MORBILLIVIRUS INFECTION IN TWO SPECIES OF PILOT WHALE (GLOBICEPHALA SP.) FROM THE WESTERN ATLANTIC

We report evidence of enzootic morbillivirus infection among long-finned, Globicephala melas, and short-finned, G. macrorhynchus, pilot whales in the western Atlantic. A retrospective serologic survey, using five morbilliviruses, was carried out on 99 G. melas from 14 stranding events between 1982 and 1993 and from 25 G. macrorhynchus stranded in 5 events between 1986 and 1994. A blood sample was also obtained from an adult G. melas by-caught in the western North Atlantic. Tissues were collected from 24 G. melas and 15 G. macrorhynchus for histology and immunoperoxidase staining. Neutralizing antibody titers were found in 92 (92%) of 100 G. melas and 16 (64%) of 25 G. macrorynchus, and titers were highest against cetacean morbilliviruses. Seroprevalence was similar between age classes and sexes. The earliest evidence of infection was in a G. melas that stranded in 1982. Stable antibody titers were observed in pilot whales under rehabilitation for up to eight months. Clinical disease consistent with morbillivirus pneumonia was detected in a G. melas calf. Immunoperoxidase staining confirmed that viral antigen was present in the lesions. We propose that enzootic infection in pilot whales is facilitated by population size, social structure, and migration patterns. Furthermore, through mixing with other odontocetes, pilot whales could act as vectors through the Atlantic. Clinical morbillivirus infection may precipitate mass strandings of highly social odontocetes.     

Pilot Whales Attracted to Killer Whale Sounds: Acoustically-Mediated Interspecific Interactions in Cetaceans

In cetaceans’ communities, interactions between individuals of different species are often observed in the wild. Yet, due to methodological and technical challenges very little is known about the mediation of these interactions and their effect on cetaceans’ behavior. Killer whales (Orcinus orca) are a highly vocal species and can be both food competitors and potential predators of many other cetaceans. Thus, the interception of their vocalizations by unintended cetacean receivers may be particularly important in mediating interspecific interactions. To address this hypothesis, we conducted playbacks of killer whale vocalizations recorded during herring-feeding activity to free-ranging long-finned pilot whales (Globicephala melas). Using a multi-sensor tag, we were able to track the whales and to monitor changes of their movements and social behavior in response to the playbacks. We demonstrated that the playback of killer whale sounds to pilot whales induced a clear increase in group size and a strong attraction of the animals towards the sound source. These findings provide the first experimental evidence that the interception of heterospecific vocalizations can mediate interactions between different cetacean species in previously unrecognized ways.     

A simple photograph‐based approach for discriminating between free‐ranging long‐finned (Globicephala melas) and short‐finned (G. macrorhynchus) pilot whales off the east coast of the United States

Line transect based abundance estimation is complicated for long‐finned (LFPW, Globicephala melas) and short‐finned (SFPW, G. macrorhynchus) pilot whales because of their similarity in appearance and their overlapping summertime range in some areas. We developed a photograph‐based approach to distinguish between species of free‐ranging pilot whales in the northwest Atlantic. We collected skin samples and photographs during the summers of 2004–2007 and used skin samples to distinguish species based on mitochondrial DNA. Relative morphometric measurements from photographs were examined using mixed‐effect models and logistic regression. The best model among 94 candidate models had an overall classification error rate of 2.5%. We tested the presence/absence of pigmentation in four regions of the dorsal body (melon, eye, cape, and saddle) for differences. Pigmentation was present in all four regions in 100% of the SFPWs sampled. Melon patch, blaze, and saddle patch pigmentation were present in 6%, 68%, and 50%, respectively, of the LFPWs, but the cape was completely absent. Both types of analyses provided positive species discrimination of free‐ranging animals. We created a cost‐effective, simple tool which could ultimately assist in providing appropriate management, mitigation, and conservation strategies for both northwest Atlantic species of pilot whales.     

