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Marine mammals are wide-ranging, relatively long-lived organisms that play a crucial role in maintaining healthy ocean ecosystems. Often referred to as ecosystem engineers and sentinel species in marine ecosystems, these charismatic megafauna feed at a variety of trophic levels, affecting food web dynamics and cycling of chemicals and nutrients in the water column as well as in benthic habitats, both nearshore and in the deep ocean. An understanding of their abundance and distribution is an essential starting point for evaluating their role in ocean ecosystems. Accordingly, marine mammals have been included among key variables to monitor in ocean observing systems, from core variablesfor the U.S. Integrated Ocean Observing System (IOOS) to an Essential Ocean Variable (EOV)for the Global Ocean Observing System (GOOS). They also contribute to several Essential Biodiversity Variables (EBVs)for the Group on Earth Observations Biodiversity Observation Network (GEO BON). Further, evaluation of the health of marine mammal populations will help deliver societal benefits by contributing to the UN Decade of Ocean Science for Sustainable Development; informing reporting activities such as the World Ocean Assessment; and supporting achievement of Sustainable Development Goal 14, the post-2020 framework for the Convention for Biological Diversity, and a new treaty for conservation and sustainable use of marine biodiversity beyond national jurisdiction. The goal of this Interagency Ocean Observation Committee task team was to advance the integration of existing biological observations from local, regional, and federal sources using best practices to inform national needs and ultimately feed seamlessly into the Global Ocean Observing System, as appropriate. To accomplish this goal the subgroup focused on marine mammals to: 1. Reconcile the IOOS core biological variables with GOOS EOVs and the GEO BON EBVs, identifying where there are clear synergies in terms of spatial and temporal observing requirements and existing observation infrastructure and delivery including best practices/standards. 2. Identify and improve pathways for data flow for observations of these variables from Federal sources, such as the stock assessments conducted by NMFS and FWS, into IOOS with a focus on identifying and implementing best practices surrounding standardized data collection and delivery adhering to the FAIR and CAREdata principles, as appropriate.

Correcting for amplification biases in genetic metabarcoding data can yield quantitative estimates of template DNA concentrations. However, a major source of uncertainty in metabarcoding data is the presence of non-detections, where a technical PCR replicate fails to detect a species observed in other replicates. Such non-detections are an important special case of variability among technical replicates in metabarcoding data, particularly in environmental samples. While many sampling and amplification processes underlie observed variation in metabarcoding data, understanding the causes of non-detections is an important step in distinguishing signal from noise in metabarcoding studies. Here, we use both simulated and empirical data to 1) develop a qualitative understanding of how non-detections arise in metabarcoding data, 2) outline steps to recognize uninformative data in practice, and 3) identify the conditions under which amplicon sequence data can reliably detect underlying biological signals. We show in both simulations and empirical data that, for a given species, the rate of non-detections among technical replicates is a function of both the template DNA concentration and species-specific amplification efficiency. Consequently, we conclude metabarcoding datasets are strongly affected by (1) deterministic amplification biases during PCR and (2) stochastic sampling of amplicons during sequencing — both of which we can model — but also by (3) stochastic sampling of rare molecules prior to PCR, which remains a frontier for quantitative metabarcoding. Our results highlight the importance of estimating species-specific amplification efficiencies and critically evaluating patterns of non-detection in metabarcoding datasets to better distinguish environmental signal from the noise inherent in molecular detections of rare targets.

Metabarcoding is a powerful molecular tool for simultaneously surveying hundreds to thousands of species from a single sample, underpinning microbiome and environmental DNA (eDNA) methods. Deriving quantitative estimates of underlying biological communities from metabarcoding is critical for enhancing the utility of such approaches for health and conservation. Recent work has demonstrated that correcting for amplification biases in genetic metabarcoding data can yield quantitative estimates of template DNA concentrations. However, a major source of uncertainty in metabarcoding data stems from non-detections across technical PCR replicates where one replicate fails to detect a species observed in other replicates. Such non-detections are a special case of variability among technical replicates in metabarcoding data. While many sampling and amplification processes underlie observed variation in metabarcoding data, understanding the causes of non-detections is an important step in distinguishing signal from noise in metabarcoding studies. Here, we use both simulated and empirical data to 1) suggest how non-detections may arise in metabarcoding data, 2) outline steps to recognize uninformative data in practice, and 3) identify the conditions under which amplicon sequence data can reliably detect underlying biological signals. We show with both simulations and empirical data that, for a given species, the rate of non-detections among technical replicates is a function of both the template DNA concentration and species-specific amplification efficiency. Consequently, we conclude metabarcoding datasets are strongly affected by (1) deterministic amplification biases during PCR and (2) stochastic sampling of amplicons during sequencing—both of which we can model—but also by (3) stochastic sampling of rare molecules prior to PCR, which remains a frontier for quantitative metabarcoding. Our results highlight the importance of estimating species-specific amplification efficiencies and critically evaluating patterns of non-detection in metabarcoding datasets to better distinguish environmental signal from the noise inherent in molecular detections of rare targets.

