Corticosterone in central North Pacific male humpback whales (Megaptera novaeangliae): Pairing sighting histories with endocrine markers to assess stress.

Developing a better understanding of the stress response is critical to ensuring the health and sustainability of marine mammal populations. However, accurately measuring and interpreting a stress response in free-ranging, large cetaceans is a nascent field. Here, an enzyme immunoassay for corticosterone was validated for use in biopsy samples from male humpback whales (Megaptera novaeangliae). Analyses were conducted on 247 male North Pacific humpback whale blubber samples, including 238 non-calves and 9 calves that were collected on the Hawaiian breeding and Southeast Alaskan feeding grounds from 2004 to 2006. Significant relationships were found when corticosterone concentrations were examined by year, age class and distribution between locations. When examined by year, corticosterone concentrations for male humpback whales were higher in Hawaii in 2004 than in 2005 and 2006 (p < 0.05). Corticosterone concentration also varied by age class with initially high concentrations at birth which subsequently tapered off and remained relatively low until sexual maturity was reached around age 8-10 years. Corticosterone concentrations appeared to peak in male humpback whales around 15-25 years of age. Blubber biopsies from Alaska and Hawaii had similar mean corticosterone concentrations, yet the variability in these samples was much greater for whales located in Hawaii. It is clear that much work remains to be done in order to accurately define or monitor a stress response in male humpback whales and that specific attention is required when looking at age, sex, and yearly trends. Our results suggest that a stress response may be most impacted by age and yearly oceanographic conditions and needs to be initially examined at the individual level.

Lifetime extension of humpback whale skin fibroblasts and their response to lipopolysaccharide (LPS) and a mixture of polychlorinated biphenyls (Aroclor).

Marine mammals, such as whales, have a high proportion of body fat and so are susceptible to the accumulation, and associated detrimental health effects, of lipophilic environmental contaminants. Recently, we developed a wild-type cell line from humpback whale fibroblasts (HuWa). Extensive molecular assessments with mammalian wild-type cells are typically constrained by a finite life span, with cells eventually becoming senescent. Thus, the present work explored the possibility of preventing senescence in the HuWa cell line by transfection with plasmids encoding the simian virus large T antigen (SV40T) or telomerase reverse transcriptase (TERT). No stable expression was achieved upon SV40 transfection. Transfection with TERT, on the other hand, activated the expression of telomerase in HuWa cells. At the time of manuscript preparation, the transfected HuWa cells (HuWaTERT) have been stable for at least 59 passages post-transfection. HuWaTERT proliferate rapidly and maintain initial cell characteristics, such as morphology and chromosomal stability. The response of HuWaTERT cells to an immune stimulant (lipopolysaccharide (LPS)) and an immunotoxicant (Aroclor1254) was assessed by measurement of intracellular levels of the pro-inflammatory cytokines interleukin (IL)-6, IL-1ß and tumour necrosis factor (TNF)-α. HuWaTERT cells constitutively express IL-6, IL-1ß and TNFα. Exposure to neither LPS nor Aroclor1254 had an effect on the levels of these cytokines. Overall, this work supports the diverse applicability of HuWa cell lines in that they display reliable long-term preservation, susceptibility to exogenous gene transfer and enable the study of humpback whale-specific cellular response mechanisms.

Testosterone trends within and across seasons in male humpback whales (Megaptera novaeangliae) from Hawaii and Alaska.

Understanding reproductive profiles and timing of reproductive events is essential in the management and conservation of humpback whales (Megaptera novaeangliae). Yet compared to other parameters and life history traits, such as abundance, migratory trends, reproductive rates, behavior and communication, relatively little is known about variations in reproductive physiology, especially in males. Here, an enzyme immunoassay (EIA) for testosterone was validated for use in biopsy samples from male humpback whales. Analyses were conducted on 277 North Pacific male humpback whale blubber samples, including 268 non-calves and 9 calves that were collected in the Hawaiian breeding grounds and the Southeast Alaskan feeding grounds from 2004 to 2006. Testosterone concentrations (ng/g) were significantly different between non-calves sampled in Hawaii (n = 182) and Alaska (n = 86, p < 0.05) with peak testosterone concentrations occurring in the winter (January-March) and the lowest concentrations occurring in the summer (June-September). Fall and spring showed increasing and decreasing trends in testosterone concentrations, respectively. Blubber testosterone concentrations in non-calves and calves sampled in Alaska were not significantly different. Blubber and skin from the same individual biopsies (n = 37) were also compared, with blubber having significantly higher testosterone concentrations (p < 0.05) than skin samples. We found variability in testosterone concentration with age, suggesting that male humpbacks reach peak lifetime testosterone concentrations in the breeding grounds around age 8-25 years. The testosterone profile of male humpback whales follows a predictable pattern for capital breeders, where testosterone begins to increase prior to the breeding season, stimulating the onset of spermatogenesis. Incorporation of reproductive hormonal profiles into our overall understanding of humpback whale physiology will shed additional light on the timing of reproduction and overall health of the recently delisted Hawaii distinct population segment (DPS).