The use of DNA barcoding to monitor the marine mammal biodiversity along the French Atlantic coast

In the last ten years, 14 species of cetaceans and five species of pinnipeds stranded along the Atlantic coast of Brittany in the North West of France. All species included, an average of 150 animals strand each year in this area. Based on reports from the stranding network operating along this coast, the most common stranding events comprise six cetacean species (Delphinus delphis, Tursiops truncatus, Stenella coeruleoalba, Globicephala melas, Grampus griseus, Phocoena phocoena)and one pinniped species (Halichoerus grypus). Rare stranding events include deep-diving or exotic species, such as arctic seals. In this study, our aim was to determine the potential contribution of DNA barcoding to the monitoring of marine mammal biodiversity as performed by the stranding network.We sequenced more than 500 bp of the 5’ end of the mitochondrial COI gene of 89 animals of 15 different species (12 cetaceans, and three pinnipeds). Except for members of the Delphininae, all species were unambiguously discriminated on the basis of their COI sequences. We then applied DNA barcoding to identify some “undetermined” samples. With again the exception of the Delphininae, this was successful using the BOLD identification engine. For samples of the Delphininae, we sequenced a portion of the mitochondrial control region (MCR), and using a non-metric multidimentional scaling plot and posterior probability calculations we were able to determine putatively each species. We then showed, in the case of the harbour porpoise, that COI polymorphisms, although being lower than MCR ones, could also be used to assess intraspecific variability. All these results show that the use of DNA barcoding in conjunction with a stranding network could clearly increase the accuracy of the monitoring of marine mammal biodiversity.     

Eight polymorphic microsatellite loci for bottlenose dolphin and other cetacean species

We developed eight polymorphic microsatellite loci that can be used to elucidate population structure and aid management of bottlenose dolphins, Tursiops truncatus, in the northwest Atlantic. These loci averaged nine alleles, with no evidence for null alleles. Their cross‐species utility was examined in Cephalorhynchus commersonii, Delphinus delphis, Delphinapterus leucas, Eschrictius robustus, Globicephala macrorhynchus, Lagenorhynchus obliquidens, Orcinus orca, Steno bredanensis and Stenella clymene. On average, 75% of the loci were polymorphic in these species (range = 50–88%). Our results indicate that these loci will be useful for elucidating population structure of bottlenose dolphin as well as of other cetacean species.     

Worldwide mitochondrial DNA diversity and phylogeography of pilot whales (Globicephala spp.)

Pilot whales (Globicephala spp.) provide an interesting example of recently diverged oceanic species with a complex evolutionary history. The two species have wide but largely non‐overlapping ranges. Globicephala melas (long‐finned pilot whale; LFPW) has an antitropical distribution and is found in the cold‐temperate waters of the North Atlantic and Southern Hemisphere, whereas Globicephala macrorhynchus (short‐finned pilot whale; SFPW) has a circumglobal distribution and is found mainly in the tropics and subtropics. To investigate pilot whale evolution and biogeography, we analysed worldwide population structure using mitochondrial DNA (mtDNA) control region sequences (up to 620 bp) from a variety of sources (LFPW = 643; SFPW = 150), including strandings in New Zealand and Tasmania, and whale‐meat products purchased on the markets of Japan and Korea. Phylogenetic reconstructions failed to support a reciprocal monophyly of the two species, despite six diagnostic substitutions, possibly because of incomplete lineage sorting or inadequate phylogenetic information. Both species had low haplotype and nucleotide diversity compared to other abundant widespread cetaceans (LFPW, π = 0.35%; SFPW, π = 0.87%) but showed strong mtDNA differentiation between oceanic basins. Strong levels of structuring were also found at the regional level. In LFPW, phylogeographic patterns were suggestive either of a recent demographic expansion or selective sweep acting on the mtDNA. For SFPW, the waters around Japan appear to represent a centre of diversity, with two genetically‐distinct forms, as well as a third population of unknown origin. The presence of multiple unique haplotypes among SFPW from South Japan, together with previously documented morphological and ecological differences, suggests that the southern form represents a distinct subspecies and/or evolutionary significant unit. © 2009 The Linnean Society of London, Biological Journal of the Linnean Society, 2009, 98 , 729–744.     

