James R. Millam

The importance of mate behavioural compatibility in parenting and reproductive success by cockatiels, Nymphicus hollandicus

In species with long-term pair relationships, behavioural compatibility between mates could be a significant source of selection among pairs. We explored the relation between behavioural compatibility and reproductive success in cockatiels, a socially monogamous species with biparental care and variable compatibility. The mate behavioural compatibility measure, derived from our previous work with these birds, was based on five pair-level variables: proximity, behavioural synchrony, copulation, allopreening responsiveness and total aggression. We operationally defined mates as more compatible when they showed greater proximity, synchrony, allopreening responsiveness and copulation frequencies and lower frequencies of aggression. Using a captive, seminatural population, we tested the following three interrelated hypotheses: (1) pairs with greater behavioural compatibility before breeding should have greater reproductive success; (2) among pairs that laid fertile eggs, pairs with greater behavioural compatibility should show better incubation coordination and (3) pairs with better incubation coordination should hatch more fertile eggs. Our results were consistent with these hypotheses. Pairs that laid eggs had significantly higher behavioural compatibility scores than did those that laid no eggs. Pairs with higher behavioural compatibility scores reared significantly more chicks to independence even after controlling for the effects of pair bond duration and parental age. Among pairs that laid fertile eggs, mates with higher behavioural compatibility scores had greater incubation coordination, measured by periods during which only one parent at a time attended the eggs, and greater hatching success of fertile eggs. Behavioural compatibility between mates in socially monogamous species may have reproductive consequences, and emergent behavioural features of pair relationships may have evolutionary significance.

Foraging opportunity and increased physical complexity both prevent and reduce psychogenic feather picking by young Amazon parrots

Abstract Although many authors have suggested that the quality of the cage environment contributes to the development and performance of psychogenic feather picking by parrots, there is little scientific evidence for this relationship. In chickens, there is an established relationship between absence of foraging opportunity and the performance of a similar behavior, feather pecking. Thus, we assessed whether providing environmental enrichments designed to facilitate foraging behaviors would prevent or reduce the development of feather picking behavior by parrots, as evidenced by superior feather condition. Two groups of eight parrots were parent-reared to weaning and then housed singly in either enriched or unenriched cages for 48 weeks. In the enriched condition, a unique combination of one foraging and one physical enrichment was presented to each parrot weekly. In both groups, feather condition was quantified using a 10-point scale. The provision of enrichments led to an improvement in feather condition over 48 weeks in the enriched group, while feather scores in the control group decreased significantly (repeated measures GLM: F 1,46 =5.59; P =0.022) during this same period, indicating that feather picking behavior had developed in this group. In the second part of this study, the control group was transferred to the enriched treatment for a period of 16 weeks. During this period re-feathering occurred and feather scores improved significantly, indicating that feather picking behavior had decreased (repeated measures GLM: F 1,53 =35.57, P

Plasma melatonin levels in Japanese quail exposed to dim light are determined by subjective interpretation of day and night, not light intensity.

Plasma melatonin levels were measured in male Japanese quail exposed to lighting schedules consisting of combinations of bright light (2000 or 1500 lx), darkness, or dim light (2 lx) or to constant dim light. Melatonin levels in dim light were dependent upon the relative intensity of accompanying phases, being significantly higher when dim light was subjective night than when it was subjective day. There was no significant melatonin rhythm in constant dim light, even on the first day of constant dim light exposure. Melatonin levels were intermediate when dim light was accompanied by both bright light and darkness. These results indicate that melatonin secretion in birds does not depend solely on light intensity. Furthermore, these results suggest that the avian circadian system may be more sensitive to environmental cues than its mammalian counterpart.

Neonatal handling of Amazon parrots alters the stress response and immune function

The influence of neonatal handling on behavior and immune function was assessed in Orange-winged Amazon parrots (Amazona amazonica). Chicks (n=11) were gently handled daily from 25 days of age until 38 days post-fledging, while control chicks (n=9) were not handled. At 10 days post-fledging ( approximately 66 days of age), chicks were given tests to evaluate tameness (e.g., willingness to perch on an offered finger). They were then restrained for 10 min, either by being held while perching (handled group) or, because they would not perch, by being restrained in a towel (nonhandled group). Serum corticosterone levels were measured and immune status was assessed by: the delayed-type hypersensitivity (DTH) response to phytohemagglutinin-P (PH-P) injection; the humoral response to a killed Newcastle disease virus (NDV) challenge; and heterophil:lymphocyte ratio (H:L). Handled chicks were tamer by all measures of tameness. DTH was greater in nonhandled chicks (P</=0.002), as were serum corticosterone levels (Wilcoxon, P</=0.05), while NDV antibody titers were possibly reduced (P</=0.09). H:L ratios did not differ. We conclude that handling conditioned the birds to be held in a manner that appeared not to be stressful. The greater DTH response of nonhandled chicks suggests that either their DTH response was enhanced by the acute stress of being restrained in a towel, and/or the DTH response of handled chicks was suppressed as a result of the repeated physiologic stress from handling during the neonatal period. In either event, handling produced marked differences in response to types of restraint that would be typically encountered in the husbandry of Amazons in captivity.

