Detection of Leptospira kirschneri in a short-beaked common dolphin (Delphinus delphis delphis) stranded off the coast of southern California, USA.

BACKGROUND: Pathogenic Leptospira species are globally important zoonotic pathogens capable of infecting a wide range of host species. In marine mammals, reports of Leptospira have predominantly been in pinnipeds, with isolated reports of infections in cetaceans. CASE PRESENTATION: On 28 June 2021, a 150.5 cm long female, short-beaked common dolphin (Delphinus delphis delphis) stranded alive on the coast of southern California and subsequently died. Gross necropsy revealed multifocal cortical pallor within the reniculi of the kidney, and lymphoplasmacytic tubulointerstitial nephritis was observed histologically. Immunohistochemistry confirmed Leptospira infection, and PCR followed by lfb1 gene amplicon sequencing suggested that the infecting organism was L.kirschneri. Leptospira DNA capture and enrichment allowed for whole-genome sequencing to be conducted. Phylogenetic analyses confirmed the causative agent was a previously undescribed, divergent lineage of L.kirschneri. CONCLUSIONS: We report the first detection of pathogenic Leptospira in a short-beaked common dolphin, and the first detection in any cetacean in the northeastern Pacific Ocean. Renal lesions were consistent with leptospirosis in other host species, including marine mammals, and were the most significant lesions detected overall, suggesting leptospirosis as the likely cause of death. We identified the cause of the infection as L.kirschneri, a species detected only once before in a marine mammal - a northern elephant seal (Mirounga angustirostris) of the northeastern Pacific. These findings raise questions about the mechanism of transmission, given the obligate marine lifestyle of cetaceans (in contrast to pinnipeds, which spend time on land) and the commonly accepted view that Leptospira are quickly killed by salt water. They also raise important questions regarding the source of infection, and whether it arose from transmission among marine mammals or from terrestrial-to-marine spillover. Moving forward, surveillance and sampling must be expanded to better understand the extent to which Leptospira infections occur in the marine ecosystem and possible epidemiological linkages between and among marine and terrestrial host species. Generating Leptospira genomes from different host species will yield crucial information about possible transmission links, and our study highlights the power of new techniques such as DNA enrichment to illuminate the complex ecology of this important zoonotic pathogen.

Systemic Erysipelas Outbreak among Free-Ranging Bottlenose Dolphins, San Diego, California, USA, 2022.

We diagnosed fatal Erysipelothrix rhusiopathiae sepsis in 3 stranded bottlenose dolphins (Tursiops truncatus) during summer 2022, in San Diego, California, USA. The previously undetected disease in this relatively small, regional population of dolphins most likely indicates an environmental or biological change in the coastal ocean or organisms.

Assessing Marine Endocrine-Disrupting Chemicals in the Critically Endangered California Condor: Implications for Reintroduction to Coastal Environments.

Coastal reintroduction sites for California condors (Gymnogyps californianus) can lead to elevated halogenated organic compound (HOC) exposure and potential health impacts due to the consumption of scavenged marine mammals. Using nontargeted analysis based on comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GC×GC/TOF-MS), we compared HOC profiles of plasma from inland and coastal scavenging California condors from the state of California (CA), and marine mammal blubber from CA and the Gulf of California off Baja California (BC), Mexico. We detected more HOCs in coastal condors (32 ± 5, mean number of HOCs ± SD, n = 7) than in inland condors (8 ± 1, n = 10) and in CA marine mammals (136 ± 87, n = 25) than in BC marine mammals (55 ± 46, n = 8). ∑DDT-related compounds, ∑PCBs, and total tris(chlorophenyl)methane (∑TCPM) were, respectively, ∼7, ∼3.5, and ∼148 times more abundant in CA than in BC marine mammals. The endocrine-disrupting potential of selected polychlorinated biphenyls (PCB) congeners, TCPM, and TCPMOH was determined by in vitro California condor estrogen receptor (ER) activation. The higher levels of HOCs in coastal condors compared to those in inland condors and lower levels of HOC contamination in Baja California marine mammals compared to those from the state of California are factors to consider in condor reintroduction efforts.

Cooperative feeding in common dolphins as suggested by ontogenetic patterns in δ15 N bulk and amino acids.

