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Analysis of glucocorticoid profiles serves as a valuable, multi-faceted tool for insight into the behavior and physiology of wild populations. Recently, the measurement of fecal glucocorticoid metabolites (FCMs) has exploded in popularity due to its compatibility with noninvasive techniques and remote environments A critical first step is to perform a biological validation to ensure that the assay accurately reflect changes in FCM levels. We use an enzyme immunoassay (EIA) to perform a biological validation on samples collected from two males and six females in a wild population of Colobus vellerosus in response to three naturally occurring potential stressors. We also describe the FCM response pattern in the week following parturition in three females and examine the influence of sex, reproductive state, and time of day on the concentrations of baseline samples collected daily from 13 adult individuals over a period of four months. We validated the assay: FCM levels increase in response to natural stressors with a two-day lag. In the two days surrounding parturition, FMC levels increased. Baseline concentrations were affected by collection time and female reproductive state, with lactating females having lower concentrations than pregnant or cycling females. Thus, we successfully carried out the first validation and characterization of FCMs in a wild African colobine. This will serve as an essential foundation for future studies of C. vellerosus and similar wild primates whose objective is to investigate the role glucocorticoids play in responses to social and ecological challenges.

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Numerous studies have examined the effects of anthropogenic endocrine disrupting compounds; however, very little is known about the effects of naturally occurring plant-produced estrogenic compounds (i.e., phytoestrogens) on vertebrates. To examine the seasonal pattern of phytoestrogen consumption and its relationship to hormone levels (407 fecal samples analyzed for estradiol and cortisol) and social behavior (aggression, mating, and grooming) in a primate, we conducted an 11-month field study of red colobus (Procolobus rufomitratus) in Kibale National Park, Uganda. The percent of diet from estrogenic plants averaged 10.7% (n=45 weeks; range: 0.7-32.4%). Red colobus fed more heavily on estrogenic Millettia dura young leaves during weeks of higher rainfall, and the consumption of this estrogenic item was positively correlated to both their fecal estradiol and cortisol levels. Social behaviors were related to estradiol and cortisol levels, as well as the consumption of estrogenic plants and rainfall. The more the red colobus consumed estrogenic plants the higher their rates of aggression and copulation and the lower their time spent grooming. Our results suggest that the consumption of estrogenic plants has important implications for primate health and fitness through interactions with the endocrine system and changes in hormone levels and social behaviors.

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Ruminants are characterized by an efficient particle-sorting mechanism in the forestomach (FRST) followed by selective rechewing of large food particles. For the nonruminating foregut fermenter pygmy hippo it was demonstrated that large particles are excreted as fast as, or faster than, the small particles. The same has been suggested for other nonruminating foregut fermenters. We determined the mean retention time of fluids and different-sized particles in six red kangaroos (Macropus rufus), seven collared peccaries (Pecari tajacu) and three colobine monkeys (Colobus angolensis, C. polykomos, Trachypithecus johnii). We fed Co-EDTA as fluid and mordanted fiber as particle markers (Cr, Ce). Mean (+ or - SD) total tract retention time for fluids, small and large particles was 14 + or - 2, 29 + or - 10 and 30 + or - 9 hr in red kangaroos, 26 + or - 2, 34 + or - 5 and 32 + or - 3 hr in collared peccaries and 57 + or - 17, 55 + or - 19 and 54 + or - 19 hr in colobine monkeys, respectively. Large and small particles were excreted simultaneously in all species. There was no difference in the excretion of fluids and particles in the colobine monkeys, in contrast to the other foregut fermenters. In the nonprimate, nonruminant foregut fermenters, the difference in the excretion of fluids and small particles decreases with increasing food intake. On the contrary, ruminants keep this differential excretion constant at different intake levels. This may be a prerequisite for the sorting of particles in their FRST and enable them to achieve higher food intake rates. The functional significance of differential excretion of fluids and particles from the FRST requires further investigations.

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If stress and disease impose fitness costs, and if those costs vary as a function of group size, then stress and disease should exert selection pressures on group size. We assessed the relationships between group size, stress, and parasite infections across nine groups of red colobus monkeys (Procolobus rufomitratus) in Kibale National Park, Uganda. We used fecal cortisol as a measure of physiological stress and examined fecal samples to assess the prevalence and intensity of gastrointestinal helminth infections. We also examined the effect of behaviors that could potentially reduce parasite transmission (e.g., increasing group spread and reducing social interactions). We found that cortisol was not significantly related to group size, but parasite prevalence was negatively related to group size and group spread. The observed increase in group spread could have reduced the rate of parasite transmission in larger groups; however, it is not clear whether this was a density-dependent behavioral counter-strategy to infection or a response to food competition that also reduced parasite transmission. The results do not support the suggestion that gastrointestinal parasitism or stress directly imposed group-size-related fitness costs, and we cannot conclude that they are among the mechanisms limiting group size in red colobus monkeys.

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Similar morphological or physiological changes occurring in multiple evolutionary lineages are not uncommon. Such parallel changes are believed to be adaptive, because a complex character is unlikely to originate more than once by chance. However, the occurrence of adaptive parallel amino acid substitutions is debated. Here I propose four requirements for establishing adaptive parallel evolution at the protein sequence level and use these criteria to demonstrate such a case. I report that the gene encoding pancreatic ribonuclease was duplicated independently in Asian and African leaf-eating monkeys. Statistical analyses of DNA sequences, functional assays of reconstructed ancestral proteins and site-directed mutagenesis show that the new genes acquired enhanced digestive efficiencies through parallel amino acid replacements driven by darwinian selection. They also lost a non-digestive function independently, under a relaxed selective constraint. These results demonstrate that despite the overall stochasticity, even molecular evolution has a certain degree of repeatability and predictability under the pressures of natural selection.

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