Mother-infant interactions of wild-born,individually-caged cynomolgus monkeys (Macaca fascicularis) during the first 14 weeks of infant life

This study documents age-related changes in the interactions of wild-born cynomolgus macaque mothers and their infants living in individual cages during the first 14 weeks of infant life. Body contact between mother and infant, maternal holding, and infant sucking were found to decrease, and the mothers showed an increased frequency of aggression toward their infants with age. These results were broadly similar to those reported for mother-infant interactions in other macaques living in social groups. Nevertheless, a clear difference between the present cynomolgus macaques and other macaques in social groups was apparent. The cynomolgus macaque mothers tended to permit their infants to move about freely without displaying maternal protectiveness such as restraint or retrieval, unlike other macaque mothers in social groups. Such maternal behaviors might derive from the experience of living in individual cages for many years and the relative safety of living in individual cages. The lack of maternal restraint and retrieval could be responsible for the observed sex differences in behavior: male infants moved more actively, and broke, and made contact with their mothers more frequently than did female infants. Moreover, mothers of female infants held and groomed them more frequently and were less aggressive toward them.     

Perception of stimuli presented as photographic slides in cynomolgus macaques (Macaca fascicularis)

The present study was designed to assess a monkey's perception of specific visual stimuli by measuring both the behavioral responses and duration of attention to the presentation of photographic slides. Five adult male cynomolgus macaques (Macaca fascicularis) were placed individually in an open field apparatus and presented a series of slides consisting of apples, a gorilla mask, a collage of colors, a human being, and a plain field. The slide of the gorilla mask followed by that of the human being received the most attention while the plain field received the least. In addition, the gorilla mask and human being elicited a range of behavioral responses with the higher ranking animals displaying a greater number of aggressive responses and the lower ranking animals displaying a greater number of submissive gestures. Taken together, these data would suggest that the slides of the gorilla mask and the human being were perceived by the monkeys as threatening. These results are consistent with a continuing theme observed among a number of studies of primate social perception — namely, that potentially threatening stimuli are a significant determinant of visual observing.     

Isolation and culture of hepatocytes from the cynomolgus monkey (Macaca fascicularis)

Summary Isolation and culture techniques for hepatocytes from whole livers of the cynomolgus monkey,Macaca fascicularis, are described. Hepatocytes were isolated by two-step perfusion of livers, using collagenase with hyaluronidase; fructose and trypsin inhibitor were included to reduce cell loss. Yields from a single liver average 4×10⁹ cells with viabilities of 90.8±5.7%. Cells, plated on collagen substrates, were assessed for changes in morphology and various marker enzyme activities over a period of 7 d in culture. Cells exhibited a morphology similar to that observed for this species in vivo; little change in attached and spread cells was observed over the length of time monitored. Enzyme activities for catalase, succinate dehydrogenase, and tyrosine aminotransferase were observed to decrease significantly (though considerable activity remained), whereas acid phosphatase and 5′-nucleotide phosphodiesterase remained unchange. Activity of cytochrome P-450 reductase was observed to increase slightly for the first 2 d, then decrease to about 60% of initial levels. Activity of α-mannosidase was stable for 4 d but was observed to be increased at Day 7. Cells were observed to retain metabolic responsiveness demonstrated by glucose production by both gluconeogenesis and glycogenolysis in response to glucagon stimulation. The monkey hepatocytes obtained by methods described here thus retain hepatocellular morphology and activity through at least 1 wk in culture without medium or culture modification.     

The innervation of the myometrium of the cynomolgus monkey (Macaca fascicularis)

Summary The autonomic innervation of the myometrium of Macaca fascicularis consists of bundles of unmyelinated nerve fibres running between the smooth muscle cells, and is therefore considered to be of the fascicular (= unitary) type. Close contacts between nerve fibres and smooth muscle fibres were not found. Modification of the chromaffin method according to Tranzer and Richards made it possible to visualize the heterogeneity of the nerve fibres in a single bundle. The following fibre types were found to coexist: (1) noradrenergic fibres containing synaptic vesicles with a dense granule, (2) cholinergic fibres containing empty synaptic vesicles, and (3) non-adrenergic noncholinergic (NANC) fibres containing only or predominantly large dense-cored vesicles, which do not react with this method. Noradrenergic fibres are the most numerous (around 60%), followed by NANC fibres (30%) and cholinergic elements (around 10%). The distribution of these three types is similar in the cervix, the isthmus and the body of the uterus in pregnant and non-pregnant females.     

