-The present study was carried out on a Palearctic-Indian migratory species, the blackheaded bunting (Emberiza melanocephala), to understand the importance of photoperiodism and circannual rhythms in determining seasonality in changes in body mass and testis size in birds. An initial experiment determined the effects of duration and intensity of light on photoperiodic induction. The birds were exposed to different photoperiods (hours of light:hours of darkness; 11.5L:12.5D, 12L:12D, 12.5L:11.5D and 13L:11D) at the same (~ 450 lux) light intensity, and to 13L:11D at different light intensities (50-, 100-, 400-, 800-and 1000-lux). The induction and subsequent regression of photoperiodic responses were dependent upon duration and intensity of the light period until these reached threshold. A second experiment investigated if an endogenous seasonal rhythm underlies photoperiodism in buntings. Birds maintained since February on a 8L:16D photoperiod (a non-inductive short day length invariably used to ensure photosensitivity in photoperiodic species) were subjected periodically to 16L:8D (a long day length), one group every month from mid-March to mid-August. The magnitude of long day response in body mass and testes decreased as the duration of the short days progressed, but testicular response was restored in birds that were exposed to long days in July and August. The birds exposed simultaneously to short, long, and natural day lengths for 32 weeks underwent an induction-regression cycle under long days and natural day lengths, but not under short days in which a decrease in body mass occurred after about 20 weeks. The last experiment examined the importance of latitudinal migration on photoperiodism, by comparing the response to long days of three groups which included birds from populations those were held in the outdoor aviary for 1 or 2 years at 27° N and those immediately arrived from their breeding grounds (~ 40° N). There was no difference in the photoperiodic induction among the three groups, indicating that neither experience to changing photoperiods during a migratory journey, nor to long photoperiods at breeding grounds, were critical for a subsequent response (initiationtermination-reinitiation) cycle. Taken together, these findings suggest that (1) the blackheaded bunting has its own endogenous timing program, which is regulated by the photoperiod, and (2) the photoperiodic programs of bunting are flexible enough to accommodate variations in the amplitude of environmental cycles. Thus, it appears that photoperiodism has evolved independently of the evolution of migration in this species.bunting / intensity / photoperiod / photoperiodic response system / photosensitivity / seasonality

This study analyzed the role of day length in regulation of seasonal body fattening and testicular growth in a latitudinal Palaearctic-Indian migrant, the redheaded bunting (Emberiza bruniceps). When exposed to increasing photoperiods (hours of light: hours of darkness; 11.5L:12.5D, 12L:12D, 12.5L:11.5D, 13L:11D, 14L:10D, and 18L:6D) for 9-12 weeks, buntings responded in a photoperiod-dependent manner and underwent growth and regression cycle under photoperiods of > or =12 hr per day. Also, the response to a long photoperiod of birds that were held under natural photoperiods at 27 degrees N for 2 years was similar to those who arrived the same year from their breeding grounds ( approximately 40 degrees N), suggesting that the experience of higher amplitude day-night (light-dark, LD) cycles during migratory and breeding seasons were not critical for the subsequent response (initiation-termination-reinitiation) cycle. Another experiment examined entrainment of the circadian photoperiodic rhythm in buntings by subjecting them to T=24+/-2 hr LD-cycles with 8 hr photophase and to T=22 and 24 hr with 11 hr photophase. The results showed a reduction in critical day length under T=22 hr LD-cycle. In the last experiment, we constructed an action spectrum for photoperiodic induction by exposing birds for 4.5 weeks to 13L:11D of white (control), blue (450 nm), or red (640 nm) light at irradiances ranging from 0.028 to 1.4 W m(-2). The threshold light irradiance for photoinduction was about 10-fold higher for blue light, than for red and white lights. These results conclude that the daily light of the environment regulates the endogenous program that times seasonal responses in body fattening and testicular cycles of the redheaded bunting.

In the present study, we asked the question whether physiological responses to day length of migratory redheaded bunting (Emberiza bruniceps) and nonmigratory Indian weaver bird (Ploceus philippinus) are mediated by the daily rhythm of melatonin. Melatonin was given either by injection at certain times of the day or as an implant. In series I experiments on the redheaded bunting, melatonin was administered by subcutaneous injections daily at zeitgeber time (ZT) 4 (morning) or ZT10 (evening) and by silastic capsules in photosensitive unstimulated buntings that were held in natural day lengths (NDL) at 27 degrees N beginning from mid February, and in artificial day lengths (ADL, 12L:12D and 14L:10D). Melatonin did not affect the photoperiod-induced cycles of gain and loss in body mass and testicular growth-involution, but there was an effect on temporal phasing of the growth-involution cycle of testes in some groups. For example, the rate of testicular growth and development was faster in birds that received melatonin injection at ZT4 in NDL, and was slower in birds that carried melatonin implants both in NDL and ADL. In series II experiments on Indian weaver birds, melatonin was given in silastic capsules in the first week of September when they still had large gonads. Birds were exposed for 12 weeks to short day length (8L:16D; group 1), to long day length (eight weeks of 16L:8D and four weeks of 18L:6D; group 2), or to both short and long day lengths (four weeks each of 8L:16D, 16L:8D, and 18L:6D; groups 3 and 4). Whereas groups 1 to 3 carried melatonin or empty implant from the beginning, group 4 received one after four weeks. All birds underwent testicular regression during the first four weeks irrespective of the photoperiod they were exposed to or the implant they carried in, and there was a slight re-initiation of testis growth in some birds during the next eight weeks of long day lengths. However, with the exception of group 2, there was no difference in mean testis volume during the period of experiment between the melatonin- and empty-implant birds. The data on androgen-dependent beak color also supported the observations on testes. Together, these results do not support the idea that the daily rhythm of melatonin is involved in the photoperiodic time measurement in birds. However, there may still be a role of melatonin in temporal phasing of the annual reproductive cycle in birds.

