Oligonucleotide fingerprinting of free-ranging and captive rhesus macaques from Cayo Santiago: paternity assignment and comparison of heterozygosity.

Multilocus DNA fingerprinting with oligonucleotide probes (GTG)5, (GATA)4, and (CA)8 was applied in order to determine paternity in one birth cohort (15 infants) of social group (S) from the free-ranging colony of rhesus macaques (Macaca mulatta) on Cayo Santiago. While sires could be identified in 11 cases, all males tested (N = 19) could be excluded from paternity for the remaining four infants. Data revealed marked discrepancies between actual paternity and paternity as inferred from the observation of copulation behavior. Thus, a dominant social rank does not appear to be strongly associated with reproductive success. Furthermore, alternative reproductive strategies were found to yield comparable net benefits in reproduction. A second group of animals (M) was translocated from Cayo Santiago to the Sabana Seca Field Station in 1984. They have continuously resided together in a large outdoor enclosure since then. Here paternity assessment was seriously impeded by a reduced number of discriminating bands, i.e. offspring bands which were unequivocally derived from the sires. This was initially held to be indicative of a smaller degree of heterozygosity in Group M, and was attributed to inbreeding due to a lack of male immigration or extra-group fertilizations. However, a comparison of the DNA fingerprint patterns obtained in Group S and Group M lends only partial support to this idea.

Dominance rank, cortisol concentrations, and reproductive maturation in male rhesus macaques.

Among adolescent male rhesus macaques, Macaca mulatta, the highest ranking individual within a cohort has higher testosterone concentrations at a younger age, earlier in the mating season, and for a longer duration than his lower ranking conspecifics. We sought to determine whether such a rank-related pattern of reproductive maturation could be a function of differences in glucocorticoid levels. A 2-yr longitudinal study of a cohort of adolescent males living in a heterosexual group in a one acre outdoor enclosure revealed no differences in cortisol concentrations between high and low status males. Cortisol was not inversely correlated with testosterone in either adolescent or adult males. Young pubescent males had increases in cortisol levels coincident with maturation, while older adolescent males had cortisol concentrations comparable to those of adult males. Low ranking males tended to have more variable cortisol concentrations across time. We conclude that cortisol concentrations are not a function of dominance status and that the timing of reproductive maturation in male rhesus macaques is independent of cortisol concentrations.

Future research on Cayo Santiago-derived group M rhesus monkeys at Sabana Seca: the socioendocrinology of male reproductive development.

Group M is an intact social group transferred from Cayo Santiago to Sabana Seca in 1984 and will be used for research integrating biomedical and behavioral investigations. The initial research planned for Group M will assess how social factors mediate the interactions between hormones and behavior as they affect male reproductive maturation. Preliminary data indicate that social status can accelerate endocrine maturation in males and that high ranking males may have a head start in initiating their reproductive careers. Elucidating the socioendocrinology of male reproductive development will complement the many studies that have been undertaken on the sociobiology of male rhesus macaques at Cayo Santiago.

Plasma ascorbic acid concentrations in a population of rhesus monkeys (Macaca mulatta).

Although monkeys frequently are used as animal models for ascorbic acid studies whose results are extrapolated to humans, little information is available on the normal levels of this vitamin in large populations of animals classified by sex, age, or physiologic state such as pregnancy or lactation. The purpose of this report is to provide these values and compare them to the same parameters in humans, pointing out similar and dissimilar trends. Plasma samples were obtained from a troop of 167 rhesus monkeys (Macaca mulatta) and analyzed for ascorbic acid by using the 2,4-dinitrophenyl hydrazine method. Results obtained for ascorbic acid concentrations in plasma showed no differences between sexes. A significant (P< 0.0001) lowering effect was observed in aging versus young animals. Pregnant and nonpregnant females had similar ascorbate values, and lactating monkeys had slightly elevated levels. We conclude that rhesus monkeys and humans exhibit some of the same characteristics of ascorbic acid metabolism, such as an age-related decrease in ascorbate and the maintenance of these levels during lactation. However, a difference between species was noted with gender. Women maintain higher ascorbate concentrations than do men, whereas no differences in concentrations of this vitamin were observed between female and male monkeys.

