A cross sectional survey for B virus antibody in a colony of group housed rhesus macaques

A systematic sampling technique was used in combination with a highly sensitive and specific ELISA to provide unbiased age-specific prevalence estimates of B virus antibody in rhesus monkeys housed in three different outdoor breeding corrals. Among 146 sampled monkeys, 97% of animals 2.5 years and older were seropositive, while only 22% of younger animals were seropositive. Neither gender nor social dominance ranking were predictive of B virus antibody status. The strong age association was not inconsistent with hypothesized venereal transmission of B virus. Improvements in the epidemiologic understanding of B virus are necessary to assist efforts to eradicate this agent from breeding colonies of rhesus monkeys.     

Excretion of B virus in monkeys and evidence of genital infection

3 different types of observation all demonstrated that B virus (Herpesvirus simiae) infection of monkeys was not confined to the mouth but was also a genital infection. (1) Latent B virus was reactivated in a seropositive female monkey, which was immunosuppressed with antilymphocyte globulin, and infectious virus was excreted in the genital tract. (2) During an epizootic in a breeding colony, B virus was isolated from 4 genital and 3 oral sites as well as from a skin lesion. (3) In cultures of sensory nerve ganglia taken from seropositive monkeys, B virus was recovered more frequently from ganglia subserving the genital region than the oral region.     

Prevalence of herpes B virus genome in the trigeminal ganglia of seropositive cynomolgus macaques

Herpes B virus infection is almost asymptomatic in macaques (Macaca spp.), which are the natural hosts of this pathogen, but is the cause of high mortality in humans. Reactivation of the latent virus in the trigeminal ganglia (TG) results in the shedding of infectious particles into the oral mucosal membrane. Saliva contaminated with the reactivated virus from the ganglia of the natural host is considered to be important for viral transmission to humans and other monkeys. In the present study, we investigated the prevalence of the herpes B virus genome in the left and right TG of seropositive asymptomatic cynomolgus macaques. The latent virus genome was detected using a polymerase chain reaction and microplate hybridization assay. We found that the virus DNA was present in one or both TG of 12 of the 30 macaques (40%) tested, with the virus being detected from both TG in five of the 12 macaques and from a single TG in the remaining seven.     

B-virus specific-pathogen-free breeding colonies of macaques (Macaca mulatta): retrospective study of seven years of testing.

BACKGROUND AND PURPOSE: National Institutes of Health's Division of Comparative Medicine has sponsored a multi-institutional program for the establishment of specific-pathogen-free (SPF) macaque colonies. B virus (Herpesvirus simiae, Cercopithecine herpesvirus type 1) has been targeted in this surveillance. Participating institutions have established individual timetables for frequency of testing and types of monitoring and husbandry techniques, all with the common goal of producing pathogen-free monkeys for research. The greatest biosecurity threat to the program comes from failure to detect seronegative latent infections, either in first-year macaques or macaques introduced in subsequent years, although these are supposed to operate as closed colonies. METHODS: From January 1990 through December 1996, we screened macaques for B virus, using enzyme-linked immunoabsorbant assay (ELISA) and Western blot analysis. RESULTS: During the first year, 1,097 macaques from six colonies were tested, and 88.4% tested negative for B virus. During the seventh year, 1,843 were tested, of which 99.7% tested negative. Seropositive macaques were detected as late as the seventh year. CONCLUSIONS: An aggressive program to establish an SPF colony of captive breeding macaques can be effective in reducing the risk of B-virus exposure.     

A serological survey of simian virus 40 in monkeys

A serological survey of simian virus 40 (SV 40) was conducted by an immune adherence hemagglutination (IAHA) test in breeding monkeys. Of a total of 356 monkeys tested, 168 (47.2%) were seropositive. All 168 seropositive monkeys were detected from 224 monkeys which were bred or kept in Japan for a long time. In contrast, none of the 132 monkeys which were newly imported from Southeast Asia was seropositive. If a comparison was made in the same breeding place, the positive rate of 80.4% (111/138) of Japanese monkeys was significantly (P less than 0.01) higher than the 59.5% (25/42) among rhesus monkeys. The positive rate and the IAHA titers were higher in older age group (greater than 5 years) but similar in male and female. These results indicated that SV 40 was highly prevalent among breeding monkeys in Japan.     

Antibodies against Pasteurella multocida in snow geese in the western Arctic.

