Metabolic Characterization of the Common Marmoset (Callithrix jacchus)

High-resolution metabolomics has created opportunity to integrate nutrition and metabolism into genetic studies to improve understanding of the diverse radiation of primate species. At present, however, there is very little information to help guide experimental design for study of wild populations. In a previous non-targeted metabolomics study of common marmosets (Callithrix jacchus), Rhesus macaques, humans, and four non-primate mammalian species, we found that essential amino acids (AA) and other central metabolites had interspecies variation similar to intraspecies variation while non-essential AA, environmental chemicals and catabolic waste products had greater interspecies variation. The present study was designed to test whether 55 plasma metabolites, including both nutritionally essential and non-essential metabolites and catabolic products, differ in concentration in common marmosets and humans. Significant differences were present for more than half of the metabolites analyzed and included AA, vitamins and central lipid metabolites, as well as for catabolic products of AA, nucleotides, energy metabolism and heme. Three environmental chemicals were present at low nanomolar concentrations but did not differ between species. Sex and age differences in marmosets were present for AA and nucleotide metabolism and warrant additional study. Overall, the results suggest that quantitative, targeted metabolomics can provide a useful complement to non-targeted metabolomics for studies of diet and environment interactions in primate evolution.     

Growth of the Mandible in the Common Marmoset (Callithrix jacchus)

The mandibles of 27 male and 25 female common marmosets (Callithrix jacchus) with an age range of 1 day to < 12 years have been studied using radiographs and osteometric techniques. The rate and magnitude of dimensional changes in the mandible of this nonhuman primate during the neonatal period, infancy, childhood, adolescence, and adulthood are described. Distance growth curves indicate that active growth of marmoset mandible ceases during the last 6 months of the third postnatal year.     

Characterization of Obese Phenotypes in a Small Nonhuman Primate,the Common Marmoset (Callithrix jacchus)

This report explores aspects of developing obesity in two captive populations of common marmosets (Callithrix jacchus), a small primate with a short lifespan that may be of value in modeling chronic aspects of obesity acquisition and its lifetime effects. Two populations were examined. In study 1, body composition, lipid parameters, and glucose metabolic parameters were measured in a population of 64 adult animals. Animals classified as obese (>80th percentile relative fat based on sex) displayed both dyslipidemia (higher triglyceride and very low–density lipoprotein (VLDL)) and altered glucose metabolism (higher fasting glucose and HbA_1c). Using operational definitions of atypical values for factors associated with metabolic syndrome in humans, five subjects (7.8%) had at least three atypical factors and five others had two atypical factors. A previously unreported finding in these normally sexually monomorphic primates was higher body weight, fat weights, and percent fat in females compared to males. In a second study, longitudinal weight data for a larger population (n = 210) were analyzed to evaluate the development of high weight animals. Differences in weights for animals that would exceed the 90th percentile in early adulthood were evident from infancy, with a 15% difference in weight between future‐large weight vs. their future‐normal weight litter mates as early as 4–6 months of age. The marmoset, therefore, demonstrates similar suites of obesity‐related alterations to those seen in other primates, including humans, suggesting that this species is worthy of consideration for obesity studies in which its fast maturity, high fertility, relatively short lifespan, and small size may be of advantage.     

Ontogeny of the Locomotor Activity Rhythm in the Common Marmoset (Callithrix jacchus)

The ontogeny of locomotor activity rhythm in the common marmoset was studied in three animals, observed during infant and juvenile stages, and in adult age. Animals were kept with their families in outdoor cages under natural temperature, humidity and light cycles. Data were collected from sunrise to sunset, three consecutive days a week, at 5 min intervals randomly distributed in the clock hours, and were recorded as the total of arbitrary cage sections traveled by the animal each interval. Animal 3 was observed continuously one day a week in order to detect less than 2 h period harmonic components. Animals' weekly data series were analyzed by Fourier analysis and Siegel's test to detect significant harmonic components. Results showed a main 24 h period com ponent for all records and a secondary 8 h period component in at least 34% of animal's week data series. The power content of the 24 h component showed an increase with age, reaching some stability near the 16th week of life suggesting that this component is mature at the end of the animal's infant stage. However, for the 8 h component an oscillating pattern was observed following the environmental annual temperature cycle; this may indicate a seasonal modulation of daily locomotor activity profiles.     

