Response of restraint stress-selected lines of Japanese quail to heat stress and Escherichia coli challenge.

Japanese quail selected for divergent corticosterone response to restraint stress were evaluated for their susceptibility to heat stress and challenge with Escherichia coli. These quail lines are designated as high stress (HS), low stress (LS), and the random-bred control (CS) lines. Heat stress (35°C, 8 h/d) began at 24 d until the end of the study at 39 d. Birds were challenged with an aerosol spray containing 2 × 10(9) cfu of E. coli at 25 and 32 d. At 38 d, the surviving birds were necropsied and the intestinal tract was screened for both Salmonella and Campylobacter. Body weights of the CS birds were higher than both HS and LS at 17, 25, and 32 d. At 32 d, there was no difference in mortality between males and females and the CS line had higher mortality compared with the LS line with the HS line being intermediate. At 38 d, females of the CS line that were both heat stressed and challenged had a mortality incidence of 25%, which was significantly higher than male birds of the same line and treatment (5.3%). There was an increased incidence in Salmonella enterica serotype Agona isolation after heat stress, with the LS birds having less isolation than the HS birds. Mean corticosterone levels of male birds were not significantly affected by line, heat stress, or E. coli challenge; however, the LS line subjected to heat stress had one-third the level of the HS line, a difference identical to that seen in the original selection for response to restraint stress.

Fecal corticosterone metabolites and plasma corticosterone in Japanese quail selected for low or high plasma corticosterone responses to brief restraint.

Fecal corticosterone metabolites and plasma corticosterone in Japanese quail selected for low- or high-plasma corticosterone responses to brief mechanical restraint (low- and high-stress lines), and in a line of unselected quail, were measured in this study. No line differences were observed in baseline plasma corticosterone concentrations, but fecal corticosterone metabolite concentrations and daily fecal corticosterone metabolite production were 20% higher in quail of the high-stress line than in unselected or low-stress quail for males and females living together in group cages (P < 0.05). No differences were observed between lines in corticosterone metabolite concentrations and production for male birds in individual cages. Baseline plasma corticosterone concentrations, fecal corticosterone metabolite concentrations, and production appeared to be higher for males and females in group cages compared with males in individual cages. This difference might have been because of greater corticosterone secretion by male quail living in mixed sex groups than living individually. Correlations between baseline plasma corticosterone concentrations and fecal corticosterone metabolite concentrations in low-stress and high-stress quail, and for all birds combined, were r = 0.521 (P = 0.038), r = 0.604 (P = 0.013), and r = 0.431 (P = 0.002), respectively. The low- and high-stress lines that have been selected for low- and high-corticosterone responses differ in other characteristics, including growth and reproductive performance, and the current results are consistent with the assumption that these other differences are a consequence of greater daily corticosterone secretion in quail of the high-stress line.

Testosterone manipulation postcastration does not alter cloacal gland growth differences in male quail selected for divergent plasma corticosterone stress response.

Japanese quail selected for reduced (low-stress, LS) rather than exaggerated (high-stress, HS) plasma corticosterone response to brief restraint have consistently shown greater cloacal gland (CG) development, an androgen-dependent trait. In this study, the effects of testosterone implants on levels of plasma testosterone and CG development in castrated LS and HS quail were determined. Stress-line males were castrated and randomly allocated to 1 of 3 testosterone treatments: the empty testosterone (ET), low testosterone (LT), or high testosterone (HT) implant group. Cloacal gland volume was determined at 4 weekly intervals that represented ranges of 1 to 9 d, 8 to 17 d, 15 to 24 d, and 22 to 31 d after castration and testosterone implantation. Levels of plasma testosterone were also assessed at the end of the study. Development of the CG was affected by quail line (LS > HS), testosterone treatment (HT > LT > ET), and time of measurement (1 to 9 d < 8 to 17 d < 15 to 24 d = 22 to 31 d after castration and testosterone implantation). A significant interaction between testosterone treatment and time of measurement on CG volume was also detected (with CG volume generally increasing with time in LT- and HT-treated quail, but not in ET-treated quail). However, even though HT implant treatments induced higher CG development than did LT treatments beyond the first interval of CG volume measurement, and despite the finding of greater CG volumes in LS than HS quail during the last 2 measurement intervals within each of the LT and HT groups, no interaction was observed between testosterone implant dosages and quail stress line on CG volume. Thus, by the end of the study, regardless of testosterone dose, CG volume was consistently greater in LS quail than in their HS counterparts. In addition, although, as expected, the testosterone implant treatment significantly altered levels of plasma testosterone (HT > LT > ET), neither quail line nor its interaction with testosterone treatment affected plasma testosterone. The present findings suggest that the often-observed depressed CG development in the HS line may be independent of testosterone effects.