To determine if lesser snow geese (Chen caerulescens caerulescens) are a potential reservoir for the Pasteurella multocida bacterium that causes avian cholera, serum samples and/or pharyngeal swabs were collected from > 3,400 adult geese breeding on Wrangel Island (Russia) and Banks Island (Canada) during 1993-1996. Pharyngeal swab sampling rarely (> 0.1%) detected birds that were exposed to P. multocida in these populations. Geese with serum antibody levels indicating recent infection with P. multocida were found at both breeding colonies. Prevalence of seropositive birds was 3.5% at Wrangel Island, an area that has no recorded history of avian cholera epizootics. Prevalence of seropositive birds was 2.8% at Banks Island in 1994, but increased to 8.2% during 1995 and 1996 when an estimated 40,000-60,000 snow geese were infected. Approximately 50% of the infected birds died during the epizootic and a portion of the surviving birds may have become carriers of the disease. This pattern of prevalence indicated that enzootic levels of infection with P. multocida occurred at both breeding colonies. When no avian cholera epizootics occurred (Wrangel Island, Banks Island in 1994), female snow geese (4.7%) had higher antibody prevalence than males (2.0%).     

Isolation of a simian immunodeficiency virus related to human immunodeficiency virus type 2 from a west African pet sooty mangabey.

Two of 25 healthy pet sooty mangabey (SM) monkeys (Cercocebus atys) living in West Africa were seropositive by immunoblot when surveyed for antibody to simian immunodeficiency virus of macaques (SIVmac). SIVsmLIB1 was isolated from one of the pet sooty mangabeys. Nucleotide sequence data showed that this isolate is a member of the SIVsm/human immunodeficiecy virus type 2 (HIV-2)/SIVmac group of primate lentiviruses. Furthermore, sequence comparisons revealed extensive genetic diversity among SIVsm isolates similar to that observed previously in SIV isolates from naturally infected African green monkeys. These observations provide additional evidence for monkey-human cross-species transmission of SIVsm as the source of HIV-2 infection of human.     

A study of the functional capabilities of human neonatal lymphocytes for in vitro specific antibody production

The ability of neonatal B and T cells to participate in the in vitro production of anti-influenza virus antibody was studied. Peripheral blood mononuclear cells from nearly all normal adults produce anti-influenza virus antibody when stimulated in vitro with type A influenza virus. Cord blood mononuclear cells, however, consistently failed to do so. Using Epstein-Barr virus activation or coculture with irradiated adult T cells in the presence of influenza virus to identify precursor B cells for anti-viral antibody production, newborns were found to have a decreased number of influenza-specific B cells as compared with adults. Thus, a paucity of precursor B cells for anti-influenza virus antibody was one factor contributing to the absent in vitro antibody response. Additional studies were undertaken to investigate the capabilities of newborn T cells in the in vitro response to influenza virus. Newborn T cells failed to proliferate when cultured with influenza virus. Irradiated newborn T cells were, however, able to provide help for specific antibody production in influenza virus-stimulated cocultures with allogeneic adult B cells, and newborn T cells proliferated when stimulated with alloantigens; their helper function in allogeneic coculture was, thus, likely mediated by T cells stimulated by alloantigens rather than by influenza virus. In the absence of T cell irradiation, no antibody was produced in cocultures of adult B cells and neonatal T cells, at least in part as the result of a radiosensitive suppressor T cell. Suppression in influenza virus-stimulated allogeneic cocultures was also observed with normal adult T cells and is, therefore, a property of both newborn and adult T cells. Thus, allogeneic help and suppression can both be manifested in exogenous antigen-stimulated allogeneic cocultures. In addition, both of these allogeneic effects can be mediated by neonatal T cells, indicating that these functions are present at the time of birth and do not require previous exogenous antigenic exposure for expression.     

Seroprevalence of B virus (Herpesvirus simiae) antibodies in a naturally formed group of rhesus macaques

Eighty-two percent of a group of rhesus monkeys removed from Cayo Santiago were seropositive for B virus (Herpesvirus simiae) antibodies. Similar results were obtained from the Cayo Santiago macaque population two decades ago and from feral Indian rhesus monkeys. Thus it is likely that B virus has been enzootic in the Cayo Santiago population since 1938, when the colony was established with stock imported from India.     

Activation of B virus (Herpesvirus simiae) in chronically immunosuppressed cynomolgus monkeys.

