Browsing by Author "Blagburn, B. L."
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- Characterization of temperature-sensitive strains of Neospora caninum in miceLindsay, David S.; Lenz, S. D.; Blagburn, B. L.; Brake, D. A. (American Society of Parasitology, 1999-02)Temperature-sensitive (ts) strains of the Neospora caninum tachyzoites were selected by chemical mutagenesis and selection for growth at 32 C. Three ts strains and the parental, N. caninum wild-type strain, NC-1, were examined in the present study for their ability to cause disease in inbred BALB/c mice, outbred ICR mice, and chemically immunosuppressed ICR mice. In BALB/c mice, all 3 strains failed to induce clinical disease, whereas infection with the NC-1 strain caused central nervous system disease and death in some mice. No disease was observed in ICR mice inoculated with the 3 ts strains or the NC-I strain. All immunosuppressed ICR mice inoculated with the NC-1 strain died, whereas no immunosuppressed mice inoculated with the NCts-4 strain and only 1 of 5 mice inoculated with the NCts-8 and NCts-12 strains died. The NCts-4 and NCts-12 strains reverted to a wild-type phenotype when grown at 37 C. Vaccination of BALB/c mice with live, but not frozen NCts-8 strain tachyzoites induced significant (P < 0.05) protection following NC-1 strain challenge.
- Complete development of the porcine coccidium Isospora suis Biester, 1934 in cell culturesLindsay, David S.; Quick, D. P.; Steger, A. M.; Toivio-Kinnucan, M. A.; Blagburn, B. L. (American Society of Parasitology, 1998-06)Development from inoculated sporozoites to unsporulated oocysts of Isospora suis Biester, 1934 is described in a swine testicular (ST) cell line. Sporozoites penetrated ST cells within 1 hr postinoculation (PI). Development was initially by endodyogeny to produce binucleate type I meronts and type I merozoites. Division by endodyogeny continued during the 13-day observation period and type I merozoites were the developmental stages most abundant at observation periods >3 days PI. Mutinucleate type II meronts and type II merozoites were first observed 7 days PI. Gamonts and oocysts were present 12 days PI. Oocysts did not sporulate in vitro. The ultrastructural features of stages were similar to those that occur in the pig host.
- Examination of extraintestinal tissue cysts of Isospora belliLindsay, David S.; Dubey, Jitender P.; Toivio-Kinnucan, M. A.; Michiels, J. F.; Blagburn, B. L. (American Society of Parasitology, 1997-08)Relapse is common in immunocompetent and immunosuppressed humans infected with Isospora belli and is believed to be associated with the presence of extraintestinal stages. In the present study, we examined this important stage in an AIDS patient using histological, immunohistological, histochemical, and ultrastructural methods to better understand the development and structure of this stage and to develop better means of detecting infections. Antisera made in rabbits to Isospora suis, Toxoplasma gondii, Hammondia hammondi, Sarcocystis neurona, Neospora caninum, and Caryospora bigenetica were tested against I. belli tissue cysts in the avidin-biotin peroxidase complex (ABC) immunohistological test. Most antisera reacted positively in the ABC test at dilutions of 1:100 but not at dilutions of 1:250. Some antisera to N. caninum and H. hammondi reacted positively at dilutions of 1:1,000 in the ABC test. Most reactive antisera stained the tissue cyst wall and not the enclosed zoite. Eight histochemical tests were examined and most were nonreactive with I. belli zoites or tissue cysts. Transmission electron microscopy revealed that the tissue cyst wall was composed of granular material and was directly beneath the parasitophorous vacuole membrane. Zoites were in the center of the tissue cysts and were surrounded by fibrillar material that appeared to originate from the zoite surface. Tubulelike structures were present in the granular tissue cyst wall and in the fibrillar material that surrounded the zoite. Zoites contained a crystalloid body. New findings in the present study consisted of identifying what are probably early tissue cysts that lack a developed tissue cyst wall, demonstrating that more than 1 tissue cyst can occupy a host cell, describing the distribution of micronemes and the shedding of zoite membranes, and identifying tubular structures in the inner tissue cyst wall and inner compartment.