Understanding the effect of stage-specific traits on species feeding habits can reveal how natural selection shapes life strategies. Amino acid (AA) nitrogen stable isotopes (δ15 N) provide multiple proxies of habitat baseline values and diet that can improve our understanding of species feeding strategies relative to their animal metabolism. We evaluated the effect of body length as a proxy for life stage and sex on the feeding habits of the common dolphin Delphinus delphis delphis using δ13 C and δ15 N in bulk tissue and AAs δ15 N from skin samples collected for almost two decades. For bulk δ13 C and δ15 N data, we used SIBER analysis to compare isotopic niches by sex and life stage. For AA δ15 N data, we developed a hierarchical Bayesian model (HBM) to estimate indices of trophic status (Δ15 N and trophic position). The model reflected the natural hierarchical structure of AA data by partitioning variability into three sources: between laboratory replicates, within dolphins and among dolphins. Estimates of Δ15 N based on all trophic and source AAs were more precise for each dolphin, less variable among dolphins and on average 2.4‰ higher than indices based on single trophic (Glx) and source (Phe) AAs. Precision was further increased when information was shared among individuals through random effects or regression models. Estimates of trophic position showed similar patterns. Both Δ15 N and δ15 Nbulk isotopic niches showed no difference by sex, suggesting that males and females have similar feeding habits and may not segregate. However, lower Δ15 N values for weaning calves and smaller juveniles discriminate them from adults, whereas δ15 N bulk isotopic niches do not. A trophic discrimination factor (TDFTro-Src ) of 3.1‰ was required for reasonable estimates of trophic position for these dolphins. Together, the lack of δ15 N differences between sexes, low variation between juveniles and adults and knowledge of common dolphins' social organization support intraspecific feeding cooperation as an important strategy to feed in the highly dynamic marine environment. Our study also presents an efficient way to analyse complex AA δ15 N data using HBM to investigate foraging behaviour in long-lived marine species difficult to study in the wild.

Renal and vaginal calculi in a free-ranging long-beaked common dolphin Delphinus capensis.

Bilateral nephrolithiasis with a concurrent vaginal calculus was identified in a stranded free-ranging long-beaked common dolphin Delphinus capensis. Necropsy and radiologic examinations of the sexually mature D. capensis revealed multiple small irregularly round nephroliths and a 6.4 × 4.1 × 9.2 cm vaginal calculus weighing 182 g. Nephroliths numbered 68 and 71 in the left and right kidneys, respectively, and ranged from 1.7 to 6.9 mm in diameter. Nephroliths were composed of 100% ammonium urate, which has been found in captive dolphin populations. However, the vaginolith consisted of struvite and calcium carbonate suggesting an alternate etiology. The composition of the vaginolith suggests that bacterial vaginitis could have served as the predisposing condition. Renal lesions included chronic tubulointerstitial nephritis with tubular degeneration and loss, likely secondary to the nephroliths. The pathogenesis of ammonium urate nephrolithiasis in managed care is suspected to be linked to diet and age but in this case may be due to metabolic disruption. However, if environmental changes cause a shift in prey species, the risk of nephrolithiasis in free-ranging cetaceans could increase. Careful surveillance for nephroliths in free-ranging populations should be considered by researchers.

Accuracy and precision of age determination using growth layer groups for California sea lions (Zalophus californianus) with known ages.

Age determination from counts of growth layer groups (GLGs) in tooth dentine is a common method for aging marine mammals. Using known-aged animals, we validated this method for acid etched teeth of California sea lions (CSLs), Zalophus californianus. Between 1991 and 2013, the upper left canine (n = 33) was collected opportunistically during necropsy from animals tagged or branded as pups that later died. Overall, 55%-61% of age estimates by GLG counting were within 1 yr of the known-age in the sample of 1-30-yr-old CSLs. Accuracy of age estimates was found to be dependent on age of the CSLs, however. 71%-79% of age estimates were within 1 yr of the known-age in CSLs <10 yr old. These findings support the validity of counting GLGs to estimate age for CSLs <10 yr old to within 1 yr of accuracy.

Newly Identified DDT-Related Compounds Accumulating in Southern California Bottlenose Dolphins.