Marine heatwaves can drive large-scale shifts in marine ecosystems but studying their impacts on whole species assemblages can be difficult. Here, we leverage the taxonomic breadth and resolution of DNA sequences derived from environmental DNA (eDNA) in the ethanol of a set of 23-year longitudinal ichthyoplankton samples, combining these with microscopy-derived ichthyoplankton identification to yield higher-resolution, species-specific quantitative abundance estimates of fish assemblages in the California Current Large Marine Ecosystem during and after the 2014–16 Pacific marine heatwave. This integrated dataset reveals patterns of tropicalization with increases in southern, mesopelagic species and associated declines in important temperate fisheries targets (e.g., North Pacific Hake (Merluccius productus) and Pacific Sardine (Sardinops sagax)). We observed novel assemblages of southern, mesopelagic fishes and temperate species (e.g., Northern Anchovy, Engraulis mordax) even after the return to average water temperatures. Our innovative preservative derived eDNA metabarcoding and quantitative modeling approaches open the door to reconstructing the historical dynamics of assemblages from modern and archived samples worldwide.SummaryNovel quantitative abundance estimates from archived DNA reveals marine heatwave-associated shifts in fish assemblages.

Maximum lifespan of a species is the oldest that individuals can survive, reflecting the genetic limit of longevity in an ideal environment. Here we report methylation-based models that accurately predict maximum lifespan (r=0.89), gestational time (r=0.96), and age at sexual maturity (r=0.87), using cytosine methylation patterns collected from over 12,000 samples derived from 192 mammalian species. Our epigenetic maximum lifespan predictor corroborated the extended lifespan in growth hormone receptor knockout mice and rapamycin treated mice. Across dog breeds, epigenetic maximum lifespan correlates positively with breed lifespan but negatively with breed size. Lifespan-related cytosines are located in transcriptional regulatory regions, such as bivalent chromatin promoters and polycomb-repressed regions, which were hypomethylated in long-lived species. The epigenetic estimators of maximum lifespan and other life history traits will be useful for characterizing understudied species and for identifying interventions that extend lifespan.

SummaryEpigenetics has hitherto been studied and understood largely at the level of individual organisms. Here, we report a multi-faceted investigation of DNA methylation across 11,117 samples from 176 different species. We performed an unbiased clustering of individual cytosines into 55 modules and identified 31 modules related to primary traits including age, species lifespan, sex, adult species weight, tissue type and phylogenetic order. Analysis of the correlation between DNA methylation and species allowed us to construct phyloepigenetic trees for different tissues that parallel the phylogenetic tree. In addition, while some stable cytosines reflect phylogenetic signatures, others relate to age and lifespan, and in many cases responding to anti-aging interventions in mice such as caloric restriction and ablation of growth hormone receptors. Insights uncovered by this investigation have important implications for our understanding of the role of epigenetics in mammalian evolution, aging and lifespan.

Understanding diet is critical for conservation of endangered predators. Southern Resident killer whales (SRKW) (Orcinus orca) are an endangered population occurring primarily along the outer coast and inland waters of Washington and British Columbia. Insufficient prey has been identified as a factor limiting their recovery, so a clear understanding of their seasonal diet is a high conservation priority. Previous studies have shown that their summer diet in inland waters consists primarily of Chinook salmon (Oncorhynchus tshawytscha), despite that species’ rarity compared to some other salmonids. During other times of the year, when occurrence patterns include other portions of their range, their diet remains largely unknown. To address this data gap, we collected feces and prey remains from October to May 2004–2017 in both the Salish Sea and outer coast waters. Using visual and genetic species identification for prey remains and genetic approaches for fecal samples, we characterized the diet of the SRKWs in fall, winter, and spring. Chinook salmon were identified as an important prey item year-round, averaging ~50% of their diet in the fall, increasing to 70–80% in the mid-winter/early spring, and increasing to nearly 100% in the spring. Other salmon species and non-salmonid fishes, also made substantial dietary contributions. The relatively high species diversity in winter suggested a possible lack of Chinook salmon, probably due to seasonally lower densities, based on SRKW’s proclivity to selectively consume this species in other seasons. A wide diversity of Chinook salmon stocks were consumed, many of which are also at risk. Although outer coast Chinook samples included 14 stocks, four rivers systems accounted for over 90% of samples, predominantly the Columbia River. Increasing the abundance of Chinook salmon stocks that inhabit the whales’ winter range may be an effective conservation strategy for this population.