Plasma corticosterone responses to handling in Japanese quail selected for low or high plasma corticosterone responses to brief restraint.

1. Plasma corticosterone responses to handling in Japanese quail selected for low or high corticosterone responses to brief mechanical restraint (low and high stress lines) were measured in this study. 2. Initial corticosterone concentrations did not differ between the two lines. Corticosterone concentrations increased 15 min after handling began in quail handled for 5 or 15 min, with greater increases in quail from the high stress than the low stress line. Integrated corticosterone responses were higher in high stress than low stress quail, although the difference between the lines was not significant in quail handled for 5 min. 3. Patterns of plasma corticosterone after 15 min differed between quail handled for 5 or 15 min. Corticosterone increased further or did not change from 15 to 30 min then decreased from 30 to 60 min in quail handled for 5 min, whereas corticosterone decreased from 15 to 30 min then did not change in quail handled for 15 min. Corticosterone concentrations at 60 min remained elevated above initial concentrations, and were similar in quail handled for 5 or 15 min. Corticosterone concentrations at 15 min were more than twice as high in quail handled for 15 min compared with quail handled for 5 min. 4. Although the lines of quail have been selected on the magnitude of corticosterone responses, there was still marked variation in responses between birds within each line. Some quail in the high stress line showed little or no corticosterone response to handling, whilst large responses were seen in some birds of the low stress line. The lines of quail differ in characteristics other than corticosterone responses, including fear behaviour and reproductive development, so the lines provide valuable opportunities to investigate the potential significance of differences between individual birds in their corticosterone responses.

Short-term spatial memory responses in aged Japanese quail selected for divergent adrenocortical stress responsiveness.

Stress-induced glucocorticoids can dampen learning and spatial memory via neuronal damage to the hippocampus. Cognition losses can be transient (associated with acute stress episodes) or permanent as in aged individuals who show chronic glucocorticoid-induced accelerated brain aging and neurodegeneration (dementia). Thus, chronic versus acute stress effects on spatial memory responses of quail selected for reduced (low stress, LS) or exaggerated (high stress, HS) plasma corticosterone (B) response to brief restraint were assessed. Aged food-motivated male LS and HS quail were tested for 10 min in a feed-baited 8-arm radial arm maze (RAM) 1) at 255 d of age (quail who had experienced lifelong management stressors but who were otherwise never intentionally stressed; that is, chronically stressed birds), 2) on the next day post-acute stressor treatment (5 min of restraint), and 3) on the next day without treatment (acute stress recovery). The RAM tests used the win-shift procedure in which visited arms were not rebaited. Radial arm maze performance was measured by determination of the total number of arm choices made, the number of correct entries made into baited arms out of the first 8 choices, the time required to make a choice, and the number of pellets eaten. Line effects (P < 0.001 in all cases) were detected for total number of arm choices made (HS < LS), number of correct entries made into baited arms out of the first 8 choices (HS < LS), time required to make a choice (HS > LS), and number of pellets eaten (HS < LS). However, neither the effects of day of RAM testing nor its interaction with line further influenced these variables. Thus, although selection for divergent plasma B responsiveness to an acute stressor was found to be associated with severe impairment of spatial memory in aged male HS compared with LS quail, the observed spatial memory impairments (HS > LS) could not be further altered by acute stressor treatment. Line differences in cognition may reflect lifelong management-induced stress episodes that periodically produce higher plasma B responses in HS than LS quail, which underlie HS quail memory deficits, or other etiologies, or both.

Influence of maternal corticosterone treatment on incubation length of eggs laid by Japanese quail hens selected for divergent adrenocortical stress responsiveness.