Ovum recovery and in vitro fertilization in crab-eating monkeys (Macaca fascicularis)

A total of 301 oocytes were recovered from crab-eating monkeys and subjected to insemination in vitro resulting in two fertilized ova. Sixteen monkeys in 24 cycles received 37.5 IU of hMG daily from the second day of the menstrual cycle for 7 to 10 days. Oocytes were recovered under laparotomy at 20 to 49 hr after administration of 1,000–1,500 IU of hCG. The maturation rate of the recovered oocytes was 24.2% as judged from morphological criteria under the light microscope. With additional maturation culture, the rate increased to 36.2%. The matured oocytes were inseminated at 3 to 4 hr after aspiration using homologous spermatozoa which had been capacitated in vitro. Two oocytes were judged as being fertilized based on the presence of 3 and 5 pronuclei, respectively, when examined 12 hr after the insemination. This is the first report of in vitro fertilized ova in nonhuman primates in Japan.     

Development of the brain in staged embryos of the long-tailed monkey (Macaca fascicularis)

The external characteristics and successive morphological changes of the brain and its derivatives were studied in 69 long-tailed monkey embryos representing developmental stages 8 through 16. This morphogenesis follows a similar pattern to those of the rhesus, baboon, and human. Minor differences in the temporal sequence of specific developmental events include: 1) otic disc, adenohypophyseal pouch, and hippocampal internal sulcus formation in the long-tailed macaque occur at stage 10, stage 11, and after stage 16 respectively, which is comparable to human stages 9, 10, and 16; 2) formation of the trigeminal primordium and the motor root of the trigeminal nerve and evagination of the neurohypophysis occur at stage 12, stage 14, and stage 15, while in the human embryo these features are observed at stage 14, stage 15, and stage 16, respectively; and 3) closure of the lens pore, like in the rhesus monkey, occurs during stage 15, while in the baboon and human it takes place during stage 14. These temporal differences in the embryonic period are important factors to be taken into consideration in any embryological and teratological studies when usingM. fascicularis as a primate model.     

Mouse/human chimeric anti-HIV-1 gp120 antibody to the principal neutralizing determinant: Tolerability and pharmacokinetics in cynomolgus monkeys,Macaca fascicularis

In preparing for testing a pharmaceutical grade preparation of chimeric (mouse/human) antibody CGP 47 439 in HIV-1 infected individuals, it was administered toMacaca fascicularis (cynomolgus) monkeys to study tolerability, immunogenicity and pharmacokinetics. Four groups of monkeys, three males and three females per group, received respectively four infusions of 0, 1.43, 4.3, and 14.3 mg of CGP 47 4391 kg body weight at one-week intervals. The chimeric antibody induced no fever, was tolerated well throughout the 50-day observation period, elicited no tissue damage and no anti-antibody response. The pharmacokinetic profile was similar at all dose levels with a mean T_1/2 of 14.2 h (range 11.8–19.3 h) and a mean T_1/2 of 172.6h (range 137.2–220.5h). Following four successive antibody infusions serum concentrations of CGP 47 439 increased without reaching a steady state, and its measured concentrations were comparable to the simulated values. Collectively the study has provided safety and pharmacokinetic data that would allow human studies with this antibody in AIDS patients.     

