Otariid gammaherpesvirus 1 in South American fur seals (Arctocephalus australis) and a novel related herpesvirus in free-ranging South American sea lions (Otaria byronia): Prevalence and effects of age, sex, and sample type.

Otariid gammaherpesvirus 1 (OtGHV1) is associated with high rates of urogenital carcinoma in free-ranging California sea lions (Zalophus californianus; CSL), and until recently was reported only in the Northern Hemisphere. The objective of this study was to survey free-ranging South American sea lions (Otaria byronia; SASL) and South American fur seals (Arctocephalus australis: SAFS) in Punta San Juan, Peru for OtGHV1 and to determine prevalence characteristics. Twenty-one percent (14/67) of urogenital swabs collected over three years (2011, 2014, 2015) from live pinnipeds of both species tested positive with a pan-herpesvirus conventional PCR. Sequencing of SAFS amplicons revealed 100% homology to OtGHV1 at the DNA polymerase, glycoprotein B, and viral bcl2-like genes. Sequencing of SASL amplicons revealed a novel related virus, herein called Otariid gammaherpesvirus 8 (OtGHV8). For comparison of sample sites, urogenital, conjunctival, and oropharyngeal swabs collected from 136 live pinnipeds of both species at Punta San Juan between 2011-2018 were then assayed using quantitative PCR for a segment of the OtGHV1/8 DNA polymerase gene using a qPCR assay now determined to cross-react between the two viruses. In total, across both species, 38.6% (51/132) of urogenital swabs, 5.6% (4/71) of conjunctival swabs, and 1.1% (1/90) of oropharyngeal swabs were positive for OtGHV1/8, with SASL only positive on urogenital swabs. Results from SASL were complicated by the finding of OtGHV8, necessitating further study to determine prevalence of OtGHV1 versus OtGHV8 using an alternate assay. Results from SAFS suggest a potential relationship between OtGHV1 in SAFS and CSL. Though necropsy surveillance in SAFS is very limited, geographic patterns of OtGHV1-associated urogenital carcinoma in CSL and the tendency of herpesviruses to cause more detrimental disease in aberrant hosts suggests that it is possible that SAFS may be the definitive host of OtGHV1, which gives further insight into the diversity and phyogeography of this clade of related gammaherpesviruses.

MEASUREMENT OF PERSISTENT ORGANIC POLLUTANTS, PERFLUORINATED COMPOUNDS, AND TOXIC METALS IN THE BLOOD OF HUMBOLDT PENGUINS (SPHENISCUS HUMBOLDTI) AT PUNTA SAN JUAN, PERU USING DRIED BLOOD SPOTS.

The Humboldt penguin (Spheniscus humboldti) population at the Punta San Juan Marine Protected Area in Peru is considered critical to the long-term sustainability of this endangered species in Peru. Exposure of the rookery to environmental toxicants is a mounting concern because of regional growth of industries and human populations. Whole blood samples were collected from 30 free-ranging penguins in 2011 as part of a broader population health monitoring program. Dried blood spots (DBS) containing 50 µl of blood were prepared and analyzed to assess exposure to five groups of environmental contaminants. Concentrations of elements arsenic, cadmium, iron, lead, mercury, selenium, and thallium were analyzed using inductively coupled plasma mass spectrometry. Persistent organic pollutant concentrations were measured using gas chromatography-tandem mass spectrometry to analyze organochlorine pesticides (OCP; p,p'-DDT, p,p'-DDE, ß-hexachlorocyclohexane, t-nonachlor, and oxychlordane), polychlorinated biphenyls (congeners 138 and 153), and polybrominated flame retardants (polybrominated biphenyl-153 and polybrominated diphenyl ether congeners 47 and 99). Per- and polyfluoroalkyl substances, including perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid were measured using liquid chromatography-tandem mass spectrometry. Results revealed low levels of exposure to these selected contaminants, at levels not considered to be of concern for wildlife health. DBS methodology was considered effective in a field-based setting for quantification of whole blood concentrations of environmental contaminants in penguins.

