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Seroprevalence of antibodies against bovine leukemia virus, bovine viral diarrhea virus, Mycobacterium avium subspecies paratuberculosis, and Neospora caninum in dairy cattle in Saskatchewan.

Blood was drawn from 1530 dairy cows in 51 herds. For antibodies against bovine leukemia virus, Mycobacterium avium subspecies paratuberculosis, and Neospora caninum, 37.4%, 2.7%, and 5.6% of cows were test positive, respectively, while 29.2% of herds had unvaccinated animals with > or = 1:64 for bovine viral diarrhea virus.

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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 prevalence of the infection in Canadian dairy and beef cattle are reviewed and briefly compared with estimates from other parts of the world. Most importantly for Canadian bovine practitioners, the impacts of the infection, risk factors for its occurrence, and methods of control are also discussed. By reviewing the scientific literature on N. caninum from a Canadian perspective, culling decisions based on the interpretation of diagnostic tests are more effectively made in the control of N. caninum-associated disease.

Abortion, Veterinary↗

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, it appeared that different immunoglobulin classes recognizing different antigenic molecules were cooperating to cope with neosporosis.

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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 glycoproteins in the pathogenesis of Neospora-associated myositis is discussed.

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Molecular identity of the electrophoretic pattern and the major cross-reacting immunogens of Toxoplasma gondii (RH strain) and Neospora caninum.

The complex electrophoretic profile of the antigenic components of Toxoplasma gondii and Neospora caninum was studied. T. gondii (RH strain) trophozoite's antigens were resolved by gel electrophoresis under denaturating condition. Resolved polypeptides were tested by immunoblotting against hyper-immune serum prepared in Boscat rabbits. Immunoblot analysis revealed 6 reactive bands, which are 158, 111, 102, 86, 55 and 33 kDa, out of 10 antigenic bands of the parasite. Concerning N. caninum trophozoite's antigens, immunoblot analysis revealed 5 reactive bands, which are 159, 119, 106, 97 & 39 kDa, out of 8 antigenic bands of the parasite. Cross reactivity trials between the 2 parasites explore 3 cross-reactive bands, indicating high antigenic similarity in between.

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Neospora caninum and Leptospira serovar serostatus in dairy cattle in Ontario.

No significant association existed between Neospora caninum titer and serostatus to Leptospira serovar hardjo, icterohaemorrhagiae, or pomona in cattle on 78 dairy herds in Ontario. Leptospira titer increased with parity. Amongst herds not vaccinated against Leptospira, the proportions of herds with > or = 1 animal seropositive to serovar hardjo, icterohaemorrhagiae, or pomona were 45%, 42%, and 58%, respectively.

Abortion, Veterinary↗

Seroprevalences of antibodies against bovine leukemia virus, bovine viral diarrhea virus, Mycobacterium avium subspecies paratuberculosis, and Neospora caninum in beef and dairy cattle in Manitoba.

Of 1204 dairy cows and 1425 beef cows sampled, 60.8% and 10.3% were seropositive for Bovine leukemia virus, 4.5% and 1.7% were seropositive for Mycobacterium avium subspecies paratuberculosis, and 8.3% and 9.1% were seropositive for Neospora caninum, respectively, while 28.1% of dairy herds had unvaccinated animals with titres > or = 1:64 for Bovine viral diarrhea virus.

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[Distribution of anti-Neospora caninum antibodies in dairy cows at municipalities of Resende and Rio Claro in the State of Rio de Janeiro, Brazil].

This study aimed to verify the prevalence of anti-Neospora caninum antibodies in dairy cows from municipalities of Resende and Rio Claro, both located at sulfluminense dairy belt. A total of 29 farms from Resende and 28 from Rio Claro were randomly selected. To evaluate the association between herds and animals against the etiological agent χ² tests were used. Seventy eight animals at Resende (25.74%, confidence intervals(IC) at 95%: 15.7-25.8%) had anti-N. caninum antibodies, while 53 animals (20.38%, IC 21-31.1%) at Rio Claro (p=0.16). No association between positive animals in relationship to municipalities was observed. When considered properties, 26 (89.7%, IC: 76, 3-94.9%) at Resende and 24 (85.71%, IC: 67.3-96%) at Rio Claro had at least a positive animal. This demonstrated that anti-N. caninum antibodies were observed among dairy cows at both municipalities. In spite of abortions were observed with relative frequency in endemic areas of N. caninum, they were not observed in the present study.

