Occurrence and molecular characterization of Bartonella spp. and hemoplasmas in neotropical primates from Brazilian Amazon.

Little is known about the prevalence and genetic diversity of Bartonella spp. and hemoplasmas in nonhuman primates (NHP). The present study aimed to investigate the occurrence of and assess the phylogenetic position of Bartonella spp. and hemoplasma species infecting neotropical NHP from Brazilian Amazon. From 2009 to 2013, a total of 98 blood samples from NHP belonging to the Family Cebidae were collected in the island of São Luís, state of Maranhão, of which 87 NHP were from Wild Animal Screening Center (CETAS) in the municipality of São Luís, and 11 (9 Sapajus sp. and 2 Saimiri sciureus) were from NHP caught in the Sítio Aguahy Private Reserve. DNA samples were screened by previously described PCR protocols for amplifying Bartonella spp. and Mycoplasma spp. based on nuoG, gltA and 16S rRNA genes, respectively. Purified amplicons were submitted to sequencing and phylogenetic analysis. Bacteremia with one or more Bartonella spp. was not detected in NHP. Conversely, 35 NHP were PCR positive to Mycoplasma spp. The Blastn analysis of seven positive randomly selected sequencing products showed percentage of identity ranging from 95% to 99% with other primates hemoplasmas. The Maximum Likelihood phylogenetic analysis based on a 1510 bp of 16S rRNA gene showed the presence of two distinct clusters, positioned within Mycoplasma haemofelis and Mycoplasma suis groups. The phylogenetic assessment suggests the presence of a novel hemoplasma species in NHP from the Brazilian Amazon.

Absence of mutations associated with sulfa resistance in Pneumocystis carinii dihydropteroate synthase gene from non-human primates.

The dihydropteroate synthase (DHPS) gene from Pneumocystis carinii isolated from non-human primates was amplified using a polymerase chain reaction (PCR) and sequenced to analyse point mutations associated with sulfa resistance. P. carinii DHPS gene amplification was obtained from eight lung samples from five New World primate species and one Old World primate species. None of the animals had been exposed to sulfa drugs and only the wild-type P. carinii DHPS sequence at codons 55 and 57 was observed. These data support the hypothesis that high rates of DHPS mutants in P. carinii f. sp. hominis have arisen with increased use of sulfa drugs for P. carinii pneumonia prophylaxis.

Isolation and characterization of simian T-cell leukemia virus type II from New World monkeys.

Since the description of human T-cell leukemia virus type I (HTLV-I) and its simian counterpart, simian T-cell leukemia virus type I (STLV-I), the possible existence of other related simian retroviruses has been raised. Here, we report a new retrovirus, STLV-II, which we have identified in spider monkeys (Ateles fusciceps), a New World primate species. Initially, a recombinant HTLV-II envelope protein (RP-IIB) was used to identify anti-STLV-II antibodies in New World monkeys by Western blot (immunoblot) assays. Subsequently, the virus was characterized by Southern blot hybridization, which showed that STLV-II and HTLV-II have a high degree of nucleotide sequence homology but have different restriction enzyme patterns. Nucleotide sequence analysis of the pX-II region of STLV-II provirus revealed 3% variation with the corresponding region of HTLV-II. Electron micrographic studies revealed HTLV-like, type C retrovirus particles outside the cell membranes of STLV-II-infected cells. This study describes the first link between HTLV-II and a simian reservoir in the New World. Further molecular studies of STLV-II infection in different species of New World monkeys, especially from the wild, may provide valuable information about the origin and intragroup relationships of South American monkeys. Spider monkeys infected with STLV-II may serve as an important animal model for HTLV-II infection in humans.

Failure to detect human T-lymphotropic virus antibody in wild-caught New World primates.

We conducted a study to look for a simian counterpart of human T-lymphotropic virus (HTLV) in wild-caught monkeys in the Republic of Panama. Serum specimens were obtained from 102 monkeys (Ateles fusciceps, n = 75; Alouatta villosa, n = 18; and Cebus capucinus, n = 9) captured in Panama's Darien rain forest in 1979-1980. Specimens were screened for HTLV antibody by enzyme-linked immunosorbent assay, and reactive specimens were further tested by Western blot. None of the 102 specimens were seropositive for HTLV. Our findings provide no evidence for an HTLV-like virus in New World primates from Panama, but the sample size was small, and further studies are warranted.

Sequence analysis of herpes simplex virus gB gene homologs of two platyrrhine monkey alpha-herpesviruses.