Consequences of group fission for the patterns of relatedness among rhesus macaques.

When mammalian social groups exceed their optimal size, they often tend to split. In view of the potential evolutionary benefits, it should be more advantageous for animals to stay with kin, rather than nonkin, during such fission events. In the present study, the spontaneous fission of two social groups, R and S, of rhesus macaques living on Cayo Santiago, Puerto Rico, provided the opportunity to compare the kinship structure of the corresponding parent and daughter groups, using information on both maternal and paternal relatedness. In both instances, maternal half-siblings and pairs of animals from the same family were significantly more prevalent in the fission products than in the parent group. During the split of group R, significantly more paternal half-siblings stayed in the remnants of the parent group than joined the seceding group. Our findings are compatible with previous behavioural studies demonstrating that female primates bias their social behaviour more to maternal than to paternal kin, but that both types of half-siblings prefer each other more than unrelated animals. It remains to be clarified by future research, however, whether the observed co-segregation of paternal half-sibs in our study reflects active choice or is a by-product of the group-specific kin structures, prior to fission.

Dominance rank and reproductive maturation in male rhesus macaques (Macaca mulatta).

The timing of reproductive maturation in females has been linked to differences in dominance status and exposure to males, but less information is available regarding the effect of dominance rank and exposure to females on the pace of reproductive maturation in males. A two-year study of a cohort of nine male rhesus macaques was undertaken to document potential social factors influencing variation in rates of reproductive maturation. The males lived in a large heterosexual group housed in a 0.3 ha outdoor enclosure. Every four weeks the focal subjects were isolated from the group to collect morphometric measurements and blood samples. The results revealed that dominance rank was not significantly correlated with average testosterone concentrations during the mating season. However, the highest ranking adolescent male had higher testosterone concentrations at a younger age, earlier in the mating season, and for a longer duration than did the lowest ranking male. Relative testicular weight was significantly correlated with dominance rank during the premating season, but not the postmating season, among adolescent males. The highest ranking adolescent male also engaged in more affiliative and sexual activity with nonkin, sexually receptive females than did lower ranking conspecifics. These patterns of development provide evidence that high dominance status accelerated reproductive maturation in male rhesus macaques.

Estradiol concentrations, fat deposits, and reproductive strategies in male rhesus macaques.

Squirrel monkeys (Saimiri spp.) are thought to undergo a unique pattern of seasonal weight change among the Primates that reflects a fatted male phenomenon. But many male mammals, including rhesus macaques (Macaca mulatta), undergo circannual weight changes concurrent with mating season activity. Four adult male rhesus macaques living in a heterosexual social group in an outdoor enclosure were studied over a 2-year period in order to ascertain potential mechanisms underlying seasonal weight changes. Alterations in weight were significantly correlated with changes in abdominal fat levels and in body mass index. Neither testosterone nor estradiol fluctuations were correlated with weight changes, but estradiol levels were significantly correlated with body mass index. The annual profile of changes in abdominal fat level reflected the annual profile of variation in circulating estradiol levels. The fatted male phenomenon appears to characterize circannual weight fluctuations in rhesus macaques. Seasonal weight changes resulting from fat deposition are suggested to provide a substrate for adipose tissue aromatization that yields elevated estradiol levels and enables males to forego feeding and concentrate their energy budgets on activities directly related to mate acquisition and retention.

Annual birth patterns of savanna baboons (Papio cynocephalus anubis) over a ten-year period at Gilgil, Kenya.

Most cercopithecines reproduce on a seasonal basis, but the proximate mechanisms influencing birth periodicity are often unclear. We analyze 10 years of data from the Gilgil Baboon Project (Kenya) in order to examine the relationship between annual birth patterns and rainfall. Savanna baboons at Gilgil copulated in all months of the year, and births did not occur on a seasonal basis. Annual rainfall patterns showed no association with annual birth patterns, but the chances of conception were significantly greater following the end of the long rainy season than at other times of the year. Nonseasonal reproduction is a general characteristic of savanna baboons (Papio cynocephalus subsp.) throughout Africa. The extreme dietary diversity of baboons enables them to utilize a wide variety of resources and facultatively manipulate the timing of different stages in the reproductive cycle in accordance with resource availability. We predict that nonseasonality of reproduction will be more likely to occur among species with a large dietary diversity than among species with a more restricted diet.

