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Daily patterns of courtship and mating behavior in the male Japanese quail
223
Processes, 7 (1982) 223-233 Elsevier Scientific Publishing Company, Amsterdam - Printed in The Netherlands
Behavioural
DAILY PATTERNS OF COURTSHIP MALE JAPANESE QUAIL
AND MATING
BEHAVIOR
IN THE
MARY ANN OTTINGER, WOLFGANG M. SCHLEIDT*, and ESTELLE RUSSEK** Department
of Poultry
*Department
of Zoology,
**Department
(Accepted
Science,
University
University
of Dairy Science,
of Maryland,
of Maryland,
University
College
of Maryland,
College
Park, MD 20742
Park, MD 20742 College
(U.S.A.)
(U.S.A.)
Park, MD 20742
(U.S.A.)
11 February 1982)
ABSTRACT Ottinger, M.A., Schleidt, W.M. and Russek, E., 1982. Daily patterns of courtship and mating behavior in the male Japanese quail. Behav. Processes, 7: 223-233. Crowing behavior was monitored constantly in male Japanese quail housed singly over 30 successive days. The photoperiod was 16h of light and 8 h of dark. A daily pattern in crowing was observed in which the frequencies were elevated in the afternoon and at the beginning of darkness. However, peak crowing occurred 2 h prior to the onset of light. These rhythms were highly correlated among individuals and extremely repeatable over the sequential days of observation. In a second experiment, males which were paired with females were observed for frequencies of crowing, courtship, and mating behavior during the lighted portion of the day. In this experiment, the same photoperiod (16L:8D) was maintained. Paired males exhibited a daily pattern in crowing similar to that observed in the singly housed males. The frequency of mating was the highest between 1200 and 1300 h and lowest at 1400 h. Mating success was highest at midday, as were the number of males exhibiting mating behavior. These diurnal patterns in sexual behavior may depend on environmental cues such as photoperiod, which, in turn, may stimulate endocrine triggers.
INTRODUCTION
A number of rhythms have been reported for birds. In the domestic fowl, researchers have observed diurnal feeding patterns which differ with the reproductive state and the lighting schedule(Savory, 1980). Activity rhythms have, in some cases, been related to pineal rhythms (Takahashi and Menaker, 1979; Binkley, 1980; Deguichi, 1980), and a number of daily endocrine rhythms have been documented in birds (Boisson and Assenmacher, 1970; Meir, 1973; Noewcomer, 1974; Farner, 1980; Tanaka and Kamiyoski, 1980; Wada, 1980). There is evidence for a daily pattern in testosterone in the rooster and in the male Japanese quail (Schanbacher et al., 1974; Ottinger and Brinkley; 1979a). Moreover, semen volume in roosters has been documen-
0376-6357/82/0000-0000/802.75
0 1982 Elsevier Scientific Publishing Company
224
ted to vary diurnally, with highest volumes obtained in the afternoon (Lake and Wood-Gush, 1956). Many hormone-dependent behaviors, such as sexual behavior, have been reported to vary during the day. Both domestic ducks and wild mallards exhibited daily variations in social behavior and sexual behavior (Balthazart, 1976a, b). In the mallards, this was a bimodal activity pattern with increased activity in the morning and afternoon and highest frequencies of sexual behavior around noon (Balthazart, 1976b). Correlations were observed between variations in testosterone or FSH and sexual behavior and social displays (Balthazart and Hendrick, 1979). The domestic chicken exhibited a higher frequency of mating in the afternoon (Lake and Wood-Gush, 1956). Male Japanese quail in the field vocalized predominantly before sunrise and after dusk (Schwartz andSchwartz, 1949). In the laboratory, males vocalized predominantly in the early morning and late afternoon. Activity occurred in short period rhythms of 1 min and 2 h and was suppressed in paired males (Guyomarc’h and Thibout, 1969). Crowing in male quail has been found to de. pend on the presence of adequate circulating concentrations of testosterone or a metabolite of testosterone (Sachs, 1969; Adkins and Adler, 1972; Adkins and Pniewski, 1978; Ottinger and Brinkley, 1978a). Similarly, mating behavior was also dependent on adequate testosterone concentrations (Adkins and Adler, 1972; Balthazart et al., 1979; Ottinger and Brinkley, 1978a; Adkins et al., 1980). Since daily variations in social and sexual behavior have been observed in ducks and there is a relationship to endocrine changes (Balthazart, 1976a, b), it appears likely that similar patterns would occur for hormone-dependent behavior in other avian species. Daily patterns in crowing have been reported in Japanese quail (Schwartz and Schwartz, 1949; Guyomarc’h and Thibout, 1969). However, daily variation in mating behavior has not been examined. These experiments are designed to investigate daily patterns in courtship and mating behavior in the male Japanese quail. In experiment 1, males were individually caged in an anechoic chamber and crowing was recorded over 30 days. The objective of this experiment was to study variations in the frequency of crowing over successive days. In experiment II, courtship and mating behavior were observed in paired males during the lighted portion of the day. The objective this experiment was to study a) daily patterns in paired males, and b) differences in crowing frequencies in paired versus individually housed males. MATERIALS
Experimental
AND METHODS
animals
Male Japanese quail (Coturnix coturnix japonica) were hatched from eggs obtained from the University of Maryland random bred colony. At the time of the experiments, males were between 5 and 6 months of age. The photo-
225
period was 16 h of light and 8 h of dark with other environmental conditions remaining constant. All animals were provided feed (Purina Game Bird Feed) and water ad libitum.
