PMID(sorted ascending)
development of neospora caninum cultured with human serum in vitro and in vivo.because there has been no report of symptomatic neospora caninum infection in humans, we examined the effect of human serum on the parasite's growth in either a bovine angioendothelial cell or caco-2 cell culture in vitro and in immunocompromised mice in vivo. there was no difference in intracellular parasite numbers between cells incubated with human serum at 24 hr after challenge and those incubated with fetal bovine serum (fbs), which has no titer for the anti-n. caninum agglutination antibod ...200515856911
serological survey of selected pathogens of free-ranging foxes in southern argentina, 1998--2001.a serological survey was conducted in the patagonia region of argentina to estimate the prevalence of nine disease agents within the populations of free-ranging culpeo (dusicyon culpaeus) and grey (dusicyon griseus) foxes. the disease agents were aujeszky's disease virus (adv), brucella, canine adenovirus (cav), canine distemper virus (cdv), canine parvovirus (cpv), encephalitozoon cuniculi, leptospira, neospora caninum, and toxoplasma gondii. a total of 84 foxes were sampled (28 culpeo and 56 g ...200415861875
cutaneous manifestations of disseminated toxoplasmosis in an immunosuppressed dog.a 3.5-year-old, castrated male, giant schnauzer was presented with alopecic pustular dermatitis. immune-mediated hemolytic anemia had been diagnosed 45 days previously. at the time of presentation, the dog was receiving prednisone, azathioprine, and cyclosporine. cutaneous protozoosis was diagnosed, and postmortem examination revealed protozoa within cutaneous, cardiac, pancreatic, and pulmonary tissues. the protozoa divided by endodyogeny, had the morphology of toxoplasma gondii (t. gondii) tac ...200515870255
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 populat ...200515876229
a review of neospora caninum in dairy and beef cattle--a canadian perspective.neospora caninum is one of the most important causes of abortion in cows. the occurrence of n. caninum infection in beef and dairy cattle has been reported worldwide, and in most provinces in canada. the objective of this review is to summarize our current understanding of n. caninum in dairy and beef cattle for canadian bovine practitioners. the review covers the life cycle of the agent, its mechanisms of transmission, clinical signs, and tests for diagnosing the infection. data on the prevalen ...200515884645
reduction in transplacental transmission of neospora caninum in outbred mice by vaccination.infection with the protozoan parasite neospora caninum is an important cause of abortion in cattle. a major source of infection is transplacental transfer of the parasite from mother to offspring during pregnancy. this study describes investigations on the immunisation of outbred qs mice before pregnancy with live or a crude lysate of n. caninum (nc-nowra isolate) to prevent transplacental transfer of a challenge infection administered during pregnancy. parasites present in the brains of pups fr ...200515885695
seroprevalence of neospora caninum in dairy and beef cattle with reproductive disorders in japan.serum samples from 145 dairy and 65 beef cattle with reproductive disorders and 54 normally calving dairy cattle (controls) in japan were tested for presence of neospora caninum antibodies by use of an indirect fluorescent antibody test (ifat, titer 1:200). overall, seroprevalence of n. caninum was significantly higher (p < 0.001) in dairy cattle (20.0%, 29/145) than in beef cattle (1.5%, 1/65). in cattle which aborted, seroprevalence of n. caninum was significantly higher (p = 0.041) in dairy c ...200515893065
immune response to neospora caninum in naturally infected heifers and heifers vaccinated with inactivated antigen during the second trimester of gestation.the objective of this study was to compare the immune response to neospora caninum in naturally infected heifers and heifers inoculated with a killed whole n. caninum tachyzoite preparation during the second trimester of gestation. nine holstein heifers were used in this study; three naturally infected heifers were born from seropositive dams, and six seronegative heifers were born from seronegative dams. four seronegative heifers were subcutaneously vaccinated with a killed whole n. caninum tac ...200515893067
serologic immunoreactivity to neospora caninum antigens in dogs determined by indirect immunofluorescence, western blotting and dot-elisa.neospora caninum, is a coccidian protozoan known as a major cause of bovine abortion and canine neuropathies. the aim of the present study was to develop a reliable and quick test to detect antibodies to n. caninum in dog sera. sixty-five serum samples from dogs, including 35 positive and 30 negative for n. caninum antibodies were used for standardization of the test. in parallel, immunoreactivity of the sera to toxoplasma gondii antigens was investigated using a passive agglutination test. a do ...200515893072
detection of specific antibodies to neospora caninum and toxoplasma gondii in naturally infected alpacas (lama pacos), llamas (lama glama) and vicunas (lama vicugna) from peru and germany.sera of an experimentally neospora caninum infected llama and a non-infected control llama were used to establish an immunoblot, an elisa and an ifat to detect antibodies against n. caninum tachyzoites. subsequently, serum samples collected from a total of 871 south american camelids (sac: lama glama, lama pacos, lama vicugna) of two farms in peru and from 32 sac of a farm in central germany were examined for antibodies against n. caninum and toxoplasma gondii. based on the recognition of specif ...200515893073
prevalence of antibodies to toxoplasmagondii and neosporacaninum in red foxes (vulpesvulpes) from around the uk.the purpose of this study was to establish the prevalence of antibodies to toxoplasma gondii and neospora caninum in a random sample of red foxes from around the uk. lung fluid from over 500 foxes was examined using an indirect fluorescent antibody test. reciprocal titres of specific antibodies to t. gondii or n. caninum ranged from < 1:16 to 1:1024. a titre of 1:128 or greater was deemed indicative of exposure to the parasite. one hundred and eleven (20%) of the 549 foxes tested were seropositi ...200515893084
serological status for n. caninum, bovine viral diarrhea virus, and infectious bovine rhinotracheitis virus at pregnancy testing and reproductive performance in beef herds.neospora caninum, infectious bovine rhinotracheitis (ibr), and bovine viral diarrhea virus (bvdv) are important differentials for the diagnosis of infectious reproductive loss in beef herds. the objective of this study was to describe the serological status of both pregnant and non-pregnant beef cows from herds with varying levels of reproductive success. the study provided an opportunity to examine whether there were any associations between serological status for bvdv, ibr, and n. caninum and ...200515893892
