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  • Author or Editor: D. S. Lindsay x
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SUMMARY

Seven monoclonal antibodies (mab) generated against sporozoites of Eimeria bovis were tested for reactivity against immature and mature first-generation meronts, sexual stages, and oocysts in tissues from experimentally infected calves by use of an avidin-biotin peroxidase complex (abpc) immunohistologic test. Three of the 7 mab reacted in the abpc test. One of these, mab-4FB4, reacted only with mature E bovis meronts. The other 2 mab, mab-2AE7 and mab-4AD7, reacted with all the developmental stages of E bovis tested. Asexual stages and sexual stages of E tenella from chickens and E papillota from mice also were examined in the abpc test. Monoclonal antibodies mab-2AE7 and mab-4AD7 reacted with all stages of these eimerian protozoa. None of the other 5 mab reacted with these parasites. Results of this study suggested that antigens are shared among the asexual and sexual stages of several diverse Eimeria species.

Free access
in American Journal of Veterinary Research
in Journal of the American Veterinary Medical Association

Summary

Carcasses from 18 Holstein cows from Nevada, California, and Oregon were condemned because of grossly visible sarcocysts of Sarcocystis hirsuta. Sarcocysts were white to dull white, 2 to 7 mm long, and approximately 1 mm wide. Sarcocysts were most numerous in muscles of diaphragm, flank, and limbs and were able to infect laboratory-raised cats.

Free access
in Journal of the American Veterinary Medical Association

Abstract

Objective

To examine cross-reactivity among Neospora caninum and closely-related apicomplexans.

Design

Sera from animals were examined for antibody production to N caninum and cross-reactivity to Toxoplasma gondii.

Animals

Cattle were experimentally infected with 3 tissue cyst-forming protozoan parasites N caninum, T gondii, and Sarcocystis sp, and calves were monospecifically inoculated with the intestinal coccidia, Eimeria bovis and Cryptosporidium parvum. Similar studies were done in laboratory rabbits inoculated with N caninum, T gondii, Hammondia hammondi, and Sarcocystis sp. Additionally, sera were obtained from ewes, lambs, goats, sows, cats, rats, and mice inoculated with N caninum tachyzoites.

Procedure

The indirect fluorescent antibody (IFA) and ELISA antibody tests (cattle only) were used to examine reactivity to N caninum; the modified direct agglutination, Sabin-Feldman dye, and IFA tests were used to evaluate reactivity to T gondii.

Results

Serologic cross-reactivity among N caninum, T gondii, and Sarcocystis sp was none or minimal by the IFA test. There was some reactivity to N caninum by the use of ELISA in cattle inoculated with Sarcocystis sp.

Conclusions

The IFA test for N caninum was specific for the diagnosis of neosporosis in animals.(Am J Vet Res 1996;57:329-336)

Free access
in American Journal of Veterinary Research

Summary

Three Jersey cows were inoculated sc and IM with 26 million Neospora caninum tachyzoites at 129 (cow 1), 126 (cow 2), and 81 (cow 3) days after mating. Cows remained clinically normal for at least 1 month after inoculation of N caninum. Cow 1 was euthanatized 32 days after inoculation because of gangrenous mastitis. Cow 1 had a live fetus with no gross lesions; however, microscopic lesions were seen in the fetus and consisted of severe nonsuppurative necrotizing encephalitis of the cerebral white matter. Neospora caninum was identified in lesions by staining with anti-N caninum serum in an immunohistochemical test, by bioassays in mice, and by inoculation of bovine monocyte cultures with fetal tissue homogenate. Neither N caninum nor lesions were associated with infection with the protozoon identified in tissues of cow 1. Cows 2 and 3 aborted small autolysed fetuses 101 and 74 days, respectively, after inoculation with N caninum; the fetuses and attached placenta were unsuitable for laboratory investigations. Cows 2 and 3 remained clinically normal 4 months after abortion. Results of this study indicated that N caninum can be transmitted transplacentally in cattle.

