Research Note
Evaluation of ultrasonic vocalizations in common marmosets (Callithrix jacchus) as a potential indicator of welfare Jaco Bakker, DVM1, Tessa J.M. van Nijnatten2, Annet L. Louwerse1, Guus Baarends2,3, Saskia S. Arndt, PhD4 & Jan A.M. Langermans, PhD1
© 2014 Nature America, Inc. All rights reserved.
The vocal repertoire in common marmosets (Callithrix jacchus) has been assumed to consist not only of vocalizations audible to humans but also of ultrasonic vocalizations (USVs). The use of USVs to socially indicate distress has not been evaluated in this species, however. The authors analyzed the ultrasonic vocal repertoire of the common marmoset under normal housing conditions, under various experimental manipulations intended to elicit positive or negative emotional responses and during stressful experiences including blood draw and exposure to a perceived predator. Analysis of the recordings showed that marmosets produced vocalizations with ultrasonic components as part of their normal vocal repertoire, but these vocalizations all have audible components as well. Only 4 of the 13 types of vocalizations had ultrasonic components. These ultrasonic components were not reliably associated with responses to different experimental manipulations, suggesting that they are not used to indicate pain, discomfort or distress.
npg
Intra-species communication of many species, including flying squirrels1, bornean frogs2,3, rats4, voles5, minks6 and non-human primates7–21, consists of vocalizations with frequencies in the ultrasonic range ( > 20 kHz) as well as in the range audible to humans (20 Hz–20 kHz). New World monkeys have vocal repertoires comprising both audible and ultrasonic frequencies8,22; common marmosets (Callithrix jacchus) can hear sounds with frequencies 4 months old. In three of these groups, the breeding female was on birth control (Implanon, Organon n.v., Oss, the Netherlands). Five family groups consisted of relatives of the same sex (e.g., father and sons). We recorded the sound in each cage once for 30 min daily between 9:00 a.m. and 12:00 p.m., excluding weekends, for 1 month. We opened the plastic front doors of the cage that was being recorded, leaving the doors of all the other cages closed. Vocalizations in response to experimental manipulations We recorded vocalizations during various experi mental manipulations expected to elicit positive or negative emotional states in the animals, as determined by our own experience and by the literature (Table 1). We carried out these experiments in the home cages of six adult male marmosets (2–3 y of age), all housed in the same room in the experimental facility with no other marmosets present. Two of these marmosets that had been pair-housed were separated due to incompatibility and were temporarily singly housed during the experiment. The other four marmosets were pair-housed. During each trial, marmosets in each cage were exposed to the experimental manipulation and recorded one at a time for 20 min each. During trials in which pair-housed marmosets were recorded, we determined which of the two animals produced the vocalization, and we determined the social rank order of the marmosets (dominant versus subordinate) by visual observation. During Trial A, we recorded baseline vocalizations of the marmosets in undisturbed conditions while engaging in normal behavior (Table 1). During Trial B, we offered one marshmallow (Frisia mini mallows; Astra Faam BV, Harlingen, the Netherlands) by hand to each of the marmosets. Marshmallows are commonly used to reward these marmosets during training, and therefore they were expected to have a positive emotional response to the marshmallow. During Trial C, we presented a blue leather welder glove (Liberty Glove Inc., City of Industry, CA) to the marmosets by hand. This handling glove is used to restrain the marmosets when needed (e.g., during biannual vaccination); therefore, they were expected to have a negative emotional response to the glove. One singly housed marmoset was not included in Trial C, because it was sedated for use in another study at the time. During Trial D, we visually separated the pair-housed marmosets (n = 4) by placing an iron plate horizontally in the cage to divide it into two separate compartments, each containing one of the two marmosets. The separation was expected to elicit a negative emotional response in the marmosets. www.labanimal.com
Research Note
npg
© 2014 Nature America, Inc. All rights reserved.
