Associations between total mercury, trace minerals, and blood health markers in Northwest Atlantic white sharks (Carcharodon carcharias).

The ecology and life-histories of white sharks make this species susceptible to mercury bioaccumulation; however, the health consequences of mercury exposure are understudied. We measured muscle and plasma total mercury (THg), health markers, and trace minerals in Northwest Atlantic white sharks. THg in muscle tissue averaged 10.0 mg/kg dry weight, while THg in blood plasma averaged 533 µg/L. THg levels in plasma and muscle were positively correlated with shark precaudal length (153-419 cm), and THg was bioaccumulated proportionally in muscle and plasma. Nine sharks had selenium:mercury molar ratios in blood plasma >1.0, indicating that for certain individuals the potential protective effects of the trace mineral were diminished, whereas excess selenium may have protected other individuals. No relationships between plasma THg and any trace minerals or health markers were identified. Thus, we found no evidence of negative effects of Hg bioaccumulation, even in sharks with very high THg.

Leucism in sharks: a histological examination.

Isolated cases of skin pigment disorders, including leucism, in sharks and rays have been reported for multiple species. Nonetheless, the morphological basis behind these chromatic anomalies has not been examined histologically. In this study, the authors quantified and compared the presence of melanin in multiple tissue samples of leucistic and fully pigmented blacktip sharks Carcharhinus limbatus. The authors' results support lack of melanin to be responsible for leucistic colouration. The histological differences responsible were evaluated.

Characterization of the complete mitochondrial genome of the brazilian cownose ray Rhinoptera brasiliensis (Myliobatiformes, Rhinopteridae) in the western Atlantic and its phylogenetic implications.

BACKGROUND: The Brazilian cownose ray, Rhinoptera brasiliensis has undergone a global population reduction and is currently classified by IUCN as Vulnerable. This species is sometimes confused with Rhinoptera bonasus, the only external diagnostic characteristic to distinguish between both species is the number of rows of tooth plates. Both cownose rays overlap geographically from Rio de Janeiro to the western North Atlantic. This calls for a more comprehensive phylogenetic assessment using mitochondria DNA genomes to better understand the relationships and delimitation of these two species. METHODS AND RESULTS: The mitochondrial genome sequences of R. brasiliensis was obtained by next-generation sequencing. The length of the mitochondrial genome was 17,759 bp containing 13 protein-coding genes (PCGs), two ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes, and a non-coding control region (D-loop). Each PCG was initiated by an authoritative ATG codon, except for COX1 initiated by a GTG codon. Most of the PCGs were terminated by a complete codon (TAA/TAG), while an incomplete termination codon (TA/T) was found in five out of the 13 PCGs. The phylogenetic analysis showed that R. brasiliensis was closely related to R. steindachneri whereas the reported mitogenome as R. steindachneri (GenBank accession number KM364982), differs from multiple mitocondrial DNA sequences of R. steindachneri and is nearly identical to that of R. javanica. CONCLUSION: The new mitogenome determined in this study provides new insight into the phylogenetic relationships in Rhinoptera, while providing new molecular data that can be applied to population genetic studies.

Legacy and emerging flame retardants in sharks from the Western North Atlantic Ocean.

Following the discontinuation of commercial polybrominated diphenyl ether (PBDE) mixtures, a variety of alternative flame retardants (FRs) have been developed and employed. To understand the contamination status of these emerging FRs in marine fish and wildlife, we investigated their bioaccumulation in four shark species, including shortfin mako shark (Isurus oxyrhinchus; n = 26), porbeagle (Lamna nasus; n = 4), sandbar shark (Carcharhinus plumbeus; n = 6), and common thresher (Alopias vulpinus; n = 4), from coastal and offshore waters of the western North Atlantic Ocean. Median concentrations of emerging FRs, including dechlorane analogues (i.e., dechlorane plus, Dec-602, -603, and - 604), tetrabromo-o-chlorotoluene (TBCT), 1,2-bis(2,4,6-tribromophenoxy)ethane (BTBPE), and hexabromobenzene (HBBZ), ranged from 1.4-7.4, 10.2-22.4, 1.0-16.7, and 4.1-17.7 ng/g lipid weight (lw), respectively. Although concentrations of emerging FRs were generally 1-2 orders of magnitude lower than those of legacy FRs (i.e., PBDEs, 312-800 ng/g lw and hexabromocyclododecane or HBCDD, 17.2-99.3 ng/g lw), they were detected in more than 80% of the shark livers. Analysis of available biological data indicated that fork length significantly correlated with the concentrations of ΣPBDEs, HBCDD, ΣDechloranes or TBCT in shortfin mako livers. This indicates that longer-term exposure likely results in elevated FR concentrations in sharks. Our findings suggest likely exposure of western North Atlantic fish and wildlife to several emerging FRs, including dechloranes, BTBPE, HBBZ, and TBCT. Additional studies are needed to better elucidate their potential risks to fish and wildlife as well as the variety of environmental and biological factors influencing these risks.