New views of humpback whale flow dynamics and oral morphology during prey engulfment.

The rise of inexpensive, user-friendly cameras and editing software promises to revolutionize data collection with minimal disturbance to marine mammals. Video sequences recorded by aerial drones and GoPro cameras provided close-up views and unique perspectives of humpback whales engulfing juvenile salmon at or just below the water surface in Southeast Alaska and Prince William Sound. Although humpback feeding is famous for its flexibility, several stereotyped events were noted in the 47 lunges we analyzed. Engulfment was rapid (mean 2.07 s), and the entrance through which the tongue inverts into the ventral pouch was seen as water rushes in. Cranial elevation was a major contributor to gape, and pouch contraction sometimes began before full gape closure, with reverberating waves indicating rebounding flow of water within the expanded pouch. Expulsion of filtered water began with a small splash at the anterior of the mouth, followed by sustained excurrent flow in the mouth's central or posterior regions. Apart from a splash of rebounding water, water within the mouth was surprisingly turbulence-free during engulfment, but submersion of the whale's head created visible surface whirlpools and vortices which may aggregate prey for subsequent engulfment.

Separating underwater ambient noise from flow noise recorded on stereo acoustic tags attached to marine mammals.

Sound-recording acoustic tags attached to marine animals are commonly used in behavioural studies. Measuring ambient noise is of interest to efforts to understand responses of marine mammals to anthropogenic underwater sound, or to assess their communication space. Noise of water flowing around the tag reflects the speed of the animal, but hinders ambient noise measurement. Here, we describe a correlation-based method for stereo acoustic tags to separate the relative contributions of flow and ambient noise. The uncorrelated part of the noise measured in digital acoustic recording tag (DTAG) recordings related well to swim speed of a humpback whale (Megaptera novaeangliae), thus providing a robust measure of flow noise over a wide frequency bandwidth. By removing measurements affected by flow noise, consistent ambient noise estimates were made for two killer whales (Orcinus orca) with DTAGs attached simultaneously. The method is applicable to any multi-channel acoustic tag, enabling application to a wide range of marine species.

Population structure of humpback whales in the western and central South Pacific Ocean as determined by vocal exchange among populations.

For cetaceans, population structure is traditionally determined by molecular genetics or photographically identified individuals. Acoustic data, however, has provided information on movement and population structure with less effort and cost than traditional methods in an array of taxa. Male humpback whales (Megaptera novaeangliae) produce a continually evolving vocal sexual display, or song, that is similar among all males in a population. The rapid cultural transmission (the transfer of information or behavior between conspecifics through social learning) of different versions of this display between distinct but interconnected populations in the western and central South Pacific region presents a unique way to investigate population structure based on the movement dynamics of a song (acoustic) display. Using 11 years of data, we investigated an acoustically based population structure for the region by comparing stereotyped song sequences among populations and years. We used the Levenshtein distance technique to group previously defined populations into (vocally based) clusters based on the overall similarity of their song display in space and time. We identified the following distinct vocal clusters: western cluster, 1 population off eastern Australia; central cluster, populations around New Caledonia, Tonga, and American Samoa; and eastern region, either a single cluster or 2 clusters, one around the Cook Islands and the other off French Polynesia. These results are consistent with the hypothesis that each breeding aggregation represents a distinct population (each occupied a single, terminal node) in a metapopulation, similar to the current understanding of population structure based on genetic and photo-identification studies. However, the central vocal cluster had higher levels of song-sharing among populations than the other clusters, indicating that levels of vocal connectivity varied within the region. Our results demonstrate the utility and value of using culturally transmitted vocal patterns as a way of defining connectivity to infer population structure. We suggest vocal patterns be incorporated by the International Whaling Commission in conjunction with traditional methods in the assessment of structure.

