Do pingers cause stress in fish? An experimental tank study with European sardine, Sardina pilchardus (Walbaum, 1792) (Actinopterygii, Clupeidae), exposed to a 70 kHz dolphin pinger

Date of acceptance: 06/12/2014 Acknowledgments The study was funded by the Portuguese Ministry of Science (Fundac¸a˜o para a Cieˆncia e Tecnologia– FCT) through a PhD Grant of SG (SFRH/BD/47931/2008). We would like to thank the captain of the purse-seiner (Jose´ Manuel Saveedra) and his crew for facilitating the capture and transport of live fish. Moreover, we want to thank Ana Marc¸alo for suggestions on the experimental design, Manuel Garci for technical advice on underwater video recordings and James Turner from the company Future Oceans for providing technical details on the 70 kHz dolphin pingers. We would also like to acknowledge the scientific advice of Dr. Jose´ Iglesias and the technical and logistic support for the preparation of the laboratory and the materials for tank experiments by Enrique Martı´nez Gonza´lez, Ricardo Pazo´and other staff at the aquaculture facilities of the Spanish Institute for Oceanography (IEO) and the Marine Sciences Station of Toralla (ECIMAT) in Vigo. Furthermore, we are grateful to Francisco de la Granda Grandoso for his practical assistance during the fish tank experiments and to Juan Santos Blanco for helping with statistical analysis. Finally, we would like to thank Pilar Riobo´ Agula, Amelia Fernandez Villamarin, Jose´ Franco Soler, Jose´ Luis Mun˜oz, Angela Benedetti, Marcos Antonio Lopez Patin˜o and Marta Conde Sieira for scientific advice and practical support with cortisol analysis and Rosana Rodrı´guez for preparing histological samples for us.

Mirror self-recognition in the bottlenose dolphin: A case of cognitive convergence

The ability to recognize oneself in a mirror is an exceedingly rare capacity in the animal kingdom. To date, only humans and great apes have shown convincing evidence of mirror self-recognition. Two dolphins were exposed to reflective surfaces, and both demonstrated responses consistent with the use of the mirror to investigate marked parts of the body. This ability to use a mirror to inspect parts of the body is a striking example of evolutionary convergence with great apes and humans.

An account of the voyages undertaken by the order of His present Majesty for making discoveries in the southern hemisphere. And successively performed by Commodore Byron, Captain Wallis, Captain Carteret, and Captain Cook, in the Dolphin, the Swallow, and the Endeavour: drawn up from the journals which were kept by the several commanders, and from the papers of Sir Joseph Banks, bart.

Mode of access: Internet.

S. 39, the International Dolphin Conservation Program Act : hearing before the Subcommittee on Oceans and Fisheries of the Committee on Commerce, Science, and Transportation, United States Senate, One Hundred Fifth Congress, first session, May 14, 1997.

Shipping list no.: 98-0117-P.

Investigation of Group Leadership in a Fission-Fusion Species, the Bottlenose Dolphin

Consistent leadership of group travel by specific individuals has been documented in many animals. Most species exhibiting this type of leadership have relatively stable group membership. Animals using fission-fusion grouping are not expected to use specific leaders because associations would not be frequent. Certain conditions, however, may allow this type of control over group travel to occur. First, a population would need to be small enough to allow regular associations between individuals. Second, leadership may be useful if the environment where the population in question lives is complex and requires learning to access the resources efficiently. To determine whether fission-fusion species existing under these conditions utilize specific individual leadership, I examined a small residential population of bottlenose dolphins (Tursiops truncatus) in the Lower Florida Keys (LFK) where the benthic habitat is highly complex. My goals were to (1) determine whether specific individuals in this population led group travel more often than expected; (2) determine whether certain factors predicted which animals would lead most often and (3) investigate the benefits of leading to leaders and to followers in a fission-fusion society. Multiple types of data were collected to answer questions posed including dolphin behavior (for leadership analyses), fish sampling (to examine dolphin habitat use under leadership), and dolphin biopsy sampling (for genetic analyses). Results of analyses provided strong evidence for consistent leadership in this population. Leaders were female, most were mothers and on average they had larger measures of centrality within the LFK population. Leaders benefited by leading individuals who were more closely related than expected. Followers benefited from efficient access to profitable habitat. Results build on previous leadership research by expanding our knowledge about the type of species in which specific individuals lead and predictors for what types of individuals may lead. Additionally, results provide the first detailed information about benefits group members obtain by both leading and following.^

Could Relatedness Help Explain Why Individuals Lead in Bottlenose Dolphin Groups?