Characterization of Halogenated Organic Compounds in Pelagic Sharks and Sea Turtles Using a Nontargeted Approach.

Halogenated organic compounds (HOCs) in marine species collected from the Atlantic Ocean [3 shortfin mako (Isurus oxyrinchus) and 1 porbeagle (Lamna nasus)], and 12 sea turtles collected from the Pacific Ocean [3 loggerhead (Caretta caretta), 3 green (Chelonia mydas), 3 olive ridley (Lepidochelys olivacea), and 3 hawksbill (Eretmochelys imbricata)] were analyzed with a nontargeted analytical method using two-dimensional gas chromatography coupled to high-resolution time-of-flight mass spectrometry. Sharks and sea turtles had distinct HOC profiles. Halogenated methoxyphenols (halo-MeOPs) were the most abundant compound class identified in sea turtle livers, while polychlorinated biphenyls (PCBs) were the most abundant in shark livers. In addition to legacy contaminants and halo-MeOPs, a total of 110 nontargeted/novel HOCs (NHOCs) were observed in the shark livers. Shortfin mako collected from the northern Gulf of Mexico contained the largest number (89) and most diverse structural classes of NHOCs. Among all NHOCs, a group of compounds with the elemental composition C14H12-nCln (n = 5-8) exhibited the highest concentrations, followed by chlorocarbazoles and tris(chlorophenyl) methanes (TCPMs). Using nontargeted workflows, a variety of known and unknown HOCs were observed, which demonstrate the need to develop more complete chemical profiles in the marine environment.

Metal concentrations and metallothionein metal detoxification in blue sharks, Prionace glauca L. from the Western North Atlantic Ocean.

BACKGROUND: Elasmobranchs are particularly vulnerable to environmental metal contamination, accumulating these contaminants at high rates and excreting them slowly. The blue shark Prionace glauca L. is one of the most heavily fished elasmobranchs, although information regarding metal contamination and detoxification in this species is notably lacking. METHODS: Blue sharks were sampled in the western North Atlantic Ocean, in offshore waters adjacent to Cape Cod, Massachusetts. Total and metallothionein-bound liver and muscle metal concentrations were determined by inductively coupled plasma mass spectrometry (ICP-MS), metallothionein detoxification and oxidative stress endpoints were determined by UV-vis spectrophotometry. RESULTS: Metallothionein detoxification occurred for As, Cd, Cs, Cu, Hg, Pb, Se, Ti and Zn in liver, and for As, Cd, Cs, Pb, Se, and Zn in muscle, while reduced glutathione defenses seem to be related to Co and Zn exposure. CONCLUSION: This is the first report for several metals (Ag, Co, non-radioactive Cs, Sb, Ti and V) for this species, which will aid in establishing baseline elemental data for biomonitoring efforts, health metrics, and conservation measures.

Evaluation of muscle tissue as a non-lethal proxy for liver and brain organic contaminant loads in an elasmobranch, the Bonnethead Shark (Sphyrna tiburo).

Elasmobranch ecotoxicological investigations are complicated because accessing organs that accumulate organic contaminants is usually lethal. Several metrics among liver, muscle, and brain were evaluated to determine their relative organic contaminant loads and the efficacy of using muscle as a non-lethal proxy for liver. Liver contained the highest concentrations (368-4020 ng/g wet weigth [ww]) and greatest estimated total load of contaminants. Brain had higher toxin concentrations than muscle (4.18-84.2 ng/g ww versus 0.94-4.73 ng/g ww). Liver and brain were similar to each other in terms of contaminant detection occurrence and signature overlap, whereas muscle poorly reflected those of liver and brain. However, the identity of contaminants detected in muscle constituted those that substantially contributed to summed liver and brain concentrations. Thus, studies utilizing muscle as a non-lethal liver alternative to study organic contaminant exposure in elasmobranchs should craft questions with care, considering its limited ability to serve as an accurate proxy.