Genomic evidence for homoploid hybrid speciation in a marine mammal apex predator.

Hybridization is widespread and constitutes an important source of genetic variability and evolution. In animals, its role in generating novel and independent lineages (hybrid speciation) has been strongly debated, with only a few cases supported by genomic data. The South American fur seal (SAfs) Arctocephalus australis is a marine apex predator of Pacific and Atlantic waters, with a disjunct set of populations in Peru and Northern Chile [Peruvian fur seal (Pfs)] with controversial taxonomic status. We demonstrate, using complete genome and reduced representation sequencing, that the Pfs is a genetically distinct species with an admixed genome that originated from hybridization between the SAfs and the Galapagos fur seal (Arctocephalus galapagoensis) ~400,000 years ago. Our results strongly support the origin of Pfs by homoploid hybrid speciation over alternative introgression scenarios. This study highlights the role of hybridization in promoting species-level biodiversity in large vertebrates.

Agreement Study between Total Leukocyte Count Methodologies in South American Sea Lions (Otaria byronia) and Peruvian Fur Seals (Arctocephalus australis).

South American sea lions (Otaria byronia) and Peruvian fur seals (Arctocephalus australis) are sympatric species inhabiting the coastal Peruvian marine ecosystem. Declining abundance has prompted population health monitoring programs, including temporal monitoring of blood parameters. Several methods exist to determine total leukocyte count, yet no studies have evaluated agreement between methods in pinnipeds. We assessed agreement between total leukocyte counts determined by blood film estimate, Leuko-TIC, HemoCue, and UNOPETTE methods by using archival results from pinnipeds at Punta San Juan, Peru. Blood film estimates were prospectively performed, and resulting data were compared with retrospective leukocyte counts obtained from both species between 2009 and 2019 by using the other methodologies. Agreement in hematologic counts between methods was evaluated using Passing-Bablok regression and Bland-Altman plots (α=0.05). In total, 295 individuals (201 A. australis and 94 O. byronia) were included in the analysis. The blood film estimate method resulted in the highest leukocyte values (P<0.0001). Leuko-TIC counts were significantly higher than HemoCue counts (P<0.0001). Constant and proportional error was present in the agreement between the blood film estimate method and the other methods. Given the variation demonstrated between the different methodologies, additional research is needed to further evaluate agreement between these methodologies. The results underscore the importance of maintaining consistency in leukocyte count methodology for monitoring trends in population health over time. Method consistency may be the more important clinical consideration for assessing changes in leucocyte count over time and avoiding apparent changes depending on the methodology used.

DETECTION AND PREVALENCE OF SPHENISCID ALPHA-HERPESVIRUS-1 (SpAHV-1) IN A SAMPLE OF HUMBOLDT PENGUINS (SPHENISCUS HUMBOLDTI) AT PUNTA SAN JUAN, PERU.

Infections with herpesvirus have contributed to respiratory, enteric, and neurological disease reports in avian species worldwide. Herpesviruses have been detected in penguin species before but have not been studied extensively. To better understand the impact of these viruses in free-living populations, an initial retrospective survey was performed on a wild population of Humboldt penguins (Spheniscus humboldti) in the Punta San Juan Marine Protected Area, Peru (15°22'S, 75°12'W) using tracheal swabs collected from 28 penguins in 2016 and 34 penguins in 2018. DNA extracted from these swabs was analyzed using a consensus herpesviral PCR assay targeting the DNA polymerase gene, and positive samples were sequenced. A single sample from 2016 was positive for spheniscid alpha-herpesvirus-1 (SpAHV-1), establishing an overall sample prevalence of 1.6% (95% CI: 0-8.6%). The positive animal was an adult male that did not show any clinical signs of herpesviral infection and was otherwise healthy based on physical exam and laboratory findings. This is the first detection of a herpesvirus in penguins at Punta San Juan, Peru, and the first step toward characterizing the implications of SpAHV-1 for Humboldt penguins. This investigation highlights the importance of continual disease surveillance in wild populations over time to monitor for changes that may impact long-term population viability.

