Rutting vocal display in male impala (Aepyceros melampus) and overlap with alarm context.

BACKGROUND: The rutting vocal display of male impala Aepyceros melampus is unique for its complexity among ruminants. This study investigates bouts of rutting calls produced towards potential mates and rival males by free-ranging male impala in Namibia. In particular, a comparison of male rutting and alarm snorts is conducted, inspired by earlier findings of mate guarding by using alarm snorts in male topi Damaliscus lunatus. RESULTS: Rutting male impala produced 4-38 (13.5 ± 6.5) rutting calls per bout. We analyzed 201 bouts, containing in total 2709 rutting calls of five types: continuous roars produced within a single exhalation-inhalation cycle; interrupted roars including few exhalation-inhalation cycles; pant-roars distinctive by a pant-phase with rapidly alternating inhalations and exhalations; usual snorts lacking any roar part; and roar-snorts starting with a short roar part. Bouts mostly started and ended with usual snorts. Continuous roars were the shortest roars. The average duration of the exhalatory phase was longest in the continuous roars and shortest in the pant-roars. The average fundamental frequency (49.7-51.4 Hz) did not differ between roar types. Vocal tract length, calculated by using measurements of the first four vocal tract resonances (formants), ranged within 381-382 mm in all roar types. In the studied male impala, rutting snorts within bouts of rutting calls were longer and had higher values of the upper quartile in the call spectra than alarm snorts produced towards potential danger. CONCLUSIONS: Additional inhalations during the emission of the interrupted and pant-roars prolong their duration compared to the continuous roars but do not affect the fundamental frequency or the degree of larynx retraction while roaring. Alarm snorts are separated from one another by large intervals, whereas the intervals between rutting snorts within bouts are short. Sometimes, rutting snorts alternate with roars, whereas alarm snorts do not. Therefore, it is not the acoustic structure of individual snorts but the temporal sequence and the occasional association with another call type that defines snorts as either rutting or alarm snorts. The rutting snorts of male impala may function to attract the attention of receptive females and delay their departure from a male's harem or territory.

Vocal phenotype of male rutting roars and genetic markers delineate East European red deer (Cervus elaphus) from Central and West European populations.

This study investigates a population of red deer Cervus elaphus, founded by 10 individuals introduced in the nineteenth century from Germany to the Voronezh region of the European part of Southern Russia and then developed without further introductions. We characterize for the first time the vocal phenotype of the Voronezh red deer male rutting calls in comparison with similar data on the Pannonian (native Central European) and Iberian (native West European) red deer obtained by the authors during preceding studies. In addition, we provide for the first time the genetic data on Pannonian red deer. In Voronezh stags, the number of roars per bout (2.85 ± 1.79) was lower than in Pannonian (3.18 ± 2.17) but higher than in Iberian (2.11 ± 1.71) stags. In Voronezh stags, the duration of main (the longest within bouts) roars was longer (2.46 ± 1.14 s) than in Pannonian (1.13 ± 0.50 s) or Iberian (1.90 ± 0.50 s) stags. The maximum fundamental frequency of main roars was similar between Voronezh (175 ± 60 Hz) and Pannonian (168 ± 61 Hz) but higher in Iberian stags (223 ± 35 Hz). Mitochondrial cytochrome b gene analysis of red deer from the three study populations partially supports the bioacoustical data, of closer similarity between Voronezh and Pannonian populations. In contrast, microsatellite DNA analysis delineates Voronezh red deer from either Pannonian or Iberian red deer. We discuss that population bottlenecking might affect the acoustics of the rutting roars, in addition to genotype.

Savannah roars: The vocal anatomy and the impressive rutting calls of male impala (Aepyceros melampus) - highlighting the acoustic correlates of a mobile larynx.

