Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Neospora”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 505 records · Page 28Linked to original sources
PubMed2002

Coprodiagnosis of Hammondia heydorni in dogs by PCR based amplification of ITS 1 rRNA: differentiation from morphologically indistinguishable oocysts of Neospora caninum.

Hammondia heydorni is thought to be a non-pathogenic coccidian parasite of dogs that is closely related to Neospora caninum, an important parasite of cattle and dogs. Oocysts of these two species are morphologically indistinguishable from each other. A population of 2240 dogs in the Czech Republic was screened for the presence of H. heydorni/N. caninum oocysts and five (0.22%), represented by five of 3135 faecal samples (0.16%), were positive. The internal transcribed spacer 1 region of the rRNA gene (ITS1) from two isolates were cloned and the DNA sequences were identical with those of the ITS1 of H. heydorni. Based on the rRNA sequences available for H. heydorni and related coccidia, the primer pair JS4-JS5 was designed to amplify the 3' end of the small subunit (SSU) rRNA gene and ITS1 of H. heydorni. When tested on DNA extracted from a variety of parasites, the primers amplified a specific 267 bp fragment in our isolates only. The presence of DNA equivalent to 10 oocysts was sufficient for the amplification of the ITS1. We present a PCR-based diagnostic method as the only fast and reliable method for the diagnosis of H. heydorni in dogs.

Animals↗
PubMed2005

Associations between pregnancy outcome and serological response to Neospora caninum among a group of dairy heifers.

AIM: To monitor pregnancy in a group of rising 2-year-old dairy heifers on a farm on which abortion due to Neospora caninum was known to occur in previous years. METHODS: A prospective cohort study group of 164 rising 2-year-old heifers was pregnancy-tested and blood-sampled at 4-5-week intervals throughout gestation. Sera were tested for antibodies to N. caninum at 3-4-month intervals, using an enzyme-linked immunosorbent assay (ELISA). When loss of pregnancy was detected, an N. caninum indirect fluorescent antibody test (IFAT) was conducted retrospectively on stored sera collected the month before abortion, the month abortion was detected, and for the following 2 months, from heifers that aborted. All fetal and placental material detected following abortion was subjected to gross post-mortem and histopathological examination. RESULTS: Eleven of 18 (61%) heifers that were seropositive and 4/146 (3%) heifers that were seronegative to N. caninum by ELISA, aborted. The relative risk for abortion among ELISA-positive heifers was 23.6. Abortion occurred predominantly between Days 120 and 152 gestation among the ELISA-positive heifers and throughout gestation among the ELISA-negative heifers. IFAT titres rose around the time of abortion in most of the heifers that were previously seropositive by ELISA, but dropped rapidly again in post-abortion samples. IFAT titres among 4/6 ELISA-positive heifers that did not abort increased, but later in gestation than the time other heifers aborted. IFAT titres remained negative in heifers that aborted that were ELISA negative. CONCLUSIONS: Heifers that were seropositive to N. caninum by ELISA had a much greater risk of abortion than seronegative heifers. Most seropositive heifers showed evidence of a reactivation of infection during pregnancy. High (> or =1:2,000) N. caninum IFAT titres also occurred in non-aborting heifers. CLINICAL RELEVANCE: Culling of replacement heifers seropositive to N. caninum may be a cost-effective strategy for minimising risk of abortion. Pregnancy testing heifers before 5 months gestation may overestimate the number that calve in N. caninum-infected herds, but would assist in documenting the occurrence of abortion. Reliance on a high (>1:2,000) IFAT titre to rule-in N. caninum as a cause of abortion is likely to produce false-positive results.

Abortion, Veterinary↗
PubMed2004

Detection of the protozoan Neospora caninum using in situ polymerase chain reaction.

A new method to detect the protozoan Neospora caninum using indirect in situ polymerase chain reaction (PCR) is described. In situ PCR combines the advantages of the extraordinarily high sensitivity and specificity of PCR and the in situ representation of immunohistochemical methods. We describe an indirect in situ PCR, whereby the amplified products were detected using a primed in situ (PRINS) reaction with hapten-labeled nucleotides and visualized using fluorochrome-labeled antibodies. This technique was carried out in both infected cell cultures and formalin fixed, paraffin embedded tissues. Clear signals were obtained in the N. caninum positive samples using in situ PCR, whereas control slides with Toxoplasma gondii infected tissues always yielded negative results.

Animals↗
PubMed2004

Recognition patterns of Neospora caninum tachyzoite antigens by bovine IgG at different IFAT titres.

