Domestication dramatically changes behaviour, including communication, as seen in the case of dogs (Canis familiaris) and wolves (Canis lupus). We tested the hypothesis that domestication may affect an ancient, shared communication form of canids, the howling which seems to have higher individual variation in dogs: the perception and usage of howls may be affected by the genetic relatedness of the breeds to their last common ancestor with wolves ('root distance') and by other individual features like age, sex, and reproductive status. We exposed 68 purebred dogs to wolf howl playbacks and recorded their responses. We identified an interaction between root distance and age on the dogs' vocal and behavioural responses: older dogs from more ancient breeds responded longer with howls and showed more stress behaviours. Our results suggest that domestication impacts vocal behaviour significantly: disintegrating howling, a central, species-specific communication form of canids and gradually eradicating it from dogs' repertoire.

Density estimation is a key issue in wildlife management but is particularly challenging and labour-intensive for elusive species. Recently developed approaches based on remotely collected data and capture-recapture models, though representing a valid alternative to more traditional methods, have found little application to species with limited morphological variation. We implemented a camera trap capture-recapture study to survey wolf packs in a 560-km2 area of Central Italy. Individual recognition of focal animals (alpha) in the packs was possible by relying on morphological and behavioural traits and was validated by non-invasive genotyping and inter-observer agreement tests. Two types (Bayesian and likelihood-based) of spatially explicit capture-recapture (SCR) models were fitted on wolf pack capture histories, thus obtaining an estimation of pack density in the area.

Monitoring wolves (Canis lupus) is a difficult and often expensive task due to high mobility, pack dynamic, shyness and nocturnal activity of this species. Wolves communicate acoustically through howling, within pack and with packs of the neighbourhood. A wolf howl is a low-frequency vocalization that can be transmitted over long distances and thus it can be used for monitoring. Elicited howling survey is a current method to monitor wolves in different areas all over the world. Elicited howling, however, may be invasive to residential wolf packs and could create possible negative reactions from the human population. Here we show that it is possible to detect wolves by recording spontaneous howling events. We measured the sound pressure level of wolf howls by captive individuals and we further found that elicited howling may be recorded and clearly identified up to a distance of 3 km. We finally conducted a non-invasive acoustic detection of wolves in a free-ranging population. The use of passive sound recorders may provide a powerful non-invasive tool for future wolf monitoring and could help to establish sustainable management plans for this species.

Vocal divergence within species often corresponds to morphological, environmental, and genetic differences between populations. Wolf howls are long-range signals that encode individual, group, and subspecies differences, yet the factors that may drive this variation are poorly understood. Furthermore, the taxonomic division within the Canis genus remains contended and additional data are required to clarify the position of the Himalayan, North African, and Indian wolves within Canis lupus. We recorded 451 howls from the 3 most basal wolf lineages—Himalayan C. lupus chanco—Himalayan haplotype, North African C. lupus lupaster, and Indian C. lupus pallipes wolves—and present a howl acoustic description within each clade. With an additional 619 howls from 7 Holarctic subspecies, we used a random forest classifier and principal component analysis on 9 acoustic parameters to assess whether Himalayan, North African, and Indian wolf howls exhibit acoustic differences compared to each other and Holarctic wolf howls. Generally, both the North African and Indian wolf howls exhibited high mean fundamental frequency (F0) and short duration compared to the Holarctic clade. In contrast, the Himalayan wolf howls typically had lower mean F0, unmodulated frequencies, and short howls compared to Holarctic wolf howls. The Himalayan and North African wolves had the most acoustically distinct howls and differed significantly from each other and to the Holarctic wolves. Along with the influence of body size and environmental differences, these results suggest that genetic divergence and/or geographic distance may play an important role in understanding howl variation across subspecies.

Wolf choruses (Canis lupus) are complex, multicomponent signals, composed by a series of different vocalizations emitted by a pack. Although howls, the main component, have been highly studied, poor attention has been drawn upon the other vocalizations of the chorus. In this study, we investigate the structure of the chorus by means of the analysis and the quantification of the different components, taking advantage both of the digital sound recording and analysis, and of the modern statistical methodologies. We provide for the first time a detailed, objective description of the types of call emitted during the wolf howlings, combining spectrographic examinations, spectral analyses and automated classifications, with the aim to identify different types of call. Our results show that wolf choruses have a rich, complex structure, that reveals six other types of call, to be added to those howls already described in literature. Wolf choruses are typically composed by other three different types of calls: the bark, i.e. relatively long calls characterized by low frequencies and the presence of harsh components (deterministic chaos); the whimper, characterized by a harmonic structure and a very short duration; and the growl, a call with a noisy structure, low frequencies but relative long duration. Although further investigations are necessary to understand the meaning of the different calls, this research provides a basis for those studies that aim to compare wolves and other canids vocal behaviour.