A retractable larynx and adaptations of the vocal folds in the males of several polygynous ruminants serve for the production of rutting calls that acoustically announce larger than actual body size to both rival males and potential female mates. Here, such features of the vocal tract and of the sound source are documented in another species. We investigated the vocal anatomy and laryngeal mobility including its acoustical effects during the rutting vocal display of free-ranging male impala (Aepyceros melampus melampus) in Namibia. Male impala produced bouts of rutting calls (consisting of oral roars and interspersed explosive nasal snorts) in a low-stretch posture while guarding a rutting territory or harem. For the duration of the roars, male impala retracted the larynx from its high resting position to a low mid-neck position involving an extensible pharynx and a resilient connection between the hyoid apparatus and the larynx. Maximal larynx retraction was 108 mm based on estimates in video single frames. This was in good concordance with 91-mm vocal tract elongation calculated on the basis of differences in formant dispersion between roar portions produced with the larynx still ascended and those produced with maximally retracted larynx. Judged by their morphological traits, the larynx-retracting muscles of male impala are homologous to those of other larynx-retracting ruminants. In contrast, the large and massive vocal keels are evolutionary novelties arising by fusion and linear arrangement of the arytenoid cartilage and the canonical vocal fold. These bulky and histologically complex vocal keels produced a low fundamental frequency of 50 Hz. Impala is another ruminant species in which the males are capable of larynx retraction. In addition, male impala vocal folds are spectacularly specialized compared with domestic bovids, allowing the production of impressive, low-frequency roaring vocalizations as a significant part of their rutting behaviour. Our study expands knowledge on the evolutionary variation of vocal fold morphology in mammals, suggesting that the structure of the mammalian sound source is not always human-like and should be considered in acoustic analysis and modelling.

Ralstonia eutropha H16 in progress: Applications beside PHAs and establishment as production platform by advanced genetic tools.

Ralstonia eutropha strain H16 is a Gram-negative non-pathogenic betaproteobacterium ubiquitously found in soils and has been the subject of intensive research for more than 50 years. Due to its remarkable metabolically versatility, it utilizes a broad range of renewable heterotrophic resources. The substrate utilization range can be further extended by metabolic engineering as genetic tools are available. It has become the best studied "Knallgas" bacterium capable of chemolithoautotrophic growth with hydrogen as the electron donor and carbon dioxide as the carbon source. It also serves as a model organism to study the metabolism of poly(ß-hydroxybutyrate), a polyester which is accumulated within the cells for storage of both carbon and energy. Thermoplastic and biodegradable properties of this polyhydroxyalkanoate (PHA) have attracted much biotechnical interest as a replacement for fossil resource-based plastics. The first applications of R. eutropha aimed at chemolithoautotrophic production of single cell protein (SCP) for food and feed and the synthesis of various PHAs. The complete annotated genome is available allowing systematic biology approaches together with data provided by available omics studies. Besides PHAs, novel biopolymers of 2-hydroxyalkanoates and polythioesters or cyanophycin as well as chemicals such as alcohols, alkanes, alkenes, and further interesting value added chemicals significantly recently extended the range of products synthesized by R. eutropha. High cell density cultivations can be performed without too much effort and the available repertoire of genetic tools is rapidly growing. Altogether, this qualifies R. eutropha strain H16 to become a production platform strain for a large spectrum of products.

Altai pika (Ochotona alpina) alarm calls: individual acoustic variation and the phenomenon of call-synchronous ear folding behavior.