Estructura Poblacional de las Yubartas en el Centro y Occidente del Sur del Océano Pacífico, Determinada por el Intercambio Vocal entre las Poblaciones Resumen La estructura poblacional de los cetáceos se determina tradicionalmente por medio de genética molecular o individuos identificados fotográficamente. Sin embargo, los datos acústicos han proporcionado información sobre el movimiento y la estructura poblacional de una variedad de taxones con menos esfuerzo y menos costo que los métodos tradicionales. Las yubartas macho (Megaptera novaeangliae) producen una demostración vocal sexual, o canto, en continua evolución que es similar entre todos los machos en una población. La transmisión cultural rápida (la transferencia de información o comportamientos entre co-específicos por medio del aprendizaje social) de diferentes versiones de esta demostración entre poblaciones distintas pero interconectadas en el centro y occidente en la región del sur del Pacífico presenta una manera única para investigar la estructura poblacional con base en las dinámicas de movimiento de la demostración del canto (acústico). Al usar once años de datos, con base en la acústica investigamos a una estructura poblacional de la región al comparar secuencias estereotipadas de cantos entre las poblaciones y los años. Usamos la técnica de la distancia Levenshtein para agrupar poblaciones previamente definidas en grupos (con base vocal) basados en la similitud general de su canto en el espacio y el tiempo. Identificamos a los siguientes grupos vocales perceptibles: grupo occidental, una población al este de Australia; grupo central, poblaciones alrededor de Nueva Caledonia, Tonga y Samoa Americana; y región oriental, ya sea un grupo único o dos grupos, uno alrededor de las Islas Cook y el otro cerca de la Polinesia Francesa. Estos resultados son consistentes con la hipótesis de que cada conjunto reproductivo representa a una población distinta (cada una ocupó un nodo terminal único) en una meta-población, similar al entendimiento actual de la estructura poblacional con base en los estudios genéticos y de foto-identificación. Sin embargo, el grupo vocal central tuvo niveles más altos de cantos compartidos entre las poblaciones que en los otros grupos, lo que indica que los niveles de conectividad vocal variaron dentro de la región. Nuestros resultados demuestran la utilidad y el valor de usar patrones vocales transmitidos culturalmente como una forma de definir la conectividad para inferir la estructura poblacional. Sugerimos que los patrones vocales sean incorporados por la Comisión Internacional de Caza de Ballenas en conjunto con los métodos tradicionales de evaluación de la estructura.

Diel spatio-temporal patterns of humpback whale singing on a high-density breeding ground.

Humpback whale song chorusing dominates the marine soundscape in Hawai'i during winter months, yet little is known about spatio-temporal habitat use patterns of singers. We analysed passive acoustic monitoring data from five sites off Maui and found that ambient noise levels associated with song chorusing decreased during daytime hours nearshore but increased offshore. To resolve whether these changes reflect a diel offshore-onshore movement or a temporal difference in singing activity, data from 71 concurrently conducted land-based theodolite surveys were analysed. Non-calf pods (n = 3082), presumably including the majority of singers, were found further offshore with increasing time of the day. Separately, we acoustically localized 217 nearshore singers using vector-sensors. During the day, distances to shore and minimum distances among singers increased, and singers switched more between being stationary and singing while travelling. Together, these findings suggest that the observed diel trends in humpback whale chorusing off Maui represent a pattern of active onshore-offshore movement of singers. We hypothesize that this may result from singers attempting to reduce intraspecific acoustic masking when densities are high nearshore and avoidance of a loud, non-humpback, biological evening chorus offshore, creating a dynamic of movement of singers aimed at increasing the efficiency of their acoustic display.

Cetacean morbillivirus in Humpback whales' exhaled breath.

The humpback whale (HW; Megaptera novaeangliae) population that seasonally resides along the Brazilian coast concentrates in the Abrolhos Bank (Bahia and Espírito Santo states) for breeding during austral winter and spring. Cetacean morbillivirus (CeMV, Paramyxoviridae family) is currently one of the most significant biological threats to cetaceans worldwide with high infection and mortality rates. CeMV is pleiotropic yet it has special tropism for the respiratory, lymphoid and nervous system and is primarily transmitted by the aerogenous route. A new lineage of CeMV, the Guiana dolphin morbillivirus (GDMV), is known to affect cetaceans off Brazil. GDMV was first detected in a Guiana dolphin (Sotalia guianensis) stranded in the Abrolhos Bank region, in 2010. In addition to pathologic examinations on stranded HW, pathogen survey of free-ranging HW may provide valuable insight into the epidemiology of diseases. We hypothesized that HW in the Brazilian breeding ground could be exposed to CeMV. Thus, in the present study, we investigated the presence of CeMV in exhaled breath condensates (EBC) of HW in the Abrolhos Bank. Overall, 73 samples of EBC from 48 groups of HW were collected during the breeding seasons of 2011 (n = 16) and 2012 (n = 57). One sample failed to have the reference gene amplified and was excluded from the study. CeMV was detected by a RT-qPCR method in 2 EBC samples, representing 2 whale groups. Phylogenetic analysis of partial morbillivirus phosphoprotein gene showed 100% homology to GDMV. Our results show that HW in Brazil are infected by CeMV with a relative prevalence of 4.3% (2/47) and demonstrate the suitability of using EBC and RT-qPCR as a non-invasive tool for CeMV survey in free-ranging whales. This pioneer study provides scientific basis for non-invasive CeMV monitoring of HW, suggests HW may play a role in the dynamics of CeMV and raises concern for potential conservation implications for this species.

