No abstract

The Greater Rhea (Rhea americana) is a nearthreatened species. Wild populations are affected by human activities, such as illegal hunting, egg harvesting and conversion of natural habitats to croplands. An indicator of disturbances is the increase of the glucocorticoid corticosterone, a stress hormone that helps to cope with lifethreatening situations. Here, we evaluate and characterize adrenocortical function in the Greater Rhea by validating the use of a radioimmunoassay (RIA) to assess fecal glucocorticoid metabolite concentrations, and by comparing the time course of plasma corticosterone and fecal immunoreactive glucocorticoid metabolite excretion. An adrenocorticotropic hormone (ACTH) challenge was performed and serial blood and fecal (cecal and rectal) samples were collected and analyzed by a corticosterone RIA.High-performance liquid chromatography (HPLC) was employed to characterize fecal immunoreactive glucocorticoid metabolites. HPLC analysis revealed the presence of two immunoreactive glucocorticoid metabolites in rectal and cecal Rhea feces. Greater Rheas responded to the ACTH challenge with a 30-fold increase in plasma corticosterone concentration 2 h post-injection, and a 30-to 40-fold increase in concentrations of glucocorticoid metabolites from rectal feces 4-6 h post-injection. In cecal feces, ACTH challenge produced an 11-fold increase. Therefore, collection of rectal feces would be more appropriated to ensure detection of minor increases in adrenocortical activity in this species. The adrenocortical response to ACTH found in Rhea was higher than what is usually observed in other birds and might be associated with their flightless condition and the use of running as an antipredator strategy.

The effect of transport stress on blood corticosterone levels in captive Greater Rheas was investigated. Twelve adult individuals (7 males; 5 females) were loaded in pairs inside wooden crates and transported along a paved road for 30 min. Blood samples were taken before the individuals were introduced into the crate (baseline value) and immediately after they were unloaded (30 min after capture). To assess whether corticosterone levels were affected by the blood sampling procedure per se, another 6 (nontransport) control birds (3 males; 3 females) were also captured and sampled at the same times as their transported counterparts. Plasma corticosterone concentrations were measured using a commercially available corticosterone (125)I radio-immunoassay kit. Baseline corticosterone levels were similar in the control and transported birds (9.0 ± 1.6 and 10.4 ± 0.8 ng/mL, respectively). Transportation induced a highly significant (P < 0.001), more than 40-fold increase in the corticosterone levels (433.6 ± 35.4 ng/mL) that was about 5 times higher (P < 0.001) than in their nontransported counterparts (88.4 ± 14.8 ng/mL). The present findings suggest that Greater Rhea is a species highly sensitive to stressful manipulations. Both blood sampling and transportation induced highly significant adrenocortical responses. Considering that transportation is one of the unavoidable common practices in the management of Greater Rheas and, as shown in the present study, that it induces a significant 40-fold corticosterone stress response, efforts should focus on helping to generate management transport standards for optimization of the welfare of this ratite.

Stress in freilebenden Nandupopulationen Rhea americana: Effekte landwirtschaftlicher Aktivita ¨ten auf saisonale Glukokortikoidmetabolitspiegel im Kot In den letzten Jahren haben freilebende Populationen des Nandus (Rhea americana) drastisch abgenommen, hauptsa ¨chlich wegen der Umwandlung von Gru ¨nland in Ackerland infolge intensiver, spezialisierter landwirtschaftlicher Methoden. In dieser Studie testen wir mo ¨gliche Stresseffekte A. Le `che (&)

No abstract

Seasonal rhythm in sex hormones has been extensively studied in birds, as well as its relationship with the type of mating system. The Greater Rhea (Rhea americana), a South American ratite species, reproduces seasonally and has a complex mating system: female-defense polygyny and sequential polyandry. The present study aimed at analyzing the endocrine basis of reproduction in this species and its relationship with its mating system. We used HPLC and electrochemiluminescence techniques to identify and measure plasma testosterone and estradiol levels. Annual oscillations in sex hormones, testosterone and estradiol, in adult males and females were observed. Lower levels of these hormones were exhibited during the non reproductive season (February to July), whereas their maximum values were reached in September for males and November-December for females. These fluctuations reflect the seasonal changes in gonadal function. By contrast, no significant sex hormones oscillations were observed in juvenile males and females (negative control of seasonal changes). Greater rheas maintain high testosterone and estradiol levels throughout the reproductive period. The high testosterone levels during incubation and chick rearing did not inhibit parental behavior in males, which appears not to conform to the “Challenge Hypothesis”. In females, the high estradiol levels throughout the reproductive season would be needed to sustain their long egg-laying period.

The Greater Rhea (Rhea americana) is an endemic ratite to South America, whose wild populations have undergone a remarkable decrease due to habitat degradation and fragmentation by the expansion of the agricultural frontier, poaching and predation by dogs. Anthropogenic perturbations in wild environments, as well as the management in captivity, can generate different stress responses in this species, thus, the monitoring of adrenocortical and behavioral activities are considered primary assessment tools with both conservation and welfare implications. In this review we analyze and integrate the different measurements of glucocorticoids (in plasma, feces, and yolk) carried out in different captive and wild populations, taking into account the diverse predictable and unpredictable conditions to which the Greater Rhea responds in each of those environments. In addition, the translocation of this bird is presented as an application of stress physiology in field ecology for conservation purposes, in which we evaluated how this species responds when it is released into a novel environment. Our results indicate that this ratite has a striking high sensitivity of the hypothalamic-pituitary-adrenal (HPA) axis compared to that of other bird species and shows a wide variety of adrenocortical responses depending on the environment in which it lives. This suggests that its HPA axis has a phenotypic plasticity that enables the rhea to cope with the environmental challenges. In this sense, we propose that one of the routes of this plasticity could be mediated by the maternal transfer of steroid hormones to the egg. Finally, we discuss the importance of integrating the monitoring of the adrenocortical response along with the environmental variables that define the life history of the species, in management and conservation programs ex-situ and in situ.

Animal transport is an indispensable practice in species that need to be moved for management or commercial purposes. However, transport may have negative effects on individuals' welfare. The aims of the present work were to determine if the behavioral responses of adult Greater Rheas (Rhea americana) bred in captivity are sensitive to short-term transport and if males and females differ in their posttransport behavioral activity and recovery. Eight males and 8 females were placed in individual pens and allowed 6 d to habituate (d 1 to 6) before transport procedure. On the transport day (d 7), half of the birds (4 males and 4 females) were randomly assigned to a transport group that was captured and handled to be placed into the crates, exposed to a 30-min transport stressor, and immediately returned to their pens. Four transports with 1 different male and female each time were performed. The other half remained undisturbed and were used as controls. Behavior of all individuals was video-recorded during habituation days, after transport on d 7, and on the 2 following days (d 8 and 9) to evaluate pre- and posttransport behavioral activity for 2 h per day. No significant behavioral changes were observed during the last 2 d of the habituation period (d 5 and 6), suggesting that Greater Rheas were adapted to the housing conditions before transport. After transportation, several behaviors were affected: transported males and females showed null resting, transported females also showed reduced preening and increased vigilance (P < 0.05), whereas transported males showed increased drinking (P < 0.05) compared with their respective control groups. The results suggest that behavioral responses of captive-bred Greater Rheas are sensitive to short-term transport (which includes handling) and that males and females differ in their posttransport behavioral activity, recovering their overall basal levels on the third day posttransportation.