites) had moderate levels of contaminants, between lower and upper non-cancer human-health thresholds, and ten (36% of sites) had high levels of contaminants above the upper threshold.

Ecological Condition of Coastal Ocean Waters within Stellwagen Bank National Marine Sanctuary: 2008

In June 2008, the NOAA National Ocean Service (NOS), in conjunction with the EPA National Health and Environmental Effects Laboratory (NHEERL), conducted an assessment of the status of ecological condition of soft-bottom habitat and overlying waters within the boundaries of Stellwagen Bank National Marine Sanctuary (SBNMS). The sanctuary lies approximately 20 nautical miles east of Boston, MA in the southwest Gulf of Maine between Cape Ann and Cape Cod and encompassing 638 square nautical miles (2,181 km2). A total of 30 stations were targeted for sampling using standard methods and indicators applied in prior NOAA coastal studies and EPA’s Environmental Monitoring and Assessment Program (EMAP) and National Coastal Assessment (NCA). A key feature adopted from these studies was the incorporation of a random probabilistic sampling design. Such a design provides a basis for making unbiased statistical estimates of the spatial extent of ecological condition relative to various measured indicators and corresponding thresholds of concern. Indicators included multiple measures of water quality, sediment quality, and biological condition (benthic fauna, fish tissue contaminant levels). Depths ranged from 31 – 137 m throughout the study area. About 76 % of the area had sediments composed of sands (< 20 % silt-clay), 17 % of the area was composed of intermediate muddy sands (20 – 80 % silt-clay), and 7 % of the sampled area consisted of mud (> 80 % siltclay). About 70 % of the area (represented by 21 sites) had sediment total organic carbon (TOC) concentrations < 5 mg/g and all but one site (located in Stellwagen Basin) had levels of TOC < 20 mg/g, which is well below the range potentially harmful to benthic fauna (> 50 mg/g). Surface salinities ranged from 30.6 – 31.5 psu, with the majority of the study region (approximately 80 % of the area) having surface salinities between 30.8 and 31.4 psu. Bottom salinities varied between 32.1 and 32.5 psu, with bottom salinities at all sites having values above the range of surface salinities. Surface-water temperatures varied between 12.1 and 16.8 ºC, while near-bottom waters ranged in temperature from 4.4 – 6.2 ºC. An index of density stratification (Δσt) indicated that the waters of SBNMS were stratified at the time of sampling. Values of Δσt at 29 of the 30 sites sampled in this study (96.7 % of the study area) varied from 2.1 – 3.2, which is within the range considered to be indicative of strong vertical stratification (Δσt > 2) and typical of the western Gulf of Maine in summer. Levels of dissolved oxygen (DO) were confined to a fairly narrow range in surface (8.8 – 10.4 mg/L) and bottom (8.5 – 9.6 mg/L) waters throughout the survey area. These levels are within the range considered indicative of good water quality (> 5 mg/L) with respect to DO. None of these waters had DO at low levels (< 2 mg/L) potentially harmful to benthic fauna and fish.

Cruise report: regional assessment of ecosystem condition and stressor impacts along the northwestern Gulf of Mexico Shelf. NOAA Ship 'Nancy Foster' NF-11-07-RACOW (August 8-16, 2011)

This cruise report is a summary of a field survey conducted along a portion of the U.S. continental shelf in northwestern Gulf of Mexico (GOM), at navigable depths along the coastline seaward to the shelf break (~100m) from about 89°30' W to 95°28' W longitude, August 8 – 16, 2011 on NOAA Ship Nancy Foster Cruise NF-11-07-RACOW. Synoptic sampling of multiple ecological indicators was conducted at each of 34 stations throughout these waters using a random probabilistic sampling design. The original study design consisted of 50 stations extending from the Mississippi delta all the way to the U.S./Mexican border, but vessel failures precluded sampling at 16 stations within the western-most portion of the study area. At each station samples were collected for the analysis of benthic community structure and composition; concentrations of chemical contaminants in sediments and target demersal biota; sediment toxicity; nutrient and chlorophyll levels in the water column; and other basic habitat characteristics such as salinity, temperature, dissolved oxygen, turbidity, pH, sediment grain size, and organic carbon content. Other indicators, from a human-dimension perspective, were also recorded, including presence of vessels, oil rigs, surface trash, visual oil sheens in sediments or water, marine mammals, or noxious/oily sediment odors. The overall purpose of the survey was to collect data to assess the status of ecosystem condition and potential stressor impacts throughout the region, based on these various indicators and corresponding management thresholds, and to provide this information as a baseline for determining how such conditions may be changing with time. While sample analysis is still ongoing, some preliminary results and observations are reported here. A final report will be completed once all data have been processed.