Terrestrial mammals of the Americas and their interactions with plastic waste.

Plastics have brought many benefits to society, but their mismanagement has turned them into a serious environmental problem. Today, the effects of plastic waste on wildlife are becoming increasingly evident. Since studies on plastic pollution have focused on species in marine ecosystems, here we review current knowledge on interactions between terrestrial mammals and plastic waste in the countries of the Americas, which is a global hotspot of mammalian biodiversity and in turn has, among its member countries, nations with high per capita generations of plastic waste globally. We identified 46 scientific articles documenting plastic ingestion in 37 species and four species that used plastic waste for nest or burrow construction. Of the 46 investigations, seven focused on plastic contamination, while the others reported on the presence of plastics in wildlife, even though this was not the primary focus of the research. However, these publications lack analytical methods commonly used in plastic studies, and only one study applied a standardized methodology for plastic detection. Therefore, in general, plastic pollution research on terrestrial mammals is limited. We extend several recommendations such as designing methodologies that are adapted to terrestrial mammals for the identification of plastics in fecal matter or gastrointestinal contents, carrying out species-specific analyzes on the impacts of plastics in nests or burrows, and giving further attention to this understudied issue and taxa.

Sympatric otariids increase trophic segregation in response to warming ocean conditions in Peruvian Humboldt Current System.

Determining trophic habits of predator communities is essential to measure interspecific interactions and response to environmental fluctuations. South American fur seals, Arctocephalus australis (SAFS) and sea lions Otaria byronia (SASL), coexist along the coasts of Peru. Recently, ocean warming events (2014-2017) that can decrease and impoverish prey biomass have occurred in the Peruvian Humboldt Current System. In this context, our aim was to assess the effect of warming events on long-term inter- and intra-specific niche segregation. We collected whisker from SAFS (55 females and 21 males) and SASL (14 females and 22 males) in Punta San Juan, Peru. We used δ13C and δ15N values serially archived in otariid whiskers to construct a monthly time series for 2005-2019. From the same period we used sea level anomaly records to determine shifts in the predominant oceanographic conditions using a change point analysis. Ellipse areas (SIBER) estimated niche width of species-sex groups and their overlap. We detected a shift in the environmental conditions marking two distinct periods (P1: January 2005-October 2013; P2: November 2013-December 2019). Reduction in δ15N in all groups during P2 suggests impoverished baseline values with bottom-up effects, a shift towards consuming lower trophic level prey, or both. Reduced overlap between all groups in P2 lends support of a more redundant assemblage during the colder P1 to a more trophically segregated assemblage during warmer P2. SASL females show the largest variation in response to the warming scenario (P2), reducing both ellipse area and δ15N mean values. Plasticity to adapt to changing environments and feeding on a more available food source without fishing pressure can be more advantageous for female SASL, albeit temporary trophic bottom-up effects. This helps explain larger population size of SASL in Peru, in contrast to the smaller and declining SAFS population.

Impact of marine debris recorded in a sympatric colony of otariids in the south coast of Peru.

Marine debris, in particular plastic debris and debris related to fishing activities, have an impact on marine mammals and seabirds. In marine-coastal species such as otariids, we can detect the incidence of these interactions through land-based observations. In this study, we recorded and described interactions between marine debris and the otariids Arctocephalus australis and Otaria byronia in a natural protected area in Peru, according to age class categories. Between 2016 and 2020, we recorded 151 animals that were directly impacted and/or were observed interacting with marine debris. Residual collars from fishing gear, ropes, plastic bags and unidentified material were recorded. Direct interactions with debris at colonies (e.g., plastic bags) were also recorded, as well as animals evidenced interacting with hooks. The age class category and species with the highest number of individuals impacted by residual collars were adult female A. australis and subadult male O. byronia. Single-use plastics were the most commonly bitten/moved waste by animals in the reserve. In the case of hooks, these were mostly lodged in the mouths of otariids. Our results highlight the importance of continuing to promote segregation at source, as well as the correct disposal of waste, locally and globally, to mitigate impacts on marine fauna.

