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Total mercury ([THg]) and selenium ([TSe]) concentrations were determined in California sea lion (Zalophus californianus) lanugo from the Gulf of California in 2021 and 2022. Relationships with sex, morphometrics, and year were evaluated. Following toxicological thresholds of concern for piscivorous mammals, most pups had a [THg] < 10 ppm, one pup (2021) had a [THg] > 20 ppm, no pups had a [THg] > 30 ppm. Females had significantly higher [TSe] than males; sex did not influence [THg]. [THg] and [TSe] in 2022 were significantly higher in the general population and male cohorts compared to 2021. Significant negative correlations were observed between [THg], [TSe], and morphometrics (2021). These results indicate that, compared to other pinniped species, regional California sea lions may have a decreased likelihood of experiencing Hg-related adverse health effects. Year-related changes in element concentrations suggest continued monitoring of this population to assess pinniped, environmental, and potentially, human health.

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Trace metals concentrations along with stable isotopes ratios were measured in marine algae, sea grass, sponges, echinoderms, mollusks, crustaceans, fishes, and the California sea lion, to assess the bioaccumulation potential and detect potential risks for top predators off Bahia Magdalena, Mexico. We assessed the trophic magnification factor (TMF) to determine the potential for biomagnification of 11 trace metals. The concentrations of Fe and Zn were one order of magnitude higher than all other metals. Concentrations of As, Cu, Cd, Co, Cr, Fe, Mn and Ni correlated negatively with trophic level, supporting trophic dilution (TMF < 1, p > 0.05), while Zn and Hg had significant trophic magnification (TMF > 1, p < 0.05) when assessing only the benthic-pelagic foodweb. This research provides a baseline concentration of metals in multiple species, metal-specific foodweb bioaccumulation and biomagnification of mercury, underscoring the key role of the macrobenthic community as biovectors for trophic transfer of Hg through the foodweb to the California sea lion.

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Microplastics (MPs, < 5 mm in size) are highly bioavailable to many taxa within the marine ecosystem, either ingested directly or indirectly through trophic transfer from polluted prey. The ingestion analysis of these MPs from top predators, such as pinnipeds in Mexico, is relatively unexplored. Forty-eight scats from California sea lions were collected on six rookeries along the Gulf of California. From these scat samples, 294 suspected MPs particles were classified and chemically analyzed; 34% were synthetic and semi-synthetic, and 66% were non-synthetic. Blue-colored polyethylene terephthalate fibers were the most common type of MP registered. During laboratory work, multiple contamination control measures were implemented. Although the ingestion pathway is still unknown, our results support the other authors that suggest the potential trophic transfer of MPs to top predators and incidental ingestion while foraging. The particles documented here provide important baseline information for future MP research in the Gulf of California.

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Background: The gastrointestinal (GI) bacterial communities of sea lions described to date have occasionally revealed large intraspecific variability, which may originate from several factors including different methodological approaches. Indeed, GI bacterial community surveys commonly rely on the use of a single hypervariable region (HR) of 16S rRNA, which may result in misleading structural interpretations and limit comparisons among studies. Here, we considered a multi-locus analysis by targeting six HRs of 16S rRNA with the aims of (i) comprehensively assessing the GI bacterial consortium in rectal samples from Zalophus californianus pups and (ii) elucidating structural variations among the tested HRs. In addition, we evaluated which HRs may be most suitable for identifying intrinsic, structurally related microbiome characteristics, such as geographic variations or functional capabilities. Methods: We employed a Short MUltiple Regions Framework (SMURF) approach using the Ion 16S™ Metagenomic Kit. This kit provides different proprietary primers designed to target six HRs of the 16S rRNA gene. To date, the only analytical pipeline available for this kit is the Ion Reporter™ Software of Thermo Fisher Scientific. Therefore, we propose an in-house pipeline to use with open-access tools, such as QIIME2 and PICRUSt 2, in downstream bioinformatic analyses. Results: As hypothesized, distinctive bacterial community profiles were observed for each analyzed HR. A higher number of bacterial taxa were detected with the V3 and V6-V7 regions. Conversely, the V8 and V9 regions were less informative, as we detected a lower number of taxa. The synergistic information of these HRs suggests that the GI microbiota of Zalophus californianus pups is predominated by five bacterial phyla: Proteobacteria (~50%), Bacteroidetes (~20%), Firmicutes (~18%), Fusobacteria (~7%), and Epsilonbacteraeota (~4%). Notably, our results differ at times from previously reported abundance profiles, which may promote re-evaluations of the GI bacterial compositions in sea lions and other pinniped species that have been reported to date. Moreover, consistent geographic differences were observed only with the V3, V4, and V6-V7 regions. In addition, these HRs also presented higher numbers of predicted molecular pathways, although no significant functional changes were apparent. Together, our results suggests that multi-locus analysis should be encouraged in GI microbial surveys, as single-locus approaches may result in misleading structural results that hamper the identification of structurally related microbiome features.

