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The effects of cage size and complexity on the behaviour of captive common marmosets, Callithrix jacchus jacchus.

Conditions of captivity of primates used in biomedical research may have deleterious effects on the welfare of the animals and consequently on the reliability of the research. We investigated the effects of cage size and cage complexity, two fundamental characteristics of captive conditions, on the behaviour of the common marmoset (Callithrix jacchus jacchus). We found an increase in the general level of activity and significant variation in the frequencies of specific behaviours with an increase in cage size and also with cage complexity. Stereotyped behaviours, which occurred in the small cages, were never exhibited in the large cages. The effect of the novelty of the changed conditions was also assessed and found to be significant for some behaviours. We also measured the time taken to capture an animal, a task frequently performed by the animal technician, under the various cage conditions. Capture time increased significantly in the larger cages, but the overall effect of the changes to the marmosets' housing conditions on the animal technician's work was not regarded as substantial. We conclude that the welfare of captive marmosets is enhanced by the provision of larger and more complex cages, and that such cages do not significantly affect the efficiency of the research laboratory.

Animal Welfare↗

Anaesthesia of the common marmoset (Callithrix jacchus) using continuous intravenous infusion of alphaxalone/alphadalone.

A safe means of anaesthetizing common marmosets (Callithrix jacchus) for a study using magnetic resonance imaging (MRI) to investigate cerebral ischaemia was required. Continuous infusion of alphaxalone/alphadalone was used to anaesthetize 37 marmosets for non-recovery and recovery experiments. This was found to give safe, reliable anaesthesia when coupled with pulse oximetry and electrocardiographic (ECG) monitoring.

Alfaxalone Alfadolone Mixture↗

Spontaneous lesions detected in the common cotton-eared marmosets (Callithrix jacchus).

Spontaneous lesions in the common cotton-eared marmosets (Callithrix jacchus), which were maintained in our laboratory or were purchased from a commercial breeder, were examined histopathologically. There were 39 males and 22 females, between the ages of 7 and 100 months. The most interesting finding noted was extramedullary hematopoiesis, which was observed in the mesenteric lymph nodes, spleen, liver, kidney, adrenal gland, and cerebral choroid plexus. Megakaryocytes were frequently observed in the blood vessels of the alveolar wall of the lung. It is important to distinguish extramedullary hematopoiesis which occurs accidentally from that induced by repeated blood sampling or that resulting from administration of chemicals. Extramedullary hematopoiesis was easily distinguishable from inflammation or hematopoietic neoplasms, because of the various cellular elements present, including megakaryocytes. In the weak and dead animals, enterocolitis was found at a high frequency. Other common findings included thymic involution, prominent vacuolation of liver sinusoidal cells, which were probably Ito-cells, and basophilic changes in the renal tubular epithelium. The liver and the kidney are frequently affected by toxic effects of chemicals. Therefore, it is important to distinguish the toxic changes from the spontaneous ones. Proliferation of apocrine glands in the cervical skin was an interesting finding; however, neither inflammatory change nor cellular atypia was noted in this structure. Although its function is unclear, this structure may be a marmoset-specific tissue.

Adrenal Glands↗

Effects of click intensity and frequency on the brain-stem auditory evoked potentials in the common marmoset (Callithrix jacchus).

Brain-stem auditory evoked potentials (BAEPs) were recorded in 20 common marmosets (Callithrix jacchus) to investigate the effects of click frequency up to 99 kHz, in consideration of the higher hearing range of the marmoset, and intensity on wave forms and peak latencies. According to the results of BAEP recordings at frequencies of 4, 32, and 99 kHz, the number of components recorded was affected by the stimulus intensity and the clicks at an intensity of 80 dB peak equivalent sound pressure level (pe SPL) had the maximum number of clear components. Therefore, it was indicated that click stimulations at an intensity of 80 dB pe SPL over a broad range of frequencies appears to be useful for recording the maximum number of components in marmosets and may increase the information obtainable from BAEPs. BAEP latencies were prolonged as the stimulus intensity decreased from 100 to 50 dB pe SPL. The effects of stimulus frequency on the wave latencies and amplitudes in response to 80 dB pe SPL at frequencies between 0.5 and 99 kHz revealed various changes: the amplitude of wave I increased at 16 and 32 kHz, but that of waves III and V increased at 4-8 and 64-99 kHz. These increases in amplitudes of the waves may correlate with higher synchronous activity of the peripheral or central auditory pathways.