Harbor porpoise in the North Pacific are found in coastal waters from southern California to Japan, but population structure is poorly known outside of a few local areas. We used multiplexed amplicon sequencing of 292 loci and genotyped clusters of single nucleotide polymoirphisms as microhaplotypes (N = 271 samples) in addition to mitochondrial (mtDNA) sequence data (N = 413 samples) to examine the genetic structure from samples collected along the Pacific coast and inland waterways from California to southern British Columbia. We confirmed an overall pattern of strong isolation-by-distance, suggesting that individual dispersal is restricted. We also found evidence of regions where genetic differences are larger than expected based on geographical distance alone, implying current or historical barriers to gene flow. In particular, the southernmost population in California is genetically distinct (FST = 0.02 [microhaplotypes]; 0.31 [mtDNA]), with both reduced genetic variability and high frequency of an otherwise rare mtDNA haplotype. At the northern end of our study range, we found significant genetic differentiation of samples from the Strait of Georgia, previously identified as a potential biogeographical boundary or secondary contact zone between harbor porpoise populations. Association of microhaplotypes with remotely sensed environmental variables indicated potential local adaptation, especially at the southern end of the species' range. These results inform conservation and management for this nearshore species, illustrate the value of genomic methods for detecting patterns of genetic structure within a continuously distributed marine species, and highlight the power of microhaplotype genotyping for detecting genetic structure in harbor porpoises despite reliance on poor-quality samples.

Background Host-specific microbiomes play an important role in individual health and ecology; in marine mammals, epidermal microbiomes may be a protective barrier between the host and its aqueous environment. Understanding these epidermal-associated microbial communities, and their ecological- or health-driven variability, is the first step toward developing health indices for rapid assessment of individual or population health. In Cook Inlet, Alaska, an endangered population of beluga whales (Delphinapterus leucas) numbers fewer than 300 animals and continues to decline, despite more than a decade of conservation effort. Characterizing the epidermal microbiome of this species could provide insight into the ecology and health of this endangered population and allow the development of minimally invasive health indicators based on tissue samples. Results We sequenced the hypervariable IV region of bacterial and archaeal SSU rRNA genes from epidermal tissue samples collected from endangered Cook Inlet beluga whales (n = 33) and the nearest neighboring population in Bristol Bay (n = 39) between 2012 and 2018. We examined the sequences using amplicon sequence variant (ASV)-based analyses, and no ASVs were associated with all individuals, indicating a greater degree of epidermal microbiome variability among beluga whales than in previously studied cetacean species and suggesting the absence of a species-specific core microbiome. Epidermal microbiome composition differed significantly between populations and across sampling years. Comparing the microbiomes of Bristol Bay individuals of known health status revealed 11 ASVs associated with potential pathogens that differed in abundance between healthy individuals and those with skin lesions or dermatitis. Molting and non-molting individuals also differed significantly in microbial diversity and the abundance of potential pathogen-associated ASVs, indicating the importance of molting in maintaining skin health. Conclusions We provide novel insights into the dynamics of Alaskan beluga whale epidermal microbial communities. A core epidermal microbiome was not identified across all animals. We characterize microbial dynamics related to population, sampling year and health state including level of skin molting. The results of this study provide a basis for future work to understand the role of the skin microbiome in beluga whale health and to develop health indices for management of the endangered Cook Inlet beluga whales, and cetaceans more broadly.

The Southern Resident killer whale population (Orcinus orca) was listed as endangered in 2005 and shows little sign of recovery. Exposure to contaminants and risk of an oil spill are identified threats. Previous studies on contaminants have largely focused on legacy pollutants. Here we measure polycyclic aromatic hydrocarbons (PAHs) in whale fecal (scat) samples. PAHs are a diverse group of hazardous compounds (e.g., carcinogenic, mutagenic), and are a component of crude and refined oil as well as motor exhaust. The central finding from this study indicates low concentrations of the measured PAHs (

Taxonomy is an imprecise science that delimits the evolutionary continuum into discrete categories. For marine mammals, this science is complicated by the relative lack of morphological data for taxa that inhabit remote and often vast ranges. We provide guidelines to promote consistency in studies relying primarily on molecular genetic data to delimit cetacean subspecies from both populations and species. These guidelines identify informational needs: basis for the taxonomic hypothesis being tested, description of current taxonomy, description of relevant life history, sample distribution, sample size, number and sequence length of genetic markers, description of measures taken to ensure data quality, summary statistics for the genetic markers, and analytical methods used to evaluate the genetic data. We propose an initial set of quantitative and qualitative standards based on the types of data and analytical methods most readily available at present. These standards are not expected to be rigidly applied. Rather, they are meant to encourage taxonomic arguments that are consistent and transparent. We hope professional societies, such as the Society for Marine Mammalogy, will adopt quantitative standards that evolve as new data types and analytical methods become widely available.

Persistent organic pollutants (POPs), specifically PCBs, PBDEs, and DDTs, in the marine environment are well documented, however accumulation and mobilization patterns at the top of the food-web are poorly understood. This study broadens the understanding of POPs in the endangered Southern Resident killer whale population by addressing modulation by prey availability and reproductive status, along with endocrine disrupting effects. A total of 140 killer whale scat samples collected from 54 unique whales across a 4 year sampling period (2010-2013) were analyzed for concentrations of POPs. Toxicant measures were linked to pod, age, and birth order in genotyped individuals, prey abundance using open-source test fishery data, and pregnancy status based on hormone indices from the same sample. Toxicant concentrations were highest and had the greatest potential for toxicity when prey abundance was the lowest. In addition, these toxicants were likely from endogenous lipid stores. Bioaccumulation of POPs increased with age, with the exception of presumed nulliparous females. The exceptional pattern may be explained by females experiencing unobserved neonatal loss. Transfer of POPs through mobilization of endogenous lipid stores during lactation was highest for first-borns with diminished transfer to subsequent calves. Contrary to expectation, POP concentrations did not demonstrate an associated disruption of thyroid hormone, although this association may have been masked by impacts of prey abundance on thyroid hormone concentrations. The noninvasive method for measuring POP concentrations in killer whales through scat employed in this study may improve toxicant monitoring in the marine environment and promote conservation efforts.