TABLE 1 | Experimental manipulations during recording of vocalizations Emotional response expected
Trial
Manipulation
A
None
n/a
Four (four pair-housed and two singly housed marmosets)
B
Presentation of marshmallows
Positive
Four (four pair-housed and two singly housed marmosets)
C
Presentation of blue catching glove
Negative
Three (four pair-housed and one singly housed marmosets)
D
Visual separation
Negative21,39
Two (four pair-housed marmosets)
E
Introduction of unfamiliar animals
Negative24
Two (two pair-housed and one singly housed marmosets)
During Trial E, we removed three of the marmosets (two pair-housed and one singly housed) from the room and left the other three marmosets (two pairhoused and one singly housed) in the room. Then, we introduced three unfamiliar males of the same age into the room. The introduction of the unfamiliar males was also expected to elicit a negative emotional response in the marmosets.
their response to a neutral novel object (cow). One of the two toys was placed inside the cage for a period of 5–7 min. Two groups were first presented with the toy snake and later with the toy cow, whereas the third group was first presented with the cow and then with the snake. A one-month intervening period separated the two presentations. Only one presentation of each toy was made to each group.
Vocalizations during blood draw We recorded the vocalizations of nine male marmosets and nine female marmosets (3–8 y of age) during blood sampling without sedation. These marmosets were housed as same-sex pairs; one randomly selected individual from each pair was sampled. Marmosets were transported individually to an adjacent, temperaturecontrolled (24 °C) observation room. After recording began, a caretaker restrained the conscious marmoset using blue leather welder gloves (Liberty Glove Inc., City of Industry, CA) while a second caretaker inserted a 26-gauge needle (0.5-in Sterican Hypodermic Needle; B. Braun, Melsungen, Germany) percutaneously into the saphenous vein33 and collected a 200-ml blood sample for use in another study. No topical analgesics were applied prior to blood sampling. If a first attempt at blood sampling was not successful, a second attempt was made. After blood sampling was completed, we stopped the audio recording.
Audio recording and data analysis We recorded behaviors and audible vocalizations using a digital camcorder (Sony DCR-SR72E; Tokyo, Japan). The frequency threshold for audible vocalizations was set at 20 kHz. We analyzed the USVs using dedicated Sonotrack software as well as the opensource software programs mp3DirectCut (mpesch3 audio tools, Frankfurt, Germany), SYRINX (Higher Concept Software, Reading, UK) and Avisoft SASLab Lite (Avisoft Bioacoustics, Glienicke, Germany). For each experimental condition, we determined the total number of audible vocalizations, the vocalization rate and the presence of ultrasonic frequency components for each group. For the blood draw experiment and in the imitation predator experiment, we used the MannWhitney U test to determine the median and the 25% and 75% interquartile range (IQR) for USV production rates. We considered P values 20 kHz (ref. 1). Because the presence of higher harmonics in the vocalizations, which extend into the ultrasonic range, may be a consequence of more intense (energetic) calling, the impact of vocal intensity should be considered. Greater energy in the fundamental (‘audible’) component may make the ultrasonic components easier to detect. Unfortunately, we were unable to determine the energy in the fundamental components without USV and the fundamental components with USV. More research is needed for detailed analysis of spectral energy, fundamental and harmonic frequencies in the vocalizations of marmosets. We found that audible calls with ultrasonic components were produced by marmosets under normal conditions and also in response to restraint stress and confrontation with a perceived predator. Because we were not able to distinguish between the production of USVs in normal and stressful conditions, USV production cannot be used as a reliable indicator of stress in marmosets. Whether the presence of ultrasonic frequencies in the vocalizations of marmosets has any biological relevance remains a salient question. Common marmosets have a hearing range of up to 36 kHz (ref. 23), and since production of USVs is energyconsuming, it is likely that vocalizations made at these high frequencies are purposeful1. For example, USVs might be used to detect objects and obstacles, to measure distances or to locate prey. It might serve as a private channel of communication