Cruise report: regional assessment of ecosystem condition and stressor impacts along the northeastern Gulf of Mexico shelf; NOAA Ship Nancy Foster NF-10-09-RACOW (August 13-21, 2010)

This cruise report is a summary of a field survey conducted along the continental shelf of the northeastern Gulf of Mexico (GOM), encompassing 70,062 square kilometers of productive marine habitats located between the Mississippi Delta and Tampa Bay, August 13–21, 2010 on NOAA Ship Nancy Foster Cruise NF-10-09-RACOW. Synoptic sampling of multiple ecological indicators was conducted at each of 50 stations throughout these waters using a random probabilistic sampling design. At each station samples were collected for the analysis of benthic community structure and composition; concentrations of chemical contaminants (metals, pesticides, TPHs, PAHs, PCBs, PBDEs) in sediments and target demersal biota; sediment toxicity; nutrient and chlorophyll levels in the water column; and other basic habitat characteristics such as depth, salinity, temperature, dissolved oxygen, turbidity, pH, CDOM fluorescence, sediment grain size, and organic carbon content. Discrete water samples were collected just below the sea surface, in addition to any deeper subsurface depths where there was an occurrence of suspicious CDOM fluorescence signals, and analyzed for total BTEX/TPH and carcinogenic PAHs using immunoassay test kits. Other indicators of potential value from a human-dimension perspective were also recorded, including presence of any vessels, oil rigs, surface trash, visual oil sheens in sediments or water, marine mammals, or noxious/oily sediment odors. The overall purpose of the survey was to collect data to assess the status of ecosystem condition and potential stressor impacts throughout the region, based on these various indicators and corresponding management thresholds, and to provide this information as a baseline for determining how such conditions may be changing with time. In addition to the original project goals, both the scientific scope and general location of this project are relevant to addressing potential ecological impacts of the Deepwater Horizon oil spill. While sample analysis is still ongoing, a few preliminary results and observations are reported here. A final report will be completed once all data have been processed.

Development, validation, and utilization of a competitive enzyme-linked immunosorbent assay for the detection of antibodies against Brucella species in marine mammals

A competitive enzyme-linked immunosorbent assay (cELISA) was developed by using a whole-cell antigen from a marine Brucella sp. isolated from a harbor seal (Phoca vitulina). The assay was designed to screen sera from multiple marine mammal species for the presence of antibodies against marine-origin Brucella. Based on comparisons with culture-confirmed cases, specificity and sensitivity for cetacean samples tested were 73% and 100%, respectively. For pinniped samples, specificity and sensitivity values were 77% and 67%, respectively. Hawaiian monk seal (Monachus schauinslandi; n = 28) and bottlenose dolphin (Tursiops truncatus; n = 48) serum samples were tested, and the results were compared with several other assays designed to detect Brucella abortus antibodies. The comparison testing revealed the marine-origin cELISA to be more sensitive than the B. abortus tests by the detection of additional positive serum samples. The newly developed cELISA is an effective serologic method for detection of the presence of antibodies against marine-origin Brucella sp. in marine mammals.

Cruise report: summer 2009 Sapelo Island National Estuarine Research Reserve ecological characterization (June 7 - June 13, 2009)

This cruise report is a summary of a field survey conducted within the Sapelo Island National Estuarine Research Reserve (SINERR), located on the Georgia coastline, June 7 – June 13, 2009. Multiple indicators of ecological condition and human dimensions were sampled synoptically at each of 30 stations throughout SINERR using a random probabilistic sampling design. Samples were collected for the analysis of benthic community structure and composition; concentrations of chemical contaminants (metals, pesticides, PAHs, PCBs, PBDEs) in sediments and target demersal biota; nutrient and chlorophyll levels in the water column; bacterial contaminants in the water column; and other basic habitat characteristics such as depth, salinity, temperature, dissolved oxygen, turbidity, total suspended solids, pH, sediment grain size, and organic carbon content. In addition to the fish samples that were collected for analysis of chemical contaminants relative to human-health consumption limits, other human-dimension indicators were sampled as well including presence or absence of fishing gear, vessels, surface trash, and noxious sediment odors. The overall purpose of the survey was to collect data to assess the status of ecosystem condition and potential stressor impacts throughout SINERR, based on these various indicators and corresponding management thresholds, and to provide this information as a baseline for determining how such conditions may be changing with time. While sample analysis is still ongoing a few preliminary results and observations are reported here. A final report will be completed once all data have been processed. The results will provide a comprehensive weight-of-evidence basis for evaluating current condition (aka a “state-of-the-SINEER environmental report”) and serve as a quantitative benchmark for tracking any future changes due to either natural or human disturbances. Another goal of the study is to demonstrate its utility as a possible model for assessing the status of condition at other NEERS sites using similar and consistent methods to promote system-wide regional and national comparisons.