Ontogeny of the Locomotor Activity Rhythm in the Common Marmoset (Callithrix jacchus)

The ontogeny of locomotor activity rhythm in the common marmoset was studied in three animals, observed during infant and juvenile stages, and in adult age. Animals were kept with their families in outdoor cages under natural temperature, humidity and light cycles. Data were collected from sunrise to sunset, three consecutive days a week, at 5 min intervals randomly distributed in the clock hours, and were recorded as the total of arbitrary cage sections traveled by the animal each interval. Animal 3 was observed continuously one day a week in order to detect less than 2 h period harmonic components. Animals' weekly data series were analyzed by Fourier analysis and Siegel's test to detect significant harmonic components. Results showed a main 24 h period com ponent for all records and a secondary 8 h period component in at least 34% of animal's week data series. The power content of the 24 h component showed an increase with age, reaching some stability near the 16th week of life suggesting that this component is mature at the end of the animal's infant stage. However, for the 8 h component an oscillating pattern was observed following the environmental annual temperature cycle; this may indicate a seasonal modulation of daily locomotor activity profiles.     

Radiographic Evaluation of Appendicular Skeletal Maturation in the Common Marmoset (Callithrix jacchus)

The thoracic appendicular skeletal development of five common marmosets was monitored radiographically at weekly intervals from 1 day to 94 weeks of age and the times of appearance and fusion of 47 ossification centers were recorded. A range and average age for the appearance and fusion of each ossification center were calculated and compared to data available for the rhesus monkey and man.     

Developmental Origins of Pregnancy Loss in the Adult Female Common Marmoset Monkey (Callithrix jacchus)

Background

The impact of the intrauterine environment on the developmental programming of adult female reproductive success is still poorly understood and potentially underestimated. Litter size variation in a nonhuman primate, the common marmoset monkey (Callithrix jacchus), allows us to model the effects of varying intrauterine environments (e.g. nutrient restriction, exposure to male womb-mates) on the risk of losing fetuses in adulthood. Our previous work has characterized the fetuses of triplet pregnancies as experiencing intrauterine nutritional restriction.

Methodology/Principal Findings

We used over a decade of demographic data from the Southwest National Primate Research Center common marmoset colony. We evaluated differences between twin and triplet females in the number of pregnancies they produce and the proportion of those pregnancies that ended in fetal loss. We found that triplet females produced the same number of total offspring as twin females, but lost offspring during pregnancy at a significantly higher rate than did twins (38% vs. 13%, p = 0.02). Regardless of their own birth weight or the sex ratio of the litter the experienced as fetuses, triplet females lost more fetuses than did twins. Females with a male littermate experienced a significant increase in the proportion of stillbirths.

Conclusions/Significance

These striking findings anchor pregnancy loss in the mother’s own fetal environment and development, underscoring a "Womb to Womb" view of the lifecourse and the intergenerational consequences of development. This has important translational implications for understanding the large proportion of human stillbirths that are unexplained. Our findings provide strong evidence that a full understanding of mammalian life history and reproductive biology requires a developmental foundation.     

Birth of Healthy Offspring following ICSI in In Vitro-Matured Common Marmoset (Callithrix jacchus) Oocytes

Intracytoplasmic sperm injection (ICSI), an important method used to treat male subfertility, is applied in the transgenic technology of sperm-mediated gene transfer. However, no study has described successful generation of offspring using ICSI in the common marmoset, a small non-human primate used as a model for biomedical translational research. In this study, we investigated blastocyst development and the subsequent live offspring stages of marmoset oocytes matured in vitro and fertilized by ICSI. To investigate the optimal timing of performing ICSI, corrected immature oocytes were matured in vitro and ICSI was performed at various time points (1–2 h, 2–4 h, 4–6 h, 6–8 h, and 8–10 h after extrusion of the first polar body (PB)). Matured oocytes were then divided randomly into two groups: one was used for in vitro fertilization (IVF) and the other for ICSI. To investigate in vivo development of embryos followed by ICSI, 6-cell- to 8-cell-stage embryos and blastocysts were nonsurgically transferred into recipient marmosets. Although no significant differences were observed in the fertilization rate of blastocysts among ICSI timing after the first PB extrusion, the blastocyst rate at 1–2 h was lowest among groups at 2–4 h, 4–6 h, 6–8 h, and 8–10 h. Comparing ICSI to IVF, the fertilization rates obtained in ICSI were higher than in IVF (p>0.05). No significant difference was noted in the cleaved blastocyst rate between ICSI and IVF. Following the transfer of 37 ICSI blastocysts, 4 of 20 recipients became pregnant, while with the transfer of 21 6-cell- to 8-cell-stage ICSI embryos, 3 of 8 recipients became pregnant. Four healthy offspring were produced and grew normally. These are the first marmoset offspring produced by ICSI, making it an effective fertilization method for marmosets.     

