Hippocampal neuronal atrophy occurs in rhesus macaques following infection with simian immunodeficiency virus.

There is strong evidence that patients with AIDS have loss of cortical neurons. In this study we have examined the hippocampus of rhesus monkeys infected with simian immunodeficiency virus (SIV) to determine whether neuronal damage occurs in this model of human AIDS and to investigate its time course. Twenty-eight infected monkeys (23 young [< 9 years] and five elderly [> 16 years]) were compared with 11 controls (six young and five elderly). Numbers of nucleolated neurons per unit area of section and mean pyramidal cell diameters were measured in each CA sub-field of each animal. There was neuronal atrophy in all regions examined, as early as 3 months following inoculation. An initial apparent increase in neuronal density at 3 months did not reach statistical significance. In younger animals, however, there was a later, significant association between the reduction in neuronal density and duration of infection. Elderly animals were only examined at a single, early time point. These results show that there is neuronal pathology following infection with SIV and that there is probably subsequent neuronal death.

The effect of growth rate and haemin on the virulence and proteolytic activity of Porphyromonas gingivalis W50.

Porphyromonas gingivalis strain W50 was grown under haemin-limitation and haemin-excess conditions in a chemostat at pH 7.5. The maximum specific growth rate (mumax) was determined at both haemin concentrations (mumax = 0.236 +/- 0.052 and 0.271 +/- 0.039 h-1, respectively). This enabled dilution rates to be adjusted so that the virulence and enzyme activity of haemin-limited and haemin-replete cells could be compared at identical relative growth rates (murel) of 0.25, 0.50 and 0.75 of their respective mumax. The data showed that the fastest growing cells were significantly more virulent than those grown more slowly, irrespective of haemin concentration. However, at each growth rate tested, cells grown under haemin-excess conditions were always more virulent than haemin-limited cells. Trypsin-like enzyme activity of whole cultures was also greater at each growth rate under haemin-excess conditions while, conversely, collagenolytic activity was generally higher in haemin-limited cultures. Thus, although growth rate had an effect on the virulence and enzyme activity of P. gingivalis, the availability of haemin for growth was the most significant factor.

A comparison of aerosol and intragastric routes of infection with Listeria spp.

Aerosol infection (AI) of Porton outbred mice with Listeria species, exhibiting varying degrees of virulence, was compared with gastric intubation (GI) on the basis of numbers of deaths, 50% lethal dose (LD50) and pattern of listerial infection. The AI route appeared to be more sensitive, efficient and consistent than GI in that it required 10(5) fewer micro-organisms to obtain infection and death then ensued within 4 days, with GI deaths usually occurring on day 7. All the virulent strains tested caused 100% mortality by AI, while virulent and avirulent strains were indistinguishable by GI. Bacterial counts in the livers and spleens of infected mice were consistent with the relative virulence of the infectious agent using AI but not in GI mice. There were higher numbers of micro-organisms and more widespread lesions in the organs of AI mice than in GI. Results indicate that AI is an accurate in vivo indicator of virulence in listeria and using AI, bacterial counts in the liver and spleen could replace LD50 tests, thereby reducing the number of animals required for in vivo virulence testing.

Different patterns of neuropathological disease in rhesus monkeys infected by simian immunodeficiency virus, and their relation to the humoral immune response.

The brains of 21 rhesus monkeys inoculated with SIVMAC251 were examined after intervals ranging from 3 to 27 months and compared with five uninoculated controls. Eighteen animals became infected and individually exhibited several distinct patterns of disease. Nine (50%) had largely intramural leptomeningeal venous infiltrates (LMVI) without multinucleate giant cells (MGC) or foamy macrophages. Three (17%) had only MGC lesions, involving the cerebral parenchyma. One had both patterns and five (33%) neither. The controls had sparse and tiny LMVI only, similar to three inoculated animals that did not become infected. Immunohistochemistry showed the predominance of T and B lymphocytes in LMVI and choroid plexus mononuclear lesions but a predominance of macrophages over lymphocytes in the MGC lesions. Specific disease patterns differed in their association with the humoral immune response. Animals with LMVI were all hypergammaglobulinaemic when killed compared to pre-inoculation levels, and the size of the change in serum immunoglobulin concentration was positively correlated with a quantitative index of LMVI density. Furthermore, their post-mortem lymph node histology was hyperplastic. In contrast, animals found at autopsy to have MGC brain lesions were hypogammaglobulinaemic compared to preinoculation. The results are consistent with two phases in SIV-associated disease: one characterized by LMVI and hypergammaglobulinaemia and another featuring MGC and hypogammaglobulinaemia.

