Comb size, shape complexity and laterality of laying hens reared in environments varying in resource choice.

The comb is an ornament involved in signalling condition in domestic fowl. We hypothesised that comb size, comb shape complexity (i.e., rugosity, the comb perimeter jaggedness), and comb laterality of laying hens would be influenced by the degree of environmental enrichment experienced during juvenile development in the form of resource choice. We conducted a 2 × 2 factorial crossover experiment with pullets reared in pens containing four perches of equal length and four litter areas of equal size. Pullets were exposed to a single choice vs multiple choices of perch and litter types (i.e., all the same vs all different) during Weeks 1-4 (Period 1) and/or Weeks 5-15 (Period 2) of rearing (n = 4 pens/treatment combination) prior to transfer to standard adult laying pens for Weeks 16-27 (Period 3). In Week 27, combs were photographed, and comb laterality (hanging on left or right side) was noted. Using a custom-made image analysis programme, we captured comb area (mm2), perimeter length (mm), and rugosity ((perimeter length - horizontal length) / horizontal length) from comb photographs of 6-7 randomly selected hens/pen. We predicted that hens reared in the multi-choice environment during Periods 1 and 2 would have larger, more complex, and left-side-biased combs than those in the other treatment groups, reflecting lower allostatic load. The predicted comb side bias was based on a possible bias in head posture/movements associated with greater right eye/ear use and left-brain hemispheric dominance. Contrary to our predictions, we detected an overall right-side bias in comb laterality, and no associations between resource choice treatment in Period 1 or Period 2 and comb area, perimeter length, rugosity, or laterality of the adult hens. Thus, variation in allostatic load resulting from the rearing treatments was insufficient to modify the trajectory of comb morphological development, possibly due to a ceiling effect when comparing environmental treatments on the positive end of the welfare spectrum. We found that left-lopping combs had shorter perimeters than right-lopping combs. However, among hens with left-lopping combs, those with larger combs were heavier and had less feather damage, while among hens with right-lopping combs, those with longer-perimeter combs were heavier and tended to have less comb damage. In conclusion, comb characteristics were related to physical condition at the individual level but did not serve as sensitive integrated indicators of hen welfare in response to basic vs enhanced resource choice during rearing.

The ability of White Leghorn hens with trimmed comb and wattles to thermoregulate.

It is estimated that each year over 19 million pullets in the United States have their combs partially trimmed at a young age to improve egg production and feed efficiency. A possible disadvantage of trimming is that the comb and wattles may be essential for thermoregulation during hot weather allowing for conductive cooling of the blood through vasodilation of superficial vessels in these integumentary tissues. The objective of this study was to investigate the effects of partial comb and wattle removal, performed at 21 d of age, on the ability of White Leghorns to thermoregulate before, during, and after an imposed heating episode that averaged 34.6°C for 50.5 h. An increase in mortality at 20 h and body temperature at 48 h post initiation of the heating episode demonstrated that hens with trimmed comb and wattles were not able to cope with heat stress as effectively as controls. The increase in wattle temperature in controls as compared to trimmed hens during the heating episode and following heat stress provides supportive evidence that blood pools to the peripheral surface for conductive heat loss. During high temperatures typical of summer, trimmed hens attempted to compensate for their lack of ability to transfer heat from their comb and wattles to the environment through increased proportion of panting and wing spreading. Under less extreme conditions with lowered ambient temperatures, the trimming of the comb and wattles did not impair the ability of hens to thermoregulate, as body temperatures and behavior were similar to controls with no mortality. Egg weight was the only production parameter adversely affected by the trimming of the comb and wattles as compared to controls. The implication is that hens need their combs and wattles to thermoregulate effectively during periods of high environmental temperature. Pullets should not be subjected to a comb and wattle trim if they are housed in laying facilities that are not appropriately cooled during the summer.

Effect of partial comb and wattle trim on pullet behavior and thermoregulation,1.

