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Guidelines for collecting and maintaining archives for genetic monitoring

Rapid advances in molecular genetic techniques and the statistical analysis of genetic data have revolutionized the way that populations of animals, plants and microorganisms can be monitored. Genetic monitoring is the practice of using molecular genetic markers to track changes in the abundance, diversity or distribution of populations, species or ecosystems over time, and to follow adaptive and non-adaptive genetic responses to changing external conditions. In recent years, genetic monitoring has become a valuable tool in conservation management of biological diversity and ecological analysis, helping to illuminate and define cryptic and poorly understood species and populations. Many of the detected biodiversity declines, changes in distribution and hybridization events have helped to drive changes in policy and management. Because a time series of samples is necessary to detect trends of change in genetic diversity and species composition, archiving is a critical component of genetic monitoring. Here we discuss the collection, development, maintenance, and use of archives for genetic monitoring. This includes an overview of the genetic markers that facilitate effective monitoring, describes how tissue and DNA can be stored, and provides guidelines for proper practice.

Conservation Genetics Resources

Characterization of the putatively introduced red alga Acrochaetium secundatum (Acrochaetiales, Rhodophyta) growing epizoically on the pelage of southern sea otters ( Enhydra lutris nereis )

Ecological associations between epibionts (organisms that live on the surface of another living organism) and vertebrates have been documented in both marine and terrestrial environments, and may be opportunistic, commensal, or symbiotic (Lewin et al. 1981, Holmes 1985, Allen et al. 1993, Bledsoe et al. 2006, Pfaller et al. 2008, Suutari et al. 2010). Although epibiont proliferation is frequently reported on slow-moving, sparsely haired organisms such as manatees and sloths, reports from densely furred, highly mobile mammals are much less common. There are reports of epizoic algae for several species of pinnipeds (Kenyon and Rice 1959, Scheffer 1962, Baldridge 1977, Allen et al. 1993), which rely to varying degrees on both pelage and blubber for thermoregulation, but the phenomenon has not been widely described. Scheffer (1962) noted that red algae was fairly common on the pelage of northern fur seals (Callorhinus ursinus), pinnipeds for which fur likely makes a comparatively high contribution to thermoregulation (Donohue et al. 2000). For species with pelage that plays a critical role of thermal insulation, it seems implausible that an epibiont would persist on healthy individuals that devote significant energy resources toward grooming and actively maintaining their coat. Biological characteristics of epibiont settlement and attachment, and physiological requirements of epizoic species play key roles in their successful colonization and potential host impacts. To investigate this relationship, we explore a novel discovery of an epizoic alga from southern sea otters, including describing algal development on sea otter hair and molecular identification of the algae.

Marine Mammal Science

Anticoagulant rodenticide toxicity to non-target wildlife under controlled exposure conditions

Much of our understanding of anticoagulant rodenticide toxicity to non-target wildlife has been derived from molecular through whole animal research and registration studies in domesticated birds and mammals, and to a lesser degree from trials with captive wildlife. Using these data, an adverse outcome pathway identifying molecular initiating and anchoring events (inhibition of vitamin K epoxide reductase, failure to activate clotting factors), and established and plausible linkages (coagulopathy, hemorrhage, anemia, reduced fitness) associated with toxicity, is presented. Controlled exposure studies have demonstrated that second-generation anticoagulant rodenticides (e.g., brodifacoum) are more toxic than first- and intermediate-generation compounds (e.g., warfarin, diphacinone), however the difference in potency is diminished when first- and intermediate-generation compounds are administered on multiple days. Differences in species sensitivity are inconsistent among compounds. Numerous studies have compared mortality rate of predators fed prey or tissue containing anticoagulant rodenticides. In secondary exposure studies in birds, brodifacoum appears to pose the greatest risk, with bromadiolone, difenacoum, flocoumafen and difethialone being less hazardous than brodifacoum, and warfarin, coumatetralyl, coumafuryl, chlorophacinone and diphacinone being even less hazardous. In contrast, substantial mortality was noted in secondary exposure studies in mammals ingesting prey or tissue diets containing either second- or intermediate-generation compounds. Sublethal responses (e.g., prolonged clotting time, reduced hematocrit and anemia) have been used to study the sequelae of anticoagulant intoxication, and to some degree in the establishment of toxicity thresholds or toxicity reference values. Surprisingly few studies have undertaken histopathological evaluations to identify cellular lesions and hemorrhage associated with anticoagulant rodenticide exposure in non-target wildlife. Ecological risk assessments of anticoagulant rodenticides would be improved with additional data on (i) interspecific differences in sensitivity, particularly for understudied taxa, (ii) sublethal effects unrelated to coagulopathy, (iii) responses to mixtures and sequential exposures, and (iv) the role of vitamin K status on toxicity, and significance of inclusion of supplemental vitamin K or menadione (provitamin) in the diet of test organisms. A more complete understanding of the toxicity of anticoagulant rodenticides in non-target wildlife would enable regulators and natural resource managers to better predict and even mitigate risk.