Uncertainty in marine mammal taxonomy is increasingly being addressed using molecular genetic data. We examined 32 peer-reviewed articles published between 1994 and 2011to review methodological practices, consistency of markers and analytical methods, and overall quality of arguments used when genetic data have been employed to delimit new species and subspecies of marine mammals. The mitochondrial DNA (mtDNA) control region was the primary genetic marker used in these studies, but analytical methods varied greatly across studies. Diagnosability, a common metric for delimiting subspecies with morphological data, was only used through citing of fixed differences in mtDNA sequences. Assignment tests based on microsatellite data were less common but were applied at both taxonomic levels. Nuclear DNA sequence data were rarely used. Basic background material needed to evaluate the strength of arguments, such as distribution and sampling maps, were often missing. For most studies, sample sizes were good, but adequate geographic sampling for broadly distributed taxa was often lacking, diminishing the strength of evidence for taxonomic distinctness. Examining these empirical cases revealed a mixture of sound and inadequate practices for genetic studies of cetacean taxonomy and suggested that improvements could be made to the field by developing standard guidelines.

The Southern Resident killer whale population (Orcinus orca) was listed as endangered in 2005 and shows little sign of recovery. These fish eating whales feed primarily on endangered Chinook salmon. Population growth is constrained by low offspring production for the number of reproductive females in the population. Lack of prey, increased toxins and vessel disturbance have been listed as potential causes of the whale’s decline, but partitioning these pressures has been difficult. We validated and applied temporal measures of progesterone and testosterone metabolites to assess occurrence, stage and health of pregnancy from genotyped killer whale feces collected using detection dogs. Thyroid and glucocorticoid hormone metabolites were measured from these same samples to assess physiological stress. These methods enabled us to assess pregnancy occurrence and failure as well as how pregnancy success was temporally impacted by nutritional and other stressors, between 2008 and 2014. Up to 69% of all detectable pregnancies were unsuccessful; of these, up to 33% failed relatively late in gestation or immediately post-partum, when the cost is especially high. Low availability of Chinook salmon appears to be an important stressor among these fish-eating whales as well as a significant cause of late pregnancy failure, including unobserved perinatal loss. However, release of lipophilic toxicants during fat metabolism in the nutritionally deprived animals may also provide a contributor to these cumulative effects. Results point to the importance of promoting Chinook salmon recovery to enhance population growth of Southern Resident killer whales. The physiological measures used in this study can also be used to monitor the success of actions aimed at promoting adaptive management of this important apex predator to the Pacific Northwest.

Identifying demographic changes is important for understanding population dynamics. However, this requires long-term studies of definable populations of distinct individuals, which can be particularly challenging when studying mobile cetaceans in the marine environment. We collected photo-identification data from 19 years (1992-2010) to assess the dynamics of a population of bottlenose dolphins (Tursiops truncatus) restricted to the shallow (7 m) waters of Little Bahama Bank, northern Bahamas. This population was known to range beyond our study area, so we adopted a Bayesian mixture modeling approach to mark-recapture to identify clusters of individuals that used the area to different extents, and we specifically estimated trends in survival, recruitment, and abundance of a "resident" population with high probabilities of identification. There was a high probability (p= 0.97) of a long-term decrease in the size of this resident population from a maximum of 47 dolphins (95% highest posterior density intervals, HPDI = 29-61) in 1996 to a minimum of just 24 dolphins (95% HPDI = 14-37) in 2009, a decline of 49% (95% HPDI = approximately 5% to approximately 75%). This was driven by low per capita recruitment (average approximately 0.02) that could not compensate for relatively low apparent survival rates (average approximately 0.94). Notably, there was a significant increase in apparent mortality (approximately 5 apparent mortalities vs. approximately 2 on average) in 1999 when two intense hurricanes passed over the study area, with a high probability (p = 0.83) of a drop below the average survival probability (approximately 0.91 in 1999; approximately 0.94, on average). As such, our mark-recapture approach enabled us to make useful inference about local dynamics within an open population of bottlenose dolphins; this should be applicable to other studies challenged by sampling highly mobile individuals with heterogeneous space use.