Large-scale spatial variabilities in the humpback whale acoustic presence in the Atlantic sector of the Southern Ocean.

Southern Hemisphere humpback whales (Megaptera novaeangliae) inhabit a wide variety of ecosystems including both low- and high-latitude areas. Understanding the habitat selection of humpback whale populations is key for humpback whale stock management and general ecosystem management. In the Atlantic sector of the Southern Ocean (ASSO), the investigation of baleen whale distribution by sighting surveys is temporally restricted to the austral summer. The implementation of autonomous passive acoustic monitoring, in turn, allows the study of vocal baleen whales year-round. This study describes the results of analysing passive acoustic data spanning 12 recording positions throughout the ASSO applying a combination of automatic and manual analysis methods to register humpback whale acoustic activity. Humpback whales were present at nine recording positions with higher acoustic activities towards lower latitudes and the eastern and western edges of the ASSO. During all months, except December (the month with the fewest recordings), humpback whale acoustic activity was registered in the ASSO. The acoustic presence of humpback whales at various locations in the ASSO confirms previous observations that part of the population remains in high-latitude waters beyond austral summer, presumably to feed. The spatial and temporal extent of humpback whale presence in the ASSO suggests that this area may be used by multiple humpback whale breeding populations as a feeding ground.

Deiminated proteins and extracellular vesicles - Novel serum biomarkers in whales and orca.

Peptidylarginine deiminases (PADs) are a family of phylogenetically conserved calcium-dependent enzymes which cause post-translational protein deimination. This can result in neoepitope generation, affect gene regulation and allow for protein moonlighting via functional and structural changes in target proteins. Extracellular vesicles (EVs) carry cargo proteins and genetic material and are released from cells as part of cellular communication. EVs are found in most body fluids where they can be useful biomarkers for assessment of health status. Here, serum-derived EVs were profiled, and post-translationally deiminated proteins and EV-related microRNAs are described in 5 ceataceans: minke whale, fin whale, humpback whale, Cuvier's beaked whale and orca. EV-serum profiles were assessed by transmission electron microscopy and nanoparticle tracking analysis. EV profiles varied between the 5 species and were identified to contain deiminated proteins and selected key inflammatory and metabolic microRNAs. A range of proteins, critical for immune responses and metabolism were identified to be deiminated in cetacean sera, with some shared KEGG pathways of deiminated proteins relating to immunity and physiology, while some KEGG pathways were species-specific. This is the first study to characterise and profile EVs and to report deiminated proteins and putative effects of protein-protein interaction networks via such post-translationald deimination in cetaceans, revealing key immune and metabolic factors to undergo this post-translational modification. Deiminated proteins and EVs profiles may possibly be developed as new biomarkers for assessing health status of sea mammals.

Distinct habitat use strategies of sympatric rorqual whales within a fjord system.

We used ecosystem sampling during systematic surveys and opportunistic focal follows, comparison tests, and random forest models to evaluate fin whale (Balaenoptera physalus) and humpback whale (Megaptera novaeangliae) habitat associations within an inland feeding ground (Kitimat Fjord System, British Columbia, Canada). Though these species are sympatric and share a common prey source, they were attuned to different aspects of the local habitat. The fin whales were associated with habitat properties reminiscent of the open ocean. Humpback whales, in contrast, were associated with features more commonly associated with the inland waters of fjords. Fixed habitat features, such as seafloor depth and distance from the fjord mouth, were the most important predictors of fin whale presence, but fixed and dynamic variables, such as surface properties, predicted humpback whale presence with equal (moderate) success. With the exception of strong salinity gradients for humpback whales, habitat conditions were poor predictors of feeding state. Fin whales practiced a spatially confined, seasonally stable, and thus more predictable use of certain channels within the fjord system. These findings are compatible with site loyal behavior, which is interesting in light of the species' historical, unique use of this fjord system. The relatively lackluster performance of humpback-habitat models, coupled with the importance of oceanographic properties, makes the humpback's habitat use strategy more uncertain. The fact that two sympatric species sharing a common prey source exhibited different habitat use strategies suggests that at least one species was informed by something in addition to prey. Given that the two species are attuned to different aspects of the fjord habitat, their responses to habitat changes, including anthropogenic impacts, would likely be different in both nature and degree. Our findings highlight the value of comparative studies and the complexity of rorqual habitat use, which must be understood in order for critical habitat to be identified and protected.

