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XY female marmoset (Callithrix jacchus).

A marmoset (Callithrix jacchus) with atypical external genitalia was phenotypically and genetically characterized. Testosterone concentration correlated with that of female marmosets. Externally, there was only one opening for the urethra. Internal genitalia were characteristic of those of female marmosets, and consisted of ovaries, with follicles in various developmental stages, and uterus. Microscopically, a normal vaginal structure was found. An XX/XY chimerism and high steroid hormone values are normally found in common marmosets. Genetic analysis was used for in vivo determination of sex. The Y-linked zinc finger protein gene (ZFY) last intron, and sex-determining region Y gene (SRY) exon were found by use of polymerase chain reaction and posterior sequencing analyses, indicating that this marmoset had Y-linked chromosome sequences. Normal SRY exons can, therefore, be associated with female internal sexual organs in marmosets; this may be the first XY female described in non-human primates.

Animals↗

Cytologic assessment of the vaginal epithelium in the common marmoset (Callithrix jacchus): a preliminary new approach to reproductive screening.

The application of vaginal cytology in defining reproductive events is a widely accepted and applied diagnostic tool in many species. Here we present a preliminary approach to recognizing a correlation between changes in vaginal epithelial cytology and circulating plasma progesterone in the common marmoset, Callithrix jacchus, such that vaginal cytology could be diagnostically appropriate for reproductive management. We assessed six captive-born adult female common marmosets for the duration of one complete estrus cycle. Cytologic samples, collected by vaginal swab, and blood for plasma progesterone concentration were collected twice weekly. Cytologic samples were blindly assessed, and vaginal epithelial maturation was defined on the basis of the karyopyknotic index (KPI). In the three consistently cycling females (cj0328, cj0732, and cj0678), a marked increase in KPI corresponded with rising plasma progesterone levels, which subsequently decreased throughout the duration of follicular phase. The remaining three females who had previously irregular estrus cycles (cj0608, cj0680, cj0724) demonstrated irregularity in their progesterone profiles which correlated with inconsistent KPIs. Although KPI is not a very sensitive single sampling indicator of ovulation, evaluating changes in cytologic morphology over time aid in predicting the timing of an ovulatory event. Our results suggest that vaginal cytology can be an effective alternative to blood sampling for reproductive management of the common marmoset.

Animals↗

Calcinosis circumscripta in a common marmoset (Callithrix jacchus jacchus).

A 2.5-y-old, male common marmoset (Callithrix jacchus jacchus) developed a 2-cm, interscapular, subcutaneous mass with variably firm and cystic areas. Radiographs demonstrated a radiodense mass in close proximity to a previously implanted microchip. Fine-needle aspiration yielded a chalky liquid that, on cytologic examination, contained amorphous debris. Aerobic and anaerobic cultures were negative. Surgical excision required extensive dissection, with the mass infiltrating deep to the scapula and extending to the mammary gland. The mass weighed 30 g and comprised 10% of the animal's body weight. Microscopic examination demonstrated multifocal, variably sized, amorphous aggregates of granular, basophilic material (mineral) in the subcutis and extending to skeletal muscle. Mineral deposits were surrounded by macrophages, giant cells, and fibrous connective tissue. A focal area of ectopic bone production was present. Crystallographic analysis and x-ray diffractometry determined the material to be comprised of 100% hydroxyapatite. These findings were consistent with a diagnosis of calcinosis circumscripta. Systemic metabolic abnormalities were excluded based on examination of complete blood count, serum chemistry, and ionized calcium. Calcinosis circumscripta in the common marmoset has not previously been reported, although the lesion has been reported to occur in rhesus macaques and is well described in man and dogs. Accumulation of calcium deposits and production of ectopic bone in a marmoset is interesting in light of this species's unique calcium and vitamin D metabolism.

Animals↗

Ontogeny of monooxygenase activities in the marmoset monkey (Callithrix jacchus).

