Enhanced yolk testosterone influences behavioral phenotype independent of sex in Japanese quail chicks Coturnix japonica

Studies have demonstrated an effect of yolk testosterone levels on the physiology and behavior of nestling birds. In order to investigate this phenomenon experimentally in a precocial bird, we enhanced yolk testosterone, but within the physiological range, by injecting 50 ng testosterone in ethanol into Japanese quail Coturnix japonica eggs prior to incubation. The chicks hatching from these as well as from control eggs that had received the carrier-only or were left unmanipulated were subject to a number of behavioral tests from hatching to the age of 3 weeks. In addition, fecal samples were taken during a 90-min isolation period to determine a physiological response to a stressor. Experimental chicks performed a detour task faster and approached novel objects sooner than did the controls. Chicks from treated eggs took a longer time to start distress vocalizing and also produced less distress calls during open-field trials, took on average a larger number of trials for them to show tonic immobility and also excreted lower levels of corticosterone metabolites (BM) than did the controls. In response to a stressor, excreted BM was initially higher in the control chicks, as compared to the experimental birds. Induced behavioral effects were independent of sex with no sex treatment interactions found. In sum, experimentally enhanced testosterone levels in the eggs shifted individual behavioral phenotype towards "bold" or "proactive", irrespective of sex. We conclude that testosterone in the yolk influences the coping style of hatchlings and may be a potential means of maternal influence on offspring phenotype.     

Faecal corticosterone and immunoglobulin A in young adult rats

Quantitative analyses of relevant molecules in faeces may have potential as future non-invasive measures of stress. This study examined levels of faecal corticosterone and immunoglubulin A (IgA) in young adult rats and how these levels varied according to age, gender and time of day. Faecal samples were collected from 40 young adult rats (7 weeks old, n = 20 and 10 weeks old, n = 20) of both sexes from two time windows: day and night. The concentrations of corticosterone and IgA were measured by ELISAs following organic solvent extraction and aqueous extraction, respectively, of the molecules from faecal pellets. The production of faeces per time unit was higher in males than in females, and linear correlations were found between the faecal concentrations of corticosterone and IgA and total amounts of the respective molecules excreted in faeces per kg body weight per hour. In all further analyses the levels of the two molecules were calculated as amounts secreted per kg of body weight per hour. There was no gender difference between females and males in the production of corticosterone and IgA, but 7-week-old animals excreted significantly higher amounts of both molecules than did 10-week-old rats. The levels of IgA excreted by female rats were higher in the evening than in the morning, and male rats excreted higher concentrations of corticosterone in the morning than in the evening.     

Seasonal variation in growth of greater snow goose goslings: the role of food supply

Even though growth rate is an important fitness component, it is still controversial to what extent parent birds adjust the timing of offspring hatch to natural variations in food supply to maximize offspring growth. We studied the role of food availability in explaining inter- and intra-seasonal variation of growth rate in goslings of greater snow geese over 5 years. The peak of hatching generally coincided with the peak of food availability. However, early-hatched goslings usually grew faster than birds hatched at the peak, which in␣turn grew faster than late-hatched goslings, although this phenomenon was not observed in all years. There was considerable variation in growth rate among the five years, the smallest goslings produced in the best year (1991) being larger than the largest goslings of the poorest year (1994). We developed three indices of food availability, based on the cumulative availability of plant biomass and nitrogen content during the growth period, and showed that the cumulative exposure to nitrogen biomass explained up to 43% of variation (intra- and inter-annual) in body size just before fledging. In years with good feeding conditions, early-hatched goslings had access to more nitrogen during their growing period than those hatching on or after the peak and they grew faster. In years of lower food availability, early-hatched goslings had no detectable advantage over peak- or late-hatched birds for access to protein-rich food and no seasonal decline in growth rate was observed. These results confirm the critical role of food supply in the seasonal variation of growth rate in Arctic-nesting geese. Received: 27 June 1997 / Accepted: 9 October 1997     

Seasonal changes in testosterone and corticosterone levels in four social classes of a desert dwelling sociable rodent