that cannot be intercepted by predators or prey. Additionally, the ultrasonic component might contain information about the sender (as in bird song), although we did not detect any individual USV signatures in this study. Although the current data do not clarify the function of USV production in marmosets, our observations that a limited range of audible vocalizations produced by marmosets contain ultrasonic components offer new opportunities to study the biological relevance of marmoset vocalizations. Acknowledgments We thank Herma van der Wiel for critical reading of the manuscript and Donna Devine and Thea de Koning for editing the manuscript. We also thank Henk van Westbroek for optimizing the graphs and figures. This study was funded in part by a grant from the EUPRIM-Net Project. COMPETING FINANCIAL INTERESTS The authors declare no competing financial interests. Received 3 January 2014; accepted 7 May 2014 Published online at http://www.labanimal.com/
LAB ANIMAL
1. Murrant, M.N. et al. Ultrasonic vocalizations emitted by flying squirrels. PLoS ONE 8, e73045 (2013). 2. Arch, V.S., Grafe, T.U., Gridi-Papp, M. & Narins, P.M. Pure ultrasonic communication in an endemic Bornean frog. PLoS ONE 4, e5413 (2009). 3. Arch, V.S. & Narins, P.M. “Silent” signals: Selective forces acting on ultrasonic communication systems in terrestrial vertebrates. Anim. Behav. 76, 1423–1428 (2008). 4. Brudzynski, S.M. Communication of adult rats by ultrasonic vocalization: biological, sociobiological, and neuroscience approaches. ILAR J. 50, 43–50 (2009). 5. Yu, P. et al. The effects of repeated early deprivation on ultrasonic vocalizations and ontogenetic development in mandarin vole pups. Behav. Processes 88, 162–167 (2011). 6. Clausen, K.T., Malmkvist, J. & Surlykke, A. Ultrasonic vocalizations of kits during maternal kit-retrieval in farmed mink, Mustela vison. Appl. Anim. Behav. Sci. 114, 582–592 (2008). 7. Bezerra, B.M. & Souta, A. Structure and usage of the vocal repertoire of Callithrix jacchus. Int. J. Primatol. 29, 671–701 (2008). 8. Epple, G. Comparative studies on vocalization in marmoset monkeys (Hapalidae). Folia Primatol. (Basel) 8, 1–40 (1968). 9. Goldman, J.A. Acoustic communication in the common marmoset monkey (Callithrix jacchus jacchus) measurements of transmissions and responses. PhD Dissertation, York University, Toronto, Canada (2000). 10. Jones, B.S. Vocal behaviour of the common marmoset: structure and function of selected calls. Primate Eye 53, 22–23 (1993). 11. Mendes, S.L., Vielliard, J.M.E. & De Marco, P. in The Smallest Anthropoids: The Marmoset/Callimico Radiation (Developments in Primatology: Progress and Prospects) (eds. Ford, S.M., Porter, L.M. & Davis, L.C.) 63–84 (Springer, New York, NY, 2009). 12. Miller, C.T., Mandel, K. & Wang, X. The communicative content of the common marmoset phee call during antiphonal calling. Am. J. Primatol. 72, 974–980 (2010). 13. Morrill, R.J., Thomas, A.W., Schiel, N., Souto, A. & Miller, C.T. The effect of habitat acoustics on common marmoset vocal signal transmission. Am. J. Primatol. 75, 904–916 (2013). 14. Norcross, J.L., Newman, J.D. & Cofrancesco, L.M. Context and sex differences exist in the acoustic structure of phee calls by newly-paired common marmosets (Callithrix jacchus). Am. J. Primatol. 49, 165–181 (1999). 15. Pistorio, A.L., Vintch, B. & Wang, X. Acoustic analysis of vocal development in a New World primate, the common marmoset (Callithrix jacchus). J. Acoust. Soc. Am. 120, 1655–1670 (2006). 16. Ramsier, M.A. et al. Primate communication in the pure ultrasound. Biol. Lett. 8, 508–511 (2012). 17. Stevenson, M.F. & Poole, T.B. An ethogram of the common marmoset (Calithrix jacchus jacchus): general behavioural repertoire. Anim. Behav. 24, 428–451 (1976). 18. Stevenson, M.F. & Rylands, A.B. in Ecology and Behavior of Neotropical Primates vol. 2. (eds. Mittermeier, R.A., Rylands, A.B., Coimbra-Filho, A.F. & Fonseca, G.A.B.) 131–222 (World Wildlife Fund, Washington, DC, 1988). 19. Watson, C.F. & Caldwell, C.A. Neighbor effects in marmosets: social contagion of agonism and affiliation in captive Callithrix jacchus. Am. J. Primatol. 72, 549–558 (2010). 20. Yamaguchi, C., Izumi, A. & Nakamura, K. Temporal rules in vocal exchanges of phees and trills in common marmosets (Callithrix jacchus). Am. J. Primatol. 71, 617–622 (2009). 21. Yamaguchi, C., Izumi, A. & Nakamura, K. Time course of vocal modulation during isolation in common marmosets (Callithrix jacchus). Am. J. Primatol. 72, 681–688 (2010). 22. Anderson, J.W. The production of ultrasonic sounds by laboratory rats and other mammals. Science 119, 808–809 (1954).