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Bacterial infections have been documented in marine mammals for decades, and some are considered emerging pathogens with zoonotic potential. The aerobic oral (n=16) and rectal (n=17) bacterial microbiota and their antimicrobial resistance were characterized for 17 apparently healthy California sea lion pups (Zalophus californianus) captured with a hoop net in Farallon Island, Sinaloa, Mexico, in 2016. Bacteriologic cultures, Analytical Profile Index, and PCR were used to identify bacterial species. The Escherichia coli phylogenetic groups were identified by PCR, Salmonella serotypes were identified, and resistance to antibiotics was evaluated. Overall, 39 bacterial species were isolated, including E. coli and Salmonella spp. (35.9% each) and Pseudomonas aeruginosa (28.2%). For E. coli, UNKNOWN phylogroup was the most prevalent (57.7%), followed by the A phylogroup (37.1%). Most Salmonella serotypes were identified as Newport (92.8%); serotype Saintpaul was also identified (7.2%). Sea lions with bacterial co-colonization included 24.2%, from which two bacterial species were isolated, and 3% with three species. Overall, 59% of bacteria were resistant to at least one antibiotic tested, and 25.6% were extensively drug resistant. Bacteria were highly resistant to ampicillin and cefotaxime. This study demonstrates the importance of characterizing the microbiome of sea lions, and the potential effect of pathogens with antimicrobial resistance on wildlife conservation and public health.

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BACKGROUND: The systematic of several marine diphyllobothriid tapeworms of pinnipeds has been revised in recent years. However, 20 species of Diphyllobothrium from phocids and otariids are still recognized as incertae sedis. We describe a new species of Diphyllobothrium from the intestine of California sea lions Zalophus californianus (Lesson) (type-host) and South American sea lions Otaria flavescens (Shaw). METHODS: Zalophus californianus from the Pacific coast of the USA and O. flavescens from Peru and Argentina were screened for parasites. Partial fragments of the large ribosomal subunit gene (lsrDNA) and the cytochrome c oxidase subunit 1 (cox1) mitochondrial gene were amplified for 22 isolates. Properly fixed material from California sea lions was examined using light and scanning electron microscopy. RESULTS: A total of four lsrDNA and 21 cox1 sequences were generated and aligned with published sequences of other diphyllobothriid taxa. Based on cox1 sequences, four diphyllobothriid tapeworms from O. flavescens in Peru were found to be conspecific with Adenocephalus pacificus Nybelin, 1931. The other newly generated sequences fall into a well-supported clade with sequences of a putative new species previously identified as Diphyllobothrium sp. 1. from Z. californianus and O. flavescens. A new species, Diphyllobothrium sprakeri n. sp., is proposed for tapeworms of this clade. CONCLUSIONS: Diphyllobothrium sprakeri n. sp. is the first diphyllobothriid species described from Z. californianus from the Pacific coast of North America, but O. flavescens from Argentina, Chile and Peru was confirmed as an additional host. The present study molecularly confirmed the first coinfection of two diphyllobothriid species in sea lions from the Southern Hemisphere.