Adenovirus Diversity in Fur Seal and Penguin Colonies of South America.

Adenoviruses are medium size nonenveloped viruses with a trend of coevolution with their hosts. We surveyed South American fur seals (Arctocephalus australis) and Humboldt penguins (Spheniscus humboldti) for adenoviruses at two sites from 2009 to 2012. Despite the common pattern of host specificity, some of the adenoviruses in our study were present in samples from unexpected host species. We identified mastadenoviruses, aviadenoviruses, and siadenoviruses in A. australis from Peru and Chile and in S. humboldti from Peru. The El Niño Southern Oscillation (ENSO) significantly reduces the productivity of the Humboldt upwelling system, which can change trophic and other ecological interactions, facilitating exposure to new pathogens. One aviadenovirus was detected in both the penguins and the fur seals, an interclass distance. This finding occurred only during the 2009 ENSO and not in 2010 or 2012. Further studies of viral diversity in sites with high-density mixed species populations are necessary to better understand viral evolution and the effect of environmental change on viral evolution and host specificity.

Trends in sympatric otariid populations suggest resource limitations in the Peruvian Humboldt Current System.

Sympatric species evolve mechanisms to avoid competition and coexist. In the Humboldt Current System (HCS), populations of South American sea lions (SASL, Otaria byronia) and South American fur seals (SAFS, Arctocephalus australis) fluctuate mostly due to ENSO events and prey availability. We evaluate population trajectories of Peruvian sympatric otariids and discuss mechanisms for competition and/or resource limitation. For this purpose, we analyzed population trajectories of SASL and SAFS in a sympatric breeding site in Punta San Juan, Peru between 2001 and 2019. Wavelet analysis was used to extract trends and derivatives to estimate rates and turning points. Age-class proportions and biomass times series were constructed from weekly counts and evaluated. Both populations show a growth phase and subsequent decline. SAFS started to decline ~2.25 years before and at a rate 1.5 times faster than SASL. Decrease in juvenile age-class suggests that resource limitation is the main contributing factor for current population decline.

Phylogenomic Discordance in the Eared Seals is best explained by Incomplete Lineage Sorting following Explosive Radiation in the Southern Hemisphere.

The phylogeny and systematics of fur seals and sea lions (Otariidae) have long been studied with diverse data types, including an increasing amount of molecular data. However, only a few phylogenetic relationships have reached acceptance because of strong gene-tree species tree discordance. Divergence times estimates in the group also vary largely between studies. These uncertainties impeded the understanding of the biogeographical history of the group, such as when and how trans-equatorial dispersal and subsequent speciation events occurred. Here, we used high-coverage genome-wide sequencing for 14 of the 15 species of Otariidae to elucidate the phylogeny of the family and its bearing on the taxonomy and biogeographical history. Despite extreme topological discordance among gene trees, we found a fully supported species tree that agrees with the few well-accepted relationships and establishes monophyly of the genus Arctocephalus. Our data support a relatively recent trans-hemispheric dispersal at the base of a southern clade, which rapidly diversified into six major lineages between 3 and 2.5 Ma. Otaria diverged first, followed by Phocarctos and then four major lineages within Arctocephalus. However, we found Zalophus to be nonmonophyletic, with California (Zalophus californianus) and Steller sea lions (Eumetopias jubatus) grouping closer than the Galapagos sea lion (Zalophus wollebaeki) with evidence for introgression between the two genera. Overall, the high degree of genealogical discordance was best explained by incomplete lineage sorting resulting from quasi-simultaneous speciation within the southern clade with introgresssion playing a subordinate role in explaining the incongruence among and within prior phylogenetic studies of the family. [Hybridization; ILS; phylogenomics; Pleistocene; Pliocene; monophyly.].

MEASUREMENT OF INTRAOCULAR PRESSURE USING REBOUND TONOMETRY IN ANESTHETIZED PERUVIAN FUR SEALS (ARCTOCEPHALUS AUSTRALIS) FROM PUNTA SAN JUAN, PERU.

