Vocal Sequences Research Articles

High-precision spatial analysis of mouse courtship vocalization behavior reveals sex and strain differences

Mice display a wide repertoire of vocalizations that varies with sex, strain, and context. Especially during social interaction, including sexually motivated dyadic interaction, mice emit sequences of ultrasonic vocalizations (USVs) of high complexity. As animals of both sexes vocalize, a reliable attribution of USVs to their emitter is essential. The state-of-the-art in sound localization for USVs in 2D allows spatial localization at a resolution of multiple centimeters. However, animals interact at closer ranges, e.g. snout-to-snout. Hence, improved algorithms are required to reliably assign USVs. We present a novel algorithm, SLIM (Sound Localization via Intersecting Manifolds), that achieves a 2–3-fold improvement in accuracy (13.1–14.3 mm) using only 4 microphones and extends to many microphones and localization in 3D. This accuracy allows reliable assignment of 84.3% of all USVs in our dataset. We apply SLIM to courtship interactions between adult C57Bl/6J wildtype mice and those carrying a heterozygous Foxp2 variant (R552H). The improved spatial accuracy reveals that vocalization behavior is dependent on the spatial relation between the interacting mice. Female mice vocalized more in close snout-to-snout interaction while male mice vocalized more when the male snout was in close proximity to the female's ano-genital region. Further, we find that the acoustic properties of the ultrasonic vocalizations (duration, Wiener Entropy, and sound level) are dependent on the spatial relation between the interacting mice as well as on the genotype. In conclusion, the improved attribution of vocalizations to their emitters provides a foundation for better understanding social vocal behaviors.

Bat vocal sequences enhance contextual information independently of syllable order

Many animals, humans included, rely on acoustic vocalizations for communication. The complexity of non-human vocal communication has been under debate one of the main open questions being: What could be the function of multi-syllabic vocal sequences? We address these questions by analyzing fruit-bat vocal communication. We use neural networks to encode the vocalizations, and statistical models to examine the information conveyed by sequences of vocalizations. We show that fruit-bat vocal sequences potentially convey more contextual information than individual syllables, but that the order of the syllables within the sequence is unimportant for context. Specifically, sequences are composed of slightly modified syllables, thus increasing the probability of context-specificity. We note that future behavioral, e.g., playback experiments are needed in order to validate the biological relevance of our statistical results. We hypothesize that such sequences might have served as pre-syntax precursors in the evolution of animal communication.

There You Are! Automated Detection of Indris' Songs on Features Extracted from Passive Acoustic Recordings.

The growing concern for the ongoing biodiversity loss drives researchers towards practical and large-scale automated systems to monitor wild animal populations. Primates, with most species threatened by extinction, face substantial risks. We focused on the vocal activity of the indri (Indri indri) recorded in Maromizaha Forest (Madagascar) from 2019 to 2021 via passive acoustics, a method increasingly used for monitoring activities in different environments. We first used indris' songs, loud distinctive vocal sequences, to detect the species' presence. We processed the raw data (66,443 10-min recordings) and extracted acoustic features based on the third-octave band system. We then analysed the features extracted from three datasets, divided according to sampling year, site, and recorder type, with a convolutional neural network that was able to generalise to recording sites and previously unsampled periods via data augmentation and transfer learning. For the three datasets, our network detected the song presence with high accuracy (>90%) and recall (>80%) values. Once provided the model with the time and day of recording, the high-performance values ensured that the classification process could accurately depict both daily and annual habits of indris' singing pattern, critical information to optimise field data collection. Overall, using this easy-to-implement species-specific detection workflow as a preprocessing method allows researchers to reduce the time dedicated to manual classification.

Network analysis reveals context-dependent structural complexity of social calls in serrate-legged small treefrogs.

