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Fish can get diseases too

Infectious diseases are increasingly recognized as an important component of the ecology of fish in the wild. Many of the viral, bacterial, protozoan and fungal pathogens of fish that were initially discovered in captive fish have their origin among wild populations; however, the impact of disease among these free-ranging stocks has been difficult to study. At the WFRC, combinations of field and laboratory investigations, aided by the tools of molecular biology, have begun to provide information on the ecology of infectious diseases among natural populations of fish in both freshwater and marine ecosystems.

Fact Sheet

Canyons microbiology studies

Off the eastern coast of the United States, several deep canyons cut through the continental shelf, acting like funnels to move sediment from the shelf out to the deep seafloor. Exposed rock outcrops and ledges along the walls of these canyons provide important habitat for deepsea corals and sponges. Although a few scientific expeditions have visited these canyons in the 1970s (Hecker and Blechschmidt 1979, Hecker et al. 1980), their purpose was mainly to map the contours and capture photographs of the bottom using manned submersibles and towed cameras. Our knowledge of the biodiversity in these complex ecosystems is limited; we know little about the macrofauna (e.g., fishes, crabs, sponges, and deepsea corals) and even less about the microbiota. The research described in this report was conducted from 2011 to 2015 as part of the Bureau of Ocean Energy Management (BOEM) study, entitled “Atlantic Deepwater Canyons” study. This work used molecular and microbiological techniques to examine the microbial ecology and diversity associated with Baltimore and Norfolk canyons. Specifically, this work focused on the microbial ecology of four species of octocorals (Acanthogorgia aspera, Anthothela grandiflora, Paramuricea placomus, and Primnoa resedaeformis), the microbial diversity in sediments within and outside the canyons, and a settling plate experiment designed to characterize microbial biofilm formation on a variety of hard substrates.

OCS Study

Has the time come for big science in wildlife health?

The consequences of wildlife emerging diseases are global and profound with increased burden on the public health system, negative impacts on the global economy, declines and extinctions of wildlife species, and subsequent loss of ecological integrity. Examples of health threats to wildlife include Batrachochytrium dendrobatidis , which causes a cutaneous fungal infection of amphibians and is linked to declines of amphibians globally; and the recently discovered Pseudogymnoascus (Geomyces) destructans , the etiologic agent of white nose syndrome which has caused precipitous declines of North American bat species. Of particular concern are the novel pathogens that have emerged as they are particularly devastating and challenging to manage. A big science approach to wildlife health research is needed if we are to make significant and enduring progress in managing these diseases. The advent of new analytical models and bench assays will provide us with the mathematical and molecular tools to identify and anticipate threats to wildlife, and understand the ecology and epidemiology of these diseases. Specifically, new molecular diagnostic techniques have opened up avenues for pathogen discovery, and the application of spatially referenced databases allows for risk assessments that can assist in targeting surveillance. Long-term, systematic collection of data for wildlife health and integration with other datasets is also essential. Multidisciplinary research programs should be expanded to increase our understanding of the drivers of emerging diseases and allow for the development of better disease prevention and management tools, such as vaccines. Finally, we need to create a National Fish and Wildlife Health Network that provides the operational framework (governance, policies, procedures, etc.) by which entities with a stake in wildlife health cooperate and collaborate to achieve optimal outcomes for human, animal, and ecosystem health.

EcoHealth

Microbial ecology of permafrost soils: Populations, processes, and perspectives

Permafrost microbial research has flourished in the past decades, due in part to improvements in sampling and molecular techniques, but also the increased focus on the permafrost greenhouse gas feedback to climate change and other ecological processes in high latitude and alpine permafrost soils. Permafrost microorganisms are adapted to these extreme environments and remain active at low temperatures and when resources are limited. They are also an important component of global elemental cycles as they regulate organic matter turnover and greenhouse gas production, particularly as permafrost thaws. Here we review the permafrost microbiology literature coupled with an exploration of its historical aspects, with a particular focus on a new understanding advanced by molecular biology techniques. We further identify knowledge gaps and ways forward to improve our understanding of microbial contributions to ecosystem biogeochemistry of permafrost-affected systems.