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[Reaction of bovine endothelial cells in vitro to coccidia (Eimeria bovis, Toxoplasma gondii, Neospora caninum) infections as the expression of a non-adaptive immune response].

Eimeria (E.) bovis sporozoites as well as Toxoplasma (T.) gondii and Neospora (N.) caninum tachyzoites can invade bovine endothelial cells (BUVEC) in vitro and develop to next stage meronts within 15-20 and 3-4 days, respectively. The latter differences suggest different immune evasion strategies, particularly concerning innate reactions. Realtime RT-PCR techniques were used to determine transcript levels of genes relevant in this sense, i.e. adhesion molecule, chemokine, growth factor GM-CSF, cyclooxygenase 2 (COX-2) and iNOS genes in infected cells. In addition, adhesion of neutrophils (PMN) to infected BUVEC monolayers was quantified. Effects differed between E. bovis and T. gondii/N. caninum as the latter two species strongly enhance the transcription of all genes in question and induce PMN adhesion to infected BUVEC whereas E. bovis either caused only weak responses or failed to enhance gene transcription as in case of CXC chemokines and COX-2. It even down regulates adhesion molecule expression in response to cytokines. The differences between the species may reflect differing immune evasion strategies, in case of E. bovis favouring its long lasting development to macromeronts.

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Seroprevalence of Mycobacterium avium subspecies paratuberculosis, Neospora caninum, Bovine leukemia virus, and Bovine viral diarrhea virus infection among dairy cattle and herds in Alberta and agroecological risk factors associated with seropositivity.

A province-wide, cross-sectional seroprevalence and agroecological risk factor study of Mycobacterium avium subspecies paratuberculosis (MAP), Neospora caninum (NC), Bovine leukemia virus (BLV), and Bovine viral diarrhea virus (BVDv) genotypes 1 and 2 (BVDv1 and BVDv2) infection in dairy cattle herds in Alberta was conducted. Among adults, the seroprevalence of MAP, NC, and BLV was 9.1%, 18.5%, and 26.9%, respectively. For MAP, based on a herd test cutpoint of 2 or more seropositive cows, 58.8% of herds were infected. Herd-level seroprevalence for NC and BLV was 98.7% and 86.7%, respectively, based on a herd-test cutpoint of 1 seropositive cow. Among unvaccinated dairy heifers, the seroprevalence for BVDv1 and BVDv2 infection was 28.4% and 8.9%, respectively, while herd-level infection was 53.4% and 19.7%. Seroprevalence for MAP varied moderately by agroecological region, whereas that for NC, BLV, and BVDv1 and BVDv2 did not. For MAP, aridity and soil pH (correlated features of the region) were also important.

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[Seroprevalence of Neospora caninum in cattle in the region of Kayseri].

This study was carried out in order to determine the seroprevalence of Neospora caninum in cattle in the region of Kayseri. Blood samples were collected from 186 cattle and sera were obtained from these samples. Sera samples were analyzed for antibodies against N. caninum using a commercially available competitive ELISA (c-ELISA) kit. According to the ELISA test results, seroprevalence of N. caninum in the region of Kayseri was found to be 7%. Three of 9 aborting cows (33.3%) were found to be seropositive. The seropositivity differences between aborting and non-aborting cows were statistically significant (p < 0.05).

Abortion, Veterinary↗

Distribution of antibodies against Neospora caninum, BVDV and BHV-1 among cows in brazilian dairy herds with reproductive disorders.