Vocal communication plays an important role in survival, reproduction, and animal social association. Birds and mammals produce complex vocal sequence to convey context-dependent information. Vocalizations are conspicuous features of the behavior of most anuran species (frogs and toads), and males usually alter their calling strategies according to ecological context to improve the attractiveness/competitiveness. However, very few studies have focused on the variation of vocal sequence in anurans. In the present study, we used both conventional method and network analysis to investigate the context-dependent vocal repertoire, vocal sequence, and call network structure in serrate-legged small treefrogs Kurixalus odontotarsus. We found that male K. odontotarsus modified their vocal sequence by switching to different call types and increasing repertoire size in the presence of a competitive rival. Specifically, compared with before and after the playback of advertisement calls, males emitted fewer advertisement calls, but more aggressive calls, encounter calls, and compound calls during the playback period. Network analysis revealed that the mean degree, mean closeness, and mean betweenness of the call networks significantly decreased during the playback period, which resulted in lower connectivity. In addition, the increased proportion of one-way motifs and average path length also indicated that the connectivity of the call network decreased in competitive context. However, the vocal sequence of K. odontotarsus did not display a clear small-world network structure, regardless of context. Our study presents a paradigm to apply network analysis to vocal sequence in anurans and has important implications for understanding the evolution and function of sequence patterns.

Slow development of vocal sequences through ontogeny in wild chimpanzees (Pan troglodytes verus).

The development of the unique, hierarchical, and endless combinatorial capacity in a human language requires neural maturation and learning through childhood. Compared with most non-human primates, where combinatorial capacity seems limited, chimpanzees present a complex vocal system comprising hundreds of vocal sequences. We investigated how such a complex vocal system develops and the processes involved. We recorded 10,929 vocal utterances of 98 wild chimpanzees aged 0-55 years, from Taï National Park, Ivory Coast. We developed customized Generalized non-Linear Models to estimate the ontogenetic trajectory of four structural components of vocal complexity: utterance length, diversity, probability of panting (requiring phonation across inhalation and exhalation), and probability of producing two adjacent panted units. We found chimpanzees need 10 years to reach adult levels of vocal complexity. In three variables, the steepest increase coincided with the age of first non-kin social interactions (2-5 years), and plateaued in sub-adults (8-10 years), as individuals integrate into adult social life. Producing two adjacent panted units may require more neuromuscular coordination of the articulators, as its emergence and steepest increase appear later in development. These results suggest prolonged maturational processes beyond those hitherto thought likely in species that do not learn their vocal repertoire. Our results suggest that multifaceted ontogenetic processes drive increases in vocal structural complexity in chimpanzees, particularly increases in social complexity and neuro-muscular maturation. As humans live in a complex social world, empirical support for the "social complexity hypothesis" may have relevance for theories of language evolution. RESEARCH HIGHLIGHTS: Chimpanzees need around 10 years to develop the vocal structural complexity present in the adult repertoire, way beyond the age of emergence of every single vocal unit. Multifaceted ontogenetic processes may drive increases in vocal structural complexity in chimpanzees, particularly increases in social complexity and neuro-muscular maturation. Non-linear increases in vocal complexity coincide with social developmental milestones. Vocal sequences requiring rapid articulatory change emerge later than other vocal sequences, suggesting neuro-muscular maturational processes continue through the juvenile years.

How long are the vocal sequences a passerine bird can memorize? Common song sequences of Radde’s warbler males (Phylloscopus schwarzi) (Passeriformes, Aves)

In most passerine species, an individual bird sings multiple song types to be combined into non-random song sequences. Because of this non-randomness, stereotype sequences of several or even dozens of song types appear in the vocalization of some species. Passerine birds acquire songs through some learning process while imitating other individuals. Song sharing is well known in songbirds and is a consequence of the song learning. Apparently, above the song type level, transitions between song types may be also shared. However, we still do not know exactly, how long are those song sequencies a bird can memorize? We analyze song sequence sharing in Radde’s warbler. Each song of this species consists of a dozen of short notes and lasts 1 s. In many males, the identity of the next song type in a sequence can be predicted on the basis of the previous song type (linear syntax). We found that males can share (i.e. memorize) song sequences from no more than 5 song types. Individual repertoires included up to 40 song types. Therefore, the ability of memorizing song sequences are rather limited in Radde’s warbler, as shared song sequences were rather short in comparison with the total size of the repertoire.