Non-hibernating pikas collect winter food reserves and store them in hay piles. Individualization of alarm calls might allow discrimination between colony members and conspecifics trying to steal food items from a colony pile. We investigated vocal posture, vocal tract length, and individual acoustic variation of alarm calls, emitted by wild-living Altai pikas Ochotona alpina toward a researcher. Recording started when a pika started calling and lasted as long as possible. The alarm call series of 442 individual callers from different colonies consisted of discrete short (0.073-0.157 s), high-frequency (7.31-15.46 kHz), and frequency-modulated calls separated by irregular intervals. Analysis of 442 discrete calls, the second of each series, revealed that 44.34% calls lacked nonlinear phenomena, in 7.02% nonlinear phenomena covered less than half of call duration, and in 48.64% nonlinear phenomena covered more than half of call duration. Peak frequencies varied among individuals but always fitted one of three maxima corresponding to the vocal tract resonance frequencies (formants) calculated for an estimated 45-mm oral vocal tract. Discriminant analysis using variables of 8 calls per series of 36 different callers, each from a different colony, correctly assigned over 90% of the calls to individuals. Consequently, Altai pika alarm calls are individualistic and nonlinear phenomena might further increase this acoustic individualization. Additionally, video analysis revealed a call-synchronous, very fast (0.13-0.23 s) folding, depression, and subsequent re-expansion of the pinna confirming an earlier report of this behavior that apparently contributes to protecting the hearing apparatus from damage by the self-generated high-intensity alarm calls.

Unusually high-pitched neonate distress calls of the open-habitat Mongolian gazelle (Procapra gutturosa) and their anatomical and hormonal predictors.

In neonate ruminants, the acoustic structure of vocalizations may depend on sex, vocal anatomy, hormonal profiles and body mass and on environmental factors. In neonate wild-living Mongolian gazelles Procapra gutturosa, hand-captured during biomedical monitoring in the Daurian steppes at the Russian-Mongolian border, we spectrographically analysed distress calls and measured body mass of 22 individuals (6 males, 16 females). For 20 (5 male, 15 female) of these individuals, serum testosterone levels were also analysed. In addition, we measured relevant dimensions of the vocal apparatus (larynx, vocal folds, vocal tract) in one stillborn male Mongolian gazelle specimen. Neonate distress calls of either sex were high in maximum fundamental frequency (800-900 Hz), but the beginning and minimum fundamental frequencies were significantly lower in males than in females. Body mass was larger in males than in females. The levels of serum testosterone were marginally higher in males. No correlations were found between either body mass or serum testosterone values and any acoustic variable for males and females analysed together or separately. We discuss that the high-frequency calls of neonate Mongolian gazelles are more typical for closed-habitat neonate ruminants, whereas other open-habitat neonate ruminants (goitred gazelle Gazella subgutturosa, saiga antelope Saiga tatarica and reindeer Rangifer tarandus) produce low-frequency (<200 Hz) distress calls. Proximate cause for the high fundamental frequency of distress calls of neonate Mongolian gazelles is their very short, atypical vocal folds (4 mm) compared to the 7-mm vocal folds of neonate goitred gazelles, producing distress calls as low as 120 Hz.

Remarkable vocal identity in wild-living mother and neonate saiga antelopes: a specialization for breeding in huge aggregations?

Saiga antelopes Saiga tatarica tatarica give birth in large aggregations, and offspring follow the herd soon after birth. Herding is advantageous as anti-predator strategy; however, communication between mothers and neonates is strongly complicated in large aggregations. Individual series of nasal and oral contact calls of mother and neonate saiga antelopes were selected from recordings made with automated recording systems placed near the hiding neonates on the saiga breeding grounds in Northern Kazakhstan during synchronized parturitions of 30,000 calving females. We used for comparison of the acoustic structure of nasal and oral contact calls 168 nasal calls of 18 mothers, 192 oral calls of 21 mothers, 78 nasal calls of 16 neonates, and 197 oral calls of 22 neonates. In the oral calls of either mothers or neonates, formant frequencies were higher and the duration was longer than in the nasal calls, whereas fundamental frequencies did not differ between oral and nasal calls. Discriminant function analysis (DFA) based on six acoustic variables, accurately classified individual identity for 99.4% of oral calls of 18 mothers, for 89.3% of nasal calls of 18 mothers, and for 94.4% of oral calls of 18 neonates. The average value of correct classification to individual was higher in mother oral than in mother nasal calls and in mother oral calls than in neonate oral calls; no significant difference was observed between mother nasal and neonate oral calls. Variables mainly responsible for vocal identity were the fundamental frequency and the second and third formants in either mothers or neonates, and in either nasal or oral calls. The high vocal identity of mothers and neonates suggests a powerful potential for the mutual mother-offspring recognition in dense aggregations of saiga antelopes as an important component of their survival strategy.