Volume 43, No. 9 | SEPTEMBER 2014 319
23. Osmanski, M.S. & Wang, X. Measurement of absolute auditory thresholds in the common marmoset (Callithrix jacchus). Hear. Res. 277, 127–133 (2011). 24. Gerber, P., Schnell, C.R. & Anzenberger, G. Behavioral and cardiophysiological responses of common marmosets (Callithrix jacchus) to social and environmental changes. Primates 43, 201–216 (2002). 25. Abbott, D.H., Barnett, D.K., Colman, R.J., Yamamoto, M.E. & Schultz-Darken, N.J. Aspects of common marmoset basic biology and life history important for biomedical research. Comp. Med. 53, 339–350 (2003). 26. Mansfield, K. Marmoset models commonly used in biomedical research. Comp. Med. 53, 383–392 (2003). 27. Burman, O.H.P., Ilyat, A., Jones, G. & Mendl, M. Ultrasonic vocalizations as indicators of welfare for laboratory rats (Rattus norvegicus). Appl. Anim. Behav. Sci. 104, 116–129 (2007). 28. Kikusui, T., Nishizawa, D., Takeuchi, Y. & Mori, Y. Conditioned fear-related ultrasonic vocalizations are emitted as an emotional response. J. Vet. Med. Sci. 65, 1299–1305 (2003). 29. Takahashi, N., Kashino, M. & Hironaka, N. Structure of rat ultrasonic vocalizations and its relevance to behavior. PLoS ONE 5, e14115 (2010). 30. Wöhr, M. & Schwarting, R.K. Ultrasonic communication in rats: can playback of 50-kHz calls induce approach behavior? PLoS ONE 2, e1365 (2007). 31. Portfors, C.V. Types and functions of ultrasonic vocalizations in laboratory rats and mice. J. Am. Assoc. Lab. Anim. Sci. 46, 28–34 (2007). 32. Williams, W.O., Riskin, D.K. & Mott, A.K. Ultrasonic sound as an indicator of acute pain in laboratory mice. J. Am. Assoc. Lab. Anim. Sci. 47, 8–10 (2008). 33. Jagessar, S.A. et al. Overview of models, methods, and reagents developed for translational autoimmunity research in the common marmoset (Callithrix jacchus). Exp. Anim. 62, 159–171 (2013).
34. Cagni, P., Sampaio, A.C., Ribeiro, N.B. & Barros, M. Immediate, but no delayed, behavioral response to a snake model by captive black tufted-ear marmosets. Behav. Processes 87, 241–245 (2011). 35. Clara, E., Tommasi, L. & Rogers, L.J. Social mobbing calls in common marmosets (Callithrix jacchus): effects of experience and associated cortisol levels. Anim. Cogn. 11, 349–358 (2008). 36. Emile, N. & Barros, M. Recognition of a 3D snake model and its 2D photographic image by captive black tufted-ear marmosets (Callithrix penicillata). Anim. Cogn. 12, 725–732 (2009). 37. Kaplan, G., Pines, M.K. & Rogers, L.J. Stress and stress reduction in common marmosets. Appl. Anim. Behav. Sci. 137, 175–182 (2012). 38. Norcross, J.L. & Newman, J.D. Context and gender-specific differences in the acoustic structure of common marmoset (Callithrix jacchus) phee calls. Am. J. Primatol. 30, 37–54 (1993). 39. Boere, V., Paludob, G.R., Pianta, G., Canale, C. & Tomaz, C. Effects of novelty, isolation stress, and environmental enrichment on some haematological parameters in marmosets (Callithrix penicillata). Comp. Biochem. Phys. Pharm. Tox. 119, 261–274 (2003). 40. Ely, A., Freer, A., Windle, C. & Ridley, R.M. Assessment of cage use by laboratory-bred common marmosets (Callithrix jacchus). Lab. Anim. 32, 427–433 (1998). 41. Searcy, Y.M. & Caine, N.G. Hawk calls elicit alarm and defensive reactions in captive Geoffroy’s marmosets (Callithrix geoffroyi). Folia Primatol. (Basel) 74, 115–125 (2003). 42. Winter, M. in Biology and Behaviour of Marmosets: Proceedings of the Marmoset Workshop, Göttingen, West Germany, 2–5 September 1977 (eds. Rothe, H., Wolters, H.J. & Hearn, J.P.) 127–139 (H. Rothe, Göttingen, Germany, 1978).
npg
© 2014 Nature America, Inc. All rights reserved.
