Stress in cynomolgus monkeys (Macaca fascicularis) subjected to long-distance transport and simulated transport housing conditions.

The stress associated with transportation of non-human primates used in scientific research is an important but almost unexplored part of laboratory animal husbandry. The procedures and routines concerning transport are not only important for the animals' physical health but also for their mental health as well. The transport stress in cynomolgus monkeys (Macaca fascicularis) was studied in two experiments. In Experiment 1, 25 adult female cynomolgus monkeys were divided into five groups of five animals each that received different diets during the transport phase of the experiment. All animals were transported in conventional single animal transport cages with no visual or tactile contact with conspecifics. The animals were transported by lorry for 24 h at ambient temperatures ranging between 20 degrees C and 35 degrees C. Urine produced before, during and after transport was collected and analysed for cortisol by enzyme-linked immunosorbent assay (ELISA). All monkeys exhibited a significant increase in cortisol excretion per time unit during the transport and on the first day following transport.Although anecdotal reports concerning diet during transport, including the provision of fruits and/or a tranquiliser, was thought likely to influence stress responses, these were not corrobated by the present study. In Experiment 2, behavioural data were collected from 18 cynomolgus macaques before and after transfer from group cages to either single or pair housing, and also before and after a simulated transport, in which the animals were housed in transport cages. The single housed monkeys were confined to single transport cages and the pair housed monkeys were kept in their pairs in double size cages. Both pair housed and singly housed monkeys showed clear behavioural signs of stress soon after their transfer out of their group cages.However, stress-associated behaviours were more prevalent in singly housed animals than in pair housed animals, and these behaviours persisted for a longer time after the simulated transport housing event than in the pair housed monkeys. Our data confirm that the transport of cynomolgus monkeys is stressful and suggest that it would be beneficial for the cynomolgus monkeys to be housed and transported in compatible pairs from the time they leave their group cages at the source country breeding facility until they arrive at their final laboratory destination in the country of use.

Isolation and partial characterization of a lentivirus from talapoin monkeys (Myopithecus talapoin).

We have identified a novel lentivirus prevalent in talapoin monkeys (Myopithecus talapoin), extending previous observations of human immunodeficiency virus-1 cross-reactive antibodies in the serum of these monkeys. We obtained a virus isolate from one of three seropositive monkeys initially available to us. The virus was tentatively named simian immunodeficiency virus from talapoin monkeys (SIVtal). Despite the difficulty of isolating this virus, it was readily passed between monkeys in captivity through unknown routes of transmission. The virus could be propagated for short terms in peripheral blood mononuclear cells of talapoin monkeys but not in human peripheral blood mononuclear cells or human T cell lines. The propagated virus was used to infect a naive talapoin monkey, four rhesus macaques (M. mulatta), and two cynomolgus macaques (M. fascicularis). All animals seroconverted and virus could be reisolated during a short period after experimental infection. A survey of SIVtal-infected captive talapoin monkeys revealed a relative decrease in CD4(+) cell numbers in chronically (>2 years) infected animals. No other signs of immunodeficiency were observed in any of the infected animals. PCR amplification followed by DNA sequencing of two fragments of the polymerase gene revealed that SIVtal is different from the presently known lentiviruses and perhaps most related to the SIV from Sykes monkeys.

Simian T-lymphotropic virus type I infection among wild-caught Indonesian pig-tailed macaques (Macaca nemestrina).

Evidence for the presence of simian T-lymphotropic viruses (STLV-I) was identified in live-caught pig-tailed macaques from two locations in southern Sumatra, Indonesia. Of 60 animals tested, 13.3% of the animals showed seroreactivity to HTLV-I/II enzyme-linked immunosorbent assay (ELISA) antigens. Of these, 75% showed indeterminate reactivity and 25% showed positive reactivity to HTLV-I/II Western blot antigens. Polymerase chain reaction (PCR) analysis of 6 of 8 seroreactive monkeys' peripheral blood mononuclear cell (PBMC) DNA showed production of proper size molecular weight product that hybridized specifically to an STLV-I tax gene-specific probe. Phylogenic analyses of tax gene fragment sequences from the PCR products of two samples, 930287 and 930306, indicated that these animals were infected with retroviruses related to those of the Asian STLV-I clade.

The tax gene sequences form two divergent monophyletic lineages corresponding to types I and II of simian and human T-cell leukemia/lymphotropic viruses.

Evolutionary associations of human and simian T-cell leukemia/lymphotropic viruses I and II (HTLV-I/II and STLV-I/II) are inferred from phylogenetic analysis of tax gene sequences. Samples studied consisted of a geographically diverse assemblage of viral strains obtained from 10 human subjects and 20 individuals representing 12 species of nonhuman primates. Sequence analyses identified distinct substitutions, which distinguished between viral types I and II, irrespective of host species. Phylogenetic reconstruction of nucleotide sequences strongly supported two major evolutionary groups corresponding to viral types I and II. With the type I lineage, clusters were composed of strains from multiple host species. A genetically diverse, monophyletic lineage consisting of eight new viral strains from several species of Asian macaques was identified. The second lineage consisted of a monophyletic assemblage of HTLV-II/STLV-II strains from Africa and the New World, including an isolate from a pygmy chimp (Pan paniscus) as an early divergence within the lineage. High levels of genetic variation among strains from Asian STLV-I macaque suggest the virus arose in Asia. Evidence of the origin of the type II virus is less clear, but diversity among HTLV-II variants from a single isolated population of Mbati villagers is suggestive but not proof of an African origin.

Tinjil island, a natural habitat breeding facility of simian retrovirus-free Macaca fascicularis.

Institut Pertanian Bogor (Bogor Agricultural University) has established a collaborative agreement with the Indonesian Ministry of State for Population and Environment and the United States Primate Research Consortium, consisting of the University of Washington Regional Primate Research Center (UW-RPRC), the Oregon Regional Primate Research Center (ORPRC), and the Bowman Gray School of Medicine at the Wake Forest University, to populate and manage a breeding facility of longtailed macaques (Macaca fascicularis) on Tinjil island, a 6 km2 island off the southern coast of West Java, Indonesia. Screening protocols have been established to select only simian retrovirus (SRV)-free animals for the colony. Animals originating in either West Java or Sumatra were individually caged and screened over a period of 3-5 months for the presence of SRV and tuberculosis. Whole blood specimens were taken from seronegative animals for virus isolation. Two months after the first screening, all negative animals were retested for SRV antibody and virus isolation. All animals remaining negative after this testing procedure and which have at least four consecutive negative TB tests were transported to the island. To date, 1,306 animals have been screened with 478 released to the island, and at least 750 babies were born on the island. Three batches of progeny of 45-50 juveniles each have been retrieved from the island, and are being used in AIDS-related research projects. © 1994 Wiley-Liss, Inc.