Engineering the heterotrophic carbon sources utilization range of Ralstonia eutropha H16 for applications in biotechnology.

Ralstonia eutropha H16 is an interesting candidate for the biotechnological production of polyesters consisting of hydroxy- and mercaptoalkanoates, and other compounds. It provides all the necessary characteristics, which are required for a biotechnological production strain. Due to its metabolic versatility, it can convert a broad range of renewable heterotrophic resources into diverse valuable compounds. High cell density fermentations of the non-pathogenic R. eutropha can be easily performed. Furthermore, this bacterium is accessible to engineering of its metabolism by genetic approaches having available a large repertoire of genetic tools. Since the complete genome sequence of R. eutropha H16 has become available, a variety of transcriptome, proteome and metabolome studies provided valuable data elucidating its complex metabolism and allowing a systematic biology approach. However, high production costs for bacterial large-scale production of biomass and biotechnologically valuable products are still an economic challenge. The application of inexpensive raw materials could significantly reduce the expenses. Therefore, the conversion of diverse substrates to polyhydroxyalkanoates by R. eutropha was steadily improved by optimization of cultivation conditions, mutagenesis and metabolic engineering. Industrial by-products and residual compounds like glycerol, and substrates containing high carbon content per weight like palm, soybean, corn oils as well as raw sugar-rich materials like molasses, starch and lignocellulose, are the most promising renewable substrates and were intensively studied.

The power of oral and nasal calls to discriminate individual mothers and offspring in red deer, Cervus elaphus.

BACKGROUND: In most species, acoustical cues are crucial for mother-offspring recognition. Studies of a few species of ungulates showed that potential for individual recognition may differ between nasal and oral contact calls. RESULTS: Vocalizations of 28 hinds and 31 calves of farmed Iberian red deer (Cervus elaphus hispanicus) were examined with discriminant function analyses (DFA) to determine whether acoustic structure of their oral and nasal contact calls encodes information about the caller's identity. Contact calls were elicited by brief separation of individually identified animals by a distance over 10 m or by a bar fence. Both oral and nasal calls of both hinds and calves showed high potential to discriminate individuals. In hinds, individuality was significantly higher in the oral than in the nasal calls, whereas in calves, individuality was equally well expressed in both oral and nasal calls. For calves, the maximum fundamental frequency was higher and the duration was longer in oral calls than in nasal calls. For hinds, the maximum fundamental frequency and the duration were indistinguishable between oral and nasal calls. Compared to the pooled sample of oral and nasal calls, separate oral or nasal call samples provided better classifying accuracy to individual in either hinds or calves. Nevertheless, in both hinds and calves, even in the pooled sample of oral and nasal calls, the degree of individual identity was 2-3 times greater than expected by chance. For hinds that provided calls in both years, cross-validation of calls collected in 2012 with discriminant functions created with calls from 2011 showed a strong decrease of classifying accuracy to individual. CONCLUSIONS: These results suggest different potentials of nasal and oral calls to allow the discrimination of individuals among hinds, but not among red deer calves. The high potential of individual recognition even with the pooled sample of oral and nasal calls allows mother and young to remember only one set of acoustic variables for mutual vocal recognition. Poor between-year stability of individual characteristics of hind oral and nasal calls would require updating keys to individual recognition each calving season.

Vocal group signatures in the goitred gazelle Gazella subgutturosa.