This study describes qualitative and quantitative antibody response in cows naturally infected with Neospora caninum. The study was carried out with 269 serum samples obtained from 24 cows over a period of 15 weeks. Prior to sample collection, the cows were tested with ELISA. The 269 samples were screened with IFAT and categorized into seven IFAT titre groups (< 1 : 80, 1 : 80, 1 : 200, 1 : 600, 1 : 1000, 1 : 2000, > 1 : 2000). The samples were finally analysed by Western blotting. Seven immunodominant antigens (approximately 18-, approximately 25-, approximately 33-, approximately 35-36-, approximately 45-46-, approximately 47-, approximately 60-62 kDa) and five minor antigens (approximately 25, approximately 51, approximately 64, approximately 77, approximately 116 kDa) were recognized by cow sera. The recognition of approximately 46 kDa antigen by cow sera was common to samples with IFAT titre 1 : 80 and above. Another common antigen was the approximately 18 kDa antigen, which was recognized by samples with IFAT titre 1 : 200 and above. The most remarkable observation was the presence of the 45-46 kDa, the 77 kDa, and absence of the 18 kDa antigenic bands in samples with IFAT titre 1 : 80. This observation was consistent even in the face of fluctuating antibody titre where serum antibody titres from an animal exceeded then failed to reach 1 : 80. Antibody fluctuation was observed across all cows (pregnant and aborted) with no discernible fluctuation pattern. However, the fluctuation in antibody titre observed appeared to be most remarkable in initially ELISA-negative pregnant cows, and to a lesser extent in ELISA-positive pregnant cows, and ELISA-positive aborted cows. Although there was fluctuation in antibody titre, the banding patterns of N. caninum tachyzoite antigens by cows within the same IFAT titre group remained similar.

Abortion, Veterinary↗
PubMed2004

Establishment of Neospora caninum antigen-specific T cell lines of primarily CD4 T cells.

Neosporosis is an important cause of pregnancy loss in cattle worldwide. Protective immunity against Neospora caninum infection may include both cell-mediated (CMI) and humoral immune responses. This study was to establish short-term antigen-specific T cell lines composed of primarily CD4(+)T cells from peripheral blood lymphocytes (PBL) of infected cows, which may be used to identify immunodominant antigens for the development of N. caninum vaccines. Crude N. caninum tachyzoite antigen was prepared from in vitro derived N. caninum tachyzoites. Multiple T cell lines were established and maintained for 11 weeks by weekly re-stimulation with N. caninum antigen and antigen-presenting cells. All cell lines responded highly to antigen between weeks 3 and 11. Phenotypically, these cells were composed primarily of CD4(+)T cells between weeks 2-8, with a gradual expansion of gamma/delta(+)T cells thereafter. The results indicate that N. caninum-specific T cell lines can be established and maintained without exogenous T cell growth factors and may be used to identify N. caninum antigens. This research will enhance our understanding of bovine CMI to neosporosis and may facilitate development of a proven neosporosis vaccine.

Animals↗
PubMed2005

Characterization of the B-cell immune response elicited in BALB/c mice challenged with Neospora caninum tachyzoites.

Activation of B cells occurring in hosts infected with protozoan parasites has been implicated either in protective or parasite-evasion immune-mediated mechanisms. Intraperitoneal inoculation of Neospora caninum tachyzoites into BALB/c mice induces an acute response characterized by a rapid increase in the numbers of CD69-expressing peritoneal and splenic B cells. This early B-cell stimulatory effect preceded an increase in the numbers of total and immunoglobulin-secreting splenic B cells and a rise in serum levels of N. caninum-specific immunoglobulins, predominantly of the immunoglobulin G2a (IgG2a) and IgM isotypes. Increased numbers of B cells expressing the costimulatory molecules CD80 and CD86 were also observed in the N. caninum-infected mice. The B-cell stimulatory effect observed in mice challenged with N. caninum tachyzoites was reduced in mice challenged with gamma-irradiated parasites. Contrasting with the peripheral B-cell expansion, a depletion of B-lineage cells was observed in the bone-marrow of the N. caninum-infected mice. Intradermal immunization of BALB/c mice with diverse N. caninum antigenic preparations although inducing the production of parasite-specific antibodies nevertheless impaired interferon-gamma (IFN-gamma) mRNA expression and caused lethal susceptibility to infection in mice inoculated with a non-lethal parasitic inoculum. This increased susceptibility to N. caninum was not observed in naïve mice passively transferred with anti-N. caninum antibodies. Taken together, these results show that N. caninum induces in BALB/c mice a parasite-specific, non-polyclonal, B-cell response, reinforce previous observations made by others showing that immunization with N. caninum whole structural antigens increases susceptibility to murine neosporosis and further stress the role of IFN-gamma in the host protective immune mechanisms against this parasite.