ABSTRACTWith an average male body mass of 320 kg, the wapiti, Cervus canadensis, is the largest extant species of Old World deer (Cervinae). Despite this large body size, male wapiti produce whistle-like sexual calls called bugles characterised by an extremely high fundamental frequency. Investigations of the biometry and physiology of the male wapiti's relatively large larynx have so far failed to account for the production of such a high fundamental frequency. Our examination of spectrograms of male bugles suggested that the complex harmonic structure is best explained by a dual-source model (biphonation), with one source oscillating at a mean of 145 Hz (F0) and the other oscillating independently at an average of 1426 Hz (G0). A combination of anatomical investigations and acoustical modelling indicated that the F0 of male bugles is consistent with the vocal fold dimensions reported in this species, whereas the secondary, much higher source at G0 is more consistent with an aerodynamic whistle produced as air flows rapidly through a narrow supraglottic constriction. We also report a possible interaction between the higher frequency G0 and vocal tract resonances, as G0 transiently locks onto individual formants as the vocal tract is extended. We speculate that male wapiti have evolved such a dual-source phonation to advertise body size at close range (with a relatively low-frequency F0 providing a dense spectrum to highlight size-related information contained in formants) while simultaneously advertising their presence over greater distances using the very high-amplitude G0 whistle component.

Wolves (Canis lupus) in Italy represent a relict west European population. They are classified as vulnerable by IUCN, though have increased in number and expanded their range in recent decades. Here we use 17 years of monitoring data (from 1993 to 2010) collected in a mountainous region of central Italy (Arezzo, Tuscany) in an ecological niche-based model (MaxEnt) to characterize breeding sites (i.e. the areas where pups were raised) within home ranges, as detected from play-back responses. From a suite of variables related to topography, habitat and human disturbance we found that elevation and distance to protected areas were most important in explaining the locality of wolf responses. Rendezvous sites (family play-back response sites) typically occurred between 800 and 1200 m a.s.l., inside protected areas, and were usually located along mountain chains distant from human settlements and roads. In these areas human disturbance is low and the densities of ungulates are typically high. Over recent years, rendezvous sites have occurred closer to urban areas as the wolf population has continued to expand, despite the consequent human disturbance. This suggests that undisturbed landscapes may be reaching their carrying capacity for wolves. This, in turn, may lead to the potential for increased human-wolf interactions in future. Applying our model, both within and beyond the species' current range, we identify sites both within the current range and also further afield, that the species could occupy in future. Our work underlines the importance of the present protected areas network in facilitating the recolonisation by wolves. Our projections of suitability of sites for future establishment as the population continues to expand could inform planning to minimize future wolf-human conflicts.

Introduction: Monitoring large carnivores is a central issue in conservation biology. The wolf (Canis lupus) is the most studied large carnivore in the world. After a massive decline and several local extinctions, mostly due to direct persecutions, wolves are now recolonizing many areas of their historical natural range. One of the main monitoring techniques is the howling survey, which is based on the wolves' tendency to use vocalisations to mark territory ownership in response to howls of unknown individuals. In most cases wolf howling sessions are useful for the localisation of the pack, but they provide only an aural estimation of the chorus size. We tested and present a new bioacoustic approach to estimate chorus size by recording wolves' replies and visualising choruses through spectrograms and spectral envelopes. To test the methodology, we compared: a) the values detected by visual inspections with the true chorus size to test for accuracy; b) the bioacoustic estimations of a sample of free-ranging wolves' replies developed by different operators to test for precision of the method; c) the aural field estimation of chorus size of a sample of free-ranging wolves' replies with the sonogram analysis of the same recordings to test for difference between methods.Results: Visual inspection of the chorus by spectrogram and spectrum proved to be useful in determining the number of concurrent voices in a wolf chorus. Estimations of chorus size were highly correlated with the number of wolves counted in a pack, and 92 % of 29 known chorus sizes were recognized by means of bioacoustic analysis. On the basis of spectrographic evidence, it was also possible to identify up to seven concurrent vocalisations in a chorus of nine wolves. Spectral analysis of 37 free ranging wolves' replies showed a high correlation between the chorus size estimations of the different operators (92.8 %), but a low correlation with the aural estimation (59.2 %).Conclusions: Wolf howling monitoring technique could be improved by recording wolves' replies and by using bioacoustic tools such as spectrograms and spectral envelopes to determine the size of the wolf chorus. Compared with other monitoring techniques (i.e., genetic analysis), bioacoustic analysis requires widely available informatic tools (i.e., sound recording set of devices and sound analysis software) and a low budget. Information obtained by means of chorus analysis can also be combined with that provided by other techniques. Moreover, howls can be recorded and stored in audio file format with a good resolution (i.e. in "Wave" format), thus representing a useful tool for future listening and investigations, which can be countlessly employed without risks of time deterioration.

