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A survey of the pathology of marmosets (Callithrix jacchus) derived from a marmoset breeding unit.

The results of a survey of the major pathological conditions encountered in an established breeding colony of common cotton-eared marmosets (Callithrix jacchus) is presented. 265 home-bred and 70 imported wild-caught marmosets were examined. A Heinz body haemolytic anaemia and skeletal muscle myopathy were the most common pathological findings and were considered to be a result of a complex nutritional deficiency involving vitamin E, selenium and protein. Inflammatory disease of the intestinal tract was also a major feature. Chronic colitis was particularly common in older marmosets. Pneumonia, otitis media, meningitis and brain abscesses were important pathological findings in home-bred marmosets and were commonly associated with bacterial infections, particularly Bordetella bronchiseptica and Klebsiella species. Trichospirura leptostoma within pancreatic ducts of wild-caught marmosets was the only significant parasitic disease encountered. Mycotic infections of the upper alimentary tract with Candida species were occasional findings in debilitated animals. No pathological features suggesting viral diseases were found.

Animals

An outbreak of Bordetella bronchiseptica pneumonia in a colony of common marmosets (Callithrix jacchus).

An outbreak of Bordetella bronchiseptica pneumonia occurred in a breeding colony of common marmosets (Callithrix jacchus). 16 animals, all except one under 12 months of age, died suddenly. Extensive lesions of pneumonia and pleurisy were found at necropsy and B. bronchiseptica was isolated from the nasopharynx, trachea and lungs. Older animals had only a mild rhinitis. Colonization of the nasal mucosa occurred in 71 of 156 marmosets.

Animals

A survey of the pathology of marmosets (Callithrix jacchus) under experiment.

A survey of the pathology of 567 laboratory-bred cotton-eared marmosets (Callithrix jacchus) is presented. There were few significant pathological changes in animals used in studies up to 6 months in duration, suggesting that the marmoset can be a useful non-human primate species for routine toxicology. The most common pathological changes encountered were chronic colitis, chronic thyroiditis and interstitial mononuclear infiltration in the kidney. No internal parasites were encountered, nor were any viral or bacterial diseases identified. Fungal disease was confined to a few cases of oesophageal mycoses. In a long term study a variety of pathological changes have been observed, including a 'wasting' syndrome, not related to skeletal muscle myopathy, with atrophy of the gastrointestinal tract, salivary glands and gonads, haemosiderosis and fatty change in the liver and osteoporosis. It is suggested that these changes may be related to protein deficiency and that the nutritional requirements of the marmoset require further investigation.

Animals

Systems for collection of urine in the captive common marmoset, Callithrix jacchus.

Two systems are described for the collection of 24 h urine samples from the common marmoset (Callithrix jacchus). Using 84 adult animals, 1210 24-h samples were collected. Mean urinary excretion was 14.4 +/- 7.5 ml/24 h (n = 1210, mean +/- SD). No differences were observed between sexes (for 52 females, 24 h volume = 15.1 +/- 8.0 ml; for 32 males, 24 h volume = 12.5 +/- 6.0 ml). No significant differences were observed between pregnant and non-pregnant females with respect to 24 h urine volume, and bilateral gonadectomy did not influence subsequent urinary excretion in either sex. For 161 pairs of observations, the intake of drinking water (11.7 +/- 10.2 ml/24 h) and the volume of urine excreted (12.6 +/- 7.1 ml/24 h) showed a positive correlation (r = 0.406 d.f. 159, P less than 0.001: y = 0.558x + 4.247).

Animals

Heinz bodies do not modify the membrane characteristics of common marmoset (Callithrix jacchus) erythrocytes.

The possibility was examined that the membrane function of erythrocytes obtained from healthy common marmosets (Callithrix jacchus) was modified by the presence in the cells of Heinz bodies. No significant differences were found in erythrocyte endogenous free malonyl dialdehyde (MDA) or reduced glutathione (GSH) between normal human erythrocytes and marmoset erythrocytes containing Heinz bodies. Membrane fluorescent chromolipids, surface charge and thiol levels were similar in both species but average membrane bulk lipid fluidity was slightly elevated in the marmosets. It was concluded that, in contrast to the situation in human erythrocytes, the presence of Heinz bodies in red cells of marmosets does not adversely affect the properties of the membrane.

Animals

Sex ratio and mortality in a laboratory colony of the common marmoset (Callithrix jacchus).

In a retrospective study sex ratio and mortality were analysed in a captive colony of common marmosets (Callithrix jacchus). Seven hundred and thirty-five infants in 294 litters (20 singletons, 119 twins, 140 triplets, 14 quadruplets) out of 57 breeding females were evaluated. The sex ratio at birth was 0.95 males:1.0 females. The frequency of males and females, as well as the sex composition of twins and triplets confirm the assumption of dizygotic twinning in the common marmoset. According to age at death, 9 categories were differentiated, with perinatal mortality being the highest. Once early infancy had passed the probability of a common marmoset infant of our colony reaching childhood is nearly 95%. Sixty per cent of all liveborn infants survived beyond 18 months. Mortality of infants at birth from primiparous mothers did not differ from that of pluriparous females, nor did the survival rate of infants with the filial generation the respective female had reached (F1 to F6). Females with a high ratio of triplets and quadruplets had a lower reproductive success than females with a majority of singleton or twin deliveries. Differential mortality between males and females was not observed. The frequency of stillbirths was not strongly related to parity, but was to litter-size. Most stillborn babies were seen in sets of quadruplets, most abortions in singletons. A normal socialization in a stable social environment, as well as not pairing the animals before they are fully adult, are considered important factors in good breeding success and infant survival.

