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Tularemia in a common marmoset (Callithrix jacchus) diagnosed by 16S rRNA sequencing.

We report a case of tularemia in a common marmoset (Callithrix jacchus) diagnosed by determination of the isolate's 16S ribosomal RNA (rRNA) gene sequence. Pathological examination of the animal revealed a multifocal acute necrotizing hepatitis, interstitial nephritis, splenitis, and lymphangitis of the mandibular, retropharyngeal, and cervical and mesenteric lymph nodes. Moreover, multiple foci of acute necrosis were found in the epithelium of the jejunum and the interstitium of the lung. Bacteriological investigations revealed a septicemia. The isolated infectious agent was uncommon, not routinely diagnosed in our laboratory and therefore difficult to identify by conventional tools in a reasonable time and effort. thus, we decided to perform a genetic analysis based on the 16S rRNA gene sequence. Thereby, an infection with Francisella tularensis, the causative agent of tularemia, was unambiguously diagnosed. This shows the great advantage 16S rRNA gene sequencing has as a general identification approach for unusual or rare isolates.

Animals↗

Artificial insemination in Callithrix jacchus using fresh or cryopreserved sperm.

Assisted reproductive techniques are needed urgently to facilitate the captive breeding of many New World primate species which are endangered in the wild and to assist the effective genetic management of small colonies. A protocol was devised for artificial insemination in the common marmoset, Callithrix jacchus, using ejaculated sperm obtained by vaginal washing after copulation. A double insemination protocol was employed, with the first insemination taking place the day before ovulation was expected to occur and the second 48 h later. All six females inseminated with fresh ejaculated sperm became pregnant, delivering a total of 16 offspring at term. The gestation lengths and litter sizes were not statistically different from those observed in pregnancies following natural mating. The insemination protocol was adapted for use with cryopreserved ejaculated sperm by including an additional insemination on the day of expected ovulation, to take into account differences in the capacitation time of frozen-thawed sperm compared to fresh sperm. Three out of six females inseminated according to this triple insemination schedule, conceived, although one female subsequently resorbed twin foetuses approximately 100 days later. The remaining two pregnant females delivered four babies at term, one singleton and one set of triplets. In the final group, six females were inseminated with low doses of cryopreserved epididymal sperm using the same triple insemination protocol used for frozen-thawed ejaculated sperm. One female conceived, delivering triplets.

Animals↗

Inducible nitric oxide synthase (iNOS) expression in liver and splenic T lymphocyte rise are associated with liver histological damage during experimental hepatitis A virus (HAV) infection in Callithrix jacchus.

Callithrix jacchus is considered a reliable animal model for hepatitis A virus (HAV) infection. All three HAV orally inoculated marmosets developed hepatitis - the infection was monitored by continuous virus shedding, high levels of serum enzyme alanine aminotransferase, specific antibody and seroconversion 3-6 weeks after HAV inoculation. HAV antigen was detected in liver by immunofluorescence 4 days post inoculation (PI) and onwards. To gain insight into the biological role of inducible nitric oxide synthase (iNOS) during immune-related acute liver injury the enzyme was searched in frozen biopsies: immunofluorescent labeling was found in the cytoplasm of liver cells mainly Kupffer's cells and spleen macrophages (CD68+) starting 11 days PI with maximum intensity on the fifth to sixth week PI. Necroinflammatory liver lesions characteristic of viral hepatitis were also observed at 10 days PI with maximum severity at 4 to 6 weeks PI. Furthermore, T lymphocytes (CD2+) were raised at this time point. No difference was evident in the frequency of B lymphocytes (CD20+). Therefore, iNOS expression preceded necroinflammatory liver lesion and maximal immunofluorescence reaction was coincident with tissue injury, supporting the hypothesis that NO contributes to hepatic cytotoxic mechanism but also to virus clearance. The concomitant rise in T-lymphocyte population may suggest a role for these cells in this and/or other independent HAV-induced pathological changes.

Animals↗

Histopathological and immunohistochemical studies of hepatitis A virus infection in marmoset Callithrix jacchus.

