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Biomedical subjects

G A Casey

Publications and source records attributed to G A Casey.

At least 37 records · Page 2Linked to original sources

Major antigenic groups of rabies virus in Canada determined by anti-nucleocapsid monoclonal antibodies.

A total of 123 rabies virus isolates from various geographical areas in Canada were characterized by a panel of 43 anti-nucleocapsid monoclonal antibodies. Four major antigenic groups are found in terrestrial mammals: "Canadian Arctic" from Ontario, Quebec and the Northwest Territories; "south-eastern Georgian Bay" from Ontario; "south mid-central skunk" from Alberta, Saskatchewan and Manitoba; and "Brook's, Alberta skunk" from a restricted area in Alberta. Bat isolates can be divided into 4 major antigenic groups: "B-1" in Eptesicus fuscus from Ontario; "B-2" in a variety of bat species from British Columbia eastward into Ontario; "B-3" in Myotis spp. from Ontario and New Brunswick; and "B-4" in E. fuscus from Alberta and Saskatchewan. A single case of bat to horse transmission of rabies virus is recorded. These street isolates are compared with isolates of fixed virus. Epidemiological aspects are discussed.

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Antigenic variants of rabies virus in isolates from eastern, central and northern Canada.

Street rabies virus isolated from 51 specimens from Ontario, Quebec, Manitoba and the Northwest Territories have been typed by a panel of 36 antinucleocapsid monoclonal antibodies. Three main groups were found. The first group comprised those terrestrial mammals originating in Ontario, Quebec and the Northwest Territories. The second group was found in terrestrial mammals from Manitoba. The third heterogenous group was made up of bats from Ontario.

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Experimental rabies in skunks: effects of immunosuppression induced by cyclophosphamide.

Striped skunks (Mephitis mephitis) were inoculated with street rabies virus and immunosuppressed with several doses of cyclophosphamide. Control skunks were inoculated with street virus only. The skunks were killed in terminal stages of the disease and several tissues were collected for examination by immunofluorescence, light microscopy and viral titration. Sera collected at euthanasia from most of the principals did not contain detectable rabies neutralizing antibodies, whereas high titers occurred terminally in controls. Immunofluorescence was much more entensive in submandibular salivary glands of cyclophosphamide-treated than control skunks. Similarly, virus was isolated from this tissue more consistently and at higher titer from principals than from controls. Immunofluorescence was extensive in brains of all skunks (both groups), but virus was isolated consistently only from brains of cyclophosphamide-treated skunks. Most of the cyclophosphamide-treated skunks had very few inflammatory cells in brain and cerebrospinal ganglia. Neuronal degeneration occurred in dorsal root ganglia of both principals and controls. The results suggest that the immune response has no effect on the development of rabies-induced aggressive behavior, that the immune response may inhibit salivary gland infection and that it is not essential for the development of neuronal degeneration in dorsal root ganglia.

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Rabies virus in the salivary glands and nasal mucosa of naturally infected skunks.

Several salivary glands and the nasal mucosa of rabid skunks (Mephitis mephitis) contained rabies virus. Generally titers were high in the submandibular, moderate in the parotid and low to moderate in the zygomatic, molar and sublingual salivary glands. The nasal mucosa (glands and epithelium) contained virus at low to moderate titers that occasionally were equal to titers in brain.

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Experimental rabies in skunks: mechanisms of infection of the salivary glands.

Striped skunks (Mephitis mephitis) were inoculated into the right submandibular salivary gland with street rabies virus. They were killed at various times after inoculation and several tissues were examined by immunofluorescence and light microscopy. Right and left superior cervical, nodose and trigeminal ganglia, medulla oblongata and at least three regions of right and left submandibular salivary glands were examined by the fluorescent antibody technique. Intracerebral titrations of salivary gland suspensions were made in weanling white Swiss mice. Immunofluorescent material (inoculum) was detected in septa and connective tissue surrounding secretory units of the right submandibular gland immediately after inoculation, but otherwise antigen was not detected in either right or left submandibular glands without coincident antigen in the medulla oblongata. This occurred first on day 12 in areas of the gland remote from the inoculation site. Titers of virus were low at this time. Serum neutralizing antibodies occurred by day 7 in a few skunks. The time of development and distribution of antigen strongly suggest that, even after direct inoculation, neural networks are necessary for development of widespread infection of the salivary gland. The early occurrence of serum neutralizing antibodies in some of the skunks suggests that the immune response was activated by virus in the inoculum since immunofluorescence was not detected in any tissue at this time.

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Antigenic variants of rabies virus.

The authors studied nineteen street virus isolates from different regions of Canada using either anti-nucleocapsid and anti-glycoprotein monoclonal antibodies or cross-protection tests. This study only allowed us to recognize three groups of viruses with different nucleocapsid patterns, and no difference, as far as protection with a Pitmann-Moore vaccine is concerned, between four of these strains.

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Experimental rabies in skunks: persistence of virus in denervated muscle at the inoculation site.

Striped skunks (Mephitis mephitis) were inoculated into the denervated abductor digiti quinti muscle with street rabies virus. They were killed at various times after inoculation and several tissues were examined by immunofluorescence and light microscopy. Muscle at the inoculation site was examined electron microscopically. Rabies antigen was detected in muscle fibers first on day 7 and persisted until day 28. Light and electron microscopic lesions at the inoculation site included atrophic and degenerating muscle fibers and a few focal and regional endomysial accumulations of macrophages, lymphocytes and plasma cells. Scattered myocytes contained bodies of matrix, virions and anomalous tubular structures on electron microscopic examination. The results indicate that replication of rabies virus may occur in infected muscle fibers at the inoculation site until 28 days after exposure. This could contribute to variations in the incubation period for the first two to three months after exposure. However, the results do not support the contention that virus is contained in striated muscle cells throughout the long incubation periods.