Abortion, Veterinary

The facial artery of the common marmoset (Callithrix jacchus).

The facial artery and its ramifications in 7 adult common marmosets (Callithrix jacchus) were studied by the plastic injection method. The findings obtained are discussed in comparison with those for other primates. In the submandibular region, the facial artery arose from the external carotid artery at the height of the atlas via the linguofacial trunk on 10 of the total of 14 sides examined and independently on the other 4 sides. This common trunk always gave rise to the superior thyroid artery. The facial artery passed anterolaterally between the styloglossus muscle and the intermediate tendon of the digastricus muscle, giving off the styloglossal and the submandibular glandular branches, and anteroinferiorly medial to the pterygoideus medialis muscle. In a position anterior to this muscle, the submental artery and masseteric branch were derived. The submental artery gave off the medial pterygoid, the digastric, the cutaneous, the sublingual glandular and the mylohyoid branches, and then continued up to the median line, where it terminated to supply the genioglossus muscle. In the facial region, the facial artery passed anterosuperiorly along the anterior margin of the masseter muscle on 12 sides and away from it forwards on 2 sides, giving off the premasseteric branch in one of these 2 sides. It gave rise to the cutaneous, the buccal and the buccinator branches, the inferior labial artery and the communicating branch with the zygomatic artery. It terminated to divide into the superior labial and the naris lateral arteries, although the latter was lacking on 4 sides. The inferior labial artery gave off the mandibular marginal, the inferior labial marginal and the inferior labial glandular branches and terminated to anastomose with the mental artery. The superior labial artery divided into the superficial and deep branches, each of which continued as a nasal septal branch. The facial artery of the common marmoset usually ascended along the anterior margin of the masseter muscle and did not reach the medial angle of the eye.

Animals

Some pathways of xenobiotic metabolism in the adult and neonatal marmoset (Callithrix jacchus).

1. An oral dose of [carboxyl-14C]benzoic acid was excreted as hippuric acid or benzoylglucuronide by the adult marmoset Callithrix jacchus, and the relative proportions of the metabolites were dose-dependent over the range 1-100mg/kg body weight, the proportion of glucuronideincreasing with increasing dose level. 2. [14C]Aspirin was rapidly absorbed by the marmoset, giving peak blood levels within 1 h of dosing. Unchanged aspirin (trace to 5% dose) was excreted up to 3 h after dosage. Salicylic acid was the major metabolite, relatively little asalicyluric acid was excreted; glucuronic acid conjugates accounted for about 30% of the urinary metabolites and gentisic acid about 2%. 3. p-aminobenzoic acid administered to adult marmosets was excreted as p-acetamidobenzoic acid, the major urinary metabolite, unchanged p-aminobenzoates, p-aminobenzoyl glucuronide and p-aminohippuric acid each representing about 10% of the urinary metabolities; one-fifth of the p-aminohippurate was acetylated. 4. After oral administration of [35S]sulphadiazine to marmosets, peak blood levels of 35S occurred within 5 h dosing, and immediately fell to near-zero values at 24 h. Sulphadiazine and N-acetylsulphadiazine comprised 36% and 34% respectively of the urine 35S while the N1-glucuronide (tentative identification) accounted for 22%. Minor metabolites were the N4-glucuronide and N4-sulphate of sulphadiazine. 5. The extent of acetylation of sulphadimidine by marmosets corresponded to that of fast acetylators in man. 6. The metabolism of [14C]benzoate and of p-aminobenzoic acid in the neonatal marmoset was compared with that in similarly dosed neonatal rats.

Animals

Morphological and histochemical effects of 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD) on marmoset (Callithrix jacchus) testes.