Samples of serum, feces and liver tissue and organs of six cotton-eared marmosets Callithrix jacchus infected intravenously with two different strains of hepatitis A virus (HAV), were studied by conventional histologic techniques, by serological techniques and by immunocytochemical methods, such as immunofluorescence (IF) and peroxidase-antibody techniques. Hepatitis A antigen (HAAg) was detectable in daily collected stools, in liver biopsy obtained sequentially, and in organs collected at necropsy. Two marmosets also developed antibodies to HAV. By contrast, serum transaminases were not altered and there were histological hepatic lesions consistent with acute viral hepatitis in all inoculated animals. The data obtained, demonstrate that these primates are susceptible to human HAV and may be a useful animal model for the study of infection by this virus.

Animals↗

Morphological characteristics of dopaminergic immunoreactive neurons in the olfactory bulb of the common marmoset monkey (Callithrix jacchus).

The present study describes the distribution of tyrosine hydroxylase (TH)-immunoreactive (IR) elements in the olfactory bulb of the common marmoset monkey (Callithrix jacchus), a primate species by immunohistochemistry. We identified six layers of the olfactory bulb of the common marmoset monkey in sections stained with cresyl violet. The majority of TH-IR cells were found in the glomerular layer. A few TH-IR cells were present in the external plexiform and granule cell layers. TH-IR fibers were identified in all layers of the olfactory bulb. The density of these nerve fibers was high in the internal plexiform and granule cell layers. The results in the olfactory bulb of the common marmoset monkey are generally similar to previous reports in some mammals. These data suggest that TH in the olfactory bulb of the common marmoset monkey may play a role in olfactory transmission via the glomeruli like in other mammals.

Animals↗

Morphology of thyroid follicular cells from adult marmosets (Callithrix jacchus).

The existence of follicles with a "super flat" epithelium and the large number of primary and especially secondary lysosomes and lipofuscin granules in these cells are characteristic features of the thyroid gland of adult marmosets (Callithrix jacchus, new-world monkeys). It is supposed that these morphological findings are indications of a relatively low function and early ageing.

Animals↗

C-cells of marmosets (Callithrix jacchus).

The localization and fine structure of C-cells of the thyroid glands from 31 marmosets (Callithrix jacchus) of different age groups (newborn to 11 years old) were investigated by electron microscopy and immunomorphology (APAAP, calcitonin). Calcitonin-positive cells were found concentrated in the middle third of the gland. Their localization in the follicular wall or in the interstitial space could not clearly be demonstrated by light microscopy. In adult animals, they were always located within the basal lamina, as revealed by electron microscopy. In newborn animals, their localization was difficult because of the dense cell packing. The fine structure of the C-cells with their 200 nm electron-dense granules corresponded to that of other species. Two cell types could be distinguished: (1) granule-rich cells in the storing phase and (2) cells with fewer granules and well-developed rough endoplasmic reticulum in the synthesis phase. The concentration of C-cells in the middle third of the marmoset thyroid gland renders this species especially suitable for morphological investigations and in vitro experiments.

Aging↗

Stereology of the myocardium in two species of Callithrix (Callitrichidae, primates).

The majority of studies on cardiac morphology have concentrated on Old World monkeys. Ten marmoset hearts of the genus Callithrix were studied (5 hearts of C. jacchus and 5 of C. penicillata), dissected and fixed in a 10% buffered formaldehyde solution, pH 7.2. Unbiased stereological estimates were obtained from isotropic uniform random sections of the myocardium. For stereological quantification the myocardium was regarded as consisting of cardiac myocytes and interstitium. The volume density (Vv) was determined by point counting. We used the disector method to obtain the numerical density of the cardiac myocytes (Nv[nuclei]). Myocardial stereological differences between the two species of marmoset were not statistically significant. We can therefore determine the pooled Vv[myocyte] and Nv[nuclei] as 68.6% and 41.6% (10(3)/mm3) respectively. The values found for Vv[myocyte] and Nv[nuclei] in the marmoset are respectively about 23.0 and 92.0% greater than those of the baboon, and respectively 57.3 and 45.5% greater than those in man. In contrast, the mean myocyte volume in the marmoset is not significantly different to that of man but is almost 36.0% less than that of the baboon.

Animals↗

Investigation of the sleep electrocorticogram of the common marmoset (Callithrix jacchus) using radiotelemetry.