Brood patch innervation and its role in the onset of incubation in the Turkey hen

The source of innervation to the brood patch in turkey hens was determined by recording the electrophysiological activity of cutaneous nerves while manually stimulating various regions of the skin. The entire area of the brood patch was innervated by eight nerves, arising from thoracic vertebra 3 to synsacrothoracic vertebra 1. To determine whether afferent input from the brood patch influenced egg production or incubation behavior, hens were bilaterally denervated prior to photoinduced egg production. Denervated hens visited nests the same number of times, but stayed on the nest for less total time than controls by the fourth week of photostimulation. Serum prolactin levels rose in control hens but not in denervated hens. Egg production was maintained in the denervated hens but not in controls. None of the denervated hens displayed incubation behavior. This experiment supports the view that peripheral nervous input plays a role in the onset of incubation behavior.

Plasma LH and prolactin levels during the reproductive cycle of the Cockatiel (Nymphicus hollandicus)

Plasma-luteinizing hormone (LH) and prolactin (Prl) levels were determined using radioimmunoassay during two reproductive cycles in captive cockatiels (Nymphicus hollandicus)--an altricial species in which both parents share incubation and care of young. Birds were stimulated to breed by increasing daylength, light intensity, ambient temperature, and presenting nest boxes. LH levels were elevated during the time of nest inspection in females and peaked during egg laying. In contrast, LH levels were highest in males during nest inspection but were lower during egg laying. In both sexes, LH continued to decline during incubation and care of the young but rose in pairs laying a second clutch. Female and male Prl levels increased during egg laying, peaked during incubation, then declined to egg-laying levels during the nestling stage. Prl continued to decline during the fledgling stage and reached prelaying levels unless a second clutch was begun. In conclusion, in cockatiels, nest inspection and laying are characterized by high LH levels while high Prl levels occur during incubation and feeding of nestlings in both males and females.

Rhythm-dependent light induction of the c-fos gene in the Turkey hypothalamus

Day length (photoperiod) is a powerful synchroniser of seasonal changes in the reproductive neuroendocrine activity in temperate‐zone birds. When exposed to light during the photoinducible phase, reproductive neuroendocrine responses occur. However, the neuroendocrine systems involved in avian reproduction are poorly understood. We investigated the effect of light exposure at different circadian times upon the hypothalamus and components of the circadian system, using c‐fos mRNA expression, measured by in situ hybridisation, as an indicator of light‐induced neuronal activity. Levels of c‐fos mRNA in these areas were compared after turkey hens (on a daily 6‐h light period) had been exposed to a 30‐min period of light occurring at 8, 14, or 20 h after the onset of first light of the day (subjective dawn). Non‐photostimulated control birds were harvested at the same times. In birds, photostimulated within the photoinducibile phase (14 h), in contrast to before or after, c‐fos mRNA was significantly increased in the nucleus commissurae pallii (nCPa), nucleus premamillaris (PMM), eminentia mediana (ME), and organum vasculosum lamina terminalis (OVLT). Photostimulation increased c‐fos mRNA expression in the pineal gland, nucleus suprachiasmaticus, pars visualis (vSCN) and nucleus inferioris hypothalami compared to that of their corresponding nonphotostimulated controls. However, the magnitudes of the responses in these areas were similar irrespective of where in the dark period the pulses occurred. No c‐fos mRNA was induced in the nucleus infundibulari, in response to the 30‐min light period at any of the circadian times tested. The lack of c‐fos up‐regulation in the pineal gland and vSCN following photostimulation during the photoinducible phase lends credence to the hypothesis that these areas are not involved in the photic initiation of avian reproduction. On the other hand, c‐fos mRNA increases in the nCPa, ME, and OVLT support other studies showing that these areas are involved in the onset of reproductive behaviour initiated by long day lengths. The present study provides novel data showing that the PMM in the caudal hypothalamus is involved in the neuronally mediated, light‐induced initiation of reproductive activity in the turkey hen.