Homologs of the herpes simplex virus gB gene were identified in two alpha-herpesviruses of platyrrhine monkeys, Herpesvirus saimiri 1 (HVS 1) and H. ateles 1 (HVA 1). These genes were cloned and sequenced in their entirety. Analysis of the predicted amino acid sequences indicated that the gB glycoproteins of these two viruses are of similar size, have 10 Cys residues and 5 potential N-linked glycosylation sites which align exactly with those in other primate alpha-herpesvirus gB polypeptides, and have a similar distribution of predicted secondary structural features, all of which indicate a conserved structure of the gB polypeptide. Alignment of these two gB sequences with those of four other primate alpha-herpesviruses (SA 8, B virus, HSV 1 and HSV 2) revealed localized regions of extensive sequence divergence as well as highly conserved regions. On comparison of the six primate virus gB sequences, the gBs of the two platyrrhine monkey viruses form a subgroup separate from that of the four catarrhine virus gBs. The degree of relatedness of the HVA 1 and HVS 1 gB sequences to each other was equivalent to the degree of relatedness between the human and the cercopithecine monkey virus gB sequences.

In vitro characteristics of Yersinia pseudotuberculosis of nonhuman primate origin.

Six strains of serotypes 1 or 2 of Y. pseudotuberculosis were isolated from dead squirrel monkeys, a cotton-top tamarin and a marmoset hybrid. All strains harboured a 71.6 kb plasmid, all were totally oxacillin-resistant and partially resistant to cephalosporins. Biochemically, serotypes 1 and 2 differed from each other in their beta-galactosidase production in a nonfermenter system, whereas the lack of rhamnose, maltose, salicin and trehalose fermentation seemed to be attributable to technical causes.

[Epidemiology of Klebsiella pneumoniae infections in 2 colonies of squirrel monkeys and lemurs]. / Epidémiologie des infections a Klebsiella pneumoniae dans deux élevages de singes-écureuils et de lémuriens.

After a short summary of the occurrence, sources, phenotypic characteristics, epidemiologic markers, virulence factors and pathogenicity of the 7 species of Klebsiella, the author reported (1) an infection due to K. pneumoniae K5 in a breed of squirrel monkey. These animals were suffering from sub-cutaneous abcesses (Pasteur Institute of Cayenne, French Guyana). (2) The second example refers a fatal infection due to K. pneumoniae K2 in a colony of lemurs, at Mulhouse zoological garden (East of France). Both animal colonies were protected against infection by the use of specific anti-K. pneumoniae capsular polysaccharide vaccine.

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.

Natural occurrence of black-pigmented Bacteroides species in the gingival crevice of the squirrel monkey.

The objective of this study was to determine whether the squirrel monkey (Saimiri scuireus) is indigenously colonized with black-pigmented bacteroides (BPB) resembling human Bacteroides gingivalis and Bacteroides intermedius (suspected periodontal pathogens) and to determine the usefulness of the squirrel monkey as an in vivo model for studying colonization by putative pathogens. We assayed the subgingival plaques of 138 monkeys of various ages and in four different colonies for the presence of anaerobic BPB microorganisms. We also tested half the animals for the presence of Actinobacillus actinomycetemcomitans. Clinical indices and levels of serum antibody to B. gingivalis were recorded. We detected BPB in 50% of the animals and A. actinomycetemcomitans in 69% of the animals. The presence of BPB was generally associated with increased age, increased gingival index, presence of calculus, and increased levels of serum antibody. These data indicate that the squirrel monkey may be a good model for studying the parameters of natural infection of the gingival crevice with suspected periodontopathogenic BPB microorganisms.

Attenuation of bovine parainfluenza virus type 3 in nonhuman primates and its ability to confer immunity to human parainfluenza virus type 3 challenge.

Bovine parainfluenza virus type 3 (PIV-3) was evaluated as a candidate live-virus vaccine to protect against infection with human PIV-3. The level of replication of bovine and human PIV-3 and the efficacy of immunization with bovine PIV-3 in protecting against subsequent challenge with human PIV-3 was evaluated in nonhuman primates. The duration and magnitude of replication of human and bovine PIV-3 in the upper and lower respiratory tracts of New World monkeys was similar, and animals infected with bovine PIV-3 developed resistance to challenge with human PIV-3. The replication of two bovine strains of PIV-3 was restricted 100- to 1000-fold in Old World primates but was sufficient to induce high levels of neutralizing antibody to human PIV-3. The combined properties of restricted replication and induction of a protective immune response to human PIV-3 in nonhuman primates make bovine PIV-3 a promising candidate for a live-virus vaccine to protect humans against disease caused by PIV-3.

Characterization of four herpesviruses isolated from owl monkeys and their comparison with Herpesvirus saimiri type 1 (Herpesvirus tamarinus) and herpes simplex virus type 1.