seroepidemiology of toxoplasma gondii and neospora caninum in dogs from the state of paraíba, northeast region of brazil.a cross-sectional study was conducted to determine the seroprevalence of anti-toxoplasma gondii and anti-neospora caninum antibodies and to investigate the risk factors related to antibodies against t. gondii and n. caninum in dogs of the city of campina grande, state of paraiba, northeast region of brazil. for this purpose, 286 blood samples were collected from dogs during the rabies vaccination campaign, in september 2003, and on this occasion questionnaires addressing epidemiological aspects ...200415894024
toxoplasma gondii-like organisms in skin aspirates from a cat with disseminated protozoal 8-year-old, male domestic shorthair cat was referred to the matthew j. ryan veterinary hospital of the university of pennsylvania with a 3-day history of lethargy, inappetance, hyperemic skin nodules, coughing, and vomiting. laboratory results included nonregenerative anemia, lymphopenia, thrombocytopenia, hypoalbuminemia, hyponatremia, and increased alanine aminotransferase and aspartate aminotransferase activities. cytology of the skin nodules revealed many spindle- to crescent-shaped proto ...200515902669
neospora caninum: adoptive transfer of immune lymphocytes precipitates disease in balb/c mice.neospora caninum is a recently described apicomplexan parasite first isolated from a dog in 1988 and has subsequently been shown to infect a wide range of mammals. in mice, neospora can cause primary pneumonia, myositis, encephalitis, radiculoneuritis, and pancreatitis. whereas, certain aspects of the host immune response to toxoplasma gondii have been well studied, not as much is known about the full immune response to neospora. this paper examines whether or not immune splenocytes are able to ...200515916811
antibodies to neospora caninum in stray cats from north italy.antibodies to neospora caninum were determined in serum samples from 282 stray cats coming from four colonies near turin (north-west italy). sera were tested using a neospora agglutination test (nat). seroprevalence was 24.8% at 1:80, 12.8% at 1:160 and 5.3% at 1:320 dilution. seroprevalence in females and males, in different colonies and in different age classes, did not differ significantly. our results confirm that domestic cats are exposed to n. caninum and the observed seroprevalences sugge ...200515919155
the effects of ponazuril on development of apicomplexans in vitro.we examined the effects of 5 microg/ml ponazuril treatment on developing tachyzoites of neospora caninum and merozoites of sarcocystis neurona to better determine the mode of action of this anticoccidial drug. both parasites develop asexually by endogenesis. neospora caninum was selected for study because it develops by endodyogeny, which results in two tachyzoites being produced internally, and s. neurona was selected because it develops by endopolygeny which results in many merozoites being pr ...200515926999
exploration of immunoblot profiles of neospora caninum probed with different bovine immunoglobulin classes.the present study was attempted to compare the neospora caninum (n. caninum) antigenic bands recognized by different bovine immunoglobulin classes. a total 10, 5, 2, and 6 antigenic bands were exhibited on immunoblot profiles against bovine igm, ige, iga, and igg, respectively. a 46 kda band was probed as a common antigenic band except iga; 69 kda band was bovine igm and ige; 33, 37, 55, and 79 kda bands were bovine igm and igg; 72 kda band was found igm and iga profiles. based on the analysis, ...200515933437
effect of neosporosis on productive and reproductive performance of dairy cattle in costa rica.a cross-sectional study was conducted to assess the effect of neosporosis on productive and reproductive parameters in dairy cows. cows (n=2743) from 94 farms located in the most important dairy areas in costa rica were used in the study. the size of the herds ranged from 32 to 379 females (mean=110, median=125). an indirect elisa was used to determine the serostatus of the cows towards neospora caninum. the effect of neosporosis on milk production was analysed by a mixed linear model. in additi ...200515936812
expression of major histocompatibility complex class i and ii in a case of neospora caninum myositis in a dog.neosporosis was diagnosed in a 2-month-old dog by indirect immunofluorescence antibody test (ifat) and confirmed by means of histopathology and immunohistochemistry. the associated myositis was characterized by major histocompatibility complex expression on some muscle fibres. this finding indicates an immunological activation, of the muscle cells that, acquiring major histocompatibility complex expression, may, in some way, contribute to antigen presentation. a possible role of these glycoprote ...200415938572
seronegative conversion in four neospora caninum-infected cows, with a low rate of transplacental transmission.four neospora-seropositive pregnant cows (prebreeding indirect fluorescent antibody (ifa) titers between 1:400 and 1:1600) were confined and observed until parturition. all cows gave birth to normal calves. selected tissues were tested for nc by histopathology, immunohistochemical (ihc) and polymerase chain reaction (pcr). parasite isolation was attempted in vero cell cultures. at parturition, all cows were seronegative at 1:200 and two of four cows had a titer of 1:100 when further tested. thre ...200515939537
evaluation of three enzyme-linked immunosorbent assays for detection of antibodies to neospora caninum in bulk milk.three elisas for the detection of antibodies against neospora caninum in bulk milk were evaluated in 162 dutch dairy herds. the first elisa was the dutch animal health service (ahs) in-house elisa, developed from the routine in-house serum elisa. the other two elisas were commercial milk elisas from idexx and lsi. blood samples of all lactating cows in 162 dairy herds were tested using the ahs in-house serum elisa. based on previous studies in the netherlands a within-herd n. caninum seroprevale ...200515970385
hplc purification of recombinant ncgra6 antigen improves enzyme-linked immunosorbent assay for serodiagnosis of bovine neosporosis.the gene for a dense granule protein (ncgra6) of neospora caninum was expressed in escherichia coli as a his-tag fusion protein and purified by ninta affinity chromatography. in a preliminary study, high binding of antibodies from n. caninum-negative cows was observed in enzyme-linked immunosorbent assay (elisa) using ninta-purified ncgra6. analysis of ninta eluates revealed a significant number of e. coli proteins that co-purified with recombinant ncgra6. in an attempt to improve the relative s ...200515970387
effects of seropositivity for bovine leukemia virus, bovine viral diarrhoea virus, mycobacterium avium subspecies paratuberculosis, and neospora caninum on culling in dairy cattle in four canadian provinces.the purpose of this research was to determine the effects of seropositivity for exposure to bovine leukemia virus (blv), bovine viral diarrhoea virus (bvdv), mycobacterium avium subspecies paratuberculosis (map) and neospora caninum (nc) on overall and reason-specific culling in canadian dairy cattle. serum samples from, approximately, 30 randomly selected cows from 134 herds were tested for antibodies against blv, map and nc using commercially available elisa test kits, while 5 unvaccinated cat ...200515970402
