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USGS2020

Ambiguities in using telomere length for age determination in two North American bat species

The age of an animal, determined by time (chronological age) as well as genetic and environmental factors (biological age), influences the likelihood of mortality and reproduction and thus the animal’s contribution to population growth. For many long-lived species, such as bats, a lack of external and morphological indicators has made determining age a challenge, leading researchers to examine genetic markers of age for application to demographic studies. One widely studied biomarker of age is telomere length, which has been related both to chronological and biological age across taxa, but only recently has begun to be studied in bats. We assessed telomere length from the DNA of known-age and minimum known-age individuals of two bat species using a quantitative PCR assay. We determined that telomere length was quadratically related to chronological age in big brown bats ( Eptesicus fuscus ), although it had little predictive power for accurate age determination of unknown-age individuals. The relationship was different in little brown bats ( Myotis lucifugus ), where telomere length instead was correlated with biological age, apparently due to infection and wing damage associated with white-nose syndrome. Furthermore, we showed that wing biopsies currently are a better tissue source for studying telomere length in bats than guano and buccal swabs; the results from the latter group were more variable and potentially influenced by storage time. Refinement of collection and assessment methods for different non-lethally collected tissues will be important for longitudinal sampling to better understand telomere dynamics in these long-lived species. Although further work is needed to develop a biomarker capable of determining chronological age in bats, our results suggest that biological age, as reflected in telomere length, may be influenced by extrinsic stressors such as disease.

Journal of Mammalogy↗

USGS2013

The relative contribution of methanotrophs to microbial communities and carbon cycling in soil overlying a coal-bed methane seep

Seepage of coal-bed methane (CBM) through soils is a potential source of atmospheric CH 4 and also a likely source of ancient (i.e. 14 C-dead) carbon to soil microbial communities. Natural abundance 13 C and 14 C compositions of bacterial membrane phospholipid fatty acids (PLFAs) and soil gas CO 2 and CH 4 were used to assess the incorporation of CBM-derived carbon into methanotrophs and other members of the soil microbial community. Concentrations of type I and type II methanotroph PLFA biomarkers (16:1ω8c and 18:1ω8c, respectively) were elevated in CBM-impacted soils compared with a control site. Comparison of PLFA and 16s rDNA data suggested type I and II methanotroph populations were well estimated and overestimated by their PLFA biomarkers, respectively. The δ 13 C values of PLFAs common in type I and II methanotrophs were as negative as −67‰ and consistent with the assimilation of CBM. PLFAs more indicative of nonmethanotrophic bacteria had δ 13 C values that were intermediate indicating assimilation of both plant- and CBM-derived carbon. Δ 14 C values of select PLFAs (−351 to −936‰) indicated similar patterns of CBM assimilation by methanotrophs and nonmethanotrophs and were used to estimate that 35–91% of carbon assimilated by nonmethanotrophs was derived from CBM depending on time of sampling and soil depth.

FEMS Microbiology Ecology↗

USGS2011

Use of fatty acid analysis to determine dispersal of Caspian Terns in the Columbia River Basin, U.S.A.

Lethal control, which has been used to reduce local abundances of animals in conflict with humans or with endangered species, may not achieve management goals if animal movement is not considered. In populations with emigration and immigration, lethal control may induce compensatory immigration, if the source of attraction remains unchanged. Within the Columbia River Basin (Washington, U.S.A.), avian predators forage at dams because dams tend to reduce rates of emigration of juvenile salmonids ( Oncorhynchus spp. ), artificially concentrating these prey. We used differences in fatty acid profiles between Caspian Terns ( Hydroprogne caspia ) at coastal and inland breeding colonies and terns culled by a lethal control program at a mid‐Columbia River dam to infer dispersal patterns. We modeled the rate of loss of fatty acid biomarkers, which are fatty acids that can be traced to a single prey species or groups of species, to infer whether and when terns foraging at dams had emigrated from the coast. Nonmetric multidimensional scaling showed that coastal terns had high levels of C 20 and C 22 monounsaturated fatty acids, whereas fatty acids of inland breeders were high in C18:3n3, C20:4n6, and C22:5n3. Models of the rate of loss of fatty acid showed that approximately 60% of the terns collected at Rock Island Dam were unlikely to have bred successfully at local (inland) sites, suggesting that terns foraging at dams come from an extensive area. Fatty acid biomarkers may provide accurate information about patterns of dispersal in animal populations and may be extremely valuable in cases where populations differ demonstrably in prey base.

