Feresa Research Articles

Toxicogenomic assessment of hydroxylated metabolites of PBDEs on cetaceans: An in vitro study

Despite their ban, polybrominated diphenyl ethers (PBDEs) are frequently detected in various environmental compartments including marine and coastal ecosystems due to their persistence, bio-accumulative, high production volumes, and widespread use. One of the major concerns from PBDEs is the transformation products, such as hydroxylated polybrominated diphenyl ethers (OH-BDEs), which are more bioactive than the parent compounds. For example, 6-hydroxy-2,2′,4’,4-tetrabromodiphenyl ether (6-OH-BDE-47) is a typical metabolite of PBDEs and cause endocrine system disruption, developmental toxicity, and neurotoxicity in different species. Despite being widely detected in marine environments, investigations on the toxicological mechanisms of 6-OH-BDE-47 in cetaceans remain scarce. High concentrations of PBDEs accumulate in cetaceans due to the long lifespan and large fat reserve. The accumulated PBDEs have become the major source of OH-BDEs in cetaceans. We exposed immortalized fibroblast cell lines from the skin of pygmy killer whales (PKW-LWHT) and Indo-Pacific finless porpoises (FP-LWHT) to 6-OH-BDE-47 and analyzed changes in cellular function using transcriptomic data, along with enzymatic activity. Exposure to the body-relevant body burdens of 6-OH-BDE-47 (250 and 500 ng mL−1) significantly decreased cell viability. Differentially expressed genes in FP-LWHT exposed to 6-OH-BDE-47 were primarily enriched in the pathways associated with steroid metabolism. Total cholesterol was decreased by 6-OH-BDE-47, whereas low-density lipoprotein cholesterol and triglyceride levels were significantly increased in FP-LWHT cells. In contrast, glycolysis was the main enriched function of differentially expressed genes in PKW-LWHT cells exposed to 6-OH-BDE-47, and the enzyme activity of phosphofructokinase and hexokinase was upregulated. Thus, even though the cell viability of both cell lines from these two species was significantly suppressed by 6-OH-BDE-47, the cellular response or affected cellular function was different between the Pygmy killer whale and the Indo-Pacific Finless Porpoise, suggesting a diverse response towards OH-BDEs exposure.

Cetacean Stranding Response Program and Spatial-Temporal Analysis in Taiwan, 1994-2018.

A national cetacean stranding response program in Taiwan has evolved significantly in the past three decades. Initially co-ordinated by National Taiwan University from 1994, the program transitioned to the Taiwan Cetacean Society in 1999, and local governments took on a more prominent role after 2009. A comprehensive stranding database (1994-2018) has been maintained, which documented 1320 stranding events involving 1698 animals from at least 27 species. The most commonly stranded species include finless porpoises, bottlenose dolphins, Kogia spp., and Risso's dolphins. The stranding rates varied annually and seasonally, with increases noted from an average of 16 events per year for the first 3 years to 44-58 events per year between 1997 and 2015, and a sharp rise to over 90 events per year for the period of the last three years. Seasonal variations were also significant, with higher stranding rates during the northeastern monsoon (NEM, October to next April) than that during southwestern monsoon (SWM, May to September). From the aspect of distribution, more frequent and even strandings occurred along the coast of northern Taiwan, while mass strandings were concentrated in the southwestern counties during NEM. Among all strandings, 390 events (29.5%) and 660 animals (38.9%) were live ones. Under great effort in rescuing and rehabilitating 52 cases, 15 cetacean individuals have been released since 2000. Additionally, there have been 56 mass strandings involving at least 11 species since 1994, predominated by pygmy killer whales, particularly during the NEM season along the southwest coast. This study not only contributes to our understanding of the stranding patterns and diversity of the cetaceans in Taiwan, but also provides valuable insights for future conservation strategies on cetaceans in the western Pacific.

