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A multidisciplinary panel of experts in March 2003 developed a risk assessment model to describe the likelihood of cross-species transmission of simian foamy virus (SFV) from macaques to visitors at a monkey temple in Bali, Indonesia. The model does predict that the risk of being infected with SFV among visitors to a monkey temple is not negligible, and it can be conceived as the first step in a process whereby non-human primate (NHP)-human cross-species disease transmission can be examined.

The taxonomy of herpesviruses has been updated by the International Committee on Taxonomy of Viruses (ICTV). The former family Herpesviridae has been split into three families, which have been incorporated into the new order Herpesvirales. The revised family Herpesviridae retains the mammal, bird and reptile viruses, the new family Alloherpesviridae incorporates the fish and frog viruses, and the new family Malacoherpesviridae contains a bivalve virus. Three new genera have been created in the family Herpesviridae, namely Proboscivirus in the subfamily Betaherpesvirinae and Macavirus and Percavirus in the subfamily Gammaherpesvirinae. These genera have been formed by the transfer of species from established genera and the erection of new species, and other new species have been added to some of the established genera. In addition, the names of some nonhuman primate virus species have been changed. The family Alloherpesviridae has been populated by transfer of the genus Ictalurivirus and addition of the new species Cyprinid herpesvirus 3. The family Malacoherpesviridae incorporates the new genus Ostreavirus containing the new species Ostreid herpesvirus 1. [ABSTRACT FROM AUTHOR]

Contact between humans and nonhuman primates (NHPs) frequently occurs at monkey temples (religious sites that have become associated with free-ranging populations of NHPs) in Asia, creating the potential for NHP–human disease transmission. In March 2003 a multidisciplinary panel of experts participated in a workshop designed to model the risk of NHP–human pathogen transmission. The panel developed a risk assessment model to describe the likelihood of cross-species transmission of simian foamy virus (SFV) from temple macaques (Macaca fascicularis) to visitors at monkey temples. SFV is an enzootic simian retrovirus that has been shown to be transmitted from NHPs to humans. In operationalizing the model field data, laboratory data and expert opinions were used to estimate the likelihood of SFV transmission within this context. This model sets the stage for a discussion about modeling as a risk assessment tool and the kinds of data that are required to accurately predict transmission. Am. J. Primatol. 68:934–948, 2006. © 2006 Wiley-Liss, Inc. [ABSTRACT FROM AUTHOR]

Monkey B virus (Macacine herpesvirus 1; BV) is endemic in macaques. BV (a BSL4 agent) is the primary zoonotic concern for persons working with macaques in research, and human BV infections frequently are fatal. We assessed the use of a BSL2 baboon herpesvirus (Papiine herpesvirus 1; HVP2) for predicting the drug sensitivity of BV by comparing the sensitivity of the 2 viruses to 12 antiherpetic drugs. Plaque reduction assays showed that 4 drugs (HBPG, BVdU, PFA, and BrdU) were ineffective against both viruses. Of the 8 effective drugs, both viruses were most sensitive to TFT, whereas sensitivity to the remaining 7 drugs varied between BV and HVP2 as well as between strains of HVP2. In addition, the efficacy of 5 drugs (ACV, PCV, GCV, CDV, and EDU) was tested by using a murine model. ACV and EDU were completely ineffective against both HVP2 and BV, and high doses of PCV only delayed death by a few days. GCV and CDV both protected mice against death, and CDV also prevented the development of neurologic symptoms. When the initiation of drug therapy was delayed until after virus gained access to the CNS, both GCV and CDV were ineffective. The similarity of the drug sensitivities of HVP2 and BV in both models validates the use of HVP2 as a BSL2 level model that can be used to predict drug sensitivity of BV. The greater efficacy of CDV relative to GCV suggests the potential for use of CDV in the treatment of zoonotic BV infections.

Two young female baboons naturally infected with simian T-lymphotropic virus type 1 (STLV1) were euthanized due to chronic respiratory disease that was unresponsive to treatment. Massive lymphocytic infiltration of the lung interstitium suggested a diagnosis of STLV-associated lymphoma. In each case, the diagnosis was confirmed through inverse PCR (IPCR) that detected monoclonally integrated STLV1 provirus in cellular DNA extracted from lymphoma tissue and peripheral blood cells (PBC). One dominant STLV1-infected T-cell clone and 3 minor clones were detected in PBC from each baboon. Using archived PBC DNA and primers within the proviral genome and chromosomal DNA flanking the STLV1 integration sites in PCR analyses, we determined that the dominant clone in one baboon had first appeared approximately 8 mo after infection and had circulated for 4 y before clinical disease developed. ELISA testing of archived serum revealed that both baboons seroconverted to the p19 and p24 gag proteins and the envelope gp46 protein but not to the viral tax protein. Titers to p24 and gp46 rose significantly after infection and remained relatively constant until death, whereas titers to p19 increased with time. Although spontaneous STLV1-associated lymphomas have been described in baboons, the STLV1-associated lymphomas described here occurred in 2 relatively young baboons, both of whom had become infected with STLV at 3 to 4 y of age and developed lymphoma within 5 y of infection.