1. Previous studies have shown that more yolk corticosterone is found in the eggs of random bred Japanese quail hens implanted with corticosterone during egg formation; both unstressed and stressed quail hens selected for exaggerated (high stress) rather than reduced (low stress) plasma corticosterone response to brief restraint deposit more corticosterone into their egg yolks. The length of egg incubation is also known to be shorter in eggs laid by high than low stress hens. 2. Here we investigated the interactive effects of quail stress line (low vs. high stress) with maternal corticosterone treatment (empty implant controls vs. corticosterone-implants) during egg formation on length of egg incubation. 3. Mean (+/-SEM) length of egg incubation for high stress control eggs (3973 +/- 04 h) was similarly shorter (by about 4.5 h) than that found for low stress control eggs (3928 +/- 02 h). In addition, on average, the incubation length of eggs laid by corticosterone-implanted hens (3929 +/- 05 h) was nearly 3 h shorter than that found for eggs laid by control hens (3958 +/- 02 h) regardless of stress line. 4. Line x hen-implant treatment effects on mean (+/-SEM) length of egg incubation partitioned in rank order as follows: low stress control (3978 +/- 05 h) > low stress corticosterone-implant (3959 +/- 07 h) > high stress control (3938 +/- 03 h) > high stress corticosterone-implant (3912 +/- 04 h). 5. Our original contention that selection for exaggerated adrenocortical responsiveness is associated with a reduction in the length of egg incubation was supported. Because maternal stress-induced elevations of yolk B are known to occur, the present findings of further shortenings of the hatching times of eggs of corticosterone-treated hens of both stress lines are also important to the poultry industry because they warn producers that unless stress in hens during egg formation is minimised, abbreviated egg incubation periods may result beyond the effects that a hen's genetic predisposition to adrenocortical stress responsiveness has on the length of egg incubation.

Effect of the density of conspecifics on runway social reinstatement behavior of male Japanese quail genetically selected for contrasting adrenocortical responsiveness to stress.

Runway tests are considered indicative of underlying sociality in birds and their ability to make social discriminations and establish interactions among conspecifics. Herein, social reinstatement behavior in male juvenile Japanese quail selected for a reduced (LS, low stress) or exaggerated (HS, high stress) adrenocortical response to brief mechanical restraint was evaluated. Individual males were given the choice to reinstate with either 2 (low density, LD) or 8 (high density, HD) unfamiliar randombred conspecifics placed in goal boxes at opposite ends of a two-choice runway (TCRW). Then, the same males were individually retested in a single goal box runway wherein they were exposed to a goal box containing either a LD or a HD of males. In the TCRW, a higher (P < 0.01) number of HS males started their ambulation toward the goal box containing the HD as opposed to LD of conspecifics. The HS males also spent more (P < 0.01) time in close proximity (within a 10-cm close zone; CZ) to the HD (218 s) rather than LD (57 s) of conspecifics. In contrast, the LS males did not differ in their initial direction of travel and they spent similar average amounts of times in the CZ of their stimulus LD (141 s) and HD (124 s) conspecifics. Similar to the TCRW results, in the single goal box runway, HS males spent more (P < 0.01) time in the CZ of HD rather than LD conspecifics, whereas LS quail spent similar amounts of time in the CZ of LD and HD males. Considering that runways are novel (and therefore frightening) environments, our findings suggest that HS quail may find better shelter (i.e., more comfort) in close proximity to a larger rather than smaller group of conspecifics, whereas LS birds find groups of varying conspecific density equally attractive. The results suggest that LS quail possess favorable social adaptive qualities because they appear to be better suited to cope with situations where the density of conspecifics is variable.

Maternal corticosterone reduces egg fertility and hatchability and increases the numbers of early dead embryos in eggs laid by quail hens selected for exaggerated adrenocortical stress responsiveness.