culture of neospora caninum in the presence of a mycoplasma removal agent results in the selection of a mutant population of tachyzoites.mycoplasmas are common contaminants of eukaryotic cells grown in tissue culture. a commercially available mycoplasma removal agent (mra) was therefore assessed for its effect on tachyzoites of neospora caninum, in order to determine its suitability for further use in parasite cell cultures. analyses of tachyzoite and excreted-secreted proteins and antigens by sds-page and western blotting show that mra treatment results in the rapid selection of a mutant population that differs from the control ...200515977896
confirmed clinical neospora caninum infection in a boxer puppy from argentina.generalized neosporosis was diagnosed in a 2-month-old boxer puppy. the dog had a history of progressive paralysis and muscle atrophy, followed by cervical weakness, stiff jaws and dysphagia. the dog had a 1:12,800 antibody titer for neospora caninum and was negative for antibodies to toxoplasma gondii by the indirect fluorescent antibody test (ifat). after euthanasia a complete necropsy was carried out. the puppy had a megaesophagus. microscopically, tachyzoites and tissue cysts were observed i ...200515978728
recombinant nhsag1 elisa: a sensitive and specific assay for detecting antibodies against neospora hughesi in equine serum.neospora hughesi is a recently identified cause of equine protozoal myeloencephalitis. however, the significance of this parasite is poorly understood. an enzyme-linked immunosorbent assay (elisa) with a recombinant form of the n. hughesi 29-kda surface antigen (rnhsag1) was developed for serodiagnosis of equine n. hughesi infections. parallel elisa analysis showed that animals immunized or infected with n. hughesi exhibited greater antibody reactivity with rnhsag1 than with the neospora caninum ...200515986623
identification and characterization of cross-reactive antigens from neospora caninum and toxoplasma gondii.murine monoclonal antibodies (mabs) against neospora caninum tachyzoites were produced to identify the cross-reactive antigens between n. caninum and toxoplasma gondii. ten mabs recognizing cross-reactive antigens of both parasites were obtained and tentatively classified into 6 different groups based on their reactivity patterns in an indirect fluorescent antibody test and western blot analysis. three mabs in group 1 recognized antigens located on the surface of parasites with molecular masses ...200515991490
development of rapid immunochromatographic test with recombinant ncsag1 for detection of antibodies to neospora caninum in immunochromatographic test (ict) with recombinant surface antigen 1 of neospora caninum (ncsag1) was developed for the rapid detection of antibodies to n. caninum in cattle. the ict was used to clearly discriminate between immunofluorescent-antibody test (ifat)-positive bovine sera and ifat-negative bovine sera. serum samples collected from cattle in yanbian, china, were examined by the ict. of the 96 serum samples, 23 (24.0%) were positive by the ict, and 19 (19.8%) samples were positive by ...200516002641
influence of the stage of pregnancy on neospora caninum distribution, parasite loads and lesions in aborted bovine the present work we have studied in neospra caninum aborted bovine foetuses the influence of foetal age (first, second and third gestational periods) on parasite distribution by nested pcr, parasite loads by real-time pcr and n. caninum associated lesions. for this purpose, a total of 220 aborted foetuses were analysed and detection of n. caninum infection was accomplished by nested-pcr in brain, heart and liver, detecting the presence of the parasite in 72 (32.7%) bovine foetuses. when the d ...200616023188
phylogenetic congruence of sarcocystis neurona dubey et al., 1991 (apicomplexa: sarcocystidae) in the united states based on sequence analysis and restriction fragment length polymorphism (rflp).the objectives of the present study were to assess the genetic diversity, phylogeny and phylogeographical relationships of available sarcocystis neurona isolates from different localities in the united states. all 13 sarcocystis isolates from different hosts were subjected to polymerase chain reaction restriction fragment length polymorphism (pcr-rflp) analyses using two published dna markers (25/396 and 33/54). the 334 bp sequence of the 25/396 marker of these isolates and besnoitia darlingi, b ...200516025209
effects of host maturity and prior exposure history on the production of neospora caninum oocysts by investigate whether dogs shed neospora caninum oocysts more than once, five dogs with a previous history of shedding oocysts were fed infected bovine tissues. two of three dogs shed oocysts when they were re-exposed 18-20 months after the first challenge; two other dogs re-exposed earlier, only 8 months after the primary exposure, did not produce oocysts. these results suggest that dogs may become refractory to shedding n. caninum oocysts for a period approximately between 8 and 18 months aft ...200516029931
neospora abortion in new zealand cattle.protozoal abortion in cattle was retrospectively studied by examining material submitted to the batchelar animal health laboratory in the years 1987-90. only cases in which foetal brain had been submitted were examined. histologically, protozoal lesions were seen in the brains of 28% of 320 aborted foetuses. lesions were also seen in 10% of the hearts and 34% of the placentas examined, but these were not as characteristic as those in the brains. protozoa, confirmed immunohistochemically as being ...199116031639
protozoal encephalomyelitis of dogs involving neospora caninum and toxoplasma gondii in new a retrospective study of 15 cases of encephalomyelitis in dogs, three cases of neospora caninum and two cases of toxoplasma gondii infection were identified using immunohistochemical staining of central nervous system sections. all cases of neosporosis showed ataxia and progressive hind limb paralysis due to multifocal non-suppurative meningoencephalomyelitis which was most severe in the spinal cord and base of the brain stem. neospora tissue cysts could not be distinguished morphologically f ...199716031995
serology of a neospora abortion outbreak on a dairy farm in new zealand: a case describe the kinetics of serological titres after an abortion outbreak in april-may 1995 due to neospora caninum affected 17 dairy cows in a herd of 320.199816032006
neospora caninum infection in an aborted bovine foetus in brazil. 199916032066
maintenance of neospora caninum tachyzoites using mongolian gerbils (meriones unguiculatus). 199916032067
prevalence of neospora antibodies in beef cattle in new estimate the prevalence of neospora infection in a sample of new zealand beef cattle.200016032142
recent advances in understanding the epidemiology of neospora caninum in cattle.introduction: neospora caninum is the most frequently diagnosed cause of cattle abortion in new zealand and is an important pathogen worldwide. the parasite has been found in aborted bovine foetuses and in puppies with neurological disease. recent developments have provided new insights into the epidemiology of the disease that are reviewed in this article. transmission: transplacental infection is of major importance in the spread of n. caninum overseas. most congenital infections result in the ...200116032161