Mitochondrial DNA genome evidence for the existence of a third divergent lineage in the western Atlantic Ocean for the bull shark (Carcharhinus leucas).

We report for the first time a highly divergent lineage in the Caribbean Sea for the bull shark (Carcharhinus leucas) based on the analysis of 51 mitochondrial DNA genomes of individuals collected in the western North Atlantic. When comparing the mtDNA control region obtained from the mitogenomes to sequences reported previously for Brazil, the Caribbean lineage remained highly divergent. These results support the existence of a discrete population in Central America due to a phylogeographic break separating the Caribbean Sea from the western North Atlantic, Gulf of Mexico and South America.

High-resolution Spatiotemporal Dynamics of Harmful Algae in the Indian River Lagoon (Florida)-A Case Study of Aureoumbra lagunensis, Pyrodinium bahamense, and Pseudo-nitzschia.

The Indian River Lagoon (IRL), located on the east coast of Florida, is a complex estuarine ecosystem that is negatively affected by recurring harmful algal blooms (HABs) from distinct taxonomic/functional groups. Enhanced monitoring was established to facilitate rapid quantification of three recurrent bloom taxa, Aureoumbra lagunensis, Pyrodinium bahamense, and Pseudo-nitzschia spp., and included corroborating techniques to improve the identification of small-celled nanoplankton (<10 µm in diameter). Identification and enumeration of these target taxa were conducted during 2015-2020 using a combination of light microscopy and species-specific approaches, specifically immunofluorescence flow cytometry as well as a newly developed qPCR assay for A. lagunensis presented here for the first time. An annual bloom index (ABI) was established for each taxon based on occurrence and abundance data. Blooms of A. lagunensis (>2×108 cells L-1) were observed in all six years sampled and across multiple seasons. In contrast, abundance of P. bahamense, largely driven by the annual temperature cycle that moderates life cycle transitions and growth, displayed a strong seasonal pattern with blooms (105-107 cells L-1) generally developing in early summer and subsiding in autumn. However, P. bahamense bloom development was delayed and abundance was significantly lower in years and locations with sustained A. lagunensis blooms. Pseudo-nitzschia spp. were broadly distributed with sporadic bloom concentrations (reaching 107 cells L-1), but with minimal concentrations of the toxin domoic acid detected (<0.02 µg L-1). In summer 2020, multiple monitoring tools characterized a novel nano-cyanobacterium bloom (reaching 109 cells L-1) that coincided with a decline in A. lagunensis and persisted into autumn. Statistical and time-series analyses of this spatiotemporally intensive dataset highlight prominent patterns in variability for some taxa, but also identifies challenges of characterizing mechanisms underlying more episodic yet persistent events. Nevertheless, the intersect of temperature and salinity as environmental proxies proved to be informative in delineating niche partitioning, not only in the case of taxa with long-standing data sets but also for seemingly unprecedented blooms of novel nanoplanktonic taxa.

Histologic Observations of Dermal Wound Healing in a Free-Ranging Blacktip Shark from the Southeastern U.S. Atlantic Coast: A Case Report.

Despite indications that sharks have an exceptional capacity to heal from traumatic injuries, no detailed microscopic observations of integumental wounds have been reported for sharks. This study details the histopathological features of such wounds in a free-ranging shark. An adult male Blacktip Shark Carcharhinus limbatus was collected in 2017 during fisheries-independent sampling efforts in the coastal southeastern U.S. Atlantic. The shark had numerous lesions on his head, torso, and left pectoral fin that were compatible with shark bites. Representative samples from two wounds on the head were examined by light microscopy. The epidermal changes included hyperplasia and hypertrophy; intracellular edema; and the absence of goblet cells, denticles, and chromatophores. In the dermis, fibrinocellular exudation, granulation tissue, and marked skeletal muscle necrosis and regeneration were observed. The above features were comparable to wound healing in bony fish, albeit minor differences were found. Although this case documents exceptionally good regeneration of skeletal muscle in the shark, we found no evidence of unique morphological healing patterns. Further studies on wound healing are needed because recent molecular and genetic findings do suggest evolutionary adaptations enhancing healing in sharks.

Environmental management of two of the world's most endangered marine and terrestrial predators: Vaquita and cheetah.