Research Note
320 Volume 43, No. 9 | SEPTEMBER 2014
www.labanimal.com
Evaluation of ultrasonic vocalizations in common marmosets (Callithrix jacchus) as a potential indicator of welfare Jaco Bakker, DVM1, Tessa J.M. van Nijnatten2, Annet L. Louwerse1, Guus Baarends2,3, Saskia S. Arndt, PhD4 & Jan A.M. Langermans, PhD1
© 2014 Nature America, Inc. All rights reserved.
The vocal repertoire in common marmosets (Callithrix jacchus) has been assumed to consist not only of vocalizations audible to humans but also of ultrasonic vocalizations (USVs). The use of USVs to socially indicate distress has not been evaluated in this species, however. The authors analyzed the ultrasonic vocal repertoire of the common marmoset under normal housing conditions, under various experimental manipulations intended to elicit positive or negative emotional responses and during stressful experiences including blood draw and exposure to a perceived predator. Analysis of the recordings showed that marmosets produced vocalizations with ultrasonic components as part of their normal vocal repertoire, but these vocalizations all have audible components as well. Only 4 of the 13 types of vocalizations had ultrasonic components. These ultrasonic components were not reliably associated with responses to different experimental manipulations, suggesting that they are not used to indicate pain, discomfort or distress.
npg
Intra-species communication of many species, including flying squirrels1, bornean frogs2,3, rats4, voles5, minks6 and non-human primates7–21, consists of vocalizations with frequencies in the ultrasonic range ( > 20 kHz) as well as in the range audible to humans (20 Hz–20 kHz). New World monkeys have vocal repertoires comprising both audible and ultrasonic frequencies8,22; common marmosets (Callithrix jacchus) can hear sounds with frequencies 4 months old. In three of these groups, the breeding female was on birth control (Implanon, Organon n.v., Oss, the Netherlands). Five family groups consisted of relatives of the same sex (e.g., father and sons). We recorded the sound in each cage once for 30 min daily between 9:00 a.m. and 12:00 p.m., excluding weekends, for 1 month. We opened the plastic front doors of the cage that was being recorded, leaving the doors of all the other cages closed. Vocalizations in response to experimental manipulations We recorded vocalizations during various experi mental manipulations expected to elicit positive or negative emotional states in the animals, as determined by our own experience and by the literature (Table 1). We carried out these experiments in the home cages of six adult male marmosets (2–3 y of age), all housed in the same room in the experimental facility with no other marmosets present. Two of these marmosets that had been pair-housed were separated due to incompatibility and were temporarily singly housed during the experiment. The other four marmosets were pair-housed. During each trial, marmosets in each cage were exposed to the experimental manipulation and recorded one at a time for 20 min each. During trials in which pair-housed marmosets were recorded, we determined which of the two animals produced the vocalization, and we determined the social rank order of the marmosets (dominant versus subordinate) by visual observation. During Trial A, we recorded baseline vocalizations of the marmosets in undisturbed conditions while engaging in normal behavior (Table 1). During Trial B, we offered one marshmallow (Frisia mini mallows; Astra Faam BV, Harlingen, the Netherlands) by hand to each of the marmosets. Marshmallows are commonly used to reward these marmosets during training, and therefore they were expected to have a positive emotional response to the marshmallow. During Trial C, we presented a blue leather welder glove (Liberty Glove Inc., City of Industry, CA) to the marmosets by hand. This handling glove is used to restrain the marmosets when needed (e.g., during biannual vaccination); therefore, they were expected to have a negative emotional response to the glove. One singly housed marmoset was not included in Trial C, because it was sedated for use in another study at the time. During Trial D, we visually separated the pair-housed marmosets (n = 4) by placing an iron plate horizontally in the cage to divide it into two separate compartments, each containing one of the two marmosets. The separation was expected to elicit a negative emotional response in the marmosets. www.labanimal.com
Research Note
npg
© 2014 Nature America, Inc. All rights reserved.