Quail hens selected for exaggerated (HS, high stress) rather than reduced (LS, low stress) plasma corticosterone (B) response to brief restraint deposit more B into their egg yolks than do LS hens. Female progeny of HS hens implanted with B also show reduced egg production when compared with female offspring of LS- and HS-control and LS-B-implanted hens. Herein, LS and HS hens were implanted (s.c.) with empty (controls, CON) or B-filled silastic tubes to assess the interactive influences of maternal B-treatment with quail stress line on egg fertility (FERT), total egg hatchability (TOTHATCH) and fertile egg hatchability, and the percentages of embryonic mortality (early dead, ED; late dead) and pipped eggs. Mean FERT was dramatically reduced in eggs of HS compared with LS hens and B-implanted compared with CON-treated hens (P < 0.0001, both cases). Line x implant treatment FERT outcomes partitioned (P < 0.05) as follows: LS-B = LS-CON > HS-CON > HS-B. In addition, TOTHATCH was also affected by line (LS > HS; P < 0.0001) and implant treatment (CON > B-implant; P < 0.0002) and line x implant treatment TOTHATCH means differed (P < 0.05) as follows: LS-CON = LS-B = HS-CON > HS-B. Fertile egg hatchability was reduced (P < 0.05) in HS-B-treated hen eggs when compared with LS-B and HS-CON hen eggs and more (P < 0.05) ED embryos were found in eggs laid by HS-B-implanted hens than in any other treatment group. Late dead and pipped egg percentages were unaffected by any treatment. The findings are important to avian geneticists because they further emphasize the benefits that selection for reduced adrenocortical responsiveness has on hen reproductive performance. The maternal B findings also warn poultry and hatchery managers that unless hen stress during egg formation is avoided, negative consequences in FERT, TOTHATCH, and ED can result, particularly in hens genetically predisposed toward exaggerated adrenal stress responsiveness.

Fractal analysis of the ambulation pattern of Japanese quail.

1. The study examined the practicality and usefulness of fractal analyses in evaluating the temporal organisation of avian ambulatory behaviour by using female Japanese quail in their home boxes as the model system. To induce two locomotion activity levels, we tested half of the birds without disturbance (Unstimulated) and the other half when food was scattered on the floor of the home box after 3 h of feeder withdrawal (Stimulated). 2. Ambulatory activity was recorded during 40 min at a resolution of 1 s and evaluated by: (1) detrended fluctuation analyses (DFA), (2) the frequency distribution of the duration of the walking or non-walking events (FDD-W or FDD-NW, respectively), and (3) the transition probabilities between walking/non-walking states. Conventional measures of total time spent walking and average duration of the walking/non-walking events were also employed. 3. DFA showed a decreased value of the self-similarity parameter (alpha; indicative of a more complex ambulatory pattern) in Stimulated birds compared to their Unstimulated counterparts. The FDD-NW showed a more negative scaling factor in Stimulated than in Unstimulated birds. Stimulated birds also had more transitions between non-walking and walking states, consistent with stimulated exploratory activity. No differences were found between groups in the FDD-W, in percentage of total time spent walking, or in average duration of the walking events. 4. The temporal walking pattern of female Japanese quail has a fractal structure and its organisation and complexity is altered when birds are stimulated to explore. The fractal analyses detected differences between the Unstimulated and Stimulated groups that went undetected by the traditional measurements of the percentage of total time spent walking and the duration of the walking events suggesting its usefulness as a tool for behavioural studies.

Length of incubation and hatchling body weight in quail selected for reduced or exaggerated adrenocortical responsiveness to brief restraint.

1. While changes in corticosterone (B) during late embryogenesis are thought to play a role in the hatching process, only a scant and controversial literature exists concerning the effects of in ovo B on the length of incubation and chick body weight at hatching. Because female Japanese quail selected for exaggerated (high stress, HS) rather than reduced (low stress, LS) plasma B response to brief mechanical restraint deposit more B into their eggs, these stress lines provide an excellent model to study the relationship between embryonic B and incubation length and hatchling body weight. 2. Differences in the duration of incubation and chick body weight at emergence were assessed using eggs laid by LS and HS quail. 3. On average, eggs from HS hens hatched 3.7 h sooner than did eggs from LS hens while mean body weight at emergence was similar in hatchlings of both lines. Thus, selection for exaggerated adrenocortical responsiveness is associated with a reduction in the length of egg incubation without altering hatchling body weight. 4. This finding is important to the poultry industry because it warns layer, breeder farm and hatchery managers that unless stress in hens during egg formation is avoided, an abbreviated incubation period may result.

Open-field temporal pattern of ambulation in Japanese quail genetically selected for contrasting adrenocortical responsiveness to brief manual restraint.