Washington↗

USGS1995

Duration of mineralization and fluid-flow history of the Upper Mississippi Valley zinc-lead district

Studies of fluid inclusions in sphalerite and biomarkers from the Upper Mississippi Valley zinc district show homogenization temperatures to be primarily between 90 and 150 ° C, yet show relatively low levels of thermal maturity. We use numerical calculations to simulate fluid and heat flow through fracture-controlled ore zones and heat transfer to the adjacent rocks. Combining a best-fit path through fluid-inclusion data with measured thermal alteration of biomarkers, we calculated the time interval during which mineralizing fluids circulated through the Upper Mississippi Valley district to be on the order of 200 ka. Cambrian and Ordovician aquifers underlying the district, principally the St. Peter and Mt. Simon Sandstones, were the source of the mineralizing fluid. The duration of mineralization thus reflects the fluid-flow history of these regional aquifers.

Geology↗

USGS2015

Association of anatase (TiO2) and microbes: unusual fossilization effect or a potential biosignature?

We combined microbial paleontology and molecular biology methods to study the Eyreville B drill core from the 35.3-Ma-old Chesapeake Bay impact structure,Virginia, USA. The investigated sample is a pyrite vein collected from the 1353.81-1353.89 m depth interval, located within a section of biotite granite. The granite is a pre-impact rock that was disrupted by the impact event. A search for inorganic (mineral) biosignatures revealed the presence of micron-size rod morphologies of anatase (TiO2) embedded in chlorite coatings on pyrite grains. Neither the Acridine Orange microbial probe nor deoxyribonucleic acid (DNA) extraction followed by polymerase chain reaction (PCR) amplifi cation showed the presence of DNA or ribonucleic acid (RNA) at the location of anatase rods, implying the absence of viable cells in the investigated area. A Nile Red microbial probe revealed the presence of lipids in the rods. Because most of the lipids are resistant over geologic time spans, they are good biomarkers, and they are an indicator of biogenicity for these possibly 35-Ma-old microbial fossils. The mineral assemblage suggests that rod morphologies are associated with low-temperature (<100 °C) hydrothermal alteration that involved aqueous fl uids. The temporal constraints on the anatase fossils are still uncertain because pre-impact alteration of the granite and postimpact heating may have provided identical conditions for anatase precipitation and microbial preservation.

Geological Society of America Bulletin↗

USGS2020

Evidence of prevalent heat stress in Yukon River Chinook salmon

Migrating adult Pacific salmon ( Oncorhynchus spp.) are sensitive to warm water (>18 °C), with a range of consequences from decreased spawning success to early mortality. We examined the proportion of Yukon River Chinook salmon ( O. tshawytscha ) exhibiting evidence of heat stress to assess the potential that high temperatures contribute to freshwater adult mortality in a northern Pacific salmon population. Water temperatures greater than 18 °C have occurred almost annually in the Yukon River and correspond with low population abundance since the 1990s. Using gene transcription products and heat shock protein 70 biomarkers validated by field experiment, we identified heat stress in half of Chinook salmon examined (54%, n = 477) across three mainstem locations and three tributaries in 2016–2017. Biomarkers tracked wide variation in water temperature (14–23 °C) within a tributary. The proportion of salmon with heat stress differed between years at four of the six locations, with more prevalent heat stress in the warmer year. This work demonstrates that warming water temperatures are currently affecting northern populations of Pacific salmon.