TABLE 1 | Experimental manipulations during recording of vocalizations Emotional response expected
Trial
Manipulation
A
None
n/a
Four (four pair-housed and two singly housed marmosets)
B
Presentation of marshmallows
Positive
Four (four pair-housed and two singly housed marmosets)
C
Presentation of blue catching glove
Negative
Three (four pair-housed and one singly housed marmosets)
D
Visual separation
Negative21,39
Two (four pair-housed marmosets)
E
Introduction of unfamiliar animals
Negative24
Two (two pair-housed and one singly housed marmosets)
During Trial E, we removed three of the marmosets (two pair-housed and one singly housed) from the room and left the other three marmosets (two pairhoused and one singly housed) in the room. Then, we introduced three unfamiliar males of the same age into the room. The introduction of the unfamiliar males was also expected to elicit a negative emotional response in the marmosets.
their response to a neutral novel object (cow). One of the two toys was placed inside the cage for a period of 5–7 min. Two groups were first presented with the toy snake and later with the toy cow, whereas the third group was first presented with the cow and then with the snake. A one-month intervening period separated the two presentations. Only one presentation of each toy was made to each group.
Vocalizations during blood draw We recorded the vocalizations of nine male marmosets and nine female marmosets (3–8 y of age) during blood sampling without sedation. These marmosets were housed as same-sex pairs; one randomly selected individual from each pair was sampled. Marmosets were transported individually to an adjacent, temperaturecontrolled (24 °C) observation room. After recording began, a caretaker restrained the conscious marmoset using blue leather welder gloves (Liberty Glove Inc., City of Industry, CA) while a second caretaker inserted a 26-gauge needle (0.5-in Sterican Hypodermic Needle; B. Braun, Melsungen, Germany) percutaneously into the saphenous vein33 and collected a 200-ml blood sample for use in another study. No topical analgesics were applied prior to blood sampling. If a first attempt at blood sampling was not successful, a second attempt was made. After blood sampling was completed, we stopped the audio recording.
Audio recording and data analysis We recorded behaviors and audible vocalizations using a digital camcorder (Sony DCR-SR72E; Tokyo, Japan). The frequency threshold for audible vocalizations was set at 20 kHz. We analyzed the USVs using dedicated Sonotrack software as well as the opensource software programs mp3DirectCut (mpesch3 audio tools, Frankfurt, Germany), SYRINX (Higher Concept Software, Reading, UK) and Avisoft SASLab Lite (Avisoft Bioacoustics, Glienicke, Germany). For each experimental condition, we determined the total number of audible vocalizations, the vocalization rate and the presence of ultrasonic frequency components for each group. For the blood draw experiment and in the imitation predator experiment, we used the MannWhitney U test to determine the median and the 25% and 75% interquartile range (IQR) for USV production rates. We considered P values 20 kHz (ref. 1). Because the presence of higher harmonics in the vocalizations, which extend into the ultrasonic range, may be a consequence of more intense (energetic) calling, the impact of vocal intensity should be considered. Greater energy in the fundamental (‘audible’) component may make the ultrasonic components easier to detect. Unfortunately, we were unable to determine the energy in the fundamental components without USV and the fundamental components with USV. More research is needed for detailed analysis of spectral energy, fundamental and harmonic frequencies in the vocalizations of marmosets. We found that audible calls with ultrasonic components were produced by marmosets under normal conditions and also in response to restraint stress and confrontation with a perceived predator. Because we were not able to distinguish between the production of USVs in normal and stressful conditions, USV production cannot be used as a reliable indicator of stress in marmosets. Whether the presence of ultrasonic frequencies in the vocalizations of marmosets has any biological relevance remains a salient question. Common marmosets have a hearing range of up to 36 kHz (ref. 23), and since production of USVs is energyconsuming, it is likely that vocalizations made at these high frequencies are purposeful1. For example, USVs might be used to detect objects and obstacles, to measure distances or to locate prey. It might serve as a private channel of communication