Feeding ecology of the long-tailed macaque(Macaca fascicularis) in Kalimantan Tengah,Indonesia

I studied long- tailed macaque (Macaca fascicularis)feeding behavior and ecology as part of a larger behavioral ecological study at the Natai Lengkuas Station, Tanjung Puting National Park, Kalimantan Tengah, Indonesia. I collected data on feeding behavior via scan sampling of all visible individuals in the focal group (approximately 800 observation hours). I established vegetational plots and monitored them monthly to determine food availability and abundance. I found long- tailed macaques to be primarily frugivorous;leaves, flowers, insects and bark provided the remainder of the diet. They used at least 33 plant species as food sources, but >60% of the diet was provided by only 5 species. Based on previous vegetational analyses, these tree species were among the highest in relative density. However, selection ratios for 19 food species indicate that 13 of them were selected more often than expected. Long- tailed macaques appear to be selective feeders but can exploit a variety of food sources during periods of food scarcity.     

Alternative seed-handling strategies in primates: seed-spitting by long-tailed macaques (Macaca fascicularis)

Summary The seeds in fruits consumed by primates may be chewed and digested, swallowed and defecated intact, or separated from the flesh and spat out. We show by a combination of close field observations and experiments with caged animals, that long-tailed macaques (Macaca fascicularis) have a remarkably low threshold of 3–4 mm for swallowing seeds and also that wild macaques rarely break them. The seeds of 69% of the ripe fruit species eaten are spat out intact or cleaned outside the mouth and dropped. Seed-spitting significantly reduces the swallowed food bulk and may lessen the risk of releasing seed toxins during mastication. However, it requires that even small fruits are processed in the mouth one or a few at a time. We suggest that fruit storage in the cheek pouches of cercopithecine monkeys allows them to spit seeds individually without excessively slowing fruit intake while feeding on patchily distributed fruit. In contrast, Apes and New World monkeys apparently swallow and defecate most ripe seeds in their diet and colobine monkeys break and digest them, detoxifying seed defenses by bacterial fermentation.     

Genetic characterization of long-tailed macaques (Macaca fascicularis) on Tabuan Island Indonesia

Protein and mitochondrial DNA variations (D-loop region PCR-RFLP) were analyzed for 7 serum and 40 clot samples collected from long-tailed macaques (Macaca fascicularis) living on Tabuan Island, Indonesia. Protein polymorphisms were examined electrophoretically for 5 and 12 kinds of protein in serum and erythrocytes, respectively. Each of the protein loci tested showed a monomorphic pattern. Polymorphisms were detected in the analysis of the D-loop-containing region of mtDNA (PCR-RFLP) using 32 restriction endonucleases. Two haplotypes, differing 1.03% in sequence divergence were observed, and both were previously undetected in other local populations. Based on genetic features and differences in pelage color as outlined inFooden's (1995) morphological analysis, the present results suggest that long-tailed macaques on Tabuan Island are a unique population. From the genetic analyses performed here, Tabuan monkeys are considered to be the same species group as those populations of Sumatra and Java (Fooden, 1995).     

The extensor and peroneal muscles of the crab-eating monkey (Macaca fascicularis)

On the basis of 60 hindlimbs of 30 crab-eating monkeys (Macaca fascicularis) of each sex, the morphology of the crural extensor and the peroneal group of leg muscles is described and some functional indices are calculated. For the attachments and measurements of the muscles, the results obtained in this study generally agreed with those of otherMacaca species. The crural extensor has minor differences. Some anomalous modes of insertion are observed in the peroneal group. These results indicate the phyletically labile state of the peronei in the crab-eating monkey.     

New Type of Puzzle-Task Finger Maze Learning in Macaca fascicularis

In order to easily estimate the global cognitive ability of nonhuman primates, we developed a 4-step noncorrection-method-type finger maze (4FM) based on the standard puzzle feeder. We tested 7 experimentally naïve long-tailed macaques (Macaca fascicularis) to assess the validity of the apparatus and the testing procedure. The most notable difference between the 4FM and the standard puzzle feeder is the presence of an error box. There is a hole at both ends of each step. One hole of each step is connected to the lower step or feeding box. The other hole of each step is connected to an error box. The monkey had to move the reward into the feeding box without dropping it into the error box and to retrieve the reward from the feeding box. Task difficulties could be controlled by deciding on which step to place the food reward at the beginning of the trial. All the monkeys could complete the tasks without food/water deprivation and pretraining. The results suggest that the 4FM is a suitable device to assess the cognitive ability of the monkeys simply, easily, and objectively.     