Ocular disease in pinnipeds under human care is well described, and intraocular pressure (IOP) can be impacted by a variety of ophthalmic conditions. Species-specific reference parameters from clinically normal animals are instrumental for understanding how ophthalmic diseases may impact ocular pressures. IOP measurements were obtained using rebound tonometry from free-ranging Peruvian fur seals (Arctocephalus australis unnamed subspecies) at Punta San Juan, Peru, over a 6-yr period (2010-2016). Retrospective data obtained from 108 (81 adults and 27 neonates comprising 69 females and 39 males) anesthetized fur seals with normal anterior segment ophthalmic examinations was included in the analysis. Differences in IOP from each eye were compared to categorical variables (age, year, sex, restraint) using an independent-samples t test. All univariate results with a significance of P < 0.05 were included in multivariate analysis. Of the 13 general linear models evaluated, the top two for both the right and the left eye included age class when all variables were evaluated simultaneously. Neonates had significantly lower IOP values than adults in both the right eye (17.5 mm Hg; 95% confidence interval [CI]: 14.0-21.1 mm Hg compared to 33.5 mm Hg; 95% CI: 31.0-36.1 mm Hg, respectively) and the left eye (18.4 mm Hg; 95% CI: 14.4-22.5 mm Hg compared to 32.3 mm Hg; 95% CI: 29.3-35.3 mm Hg, respectively). Anesthesia method was not statistically significant (P > 0.05). This is the first report of normal IOP measurements for any fur seal species. Described data can be used to improve diagnosis and management of ocular alterations in pinnipeds.

Circulating nutrient concentrations in free-ranging Humboldt penguins (Spheniscus humboldti) in Punta San Juan, Peru.

Free-ranging Humboldt penguin (HP, Spheniscus humboldti) populations are under pressure from resource competition with industrial fisheries, habitat loss, and El Niño Southern Oscillation events. Foraging patterns for this top marine predator change during periods of aberrant oceanographic conditions and scarce fish stock numbers. These radical dietary fluctuations can lead to poor fertility, early embryonic death, poor hatchability, suppressed immune function, high chick mortality, and illness. To understand the variability of nutrient status in reproductive seasons, we measured select circulating nutrient concentrations (fat-soluble vitamins A, D, E, and carotenoids, fatty acids, amino acids, minerals, and electrolytes) of 105 HP at Punta San Juan, Peru during the first reproductive seasons of 2007 and 2008. We determined significant differences in nutrient status between sexes, years of sampling, and reproductive stages. Males (4.5 ± 0.38 kg) weighed more than females (4.0 ± 0.29 kg) and exhibited higher concentrations of vitamin A (0.71 ± 0.11 vs. 0.61 ± 0.12 µg/ml) and docosahexaenoic acid (6.70 ± 1.61 vs. 5.65 ± 1.59%). Males also displayed lower concentrations of ß-carotene (0.01 ± 0.01 vs. 0.012 ± 0.001 µg/ml) and phosphorus (3.43 ± 0.83 vs. 4.40 ± 1.66 mg/dl). Comparison between the 2 years showed most circulating amino acid concentrations were higher in 2007. Significant differences in circulating amino acids and vitamins were also noted between different reproductive stages. These results demonstrate concentrations of nutrients can vary due to the physiological state of the animal, as well as the overall dynamics of their marine ecosystem habitat.

Monitoring the occurrence of microplastic ingestion in Otariids along the Peruvian and Chilean coasts.

Repeated reports of microplastic pollution in the marine pinniped diet have emerged in the last years. However, only few studies address the drivers of microplastics presence and the potential implications for monitoring microplastic pollution in the ocean. This study monitored their in the scats (N = 205) of four pinniped species/subspecies at five different locations in the southern Pacific Ocean (Peru and Chile). Samples from all rookeries contained microplastics, and overall, 68% of the examined scats contained fragments/fibers, mostly blue colored. We confirmed that 81.5% of the fragments/fibers were anthropogenic in origin , but only 30% were polymers. Scats from Juan Fernández Archipelago presented higher microplastic concentrations than continental rookeries. Also, the common diet in each location may influence the levels found in the samples. This study presents a useful non-invasive technique to track plastic pollution in top predator diets as bioindicators for future surveillance/management plans applied to different location.

