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At least 217 records · Page 12Linked to original sources
PubMed1991

[The clinical diagnosis of protozoal myositis syndrome (Neospora caninum) of puppies].

Nine puppies suffered from progressive paresis with muscle wasting, hyporeflexia and extensor rigidity. CK-activity in serum was elevated and electrodiagnostic findings were indicative of lower motor neuron disease. Although lesions were also found in the CNS, additional neurological signs were rare, but CSF examination revealed the presence of inflammatory lesions. On pathologic examination, all animals had a disseminated necrotizing myositis. In addition, a disseminated encephalomyelitis was found as well as, in 2 cases, a neuritis. In the lesions of 6 animals protozoal organisms were found which were immunocytochemically identified as Neospora caninum. Our results show that the protozoal myositis-encephalitis syndrome in puppies can be diagnosed in the clinic with high probability. A clinical differentiation between toxoplasmosis and Neospora caninum infection is presently difficult.

Animals↗
PubMed1991

Neospora-like protozoan infection as a major cause of abortion in California dairy cattle.

Ninety-five aborted bovine fetuses received from California dairies over a 4.5-year period had histologic lesions of focal encephalitis. Protozoa that reacted with Neospora caninum antiserum were detected in the brain of 88 of these fetuses and in the heart of 1 fetus. Sarcocystis spp schizonts were seen in the vascular endothelium of 1 fetus. It was concluded that a Neospora-like cyst-forming coccidian may be a major cause of abortion in California dairy cattle.

Abortion, Veterinary↗
PubMed1990

Fatal congenital Neospora caninum infection in a lamb.

Neospora caninum tissue cysts were found in the brain and spinal cord of a 1-wk-old lamb that was unable to stand after birth. The lamb was originally diagnosed as having toxoplasmosis, but ultrastructural and immunohistochemical techniques used in the present study permitted a definitive diagnosis of Neospora caninum tissue cysts in the brain and spinal cord of this lamb. This is the first report of N. caninum in sheep.

Animals↗
PubMed1989

Transplacental Neospora caninum infection in dogs.

A Beagle bitch was inoculated SC and IM with 1.5 million Neospora caninum tachyzoites on the 35th day of gestation. Eight pups were born alive 28 days after N caninum inoculation of the bitch. Pup 1 did not breathe and died while still enclosed in fetal membranes. Pup 2 died 2 days after birth (DAB). Pups 3, 4, and 5 were euthanatized on 2, 3, and 20 DAB, respectively, because they were hypothermic and not nursing. Pups 6, 7, and 8 remained clinically normal. Indirect fluorescent N caninum serum antibody titers were: less than 50 (pups 1 and 8 at 17 DAB, and the bitch before inoculation), 50 (pups 2 and 3 on 2 DAB), 200 (pups 6 and 7 on 17 DAB), and 800 (bitch on day 17 after parturition). Neospora caninum was recovered in cultured cells inoculated with placenta and tissues from all 5 pups necropsied, and N caninum was seen in histologic sections of the heart of pup 5. Results indicate that N caninum can be transplacentally transmitted in dogs.

Animals↗
PubMed1988

Neonatal Neospora caninum infection in dogs: isolation of the causative agent and experimental transmission.

Neospora caninum infection was diagnosed in 5 young dogs from 2 litters with a common parentage. The pups were born healthy, but developed hind limb paresis 5 to 8 weeks after birth. The predominant lesions were polyradiculoneuritis and granulomatous polymyositis. Neospora caninum was seen microscopically in sections of naturally infected pups, and was isolated in cell cultures, mice, and dogs inoculated with infected canine tissues. Antibodies to N caninum were detected in sera of infected dogs by indirect fluorescent antibody test.

Animals↗
PubMed1993

Use of infected cultured cells to compare ultrastructural features of Neospora caninum from dogs and Toxoplasma gondii.

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 fo rhoptries and numbers and location of micronemes and electron-dense bodies.

Animals↗
PubMed2003

Establishment of a two-dimensional electrophoresis map for Neospora caninum tachyzoites by proteomics.