Permafrost and Periglacial Processes

Low genetic variation and evidence of limited dispersal in the regionally important Belize manatee

The Antillean subspecies of the West Indian manatee Trichechus manatus is found throughout Central and South America and the Caribbean. Because of severe hunting pressure during the 17th through 19th centuries, only small populations of the once widespread aquatic mammal remain. Fortunately, protections in Belize reduced hunting in the 1930s and allowed the country's manatee population to become the largest breeding population in the Wider Caribbean. However, increasing and emerging anthropogenic threats such as coastal development, pollution, watercraft collision and net entanglement represent challenges to this ecologically important population. To inform conservation and management decisions, a comprehensive molecular investigation of the genetic diversity, relatedness and population structure of the Belize manatee population was conducted using mitochondrial and microsatellite DNA. Compared with other mammal populations, a low degree of genetic diversity was detected ( H E =0.455; N A =3.4), corresponding to the small population size and long-term exploitation. Manatees from the Belize City Cayes and Southern Lagoon system were genetically different, with microsatellite and mitochondrial F ST values of 0.029 and 0.078, respectively ( P ≤0.05). This, along with the distinct habitats and threats, indicates that separate protection of these two groups would best preserve the region's diversity. The Belize population and Florida subspecies appear to be unrelated with microsatellite and mitochondrial F ST values of 0.141 and 0.63, respectively ( P ≤0.001), supporting the subspecies designations and suggesting low vagility throughout the northern Caribbean habitat. Further monitoring and protection may allow an increase in the Belize manatee genetic diversity and population size. A large and expanding Belize population could potentially assist in the recovery of other threatened or functionally extinct Central American Antillean manatee populations.

Animal Conservation

Correlating sea lamprey density with environmental DNA detections in the lab

Invasive sea lamprey (Petromyzon marinus Linnaeus, 1758) are currently managed by the Great Lakes Fishery Commission in an effort to reduce pest populations below levels that cause ecological damage. One technique to improve stream population assessments could be molecular surveillance in the form of environmental DNA (eDNA) monitoring. We developed and validated four probe-based quantitative polymerase chain reaction (qPCR) assays, then used two probes (cytb, nd1) to determine whether eDNA concentration was correlated with adult and larval sea lamprey density in the lab. We found a strong positive correlation between adult sea lamprey densities of 2, 20, and 200 individuals/2000L and eDNA concentrations in tanks using both assays (cytb, nd1). For larval laboratory tank density trials, eDNA concentrations were generally near our limit of quantification and there was no significant difference in copy numbers detected between larval sea lamprey densities of 1, 5, and 25 individuals/28L. Therefore, we examined detection probability rather than concentration with laboratory tank densities. We observed a trend of increasing detection probabilities with increased larval sea lamprey density that approached significance suggesting that when DNA copy numbers are low, detection rates may be more informative in predicting varying densities of larval sea lamprey. The ability to assess sea lamprey densities from a water sample could be a powerful tool to improve traditional assessment and stream ranking techniques. Further refinement of this method in the field may make eDNA surveillance of sea lamprey a reliable part of stream assessments. Rapid eDNA analysis from many streams may help focus traditional assessment efforts, thereby improving the efficiency of invasive sea lamprey control efforts.

Management of Biological Invasions

Population genetic studies of the sea otter ( Enhydra lutris ): A review and interpretation of available data