Animals have to adjust their physiology to seasonal changes, in response to variation in food availability, social tactics and reproduction. I compared basal corticosterone and testosterone levels in free ranging striped mouse from a desert habitat, comparing between the sexes, breeding and philopatric non-breeding individuals, and between the breeding and the non-breeding season. I expected differences between breeders and non-breeders and between seasons with high and low food availability. Basal serum corticosterone was measured from 132 different individuals and serum testosterone from 176 different individuals of free living striped mice. Corticosterone and testosterone levels were independent of age, body weight and not influenced by carrying a transmitter. The levels of corticosterone and testosterone declined by approximately 50% from the breeding to the non-breeding season in breeding females as well as non-breeding males and females. In contrast, breeding males showed much lower corticosterone levels during the breeding season than all other classes, and were the only class that showed an increase of corticosterone from the breeding to the non-breeding season. As a result, breeding males had similar corticosterone levels as other social classes during the non-breeding season. During the breeding season, breeding males had much higher testosterone levels than other classes, which decreased significantly from the breeding to the non-breeding season. My results support the prediction that corticosterone decreases during periods of low food abundance. Variation in the pattern of hormonal secretion in striped mice might assist them to cope with seasonal changes in energy demand in a desert habitat.     

Individualities in a flock of free-roaming greylag geese: behavioral and physiological consistency over time and across situations

The concept of personality implies individual differences in behavior and physiology that show some degree of repeatability/consistency over time and across contexts. Most studies of animal personality, particularly studies of individuals' variation in physiological mechanisms, have been conducted on selected individuals in controlled conditions. We attempted to detect consistent behaviors as well as physiological patterns in greylag ganders (Anser anser) from a free-roaming flock living in semi-natural conditions. We tested 10 individuals repeatedly, in a handling trial, resembling tests for characterization of "temperaments" in captive animals. We recorded the behavior of the same 10 individuals during four situations in the socially intact flock: (1) a "low density feeding condition", (2) a "high density feeding condition", (3) a "low density post-feeding situation" and (4) while the geese rested. We collected fecal samples for determination of excreted immuno-reactive corticosterone (BM) and testosterone metabolites (TM) after handling trials, as well as the "low density feeding" and the "high density feeding" conditions. BM levels were very highly consistent over the repeats of handling trials, and the "low density feeding condition" and tended to be consistent over the first two repeats of the "high density feeding condition". Also, BM responses tended to be consistent across contexts. Despite seasonal variation, there tended to be inter-test consistency of TM, which pointed to some individual differences in TM as well. Aggressiveness turned out to be a highly repeatable trait, which was consistent across social situations, and tended to correlate with an individual's resistance during handling trials. Also, "proximity to the female partner" and "sociability" - the average number of neighboring geese in a close distance while resting - were consistent. We conclude that aggressiveness, "affiliative tendencies" and levels of excreted corticosterone and testosterone metabolites may be crucial factors of personality in geese.     

Hormonal Correlates of Dominance and Starvation-induced Aggression in Chicks of the Blue-footed Booby

In the blue-footed booby (Sula nebouxii), the first-hatched chick aggressively dominates its sibling and sometimes kills it when food is in short supply. To investigate the endocrine correlates of dominance-subordinance and hunger-induced agonism, we deprived 15–20-d-old single-chick and two-chick broods of food during 48 h by taping chicks' necks to prevent ingestion of parentally provided food (a protocol used previously and known to elicit escalated sibling fighting). We monitored weight and levels of circulating testosterone, oestradiol and corticosterone in deprived and normally fed broods comprising singletons, seniors (= dominants) and juniors (= subordinates), and observed behaviour to verify that aggression increased in deprived two-chick broods. During the 2 d of fasting, experimental chicks lost on average 6% of their baseline weight. After normal feeding was reinstated, seniors and singletons recovered normal (control) weight, but juniors remained significantly lighter than controls. No testosterone was detected in any nestling, but baseline corticosterone level was 109% higher in juniors than in seniors or singletons, implying that elevated corticosterone in juniors is a consequence of social subordination and may facilitate submissive behaviour. Although there was evidence that aggression of seniors increased under food deprivation, the increase was not accompanied by the rise in levels of testosterone expected under the ‘Challenge hypothesis’ (Wingfield et al. 1990). This result implies that this hypothesis probably does not apply to booby nestlings in the context of starvation-induced aggression. During the 2 d deprivation period, corticosterone levels of experimental chicks increased significantly, then declined to baseline levels a day after tapes were removed. The increase was probably due to the combined effects of starvation and frustration. We suggest that corticosterone could alter responsiveness of nestling blue-footed boobies to external stimuli, resulting in more aggression by dominants and greater readiness of subordinates to submit.     