Expressed proteins and antigens from Neospora caninum tachyzoites were studied by two-dimensional gel electrophoresis and immunoblot analysis combined with matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Thirty-one spots corresponding to 20 different proteins were identified from N. caninum tachyzoites by peptide mass fingerprinting. Six proteins were identified from a N. caninum database (NTPase, 14-3-3 protein homologue, NcMIC1, NCDG1, NcGRA1 and NcGRA2), and 11 proteins were identified in closely related species using the T. gondii database (HSP70, HSP60, pyruvate kinase, tubulin alpha- and beta-chain, putative protein disulfide isomerase, enolase, actin, fructose-1,6-bisphosphatase, lactate dehydrogenase and glyceradehyde-3-phosphate dehydrogenase). One hundred and two antigen spots were observed using pH 4-7 IPG strips on immunoblot profiles. Among them, 17 spots corresponding to 11 antigenic proteins were identified from a N. caninum protein map. This study involved the construction of in-depth protein maps for N. caninum tachyzoites, which will be of value for studies of its pathogenesis, drug and vaccine development, and phylogenetic studies.

Animals↗
PubMed2004

Identification of antigenic proteins from Neospora caninum recognized by bovine immunoglobulins M, E, A and G using immunoproteomics.

Antigenic proteins of Neospora caninum (N. caninum) against bovine immunoglobulins M, E, A, and G were investigated by using immunoproteomics. Proteins of N. caninum (KBA-2) tachyzoite lysates separated by two-dimensional gel electrophoresis were transferred to polyvinylidene difluoride (PVDF) membranes, probed with different bovine immunoglobulin class and classified. Antigenic spots recognized were also identified by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) analysis. 132, 84, 4, and 40 antigenic protein spots were recognized on N. caninum immunoblot profiles against bovine IgM, IgE, IgA, and IgG, respectively. Of these protein spots, the antigenic proteins recognized by either IgM, IgE, and IgG, or IgM and IgG were HSP70, pyruvate kinase, actin, NCDG-1, tubulin alpha-chain, and putative ribosomal protein S2. On the other hand, IgM, IgE, and IgA reacted with NTPase, HSP60, tubulin beta-chain, putative protein disulfide isomerase, enolase, lactate dehydrogenase, serine-threonine phosphatase, 14-3-3 protein homologue, and GRA2 protein. Most of the antigenic proteins identified were associated with the process of invasion, proliferation, and egression of apicomplexans. In our study, HSP70, actin, NTPase, HSP60, pyruvate kinase, enolase, putative ribosomal protein S2, NCDG-1, and GRA2 proteins were found to be immunodominant proteins, which may contribute to the development of diagnostic markers and vaccine.

Animals↗
PubMed1997

Neospora caninum: role for immune cytokines in host immunity.

Neospora caninum is a coccidial protozoan parasite that infects a large range of mammals including dogs, cats, mice, and cattle. Morphologically, N. caninum appears indistinguishable from Toxoplasma gondii, although they are genetically distinct. To date there have been no reported cases of this infection in humans, although nonhuman primates may be susceptible to infection. Inbred A/J mice develop no clinical and little histologic evidence of infection in spite of a high-dose inoculum of N. caninum. Splenocytes obtained from infected mice proliferate in vitro in response to both N. caninum and T. gondii-soluble antigen. A transient state of T cell hyporesponsiveness to parasite antigen and mitogen was observed at Day 7 p.i. This downregulatory response could be partially reversed by the addition of the nitric oxide antagonist LNMMA, but not antibody to IL-10. Mice infected with N. caninum produce significant quantities of IL-12 and IFN gamma, most evident shortly after infection. In vivo, antibody to IL-12 is able to neutralize immune resistance to the parasite. Moreover, in vivo depletion of IFN gamma with antibody renders the mice susceptible to infection. These observations suggest that N. caninum induces a T cell immune response in the infected host that is at least partially mediated by IL-12 and IFN gamma.

Animals↗
PubMed1998

Neospora caninum: tachyzoites express a potent type-I nucleoside triphosphate hydrolase.

We have identified type I nucleoside triphosphate hydrolase (NTPase; EC 3.6. 1.3) activity, previously thought to be restricted to the virulent strains of Toxoplasma gondii, in the cell extracts of Neospora caninum tachyzoites. Sequence analysis of a complete cDNA from Nc-1 strain indicated that N. caninum NTPases shared approximately 69% identity to the NTPases of T. gondii and are most similar to the NTPase-I isozyme. Southern blot analysis of genomic DNA and sequence analysis of two independent NTP clones from the Nc-1 strain revealed the presence of multiple genes, at least two of which are transcribed. Substrate specificity and Km values for MgATP2- and MgADP- hydrolysis for recombinant or partially purified native NcNTPase were the same as those for the type I isozyme (NTPase-I). Significantly, no type II enzyme (NTPase-II) activity for NDP hydrolysis was detected in cell extracts of N. caninum, although it is universally present in all T. gondii strains that have been tested. This intriguing difference between these two closely related apicomplexan parasites may provide insight into the function of the NTPases during intracellular parasitism.