C urrent information about the utility of genetic markers for estimating population structuring in sea otters ( Enhydra lutris ) is reviewed. Analyses of spatial population structuring with biochemical and molecular genetic markers are discussed in the context of the species' ecology and history of exploitation. Studies that have used a diversity of genetic markers including allozymes, mitochondrial DNA (mtDNA), and multilocus minisatellites revealed that geographically spearated populations of sea otters are highly differentiated, though little evidence for phylogeographic structuring was suggested. Analyses of population relationships based on mtDNA haplotype frequency distribution suggested that populations can be separated into four major groups: (1) California; (2) Prince William Sound, Alaska; (3) Kodiak Island, Alaska, and islands of the Aleutian archipelago, including the Commander Islands; and (4) the Kuril Islands. Populations from locales separated by large geographic distances often shared haplotypes, suggesting recent common ancestry and some degree of historical gene flow. THe large differences among populations in nuclear and mtDNA gene frequency suggested strong constrains on contemporary gene flow and/or considerable drift in gene frequencies due to population bottlenecks. No evidence for microgeographic structuring was noted. Levels of genetic diversity within populations varied greatly across the species range but were not related to contemporary estimates of population size.

Conference Paper

Use of body mass, footpad length, and wing chord to determine sex in Swainson's Hawks

Many studies of avian ecology require an expedient means to determine sex, and the use of molecular techniques has provided an effective and accurate means to determine sex of raptors in the field ( Sarasola and Negro 2004 , Donohue and Dufty 2006 ). Sometimes investigators need to rely on morphometric measurements to determine sex of monochromatic species such as Swainson's Hawks ( Buteo swainsoni ) because they lack funds or facilities to use molecular techniques or they are analyzing extant data. Discriminant analyses conducted on morphometric measurements have been effective for sexing many raptor species (e.g., Edwards and Kochert 1986 , Bavoux et al. 2006 , Donohue and Dufty 2006 ). Sarasola and Negro (2004) recently reported on the effectiveness of seven morphometric measurements as means to identify sex of Swainson's Hawks on the hawk's austral summer areas in Argentina. They proposed a combination of the length of forearm, wing chord, and tail as an effective means to determine sex, with an overall accuracy of 93%. However, Sarasola and Negro (2004) did not assess the effectiveness of using footpad length (sometimes called toe-pad) to classify sex of these hawks. Footpad length and mass were quite effective in sexing Golden Eagles ( Aquila chrysaetos ), a species with a degree of dimorphism similar to that of Swainson's Hawks ( Snyder and Wiley 1976 ), with an overall accuracy of 98% ( Edwards and Kochert 1986 ). We here report the utility of using footpad length and mass, as well as wing chord length, as a means of sexing Swainson's Hawks on the nesting grounds.

Idaho

Using gene transcription to assess ecological and anthropological stressors in brown bears

Increasingly, population- and ecosystem-level health assessments are performed using sophisticated molecular tools. Advances in molecular technology enable the identification of synergistic effects of multiple stressors on the individual physiology of different species. Brown bears ( Ursus arctos ) are an apex predator; thus, they are ideal candidates for detecting potentially ecosystem-level systemic perturbations using molecular-based tools. We used gene transcription to analyze 130 brown bear samples from three National Parks and Preserves in Alaska. Although the populations we studied are apparently stable in abundance and exist within protected and intact environments, differences in transcript profiles were noted. The most prevalent differences were among locations. The transcript patterns among groups reflect the influence of environmental factors, such as nutritional status, disease, and xenobiotic exposure. However, these profiles also likely represent baselines for each unique environment by which future measures can be made to identify early indication of population-level changes due to, for example, increasing Arctic temperatures. Some of those environmental changes are predicted to be potentially positive for brown bears, but other effects such as the manifestation of disease or indirect effects of oceanic acidification may produce negative impacts.

Alaska

Drift of larval darters (Family Percidae) in the upper Roanoke River basin, USA, characterized using phenotypic and DNA barcoding markers

Larval fish ecology is poorly characterized because sampling is difficult and tools for phenotypically identifying larvae are poorly developed. While DNA barcoding can help address the latter problem, ‘universal’ primers do not work for all fish species. The Roanoke River in the southeastern United States includes seven darters (Family Percide: Tribe Etheostomatini). We made 393 collections of larval fishes in 2015 and 2018, examined darter larvae for morphometric and pigmentation traits, developed PCR primers amplifying darter DNA, and evaluated three gear types for collecting larval darters. Amplified DNA sequences for 1351 larvae matched archived mitochondrial cytochrome oxidase I sequences for darters occurring in the ecosystem. Larval darters were classified to genus with 100% accuracy using the ratio of pectoral fin length to body length; however, identification to species using morphometrics alone was subject to a misclassification rate of 11.8%, which can be resolved by considering pigmentation patterns. Gear-types varied considerably in their capture efficacy for larval darters; most Percina larvae were collected in drift nets. Larval Percina species appeared in the drift before Etheostoma species in both study years. Application of molecular genetic and phenotypic tools to larval fish identification can advance understanding of larval darter ecology.