Animals↗
PubMed1994

Neospora caninum in dogs: detection of antibodies by ELISA using an iscom antigen.

An indirect enzyme linked immunsorbent assay (ELISA) for detection of antibodies to Neospora caninum in serum from dogs is described. Extracted tachyzoite proteins incorporated into immunostimulating complexes (iscoms) were used as coating antigen. A mixture of a monoclonal antibody to dog immunoglobulin G and a horse radish peroxidase conjugated antibody to mouse Ig was used to detect bound antibody. When the iscom preparation was analysed by means of sodium dodecyl sulphate polyacrylamide gel electrophoresis it appeared to consist of a restricted number of proteins compared with whole parasite homogenates. In immunoblot analysis, using N. caninum positive sera from rabbits and dogs as probes, the major antigens recognized had approximate molecular weights between 30 and 45 and 17 to 19 kDa. Compared with an ELISA using a crude solubilized tachyzoite antigen, the iscom ELISA substantially improved the sensitivity and specificity (to 97.6% and 95.6%, respectively, against an immunofluorescence test, IFAT, as indicator of true status). There was a statistically significant positive correlation between IFAT titres and iscom ELISA OD450 values. The iscom ELISA absorbances (and the IFAT titres) of dogs with proven clinical infections were not higher than those from nonclinically affected, putatively infected dogs.

Animals↗
PubMed2007

Analysis of the immune response to Neospora caninum in a model of intragastric infection in mice.

To study experimental Neospora caninum infection initiated at the gastrointestinal tract, Toll-like Receptor 4- and functional IL-12Rbeta2 chain-deficient C57BL/10 ScCr mice were challenged intragastrically with 5 x 10(6) N. caninum tachyzoites. All parasite-inoculated mice eventually died with disseminated infection. In contrast, immunocompetent BALB/c mice challenged with 1 x 10(7) N. caninum tachyzoites by the intragastric (i.g.) or the intraperitoneal (i.p.) route remained alive for at least 6 months. Expansion of splenic B- and T-cells, the latter displaying both activated and regulatory phenotypes, and increased levels of IFN-gamma and IL-10 mRNA were detected in both groups of infected BALB/c mice compared with non-infected controls, whereas in the Peyer's patches only IFN-gamma mRNA levels were found to be increased. Parasite-specific IgG1, IgG2a and IgA antibody levels were elevated in the sera of all infected mice, whereas increased N. caninum-specific IgA levels were detected in intestinal lavage fluids of i.g. challenged mice only. These results show that N. caninum infection can be successfully established in mice by i.g. administration of tachyzoites. They also show that the immune response elicited in i.g. or i.p. infected BALB/c mice, although conferring some degree of protection, was not sufficient for complete parasite clearance.

Animals↗
PubMed2004

Meningoencephalomyelitis caused by Neospora caninum in a juvenile fallow deer (Dama dama).

Neosporosis, caused by the protozoan parasite Neospora caninum, is a serious cause of bovine abortion, stillbirth and perinatal death. This paper reports a clinical neosporosis in a 3-week-old fallow deer (Dama dama). The fawn was full term and appeared normal at birth. Histological lesions consisted of a multifocal necrotizing and granulomatous meningoencephalomyelitis, with intralesional protozoal cysts. Positive immunohistochemical staining and characteristic PCR products confirmed the diagnosis of N. caninum infection.

Animals↗
PubMed2005

Adaptation of a surface antigen-based ELISA for the detection of antibodies against Neospora caninum in bovine milk.

The aim of the present study was to determine whether an ELISA based on a 38-kDa surface antigen (NcSRS2) of Neospora caninum tachyzoites could be used to examine bovine milk for antibodies against N. caninum. A total of 797 undiluted milk samples from N. caninum-infected herds were examined in the p38-ELISA. Milk results of individual animals were compared with those obtained by the same ELISA for the corresponding serum samples. The linear correlation between milk and serum antibody results of individual animals was characterized by an adjusted R(2) of 0.644. The examination of the two-graph receiver-operating characteristics revealed an optimal cut-off of 0.150 to obtain similar results in the examination of milk and serum. With this cut-off, the test had a specificity and a sensitivity relative to the serum results of 93%. Using this cut-off, excellent agreement was observed between milk and serum results (Kappa 0.85; 95% CI, 0.781-0.920). A cut-off of 0.450 resulted in a specificity of 99% relative to the serum results. At this cut-off, the sensitivity of the test was 80% relative to the serum-ELISA and agreement was slightly lower (Kappa = 0.82; 95%CI, 0.749-0.887). An optimized linear regression model suggests that, in addition to the result in the p38-serum-ELISA, the variables abortion risk (ABRISK) (abortion between 100 and 269 days pregnant) and the age of the animal (AGE) affected the result of the p38-milk-ELISA. The optimized linear regression model had an adjusted R(2) of 0.8501.