Behavior measures The sexual behavior of the male Japanese quail and the methods of quantification have been described previously (Mather and Wilson, 1964; Adkins and Adler, 1972; Ottinger and Brinkley, 1979b). Crowing, a specific vocalization emitted by the male in breeding condition (Guyomarc’h and Thibout, 1969; Potash, 1972), was monitored in both experiments. In addition, strutting, which is a courtship behavior, mating attempts and completed matings (termed cloaca1 contacts) were recorded for each individual. Finally, a general record of activity was maintained for each 15 min period. Activity was assessed according to the following scale: (1) resting and sleeping, (2) feeding, sporadic movement, and some crowing, and (3) high activity including feeding, crowing, sexual activity, and constant movement.
Experiment
1
Three males were individually housed in 30.5 X 23 X 23 cm cages in an anechoic chamber. The three cages were placed at graded distances from, a microphone so that the recorded amplitude of crowing depended on the distance of the clling bird’s cage. The microphone was connected via a preamplifier to a Bruel and Kjaer level recorder. On the recording chart each crowing call was clearly separable, and distinct from other, incidental sounds. Crowing was monitored at all times for 30 consecutive days. Animals were fed and watered at varied times several times a week to avoid entrainment to feeding at a particular time of day. The photoperiod was 16L:8D with lights on at 0600 h and off at 2200 h. Records were quantified for each individual per hour. Calls of each male were first identified by observation and notation of the resulting sound pressure level on the recording chart. Since the distance of each male from the microphone differed from that of the other males, quantification of an individual’s vocalizations was possible with a minimum of error. Initially, visual observations were conducted for several hours on several days to determine the identity of the male with his vocalizations. Additional observations were made periodically verify measurements during the experiment, Statistical analysis for significant changes in frequency of crowing over time was done by a repeated measures analysis of variance (Winer, 1978) and Newman-Keuls multiple comparison test (Zar, 1974), using the error terms from the repeated measures analysis. Analysis by autocorrelation (Chatfield, 1980) was conducted to examine the occurrence of repeating patterns in crowing activity.
226
Experiment
II
Males were housed with females in battery cages, each measuring 18 X 18 X 20 cm. A total of 10 paired males were observed during the lighted portion of the day. The relative level of activity observed at night in a preliminary study was low, with a gradual increase 2-3 h prior to “lights on”. This coincides with activity rhythms observed in quail by Wada (1980). Therfore the observations were limited to lighted hours. All observations were conducted by one person, and arranged in 2 or 3 h blocks/day to avoid observer fatigue during a 2-week period of time. The oservation periods, which were assigned at random prior to the experiment, totalled 32 h of observations or two replicates of the 16 h lighted portion of the day. Behavior was recorded in 15 min segments. Males were monitored for general activity level, crowing, strutting, mating. attempts, and cloaca1 contacts. Analysis was done using a repeated measures analysis of variance. Sources of variance were computed as individuals, time, and error. A test for compound symmetry (Winer, 1978) was done to verify the necessary assumptions regarding correlations among observations recorded on the same individuals. An attempt at the cosinor method, as described by Tong (1976), failed because the overall fit to the multiple regression was rather poor. RESULTS
Experiment
I
The frequency of crowing varied in a repeating pattern every 24 h. The graph (Fig. 1) of crowing shows the average frequency/h as computed from NIGHT
DAY
I
DAY
NIGHT
DAY
3
2
I
NIGHT
DAYS
Fig. 1. Average
frequency
of crowing
per hour over 72 h.
DAY
NIGHT
221
MALES I. 2. -
LAG
2
-I
(Hours)
I
Fig. 2. Correlogram
representing
correlation
of crowing
activity
over 24 h.
the data for individuals over several successive days. There were brief, significant increases in crowing approximately 1 h prior to the onset of darkness. In addition, there were highly significant peaks (P < 0.01) 2 h before the onset of light. This pattern was extremely stable throughout the experiment and highly significant (P < 0.01) over a period of 24 h as is shown by the correlogram (Fig. 2). Comparison of individuals also showed highly correlated patterns of crowing (P < 0.01).
Experiment
II
Activity, as expressed on a scale of 1-3, varied during the day (Fig. 3). There were periods of high activity lasting an hour or longer in the early morning, midafternoon, and at the end of the day, The frequency of crowing/h (Fig. 4) was elevated at the start of the day and in the mid and late afternoon. There was a cessation and then sharp increase in crowing prior to the end of the day. When examined in 15 min periods (Fig. 4), crowing frequencies actually oscillated irregularly at a relatively low level in the morning and appeared to fluctuate at a higher level in the afternoon. There was a 2.25 h period to the end of the day in which almost no crowing occurred. In the 15 min prior to “lights off” there was a surge in crowing. Statistical analysis showed significant changes in crowing frequency over the time (P < O.Ol), as well as significant differences in crowing frequency among individuals (P < 0.01). The average number of crows over a day ranged from 2.2/h in one individual to 8.7 crows/h in another individual. Although these average frequencies differed, patterns of change were similar for all individuals and significant peaks (P < 0.05) in crowing occurred at 0700,1600,1800,1900
228
I = Preen,
Rest
2 = Moderate 3=
Htgh
Actlwty
Actlwty
HOUR
Fig. 3. Activity level during the lighted portion of the day as expressed on a scale of l-3. Bars represent averages for 15 min periods and the line represents change/h.