Four herpesviruses were previously isolated from four outbreaks of lethal disease in owl monkeys. All four isolates have been shown to be antigenically closely related to each other and to Herpesvirus saimiri 1 (HVS-1) by kinetic neutralizations. The owl monkey strains also share similarities to HVS-1 and to each other with respect to host range, growth cycles and molecular weights of peptides and of DNA fragments generated by restriction endonuclease (R.E.) digestion. R.E. analysis, however, can differentiate strains by the use of certain enzymes. All four isolates share a common G-C ratio percentage with HVS-1 of 67 per cent. On the basis of these findings, we believe that these owl monkey virus isolates are strains of HVS-1.

Nucleotide sequence analysis of squirrel monkey retrovirus reveals a novel primer-binding site for tRNALys1,2.

Nucleotide sequences of a DNA fragment containing the long terminal repeat (LTR) of squirrel monkey retrovirus (SMRV) were determined. Sequence analysis showed that the SMRV LTR is 456 base pairs (bp) long and is bounded by 2-bp inverted repeats. Within the U3 region, there are two 43-bp repeats and two 42-bp repeats which are homologous to each other. These repeats are likely to provide enhancer activities commonly observed in other enhancer sequences. Following the repeats are transcriptional regulatory sequences including a CAT box, a Goldberg-Hogness box, and a polyadenylation signal, all positioned within the U3 region of SMRV LTR. A 22-nucleotide sequence immediately downstream from the LTR was found to be complementary to tRNALys1,2, suggesting that tRNALys1,2 serves as the primer for the reverse transcription of SMRV viral RNA.

Comparison of the primate alphaherpesviruses. I. Characterization of two herpesviruses from spider monkeys and squirrel monkeys and viral polypeptides synthesized in infected cells.

Biological and biochemical properties of two neurotropic herpesviruses of New World monkeys--Herpesvirus saimiri type 1 (HVS-1) and Herpesvirus ateles type 1 (HVA-1)--were examined and compared. HVS-1 and HVA-1 both exhibited a time course of replication similar to another primate herpesvirus, SA 8. Both viruses grew rapidly and high titers of infectious virus were readily produced. HVS-1 and HVA-1 were also able to replicate efficiently in cell lines derived from a number of primate and non-primate species. Analysis of proteins synthesized in infected cells revealed the presence of over 30 virus-specific proteins ranging from less than 30,000 to over 200,000 daltons apparent molecular weight. Both viruses specified synthesis of a major capsid polypeptide of 148,000 daltons. Pulse labeling of cells during infection demonstrated temporal differences in the kinetics of synthesis of individual viral proteins and post-translational modification of a number of viral polypeptides. Glycosylated polypeptides synthesized in HVS-1 and HVA-1 infected cells were identified which ranged from approximately 49,000 to 120,000 daltons. Structural polypeptides of HVA-1 and HVS-1 virions were identified by SDS-PAGE analysis of purified virions. Taken together with clinical data on the diseases caused by these viruses, these studies indicate that HVS-1 and HVA-1 appear similar in many respects to both the human herpes simplex viruses and alphaherpesviruses of other primates.

Calicivirus isolation from three species of primates: an incidental finding.

Calicivirus isolations were made from 3 species of subhuman primates. Viruses were recovered from gingival lesions associated with periodontal disease in a spider monkey, from the oropharynx of a healthy silver leaf langur, and from the spleen of a lowland gorilla that had died of systemic coccidioidomycosis. Based on the results of cross-neutralization tests, all 3 isolates were serologically indistinguishable from a primate calicivirus Pan paniscus type 1. These isolations appeared to be incidental in nature and could not be associated causally with any specific disease entity.

Enterobactérias detectadas em primatas capturados na regiäo amazônica do Brasil / Enterobacteriaceae detected in primates captured in the Amazon Region of Brazil

A partir da implantaçäo do Centro Nacional de Primatas (CENP), em Belém, Pará, Brasil, proporcionou o estudo da microflora intestinal de 77 símios (38, C.apella; 14, S.sciureus; 20, C.a.argentata e 5, A.trivirgatus), através de coprocultivos realizados imediatamente após a captura desses animais na floresta amazônica brasileira e depois de 6 meses em cativeiro. Proteus mirabilis, Escherichia coli, Proteus morganii, Proteus vulgaris, Klebsiella pneumoniae, Proteus rettgeri, Enterobacter cloacae, Citrobacter freundii, Enterobacter aerogenes e Edwaedsiella tarda foram isolados dos primeiros coprocultivos procedidos logo após a captura dos símios. Salmonella worthington foi isolada de um exemplar de C.a.argentata em cativeiro

Hemobartonellosis in squirrel monkeys (Saimiri sciureus) in a domestic breeding colony: case report and preliminary study.