In many species, particular individuals consistently lead group travel. While benefits to followers often are relatively obvious, including access to resources, benefits to leaders are often less obvious. This is especially true for species that feed on patchy mobile resources where all group members may locate prey simultaneously and food intake likely decreases with increasing group size. Leaders in highly complex habitats, however, could provide access to foraging resources for less informed relatives, thereby gaining indirect benefits by helping kin. Recently, leadership has been documented in a population of bottlenose dolphins (Tursiops truncatus) where direct benefits to leaders appear unlikely. To test whether leaders could benefit indirectly we examined relatedness between leader-follower pairs and compared these levels to pairs who associated but did not have leader-follower relationship (neither ever led the other). We found the average relatedness value for leader-follower pairs was greater than expected based on chance. The same was not found when examining non leader-follower pairs. Additionally, relatedness for leader-follower pairs was positively correlated with association index values, but no correlation was found for this measure in non leader-follower pairs. Interestingly, haplotypes were not frequently shared between leader-follower pairs (25%). Together, these results suggest that bottlenose dolphin leaders have the opportunity to gain indirect benefits by leading relatives. These findings provide a potential mechanism for the maintenance of leadership in a highly dynamic fission-fusion population with few obvious direct benefits to leaders.

Immunocytes of the Atlantic bottlenose dolphin (Tursiops truncatus) and West Indian manatee (Trichechus manatus latirostris): Morphologic characterizations and correlations between healthy and disease states under free-ranging and captive conditions

Interest in the health of marine mammals has increased due, in part, to the attention given to human impact on the marine environment. Recent mass strandings of the Atlantic bottlenose dolphin (Tursiops truncatus) and rising mortalities of the endangered Florida manatee (Trichechus manatus latirostris) have raised questions on the extent to which pollution, infectious disease, "stress," and captivity influence the immune system of these animals. This study has provided the first in-depth characterization of immunocytes in the peripheral blood of dolphins (n = 190) and manatees (n = 56). Immunocyte morphology and baseline values were determined in clinically normal animals under free-ranging, stranded and captive living conditions as well as by age and sex. Additionally, immunocyte population dynamics were characterized in sick animals. This was accomplished with traditional cytochemical techniques and new lymphocyte phenotyping methodology which was validated in this study. Traditional cytochemical techniques demonstrated that blood immunocyte morphology and cell numbers are similar to terrestrial mammals with some notable exceptions. The manatee heterophilic granulocyte is a morphologically unique cell and probably functions similarly to the typical mammalian neutrophil. Eosinophils were rarely found in manatees but were uncommonly high in healthy and sick dolphins. Basophils were not identified. Manatees had higher total lymphocyte numbers compared to dolphins and most terrestrial mammals. Lymphocyte subsets identified in healthy animals included T$\rm\sb{h}$, T$\rm\sb{c/s}$, B and NK cells. Dolphin and manatee T and B cell values were higher than those reported in man and most terrestrial mammals. The manatee has extraordinarily high absolute numbers of circulating T$\rm\sb{h}$ cells which suggests an enhanced immunological response capability. With few exceptions, immunocyte types and absolute numbers were not significantly different between free-ranging, stranded and captive categories or between sex and age categories. The evaluation of immunocyte dynamics in various disease states demonstrated a wide variation in cellular responses which provided new insights into innate, humoral and cell-mediated immunity in these species. Additionally, this study demonstrated that lymphocyte phenotyping has diagnostic significance and could be developed into a potential indicator of immunocompetence in both free-ranging and captive dolphin and manatee populations.

Immunocytes of the Atlantic bottlenose dolphin (Tursiops truncatus) and West Indian manatee (Trichechus manatus latirostris) : morphologic characterizations and correlations between healthy and disease states under free-ranging and captive conditions