Acid Anhydride Hydrolases↗
PubMed1999

Neospora caninum: identification of 19-, 38-, and 40-kDa surface antigens and a 33-kDa dense granule antigen using monoclonal antibodies.

Neospora caninum, a coccidian parasite closely related to Toxoplasma gondii, can infect a broad host range and is regarded as an important cause of bovine abortion worldwide. In the present study, four antigens of N. caninum were partially characterized using monoclonal antibodies. Immunofluorescence of viable tachyzoites as well as the immunoprecipitation of antigens extracted from tachyzoites previously labeled by surface biotinylation revealed that three of these antigens with apparent molecular weights of 40, 38, and 19 kDa are located in the outer surface membrane of this parasite stage. Further evidence for the surface localization of the 38-kDa antigen was obtained by immunoelectron microscopy. In addition to the surface molecules, an antigen located in dense granules and in the tubular network of the parasitophorous vacuole was detected by another monoclonal antibody. When tachyzoite antigens separated under nonreducing conditions were probed on Western blots, this antibody reacted mainly with a 33-kDa antigen. Immunohistochemical analysis of infected tissue sections indicated that the 33-kDa dense granule antigen is present in both tachyzoites and bradyzoites, while the 38-kDa surface antigen from tachyzoites seems to be absent in bradyzoites.

Animals↗
PubMed1996

Discrimination of Neospora caninum from Toxoplasma gondii and other apicomplexan parasites by hybridization and PCR.

Neospora caninum is a protozoan parasite which causes neurological problems in dogs and abortion in cattle. As N. caninum is difficult to distinguish morphologically from Toxoplasma gondii, we developed a molecular tool capable of discriminating between the two parasites. Genomic DNA was isolated from in vitro cultured N. caninum tachyzoites and cloned into a plasmid vector. Resulting colonies were subsequently screened by differential hybridization using N. caninum and T. gondii DNA. Two clones were characterized in detail: one clone, termed pNc5, was found to be specific for N. caninum whereas the second clone, pNc1, hybridized with DNA from both parasites. The sequence of pNc5 was determined and different oligonucleotide primers were designed for use in the polymerase chain reaction (PCR). A 944 bp fragment was specifically amplified from N. caninum DNA, but not from DNA extracted from T. gondii or different Sarcocystis species. Positive signals in PCR were obtained with as little as 100 pg parasite template DNA. In addition, dual PCR with primer pairs specific for N. caninum and T. gondii allowed the detection of either parasite in mixed samples.

Animals↗
PubMed2000

Canine neosporosis: clinical and pathological findings and first isolation of Neospora caninum in Germany.

Neosporosis was diagnosed in an 11-week-old puppy of the breed Kleiner Münsterländer with progressive hindlimb paresis. Pathohistological and immunohistological examinations revealed a disseminated infection with Neospora caninum. Parasitic stages were demonstrated in the brain, spinal cord, retina, muscles, thymus, heart, liver, kidney, stomach, adrenal gland, and skin. Immunohistochemistry investigations were carried out using polyclonal rabbit antisera developed against N. caninum tachyzoites and the recombinant bradyzoite-specific antigen BAG-5 of Toxoplasma gondii, which is known to cross-react with N. caninum bradyzoites. BAG-5 antibodies recognized tissue cysts within the CNS and some protozoan stages that were not surrounded by a visible cyst wall. All parasite clusters in the retina and some in muscle tissue stained positively with the BAG-5 antiserum. N. caninum was isolated in cell culture and mice inoculated with brain and spinal cord of the puppy. The new isolate is the first reported in Germany and is designated NC-GER1.

Animals↗
PubMed2000

Neospora caninum: its oocysts and its identity: an opinion.

A recently published opinion that Hammondia heydorni, Neospora caninum, and N. hughesi are a single species is not supported by genetic comparisons. Furthermore, the suggestion that oocysts shed by dogs after experimental infection with N. caninum might have been of another organism is not consistent with the finding that these oocysts induced N. caninum infections in mice.