Les lois linguistiques de la brièveté : conformité chez le lémurien chanteur Indri indri

Vocal and gestural sequences of many primate species conform to two principles of compression: the compensation between the length of a construct and that of its constituents (Menzerath-Altmann law) and an inverse relationship between signal length and occurrence (Zipf's law of abbreviation). Although Zipf's law is considered a universal in animal communicative systems, evidence for the adherence of non-human primate vocal behavior to linguistic laws is debated, and information on strepsirrhines is absent. We investigated the vocal behavior of the only singing lemur (Indri indri) to test whether the song of this species shows indication for compression. Because roars have a chaotic structure that prevents the recognition of each individual emission, and long notes are mainly emitted by males, we relied on the core part of the song (i.e., descending phrases, composed of two to six units). Our results indicate that songs conform to both laws. Indeed, shorter phrases are more likely included in the songs, in conformity to Zipf’s law, and unit length decreases with increasing phrase size, in line with Menzerath-Altmann law. Furthermore, despite a sexual dimorphism of both units and phrases duration, these laws characterize the vocal sequences of both sexes. In conclusion, we provide the first evidence for a compromise between signal duration and occurrence in the vocal system of a strepsirrhine species, suggesting that, in primates, selective pressures for compression are more ancestral than previously assumed.

Responses of Hermit Thrush (Catharus guttatus) males to conspecific playback with typical and atypical syntactical patterns

Birdsong exhibits syntactical patterns, wherein singers from a variety of species perform vocal sequences that follow set rules with respect to the order of vocal units (e.g., song types or syllables). However, little is known with certainty regarding the degree to which such patterns contribute to the two main functions of birdsong, mate attraction and territorial defense. The present study investigates the role of syntactical patterns in the songs of the hermit Hermit Thrush (Catharus guttatus), which delivers its song types in a semi-predictable order that cycles up and down the frequency spectrum. A within-subjects playback experiment was conducted to assess the strength of territorial response among males exposed to conspecific song stimuli with and without species-typical patterns. The magnitude of territorial response did not differ with between the two stimulus types, suggesting that the syntactical patterns within Hermit Thrush song do not contribute to territorial defense and aggression. Implications for overall understanding of birdsong syntax, as well as future studies with this species, are discussed.

Auditory pattern discrimination in budgerigars (Melopsittacus undulatus)

Auditory patterns carry information in human speech at multiple levels, including the surface relationships between sounds within words in phonology and the abstract structures of syntax. The sequences of other animal vocalizations, such as birdsong, can also be described as auditory patterns, but few studies have probed how the sequences are perceived at multiple levels. Past work shows that a small parrot species, the budgerigar (Melopsittacus undulatus), exceeds other birds in sequence perception and is even sensitive to abstract structure. But it is not known what level of auditory analysis is dominant in perception or what limits might exist in sensitivity to abstract structure. Here, budgerigars were tested on their ability to discriminate changes in an auditory pattern, AAB, i.e. sound-same different, to ask how they attended to surface relationships among the sounds and the abstract relationships of same/different among the elements. The results show that the budgerigars primarily used surface transitions between the sounds when discriminating the sequences, but were able to use the abstract relationships to a limited extent, largely restricted to two elements. This study provides insight into how budgerigars extract information from conspecific vocalizations and how their capacities compare to human speech perception.

The Ontogeny of Vocal Sequences: Insights from a Newborn Wild Chimpanzee (Pan troglodytes schweinfurthii)

Observations of early vocal behaviours in non-human primates (hereafter primates) are important for direct comparisons between human and primate vocal development. However, direct observations of births and perinatal behaviour in wild primates are rare, and the initial stages of behavioural ontogeny usually remain undocumented. Here, we report direct observations of the birth of a wild chimpanzee (Pan troglodytes schweinfurthii) in Budongo Forest, Uganda, including the behaviour of the mother and other group members. We monitored the newborn’s vocal behaviour for approximately 2 hours and recorded 70 calls. We categorised the vocalisations both qualitatively, using conventional call descriptions, and quantitatively, using cluster and discriminant acoustic analyses. We found evidence for acoustically distinct vocal units, produced both in isolation and in combination, including sequences akin to adult pant hoots, a vocal utterance regarded as the most complex vocal signal produced by this species. We concluded that chimpanzees possess the capacity to produce vocal sequences composed of different call types from birth, albeit in rudimentary forms. Our observations are in line with the idea that primate vocal repertoires are largely present from birth, with fine acoustic structures undergoing ontogenetic processes. Our study provides rare and valuable empirical data on perinatal behaviours in wild primates.