Nontargeted GC×GC-TOF/MS analysis of blubber from 8 common bottlenose dolphins (Tursiops truncatus) inhabiting the Southern California Bight was performed to identify novel, bioaccumulative DDT-related compounds and to determine their abundance relative to the commonly studied DDT-related compounds. We identified 45 bioaccumulative DDT-related compounds of which the majority (80%) is not typically monitored in environmental media. Identified compounds include transformation products, technical mixture impurities such as tris(chlorophenyl)methane (TCPM), the presumed TCPM metabolite tris(chlorophenyl)methanol (TCPMOH), and structurally related compounds with unknown sources, such as hexa- to octachlorinated diphenylethene. To investigate impurities in pesticide mixtures as possible sources of these compounds, we analyzed technical DDT, the primary source of historical contamination in the region, and technical Dicofol, a current use pesticide that contains DDT-related compounds. The technical mixtures contained only 33% of the compounds identified in the blubber, suggesting that transformation products contribute to the majority of the load of DDT-related contaminants in these sentinels of ocean health. Quantitative analysis revealed that TCPM was the second most abundant compound class detected in the blubber, following DDE, and TCPMOH loads were greater than DDT. QSPR estimates verified 4,4',4″-TCPM and 4,4'4,″-TCPMOH are persistent and bioaccumulative.

Nontargeted biomonitoring of halogenated organic compounds in two ecotypes of bottlenose dolphins (Tursiops truncatus) from the Southern California Bight.

Targeted environmental monitoring reveals contamination by known chemicals, but may exclude potentially pervasive but unknown compounds. Marine mammals are sentinels of persistent and bioaccumulative contaminants due to their longevity and high trophic position. Using nontargeted analysis, we constructed a mass spectral library of 327 persistent and bioaccumulative compounds identified in blubber from two ecotypes of common bottlenose dolphins (Tursiops truncatus) sampled in the Southern California Bight. This library of halogenated organic compounds (HOCs) consisted of 180 anthropogenic contaminants, 41 natural products, 4 with mixed sources, 8 with unknown sources, and 94 with partial structural characterization and unknown sources. The abundance of compounds whose structures could not be fully elucidated highlights the prevalence of undiscovered HOCs accumulating in marine food webs. Eighty-six percent of the identified compounds are not currently monitored, including 133 known anthropogenic chemicals. Compounds related to dichlorodiphenyltrichloroethane (DDT) were the most abundant. Natural products were, in some cases, detected at abundances similar to anthropogenic compounds. The profile of naturally occurring HOCs differed between ecotypes, suggesting more abundant offshore sources of these compounds. This nontargeted analytical framework provided a comprehensive list of HOCs that may be characteristic of the region, and its application within monitoring surveys may suggest new chemicals for evaluation.

Clostridium perfringens septicemia in a long-beaked common dolphin Delphinus capensis: an etiology of gas bubble accumulation in cetaceans.

An adult female long-beaked common dolphin Delphinus capensis live-stranded in La Jolla, California, USA, on July 30, 2012 and subsequently died on the beach. Computed tomography and magnetic resonance imaging revealed gas bubble accumulation in the vasculature, organ parenchyma, mandibular fat pads, and subdermal sheath as well as a gas-filled cavity within the liver, mild caudal abdominal effusion, and fluid in the uterus. Gross examination confirmed these findings and also identified mild ulcerations on the palate, ventral skin, and flukes, uterine necrosis, and multifocal parenchymal cavitations in the brain. Histological review demonstrated necrosis and round clear spaces interpreted as gas bubbles with associated bacterial rods within the brain, liver, spleen, and lymph nodes. Anaerobic cultures of the lung, spleen, liver, bone marrow, and abdominal fluid yielded Clostridium perfringens, which was further identified as type A via a multiplex PCR assay. The gas composition of sampled bubbles was typical of putrefaction gases, which is consistent with the by-products of C. perfringens, a gas-producing bacterium. Gas bubble formation in marine mammals due to barotrauma, and peri- or postmortem off-gassing of supersaturated tissues and blood has been previously described. This case study concluded that a systemic infection of C. perfringens likely resulted in production of gas and toxins, causing tissue necrosis.

Gray whale (Eschrichtius robustus) post-mortem findings from December 2018 through 2021 during the Unusual Mortality Event in the Eastern North Pacific.

