From the field to the lab: Muriqui endocrinology from a collaborative perspective.

The northern muriqui (Brachyteles hypoxanthus) is a critically endangered species endemic to the Atlantic Forest of southeastern Brazil. Long-term observational studies of wild muriquis have provided many insights into the behavioral ecology and life history of this species. However, nearly everything that is currently known about the northern muriqui's behavioral endocrinology has come from combining our respective expertise in noninvasive field and laboratory research. Here, we reflect on the history of our collaboration, focusing on major challenges, key scientific findings, and factors that contributed to its success. Challenges included insuring the reliable collection of frequent fecal samples from a large enough number of known individuals over extended periods of time, preserving the steroids in the field and transporting them, developing and validating the fecal steroid assays, and interpreting the hormonal profiles within the behavioral and ecological contexts of the study subjects. Major findings included our thorough description of the fecal progesterone and estradiol profiles associated with muriqui ovarian cycling and gestation, the seasonal resumption of cycling, its onset during puberty of dispersing females, and the differences between fertile and infertile cycles. We also documented the relationship between fecal cortisol and testosterone in sexually active males across breeding and nonbreeding seasons, and sex differences in cortisol levels across the mating and conception seasons. We attribute the success of our collaboration to a number of factors including our mutual appreciation for one another's high standards for ethics, data quality, and data interpretation.

Artificial insemination in black-handed spider monkey (Ateles geoffroyi).

Artificial insemination (AI) was performed in spider monkeys; these primates are vulnerable to extinction and usually do not reproduce spontaneously in captivity. Uterine cycles were followed by daily assessment of vaginal cytology, and corroborated a posteriori by concentrations of 17-beta estradiol and progesterone, measured by radioimmunoassay (RIA), in fecal samples collected once daily. Five females between 13 to 27 years old were inseminated intravaginally (with fresh semen) twice each during the periovulatory phase (Days 9-12 of the menstrual cycle; Day 0, first day of menstrual bleeding), from September to the first 3 weeks of November (most fertile months). Transcervical AI was not useful in this primate because the liquid portion of the semen completely solidified instead of liquefying as in other primates. Pregnancies were apparently achieved in 5 of 14 attempts. One female became pregnant after the first round of inseminations, delivered a healthy infant, was inseminated and got pregnant again (subsequently aborted). One female aborted, apparently due to an intramural uterine leiomyoma. Another two females stopped menstruating for a few months, then restarted menstruating (these females may have been pregnant and aborted). In conclusion, in spider monkeys: (1) captivity-induced stress did not inhibit reproduction; (2) fecal steroid hormones were useful to assess cyclicity; (3) the semen coagulum, which apparently is a tightly packed and large reservoir of spermatozoa, must not be discarded but used in AI; (4) old female spider monkeys did not have cessation of reproductive function.

Seasonal variations of fecal progesterone and 17beta-estradiol in captive female black-handed spider monkeys (Ateles geoffroyi).

A number of studies in free-ranging and captive spider monkeys (Ateles spp.) have shown that this genus is able to reproduce throughout the entire year. Nonetheless, it is still controversial whether births, and therefore conceptions, tend to be more frequent during certain seasons. In the present study, we monitored changes in fecal 17beta-estradiol and progesterone for approximately 1 years in five female black-handed spider monkeys (Ateles geoffroyi) kept in captivity in Mexico City. The objective was to determine whether hormone concentrations and menstrual cycles of summer and autumn accounted for a greater chance of conception than those of winter and spring, consistent with birth patterns previously reported. We collected fecal samples from the five monkeys almost daily for 1 year (March 2004 to February 2005) and used radioimmunoassay of fecal extracts to determine concentrations of 17beta-estradiol and progesterone. Concurrently, menstrual cycle phases were determined by cytological evaluation of vaginal swabs. Periovulatory 17beta-estradiol concentrations were significantly higher in autumn than in winter, spring or summer. Moreover, as evidenced by progesterone peaks, most of the summer-autumn menstrual cycles were ovulatory; in contrast, most of the winter and spring cycles were anovulatory. In conclusion, our data supported the notion that, although not a strictly seasonal reproducer, the black-handed spider monkey is more likely to conceive at the end of the rainy season and throughout autumn.

Germinación de semillas de Ficus insipida (Moraceae) defecadas por tucanes (Ramphastos sulfuratus) y monos araña (Ateles geoffroyi) / Germination of Ficus insipida (Moraceae) seeds from toucan (Ramphastos sulfuratus) and spider monkey (Ateles geoffroyi) feces

To test the null hypothesis that two vertebrate fructivores, toucans (Ramphastos sulfuratus) and spider monkeys (Ateles geoffroyi), are equally specialized in germinating Ficus insipida seeds after these have passed through their digestive tracts, we fed fruits to captive individuals. We extracted seeds from feces and placed them on filter paper in petri dishes under controled light, temperature and humidity. Control seeds had not passed through a digestive tract. We found that a greater proportion of seeds from A. geofroyi (65%) germinated (R. sulfuratus: 4%). The germinative value was also greater in seeds from monkey feces (rate=13.76; toucan 0.046; control group 0.172). If, despite individual variability of seeds and dispersers, future studies continue to show that A. geoffroyi favors germination more (maybe because of a longer digestion time), this would indicate that diet specialization is not necessarily related to dispersal efficiency.

Con el objetivo de evaluar y comparar el efecto que tiene el paso de las semillas del higo Ficus insipida Willd (Moraceae), por el tracto digestivo de dos especies de vertebrados frugívoros, se ofrecieron frutos a individuos cautivos de tucán (Ramphastos sulfuratus: Ramphastidae) y de mono araña (Ateles geoffroyi: Cebidae), en la hipótesis de que siendo ambos frugívoros especialistas deberían favorecer de manera similar a la germinación de semillas de esta especie arbórea. De las heces se separaron las semillas y se pusieron a germinar en cajas petri en condiciones controladas de luz, temperatura y humedad, con una muestra de semillas testigo que no pasaron por el tracto digestivo de ningún animal. Se calculó la latencia, la capacidad y velocidad de geminación, y el valor germinativo de las semillas de cada grupo experimental. El 65% de las semillas de frutos de F. insipida consumidos por A. geofroyi germinaron con éxito, mientras que de las obtenidas de las heces de tucán, apenas el 4% logró germinar. El valor germinativo fue mayor en las semillas extraídas de excretas de mono (índice=13.76), en comparación con las de tucán (0.046) y las del grupo testigo (0.172). A. geoffroyi fue la especie que favoreció más la germinación de semillas de F. insipida, siendo probable que esta condición se relacione con el mayor tiempo de digestión de este primate. Deberá considerarse ambién la dificultad de establecer siempre concordancia con estos resultados, debido a la gran variabilidad en la capacidad germinativa de las semillas de individuos arbóreos, a la diferencia entre las cosechas y periodos de fructificación, así como en la madurez y estado fisiológico de los animales utilizados en las pruebas. Se concluye que la especialización alimentaria no necesariamente se relaciona con la efectividad de la calidad de la dispersión.