Assessing animal welfare during a stranding of pygmy killer whales (Feresa attenuata)

AbstractEmpirical assessment of cetacean welfare to inform stranding interventions is lacking. Here, potential welfare indicators are described for two stranded pygmy killer whales (Feresa attenuata), along with euthanasia procedures and pathology of potential relevance. The animals were filmed for 3.5 and 1.5 hr, respectively, allowing assessment of 19 indicators, including animal behaviors and human interventions. Eight interventions and 19 animal behaviors were identified; 17 and 11 behaviors were displayed by animal 1 and 2, respectively. Examination of ballistics euthanasia revealed atypical projectile placement and characterized animal behavioral responses, but welfare implications could not be assessed as insensibility was not verified in‐field. Pulmonary edema and renal degeneration were documented in both animals; differential etiologies include ischemia–reperfusion, shock, and/or myopathy. Potential relationships among histopathology and welfare indicators are explored to infer affective experiences. For example, simultaneous head‐lifting with respiration increased over time which, alongside pulmonary edema, suggests these animals experienced breathlessness. Other likely affective states include fatigue and discomfort; there are insufficient data to estimate the intensity or duration of these experiences or to provide an overall welfare grade/score. Further data are required to validate the proposed welfare indicators and to progress development of holistic approaches to welfare assessment at cetacean strandings.

Observations of cetaceans in the Maldives, 1990-2002

Cetaceans observed in Maldivian waters were recorded during the period August 1990 to June 2002, from both vessels-of-opportunity and dedicated cetacean-watching cruises. A total of 1,829 cetacean sightings were recorded during 535 days at sea (equivalent to 261 standardised days). There were 83 multispecies sightings, plus a further 58 sightings without associated effort data and 129 strandings were recorded by the author and others, making a total of 2,108 cetacean records. In all, 20 different species were positively identified from sightings. Spinner dolphins were the most abundant species seen, accounting for 35% of sightings and 53% of numbers. This species showed a clear diurnal pattern of behaviour, with many schools entering the atolls in the early morning, and leaving in the late afternoon. Spinner dolphins regularly occurred with pantropical spotted dolphins and both species associated with yellowfin tuna. Bryde’s whales also associated with yellowfin tuna and appeared to be most common in Maldivian waters during El Niño Southern Oscillation events. Blue whales were only recorded during November to April. Dwarf sperm whales were especially difficult to locate in rough weather but relatively common, making up one sixth of all sightings in flat-calm conditions. Melon-headed whales were particularly common in the south of the Maldives, but rare in the centre and north. Other species recorded were humpback whale, sperm whale, rough-toothed dolphin, Risso’s dolphin, bottlenose dolphin, striped dolphin, Fraser’s dolphin, pygmy killer whale, false killer whale, killer whale, short-finned pilot whale, Blainville’s beaked whale, Longman’s beaked whale and Cuvier’s beaked whale.

Unusual cetacean stranding events of Taiwan in 2004 and 2005

In early 2004 and in 2005, several unusual stranding events occurred in Taiwan during a period when large-scale naval exercises were conducted in and on nearby waters. Gross examination of the partial remains of two carcasses (a ginkgo-toothed beaked whale (Mesoplodon ginkgodens) and a pygmy killer whale (Feresa attenuata)) and an intact Risso’s dolphin (Grampus griseus) revealed that the former two had internal injuries to structures associated with or related to acoustics or diving. The several unusual stranding events and the findings of the gross post mortem examination of the only specimens that were available for study were suggestive that nearby naval exercises may have contributed to or caused the death of at least one cetacean in this region and that species other than beaked whales may also be susceptible to such activities. With an increasing number of military exercises in this region, more attention to the impacts of such activities on cetaceans is needed.

Biologically Important Areas II for cetaceans within U.S. and adjacent waters – Hawaiʻi Region