Inter-annual isotopic niche segregation of wild humboldt penguins through years of different El Niño intensities.

The Humboldt Current System presents high interannual variability, influenced by the El Niño Southern Oscillation (ENSO), whose implications in wildlife are not fully understood. We studied the isotopic niche of wild Humboldt penguins at Punta San Juan (Peru) during the pre-moult foraging trip in 4 consecutive years (2008-2011) under known oceanographic (ENSO) conditions. Our results show that there is a clear isotopic niche segregation (on both δ13C and δ15N values) of wild Humboldt penguins among all years. Besides isotopic niche segregation, niche width also varied significantly among years. The larger isotopic niche displayed in 2008 reflected the opportunistic feeding behaviour of Humboldt penguins when oceanographic conditions were unfavourable (i.e. El Niño of strong intensity). In contrast, and despite strong segregation, penguins displayed a more specialist behaviour in years of mild environmental conditions (i.e. 2009 "warm-weak", 2010 "neutral" and 2011 "warm-moderate"). No evidence of sexual segregation in wild Humboldt penguins during the pre-moulting foraging trip was found. This study highlights the coping mechanisms of an endangered species to changes in environmental conditions (i.e. overall, from strong to neutral El Niño events), which should have important ramifications in the management of the marine ecosystem in Peru, particularly the one related to the anchovy industry.

Measurement of Intraocular Pressure Using Rebound Tonometry in Anesthetized Free-ranging South American Sea Lions (Otaria byronia).

Intraocular pressures (IOPs) were measured using a rebound tonometer in 56 free-ranging adult South American sea lions (Otaria byronia) from Punta San Juan, Peru. All animals were anesthetized using medetomidine, midazolam, and butorphanol and determined to be in good health. No ocular abnormalities were observed affecting the cornea or ocular adnexa. Field conditions precluded evaluation of the lens and posterior segment of the eye. Mean (SD) IOP values for males (n=37) were 31±11 mmHg (right eye) and 31±9 mmHg (left eye). The same values for females (n=19) were 24±9 mmHg in the left eye and 27±10 mmHg in the right eye. These values are similar to those previously reported for pinnipeds under professional care and higher than those generally reported for terrestrial mammals.

Analysis of Toxic and Essential Elements in the Blood and Feathers of Humboldt Penguins ( Spheniscus humboldti) at Punta San Juan, Peru.

Whole blood, serum, and feather samples from 29 Humboldt Penguins ( Spheniscus humboldti) at the Punta San Juan Marine Protected Area, Peru, were analyzed for 55 toxic and essential elements by using inductively coupled plasma mass spectrometry. Mercury (Hg) was analyzed by cold vapor atomic fluorescence. Maximum Hg concentrations in serum (0.0056 mg/g), whole blood (0.297 mg/kg), and feathers (1.8 mg/kg dry weight) were at levels generally not considered to cause health impairment. Of the elements analyzed, only eight (aluminum, calcium, iron, Hg, potassium, magnesium, sodium, and zinc) were detected in serum. These elements, plus selenium and titanium, were also quantifiable in whole blood. Feather analysis detected quantifiable values for the elements found in serum, plus arsenic, boron, barium, copper, manganese, and titanium. Results indicate this important breeding population of endangered penguins did not appear to be exposed to environmental elemental contaminants at levels detrimental to health and reproductive success. However, identification of measurable concentrations of toxic elements at low levels underscores the need for continued environmental monitoring, particularly in the face of expanding regional human populations and industrial growth. These results provide important reference data for temporospatial monitoring of this and other penguin populations.

Helminth and respiratory mite lesions in Pinnipeds from Punta San Juan, Peru.