Socioendocrine and morphological correlates of paternity in rhesus macaques (Macaca mulatta).

When females mate with several males, problems arise in identifying sire and in determining factors contributing to differential male reproductive success. Three potential primary correlates of differential reproduction in males include fighting ability, sperm competition, and body condition. We collected a variety of socioendocrine and morphological measurements from sexually mature rhesus macaques to determine corollaries of paternity. We studied a troop of about 150 rhesus macaques living in a 0.3 ha corral and identified the sires of 70% of infants using multilocus DNA fingerprints. Eight of 21 males sired offspring, and dominant males were more successful than subordinate males. Neither canine size nor age influenced the probability of siring offspring. Male reproductive success was primarily an outcome of the number of females mated with, which was associated with an ensemble of traits including high dominance rank, large body size, relatively voluminous testicles and good body condition. Testes size was significantly larger in sires than in non-sires, but among sires the number of progeny produced was not correlated with testicle size. Sires began the mating season with more body fat than non-sires, but the energetic costs of mating resulted in a 50% reduction in abdominal skinfold thickness during the mating season. We conclude that social status exerts a major impact on paternity by affecting the number of females mated with, that male quality is a critical factor modulating paternity, and that male feeding strategies have a direct influence on variation in male reproductive success.

Genetic determination of paternity and variation in male reproductive success in two populations of rhesus macaques.

The fundamental framework for uncovering factors affecting the evolution of social behavior rests upon analyses of variation in reproductive success. In species where females mate with multiple males, paternity is invisible in the absence of genetic data. We determined paternity in two populations of rhesus macaques, Macaca mulatta, using both single locus and multilocus techniques. One troop, Group R, is one of four troops living on a 15 ha island (Cayo Santiago) off the coast of Puerto Rico, while the other troop, Group M, was translocated from Cayo Santiago to the Sabana Seca Field Station (Puerto Rico) in 1984. About a dozen human-derived short tandem repeat (STR) markers have been found to be polymorphic in the study of populations and provide the initial paternity determination. Final evaluation of paternity is then confirmed by multilocus DNA fingerprinting using synthetic oligonucleotide probes. Body condition, age, and dominance rank have an impact on male progeny production, while canine size does not. We suggest that nonagonistic competition in the form of sperm competition and endurance rivalry will modulate male reproductive success. A large body size among males provides them with an advantage in both sperm competition and endurance rivalry. Comparison of the two populations indicated that demographic, social, ecological, and morphological factors interact to regulate variation in reproductive success among male nonhuman primates.

Behavioral masculinization is independent of genital masculinization in prenatally androgenized female rhesus macaques.

Genetic female fetuses were exposed transplacentally to testosterone propionate injected into their mothers either early (Days 40 through 64) or late (Days 115 through 139) in gestation. Early and late androgenized females (EAFs and LAFs, respectively) were raised with normal males and females that served as criteria for evaluating degree of behavioral masculinization induced by the prenatal androgen. EAFs were genitally virilized and LAFs were not. Males and untreated females differed reliably on five behavioral measures: males showed more mother-mounting, more peer-mounting, more rough play with peers, a preference for initiating play with male partners, and less grooming of mothers. Neither type of prenatally androgenized female showed masculinization of all five types of behavior. Compared with females, EAFs showed more mother-mounting, more peer-mounting, less mother-grooming, did not differ from females in rough play, and did not manifest a preference for male partners. LAFs, like females, groomed but did not mount their mothers, and did not show a preference for male partners; but unlike females they showed elevated rough play and mounting with peers. EAFs showed a statistically significant delay in puberty onset (menarche), but LAFs did not. Mothers inspected genitalia of their offspring more often if they were males than if they were females. Mothers of EAFs inspected their offspring's genitalia as often as mothers of males, but mothers of LAFs did not. No aspect of maternal behavior was associated with either the amount or kind of masculine behavior shown toward peers. We interpret the results to mean that genital virilization is independent of, and largely irrelevant to, the expression of those behavioral traits that characterize the juvenile male social role. Moreover, the individual behavior traits that are components of the juvenile male role are independently regulated by the organizing actions of androgen and have separable critical periods. Of the two major traits, mounting peers and rough play with peers, the latter has a greater requirement for androgenic stimulation late in prenatal life.