A comparison of the efficacy of azithromycin and clarithromycin in oral therapy of experimental airborne Legionnaires' disease.

The activities of two new macrolides, azithromycin and clarithromycin, were compared in an aerosol-infected guinea-pig model of legionnaires' disease. The results of this study indicate that a low oral dose of azithromycin (3.6 mg/kg) administered once daily gives 100% survival in Legionella pneumophila-infected animals. An eight-fold higher dose of clarithromycin (28.8 mg/kg) given twice-daily was required to achieve the same effect. Similarly, azithromycin was more effective than clarithromycin in preventing pyrexia and in reducing numbers of bacteria and lesions in the lung.

Influence of feeding patterns on the artificial infection of laying hens with Salmonella enteritidis phage type 4.

Salmonella enteritidis phage type 4 was recovered significantly more frequently from the crops of birds which had been denied food for 24 hours than from birds allowed food ad libitum. There was, however, no difference in its isolation rate from tissues. Within one hour of infection, S enteritidis could be recovered from a variety of tissues, including the oviduct, of a small proportion of the infected birds.

Studies on the specificity of the vaccine effect elicited by inactivated simian immunodeficiency virus.

Inactivated, partially purified simian immunodeficiency virus (SIVmac) protected macaques from intravenous challenge with homologous and heterologous strains of SIV that had been grown on human cells but no protection against challenge with monkey peripheral blood mononuclear cell-grown SIVmac was afforded. Human immunodeficiency virus type 1 prepared in an analogous way to the SIVmac vaccine on the C8166 human T cell line protected macaques against challenge with human cell-grown SIVmac. These results suggest that protection may be mediated by xenoimmunization with the vaccine cell substrate proteins. All vaccinated macaques had anti-cell antibodies. Major reactivity to MHC class I antigens was found as well as to a 70-kD protein detectable only under nonreducing conditions.

Serological responses of chickens experimentally infected with Salmonella enteritidis PT4 by different routes.

Commercially reared chickens were challenged with Salmonella enteritidis phage-type (PT) 4 by aerosol, or via the conjunctiva. Inhalation of 2.9 x 10(2) or 4.2 x 10(3) S. enteritidis resulted in the production of IgG antibodies to the lipopolysaccharide (LPS) of S. enteritidis PT4. When the aerosol inoculum was increased to 2.4 x 10(5) bacteria per bird the antibody produced were predominantly of the IgM-class. Chickens challenged with 10(3) S. enteritidis PT4 via the conjunctiva mounted only a poor immune response. Increasing the challenge dose to 10(8) S. enteritidis resulted in the production of high-titre serum antibodies of both the IgG and IgM classes. Results from this study suggest that aerosols containing small numbers of S. enteritidis PT4 might be responsible for intraflock infection of poultry.

Infection of laying hens with Salmonella enteritidis PT4 by conjunctival challenge.

The direct administration of either 103 or 108 cells of Salmonella enteritidis phage type 4 on to the conjunctiva of laying hens resulted in systemic infection. The bacterium was isolated from a variety of tissues, including the ovary and oviduct, and it was excreted in faeces for at least 27 days after infection.

Pathological changes in the reproductive tract of male rhesus monkeys associated with age and simian AIDS.