Responses to a snake model in captive crab-eating macaques (Macaca fascicularis) and captive tufted capuchins (Cebus apella)

The responses to a model snake in captive crab-eating macaques (Macaca fascicularis) and captive tufted capuchins (Cebus apella) were investigated. In both species the amount of fear behavior was higher in the presence of the model than during baseline conditions. Unlike the macaques, in the capuchins the frequency of these behaviors decreased across trials. In the two species the amount of explorative and manipulative behaviors and the use of space were also different. Unlike macaques, all capuchins manipulated the objects available in the testing room, and three subjects contacted the apparatus by using objects. Macaques did not show significant preferences for any particular part of the testing room. In both conditions, capuchins used the floor more than macaques did. Further, capuchins increased the use of the floor across experimental trials. Latency to reach the floor was higher in macaques than in capuchins. When on the floor, capuchins spent most of the time close to the snake apparatus. It is proposed that the behavioral differences between capuchins and macaques in the responses to a potential predator indicate that capuchins have a greater propensity to explore and to contact the novel stimulus directly, or by means of objects, than macaques do. These tendencies may lead to the exploitation of novel features in the environment.     

Enzootic hepatitis A infection in cynomolgus monkeys (Macaca fascicularis)

During a toxicology study in cynomolgus (long-tailed or crab-eating) monkeys (Macaca fascicularis), a randomly distributed incidence of significantly increased hepatic enzyme activity was observed. Premedication hepatic enzyme activity in all monkeys of this study was normal, but increased alanine aminotransferase (ALT) activity was found in 4 of the 24 animals 2 weeks after initiation of the study and in 10 of 24 at 4 weeks. A drug-related effect was considered unlikely initially because the increases were not doserelated, and a 3-year review of 655 cynomolgus monkeys revealed a 15–20% incidence of increased hepatic enzyme activity. Good correlation was subsequently established between increased hepatic enzyme activity, active hepatitis A virus (HAV) infection, and histomorphologic confirmation of hepatitis (chronic periportal inflammation). Follow-up viral serodiagnostic screening of resident macaques revealed an overall incidence of anti-HAV IgG in 80% (155/193) of cynomolgus and in 70% (14/20) of rhesus monkeys. Serial screening demonstrated that several initially negative monkeys became seropositive for anti-HAV IgG, and a few acquired active infection (anti-HAV IgM). Among newly acquired cynomolgus monkeys, 2.5% (2/80) had an acute HAV infection, and 35% (28/80) eventually tested positive for anti-HAV IgG while quarantined in the primate facility. The characterization of an enzootic HAV infection in incoming monkeys posed a significant risk for the primate colony and handlers. Rigorous sanitation, isolation, and quarantine procedures, including personnel training and additional protective clothing for personnel working in the primate colony, reduced tho potential for transmission and arrested the outbreak. Experimenters should be cautious in ascribing toxicity to a test article based solely on increased hepatic enzyme activity associated with chronic periportal inflammation.     

Adoption success under single-cage conditions by cynomolgus macaque mothers (Macaca fascicularis)

Retrospective data were studied to determine the success of experimental adoption by 122 cynomolgus macaques kept in individual cages at the Tsukuba Primate Center. The four types of adoption procedures included (1) singletons adopted after weaning (SAW)—21 cases; (2) singletons adopted after stillbirth (SAS)—20 cases; (3) singletons adopted after forced separation of the biological infant during the nursing period (SAF)—4 cases; and (4) adoption of a second “twin” while still nursing a biological infant (TA)—77 cases. Adoption was defined as successful if the mother nursed the foster infants for at least 7 days. Success rates (percent) were 47.6 for SAW, 75 for SAS, 100 for SAF, and 74 for TA. Success was affected by the postbirth stage of foster mothers at adoption but not by foster infant age or sex.     