Acoustic Stimulation↗

Distribution of protein gene product 9.5-immunopositive and NADPH-diaphorase-positive neurons in the common marmoset (Callithrix jacchus) accessory olfactory bulb.

The principal center of the accessory olfactory system is the accessory olfactory bulb (AOB). In primates, simians are divided into two groups, New and Old World monkeys, and the AOB is present in only New World monkeys. The common marmoset (Callithrix jacchus) is a species of New World monkey. Although the morphology of the common marmoset AOB has been demonstrated, the distribution patterns of the mitral/tufted and granule cells of the AOB remain unclear. In the present study, therefore, the distribution of the mitral/tufted and granule cells in the common marmoset AOB was examined using two histochemical markers including immuno-staining for protein gene product (PGP) 9.5 and NADPH-diaphorase staining. The vomeronasal nerves, gomeruli and mitral/tufted cells showed PGP 9.5-immunoreactivity. The mitral/tufted cells were arranged in only one or two rows along the margin of the glomerular layer to form the mitral/tufted cell layer (MTL). Since the mitral/tufted cells occurred sparsely in the common marmoset, the MTL was illegible. NADPH-diaphorase reactivity was primarily detected in the rostral and caudal areas of the AOB. In these areas, granule cells showed NADPH-diaphorase reactivity. Since the granule cells were sparse, the common marmoset AOB displayed less-developed granule cell layer. Although the functional significance of the AOB remains to be solved in the common marmoset, small-sized and less-laminated AOB may show that sexual behavior of the common marmoset has lesser dependence on the accessory olfactory system.

Animals↗

Fatal Herpes simplex infection in a group of common marmosets (Callithrix jacchus).

An outbreak of classical herpetic infection causing vesicoulcerative stomatitis in a family group (eight animals) of Callithrix jacchus is described. In all eight infected animals, human herpesvirus 1 (HHV-1) was identified as the causative agent. This was confirmed by histologic, immunohistologic, and molecular biologic investigations, as well as by virus isolation. The clinical picture, the macroscopic appearance, and the histologic results indicated a herpes infection as the cause of mortality. Alterations of the oral mucous membranes were erosive to ulcerative with typical intranuclear inclusions. Immunohistologic and molecular biologic techniques clearly identified the HHV-1 virus and excluded other possible primate herpesviruses such as B-virus, SA8, HVP-2, and Herpes tamarinus. The significance of this herpesvirus infection for colony management is discussed.

Animals↗

Anemia, myopathy, and pansteatitis in vitamin E-deficient captive marmosets (Callithrix spp.).

Five young adult pet marmosets (Callithrix spp.) were presented with weight loss (5/5); fecal retention (3/5); diarrhea (2/5); impaired locomotion (3/5); anemia (4/4); hypoproteinemia or hypoalbuminemia (3/4); elevations of creatine phosphokinase, lactic dehydrogenase, and alanine aminotransferase (3/4); and renal failure with hypercholesterolemia (2/4). All anemic marmosets had low serum vitamin E levels. The anemia responded to vitamin E and selenium therapy in two marmosets. One of the five marmosets died before presentation, and two others died despite therapy. The two marmosets necropsied had degenerative myopathy, pyogranulomatous pansteatitis, and increased erythrophagocytosis and hemosiderosis. The striated muscle and adipose tissue of both marmosets were negative for coxsackievirus ribonucleic acid by in situ hybridization. These findings suggest that vitamin E deficiency may be involved in the development of anemia, myopathy, and steatitis in callitrichids; however, in some marmosets, underlying diseases such as chronic colitis may have influenced the development of anemia and impaired vitamin E status.