In this assessment we incorporated published and unpublished information to delineate and score Biologically Important Areas (BIAs) for cetaceans in the Hawaiʻi region following standardized criteria. Twenty-six cetacean species have been documented in Hawaiʻi. Eleven odontocete species have distinct small populations resident to one or more island areas: rough-toothed dolphins, pantropical spotted dolphins, common bottlenose dolphins, spinner dolphins, short-finned pilot whales, false killer whales, pygmy killer whales, melon-headed whales, Blainville’s beaked whales, Cuvier’s beaked whales, and dwarf sperm whales. Eight species of mysticetes have been documented, although their occurrence and behavior are poorly understood, with the exception of breeding humpback whales and, more recently, common minke whales. Thirty-five BIAs were delineated or revised from the initial 2015 effort: 33 for small and resident odontocete populations and two for humpback whale reproductive areas. Hierarchical BIAs reflecting core areas of use or population-specific ranges were delineated for nine species. Reproductive watch list areas were designated for common minke whales in the main Hawaiian Islands (MHI) and humpback whales in the Northwestern Hawaiian Islands (NWHI); these areas did not meet the criteria for a BIA due to limited supporting information. All but three BIAs were in the MHI, reflecting the disparities in research effort between this region and the NWHI. Spatial extents of BIA boundaries ranged from 457 km2to 138,001 km2(median = 8,299 km2). Scores (range: 1-3) for Data Support and Boundary Certainty were moderate to high (mean = 2.40 and 2.43, respectively), while Intensity and Importance scores were slightly lower (mean = 1.94 and 1.89, respectively). Many of the Hawaiʻi species have been extensively studied over several decades; accordingly, this region ranks among the highest in terms of Data Support relative to other regions. BIAs presented here describe known ranges of small resident populations, intensities of use, and uncertainties in important areas for cetaceans in Hawaiʻi based on the best available data, and have also revealed knowledge gaps to guide future research efforts.

Audiograms and click spectra of seven novel and seldom-tested odontocetes

The use of auditory evoked potentials has been promoted as a means by which to collect audiometric information from odontocete cetaceans that are rarely encountered in stranding situations. This article presents the results of auditory evoked potential hearing tests collected from stranded odontocetes over nearly a decade. For six species, no audiograms previously existed – the dwarf sperm whale (Kogia sima), pygmy sperm whale (Kogia breviceps), northern right whale dolphin (Lissodelphis borealis), melon-headed whale (Peponocephala electra), long-beaked common dolphin (Delphinus capensis), and Atlantic spotted dolphin (Stenella frontalis). Additional hearing information was gathered for the pygmy killer whale (Feresa attenuata), a species for which only two prior audiograms had been collected. Audiograms for the delphinids demonstrated a typical dolphin-like form with upper-frequency limits of hearing > 149 kHz, except for the pygmy killer whales whose upper-frequency limit was between 103-107 kHz. The kogiid audiograms had a narrower region of increased sensitivity (80-128 kHz) closely aligned with their narrowband, high-frequency echolocation signals. Distinctions between kogiids and delphinids existed in the latencies of peaks of click-evoked auditory brainstem responses, with longer interwave intervals between P4 and N5 in the kogiids (mean of 0.60 ms vs. a mean of 0.37 ms in the delphinids). Modulation rate transfer functions collected in three of the species, suggested group-wide similarities in temporal processing capabilities.

Interspecies differences in mammalian susceptibility to legacy POPs and trace metals using skin fibroblast cells

The susceptibility to trace metals and legacy POPs is different between terrestrial and marine mammals. In this study, we established the first cell line from Indo-Pacific finless porpoises and compared the cellular responses of skin fibroblast cells from Pygmy killer whales, Pantropic spotted dolphins, Indo-Pacific finless porpoises, mice, and humans following exposure to copper, methylmercury, cadmium, PCB126, PCB153, and BDE47 to better understand the interspecies sensitivities of mammals to chemical pollutants. We conducted a risk assessment by comparing no-observed effect concentrations (NOEC), lowest-observed effect concentrations (LOEC), and half maximal effective concentrations (EC50) from cell viability assays and previously reported pollutant body burdens in mammals. Based on the in vitro data, Indo-Pacific finless porpoises were more sensitive to copper and methylmercury than other mammals. PCB153 exposure reduced cell viability in all mammals except humans, while PCB126 was more potent, with 13.33 μg/mL exposure reducing cell viability in all mammals. In contrast, BDE47 exposure reduced cell viability only in terrestrial mammals in addition to pantropic spotted dolphin. Based on the in vitro data and the natural context of metal concentrations, both methylmercury and cadmium posed a higher risk to cetaceans than human, while copper posed a lower risk to cetaceans. All three legacy POPs (PCB126, PCB153, and BDE47) posed minor risk to cetaceans for short-term exposure. This study demonstrated that a species-specific in vitro model may provide more accurate information on the potential risk of pollutants to mammals. However, due to the bioamplification of POPs and their potential impact on the endocrine system and immune system of cetaceans, risk assessment with long-term exposure with more in vitro models should be further studied.