that cannot be intercepted by predators or prey. Additionally, the ultrasonic component might contain information about the sender (as in bird song), although we did not detect any individual USV signatures in this study. Although the current data do not clarify the function of USV production in marmosets, our observations that a limited range of audible vocalizations produced by marmosets contain ultrasonic components offer new opportunities to study the biological relevance of marmoset vocalizations. Acknowledgments We thank Herma van der Wiel for critical reading of the manuscript and Donna Devine and Thea de Koning for editing the manuscript. We also thank Henk van Westbroek for optimizing the graphs and figures. This study was funded in part by a grant from the EUPRIM-Net Project. COMPETING FINANCIAL INTERESTS The authors declare no competing financial interests. Received 3 January 2014; accepted 7 May 2014 Published online at http://www.labanimal.com/
LAB ANIMAL
1. Murrant, M.N. et al. Ultrasonic vocalizations emitted by flying squirrels. PLoS ONE 8, e73045 (2013). 2. Arch, V.S., Grafe, T.U., Gridi-Papp, M. & Narins, P.M. Pure ultrasonic communication in an endemic Bornean frog. PLoS ONE 4, e5413 (2009). 3. Arch, V.S. & Narins, P.M. “Silent” signals: Selective forces acting on ultrasonic communication systems in terrestrial vertebrates. Anim. Behav. 76, 1423–1428 (2008). 4. Brudzynski, S.M. Communication of adult rats by ultrasonic vocalization: biological, sociobiological, and neuroscience approaches. ILAR J. 50, 43–50 (2009). 5. Yu, P. et al. The effects of repeated early deprivation on ultrasonic vocalizations and ontogenetic development in mandarin vole pups. Behav. Processes 88, 162–167 (2011). 6. Clausen, K.T., Malmkvist, J. & Surlykke, A. Ultrasonic vocalizations of kits during maternal kit-retrieval in farmed mink, Mustela vison. Appl. Anim. Behav. Sci. 114, 582–592 (2008). 7. Bezerra, B.M. & Souta, A. Structure and usage of the vocal repertoire of Callithrix jacchus. Int. J. Primatol. 29, 671–701 (2008). 8. Epple, G. Comparative studies on vocalization in marmoset monkeys (Hapalidae). Folia Primatol. (Basel) 8, 1–40 (1968). 9. Goldman, J.A. Acoustic communication in the common marmoset monkey (Callithrix jacchus jacchus) measurements of transmissions and responses. PhD Dissertation, York University, Toronto, Canada (2000). 10. Jones, B.S. Vocal behaviour of the common marmoset: structure and function of selected calls. Primate Eye 53, 22–23 (1993). 11. Mendes, S.L., Vielliard, J.M.E. & De Marco, P. in The Smallest Anthropoids: The Marmoset/Callimico Radiation (Developments in Primatology: Progress and Prospects) (eds. Ford, S.M., Porter, L.M. & Davis, L.C.) 63–84 (Springer, New York, NY, 2009). 12. Miller, C.T., Mandel, K. & Wang, X. The communicative content of the common marmoset phee call during antiphonal calling. Am. J. Primatol. 72, 974–980 (2010). 13. Morrill, R.J., Thomas, A.W., Schiel, N., Souto, A. & Miller, C.T. The effect of habitat acoustics on common marmoset vocal signal transmission. Am. J. Primatol. 75, 904–916 (2013). 14. Norcross, J.L., Newman, J.D. & Cofrancesco, L.M. Context and sex differences exist in the acoustic structure of phee calls by newly-paired common marmosets (Callithrix jacchus). Am. J. Primatol. 49, 165–181 (1999). 15. Pistorio, A.L., Vintch, B. & Wang, X. Acoustic analysis of vocal development in a New World primate, the common marmoset (Callithrix jacchus). J. Acoust. Soc. Am. 120, 1655–1670 (2006). 16. Ramsier, M.A. et al. Primate communication in the pure ultrasound. Biol. Lett. 8, 508–511 (2012). 17. Stevenson, M.F. & Poole, T.B. An ethogram of the common marmoset (Calithrix jacchus jacchus): general behavioural repertoire. Anim. Behav. 24, 428–451 (1976). 18. Stevenson, M.F. & Rylands, A.B. in Ecology and Behavior of Neotropical Primates vol. 2. (eds. Mittermeier, R.A., Rylands, A.B., Coimbra-Filho, A.F. & Fonseca, G.A.B.) 131–222 (World Wildlife Fund, Washington, DC, 1988). 19. Watson, C.F. & Caldwell, C.A. Neighbor effects in marmosets: social contagion of agonism and affiliation in captive Callithrix jacchus. Am. J. Primatol. 72, 549–558 (2010). 20. Yamaguchi, C., Izumi, A. & Nakamura, K. Temporal rules in vocal exchanges of phees and trills in common marmosets (Callithrix jacchus). Am. J. Primatol. 71, 617–622 (2009). 21. Yamaguchi, C., Izumi, A. & Nakamura, K. Time course of vocal modulation during isolation in common marmosets (Callithrix jacchus). Am. J. Primatol. 72, 681–688 (2010). 22. Anderson, J.W. The production of ultrasonic sounds by laboratory rats and other mammals. Science 119, 808–809 (1954).