Two of the world most endangered marine and terrestrial species are at the brink of extinction. The vaquita (Phocoena sinus) is the smallest existing cetacean and the population has declined to barely 22 individuals now remaining in Mexico's Gulf of California. With the ongoing decline, it is likely to go extinct within few years. The primary threat to this species has been mortality as a result of by-catch from gillnet fishing as well as environmental toxic chemicals and disturbance. This has called for the need to establish a National Park within the Gulf of California to expand essential habitat and provide the critical ecosystem protection for vaquita to thrive and multiply, given that proper conservation enforcement and management of the park are accomplished. In the terrestrial environment, the cheetah (Acinonyx jubatus) is reduced to a low number worldwide with the Iran subpopulation currently listed as Critically Endangered and the Indian subpopulation already extinct. There is a need for conservation efforts due to habitat loss, but also an indication of the conspicuous threat of illegal trade and trafficking from Africa and Arab countries in the Middle East. Funds have also been set up to provide refuges for the cheetah by working directly with farmers and landowners, which is a critical movement in adaptive management. These are the potential options for the preservation and possibly the expansion of the overall vaquita and cheetah populations.

Hybridization between sympatric hammerhead sharks in the western North Atlantic Ocean.

Hybridization between closely related species has been documented across a wide range of taxa but has not been well studied in elasmobranchs. Hammerhead sharks have drawn global conservation concern because they experience some of the highest mortality rates among sharks when interacting with fisheries. Here we report on the detection of hybrids between the globally distributed scalloped hammerhead ( Sphyrna lewini) and recently described Carolina hammerhead ( S. gilberti) which are only known from the western Atlantic Ocean. Using a genomics approach, 10 first-generation hybrids and 15-17 backcrosses were detected from 554 individuals. The identification of backcrosses demonstrates hybrids are viable, and all backcrosses but one involved a scalloped hammerhead. All hybrids but one possessed Carolina hammerhead mtDNA, indicating sex-biased gene flow between species. Repeated hybridization and backcrossing with scalloped hammerheads could lead to the loss of endemic Carolina hammerheads.

Correction to: Mercury accumulation, speciation, and temporal trends in Atlantic Stingrays (Hypanus sabinus).

The corrections are: In the 2nd sentence of the Statistical Analyses section, the current sentence lacks important statistical assumption wording.

Novel Dechlorane Analogues and Possible Sources in Peregrine Falcon Eggs and Shark Livers from the Western North Atlantic Regions.

During the investigation of dechlorane-related chemicals in North American wildlife, two unknown polychlorinated compounds (referred to as U1 and U2) were discovered. After extensive sample cleanup, structural information on U1 and U2 was characterized by gas chromatography (GC) coupled with single quadrupole mass spectrometer (MS) or GC-quadrupole time-of-flight (QToF) MS. Mass spectral evidence suggests that both U1 and U2 are structurally related to Dechlorane 603 (Dec603; C17H8Cl12), an analogue of the chlorinated flame retardant Dechlorane Plus. From the results we suspect U1 (C17H9Cl11) to be a monohydro analogue of Dec603 (i.e., one chlorine atom in Dec603 is replaced by a hydrogen atom). U1 may be formed via the degradation of Dec603's stereoisomers or present as an impurity in commercial Dec603 products. Mass spectral characterization of U2 (C17H7OCl11) suggests it is a carbonylic derivative of Dec603, likely formed via metabolic transformation of Dec603 or its photoisomer. Semiquantitative measurement revealed that U1 and U2 were present at estimated median concentrations of 49 ng/g lipid weight (lw) and 59 ng/g lw in peregrine falcon ( Falco peregrinus) eggs, from the mid-Atlantic region of the United States, and 4.6 and 3.0 ng/g lw in shortfin mako shark ( Isurus oxyrinchus) livers from the western North Atlantic Ocean, respectively. Our results demonstrate the occurrence of these two novel Dec603-related chemicals in both terrestrial and aquatic ecosystems.

Mercury accumulation, speciation, and temporal trends in Atlantic Stingrays (Hypanus sabinus).