The effects of isosexual rearing on adult sexual behavior in captive male rhesus macaques.

The sexual competence of males reared only with other males (isosexually reared) was compared to the sexual competence of males reared in mixed-sex peer groups (heterosexually reared). All subjects were pair-tested with the same 11 females. Isosexually reared males tended to be less sexually active than heterosexually reared males, but the differences were not pronounced. Heterosexually reared males took less time to ejaculate than isosexually reared males, but their ejaculatory frequencies were comparable. Among one peer group of isosexually reared males, their level of sexual competence conformed with their dominance ranks when immature. These results suggest that growing up with female peers is not a prerequisite for the expression of adult sexual behavior among male rhesus macaques and that adult male sexual behavior may be influenced by relative dominance status when immature.

Does intermale mounting function as a dominance demonstration in rhesus monkeys?

The relationship between dominance and intermale mounting was analyzed in two troops of captive rhesus monkeys (Macaca mulatta). The data did not support the assumption that mounting among males functions as a dominance demonstration as described in the literature because mounting commonly (63% of 65 dyads) occurred in a bidirectional fashion, and mounting not only was performed by dominant males but also was shown with regularity (36% of 521 cases) by subordinate males. Intermale mounting was often (37% of 521 cases) associated with amicable interactions--commonly preceding them--but it was only rarely (2%) associated with dominance-subordination behaviors. It was concluded that intermale mounting may serve as a socially cohesive behavior in rhesus monkeys by promoting nonagonistic contact.

Increased microsatellite variability in Macaca mulatta compared to humans due to a large scale deletion/insertion event during primate evolution.

Human (GATA)n microsatellites D12S66 and D12S67 could be successfully amplified by polymerase chain reaction (PCR) in various species of apes and monkeys. In 86 unrelated animals of the most intensively studied species Macaca mulatta we demonstrated five alleles at "D12D66" differing in size in increments of 4 bp (159-175 bp), whereas 17 alleles were observed at locus "D12S67". The alleles of the latter locus are distributed in two separate groups with no alleles of intermediate size. Six alleles were found between 108-128 bp and 11 alleles between 181-249 bp. Mendelian inheritance of the codominant allele was proven by family studies. Sequencing of the "D12S67" locus revealed that the shorter alleles are characterized by a single perfect (GATA)n stretch whereas the longer alleles consist of two blocks of (GATA)n repeats separated by an intervening sequence of 9 bp. The composite structure of the longer alleles closely resembles that of the 12 human D12S67 alleles (229-273 bp). The enormous species variation in the fragment size range, with the smallest allele found in Macaca mulatta (108 bp) and the largest (364 bp) in Gorilla gorilla gorilla strongly indicates that D12s67 has been subjected to recurrent mutations over the course of primate evolution including a large deletion and/or insertion event.

Primigravidity, body weight, and costs of rearing first offspring in rhesus macaques.

Body size is associated with menarche and ovarian function, but the relationship to first conception is rarely examined. We conducted a longitudinal investigation of rhesus macaques, Macaca mulatta, to determine the effect of differences in body weight on both the age at first conception and survivorship of first progeny. Young females who became pregnant weighed significantly more than peers who remained barren, with weight changes for females who successfully raised offspring no different than those for females who did not produce offspring. Infant mortality among primiparae was not significantly greater than that among multiparae, although nearly twice as high. First-born males tended to have lower survivorship than first-born female offspring. We suggest that reproductive costs encountered by primiparous females are more likely to be modulated by immature neuroendocrine function than by inexperience, small body size, or infant suckling patterns. We conclude that body size influences probability of first conception, socioendocrine factors mediate the likelihood of infant survival, and primiparous production of male progeny seems to exert a greater reproductive cost than does production of female progeny.

Subcutaneous rabies vaccination of pigtail macaques.