HOUR
Fig. 4. Frequency of crowing lighted portion of the day.
per hour (mean
? SEM)
and per 15 min periods
during the
and 2200 h. (A peak is defined as a significant increase followed by a significant decrease). The lowest frequency occurred at 2100 h. There were no significant correlations between the pattern in crowing and the other behavioral patterns in time. Mating behavior, particularly the number of mating attempts, was most frequent in the late morning and evening (Fig. 5). In terms of these daily patterns in behavior, there was elevated mating activity at 1000,1200,1300, 1500, 2000 through 2200 h, with significant (P < 0.05) peaks in the afternoon and evening. Lowest frequencies occurred at 1900 and 1400 h. The number of cloaca1 contacts followed a similar pattern with significant (P < 0.05) lows at 0900 and 1600 h. The average number of mating attempts over
229
I
0600
1000
1400
1800
2200
HOUR
Fig. 5. Mating behavior per hour (mean ? SEM). Unbroken tempts and dashed line represents cloaca1 contacts.
line represents
mating
at-
the day differed (F’ < 0.05) over the individuals and ranged from 2.5 to 4.56 attempts/h. Cloaca1 contacts averaged 1.06 to 2.13/h, depending on the individual. Again, behavioral patterns were parallel for individuals and individual differences corresponded to higher overall frequencies of behavior for one male as compared to another. There was a positive correlation within individuals (r = 0.69; P < 0.01) between mating attempts and cloaca1 contacts. DISCUSSION
Experiment
I
The 24 h pattern in crowing observed in singly housed males was in agreement, for the most part, with previous reports. There were peaks in crowing associated with the onset of light and dark phases of the photoperiod, similar to both laboratory and wild males (Schwartz and Schwartz, 1949; Guyomarc’h and Thibout, 1969). The expected high frequency of crowing from unpaired males (Potash, 1972) was confirmed in this experiment. However, the magnitude of change observed in these experiments was greater than previously reported, possibly due to differences in recording methods. In addition, the “predawn” peak occurred approximately 2 h before the onset of light, possible in anticipation of light rather than due to behavior associated with a slow increase in light as would occur with an actual dawn. It is interesting to note that this peak in crowing was extremely stable over the course of the experiment. Parenthetically, it should be noted that additional observations of crowing in the migratory quail show a similar pre-dawn peak; when the light is not switched on at the regular time, an excessively high level of crowing activity is maintained in darkness for several hours (Schleidt, unpublished data). In addition, the change in crowing activity prior to “lights off” was
230
relatively small in comparison to that which occurred prior to “lights on”. The 24 h pattern in crowing was repeated very regularly by all individuals. A likely environmental trigger for this rhythm is photoperiod, based on previous reports of the association of crowing with the onset of light and dark (Guyomarc’h and Thibout, 1969; Potash, 1972), as well as from observations in these experiments. There were no regular bouts of l-2 min or 2 h observed as reported by Guyomarc’h and Thibout (1969). Rather the fluctuations in crowing frequency were irregular and small, relative to the peak frequencies in crowing. It did appear that often the males tended to crow simultaneously. Experiment
II
The pattern of crowing observed in the paired males was similar to the rhythm in singly housed males during the day. This implies that when the behavior was viewed in 15 min periods rather than 1 h periods, it became apparent that the fluctuations in crowing were due to greater variation as well as higher rate of crowing. The decline in crowing between 2000 and 2100 h was actually due to a virtual cessation in crowing between 1915 and 2130 h. Moreover, the increase in crowing accompanying the end of the light phase concentrated activity, possibly in anticipation of the beginning of darkness. It is interesting to note that the number of crows in singly housed males was 30-50% higher than in paired males. It was observed by Guyomarc’h and Thibout (1969) that the amount of crowing by a male declined markedly when a female was temporarily housed with him. It appears from our data that long term pairing also has a depressing effect upon the frequency of vocalization in the male. However, in spite of the depression in crowing, the same pattern in crowing was maintained. Moreover, individual differences arose in the overall frequency rather than the daily pattern in crowing. This seemingly characteristic “level of crowing” in an individual might have a number of behavioral or physiological reasons, such as individual variation in hormone concentrations. Highest consistent activity in terms of movement, feeding , social interaction and dusting occurred in early morning, late afternoon and the hour prior to change in lighting. This early morning surge in activity is in agreement with observations by Wada (1980), who found diurnal activity patterns with increased activity at the onset of light. Mating attempts and cloaca1 contacts were frequent in the mid and late morning and in the evening, with the exception of 1400-1700 h. At those times there was a dramatic drop and then a brief rise in mating attempts, followed by another drop in mating attempts; whereas, the number of cloaca1 contacts appeared to drop uniformly. Analysis of these data according to the number of successful mating attempts showed that the greatest number of successful attempts occurred in the early afternoon. The number of animals exhibiting mating behavior also declined after the decline in mating success. This variation in mating behavior is somewhat similar to that observed in the
231
