The gut microbiome and metabolome of saddleback tamarins (Leontocebus weddelli): Insights into the foraging ecology of a small-bodied primate.

Body mass is a strong predictor of diet and nutritional requirements across a wide range of mammalian taxa. In the case of small-bodied primates, because of their limited gut volume, rapid food passage rate, and high metabolic rate, they are hypothesized to maintain high digestive efficiency by exploiting foods rich in protein, fats, and readily available energy. However, our understanding of the dietary requirements of wild primates is limited because little is known concerning the contributions of their gut microbiome to the breakdown and assimilation of macronutrients and energy. To study how the gut microbiome contributes to the feeding ecology of a small-bodied primate, we analyzed the fecal microbiome composition and metabolome of 22 wild saddleback tamarins (adult body mass 360-390 g) in Northern Bolivia. Samples were analyzed using high-throughput Illumina sequencing of the 16 S rRNA gene V3-V5 regions, coupled with GC-MS metabolomic profiling. Our analysis revealed that the distal microbiome of Leontocebus weddelli is largely dominated by two main bacterial genera: Xylanibacter and Hallella (34.7 ± 14.7 and 22.6 ± 12.4%, respectively). A predictive analysis of functions likely carried out by bacteria in the tamarin gut demonstrated the dominance of membrane transport systems and carbohydrate metabolism as the predominant metabolic pathways. Moreover, given a fecal metabolome composed mainly of glucose, fructose, and lactic acid (21.7 ± 15.9%, 16.5 ± 10.7%, and 6.8 ± 5.5%, respectively), the processing of highly fermentable carbohydrates appears to play a central role in the nutritional ecology of these small-bodied primates. Finally, the results also show a potential influence of environmentally-derived bacteria in colonizing the tamarin gut. These results indicate high energetic turnover in the distal gut of Weddell's saddleback tamarin, likely influenced by dominant bacterial taxa that facilitate dietary dependence on highly digestible carbohydrates present in nectar, plant exudates, and ripe fruits.

FILAMENTOUS FUNGI ISOLATED FROM THE FUR MICROBIOTA OF CALLITRICHIDS KEPT IN CAPTIVITY IN BRAZIL.

This study aimed to isolate filamentous fungi from the fur of primates of the genus Callithrix kept in the Centre for Rehabilitation of Wild Animals (CRWA) at the Tietê Ecological Park, São Paulo, SP, Brazil. Samples of the fur of 19 specimens of black-tufted marmosets (Callithrix penicillata) and 6 specimens of white-tufted-ear marmosets (Callithrix jacchus) were obtained by the square carpet technique. The samples were plated on Mycosel™ agar medium (Difco™) and incubated at 25°C for 21 days. The identification of each isolated mold was based on its macroscopic and microscopic features and followed classical recommendations. The following filamentous fungi were isolated: Penicillium spp. (76%), Cladosporium spp. (60%), Acremonium spp. (44%), Scopulariopsis spp. (24%), Aspergillus spp. (16%), Chrysosporium spp. (16%), and Fusarium spp. (8%). Dermatophyte fungi were not detected. We conclude that C. penicillata and C. jacchus kept in captivity are sources of potentially pathogenic filamentous fungi that may represent a risk factor for immunocompromised individuals who may eventually establish contact with them.

Isolation and identification of fungi from vaginal flora in three species of captive Leontopithecus.

The ability to reproduce in captivity is an essential component of lion tamarin (Leontopithecus) conservation programs. However, infections such as vaginitis, cervicitis, and endometritis are important diseases that may influence the reproduction of these animals. Therefore, it is important to detect continuous or occasional vaginal microbial populations, and to understand their potential role as an endogenous source of infection [Collins, 1964; Blue, 1983; Pugh et al., 1986]. Vaginal swabs were collected from 25 female tamarins of the three currently available species (L. rosalia, L. chrysopygus, and L. chrysomelas) at the Center of Primatology in Rio de Janeiro, Brazil. The swabs were processed according to standard mycological protocols, and isolates were biochemically characterized. Fungal isolates were recovered from 16 animals (64.0%). The results showed that 70.6% of the isolated microorganisms consisted of yeast, including three species of Candida (mainly C. glabrata). We suggest that this species is a resident member of the normal vaginal flora in Leontopithecus. Filamentous fungi (mainly from Trichosporon, Aspergillus, and Penicilliumgenera) constituted 29.4% of the isolates, and were considered to be transitory contaminants of the genital area. We suggest that colonization of the vaginal environment is related to the endocrine pattern associated with the reproductive status of these animals, but not to parity.