Digestive Efficiency Mediated by Serum Calcium Predicts Bone Mineral Density in the Common Marmoset (Callithrix jacchus)

Two health problems have plagued captive common marmoset (Callithrix jacchus) colonies for nearly as long as those colonies have existed: marmoset wasting syndrome and metabolic bone disease. While marmoset wasting syndrome is explicitly linked to nutrient malabsorption, we propose metabolic bone disease is also linked to nutrient malabsorption, although indirectly. If animals experience negative nutrient balance chronically, critical nutrients may be taken from mineral stores such as the skeleton, thus leaving those stores depleted. We indirectly tested this prediction through an initial investigation of digestive efficiency, as measured by apparent energy digestibility, and serum parameters known to play a part in metabolic bone mineral density of captive common marmoset monkeys. In our initial study on 12 clinically healthy animals, we found a wide range of digestive efficiencies, and subjects with lower digestive efficiency had lower serum vitamin D despite having higher food intakes. A second experiment on 23 subjects including several with suspected bone disease was undertaken to measure digestive and serum parameters, with the addition of a measure of bone mineral density by dual‐energy X‐ray absorptiometry (DEXA). Bone mineral density was positively associated with apparent digestibility of energy, vitamin D, and serum calcium. Further, digestive efficiency was found to predict bone mineral density when mediated by serum calcium. These data indicate that a poor ability to digest and absorb nutrients leads to calcium and vitamin D insufficiency. Vitamin D absorption may be particularly critical for indoor‐housed animals, as opposed to animals in a more natural setting, because vitamin D that would otherwise be synthesized via exposure to sunlight must be absorbed from their diet. If malabsorption persists, metabolic bone disease is a possible consequence in common marmosets. These findings support our hypothesis that both wasting syndrome and metabolic bone disease in captive common marmosets are consequences of inefficient nutrient absorption. Am. J. Primatol. 75:153‐160, 2013. © 2012 Wiley Periodicals, Inc.     

The Response to Novel Foods in Common Marmoset (Callithrix jacchus): The Effects of Different Social Contexts

The main aim of this study was to verify if the response to novel foods in common marmoset (Callithrix jacchus) is influenced by different social settings. This question was tested in three tests involving four social groups. The data regarding the response to novel foods were collected from a total of eight food-naive young. In particular, we explored if direct contacts between food-naive and experienced individuals would have a different effect on the response to novel food by the first, in comparison with a situation in which only visual contacts were allowed. These two different social settings were then compared to a situation in which the response to novel food was not mediated by the experience of group-mates. The results showed a significantly higher number of intervals in which naive young, in social settings, were observed feeding compared with a control test. However, this result was confirmed only in the case of novel foods, not in the case of familiar foods. Furthermore, in the tests in which direct social contacts were allowed, naive young obtained food from group-mates significantly more often in the case of novel than familiar foods. This study suggests that although naive young, when left in pairs, responded quickly to novel foods without the need to observe or interact with experienced group-mates, a process of social facilitation was evident. Such a process was not observed in the case of familiar foods, confirming the hypothesis that social facilitation operates in order to acquire novel foods in the diet of the group in the long term.     

Annual Variation in an Ultradian Component in the Locomotor Activity Rhythm of the Common Marmoset (Callithrix jacchus)

Daily locomotor activity in common marmosets shows a decrease near the middle of the active phase, as observed for captive and wild living animals. An important 8 h period component detected in spectral analysis shows annual variation. To determine the environmental factors that are correlated with this finding, three animals were observed during infant and juvenile stages in outdoor cages under natural temperature, humidity and light cycles. Data were collected from sunrise to sunset, three consecutive days a week, at 5 min intervals randomly distributed inside the clock hours, and were recorded as the total of arbitrary cage sections traveled by the animal each interval. From the Fourier analysis of weekly data series we calculated the relative power of the 8 h period component. Pearson's correlation test showed that the relative power correlates better with environmental temperature and photophase duration than with relative humidity or rainfall. Amplitude but not acrophase showed significant differences (Kruskall-Wallis test, p &lt;0.01) between seasons. These findings suggest that the annual variation in the 8 h period component could result from a temperature modulation on the locomotor activity rhythm.     