Beginning in December 2018, increased numbers of gray whale (Eschrichtius robustus) strandings were reported along the west coast of Mexico, the United States, and Canada, prompting declaration of a gray whale Unusual Mortality Event (UME) by the United States National Marine Fisheries Service. Although strandings declined in 2020 and 2021 from a peak in 2019, the UME is still ongoing as of fall 2023. Between 17 December 2018 and 31 December 2021, 503 animals stranded along the west coast of North America, with 226 strandings in Mexico, 71 in California, 12 in Oregon, 56 in Washington, 21 in British Columbia, and 117 in Alaska. These included 187 males, 167 females, and 149 whales of undetermined sex; and 193 adults, 194 subadults, 40 calves, 1 fetus, and 75 whales of undetermined age class. We report on 61 of the 503 carcasses (12%) that had external and internal gross necropsy and/or histopathology data: of these 61 whales, findings that contributed to death were identified in 33 (54%) whales. Sixteen of the 61 (26%) were severely emaciated. Gross lesions of blunt force trauma consistent with vessel strike were identified in 11 of the 61 animals (18%), only two of which were emaciated. Two whales (3%) were entangled at time of death, and one died from entrapment. Signs of killer whale (Orcinus orca) interaction were documented in 19 of the 61 animals; five were deemed from recent interactions and three (5%) likely contributed to mortality. A specific cause of death could not be identified in 28 of 61 whales (46%). Additionally, logistical challenges and the advanced state of decomposition of most examined carcasses precluded detection of potential infectious or toxic causes of morbidity or mortality. Up to 2016, the eastern North Pacific population of gray whale population had generally been increasing since the cessation of historic whaling and a prior UME in 1999-2000. However, recent abundance and calf production estimates have declined, a trend that overlaps the current UME. The relative contributions of carrying capacity, environmental change, prey shifts, and infectious, toxic, and other processes to the increased gray whale mortalities have not yet been resolved. Nevertheless, the marked temporal increase in strandings, including findings of malnutrition in some of the whales, along with low calf production, likely represent consequences of complex and dynamic ecological interactions in the ocean impacting the population.

Marine algal toxins and their vectors in southern California cetaceans.

Published baseline data on biotoxin exposure in cetaceans is sparse but critical for interpreting mortality events as harmful algal blooms increase in frequency and duration. We present the first synthesis of domoic acid (DA), saxitoxin (STX), okadaic acid (OA), and microcystin detections in the feces and urine of stranded and bycaught southern California cetaceans, over an 18 year period (2001-2018), along with corresponding stomach content data. DA was detected in 13 out of 19 cetacean species, most often in harbor porpoise (Phocoena phocoena) (81.8%, n = 22) and long-beaked common dolphins (Delphinus delphis bairdii) (74%, n = 231). Maximum DA concentrations of 324,000 ng/g in feces and 271, 967 ng/ml in urine were observed in D. d. bairdii. DA was detected more frequently and at higher concentrations in male vs. female D. d. bairdii. Higher fecal DA concentrations in D. d. bairdii were associated with a greater proportion of northern anchovy (Engraulis mordax) in the diet, indicating it may be a primary vector of DA. Fecal DA concentrations for D. d. bairdii off Point Conception were greater than those from animals sampled off Los Angeles and San Diego counties, reflecting greater primary productivity and higher Pseudo-nitzschia spp. abundance in that region and a greater abundance of E. mordax in the diet. STX was detected at low levels (fecal max = 7.5 ng/g, urine max = 17 ng/ml) in 3.6% (n = 165) of individuals from 3 out of 11 species. The occurrence of E. mordax in 100% of the 3 examined stomachs suggests this species could be a primary vector of the detected STX. OA was detected in 2.4% of tested individuals (n = 85) at a maximum fecal concentration of 422.8 ng/g. Microcystin was detected in 14.3% (n = 7) of tested individuals with a maximum liver concentration of 96.8 ppb.

Genomic and anatomical comparisons of skin support independent adaptation to life in water by cetaceans and hippos.