Infection with Hemobartonella sp was diagnosed in a colony-born squirrel monkey with normocytic, normochromic anemia and pronounced punctate erythrocytic basophilic stippling on Wright's-Giemsa stained blood films. The diagnosis was confirmed using transmission electron microscopy. Two randomly selected colony-born squirrel monkeys were splenectomized in an effort to activate and detect possible latent hemobartonellosis . One monkey became parasitemic 12 days following splenectomy. The second monkey was inoculated on day 14 with 1 ml of whole blood from an infected, but nonparasitemic monkey and developed overt parasitemia 3 days later (day 17 following splenectomy). Infections in the latter two monkeys were confirmed using scanning electron microscopy.

[Detection of antigenic determinants of the major structural proteins of various mammalian type C retroviruses in the leukocytes of patients with hemoblastoses and in healthy donors by the ELISA method]. / Opredelenie antigennykh determinant osnovnykh virusnykh belkov nekotorykh retrovirusov tipa C mlekopitaiushchikh v leikotsitakh bol'nykh gemoblastozami i zdorovykh liudei metodom ELISA.

Using the enzyme-labelled antibodies it was shown that extracts from leukocytes of patients with leukemias and healthy donors contain antigenic determinants related to major virus protein of mammalian C type oncornaviruses SSV/SSAV, BaEV, FeLV and RLV. The highest antigen activity in patients was detected in the feline leukemia (FeLV) system, and the least one--in the system of murine leukemia (RLV). Common antigenic determinants of major virus protein of the viruses under study were detected in all groups of patients with haemoblastoses and healthy donors, but the amount of positive results was considerably higher in the group of patients as compared to that in the group of donors.

[Lack of viral persistence in 2 Cebus sp]. / Falta de persistencia del virus en dos Cebus sp.

Two Cebus sp surviving acute Junin virus infection, one after intramuscular inoculation with pathogenic XJ and the other after intracerebral inoculation with XJ Clone 3, failed to exhibit persistent infection. Although treatment with immunosuppressive drugs was carried out, no Junin virus was detected in blood or organs in spite of blind passages in mice for the former as well as cocultivation with permissive Vero cells for the latter. Viremia was also ruled out by immunofluorescence on BHK/21 cell culture. These findings correlate with the lack of long-lasting virus in blood observed in humans following the acute phase of Argentine Hemorrhagic Fever.

Evaluation of the A/Seal/Mass/1/80 virus in squirrel monkeys.

An influenza A virus isolated from seals [A/Seal/Mass/1/80 (H7N7)] and an isolate of this virus obtained from a human conjunctiva were evaluated for replication and virulence in squirrel monkeys. When the seal virus was administered intratracheally, it replicated in lungs and nasopharynges and induced illness almost to the same extent that a human influenza A virus [A/Udorn/72 (H3N2)] did. In one monkey that died of pneumonia, the seal virus was recovered from spleen, liver, and muscle as well as lung. After conjunctival administration in monkeys, the seal virus replicated to a peak titer in the conjunctivae 30-fold greater than that attained by the human virus, but this difference was not statistically significant. In contrast, the seal virus replicated less well than the human virus in the tracheae and nasopharynges when administered by the conjunctival route. These results indicate that the seal virus can replicate efficiently in primates, that it can spread systemically, and that it might differ from human virus in being able to replicate slightly better in primate conjunctival tissue.

Experimental infection of the New World owl monkey (Aotus trivirgatus) with hepatitis A virus.

Epidemiological studies have demonstrated the susceptibility of the owl monkey (Aotus trivirgatus) to hepatitis A virus, but have not shown an association between infection and histopathological or chemical evidence of liver disease. Therefore, 12 seronegative, colony-bred monkeys were inoculated intravenously with a fecal suspension containing either PA33 strain hepatitis A virus (a strain recovered from a naturally infected Aotus sp.) or HM-175 virus (recovered from a human). Viral antigen was detected by radioimmunoassay in the feces of six monkeys 6 to 17 days after inoculation with PA33 virus, and by 9 to 21 days serum aminotransferase activities were significantly elevated in each. Antibody to the virus developed in each monkey by 28 days after inoculation. Similar findings were noted in five of six monkeys inoculated with HM-175 virus, although the incubation period preceding aminotransferase elevations was somewhat longer (25 to 39 days). Liver biopsies obtained from the 11 infected monkeys demonstrated mild to moderate portal inflammation, as well as random areas of focal necrosis and inflammation extending outward from the portal region. These data confirm the susceptibility of Aotus sp. to hepatitis A virus and indicate that the infection of this primate provides a useful animal model of human hepatitis A.