Japanese quail selected for a low-stress (LS), rather than a high-stress (HS), plasma corticosterone response to brief restraint have been shown to possess lower fearfulness and a nonspecific reduction in stress responsiveness. Detrended fluctuation analysis provides information on the organization and complexity of temporal patterns of behavior. The present study evaluated the temporal pattern of ambulation of LS and HS quail in an open field that represented a novel environment. Time series of 4,200 data points were collected for each bird by registering the distance ambulated every 0.5 s during a 35-min test period. Consistent with their known reduced fearfulness, the LS quail initiated ambulation significantly sooner (P < 0.02) and tended to ambulate more (P < 0.09) than did their HS counterparts. Detrended fluctuation analyses showed a monofractal series (i.e., a series with similar complexity at different temporal scales) in 72% of the birds. These birds initiated their ambulatory activity in less than 600 s. Among these birds, a lower (P < 0.03) autosimilarity coefficient (alpha) was found in the LS quail than in their HS counterparts (alpha = 0.76 +/- 0.03 and 0.87 +/- 0.03, respectively), suggesting a more complex (less regular) ambulatory pattern in the LS quail. However, when the patterns of ambulation were reexamined by considering only the active period of the time series (i.e., after the birds had initiated their ambulatory activity), monofractal patterns were observed in 97% of the birds, and no differences were found between the lines. Collectively, the results suggest that during the active period of open-field testing, during which fear responses are likely less strong and other motivations are the driving forces of ambulation, the LS and HS lines have similar ambulatory organization.

Influences of maternal corticosterone and selection for contrasting adrenocortical responsiveness in Japanese quail on developmental instability of female progeny.

Differences in developmental instability were assessed in female offspring of Japanese quail hens selected for reduced (low stress, LS) or exaggerated (high stress, HS) plasma corticosterone (B) response to stress and treated with a placebo or B during egg formation. Hens of each line were implanted (s.c.) with either a silastic tube containing no B (controls) or one filled with B. Female chicks hatched from each of the 4 line x implant treatment combinations were retained for examination of 3 bilateral traits at 130 d of age: length of the tibiotarsus, middle toe length, and distance between the auditory canal and the nares (face length, FL). Greater bilateral trait size variances were associated with measurement of tibiotarsus length (P < 0.04) and middle toe length (P < 0.06) in the HS line, supporting our previous findings in the opposite sex that developmental instability (i.e., fluctuating asymmetry, FA) of certain morphological traits is more pronounced in HS than LS adult quail. The HS quail are also known to exhibit greater adrenocortical responsiveness to a wide range of stressors, and they are more easily frightened than LS birds. Therefore, the line differences in FA (HS > LS) found previously in males and herein in females may simply reflect the differential responsiveness of the birds to chronic social and physical environmental stressors. In addition, the present study detected more (albeit marginally so, P < 0.06) bilateral variability (i.e., heightened FA) in FL of quail hatched from mothers treated with B, a finding entirely due to the very high FL variance observed in the female offspring of B-treated HS hens. Because others have found in ovo B treatment to be associated with heightened FA in chick tarsus bone length and because we have also demonstrated that greater yolk B deposition occurs in eggs from both unstressed and stressed HS quail hens than their LS counterparts, the present maternal B treatment may be acting independently, or in combination with HS genomic effects, to adversely affect developmental stability.

Fear responses of offspring from divergent quail stress response line hens treated with corticosterone during egg formation.

Increased fearfulness has been associated with adrenocortical activation. Maternal corticosterone (B) treatment increases egg B, and elevated B in ovo enhances chick avoidance of humans. Quail selected for exaggerated (high stress, HS) rather than reduced (low stress, LS) plasma B response to stress are more fearful, and more B is found in HS hen eggs. Thus, we used tonic immobility (TI) and hole-in-the-wall box (HWB) emergence tests to assess fear in chicks hatched from eggs of LS and HS hens implanted with B or no B (CON). The number of inductions required to attain TI, latency to first alert head movement, and duration of TI were determined in one study and the latency until first vocalization (LATVOC), numbers of vocalizations (VOCS), proportions of chicks vocalizing, and the latencies to head (HE) and full-body (FE) emergence from a HWB were assessed in another. The LS chicks required less inductions (P < 0.0005) and had shorter latency to first alert head movement (P < 0.02) than HS chicks, although the duration of TI was unaffected by any of the treatments. During the acclimation period of the HWB tests, more (proportions of chicks vocalizing; P < 0.0001) HS chicks alarm-called sooner (LATVOC; P < 0.0001) and more often (VOCS; P < 0.0001) than did LS chicks, and, although maternal implant treatment did not affect LATVOC, progeny of B-implanted hens showed a tendency toward less (P < 0.07) VOCS than the CON. Chicks hatched from eggs of B-implant mothers also took longer to achieve HE (P < 0.06) and FE (P < 0.05) from the HWB than did their CON counterparts. Stress line, implantation treatment, and their interaction did not alter HE or FE responses. The data suggest that quail stress line genome may or may not be affecting certain fear and alarm responses in chicks via the same mechanism(s) that underlies how elevating maternal B increases egg levels of B that in turn alters the fear behavior of progeny.