Alaska↗

USGS2019

Geochemically distinct oil families in the onshore and offshore Santa Maria basins, California

The purpose of this work is to identify genetic affinities among 48 crude oil samples from the onshore and offshore Santa Maria basins. A total of 21 source-related biomarker and stable carbon isotope ratios among the samples were assessed to assure that they were unaffected by secondary processes. Chemometric analysis of these data identifies six oil families with map and stratigraphic distributions that reflect organofacies variations within the Miocene Monterey Formation source rock. The data comprise a training set that was used to create a chemometric decision tree to classify newly collected oil samples. Three onshore families originated from two synclines, which may contain one or more pods of thermally mature source rock. Multiple biomarker parameters indicate that the six oil families achieved early oil window maturity in the range of 0.6%–0.7% equivalent vitrinite reflectance. The offshore oil samples consist of one family from Point Pedernales field and two families from the “B” prospect. Geochemical characteristics of these families indicate origins under differing water column and sediment oxicity and carbonate versus siliceous and detrital input in ‘carbonate,’ ‘marl,’ and ‘shale’ organofacies like those in the lower calcareous–siliceous, carbonaceous marl, and clayey–siliceous members of the Monterey Formation elsewhere in coastal California. The corresponding lithofacies and organofacies appear to be linked to the early–middle Miocene climate optimum and subsequent paleoclimatic cooling after circa 14 Ma, a systematic up-section increase in the stable carbon isotope composition of related oil samples, decreased preservation of calcium carbonate shells from planktic foraminifera and coccoliths, and increased preservation of clay-sized siliceous shells of diatoms and radiolarians. The results show that organofacies within the Monterey source rock are responsible for many of the geochemical differences between the oil families. This paleoclimate–organofacies model for crude oil from the Monterey Formation can be used to enhance future exploration efforts in many areas of coastal California.

California↗

USGS2011

Petroleum hydrocarbons in sediment from the northern Gulf of Mexico shoreline, Texas to Florida

Petroleum hydrocarbons were extracted and analyzed from shoreline sediment collected from the northern Gulf of Mexico (nGOM) coastline that could potentially be impacted by Macondo-1 (M-1) well oil. Sediment was collected before M-1 well oil made significant local landfall and analyzed for baseline conditions by a suite of diagnostic petroleum biomarkers. Oil residue in trace quantities was detected in 45 of 69 samples. With the aid of multivariate statistical analysis, three different oil groups, based on biomarker similarity, were identified that were distributed geographically along the nGOM from Texas to Florida. None of the sediment hydrocarbon extracts correlated with the M-1 well oil extract, however, the similarity of tarballs collected at one site (FL-18) with the M-1 well oil suggests that some oil from the Deepwater Horizon spill may have been transported to this site in the Florida Keys, perhaps by a loop current, before that site was sampled.

Florida;Texas↗

USGS2004

Biomonitoring of Environmental Status and Trends (BEST) Program: Environmental contaminants and their effects on fish in the Rio Grande Basin

We collected, examined, and analyzed 368 fish of seven species from 10 sites in the Rio Grande Basin (RGB) during late 1997 and early 1998. Four sites were National Contaminant Biomonitoring Program (NCBP) stations where organochlorine ad elemental contaminants in fish had been monitored from 1969 and through 1986. The other six were USGS-National Stream Quantity Accounting Network (NASQAN) stations where water quality is monitored. The objectives were to document temporal and geographic trends in the concentrations of accumulative organic and inorganic contaminants in RGB fish and the effects of contaminants on the fish; to continue testing the feasibility of incorporating biomarkers (that is, biochemical, histopathological, and other biological indicators of contaminant exposure or effects) into a monitoring program for large U.S. rivers; and to evaluate the compatibility of monitoring methods based on the analysis of fish with those used to monitor water by NASQAN. Common carp ( Cyprinus carpio ; carp) and black basses ( Micropterus sp.; bass) were the targeted species; together, they represented 77% of the fish collected. Each fish was examined in the field for externally and internally visible gross lesions, selected organs were weighed to compute various ponderal and organosomatic indices, and samples of tissues and fluids were obtained and preserved for analysis of fish health and reproductive biomarkers. Composite samples of whole fish from each station were grouped by species and gender and analyzed by instrumental methods for persistent organic and inorganic contaminants and for dioxin-like activity (TCDD-EQ) using the H4IIE rat hepatoma cell bioassay.