duck (Balthazart and Hendrick, 1979). Since the females were mostly afternoon egglayers, this decline may have been a response, in part, to the egglaying cycle and receptivity of the female. Since the behaviors observed are hormone dependent, it is possible that daily patterns in plasma steroid concentrations may relate to daily fluctuations in behavior. Testosterone concentrations peak at night in the male quail approximately 6 h prior to the observed peak in crowing (Ottinger and Brinkley, 1978b; Ottinger, 1979c). The timing of this peak is similar to the rooster (Schanbacher et al., 1974; Tanaka and Kamiyoski, 1980). There is also a daily nadir in plasma testosterone at approximately 1700 h with relatively higher con centrations in the morning and evening (Ottinger and Brinkley, 1979b). If the 24 h rhythm in crowing observed in this experiment is examined in light of the hormonal information, peak crowing occurred approximately 6 h after the peak in testosterone. However, since testosterone concentrations were examined at 2 or 3 h intervals, this lag time may still be somewhat of an approximation. There is not as clear a relationship between mating activity and plasma testosterone. However, there may be an association between minor peaks in plasma testosterone and mating activity. Further research is necessary to clarify this question. In addition, there is evidence for a relationship between testosterone and/or dihydrotestosterone and sexual behavior on a short term basis (Balthazart, 197613; Balthazart and Hendrick, 1979; Balthazart et al., 1979), as well as on a seasonal basis for plasma gonadotropins, gonadal activity and breeding (Mattocks et al., 1976; Campbell et al., 1978; Wingfield and Farner, 1978). In conclusion, environmental factors, such as photoperiod, may regulate endocrine rhythms which, in turn, may regulate, after an appropriate lag time, steroid-dependent behavioral activity to result in daily as well as circadian rhythms in reproductive behavior. ACKNOWLEDGEMENTS
The authors wish to acknowledge the use of the computer science facilities at the University of Maryland for providing computer time. The authors also thank Ms. Louise Palmer and Ms. Manju Masson for work on the graphs. Scientific Article No. A2993 Contribution No. 6055 of the Agricultural Experiment Station (Department of Poultry Science). Published in part in the Abstracts of the Poultry Science Conference, 1979.
REFERENCES Adkins, E.K. and Adler, N.I., 1972. Hormonal control of behavior in the J. Comp. Physiol. Psychol., 81( 1): 27-36. Adkins, E.K. and Pniewski, E.E., 1978. Control of reproductive behavior in male quail. J. Comp Physiol. Psychol., 92(6): 1169-1177. Adkins, E.K., Boop, J.J., Koutnik, D.L., Morris, J.B. and Pniewski, E.E., evidence that androgen aromatization is essential for the activation of male quail. Physiol. Behav., 24: 441-446.
Japanese
quail.
by sex steroids 1980. Further copulation in
232 Balthazart, J., 1976a. A correlation analysis of the daily distribution of displays and sexual behaviors in semi-wild mallards. Psychol. Belg., 16: l-26. Balthazart, J., 1976b). Daily variations of behavioural activities and of plasma testosterone levels in the domestic duck Anas platyrhynchos. J. Zool., 180: 155-173. Balthazart, J. and Hendrick, J.C., 1979. Relationships between the daily variations of social behavior and of plasma FSH, LH, and testosterone levels in the domestic duck Anas platyrhynchos L. Behav. Processes, 4: 107-128. Balthazart, J., Massa, R. and Negri-Cesi, P., 1979. Photoperiodic control of testosterone metabolism, plasma gonadotropins, cloaca1 gland growth, and reproductive behavior in the Japanese quail. Gen. Comp. Endocrinol., 39: 222-235. Binkley, S., 1980. The functions of the pineal gland. In: A. Epple and M.H. Stetson (Editors), Avian Endocrinology. Academic Press, New York, NY, pp. 53-74. Boisson, J. and Assenmacher, I., 1970. Circadian rhythms in adrenal cortical activity in the quail. J. Interdiscip. Cycle Res., l(3): 251-265. Campbell, R.R., Ashton, S.A. Follett, B.K. and Leatherland, J.F., 1978. Seasonal changes in plasma concentrations of LH in the lesser snow goose (Anser caerulescens caerulescens). Biol. Reprod., 18: 663-668. Chatfield, C., 1980. The Analysis of Time Series: An Introduction, 2nd Ed., Chapman & Hall, London. Deguichi, T., 1980. Circadian rhythm of pineal enzymes in culture In: Y. Tanabe, K. Tanaka and T. Ookawa (Editors), Biological Rhythms in Birds. Neural and Endocrine Aspects. Japanese Scientific Societies Press, Tokyo and Springer-Verlag, New York, NY, pp. 139-148. Farner, D.S., 1980. Endogenous periodic functions in the control of reproductive cycles, In: Y. Tanabe, K. Tanaka and T. Ookawa (Editors), Biological Rhythms in Birds. Neural and Endocrine Aspects. Japanese Scientific Societies Press, Tokyo and Springer-Verlag, New York, NY, pp. 123-138. Guyomarc’h, J.C. and Thibout, E., 1969. Rhythms et cycles dans l’emission du chant chez la caille japonaise (Coturnix coturnixjaponica). Rev. Comp. Anim., 3(3): 37-49. Lake, P.E. and Wood-Gush, D.G.N., 1956. Diurnal rhythms in semen yields and mating behavior in the domestic cock. Nature (London), 178: 853. Mather, F.B. and Wilson, W.O., 1964. Postnatal testicular development in Japanese quail. Poult. Sci., 43: 860-864. Mattocks, P.W., Jr., Farner, D.S., and Follett, B.K., 1976. The annual cycle in luteinizing hormone in the plasma of intact and castrated white-crowned sparrows, Zonotrichia Zencophrys gambelii. Gen. Comp. Endocrinol., 30: 156-161. Meir, A.H., 1973. Daily hormone rhythms in the white-throated sparrow. Am. Sci., 61: 184-187. Newcomer, W.S., 1974. Diurnal rhythms of thyroid function in chicks. Gen. Comp. Endocrinol., 24: 65-73. Ottinger, M.A. and Brinkley, H.J., 1978a. Testosterone and sex-related behavior and morphology: relationship during maturation and in adult Japanese quail. Horm. Behav. 11: 175-182. Ottinger, M.A. and Brinkley, H.J., 1978b. Diurnal rhythm in peripheral concentrations of testosterone in male quail. Am. Zool., 18(3): 603. Ottinger, M.A. and Brinkley, H.J., 1979a. The ontogeny of crowing and copulatory behavior in Japanese quail. (Coturnix coturnixjaponica). Behav. Processes, 4: 43-51. Ottinger, M.A. and Brinkley, H.J., 1979b. Testosterone and sex-related physical characteristics during the maturation of the male Japanese quail (Coturnix coturnix japonica). Biol. Reprod., 20: 905-909. Ottinger, M.A., 1979c. Daily rhythms in serum testosterone and sexual behavior in the male Japanese quail. Poult. Sci., 58(4): 1090.