The potential for vocal modification in mammals has recently been of great interest. This study focuses on the potential for vocal matching in juvenile and adolescent goitred gazelles Gazella subgutturosa that were group housed as part of an animal management programme. Two groups of animals (16 and 19 unrelated individuals, respectively) were recorded at two different ages, juvenile and adolescent, regarding 20-25 calls per individual per age; each group was evaluated in a separate year. Vocal similarity of group members compared to non-members was prominent in both ages, but higher in juveniles. Individual identity was prominent in both ages and higher in adolescents. The more prominent vocal indicators of group membership in juveniles could be related to their higher social dependence compared to adolescents. The more individualized calls of adolescents could be a mechanistic consequence of more stable growth at older age. Our results suggest vocal plasticity of goitred gazelles under social influences. These data add to recent evidence about domestic goat kids Capra hircus, suggesting that vocalizations of species that are not capable of imitation are more flexible than previously thought.

Characterization of propionate CoA-transferase from Ralstonia eutropha H16.

In this study, a propionate CoA-transferase (H16_A2718; EC 2.8.3.1) from Ralstonia eutropha H16 (Pct(Re)) was characterized in detail. Glu342 was identified as catalytically active amino acid residue via site-directed mutagenesis. Activity of Pct(Re) was irreversibly lost after the treatment with NaBH4 in the presence of acetyl-CoA as it is shown for all CoA-transferases from class I, thereby confirming the formation of the covalent enzyme-CoA intermediate by Pct(Re). In addition to already known CoA acceptors for Pct Re such as 3-hydroxypropionate, 3-hydroxybutyrate, acrylate, succinate, lactate, butyrate, crotonate and 4-hydroxybutyrate, it was found that glycolate, chloropropionate, acetoacetate, valerate, trans-2,3-pentenoate, isovalerate, hexanoate, octanoate and trans-2,3-octenoate formed also corresponding CoA-thioesters after incubation with acetyl-CoA and Pct(Re). Isobutyrate was found to be preferentially used as CoA acceptor amongst other carboxylates tested in this study. In contrast, no products were detected with acetyl-CoA and formiate, bromopropionate, glycine, pyruvate, 2-hydroxybutyrate, malonate, fumarate, itaconate, ß-alanine, γ-aminobutyrate, levulate, glutarate or adipate as potential CoA acceptor. Amongst CoA donors, butyryl-CoA, crotonyl-CoA, 3-hydroxybutyryl-CoA, isobutyryl-CoA, succinyl-CoA and valeryl-CoA apart from already known propionyl-CoA and acetyl-CoA could also donate CoA to acetate. The highest rate of the reaction was observed with 3-hydroxybutyryl-CoA (2.5 µmol mg⁻¹ min⁻¹). K(m) values for propionyl-CoA, acetyl-CoA, acetate and 3-hydroxybutyrate were 0.3, 0.6, 4.5 and 4.3 mM, respectively. The rather broad substrate range might be a good starting point for enzyme engineering approaches and for the application of Pct(Re) in biotechnological polyester production.

Exploring the Feasibility of Social Skills Programs for Autistic Youth Through Virtual Reality.

Many autistic individuals may desire support and services primarily in the areas of adaptive functioning and social well-being throughout their lifetime. There is a need for community-informed and person-centered social programs targeted for autistic adolescents and adults. New technology, such as virtual reality (VR), is being researched to assess the feasibility of providing therapeutic services to autistic individuals. This qualitative study aims to better understand and explore the perceptions of parents and autistic adolescents regarding social development and thoughts around social skills interventions delivered through technologies such as VR. Eight autistic adolescents between the ages of 12 to 17, and five parents participated across five focus groups. The focus group interviews were semi-structured, and qualitative data analysis was conducted through an open thematic approach and inductive coding process. Seven primary themes with sub-themes were identified throughout the focus group interview discussions: Social Skills of the participating autistic adolescents as reported by their parents and by self-report, Family Dynamics affecting social relationships within the family, Social Skills Goals both groups hoped to develop and improve, the existence and quality of In-Person vs. Online Friendships, Experience/Comfort With Technology, Openness to Virtual Environments, and Concerns for Virtual Environments. Researchers identified these themes across both parent and adolescent focus groups. This study provides guidance to the research community on investigating different innovative approaches for offering a social program that is supported by autistic individuals.