Nonhuman primates housed in outside cages in rabies enzootic areas should be vaccinated against rabies because of the risk of exposure to rabid wildlife. This article reports that vaccination with an inactivated rabies vaccine for domestic animals induced levels of neutralizing antibodies against rabies in pigtail macaques (Macaca nemestrina) without causing adverse reactions.

Paternal relatedness and age proximity regulate social relationships among adult female rhesus macaques.

Kin selection promotes the evolution of social behavior that increases the survival and reproductive success of close relatives. Among primates, maternal kinship frequently coincides with a higher frequency of grooming and agonistic aiding, but the extent to which paternal kinship influences adult female social relationships has not yet been investigated. Here, we examine the effect of both maternal and paternal kinship, as well as age proximity, on affiliative interactions among semifree-ranging adult female rhesus macaques, Macaca mulatta. Kinship was assessed by using both microsatellites and DNA-fingerprinting. Our study confirms that the closest affiliative relationships characterize maternal half-sisters. We provide evidence that adult females are significantly more affiliative with paternal half-sisters than with nonkin. Furthermore, paternal kin discrimination was more pronounced among peers than among nonpeers, indicating that age proximity has an additional regulatory effect on affiliative interactions. We propose that kin discrimination among cercopithecine primates emerges from ontogenetic processes that involve phenotype matching based on shared behavioral traits, such as inherited personality profiles, rather than physiological or physical characteristics.

A non-invasive technique for the control of intestinal parasites in rhesus macaques (Macaca mulatta).

Control of intestinal parasites is a major concern in maintaining the health and well-being of captive primates. This study established that percutaneous application of ivermectin was as effective as subcutaneous administration for the control of nematodes in captive rhesus macaques. The method is less intrusive than the standard subcutaneous or oral delivery routes and did not adversely affect the health of any of the study subjects.

Paternity assessment in rhesus macaques (Macaca mulatta): multilocus DNA fingerprinting and PCR marker typing.

Establishing kinship relations in primates using modern molecular genetic techniques has enhanced the ability to scrutinize a number of fundamental biological issues. We screened 51 human short tandem repeats (STRs) for cross-species PCR amplification in rhesus macaques (Macaca mulatta) and identified 11 polymorphic loci with heterozygosity rates of at least 0.6. These markers were used for paternity testing in three social groups (M, R, and S) of rhesus macaques from Cayo Santiago, Puerto Rico. Several consecutive birth cohorts were analyzed in which approximately 200 males were tested for paternity against more than 100 mother/ infant pairs. Despite a combined exclusion rate of more than 99.9% in all three groups, some cases could not be solved unequivocally with the STR markers and additional testing of the MHC-associated DQB1 polymorphism. A final decision became possible through multilocus DNA fingerprinting with one or more of the oligonucleotide probes (GATA)4, (CA)8, and (CAC)5. Paternity assessment by multilocus DNA analysis with probe (CAC)5 alone was found to have limitations in rhesus macaques as regards the number of potential sires which might be involved in a given case. Multilocus DNA fingerprinting requires large amounts of DNA, and the ensuing autoradiographic patterns present difficulties in comparisons across gels and even within the same gel across remote lanes. Computer-assisted image analysis was incapable of eliminating this problem. Therefore, a dual approach to DNA typing has been adopted, using STR markers to reduce the number of potential sires to a level where all remaining candidates can be tested by multilocus DNA fingerprinting on a single gel, preferably in lanes adjacent to the mother/infant pair.

Increased reproductive success of MHC class II heterozygous males among free-ranging rhesus macaques.

Gene conversion and balancing selection have been invoked to explain the ubiquitous diversity of the antigen-presenting proteins encoded in the vertebrate major histocompatibility complex (MHC). In the present study, direct evidence for over-dominant selection promoting MHC diversity in primates is provided by the observation that, in a large free-ranging population of rhesus macaques, males heterozygous at MHC class II locus Mamu-DQB1 sired significantly more offspring than homozygotes (the male-specific selection coefficient s equals 0.34). This heterozygote advantage appeared to be independent of the actual male Mamu-DQB1 genotype. No similar effect emerged for a captive group of monkeys of similar genetic background but under veterinary care.