Per-and poly-fluorinated alkyl substances (PFAS) are a group of chemicals used in a wide variety of commercial products and industrial applications. These chemicals are persistent, can accumulate in humans' and animals' tissues and in the environment, representing an increasing concern due to their moderate to highly toxicity. Their global distribution, persistence and toxicity led to an urgent need to investigate bioaccumulation also in marine species. In 2013 PFAS contamination was detected in a vast area in Veneto region, mainly in Adige and Brenta rivers. In order to investigate any relevant presence of these substances in marine vertebrates constantly living in the area, PFAS were measured in hepatic tissue samples of 20 bottlenose dolphins (Tursiops truncatus) stranded along the northern Adriatic Sea coastline between 2008 and 2020. Using high performance liquid chromatography-mass spectrometry, 17 target PFAS (PFBA, PFPeA, PFHxA, PFHpA, PFOA, PFNA, PFDA, PFUnA, PFDoA, PFTrDA, PFTeDA, PFBS, PFHxS, PFOS, PFDS, PFHpS, PFPeS), were quantified in the samples. PFAS profiles were generally composed of the same five dominant PFAS (PFOS > PFUnA > PFDA ≈ PFDoA ≈ PFTrDA). The greatest PFOS concentration found was 629,73 ng/g wet weight, and PFOS accounted until 71% in the PFAS profiles. No significant differences between sexes were found, while calves showing higher mean values than adults, possibly indicating an increasing ability in the elimination of PFAS with age. Finally, a temporal analysis was carried out considering three different periods of time, but no temporal differences in concentrations were found. The results suggest that long-chain PFAS are widespread in bottlenose dolphins along the North Adriatic Sea. Furthermore, they represent a baseline to investigate the impact of PFAS on marine mammals’ conservation and health. Filling an important gap in the knowledge of PFAS accumulation in bottlenose dolphins, this study highlights the relevant role of Environmental and Tissue Banks for retrospective analyses on emergent contaminants.

Odontocete cetaceans bioaccumulate high concentrations of endocrine disrupting persistent organic pollutants (POPs), including dichlorodiphenyltrichloroethane (DDT), dichlorodiphenyltrichloroethylene (DDE), and dichlorodiphenyldichloroethane (DDD) – collectively DDTs – but few studies have explored DDTs-mediated endocrine disruption in cetaceans. Herein, we use remotely collected blubber biopsies from common bottlenose dolphins (Tursiops truncatus) inhabiting a site with high localized DDTs contamination to study the relationships between DDTs exposure and steroid hormone homeostasis in cetaceans. We quantified blubber steroid hormone concentrations by liquid chromatography-tandem mass spectrometry and blubber POP concentrations by gas chromatography-mass spectrometry. We detected six steroid hormones in blubber, including progesterone (P4), 17-hydroxyprogesterone (17OHP4), androstenedione (AE), testosterone (T), cortisol (F), and cortisone (E). Sampled dolphins (n = 62) exhibited exposure to DDT, DDE, DDD, chlordanes (CHLDs), mirex, dieldrin, hexachlorobenzene, polychlorinated biphenyls (PCBs), and brominated diphenyl ethers (BDEs). Using principal components analysis (PCA), we determined that blubber DDTs primarily loaded to the first principal component (PC1) explaining 81.6% of the total variance in POP exposure, while the remaining POPs primarily loaded to the PC2 (10.4% of variance). PC1 scores were negatively correlated with blubber T in males and blubber F in females, suggesting that exposure to DDTs impacted androgen and corticosteroid homeostasis. These conclusions were further supported by observed negative correlations between T and o,p’-DDE, o,p’-DDD, and p,p’-DDD in males sampled in the fall, and between F and the six individual DDTs and ∑6DDTs in females. Overall, these results suggest that POP-mediated endocrine disruption may have occurred in this stock of dolphins, which could negatively impact their health and fitness. However, this study relied on uncontrolled incidental exposures, making it impossible to establish a causal relationship between DDTs exposure and endocrine effects. Importantly, this study demonstrates that remotely collected blubber biopsies are a useful matrix for studying endocrine disruption in marine mammals.