A Note on Suckling Behavior and Laterality in Nursing Humpback Whale Calves from Underwater Observations.

We investigated nursing behavior on the Hawaiian breeding grounds for first year humpback whale (Megaptera novaeangliae) calves. We observed and video-documented underwater events with nursing behavior from five different whale groups. The observed nursing events include behaviors where a calf positions itself at a 30-45° angle to the midline of the mother's body, with its mouth touching her mammary slit (i.e., suckling position). On two occasions, milk in the water column was recorded in close proximity to a mother/calf pair, and on one occasion, milk was recorded 2.5 min after suckling observed. Nursing events, where the calf was located in the suckling position, were found to be short in duration with a mean of 30.6 s (range 15.0-55.0, standard deviation (SD) = 16.9). All observations of the calf in the suckling position (n = 5, 100%) were with the calf located on the right side of the mother, suggesting a potential for right side laterality preference in the context of nursing behavior. Our study provides insight into mother/calf behaviors from a unique underwater vantage. Results supplement previous accounts of humpback whale nursing in Hawaiian waters, validate mother/calf positioning, document milk in the water column, and introduce the potential for laterality in nursing behavior for humpback whale calves.

Humpback whales feed on hatchery-released juvenile salmon.

Humpback whales are remarkable for the behavioural plasticity of their feeding tactics and the diversity of their diets. Within the last decade at hatchery release sites in Southeast Alaska, humpback whales have begun exploiting juvenile salmon, a previously undocumented prey. The anthropogenic source of these salmon and their important contribution to local fisheries makes the emergence of humpback whale predation a concern for the Southeast Alaska economy. Here, we describe the frequency of observing humpback whales, examine the role of temporal and spatial variables affecting the probability of sighting humpback whales and describe prey capture behaviours at five hatchery release sites. We coordinated twice-daily 15 min observations during the spring release seasons 2010-2015. Using logistic regression, we determined that the probability of occurrence of humpback whales increased after releases began and decreased after releases concluded. The probability of whale occurrence varied among release sites but did not increase significantly over the 6 year study period. Whales were reported to be feeding on juvenile chum, Chinook and coho salmon, with photographic and video records of whales feeding on coho salmon. The ability to adapt to new prey sources may be key to sustaining their population in a changing ocean.

Comportamiento diurno de cetáceos en el Área de Conservación Guanacaste, Costa Rica / Cetacean diurnal behavior in the Guanacaste Conservation Area, Costa Rica

Introducción: Tres especies de cetáceos, el delfín manchado pantropical (Stenella attenuata attenuata), el delfín nariz de botella (Tursiops truncatus) y la ballena jorobada (Megaptera novaeangliae) son un componente importante de la fauna marina del Área de Conservación Guanacaste. Sin embargo, la información biológica básica sobre el comportamiento y la presencia de estas especies sigue siendo escasa. Objetivo: El objetivo de este estudio es determinar el comportamiento diurno de los cetáceos en el sector marino de la ACG y en sus áreas adyacentes. Métodos: Se muestreó el Pacífico norte de Costa Rica desde Bahía Salinas hasta las Islas Murciélago, incluyendo el Golfo de Santa Elena, entre mayo de 2005 y mayo de 2006. Se utilizó el método de escaneo de comportamiento para identificar el comportamiento general de los grupos. Resultados: Se registraron encuentros para tres especies en un total de 36 avistamientos, pero solo con un tiempo considerable para dos especies, S. attenuata y M. novaeangliae. Los delfines manchados exhibieron una amplia variedad de comportamientos en la temporada de lluvias. La búsqueda de alimento (X2 = 29.42; g.l. = 1; P < 0.001) y los viajes (X2 = 38.33; g.l. = 1; P < 0.001) aumentaron significativamente en la época de lluvias. Los comportamientos predominantes para la ballena jorobada fueron la asociación madre-cría, social y de viaje; este último fue mayor en la época de lluvias (X2 = 78.16; g.l. = 1; P < 0.001). Conclusiones: En la zona, los cetáceos presentaron comportamientos como forrajeo y madre-cría, lo que significa que el área puede clasificarse como un hábitat crítico para estas especies de cetáceos, de mayor importancia para especies altamente migratorias como las ballenas jorobadas.