The tissues and parasites collected from Peruvian fur seals (Arctocephalus australis) and South American sea lions (Otaria byronia) found dead at Punta San Juan, Peru were examined. The respiratory mite, Orthohalarachne attenuata infected 3 out of 32 examined fur seals and 3 out of 8 examined sea lions, however caused moderate to severe lymphohistiocytic pharyngitis only in fur seals. Hookworms, Uncinaria sp, infected 6 of the 32 examined fur seals causing variable degrees of hemorrhagic and eosinophilic enteritis. This parasite caused the death of 2 of these pups. In fur seals and sea lions, Corynosoma australe and Contracaecum osculatum were not associated with significant tissue alterations in the intestine and stomach respectively. Respiratory mites and hookworms have the potential to cause disease and mortality among fur seals, while parasitic infections do not impact significatively the health of sea lions at Punta San Juan, Peru.

ASSESSMENT OF POLYCHLORINATED BIPHENYLS, ORGANOCHLORINE PESTICIDES, AND POLYBROMINATED DIPHENYL ETHERS IN THE BLOOD OF HUMBOLDT PENGUINS ( SPHENISCUS HUMBOLDTI) FROM THE PUNTA SAN JUAN MARINE PROTECTED AREA, PERU.

Persistent organic pollutants were assessed in Humboldt Penguins ( Spheniscus humboldti) from the Punta San Juan Marine Protected Area, Peru, in the austral winter of 2009. Plasma samples from 29 penguins were evaluated for 31 polychlorinated biphenyl (PCB) congeners and 11 organochlorine pesticides (OCPs) by using gas chromatography coupled to an ion trap mass spectrometer and for 15 polybrominated diphenyl ether (PBDE) congeners by using gas chromatography coupled with high-resolution mass spectrometry. The detection rate for PCBs in the samples was 69%, with congeners 105, 118, 180, and 153 most commonly detected. The maximum ΣPCB concentration was 25 ng/g. The detection rate for DDT, DDD, and/or DDE was higher than for other OCP residues (90%; maximum concentration=10 ng/g). The detection rate for PBDEs was 86%, but most concentrations were low (maximum ΣPBDE concentration=3.81 ng/g). This crucial breeding population of S. humboldti was not exposed to contaminants at levels detrimental to health and reproductive success; however, the identified concentrations of legacy and recently emerged toxicants underscore the need for temporal monitoring and diligence to protect this endangered species in the face of regional human population and industrial growth. These results also provide key reference values for spatial comparisons throughout the range of this species.

Ancient female philopatry, asymmetric male gene flow, and synchronous population expansion support the influence of climatic oscillations on the evolution of South American sea lion (Otaria flavescens).

The South American sea lion (Otaria flavescens) is widely distributed along the southern Atlantic and Pacific coasts of South America with a history of significant commercial exploitation. We aimed to evaluate the population genetic structure and the evolutionary history of South American sea lion along its distribution by analyses of mitochondrial DNA (mtDNA) and 10 nuclear microsatellites loci. We analyzed 147 sequences of mtDNA control region and genotyped 111 individuals of South American sea lion for 10 microsatellite loci, representing six populations (Peru, Northern Chile, Southern Chile, Uruguay (Brazil), Argentina and Falkland (Malvinas) Islands) and covering the entire distribution of the species. The mtDNA phylogeny shows that haplotypes from the two oceans comprise two very divergent clades as observed in previous studies, suggesting a long period (>1 million years) of low inter-oceanic female gene flow. Bayesian analysis of bi-parental genetic diversity supports significant (but less pronounced than mitochondrial) genetic structure between Pacific and Atlantic populations, although also suggested some inter-oceanic gene flow mediated by males. Higher male migration rates were found in the intra-oceanic population comparisons, supporting very high female philopatry in the species. Demographic analyses showed that populations from both oceans went through a large population expansion ~10,000 years ago, suggesting a very similar influence of historical environmental factors, such as the last glacial cycle, on both regions. Our results support the proposition that the Pacific and Atlantic populations of the South American sea lion should be considered distinct evolutionarily significant units, with at least two managements units in each ocean.