Ontogeny of the cloacal gland in male Japanese quail classified in a T-maze.

Broiler chicks that traverse a T-maze quickly to reinstate contact with their companions (HP, high performers) are known to grow faster, are more social, and have a reduced plasma corticosterone response to acute stress than slower chicks (LP, low performers). High-performing quail from a line selected for reduced rather than exaggerated (high-stress) adrenocortical stress responsiveness also show enhanced female reproductive performance when compared with LP-high-stress quail. Herein, time courses of male sexual development were evaluated in genetically unremarkable quail that were categorized at 2 d of age as HP or LP in the T-maze. Body weight, cloacal gland volume (CVOL), proportion of individuals that produced cloacal gland foam, intensity of foam production, and CVOL relative to BW (RCVOL) were determined weekly from 4 to 10 wk of age, and again at 22 wk, along with absolute and BW-adjusted testes weight. Although CVOL and RCVOL were initially similar in both T-maze groups, both variables were greater (P < 0.05) in HP than in LP quail between 6 and 10 wk of age. High-performing birds also showed a trend (P < 0.1) of greater cloacal gland foam than their LP counterparts from 5 to 7 wk. From 8 wk on, all birds were in foam production. Intensity of foam production results generally mimicked those found for CVOL and RCVOL. Body weights were higher (P < 0.05) in HP than LP quail from 5 to 7 wk. No T-maze group differences were found in any of the variables at 22 wk of age. The results suggest that rapid negotiation of the T-maze is associated with accelerated growth and puberty in male quail, although the enhanced reproductive development of HP males does not remain extant in aged adults.

Cloacal gland growth differences in high and low plasma corticosterone stress response line male quail reared under short daylengths.

Mail quail selected (generation 32) for reduced [low stress (LS)] or exaggerated [high stress (HS)] plasma corticosterone stress response to brief mechanical restraint (5 min of immobilization) were studied for differences in the growth of their cloacal glands when reared essentially lifelong on short day lengths of 8L:16D. Post-brooding, at 4 wk of age, 96 quail (48 LS+48 HS) were housed in cages (1 LS and 1 HS male/cage), and the short day light treatment was instigated. Using a digital caliper, cloacal gland (CG) size measurements (length and width, mm) were made biweekly beginning at 42 d of age and ending at 196 d of age (28 wk old). The CG volume (CVOL) was calculated from these measurements using a literature-proposed formula. The CVOL in both lines increased similarly with increasing age from 6 to 12 wk of age. However, beginning at 14 wk of age and thereafter, LS males showed greater (P<0.05) CVOL than HS ones. No further increases in CVOL were observed in either line at 24 wk of age beyond that seen at 22 wk. This study demonstrated that although both lines show CG development under short days, eventually CG growth becomes comparatively stymied in the HS males. Furthermore, the maintenance of similarly lower CVOL in HS than LS males, CVOL that showed no further increase from 22 to 24 wk of age, suggests that holding these quail lifelong on short days results in, comparatively, yet another permanent negative reproductive consequence in quail selected for exaggerated plasma corticosterone stress responsiveness.

Maternal corticosterone further reduces the reproductive function of male offspring hatched from eggs laid by quail hens selected for exaggerated adrenocortical stress responsiveness.