Genetic Analysis of Group Composition and Breeding System in a Wild Common Marmoset (Callithrix jacchus) Population

We established pedigree relations in three wild common marmoset social groups for which observational data were available, together with genotypes of some individuals from neighboring groups. Relatedness of 40 individuals were based on 11 microsatellite loci amplified from nDNA obtained noninvasively from plucked hair. The wild marmosets were only half as variable as a captive population characterized previously: 2–6 alleles/locus; H_O = 0.41 and H_E = 0.35. Parentage exclusion probabilities were 61.8% for an offspring and one alleged parent and 90.7% for an offspring with one confirmed and one alleged parent. Each group (n = 5–14 individuals) had two breeding females and 2 adult males. Within each group the infants and reproductively inactive adults were closely related to at least the breeding females; the latter were related to each other as closely as mother/infant pairs or sisters. Relatedness of adult males was lower, indicating recent intergroup dispersal. Genetic data confirm Callithrix jacchus live in relatively stable extended family groups of closely related individuals. Matings occurred preferentially among the least related adults and most infants were fathered by the dominant male. The genetic data are consistent with polygynmonandry as are the field observations. Callithrix have variable mating systems, ranging from monogamy to polyandry to polygyny within social groups plus extragroup copulations; our data provide no evidence for polyandry and are inconclusive with respect to extragroup paternity. Nevertheless, noninvasive multilocus genotyping methods will resolve these questions when longer-term studies of entire populations are undertaken.     

Correction to: Evidence for Dialects in Three Captive Populations of Common Marmosets (Callithrix jacchus)

International Journal of Primatology - A Correction to this paper has been published: https://doi.org/10.1007/s10764-020-00196-x     

Locomotor Activity Rhythm in Free-ranging Common Marmosets (Callithrix jacchus)

Locomotor activity rhythm in the common marmoset has been well described and characterized in captivity, but there is no information about rhythmic aspects of this behavior in free-ranging conditions. To investigate if the locomotor activity rhythm in free-ranging common marmosets is similar to that found in captive common marmosets, we observed reproductive animals from two groups that inhabited at the Experimental Forestry Station, in northeastern Brazil (6°5'S, 35°12'W). From December 1992 to October 1993 we recorded the frequency of locomotor activity by focal animal sampling. The records of locomotor activity were submitted to spectral analysis and to the Cosinor test. Common marmosets showed an average duration of the active phase of 11:37 h ± 13.8 min. Similar to recorded in captive common marmosets, circadian rhythmicity was found with peaks in the morning, and in the afternoon, characterizing a bimodal pattern with peaks separated by 8 hours. The presence of a rhythmic pattern in the locomotor activity in free-ranging and captive common marmosets suggests that this pattern is not determined by the environmental conditions in captivity, but it has a endogenous component that play an important role for adaptation of this species to different conditions.     

Severe Intragroup Aggressions in Captive Common Marmosets (Callithrix jacchus)

Members of captive colonies of common marmosets (Callithrix jacchus), a small New World primate, can occasionally be victims of repeated, and potentially fatal, attacks by a family-mate. This study examined the records of a colony, looking for past instances of such aggressions. The aim was to better understand the possible causes underlying this phenomenon and to identify variables that could minimize their occurrence. The results showed that both males and females behaved as aggressors at the same rate, but females attacked just females, whereas males attacked both males and females. Most aggressions involved siblings and occurred at a higher rate when the cages were occupied by more than 5 nonhuman animals. The influence of group size was significant only when females behaved as aggressors. This study suggests that several factors can determine the occurrence of these aggressions and that their relative importance depends on the sex of the aggressor. The results of this study highlight the importance of collecting further data to verify whether crowding in a confined space, rather than group size per se, affects the incidence of these aggressions.     