Identification, using sera from exposed animals, of putative viral antigens in livers of primates with callitrichid hepatitis.

Callitrichid hepatitis (CH) is an acute, frequently fatal viral hepatitis which affects members of the primate family Callitrichidae (R. J. Montali, E. C. Ramsay, C. B. Stephensen, M. Worley, J. A. Davis, and K. V. Holmes, J. Infect. Dis. 160:759-765, 1989; E. C. Ramsay, R. J. Montali, M. Worley, C. B. Stephensen, and K. V. Holmes, J. Zoo Wildlife Med. 20:178-183, 1989). Outbreaks of the disease occur in zoos and animal parks. In this study, CH-specific antigens were identified in the livers of infected animals by using immune sera from primates with CH and CH-exposed asymptomatic animals. Three CH-specific antigens with apparent molecular masses of 34, 54, and 65 kDa were identified. A polyclonal antiserum was raised against the 54-kDa antigen. These antigens were not found in the livers of uninfected animals and may be viral proteins. Our results suggest that at least five of the six outbreaks of CH considered here were caused by the same virus or by an antigenically related virus.

[Primates in the study of hepatitis viruses]. / Les primates dans l'étude des virus des hépatites.

There is no a conventional tissue culture system for the propagation of the hepatitis viruses and only some of them can be maintain in continuous cell culture in particular conditions. A transmissibility of hepatitis is limited to primates. The narrow host specificity may help to establish the classification of the hepatitis viruses and their mode of transmission. Moreover, the primate animal model provided the most accessible source of viruses and for clinical reasons presents the only opportunity for the studies of pathogenic mechanisms involving cellular immunity with allogenic restriction. The marmosets and chimpanzees susceptible to the hepatitis A and B viruses, respectively are the primates of choice for the experimental models. For the studies on parenterally transmitted NANB hepatitis the chimpanzee and some rhesus monkeys may provide an animal system. At last, most of the primates seem to be susceptible to agent responsible for the water-borne non A non B hepatitis.

Isolation and identification of strains of Bacteroides fragilis group from the digestive tract of Callithrix penicillata marmosets.

Thirty-five strains of the Bacteroides fragilis group were isolated from oral and intestinal samples from 5 wild caught, captive Callithrix penicillata. Nine oral strains of Bacteroides fragilis (7) and Bacteroides distasonis (2), and 26 intestinal strains of Bacteroides fragilis (14) and Bacteroides distasonis (12) were identified.

A new transmissible viral hepatitis of marmosets and tamarins.

Callitrichid hepatitis (CH) is a newly recognized, acute, fatal, epizootic disease of New World primates in the family Callitrichidae. Since 1980, 12 outbreaks of CH have occurred in US zoos, involving several callitrichid species including the endangered golden lion tamarin (Leontopithecus rosalia). CH was experimentally transmitted to common marmosets via a bacteria-free filtrate of liver from a naturally infected tamarin. All three inoculated marmosets developed an acute fatal disease with the characteristic clinical and histopathologic findings of CH. Human hepatotropic viruses that can infect the livers of callitrichids were not detected serologically in any of the experimentally infected marmosets. Enveloped viruslike particles 85-105 nm in diameter were observed in the rough endoplasmic reticulum and Golgi complex of hepatocytes from both naturally infected and experimentally inoculated animals. An immunoblot assay was developed using sera from tamarins exposed to natural outbreaks of CH and liver extracts from experimentally infected or control marmosets. A new CH-specific antigen was detected in the livers of naturally infected and experimentally inoculated marmosets but not controls. These results suggest that the etiologic agent of callitrichid hepatitis is a new primate hepatitis virus.

Epstein-Barr virus gene expression in malignant lymphomas induced by experimental virus infection of cottontop tamarins.