control options for neospora caninum infections in cattle--current state of knowledge.this article reviews control options for neospora caninum infection and abortion in cattle, drawing on published literature and the authors' own research in this field. apart from the successful use of embryo transfer to prevent congenital infection in calves born to infected cows, there are currently no accepted control methods for the prevention of abortions in cattle. the epidemiological data at hand suggest that concomitant infections with bovine pestivirus increase the risk of abortion sign ...200216032217
hammondia heydorni oocysts in the faeces of a greyhound in new identify oocysts found in faecal material of a greyhound.200316032288
prevalence of antibodies to neospora caninum in dogs of rural or urban origin in central new investigate the seroprevalence of neospora caninum infection in populations of dogs from dairy farms, sheep/beef farms and urban areas in the central part of new zealand. it was postulated seroprevalence would be higher for farm dogs than urban dogs if the life-cycle of this parasite involves transmission between dogs and cattle.200316032332
identification and characterization of neospora caninum cyclophilin that elicits gamma interferon production.gamma interferon (ifn-gamma) response is essential to the development of a host protective immunity in response to infections by intracellular parasites. neosporosis, an infection caused by the intracellular protozoan parasite neospora caninum, is fatal when there is a complete lack of ifn-gamma in the infected host. however, the mechanism by which ifn-gamma is elicited by the invading parasite is unclear. this study has identified a microbial protein in the n. caninum tachyzoite n. caninum cycl ...200516041025
immunization with native surface protein ncsrs2 induces a th2 immune response and reduces congenital neospora caninum transmission in mice.ncsrs2, a tachyzoite surface protein of neospora caninum, is an immunodominant protein with respect to induction of antibody production and has a role in attachment and invasion of host cells. native ncsrs2 was isolated from whole tachyzoite lysate antigen by affinity chromatography using ncsrs2 specific monoclonal antibody and used to immunize balb/c mice in a congenital transmission study. ncsrs2 was a highly conserved protein as indicated by comparison of deduced amino acid sequence obtained ...200516051244
occurrence of anti-neospora caninum antibodies in water buffaloes (bubalus bubalis) from the northern region of brazil.anti-neospora caninum antibodies were determined in sera of 196 water buffaloes (bubalus bubalis) from three farms of the northern region of brazil, using an indirect fluorescent antibody test. antibody titers were found in 139 (70.9%) buffaloes with dilution values ranging from > or =25 to <200 in 41 animals, > or =200 to <800 in 35 animals, and > or =800 in 63 animals. the number of animals presenting titers > or =800 was statistically higher (p<0.05). all farms presented positive animals, how ...200516051440
the host-parasite relationship in bovine neosporosis.infection with the protozoan parasite neospora caninum is thought to be a major cause of reproductive failure in cattle worldwide. cattle infected with the parasite are three to seven times more likely to abort compared to uninfected cattle. the parasite may be transmitted to cattle through the ingestion of oocysts that are shed in the faeces of acutely infected dogs (definitive host of n. caninum) or by congenital infection from mother to foetus via the placenta. interestingly, transplacental t ...200516098610
comparison of proteome and antigenic proteome between two neospora caninum isolates.this study was conducted to explore the relationship between two isolates of neospora caninum (n. caninum) (kba-2 and vmdl-1) using proteomics. to achieve the goal, proteins of n. caninum tachyzoite lysates of kba-2 and vmdl-1 were separated by two-dimensional gel electrophoresis (2-de), stained with silver-nitrate and analyzed using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (maldi-tof ms) to compare protein profiles. in addition, proteins separated by 2-de wer ...200516098674
neospora caninum seropositivity and association with abortions in dairy cows in mexico.the seroprevalence of neospora caninum infection was estimated from a sample of 813 cattle from 20 dairy herds in five regions in mexico. the true prevalence of infection was estimated to be 42% (95% ci: 39, 46). seropositivity was associated with abortion (odds ratio (or)=2.0) and was higher among cows raised on-farm (41%), than among replacement cattle purchased outside the farm (28%). the ors relating abortion to seropositivity in individual herds ranged from 1.3 to 10. overall, 26% of the ab ...200516098675
the sylvatic cycle of neospora caninum: where do we go from here?bovine abortions due to neospora caninum infection are a major cattle-production problem worldwide. the parasite is readily maintained in cattle populations by vertical transmission. the domestic dog excretes oocysts in its feces and, after sporulation, these oocysts are infectious to cattle. current control measures are aimed at culling infected cows and limiting the access of cattle to infective oocysts. the recent revelations that coyotes (canis latrans) can excrete n. caninum oocysts in thei ...200516098812
prevalence of toxoplasma gondii and neospora caninum in sika deer from eastern hokkaido, japan.brain and serum were collected from 120 and 12 free-ranging sika deer (cervus nippon yesoensis), respectively, from six regions in eastern hokkaido during controlled hunts in the autumn of 2003. brains were tested for neospora caninum and toxoplasma gondii dna by polymerase chain reaction (pcr) assays. antibodies to toxoplasma gondii were measured by means of a latex agglutination test. no brain tested positive for either type of dna, and no antibody to toxoplasma gondii was detected in serum, s ...200516107684
adaptation of neospora caninum isolates to cell-culture changes: an argument in favor of its clonal population structure.neospora caninum, a recently recognized protozoan parasite of animals, is considered to be a major cause of bovine abortion worldwide. although its life cycle is not completely known, recent studies suggest that the sexual stage occurs in dogs. the prevalence of sexual reproduction in n. caninum, however, is unknown. we investigated the ability of 3 n. caninum isolates (nc-1, nc-sweb1, and nc-liverpool) to propagate asexually for approximately 250 parasite generations in a cell line in which the ...200516108539
neospora caninum antigens defined by antigen-dependent bovine cd4+ t cells.neosporosis is an important cause of pregnancy loss in cattle worldwide. the objective of the present study was to identify neospora caninum antigens as vaccine candidates using antigen-specific, short-term cd4+ t cells established from n. caninum-immunized and -challenged cows. whole n. caninum tachyzoite lysate was separated into 6 fractions by deae anion-exchange chromatography using high-pressure liquid chromatography (hplc). the cd4+ t-cell proliferation assay results indicated that antigen ...200516108548