The pre- and postnatal development of monooxygenases in the liver and adrenal gland of marmoset monkeys (Callithrix jacchus) was investigated. Cytochrome P450 was detected in the fetal adrenal gland, but aldrin epoxidase, ethoxycoumarin O-deethylase, and ethoxyresorufin O-deethylase activities were below detection limits. Although fetal hepatic cytochrome P450 was not detected, low activities of aldrin epoxidase and ethoxycoumarin O-deethylase, but no ethoxyresorufin O-deethylase, could be detected in fetal liver. These enzymes attained adult marmosets activities when the offspring were approximately 2 months of age.

7-Alkoxycoumarin O-Dealkylase↗

Collection and analysis of semen from the common marmoset (Callithrix jacchus).

The authors examined the seminal characteristics of 16 male common marmosets (Callithrix jacchus) to provide baseline data for future studies of the reproductive biology of this species. Semen samples were collected by electroejaculation. There was considerable inter- and intra-male variation in all seminal characteristics. The median seminal volume was 30 microliters (range 8 to 85 microliters), and the median total sperm count was 5.1 x 10(6) sperm (range, 0.1 to 43 x 10(6]. The median progressive sperm motility was 48% (range, 10% to 76%), and 49% of the sperm exhibited normal morphology (range, 24% to 81%). Three types of sperm head abnormalities and eight types of tail defects were noted. Tail defects were common (median, 50%; range, 17% to 76%), whereas head defects were relatively rare (median, 4.5%; range, 0% to 24%). The results indicate that semen samples can be routinely collected from this species, but considerable inter- and intra-male variation can be expected. It is therefore important to examine several semen samples from each male marmoset to obtain an accurate seminal picture.

Aging↗

Plasma vitamins A and E and hydrogen peroxide-induced in vitro erythrocyte haemolysis in common marmosets (Callithrix jacchus).

Plasma total lipid, cholesterol, all-trans retinol (vitamin A) and alpha-tocopherol (vitamin E) concentrations, and the susceptibility of erythrocytes to hydrogen peroxide-induced haemolysis in vitro were investigated in healthy laboratory-bred common marmosets (Callithrix jacchus). The concentrations of alpha-tocopherol and total cholesterol were similar to those of control human subjects. The mean lipid concentration was higher (P less than 0.025) and the retinol concentration lower (P less than 0.001) in the marmosets. The susceptibility of the erythrocytes of the marmosets to hydrogen peroxide-induced haemolysis was high, both in absolute value and relative to the controls. Changes in phospholipid and fatty acid composition might have rendered the red cells susceptible to oxidative stress.

Adult↗

Connective tissue of the livers of newborn and adult marmosets (Callithrix jacchus).

Immunofluorescence microscopic and electron microscopic investigations revealed components of the matrix and of the basal lamina (collagen type I, III, IV and V, BL-heparan sulfate and fibronectin) in the sinus wall (Disse's space) of the livers of newborn and adult marmosets (Callithrix jacchus). Collagen type I was missing in both the two age groups. Small amounts of laminin were present in the livers of newborn and absent in those of adult animals, whereas collagen type III occurred in the form of delicate fibres. Light microscopic inspection showed a continuous distribution of all other components in the sinus wall. The amount of collagen type III and V increased depending on the age. Electron microscopic investigations revealed single or bundled fibrils (20-30 nm) and filaments (10-12 nm). After addition of tannic acid, plaques of a fine-filamentous network and incorporated granules were observed. After addition of resting Ruthenium Red, electron-dense granules (20-60 nm) were irregularly distributed in the structureless space, resting on collagenous fibrils and cell membranes. The fibrils were allocated to collagen type III, the filaments to collagen type V. The plaques were supposed to contain heparan sulfate, collagen type IV and fibronectin. The absence of a Lamina densa of the basal lamina was attributed to the absence of laminin which probably plays an important role in the formation of this layer. Differences in the distribution pattern of the matrix components and thus a functional mosaic of the permeability of Disse's space were assumed. The complete absence of collagen type I and laminin in the lobules makes the adult marmoset liver especially suited for studies on the importance of this collagen type under pathological conditions, since both components are expressed in this way.

Animals↗

Lymphocyte subsets in the young and aging marmoset (Callithrix jacchus).