Inoculation of cottontop tamarins with a large dose of Epstein-Barr virus (EBV) leads to the induction of multiple EBV genome-positive lymphomas. These tumors have been characterized as oligoclonal or monoclonal large-cell malignant lymphomas that closely resemble the EBV genome-positive B-cell lymphomas that arise in human allograft recipients. The expression of latent and lytic EBV-encoded proteins was investigated in these virus-induced tamarin lymphomas and in derived cell lines. The tamarin tumors were found to express EBV nuclear antigen 1 (EBNA 1), EBNA 2, EBNA leader protein, and the latent membrane protein (LMP) as determined both by immunohistochemical staining and by immunoblotting. However, within the limits of the immunoblotting assays, no expression of the EBNA 3a protein family could be detected. Assays for lytic-cycle proteins by using both polyclonal human sera and monoclonal antibodies against viral capsid antigen, early antigen, and membrane antigen (gp340/220) showed minimal, if any, expression of these antigens in the lymphoma biopsies. In contrast, the cell lines derived from these lymphomas, even in early passage, expressed abundant levels of the lytic-cycle antigens and also expressed the EBNA 3a protein as well as EBNA 1, EBNA 2, EBNA leader protein, and LMP. This finding suggests that the virus-lymphoma cell interaction, in particular the switch to lytic cycle, is subject to some form of host control in vivo. The expression of EBNA 2 and LMP in these tamarin lymphomas strengthens their resemblance to posttransplant lymphomas in humans, since these human tumors are also EBNA 2 and LMP positive (L. S. Young, C. Alfieri, K. Hennessy, H. Evans, C. O'Hara, K. Anderson, A. Rickinson, E. Kieff, and J. I. Cohen, submitted for publication). Since both proteins are known to be important effector molecules of virus-induced B-cell growth transformation in vitro, their expression in these lymphomas constitutes the best evidence for a direct oncogenic role for EBV in vivo.

Studies of GB hepatitis agent in tamarins.

Three tamarins (Saguinus labiatus), two of which had previously been infected with hepatitis A virus and parenteral non-A, non-B hepatitis, were inoculated intravenously with the agent of GB hepatitis. All three animals developed alanine aminotransferase abnormalities 2 weeks after inoculation. Peak alanine aminotransferase levels were recorded 4 weeks postinoculation. These declined thereafter but continued to fluctuate at abnormal levels for 32 weeks. Liver biopsies showed liver cell swelling and inflammation with focal necrosis. Portal tracts and areas around central veins were heavily infiltrated with mononuclear cells. A fourth animal (no previous exposure to hepatitis viruses) inoculated with GB was killed on Day 15 postinoculation. Serum and extracts of liver and feces from this day were used as inocula for three other animals. Only the serum and liver extract transmitted GB hepatitis. The fecal specimen did not transmit and a fecal extract taken at a later date from another animal was also noninfectious. GB hepatitis virus is distinct from the viruses causing Type A and blood-borne non-A, non-B-hepatitis. Although the virus is present in serum and has previously been transmitted per os, it is not shed in feces.

Inoculation of New World primates with the human immunodeficiency virus.

The susceptibility of common marmosets and cotton-top tamarins to infection by HIV-2 in vivo was tested. One year and 19 months, respectively, post-inoculation, sera taken from three of four animals from each species are reactive for HIV-2 antibodies and HIV-specific nucleotide sequences were demonstrated in short-term cultures of PBL from two cotton-top tamarins. The animals remain in good health.

Rapid diagnosis and management of parainfluenza I virus infection in common marmosets (Callithrix jacchus).

During 1983 a severe episode of respiratory infection occurred in a marmoset colony at these laboratories. Of 91 marmosets, 69 showed clinical signs of disease, one died and nine were so ill that euthanasia was necessary. Eight were examined post mortem and all showed consolidation of the lungs. Laboratory studies were carried out in an attempt to establish the cause of the outbreak and an interstitial pneumonia was found in seven animals which were examined histologically. Direct electron microscopy of nasal swabs and lung samples revealed the presence of a high titre of a paramyxovirus, and subsequent immunofluorescence studies established that the particular paramyxovirus involved was parainfluenza virus type I. Subsequent studies showed that surviving affected animals had seroconverted to parainfluenza I virus while animals that had not been implicated in the outbreak had not.

Oral infection of a common marmoset with human T-cell leukemia virus type-I (HTLV-I) by inoculating fresh human milk of HTLV-I carrier mothers.

To obtain definitive evidence that milk-borne infection plays a critical role in the endemy or mother-to-child transmission of human T-cell leukemia virus type-I (HTLV-I), we inoculated concentrated fresh human milk cells obtained from HTLV-I carrier mothers into the oral cavity of a common marmoset (Callithrix jacchus). Twenty-eight milk samples were collected (5-10 ml each) from 17 carrier mothers in the first week after delivery. Cells in the milk were centrifuged down and resuspended in 1/10 vol of the milk fluid. The concentrated cell suspensions were successively inoculated into the oral cavity of a common marmoset. The marmoset was found to be seroconverted by indirect immunofluorescence assay at 2.5 months after the first inoculation of the milk (3.5 X 10(8) cells in total), and was later confirmed to be infected with HTLV-I by the detection of viral antigen expression in short-term cultures of its peripheral blood T-lymphocytes. The results strongly support the working hypothesis that milk-borne infection plays a significant role in the mother-to-child transmission of HTLV-I.