Animals↗
PubMed2005

Relationship between rainfall and Neospora caninum-associated abortion in two dairy herds in a dry environment.

The possible direct relationship between climate variations and abortion in Neospora caninum-infected cows has not been studied. The objective of this study was to determine whether climate changes could be a risk factor for abortion in N. caninum-infected cows, and was based on yearly serological screening for neosporosis and on the confirmation of N. caninum infection on aborted fetuses in two high-producing dairy herds with a mean 27% seroprevalence of N. caninum antibodies. The final population study was comprised of 357 pregnancies in seropositive animals. Logistic regression analysis indicated no significant effects of herd, N. caninum antibody titre, climate variables during the first and third trimesters of gestation, mean and maximum temperature-humidity index values during the second trimester of gestation, and previous abortion on the abortion rate. Based on the odds ratio, a 1-unit increase in lactation number yielded a 0.85-fold decrease in the abortion rate. The likelihood of abortion was 1.9 times (1/0.54) lower for pregnant cows inseminated with beef bull semen compared with Holstein-Friesian bull semen. The likelihood of abortion decreased significant and progressively by factors of 0.5, 0.41 and 0.3 for the respective classes 40-49, 30-39 and <30 rainfall mm during the second trimester of gestation (using the class >or=60 rainfall mm as reference). As a general conclusion, it seems that increased rainfall in a dry environment can compromise the success of gestation in N. caninum-infected cows. Attempts should therefore be made to reduce environment effects during the second trimester of gestation, a period in which the immune response of cows is diminished.

Abortion, Veterinary↗
PubMedDate not supplied

Phylogenetic analysis of coccidia based on 18S rDNA sequence comparison indicates that Isospora is most closely related to Toxoplasma and Neospora.

The phylogenetic relationships and taxonomic affinities of coccidia with isosporan-type oocysts have been unclear as overlapping characters, recently discovered life cycle features, and even recently discovered taxa, continue to be incorporated into biological classifications of the group. We determined the full or partial 18S ribosomal RNA gene sequences of three mammalian Isospora spp., Isospora felis, Isospora ohioensis and Isospora suis, and a Sarcocystis sp. of a rattlesnake, and used these sequences for a phylogenetic analysis of the genus Isospora and the cyst-forming coccidia. Various alveolate 18S rDNA sequences were aligned and analyzed using maximum parsimony to obtain a phylogenetic hypothesis for the group. The three Isospora spp. were found to be most closely related to Toxoplasma gondii and Neospora caninum. This clade in turn formed the sister group to the Sarcocystis spp. included in the analysis. The results confirm that the genus Isospora does not belong to the family Eimeriidae, but should be classified together with the cyst-forming coccidia in the family Sarcocystidae. Furthermore, there appear to be two lineages within the Sarcocystidae. One lineage comprises Isospora and the Toxoplasma/Neospora clade which share the characters of having a proliferative phase of development preceding gamogony in the definitive host and an exogenous phase of sporogony. The other lineage comprises the Sarcocystis spp. which have no proliferative phase in the definitive host and an endogenous phase of sporogony.

Animals↗
PubMedDate not supplied

Vaccination with gamma-irradiated Neospora caninum tachyzoites protects mice against acute challenge with N. caninum.

Neospora caninum, an apicomplexan parasite, is a leading cause of bovine abortions worldwide. The efficacy of gamma-irradiated N. caninum strain NC-1 tachyzoites as a vaccine for neosporosis was assessed in C57BL6 mice. A dose of 528 Gy of gamma irradiation was sufficient to arrest replication but not host cell penetration by tachyzoites. Female C57BL6 mice were vaccinated with two intraperitoneal inoculations of 1 x 10(6) irradiated tachyzoites at 4-wk intervals. When stimulated with N. caninum tachyzoite lysates, splenocytes of vaccinated mice, cultured 5 and 10 wk after vaccination, secreted significant (P<0.05) levels of interferon gamma, interleukin (IL)-10, and small amounts of IL-4. Antibody isotype-specific ELISA of sera from vaccinated mice exhibited both IgG1 and IgG2a isotypes of antibodies. Vaccinated mice were challenged intraperitoneally with 2 x 10(7)N. caninum tachyzoites. All vaccinated mice remained healthy and showed no obvious signs of neosporosis up to the 25th day post-challenge when the study was terminated. All unvaccinated control mice died within 1 wk of infection. Gamma-irradiated N. caninum tachyzoites can serve as an effective, attenuated vaccine for N. caninum.