T lymphocytes were studied in healthy young and old marmosets (Callithrix jacchus). Data indicate that monoclonal antisera to human T lymphocyte antigens can be used to identify these cells in marmosets. Total T and suppressor T cells were lower in the older group; there were no differences in helper T or in B cells. Helper/suppressor ratios (T4/T8) were increased in the older group. Thus, immune system alternations with aging and/or immune deficiency syndromes may be studied in this animal model using available reagents.

Aging↗

Determination of ovulation and pregnancy in the marmoset (Callithrix jacchus) by monitoring of urinary hydroxypregnanolone excretion.

In Callithrix jacchus the urinary excretion of hydroxypregnanolone (5 alpha-pregnane-3 alpha, 7 alpha-diol-20-one) shows a luteal rise during the ovulatory cycle. This progesterone metabolite can therefore be used as an indication of ovulation, implantation, and subsequently for the persistence of the pregnancy. The excretion can be monitored by high performance thin-layer chromatography (HPTLC) and postchromatographic derivatization after enzymatic hydrolysis of the conjugate. Profiles of urinary hydroxypregnanolone levels are presented and correlated with luteinizing hormone excretion. The method is noninvasive and therefore suited for long-term studies in these monkeys.

Animals↗

Identification of the main gestagen metabolite in marmoset (Callithrix jacchus) urine by NMR and MS spectroscopy.

Hydroxypregnanolone is the most abundant progesterone metabolite in the urine of marmoset monkeys (Callithrix jacchus). The substance is excreted as conjugate. The concentration of this steroid may be monitored by high performance, thin layer chromatography and postchromatographic derivatization. Hydroxypregnanolone was purified and subsequently identified by NMR spectroscopy and gas chromatography/mass spectroscopy. The exact chemical structure is 5 alpha-pregnane-3 alpha, 7 alpha-diol-20-one.

Animals↗

Endogenous N-excretion and minimal protein requirement for maintenance of the common marmoset (Callithrix jacchus).

The protein requirement for maintenance of the marmoset Callithrix jacchus was investigated by factorial calculation via the N balance technique. Six adult male marmosets were fed a protein-free pelleted diet for 22 days. When a low protein diet (6%) was fed in the preperiod, only 7 days were necessary and when fed a higher protein diet (18%) 10 days were necessary to reach a minimum of N excretion during protein depletion. Body weight losses averaged 11% and food intake was 21% lower in the protein free period. The average endogenous urine N (EUN) was 62.1 mg N and endogenous fecal N (EFN) 49.2 mg N/kg W0.75. Body surface losses (BSN) were assumed to be 20 mg N/kg W0.75. The total endogenous loss of 131.3 mg N/kg W0.75 is similar to that of other species. Net protein utilization (NPU) of well balanced proteins, determined near the N equilibrium, was 51.8%. The calculated net protein requirement amounts to 313.6 mg N/kg W0.75, which corresponds to a dietary protein content of 6% in a diet of high caloric density, (about 19-20 MJ/kg dry matter) when the food intake is 3.3% of body weight.

Animal Nutritional Physiological Phenomena↗

Refractoriness of Callithrix penicillata red blood cells to Plasmodium falciparum in vivo and in vitro.

Attempts to infect the New World marmot Callithrix penicillata with Plasmodium falciparum were unsuccessful. Attempts were also made to infect red blood cells of C. penicillata and Saimiri sciureus with P. falciparum in vitro, and these too were unsuccessful due to a high rate of hemolysis produced by apparently adverse culture conditions. It is concluded that modifications to the existing culture conditions will need to be made before successful parasitemia can be induced in vitro in simian erythrocytes.

4-Aminobenzoic Acid↗

Experimental infections of simians with human malaria: attempts to infect Callithrix penicillata with Plasmodium falciparum.