Lobster trap debris in the Florida Keys National Marine Sanctuary: distribution, abundance, density, and patterns of accumulation

The fishery for spiny lobster Panulirus argus in the Florida Keys National Marine Sanctuary is well chronicled, but little information is available on the prevalence of lost or abandoned lobster traps. In 2007, towed-diver surveys were used to identify and count pieces of trap debris and any other marine debris encountered. Trap debris density (debris incidences/ha) in historic trap-use zones and in representative benthic habitats was estimated. Trap debris was not proportionally distributed with fishing effort. Coral habitats had the greatest density of trap debris despite trap fishers’ reported avoidance of coral reefs while fishing. The accumulation of trap debris on coral emphasizes the role of wind in redistributing traps and trap debris in the sanctuary. We estimated that 85,548 ± 23,387 (mean ± SD) ghost traps and 1,056,127 ± 124,919 nonfishing traps or remnants of traps were present in the study area. Given the large numbers of traps in the fishery and the lack of effective measures for managing and controlling the loss of gear, the generation of trap debris will likely continue in proportion to the number of traps deployed in the fishery. Focused removal of submerged trap debris from especially vulnerable habitats such as reefs and hardbottom, where trap debris density is high, would mitigate key habitat issues but would not address ghost fishing or the cost of lost gear.

Distribution, demography, and discard mortality of crabs caught as bycatch in an experimental pot fishery for toothfish (Dissostichus eleginoides) in the South Atlantic

Between March 2000 and April 2001 two commercial fishing vessels fished for toothfish (Dissostichus eleginoides) off South Georgia using pots. A significant number of lithodid crabs (three species of Paralomis spp.) were caught as bycatch. Paralomis spinosissima occurred in shallow water, generally shallower than 700 m. Paralomis anamerae, not previously reported from this area and therefore representing a considerable southerly extension in the reported geographic range of this species, had an intermediate depth distribution from 400 to 800 m. Paralomis formosa was present in shallow waters but reached much higher catch levels (and, presumably, densities) between 800 and 1400 m. Differences were also noted in depth distribution of the sexes and size of crabs. Depth, soak time, and area were found to significantly influence crab catch rates. Few crabs (3% of P. spinosissima and 7% of P. formosa) were males above the legal size limit and could therefore be retained. All other crabs were discarded. Most crabs (>99% of P. formosa, >97% of P. spinosissima, and >90% of P. anamerae) were lively on arrival on deck and at subsequent discard. Mortality rates estimated from re-immersion experiments indicated that on the vessel where pots were emptied directly onto the factory conveyor belt 78–89% of crabs would survive discarding, whereas on the vessel where crabs were emptied down a vertical chute prior to being sorted, survivorship was 38–58%. Of the three, P. anamerae was the most vulnerable to handling onboard and sub-sequent discarding. Paralomis spinosissima seemed more vulnerable than P. formosa.