Neospora caninum, bovine viral diarrhea virus (BVDV) and bovine herpes virus 1 (BHV-1) are worldwide spread pathogens associated with reproductive problems in cattle. The present work aimed to observe the infection pattern of these three pathogens in two dairy herds with distinct reproductive managements from Triângulo Mineiro, Minas Gerais State, Brazil. The herds were not vaccinated against either N. caninum, BVDV or BHV-1. Blood samples were collected and analyzed for presence of specific antibodies, and N. caninum IgG avidity was measured in N. caninum positive samples. In herd 1, 34 out of 174 sampled cows (20%) had antibodies to N. caninum and the seropositivity of BVDV and BHV-1 were 62% and 86%, respectively. Of 69 sampled cows in herd 2. 7 (10%) had antibodies to N. caninum, and 49% and 39% were seropositive to BVDV and BHV-1, respectively. The IgG avidity profiles indicated that N. caninum had been present in both herds for some years and that herd 1 had an ongoing horizontal spread of the parasite. The results indicate that the studied reproductive pathogens were present in the herds and partly may have contributed to their reproductive problems.

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Vertical transmission of Neospora caninum in mice.

Herein we report a murine model to examine transplacental transmission, transmammary transmission, or both, of Neospora caninum. Prevalence of transplacental transmission in outbred Swiss-Webster mouse pups was 85%, with 11 of 13 litters containing at least 1 transplacentally infected pup. Sixty-two percent of litters born to experimentally infected dams contained 85% or more transplacentally infected pups. Numbers of pups congenitally infected per litter was higher if dams were inoculated during the first half of pregnancy. Transplacental transmission decreased to 25% when a singly infected dam delivered a second litter. Transmammary transmission was observed in 1 of 51 pups suckling dams experimentally infected 5, 10, or 15 days postparturition. No pups were infected when cross-fostered onto chronically infected dams.

Animals↗

Experimental oral inoculations in birds to evaluate potential definitive hosts of Neospora caninum.

Experimental oral inoculations to evaluate potential definitive hosts of Neospora caninum were conducted by feeding infected rodent tissues to 9 carnivorous birds of 4 species. Birds included 2 red-tailed hawks (Buteo jamaicensis), 2 turkey vultures (Cathartes aura), 2 barn owls (Tyto alba), and 3 American crows (Corvus brachyrhynchus). The rodents (mice or rats) had been inoculated with 100,000 culture-derived tachyzoites of N. caninum 1-6 mo before feeding to the birds. Fecal samples were collected from each bird daily for 1 mo after feeding rodents and examined for oocysts by fecal flotation. In addition, processed aliquots from all avian fecal samples were fed to BALB/c mice. Five weeks after feeding, mice were bled and sera were tested for antibodies against N. caninum. One to two months later, mice were killed and brain tissue was examined microscopically for protozoal cysts. While occasional oocysts were found in avian fecal samples, these were likely not N. caninum because they were not infective to BALB/c mice. It was concluded that the bird species tested are not likely to be definitive hosts of N. caninum.

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Mouse model for central nervous system Neospora caninum infections.

Neospora caninum is a protozoan parasite that causes severe disease in transplacentally infected dogs and abortions in domestic ruminants. Rodent models of neosporosis rely on treatment of hosts with methylprednisolone acetate (MPA) to enhance infections. The present study reports the development of an inbred BALB/c mouse model that results in central nervous system neosporosis in the absence of MPA treatment. Seven of 12 BALB/c mice died 26-70 days after subcutaneous (s.c.) inoculation with tachyzoites of the NC-1 strain of N. caninum, and none of 12 BALB/c mice died after s.c. inoculation with tachyzoites of the NC-3 strain. None of 8 HSD:ICR mice (4 mice, NC-1 strain; 4 mice, NC-3 strain) developed clinical neosporosis or died after s.c. inoculation with tachyzoites. Control BALB/c (2) and HSD:ICR (2) mice s.c. inoculated with Hanks' balanced salt solution did not develop clinical signs of disease. Some mice in all N. caninum-inoculated groups had brain lesions, but significantly (P < 0.05) more BALB/c mice inoculated with the NC-1 strain had brain lesions.

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Characterization of a murine monoclonal antibody generated against Neospora caninum tachyzoites by use of western blot analysis and immunoelectron microscopy.