Three of 14 cynomolgus monkeys given the highest dose of an immunosuppressive drug in a 6-month toxicology study developed B virus (Herpesvirus simiae) oral lesions after 3 months of dosing. This necessitated early removal of all high-dose monkeys from the study due to concerns related to B virus. The incidence and severity of parasitic (Oesphagostomum sp.) lesions of the large intestine were also increased in high-dose animals. Both B virus and Oesophagostomum are enzootic in macaques, and the lesions caused by them were considered secondary to chronic immunosuppression caused by the highest dose of the test compound. Evidence of immunosuppression included decreased lymphocyte counts (B-cells; CD2 and CD8 T-cells), histopathologic evidence of lymphoid suppression, and serum-induced inhibition of lymphocyte mitogen responses. Pathogenesis of the B virus was apparently associated with both activation of latent virus as well as transmission of active virus. Approaches for virologic monitoring of primates and for ensuring optimal safety for primate handlers are discussed.     

猴B病毒相关抗体及B病毒抗体检测的比较研究

本文比较了用ＨＳＶ－１（ＩＥＡ）和Ｂ病毒（ＩＦＡ）的方法检查猴Ｂ病毒相关抗体和Ｂ病毒抗体的结果：ＩＥＡ检查出的Ｂ病毒相关抗体阴性猴,经ＩＦＡ检查,全部为Ｂ病毒抗体阴性。用ＩＦＡ检查了Ｂ病毒相关抗体阴性的恒河猴,在单笼隔离饲养六个月后,有９８．３％的动物Ｂ病毒相关抗体仍为阴性,表明ＩＲＡ的阴性结果有较好的一致性。本文讨论了建立无Ｂ病毒感染猴群的可行性。     

Behavioral research in breeding colonies of Old World monkeys.

The existence of an active behavioral research program using animals in primate breeding colonies was considered to be not only a compatable multiple use of animals, but a way of materially improving the management and efficiency of the breeding colonies. In colonies of monkeys specifically established for behavioral research programs directed at the examination of social relationships, incidental breeding resulted in levels of reproductive success equivalent to or greater than that normally experienced in colonies devoted entirely to breeding. Behavioral research revealed patterns of seasonality, fostering, kidnapping, and infant care which would otherwise have escaped notice and which would significantly influence culling and management choices in a breeding colony. Many young males and certain low ranking adult males actively contributed to reproduction. Females born and reared in the colonies were the most productive, exceeding wild born or other introduced females in reproductive efficiency. Specific recommendations for establishing, expanding and culling of nonhuman primate breeding colonies were derived from the behavioral research.     

Acquisition and Transmission of two Begomoviruses by the B and a non-B Biotype of Bemisia tabaci from Zhejiang, China

Acquisition and transmission was studied of Tomato yellow leaf curl China virus (TYLCCNV) and Tobacco curly shoot virus (TbCSV) by the B and a non‐B biotype (China‐ZHJ‐1) of Bemisia tabaci from Zhejiang, China. The frequency of TYLCCNV and TbCSV detection by PCR in whitefly adults increased with increasing length of feeding on virus‐infected plants. The virus DNA was detected by PCR in 40% of the B biotype adults tested after a period of 30 min access to infected plants and in all adults after a 12‐h period of access. All ZHJ‐1 adults acquired TYLCCNV and TbCSV after a 48‐h period of access to the virus‐infected plants. Viruliferous B and ZHJ‐1 adults retained TYLCCNV DNA for their entire life when placed on healthy cotton plants. Viruliferous ZHJ‐1 adults retained TbCSV DNA for their entire life when placed on healthy cotton plants but the B biotype adults did not. Transmission of TYLCCNV was achieved with one B or ZHJ‐1 adult per plant, and the probability of transmission reached 100% when the number of adults was increased to 10 per plant. The efficiency for TYLCCNV transmission to healthy plants by adults of both B and ZHJ‐1 was much higher than that for TbCSV.     

Borrelia burgdorferi Sensu Lato in an Endemic Environment: Wild Sika Deer (Cervus nippon yesoensis) with Infected Ticks and Antibodies

Ticks and blood samples were collected from wild sika deer (Cervus nippon yesoensis) during a hunting season (August to October) of 1991 at a selected location in Hokkaido, Japan. Ixodes persulcatus (adult and nymph) and I. ovatus (adult) were the common ticks on sika deer. Spirochetes were detected in the midgut of the ticks by the indirect peroxidase-conjugated antibody staining method and by dark-field microscopy after cultivation. By the reactive pattern of monoclonal antibodies, isolates were considered to belong to Borrelia garinii or B. japonica. In an antibody test, the percentage of seropositive deer was 69.0%. Most of the adult sika deer were positive for antibodies to the spirochetes. There are significant age-dependency in antibody level and seropositive rate. The surveillance of deer should be valuable in monitoring the transmission risk of B. burgdorferi sensu lato in nature.     