The macroevolutionary transition from terra firma to obligatory inhabitance of the marine hydrosphere has occurred twice in the history of Mammalia: Cetacea and Sirenia. In the case of Cetacea (whales, dolphins, and porpoises), molecular phylogenies provide unambiguous evidence that fully aquatic cetaceans and semiaquatic hippopotamids (hippos) are each other's closest living relatives. Ancestral reconstructions suggest that some adaptations to the aquatic realm evolved in the common ancestor of Cetancodonta (Cetacea + Hippopotamidae). An alternative hypothesis is that these adaptations evolved independently in cetaceans and hippos. Here, we focus on the integumentary system and evaluate these hypotheses by integrating new histological data for cetaceans and hippos, the first genome-scale data for pygmy hippopotamus, and comprehensive genomic screens and molecular evolutionary analyses for protein-coding genes that have been inactivated in hippos and cetaceans. We identified eight skin-related genes that are inactivated in both cetaceans and hippos, including genes that are related to sebaceous glands, hair follicles, and epidermal differentiation. However, none of these genes exhibit inactivating mutations that are shared by cetaceans and hippos. Mean dates for the inactivation of skin genes in these two clades serve as proxies for phenotypic changes and suggest that hair reduction/loss, the loss of sebaceous glands, and changes to the keratinization program occurred ∼16 Ma earlier in cetaceans (∼46.5 Ma) than in hippos (∼30.5 Ma). These results, together with histological differences in the integument and prior analyses of oxygen isotopes from stem hippopotamids ("anthracotheres"), support the hypothesis that aquatic skin adaptations evolved independently in hippos and cetaceans.

Vaquitas (Phocoena sinus) continue to die from bycatch not pollutants.

BACKGROUND: The vaquita of Mexico is critically endangered, with a population less than 19 individuals in 2018. The population continues to decline and gillnet use continues in vaquita habitat. METHODS: Nine vaquita carcases were examined from 2016 to 2018 to establish cause of death. Concentrations of polychlorinated biphenyls (PCBs), DDTs and polybrominated diphenyl ethers (PBDEs) in blubber (n=3) were determined using gas chromatography/mass spectrometry and faeces tested for domoic acid and saxitoxin. RESULTS: Carcases were in good nutritional status and had lesions and full stomachs consistent with fisheries bycatch. PCB, DDT and PBDE concentrations ranged between 94 and 180 ng/g, 500 and 1200 ng/g and 97 and 210 ng/g lipid weight, respectively, which are low compared with other marine mammals. No saxitoxin or domoic acid was detected. CONCLUSION: These findings support the conclusion that bycatch is the primary source of vaquita population decline and emphasise the need for stronger measures to eliminate gillnets from the Upper Gulf of California, Mexico.

Apex marine predators and ocean health: Proactive screening of halogenated organic contaminants reveals ecosystem indicator species.

Despite decades-long bans on the production and use of certain chemicals, many halogenated organic compounds (HOCs) are persistent and can bioaccumulate in the marine environment with the potential to cause physiological harm to marine fauna. Highly lipid-rich tissue (e.g., marine mammal blubber) functions as a reservoir for HOCs, and selecting ideal indicator species is a priority for retrospective and proactive screening efforts. We selected five marine mammal species as possible indicators for the Southern California Bight (SCB) and applied a non-targeted analytical method paired with an automated data reduction strategy to catalog a broad range of known, known but unexpected, and unknown compounds in their blubber. A total of 194 HOCs were detected across the study species (n = 25 individuals), 81% of which are not routinely monitored, including 30 halogenated natural products and 45 compounds of unknown structure and origin. The cetacean species (long-beaked common dolphin, short-beaked common dolphin, and Risso's dolphin) averaged 128 HOCs, whereas pinnipeds (California sea lion and Pacific harbor seal) averaged 47 HOCs. We suspect this disparity can be attributed to differences in life history, foraging strategies, and/or enzyme-mediated metabolism. Our results support proposing (1) the long- and short-beaked common dolphin as apex marine predator sentinels for future and retrospective biomonitoring of the SCB ecosystem and (2) the use of non-targeted contaminant analyses to identify and prioritize emerging contaminants. The use of a sentinel marine species together with the non-targeted analytical approach will enable a proactive approach to environmental contaminant monitoring.

Temporal variation in pelagic food chain length in response to environmental change.