For the majority of social species, group composition is dynamic, and individuals are interconnected in a heterogeneous social network. Social network structure has far-reaching implications for the ecology of individuals and populations. However, we have little understanding of how ecological variables shape this structure. We used a long-term data set (1984e2007) to examine the relationship between food availability and social network structure in the endangered southern resident killer whales. During the summer months individuals in this population feed primarily on chinook salmon, Oncorhynchus tshawytscha, which show annual variation in abundance. We tested the hypothesis that temporal variation in chinook salmon will correlate with variation in social network structure. Using a null model that controlled for population demography, group size and sampling effort, we found a significant relationship between the connectivity of the social network and salmon abundance, with a more interconnected social network in years of high salmon abundance. Our results demonstrate that resource availability may be an important determinant of social network structure. Given the central importance of the social network for population processes such as the maintenance of cooperation and the transmission of information and disease, a change in social network structure caused by a change in food availability may have significant ecological and evolutionary consequences. 2012 The Association for the Study of Animal Behaviour. Published by Elsevier Ltd. All rights reserved.

We used data from 78 individuals at 26 microsatellite loci to infer parental and sibling relationships within a community of fish-eating (‘‘resident’’) eastern North Pacific killer whales (Orcinus orca). Paternity analysis involving 15 mother/calf pairs and 8 potential fathers and whole-pedigree analysis of the entire sample produced consistent results. The variance in male reproductive success was greater than expected by chance and similar to that of other aquatic mammals. Although the number of confirmed paternities was small, reproductive success appeared to increase with male age and size. We found no evidence that males from outside this small population sired any of the sampled individuals. In contrast to previous results in a different population, many offspring were the result of matings within the same ‘‘pod’’ (long-term social group). Despite this pattern of breeding within social groups, we found no evidence of offspring produced by matings between close relatives, and the average internal relatedness of individuals was significantly less than expected if mating were random. The population’s estimated effective size was ,30 or about 1/3 of the current census size. Patterns of allele frequency variation were consistent with a population bottleneck.

Killer whales (Orcinus orca) currently comprise a single, cosmopolitan species with a diverse diet. However, studies over the last 30 yr have revealed populations of sympatric ‘‘ecotypes’’ with discrete prey preferences, morphology, and behaviors. Although these ecotypes avoid social interactions and are not known to interbreed, genetic studies to date have found extremely low levels of diversity in the mitochondrial control region, and few clear phylogeographic patterns worldwide. This low level of diversity is likely due to low mitochondrial mutation rates that are common to cetaceans. Using killer whales as a case study, we have developed a method to readily sequence, assemble, and analyze complete mitochondrial genomes from large numbers of samples to more accurately assess phylogeography and estimate divergence times. This represents an important tool for wildlife management, not only for killer whales but for many marine taxa. We used highthroughput sequencing to survey whole mitochondrial genome variation of 139 samples from the North Pacific, North Atlantic, and southern oceans. Phylogenetic analysis indicated that each of the known ecotypes represents a strongly supported clade with divergence times ranging from ;150,000 to 700,000 yr ago. We recommend that three named ecotypes be elevated to full species, and that the remaining types be recognized as subspecies pending additional data. Establishing appropriate taxonomic designations will greatly aid in understanding the ecological impacts and conservation needs of these important marine predators. Wepredict that phylogeographic mitogenomics will become an important tool for improved statistical phylogeography and more precise estimates of divergence times. [Supplemental material is available online at http://www.genome.org. The sequence data from this study have been submitted to GenBank (http://www.ncbi.nlm.nih.gov/genbank) under accession nos. GU187153–GU187164, GU187166– GU187219, and HM060332–HM060334.]

Quantitatively characterizing the social structure of a population provides important insight into the forces shaping key population processes. Moreover, long-term social dynamics provide an avenue for understanding population-level responses to changes in socioecological conditions. This is particularly true for species that show natal philopatry and highly stable hierarchically structured social units, such as the piscivorous resident killer whales of the northeast Pacific. The southern resident killer whale population is a small, demographically closed population, comprising three commonly recognized pods (J, K and L pods), that has recently been listed as endangered throughout its range in both Canadian and U.S.A. waters. In this study, we quantitatively assessed social structure in this population from 29 years of photo-identification data to characterize significant temporal changes in sociality. Preferential affiliation among killer whales within both genealogical matrilines and pods was supported by two different analytical methods and, despite interannual variability, these social clusters persisted throughout the study. All three pods experienced fluctuations in social cohesion over time, but the overall rate of intrapod affiliation was consistently lowest within L pod, the largest of the southern resident pods. The most recent increase in fluidity within social units, occurring in the mid to late 1990s, was coincident with a significant decline in population size, suggesting a possible common response to external stressors. Quantifying these trends in social structure is the first step towards understanding the causes and consequences of long-term changes in killer whale social structure.

We develop a Bayesian capture-recapture model that provides estimates of abundance as well as time-varying and heterogeneous survival and capture probability distributions. The model uses a state-space approach by incorporating an underlying population model and an observation model, and here is applied to photo-identification data to estimate trends in the abundance and survival of a population of bottlenose dolphins (Tursiops truncatus) in northeast Scotland. Novel features of the model include simultaneous estimation of time-varying survival and capture probability distributions, estimation of heterogeneity effects for survival and capture, use of separate data to inflate the number of identified animals to the total abundance, and integration of separate observations of the same animals from right and left side photographs. A Bayesian approach using Markov chain Monte Carlo methods allows for uncertainty in measurement and parameters, and simulations confirm the model's validity.