Free access
in Journal of the American Veterinary Medical Association

SUMMARY

Neospora caninum-induced abortion is a major production problem in the dairy cattle industry in the United States and worldwide. Abortions attributable to naturally acquired N caninum infection also have been observed in pygmy goats. We studied experimentally induced infections with N caninum in pregnant pygmy does to determine whether abortions attributable to N caninum infection would occur after inoculation. Seven pregnant pygmy does (1 control doe and 6 inoculated with N caninum) were studied. The control doe remained clinically normal throughout the study and delivered 2 healthy kids. Abortion, fetal death, and stillbirths were observed in some pregnant does inoculated with N caninum. Two pregnant pygmy does inoculated with N caninum early in gestation (day 51) had fetuses that died and were aborted, or died and were reabsorbed. Neospora caninum tachyzoites and lesions were observed in the brain, spinal cord, and heart of aborted fetuses; parasites also were isolated from the placenta. Four additional pregnant pygmy does (2 inoculated at mid-gestation [day 85], and 2 at late gestation [day 127]) did not abort after inoculation. However, 1 doe inoculated during midgestation delivered a stillborn fetus that had died about 1 week prior to parturition. This kid was congenitally infected with N caninum. Neospora caninum was isolated from the placentas of all inoculated does examined. Neonatal neosporosis was not observed in live-born kids, nor were stages of N caninum isolated from any live-born kid. Does did not undergo abortion or have congenitally infected kids when they were rebred and evaluated for neosporosis.

Free access
in American Journal of Veterinary Research

Summary

Ultrastructure of the interactions of host cell mitochondria with developing Toxoplasma gondii tissue cysts was examined in cultured cells, using transmission electron microscopy of infected cells and rhodamine 123 (a mitochondria-specific vital fluorescent dye) staining of isolated tissue cysts. Structurally mature T gondii tissue cysts were observed as early as 2 days after inoculation of cultured cells. During development of T gondii, host cell mitochondria were observed surrounding the parasitophorous vacuole membrane. Mitochondria became flat and elongated in the vicinity of the parasitophorous vacuole membrane. These mitochondria were also closely associated with T gondii tissue cysts. Incubation of tissue cysts from cultured cells and tissue cysts from mouse brains with rhodamine 123 revealed fluorescence of the tissue cyst wall in living specimens. Incubation of tissue cysts with 10 µM rotenone caused diminished fluorescence of the tissue cyst walls, and 100 µM rotenone caused complete inhibition. Mouse RBC, and tissue cysts fixed in 100% methanol did not fluoresce after exposure to rhodamine. Tissue cysts in 9 isolates of T gondii from mouse brains were examined, using rhodamine 123, and the tissue cyst walls of all isolates fluoresced, indicating no isolate effects. Our results indicate that host cell mitochondria may be closely associated with the tissue cysts of T gondii in cell cultures and in mice.

Free access
in American Journal of Veterinary Research
in Journal of the American Veterinary Medical Association

Abstract

Objective

To test the hypothesis that cats are definitive hosts of Neospora caninum.

Animals

6 weaned male kittens obtained from 2 sources, and several dozen outbred mice.

Procedure

Cats were fed large numbers of 3 strains of N caninum: tissue cysts in buffered saline solution, mouse brain homogenates, and whole carcass homogenates from seropositive mice. Fecal specimens were examined for 4 weeks by use of flotation tests, and bioassays were performed in mice. One cat was inoculated parenterally with tachyzoites, to determine whether cats could respond serologically to N caninum. Tissue cysts from portions of oral inocula were cultured to verify viability. Indirect fluorescent antibody serologic testing, histologic and immunohistologic examinations, cell culture, and polymerase chain reaction procedures were performed 4 to 8 weeks after oral exposure, to seek evidence of infection of cats and mice.

Results

None of the cats or mice seroconverted to N caninum, with the exception of the single cat inoculated parenterally. Fecal shedding of oocysts was not observed, except for Isospora felis oocysts that were shed by 2 cats beginning prior to oral challenge exposure. Evidence of infection was not detected in tissues of cats or mice, with the exception of the parenterally inoculated cat.

Conclusions

The hypothesis that cats are definitive hosts of N caninum is not supported.

Clinical Relevance

Extermination of cats in efforts to control bovine neosporosis is not warranted. (Am J Vet Res 1998;59:441–444)

Free access
in American Journal of Veterinary Research

SUMMARY

The ultrastructure of tachyzoites of 3 isolates of Neospora caninum from dogs was examined, using transmission electron microscopy of infected cultured cells. Ultrastructure of the 3 isolates was similar. Tachyzoites had a pellicle, 22 subpellicular microtubules, a conoid, anterior and posterior polar rings, 8 to 12 electron-dense rhoptries, numerous micronemes, a single vesicular nucleus, tubular mitochondria, Golgi complexes, ribosomes, endoplasmic reticula, an inactive micropore, electron-dense bodies, lipid bodies, and amylopectin bodies. Most tachyzoites were located adjacent to the host cell nucleus in a parasitophorous vacuole that contained numerous intravacuolar tubules. Tachyzoites divided by endodyogeny. Neospora caninum tissue cysts were not seen. Comparison of N caninum with Toxoplasma gondii tachyzoites indicated that the 2 species can be differentiated on the basis of structure and numbers of rhoptries and numbers and location of micronemes and electron-dense bodies.

Free access
in American Journal of Veterinary Research