Attenuated Junin virus infection in Callithrix jacchus.

Twenty marmosets, male Callithrix jacchus, were used during this study. Fifteen of the marmosets were inoculated with 5,000 TCID50 of the attenuated XJC13 strain of Junin virus by intramuscular route and five were left as uninoculated controls. Animals were observed for a 420-day period. In order to carry out virologic, hematologic, serologic, and histologic studies the animals were bled and/or killed at different days post infection(pi). Results obtained showed that the attenuated strain produced an infection with no mortality or signs of illness. There was only a slight loss of weight at 18-40 days pi, which was soon recovered. Viremia was present from day 6 to 22, titers peaking at 4.0 log. Viral spread was limited to the lungs, spleen, lymph nodes, and bone marrow in the animal killed on day 14. No virus was found in the organs of the animal killed on day 23, and neither hematologic alterations nor pathologic lesions were seen in these monkeys except for ganglionar hypertrophy with immunoblast proliferation. Antigen was detected by immunofluorescence (IF) in lymph nodes, spleen, adrenals, lungs and brain. Neutralizing antibodies were detected from the third week onward. Protection conferred by the XJC13 strain proved effective when XJC13-inoculated monkeys were challenged with 1,000 TCID50 of the pathogenic XJ strain at days 60 or 380 pi, while normal controls died. When viral persistence was searched for on days 370, 390, and 420 pi, no infectious virus was detected, but viral antigen was seen in certain organs, which, however, lacked tissue damage.(ABSTRACT TRUNCATED AT 250 WORDS)

The guinea pig model for Argentine hemorrhagic fever.

Guinea pigs infected by the peripheral route with the XJ pathogenic strain of Junin virus showed viscerotropism mainly in reticulo-phagocytic rich organs. By immunofluorescence, heavy infection of reticular-phagocytic cells was demonstrated, supporting the leading role of these cell types. Absence of neurotropism was demonstrated by the inability to recover infectious virus, as well as the absence of antigens, immunoglobulins, or 3rd component of complement deposits, in cells, vessels, or meninges. The correlation between infectivity and antigen expression observed in organs, and the absence of evidence of immunopathologic mechanisms, strongly suggest a direct viral effect in these experimental conditions. The results show that infection of guinea pigs by the peripheral route is an adequate model for human Argentine hemorrhagic fever with the exception of central nervous system involvement. Comparisons are made with infections produced in guinea pigs by attenuated strains, as well as with the disease in primates and humans.

Monoclonal antibodies to hepatitis A virus.

This paper describes the development of monoclonal antibodies generated against hepatitis A virus (HAV). Monoclonal antibodies (MCABs) from two murine hybridoma cell lines were found to bind to an epitope recognized in the sera of patients recovering from infection with HAV. Ascites fluids containing MCABs from one hybridoma (H1 C19) inhibited a maximum of 70% of the 125I-labeled polyclonal human anti-HAV from binding to HAV-antigen in a competitive radioimmunoassay, indicating that the MCAB recognizes a major epitope of HAV. Monoclonal anti-HAV that was coated onto polystyrene beads was as effective as polyclonal antibodies in capturing HAV antigen from extracts of human feces, marmoset liver, and cultured cells. Radiolabeled MCAB was used to screen sera for anti-HAV. A collection of 117 sera was tested for total anti-HAV by competitive radioimmunoassay utilizing either 125I-labeled human polyclonal or mouse monoclonal antibody. Thirty-four specimens were similarly reactive by both systems, while the remainder were negative. Likewise, 28 specimens were similarly positive for IgM anti-HAV, and 12 specimens were negative using each of the two labeled antibodies. The data show that anti-HAV induced during disease is directed against a common major epitope of HAV and that MC anti-HAV can be used effectively as a reagent to detect these antibodies.

Immunogenicity of wild and attenuated varicella-zoster virus strains in pygmy marmosets.

Pygmy marmosets were inoculated with the low-passage parental strain or with the attenuated variant of OKA strain of varicella-zoster virus. No clinical signs were observed following inoculation and virus could not be isolated from tissues taken at several times after inoculation. A low-level antibody response developed in all animals. Three months after the first inoculation, all animals were challenged with the low-passage parental strain of virus. Animals primed originally with the parental strain developed higher booster responses than animals primed with the attenuated strain of virus. The results suggest that the parental and attenuated strains of varicella-zoster virus differ in their immunogenicity in pygmy marmosets.

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.