Population-specific call order in chimpanzee greeting vocal sequences

SummaryPrimates rarely learn new vocalizations, but they can learn to use their vocalizations in different contexts. Such “vocal usage learning,” particularly in vocal sequences, is a hallmark of human language, but remains understudied in non-human primates. We assess usage learning in four wild chimpanzee communities of Taï and Budongo Forests by investigating population differences in call ordering of a greeting vocal sequence. Whilst in all groups, these sequences consisted of pant-hoots (long-distance contact call) and pant-grunts (short-distance submissive call), the order of the two calls differed across populations. Taï chimpanzees consistently commenced greetings with pant-hoots, whereas Budongo chimpanzees started with pant-grunts. We discuss different hypotheses to explain this pattern and conclude that higher intra-group aggression in Budongo may have led to a local pattern of individuals signaling submission first. This highlights how within-species variation in social dynamics may lead to flexibility in call order production, possibly acquired via usage learning.

Audience sensitivity in chimpanzee display pant hoots

Audience effects are key in studies of animal social cognition and are typically investigated during directed social interactions. Male chimpanzees, Pan troglodytes, regularly perform aggressive displays in the presence of others, often targeting a specific group member, and combine this agonistic behaviour with acoustic signals. Here, we were interested in whether the production and structure of pant hoots, a long-distance signal, were influenced by audience composition (i.e. presence and absence of specific individuals). We investigated pant hoots produced during displays by adult and subadult males of Budongo Forest, Uganda. We found that males overall called more often when their preferred social partners and females were absent from the party, as well as when more dominant males were present. We then separately analysed the four phases of pant hoots, introduction, build-up, climax and let-down, and found that audience composition and social context could often explain the presence or absence of each phase. In addition, displays were often accompanied by drumming, especially by older males and when male audiences were small. Our study adds to the growing body of literature on audience effects and other social factors and shows their impact on the structure of a sophisticated vocal sequence, which enhances the communicative capacity in a species with limited vocal control.

Chimpanzees produce diverse vocal sequences with ordered and recombinatorial properties

The origins of human language remains a major question in evolutionary science. Unique to human language is the capacity to flexibly recombine a limited sound set into words and hierarchical sequences, generating endlessly new sentences. In contrast, sequence production of other animals appears limited, stunting meaning generation potential. However, studies have rarely quantified flexibility and structure of vocal sequence production across the whole repertoire. Here, we used such an approach to examine the structure of vocal sequences in chimpanzees, known to combine calls used singly into longer sequences. Focusing on the structure of vocal sequences, we analysed 4826 recordings of 46 wild adult chimpanzees from Taï National Park. Chimpanzees produced 390 unique vocal sequences. Most vocal units emitted singly were also emitted in two-unit sequences (bigrams), which in turn were embedded into three-unit sequences (trigrams). Bigrams showed positional and transitional regularities within trigrams with certain bigrams predictably occurring in either head or tail positions in trigrams, and predictably co-occurring with specific other units. From a purely structural perspective, the capacity to organize single units into structured sequences offers a versatile system potentially suitable for expansive meaning generation. Further research must show to what extent these structural sequences signal predictable meanings.

Assessing intra-individual consistency in humpback whale song production using animal-borne acoustic recorders

Studying individual variation in vocal behavior can provide insight into its functions, stability, and mechanisms. Collecting such data at the scale of the individual can be facilitated using animal-borne tags. Here, we use archival suction-cup acoustic recording tags to investigate intra-individual variation in male humpback whale song production. Humpback whale song is a complex and hierarchically structured vocal sequence of 4-7 repeated phrases that are comprised of different units. Repeated songs are termed song sessions. To investigate how consistent song production is within the song session of an individual whale, we deployed suction cup-attached acoustic recording tags on humpback whales in the Hawaiian Islands Humpback Whale National Marine Sanctuary during the 2018–2022 breeding seasons. We analyzed tag data from nine whales to assess intra-individual variation in song structure and syntax as well as differences in the acoustic properties of song units across each song iteration. Each tag contained between 4 and 24 song iterations, and song session recordings varied between 45 minutes and 5 hours. Across individuals, the most variable song iterations occurred at the beginning of a song session. All individuals showed variation in syntax and unit production throughout a session; however, some singers were more consistent than others.