Sexually dimorphic characteristics of short‐finned pilot whales, false killer whales, pygmy killer whales, and melon‐headed whales assessed using fin and body morphometrics from photographs taken at sea

AbstractAdults of many free‐ranging delphinid species cannot be reliably sexed at sea. Sexually mature, known‐sex adult short‐finned pilot, pygmy killer, melon‐headed, and false killer whales were profiled from at‐sea photos to assess proportional fin and body dimorphism. Males of all four species had larger dorsal fins proportional to anterior body length. False killer whales showed no further dimorphism, while melon‐headed whales showed subtle dimorphism in dorsal fin overhang. Pygmy killer whales showed subtle dimorphism in lateral positioning of the top‐most point and overhang. Lastly, short‐finned pilot whales showed strong dimorphism in the leading edge, lateral positioning of the top‐most point, and height of the overhang. Generalized linear models showed strong predictive accuracy for pilot and false killer whales, and moderate accuracy for pygmy killer and melon‐headed whales. Results indicate that adult short‐finned pilot and false killer whales can be accurately sexed in the field or via morphometrics. Adult pygmy killer and melon‐headed whales have subtle sexual dimorphism that can be detected using morphometrics. The presence of dimorphism gives insight into potential reproductive strategies and social structure. Further analysis is needed to explore morphometric differences between inshore and pelagic populations of these species, as well as ontogenetic growth between life stages.

Auditory evoked potential in stranded melon-headed whales (Peponocephala electra): With severe hearing loss and possibly caused by anthropogenic noise pollution

Highly concentrated live mass stranding events of dolphins and whales happened in the eastern coast of China between June and October 2021. The current study adopted the non-invasive auditory evoked-potential technique to investigate the hearing threshold of a stranded melon headed whale (Peponocephala electra) at a frequency range of between 9.5 and 181 kHz. It was found that, at the frequency range of from 10 to 100 kHz, hearing thresholds for the animal were between 20 and 65 dB higher than those of its phylogenetically closest species (Pygmy killer whale). The severe hearing loss in the melon headed whale was probably caused by transient intense anthropogenic sonar or chronic shipping noise exposures. The hearing loss could have been the cause for the observed temporal and spatial clustered stranding events. Therefore, there is need for noise mitigation strategies to reduce noise exposure levels for marine mammals in the coastal areas of China.

6-OH-BDE-47 inhibited proliferation of skin fibroblasts from pygmy killer whale by inducing cell cycle arrest

Hydroxylated polybrominated diphenyl ethers (OH-BDEs) are major transformation products of PBDEs that readily bioaccumulate in the marine food web. Although 6-OH-BDE-47 is frequently and abundantly detected in cetaceans, its potential toxic effects are largely unknown. We explored the toxicological pathways and mechanisms of OH-BDEs by exposing pygmy killer whale skin fibroblast cell lines (PKW-LWHT) to 6-OH-BDE-47 at concentrations ranging from 0.02, 0.2, 2 to 4 μM. The result showed that 6-OH-BDE-47 inhibited cell proliferation in a concentration- and time-dependent manner. The cell cycle data revealed that the cell cycle was arrest at the G0/G1 phase by 6-OH-BDE-47. Using qPCR and Western blot assay, we found that 6-OH-BDE-47 up-regulated the transcription and expression level of p21 and RB1 and down-regulated the expression level of Proliferating Cell Nuclear Antigen (PCNA), CDK2, CDK4, cyclin D1, cyclin E2, E2F1, and E2F3 and the cellular phosphorylated RB1. The results showed that 6-OH-BDE-47 was able to arrest the cell cycle of PKW-LWHT cells at G1 phase by changing the expression level of related regulatory genes in G1 stage, and finally inhibit cell proliferation.