Mercury (Hg) deposited into aquatic sediments can be converted into the more toxic methylmercury (MeHg) by microbial activity. Atlantic stingrays (Hypanus sabinus) are an estuarine and nearshore species found in coastal regions of the western North Atlantic, occurring in multiple habitat types, and feeding mainly on benthic invertebrates. Mercury dynamics and speciation in stingrays have not been well examined. This study quantified total Hg and Hg species (Hg (II) and MeHg) in Atlantic stingrays sampled from Florida's Indian River Lagoon (IRL) from 2012 to 2013. Tissues (muscle and liver) collected from 29 stingrays were lyophilized and homogenized before being analyzed using a direct mercury analyzer. Concentrations of total Hg in muscle were positively related to stingray disk width, but concentrations in liver were not. Mean (±SD) total Hg in muscle (0.56 ± 0.30 mg/kg dw) was significantly higher than mean total Hg in liver (0.23 ± 0.19 mg/kg dw). Within liver tissue, percent MeHg (of total Hg) ranged from 31 to 99%. The ratio between total Hg in liver and total Hg in muscle was <1 for nearly all individuals, suggesting a lack of active hepatic demethylation and sequestration mechanisms. Concentrations of Hg in IRL Atlantic stingrays fall below concentrations known to result in direct toxicity to fishes; however, effects thresholds are not well understood for elasmobranchs. Comparisons of Hg concentrations in IRL Atlantic stingrays sampled previously (37 individuals in 1994) indicate that total Hg concentrations in muscle of Atlantic stingrays have decreased over the past two decades, suggesting a reduction in the bioavailable Hg in the IRL ecosystem.

Maternal Transfer of Flame Retardants in Sharks from the Western North Atlantic Ocean.

The present work represents a comprehensive study of in utero maternal transfer of legacy and emerging flame retardants (FRs) in marine predators. We analyzed liver tissues from pregnant sharks of five viviparous species, including blacknose shark ( Carcharhinus acronotus; n = 12), blacktip shark ( Carcharhinus limbatus; n = 2), bonnethead ( Sphyrna tiburo; n = 2), Atlantic sharpnose shark ( Rhizoprionodon terraenovae; n = 2), and spinner shark ( Carcharhinus brevipinna; n = 2), as well as their embryos ( n = 84 in total from five species), collected from the western North Atlantic Ocean. Concentrations of frequently detected emerging FRs in adult female blacknose sharks were determined to be 6.1-83.3 ng/g lipid weight (lw) for dechlorane analogues, 2.5-29.8 ng/g lw for tetrabromo- o-chlorotoluene, and nondetection -32.6 ng/g lw for hexabromobenzene. These concentrations were 1-2 orders of magnitude lower than those of legacy polybrominated diphenyl ether flame retardants (85.7-398 ng/g lw). Similar contamination profiles were also found in the other four species, although FR concentrations varied in different species. A total of 21 FRs were commonly found in developing embryos of female sharks from five species, demonstrating maternal transfer in utero. The maternal transfer ratio (i.e., ratio of the mean litter concentration to their mother's concentration) determined in blacknose shark mother/embryo groups for each FR chemical was negatively associated with its octanol-water partition coefficient. Our work lays a solid foundation for future investigation of the underlying mechanisms of in utero transfer and additional physical or chemical factors that affect maternal transfer.

Perfluorinated alkyl acids and fecundity assessment in striped mullet (Mugil cephalus) at Merritt Island national wildlife refuge.

This study investigated wild caught striped mullet (Mugil cephalus) at Merritt Island National Wildlife Refuge (MINWR) for levels of 15 perfluoroalkyl acids (PFAA) in tandem with individual fecundity measurements (Oocyte sub-stage 2 late, n=42) and oocyte reproductive stages (Stages 1-5, n=128). PFAA measurements were quantified in striped mullet liver (n=128), muscle (n=49), and gonad (n=10). No significant negative impacts of liver PFAA burden on wild-caught, mullet fecundity endpoints were observed in this study; however, changes in PFAA were observed in the liver as mullet progressed through different sub-stages of oocyte development. Of the PFAA with significant changes by sub-stage of oocyte development, the carboxylic acids (perfluorooctanoic acid, perfluorononanoic acid, and perfluorotridecanoic acid) increased in the liver with increasing sub-stage while the sulfonic acid and its precursor (perfluorooctanesulfonic acid (PFOS) and perfluorooctanesulfonamide, respectively) decreased in the liver with increasing sub-stage of oocyte development. This is a unique find and suggests PFAA change location of compartmentalization as mullet progress towards spawning. Investigations also revealed higher than expected median muscle and gonad levels of PFOS in striped mullet collected at MINWR (9.01ng/g and 80.2ng/g, respectively).