SUMMARY The effects of the duration and time of food availability on stimulation of the photoperiodic responses (fattening and gain in body mass, and growth and development of testes) were investigated in the migratory blackheaded bunting(Emberiza melanocephala). Two experiments were performed. Experiment 1 examined the effects of a reduction in the duration of food supply in buntings that were subjected to long day lengths (16 h:8 h L:D) and received food ad libitum (group I) or for restricted durations, coinciding with the end of the lights-on period, of 8 h (group II) and 4 h (group III). Buntings of group I gained in body mass, whereas there was a mixed response in group II (half the birds gained and half lost body mass), and all birds of group III lost body mass. There was no effect on testis growth in groups I and II, but testes grew more slowly in group III. Experiment 2 investigated the effects of both the duration and the time of food availability. Of five groups of birds, group I was exposed to an 8 h:16 h L:D photoperiod, and groups II-V were exposed to 16 h:8 h L:D. Whereas birds of groups I and II received food ad libitum, those of groups III-V were fed only for 5 h, at zt 0-5(group III), zt 5.5-10.5 (group IV) or zt 11-16 (group V), where zt =zeitgeber time and zt 0 refers to the beginning of the lights-on period. Apart from duration, the timing of food availability also had an effect on photoperiodic stimulation under the 16 h:8 h L:D photoperiod. Birds that were fed ad libitum fattened and gained in body mass, whereas among restricted feeding groups, only birds in the group fed during the first 5 h(zt 0-5, group III) showed a significant increase in body mass (albeit considerably lower than in the ad libitum group). Birds fed during the middle 5 h (zt 5.5-10.5, group IV) showed an intermediate response, and those fed during the last 5 h (zt 11-16, group V) lost body mass. Testicular growth was suppressed in birds that were fed for 5 h in the evening, but not in those fed for the same period in the morning or in the middle of the long day. Taken together, these results show that the duration of food supply and/or the time of day at which food is available affect photoperiodic stimulation of fattening and gain in body mass as well as the growth and development of gonads in the blackheaded bunting.

The properties of the circadian photoperiodic oscillator have been investigated in detail only in the Japanese quail. While the study of the quail is clearly very important, one cannot simply assume that other species, especially passerines that seem to have a different circadian organization than quail, function the same way. The current set of experiments was conducted to understand the entrainment and photoinduction of the circadian photoperiodic oscillator in a passerine species, the blackheaded bunting (Emberiza melanocephala). The experimental paradigm used skeleton photoperiods with two light periods, the first called the "entraining light pulse" (E-pulse) and the second called the "inducing light pulse" (I-pulse). Three experiments were performed on photosensitive male birds (N = 6-8/group). Experiment 1 investigated the effects of the temporal relationship between E- and I-pulses on photoperiodic induction. Buntings entrained to 8h: 16h L:D for 4 wk were released into constant dim light (LLdim, approximately 1 lux). Beginning on subjective day 8, they received for 8 wk, E- and I-pulses only at alternate cycles. While I-pulse was 1h and always began at zt 11.5, E-pulse varied in duration and timing (the 1h E-pulse beginning either at zt 0, zt 5, or zt 9, the 4h one beginning at zt 0 or zt 6, and the 10 h one at zt 0; zeitgeber time 0 = time of lights-on under 8h: 16h L:D prior to release into LLdim). A photoperiodic response was induced only when the E-pulse began at zt 0, and thus the beginning of E- and 1-pulses were separated by 11.5h. Experiment 2 determined whether the duration of the E-pulse influences the position of the photoinducible phase (phii) of the circadian photoperiodic oscillator. Birds were entrained to 1h:23h L:D or 10h: 14h L:D for 2 wk, and then exposed to 1h I-pulse at zt 11.5, zt 15, or zt 18.5 for another 8 wk. Photoperiodic induction occurred at all 3 zts in birds entrained to 10h but only at zt 11.5 in birds entrained to 1h, which infers the circadian rhythm of photoinducibility (CRP) in buntings was re-entrained when I-pulse fell at zt 15 and after. The last experiment examined the possibility of the re-entrainment of the CRP to light pulses falling at zt 15 and after. Birds received 1h I-pulse for 8wk at zt 15 following 2wk of 2.5h:21.5h L:D or 3.5h:20.5h L:D, or at zt 21.5 or zt 22.5 following 2 wk of 10h: 14h LD. Photoperiodic induction was consistent with the hypothesis of the re-entrainment of the CRP under these light-dark cycles. The I-pulse appeared to be interpreted as a "new dawn", and so the photoperiodic induction was determined by the coincidence of phii with the E-pulse. These results suggest a phase-dependent action of light on the circadian oscillator regulating photoperiodic responses in the blackheaded bunting. This could be a useful strategy for a photoperiodic species to regulate its seasonal responses in nature.