Host Adaptation and Speciation through Hybridization and Polyploidy in Phytophthora

It is becoming increasingly evident that interspecific hybridization is a common event in phytophthora evolution. Yet, the fundamental processes underlying interspecific hybridization and the consequences for its ecological fitness and distribution are not well understood. We studied hybridization events in phytophthora clade 8b. This is a cold-tolerant group of plant pathogenic oomycetes in which six host-specific species have been described that mostly attack winter-grown vegetables. Hybrid characterization was done by sequencing and cloning of two nuclear (ITS and Ypt1) and two mitochondrial loci (Cox1 and Nadh1) combined with DNA content estimation using flow cytometry. Three different mtDNA haplotypes were recovered among the presumed hybrid isolates, dividing the hybrids into three types, with different parental species involved. In the nuclear genes, additivity, i.e. the presence of two alleles coming from different parents, was detected. Hybrid isolates showed large variations in DNA content, which was positively correlated with the additivity in nuclear loci, indicating allopolyploid hybridization followed by a process of diploidization. Moreover, indications of homeologous recombination were found in the hybrids by cloning ITS products. The hybrid isolates have been isolated from a range of hosts that have not been reported previously for clade 8b species, indicating that they have novel pathogenic potential. Next to this, DNA content measurements of the non-hybrid clade 8b species suggest that polyploidy is a common feature of this clade. We hypothesize that interspecific hybridization and polyploidy are two linked phenomena in phytophthora, and that these processes might play an important and ongoing role in the evolution of this genus.     

Short‐finned pilot whales (Globicephala macrorhynchus) of the Mariana Archipelago: Individual affiliations,movements, and spatial use

Little is known about short‐finned pilot whales (Globicephala macrorhynchus) in the western North Pacific outside of Japanese coastal waters. To expand understanding of short‐finned pilot whale ecology in the region, we conducted small‐boat surveys in 2010?2016 within the Mariana Archipelago to investigate individual associations, movements, spatial use, and dive behavior of short‐finned pilot whales. We collected genetic, photo‐identification, and satellite‐tag data and identified 191 distinctive individuals. A preliminary social network diagram of photo‐cataloged individuals revealed a main cluster that comprised 82% of individuals, representing all five mitochondrial DNA haplotypes identified within the population. Kernel density estimates for tagged short‐finned pilot whales (n = 11) during summer were used to identify areas with the highest probability of use (10% probability density contour), core area (50%) and home range (95%). The area with highest probability of use by short‐finned pilot whales was off the northwest side of Guam. Satellite tag data also suggest that some individuals are island‐associated year‐round. Data from five location‐dive tags demonstrated that the short‐finned pilot whales dove more often to intermediate depths at twilight and night, suggesting they may target prey that forage on the deep scattering layer as it migrates to and from the surface.     

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

An 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.     

Diphyllobothriids (Cestoda: Pseudophyllidea) from the long-finned pilot whale Globicephala melas (Traill, 1809) off the Faroe Islands,with comments on the taxonomy of Diphyllobothrium Cobbold, 1858 species from cetaceans

The taxonomy of marine species of the genus Diphyllobothrium, particularly those parasitic in cetaceans, is rather confused. During parasitological investigations of long-finned pilot whales Globicephala melas from waters off the Faroe Islands, five diphyllobothriid species were detected: Diphyllobothrium sp. (possibly D. polyrugosum), D. stemmacephalum, Diphyllobothrium sp. A, Diphyllobothrium sp. B and Diphyllobothriidae sp. D. stemmacephalum is reported for the first time from G. melas. The stituation regarding the taxonomy of Diphylobothrium species from cetaceans is briefly reviewed. It is concluded that the recent development of genetic techniques may be of great value in relation to clarifying the taxonomy of this group.     

First mitochondrial DNA analysis of the spectacled porpoise (Phocoena dioptrica) from Tierra del Fuego,Argentina

The spectacled porpoise (Phocoena dioptrica, Lahille, 1912) is one of the cetacean species about which least is known. Most information on the biology and ecology of this species has been obtained from stranded specimens and sightings at sea. In this study, analysis of 380 bp fragment of mitochondrial DNA (mtDNA) control region sequences (N = 50) was performed to provide a preliminary assessment of the genetic variation in spectacled porpoise specimens found stranded or by caught on the coast of Tierra del Fuego, Argentina. Results showed high levels of mtDNA diversity, as expected in large size and stable populations, and similar to other species of porpoises. The star-like shape phylogeny of haplotypes indicates a recent population expansion. This is the first report on the genetic variation of this species. Other lines of evidence (microsatellite loci, single-nucleotide polymorphism (SNPs)) are needed to improve our knowledge on the molecular biology of the spectacled porpoise.     