Poverty in the fisheries, inductors, causes and intervention

The study is prompted by the poverty that persisted among the fishing communities of lake victoria at time of considerable cash inflow into the fisheries development of fish processing industry. There has been need for understanding of the poverty and what strategies would be most appreciate for it's reduction.This study has attempted to respond to the needby identifying the nature and distribution of the poverty within the fisheries lake victoria,Uganda, the factor responsible for itand the options for poverty reduction intervention. The study examined the global and regional perspectives of poverty and wealth distribution, noting that wide disparities existed between the developed and the developing world and also between the developing countries themselves. A historical review of development policies and strategies revealed that while successive strategies were able to contribute to growth, their achievement towards poverty alleviation were less than satisfactory, hence the need for continually developing new strategies. A background to Uganda’s society and economy is provided, examining the demographic, political, environmental and economic conditions of the country. Uganda’s development strategies are reviewed, highlighting the role of the Poverty Eradication Action Plan, Uganda’s main strategy for implementing the policy of poverty reduction and wealth distribution. At the agricultural sector level, the Plan for the Modernisation of Agriculture has been formulated, followed by the National Fisheries Policy, aimed at providing a policy framework for the management and development of the fisheries. An appropriate definition of poverty was formulated, considered relevant to the situation of Lake Victoria. The dimensions of poverty included inadequate basic necessities, low education and health achievements, a sense of insecurity and exposure to risk. The research methodology was enhanced by the examination of the Lélé Model of the Poverty–Environmental Degradation problem, the World Bank Model of Poverty Causation and the subsequent Lake Victoria Model developed in this study. It has provided a plan for the research, the consideration of criteria and a data collection plan. The data collection instruments included secondary data search, key informant interviews and a sample survey based on a structured questionnaire. The study identified all the four dimensions of poverty in the fisheries, provided poverty profiles with respect to the different activities, groups of people and regions in the fisheries, based on consumption poverty. Among the people identified to be in poverty were the fishing labourers, fishers of Oreochromis niloticus and those operating with non-powered boats. In the post-harvest fisheries, large proportions of processors involved in salting and sun-drying, market stall and bicycle traders were in the poverty category. The ethnic groups most affected included the Samia, Basoga and Bakenye while the Districts of Jinja, Bugiri and Busia had the highest proportions of fishers in the poverty category. With respect to the other dimensions of poverty, the study showed that educational achievement was low within the fishing communities. The health status was poor, due mainly to the prevalence of malaria, diarrhoea, bilharzia and HIV/AIDS. There was a sense of insecurity within certain sections of the fishing community, due to leadership weaknesses within the local as well as the Government institutions. Some community members operated in a state of risk because they were vulnerable to episodes of income, health and education. The causes of poverty in fisheries included weaknesses within the institutional and social environment, limitations in the technology available to the poor, resource degradation and unfavourable economic factors. The recommendations of the study for poverty reduction included strengthening of policies, developing links, improving capacities and increasing resources, to be applied at the levels of Central Government, Local Government and of the community. In view of the achievements of the methodology used on this study, involving reference to the models, it is recommended that future research should build upon this model approach, as it will continue to produce results, especially when attempting to forecast changes relating to interventions.

The sequence and de novo assembly of the giant panda genome

Using next-generation sequencing technology alone, we have successfully generated and assembled a draft sequence of the giant panda genome. The assembled contigs (2.25 gigabases (Gb)) cover approximately 94% of the whole genome, and the remaining gaps (0.05 Gb) seem to contain carnivore-specific repeats and tandem repeats. Comparisons with the dog and human showed that the panda genome has a lower divergence rate. The assessment of panda genes potentially underlying some of its unique traits indicated that its bamboo diet might be more dependent on its gut microbiome than its own genetic composition. We also identified more than 2.7 million heterozygous single nucleotide polymorphisms in the diploid genome. Our data and analyses provide a foundation for promoting mammalian genetic research, and demonstrate the feasibility for using next-generation sequencing technologies for accurate, cost-effective and rapid de novo assembly of large eukaryotic genomes.

Natural selection on EPAS1 (HIF2 alpha) associated with low hemoglobin concentration in Tibetan highlanders

By impairing both function and survival, the severe reduction in oxygen availability associated with high-altitude environments is likely to act as an agent of natural selection. We used genomic and candidate gene approaches to search for evidence of such genetic selection. First, a genome-wide allelic differentiation scan (GWADS) comparing indigenous highlanders of the Tibetan Plateau (3,200 3,500 m) with closely related lowland Han revealed a genome-wide significant divergence across eight SNPs located near EPAS1. This gene encodes the transcription factor HIF2 alpha, which stimulates production of red blood cells and thus increases the concentration of hemoglobin in blood. Second, in a separate cohort of Tibetans residing at 4,200 m, we identified 31 EPAS1 SNPs in high linkage disequilibrium that correlated significantly with hemoglobin concentration. The sex-adjusted hemoglobin concentration was, on average, 0.8 g/dL lower in the major allele homozygotes compared with the heterozygotes. These findings were replicated in a third cohort of Tibetans residing at 4,300 m. The alleles associating with lower hemoglobin concentrations were correlated with the signal from the GWADS study and were observed at greatly elevated frequencies in the Tibetan cohorts compared with the Han. High hemoglobin concentrations are a cardinal feature of chronic mountain sickness offering one plausible mechanism for selection. Alternatively, as EPAS1 is pleiotropic in its effects, selection may have operated on some other aspect of the phenotype. Whichever of these explanations is correct, the evidence for genetic selection at the EPAS1 locus from the GWADS study is supported by the replicated studies associating function with the allelic variants.