Rio Grande basin↗

USGS2020

High prevalence of biliary neoplasia in white perch Morone americana: Potential roles of bile duct parasites and environmental contaminants

Recent surveys of white perch Morone americana from Chesapeake Bay, USA, revealed a high prevalence of hepatic and biliary lesions, including neoplasia, and bile duct parasites. Here, we describe lesions in the liver and gallbladder and evaluate for statistical associations among lesions, parasites, and biomarkers of chemical exposure in fish from 2 tributaries of Chesapeake Bay. Fish were collected from an estuarine site in the Choptank River (n = 122, ages 3-11), a tributary with extensive agriculture within the watershed, and the Severn River (n = 131, ages 2-16), a tributary with extensive urban development. Passive integrative samplers were deployed at the fish collection site and an upstream, non-tidal site in each river for 30 d. Intrahepatic biliary lesions observed in fish from both rivers included neoplasia (23.3%), dysplasia (16.2%), hyperplasia (46.6%), cholangitis (24.9%), and dilated ducts containing plasmodia of Myxidium sp. (24.9%). Hepatocellular lesions included foci of hepatocellular alteration (FHA, 15.8%) and neoplasia in 4 Severn River fish (2.3%). Age of fish and Myxidium sp. infections were significant risk factors for proliferative and neoplastic biliary lesions, age alone was a risk factor for FHA, and Goussia bayae infections were associated with cholangitis and cholecystitis. Lesion prevalence was higher in fish from the Severn River, which contained higher concentrations of PAHs, organochlorine pesticides, and brominated diphenyl ethers. Metabolite biomarkers indicated higher PAH exposures in Severn River fish. This study suggests Myxidium sp. as a promoter of bile duct tumors, but more data are needed to evaluate the biological effects of environmental contaminants in this species.

Chesapeake Bay↗

USGS2020

Millennial-scale climate and human drivers of environmental change and fire activity in a dry, mixed-conifer forest of northwestern Montana

Warm summer temperatures and longer fire seasons are promoting larger, and in some cases, more fires that are severe in low- and mid-elevation, dry mixed-conifer forests of the Northern Rocky Mountains (NRM). Long-term historical fire conditions and human influence on past fire activity are not well understood for these topographically and biophysically heterogeneous forests. We developed reconstructions of millennial-scale fire activity, vegetation change, and human presence at Black Lake, a small closed-basin lake on the Flathead Indian Reservation in the Mission Valley, Northwestern Montana, United States. Fossil pollen, charcoal, and biomarkers associated with human presence were used to evaluate the interaction between climate variability, fire activity, vegetation change and human activity for the past 7000 years. Comparisons among multiple proxies suggest climate variability acted as the primary control on fire activity and vegetation change from the early Holocene until the late Holocene when records suggest fire activity and climate variability decoupled. Specific biomarkers (5β-stanols including coprostanol and epi-coprostanol) associated with human presence indicate humans were present within the Black Lake watershed for thousands of years, although the inferred intensity of human presence is highly variable. A strong relationship between climate variability and fire activity during the early and mid-Holocene weakens during the last few thousand years, suggesting possible increased influence of humans in mediating fire activity in recent millennia, and/or a shift in the interaction between the distribution and abundance of woody fuel and fire severity. Human-set fires during the cooler and wetter late Holocene may have been aimed at maintaining important cultural resources associated with the heterogeneous mosaic of mixed conifer forests within the Black Lake watershed. The paleoenvironmental reconstruction at Black Lake corroborates archeological records that show humans were present within the Black Lake watershed for over 7000 years. Further research is needed to evaluate the evidence for this continuous presence and the possible role that people played in shaping fire regimes and vegetation within low- to mid-elevation mixed-conifer ecosystems of the NRM.