The density of odontocete integument depends on blubber lipid composition and temperature

Cetacean integument serves many functional roles, including contribution to whole body buoyancy. The blubber of the integument of different cetacean species contains varying concentrations of triacylglycerols (TAG) and wax esters (WE); generally, these lipid classes have different densities. Integument can also experience a wide range of temperatures during a dive, so its density may change with depth. The goals of this study were to measure integument density and isolated blubber lipid density in three deep‐diving odontocete species (n = 3–4)—short‐finned pilot whales (Globicephala macrorhynchus), pygmy sperm whales (Kogia breviceps), and Gervais' beaked whales (Mesoplodon europeaus)—at different temperatures (6°C–35°C), and to relate these densities to lipid content and composition. Kogia and Mesoplodon integument and isolated lipids had high WE content (78.7–99.5 wt%) and were less dense (by 1.7%–9.3%) than those of Globicephala, which were composed predominately of TAG. Generally, densities increased as temperature decreased. Changes in integument densities mirrored those of isolated lipid densities, suggesting that blubber lipids are largely responsible for the buoyant properties of cetacean integument. These data demonstrate that the contribution of the integument to whole body density depends on lipid class and temperature, and therefore may provide useful, species‐specific correction factors for diving energetics models.     

Molecular signatures of divergence and selection in closely related pine taxa

Efforts to detect loci under selection in plants have mostly focussed on single species. However, assuming that intraspecific divergence may lead to speciation, comparisons of genetic variation within and among recently diverged taxa can help to locate such genes. In this study, coalescent and outlier detection methods were used to assess nucleotide polymorphism and divergence at 79 nuclear gene fragments (1212 SNPs) in 16 populations (153 individuals) of the closely related, but phenotypically and ecologically distinct, pine taxa Pinus mugo, P. uliginosa and P. uncinata across their European distributions. Simultaneously, mitochondrial DNA markers, which are maternally inherited in pines and distributed by seeds at short geographic distance, were used to assess genetic relationships of the focal populations and taxa. The majority of nuclear loci showed homogenous patterns of variation between the taxa due to a high number of shared SNPs and haplotypes, similar levels of polymorphism, and low net divergence. However, against this common genetic background and an overall low population structure within taxa at mitochondrial markers, we identified several genes showing signatures of selection, accompanied by significant intra- and interspecific divergence. Our results indicate that loci involved in species divergence may be involved in intraspecific local adaptation.     

DNA barcoding of coastal ichthyofauna from Bahia,northeastern Brazil,South Atlantic: High efficiency for systematics and identification of cryptic diversity

In spite of the utility of DNA barcoding in discriminating marine fish species, few studies using this technique are available along the Brazilian coast, which encompasses nearly 9000 km, distinct biogeographic regions and endemism rates. Therefore, in the present work, we provided the first DNA barcode library based on sequencing of the mitochondrial gene cytochrome c oxidase subunit I (COI) for Actinopterygii fish from northeastern coast of Brazil. The database included 459 specimens from 54 genera, 31 families and 13 orders, with mean interspecific genetic distance of and mean intraspecific distance of 0.15%. As a result, the neighbor-joining (NJ) tree resulted in the molecular identification of 78 taxa. Cryptic species were indicated in Eucinostomus (mojarras) since the 82 specimens morphologically identified as Eucinostomus melanopterus or Eucinostomus sp. formed five clusters with intraspecific genetic distances up to 0.31% and lowest interspecific distance of 11.14%. Eucinostomus jonesii and Eucinostomus harengulus were identified for the first time in ichthyofauna inventories from northeastern Brazil. The present work has also added new records to the Barcode of Life Data System (BOLD), thereby favoring the identification of other specimens at different life stages or in forensic studies. The present dataset reinforces the reliability of DNA barcoding to estimate the overlooked richness of marine fish in Brazilian coast with application in conservation, ecology, systematics and fisheries.     