The wattles and comb of chickens are important for thermoregulation allowing for heat exchange during high temperatures. These integumentary tissues are sometimes trimmed to prevent tears if caught on cage equipment and to also improve feed efficiency; however, the procedure itself could be painful to chicks. Our objective was to determine the effect of trimming the comb and wattles on behavior, BW, feed usage, and the surface temperature of pullets. The wattles and comb of Leghorns were partially trimmed at 21 d age (n = 6 cages, 13 chicks each) with another 6 cages serving as controls. Behaviors were recorded 3 times daily for 1 h starting at 0800, 1200, and 1500 using instantaneous scan sampling observations conducted every 5 min prior to, on the d of, and after the trim. Group BW at 21, 28, and 36 d age and the amount of feed used for 7 d beginning at 21 and 29 d age were measured per cage. At 1300 h on d before and after the trim, thermal images of the pullet's beak, comb, eye, wattle, and shank were randomly taken on 3 pullets/cage. Smaller proportions of trimmed chicks were eating and greater proportions were sitting on the d of the trim as compared to the controls with the opposite trend occurring on 4 d post-trim (treatment by age interaction, P = 0.03 and 0.0001, respectively). Standing behavior differed only on the d of the trim where smaller proportions of trimmed pullets stood as compared to intact controls (treatment by age interaction, P = 0.0002). Trimming the comb and wattles did not affect preening, running, BW, feed utilization, and the surface temperature of the pullet. The indices of behavior suggest that on the d of the trim, pullets may have experienced temporary distress, but they returned to normal behavior by 5 h post-trim with no long-term effect on BW, feed usage, or surface body temperature.

Intra-sexual competition alters the relationship between testosterone and ornament expression in a wild territorial bird.

In a reliable signalling system, individual quality is expected to mediate the costs associated with ornamental displays, with relatively lower costs being paid by individuals of higher quality. These relative costs should depend not only on individual quality, but also on levels of intra-sexual competition. We explored the current and delayed effects that testosterone implants have on bird ornamentation in populations with contrasted population densities, as a proxy for intra-sexual competition. In a replicated experiment, we manipulated testosterone in 196 yearling male red grouse Lagopus lagopus scoticus in autumn in populations of high and low levels of intra-sexual competition. Males were assigned to one of three exogenous testosterone (T) treatments: empty implants (T0), small T implants (T1) or larger T implants (T2). We monitored subsequent changes in testosterone levels, ornament size and carotenoid-based colouration, carotenoid levels and body condition from autumn to spring. Testosterone implants increased testosterone levels, comb redness and comb size, and decreased body condition but these effects depended on levels of intra-sexual competition. Specifically, T2-implanted birds increased testosterone levels and comb size more, and reduced body condition more, in populations where intra-sexual competition was low. In the following spring, testosterone levels of T2-treated birds kept increasing in populations where intra-sexual competition was high but not in populations where intra-sexual competition was low. Our results highlight that levels of intra-sexual competition alter the relationship between testosterone levels and ornament expression, influencing their condition-dependence; they also indicate that the outcome of standard hormone manipulation conducted in free-living animals vary depending on the population context.

Peripheral temperature drop in response to anticipation and consumption of a signaled palatable reward in laying hens (Gallus domesticus).

The present study describes effects of anticipation and consumption of a palatable reward on comb surface temperature. The purpose was to investigate temperature responses as a potential physiological indicator of positive emotional states in laying hens. A rise in body temperature in response to stimuli predictive of or during exposure to unpleasant events has been interpreted as evidence of emotions in mammals and avians. However, this phenomenon has so far only been studied during anticipation of or exposure to negative events; i.e., emotions of a negative valence. Infrared thermography was used to record potential alterations in comb surface temperature to a conditioned cue signaling a reward (mealworms) and during reward delivery. On average, comb temperature dropped 1.5 °C (95% CI: +/-1.2 °C) after exposure to CS and consumption of reward (p~0.0014) when initial comb temperature was above 30 °C. Such temperature drop indicates a peripheral vasoconstriction and has clear resemblances to emotional fever as seen during negative emotional states. Thus, we propose that a drop in peripheral temperature reflects emotional arousal more than emotional valence. Substantial temperature responses due to diet-induced thermogenesis were found, further emphasizing a cautious interpretation of altered comb temperature in studies of animal welfare.

Condition- and parasite-dependent expression of a male-like trait in a female bird.

In many species, females display brightly coloured and elaborate traits similar to those that males use in intra- and inter-sexual selection processes. These female characters are sometimes related to fitness, and might function as secondary sexual characteristics that have evolved through sexual selection. Here, we used descriptive data from 674 females in 10 populations and an experimental removal of Trichostrongylus tenuis parasites in four populations, to examine the effects of season, age, condition, and parasites on the size of supra-orbital combs displayed by female red grouse Lagopus lagopus scoticus. We found that comb size (i) was greater during the breeding than the non-breeding season, (ii) was greater in adult than in young females, (iii) was positively correlated with body condition, and (iv) negatively correlated with parasite abundance. Experimentally, we showed that comb size increased proportionally to the number of worms removed after parasite dosing. Our findings provide a better understanding of proximate mechanisms behind the expression of a male-like trait in females, and we discuss its possible function as a female ornament.

