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Megan E. Brown

Publications and source records attributed to Megan E. Brown.

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Egg production and endocrine profiles of female whooping cranes (Grus americana) maintained ex situ are improved in naturalized enclosures

Whooping cranes ( Grus americana ) are naturally seasonal breeders and rely on wetland habitats throughout the annual cycle. However, captive cranes are commonly housed in dry outdoor pens, which may lack key environmental stimuli that in turn compromise reproduction. Our study sought to first, assess seasonal patterns of reproductive hormones in successful versus non-successful bird pairs and second, determine endocrine responses to alterations in enclosure environments. Fecal samples were collected from eight crane pairs year-round for 3 consecutive years, once a week during non-breeding season and three times a week during the breeding season. During Year 1, all pairs were housed in traditional dry pens, whereas during January of Year 2 all pairs were moved to either new dry pens (control, n = 4) or ponded pens (wetland, n = 4) and remained in the same pen until the end of the study. Fecal samples were assessed for gonadal (year-round) and adrenal (breeding season only) hormones. Estrogen and progestagen metabolite concentrations were low during non-breeding season in all females. However, as birds transition into a breeding season, gonadal hormone concentrations significantly increased ( p < 0.001) in laying females but not in non-laying individuals. Androgen metabolite concentrations during breeding seasons were higher ( p < 0.001) in males paired with non-laying females, with no variations observed during non-breeding months and within birds of the same reproductive output. We observed significant effect of enclosure environment on ovarian function of female whooping cranes. Specifically, mean estrogen metabolite concentrations increased after birds were moved from dry pen to wetland enclosures (Year 1: 349.1 ± 83.4 ng/g feces; Year 2: 382.7 ± 82.9 ng/g feces; Year 3: 556.5 ± 85.4 ng/g feces, p = 0.008 and 0.019 res p ectively,), whereas those of the control females remained constant. Further, estrogen concentration assessed during breeding season of Year 3 in females housed in wetland pens was higher than birds housed in a dry pen (556.5 ± 85.4 vs 311.7 ± 85.12 ng/g feces; p = 0.019). The number of eggs laid increased in three of the four pairs housed in the wetland pens, while there was no change in egg production in control birds (9 vs. 2 combined number of eggs produced by all pairs in each respective group in Year 3). Finally, moving birds to an enclosure that mimic natural environment did not impact androgen or glucocorticoid excretion. The findings demonstrate that differences in gonadal hormone production between laying and non-laying whooping crane females exist primarily during the breeding season, and that a more natural environment can have a positive influence on ovarian function in female whooping cranes.

Theriogenology Wild

Low estradiol production of non-laying whooping cranes (Grus americana) is associated with the failure of small follicles to enter follicular hierarchy

For endangered species managed ex situ , production of offspring is a key factor to ensure healthy and self-sustaining populations. However, current breeding goals for the whooping crane ( Grus americana ) are impeded by poor reproduction. Our study sought to better understand mechanisms regulating ovarian function in ex situ managed whooping cranes and the regulatory function of the hypothalamic-pituitary-gonadal (HPG) axis in relation to follicle formation and egg laying. To characterize hormonal regulation of follicular development and ovulation, we collected weekly blood samples from six female whooping cranes during two breeding seasons, for a total of 11 reproductive cycles. The plasma samples were assessed for follicle stimulating hormone, luteinizing hormone, estradiol, and progesterone and the yolk precursors vitellogenin and very low-density lipoprotein. Ultrasonographic examination of the ovary was conducted at the time of blood collection. Preovulatory follicles (>12 mm) were present in laying cycles (n = 6) but absent in non-laying cycles (n = 5). The patterns of plasma hormone and yolk precursor concentrations corresponded to the stage of follicle development. Specifically, gonadotropin and yolk precursors concentrations increased as follicles transitioned from the non-yolky to yolky stage but did not increase further as the follicle advanced to preovulatory and ovulatory stages. Estrogen and progesterone concentrations increased as follicle size increased and reached peak concentrations (P < 0.05) when follicles developed to ovulatory and preovulatory stages, respectively. While overall mean circulating gonadotropin, progesterone, and yolk precursor concentrations did not differ for laying versus non-laying cycles, mean plasma estradiol in laying cycles was significantly higher than that in non-laying cycles. In summary, the findings suggested that disruption of mechanisms regulating follicle recruitment is likely responsible for the oviposition failure of the captive female whooping crane.