Dolphins are good bioindicators of the contamination status of marine ecosystems, since their dietary and habitat plasticity in both coastal and offshore ecotypes provide information on the trace elements levels originated from natural and anthropogenic sources. In this context, this study aimed to investigate provides mercury (Hg), selenium (Se) levels, trophic ecology and feeding environments of four small cetaceans (Tursiops truncatus, Steno bredanensis, Sotalia guianensis and Pontoporia blainvillei) inhabiting the central-northern coast of Rio de Janeiro State, southeastern Brazil. For the latter, δ15N and δ13C stable isotopes were used as indicators in this regard. Stable isotope values indicate that the four studied species have distinctive foraging habitats, coastal and least coastal, and occupy different trophic positions. The significant relationship found between muscle Hg and δ15N suggests that individual foraging preference remains relatively constant for the studied dolphin species over extended periods. The individual prey size and species are probably responsible for the differences found in Hg and Se concentrations in muscle tissue among all dolphin species. The vulnerable small coastal cetacean, P. blainvillei, which feeds on small teleost fish and squid, presented the lowest muscular Hg concentrations (less than 3.5 μg g−1 dry wt.). Meanwhile, S. bredanensis is more likely to uptake large amounts of trace elements among the four dolphin species, due to its feeding habits mainly being large offshore fish that accumulate high amounts of trace elements in organs and tissues. Differences found between Hg concentrations in fetus-mother pairs were much higher in S. guianensis than in P. blainvillei, suggesting that maternal contribution of Hg via placenta was more significant for the former.

Occurrence and behaviour of classical (PBDEs) and alternative (HNs, HBB, PBEB, DBDPE and HBCD) flame retardants, together with naturally produced MeO-PBDEs, were studied in short-beaked common dolphin (Delphinus delphis), bottlenose dolphin (Tursiops truncatus) and long-finned pilot whale (Globicephala melas) in two sampling locations from Southern European waters. PBDEs, Dec 602, Dec 603, DP, α-HBCD and two MeO-PBDEs were detected in all three species. ∑PBDEs were between 17 and 2680 ng/g lw; ∑HNs were between 1.1 and 59 ng/g lw; α-HBCD levels ranged between 3.2 and 641 ng/g lw; ∑MeO-PBDEs were between 34 and 1966 ng/g lw. Bottlenose dolphins were the most contaminated species and some individuals could present health risk for endocrine disruption since levels found were above the reported threshold (1500 ng/g lw). Stable isotope analysis was used to evaluate the biomagnification capacity of these compounds. PBDEs, MeO-PBDEs and Dec 602 showed a significant positive correlation with trophic position.

Blubber samples of Indo-Pacific bottlenose (Tursiops aduncus) and spinner (Stenella longirostris) dolphins from Zanzibar, East Africa, were analyzed for a wide range of organohalogen compounds. Methoxylated polybrominated diphenyl ethers (MeO-BDEs), presumably biogenic, were found at higher concentrations than anthropogenic organochlorine pesticides (OCPs). Only traces of industrial pollutants, such as polychlorinated biphenyls, were detected. The OCP levels found off Zanzibar were lower than those reported from other regions while MeO-BDE levels were higher. The relative composition of the OCPs indicated recent use of lindane (γ-hexachlorocyclohexane) and aged residues of DDT and technical HCH. Placental transfer was estimated to 2.5% and 0.5% of the total burden of OCPs and MeO-BDEs, respectively. Overall transfer from mother to calf in Indo-Pacific bottlenose dolphins was estimated to 72% and 85% for the OCPs and MeO-BDEs burdens, respectively. Health effects of MeO-BDEs are not known, but structural similarities with well-known environmental toxins are cause for concern.