Volume 43, No. 9 | SEPTEMBER 2014 319
23. Osmanski, M.S. & Wang, X. Measurement of absolute auditory thresholds in the common marmoset (Callithrix jacchus). Hear. Res. 277, 127–133 (2011). 24. Gerber, P., Schnell, C.R. & Anzenberger, G. Behavioral and cardiophysiological responses of common marmosets (Callithrix jacchus) to social and environmental changes. Primates 43, 201–216 (2002). 25. Abbott, D.H., Barnett, D.K., Colman, R.J., Yamamoto, M.E. & Schultz-Darken, N.J. Aspects of common marmoset basic biology and life history important for biomedical research. Comp. Med. 53, 339–350 (2003). 26. Mansfield, K. Marmoset models commonly used in biomedical research. Comp. Med. 53, 383–392 (2003). 27. Burman, O.H.P., Ilyat, A., Jones, G. & Mendl, M. Ultrasonic vocalizations as indicators of welfare for laboratory rats (Rattus norvegicus). Appl. Anim. Behav. Sci. 104, 116–129 (2007). 28. Kikusui, T., Nishizawa, D., Takeuchi, Y. & Mori, Y. Conditioned fear-related ultrasonic vocalizations are emitted as an emotional response. J. Vet. Med. Sci. 65, 1299–1305 (2003). 29. Takahashi, N., Kashino, M. & Hironaka, N. Structure of rat ultrasonic vocalizations and its relevance to behavior. PLoS ONE 5, e14115 (2010). 30. Wöhr, M. & Schwarting, R.K. Ultrasonic communication in rats: can playback of 50-kHz calls induce approach behavior? PLoS ONE 2, e1365 (2007). 31. Portfors, C.V. Types and functions of ultrasonic vocalizations in laboratory rats and mice. J. Am. Assoc. Lab. Anim. Sci. 46, 28–34 (2007). 32. Williams, W.O., Riskin, D.K. & Mott, A.K. Ultrasonic sound as an indicator of acute pain in laboratory mice. J. Am. Assoc. Lab. Anim. Sci. 47, 8–10 (2008). 33. Jagessar, S.A. et al. Overview of models, methods, and reagents developed for translational autoimmunity research in the common marmoset (Callithrix jacchus). Exp. Anim. 62, 159–171 (2013).
34. Cagni, P., Sampaio, A.C., Ribeiro, N.B. & Barros, M. Immediate, but no delayed, behavioral response to a snake model by captive black tufted-ear marmosets. Behav. Processes 87, 241–245 (2011). 35. Clara, E., Tommasi, L. & Rogers, L.J. Social mobbing calls in common marmosets (Callithrix jacchus): effects of experience and associated cortisol levels. Anim. Cogn. 11, 349–358 (2008). 36. Emile, N. & Barros, M. Recognition of a 3D snake model and its 2D photographic image by captive black tufted-ear marmosets (Callithrix penicillata). Anim. Cogn. 12, 725–732 (2009). 37. Kaplan, G., Pines, M.K. & Rogers, L.J. Stress and stress reduction in common marmosets. Appl. Anim. Behav. Sci. 137, 175–182 (2012). 38. Norcross, J.L. & Newman, J.D. Context and gender-specific differences in the acoustic structure of common marmoset (Callithrix jacchus) phee calls. Am. J. Primatol. 30, 37–54 (1993). 39. Boere, V., Paludob, G.R., Pianta, G., Canale, C. & Tomaz, C. Effects of novelty, isolation stress, and environmental enrichment on some haematological parameters in marmosets (Callithrix penicillata). Comp. Biochem. Phys. Pharm. Tox. 119, 261–274 (2003). 40. Ely, A., Freer, A., Windle, C. & Ridley, R.M. Assessment of cage use by laboratory-bred common marmosets (Callithrix jacchus). Lab. Anim. 32, 427–433 (1998). 41. Searcy, Y.M. & Caine, N.G. Hawk calls elicit alarm and defensive reactions in captive Geoffroy’s marmosets (Callithrix geoffroyi). Folia Primatol. (Basel) 74, 115–125 (2003). 42. Winter, M. in Biology and Behaviour of Marmosets: Proceedings of the Marmoset Workshop, Göttingen, West Germany, 2–5 September 1977 (eds. Rothe, H., Wolters, H.J. & Hearn, J.P.) 127–139 (H. Rothe, Göttingen, Germany, 1978).
npg
© 2014 Nature America, Inc. All rights reserved.
Research Note
320 Volume 43, No. 9 | SEPTEMBER 2014
www.labanimal.com