Adipose Tissue↗

Natural occurrence of amyloid-Abeta deposits in the brain of young common marmosets (Callithrix jacchus): a morphological and immunohistochemical evaluation.

The spontaneous incidence and distribution of beta-amyloid (Abeta) peptide and argyrophilic deposits were extensively investigated in the common marmoset (Callithrix jacchus). In 12 young marmosets, up to 67 coronal step sections were evaluated in the brain. One brain hemisphere was stained by a silver method and also immunohistochemically with a primary antibody directed against Abeta. Argyrophilic deposits were observed in 4 (33%) of 12 animals with silver stain, and Abeta was seen in 4 (33%) of 12 animals with anti-4G8 antibody. The overall incidence was 6 (50%) of 12 affected animals. These results show that the young marmoset brain displays Abeta deposits at a low incidence and that immunohistochemistry is more suitable than silver staining to reveal rare amyloid plaques. These findings also provide evidence for the early onset of amyloid deposits in marmoset brain.

Amyloid beta-Peptides↗

Comparison of androgen receptor and oestrogen receptor beta immunoexpression in the testes of the common marmoset (Callithrix jacchus) from birth to adulthood: low androgen receptor immunoexpression in Sertoli cells during the neonatal increase in testosterone concentrations.

The aims of this study were: (i) to investigate the cellular immunoexpression of androgen receptor and oestrogen receptor beta in the testes of the common marmoset (Callithrix jacchus) during neonatal life compared with their expression at later ages; (ii) to establish whether neonatal marmoset Sertoli cells are targets for androgens or oestrogens or both; and (iii) to investigate the relationship between neonatal plasma testosterone concentrations and androgen receptor immunoexpression by abolishing the neonatal testosterone surge with a potent GnRH antagonist. Androgen receptor and oestrogen receptor beta immunoexpression were evaluated in neonatal animals aged 1-4 days, 4 weeks and 6 weeks, and compared with immunoexpression in animals aged 18-22 weeks (early infancy), 35 weeks (late infancy), 58-62 weeks (late pubertal) and > 100 weeks (adult). Immunoexpression of androgen receptor in the reproductive tract was also evaluated at each age. Sertoli cell immunoexpression of androgen receptor was weak or absent in neonatal animals, but increased substantially in infant animals, reaching adult levels by the end of infancy. In contrast, immunoexpression of androgen receptor during the neonatal period was strong in testicular interstitial cells and very strong in epithelial cell nuclei throughout the reproductive tract, and did not change greatly with age in these cells or tissues. Similarly, immunoexpression of oestrogen receptor beta was prominent in many Sertoli cells and in the germ cells of neonatal animals, and was relatively constant throughout life. Weak immunoexpression of androgen receptor in neonatal Sertoli cells was associated with high plasma testosterone concentrations (2.7-5.5 ng ml(-1)), whereas strong Sertoli cell immunoexpression was associated with baseline (approximately 0.12 ng ml(-1)) testosterone concentrations in infant animals and with > 10 ng ml(-1) in late pubertal and adult animals. Immunoexpression of androgen receptor and oestrogen receptor beta was also evaluated in co-twin males aged 4 and 35 weeks, after treatment from birth to 4 weeks or from week 25 to week 35, respectively, with either vehicle or with GnRH antagonist at a dose known to suppress the neonatal testosterone surge completely. Only GnRH antagonist treatment during weeks 25-35 reduced androgen receptor immunoexpression, whereas immunoexpression of oestrogen receptor beta was unaffected by treatment during either period. On the basis of these findings it is suggested that: (i) neonatal marmoset Sertoli cells may be targets primarily for oestrogens rather than androgens; (ii) androgen receptor expression in the testes of neonatal and infant marmosets is not regulated in a straightforward way by testosterone; and (iii) high neonatal concentrations of plasma testosterone are not absolutely necessary for expression of androgen receptor in marmoset testes at this time.