Book chapter

Combining individual and close-kin mark–recapture to design an effective wildlife population survey

Close-kin mark–recapture (CKMR) is a promising approach for assessing population size of species that have been difficult to survey using more traditional methods. Here, we combine individual and close-kin mark–recapture in a single modeling framework (ICKMR) and provide an example of study design using this approach for Pacific walrus ( Odobenus rosmarus divergens ). We develop the ICKMR model and test it using simulated datasets, then use properties of the pseudo-likelihood to investigate the expected precision in estimates of abundance with different proposed survey designs. Our motivating example, the Pacific walrus, is an ice-associated marine mammal found in the Bering and Chukchi seas, where it is an important resource for Indigenous peoples. Pacific walrus abundance declined in the late 20th century, and it is currently a species of conservation concern due to potential impacts of climate change, particularly the loss of sea ice. To reduce uncertainty in population size estimates, researchers undertook a genetic mark–recapture sampling campaign from 2013 to 2017 and collected tissue samples from over 8000 individuals. Another campaign of a similar scale is ongoing (2023–2028). While sample collection was designed for individual mark–recapture, advances in CKMR methods and associated molecular techniques mean that these samples could also be suitable for CKMR. The advantages of CKMR over mark–recapture include an increased effective sample size (because each individual tags itself and its parents, siblings, and offspring) and additional insights into demographic quantities of interest. To make best use of genetic samples, we combine individual mark–recapture (IMR) with CKMR (ICKMR) and investigate whether different sampling strategies can increase precision in estimates of abundance. Our modeling approach includes special considerations for walrus life history, including a multi-year inter-birth interval. We found that expected coefficients of variation (CVs) of the ICKMR estimates of abundance, adult female survival, juvenile female survival, and proportion of breeding females are lower than those expected from IMR alone, and with ICKMR, fewer years of sampling can be conducted to obtain sufficient precision in estimates of abundance. This work demonstrates the utility of ICKMR and could be applicable across a variety of taxa.