Rapid weight loss in free ranging pygmy killer whales (Feresa attenuata) and the implications for anthropogenic disturbance of odontocetes

Understanding the impacts of foraging disruptions to odontocete body condition is fundamental to quantifying biological effects of human disturbance and environmental changes on cetacean populations. Here, reductions in body volume of free-ranging pygmy killer whales (Feresa attenuata) were calculated using repeated measurements of the same individuals obtained through Unoccupied Aerial System (UAS)-photogrammetry during a prolonged disruption in foraging activity arising from a 21-day stranding event. Stranded individuals were used to verify UAS-derived volume and length estimates through 3D-imaging, water displacement, and post-mortem measurements. We show that (a) UAS estimates of length were within 1.5% of actual body length and UAS volume estimates were within 10–13% of actual volume, (b) foraging disruption resulted in a daily decrease of 2% of total body mass/day, and (c) pygmy killer whales can lose up to 27% of their total body weight within 17 days. These findings highlight the use of UAS as a promising new method to remotely monitor changes in body condition and animal health, which can be used to determine the potential effects of anthropogenic disturbance and environmental change on free-ranging odontocetes.

Cetacean diversity of Mauritius

There is limited information concerning the cetaceans inhabiting the coastal waters of Mauritius. This document details the sightings of cetaceans encountered during April 2008 – September 2014. Boat-based surveys were conducted throughout the year, primarily focusing on spinner and bottlenose dolphin populations that are important for the local dolphin-watching industry. More than 1246 hours over 749 days of surveys were spent searching for cetaceans. All cetaceans encountered were recorded. During 2013 and 2014, dedicated diversity surveys were conducted and 2443.6km of search effort was covered around the islands. The encounter rate was 0.025 cetacean sightings/km. Over the total study period 13 species were recorded from 1459 sightings. Species included: Megaptera novaeangliae, Stenella longirostris, Tursiops aduncus, Tursiops truncatus, Globicephala macrorhynchus, Stenella attenuata, Physeter macrocephalus, Mesoplodon densirostris, Feresa attenuata, Peponocephala electra, Grampus griseus and Ziphius cavirostris. In addition, there were strandings of Kogia sima and Ziphius cavirostris. The most common species encountered were spinner dolphins, Indo-Pacific bottlenose dolphins, humpback whales and sperm whales. As expected only humpback whales showed any seasonality in monthly encounter rates (Kruskal-Wallis H = 42.39, Hc = 54.25, p < 0.001). The overall Shannon diversity index was 0.58 (95% CI 0.57–0.60) though this ranged between 0.77 in 2009 to 0.25 in 2014. The Shannon index value for the diversity study was 0.83 (95% CI 0.74–0.91). This study demonstrates the richness of the waters around Mauritius and the importance of the area for beaked whales, migrating humpback whales and the presence of resident sperm whales.

Maximum entropy model: Estimating the relative suitability of cetacean habitat in the northern Savu Sea, Indonesia

AbstractAn understanding of cetacean distribution is necessary to gain insights into crucial ecological processes for species conservation management. However, cetacean habitat preference and distribution in the northern Savu Sea, Indonesia, are still poorly understood. We use maximum entropy modeling with five environmental predictors to describe habitat preference and distribution of seven cetacean species in the northern Savu Sea. Our study confirms that static predictors are the most important variables in explaining habitat preferences of seven cetacean species in the northern Savu Sea. Seasonally, each cetacean species has a different environmental preference. Globicephala macrorhynchus prefers the open nearshore areas adjacent to deep waters. Stenella attenuata is associated with nearshore habitats and higher productivity areas. Stenella longirostris prefers nearshore habitat during coastal upwelling events. Tursiops truncatus prefers nearshore sheltered habitat. Grampus griseus habitats are highly related to depth and steep gradients around the isobaths of 200–1,000 m. Feresa attenuata are associated with deep waters and cooler sea temperature. Pseudorca crassidens prefers protective bays with productive waters. This study provides important information for reviewing marine spatial plans of the northern Savu Sea, specifically on managing species fisheries interaction, optimizing surveillance, and regulating marine traffic.