Data on morphometrics are central to understanding key features of both population and behavioral ecology, including phenotypic differences between species (Perryman and Lynn 1993, Meijaard and Groves 2004) and populations (Perryman and Lynn 1994), life history traits such as patterns of individual growth (Read et al. 1993, Lee and Moss 1995), and morphological differences related to mating systems (Boonstra et al. 1993, Tolley et al. 1995, Plavcan and van Schaik 1998). In the marine environment, it has proved difficult to obtain measurements from free-ranging cetaceans. Capture operations can yield precise data on morphometrics (e.g., Read et al. 1993), but such disruptive procedures are generally not feasible and are restricted to smaller species. On a more remote level, photogrammetric approaches have been successfully applied to estimate body size from aerial platforms (e.g., Perryman and Lynn 1993) and from underwater images (Klimley and Brown 1983, Spitz et al. 2000), but stereophotogrammetric methods require a camera configuration that is somewhat cumbersome (Klimley and Brown 1983, Brager et al. 1999, Brager and Chong 1999), and can be difficult to implement from small-boat research platforms alongside routine data collection. We describe a simple approach for obtaining morphometric measurements based on photographing two laser dots that have been projected onto the body of a whale using two small laser-pointers. These laser-pointers are mounted in a parallel orientation to maintain a fixed and known separation distance, and the dots therefore provide a scale of known dimension on the image of the whale that can be used to calibrate morphometric measurements. The lightweight laser setup can be conveniently mounted on a camera lens, allowing this approach to be implemented by a single photographer in conjunction with photo-identification studies. We demonstrate the

Population substructure has important implications for a species’ ecology and evolution. As such, knowledge of this structuring is critical for the conservation and management of natural populations. Among marine mammals, many examples exist of species that enjoy a broad geographical distribution, yet are characterized by fine-scale population subdivisions. Coastal bottlenose dolphins have been studied extensively in a few regions globally, and these studies have highlighted a great diversity in both social strategies and demographic isolation. Here we use molecular genetic markers to examine the degree of population subdivision among three study sites separated by less than 250 km on Little Bahama Bank in the northern Bahamas. Mitochondrial DNA (mtDNA) sequence variation and microsatellite

Assessment of pinniped predation most often relies on analysis of the hard, undigested prey remains evident in faecal (scat) samples. For many prey species this method can yield valuable information on predator–prey interactions. For some genera, however, species diagnostic characteristics are lost during the process of prey digestion, thereby preventing morphological identification of fish prey species. Here, the feasibility of using faecal DNA to detect the presence of salmonids in pinniped scat samples and to distinguish reliably between sea trout Salmo trutta and Atlantic salmon S. salar was assessed. Novel salmonid mitochondrial DNA (mtDNA) primers were designed to amplify 162 bp of the 16S rDNA and a 327 bp section of the cytochrome b gene. Species-specific banding patterns were obtained by digestion of the cytochrome b PCR product with the restriction endonuclease AluI, and confirmed by the species-specific amplification of the 16S rDNA fragment from Atlantic salmon. Scats collected from captive grey seals Halichoerus grypus fed on known monospecific diets used to validate the PCR-RFLP assay indicated a probability of at least 95.8% (23 of 24 faecal extracts) of detecting salmonids using DNA extracted from the scat matrix. Implemented alongside conventional prey remains analyses, this technique presents a promising new method for examining prey composition and assessing pinniped predation on salmonids.

Many studies have shown that the distribution of cetaceans can be closely linked to habitat, but the underlying function of the preferred habitats often remains unclear. Only when behavioural observations are made in relation to habitat types can functional mechanisms behind the habitat use be revealed. Within the range of a bottlenose dolphin (Tursiops truncatus) population off NE Scotland, dolphins show clear preferences for several discrete areas. If the observed patterns of distribution are related to foraging, we predict that behaviour patterns shown by dolphins would reflect this relationship. In this study we identify behaviours of dolphins at the water surface that were related to feeding events, evaluate whether the patterns of distribution were related to foraging and whether they were related the local submarine habitat characteristics. To investigate whether visible surface evidence of foraging behaviour varied spatially, we analysed data collected from 104 regular boat-based surveys made within the Moray Firth, NE Scotland, between 1990 and 2000. To determine whether underlying bathymetry had any influence on the surface behaviour of dolphins, a land-based observation study was carried out in the populations core region of use. The results of this study show that feeding behaviour by dolphins was significantly higher in areas used intensively by dolphins. Furthermore, there were clear relationships between feeding events and the submarine habitat characteristics; certain forms of feeding occur primarily over steep seabed gradients, and in deeper waters during June and July. These results quantitatively support the hypothesis that the distinctive patterns of distribution shown by these dolphins are related to foraging behaviour or opportunities, and that submarine habitat characteristics may be a significant factor in the foraging efficiency of dolphins. Future work should focus on collecting detailed information on the distribution patterns of prey within the study area to allow direct comparisons between predator and prey distributions.