Ecology

Book review: Proceedings of the First International Snakehead Symposium

Snakehead fishes (family Channidae) are among the most maligned aquatic invasive species in the USA and some other countries where they have been introduced outside of their native range in Asia and Africa. Nevertheless, snakeheads continue to be widely exploited in the live‐food trade in aquaculture and wild‐capture fisheries, are highly sought by anglers, and are also popular in the aquarium trade (Courtenay and Williams 2004). The Northern Snakehead Channa argus is the most widespread of the three channid species that are currently naturalized in the USA. This species has generated much concern and controversy, a situation that is partly fueled by sensational media coverage and B‐grade science fiction horror films, such as “Frankenfish,” “Snakehead Terror,” and “Snakehead Swamp.” Media reports of snakehead introductions are often replete with provocative terms, such as “vicious,” “villain,” “voracious,” “monster,” “diabolical,” and even “ecological Armageddon.” When snakeheads first appeared in natural waters of the USA, fisheries professionals became increasingly interested in their status. Established populations rapidly expanded in the mid‐Atlantic region and Arkansas, with scattered reports of introduced snakeheads from isolated locations in Hawaii, California, North Carolina, Florida, the Upper Midwest, and New England. In 2002, snakeheads were added to the list of injurious fishes under the Lacey Act, thereby prohibiting their importation or transport across state lines without a permit. This symposium was conceived by the editors and other concerned fisheries professionals of the Mississippi River Basin Panel on Aquatic Invasive Species. The mission of the symposium, held in Alexandria, Virginia, in July 2018, was to bring together experts on snakehead biology and ecology and to synthesize existing information into summary papers. In this book, 35 authors contributed to 15 peer‐reviewed articles that detail the current state of knowledge about snakehead introductions in the USA. Additionally, 16 abstracts are included from meeting presentations that were not accompanied by full‐length manuscripts. Also included is a summary of a facilitated symposium panel discussion featuring eight experts representing state and federal natural resource agencies and private fishing organizations. The book is organized into six sections. In the first section (Distribution), three papers provide an overview of the Channa species introduced into the USA and historical accounts of occurrence and dispersal of the Northern Snakehead in the mid‐Atlantic region and Arkansas. The second section (Biology/Ecology) consists of two articles that examine growth and energetics of Northern Snakehead populations and two papers that investigate diet, diel feeding activity, and movement of this species in the Potomac River drainage. The third section (Monitoring/Response) includes a paper that models range expansion of the Northern Snakehead in the southeastern USA based on occurrence data and environmental conditions. Also included in this section is a paper summarizing an environmental DNA study to assess the status and range of the Bullseye Snakehead C. marulius in southern Florida. The fourth section (Management/Control) is comprised of four papers that address harvest, age and growth, and development of a stock–recruitment model to inform management decisions regarding control and mitigation for Northern Snakehead populations in the greater Chesapeake Bay area. The fifth section (Perspectives) includes a paper on the history of snakehead introductions in Japan and a thought‐provoking social commentary on the human dimensions of Northern Snakehead management. Abstracts in the final section provide brief summaries of a diversity of snakehead studies, including aspects of distribution, ecology, behavior, control and monitoring efforts, public outreach, and pathology. The summary of the panel discussion is an engaging dialogue about the challenges of snakehead management in the context of conflicts regarding snakeheads as injurious versus their value as game and food species. Most of this book is focused on the Northern Snakehead. Much has been done to document snakehead distributions and certain aspects of snakehead biology, such as diets, age, and growth. Less research has been devoted to understanding the ecological impacts of snakeheads to native aquatic communities and ecosystems. This book would have benefited from a chapter summarizing the current systematics and diversity of the Channidae to inform fisheries biologists about the morphological characteristics of the family, approximate numbers of genera and species, and taxonomic instability. Exemplifying the latter, recent molecular and morphological evidence indicates uncertainty regarding identification of the feral snakehead population in Florida (Adamson and Britz 2019). Those authors suggest that this population may have originated from western Thailand, a possibility that could have implications for understanding historical pathways of snakehead introductions into the USA. In comparison with many published AFS symposia, this volume is relatively narrow in scope and lacks cohesive integration. It will primarily be of interest to those fisheries professionals engaged in the study of snakeheads as well as other nonnative species for which there are contrasting social values regarding their management: whether to monitor and attempt control or eradication efforts or to maintain populations for harvest as game or food species. The book should serve to identify information gaps and guide future research.

Transactions of the American Fisheries Society

Gene expression and wildlife health: Varied interpretations based on perspective

We evaluated wildlife population health from the perspective of statistical means vs. variances. We outlined the choices necessary to provide the framework for our study. These consisted of spatial and temporal boundaries (e.g., choice of sentinel species, populations, time frame), measurement techniques (molecular to population level), and appropriate statistical analyses. We chose to assess the health of 19 sea otter populations, located in the north Pacific from the Aleutian Islands, AK, to Santa Barbara, CA, and varying in population growth rates and length of occupancy. Our focal metric was gene expression (i.e., mRNA transcripts) data that we had previously generated across sea otter populations as a measure of population health. We used statistical methods with different approaches (i.e., means vs. variances) and examined the subsequent interpretive outcomes and how these influence our assessment of “health.” Interpretations based on analyses using variances versus means overlapped to some degree. In general, sea otter populations with low variation in gene expression were limited by food resources and at or near carrying capacity. In populations where the variation in gene expression was moderate or high, four out of five populations were increasing in abundance, or had been recently increasing. Where we had additional information on sources of stressors at the level of the population, we were able to draw inferences from those stressors to specific gene expression results. For example, gene expression patterns of sea otters from Western Prince William Sound were consistent with long term exposure to petroleum hydrocarbons, whereas in Kachemak Bay, patterns were consistent with exposure to algal toxins. Ultimately, determination of population or ecosystem health will be most informative when multiple metrics are examined across disciplines in the context of specific scenarios and goals.

Alaska, British Columbia, California, Oregon, Wash