Interest in the health of marine mammals has increased due, in part, to the attention given to human impact on the marine environment. Recent mass strandings of the Atlantic bottlenose dolphin (Tursiops truncatus) and rising mortalities of the endangered Florida manatee (Trichechus manatus latirostris) have raised questions on the extent to which pollution, infectious disease, "stress," and captivity influence the immune system of these animals. This study has provided the first in-depth characterization of immunocytes in the peripheral blood of dolphins (n=180) and manatees (n=56). Immunocyte morphology and baseline values were determined in clinically normal animals under free-ranging, stranded and captive living conditions as well as by age and sex. Additionally, immuocyte population dynamics were characterized in sick animals. This was accomplished with traditional cytochemical techniques and new lymphocyte phenotyping methodology which was validated in this study. Traditional cytochemical techniques demonstrated that blood immunocyte morphology and cell numbers are similar to terrestrial mammals with some notable exceptions. The manatee heterophilic granulocyte is a morphologically unique cell and probably functions similarly to the typical mammalian neutrophil. Eosinophils were rarely found in manatees but were uncommonly high in healthy and sick dolphins. Basophils were not identified. Manatees had higher total lymphocyte numbers compared to dolphins and most terrestrial mammals. Lymphocyte subsets identified in healthy animals included Th, Tes, B and NK cells. Dolphin and manatee T and B cell values were higher than those reported in man and most terrestrial mammals. The manatee has extraordinarily high absolute numbers of circulating Th cells which suggests an enhanced immunological response capability. With few exceptions, immunocyte types and absolute numbers were not significantly different between free-ranging, stranded and captive categories or between sex and age categories. The evaluation of immunocyte dynamics in various disease states demonstrated a wide variation in cellular responses which provided new insights into innate, humoral and cell-mediated immunity in these species. Additionally, this study demonstrated that lymphocyte phenotyping has diagnostic significance and could be developed into a potential indicator of immunocompetence in both free-ranging and captive dolphin and manatee populations.

Exploring the applicability of biological and socioeconomic tools in developing EAFM plans for data absent areas : Spinner dolphin EAFM for Kalpitiya, Sri Lanka

Acknowledgements University of Aberdeen, UK and Bay of Bengal Large Marine Ecosystems (BOBLME) project are acknowledged for partial funding of this research.

Soundscape Ecology of Hawaiian Spinner Dolphin Resting Bays

Sound is a key sensory modality for Hawaiian spinner dolphins. Like many other marine animals, these dolphins rely on sound and their acoustic environment for many aspects of their daily lives, making it is essential to understand soundscape in areas that are critical to their survival. Hawaiian spinner dolphins rest during the day in shallow coastal areas and forage offshore at night. In my dissertation I focus on the soundscape of the bays where Hawaiian spinner dolphins rest taking a soundscape ecology approach. I primarily relied on passive acoustic monitoring using four DSG-Ocean acoustic loggers in four Hawaiian spinner dolphin resting bays on the Kona Coast of Hawai‛i Island. 30-second recordings were made every four minutes in each of the bays for 20 to 27 months between January 8, 2011 and March 30, 2013. I also utilized concomitant vessel-based visual surveys in the four bays to provide context for these recordings. In my first chapter I used the contributions of the dolphins to the soundscape to monitor presence in the bays and found the degree of presence varied greatly from less than 40% to nearly 90% of days monitored with dolphins present. Having established these bays as important to the animals, in my second chapter I explored the many components of their resting bay soundscape and evaluated the influence of natural and human events on the soundscape. I characterized the overall soundscape in each of the four bays, used the tsunami event of March 2011 to approximate a natural soundscape and identified all loud daytime outliers. Overall, sound levels were consistently louder at night and quieter during the daytime due to the sounds from snapping shrimp. In fact, peak Hawaiian spinner dolphin resting time co-occurs with the quietest part of the day. However, I also found that humans drastically alter this daytime soundscape with sound from offshore aquaculture, vessel sound and military mid-frequency active sonar. During one recorded mid-frequency active sonar event in August 2011, sound pressure levels in the 3.15 kHz 1/3rd-octave band were as high as 45.8 dB above median ambient noise levels. Human activity both inside (vessels) and outside (sonar and aquaculture) the bays significantly altered the resting bay soundscape. Inside the bays there are high levels of human activity including vessel-based tourism directly targeting the dolphins. The interactions between humans and dolphins in their resting bays are of concern; therefore, my third chapter aimed to assess the acoustic response of the dolphins to human activity. Using days where acoustic recordings overlapped with visual surveys I found the greatest response in a bay with dolphin-centric activities, not in the bay with the most vessel activity, indicating that it is not the magnitude that elicits a response but the focus of the activity. In my fourth chapter I summarize the key results from my first three chapters to illustrate the power of multiple site design to prioritize action to protect Hawaiian spinner dolphins in their resting bays, a chapter I hope will be useful for managers should they take further action to protect the dolphins.

Dolphin in lower Shark River Slough

This material is based upon work supported by the National Science Foundation through the Florida Coastal Everglades Long-Term Ecological Research program under Cooperative Agreements #DBI-0620409 and #DEB-9910514. This image is made available for non-commercial or educational use only.