General and Comparative Endocrinology

Comparison of fecal glucocorticoid metabolite concentrations in hand‐ versus parent‐reared whooping cranes (Grus americana)

Endangered whooping cranes ( Grus americana ) have been produced in captivity for reintroduction programs since the 1980s, using techniques such as artificial insemination, multiple clutching, and captive‐rearing to speed recovery efforts. Chicks are often hand‐reared (HR) by caretakers in crane costumes, socialized into groups and released together, unlike parent‐reared (PR) cranes that are raised individually by a male/female crane pair and released singly. HR cranes historically exhibit greater morbidity rates during development than PR cranes, involving musculoskeletal and respiratory system disease, among others. We hypothesized that HR crane chicks exhibit a higher baseline fecal glucocorticoid metabolite (FGM) concentrations during the development compared with PR chicks. Fecal samples were collected between 15 and 70 days of age from HR ( n = 15) and PR ( n = 8) chicks to test for differences in FGM concentrations using a radioimmunoassay technique following ethanol extraction for steroids. Linear mixed model analysis suggests increasing age of the chick was associated with an increase in FGM ( p < .001). Analysis also supported the interaction between rearing strategy and sex of the crane chick ( p < .01). Female PR chicks had greater FGM concentrations than all other groups (PR male, p < .01; HR female, p < .001; and HR male, p < .001). This result suggests that there may be an effect of rearing strategy on stress physiology of whooping crane chicks, especially among females. Further research is needed to investigate whether the FGM concentrations are reflective of true differences in stress physiology of young cranes and whether this may impact health and conservation success.zo

Zoo Biology

Female gonadal hormones and reproductive behaviors as key determinants of successful reproductive output of breeding whooping cranes ( Grus americana )

Reproductive success of endangered whooping cranes ( Grus americana ) maintained ex situ is poor. As part of an effort to identify potential causes of poor reproductive success in a captive colony, we used non-invasive endocrine monitoring to assess gonadal and adrenal steroids of bird pairs with various reproductive outcomes and evaluated the relationships of hormones and behaviors to reproductive performance. Overall, reproductively successful (i.e., egg laying) females had significantly higher mean estrogen levels but lower mean progestogen concentrations than did unsuccessful females. Other hormones, including glucocorticoids and androgens, were not significantly different between successful and unsuccessful individuals. Observations of specific behaviors such as unison calling, marching, and the number of copulation attempts, along with overall time spent performing reproductive behaviors, were significantly higher in successful pairs. Our findings indicate that overall reproductive performance of whooping crane pairs is linked to female gonadal hormone excretion and reproductive behaviors, but not to altered adrenal hormone production.

General and Comparative Endocrinology

Chromic and iron oxides as fecal markers to identify individual whooping cranes

The whooping crane ( Grus americana ) is listed as endangered under the IUCN Red List, the United States Endangered Species Act, and the Canadian Species at Risk Act (BirdLife International 2012, CWS and USFWS 2007). A major focus of recovery efforts for this endangered species is reintroduction to establish new populations (CWS and USFWS 2007). Captive populations are critical as a source of individuals for reintroduction efforts and also serve as insurance populations. Currently, there are a total of 157 whooping cranes held in captive breeding centers across North America, with the largest at the USGS Patuxent Wildlife Research Center (PWRC) in Laurel, Maryland. Birds produced in this facility are currently being released as part of efforts to establish the Eastern Migratory Population (EMP, Urbanek et al. 2005) and in an effort to establish a non-migratory population in Louisiana. In the past decade, PWRC has produced and released annually an average of 18 birds into the wild; however, reproductive performance of birds at this facility is lower than desired. PWRC had a 60% fertility rate for eggs laid from 2000 through 2010 (J. N. Chandler, personal communication, 2011). Furthermore, reproductive onset in this captive population appears to be delayed compared to wild populations. In wild populations, reproductive onset (production of sperm and eggs) normally occurs ~5 years of age in both males and females, ~2 years after initial pair formation occurs (Ellis et al., 1996), while some females in the EMP have laid eggs earlier than 5 years of age (Converse et al. 2011). However, PWRC females in some cases do not start to lay eggs until 7 years of age (Mirande et al. 1996). Currently, the PWRC population consists of a total of 74 whooping cranes, including 22 pairs. Six of these pairs (27%) are consistently infertile (i.e., no production of fertile eggs) and 3 other pairs (14%) have low fertility (30- 45% fertility in eggs laid), which is variable from year to year. Six pairs (27%) are recently formed and have not produced eggs, and so have unknown fertility. This leaves only 7 pairs (33%) which contribute maximally to PWRC’s chick production (J. N. Chandler, personal communication, 2011). Because of the challenges occurring within this captive colony, PWRC and Smithsonian National Zoo have initiated a joint research project to identify potential underlying causes of poor reproduction in captive whooping cranes.

Conference Paper