reduced infection and protection from clinical signs of cerebral neosporosis in c57bl/6 mice vaccinated with recombinant microneme antigen ncmic1.ncmic1 is a 460 amino acid neospora caninum microneme protein implicated in host cell adhesion and invasion processes. in this study, we assessed the potential protectivity of ncmic1-based vaccination against experimental n. caninum infection in mice, employing both recombinant antigen vaccines and dna vaccines. recombinant ncmic1 (recncmic1) was expressed in escherichia coli as gluthatione-s-transferase-fusion protein. the corresponding ncmic1 cdna was cloned into the pcdna3.1 expression plasmi ...200516108562
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 ...200516108816
environmental effect on the occurrence of anti-neospora caninum antibodies in pampas-deer (ozotoceros bezoarticus).sera obtained from pampas-deer (ozotoceros bezoarticus) captured in two different brazilian environments were analyzed for the presence of anti-neospora caninum antibodies by the indirect fluorescent antibody test (ifat). samples were collected from 23 animals from a savanna area in the national park of emas, in the state of goiás, brazil. this area is surrounded by cultivated lands and allows very little contact between wild and domestic animals. another batch of samples was collected from 16 a ...200516112811
in vitro efficacies of nitazoxanide and other thiazolides against neospora caninum tachyzoites reveal antiparasitic activity independent of the nitro group.the thiazolide nitazoxanide [2-acetolyloxy-n-(5-nitro-2-thiazolyl)benzamide] (ntz) exhibits a broad spectrum of activities against a wide variety of intestinal and tissue-dwelling helminths, protozoa, and enteric bacteria infecting animals and humans. the drug has been postulated to act via reduction of its nitro group by nitroreductases, including pyruvate ferredoxin oxidoreductase. in this study, we investigated the efficacies of nitazoxanide and a number of other thiazolides against neospora ...200516127045
neospora caninum protein disulfide isomerase is involved in tachyzoite-host cell interaction.we have previously shown that treatment of neospora caninum tachyzoites with the aspartyl protease inhibitor pepstatin a reduces host cell invasion [naguleswaran, a., muller, n., hemphill, a., 2003. neospora caninum and toxoplasma gondii: a novel adhesion/invasion assay reveals distinct differences in tachyzoite-host cell interactions. exp. parasitol. 104, 149-158]. pepstatin a-affinity-chromatography led to the isolation of a major band of approximately 52 kda which was identified as a homologu ...200516129440
toxoplasma gondii dense granule protein 3 (gra3) is a type i transmembrane protein that possesses a cytoplasmic dilysine (kkxx) endoplasmic reticulum (er) retrieval motif.studies using antibodies to immunolocalize the toxoplasma gondii dense granule protein gra3, have shown that this protein associates strongly with the parasitophorous vacuole membrane (pvm). however, as there was no predicted membrane-spanning domain this highlighted an unanswered paradox. we demonstrate that the previously published sequence for gra3 is actually an artificial chimera of 2 proteins. one protein, of molecular weight 65 kda, shares the c-terminus with published gra3 and possesses ...200516149193
differential detection of hammondia hammondi from toxoplasma gondii using polymerase chain reaction.hammondia hammondi and toxoplasma gondii are two related coccidian parasites, with cats as definitive hosts and warm-blooded animals as intermediate hosts. it is difficult to differentiate them by morphological and serological parameters. in the present study, primers were designed to specifically amplify the its-1 region of h. hammondi to differentiate it from t. gondii. attempts were made to detect the presence of h. hammondi dna in the tissues of mice infected with h. hammondi alone, as well ...200516153883
subcellular fractionation and molecular characterization of the pellicle and plasmalemma of neospora caninum.a characteristic structural feature of toxoplasma gondii and neospora caninum is the presence of a triple-membrane pellicle, on the zoite stages of their complex life-cycle. here we report the results of electron microscopic studies which show that the pellicle is made of a typical plasmalemma covered on its cytoplasmic side by a system of flattened vesicles named the inner membrane complex. using methods described previously for the purification of pellicle and plasmalemma fractions from t. gon ...200516174411
oocysts of neospora caninum, hammondia heydorni, toxoplasma gondii and hammondia hammondi in faeces collected from dogs in germany.faecal samples of 24,089 dogs were examined coproscopically in two veterinary laboratories in germany between march 2001 and october 2004. in 47 dogs, oocysts of 9-14 microm size were found. their morphology was similar to those of hammondia heydorni and neospora caninum. samples of 28 of these dogs were further examined by inoculation into gerbils: seven isolates induced a specific antibody response against antigens of n. caninum nc-1 tachyzoites. this response suggests that the isolates contai ...200516197949
nationwide seroprevalence of neospora caninum among dairy cattle in japan.serum samples from 2420 clinically healthy dairy cattle, randomly selected from stored sera in 18 districts of japan, were tested for the presence of neospora caninum antibodies using an indirect fluorescent antibody test (titer > or =1:200). nationwide seroprevalence is estimated at 5.7% (139/2420). seropositive cattle were detected in all surveyed districts despite the evidence of confirmed case reports of bovine neosporosis, showing that n. caninum is widely distributed throughout japan. age- ...200616207513
endogenous and exogenous transplacental infection in neospora caninum and toxoplasma is clear from researching the vertical transmission of neospora caninum in cattle that the terms 'vertical', 'congenital' and, indeed, 'transplacental' are inadequate for describing two extremely different situations that have fundamentally different immunological, epidemiological and control implications. a similar situation pertains to toxoplasma gondii in different hosts. we advocate the use of the terms 'endogenous transplacental infection (tpi)' to define foetal infection from a recrudes ...200516223599
association between neospora caninum antibodies and blue tongue vaccination in dairy cows. 200516244963
inhibitory activities of epidermal growth factor receptor tyrosine kinase-targeted dihydroxyisoflavone and trihydroxydeoxybenzoin derivatives on sarcocystis neurona, neospora caninum, and cryptosporidium parvum development.several gene sequences of parasitic protozoa belonging to protein kinase gene families and epidermal growth factor (egf)-like peptides, which act via binding to receptor tyrosine kinases of the egf receptor (egfr) family, appear to mediate host-protozoan interactions. as a clue to egfr protein tyrosine kinase (ptk) mediation and a novel approach for identifying anticoccidial agents, activities against sarcocystis neurona, neospora caninum, and cryptosporidium parvum grown in bm and hct-8 cell cu ...200516251305