Social Behavior of Captive Indochinese and Insular Long-Tailed Macaques (Macaca fascicularis) Following Transfer to a New Facility

There are measurable differences in behavior, physiology, social organization, and geographic distribution within and between various species of macaques. We collected information on the social behavior of captive Macaca fascicularis from Indochina and island populations after they had been transferred to a new environment and new social groups. While some changes in behavior occurred over time, we found no decrease in high levels of agonistic behaviors. We interpret this finding in light of previous research reporting that long-tailed macaques show low levels of habitatuation to novelty and are highly aroused in comparison to other macaque species. We found predictable differences in behavior for males, females, and infants, in which infants played more, females had higher levels of contact proximity to other individuals, and males exhibited more sexual and threat behavior. A comparison of social behavior in long-tailed macaques from different origins indicate that Indochinese macaques are generally less affiliative and Indochinese males are more aggressive than their insular counterparts. Differences among macaque species, and within the fascicularis-group, should be considered in management of captive colonies and when interpreting research data.     

Developmental trends in mother-infant interactions in Java macaques (Macaca fascicularis) during the first ten days of life

In this study on Java macaques themain characteristics of the mother-infant relationship during infants' first 10 days have been elaborated. Early mother and infant behaviour is described and early interactions between mothers and their young have been analyzed. Observations and trend analyses of early behaviours like ventro-ventral contact behaviour, being on nipple, and explorative behaviour, make it clear that Java macaque infants play an important role in the early development of mother-infant interactions. In contrast to the rhesus and the stumptailed infant, the young Java macaque appears to develop a great variety of behaviours in its first 10 days. Whereas the infant's behaviour gains in complexity and social initiative, the mother becomes increasingly more passive and self-oriented. Early mother-infant interaction can be characterized by mutual adjustment.     

Inhibin-like and gonadotropin-like immunoreactivity in pituitary cells of male monkeys (Macaca fascicularis,Macaca mulatta)

Summary Inhibin-like immunoreactivity was detected by immunocytochemistry in the pituitaries of untreated male crab-eating macaques (cynomolgus monkey) and rhesus monkeys, in rhesus monkeys actively immunized against FSH, and in one orchidectomized crab-cating macaque. Localizations were performed by the immunogold-silver staining with 5-nm colloidal gold-conjugated second or third antibodies and by the alkaline phosphatase-anti-alkaline-phosphatase technique. Two different inhibin-specific antisera, raised against the -subunit or the entire inhibin molecule, provided identical staining patterns. Positive label was confined to the pars distalis of the pituitary and occurred exclusively in the cytoplasm of morphologically different cell types throughout the pars distalis in all pituitaries. Staining was most prominent in clusters of chromophobic cells. The presence of inhibin-like activity in the pituitary of an orchidectomized monkey with undetectable serum inhibin levels suggests that inhibin is produced within the pituitary gland. Co-localization studies for the -subunits of the gonadotropic hormones revealed that on average 82% of the gonadotropes were bihormonal. Using the same protocol, co-localization of inhibin-like activity with gonadotropin-like immunoreactivity revealed only a small degree of common distribution (<15%). Inhibinpositive cells were frequently in close proximity to gonadotropic cells and, thus, paracrine effects of inhibin on gonadotropin-synthesizing cells are conceivable.     

Breeding seasonality among long-tailed macaques,Macaca fascicularis,in Peninsular Malaysia

The females among more than 7000 long-tailed macaques, Macaca fascicularis,that had recently been trapped for biomedical research were examined for pregnancy. The animals were taken from the wild in the north of Peninsular Malaysia during the period 1978 to 1979 and in the south from 1978 to 1982. Both samples showed some pregnancies in all months of the year and a peak during the months of May to July. Additional peaks in other months in some years showed that, while the midyear pregnancy peak was significant, it was not as marked a phenomenon as is the breeding peak in many other species. This might be due to the complexity and unpredictability of both the climate and the macaques’ food supply in Peninsular Malaysia. The additional peaks also illustrated the pitfalls of generalizing about breeding seasonality on the basis of small samples.