Measuring rhythms of vocal interactions: a proof of principle in harbour seal pups.

Rhythmic patterns in interactive contexts characterize human behaviours such as conversational turn-taking. These timed patterns are also present in other animals, and often described as rhythm. Understanding fine-grained temporal adjustments in interaction requires complementary quantitative methodologies. Here, we showcase how vocal interactive rhythmicity in a non-human animal can be quantified using a multi-method approach. We record vocal interactions in harbour seal pups (Phoca vitulina) under controlled conditions. We analyse these data by combining analytical approaches, namely categorical rhythm analysis, circular statistics and time series analyses. We test whether pups' vocal rhythmicity varies across behavioural contexts depending on the absence or presence of a calling partner. Four research questions illustrate which analytical approaches are complementary versus orthogonal. For our data, circular statistics and categorical rhythms suggest that a calling partner affects a pup's call timing. Granger causality suggests that pups predictively adjust their call timing when interacting with a real partner. Lastly, the ADaptation and Anticipation Model estimates statistical parameters for a potential mechanism of temporal adaptation and anticipation. Our analytical complementary approach constitutes a proof of concept; it shows feasibility in applying typically unrelated techniques to seals to quantify vocal rhythmic interactivity across behavioural contexts. This article is part of a discussion meeting issue 'Face2face: advancing the science of social interaction'.

Vocal tract allometry in a mammalian vocal learner.

Acoustic allometry occurs when features of animal vocalisations can be predicted from body size measurements. Despite this being considered the norm, allometry sometimes breaks, resulting in species sounding smaller or larger than expected for their size. A recent hypothesis suggests that allometry-breaking mammals cluster into two groups: those with anatomical adaptations to their vocal tracts and those capable of learning new sounds (vocal learners). Here, we tested which mechanism is used to escape from acoustic allometry by probing vocal tract allometry in a proven mammalian vocal learner, the harbour seal (Phoca vitulina). We tested whether vocal tract structures and body size scale allometrically in 68 young individuals. We found that both body length and body mass accurately predict vocal tract length and one tracheal dimension. Independently, body length predicts vocal fold length while body mass predicts a second tracheal dimension. All vocal tract measures are larger in weaners than in pups and some structures are sexually dimorphic within age classes. We conclude that harbour seals do comply with anatomical allometric constraints. However, allometry between body size and vocal fold length seems to emerge after puppyhood, suggesting that ontogeny may modulate the anatomy-learning distinction previously hypothesised as clear cut. We suggest that seals, and perhaps other species producing signals that deviate from those expected from their vocal tract dimensions, may break allometry without morphological adaptations. In seals, and potentially other vocal learning mammals, advanced neural control over vocal organs may be the main mechanism for breaking acoustic allometry.

Vocal plasticity in harbour seal pups.

Vocal plasticity can occur in response to environmental and biological factors, including conspecifics' vocalizations and noise. Pinnipeds are one of the few mammalian groups capable of vocal learning, and are therefore relevant to understanding the evolution of vocal plasticity in humans and other animals. Here, we investigate the vocal plasticity of harbour seals (Phoca vitulina), a species with vocal learning abilities observed in adulthood but not puppyhood. To evaluate early mammalian vocal development, we tested 1-3 weeks-old seal pups. We tailored noise playbacks to this species and age to induce seal pups to shift their fundamental frequency (f0), rather than adapt call amplitude or temporal characteristics. We exposed individual pups to low- and high-intensity bandpass-filtered noise, which spanned-and masked-their typical range of f0; simultaneously, we recorded pups' spontaneous calls. Unlike most mammals, pups modified their vocalizations by lowering their f0 in response to increased noise. This modulation was precise and adapted to the particular experimental manipulation of the noise condition. In addition, higher levels of noise induced less dispersion around the mean f0, suggesting that pups may have actively focused their phonatory efforts to target lower frequencies. Noise did not seem to affect call amplitude. However, one seal showed two characteristics of the Lombard effect known for human speech in noise: significant increase in call amplitude and flattening of spectral tilt. Our relatively low noise levels may have favoured f0 modulation while inhibiting amplitude adjustments. This lowering of f0 is unusual, as most animals commonly display no such f0 shift. Our data represent a relatively rare case in mammalian neonates, and have implications for the evolution of vocal plasticity and vocal learning across species, including humans. This article is part of the theme issue 'Voice modulation: from origin and mechanism to social impact (Part I)'.