attachment and invasion of toxoplasma gondii and neospora caninum to epithelial and fibroblast cell lines in vitro.attachment and invasion of toxoplasma gondii and neospora caninum to a cat and a dog fibroblast cell line and 2 epithelial cell lines (a cat kidney and vero) were compared in vitro using fluorescence antibody methodology. in addition, trypsin treatment of tachyzoites was used to determine whether protein molecules were essential to the process of invasion. the results show that both t. gondii and n. caninum invaded all 4 cell lines, and that pre-treatment of t. gondii tachyzoites with trypsin ca ...200516255816
intrauterine neospora caninum inoculation of, we studied the potential of neospora caninum tachyzoites to infect heifers when administered in utero by artificial insemination via contaminated semen. eighteen primiparous cyclic heifers were hormonally synchronized and artificially inseminated. nine of them, which were inseminated with semen containing 10(7) live n. caninum nc-1 isolate-tachyzoites, reacted with seroconversion and a specific ifn-gamma response. moreover, n. caninum dna was demonstrated by a nested-pcr in the blood of al ...200616288958
comparison of different methods for the solubilisation of neospora caninum (phylum apicomplexa) antigen.parasite antigens are of interest both for the development of better diagnostic tools and potential subunit vaccines. particularly, relevant in this regard are membrane proteins as the first point of contact between host and parasite. here, four different methods for the extraction and solubilisation of neospora caninum proteins were evaluated by comparing protein yield, specific antigenicity and relative protein abundance in the electrophoresis profile. extraction with sds and sulphobetaines (s ...200616289567
seroepidemiology of neospora caninum infection in dairy cattle herds in mashhad area, iran.a seroepidemiology study of neospora caninum was conducted in mashhad area, khorasan province, iran on 337 cattle from 30 dairy using elisa. all herds had seropositive cattle and antibodies were formed in 46% of the cattle. abortion was significantly associated with seropositivity of cattle. the highest risk of abortion was observed in the 1-2-year-old cows. our results indicate that neospora infection is widespread in iran.200616289861
validation of a commercially available celisa test for canine neosporosis against an indirect fluorescent antibody test (ifat).a commercially available competitive enzyme-linked immunosorbent assay (celisa, vmrd) was validated for the detection of neospora caninum antibodies in the serum of dogs, using as a reference test an indirect fluorescent antibody test (ifat, fuller). a partial verification approach was used. a total of 618 dogs were screened with celisa and a subset of positive and negative sera (n=237) were then tested with ifat. naïve relative sensitivity (se(nv)) and naïve relative specificity (sp(nv)) of cel ...200616293328
identification and molecular cloning of the neospora caninum sag4 gene specifically expressed at bradyzoite, we identify and clone the ncsag4 gene, orthologue to the toxoplasma gondii tgsag4 gene, and the first reported gene to be expressed specifically during the neospora caninum bradyzoite stage. to isolate ncsag4, we designed degenerate oligonucleotides based on the tgsag4 protein amino acid sequence. a 312-bp dna fragment was amplified by pcr from n. caninum genomic dna, whose sequence showed 65% identity to tgsag4 gene over 257 bp. ncsag4 gene sequence was obtained by pcr genome walking. nuc ...200616303187
occurrence of neospora caninum in dogs and its correlation with visceral leishmaniasis in the urban area of campo grande, mato grosso do sul, brazil.neospora caninum is an obligate intracellular protozoan that can infect domestic and wild canids, as well as ruminants and equines, and is described as causing neuromuscular alteration and death in dogs. visceral leishmaniasis (vl) is an infectious disease that affects both humans and animals, being caused by protozoan parasites of the genus leishmania, of which leishmania (leishmania) chagasi is found in brazil -- transmitted by sand flies, such as lutzomyia longipalpis, in most of the american ...200616310954
prevalence of sarcocystis neurona and neospora spp. infection in horses from brazil based on presence of serum antibodies to parasite surface antigen.sera from 961 horses from brazil were tested for antibodies against the major surface antigens snsag4 and nhsag1 to determine the seroprevalence of sarcocystis neurona and neospora hughesi, respectively. antibodies against snsag4 were detected in 669 (69.6%) of the horses, while antibodies against nhsag1 were detected in only 24 (2.5%) of the horses. these serologic results suggest that there is a high concentration of s. neurona in the environment of brazil, which results in marked exposure of ...200616310955
use of avidity enzyme-linked immunosorbent assay and avidity western blot to discriminate between acute and chronic neospora caninum infection in cattle.avidity serological tests (avidity enzyme-linked immunosorbent assay [elisa] and avidity western blot) were developed and used to differentiate between acute (primary infection, reinfection, and recrudescence) and chronic neospora caninum infection in cattle. in addition, the pattern of immunoglobulin g (igg) avidity maturation against different specific antigens of n. caninum tachyzoites was studied. sequential serum samples were collected from cattle naturally and experimentally infected with ...200516312235
persistence of neospora caninum-specific immunoglobulin g antibodies in bovine blood and lung tissue stored at room temperature.blood and tissue samples obtained at necropsy from carcasses showing varying degrees of postmortem decomposition are sometimes used as sources of antibodies for serological tests, primarily in studies involving wild animals or in examination of aborted fetuses. to test the persistence of antibodies in blood and lung tissue kept at room temperature (approximately 20degreesc) for up to 48 and 118 days, respectively, serum and lung fluid from neospora caninum-infected and uninfected cattle were tes ...200516312237
prevalence of neospora caninum infection in sardinian dairy farms (italy) detected by iscom elisa on tank bulk milk.neospora caninum is a heteroxenous cyst-forming coccidian closely related to toxoplasma gondii and is considered one of the major causes of abortions in cattle worldwide. the present work aims to update the epidemiological trend of n. caninum of dairy cattle in sardinia island, western mediterranean (italy). for this reason, we used the newest enzyme-linked immunoassay (elisa) methodology that exploits immune-stimulating complexes (iscoms) principle and allows us to point out the infection in th ...200616323023
proportion of abortions due to neosporosis among dairy cattle in japan.a retrospective cohort study was conducted to clarify the association between seropositive reactions to neospora caninum and subsequent reproductive disorders among dairy cattle in japan. a statistically significant association between neospora seropositive reactions and abortions was observed (p=0.016), and seropositive cattle were 6.1 times more likely to abort compared to seronegative cows. no significant differences were observed between seropositive reactions and other reproductive disorder ...200516327231