233
Potash, L.M., 1972. A signal detection problem and possible solution in Japanese quail (Coturnix coturnix japonica). Anim. Behav., 20: 192-195. Sachs, B.D., 1969. Photoperiodic control of reproductive behavior and physiology of the male Japanese quail. (Coturnix coturnix juponica). Horm. Behav., 1, 7-24. Savory, C.J., 1980. Diurnal feeding patterns in domestic fowls: a review. Appl. Anim. Ethol., 6: 71-82. Schanbacher, B.D., Gomes, W.R. and VanDemark, N.L., 1974. Diurnal rhythm in serum testosterone levels and thymidine uptake by testes in the domestic fowl. J. Anim. Sci., 38(6) 1245-1248. Schwartz, C.W. and Schwartz, E.R., 1949. The Game Birds in Hawaii. Board of Commissioners of Agriculture and Forestry, Territory of Hawaii. Tanaka, K. and Kamiyoski, M., 1980. Rhythms of steroid hormone secretion in the domestic fowl. In: Y. Tanabe, K. Tanaka and T. Ookawa (Editors), Biological Rhythms in Birds. Neural and Endocrine Aspects. Japanese Scientific Press, Tokyo and SpringerVerlag, New York, NY, pp. 169-178. Takahashi, J.S. and Menaker, M., 1979. Physiology of avian circadian pacemakers. Fed. Proc. Fed. Am. Sot. Exp. Biol., 38: 2583-2588. Tong, Y.L., 1976. Parameter estimation in studying circadian rhythms. Biometrics, 32: 85-94. Wada, M., 1980. Nature of the circadian pacemaker of Japanese quail and its possible involvement in photoperiodic testicular growth. In: Y. Tanabe, K. Tanaka and T. Ookawa (Editors), Biological Rhythms in Birds. Neural and Endocrine Aspects. Japanese Scientific Societies Press, Tokyo and Springer-Verlag, New York, NY, pp. 157-168. Winer, B.J., 1971. Statistical Principles in Experimental Design, 2nd edition, McGraw-Hill, New York, NY. Wingfield, J.C. and Farner, D.S., 1978. The annual cycle of plasma in LH and steroid hormones in feral populations of the white-crowned sparrow, Zonatrichiu Zeucophrys gambelii. Biol. Reprod., 19: 1046-1056. Zar, J.H., 1974. Biostatistical Analysis. Prentice-Hall, Inc. Englewood Cliffs, NJ, pp. 151-155.
Processes, 7 (1982) 223-233 Elsevier Scientific Publishing Company, Amsterdam - Printed in The Netherlands
Behavioural
DAILY PATTERNS OF COURTSHIP MALE JAPANESE QUAIL
AND MATING
BEHAVIOR
IN THE
MARY ANN OTTINGER, WOLFGANG M. SCHLEIDT*, and ESTELLE RUSSEK** Department
of Poultry
*Department
of Zoology,
**Department
(Accepted
Science,
University
University
of Dairy Science,
of Maryland,
of Maryland,
University
College
of Maryland,
College
Park, MD 20742
Park, MD 20742 College
(U.S.A.)
(U.S.A.)
Park, MD 20742
(U.S.A.)
11 February 1982)
ABSTRACT Ottinger, M.A., Schleidt, W.M. and Russek, E., 1982. Daily patterns of courtship and mating behavior in the male Japanese quail. Behav. Processes, 7: 223-233. Crowing behavior was monitored constantly in male Japanese quail housed singly over 30 successive days. The photoperiod was 16h of light and 8 h of dark. A daily pattern in crowing was observed in which the frequencies were elevated in the afternoon and at the beginning of darkness. However, peak crowing occurred 2 h prior to the onset of light. These rhythms were highly correlated among individuals and extremely repeatable over the sequential days of observation. In a second experiment, males which were paired with females were observed for frequencies of crowing, courtship, and mating behavior during the lighted portion of the day. In this experiment, the same photoperiod (16L:8D) was maintained. Paired males exhibited a daily pattern in crowing similar to that observed in the singly housed males. The frequency of mating was the highest between 1200 and 1300 h and lowest at 1400 h. Mating success was highest at midday, as were the number of males exhibiting mating behavior. These diurnal patterns in sexual behavior may depend on environmental cues such as photoperiod, which, in turn, may stimulate endocrine triggers.