Neuroanatomy of the grey seal brain: bringing pinnipeds into the neurobiological study of vocal learning.

Comparative animal studies of complex behavioural traits, and their neurobiological underpinnings, can increase our understanding of their evolution, including in humans. Vocal learning, a potential precursor to human speech, is one such trait. Mammalian vocal learning is under-studied: most research has either focused on vocal learning in songbirds or its absence in non-human primates. Here, we focus on a highly promising model species for the neurobiology of vocal learning: grey seals (Halichoerus grypus). We provide a neuroanatomical atlas (based on dissected brain slices and magnetic resonance images), a labelled MRI template, a three-dimensional model with volumetric measurements of brain regions, and histological cortical stainings. Four main features of the grey seal brain stand out: (i) it is relatively big and highly convoluted; (ii) it hosts a relatively large temporal lobe and cerebellum; (iii) the cortex is similar to that of humans in thickness and shows the expected six-layered mammalian structure; (iv) there is expression of FoxP2 present in deeper layers of the cortex; FoxP2 is a gene involved in motor learning, vocal learning, and spoken language. Our results could facilitate future studies targeting the neural and genetic underpinnings of mammalian vocal learning, thus bridging the research gap from songbirds to humans and non-human primates. Our findings are relevant not only to vocal learning research but also to the study of mammalian neurobiology and cognition more in general. This article is part of the theme issue 'Vocal learning in animals and humans'.

HEMATOLOGY AND SERUM BIOCHEMISTRY OF HARBOR SEAL (PHOCA VITULINA) PUPS AFTER REHABILITATION IN THE NETHERLANDS.

Hematology and serum biochemistry profiles are used to evaluate the health status of animals ongoing rehabilitation. The aim of this project was to develop blood and biochemistry ranges for harbor seal pups (Phoca vitulina) after rehabilitation; thus, 22 different blood parameters in 60 animals were tested before release. The second goal was to test for differences due to sex, stranding location, body condition at admission, and presence or absence of umbilical cord. The alanine aminotransferase, ALT (or glutamate pyruvate transaminase, GPT), (ALT-GPT) differed significantly (P bq = 0.00851) between sexes. Lower leukocyte counts and higher liver enzyme values were the most remarkable findings when comparing the results of this study to other published data. This is the first study to report blood reference ranges for harbor seal pups in the Dutch Wadden Sea after rehabilitation.

RETROSPECTIVE STUDY OF SURGICAL TREATMENT OF REFRACTIVE OSTEOMYELITIS AND INFECTIOUS ARTHRITIS IN THE FLIPPERS OF SEALS IN THE NETHERLANDS.

Amputation surgery in pinniped rehabilitation centers is a feasible procedure when animals are presented with open fractures, osteomyelitis, and/or infectious arthritis of the flippers that appear to be refractory to medical treatment. From 2011 to 2017, the Sealcentre Pieterburen in The Netherlands admitted 3,775 seals for rehabilitation. Of these, 37 individuals presented clinical and radiologic signs of bone abnormalities indicative of osteomyelitis or infectious arthritis refractory to medical treatment. Seven cases resulted in euthanasia, and 30 cases underwent amputation surgery. The surgical procedure involved amputation of part of a flipper (24; two animals twice) or of a complete flipper (eight). All procedures were done under general anesthesia except one that was performed with local anesthesia, and all 30 animals were released. In two cases, the osteomyelitis presented with the rare Totenlade phenomenon, a sequestrum surrounded by new periosteal bone formation. The purpose of this retrospective study was to evaluate the outcome of the operative treatment of osteomyelitis and infectious arthritis in the flippers of harbor (Phoca vitulina) and grey seals (Halichoerus grypus) during this 6-yr period.