Activation of the hypothalamic-pituitary-adrenal (HPA) axis can depress the hypothalamic-pituitary-testicular axis. Male quail cloacal gland (CG) size and foam production shows androgen dependency, and males selected for exaggerated [high stress (HS)] rather than reduced [low stress (LS)] plasma corticosterone (B) stress response exhibit reduced CG and testes development. High stress hens also deposit more B into egg yolks than LS ones, and quail hens given B produce chicks that have a reduced growth rate and adults with heightened HPA responsiveness. Herein, we gave LS and HS hens no B [empty implants, control (CON)] or B-filled implants and assessed the reproductive performances of these hens and their male offspring. Mortality was similarly elevated in LS and HS B-treated hens, but only HS B-implanted hens showed reduced egg production. In male offspring, CG volume (CVOL), intensity of CG foam production (CFP), and the proportion of individuals that produced CG foam were measured from 4 to 11 wk of age. At 6 wk, BW, and at 15 wk, BW, testes weight (TWT), and TWT relative to BW were also determined. Hen treatments did not affect male chick CVOL at 4 wk, but CVOL differed thereafter as follows: LS CON > LS B = HS CON = HS B at 5 and 6 wk and LS CON > LS B > HS CON = HS B from 7 to 11 wk. By 8 wk, and thereafter, CFP differed as follows: LS CON > LS B > HS CON > HS B. Group differences in the proportion of individuals that produced CG foam generally supported CFP findings from 4 to 8 wk of age. Body weight did not differ by treatment at 6 wk of age. By 15 wk, TWT were similarly depressed in both HS groups. However, similarly higher 15-wk BW in the LS-CON and HS-B groups contributed to TWT relative to BW differences as follows: LS-B > LS-CON > HS-B; LS-CON = HS-CON; LS-B > HS-CON; and, HS-CON = HS-B. Both selection for exaggerated HPA responsiveness and maternal B treatment negatively affected the reproductive function of HS male offspring.

Cloacal gland and gonadal photoresponsiveness in male Japanese quail selected for divergent plasma corticosterone response to brief restraint.

Corticosterone is linked to testicular depression, and in Coturnix, a relatively photorefractory species, day length and androgen dependency in cloacal gland development and foam production are evident. Furthermore, male quail selected for reduced (low stress, LS) rather than exaggerated (high stress, HS) plasma corticosterone stress response show more photo-induced reproductive development, greater resistance to reproductive declines induced by exposure to very short days, and a quicker recovery to a higher reproductive level upon return to long days. To determine whether a milder reduction in day length would also influence stress line reproductive photoresponsiveness, and perhaps photorefractoriness, males grown on 16 h of light were given 13 h of light for 4 wk followed by a return to long days for 12 wk. Cloacal gland measurements were made following the initial stimulatory photoperiod and weekly thereafter during the 2 light treatments. Plasma testosterone was determined initially, after 4 wk of light reduction, and at the end of the study, along with BW, testes weight (TWT), and TWT relative to BW. Cloacal gland volume (CVOL) was greater in LS than HS males grown on long days. Whereas exposure to 13 h of light reduced CVOL in both lines, line differences (LS > HS) persisted during the first 3 wk of light reduction. Moreover, by 2 wk of rephotostimulation, and weekly thereafter, line differences (LS > HS) in CVOL reemerged. Cloacal gland foam production and the proportion of individuals that produced cloacal gland foam responses reflected changes in CVOL. The BW did not differ by line, yet LS males had a higher TWT and TWT relative to BW than HS ones. All individuals, however, exhibited relative photorefractory responses. Whereas none of the LS males completely resisted reproductive regression induced by a 3 h light reduction, LS males showed other reproductive benefits upon mild photocastration and subsequent rephotostimulation (e.g., greater resistance to CVOL and cloacal gland foam production losses during a mild light crash and quicker restoration to full reproductive potential upon relighting).

Active immunization of broiler breeder cockerels against chicken inhibin accelerates puberty and prevents age-induced testicular involution.