We undertook establishing an SPF baboon colony in response to requests from researchers. To enable the widest possible future use of SPF baboons, our aim was to develop an SPF colony of baboons (Papio hamadryas anubis) free of 12 target viruses: 5 herpesviruses, 4 retroviruses, simian virus 40, measles, and monkeypox. Infant baboons were removed from their mothers within 24 h of birth and nursery-reared. Groups of 3 to 8 age-matched conspecifics were isolated in separate rooms for 1 y while undergoing repeated testing for target viruses. During the initial 7 y of the SPF program, 171 infants were enrolled, of which 76 (44.4%) subsequently were removed from the program. Of those removed, 54 (71.0%) were culled due to breaks in virus-free status, 12 (15.8%) died of various causes, 4 (5.3%) developed seizures, and 6 (7.9%) were removed for other reasons. The most problematic viruses were baboon cytomegalovirus (25.9% of culls), Herpesvirus papio 1 (51.9%), and simian foamy virus (7.4%). Using conspecific groups of 3 to 4 infants reduced first-year program losses as compared with groups of 6 to 8. There have been 17 births in the SPF colony, and all these infants have been free of all target viruses since birth. On the basis of these results, early removal of infants from their dams, housing in small peer groups, frequent virus testing, and aggressive culling of virus-positive animals is an effective approach for development of a baboon colony free of multiple viruses.

Macaques are a particularly valuable nonhuman primate model for a wide variety of biomedical research endeavors. B virus (Cercopithecine herpesvirus 1; BV) is an alpha-herpesvirus that naturally infects conventional populations of macaques. Serious disease due to BV is rare in macaques, but when transmitted to humans, BV has a propensity to invade the central nervous system and has a fatality rate greater than 70% if not treated promptly. The severe consequences of human BV infections led to the inclusion of BV in the original NIH list of target viruses for elimination by development of specific pathogen-free rhesus colonies. In macaques and especially in humans, diagnosis of BV infection is not straightforward. Furthermore, development and maintenance of true BV specific pathogen-free macaque colonies has proven dif cult. In this overview we review the natural history of BV in macaques, summarize what is known about the virus at the molecular level, and relate this information to problems associated with diagnosis of BV infections and development of BV-free macaque colonies.

Changes in levels of antibody to B virus (Cercopithecine herpesvirus 1; BV) were examined in BV-positive macaques by ELISA. We observed increases in anti-BV IgG titers in a BV-infected cynomolgus monkey after overseas transportation by air and in a rhesus monkey after transfer from an outdoor group cage to an indoor individual cage. Although shedding of infectious virus was not examined, the increase in antibody titer suggested reactivation of BV. Interestingly, we also found an increase in anti-BV IgG levels during the breeding season in male but not female Japanese macaques kept in an enclosed outdoor colony. Further studies should be performed to investigate whether reactivation of BV led to the observed increase in the anti-BV antibody titer.

We investigated the prevalence, distribution, and transmission of simian T-lymphotropic virus type 1 (STLV1) in a baboon breeding colony over a 4-y period. We used polymerase chain reaction amplification of the proviral tax gene to assess the infection status of 272 animals housed in 4 separate corrals. Sequencing the proviral envelope gene from individual baboons detected several molecular subtypes (genotypes) of STLV1. At the start of the study, 31% (54 of 176) of all baboons were infected; the majority of infections (91%) were in mature females, with only 3 of 12 mature males and 2 of 48 infants and juveniles being infected. Over the next 4 years, 41 new infections were diagnosed. Of these, 83% occurred in sexually mature female baboons (at least 3 y of age), 17% in infants and juveniles (younger than 3 y), and 0% in mature males. The 7 infections in juveniles were probably derived from mother-to-infant transmission because mother-infant pairs consistently were infected with the same viral genotype. Of the 34 new infections in sexually mature female baboons, the genotyping data showed that 25 (73%) originated from other infected females as opposed to males. Male-to-female sexual transmission may have accounted for the remaining 9 new infections. There was no evidence of female-to-male sexual transmission. The high percentage of female-to-female transmission of STLV1 in our baboons was unexpected; we speculate that transmission may have occurred due to blood contamination from biting during aggressive behavior between females in establishing hierarchical dominance.