Testing the interactive effects of testosterone and parasites on carotenoid-based ornamentation in a wild bird.

Testosterone underlies the expression of most secondary sexual traits, playing a key role in sexual selection. However, high levels might be associated with physiological costs, such as immunosuppression. Immunostimulant carotenoids underpin the expression of many red-yellow ornaments, but are regulated by testosterone and constrained by parasites. We manipulated testosterone and nematode burdens in red grouse (Lagopus lagopus scoticus) in two populations to tease apart their effects on carotenoid levels, ornament size and colouration in three time-step periods. We found no evidence for interactive effects of testosterone and parasites on ornament size and colouration. We showed that ornament colouration was testosterone-driven. However, parasites decreased comb size with a time delay and testosterone increased carotenoid levels in one of the populations. This suggests that environmental context plays a key role in determining how individuals resolve the trade-off between allocating carotenoids for ornamental coloration or for self-maintenance needs. Our study advocates that adequately testing the mechanisms behind the production or maintenance of secondary sexual characters has to take into account the dynamics of sexual trait expression and their environmental context.

Sequence analysis of a pea comb locus on chicken chromosome 1.

To facilitate gene identification, this study aimed to narrow the scope of the genome region affecting chicken comb type by using two bird populations. First, an F2 resource population was generated by crossing Japanese game fowl (Shamo; pea comb, P/p and P/P) with White Plymouth Rock (single comb, p/p). Comb types of the 240 F2 offspring produced by an F1 intercross between eight males and 57 females were segregated at a ratio of 3:1 (pea:single). The pea comb locus was mapped to a chromosomal region on Gallus gallus chromosome 1 that was flanked by microsatellite markers MCW0112, MCW0019 and ABR521. The second population (five-generation, n=1300 animals) was derived from a cross between Shamo and Rhode Island Red (single comb, p/p) that had been genotyped for additional polymorphic single nucleotide polymorphisms and microsatellite markers within this region through development of chicken draft sequences. To close some gaps in these draft sequences, we constructed a bacterial artificial chromosome contig and sequenced it using the shotgun sequencing technique. Chickens selected from pedigrees in these populations were grouped by inheritance of a P or p haplotype at the locus constructed by the additional markers. Finally, this locus was fine-mapped to roughly 60 kb based on the association of haplotypes and comb types. Chicken genome sequences suggest that the most likely polymorphism responsible for the pea comb locus is a duplicated sequence and that the sex determining region Y-box 5 gene, one predicted gene and one expressed sequence tag in a critical region may be associated with the duplicated sequence.

Color signals in the black grouse (Tetrao tetrix): signal properties and their condition dependency.

Recent progress in techniques of quantifying between-individual differences of color-based ornaments has revealed undiscovered possibilities for research in sexual selection. We present how the color spectra data can be comprehensively used for studying the importance of sexual ornaments in the black grouse and how these ornaments are related to a male condition. For this, we used both correlative field and experimental data. Field data indicated that older males had more chromatic coloration than yearlings. Blue chroma of males was correlated with male mating success. We experimentally manipulated yearling birds with testosterone implants and found that testosterone-implanted males had impaired expression of several sexual ornaments: 10 months after the implantation, both structural-based blue and carotenoid-based red eye comb coloration were diminished, as well as lyre (tail) length. However, the manipulation did not affect vital traits under natural selection (wing length or body mass). Our data indicate that structural color is an important trait in sexual selection in this lekking species. Importantly, the data also indicate that male sexual ornaments are more susceptible to environmental conditions than the other male traits, thus showing their heightened condition dependency compared with the other traits mediating the honesty of signaling.

Testosterone and autumn territorial behavior in male red grouse Lagopus lagopus scoticus.