Alliances between reproductive males have been described in both cetacean and terrestrial mammal societies, and kin selection theory has often been invoked to explain these stable male coalitions. However, recent studies of bottlenose dolphins, Tursiops aduncus, in Australia suggest that mechanisms other than kinship are fundamental in the formation of male alliances. We supplemented skin biopsy sampling with the collection of faecal samples from individually recognized wild bottlenose dolphins, T. truncatus, to assess the role of kin selection in alliance membership in the northeast Bahamas. Stable alliances between pairs of males were identified based on association analyses of individual photoidentification data collected over 4 years. Molecular genetic analyses of tissue samples revealed highly significant correlations between patterns of association and both mitochondrial DNA haplotype identity and microsatellite relatedness, indicating that males within long-term alliances are more closely related than expected by chance. These data reinforce the high degree of plasticity previously reported within the genus Tursiops, and suggest that social and ecological differences between populations can markedly affect the role of kin selection in determining social alliances between male bottlenose dolphins.  2003 Published by Elsevier Ltd on behalf of The Association for the Study of Animal Behaviour.

Biologic sample collection in wild cetacean populations is challenging. Most information on toxicant levels is obtained from blubber biopsy samples; however, sample collection is invasive and strictly regulated under permit, thus limiting sample numbers. Methods are needed to monitor toxicant levels that increase temporal and repeat sampling of individuals for population health and recovery models. The objective of this study was to optimize measuring trace levels (parts per billion) of persistent organic pollutants (POPs), namely polychlorinated-biphenyls (PCBs), polybrominated-diphenyl-ethers (PBDEs), dichlorodiphenyltrichloroethanes (DDTs), and hexachlorocyclobenzene, in killer whale scat (fecal) samples. Archival scat samples, initially collected, lyophilized, and extracted with 70 % ethanol for hormone analyses, were used to analyze POP concentrations. The residual pellet was extracted and analyzed using gas chromatography coupled with mass spectrometry. Method detection limits ranged from 11 to 125 ng/g dry weight. The described method is suitable for p,p'-DDE, PCBs-138, 153, 180, and 187, and PBDEs-47 and 100; other POPs were below the limit of detection. We applied this method to 126 scat samples collected from Southern Resident killer whales. Scat samples from 22 adult whales also had known POP concentrations in blubber and demonstrated significant correlations (p < 0.01) between matrices across target analytes. Overall, the scat toxicant measures matched previously reported patterns from blubber samples of decreased levels in reproductive-age females and a decreased p,p'-DDE/∑PCB ratio in J-pod. Measuring toxicants in scat samples provides an unprecedented opportunity to noninvasively evaluate contaminant levels in wild cetacean populations; these data have the prospect to provide meaningful information for vital management decisions.

Concern has been expressed over the status of the bottlenose dolphin population that uses the Moray Firth and adjacent waters in NE Scotland. Consequently, part of this population's range has been proposed as a Special Area of Conservation. Efforts to manage and monitor the status of this population require information on the level of genetic diversity within the population and its genetic relatedness to neighbouring populations to appropriately designate units for management and monitoring. Here we examine mitochondrial genetic diversity within the NE Scotland population, and compare this to other regions around the UK and Ireland. Sequence analysis of 549bp of the mitochondrial DNA control region identified eight unique haplotypes in a sample of 29 individuals. Analysis of molecular variance suggests that the Moray Firth population is genetically more closely related to Welsh animals than to its nearest neighbour population in west Scotland. Furthermore, measures of within-population genetic diversity were markedly lower in the Moray Firth than any other sampled region. The low levels of mtDNA genetic variability observed and its apparent geographic isolation provide further support for the precautionary approach currently being applied to the management of this population, despite the lack of direct evidence of harm.

S.D. Berrow *, B. Mchugh , D. Glynn , E. Mcgovern , K.M. Parsons , R.W. Baird , S.K. Hooker e a Shannon Dolphin and Wildlife Foundation, Merchants Quay, Co. Clare, Kilrush, Ireland b Marine Institute, Abbotstown, Castleknock, Dublin 15, Ireland c Department of Zoology, University of Aberdeen, Tillydrone Avenue, Aberdeen AB9 2TN, UK d Department of Biology, Dalhousie University, Halifax, NS, Canada B3M 4J1 e British Antarctic Survey, High Cross, Madingley Road, Cambridge CB3 0ET, UK