Behavioural and hormonal effects of social isolation and neophobia in a gregarious bird species, the European starling (Sturnus vulgaris)

Separating gregarious individuals from their group members often results in behavioural and physiological changes, like increased levels of corticosterone. Testosterone and corticosterone, in particular, have been implicated in the response of mammals to novelty. Data in birds are, however, rare. The presence or absence of group members may also influence an individual's response to novel stimuli. We assessed the behaviour and hormonal response of European starlings (Sturnus vulgaris) to a novel object in two different situations and seasons: each starling was tested when separated and when in contact with its group members in May/June (breeding season) and again in September/October (non-breeding season). Starlings are gregarious throughout the year, but as foraging flocks are small during the breeding season and large during the non-breeding season, we assumed that non-breeding starlings would be more affected by social isolation. Overall, starlings had higher levels of corticosterone, lost more body mass, and were more active when they were separated from their group. Isolated individuals, however, did not show a greater neophobic response than individuals in the presence of their group members in either season. Circulating levels of testosterone and corticosterone were higher after a test with novel object than after a test with only the familiar feeding dish in both sexes and seasons. However, control tests for handling effects confirmed only the increase in testosterone. Our study shows that social isolation is stressful for unrelated and unpaired members of a wild flocking bird species and demonstrates that novelty can lead to a rise in testosterone in birds.     

Experimental Food Restriction Reveals Individual Differences in Corticosterone Reaction Norms with No Oxidative Costs

Highly plastic endocrine traits are thought to play a central role in allowing organisms to respond rapidly to environmental change. Yet, not all individuals display the same degree of plasticity in these traits, and the costs of this individual variation in plasticity are unknown. We studied individual differences in corticosterone levels under varying conditions to test whether there are consistent individual differences in (1) baseline corticosterone levels; (2) plasticity in the hormonal response to an ecologically relevant stressor (food restriction); and (3) whether individual differences in plasticity are related to fitness costs, as estimated by oxidative stress levels. We took 25 wild-caught house sparrows into captivity and assigned them to repeated food restricted and control treatments (60% and 110% of their daily food intake), such that each individual experienced both food restricted and control diets twice. We found significant individual variation in baseline corticosterone levels and stress responsiveness, even after controlling for changes in body mass. However, these individual differences in hormonal responsiveness were not related to measures of oxidative stress. These results have implications for how corticosterone levels may evolve in natural populations and raise questions about what we can conclude from phenotypic correlations between hormone levels and fitness measures.     

The effects of variable foraging conditions on common murre (Uria aalge) corticosterone concentrations and parental provisioning

This study investigated how total corticosterone concentrations, chick-feeding rates, and adult body mass changed with food availability from 1998 to 2000 in the same individually marked common murres (Uria aalge). Capelin, the main prey species, arrived inshore by the onset of murre chick hatching in 1998 and 1999 (prey match years); whereas in 2000, hatching began approximately 1 week before the capelin arrived inshore to spawn (prey mismatch year). Serum corticosterone concentrations were higher in the same individuals in the prey mismatch year than they were in either of the match years. Birds sampled before peak capelin spawning in the mismatch year had higher corticosterone levels than murres sampled after peak spawning. Murres with higher corticosterone levels had higher chick-feeding rates and less mass loss in the mismatch year (compared to the match year 1999) than birds with lower levels. Corticosterone levels did not differ between birds that had not foraged for at least 12 h (brooded chick overnight) and those that had, suggesting that short-term food deprivation did not affect corticosterone concentrations. Taken together, these findings suggest that the difference between years reflects a baseline shift in corticosterone levels, particularly in the high-quality birds that were able to increase both corticosterone concentrations and foraging effort.     

Comparing plasma and faecal measures of steroid hormones in Adelie penguins Pygoscelis adeliae

Physiological measurements of both stress and sex hormones are often used to estimate the consequences of natural or human-induced change in ecological studies of various animals. Different methods of hormone measurement exist, potentially explaining variation in results across studies; methods should be cross-validated to ensure that they correlate. We directly compared faecal and plasma hormone measurements for the first time in a wild free-living species, the Adelie penguin (Pygoscelis adeliae). Blood and faecal samples were simultaneously collected from individual penguins for comparison and assayed for testosterone and corticosterone (or their metabolites). Sex differences and variability within each measure, and correlation of values across measures were compared. For both hormones, plasma samples showed greater variation than faecal samples. Males had higher mean corticosterone concentrations than females, but the difference was only statistically significant in faecal samples. Plasma testosterone, but not faecal testosterone, was significantly higher in males than females. Correlation between sample types was poor overall, and weaker in females than in males, perhaps because measures from plasma represent hormones that are both free and bound to globulins, whereas measures from faeces represent only the free portion. Faecal samples also represent a cumulative measure of hormones over time, as opposed to a plasma ‘snapshot’ concentration. Our data indicate that faecal sampling appears more suitable for assessing baseline hormone concentrations, whilst plasma sampling may best define immediate responses to environmental events. Consequently, future studies should ensure that they select the most appropriate matrix and method of hormone measurement to answer their research questions.     