Treatment of Giardiasis in Common Marmosets (Callithrix jacchus) with Tinidazole

Giardia intestinalis is a common protozoan parasite that can infect many laboratory animal primates, although its role as a contributor to the induction of gastrointestinal disease remains unclear. This study sought to investigate the prevalence of Giardia in a colony of common marmosets by using a Giardia antigen-capture assay and to address the possible eradication of this infection by using tinidazole, an antiprotozoal similar to metronidazole but requiring fewer doses. Among 31 colony marmosets, 13 (42%) were positive for Giardia. Two doses of oral tinidazole eliminated the infection in all animals. Repeat testing of the 13 Giardia-positive monkeys 1 y later showed that 11 remained negative and that treated animals had a significant increase in weight at 1 y. Giardia antigen is common in common marmoset feces, and treatment using oral tinidazole is possible and highly effective.Giardia intestinalis is a common zoonotic protozoan parasite causing diarrhea in humans and animals worldwide. Infection usually results from contact with the feces of an infected host or drinking water contaminated with Giardia cysts. As few as 10 cysts are necessary for infection in human subjects.³⁶ Giardia causes both an acute disease and a chronic asymptomatic state. The most common clinical signs of acute disease are diarrhea, flatulence, foul stool, and abdominal cramps. In addition, Giardia has been implicated as a cause of cognitive impairment and stunted growth in infected children in developing countries.^1,2,29Giardiasis has previously been reported to occur in a variety of laboratory primates, including several species of neotropical monkeys such as marmosets and squirrel monkeys.^13,20,28 Such infections may pose a zoonotic risk to animal handlers and potentially affect colony health. Common marmosets (Callithrix jacchus) frequently manifest chronic wasting and inflammatory bowel disease known as ‘wasting marmoset syndrome.’ The etiology of the intestinal disease is unknown, but marmosets often present clinically with skeletal muscle atrophy, marked weight loss, alopecia, and a history of intermittent diarrhea.^{5,11,16,21,35} Likely no single infectious agent or nutritional deficiency causes the clinical spectrum, but rather a combination of factors result in antigenic stimulation of the intestinal tract resulting in the chronic disease. The presence of Giardia cannot be ruled out as a cofactor.Several studies have compared various diagnostic tools for detection of Giardia in fecal specimens. Multiple fecal tests for Giardial colonization are available, including antigen-detection enzyme immunoassays, immunochromatographic strips, and microscopy of wet-mounted stool after fecal flotation.^9,24,37,38 Enzyme immunoassays are a rapid and precise tool for detecting Giardia in fecal specimens; test sensitivities and specificities have approached 100% in several studies.^9,22,24,37 In addition, these studies have indicated that repeat stool sampling on different days may increase the yield of testing, because organisms are variably shed.^9,14,36 The specific recommendation is to test 3 samples on alternate days or 3 samples within a 10-d span.³⁸Treatment options for Giardia infection are varied and include metronidazole, albendazole, quinacrine, furazolidone, and several other nitroimidazoles including tinidazole, secnidazole, ornidazole, and nimorazole.¹⁵ The most common treatment choice in veterinary medicine is metronidazole; however, this drug requires 5 to 8 d of treatment and ensuring animal compliance is difficult.²⁷ Several of metronidazole''s structural analogues, including tinidazole, are used as a single dose in the treatment of Giardiasis in humans with high cure rates (approximately 90%) and low complications.^3,8,10,26,39In this study we examined the use of a commercially available antigen-capture assay to diagnose Giardiasis in a large breeding colony of common marmosets. To address possible eradication of the infection, we describe the safe use and efficacy of tinidazole as a new treatment option in this species.     

Body Weight of Wild and Captive Common Marmosets (Callithrix jacchus)

Captive studies and occasional trappings of wild individuals indicate that callitrichids have small size and body weight and lack sexual dimorphism. We compared body weights of captive and wild Callithrix jacchus obtained by repeatedly weighing subjects from two populations in Brazil. We obtained captive data by routinely weighing 138 individuals from the Universidade Federal do Rio Grande do Norte colony and wild data via regular trapping of 243 individuals in 15 free-ranging groups from IBAMA's field site in Nísia Floresta. We assigned all subjects to one of four age classes—infant, juvenile, subadult, and adult—according to their birth dates or size, reproductive status, and dental development. There is no significant difference between males and females in any of the four age classes, but captive subjects were heavier than wild ones in all age classes but infant. Reproductive and nonreproductive adult females showed no statistical difference in weight. These results accord with previous reports of lack of body size sexual dimorphism in common marmosets and suggest that differences between wild and captive common marmosets are not constitutional, but are instead a consequence of diet and physical activity. The absence of weight difference between reproductive and nonreproductive females suggests that any possible advantage from high rank is outweighed by the costs of reproduction in common marmosets.     

Circadian Rhythmicity of Grooming Behavior During Pregnancy in a Marmoset (Callithrix jacchus) Family Group in Captivity

One family group of common marmoset (Callithrix jacchus) was observed in captivity under natural environmental conditions to describe the circadian pattern of grooming behavior during the pregnancy of the reproductive female. The animals were observed by focal sampling, from the beginning to the end of animal's active phase, during two non-consecutive days per week, from August to November 1996. The duration of self-grooming and social grooming behavior of each animal were totalled each hour. The time series obtained were analyzed by Cluster Analysis and by Sin gle Cosinor Analysis. The daily patterns of grooming behavior during pregnancy were divided by Cluster Analysis into two stages, one corresponding to the second half of the third month of pregnancy, and the other from the last half of the third month to the last week before parturition. The confidence intervals of acrophases overlapped in all stages. The acrophases values of the first stage occurred between 10:09 h and 11:00 h, and on the second stage between 8:51 h and 11:13 h. The daily pattern of grooming behavior of adult male and female showed a trend to increase in the first hours of the active phase. This trend to advance grooming behavior during the second stage of pregnancy may be due to an effect of the coming parturition itself or to seasonal influences.