Injection of quail and breeder hens with a recombinant protein antigen (MBP-cINA521)--a fusion of the bacterical maltose-binding protein (MBP) and a fragment of the alpha-subunit of chicken inhibin (cINA521)--accelerates puberty and enhances lay. Herein, the effects of this immunogen on reproductive responses in broiler breeder males were assessed. Cockerels were subcutaneously injected with 0 (vehicular controls), 1, 3, or 5 mg of MBP-cINA521 at 13 wk of age and with one-half of these dosages (boosters) at 18 wk. Bird subsamples were weighed, blood sampled, and killed at 24, 28, and 39 wk of age to assess age and vaccination effects on BW, testes weight (TWT), TWT relative to BW (RTWT), TWT > or = 20 g (TWT20; theoretical threshold TWT for maximum fertility), and plasma testosterone. Breeder males are sexually developing, reach peak sexual activity, and show age-related reproductive decline at these ages. Because vaccine gonadal effects at 24 wk appeared to be dramatic, the size of the left testis was also scored to see if size differences could be detected by mere visual inspection. Male fighting increasingly reduced sample sizes beyond 24 wk. Because mortality was unrelated to the treatments and to insure meaningful statistical comparisons, MBP-cINA521 data were pooled. Body weight (P < 0.04), testis score (P < 0.02), TWT (P < 0.03), RTWT (P = 0.06), and plasma testosterone (P = 0.08) were elevated in immunogen-treated males at 24 wk of age, and more (P < 0.05) MBP-cINA521-treated birds than controls achieved a TWT20 at this time. These variables did not differ by treatment at 28 wk. However, by 39 wk, treatment effects reemerged as follows: TWT (P < 0.04), RTWT (P = 0.06), and TWT20 (P < 0.01) were increased in vaccinated males who also showed nearly 3-fold higher levels of plasma testosterone. We conclude that immunoneutralization of inhibin accelerates puberty and retards age-related sexual senescence that typically occurs in broiler breeder males.

Stressor-induced changes in open-field behavior of Japanese quail selected for contrasting adrenocortical responsiveness to immobilization.

The effect of no treatment (undisturbed controls; CON) or 5 min exposure to immobilization stress (STR) before assessment of open-field behavior in Japanese quail lines selected for high (high stress; HS) or low (low stress; LS) plasma corticosterone (B) response to a similar immobilization stress was examined. During a 5-min test period in an open field, the following behavioral measurements were made: the latency to ambulate (LAMB), the number of open-field floor sectors entered (SECTORS), and freezing behavior (total time spent in complete silence and inactivity, apart from slight movements associated with respiration; FREEZE). A further measure of ambulation across time (ambulation rate; ARATE) was calculated using the formula: ARATE = (SECTORS/[(300 s test ceiling)--LAMB)]. The LS-quail showed higher (P < 0.04) ARATE and lower (P < 0.05) FREEZE behavior than did their HS counterparts. When compared with the CON, quail exposed to STR showed decreased ARATE (P < 0.02) and numbers of open-field SECTORS entered (P < 0.04). More importantly, the LS-CON exhibited a higher (P < 0.05) mean ARATE, less FREEZE behavior, and increased numbers of SECTORS entered than did the other 3 treatment groups (LS-STR, HS-CON, and HS-STR), all of which showed similar responses for these variables. The LS-CON vs. HS-CON findings support our contention that selection for reduced adrenocortical responsiveness has been accompanied by a concomitant decrease in underlying fearfulness (exemplified herein by heightened activity in the open field in LS-CON). We further show here that this relationship was eliminated by application of an acute stressor before open-field testing of LS quail, wherein a reduction in activity similar to that found in both HS quail treatment groups was evident in comparison to the LS-CON.

Adrenocortical responses of Japanese quail to a routine weighing procedure and to tonic immobility induction.

Tonic immobility (TI) is induced by brief manual restraint, and it is a commonly used test of fearfulness, particularly in poultry. However, in view of increasing ethical concerns about experimentation on living animals, there is a need to ensure that the tests used do not elicit unacceptable distress. In the United Kingdom, there is some debate as to whether TI should be classified as a regulated experimental procedure that requires a Home Office license to experiment on living animals under the Animals (Scientific Procedures) Act of 1986. The present study compared plasma corticosterone (PC) concentrations in undisturbed Japanese quail (controls) with those exposed to the induction of TI or those exposed to a routine weighing procedure. Circulating PC concentrations were higher following TI induction or weighing than in the controls, but the 2 treatments elicited similar adrenocortical responses. Further, the PC levels found here were lower than those reported elsewhere when quail were exposed to crating, transport, or mechanical immobilization stressors. We therefore concluded that the induction of TI might be best regarded as a mild stressor. Furthermore, studies using this technique can identify ways of alleviating fear and thereby improving well-being. Collectively, we believe that the use of TI is justified, and it does not require classification as a regulated procedure in the United Kingdom or elsewhere.