Unique and isolated: population structure has implications for management of the endangered New Zealand sea lion

Female otariids (eared seals) frequently display strong levels of philopatry, a behaviour that has the potential to influence population structure, particularly at the mitochondrial level. Conversely, male otariids often move between breeding colonies, likely facilitating nuclear gene flow between colonies. Such gender-specific movements have the potential to influence species population structure. Here we investigate the genetic population structure of the endangered New Zealand (NZ) sea lion, using nuclear (microsatellite) and mitochondrial molecular markers, with the intention to better inform conservation through identification of management units for the species. The strong levels of female philopatry in this species have potential to lead to population structure at the mitochondrial loci. In contrast, weak or no population structure is expected across nuclear loci. NZ sea lions were sampled from the main breeding areas across the species’ current distribution (three Auckland Islands sites, two Campbell Island sites, one Stewart Island site and one Otago Peninsula site). Individuals were screened for microsatellite (n?=?271; 16 loci) and mitochondrial (n?=?56; 1027 bp D-loop and 1189 bp cytb). Despite a small (c. 9880 individuals) population size, moderate levels of microsatellite variation are observed in the NZ sea lions, in contrast to low levels of mitochondrial genetic variation. Results from mitochondrial DNA analyses revealed no population structure, suggesting that the strong level of female philopatry in NZ sea lions alone is not sufficient to maintain genetic population structure. Due to the frequent male movements between breeding colonies, no population structure was detected across the nuclear loci either. The absence of genetic structure suggests that, from a genetic perspective, NZ sea lions can be considered to be a single population. Despite this, the differing impacts of threats (e.g. fisheries by-catch) to each individual breeding colony must also be taken into consideration when defining management units for this endangered species.     

Genetic study of pike perch Sander lucioperca (Linnaeus, 1758) and bersh Sander volgensis (Gmelin, 1789) from the fishery water-bodies of Kazakhstan

The genetic polymorphism of the pike perch Sander lucioperca, sampled on the territory of the Republic of Kazakhstan (Lake Balkhash, Urals River, Syrdarya River and Shardara water reservoir, North Aral Sea) and in the Volga River, and the bersh Sandler volgensis (Lake Balkhash, Volga River) populations was examined at six microsatellite loci and the mitochondrial DNA cytochrome b gene. All examined loci were successfully amplified in both species and produced quite different allele profiles, which allows them to be used for accurate species identification of pike perch and bersh. At the same time, neither nuclear nor mitochondrial markers revealed statistically significant differentiation between the examined pike perch samples, excluding some isolation of the population from Volga River.     

Inter-species differences in polychlorinated biphenyls patterns from five sympatric species of odontocetes: Can PCBs be used as tracers of feeding ecology?

Concentrations of thirty two polychlorinated biphenyls (PCBs) were determined in the blubber of five sympatric species of odontocetes stranded or by-caught along the Northwest coast of the Iberian Peninsula: common dolphin (Delphinus delphis), long-finned pilot whale (Globicephala melas), harbour porpoise (Phocoena phocoena), striped dolphin (Stenella coeruleoalba) and bottlenose dolphin (Tursiops truncatus). Multivariate analyses were applied to evaluate the ability of PCB patterns to discriminate these sympatric species and to determine which eco-biological factors influence these patterns, thus evaluating the relevance of PCB concentrations as biogeochemical tracers of feeding ecology. The five species could be separated according to their PCB patterns. Different exposure to these contaminants, a consequence of their different dietary preferences or habitats, together with potentially dissimilar metabolic capacities, likely explain these results; sex, age, habitat and the type of prey eaten were the most important eco-biological parameters of those tested. Although, no single congener has been specifically identified as a tracer of feeding ecology, 4 congeners from the 22 analysed seemed to be the most useful and around 12 congeners appear to be enough to achieve good discrimination of the cetaceans studied. Therefore, this study suggests that PCB patterns can be used as tracers for studying the feeding ecology, sources of contamination or even population structure of cetacean species from the Northwest Iberian Peninsula.     

Stomach contents of long-finned pilot whales (Globicephala melas) from southern Chile

The widely distributed long-finned pilot whale (Globicephala melas) has been reported off the Chilean coast, from Iquique (20°12′S) south to Navarino Island (55°15′S; 67°30′W), but little is known about its biology or ecology in the region. Here, we report on the prey of this species, identified by stomach content analyses from animals stranded on Holget Islets, Beagle Channel, southern Chile in August 2006. The stomachs of seven individuals (six females and one male) contained cephalopod remains. The prey composition found in these southern Chilean pilot whales was similar to that described in other parts of the world and the Southern Ocean. This is the first report on the feeding habits of this species from Chile.