The pathological changes associated with ageing and simian immunodeficiency virus (SIV) infection in groups of immature, adult and ageing Rhesus monkeys were studied. Eighty three per cent (5 of 6) of uninfected ageing animals had hyperplasia of the prostate, 33 per cent (2 of 6) had mild prostatitis and in 66 per cent (4 of 6) there were calcified concretions in the seminal vesicles. The testes were normal and showed active spermatogenesis. In the SIV-infected animals, two types of lesion occurred; the most common, in 81 per cent (18 of 22 monkeys), was the presence of focal lymphoid infiltrations in the epididymis, prostate or seminal vesicles. The other was hypospermatogenesis (23 per cent, 4 of 17) with degeneration of seminiferous tubules. Immunocytochemical staining demonstrated that the lymphoid masses contained approximately equal numbers of B and T lymphocytes, but the majority of diffusely scattered cells were T lymphocytes. Staining for SIV antigen identified small numbers of positive lymphocytes and macrophages in all tissues.

Interstitial pneumonia in simian immunodeficiency virus infection.

Interstitial pneumonia unrelated to Pneumocystis carinii or other infections was observed histopathologically in 5 of 25 rhesus monkeys infected with simian immunodeficiency virus (SIV). The predominant lesion was lymphocytic infiltration of interalveolar septa and hyperplasia of peribronchial and perivascular lymphoid tissue. Immunohistochemical staining using a panel of antibodies against human T and B lymphocytes, macrophages, and immunoglobulins showed that peribronchial aggregates and interstitial infiltrates were predominantly B cells, whereas perivascular masses consisted mainly of T cells. One animal with a primary B-cell lymphoma of the spinal cord had secondary plasmacytoid lymphomatous nodules throughout the lung which were accompanied locally by reactive B-cell lymphoid follicles. Another animal also had large areas of diffuse alveolar fibrosis and epithelial metaplasia to a bronchiolar type. In two monkeys, branches of the pulmonary arteries showed intimal proliferation and organizing occlusive thrombi, some of which were mineralized.

Comparison of the pathogenicity for pregnant sheep of Rift Valley fever virus and a live attenuated vaccine.

A live attenuated mutant of Rift Valley fever virus, MV P12, was previously shown to be non-pathogenic in young lambs, but capable of producing protective immunity. The studies reported here show that the abortion in sheep caused by an infection with virulent virus is the result of necrosis of the maternal villi and cotyledons arising from an acute inflammation of the maternal caruncles. Pregnant ewes infected with the attenuated mutant virus MV P12 showed none of these lesions in the placenta and gave birth to healthy lambs. Colostrum from ewes infected with MV P12 virus was able to induce protective immunity in the offspring. These data along with previously published results suggest that the mutant virus MV P12 is an excellent candidate for use as a live attenuated veterinary vaccine.

Airborne infection of laying hens with Salmonella enteritidis phage type 4.

Hens were exposed to small-particle aerosols containing different concentrations of Salmonella enteritidis phage type 4. They developed a systemic infection and some birds were still excreting the organism in the faeces when killed 28 days after infection. S enteritidis was present for a similar period in a wide range of alimentary tract issues and in the ovary and oviduct.

Intrarectal challenge of macaques vaccinated with formalin-inactivated simian immunodeficiency virus.

Macaques can be protected from intravenous infection with simian immunodeficiency virus (SIV) by vaccination with chemically inactivated virus. However, protection against infection via a mucosal surface has not been demonstrated. We vaccinated four rhesus macaques with formalin-inactivated SIV given intramuscularly. These monkeys, which had remained virus free for 10 months after intravenous challenge with SIV, were given a further dose of vaccine and together with four unvaccinated controls were challenged intrarectally with SIV. Subsequently, virus was isolated from all control animals on five successive occasions, but the vaccinated animals remained free of virus. Proviral DNA could not be detected in peripheral blood mononuclear cells from the vaccinated animals. Preliminary data indicate that vaccinated animals make a local antibody response.

Transmission studies with simian immunodeficiency virus of macaques; persistent infection of baboons.

The host range of SIVmac was investigated in three monkey species. Blood-borne and cell-adapted virus inocula obtained from a rhesus macaque infected with SIVmac251 were compared. African green monkeys were not susceptible to infection, whereas baboons and rhesus macaques became persistently infected and showed similar patterns of seroconversion. However, in contrast to the macaques, no clinical or histopathological evidence of disease was seen in the baboons 2 years after virus inoculation. Thus baboons could be used as an alternative to macaques in vaccine development studies with this particular isolate of SIVmac. Furthermore, this system may be useful for the investigation of factors responsible for disease progression.