Higher-order sequences of vocal mimicry performed by male Albert's lyrebirds are socially transmitted and enhance acoustic contrast.

Most studies of acoustic communication focus on short units of vocalization such as songs, yet these units are often hierarchically organized into higher-order sequences and, outside human language, little is known about the drivers of sequence structure. Here, we investigate the organization, transmission and function of vocal sequences sung by male Albert's lyrebirds (Menura alberti), a species renowned for vocal imitations of other species. We quantified the organization of mimetic units into sequences, and examined the extent to which these sequences are repeated within and between individuals and shared among populations. We found that individual males organized their mimetic units into stereotyped sequences. Sequence structures were shared within and to a lesser extent among populations, implying that sequences were socially transmitted. Across the entire species range, mimetic units were sung with immediate variety and a high acoustic contrast between consecutive units, suggesting that sequence structure is a means to enhance receiver perceptions of repertoire complexity. Our results provide evidence that higher-order sequences of vocalizations can be socially transmitted, and that the order of vocal units can be functionally significant. We conclude that, to fully understand vocal behaviours, we must study both the individual vocal units and their higher-order temporal organization.

Vocalization-associated respiration patterns: thermography-based monitoring and detection of preparation for calling.

ABSTRACTVocal emission requires coordination with the respiratory system. Monitoring the increase in laryngeal pressure, which is needed for vocal production, allows detection of transitions from quiet respiration to vocalization-supporting respiration. Characterization of these transitions could be used to identify preparation for vocal emission and to examine the probability of it manifesting into an actual vocal production event. Specifically, overlaying the subject's respiration with conspecific calls can highlight events of call initiation and suppression, as a means of signalling coordination and avoiding jamming. Here, we present a thermal imaging-based methodology for synchronized respiration and vocalization monitoring of free-ranging meerkats. The sensitivity of this methodology is sufficient for detecting transient changes in the subject's respiration associated with the exertion of vocal production. The differences in respiration are apparent not only during the vocal output, but also prior to it, marking the potential time frame of the respiratory preparation for calling. A correlation between conspecific calls with elongation of the focal subject's respiration cycles could be related to fluctuations in attention levels or in the motivation to reply. This framework can be used for examining the capability for enhanced respiration control in animals during modulated and complex vocal sequences, detecting ‘failed’ vocalization attempts and investigating the role of respiration cues in the regulation of vocal interactions.

Inhibition of miR-128 Enhances Vocal Sequence Organization in Juvenile Songbirds.

The molecular mechanisms underlying learned vocal communication are not well characterized. This is a major barrier for developing treatments for conditions affecting social communication, such as autism spectrum disorder (ASD). Our group previously generated an activity-dependent gene expression network in the striatopallidal song control nucleus, Area X, in adult zebra finches to identify master regulators of learned vocal behavior. This dataset revealed that the two host genes for microRNA-128, ARPP21 and R3HDM1, are among the top genes whose expression correlates to how much birds sing. Here we examined whether miR-128 itself is behaviorally regulated in Area X and found that its levels decline with singing. We hypothesized that reducing miR-128 during the critical period for vocal plasticity would enhance vocal learning. To test this, we bilaterally injected an antisense miR-128 construct (AS miR-128) or a control scrambled sequence into Area X at post-hatch day 30 (30 d) using sibling-matched experimental and control pupils. The juveniles were then returned to their home cage and raised with their tutors. Strikingly, inhibition of miR-128 in young birds enhanced the organization of learned vocal sequences. Tutor and pupil stereotypy scores were positively correlated, though the correlation was stronger between tutors and control pupils compared to tutors and AS miR-128 pupils. This difference was driven by AS miR-128 pupils achieving higher stereotypy scores despite their tutors’ lower syntax scores. AS miR-128 birds with tutors on the higher end of the stereotypy spectrum were more likely to produce songs with faster tempos relative to sibling controls. Our results suggest that low levels of miR-128 facilitate vocal sequence stereotypy. By analogy, reducing miR-128 could enhance the capacity to learn to speak in patients with non-verbal ASD. To our knowledge, this study is the first to directly link miR-128 to learned vocal communication and provides support for miR-128 as a potential therapeutic target for ASD.