Climate variability alters nitrogen cycling, primary productivity, and dissolved oxygen concentration in marine ecosystems. We examined the role of this variability (as measured by six variables) on food chain length (FCL) in the California Current (CC) by reconstructing a time series of amino acid-specific δ15N values derived from common dolphins, an apex pelagic predator, and using two FCL proxies. Strong declines in FCL were observed after the 1997-1999 El Niño Southern Oscillation (ENSO) event. Bayesian models revealed longer FCLs under intermediate conditions for surface temperature, chlorophyll concentration, multivariate ENSO index, and total plankton volume but not for hypoxic depth and nitrate concentration. Our results challenge the prevalent paradigm that suggested long-term stability in the food web structure in the CC and, instead, reveal that pelagic food webs respond strongly to disturbances associated with ENSO events, local oceanography, and ongoing changes in climate.

Blubber cortisol: a potential tool for assessing stress response in free-ranging dolphins without effects due to sampling.

When paired with dart biopsying, quantifying cortisol in blubber tissue may provide an index of relative stress levels (i.e., activation of the hypothalamus-pituitary-adrenal axis) in free-ranging cetacean populations while minimizing the effects of the act of sampling. To validate this approach, cortisol was extracted from blubber samples collected from beach-stranded and bycaught short-beaked common dolphins using a modified blubber steroid isolation technique and measured via commercially available enzyme immunoassays. The measurements exhibited appropriate quality characteristics when analyzed via a bootstraped stepwise parallelism analysis (observed/expected = 1.03, 95%CI: 99.6 - 1.08) and showed no evidence of matrix interference with increasing sample size across typical biopsy tissue masses (75-150 mg; r(2) = 0.012, p = 0.78, slope = 0.022 ng(cortisol deviation)/ul(tissue extract added)). The relationships between blubber cortisol and eight potential cofactors namely, 1) fatality type (e.g., stranded or bycaught), 2) specimen condition (state of decomposition), 3) total body length, 4) sex, 5) sexual maturity state, 6) pregnancy status, 7) lactation state, and 8) adrenal mass, were assessed using a Bayesian generalized linear model averaging technique. Fatality type was the only factor correlated with blubber cortisol, and the magnitude of the effect size was substantial: beach-stranded individuals had on average 6.1-fold higher cortisol levels than those of bycaught individuals. Because of the difference in conditions surrounding these two fatality types, we interpret this relationship as evidence that blubber cortisol is indicative of stress response. We found no evidence of seasonal variation or a relationship between cortisol and the remaining cofactors.

Validation of blubber progesterone concentrations for pregnancy determination in three dolphin species and a porpoise.

Recent studies have validated the use of biopsies as a minimally invasive way to identify pregnant females in several species of wild cetaceans: Balaenapteraacutorostrata , Delphinusdelphis , Lissodelphisborealis, and Lagenorhynchusobliquidens. These studies found that progesterone (P4) concentrations quantified from blubber attached to biopsy samples is diagnostic of pregnancy. Here we examine a broader group of cetacean species in efforts to investigate how progesterone levels vary between species with respect to pregnancy status. We compared P4 concentrations in blubber collected from fishery bycatch and beach-stranded specimens for 40 females of known reproductive condition from Delphinuscapensis (n = 18), Stenellaattenuata (n = 8), S. longirostris (n = 6), and Phocoenoidesdalli (n = 8). The P4 concentrations were different (t = -7.1, p = 1.79E-08) between pregnant and non-pregnant animals in all species, with the mean blubber P4 concentration for pregnant animals 164 times higher than that of non-pregnant animals. There was no overlap in concentration levels between sexually immature or non-pregnant sexually mature animals and pregnant animals. No significant differences (F = 0.354, p = 0.559) were found between mature non-pregnant and immature D. capensis and Pdalli, suggesting P4 level is not indicative of maturity state in female delphinoids. P4 concentrations in relation to reproductive state were remarkably similar across species. All samples were analyzed with two different enzyme immunoassay kits to gauge assay sensitivity to measure progesterone in small samples, such as biopsies. With the technique now validated for these cetacean species, blubber P4 is a reliable diagnostic of pregnancies across multiple species, and thus expands the utility of this method to study reproduction in free-ranging cetaceans using biopsies.