Cell-to-cell transmission of human immunodeficiency virus type 1 in the presence of azidothymidine and neutralizing antibody.

Very few peripheral blood lymphocytes of seropositive individuals are presumably actively infected with human immunodeficiency virus type 1 (HIV-1). During coculture of lymphocytes of a seropositive individual with mitogen-stimulated normal peripheral blood lymphocytes, the number of infected cells becomes amplified such that detectable HIV-1 is produced. We report here that in addition to transmission by extracellular virus, cell-to-cell transmission is responsible for spreading HIV-1 infection from infected to uninfected cells. Azidothymidine and virus-neutralizing antibody had no effect on cell-to-cell transmission of HIV-1. Monoclonal antibodies to the CD4 receptor, but not to the CD3 receptor, prevented cell-to-cell transmission, which suggests that CD4 receptor-mediated cell fusion is involved in cell-to-cell transmission. Spread of infection in a cell-to-cell manner may be important in development of drug therapies for HIV-1 infection.     

Experimental infection of cynomolgus monkeys with simian parvovirus.

Simian parvovirus is a recently discovered parvovirus that was first isolated from cynomolgus monkeys. It is similar to human B19 parvovirus in terms of virus genome, tropism for erythroid cells, and characteristic pathology in natural infections. Cynomolgus monkeys were infected with simian parvovirus to investigate their potential usefulness as an animal model of human B19 parvovirus. Six adult female cynomolgus monkeys were inoculated with purified simian parvovirus by the intravenous or intranasal route and monitored for evidence of clinical abnormalities; this included the preparation of complete hematological profiles. Viremia and simian parvovirus-specific antibody were determined in infected monkeys by dot blot and Western blot assays, respectively. Bone marrow was examined at necropsy 6, 10, or 15 days postinfection. All of the monkeys developed a smoldering, low-grade viremia that peaked approximately 10 to 12 days after inoculation. Peak viremia coincided with the appearance of specific antibody and was followed by sudden clearance of the virus and complete, but transient, absence of reticulocytes from the peripheral blood. Clinical signs were mild and involved mainly anorexia and slight weight loss. Infection was associated with a mild decrease in hemoglobin, hematocrit, and erythrocyte numbers. Bone marrow showed marked destruction of erythroid cells coincident with peak viremia. Our findings indicate that infection of healthy monkeys by simian parvovirus is self-limited and mild, with transient cessation of erythropoiesis. Our study has reproduced Koch's postulates and further shown that simian parvovirus infection of monkeys is almost identical to human B19 parvovirus infection of humans. Accordingly, this animal model may prove valuable in the study of the pathogenesis of B19 virus infection.     

Herpesvirus simiae (B virus) antibody response and virus shedding in experimental primary infection of cynomolgus monkeys.

Four cynomolgus monkeys (Macaca fascicularis) were inoculated in the lips and tongues with B virus. Virus shedding and antibody responses were monitored for up to 50 days postinfection. Virus was isolated from the oral cavities of all monkeys at 6 days postinfection despite the absence of observable lesions. Virus was not isolated from genital swabs or serum. Antibodies to both B virus and herpes simplex virus were detected by neutralization between days 8 and 12. Virus-specific IgM and IgG antibodies were measured by antibody capture radioimmunoassay. IgM was first detected on day 6; by contrast, IgG did not appear until day 12. Antibodies reactive in a competitive radioimmunoassay appeared by day 12 and peaked at 30 to 40 days postinfection. This study provides data on which to base the diagnosis of primary B virus infection in cynomolgus monkeys.     

Development of a SCAR marker for the analysis of B chromosome presence in Partamona helleri (Hymenoptera, Apidae)

Chromosomes in hymenopteran insects cannot currently be analysed in adult individuals. The only available cytogenetic techniques need to be performed in larvae. Here we develop and implement a SCAR (Sequence Characterized Amplified Region) marker, associated with B chromosomes in the bee Partamona helleri, which has proven to be very useful to reveal B chromosome presence in adults from natural populations. The marker was tested in ten different colonies simultaneously analysed by both molecular (ten adults per colony) and cytogenetic (20 larvae per colony) techniques. The presence of the SCAR marker always showed the same pattern as B chromosome presence: both were present or absent in all individuals from a same colony, or both were present in only part of the individuals from a same colony. This molecular marker is thus a useful tool for analysing new aspects of this B chromosome system such as B frequency and geographical distribution, B transmission, or B effects in adult individuals.