OBJECTIVE: To evaluate the use of a totally implantable radiotelemetry system for recording the sleep electrocorticogram (ECoG) of a small new world primate, the common marmoset (Callithrix jacchus) without restraint during data collection. METHODS: Under anaesthesia a telemetry transmitter, which allowed the recording of a single ECoG channel, was implanted intraperitoneally. This system allowed ECoG data to be recorded overnight from animals living in pairs within their habitual laboratory environment over a period of 12 months. Data were subsequently scored using modified Rechtschaffen and Kales criteria (A manual of standardized terminology, techniques and scoring system for sleep stages of human subjects. Los Angeles, UCLA Brain Information Service/Brain Research Institute, 1968) into stages of waking, light sleep, deep sleep and probable rapid eye movement sleep (pREM). Concurrent video recording was used to assist in the categorising of pREM. RESULTS: Results showed that, as in man, the marmoset exhibits sleep cycles with stages alternating between non-REM (deep sleep and light sleep) and pREM sleep throughout the night. In common with other non-human primates the duration of each of the sleep stages was relatively short and punctuated with frequent waking. CONCLUSIONS: These data suggest that sleep in marmosets housed under laboratory conditions (a) can be recorded without restraint and (b) has potential to be used as a model for human sleep.

Animals↗

Plasmodium brasilianum in the common marmoset Callithrix jacchus.

Chronic quartan malarial infection has been established in the common marmoset (Callithrix jacchus). Plasmodium brasilianum from a douroucouli monkey (Aotus trivirgatus) was used to infect splenectomized twin animals, passed to an intact animal, and then to 4 other intact adults, 2 pairs of twins. In 2 of the 4 latter animals there was continuing patency with parasitaemias of less than or equal to 0.5% parasitized erythrocytes for 30 weeks. The other 2 had lower initial levels of parasitaemia; in 1 of these parasitaemias remained low or subpatent. All marmosets developed lymphocytosis. One animal became ill 30 weeks after infection with anaemia, weight loss and mild proteinurea, the other 3 remained well. Histological examination showed minor changes in the kidneys; spleens of infected animals showed marked follicular hyperplasia and phagocytosis of pigment. The livers showed sinusoidal hypercellularity and pigment deposition and in splenectomized animals, a marked lymphoid follicular hyperplasia in the portal tracts.

Animals↗

Plasmodium vivax malaria in the common marmoset, Callithrix jacchus: adaptation and host response to infection.

Infection with Plasmodium vivax was established in splenectomized Callithrix jacchus marmosets by inoculation of parasitized blood from Aotus trivirgatus carrying the Vietnam Palo-Alto line of P. vivax. Subsequent blood passage through intact marmosets resulted in higher peak parasitaemias (about 1% of red cells infected) and the loss of stainable Schüffner's dots in infected cells. Primary infections with the adapted line were patent for 74 days or more, and induced both a substantial antibody response, as determined by indirect fluorescence, and some lymphocytosis, but no marked anaemia. Marmosets which had recovered from their primary infection (or in which it was drug-cured) suffered abbreviated patency with low-grade parasitaemia on re-infection.

Animals↗

Functional architecture of area 17 in normal and monocularly deprived marmosets (Callithrix jacchus).

The organization of the primary visual cortex (VI) of the common marmoset (Callithrix jacchus) was studied both physiologically and by means of transneuronal labelling of geniculocortical afferents. We addressed the question whether monocular deprivation (MD) could stabilize segregation into ocular dominance (OD) columns, which are not seen in normal adult marmosets but are present in juvenile animals (Spatz, 1979, 1989). Properties of neurons in normal marmosets closely resembled those of other New-World and Old-World monkeys and orderly tangential progressions of preferred orientation were observed. However, in contrast to species that display well-defined OD columns, neurons of layer 4 in V1 of normal adult marmosets received balanced inputs from the two eyes. Early MD (even though followed by prolonged binocular experience into adulthood) resulted in a reduction of cell size in laminae of the lateral geniculate nucleus with input from the deprived eye and a dramatic overall shift in ocular dominance towards the nondeprived eye in the cortex. However, isolated clusters of cells dominated by the deprived eye were found in both layers 4 and 6. Injection of lectin-conjugated horseradish peroxidase (WGA-HRP) into the deprived eye revealed elongated patches of terminal label, about 350 microns wide, in flat-mounted sections through layer 4. Afferent segregation was sharper and more regular in the region of V1 representing parafoveal visual space than in that representing the fovea. Our findings support the notion that all Old-World and New-World monkeys possess the capacity for segregation of geniculocortical afferents into OD columns.