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Experimental rabies in skunks: immunofluorescence light and electron microscopic studies.

Striped skunks (Mephitis mephitis) were inoculated into the abductor digiti quinti muscle with street rabies virus isolated from salivary glands of rabid skunks. Using the immunofluorescence technique, antigen was detected in muscle cells at the inoculation site before it was detected in the central nervous system. Neurons and their processes in nearly all regions of the brain, spinal cord, cerebrospinal ganglia, and peripheral nerves contained antigen in terminal stages of the disease. Electron microscopically, matrix (viral nucleocapsid), virions, and anomalous viral products were mainly in neuronal perikarya and dendrites, and less often in myelinated axons. Matrices, virions, and crystalloid structures were in muscle fibers at the inoculation site. Viral budding occurred on endoplasmic reticulum, neurotubules, and neuronal plasma membrane. In the brain and dorsal horn of the spinal cord, virus budded from the postsynaptic and adjacent dendritic or perikaryal plasma membrane. There was simultaneous esotropic uptake of these particles by adjacent axon terminals. The results strongly suggest that direct transneuronal transfer of virus from perikarya and dendrites to adjacent axon terminals is a mechanism in dissemination of rabies in the central nervous system of striped skunks. Variation in the length of the incubation period may be due partly to replication or virus in myocytes at the inoculation site and subsequent transfer to peripheral nerves.

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Experimental oral and nasal transmission of rabies virus in mice.

Weanling female white Swiss mice were exposed to challenge virus standard rabies virus and street virus isolates from various domestic and wild animals. Virus was given free choice as suspension or as infected mouse brain by stomach tube, by single injection of suspension into the oral cavity of unanesthetized mice, by repeated injection into the oral cavity of anesthetized mice and by single application to the external nares of anesthetized mice. Challenge virus standard virus in mouse brain suspension and a suspension of skunk salivary glands infected with street virus (titers greater than or equal to 10(6)MICLD50/0.03 ml) consistently produced high rates of infection in mice exposed intranasally, low to high rates of infection in mice exposed by forced feeding and other artificial methods of oral exposure and very low rates of infection when given free choice. Street virus isolates passaged intracerebrally in mice had titers less than or equal to 10(4.5) MICLD50/0.03 ml and rarely caused rabies in mice exposed orally or nasally by any method. The results indicate that with the isolates used, virus of high titer (greater than or equal to 10(6)MICLD50/0.03 ml) is required to consistently produce infection in mice by the nasal route and that the mucosa of the nasal cavity probably is the chief route of infection even after oral administration.

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Experimental rabies in skunks: oral, nasal, tracheal and intestinal exposure.

Striped skunks (Mephitis mephitis) were exposed to challenge virus standard rabies virus by feeding infected mouse brain in suspension or as intact brain free choice, by forced feeding of suspension, and by intranasal, intratracheal and intraintestinal instillation of suspension. All of five skunks exposed intranasally, two of five exposed intratracheally and two of ten exposed by forced feeding developed rabies. None of the skunks exposed to challenge virus standard virus, by other methods, became rabid. Most of the survivors, when challenged intramuscularly with street rabies virus at six months, developed rabies. The results indicate that the skunk is much more susceptible to challenge virus standard rabies virus given intranasally than by the other methods used. When disease occurs following oral administration, infection may be associated with prolonged contact with buccal mucosa or accidental contact with nasal mucosa. Survivors had little or no protection when challenged intramuscularly with street rabies virus.

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The mouse inoculation test in rabies diagnosis: early diagnosis in mice during the incubation period.

Brain tissue from 64 rabies suspect specimens were inoculated intracerebrally into twenty 9-12 gm adult Swiss white mice. Two mice from each specimen were killed on specific days postinoculation and examined for the presence of rabies virus by the fluorescent antibody staining technique. In this way a positive diagnosis was made in the majority of cases between postinoculation days 4 and 12 when the incubation period of these same specimens ranged between eight and 20 days.

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Demonstration of rabies antigen in salivary glands of rabies suspected animals.

Submaxillary salivary glands from 129 rabies suspected animals were studied by the following methods: a) microscopic examination of frozen sections stained by the Fluorescent antibody technique (FAT), and b) mouse infectivity test (MIT). Flourescent antibody staining of frozen sections from the salivary glands of rabid animals proved to be a satisfactory method for demonstrating rabies antigen, when compared with the mouse infectivity test.

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Skunk rabies.

In North America, the number of cases of rabies diagnosed in skunks generally exceeds that in either raccoons or foxes. Enzootic skunk rabies occurs mainly in four geographic regions: (1) southern Ontario and Quebec and upper New York State; (2) the north central United States and the Canadian provinces of Manitoba, Saskatchewan, and Alberta; (3) California; and (4) south central United States (Texas and several adjacent states). Rabies in these areas (in skunks and, to a large extent, in other terrestrial mammals) is caused mainly by three street virus variants, as determined by monoclonal antibody testing (one variant for areas 2 and 3 and separate variants for each of areas 1 and 4). Experimental studies suggest that the species specificity (e.g., raccoon vs. skunk) of enzootic rabies is due, at least partly, to differences in the pathogenicity of variants of rabies virus.

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