Montana↗

USGS2021

Juvenile African clawed frogs (Xenopus laevis) express growth, metamorphosis, mortality, gene expression, and metabolic changes when exposed to thiamethoxam and clothianidin

Neonicotinoids (NEO) represent the main class of insecticides currently in use, with thiamethoxam (THX) and clothianidin (CLO) primarily applied agriculturally. With few comprehensive studies having been performed with non-target amphibians, the aim was to investigate potential biomarker responses along an adverse outcome pathway of NEO exposure, whereby data were collected on multiple biological hierarchies. Juvenile African clawed frogs, Xenopus laevis , were exposed to commercial formulations of THX and CLO at high (100 ppm) and low (20 ppm) concentrations of the active ingredient. Mortality, growth, development, liver metabolic enzyme activity, and gene expression endpoints were quantified. Tadpoles ( n > 1000) from NF 47 through tail resorption stage (NF 66) were exposed to NEO or to NEO-free media treatments. Liver cell reductase activity and cytotoxicity were quantified by flow cytometry. Compared to control reference gene expressions, levels of expression for NEO receptor subunits, cell structure, function, and decontamination processes were measured by RT-qPCR by using liver and brain. Mortality in THX high was 21.5% compared to the control (9.1%); the metabolic conversion of THX to CLO may explain these results. The NF 57 control tadpoles were heavier, longer, and more developed than the others. The progression of development from NF 57–66 was reduced by THX low, and weight gain was impaired. Liver reductases were highest in the control (84.1%), with low NEO exhibiting the greatest reductions; the greatest cytotoxicity was seen with THX high. More transcriptional activity was noted in brains than in livers. Results affirm the utility of a study approach that considers multiple complexities in ecotoxicological studies with non-target amphibians, underscoring the need for simultaneously considering NEO concentration-response relationships with both whole-organism and biomarker endpoints.

International Journal of Molecular Sciences↗

USGS2024

Fish health altered by contaminants and low water temperatures compounded by prolonged regional drought in the Lower Colorado River Basin, USA

The goal of this study was to assess health of male Common Carp (carp, Cyprinus carpio ) at four sites with a wide range in environmental organic contaminant (EOC) concentrations and water temperatures in Lake Mead National Recreation Area NV/AZ, US, and the potential influence of regional drought. Histological and reproductive biomarkers were measured in 17–30 carp at four sites and 130 EOCs in water per site were analyzed using passive samplers in 2010. Wide ranges among sites were noted in total EOC concentrations (>10Xs) and water temperature/degree days (10Xs). In 2007/08, total polychlorinated biphenyls (tPCBs) in fish whole bodies from Willow Beach (WB) in the free-flowing Colorado River below Hoover Dam were clearly higher than at the other sites. This was most likely due to longer exposures in colder water (12–14 °C) and fish there having the longest lifespan (up to 54 years) for carp reported in the Colorado River Basin. Calculated estrogenicity in water exceeded long-term, environmentally safe criteria of 0.1–0.4 ng/L by one to three orders of magnitude at all sites except the reference site. Low ecological screening values for four contaminants of emerging concern (CEC) in water were exceeded for one CEC in the reference site, two in WB and Las Vegas Bay and three in the most contaminated site LVW. Fish health biomarkers in WB carp had 25% lower liver glycogen, 10Xs higher testicular pigmented cell aggregates and higher sperm abnormalities than the reference site. Sperm from LVW fish also had significantly higher fragmentation of DNA, lower motility and testis had lower percent of spermatozoa, all of which can impair reproduction. Projections from a 3D water quality model performed for WB showed that EOC concentrations due to prolonged regional drought and reduced water levels could increase as high as 135%. Water temperatures by late 21st century are predicted to rise between 0.7 and 2.1 °C that could increase eutrophication, algal blooms, spread disease and decrease dissolved oxygen over 5%.