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At least two species of filarial worms, Dirofilaria immitis and Acanthocheilonema (Dipetalonema) odendhali, infect otariid pinnipeds, including the California sea lion (Zalophus californianus). To date, evidence of infection in sea lions has come from dead or captive animals, and little is known about filariasis in free-living populations. We sampled 45 California sea lion adults and 197 pups captured at 12 rookeries from different ecological regions within the Gulf of California and detected and quantified D. immitis and A. odendhali microfilariae in blood smears. We investigated differences in prevalence and parasite load (intensity of infection) among ecological regions. Microfilariae were detected in the blood of 35 of the 45 (77.78%) adult females and in 1 of the 197 (0.51%) pups examined. The average burden of A. odendhali per microlitre of blood was nearly twice that of D. immitis. Prevalence and intensity of infection differed significantly among regions, being highest for colonies within the northern and northcentral regions and lowest in the southern region. Dirofilaria immitis and A. odendhali infections displayed a similar spatial pattern of prevalence. Colony density inversely predicted the prevalence of microfilariae. Based on the clinical parameters typically associated with filarial infections in carnivores and physical examinations, none of the sea lions appeared to have evidence of disease. This is a first approximation to investigate the prevalence of microfilaria infections in free-ranging California sea lions and to explore their relevance to population health.

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Introduction: The life history of the California sea lion (Zalophus californianus) in the Gulf of California is marked by a series of important events influencing and modifying its population growth, distribution, and evolution. Despite the fact that this population has been studied since the 1950s, research has been rather punctual and fragmentary. Before 2010, there are only a few surveys conducted simultaneously in all rookeries, thus there is no reliable information on key aspects of life cycle, population trend and potential threats. In the present work we conducted a review of California sea lion life history and environmental changes in the Gulf of California thorough a collation survey data encompassing the last 37 years. Objective: Our aim was focused on identifying short- and long-term processes potentially acting on the population, and hopefully improve knowledge about the population trend and status using different points of view. Methods: We collected and analyzed population survey data from different sources since the 1970s to 2018: published papers, master's and doctoral thesis, in addition to technical reports. The survey data are organized in sections corresponding with crucial population life history events. Results: Considering a long-time period the population size appears to be stable with zero growth. Cyclic interannual fluctuation seem to denote a certain dependence with climatic factors, not directly with El Niño, but with sea surface temperature anomalies that determine prey availability. However, many doubts persist about the incidence of different local environmental factors on gender and age, particularly related with juvenile recruitment and female survival rate. Conclusions: In conclusion, more information is required based on seasonal surveys, life cycle, regional environmental variation. Statistical errors need to be assessed and monitoring methods should be standardized and must be considered to ascertain short- and long-term population and colony spatial-temporal patterns.

Introducción: El lobo marino de California (Zalophus californianus) en el Golfo de California se caracteriza por una serie de eventos que influyen en el crecimiento, evolución y distribución de la población. Los estudios poblacionales iniciaron en 1950, aunque las investigaciones fueron puntuales y fragmentadas. Antes de 2010 existen pocos censos simultáneos de las 13 colonias de lobos marinos, con los cuales se obtuvo información sobre el ciclo de vida, la tendencia poblacional y las potenciales amenazas de estos organismos. Objetivo: En esta investigación se presenta un resumen de 37 años de historia de investigaciones del lobo marino de California y del ecosistema del Golfo de California. Métodos: Se realizó un análisis de los procesos de corto y largo plazo que actúan sobre la población, revelando aspectos poco conocidos. Se recolectaron y analizaron datos para el periodo de 1970 al 2018: artículos, tesis de posgrado y reportes técnicos, que fueron organizados en secciones relacionadas con eventos ambientales cruciales para la población. Resultados: A largo plazo, la población parece estar estable y presenta fluctuaciones interanuales cíclicas que evidencian dependencia con factores climáticos como anomalías de temperatura superficial del mar regional que determinan un cambio en la disponibilidad de presas para los lobos marinos. No es claro el posible efecto que factores ambientales locales puedan ocasionar en las diferentes clases de edad y por sexos, en particular sobre el reclutamiento de juveniles y la tasa de supervivencia de las hembras. Conclusiones: El presente trabajo identifica las prioridades de información para esta población y ofrece recomendaciones como el monitoreo estandarizado y la consideración de variaciones espacio-temporales locales.