INTRODUCTION
A number of rhythms have been reported for birds. In the domestic fowl, researchers have observed diurnal feeding patterns which differ with the reproductive state and the lighting schedule(Savory, 1980). Activity rhythms have, in some cases, been related to pineal rhythms (Takahashi and Menaker, 1979; Binkley, 1980; Deguichi, 1980), and a number of daily endocrine rhythms have been documented in birds (Boisson and Assenmacher, 1970; Meir, 1973; Noewcomer, 1974; Farner, 1980; Tanaka and Kamiyoski, 1980; Wada, 1980). There is evidence for a daily pattern in testosterone in the rooster and in the male Japanese quail (Schanbacher et al., 1974; Ottinger and Brinkley; 1979a). Moreover, semen volume in roosters has been documen-
0376-6357/82/0000-0000/802.75
0 1982 Elsevier Scientific Publishing Company
224
ted to vary diurnally, with highest volumes obtained in the afternoon (Lake and Wood-Gush, 1956). Many hormone-dependent behaviors, such as sexual behavior, have been reported to vary during the day. Both domestic ducks and wild mallards exhibited daily variations in social behavior and sexual behavior (Balthazart, 1976a, b). In the mallards, this was a bimodal activity pattern with increased activity in the morning and afternoon and highest frequencies of sexual behavior around noon (Balthazart, 1976b). Correlations were observed between variations in testosterone or FSH and sexual behavior and social displays (Balthazart and Hendrick, 1979). The domestic chicken exhibited a higher frequency of mating in the afternoon (Lake and Wood-Gush, 1956). Male Japanese quail in the field vocalized predominantly before sunrise and after dusk (Schwartz andSchwartz, 1949). In the laboratory, males vocalized predominantly in the early morning and late afternoon. Activity occurred in short period rhythms of 1 min and 2 h and was suppressed in paired males (Guyomarc’h and Thibout, 1969). Crowing in male quail has been found to de. pend on the presence of adequate circulating concentrations of testosterone or a metabolite of testosterone (Sachs, 1969; Adkins and Adler, 1972; Adkins and Pniewski, 1978; Ottinger and Brinkley, 1978a). Similarly, mating behavior was also dependent on adequate testosterone concentrations (Adkins and Adler, 1972; Balthazart et al., 1979; Ottinger and Brinkley, 1978a; Adkins et al., 1980). Since daily variations in social and sexual behavior have been observed in ducks and there is a relationship to endocrine changes (Balthazart, 1976a, b), it appears likely that similar patterns would occur for hormone-dependent behavior in other avian species. Daily patterns in crowing have been reported in Japanese quail (Schwartz and Schwartz, 1949; Guyomarc’h and Thibout, 1969). However, daily variation in mating behavior has not been examined. These experiments are designed to investigate daily patterns in courtship and mating behavior in the male Japanese quail. In experiment 1, males were individually caged in an anechoic chamber and crowing was recorded over 30 days. The objective of this experiment was to study variations in the frequency of crowing over successive days. In experiment II, courtship and mating behavior were observed in paired males during the lighted portion of the day. The objective this experiment was to study a) daily patterns in paired males, and b) differences in crowing frequencies in paired versus individually housed males. MATERIALS
Experimental
AND METHODS
animals
Male Japanese quail (Coturnix coturnix japonica) were hatched from eggs obtained from the University of Maryland random bred colony. At the time of the experiments, males were between 5 and 6 months of age. The photo-
225
period was 16 h of light and 8 h of dark with other environmental conditions remaining constant. All animals were provided feed (Purina Game Bird Feed) and water ad libitum.
Behavior measures The sexual behavior of the male Japanese quail and the methods of quantification have been described previously (Mather and Wilson, 1964; Adkins and Adler, 1972; Ottinger and Brinkley, 1979b). Crowing, a specific vocalization emitted by the male in breeding condition (Guyomarc’h and Thibout, 1969; Potash, 1972), was monitored in both experiments. In addition, strutting, which is a courtship behavior, mating attempts and completed matings (termed cloaca1 contacts) were recorded for each individual. Finally, a general record of activity was maintained for each 15 min period. Activity was assessed according to the following scale: (1) resting and sleeping, (2) feeding, sporadic movement, and some crowing, and (3) high activity including feeding, crowing, sexual activity, and constant movement.
Experiment
1
Three males were individually housed in 30.5 X 23 X 23 cm cages in an anechoic chamber. The three cages were placed at graded distances from, a microphone so that the recorded amplitude of crowing depended on the distance of the clling bird’s cage. The microphone was connected via a preamplifier to a Bruel and Kjaer level recorder. On the recording chart each crowing call was clearly separable, and distinct from other, incidental sounds. Crowing was monitored at all times for 30 consecutive days. Animals were fed and watered at varied times several times a week to avoid entrainment to feeding at a particular time of day. The photoperiod was 16L:8D with lights on at 0600 h and off at 2200 h. Records were quantified for each individual per hour. Calls of each male were first identified by observation and notation of the resulting sound pressure level on the recording chart. Since the distance of each male from the microphone differed from that of the other males, quantification of an individual’s vocalizations was possible with a minimum of error. Initially, visual observations were conducted for several hours on several days to determine the identity of the male with his vocalizations. Additional observations were made periodically verify measurements during the experiment, Statistical analysis for significant changes in frequency of crowing over time was done by a repeated measures analysis of variance (Winer, 1978) and Newman-Keuls multiple comparison test (Zar, 1974), using the error terms from the repeated measures analysis. Analysis by autocorrelation (Chatfield, 1980) was conducted to examine the occurrence of repeating patterns in crowing activity.