Introduction: Three species of cetaceans, the pantropical spotted dolphin (Stenella attenuata attenuata), the bottlenose dolphin (Tursiops truncatus) and the humpback whale (Megaptera novaeangliae), are important components of the marine fauna of the Guanacaste Conservation Area (ACG). However, basic biological information on the behavior and occurrence of these species remains scarce. Objective: The objective of this study is to determine the diurnal behavior of the three-mentioned species in the ACG. Methods: The north Pacific of Costa Rica, from Bahía Salinas to the Marine Sector of ACG was sampled between May 2005 and May 2006. The behavior scanning method was used to identify the general behavior of the cetacean groups. Results: Encounters were recorded for the three species, in a total of 36 sightings, but only for S. attenuata and M. novaeangliae for considerable periods of time. Stenella attenuata exhibited a wide variety of behaviors, being foraging (X2 = 29.42; d.f. = 1; P < 0.001) and travelling (X2 = 38.33; d.f. = 1; P < 0.001) significantly higher during the rainy season. Predominant behaviors for the humpback whale were mother-calf association, social and travelling; being the latter higher during the rainy season (X2 = 78.16; d.f. = 1; P < 0.001). Conclusions: Behaviors such as foraging and mother-calf were the most representative among species and the most frequent. This suggests that the area can be considered as a critical habitat for these species of cetaceans, with particular importance for highly migratory species such as humpback whales.

Establishment of the first humpback whale fibroblast cell lines and their application in chemical risk assessment.

This paper reports the first successful derivation and characterization of humpback whale fibroblast cell lines. Primary fibroblasts were isolated from the dermal connective tissue of skin biopsies, cultured at 37 °C and 5% CO2 in the standard mammalian medium DMEM/F12 supplemented with 10% fetal bovine serum (FBS). Of nine initial biopsies, two cell lines were established from two different animals and designated HuWa1 and HuWa2. The cells have a stable karyotype with 2n=44, which has commonly been observed in other baleen whale species. Cells were verified as being fibroblasts based on their spindle-shaped morphology, adherence to plastic and positive immunoreaction to vimentin. Population doubling time was determined to be ∼41 h and cells were successfully cryopreserved and thawed. To date, HuWa1 cells have been propagated 30 times. Cells proliferate at the tested temperatures, 30, 33.5 and 37 °C, but show the highest rate of proliferation at 37 °C. Short-term exposure to para,para'-dichlorodiphenyldichloroethylene (p,p'-DDE), a priority compound accumulating in southern hemisphere humpback whales, resulted in a concentration-dependent loss of cell viability. The effective concentration which caused a 50% reduction in HuWa1 cell viability (EC50 value) was approximately six times greater than the EC50 value for the same chemical measured with human dermal fibroblasts. HuWa1 exposed to a natural, p,p'-DDE-containing, chemical mixture extracted from whale blubber showed distinctively higher sensitivity than to p,p'-DDE alone. Thus, we provide the first cytotoxicological data for humpback whales and with establishment of the HuWa cell lines, a unique in vitro model for the study of the whales' sensitivity and cellular response to chemicals and other environmental stressors.

A path reconstruction method integrating dead-reckoning and position fixes applied to humpback whales.

BACKGROUND: Detailed information about animal location and movement is often crucial in studies of natural behaviour and how animals respond to anthropogenic activities. Dead-reckoning can be used to infer such detailed information, but without additional positional data this method results in uncertainty that grows with time. Combining dead-reckoning with new Fastloc-GPS technology should provide good opportunities for reconstructing georeferenced fine-scale tracks, and should be particularly useful for marine animals that spend most of their time under water. We developed a computationally efficient, Bayesian state-space modelling technique to estimate humpback whale locations through time, integrating dead-reckoning using on-animal sensors with measurements of whale locations using on-animal Fastloc-GPS and visual observations. Positional observation models were based upon error measurements made during calibrations. RESULTS: High-resolution 3-dimensional movement tracks were produced for 13 whales using a simple process model in which errors caused by water current movements, non-location sensor errors, and other dead-reckoning errors were accumulated into a combined error term. Positional uncertainty quantified by the track reconstruction model was much greater for tracks with visual positions and few or no GPS positions, indicating a strong benefit to using Fastloc-GPS for track reconstruction. Compared to tracks derived only from position fixes, the inclusion of dead-reckoning data greatly improved the level of detail in the reconstructed tracks of humpback whales. Using cross-validation, a clear improvement in the predictability of out-of-set Fastloc-GPS data was observed compared to more conventional track reconstruction methods. Fastloc-GPS observation errors during calibrations were found to vary by number of GPS satellites received and by orthogonal dimension analysed; visual observation errors varied most by distance to the whale. CONCLUSIONS: By systematically accounting for the observation errors in the position fixes, our model provides a quantitative estimate of location uncertainty that can be appropriately incorporated into analyses of animal movement. This generic method has potential application for a wide range of marine animal species and data recording systems.