Marine mammals used as bait for improvised fish aggregating devices in marine waters of Ecuador, eastern tropical Pacific

Fish aggregating devices (FADs) are floating objects typically used to attract and capture pelagic fish in industrial tuna fisheries. This study documents 9 cases, involving 31 marine mammals, of incidentally captured, killed or otherwise retrieved cetaceans and pinnipeds which were used, or presumably used, as bait for improvised fish aggregation devices (IFAD) by artisanal fishers in coastal Ecuador. At least 3 species of small cetaceans were affected, including pantropical spotted dolphinStenella attenuata, short-finned pilot whaleGlobicephala macrorhynchus, pygmy killer whaleFeresa attenuataand an unidentified small delphinid, as well as South American sea lionsOtaria byroniawhich were reportedly killed on purpose for this fishing practice. A sperm whalePhyseter macrocephalusand a humpback whaleMegaptera novaeangliaewere presumably found floating at sea and opportunistically exploited as FADs. The South American sea lion represented 80.6% of marine mammals used as bait associated with FADs (25 sea lions out of 31 marine mammals), while the remaining 5 (possibly 6) cetacean species represented 19.4%. This is the first report of baited FADs in Ecuador, the extent of which is still unknown. This fishing technique has not been documented in other nations along the west coast of South America, although baiting of gillnets with marine mammal parts is common in Peru. Without fisheries management and regulation, this illegal fishing practice could rapidly expand and lead to further direct kills and conservation problems for targeted marine mammal populations in the eastern tropical Pacific. A bottom-up fisheries policy in concert with community-based conservation to ban the use of marine mammals as FAD bait is recommended.

Cetacean Distribution and Diversity in Lakshadweep Waters, India, Using a Platform of Opportunity: October 2015 to April 2016

Prior stranding records suggest that at least 12 cetacean species occur within the Lakshadweep archipelago off the southwest coast of India. These islands consist of coral atolls and form the northern part of the undersea Chagos-Laccadive ridge. Distinct oceanographic features, seasonal monsoon cycles, and high productivity make this region a potentially rich cetacean habitat In this article, we report findings from the first systematic visual cetacean surveys, which were conducted from high-speed passenger ferries that sail between nine Lakshadweep islands. The surveys were carried out between October 2015 and April 2016 during both the northeast monsoon (October to December) and inter-monsoon (January to April) seasons. We used a line-transect survey framework to record sightings as well as group size estimates. We documented 139 sightings over 3,880 km of which 78 sightings were during systematic survey effort. Eight odontocete species were confirmed from these sightings: Stenella longirostris, S. attenuata, S. coeruleoalba, Tursiops spp., Globicephala macrorhynchus, Pseudorca crassidens, Grampus griseus, and Feresa attenuata. One Balaenoptera sp. was also encountered during this survey. S. longirostris was sighted the most often (n = 22) followed by Tursiops spp. (n = 18) and G. macrorhynchus (n = 13). We documented significantly higher sightings in the northeast monsoon season compared to the intermonsoon season. Along ferry routes, cetacean species differed significantly from each other with respect to their associations with seafloor slope gradients and distances to nearest landmass. We encountered mixed species assemblages of G. macrorhynchus with Tursiops sp. and S. attenuata with Tursiops sp. Given the confirmed high cetacean diversity and occurrence in this region, there is a need for in-depth, long-term studies on biogeography, ecology, and population status of cetaceans here.

How to tell them apart? Discriminating tropical blackfish species using fin and body measurements from photographs taken at sea

AbstractThe misidentification of species and populations is a hindrance to effective cetacean management. We devised a method of species identification using 10 fin and body measurements obtainable from at‐sea photographs, and demonstrated its ability to distinguish four species of Hawaiian “blackfish”: pygmy killer, melon‐headed, short‐finned pilot, and false killer whales. Measurements from photos of 382 known individuals were converted into 14 ratios and reduced using nonmetric multidimensional scaling and principal component analysis. The first three components of the PCA explained 81% of variance among species. Ratios indicated by PCA and NMDS were tested using analysis of variance, and results identified three fin ratios that had distinct means across all four species: height/base, depth (from anterior insertion) at trailing edge apex/depth at topmost point, and width (from leading edge) at posterior point/base. Dual analysis of adults and all age classes showed near‐identical sources of variance, 90% similarity in interspecies ratio relationships, and overlapping ratio means and ranges. Results suggest similar ontogenetic growth across these four species, and confirm the efficacy of this discrimination technique for all age classes. This study established a reliable means of distinguishing these cetacean species, which will improve the efficacy of management in areas with sympatric distributions.