Afferent Pathways↗

Topography of ganglion cells and photoreceptors in the retina of a New World monkey: the marmoset Callithrix jacchus.

We studied the anatomical substrates of spatial vision in a New World monkey, the marmoset Callithrix jacchus. This species has good visual acuity and a foveal specialization which is qualitatively similar to that of humans and other Old World primates. We measured the spatial density of retinal ganglion cells and photoreceptors, and calculated the relative numbers of these cell populations. We find that ganglion cells outnumber photoreceptors by between 2.4:1 and 4.2:1 in the fovea. The peak sampling density of ganglion cells is close to 550,000 cells/mm2. This value falls by almost 1000-fold between the fovea and peripheral retina; a value which approaches recent estimates of the centroperipheral ganglion cell gradient for human and macaque monkey retina and primary visual cortex. The marmoset shows a sex-linked polymorphism of color vision: all male and some female marmosets are dichromats. Six of the retinas used in the present study came from animals whose chromatic phenotype was identified in electrophysiological experiments and confirmed by polymerase chain reaction (PCR) amplification of cone opsin encoding genes. One animal was a trichromat and the others were dichromats. Antibodies against short wavelength-sensitive (SWS) cones labeled close to 8% of all cones near the fovea of one dichromat animal, consistent with electrophysiological evidence that the SWS system is present in all marmosets. The topography and spatial density of cone photoreceptors and ganglion cells was similar to that reported for macaque retina, and we found no obvious difference between dichromatic and trichromatic marmoset retinas. These results reinforce the view that the main determinate of primate foveal topography is the requirement for maximal spatial resolution.

Animals↗

The morphology and distribution of horizontal cells in the retina of a New World monkey, the marmoset Callithrix jacchus: a comparison with macaque monkey.

The morphology and distribution of horizontal cells was studied in the retina of a New World monkey, the marmoset, Callithrix jacchus, and compared with that of the Old World macaque monkey. Horizontal cells in macaque and marmoset were either labelled with the carbocyanine dye, DiI, and then photoconverted, or were labelled by intracellular injection with Neurobiotin. The marmoset has two types of horizontal cell, H1 and H2, which have dendritic and axonal morphology similar to their counterparts in Old World monkeys and human. The dendritic-field size of both cell types increases with distance from the fovca. Both types make contact with the vast majority of the cones within their dendritic field. The dendrites of H1 cells in marmoset contact almost twice as many cones as H1 cells in macaque at an equivalent eccentricity. With increasing distance from the fovea, H1 cells make contact with more cones but have, on average, fewer terminal knobs inserted in each cone. The increase in dendritic-field area of H1 cells is balanced by a decrease in spatial density (from 4500 cells/mm2 at 25 deg eccentricity to 1000 cells/mm2 in far peripheral retina), so coverage of the retina remains fairly constant, between 5 and 8. Overall, the results show that the qualitative morphological properties, as well as quantitative population properties of horizontal cells, are common to both New World and Old World primates.

Animals↗

Effect of monocular deprivation on NMDAR1 immunostaining in ocular dominance columns of the marmoset Callithrix jacchus.

We previously showed that immunoreactivity to N-Methyl-D-aspartate (NMDA) receptors in primary visual cortex of Callithrix jacchus is regulated by visual activity during the second and third postnatal months (Fonta et al., 1997). The purpose of the present study was to show that the columnar pattern of high and low NMDAR1 immunoreactivity observed in monocularly deprived animals corresponds to ocular dominance columns linked to the nondeprived and deprived eye, respectively. We compared cortical distribution of NMDAR1 receptors and the projection zones of thalamic afferents, revealed by transneuronal transport of tritiated proline, in 2-month-old, either monocularly deprived or control, marmosets. The data show that ocular dominance columns exist in 2-month-old marmosets and that a 2-week monocular deprivation by means of eyelid suture leads to a modification of the thalamo-cortical afferents organization. Experiments of neuronal tracing and immunohistochemistry performed on the same animals demonstrated that cortical domains with decreased NMDAR1 level correspond to the deprived eye columns. These investigations, coupled to the previous results, strongly suggest that the NMDA receptors, regulated by visual activity, are involved in the refining of ocular dominance columns in the primary visual cortex of juvenile marmoset.

Animals↗

Long-range interactions in the lateral geniculate nucleus of the New-World monkey, Callithrix jacchus.