Acoustic complexity of pup isolation calls in Mongolian hamsters: 3-frequency phenomena and chaos.

Studying pup isolation calls of wild rodents provides background for developing new early-life animal models for biomedical research and drug testing. This study discovered a highly complex acoustic phenotype of pup isolation calls in 4-5-day-old Mongolian hamsters Allocricetulus curtatus. We analyzed the acoustic structure of 5,010 isolation calls emitted in the broad range of frequencies (sonic, below 20 kHz, and ultrasonic, from 20 to 128 kHz) by 23 pups during 2-min isolation test trials, 1 trial per pup. In addition, we measured 5 body size parameters and the body weight of each pup. The calls could contain up to 3 independent fundamental frequencies in their spectra, the low (f0), the medium (g0), and the high (h0), or purely consisted of chaos in which the fundamental frequency could not be tracked. By presence/absence of the 3 fundamental frequencies or their combinations and chaos, we classified calls into 6 distinctive categories (low-frequency [LF]-f0, LF-chaos, high-frequency [HF]-g0, HF-h0, HF-g0 + h0, and HF-chaos) and estimated the relative abundance of calls in each category. Between categories, we compared acoustic parameters and estimated their relationship with pup body size index. We discuss the results of this study with data on the acoustics of pup isolation calls reported for other species of rodents. We conclude that such high complexity of Mongolian hamster pup isolation calls is unusual for rodents. Decreased acoustic complexity serves as a good indicator of autism spectrum disorders in knockout mouse models, which makes knockout hamster models prospective new wild animal model of neurodevelopmental disorders.

Relationship between acoustic traits of protesting cries of domestic kittens (Felis catus) and their individual chances for survival.

Domestic cat (Felis catus) mothers may rely on offspring cries to allocate resources in use of individuals with greater chances for survival and sacrifice the weak ones in case of impossibility to raise the entire large litter. Potential victims of this maternal strategy can enhance their chances of survival, by producing vocalizations with traits mimicking those of higher-quality offspring. We compared acoustic traits of 4990 cries produced during blood sampling by 57 two-week-old captive feral kittens (28 males, 29 females); 47 of them survived to 90 days of age and 10 died by reasons not related to traumas or aggression. No relationship was found between acoustic parameters and kitten survival, however, positive relationship was found between survival and body weight. The cries had moderate cues to individuality and lacked cues to sex. Body weight correlated positively with fundamental frequency and negatively with call rate, duration, peak frequency and power quartiles. We discuss that dishonesty of acoustic traits of kitten quality could develop as adaptation for misleading a mother from allocation resources between the weaker and stronger individuals, thus enhancing individual chances for survival for the weaker littermates. Physical constraint, as body weight, may prevent extensive developing the deceptive vocal traits.

Four decades later: The highly conserved repertoire of song types in chaffinch (Fringilla coelebs).

The long-term stability of culturally transmitted traits like bird song has attracted much attention from researchers; however, it remains insufficiently studied up to the present time. This study investigates, by using spectrographic analysis, whether the repertoire of song types recorded at the same locality is persistent at short terms (of four years) and at long terms (of 38 years) in a migratory European continental population of chaffinches. The population song type repertoires recorded in 1978 and 1982 were only slightly different, thus indicating the high repertoire persistence within a few years. There were more differences between repertoires recorded in 1982 and 2020. In total, eight of the 29 song types (28 %) identified in 1982 were not found in 2020, and 5 of the 26 song types (19 %) discovered in 2020 were completely new compared to 1982. All the other song types recorded in 2020 were similar to those recorded in 1982. The frequency of use of these song types in 1982 and 2020 was also similar. By 2020, mainly those song types had disappeared from the population repertoire, which in 1982 were performed by a limited number of males. These data suggest a high long-term persistence of repertoire of song types in male song of a migratory continental population. This long-term stability of the population repertoire was maintained despite significant changes in the habitat structure caused by a massive expansion of bark beetles in 2010-2014.