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Ectopic pregnancies have been rarely reported in animals and little is known in wildlife species regarding the capacity of the placenta to support extrauterine fetal development. An adult, female, free-ranging California sea lion (Zalophus californianus) died suddenly after four weeks in a rehabilitation center. At necropsy, a partially mummified fetus, covered by light-brown membranes occupied the lower third of the abdominal cavity. The maternal uterus was intact and a prominent corpus luteus enlarged the right ovary. The fetal membranes epithelium had immunohistochemical profile identical to normal sea lion placenta but was different from the staining profile of sea lion omentum. The findings in this case suggest that partial placentation occurred in the abdominal cavity, highlighting the potential of otariids placenta to support ectopic fetal development. Acute toxic shock with disseminated intravascular coagulation was considered the most likely cause of death of the adult female sea lion.(AU)

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Ectopic pregnancies have been rarely reported in animals and little is known in wildlife species regarding the capacity of the placenta to support extrauterine fetal development. An adult, female, free-ranging California sea lion (Zalophus californianus) died suddenly after four weeks in a rehabilitation center. At necropsy, a partially mummified fetus, covered by light-brown membranes occupied the lower third of the abdominal cavity. The maternal uterus was intact and a prominent corpus luteus enlarged the right ovary. The fetal membranes epithelium had immunohistochemical profile identical to normal sea lion placenta but was different from the staining profile of sea lion omentum. The findings in this case suggest that partial placentation occurred in the abdominal cavity, highlighting the potential of otariids placenta to support ectopic fetal development. Acute toxic shock with disseminated intravascular coagulation was considered the most likely cause of death of the adult female sea lion.

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Inflammation is one of the most important non-specific and rapid responses that a vertebrate can elicit in response to damage or a foreign insult. To date, despite increasing evidence that the innate and adaptive branches of immunity are more intricately related than previously thought, few have examined interactions between the Major Histocompatibility Complex (MHC, a polymorphic region of the vertebrate genome that is involved with antigen presentation) and inflammation, and even less is known about these interactions in an eco-immunological context. Here, we examined the effect of MHC class II DRB gene multiplicity and transcription on phytohemagglutinin (PHA)-induced inflammation during the early stages of development of California sea lions. Neither constitutive nor expressed ZacaDRB diversity was found to be associated with pup responses to PHA at any of the stages of pup development. However, for two-month-old pups, those with a specific MHC-DRB locus (ZacaDRB-A) tended to have less efficient responsive inflammation. Transcription of distinct MHC-DRB loci was also linked to PHA-induced inflammation, with patterns that varied markedly between ages, and that suggested that ongoing infectious processes could limit the capacity to respond to a secondary challenge. Life history constraints and physiological processes associated with development of California sea lions, in conjunction with their changing pathogenic environment could explain the observed effects of MHC class II transcription on PHA-induced inflammation. To our knowledge, ours is the first study to examine the importance of expressed vs. constitutive MHC loci on inflammation in a natural population.

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The California sea lion ( Zalophus californianus ), a permanent inhabitant of the Gulf of California in Mexico, is susceptible to pathogenic Leptospira spp. infection, which can result in hepatic and renal damage and may lead to renal failure and death. During summer 2013, we used the microscopic agglutination test (MAT) to investigate the prevalence of anti-Leptospira antibodies in blood of clinically healthy sea lion pups from seven rookery islands on the Pacific Coast of Baja California (Pacific Ocean) and in the Gulf of California. We also used PCR to examine blood for Leptospira DNA. Isolation of Leptospira in liquid media was unsuccessful. We found higher antibody prevalence in sea lions from the rookery islands in the gulf than in those from the Pacific Coast. Antibodies against 11 serovars were identified in the Gulf of California population; the most frequent reactions were against serovars Bataviae (90%), Pyrogenes (86%), Wolffi (86%), Celledoni (71%), and Pomona (65%). In the Pacific Ocean population, MAT was positive against eight serovars, where Wolffi (88%), Pomona (75%), and Bataviae (70%) were the most frequent. Serum samples agglutinated with more than one Leptospira serovar. The maximum titer was 3,200. Each island had a different serology profile, and islands combined showed a distinct profile for each region. We detected pathogenic Leptospira DNA in 63% of blood samples, but we found no saprophytic Leptospira. Positive PCR results were obtained in blood samples with high and low MAT titers. Together, these two methods enhance the diagnosis and interpretation of sea lion leptospirosis. Our results may be related to human activities or the presence of other reservoirs with which sea lions interact, and they may also be related to sea lion stranding.

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