neospora caninum: infection induced il-10 overexpression in rat astrocytes in vitro.the effect of neospora caninum, a parasite that causes abortion and neuromuscular changes, has been investigated on a major population of neural cells, the astrocytes. highly enriched astroglial primary cultures obtained from neonatal rats were infected after 21 days of culture. astroglial reactivity, il-10 and ifn-gamma expression, and cell viability (lactate dehydrogenase activity, metabolization of tetrazolium salt, and trypan blue exclusion assay) have been investigated after 24 and 72 h of ...200616332369
survey of neospora caninum infection by nested pcr in aborted fetuses from dairy farms in aguascalientes, mexico.the aim of this study was to determine by nested pcr probe the presence of neospora caninum in fetal brain tissue from aborted dairy fetuses in aguascalientes, a state in the central part of mexico. forty-four fetal brains from eight dairy farms were analyzed using single tube nested pcr probe with primers nf1, ns2, nr1 and sr1, with histopathology as the reference technique. from the 44 bovine fetuses studied, 35 (80%) were diagnosed as being infected by n. caninum using pcr probe and 20 (45%) ...200616332413
development of a practical immunochromatographic test with recombinant p50 for the diagnosis of babesia gibsoni infection in immunochromatographic test (ict), using recombinant truncated p50 (p50t), for the detection of antibodies to babesia gibsoni was developed and evaluated. whereas all sera from specific pathogen-free dogs were clearly negative, all sera from dogs experimentally infected with b. gibsoni were clearly positive in the ict. in addition, the ict detected no cross-reactivity with sera from dogs experimentally infected with closely related parasites, b. canis canis, b. canis vogeli, and b. canis rossi ...200516336730
herd-level risk factors for neospora caninum seroprevalence in dairy farms in southern brazil.a cross-sectional study was used to test the relationship between herd seroprevalence to neospora caninum and various potential herd-level risk factors in 60 dairy farms located in two distinct regions in southern brazil. thirty farms were randomly selected from within each region. a questionnaire was designed to summarize each farm's production system as it might relate to n. caninum transmission. the questionnaire contained 105 closed questions relating to general characteristics of the farms, ...200616343669
neospora caninum: antibodies directed against tachyzoite surface protein ncsrs2 inhibit parasite attachment and invasion of placental trophoblasts in vitro.polyclonal and monoclonal antibodies to native neospora caninum tachyzoite surface protein ncsrs2 were generated and tested in vitro for their ability to neutralize tachyzoite attachment to and invasion of host cells. host cells included vero cells and a newly cloned, immortalized ovine trophoblast cell line obtained from primary cultures of ovine placenta. the ovine trophoblasts had morphology consistent with fetal trophoblasts and expressed mrna for interferon-tau, a marker for trophoblasts. n ...200616375893
seroprevalences of antibodies to neospora caninum and toxoplasma gondii in zoo animals.neospora caninum is an apicomplexan parasite that causes neuromuscular disease in dogs and abortions in cattle. little is known about the prevalence of antibodies to this parasite in zoo animals. sera from 556 animals, from 13 czech and slovak zoos were tested for antibodies to n. caninum and toxoplasma gondii by indirect fluorescent antibody test. antibodies to n. caninum were found in 31 of 556 zoo animals (5.6%), representing 18 of 114 species tested: eurasian wolf (canis lupus lupus), maned ...200616387445
the prevalence of neospora caninum and co-infection with toxoplasma gondii by pcr analysis in naturally occurring mammal populations.neospora caninum and toxoplasma gondii are closely related intracellular protozoan parasites associated with bovine and ovine abortion respectively. little is known about the extent of neospora/toxoplasma co-infection in naturally infected populations of animals. using nested pcr techniques, based on primers from the nc5 region of n. caninum and sag1 for t. gondii, the prevalence of n. caninum and its co-infection with t. gondii were investigated in populations of mus domesticus, rattus norvegic ...200616393351
application of repeated bulk milk testing for identification of infection dynamics of neospora caninum in thai dairy this prospective study, repeated bulk milk tests were evaluated to establish the neospora caninum antibody status and to describe infection dynamics and changes in prevalence of n. caninum in thai dairy herds. bulk milk from 418 herds in northeast thailand were collected at three consecutive samplings, in december 2002 (sampling 1), april (sampling 2) and december 2003 (sampling 3). all samples were analysed for presence of n. caninum antibodies by iscom elisa, and the optical density values ...200616406330
evaluation of an iscom elisa used for detection of antibodies to neospora caninum in bulk milk.the intracellular parasite neospora caninum is increasingly recognized as an important cause of abortion and stillbirth in cattle. presence of specific antibodies indicates infection, and the immunostimulating complex (iscom) enzyme-linked immunoassay (elisa) has previously been evaluated for use on individual milk and sera. in the present study, this test is investigated for use on bulk milk. in this study, 124 herds were used to analyse the relationship between within-herd prevalences based on ...200616417934
supranational comparison of neospora caninum seroprevalences in cattle in germany, the netherlands, spain and sweden.herd, within-herd and animal prevalences for neospora caninum in beef and dairy cattle were compared between four countries. in randomly selected herds from regions of germany, the netherlands, spain and sweden that were representative for the cattle production of these countries, all animals > or = 2 years were examined serologically by enzyme-linked immunosorbent assays (elisas) with high test specificity (> 98.0%). in a previous study, the elisas had been validated against each other. single ...200616417972
evaluation of homologous, heterologous, and affinity conjugates for the serodiagnosis of toxoplasma gondii and neospora caninum in maned wolves (chrysocyon brachyurus).use of serological tests in the diagnosis of infectious diseases in wild animals has several limitations, primarily the difficulty of obtaining species-specific reagents. wild canids, such as maned wolves (chrysocyon brachyurus), are highly predisposed to infection by toxoplasma gondii and, to a lesser extent, to neospora caninum. the aim of the present study was to evaluate homologous, heterologous, and affinity conjugates in enzyme-linked immunosorbent assays (elisas) and indirect fluorescent ...200516419770