In many bird species, males exhibit territorial aggression outside the breeding season, when testosterone concentrations are low and may not regulate territorial behaviors. The hormonal regulation of aggression at this time of year has only been studied in passerine birds. Here, we investigated the role of testosterone in the regulation of aggression in a non-passerine bird, the red grouse Lagopus lagopus scoticus. Male red grouse are aggressive in early spring when breeding starts, in autumn when they establish territories, and sporadically through much of the winter. We first describe seasonal variations in plasma testosterone concentrations and in the size of males' sexual ornaments, their red combs, which relates to aggressiveness. Testosterone concentrations and comb size were correlated. Both increased in autumn to a peak in October, and then increased again in spring, to a greater peak in early April. Secondly, we experimentally investigated the effects of testosterone, and of an anti-androgen (flutamide) used in combination with an aromatase inhibitor (ATD), on autumn territorial behavior. Males were treated with either empty implants, as controls (C-males), testosterone implants (T-males), or with flutamide and ATD implants (FA-males). One month after implanting, both T- and FA-males had higher concentrations of testosterone than C-males. Comb size, aggressive call rate, and response to playbacks of territorial call all significantly increased in T-males. However, the increase in testosterone in FA-males did not increase comb size or aggressive behavior. In the following spring, after the content of implants was used, FA-males had significantly lower testosterone than C-males, and had a reduced seasonal increase in comb size. The results suggest that testosterone plays a significant role in regulating red grouse aggressive behavior in autumn. However, the observation that flutamide and ATD treatment did not reduce territorial behavior, suggests that estradiol may also be involved in the regulation of non-breeding aggression.

[The effect of the husbandry system on comb size and comb color in hens]. / Einfluss der Haltung auf Kammgrösse und Kammfarbe bei Hühnern.

The comb size and comb colour of 84 hens (white and brown Hisex), living in battery cages or at free range were investigated. The main question was: does the comb size and colour correlate with the keeping systems i.e. with the quality and intensity of light and the ambient temperature? The results confirm these hypotheses and also show the genetic influence. The crest size of Hisex white hens is larger and paler compared with those of Hisex brown hens. This tendency is more distinct under the conditions of artificial light, less light and higher temperature of the environment in battery cage systems. Comb size and colour are used as a criterium for health and egg production of the hens. The combs are very important for the hen's health for the regulation of the body temperature and for absorbing light and also for the social structure of the flock; the comb form is an essential characteristic to identify the individual animal.

Neurogenic inflammation in the chicken (Gallus gallus var domesticus).

1. Neurogenic inflammation has been studied in the anaesthetized adult hen using a variety of different stimuli. 2. Plasma extravasation was produced following antidromic stimulation of the external mandibular ramus of the trigeminal nerve which innervates the skin at the angle of the jaw and the anterior part of the wattle. 3. Stimulation of the wattle by external application of mustard oil, thermal and mechanical stimuli, as well as intradermal injection of substance P and bradykinin, all produced plasma extravasation. 4. These results demonstrate that, in contrast to previous findings in the pigeon, at least in the trigeminal of the chicken peripheral C-fibre nociceptors have similar physiological characteristics in relation to the neurogenic inflammatory mechanism to those seen in mammals.

Tissue resistivities and current pathways and their importance in pre-slaughter stunning of chickens.

The failure of commercial pre-slaughter electrical stunning to stun effectively all birds may be, in part, a result of the current following slightly different paths through the tissues of different birds. Resistivity can be used to predict current pathways, and methods are presented for determining the resistivity of a tissue in freshly-killed or anaesthetized adult fowls. A variety of tissues were tested including comb, skeletal muscle, epithelium, brain, skull bone, heart and liver. The resistivity of a particular tissue varied little between birds, with the exception of skull bone in which resistivity differed markedly from bird to bird. This variation in skull bone resistivity may explain why some birds are inadequately stunned, since in these individuals the current density in the brain is likely to be insufficient to induce an adequate epileptic seizure.

Chick-comb fibroblasts. A new strain of cultured cells sensitive to androgens.

An androgen-responsive cell strain of chick-comb fibroblasts has been established in culture. Cells were obtained from combs of 1--2 day old white Leghorn cockerels and cultured in Dulbecco's modified Eagle medium supplemented with 10% fetal calf serum and 6 g/l glucose. The cells required serum but not androgens for growth. In the presence of androgens, growth rate and incorporation of labeled precursors into protein and RNA were markedly increased in exponentially growing cells, but not in confluent cultures. Androgens exerted these stimulatory effects in a dose-dependent fashion. It is concluded that chick-comb fibroblasts, which respond to androgens in vivo, retain this responsiveness when cultured. They furnish a well-defined system of potential value for studies of the molecular mechanisms of androgen action.