The testes of marmosets (Callithrix jacchus), which had been treated with a single dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) (0.3 microgram to 10 micrograms/l kg body weight (BW)) were studied after 7 days using morphological and histochemical techniques. Light microscopic and electron microscopic examination revealed decreased intercellular contact in the germinal epithelium, as indicated first by enlarged intercellular spaces between the Sertoli's cells and between the Sertoli's cells and neighboring germ cells (i.e., spermatogonia and preleptotene spermatocytes), particularly in the basic compartment of the germinal epithelium. Second, decreased intercellular contact was indicated by the accumulation of premature spermatids and spermatocytes in the tubular lumen after TCDD treatment. The Sertoli's cells exhibited an increased amount of lipids, phagolysosomes, and vacuoles in their cytoplasm. Spermatids were frequently affected by TCDD, particularly during early spermiogenesis. These alterations included vacuolization of the cytoplasm and the development of additional germinal vesicles. This special effect on spermiogenesis became even more evident quantitatively by determination and counting of tubular stages in semithin sections. Tubular determination on the basis of the appearance of spermatids revealed that the ratio of tubular stages I to III became lower and that of stages V to VII became higher, dose dependently, indicating a maturation stop at the beginning of spermiogenesis caused by TCDD treatment. After TCDD treatment, Leydig's cells were morphologically unaffected, but histochemical investigations revealed decreased activity of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD). The sensitivity of the applied methods was different in view of the level of unaffection. The effect of Leydig's cells, as indicated by the decreased activity of 3 beta-HSD, had already been found at a dose of 1 microgram/kg BW TCDD, whereas clear-cut morphological and morphometrical effects were seen at 3 micrograms/kg BW for the first time. Moreover, with the special effect on spermiogenesis in marmoset monkeys, the findings demonstrate that the toxicity of TCDD on testicular morphology is species specific.

3-Hydroxysteroid Dehydrogenases

Characteristics of the light-induced phase response of circadian activity rhythms in common marmosets, Callithrix j. jacchus [primates-Cebidae].

Phase-response experiments using 1-h light pulses (LPs) of 1,100 lux applied under constant dim light of 0.3 lux were conducted with common marmosets, Callithrix j. jacchus, in order to obtain a complete phase-response curve established according to the common experimental procedure in a diurnal primate. Maximal phase delays of the free-running circadian activity rhythm (-90 min) were induced by LPs delivered at circadian time (CT) 12; e.g., during the beginning of the marmosets' rest time, maximal advances (+25 min) were elicited by pulses administered during the late subjective night at CT 21. In contrast to rodents, neither regular transient cycles nor regular period responses resulted from LP applications at different phases. To check whether the underlying period length affects the phase response in primates as well, the marmosets' circadian timing system was entrained to 25 h by a light:dark (LD) cycle of 12.5:12.5 h. The 1-h LPs were delivered during the first circadian cycle produced under constant dim light after the entraining LD periods. Here, LPs applied at CT 21 led to phase advances exceeding those measured during the steady-state free run. At CT 12, minor or no phase delays could be elicited. These findings show that the phase-shifting effect of LPs on the circadian system of marmosets is similar to that observed in other diurnal mammals. Some of the results indicate that in this diurnal primate, LP-induced phase shifts may be mediated in part by a light-induced increase in locomotor activity (arousal).

Animals

Junin virus infection of Callithrix jacchus: pathologic features.

Infection of Callithrix jacchus, a New World primate, with the prototype strain of Junin virus produced a severe disease. The animals developed multifocal hemorrhages and characteristic microscopic lesions such as meningoencephalitis, interstitial pneumonia, lymphocytic depletion of lymphatic tissue, hepatocytic necrosis, and a variable decrease in bone marrow cellularity. High virus concentrations correlated with lesions, and with the presence of viral antigenic determinants as revealed by immunofluorescent methods. With the exception of central nervous system damage, the morphological features and immunohistochemical and viral findings were similar to those recorded in human Argentine hemorrhagic fever.

Animals

Zonal differentiation of the marmoset (Callithrix jacchus) endometrium.

The differentiation of the marmoset (Callithrix jacchus) endometrium under different steroid hormone levels was investigated by electron microscopy and by the binding of different antibodies directed against collagen types. Based on differences in the glandular and interglandular compartments, the endometrium of sexually mature common marmosets consists of 3 zones: basal, adluminal and luminal. Hormone-dependent appearances are characterised. With low steroid concentrations, intercellular spaces between glandular epithelial cells occurred in the adluminal and the luminal areas. Epithelial cells of the basal region exhibited coated pits and phagolysosomes together with large apical protrusions. Under oestrogen dominance, phagolysosomes, fat vesicles and apical protrusions were evident in epithelial cells in the adluminal region. Secretory granules and concentric glycogen accumulations were a characteristic feature in epithelial cells of the adluminal and basal regions. With high progesterone concentrations, large empty vesicles were found with a higher frequency in adluminal than in basal epithelial cells. Using FITC-labelled antibodies against types V and VI collagen, binding was apparent adluminally in close vicinity to basement membranes, whereas reactivity was seen in the entire interglandular region of the basal area during this phase. Our findings indicate specific microenvironments with distinctive structural characteristics in the marmoset endometrium that are hormone dependent during all phases of the endometrial cycle. They are not related to menstruation, appear to be characteristic for primates and could reflect epithelial-mesenchymal interaction.

Animals

Age determination of common marmosets (Callithrix jacchus) by radiographic examination of skeletal development.

Seventy-four animals were examined radiographically to determine the skeletal development of the common marmoset (Callithrix jacchus) from 6 months of age. Twenty-one epiphyses were examined and five stages of ossification were described for each. The animals were divided into nine groups, according to age, and a table of the stage of ossification and age was produced, which may be used for determining the age of animals of unknown history.

Age Determination by Skeleton

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