Animals↗
PubMed1995

Neospora caninum infection in three dogs.

Infection with Neospora caninum in three young dogs is described. The predominant clinical signs were lower motor neuron deficits of the pelvic limbs, bladder and rectum. In two cases there was liver infection and dysfunction. The younger dogs had an acute onset rapidly progressive syndrome. The older dog had a similar but more chronic course. The diagnosis was confirmed by an immunofluorescence antibody test. The parasite is sensitive to clindamycin and trimethoprim/sulphonamide preparations, however the prognosis for return to function is poor especially if muscle contracture has occurred.

Acute Disease↗
PubMed1996

Distribution of Neospora caninum within the central nervous system and other tissues of six dogs with clinical neosporosis.

Six cases of neosporosis in dogs were diagnosed on the basis of clinical signs, serology and immunohistochemistry. The brains and spinal cords, at several levels, and various other tissues were examined using anti-Neospora caninum serum by an immunoperoxidase technique. Parasite density varied markedly from case to case. Although found most consistently in the cerebrum, parasites were distributed throughout the central nervous system (CNS), both within the grey and white matter and within nerve roots. Clinical signs were not related to the position of parasites. Tissue cysts were found infrequently in all areas of the CNS, but not in other tissues. Parasite density was not related to the age of the dog or whether treatment had been given. However, with the exception of the only adult dog examined, more CNS parasites were found in dogs with a longer duration of illness and with higher antibody titres. Tachyzoites were present in skeletal muscles, in the muscularis of the oesophagus, in heart, lung and, less frequently, liver, and rarely in the adrenal gland, thyroid gland and uterus; no clinical signs were seen resulting from damage to these organs. Parasites were not observed in lymphoid tissue. In visceral organs, parasites were most widely distributed in peracute cases. For post mortem diagnosis of neosporosis, the CNS, particularly the cerebrum, is the optimum tissue to examine but parasites may also be found in many other tissues, especially in acute cases. Muscle biopsy of appropriate muscles (as suggested by the clinical signs) provides the possibility of a definitive premortem diagnosis.

Animals↗
PubMed1997

Seroprevalence of antibodies to Neospora caninum in Belgian dogs.

Sera from 300 dogs from Ghent and Antwerp were tested for antibodies to Neospora caninum using an indirect fluorescent antibody test. Overall, 11 per cent (9.5 to 13 per cent; confidence interval of 95 per cent) of dogs were seropositive, at titres of 1:50 to 1:800. No sex or breed differences were detected, but there was an increase in seropositivity with age.

Aging↗
PubMed2000

Horner's syndrome associated with Neospora infection.

A working collie cross was presented with a three-month history of vague neurological signs and a right-sided Horner's syndrome. Denervation hypersensitivity testing suggested a first order syndrome. There was a significant positive titre to Neospora and clinical signs resolved completely following treatment.

Animals↗
PubMed2000

Seroprevalence of Neospora caninum infection following an abortion outbreak in a dairy cattle herd.

OBJECTIVE: To investigate the seroprevalence of Neospora caninum infection in a commercial dairy cattle herd, 15 months after detection of an abortion outbreak. PROCEDURE: Sera from the whole herd (n = 266) were examined for N caninum antibodies by indirect fluorescent antibody test (IFAT) and immunoblot analysis. Herd records were reviewed to collate serological results with abortion history, proximity to calving, and pedigree data. RESULTS: The seroprevalence of N caninum infection was 24% (63/266) for IFAT titre > or = 160, 29% (78/266) for immunoblot positive (+ve), and 31% (82/266) for IFAT > or = 160 and/or immunoblot +ve; 94% (59/63) of animals with IFAT > or = 160 were immunoblot +ve. The association between seropositivity (IFAT > or = 160 and/or immunoblot +ve) and history of abortion was highly significant (P < 0.001); the seroprevalence was 86% (18/21) in aborting cows, compared with 30% (50/164) in non-aborting animals. The abortion rate for seropositive cows was 26% (18/68) compared with 3% (3/117) for seronegative animals. IFAT titres of infected cows were higher within 2 months of calving than at other times (P < 0.001). The association between seropositivity in dams and daughters was highly significant (P = 0.009). CONCLUSIONS: The abortions were associated with N caninum infection and there was evidence of reactivation of latent infection close to calving and congenital transmission of infection. Immunodominant antigens identified by immunoblots may prove useful for improved diagnostic tests.

Abortion, Veterinary↗