Perfluoroalkyl substances (PFASs) are a class of organohalogenated compounds of environmental concern due to similar characteristics as the well-studied legacy persistent organic pollutants (POPs) that typically show environmental persistence, biomagnification and toxicity. Nevertheless, PFAS are still poorly regulated internationally and in many aspects poorly understood. Here, we studied liver and muscle concentrations in five cetacean species stranded at the southeastern coast of Spain during 2009–2018. Twelve of the fifteen targeted compounds were detected in >50% of the liver samples. Hepatic concentrations were significantly higher than those in muscle reflecting the particular toxicokinetics of these compounds. Bottlenose dolphins Tursiops truncatus showed the highest hepatic ΣPFAS (n = 5; 796.8 ± 709.0 ng g⁻¹ ww) concentrations, followed by striped dolphin Stenella coeruleoalba (n = 29; 259.5 ± 136.2 ng g⁻¹ ww), sperm whale Physeter macrocephalus (n = 1; 252.8 ng g⁻¹ ww), short-beaked common dolphin Delphinus delphis (n = 2; 240.3 ± 218.6 ng g⁻¹ ww) and Risso’s dolphin Grampus griseus (n = 1; 78.7 ng g⁻¹ ww). These interspecies differences could be partially explained by habitat preferences, although they could generally not be related to trophic position or food chain proxied by stable N (δ¹⁵N) and C (δ¹³C) isotope values, respectively. PFAS profiles in all species showed a similar pattern of concentration prevalence in the order PFOS>PFOSA>PFNA≈PFFUnA>PFDA. The higher number of samples available for striped dolphin allowed for evaluating their PFAS burden and profile in relation to the stranding year, stable isotope values, and biological variables including sex and length. However, we could only find links between δ¹⁵N and PFAS burdens in muscle tissue, and between stranding year and PFAS profile composition. Despite reductions in the manufacturing industry, these compounds still appear in high concentrations compared to more than two decades ago in the Mediterranean Sea and PFOS remains the dominating compound.

The bottlenose dolphin (Tursiops truncatus) is an upper trophic level predator and the most common cetacean species found in nearshore waters of southern Florida, including the Lower Florida Keys (LFK) and the Florida Coastal Everglades (FCE). The objective of this study was to assess contamination levels of total mercury (T-Hg) in skin and persistent organic pollutants (PCBs, PBDEs, DDXs, HCHs, HCB, Σ PCDD/Fs and Σ DL-PCBs) in blubber samples of bottlenose dolphins from LFK (n = 27) and FCE (n = 24). PCBs were the major class of compounds found in bottlenose dolphin blubber and were higher in individuals from LFK (Σ 6 PCBs LFK males: 13,421 ± 7730 ng g⁻¹ lipids, Σ 6 PCBs LFK females: 9683 ± 19,007 ng g⁻¹ lipids) than from FCE (Σ 6 PCBs FCE males: 5638 ng g⁻¹ ± 3627 lipids, Σ 6 PCBs FCE females: 1427 ± 908 ng g⁻¹ lipids). These levels were lower than previously published data from the southeastern USA. The Σ DL-PCBs were the most prevalent pollutants of dioxin and dioxin like compounds (Σ DL-PCBs LFK: 739 ng g⁻¹ lipids, Σ DL-PCBs FCE: 183 ng g⁻¹ lipids) since PCDD/F concentrations were low for both locations (mean 0.1 ng g⁻¹ lipids for LFK and FCE dolphins). The toxicity equivalences of PCDD/Fs and DL-PCBs expressed as TEQ in LFK and FCE dolphins is mainly expressed by DL-PCBs (81% LFK - 65% FCE). T-Hg concentrations in skin were significantly higher in FCE (FCE median 9314 ng g⁻¹ dw) compared to LFK dolphins (LFK median 2941 ng g⁻¹ dw). These concentrations are the highest recorded in bottlenose dolphins in the southeastern USA, and may be explained, at least partially, by the biogeochemistry of the Everglades and mangrove sedimentary habitats that create favourable conditions for the retention of mercury and make it available at high concentrations for aquatic predators.