In vivo studies with highly pathogenic viruses prompt concerns regarding the persistence of infectious virus in pathology specimens. Although formalin fixation of tissues may inactivate infectious virus, fixation may also degrade viral nucleic acid and antigens, thereby limiting detection of virus in tissues by polymerase chain reaction (PCR) amplification or immunohistochemistry (IHC). We sought to: 1) assess the rate of inactivation of infectious virus in tissue specimens during formalin fixation, 2) assess IHC recognition of viral antigens and PCR detection of viral DNA after long-term (14 d) formalin fixation, and 3) investigate microtome contamination by DNA carry-over to subsequently sectioned tissues. Infectious baboon herpesvirus HVP2 could be recovered from fresh tissues of infected mice but not those fixed in formalin for >/=24 h. The intensity of IHC staining of viral antigen was unaffected by the duration of formalin fixation. PCR detection of viral DNA was negatively impacted by formalin fixation and/or heat inherent to paraffin processing; however, amplification of very short DNA sequences using real-time PCR was not affected. Lastly, microtome contamination by viral DNA was demonstrated by PCR screening of uninoculated control tissues that were sectioned after sectioning infected tissues. In summary, infectious virus is inactivated after only 24 h of formalin fixation whereas IHC staining remains sensitive in tissues fixed for up to 14 d. Formalin fixation does degrade DNA, but viral DNA can be detected by PCR amplification of very short DNA sequences. In addition, viral DNA can contaminate a microtome knife such that subsequently sectioned uninoculated control tissues exhibit false positive PCR amplification.

Here we describe the unusual finding of herpesvirus pneumonia in a 7-d-old infant baboon (Papio hamadryas anubis). This animal had been separated from its dam the morning of its birth and was being hand-reared for inclusion in a specific pathogen-free colony. The baboon was presented for anorexia and depression of 2 d duration. Physical examination revealed a slightly decreased body temperature, lethargy, and dyspnea. The baboon was placed on a warm-water blanket and was given amoxicillin-clavulanate orally and fluids subcutaneously. The animal's clinical condition continued to deteriorate despite tube feeding, subcutaneous fluid administration, and antibiotic therapy, and it died 2 d later. Gross necropsy revealed a thin carcass and severe bilateral diffuse pulmonary consolidation. Histopathology of the lung revealed severe diffuse necrotizing pneumonia. Numerous epithelial and endothelial cells contained prominent intranuclear herpetic inclusion bodies. Virus isolated from lung tissue in cell culture was suspected to be Herpesvirus papio 2 (HVP2) in light of the viral cytopathic effect. Real-time polymerase chain reaction (PCR) analysis and DNA sequencing of PCR products both confirmed that the virus was HVP2. This case is interesting because the age at onset suggests perinatal transmission at or immediately after birth, and the disease course suggests inoculation of the virus into the respiratory tract.

Cercopithecine herpesvirus 16 (Herpesvirus papio 2; HVP2) is an alpha-herpesvirus of baboons (Papio spp.) that generally causes minimal to inapparent disease in the natural host species. HVP2 is very closely related genetically and antigenically to Cercopithecine herpesvirus 1 (monkey B virus; BV) of macaques, which is well known for its extreme lethality in nonmacaque species including humans. Preliminary evidence suggests that a mouse model of HVP2 would be an excellent tool for studying zoonotic BV infections. Although the pathogenicity of different BV isolates in mice spans the full range of severity from apathogenic to extremely neurovirulent, testing of multiple HVP2 isolates revealed only two distinct phenotypes in mice regardless of route of inoculation: apathogenic (HVP2ap) and highly neurovirulent (HVP2nv). For the HVP2nv mouse model to truly reflect BV infection in both its natural host and the differential pathogenicity of BV in aberrant host species, HVP2nv should not produce severe disease in its natural host. To test this, juvenile baboons were inoculated with doses of 10(6) or 10(4) plaque-forming units of HVP2ap or HVP2nv by using an oral subdermal inoculation route. Parameters followed included the appearance of lesions, shedding of infectious virus, general health, and the immune response to the infection. Regardless of the inoculum dose used, no differences were noted between the two HVP2 subtypes in baboons in any of the parameters measured. These findings further support the use of the HVP2nv mouse system as a model to elucidate and study the viral determinants associated with cross-species BV neurovirulence.