- Prevalence of agglutinating antibodies to Neospora caninum in raccoons, Procyon lotorLindsay, David S.; Spencer, J.; Rupprecht, C.; Blagburn, B. L. (American Society of Parasitology, 2001-10)Neospora caninum is an apicomplexan parasite that causes neonatal neuromuscular disease in dogs and abortions in cattle. Dogs are the only proven definitive host. Little is known about the prevalence of antibodies to this parasite in wildlife. Sera from 99 raccoons (Procyon lotor) were examined for agglutinating antibodies to N. caninum using the modified agglutination test employing formalin-fixed tachyzoites as antigen. Raccoons originated in Florida (n=24, collected in 1996), New Jersey (n=25, collected in 1993), Pennsylvania (n=25, collected in 1999), and Massachusetts (n=25, collected in 1993 and 1994). Ten (10%) had antibodies to AT. caninum; 9 had titers of 1:50, and 1 (1%) had a titer of 1:100. The present study indicates that raccoons have minimal exposure to N. caninum. The sera were also tested for agglutinating antibodies to Toxoplasma gondii and 46 (46%) were positive; 16 had titers of 1:50, 8 had titers of 1:100, and 22 had titers of greater than or equal to1:500.
- Vaccination of mice with Neospora caninum: Response to oral challenge with Toxoplasma gondii oocystsLindsay, David S.; Lenz, S. D.; Dykstra, C. C.; Blagburn, B. L.; Dubey, Jitender P. (American Society of Parasitology, 1998-04)Neospora caninum is a protozoan parasite that can cause severe disease in mammals. Two experiments were conducted to examine the effects of subcutaneous (s.c.) vaccination with Hank's balanced salt solution (HBSS), 1 x 10(5) N. caninum NC-1 strain tachyzoites or 1 x 10(5) Toxoplasma gondii TS-4 strain tachyzoites on challenge oral infections in mice with sporulated VEG strain T. gondii oocysts (1 x 10(3) oocysts exp. 1 and 5 x 10(3) oocysts exp. 2). An additional study, experiment 3, evaluated s.c. challenge with 2.5 x 10(3) tachyzoites of the highly virulent RH strain of T. gondii after vaccination with HBSS, NC-1 tachyzoites, or TS-4 tachyzoites. Mice vaccinated with NC-1 strain tachyzoites survived significantly (P < 0.05) longer than mice given HBSS in experiment 1, but not in experiments 2 and 3. Mice vaccinated with TS-4 strain tachyzoites survived significantly longer than HBSS-vaccinated mice in experiments 1, 2, and 3 and significantly longer than mice vaccinated with the NC-1 strain in experiments 2 and 3. Toxoplasma gondii tissue cyst numbers were significantly lower for mice vaccinated with TS-4 strain tachyzoites than mice vaccinated with HBSS or the NC-1 strain tachyzoites in experiment 1. No difference was observed in tissue cyst numbers in mice vaccinated with HBSS or NC-1 strain tachyzoites in experiment 1. No HBSS-vaccinated mice survived experiment 2, and the numbers of T. gondii tissue cysts were significantly lower for mice vaccinated with the TS-4 strain tachyzoites compared to NC-1 strain tachyzoites. No HBSS- or NC-1-vaccinated mice survived RH strain challenge in experiment 3. Results of these experiments indicate that infection with N. caninum provides some protection against fatal oral infection with T. gondii oocysts of a moderately pathogenic strain but not tachyzoites of a highly pathogenic strain. The protection provided bq N. caninum is much less than that provided by previous exposure to T. gondii, and the numbers of tissue cysts in the brains of mice are not significantly (P > 00.5) lowered.