Chronic pancreatitis and biliary fibrosis associated with cryptosporidiosis in simian AIDS.

Two Rhesus monkeys infected with simian immunodeficiency virus for 15 and 24 months developed generalized oedema and one became jaundiced. At necropsy, the liver and pancreas were hard and irregular and the gall bladder was thickened. Histopathological examination showed extensive fibrosis of the pancreas, loss of exocrine acini and marked proliferation of ductules. Numerous cryptosporidia were present on the duct epithelium. The liver of both animals had widespread cirrhosis, bile duct proliferation and cholangitis. Cryptosporidia were found in many bile ducts and on the hyperplastic gall bladder epithelium. Lymph nodes and spleen of both animals showed depletion of cortical and paracortical elements characteristic of advanced immunodeficiency virus infection.

Salmonella enteritidis PT4 infection in specific pathogen free hens: influence of infecting dose.

The period during which specific pathogen free hens, infected by direct introduction into the crop of either 10(3), 10(6) or 10(8) cells of Salmonella enteritidis PT4, excreted the organism in faeces was closely related to the size of the inoculum, with the birds excreting for mean periods of 3.4, 16.4 and 36.8 days, respectively. The production of either IgG or IgM was also dose related with the birds which received 10(3) cells having lower antibody levels than those in the other two groups. In contrast, there was no relationship between the contamination of egg contents and either antibody status, faecal excretion, or the dose administered.

Phenotypic analysis of malignant lymphoma in simian immunodeficiency virus infection using anti-human antibodies.

Primates infected with simian immunodeficiency virus (SIV) develop a condition similar to the human acquired immunodeficiency syndrome (AIDS). The close resemblance between the simian acquired immunodeficiency syndrome (SAIDS) and the human disease has led to the widespread use of SIV-infected monkeys as an animal model in the study of acquired immunodeficiency. We have investigated the use of standard anti-human antibodies for the immunohistochemical analysis of formalin-fixed, paraffin-embedded tissues from monkeys with SAIDS. With the exception of antibodies UCHL1 (CD45RO), MT1 (CD43), 4KB5 (CD45RA), and Ber H2 (CD30), our routine (human) lymphoma panel of markers worked successfully on the animal tissues. Using the anti-human antibodies, we were able to analyse the phenotypes of two cases of malignant lymphoma arising in a study group of 26 SIV-infected rhesus monkeys. Both of the cases stained with the antibodies WR16 (CD45RA) and L26 (CD20), and the B-cell lineage of the lymphomas was confirmed by the detection of IgA lambda immunoglobulin expression in one case, and IgM heavy chain in the other. We therefore report the successful use of anti-human antibodies in the immunohistochemical analysis of lymphomas arising in non-human primates infected with SIV.

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.

Pneumocystis carinii pneumonia in simian immunodeficiency virus infection: immunohistological and scanning and transmission electron microscopical studies.

Pneumocystis carinii pneumonia occurred in 6 of 17 rhesus monkeys infected with simian immunodeficiency virus and was studied by immunohistochemistry and by scanning and transmission electron microscopy. A monoclonal antibody/streptavidin-biotin-peroxidase staining method was highly sensitive for detecting the organisms in small, early lesions and was much more sensitive and specific than traditional silver impregnation methods. Reprocessing of paraffin wax-embedded lung tissue for scanning electron microscopy and use of a video printer to produce a photographic montage of light microscopic lesions allowed the same areas of tissue to be examined and compared by both methods. The ultrastructural morphology of P. carinii in the rhesus monkey was identical to that in man, as were the histological and electron microscopic lesions, including pulmonary fibrosis. Trophozoites were seen attached to alveolar type I epithelium mainly by intimate apposition to the plasma membrane, but scanning electron microscopy also showed attachment by elongated filopodia. Few macrophages were present in infected alveoli, and though phagocytosis followed by digestion of P. carinii trophozoites was observed, it appeared to occur at a very low level.