Visual stimulation of zones extending beyond the classical receptive field can modulate the contrast gain of neurons in the lateral geniculate nucleus (LGN) of cats, but little is known about the effect of extra-classical visual stimulation on the LGN of primates. Hence, we compare the effect of long-range interactions in parvocellular and magnocellular LGN layers of the marmoset monkey Callithrix jacchus using optimal, incremental spots flashed on the classical receptive field either alone or simultaneously with the shift of a grating (98% contrast; 0.1 cycles/deg) confined to a peripheral annulus (radii: 5-15 deg). The contrast required to drive the response halfway to saturation (c50) of most LGN neurons was raised by remote pattern shifts. The c50 ratio [(shift+spot)/spot] in OFF-center magnocellular neurons was significantly higher than in OFF-center parvocellular neurons. OFF-center magnocellular neurons closer to the fovea (<10 deg eccentricity) tended to have a higher c50 ratio than in more peripheral neurons. A significant drop in visual sensitivity to 25% contrast spots was observed during remote motion: d' fell from 1.8 to 1.4 in parvocellular neurons and from 2.2 to 1.7 in magnocellular neurons. Such long-range interactions produce a reduction in visual sensitivity by changing the gain of the geniculate relay and point to an inhibitory, motion-sensitive extra-classical receptive field in both parvocellular and magnocellular pathways, which may be involved in saccadic suppression and attentional mechanisms in early vision.

Animals↗

The distribution of calcium-binding proteins in the lateral geniculate nucleus and visual cortex of a New World monkey, the marmoset, Callithrix jacchus.

Antibodies directed against the calcium-binding proteins, parvalbumin and calbindin, can be used to label distinct neuronal subgroups in the primate visual pathway. We analyzed parvalbumin immunoreactivity (P-IR) and calbindin immunoreactivity (C-IR) in the lateral geniculate nucleus (LGN) and visual cortex of the marmoset, Callithrix jacchus. We compared marmosets which were identified as having dichromatic or trichromatic color vision. Within the LGN, the density of P-IR neurones is highest in the parvocellular and magnocellular laminae, but C-IR neurones are found mainly in the koniocellular division of the LGN, that is, the interlaminar zones and S laminae. Not all interlaminar zone cells are C-IR. In the visual cortex, P-IR neurones are present in all laminae except lamina 1, in areas V1 and V2. Neurones which are strongly C-IR are mainly located in laminae 2 and 3 in V1 and V2. Lightly C-IR neurones are concentrated in lamina 4, and are more numerous in V1 than in V2. Quantitative analysis showed no differences in the density or distribution of IR neurones in either LGN or visual cortex when dichromat and trichromat animals were compared. We conclude that this functional difference is not associated with differences in the neurochemistry of calcium-binding proteins in the primary visual pathways.

Animals↗

Chemical ecological characteristics of herbivory of Siparuna guianensis seeds by buffy-headed marmosets (Callithrix flaviceps) in the Atlantic Forest of southeastern Brazil.

The buffy-headed marmoset (Callithrix flaviceps) is apparently the only predispersal herbivore of the seeds of Siparuna guianensis at the Caratinga Biological Station in southeastern Brazil. Both the fruit receptacles and the frutioles (seeds) of S. guianensis are relatively rich in nutrients such as carbohydrates, proteins, and lipids, but the receptacles contain high concentrations of benzylisoquinoline alkaloids. The latter presumably act as a qualitative chemical defense, impeding the access of potential predators to the alkaloid-poor frutioles. However, on ripening, the receptacle splits open, exposing the frutioles, which enables C. flaviceps to avoid the plant's chemical defenses effectively. Taking care to avoid contact with the receptacle, the marmosets pluck out the frutioles and ingest the seeds. Qualitative and quantitative changes (in particular a significant reduction in daily ranging) in the marmosets' behavior during the period when S. guianensis frutioles were accessible indicate that this was a preferred plant resource. This is the first record of the consumption of S. guianensis seeds by callitrichine monkeys (which are not known to be systematic seed eaters), despite the fact that both are widely distributed in the Neotropics. It is thus possible that the behavioral strategy observed here is a unique phenomenon resulting from a specific combination of factors, including the abundance of S. guianensis within the study area. The lack of other records may nevertheless be a result of insufficient sampling effort, in which case, the feeding strategies of callitrichines may have an important influence on the reproductive biology of S. guianensis in many areas.

Alkaloids↗