226
Experiment
II
Males were housed with females in battery cages, each measuring 18 X 18 X 20 cm. A total of 10 paired males were observed during the lighted portion of the day. The relative level of activity observed at night in a preliminary study was low, with a gradual increase 2-3 h prior to “lights on”. This coincides with activity rhythms observed in quail by Wada (1980). Therfore the observations were limited to lighted hours. All observations were conducted by one person, and arranged in 2 or 3 h blocks/day to avoid observer fatigue during a 2-week period of time. The oservation periods, which were assigned at random prior to the experiment, totalled 32 h of observations or two replicates of the 16 h lighted portion of the day. Behavior was recorded in 15 min segments. Males were monitored for general activity level, crowing, strutting, mating. attempts, and cloaca1 contacts. Analysis was done using a repeated measures analysis of variance. Sources of variance were computed as individuals, time, and error. A test for compound symmetry (Winer, 1978) was done to verify the necessary assumptions regarding correlations among observations recorded on the same individuals. An attempt at the cosinor method, as described by Tong (1976), failed because the overall fit to the multiple regression was rather poor. RESULTS
Experiment
I
The frequency of crowing varied in a repeating pattern every 24 h. The graph (Fig. 1) of crowing shows the average frequency/h as computed from NIGHT
DAY
I
DAY
NIGHT
DAY
3
2
I
NIGHT
DAYS
Fig. 1. Average
frequency
of crowing
per hour over 72 h.
DAY
NIGHT
221
MALES I. 2. -
LAG
2
-I
(Hours)
I
Fig. 2. Correlogram
representing
correlation
of crowing
activity
over 24 h.
the data for individuals over several successive days. There were brief, significant increases in crowing approximately 1 h prior to the onset of darkness. In addition, there were highly significant peaks (P < 0.01) 2 h before the onset of light. This pattern was extremely stable throughout the experiment and highly significant (P < 0.01) over a period of 24 h as is shown by the correlogram (Fig. 2). Comparison of individuals also showed highly correlated patterns of crowing (P < 0.01).
Experiment
II
Activity, as expressed on a scale of 1-3, varied during the day (Fig. 3). There were periods of high activity lasting an hour or longer in the early morning, midafternoon, and at the end of the day, The frequency of crowing/h (Fig. 4) was elevated at the start of the day and in the mid and late afternoon. There was a cessation and then sharp increase in crowing prior to the end of the day. When examined in 15 min periods (Fig. 4), crowing frequencies actually oscillated irregularly at a relatively low level in the morning and appeared to fluctuate at a higher level in the afternoon. There was a 2.25 h period to the end of the day in which almost no crowing occurred. In the 15 min prior to “lights off” there was a surge in crowing. Statistical analysis showed significant changes in crowing frequency over the time (P < O.Ol), as well as significant differences in crowing frequency among individuals (P < 0.01). The average number of crows over a day ranged from 2.2/h in one individual to 8.7 crows/h in another individual. Although these average frequencies differed, patterns of change were similar for all individuals and significant peaks (P < 0.05) in crowing occurred at 0700,1600,1800,1900
228
I = Preen,
Rest
2 = Moderate 3=
Htgh
Actlwty
Actlwty
HOUR
Fig. 3. Activity level during the lighted portion of the day as expressed on a scale of l-3. Bars represent averages for 15 min periods and the line represents change/h.
HOUR
Fig. 4. Frequency of crowing lighted portion of the day.
per hour (mean
? SEM)
and per 15 min periods
during the
and 2200 h. (A peak is defined as a significant increase followed by a significant decrease). The lowest frequency occurred at 2100 h. There were no significant correlations between the pattern in crowing and the other behavioral patterns in time. Mating behavior, particularly the number of mating attempts, was most frequent in the late morning and evening (Fig. 5). In terms of these daily patterns in behavior, there was elevated mating activity at 1000,1200,1300, 1500, 2000 through 2200 h, with significant (P < 0.05) peaks in the afternoon and evening. Lowest frequencies occurred at 1900 and 1400 h. The number of cloaca1 contacts followed a similar pattern with significant (P < 0.05) lows at 0900 and 1600 h. The average number of mating attempts over
229
I
0600
1000
1400
1800
2200
HOUR
Fig. 5. Mating behavior per hour (mean ? SEM). Unbroken tempts and dashed line represents cloaca1 contacts.