prevalence of antibodies to neospora caninum in wild animals.antibodies to neospora caninum were determined in several species of wild animals in the united states by the neospora agglutination test (nat). antibodies (nat 1:40 or higher) were found in 5 of 249 bison (bison bison), 5 of 160 caribou (rangifer tarandus), 4 of 162 moose (alces alces), 4 of 122 wolves (canis lupus), and 1 of 224 musk ox (ovibos moschatus) but not in 197 black bears (ursus americanus). to our knowledge, this is the first report of antibodies to n. caninum in bison and caribou. ...200516419771
serological evidence of neospora caninum in dual-purpose cattle herds in venezuela.bovine neosporosis is a parasitic disease produced by neospora caninum that induces abortion in cows, and consequently has a negative impact on the herd's reproductive efficiency. the main objective of this research was to determine the serological evidence of n. caninum in cattle herds from venezuela using an indirect antibody capture elisa test. four hundred and fifty-nine (459) serum samples from crossbred adult cows were collected to be tested for neospora antibodies. the sampled cows came f ...200616423467
seroprevalence of neospora caninum and toxoplasma gondii in camels (camelus dromedarius) in mashhad, hundred twenty camels were blood-sampled and used to evaluate serological screening for neospora caninum and toxoplasma gondii infection by indirect fluorescent antibody test (ifat) in mashhad, iran, during years 2004-2005. of the 120 camels, antibodies to n. caninum were found in three in titers of 1:20 and in four in titers of 1:40 using whole n. caninum tachyzoites as ifat slide (vmrd inc., pullman, wa 99163, usa). antibodies to t. gondii were found in three camels in titers 1:20 and in t ...200616425066
detection of immunoglobulin g antibodies to neospora caninum in humans: high seropositivity rates in patients who are infected by human immunodeficiency virus or have neurological disorders.considering that little is known about the epidemiology of neospora caninum infection in humans, particularly in populations with high toxoplasma gondii infection rates, the present study aimed to investigate the presence of antibodies to n. caninum in t. gondii-seropositive and -seronegative individuals. a total of 256 serum samples divided into four groups (61 samples from human immunodeficiency virus [hiv]-positive patients, 50 samples from patients with neurological disorders, 91 samples fro ...200616426004
the protozoan neospora caninum directly triggers bovine nk cells to produce gamma interferon and to kill infected fibroblasts.natural killer (nk) cells are considered to be key players in the early innate responses to protozoan infections, primarily indirectly by producing gamma interferon (ifn-gamma) in response to cytokines, like interleukin 12 (il-12). we demonstrate that live, as well as heat-inactivated, tachyzoites of neospora caninum, a toxoplasma-like protozoan, directly trigger production of ifn-gamma from purified, il-2-activated bovine nk cells. this response occurred independently of il-12 but was increased ...200616428740
low seroprevalence of antibodies to neospora caninum in wild canids in israel.the role of domestic dogs in the epidemiology of neospora caninum as well as the relationship between n. caninum infection of farm dogs and cattle were demonstrated, however, evidence is scarce regarding the role of wild canids in domestic animal neosporosis. the present study was undertaken to evaluate the role of wild canids in the epidemiology of bovine neosporosis in israel by analyzing the prevalence of antibodies to n. caninum in wild canids. sera samples were collected from 114 free rangi ...200616436314
protecting babies: vaccine strategies to prevent foetopathy in neospora caninum-infected cattle.neospora caninum is an apicomplexan protozoan parasite that is a significant infectious abortifacient agent in cattle. despite the fact that it is a member of a well described taxonomic group, it is a relatively newly discovered parasite and its biology is not yet fully understood. cattle become infected either congenitally via transplacental transmission or post-natally by ingesting oocysts derived from the definitive host; dogs and coyotes are the only definitive hosts that have been described ...200616441503
clinical sarcocystis neurona, sarcocystis canis, toxoplasma gondii, and neospora caninum infections in dogs.sarcocystis neurona, sarcocystis canis, toxoplasma gondii, and neospora caninum are related apicomplexans that can cause systemic illness in many species of animals, including dogs. we investigated one breeder's 25 basset hounds for these infections. in addition, tissues from dogs and other non-canine hosts previously reported as s. canis infections were studied retrospectively. schizonts resembling those of s. neurona, and recognized by polyclonal rabbit anti-s. neurona antibodies, were found i ...200616458431
neospora caninum: serological follow-up in dairy cows during pregnancy.we conducted a longitudinal study to follow-up the anti-neospora caninum serologic status in 30 initially seropositive and 83 initially seronegative cows during their pregnancy. study cows were blood-sampled every other month during pregnancy until parturition. blood serum samples were screened for anti-n. caninum antibodies by elisa. cows that seroconverted were re-tested by immunoblot as a confirmation test. among 30 seropositive cows, 28 cows remained seropositive during the whole pregnancy, ...200616469449
pcr detection of neospora caninum, toxoplasma gondii and encephalitozoon cuniculi in brains of wild carnivores.neospora caninum, toxoplasma gondii and encephalitozoon cuniculi are important pathogens with affinity to the central nervous system of many animals. 240 brains of wild carnivores were examined by pcr-based diagnosis. the presence of n. caninum dna was confirmed in 4.61% (7/152) red foxes (vulpes vulpes). dna of t. gondii was found in 4.92% (3/61) martens (martes sp.) and in 1.32% (2/152) red foxes. dna of e. cuniculi was determined in 3.28% (2/61) martens and in one examined european otter (lut ...200616472923
use of a multiplex polymerase chain reaction to rapidly differentiate neospora caninum from toxoplasma gondii in an adult dog with necrotizing myocarditis and myocardial infarct.this report describes a 3-year-old male castrated mastiff dog that died unexpectedly with locally extensive, acute, necrotizing myocarditis and myocardial infarction. intralesional protozoal tachyzoites in the affected myocardium were confirmed to be neospora caninum by a novel multiplex polymerase chain reaction (pcr) and immunohistochemistry. protozoal organisms were not identified in other tissues by histology, immunohistochemistry, or pcr. the multiplex pcr assay was used to quickly provide ...200516475515
in vitro isolation and characterisation of the first canine neospora caninum isolate in australia.neospora caninum was isolated and established in vitro from the skin lesion of a naturally infected dog. the identity of the parasite was evaluated by immunofluorescent antibody test (ifat), microscopy, western blotting and polymerase chain reaction (pcr). n. caninum dna was detected in the whole blood, serum, skin lesion, rectal scrapings and faeces of the infected dog utilising a nested pcr targeting the nc-5 gene of n. caninum. antigenic and genetic characterisation of the isolate, designated ...200616487658
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