North Carolina, Virginia

Fluid spatial dynamics of West Nile virus in the USA: Rapid spread in a permissive host environment

The introduction of West Nile virus (WNV) into North America in 1999 is a classical example of viral emergence in a new environment, with its subsequent dispersion across the continent having a major impact on local bird populations. Despite the importance of this epizootic, the pattern, dynamics and determinants of WNV spread in its natural hosts remain uncertain. In particular, it is unclear whether the virus encountered major barriers to transmission, or spread in an unconstrained manner, and if specific viral lineages were favored over others indicative of intrinsic differences in fitness. To address these key questions in WNV evolution and ecology we sequenced the complete genomes of approximately 300 avian isolates sampled across the USA between 2001-2012. Phylogenetic analysis revealed a relatively ‘star-like' tree structure, indicative of explosive viral spread in US, although with some replacement of viral genotypes through time. These data are striking in that viral sequences exhibit relatively limited clustering according to geographic region, particularly for those viruses sampled from birds, and no strong phylogenetic association with well sampled avian species. The genome sequence data analysed here also contain relatively little evidence for adaptive evolution, particularly on structural proteins, suggesting that most viral lineages are of similar fitness, and that WNV is well adapted to the ecology of mosquito vectors and diverse avian hosts in the USA. In sum, the molecular evolution of WNV in North America depicts a largely unfettered expansion within a permissive host and geographic population with little evidence of major adaptive barriers.

Journal of Virology

Ecological, morphological, genetic and life history characteristics of two sockeye salmon populations, Tustumena Lake, Alaska

Populations can differ in both phenotypic and molecular genetic traits. Phenotypic differences likely result from differential selection pressures in the environment, whereas differences in neutral molecular markers result from genetic drift associated with some degree of reproductive isolation. Two sockeye salmon, Oncorhynchus nerka, populations were compared using both phenotypic and genotypic characters, and causal factors were examined. Salmon spawning in a short (<3 km), shallow (<21 cm), clear, homogenous spring-fed study site spawned later, were younger, smaller, and produced fewer and smaller eggs than salmon spawning in a longer (∼80 km), deeper, stained, diverse, precipitation-dominated stream. Run timing differences were associated with differences in stream thermal regimes. Age and size at maturity differences are likely due to differences in age-specific mortality rates. Fish in the shallow spring-fed system suffered higher adult predation rates and exhibited greater egg to fry survival compared to fish in the precipitation-fed system. Salmon in both streams exhibited non-random nest site selection for deeper habitats and smaller substrates (≥2 to <64 mm mean diameter) relative to available habitat; fish from the precipitation system avoided low velocity habitats containing fine (<2 mm) substrates. Genetic comparisons of six microsatellite loci indicated that run time was a more effective reproductive isolating mechanism than geographical distance. Differences between and within the tributary spawning populations are discussed in terms of selection, genetic drift, and the homogenizing effects of gene flow. This study indicates important adaptive differences may exist between proximate spawning groups of salmon which should be considered when characterizing populations for conservation or management purposes.