The research reported here was financially supported by the United States (U.S.) Navy Office of Naval Research (grant N000140710120; www.onr.navy.mil), the Naval Facilities Engineering Command (grants N002441110021, N002441210007 and contract N6660413P2671; www.navfac.navy.mil), the Strategic Environmental Research and Development Program (award RC-2114; www.serdp.org), and the NOAA Fisheries Acoustic Program (http://www.nmfs.noaa.gov/pr/acoustics/). Graduate student funding support for the first author was provided by the U.S. National Science Foundation Graduate Research Fellowship Program (www.nsfgrfp.org). The relevant data are available from Dryad at DOI: 10.5061/dryad.14v89. Dive capacity among toothed whales (suborder: Odontoceti) has been shown to generally increase with body mass in a relationship closely linked to the allometric scaling of metabolic rates. However, two odontocete species tagged in this study, the Blainville’s beaked whale Mesoplodon densirostris and the Cuvier’s beaked whale Ziphius cavirostris, confounded expectations of a simple allometric relationship, with exceptionally long (mean: 46.1 min & 65.4 min) and deep dives (mean: 1129 m & 1179 m), and comparatively small body masses (med.: 842.9 kg & 1556.7 kg). These two species also exhibited exceptionally long recovery periods between successive deep dives, or inter-deep-dive intervals (M. densirostris: med. 62 min; Z. cavirostris: med. 68 min). We examined competing hypotheses to explain observed patterns of vertical habitat use based on body mass, oxygen binding protein concentrations, and inter-deep-dive intervals in an assemblage of five sympatric toothed whales species in the Bahamas. Hypotheses were evaluated using dive data from satellite tags attached to the two beaked whales (M. densirostris, n = 12; Z. cavirostris, n = 7), as well as melon-headed whales Peponocephala electra (n = 13), short-finned pilot whales Globicephala macrorhynchus (n = 15), and sperm whales Physeter macrocephalus (n = 27). Body mass and myoglobin concentration together explained only 36% of the variance in maximum dive durations. The inclusion of inter-deep-dive intervals, substantially improved model fits (R2 = 0.92). This finding supported a hypothesis that beaked whales extend foraging dives by exceeding aerobic dive limits, with the extension of inter-deep-dive intervals corresponding to metabolism of accumulated lactic acid. This inference points to intriguing tradeoffs between body size, access to prey in different depth strata, and time allocation within dive cycles. These tradeoffs and resulting differences in habitat use have important implications for spatial distribution patterns, and relative vulnerabilities to anthropogenic impacts. Publisher PDF

Global climate change during the Late Pleistocene periodically encroached and then released habitat during the glacial cycles, causing range expansions and contractions in some species. These dynamics have played a major role in geographic radiations, diversification and speciation. We investigate these dynamics in the most widely distributed of marine mammals, the killer whale (Orcinus orca), using a global data set of over 450 samples. This marine top predator inhabits coastal and pelagic ecosystems ranging from the ice edge to the tropics, often exhibiting ecological, behavioural and morphological variation suggestive of local adaptation accompanied by reproductive isolation. Results suggest a rapid global radiation occurred over the last 350 000 years. Based on habitat models, we estimated there was only a 15% global contraction of core suitable habitat during the last glacial maximum, and the resources appeared to sustain a constant global effective female population size throughout the Late Pleistocene. Reconstruction of the ancestral phylogeography highlighted the high mobility of this species, identifying 22 strongly supported long-range dispersal events including interoceanic and interhemispheric movement. Despite this propensity for geographic dispersal, the increased sampling of this study uncovered very few potential examples of ancestral dispersal among ecotypes. Concordance of nuclear and mitochondrial data further confirms genetic cohesiveness, with little or no current gene flow among sympatric ecotypes. Taken as a whole, our data suggest that the glacial cycles influenced local populations in different ways, with no clear global pattern, but with secondary contact among lineages following long-range dispersal as a potential mechanism driving ecological diversification.

Noninvasive samples have proved useful in genotyping studies of free-ranging mammals. However, potential genotyping errors associated with such samples dictate the need for validation studies. This pilot study demonstrates the use of dolphin faeces in multilocus microsatellite genotyping studies. An empirical approach to calculating the rate of genotyping error was applied to data from matched pairs of blood or tissue and faecal samples from both captive and wild bottlenose dolphins. Microsatellite genotypes were assigned to dolphin faecal extracts with greater than 95% confidence by using a multiple tube approach, and at least two independent replicate genotypings.

The difficulties associated with detecting population boundaries have long constrained the conservation and management of highly mobile, wide-ranging marine species, such as killer whales (Orcinus orca). In this study, we use data from 26 nuclear microsatellite loci and mitochondrial DNA sequences (988 bp) to test a priori hypotheses about population subdivisions generated from a decade of killer whale surveys across the northern North Pacific. A total of 462 remote skin biopsies were collected from wild killer whales primarily between 2001 and 2010 from the northern Gulf of Alaska to the Sea of Okhotsk, representing both the piscivorous “resident” and the mammal-eating “transient” (or Bigg’s) killer whales. Divergence of the 2 ecotypes was supported by both mtDNA and microsatellites. Geographic patterns of genetic differentiation were supported by significant regions of genetic discontinuity, providing evidence of population structuring within both ecotypes and corroborating direct observations of restricted movements of individual whales. In the Aleutian Islands (Alaska), subpopulations, or groups with significantly different mtDNA and microsatellite allele frequencies, were largely delimited by major oceanographic boundaries for resident killer whales. Although Amchitka Pass represented a major subdivision for transient killer whales between the central and western Aleutian Islands, several smaller subpopulations were evident throughout the eastern Aleutians and Bering Sea. Support for seasonally sympatric transient subpopulations around Unimak Island suggests isolating mechanisms other than geographic distance within this highly mobile top predator.