Arizona, Nevada↗

USGS2018

Necropsy-based wild fish health assessment

Anthropogenic influences from increased nutrients and chemical contaminants, to habitat alterations and climate change, can have significant effects on fish populations. Adverse effects monitoring, utilizing biomarkers from the organismal to the molecular level, can be used to assess the cumulative effects on fishes and other organisms. Fish health has been used worldwide as an indicator of aquatic ecosystem health. The necropsy-based fish health assessment provides data on visible abnormalities and lesions, parasites, condition and organosomatic indices. These can be compared by site, season and sex, as well as temporally, to document change over time. Severity ratings can be assigned to various observations to calculate a fish health index for more quantitative assessment. A drawback of the necropsy-based assessment is that it is based on visual observations and condition factors, which are not as sensitive as tissue and subcellular biomarkers for sublethal effects. Additionally, it is rarely possible to identify causes or risk factors associated with observed abnormalities. So, for instance a raised lesion or "tumor" on the fins, lips or body surface may be a neoplasm. However, it could also be a response to a parasite, chronic inflammation or hyperplasia of normal cells in response to an irritant. Conversely, neoplasms, certain parasites, other infectious agents and many tissue changes are not visible and so may be underestimated. However, during the necropsy-based assessment, blood (plasma), tissues for histopathology (microscopic pathology), genomics and other molecular analyses, and otoliths for aging can be collected. These downstream analyses, together with geospatial analyses, habitat assessments, water quality and contaminant analyses can all be important in comprehensive ecosystem evaluations.

Journal of Visualized Experiments↗

USGS2025

Avak Creek oil occurrence, North Slope, Alaska: Newly discovered oil seep on Native lands, near village of Utqiagvik

An unknown occurrence of oil was detected near Avak Creek on Native lands on the North Slope of Alaska. Determining the source of oil was imperative for allowing stakeholders (Federal, State, and local government agencies and the landowner, an Alaska Native corporation) to make timely and informed decisions and mount a mitigation response, if required. The regional and local geological framework of the Avak Creek site was constructed using seismic surveys, well data, and basin modeling results, to identify local petroleum systems, map structural geometry and faults, define source rock thermal maturity distributions, and infer likely oil-migration pathways. Molecular hydrocarbon fingerprints (biomarkers, diamondoids, compound-specific isotopes) of the oil were compared to those of local and regional oil seeps, exploration well tests, and produced oils. Biomarker acid distributions characterized the history and extent of petroleum biodegradation. Integrating subsurface and geochemical parameters, the oil is interpreted to be a natural seep generated locally, predominantly from the Brookian Lower Cretaceous Hue Shale/gamma-ray zone, rather than an anthropogenic source of pollution. Results highlight sophisticated analytical technologies used to characterize complex, compositionally altered hydrocarbons. Results also advance our understanding of Brookian source rock distribution, subsurface petroleum migration pathways, and Arctic Alaska petroleum systems.