Experimental assessment of problem solving by <Emphasis Type="Italic">Milvago chimango </Emphasis>(Aves: Falconiformes)

We report a preliminary assessment of problem solving as an estimate of behavioural innovation and learning ability of a generalist and abundant raptor, Milvago chimango, under controlled conditions in aviaries. Experimental tests consisted in presentation of a Plexiglas box with four lids leading to isolated pieces of meat. We recorded time to first contact with the box and time from this first contact to gaining access to the pieces of meat. We recorded the number of attempts to open the box and reach the first portion of meat, and the total number of lids opened by each individual during five successive daily sessions. We found that individuals of M. chimango quickly approached and made contact with the Plexiglas box, and responded successfully to the novel feeding problem of reaching the food inside. In our study, performance of individuals was enhanced after solution of the novel task for the first time, as indicated by the progressively reduced time taken to access the food. Further, some individuals gained access to an increasing number of sections of the Plexiglas box during subsequent sessions, suggesting significant learning ability. Our results indicated M. chimango has a remarkable ability to obtain food in a novel situation, an observation that agrees with anecdotal reports of opening of receptacles to obtain food in urban environments. The results support the idea of behavioural plasticity in this species.     

Effect of testosterone loading on the kinetic of faecal testosterone excretion in mallards

Intestinal passage time of coloured fodder and testosterone turnover were examined by faecal steroid analysis in mallards in the reproductive and postrefractory period. In the latter, the discharge of coloured fodder began 36 minutes after ingestion in males, and 56 minutes in females. During reproduction the discharge began 93 minutes and 112 minutes after ingestion in males and females, respectively. Total passage time was similar in the reproductive and postrefractory period in both sexes. After intraperitoneal testosterone injection, faecal samples were collected for 8 hours and testosterone levels were measured using RIA. In the postrefractory period, 1-2 hours after testosterone loading a strong increase of faecal testosterone content developed in males, meanwhile a slighter testosterone peak appeared in females. During reproduction testosterone excretion began 1.5-2 hours after injection in both sexes but in females its increase was smaller. The duration of response to testosterone loading was 5 hours in both periods and both sexes. Intensive excretion after T loading appeared earlier in males than in females, but total passage time finished at the same time: 5 hours after loading. The character of testosterone excretion was corresponding to the passage of fodder-chimus-faeces in the reproductive and postrefractory period in both sexes.     

Energy metabolism, testosterone and corticosterone in white-crowned sparrows

The influence of the steroid hormones testosterone and corticosterone on energy metabolism and activity of birds is largely enigmatic. We measured resting metabolic rate during night and day in 12 long-term castrated and 12 intact male white-crowned sparrows (Zonotrichia leucophrys gambelii) under short-day (8:16 SD), long-day (20:4 LD), LD+testosterone implant and LD−testosterone implant conditions. Each male was sequentially measured under all four conditions. Photostimulation increased testosterone, resting metabolic rate, food intake, hopping activity and body mass in castrates and intact males. Surprisingly, testosterone levels and metabolic rates did not differ between intact and castrated males. Testosterone implantation increased activity and food intake, but decreased body mass and resting metabolic rate in both groups. Removing testosterone implants reversed the effects on resting metabolic rate, activity and food intake. Corticosterone levels, measured immediately at the end of metabolism measurements, showed birds were not stressed. Corticosterone had no apparent relationship with resting metabolic rate and there was no interaction between corticosterone and testosterone. Overall, positive changes in testosterone levels resulted in a decrease of resting metabolic rate. We speculate that testosterone increases activity, and birds compensate for increased activity metabolism by reducing resting metabolic rate. Accepted: 18 July 1999     

Effects of caecectomy in the young adult female rat on digestibility of food offered ad libitum and in restricted amounts.