Morphology of the eyeball from the Humpback whale (Megaptera novaeangliae).

Aquatic mammals underwent morphological and physiological adaptations due to the transition from terrestrial to aquatic environment. One of the morphological changes regards their vision since cetaceans' eyes are able to withstand mechanical, chemical, osmotic, and optical water conditions. Due to insufficient information about these animals, especially regarding their sense organs, this study aimed to describe the morphology of the Humpback whale (Megaptera novaeangliae) eyeball. Three newborn females, stranded dead on the coast of Sergipe and Bahia, Brazil, were used. Samples were fixed in a 10% formalin solution, dissected, photographed, collected, and evaluated through light and electron microscopy techniques. The Humpback whale sclera was thick and had an irregular surface with mechanoreceptors in its lamina propria. Lens was dense, transparent, and ellipsoidal, consisting of three layers, and the vascularized choroid contains melanocytes, mechanoreceptors, and a fibrous tapetum lucidum. The Humpback whale eyeball is similar to other cetaceans and suggests an adaptation to diving and migration, contributing to the perception of differences in temperature, pressure, and lighting.

Detoxification enzyme activities (CYP1A1 and GST) in the skin of humpback whales as a function of organochlorine burdens and migration status.

The activities of glutathione-s-transferase (GST) and cytochrome P-450 1A1 (CYP1A1) enzymes were measured in freshly extracted epidermis of live-biopsied, migrating, southern hemisphere humpback whales (Megaptera novaeangliae). The two quantified enzyme activities did not correlate strongly with each other. Similarly, neither correlated strongly with any of the organochlorine compound groups previously measured in the superficial blubber of the sample biopsy core, likely reflecting the anticipated low levels of typical aryl-hydrocarbon receptor ligands. GST activity did not differ significantly between genders or between northward (early migration) or southward (late migration) migrating cohorts. Indeed, the inter-individual variability in GST measurements was relatively low. This observation raises the possibility that measured activities were basal activities and that GST function was inherently impacted by the fasting state of the sampled animals, as seen in other species. These results do not support the implementation of CYP1A1 or GST as effective biomarkers of organochlorine contaminant burdens in southern hemisphere populations of humpback whales as advocated for other cetacean species. Further investigation of GST activity in feeding versus fasting cohorts may, however, provide some insight into the fasting metabolism of these behaviourally adapted populations.

Cetacean records along a coastal-offshore gradient in the Vitória-Trindade Chain, western South Atlantic Ocean / Registros de cetáceos ao longo de um gradiente costeiro-pelágico na Cadeia Vitória-Trindade, Oceano Atlântico Sul ocidental

Oceanic waters are difficult to assess, and there are many gaps in knowledge regarding cetacean occurrence. To fill some of these gaps, this article provides important cetacean records obtained in the winter of 2010 during a dedicated expedition to collect visual and acoustic information in the Vitória-Trindade seamounts. We observed 19 groups of cetaceans along a 1300-km search trajectory, with six species being identified: the humpback whale (Megaptera novaeangliae, N = 9 groups), the fin whale (Balaenoptera physalus, N = 1), the Antarctic minke whale (Balaenoptera bonaerensis, N = 1), the rough-toothed dolphin (Steno bredanensis, N = 1), the bottlenose dolphin (Tursiops truncatus, N = 2), and the killer whale (Orcinus orca, N = 1). Most humpback whale groups (N = 7; 78%) were observed in the Vitória-Trindade seamounts, especially the mounts close to the Abrolhos Bank. Only one lone humpback whale was observed near Trindade Island after a search effort encompassing more than 520 km. From a total of 28 acoustic stations, humpback whale songs were only detected near the seamounts close to the Abrolhos Bank, where most groups of this species were visually detected (including a competitive group and groups with calves). The presence of humpback whales at the Trindade Island and surroundings is most likely occasional, with few sightings and low density. Finally, we observed a significant number of humpback whales along the seamounts close to the Abrolhos Bank, which may function as a breeding habitat for this species. We also added important records regarding the occurrence of cetaceans in these mounts and in the Western South Atlantic, including the endangered fin whale.