Comparison of ultrasonic isolation calls of pure-breeding and interspecies hybrid Phodopus dwarf hamster pups.

In mammalian cross-species hybrids, parameters of voice calls, produced by vocal fold vibrations, are intermediate between parental species. Inheritance of ultrasonic calls, produced by whistle mechanism, is unstudied for hybrids. We examined 4000 pup ultrasonic isolation-induced calls for peak power of call fundamental frequency and for call duration in 4-8-day-old captive hamsters of four Study Groups: pure Phodopus sungorus; pure P. campbelli of two populations (Mongolian and Kosh-Agach) and hybrids between male P. sungorus and female P. campbelli (Kosh-Agach). All Study Groups produced two categories of ultrasonic calls: Low-Frequency centered around 41 kHz and High-Frequency centered around 60 kHz, but in different percentages. Between populations, only Low-Frequency calls were shorter and higher-frequency in Mongolian P. campbelli. Between species, only High-Frequency calls were shorter and higher-frequency in P. sungorus. In hybrids, Low-Frequency calls were shorter and lower-frequency than in either parental species, whereas High-Frequency calls were longer and lower-frequency in hybrids than in pure P. sungorus but similar with another parental species. We discuss that interspecific hybridization may give rise to offspring with new properties of ultrasonic calls.

Genetically modified strains of Ralstonia eutropha H16 with ß-ketothiolase gene deletions for production of copolyesters with defined 3-hydroxyvaleric acid contents.

ß-Ketothiolases catalyze the first step of poly(3-hydroxybutyrate) [poly(3HB)] biosynthesis in bacteria by condensation of two acetyl coenzyme A (acetyl-CoA) molecules to acetoacetyl-CoA and also take part in the degradation of fatty acids. During growth on propionate or valerate, Ralstonia eutropha H16 produces the copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [poly(3HB-co-3HV)]. In R. eutropha, 15 ß-ketothiolase homologues exist. The synthesis of 3-hydroxybutyryl-CoA (3HB-CoA) could be significantly reduced in an 8-fold mutant (Lindenkamp et al., Appl. Environ. Microbiol. 76:5373-5382, 2010). In this study, a 9-fold mutant deficient in nine ß-ketothiolase gene homologues (phaA, bktB, H16_A1713, H16_B1771, H16_A1528, H16_B0381, H16_B1369, H16_A0170, and pcaF) was generated. In order to examine the polyhydroxyalkanoate production capacity when short- or long-chain and even- or odd-chain-length fatty acids were provided as carbon sources, the growth and storage behavior of several mutants from the previous study and the newly generated 9-fold mutant were analyzed. Propionate, valerate, octanoate, undecanoic acid, or oleate was chosen as the sole carbon source. On octanoate, no significant differences in growth or storage behavior were observed between wild-type R. eutropha and the mutants. In contrast, during the growth on oleate of a multiple mutant lacking phaA, bktB, and H16_A0170, diminished poly(3HB) accumulation occurred. Surprisingly, the amount of accumulated poly(3HB) in the multiple mutants grown on gluconate differed; it was much lower than that on oleate. The ß-ketothiolase activity toward acetoacetyl-CoA in H16ΔphaA and all the multiple mutants remained 10-fold lower than the activity of the wild type, regardless of which carbon source, oleate or gluconate, was employed. During growth on valerate as a sole carbon source, the 9-fold mutant accumulated almost a poly(3-hydroxyvalerate) [poly(3HV)] homopolyester with 99 mol% 3HV constituents.