A total of 164 blood samples from 16 clinically healthy bottlenose dolphins (Tursiops truncatus), were obtained from an aquarium in Spain between 2019 and 2020, as part of their preventive medicine protocol. In addition to conventional haematological and biochemical analyses, plasmatic B-esterase activities were characterised to determine the potential application of such analyses in wild counterparts. The hydrolysis rates for the substrates of acetylcholinesterase (AChE), butyrylcholinesterase (BuChE) and carboxylesterase (CE) activity in plasma were measured, the last using two commercial substrates, p-nitrophenyl acetate (pNPA) and p-nitrophenyl butyrate (pNPB). Activity rates (mean ± SEM in nmol/min/mL plasma) were (in descending order): AChE (125.6 ± 3.8), pNPB-CE (65.0 ± 2.2), pNPA-CE (49.7 ± 1.1) and BuChE (12.8 ± 1.3). These values for dolphins are reported in here for the first time in this species. Additionally, the in vitro sensitivity of two B-esterases (AChE and pNPB-CE) to chemicals of environmental concern was determined, and the protective role of plasmatic albumin assessed. Out of the B-esterases measured in plasma of dolphin, AChE activity was more responsive in vitro to pesticides, while CEs had a low response to plastic additives, likely due to the protective presence of albumin. However, the clear in vitro interaction of these environmental chemicals with purified AChE from electric eels and recombinant human hCEs (hCE1 and hCE2) and albumin, predicts their impact in other tissues that require in vivo validation. A relationship between esterase-like activities and health parameters in terrestrial mammals has already been established. Thus, B-esterase measures could be easily included in marine mammal health assessment protocols for dolphins as well, once the relationship between these measures and the animal's fitness has been established.

The potential impact of exposure to polychlorinated biphenyls (PCBs) on the health and survival of cetaceans continues to be an issue for conservation and management, yet few quantitative approaches for estimating population level effects have been developed. An individual based model (IBM) for assessing effects on both calf survival and immunity was developed and tested. Three case study species (bottlenose dolphin, humpback whale and killer whale) in four populations were taken as examples and the impact of varying levels of PCB uptake on achievable population growth was assessed. The unique aspect of the model is its ability to evaluate likely effects of immunosuppression in addition to calf survival, enabling consequences of PCB exposure on immune function on all age-classes to be explored. By incorporating quantitative tissue concentration-response functions from laboratory animal model species into an IBM framework, population trajectories were generated. Model outputs included estimated concentrations of PCBs in the blubber of females by age, which were then compared to published empirical data. Achievable population growth rates were more affected by the inclusion of effects of PCBs on immunity than on calf survival, but the magnitude depended on the virulence of any subsequent encounter with a pathogen and the proportion of the population exposed. Since the starting population parameters were from historic studies, which may already be impacted by PCBs, the results should be interpreted on a relative rather than an absolute basis. The framework will assist in providing quantitative risk assessments for populations of concern.

The presence of triclosan, a widely-used antibacterial chemical, is currently unknown in higher trophic-level species such as marine mammals. Blood plasma collected from wild bottlenose dolphins (Tursiops truncatus) in Charleston, SC (CHS) (n = 13) and Indian River Lagoon, FL (IRL) (n = 13) in 2005 was analyzed for triclosan. Plasma concentrations in CHS dolphins ranged from 0.12 to 0.27 ng/g wet weight (mean 0.18 ng/g), with 31% of the sampled individuals having detectable triclosan. The mean IRL dolphin plasma concentrations were 0.072 ng/g wet weight (range 0.025-0.11 ng/g); 23% of the samples having detectable triclosan. In the CHS area, triclosan effluent values from two WWTP were both 190 ng/L and primary influents were 2800 ng/L and 3400 ng/L. Triclosan values in CHS estuarine surface water samples averaged 7.5 ng/L (n = 18) ranging from 4.9 to 14 ng/L. This is the first study to report bioaccumulation of anthropogenic triclosan in a marine mammal highlighting the need for further monitoring and assessment. Triclosan in bottlenose dolphin plasma and their environment.