Serologic testing for antibody to monkey B virus (BV) in macaque sera is problematic due to the biohazardous nature of BV antigens. Herpesvirus papio 2 (HVP2), a herpesvirus of baboons, is nonpathogenic to humans and is genetically and antigenically more closely related to BV than is human herpes simplex virus 1. This paper describes the results of our in-house laboratory that compared a BV antigen-based enzyme-linked immunosorbent assay (ELISA) by commercial testing laboratory and an HVP2-based ELISA in our laboratory by using 447 sera from 290 rhesus monkeys. The HVP2-based ELISA identified as positive 99.11% of the sera identified as BV-positive by the BV ELISA. The BV antigen-based ELISA identified as positive 98.21% of the sera identified as BV-positive by the HVP2-based ELISA. The HVP2 ELISA also identified two BV-negative and six BV-equivocal sera as positive. Both ELISAs identified the same 85 negative and three equivocal samples as negative and equivocal, respectively. The high degree of correlation (weighted kappa coefficient, 0.94) between the two tests indicates that the HVP2 ELISA is a sensitive and reliable assay for in-house testing of the BV status of rhesus monkeys.

Little is known about the natural history of herpesviruses indigenous in baboons. Here, we describe the development of ELISAs for five herpesviruses. These assays were used to test more than 950 serum samples collected from approximately 210 infant/juvenile and 130 adult baboons in a captive breeding colony over a period of seven years. Results indicated that baboon cytomegalovirus, lymphocryptovirus, and rhadinovirus are transmitted efficiently within the colony and are acquired at an early age. Baboon alpha-herpesvirus HVP2 and polyomavirus simian virus 40 (SV40) were acquired later and by fewer juveniles than were the other three herpesviruses. More than 60% of baboons acquired HVP2 before reaching sexual maturity, indicating that oral infection of infants and juveniles, rather than sexual transmission between adults, is the predominant mode of transmission for this virus. Antibody to simian varicella virus (SVV) was found in about 40% of baboons. SVV was acquired principally by infants and juveniles; few adults seroconverted despite seronegative adults being in constant contact with infants and juveniles undergoing primary infection. Time of seroconversion was not statistically correlated to specific individual herpesviruses, suggesting that each virus is acquired as an independent infection event rather than multiple viruses being acquired at the same time. Several baboons that were delivered by cesarean section and were housed separate from, but in close proximity to, other baboons remained free of many or all viruses for several years, suggesting that, similar to human herpesviruses, baboon herpesviruses and SV40 are transmitted principally by direct contact.

Detection of infectious viruses in clinical samples typically relies on daily examination of inoculated cell cultures for appearance of virus-induced cytopathic effect (CPE), with subsequent immunologic or genetic analysis to identify the specific virus producing the CPE. Performing virus isolation on samples suspected of containing Cercopithecine herpesvirus 1 (monkey B virus [BV]) is dangerous due to the extreme neuropathogenicity of this virus in humans, and minimally requires biosafety level 3 (BSL-3) facilities. To provide a safer method of detecting infectious BV in clinical samples, the eucaryotic green fluorescent protein (GFP) coding sequence was flanked with BV sequences containing transcriptional control elements. This construct was placed into a stealth vector and transfected into Vero cells, then stable transformed cell lines were selected. These cells express GFP only when infected with BV or other related primate herpesviruses. Expression of GFP allows detection of infectious BV in cultures sooner and more reliably than does standard microscopic observation for CPE. The ability to detect BV by GFP expression eliminates the need for further testing to identify the virus as an alpha-herpesvirus following development of CPE, thus allowing cell cultures to be sealed at inoculation. Although not entirely specific for BV, this cell line will make detection of infectious BV in samples collected from macaques safer to perform.

By adding betaine to the PCR mixture, we previously established a PCR method to amplify a DNA segment of the glycoprotein G gene of B virus (BV) derived from a rhesus macaque. We have found that DNA of other BV strains derived from cynomolgus, pigtail, and lion-tailed macaques can also serve as the template in our PCR assay. Under the same conditions no product was obtained with DNA of simian agent 8 of green monkeys and Herpesvirus papio 2 of baboons, or the human herpes simplex viruses types 1 and 2. Thus, this PCR method is useful to discriminate BV from other closely related primate alphaherpesviruses.