line represents
mating
at-
the day differed (F’ < 0.05) over the individuals and ranged from 2.5 to 4.56 attempts/h. Cloaca1 contacts averaged 1.06 to 2.13/h, depending on the individual. Again, behavioral patterns were parallel for individuals and individual differences corresponded to higher overall frequencies of behavior for one male as compared to another. There was a positive correlation within individuals (r = 0.69; P < 0.01) between mating attempts and cloaca1 contacts. DISCUSSION
Experiment
I
The 24 h pattern in crowing observed in singly housed males was in agreement, for the most part, with previous reports. There were peaks in crowing associated with the onset of light and dark phases of the photoperiod, similar to both laboratory and wild males (Schwartz and Schwartz, 1949; Guyomarc’h and Thibout, 1969). The expected high frequency of crowing from unpaired males (Potash, 1972) was confirmed in this experiment. However, the magnitude of change observed in these experiments was greater than previously reported, possibly due to differences in recording methods. In addition, the “predawn” peak occurred approximately 2 h before the onset of light, possible in anticipation of light rather than due to behavior associated with a slow increase in light as would occur with an actual dawn. It is interesting to note that this peak in crowing was extremely stable over the course of the experiment. Parenthetically, it should be noted that additional observations of crowing in the migratory quail show a similar pre-dawn peak; when the light is not switched on at the regular time, an excessively high level of crowing activity is maintained in darkness for several hours (Schleidt, unpublished data). In addition, the change in crowing activity prior to “lights off” was
230
relatively small in comparison to that which occurred prior to “lights on”. The 24 h pattern in crowing was repeated very regularly by all individuals. A likely environmental trigger for this rhythm is photoperiod, based on previous reports of the association of crowing with the onset of light and dark (Guyomarc’h and Thibout, 1969; Potash, 1972), as well as from observations in these experiments. There were no regular bouts of l-2 min or 2 h observed as reported by Guyomarc’h and Thibout (1969). Rather the fluctuations in crowing frequency were irregular and small, relative to the peak frequencies in crowing. It did appear that often the males tended to crow simultaneously. Experiment
II
The pattern of crowing observed in the paired males was similar to the rhythm in singly housed males during the day. This implies that when the behavior was viewed in 15 min periods rather than 1 h periods, it became apparent that the fluctuations in crowing were due to greater variation as well as higher rate of crowing. The decline in crowing between 2000 and 2100 h was actually due to a virtual cessation in crowing between 1915 and 2130 h. Moreover, the increase in crowing accompanying the end of the light phase concentrated activity, possibly in anticipation of the beginning of darkness. It is interesting to note that the number of crows in singly housed males was 30-50% higher than in paired males. It was observed by Guyomarc’h and Thibout (1969) that the amount of crowing by a male declined markedly when a female was temporarily housed with him. It appears from our data that long term pairing also has a depressing effect upon the frequency of vocalization in the male. However, in spite of the depression in crowing, the same pattern in crowing was maintained. Moreover, individual differences arose in the overall frequency rather than the daily pattern in crowing. This seemingly characteristic “level of crowing” in an individual might have a number of behavioral or physiological reasons, such as individual variation in hormone concentrations. Highest consistent activity in terms of movement, feeding , social interaction and dusting occurred in early morning, late afternoon and the hour prior to change in lighting. This early morning surge in activity is in agreement with observations by Wada (1980), who found diurnal activity patterns with increased activity at the onset of light. Mating attempts and cloaca1 contacts were frequent in the mid and late morning and in the evening, with the exception of 1400-1700 h. At those times there was a dramatic drop and then a brief rise in mating attempts, followed by another drop in mating attempts; whereas, the number of cloaca1 contacts appeared to drop uniformly. Analysis of these data according to the number of successful mating attempts showed that the greatest number of successful attempts occurred in the early afternoon. The number of animals exhibiting mating behavior also declined after the decline in mating success. This variation in mating behavior is somewhat similar to that observed in the
231
duck (Balthazart and Hendrick, 1979). Since the females were mostly afternoon egglayers, this decline may have been a response, in part, to the egglaying cycle and receptivity of the female. Since the behaviors observed are hormone dependent, it is possible that daily patterns in plasma steroid concentrations may relate to daily fluctuations in behavior. Testosterone concentrations peak at night in the male quail approximately 6 h prior to the observed peak in crowing (Ottinger and Brinkley, 1978b; Ottinger, 1979c). The timing of this peak is similar to the rooster (Schanbacher et al., 1974; Tanaka and Kamiyoski, 1980). There is also a daily nadir in plasma testosterone at approximately 1700 h with relatively higher con centrations in the morning and evening (Ottinger and Brinkley, 1979b). If the 24 h rhythm in crowing observed in this experiment is examined in light of the hormonal information, peak crowing occurred approximately 6 h after the peak in testosterone. However, since testosterone concentrations were examined at 2 or 3 h intervals, this lag time may still be somewhat of an approximation. There is not as clear a relationship between mating activity and plasma testosterone. However, there may be an association between minor peaks in plasma testosterone and mating activity. Further research is necessary to clarify this question. In addition, there is evidence for a relationship between testosterone and/or dihydrotestosterone and sexual behavior on a short term basis (Balthazart, 197613; Balthazart and Hendrick, 1979; Balthazart et al., 1979), as well as on a seasonal basis for plasma gonadotropins, gonadal activity and breeding (Mattocks et al., 1976; Campbell et al., 1978; Wingfield and Farner, 1978). In conclusion, environmental factors, such as photoperiod, may regulate endocrine rhythms which, in turn, may regulate, after an appropriate lag time, steroid-dependent behavioral activity to result in daily as well as circadian rhythms in reproductive behavior. ACKNOWLEDGEMENTS
The authors wish to acknowledge the use of the computer science facilities at the University of Maryland for providing computer time. The authors also thank Ms. Louise Palmer and Ms. Manju Masson for work on the graphs. Scientific Article No. A2993 Contribution No. 6055 of the Agricultural Experiment Station (Department of Poultry Science). Published in part in the Abstracts of the Poultry Science Conference, 1979.
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