Behavioural context shapes vocal sequences in two anuran species with different repertoire sizes

Acoustic signals in animals serve to convey context-dependent information to receivers. Birds and mammals combine diverse sounds into complex sequences to communicate, but the role of temporal sequencing of signals remains understudied in other taxa. Anuran vocalizations are a prominent feature of their life history, and function in defence of territories and to attract mates. However, there are few data on whether anurans pattern their calls into sequences, and whether temporal sequences convey information about context. Here, we investigated the context-dependent vocal repertoire and the use of vocal sequences by two anuran species belonging to different lineages, comparing frogs vocalizing alone and in the presence of a territorial rival. Using a robust analytical framework, we present evidence that both species modify their vocal sequence structure according to context. Specifically, one species (with a smaller repertoire, from a more basal lineage) appends notes to generate more complex sequences, whereas the other (more recently diverged and with a larger repertoire) shifts to different note types, resulting in different sequences for different contexts. Thus, despite differences in repertoire size, both frog species are capable of adjusting the temporal sequence of vocalizations to communicate in different contexts. Vocal sequences and context-dependent ‘syntax’ may be more common in anurans than previously thought, and our methodology presents a paradigm to study the evolution and function of these complex vocal patterns.

Neural repetition suppression to vocal and non-vocal sounds

Adaptation to the sensory environment is essential in everyday life, to anticipate future events and quickly detect and respond to changes; and to distinguish vocal variations in congeners, for communication. The aim of the current study was to explore the effects of the nature (vocal/non-vocal) of the information to be encoded, on the establishment of auditory regularities. In electrophysiology, neural adaptation is measured by the ‘Repetition Positivity’ (RP), which refers to an increase in positive potential, with the increasing number of repetitions of a same stimulus. The RP results from the combined variation of several ERP components; the P1, the first positivity (∼100 ms) may reflect the onset of repetition effects. We recorded auditory evoked potentials during a roving paradigm in which trains of 4, 8 or 16 repetitions of the same stimulus were presented. Sequences of vocal and non-vocal complex stimuli were delivered, to study the influence of the type of stimulation on the characteristics of the brain responses. The P1 to each train length, and the RP responses were recorded between 90 and 200 ms, reflecting adaptation for both vocal and non-vocal stimuli. RP was not different between vocal and non-vocal sequences (in latency, amplitude and spatial organization) and was found to be similar to that found in previous studies using pure tones, suggesting that the repetition suppression phenomena is somehow independent of the nature of the stimulus. However, results showed faster stabilization of the P1 amplitude for non-vocal stimuli than for vocal stimuli, which require more repetitions. This revealed different dynamics for the establishment of regularity encoding for non-vocal and vocal stimuli, indicating that the richness of vocal sounds may require further processing before full neural adaptation occurs.

Local field potentials in a pre-motor region predict learned vocal sequences.

Neuronal activity within the premotor region HVC is tightly synchronized to, and crucial for, the articulate production of learned song in birds. Characterizations of this neural activity detail patterns of sequential bursting in small, carefully identified subsets of neurons in the HVC population. The dynamics of HVC are well described by these characterizations, but have not been verified beyond this scale of measurement. There is a rich history of using local field potentials (LFP) to extract information about behavior that extends beyond the contribution of individual cells. These signals have the advantage of being stable over longer periods of time, and they have been used to study and decode human speech and other complex motor behaviors. Here we characterize LFP signals presumptively from the HVC of freely behaving male zebra finches during song production to determine if population activity may yield similar insights into the mechanisms underlying complex motor-vocal behavior. Following an initial observation that structured changes in the LFP were distinct to all vocalizations during song, we show that it is possible to extract time-varying features from multiple frequency bands to decode the identity of specific vocalization elements (syllables) and to predict their temporal onsets within the motif. This demonstrates the utility of LFP for studying vocal behavior in songbirds. Surprisingly, the time frequency structure of HVC LFP is qualitatively similar to well-established oscillations found in both human and non-human mammalian motor areas. This physiological similarity, despite distinct anatomical structures, may give insight into common computational principles for learning and/or generating complex motor-vocal behaviors.