Aging↗

Anatomical structure and surface epithelial distribution in the nasal cavity of the common cotton-eared marmoset (Callithrix jacchus).

To validate use of the common cotton-eared marmoset (Callithrix jacchus) in inhalation toxicity studies, its nasal morphology was examined. The nasal turbinates each consisted of one maxilloturbinate and one ethmoturbinate: these were more planar in structure than the comparable structures of rodents or dogs. The nasal cavity epithelia comprised squamous epithelium (SE), nasal transitional epithelium (NTE), respiratory epithelium (RE) and olfactory epithelium (OE), listed in order of occurrence from anterior to posterior positions. NTE was distributed as a narrow band lying between SE and RE. OE was limited to the dorsal part of the cavity, which was structurally similar to that of the macaque or man. Overall, this study revealed structural the similarity of the whole nasal cavity in the marmoset to that of macaques or humans. Prediction of nasal cavity changes in man based on extrapolation from experimentally induced changes in the common marmoset therefore seems likely to be feasible, making it a useful animal model for inhalation studies.

Animals↗

Parental care in the buffy-tufted-ear marmoset (Callithrix aurita) in wild and captive groups.

Studies on cooperative care of offspring in callitrichid primates are biased in favor of observations in captivity. In the wild, however, individuals have to deal with environmental pressures, which may influence their social behavior. We compared the individual effort attributed to parental care offered by members of a wild group (couple, plus a subadult helper) and two captive groups (A: couple, plus an subadult helper, B: couple, plus four adult helpers) of the buffy-tufted-ear marmoset, Callithrix aurita, from weeks 1-12 after the infants' birth. The carrier (breeding male and female or helper) and the infant's feeding (food sharing and foraging for food) were recorded. Up to week four, while the wild breeding pair shared infant carrying at similar proportions, the male from captive group A carried 100% of the time. Adult helpers from group B were the main carriers. Carrying behavior extended up to week 12 only in the wild group. Food provisioning to the infant was observed earlier in the groups wild and A, but general proportion of feeding records was lower in the wild than in captivity. Energetic cost of travelling and searching for food may be associated with equal division of carrying behavior by the wild breeding pair. Higher proportions of carrying in the groups wild and B may have delayed the development of the infants' motor skills required in foraging. Our data agree with previous studies: the father's lower investment in carrying when adult helpers are present and lower contribution of subadult non-reproductive members.

Animals↗

[Immune response produced by anti-rabies vaccines in marmosets (Callithrix sp)].

In order to evaluate the immune response produced by rabies vaccines in new world nonhuman primates, thirty marmosets (Callithrix sp) were divided into five groups of six individuals and submitted to five different antirabies vaccination schemes using two distinct commercially available animal vaccines. The first was produced in suckling mouse brain (Fuenzalida and Palacios), and the second in NIL-2 cell culture. Post-vaccine serological monitoring was carried out periodically. The results showed that the Fuenzalida and Palacios vaccine was not able to protect the animals when using a single dose or even with a booster. But when submitted to a vaccination routine similar to that used for humans, the marmosets showed detectable antibodies, and only one succumbed to rabies after being challenged. In addition, the vaccine produced in NIL-2 cell culture induced high antibody levels in all vaccinated animals and all animals survived the viral challenge.

Animals↗

The distribution of presumptive thoracic paraganglionic tissue in the common marmoset (Callithrix jacchus).