Águas oceânicas e afastadas da costa são ambientes de difícil acesso e muitas lacunas de conhecimento sobre a ocorrência de cetáceos ainda existem. O presente trabalho fornece importantes registros realizados durante uma expedição dedicada a procurar cetáceos visualmente e acusticamente na Cadeia Vitória-Trindade no inverno de 2010. Foram observados 19 grupos de cetáceos ao longo de 1300 km de esforço, sendo identificadas seis espécies: a baleia-jubarte (Megaptera novaeangliae, N = 9 grupos), a baleia-fin (Balaenoptera physalus, N = 1), um grupo misto de baleia-minke-Antártica (Balaenoptera bonaerensis) e golfinhos-de-dentes-rugosos (Steno bredanensis, N = 1), o golfinho-nariz-de-garrafa (Tursiops truncatus, N = 2) e a orca (Orcinus orca, N = 1). A maioria dos grupos da baleia-jubarte (n=7; 78%) foram observados nos montes marinhos da Cadeia Vitória-Trindade, especialmente os montes próximos do Banco dos Abrolhos. Somente uma baleia-jubarte solitária foi observada próxima da Ilha Trindade, apesar de mais de 520 km percorridos em esforço na região. De um total de 28 estações acústicas, cantos da baleia-jubarte foram detectados somente nos montes próximos ao Banco dos Abrolhos, onde a maioria dos grupos desta espécie foi observada visualmente (incluindo grupos competitivos e fêmeas com filhotes). A presença da baleia-jubarte nos arredores da Ilha de Trindade é provavelmente ocasional, com poucos indivíduos e baixa densidade. Finalmente, um número expressivo de grupos de baleia-jubarte foi observado sobre os montes próximos do Banco dos Abrolhos, que podem funcionar como habitats reprodutivos da espécie. Adicionaram-se ainda importantes registros de cetáceos na Cadeia Vitória-Trindade e no Oceano Atlântico Sul ocidental, incluindo a baleia-fin, uma espécie ameaçada de extinção.

Underwater topography determines critical breeding habitat for humpback whales near Osa Peninsula, Costa Rica: implications for Marine Protected Areas

Migrating humpback whales from northern and southern feeding grounds come to the tropical waters near Osa Peninsula, Pacific of Costa Rica, to reproduce and raise their calves. Planning effective marine protected areas that encompass humpback critical habitats require data about which oceanographic features influence distribution during the breeding period. This study examines the relationship between water depth and ocean floor slope with humpback whale distribution, based on sightings during two breeding seasons (2005 and 2006). Data are from the Southern and Northern subpopulations in the Eastern Tropical Pacific (ETP). Analysis followed the basic principles of the Ecological Niche Factors Analysis (ENFA), where indices of Marginality and Tolerance provide insights on the restrictiveness of habitat use. At a fine scale, physical factors such as water depth and slope define the critical breeding and nursing habitat for M. novaeangliae. Divergence in the subsamples means of depths and slope distribution, with the global mean of the study area in both eco-geographical variables, determine habitat requirements restricted by topographic features such as depths (< 100 m) and slope (< 10%), and locate the key breeding and nursing habitat of the species within the continental shelf domains. Proposed Marine Protected Areas (MPA’s) network plans should consider connectivity of Caño island-Drake Bay and the extension of Corcovado National Park maritime borders. Rev. Biol. Trop. 56 (2): 591-602. Epub 2008 June 30.

Las ballenas jorobadas viajan a aguas tropicales para reproducirse y criar a sus ballenatos. Es importante entender las características oceanográficas que influencian su distribución para lograr una planificación efectiva de áreas marinas protegidas con hábitats críticos para estos cetáceos. Este estudio examina la relación entre la profundidad, la pendiente del suelo oceánico y la distribución de estas ballenas, usando avistamientos del 2005 y 2006 en la costa Pacífica de la Península de Osa, Costa Rica (temporada de reproducción del sur y norte en el Pacífico Tropical Oriental). Usamos Análisis de Factores de Nicho Ecológico (ENFA por sus siglas en inglés), donde los índices de Marginalidad y Tolerancia ilustran las restricciones en uso de hábitat. En una escala local, factores físicos como la profundidad y la pendiente definen el hábitat crítico de reproducción y cría de M. novaeangliae. Las divergencias entre las medias de las sub-muestras y la media global del área de estudio en ambas variables eco-geográficas, determinan las limitaciones en requerimientos de hábitat en aspectos topográficos como la profundidad (>100 m) y la pendiente del fondo (>10%), localizando los hábitat críticos para reproducción y cría dentro de la plataforma continental. Los planes y propuestas para un sistema de áreas marinas protegidas deben de considerar la conectividad de la isla del Caño y la Bahía de Drake, así como la extensión de los límites marinos del Parque Nacional Corcovado.