A murine monoclonal antibody (MAB) 6G7 generated against tachyzoites of Neospora caninum recognized 8 major and several minor antigens, as observed by western blot analysis. Relative rate of migration of the 8 major antigens ranged from 31 to 97.4 kd. In addition, MAB 6G7 recognized a Toxoplasma gondii tachyzoite antigen with a relative rate of migration of 107 kd. Immunogold labeling of N caninum tachyzoites grown in human foreskin fibroblast cells indicated that MAB 6G7 binds to micronemes, dense granules, basal portions of rhoptries, and intravacuolar tubules within the parasitophorous vacuole. Monoclonal antibody 6G7 also bound to micronemes and basal portions of rhoptries within tachyzoites of T gondii. Monoclonal antibody 6G7 did not significantly inhibit development of tachyzoites in vitro.

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Examination of the activities of 43 chemotherapeutic agents against Neospora caninum tachyzoites in cultured cells.

Neospora caninum causes serious disease in dogs, and it, or a similar parasite, is a major cause of abortion in cattle. Little is known about the susceptibility of this protozoan to antimicrobial agents. We studied several antimicrobial agents to determine which classes might have activity against this parasite. We also determined whether activity of such agents was coccidiocidal or coccidiostatic. A 2-day of treatment, monoclonal antibody-based enzyme immunoassay and a 5-day of treatment, cell culture flask (CCF), lesion-based assay were developed to examine the ability of test agents to inhibit tachyzoite multiplication. Seven sulfonamides were examined, with the following activities observed: sulfathiazole > or = sulfamethoxazole > sulfadiazine > sulfaquinoxaline > or = sulfamethazine > sulfadimethoxine > sulfamerazine. Dapsone, a sulfone, had little activity. Six dihydrofolate reductase/thymidylate synthase inhibitors were examined, with the following activities observed: piritrexim > pyrimethamine > ormetoprim > trimethoprim = diaveridine > methotrexate. Six ionophorous antibiotics were examined; lasalocid, maduramicin, monensin, narasin, and salinomycin had equivalent activities, but alborixin was toxic for host cells at the lowest concentration examined. Three macrolide antibiotics--azithromycin, clarithromycin, and erythromycin--were examined and had equivalent activities. Two tetracycline antibiotics, doxycycline and minocycline, were examined and had equivalent activities. Three lincosamide antibiotics were examined, with the following activities observed: clindamycin hydrochloride > clindamycin phosphate > lincomycin hydrochloride. Pentamidine and 6 of its analogs were examined, and only hexamidine and 1,4-Di[4-(2-imidazolinyl)-2-methoxy-phenoxy]butane had activity. Eight miscellaneous antiprotozoal agents were examined for activity. Amprolium, metronidazole, paromomycin, and roxarsone had little activity.(ABSTRACT TRUNCATED AT 250 WORDS)

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Neospora caninum: specific oligonucleotide primers for the detection of brain "cyst" DNA of experimentally infected nude mice by the polymerase chain reaction (PCR).

Both tachyzoites and bradyzoites in tissue cysts of Neospora caninum are morphologically difficult to distinguish from those of other cyst-forming apicomplexan parasites such as Toxoplasma gondii. Thus, molecular tools may contribute to easy identification of the parasite. Based upon an N. caninum-specific DNA fragment that we have recently cloned, 5 sense (Np1, Np3, Np5, Np7, Np21) and 4 antisense (Np2, Np4, Np6, Np8) oligonucleotides were designed for a sensitive and specific polymerase chain reaction (PCR). Among 19 combinations of sense and antisense primers, the Np21/Np6, Np21/Np4, and Np7/Np6 primer pairs were found to generate specific single bands in the presence of at least 10 pg genomic parasite DNA as a template. The primer pair Np21/Np6 was able to detect a single tachyzoite in the background of DNA derived from 2 mg of brain tissue. In experimentally infected athymic ICR:nu/nu mice, N. caninum-DNA was detected consistently from brain tissue at days 13 and 18 after subcutaneous inoculation of tachyzoites. The presence or absence of the organisms in the cerebrum in either proliferative or cystic form was examined by immunohistological staining. The results indicate that PCR with the primer pair Np21/Np6 could provide an efficient tool for large-scale epidemiological studies using brain tissue obtained at necropsy.

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