Alaska↗

USGS2003

Molecular and isotopic tracers used to examine sources of organic matter and its incorporation into the food webs of San Francisco Bay

Multiple indicators (Chl a , C : N ratios, [ δ 13 C]POC, and two classes of lipid biomarker compounds- sterols and phospholipid ester-linked fatty acids) were used to evaluate spatial and temporal variations in the origin of particulate organic matter (POM) in the San Francisco Bay (SFB) estuary. Comparisons were made between the northern and southern subestuaries of SFB, as well as along the salinity gradient of northern SFB. Two sample types were collected-seston, which was used to characterize the bulk POM, and tissues of the suspension-feeding bivalve Potamocorbula amurensis -in order to evaluate the assimilable portion of the POM. Samples were collected around biological and physical events (phytoplankton blooms and freshwater inflow) thought to be the primary mechanisms controlling temporal variability in organic matter sources. Seston samples indicate that phytoplankton sources of POM are important throughout the entire SFB system, with additional inputs of organic matter from bacterial and terrestrial vascular plant sources delivered to the northern region. Analysis of biomarker compounds in P. amurensis tissues indicates that phytoplankton supply a large fraction of the assimilable carbon to clams throughout SFB, although isotopic analysis of clam tissues suggests that the origin of this reactive carbon varies spatially and that freshwater algae are an important source of reactive organic matter to clams living in northern SFB.

California↗

USGS2014

Season and application rates affect vaccine bait consumption by prairie dogs in Colorado and Utah, USA

Plague, a zoonotic disease caused by the bacterium Yersinia pestis , causes high rates of mortality in prairie dogs ( Cynomys spp.). An oral vaccine against plague has been developed for prairie dogs along with a palatable bait to deliver vaccine and a biomarker to track bait consumption. We conducted field trials between September 2009 and September 2012 to develop recommendations for bait distribution to deliver plague vaccine to prairie dogs. The objectives were to evaluate the use of the biomarker, rhodamine B, in field settings to compare bait distribution strategies, to compare uptake of baits distributed at different densities, to assess seasonal effects on bait uptake, and to measure bait uptake by nontarget small mammal species. Rhodamine B effectively marked prairie dogs' whiskers during these field trials. To compare bait distribution strategies, we applied baits around active burrows or along transects at densities of 32, 65, and 130 baits/ha. Distributing baits at active burrows or by transect did not affect uptake by prairie dogs. Distributing baits at rates of ≥65/ha (or ≥1 bait/active burrow) produced optimal uptake, and bait uptake by prairie dogs in the autumn was superior to uptake in the spring. Six other species of small mammals consumed baits during these trials. All four species of tested prairie dogs readily consumed the baits, demonstrating that vaccine uptake will not be an obstacle to plague control via oral vaccination.

Colorado, Utah↗

USGS2011

Secretion of anti-Müllerian hormone in the Florida manatee Trichechus manatus latirostris , with implications for assessing conservation status

Environmental and anthropogenic stressors can affect wildlife populations in a number of ways. For marine mammals (e.g. the Florida manatee Trichechus manatus latirostris ), certain stressors or conservation risk factors have been identified, but sublethal effects have been very difficult to assess using traditional methods. The development of 'biomarkers' allows us to correlate effects, such as impaired reproduction, with possible causes. A recently developed biomarker (anti-Müllerian hormone, AMH) provides an enzyme-linked immunosorbent assay of gonadal function. The study objective was to determine AMH levels in wild manatees. In total, 28 male and 17 female manatee serum samples were assayed. Animal demographics included collection date, body weight (kg) and total length (cm). In certain cases, age of individuals was also known. AMH levels ranged from 160 to 2451.85 ng ml -1 (mean = 844.65 ng ml -1 ) in males and 0.00 to 0.38 ng ml -1 (mean = 0.10 ng ml-1) in females. Linear regression analyses revealed a significant relationship between male AMH levels and body weight (R 2 = 0.452; p < 0.001) and length (R 2 = 0.338; p < 0.001). Due to the small sample size, regression analyses for female AMH and body weight and length were not significant. This represents the first report of AMH detection in a marine mammal. AMH levels in male manatees are the highest of any species observed to date, whereas levels in females are within reported ranges. Further studies will promote improved conservation decision by assessing AMH levels in the manatee as a function of various stressors including, but not limited to, nutritional status, serious injuries (e.g. watercraft collisions), exposure to biotoxins or contaminants, or disease.

Florida↗