The aortic-pulmonary regions (APR) of seven adult marmosets (Callithrix jacchus) and the region of the right subclavian artery of a further three marmosets were diffusion-fixed with 10% buffered formol-saline solution. In both regions serial 5 microm sections were cut and stained by the Martius yellow, brilliant crystal scarlet and soluble blue method. Presumptive thoracic paraganglionic (PTP) tissue was only observed in the APR. PTP tissue was composed of small groups of cells that varied in size and number. The distribution of the groups of cells was extremely variable, so much so that it would be misleading to attempt to classify their position; they were not circumscribed by a connective tissue capsule, but were always related to the thoracic branches of the left vagus nerve. The cells lay in loose areolar tissue characteristic of this part of the mediastinum and received their blood supply from small adjacent connective tissue arterioles. Unlike the paraganglionic tissue found in the carotid body the cells in the thorax did not appear to have a profuse capillary blood supply. There was, however, a close cellular-neural relationship. The cells, 10-15 microm in diameter, were oval or rounded in appearance and possessed a central nucleus and clear cytoplasm. No evidence was found that these cells possessed a 'companion' cell reminiscent of the arrangement of type 1 and type 2 cells in the carotid body. In conclusion, we found evidence of presumed paraganglionic tissue in the APR of the marmoset which, however, did not show the characteristic histological features of the aortic body chemoreceptors that have been described in some non-primate mammals. A survey of the mediastina of other non-human primates is required to establish whether this finding is atypical for these animals.

Animals↗

Evidence of hemispheric specialization in marmosets (Callithrix penicillata) using tympanic membrane thermometry.

Recent studies have employed tympanic thermometry to assess lateralization of cognitive and emotional functions in primates. However, no studies using this technique have investigated the possibility of hemispheric specialization in New World monkeys. Therefore, the aim of the present study was to investigate tympanic membrane (TM) temperature asymmetries and their possible correlation with stress responses in marmosets (Callithrix penicillata). Infrared TM thermometry was completed bilaterally in 24 animals (14 males and 10 females) during a stressful situation of capture and restraint. There were no significant differences between gender. A significant negative correlation was observed between TM temperature of the right ear and the number of captures (r = -0.633; P<0.001). Subjects with a more frequent previous history of captures (5 to 9 captures; N = 11) showed lower TM temperature when compared to those with fewer previous captures (1 to 4 captures; N = 13). No differences were observed for the left TM temperature. These results suggest that under intense emotional challenge (capture and restraint) there is a stronger activation of the neural structures situated in the right brain hemisphere. Taken together, the data reveal for the first time evidence of hemispheric specialization in emotional physiological processing in a New World monkey.

Animals↗

Seasonality of the activity pattern of Callithrix penicillata (Primates, Callitrichidae) in the cerrado (scrub savanna vegetation).

Two wild groups of Callithrix penicillata, the Black Pincelled Marmoset, were observed from January to September 1998, in two areas, one an area of dense scrub savanna vegetation (cerrado) and the other, a semidecidual woodland (cerradão), both within the boundaries of the Ecological Reserve of IBGE (Brazilian Institute of Geography and Statistics), in an environmentally protected area, the APA (Portuguese abbreviation for "environmental protected area") Gama/Cabeça-de-Veado, Brasília, DF. The behavioral data collected during the rainy (January 15 to April 15) and dry season (June 1 to September 15) were compared. Because of the proximity to the Reserve facilities, the group from the dense scrub savanna vegetation (CD) was submitted to antropic impacts different from the group in the semidecidual woodland (CE), which was using as territory an area that had been suffering from man-made fires every two years as part of a long-term experimental project on fire impacts. The behavioral data was quantified by instantaneous cross-section ("scan sampling") every ten minutes with records of locomotion, rest, foraging for insects, use of exudate, and feeding. During the whole year, the greatest percentage of time spent by CE and CD was in foraging for insects, with 44% and 39%, respectively. It was evident when comparing the data for the two seasons that, for both groups, foraging for insects was more intense during the dry season, possibly to complement the shortage of food, and locomotion increased during the rainy season. The greater the availability and distribution of fruit in the areas, the greater the locomotion of the groups to obtain these resources. None of the other behavioral patterns, including the use of exudates, presented significant differences between the two seasons. Both groups foraged more frequently during the dry season and locomoted more during the rainy one.