Caecectomized and sham-operated rats were fed a laboratory chow ad libitum and the effects of caecectomy on the digestibility of the food were studied. Compared with sham-operated controls, caecectomized rats showed a decrease in apparent digestibiltiy of organic matter from 77.8 to 73.0%, of crude protein from 83.0 to 79.4%, and of 'carbohydrate' from 74.6 to 69.0%. However, faecal water content increased from 41.6 to 54.8%. 51Cr-labelled EDTA was excreted faster in the faeces after caecectomy. The colon partly adapted to the loss of caecal mucosa by increased length and thus mucosal surface area. In a second concurrent experiment the effect of caecectomy on the apparent digestibility of food during food restriction was studied. Six caecectomized rats, comparable in all respects to those used in the first study, were fed the laboratory chow ad libitum for 3 weeks. They were then fed submaintenance amounts of food to achieve body weight losses of 40--50% and to maintain these low weights for 4 weeks. Finally, they were again fed ad libitum for 3 weeks. During the period of restriction the apparent digestibility of organic matter increased from 72.7 to 75.4%. This was largely due to the increased apparent digestibility of crude protein which rose from 78.4 to 81.9%. Digestibility coefficients returned to control values immediately upon refeeding ad libitum.     

Individual birds advance offspring hatching in response to increased temperature after the start of laying

In seasonally reproducing organisms, timing reproduction to match food availability is key to individual fitness. Ambient temperature functions as an important cue for the timing of the food peak in temperate-zone birds. After laying start, individual birds may still improve synchrony between offspring hatching and food availability by adjusting the onset of incubation to most up-to-date cues about the development of the food source. However, it is unknown whether individuals respond to changes in temperature after the onset of laying, and whether individuals adjust incubation onset independent of clutch size. Here, I show in free-living blue tits (Cyanistes caeruleus) that experimental heating of nestboxes in the laying phase resulted in increased duration of nocturnal incubation bouts prior to clutch completion, leading to earlier hatching of eggs and increased hatching asynchrony. Experimental heating did not affect the number of laying gaps, egg volume and clutch size, nor were any carry-over effects on offspring detected. These results are best explained as a response to increased temperature acting as a cue for an advanced food-peak, rather than a relief of energetic constraints, because improved energetic conditions would not favour more hatching asynchrony. Other benefits cannot be excluded, since increased laying-phase incubation under warmer conditions may also help maintain egg viability. This study is the first to show that temperature has a causal effect on the time between clutch completion and hatching of the first offspring, indicating that behavioural adjustment to climate change can continue after laying start.     

外源性皮质酮对雄性根田鼠交配行为的作用

本文研究了皮质酮对雄性根田鼠交配行为的作用。实验个体分别注射0.10 ug/ g体重、0. 60 ug / g体重和1.00 ug/ g体重剂量的皮质酮,1 h后测定每只雄性根田鼠的交配行为。结果显示,3 个处理组动物具有射精能力的个体比率及其爬跨、抽动和射精潜伏期与对照组动物相比均无显著差异,爬跨和抽动频次也无显著变化。不同处理组个体的血浆睾酮含量无显著差异。因此,皮质酮没有影响雄性根田鼠的交配行为和性激素的分泌。该结果提示,哺乳动物在急性应激条件下所分泌的皮质酮可能不参与对性行为的调控。     

Effects of testosterone and corticosterone on immunocompetence in the zebra finch

The original immunocompetence handicap hypothesis (ICHH) suggested that testosterone has a handicapping effect in males by both promoting the development of sexual signals and suppressing immune function. A modified version, the stress-linked ICHH, has recently proposed that testosterone is immunosuppressive indirectly by increasing production of corticosterone. To test both the original and stress-mediated versions of the ICHH, we implanted male zebra finches taken from lines selected for divergent maximum stress-induced levels of corticosterone (high, low and control) with either empty or testosterone-filled implants. Their humoral and cell-mediated immune responses were then assessed by challenge with diphtheria:tetanus vaccine and phytohemagglutinin respectively. We found no effect of the hormone manipulations on either PHA or tetanus antibody responses, but found a significant interaction between titers of both testosterone and corticosterone on diphtheria secondary antibody response; antibody response was greatest in individuals with high levels of both hormones. There was also a significant interactive effect between testosterone treatment group and corticosterone titer on body mass; the body mass of males in the elevated testosterone treatment group decreased with increasing corticosterone titer. These results suggest that, contrary to the assumption of the stress-mediated version of the ICHH, high plasma levels of corticosterone are not immunosuppressive, but are in fact immuno-enhancing in the presence of high levels of plasma testosterone. Equally, the central assumption of the ICHH that testosterone is obligately immunosuppressive is also not supported. The same individuals with the highest levels of both hormones and consequently the most robust antibody response also possessed the lowest body mass.     