Vocal anatomy, tongue protrusion behaviour and the acoustics of rutting roars in free-ranging Iberian red deer stags (Cervus elaphus hispanicus).

Roaring in rutting Iberian red deer stags Cervus elaphus hispanicus is unusual compared to other subspecies of red deer, which radiated from the Iberian refugium after the last glacial maximum. In all red deer stags, the larynx occupies a permanent low mid-neck resting position and is momentarily retracted almost down to the rostral end of the sternum during the production of rutting calls. Simultaneous with the retraction of the larynx, male Iberian red deer pronouncedly protrude the tongue during most of their rutting roars. This poses a mechanical challenge for the vocal tract (vt) and for the hyoid apparatus, as tongue and larynx are strongly pulled in opposite directions. This study (i) examines the vocal anatomy and the acoustics of the rutting roars in free-ranging male C. e. hispanicus; (ii) establishes a potential mechanism of simultaneous tongue protrusion and larynx retraction by applying a two-dimensional model based on graphic reconstructions in single video frames of unrestrained animals; and (iii) advances a hypothesis of evaporative cooling by tongue protrusion in the males of a subspecies of red deer constrained to perform all of the exhausting rutting activities, including acoustic display, in a hot and arid season.

Measuring airborne components of seismic body vibrations in a Middle-Asian sand-dwelling Insectivora species, the piebald shrew (Diplomesodon pulchellum).

Self-produced seismic vibrations have been found for some subterranean rodents but have not been reported for any Insectivora species, although seismic sensitivity has been confirmed for blind sand-dwelling chrysochlorid golden moles. Studying the vocal behaviour of captive piebald shrews, Diplomesodon pulchellum, we documented vibrations, apparently generated by the whole-body wall muscles, from 11 (5 male, 6 female) of 19 animals, placed singly on a drum membrane. The airborne waves of the vibratory drumming were digitally recorded and then analysed spectrographically. The mean frequency of vibration was 160.5 Hz. This frequency matched the periodicity of the deep sinusoidal frequency modulation (159.4 Hz) found in loud screech calls of the same subjects. The body vibration was not related to thermoregulation, hunger-related depletion of energy resources or fear, as it was produced by well-fed, calm animals, at warm ambient temperatures. We hypothesize that in the solitary, nocturnal, digging desert piebald shrew, body vibrations may be used for seismic exploration of substrate density, to avoid energy-costly digging of packed sand for burrowing and foraging. At the same time, the piercing quality of screech calls due to the deep sinusoidal frequency modulation, matching the periodicity of body vibration, may be important for agonistic communication in this species.

Descended and mobile larynx, vocal tract elongation and rutting roars in male goitred gazelles (Gazella subgutturosa Güldenstaedt, 1780).

Similar to male humans, Homo sapiens, the males of a few polygynous ruminants - red deer Cervus elaphus, fallow deer Dama dama and Mongolian gazelle Procapra gutturosa- have a more or less enlarged, low-resting larynx and are capable of additional dynamic vocal tract elongation by larynx retraction during their rutting calls. The vocal correlates of a large larynx and an elongated vocal tract, a low fundamental frequency and low vocal tract resonance frequencies, deter rival males and attract receptive females. The males of the polygynous goitred gazelle, Gazella subgutturosa, provide another, independently evolved, example of an enlarged and low-resting larynx of high mobility. Relevant aspects of the rutting behaviour of territorial wild male goitred gazelles are described. Video and audio recordings served to study the acoustic effects of the enlarged larynx and vocal tract elongation on male rutting calls. Three call types were discriminated: roars, growls and grunts. In addition, the adult male vocal anatomy during the emission of rutting calls is described and functionally discussed using a 2D-model of larynx retraction. The combined morphological, behavioural and acoustic data are discussed in relation to the hypothesis of sexual selection for male-specific deep voices, resulting in convergent features of vocal anatomy in a few polygynous ruminants and in human males.