Identification of Dopamine, Gonadotrophin-Releasing Hormone-I, and Vasoactive Intestinal Peptide Neurones Activated by Electrical Stimulation to the Medial Preoptic Area of the Turkey Hypothalamus: A Potential Reproductive Neuroendocrine Circuit

The neural and neurochemical substrates regulating reproduction in birds remain vaguely defined. The findings that electrical stimulation in the medial preoptic area (ES/MPOA) or intracerebroventricular infusion of dopamine (DA) stimulated luteinising hormone (LH) and prolactin (PRL) release in female turkeys, led to the suggestion that ES/MPOA might help to clarify the DA circuitry regulating LH and PRL. We used c‐fos mRNA and tyrosine hydroxylase immunoreactivity as measured by double in situ hybridisation/immunocytochemistry (ISH/ICC) to determine which group/subgroup of DA neurones was activated following unilateral ES/MPOA. To establish that the reproductive neuroendocrine system was activated, double ISH/ICC was also conducted on c‐fos/gonadotrophin‐releasing hormone‐I (GnRH‐I) and c‐fos/vasoactive intestinal peptide (VIP). Changes in circulating LH and PRL were determined by radioimmunoassay. Unilateral ES/MPOA (100 µA, right side) of anaesthetised laying turkeys for 30 min increased circulating LH and PRL levels. It also induced c‐fos mRNA expression on the ipsilateral side by all GnRH‐I neurones within the septopreoptic region, implying that GnRH‐I neurones in this region share similar circuitry. VIP neurones within the nucleus infundibularis were the only VIP group to show c‐fos mRNA expression, suggesting their involvement in ES/MPOA induced PRL release. c‐fos mRNA expression was also observed in a subgroup of DA neurones in the nucleus mamillaris lateralis (ML). To our knowledge, the present study is the first to show that activation of DAergic cells in the ML is associated with the activation of GnRH‐I and VIP neurones and the release of LH and PRL. It is likely that ES/MPOA activated VIP/GnRH‐I neurones via activation of DA neurones in the ML, as this was the only DA subgroup that showed c‐fos mRNA expression.

Post-hatch oral estrogen exposure reduces oviduct and egg mass and alters nest-building behavior in adult zebra finches (Taeniopygia guttata)

In addition to well recognized effects on the zebra finch song system, we previously showed that post-hatch oral estrogen and xenoestrogen exposure disrupts reproduction by increasing eggshell breakage in females and decreasing fertility in males. Here we show that post-hatch exposure to estradiol benzoate (by oral gavage on days 5 to 11) at a 100 nmol EB per g body mass dose (EB100) also reduces adult oviduct mass. Further, EB100 and doses two orders of magnitude lower (EB10, EB1) reduce egg mass and length. Similar to the induction of song-control nuclei in females, dosing with EB10 and EB100 increased and masculinized another highly differentiated behavior: nest-building. Zebra finches orally exposed as chicks were observed during reproductive trials in communal breeding cages for 4 or 6weeks duration. EB100 males and females and EB10 males showed increased nest-building behaviors. Further, EB10 and EB100 birds had larger nests than canola oil-treated controls, and EB100 birds had faster rates of nest-building than controls, while EB1 birds had significantly slower rates of nest-building than controls. Additionally, EB100 males and females also showed an increased preference for a coarser male-typical nest-building material (jute) over a finer, female-typical material (wool), suggesting a masculinization of nest-building behavior at the higher doses. The change in zebra finch nest-building behavior induced by early EB exposure suggests that nest size and quality, in addition to egg mass and length, may provide new endpoints for assessing avian exposure to xeno- and phyto-estrogens in wild birds.

Posthatch Oral Estrogen Exposure Impairs Adult Reproductive Performance of Zebra Finch in a Sex-Specific Manner

We determined whether short-term, posthatch oral exposure to estradiol benzoate (EB) or the industrial surfactant octylphenol (OP) could impair the reproductive performance of zebra finches. If so, naturally occurring phytoestrogens and xenoestrogens might influence reproduction in wild populations. Chicks were given oral administration of 10 or 100 nmol EB per gram of body mass (earlier work showed the latter to be the minimum oral dose required to maximally masculinize female song nuclei) or an equimolar amount of OP daily from 5 through 11 days of age. Canola oil was used as a vehicle and control. Reproductive testing was done either in individual pair cages or in communal cages that permitted self-selection of mates, N = 10 pairs per group. Pairs consisted of EB-treated males and females, EB-treated males paired with canola-treated females, vice versa, and canola-treated males and females. Posthatch EB treatment produced sex-specific impairments in reproduction that, in some instances, were additive when both sexes were treated. Egg production was reduced and egg breakage was increased in 100 nmol/g EB-treated male and female pairs. The incidence of missing eggs was increased in 10 nmol/g EB-treated male and female pairs. Candled fertility was reduced in both groups containing 100 nmol/g EB-treated males. The number of hatched chicks was severely reduced in all EB-treated groups. No adverse effects of OP treatment were detected. These significant treatment effects (all P < 0.05) show that posthatch EB treatment profoundly disrupts the reproductive performance of zebra finches, suggesting that exposure to estrogens in the wild could impair the reproductive performance of wild populations.