Survey of Brucella spp. and Leptospira spp. antibodies in cetaceans and manatees of the Amazon basin and Atlantic Ocean, Brazil

Aquatic mammals can act as sentinels of emerging and resurging pathogens in the environment. Brucella spp. and Leptospira spp. are 2 zoonotic pathogens relevant to aquatic mammals, and their detection can be used to assess pathogen exposure. In this study, serum from 84 individuals-63 cetaceans (families Iniidae, n = 37; Delphinidae, n = 22; and Kogiidae, n = 4) and 21 West Indian manatees Trichechus manatus-was tested by the Rose Bengal Test (RBT) and a commercial competitive enzyme-linked immunosorbent assay (c-ELISA) for detecting Brucella spp. antibodies, and the microscopic agglutination test (MAT) for screening Leptospira spp. exposure. Overall, 4.8% (3/63) of cetaceans were positive by RBT and 15.9% (10/63) by c-ELISA for Brucella spp. Serum from 8 c-ELISA positive cetaceans (with available serum) was further tested via serum agglutination test (SAT) and 1 individual was positive. c-ELISA was more sensitive than RBT. Exposure to Brucella spp. was found in 5 cetacean species: Clymene dolphin Stenella clymene, short-finned pilot whale Globicephala macrorhynchus, pygmy killer whale Feresa attenuata, melon-headed whale Peponocephala electra and Atlantic bottlenose dolphin Tursiops truncatus in the Atlantic Ocean, Brazil, expanding the range of known Brucella seropositive aquatic hosts. No evidence of Brucella spp. exposure was found in Iniidae and Kogiidae odontocetes and manatees. Antibodies against Leptospira spp. were not detected in cetaceans and sirenians by MAT. These results contribute to the evaluation of different Brucella spp. serological methods in cetaceans and manatees and highlight the epidemiology of zoonotic pathogens in aquatic mammals of the southwestern Atlantic Ocean and the Amazon basin.

Movements and Dive Patterns of Pygmy Killer Whales (Feresa attenuata) Released in the Gulf of Mexico Following Rehabilitation

Movements and Dive Patterns of Pygmy Killer Whales (Feresa attenuata) Released in the Gulf of Mexico Following Rehabilitation

Establishment and characterization of pygmy killer whale (Feresa attenuata) dermal fibroblast cell line.

The pygmy killer whale (Feresa attenuata) (PKW) is a tropical and subtropical marine mammal commonly found in the Atlantic, Indian and Pacific oceans. Since the PKWs live in offshore protected territories, they are rarely seen onshore. Hence, PKW are one of the most poorly understood oceanic species of odontocetes. The dermal tissue comes primarily from stranding events that occur along the coast of the Shantou, Guangdong, China. The sampled tissues were immediately processed and attached on collagen-coated 6-well tissue culture plate. The complete medium (DMEM and Ham’s F12, fetal bovine serum, antibiotic and essential amino acids) was added to the culture plates. The primary culture (PKW-LWH) cells were verified as fibroblast by vimentin and karyotype analyses, which revealed 42 autosomes and two sex chromosomes X and Y. Following transfection of PKW-LWH cells with a plasmid encoding, the SV40 large T-antigens and the transfected cells were isolated and expanded. Using RT-PCR, western blot, immunofluorescence analysis and SV40 large T-antigen stability was confirmed. The cell proliferation rate of the fibroblast cells, PKW-LWHT was faster than the primary cells PKW-LWH with the doubling time 68.9h and 14.4h, respectively. In this study, we established PKW dermal fibroblast cell line for the first time, providing a unique opportunity for in vitro studies on the effects of environmental pollutants and pathogens that could be determined in PKW and/or Cetaceans.