The adrenocortical stress-response of Black-legged Kittiwake chicks in relation to dietary restrictions

In this study we examined hormonal responses of Black-legged Kittiwake (Rissatridactyla) chicks to experimental variations in energy content and nutritional quality (low or high lipid to protein ratio, LPR) of their food. Starting at the age of 10 days, chicks were fed either high or low LPR fish at 30, 50, 70 and 100% of ad libitum energy intake. After 20 days of treatment, chicks were exposed to a standardized acute handling and restraint stress protocol, where a baseline sample was taken immediately after taking a chick from the nest, and three additional blood samples were taken at intervals up to 50 min. Testosterone and corticosterone titres in plasma were measured via radioimmunoassay. We found that baseline testosterone levels were not significantly affected by the experimental treatments. Food-restricted chicks had elevated baseline and acute stress-induced levels of corticosterone compared to chicks fed ad libitum. An elevation of circulating levels of corticosterone in energetically stressed individuals was further magnified by low nutritional quality of food. Baseline and acute stress-induced corticosterone levels of chicks were negatively correlated with their fat reserves. We conclude that the physiological condition of Black-legged Kittiwake chicks can be assessed reliably by measuring circulating levels of corticosterone. We discuss short- and long-term effects of elevated corticosterone secretion in food-stressed nest-bound chicks. Accepted: 13 April 1999     

Resource partitioning in sympatric arctic-breeding geese: summer habitat use, spatial and dietary overlap of Barnacle and Pink-footed Geese in Svalbard

The spatial, habitat and dietary overlap of two breeding goose species was studied in Sassendalen, Svalbard, in summer 2003 based on abundance within 500 × 500‐m grid squares and faecal diet analyses during pre‐breeding, nesting and post‐hatching periods. More than half of all Pink‐footed Geese Anser brachyrhynchus occurred in the absence of Barnacle Geese Branta leucopsis during nesting and post‐hatching periods compared to c. 20% when concentrated by pre‐breeding snow cover. In contrast, only 5% of Barnacle Geese were observed in the absence of Pink‐footed Geese pre‐breeding, 15% during nesting, and 35% post‐hatching. Among six defined habitat types, Barnacle Geese resorted more to ‘upland’ habitats during pre‐breeding and nesting and to lowland lakes post‐hatching when compared to Pink‐footed Geese. Although Pink‐footed Geese showed less change in seasonal habitat preference, many shifted to the river valley bottom post‐hatching, giving access to open water (predator avoidance) and lush green vegetation (foraging for goslings). The smallest extent of distributional overlap between the two species occurred post‐hatching, but each species was also highly restricted by snow cover during pre‐nesting. The greatest extent of overlap in distribution and diet occurred during incubation, when large dietary variation between different breeding valleys reflected local food availability around nests (probably a result of nest‐site preference rather than food selection per se). Whether this means that increased interactions within and between the two goose species with future increases in local density are most likely to be manifest at this stage of the summer is impossible to determine without knowledge of available food resources and manipulative experiments. More detailed investigations of the effects of foraging by both species on plant structure, quality and community composition are necessary to predict likely outcomes of future changes in population densities of both species.     

Does Climate or Habitat Affect the Frequency of Cooperative Brood Rearing in Canada Geese?

Canada geese ( Branta canadensis ) are unusual because individuals use either of two different brood-rearing behaviors: cooperative broods (two or more merged broods attended by more than three parents) or two-parent families. We tested whether cooperative broods form in response to habitat or climatic conditions by examining variation in cooperative brood frequencies among Canada geese nesting in Connecticut from 1982 to 1996. Percent of goslings raised in cooperative broods ranged from 0 to 100% at a given site in different years, but the pattern of annual variation was different at each site. The sites were in close proximity to each other and had similar climates; thus, the differences in annual variation among sites was not likely to be a response to climatic conditions. Cooperative brood frequencies also varied among sites in each individual year, but sites with the most gang brooding in one year often had the least the next. Such would not be expected if gang brooding occurred in response to non-ephemeral habitat characteristics. Sites where gang brooding occurred and where it did not have similar food resources and predation risks. These findings failed to support the hypotheses that gang broods form in response to food competition or predation. Sites where gang broods occurred had more parent geese and more goslings than sites where they did not occur. Furthermore, the proportion of goslings raised in gang broods was correlated with the number of goslings and parents at the site. Our results support the hypothesis that gang broods form from the inadvertent mixing of goslings. This single factor, however, was not sufficient to account for all of the observed variation in gang brooding frequencies.