Animals↗
PubMed2004

Structure analysis of two Toxoplasma gondii and Neospora caninum satellite DNA families and evolution of their common monomeric sequence.

A family of repetitive DNA elements of approximately 350 bp-Sat350-that are members of Toxoplasma gondii satellite DNA was further analyzed. Sequence analysis identified at least three distinct repeat types within this family, called types A, B, and C. B repeats were divided into the subtypes B1 and B2. A search for internal repetitions within this family permitted the identification of conserved regions and the design of PCR primers that amplify almost all these repetitive elements. These primers amplified the expected 350-bp repeats and a novel 680-bp repetitive element (Sat680) related to this family. Two additional tandemly repeated high-order structures corresponding to this satellite DNA family were found by searching the Toxoplasma genome database with these sequences. These studies were confirmed by sequence analysis and identified: (1). an arrangement of AB1CB2 350-bp repeats and (2). an arrangement of two 350-bp-like repeats, resulting in a 680-bp monomer. Sequence comparison and phylogenetic analysis indicated that both high-order structures may have originated from the same ancestral 350-bp repeat. PCR amplification, sequence analysis and Southern blot showed that similar high-order structures were also found in the Toxoplasma-sister taxon Neospora caninum. The Toxoplasma genome database (http://ToxoDB.org ) permitted the assembly of a contig harboring Sat350 elements at one end and a long nonrepetitive DNA sequence flanking this satellite DNA. The region bordering the Sat350 repeats contained two differentially expressed sequence-related regions and interstitial telomeric sequences.

Animals↗
PubMed2004

Effects of toltrazuril and ponazuril on the fine structure and multiplication of tachyzoites of the NC-1 strain of Neospora caninum (a synonym of Hammondia heydorni) in cell cultures.

Rhesus monkey kidney cell cultures were used to propagate tachyzoites of the NC-1 strain of Neospora caninum (syn. Hammondia heydorni). The infected cell cultures were incubated for 4-12 h in media containing 0, 1, 10 or 100 microg/ml of either toltrazuril or ponazuril. The effects were studied by light and electron microscopy. Drug dosages of at least 30 microg/ml were needed to eliminate the parasites. Ponazuril was found (with respect to the reduction of the number of parasites) to be less effective at dosages of 30 microg/ml compared to toltrazuril. However, the damage to the tachyzoites being incubated in 30 microg toltrazuril or ponazuril seen by electron microscopy was so significant that it was surely lethal. The initial damage occurred within the apicoplast and the tubular mitochondrion in all cases,thus destroying two of the most important cell organelles.

Animals↗
PubMed2004

The role of B- and T-cell immunity in toltrazuril-treated C57BL/6 WT, microMT and nude mice experimentally infected with Neospora caninum.

Neospora caninum is a protozoan parasite predominantly known for causing abortion in cattle and neuromuscular disease in dogs. So far, no efficient metaphylactic chemotherapy has been developed. In preliminary studies, toltrazuril had been successfully used against experimental neosporosis in mice and calves. In the present study, we used immunocompetent and immunodeficient mouse strains to address the role of immunity in supporting the chemotherapy of experimental N. caninum infection. WT, microMT and athymic nude mice were intraperitoneally inoculated with 1x10(6) Nc-1 tachyzoites. The drug was administered in the drinking water for 6 consecutive days so as to obtain a daily dose of approximately 20 mg toltrazuril/kg body weight. The course of infection was monitored by clinical, histological and immunohistochemical means, as well as by the search for parasite DNA using PCR-analyses of various organs. In immunocompetent WT mice, treatment proved to be of high efficacy by abrogation of any lesion formation or PCR-positivity in medicated C57BL/6 mice and a significant reduction of lesion formation or PCR-positivity in BALB/c animals. Similarly, treated microMT mice exhibited a significant reduction in cerebral lesion formation as well as in parasite DNA detectability by PCR when compared to untreated animals. Athymic nude mice, however, did not respond to treatment in that only a delay of the parasite dissemination was achieved, and nude mice still showed the neosporosis disease symptoms, although later than untreated animals. We conclude that treatment with toltrazuril appears to act parasitostatically rather than parasitocidically. This is supported by the fact that: (1) although the lack of B-cells did not impair the effect of toltrazuril, (2) the lack of T-cells did not allow for a full efficacy of treatment. Therefore, chemotherapy with toltrazuril against experimental infections with N. caninum requires the support of T-cell immunity in order to be successful.

Administration, Oral↗