Alaska

Ecological risk assessment of environmental stress and bioactive chemicals to riverine fish populations: An individual-based model of smallmouth bass Micropterus dolomieu

Ecological risk assessments play an important role in environmental management and decision-making. Although empirical measurements of the effects of habitat changes and chemical exposure are often made at molecular and individual levels, environmental decision-making often requires the quantification of management-relevant, population-level outcomes. In this study, we generalized a modeling framework to evaluate population-level ecological risk of environmental stress and bioactive chemicals. The modeling framework includes (1) a biological model module that incorporates complex and interacting biological and ecological processes, and environmental stochasticity, (2) an effect module that links the impacts of environmental changes and chemical exposure to individual characteristics, and (3) a population module that makes decisions on the choice of population-level properties to best capture the effects and thus to track in the model based on the target species and the research and management interest. This framework is a 3-module procedure that provides an alternative way for researchers to organize, present and communicate the risk assessment modeling studies. To demonstrate this framework, we used a socioeconomically important riverine fish species, smallmouth bass Micropterus dolomieu , as the model species. We developed an individual-based model as the biological model module. We evaluated the impacts of changing water temperature and flow regimes, and the impacts of exposure to estrogenic endocrine disrupting compounds (EEDC) on smallmouth bass populations in the Chesapeake Bay Watershed, USA. Warm summer water temperatures and year-round high flows had the most severe impacts on the smallmouth bass population. An increase in exposure level to EEDC, both year-round and in summer months, substantially reduced population size, spawner and recruit abundance, and the proportion of quality-length individuals. Acute exposure to EEDC was more detrimental to the population than chronic exposure. Acute exposure during spawning season had the most severe impacts. This modeling framework can be extended to other species, environmental factors and chemicals, and can be used to inform management and conservation decisions.

Delaware, Maryland, New York, Pennsylvania, Virgin

Puerto Rico Sicydium goby diversity: Species-specific insights on population structures and distributions

Sicydiine gobies are major contributors to Caribbean stream fish biodiversity, and ecosystem functions and services. In the Caribbean, Sicydiine gobies are represented by a single genus, Sicydium, but species-level Sicydium taxonomy and distributions remain unresolved in this region. A previous study posited that four species of Sicydium are present in Puerto Rico, including the recently described Sicydium gilberti; however, that study relied solely on morphological characters, and samples from a limited number of locations. Identification of Sicydium to species in studies with more comprehensive sampling has not occurred due to the impracticality of field identification. In our study, we employed morphological data and Mitochondrial Cytochrome C Oxidase I (mtCOI) genetic sequence data, collected from over 500 Sicydium spp. individuals at 16 broadly distributed sites, to provide an inventory of the species of Sicydium that occur in Puerto Rico. Additionally, we reveal new information on the spatial patterns of distribution, relative abundance, population structure, and observed molecular diversity among Sicydium taxa in Puerto Rico’s river systems. This information is essential to advance ecological and evolutionary knowledge of Caribbean fish assemblages, guide stream fish conservation, and for effective management of fisheries.

Neotropical Biodiversity

Accounting for rate variation among lineages in comparative demographic analyses

Genetic analyses of contemporary populations can be used to estimate the demographic histories of species within an ecological community. Comparison of these demographic histories can shed light on community responses to past climatic events. However, species experience different rates of molecular evolution, and this presents a major obstacle to comparative demographic analyses. We address this problem by using a Bayesian relaxed-clock method to estimate the relative evolutionary rates of 22 small mammal taxa distributed across northwestern North America. We found that estimates of the relative molecular substitution rate for each taxon were consistent across the range of sampling schemes that we compared. Using three different reference rates, we rescaled the relative rates so that they could be used to estimate absolute evolutionary timescales. Accounting for rate variation among taxa led to temporal shifts in our skyline-plot estimates of demographic history, highlighting both uniform and idiosyncratic evolutionary responses to directional climate trends for distinct ecological subsets of the small mammal community. Our approach can be used in evolutionary analyses of populations from multiple species, including comparative demographic studies.