Mesola red deer are the only native red deer population remaining in peninsular Italy (Mattioli and Ferretti 2014; Zachos et al. 2014). Their peculiar morphological aspects and genetic makeup have earned them the qualification of Evolutionary Significant Unit (ESU), and the status of subspecies has recently been proposed. Despite this, their behaviour remains poorly documented. Because vocal behaviour, including rutting vocalisations, can provide useful phylogenetic signals (Cap et al. 2008), we present the first description of the main sexually selected male call. We find that roars of Mesola red deer are characterised by a relatively low average fundamental frequency (79 Hz) compared to that of most continental red deer populations, confirming the phenotypic originality of Mesola red deer and strengthening the case for its conservation. We also report that roars contain cues to identity, which could support individual recognition tools for monitoring this endangered population.

We provide a detailed description of the rutting vocalisations of free-ranging male Iberian deer (Cervus elaphus hispanicus, Hilzheimer 1909), a geographically isolated and morphologically differentiated subspecies of red deer Cervus elaphus. We combine spectrographic examinations, spectral analyses and automated classifications to identify different call types, and compare the composition of the vocal repertoire with that of other red deer subspecies. Iberian stags give bouts of roars (and more rarely, short series of barks) that are typically composed of two different types of calls. Long Common Roars are mostly given at the beginning or at the end of the bout, and are characterised by a high fundamental frequency (F0) resulting in poorly defined formant frequencies but a relatively high amplitude. In contrast, Short Common Roars are typically given in the middle or at the end of the bout, and are characterised by a lower F0 resulting in relatively well defined vocal tract resonances, but low amplitude. While we did not identify entirely Harsh Roars (as described in the Scottish red deer subspecies (Cervus elaphus scoticus)), a small percentage of Long Common Roars contained segments of deterministic chaos. We suggest that the evolution of two clearly distinct types of Common Roars may reflect divergent selection pressures favouring either vocal efficiency in high pitched roars or the communication of body size in low-pitched, high spectral density roars highlighting vocal tract resonances. The clear divergence of the Iberian red deer vocal repertoire from those of other documented European red deer populations reinforces the status of this geographical variant as a distinct subspecies.

The high variability of vocalization is due to their function, the habitat where they evolved and the physical constraints of the emitters. The aim of this study were to analyse the acoustic behaviour of two large terrestrial mammals, wolf ( Canis lupus ) and red deer ( Cervus elaphus ), in relation to resources maintenance and mating systems. Firstly, I analysed the Italian wolf howl. I found two forms of howl; both types are uttered within the lowest frequencies of the wolf’s vocal range, confirming the acoustic adaptation hypothesis, and thus, that low frequencies are more useful for long distance communication. Moreover, I found a group vocal signature into the packs. This characteristic is constant both within biological season and between two consecutive years. Secondly, I analysed the red deer mating calls. I described the vocal repertoire of Iberian red deer ( C. e. hispanicus ) males and I quantified variation of the roar of three free ranging subspecies. Iberian red deer repertoire consists of four call types and presents the “short common roar”, absent in the repertoire of the others subspecies. Among red deer subspecies exists a strong vocal divergence: C.e. hippelaphus present the highest intra population variability, followed by C. e. corsicanus and C. e. hispanicus . Between subspecies, the highest bioacoustic differences were found between C. e. hispanicus and C. e. corsicanus while this last population and C.e. hippelaphus are the most related subspecies.

Acoustic communication conveys a variety of information that is a helpful tool for animal conservation. The wolf is an elusive species, which can be detected through the howls that individuals emit. In this study we investigated the acoustic features of wild wolf pack howls from five locations in the province of Arezzo, Italy. We tested the hypothesis that each group had a distinctive vocal signature. Our results showed that these wolf packs emitted howls with significantly distinctive acoustic structures. We hypothesized that group-specific vocal signatures require temporal stability to be functional. Indeed, we did not find any statistical differences in howls collected from the same location during the same season or for 2 consecutive years. We suggest that the acoustic features of howls can be used to distinguish wolf packs in the wild.

Italian wolf howls are described for the first time from observations between 2003-2008 of a population living in eastern Tuscany, central Italy. A sample of 37 howls selected among single responses and 128 howls included in the choruses of 7 free ranging packs was recorded and analysed. The mean fundamental frequency of the howls ranged between 274-908 Hz. Two main structures recognised by means of multivariate explorative analysis, in particular Principal Component and Cluster Analysis, were ascribed to breaking and flat howls. Discriminant Function Analysis was applied to the recognised groups with the aim to find a general rule for classification. Howls with different features were correctly assigned to the groups obtained by explorative analysis in 95.8% of cases. The analysis of the variables characterising the structure of the howls suggests that maximum frequency and range of fundamental frequency are the most important parameters for classification, while duration does not appear to play any significant role.