Animals↗

Tal1/Scl gene transduction using a lentiviral vector stimulates highly efficient hematopoietic cell differentiation from common marmoset (Callithrix jacchus) embryonic stem cells.

The development of embryonic stem cell (ESC) therapies requires the establishment of efficient methods to differentiate ESCs into specific cell lineages. Here, we report the in vitro differentiation of common marmoset (CM) (Callithrix jacchus) ESCs into hematopoietic cells after exogenous gene transfer using vesicular stomatitis virus-glycoprotein-pseudotyped lentiviral vectors. We transduced hematopoietic genes, including tal1/scl, gata1, gata2, hoxB4, and lhx2, into CM ESCs. By immunochemical and morphological analyses, we demonstrated that overexpression of tal1/scl, but not the remaining genes, dramatically increased hematopoiesis of CM ESCs, resulting in multiple blood-cell lineages. Furthermore, flow cytometric analysis demonstrated that CD34, a hematopoietic stem/progenitor cell marker, was highly expressed in tal1/scl-overexpressing embryoid body cells. Similar results were obtained from three independent CM ESC lines. These results suggest that transduction of exogenous tal1/scl cDNA into ESCs is a promising method to induce the efficient differentiation of CM ESCs into hematopoietic stem/progenitor cells.

Animals↗

Serologic survey in a colony of captive common marmosets (Callithrix jacchus) after infection with herpes simplex type 1-like virus.

An outbreak of herpesvirus caused the death of four of five common marmosets (Callithrix jacchus) in a private colony. Gross lesions included acute ulcerative gingivitis, glossitis, and enlarged mandibular lymph nodes. Histologically, all fatal cases showed meningoencephalitis and eosinophilia with intranuclear inclusion bodies in neurons and glial cells. A herpes simplex-like virus was cultured from the brain and was identified as herpes simplex type 1 virus or a closely related virus by immunofluorescence. Serologic testing (complement fixation test) indicated that the surviving adult female was serologically positive for more than 4 yr and that the offspring she produced was seronegative. The most likely source of the outbreak was the owner who mouth fed hand-raised offspring.

Animals↗

Characterisation of the GH gene cluster in a new-world monkey, the marmoset (Callithrix jacchus).

In most mammals pituitary GH is encoded by a single gene with no close relatives. However, in man the GH gene has been shown to be one of a cluster of five closely related genes, four of which are expressed in the placenta. Rhesus monkey also expresses at least five closely related GH-like genes, although the genomic organisation of these has not been fully reported. Here we describe the cloning and characterisation of GH-like genes in a new-world monkey, the marmoset (Callithrix jacchus). This species possesses a cluster of eight GH-like 'genes'. The gene at the 5' end of this cluster encodes pituitary GH and is similar to that encoding human GH. Five of the eight marmoset 'genes' are probably pseudogenes, since they include mutations which would prevent normal expression, including stop codons and small insertions/deletions that would change the reading frame. In one case a large part of a gene is deleted, and in another a large insertion is introduced into an exon. The remaining two marmoset genes are potentially expressible, as proteins with sequences substantially different (at 25-30% of all residues) from that of marmoset GH itself; whether and in which tissue(s) such expression actually occurs is not yet known. None of the marmoset genes is clearly equivalent to any of the human GH-like genes expressed in the placenta, and this and phylogenetic analysis suggest that the duplications that gave rise to the marmoset GH gene cluster occurred independently of those that gave rise to the corresponding cluster in man. Although it includes more 'genes', the marmoset cluster extends over a shorter region of chromosomal DNA (about 35 kb) than does the human GH gene cluster (about 50 kb).

Amino Acid Sequence↗