After reviewing the use of non-human primates of the Old and New Worlds for human malaria research, we concluded that another experimental animal which is easily available to use and possible to rear indoors is needed. Thus, we studied the susceptibility of the marmoset Callithrix penicillata to Plasmodium falciparum erythrocytic infections. The marmosets received various P. falciparum human isolates, directly from a patient and from continuous cultures. The Palo Alto strain, which has been adapted to the night monkey Aotus trivirgatus and further maintained in the squirrel monkey Saimiri sciureus was also used. In a total of 20 marmosets we performed 31 inoculations, with 10(5) to 10(9) parasites, intraperitoneally, intracardiacly or intravenously. Blood samples from each animal were examined daily up to day 90 post-inoculation. None of the intact marmosets developed patent infections. Four out of 19 C. penicillata, previously splenectomized, showed circulating parasites for up to five days after intravenous inoculation with the Palo Alto strain, becoming negative thereafter. Neither the addition to the simian diet of p-aminobenzoic acid, essential for the parasite metabolism, nor drug-immunosuppression, improved the marmoset susceptibility to P. falciparum.

Animals↗

Presence of pH2-sensitive circulating interferon among Callithrix jacchus marmosets.

Significant and relatively stable levels of serum-interferon were demonstrated in a Callithrix jacchus population. This circulating interferon was acid-sensitive in all cases, classifying it as immune or "gamma-type" interferon. Our results in these hematopoietic chimeras suggest that the presence of immune-type or at least pH 2-sensitive interferon could be related to the presence of two allogenic lymphocyte populations in each marmoset.

Animals↗

The postnatal development of the temporomandibular joint of the common marmoset (Callithrix jacchus).

The temporomandibular joints (TMJ) of 55 male and 50 female common marmosets (Callithrix jacchus) with an age range of 1 day- to < 12-yr-old have been studied to extend the knowledge on the craniomandibular articulation of this primate. Details of changes in the bone structure, gross and functional anatomy, and histologic appearance of the marmoset TMJ during the neonatal period, infancy, childhood, adolescence, and adulthood are compared with those which occur in the TMJ of man. It is concluded that further investigations on the structure and function of the marmoset TMJ and jaws may establish this nonhuman primate as a valuable analog for TMJ research in man.

Aging↗

Skin reaction in Callithrix jacchus penicillata to phytohemagglutinin.

Sixty marmosets (Callithrix jacchus penicillata) were intradermally tested with phytohemagglutinin. It was possible to classify this population into three groups on the basis of the reactions: 1. negatives, 2. those which showed just local hemorrhage, and finally 3. a group which developed a wheal as well as hemorrhage. The biopsy at the site of the reaction demonstrated several microscopic foci. This finding contrasted with the homogeneous cellular infiltration found in the human controls.

Animals↗

Poxvirus infection in a colony of common marmosets (Callithrix jacchus).

An epizootic poxvirus infection occurred in a colony of 80 common marmosets (Callithrix jacchus) recently introduced to a laboratory facility. Over an 18-week period, 29 of the monkeys exhibited skin lesions that persisted for 4-6 weeks. Although eight marmosets died during the outbreak, their deaths were not attributed directly to the poxvirus infection. The skin lesions developed over the entire body surface including the soles and palms. Initially characterized as erythematous papules, they quickly changed to elevated coalescing lesions with extensive scab formation. Histopathologically, the lesions revealed moderate to marked acanthosis, and they progressed to full-thickness epidermal necrosis and ulceration. Intracytoplasmic inclusion bodies were observed occasionally within degenerate keratinocytes. These inclusions most probably constituted the intracytoplasmic aggregates of viral particles observed ultrastructurally and confirmed as members of the poxvirus group by negative staining of direct skin scrapings.

Animals↗

The structure and epididymal maturation of the spermatozoon of the common marmoset (Callithrix jacchus).

Maturation of the spermatozoon of the common marmoset (Callithrix jacchus) during epididymal transit was investigated using light and electron microscopy. Except for the caudal migration of the cytoplasmic droplet there were no apparent ultrastructural modifications, although interesting morphological features of the sperm head were observed such as the substructure of the acrosome and a waist at the equatorial region. Alteration to the plasmalemma during maturation was reflected by an increasing capacity of the sperm surface to bind wheat germ agglutinin (recognising n-acetyl-d-glucosamine) but not ricin communis or concanavalin agglutinins. This change was concomitant with the development of sperm fertilising capacity in this species.

Acrosome↗