Evolution

Experimental reintroduction of the federally endangered Santa Cruz Island bush mallow (Malacothamnus fasciculatus var. nestioticus)

Studies of Malacothamnus fasciculatus var. nesioticus were begun in 1995 to understand its distribution, reproductive biology and ecological requirements. After 100+ years of depredation by sheep, two known populations of fewer than 20 plants each survived in 1995. Molecular studies showed that each of the two populations was composed of 1–3 genets. During our study, two additional populations of similar size were discovered. Plants are self-compatible but require insect visitation to augment pollination. Based on seed set, viable embryos, and germination rates, we found no evidence for inbreeding depression. Bush mallow also reproduces vegetatively by rhizomes, the primary means of establishment and persistence in natural populations, and a key feature for maximizing recovery success. Ex situ observations and trial in situ outplantings suggested that supplemental watering was critical to initial survival. We developed a recovery strategy composed of four plots located at varying elevations and aspects. Each plot was enclosed to exclude feral pigs, which posed a continuing threat. Each plot was planted with twelve rooted plants derived from each of three natural populations. Plants were provided supplemental watering for four months. Survivorship after one year ranged from 46% to 91%. Significant differences in survivorship were observed relative to source of plants. After twelve months some plants were flowering and reproducing vegetatively.

California

Spatial and temporal variation in dissolved organic matter in urban streams in metropolitan Boston, Massachusetts (USA)

Urban riverine systems are heterogeneous, and the substantial variability in impervious cover, riparian cover, wetlands, and wastewater and stormwater infrastructure affect sources and transport of dissolved organic matter (DOM), of which dissolved organic C (DOC) is a substantial component. An understanding of the quantity, bioavailability, and timing of DOM inputs (a key energy source for food webs and a component of nutrient cycling) to streams within cities can help to better evaluate drivers of DOM variability. We sampled 100 stream sites in the greater Boston (Massachusetts, USA) area spanning a range of land cover, riparian vegetation, stream size, housing and infrastructure age, and sociodemographic characteristics. Water samples collected during 4 seasonal synoptic events in 2021 and 2022 were analyzed for DOC concentration and DOM characteristics (using fluorescence excitation–emission matrices and absorbance spectra). Temporally, we observed more-autochthonous DOM and lower DOC concentrations in the summer, possibly due to low precipitation and streamflow disconnecting streams from humic wetland and soil C sources. Consistent with other studies, we observed that more-urbanized streams had DOM that was less humic and more autotrophic. Higher wetland cover was associated with more-humic, higher-molecular-weight DOM and was the strongest predictor of DOM characteristics, suggesting that managers should consider the impacts of development on DOM, stream ecological functions, and CO 2 emissions. Interestingly, except during the very dry summer, sites downstream of combined sewer outfalls showed distinctly higher concentrations of protein-like DOM, suggesting the influence of sewage overflows and highlighting the potential for monitoring wastewater contamination using protein-like DOM. Although sociodemographic variables were not strong predictors of DOM composition, we observed a possible association between lower-income areas with less canopy cover and more-autochthonous DOM and between areas with older housing, more canopy cover, and more-humic DOM. These patterns suggest that equitable repair of wastewater infrastructure and restoration of riparian vegetation is needed.

Massachusetts

Influence of niche breadth and position on the historical biogeography of seafaring scincid lizards

Niche breadth and position can influence diversification among closely related species or populations, yet limited empirical data exist concerning the predictability of the outcomes. We explored the effects of these factors on the evolution of the Emoia atrocostata species group, an insular radiation of lizards in the western Pacific Ocean and Indo-Australasia composed of both endemic and widespread species that differ in niche occupancy. We used molecular data and phylogeographical diffusion models to estimate the timing and patterns of range expansion, and ancestral reconstruction methods to infer shifts in ecology. We show evidence of multidirectional spread from a centre of origin in western Micronesia, and that the phyletic diversity of the group is derived from a putative habitat specialist that survives in the littoral zone. This species is composed of paraphyletic lineages that represent stages or possible endpoints in the continuum toward speciation. Several descendant species have transitioned to either strand or interior forest habitat, but only on remote islands with depauperate terrestrial faunas. Our results suggest that the atrocostata group might be in the early phases of a Wilsonian taxon cycle and that the capacity to tolerate salt stress has promoted dispersal and colonization of remote oceanic islands. Divergence itself, however, is largely driven by geographical isolation rather than shifts in ecology.

Biological Journal of the Linnean Society