Phenotypic characterisation of the cellular immune infiltrate in placentas of cattle following experimental inoculation with Neospora caninum in late gestation

Despite Neospora caninum being a major cause of bovine abortion worldwide, its pathogenesis is not completely understood. Neospora infection stimulates host cell-mediated immune responses, which may be responsible for the placental damage leading to abortion. The aim of the current study was to characterize the placental immune response following an experimental inoculation of pregnant cattle with N. caninum tachyzoites at day 210 of gestation. Cows were culled at 14, 28, 42 and 56 days post inoculation (dpi). Placentomes were examined by immunohistochemistry using antibodies against macrophages, T-cell subsets (CD4, CD8 and γδ), NK cells and B cells. Macrophages were detected mainly at 14 days post inoculation. Inflammation was generally mild and mainly characterized by CD3⁺, CD4⁺ and γδ T-cells; whereas CD8⁺ and NK cells were less numerous. The immune cell repertoire observed in this study was similar to those seen in pregnant cattle challenged with N. caninum at early gestation. However, cellular infiltrates were less severe than those seen during first trimester Neospora infections. This may explain the milder clinical outcome observed when animals are infected late in gestation.     

Neospora caninum tachyzoites inoculated by the conjunctival route are not vertically transmitted in pregnant cattle: A descriptive study

The aim of this study was to evaluate whether Neospora caninum tachyzoites (Nc-1) inoculated by the conjunctival route in pregnant cows were able to generate infection in their fetuses. Group 1 contained 2 naturally infected cows; group 2 contained two cows inoculated intravenously with 2.5 × 10⁸ tachyzoites, group 3 contained two cows inoculated with 2.5 × 10⁸ tachyzoites by the conjunctival route, and group 4 contained two uninfected control cows. The four inoculated cows from groups 2 and 3 were challenged at 23 weeks of gestation. An indirect fluorescent antibody test (IFAT), recombinant NcGRA7-based ELISA, ELISA for IgG subisotypes and Western blot analysis were assessed to characterize the humoral immune response in dams. Sera from their fetuses were tested also using Western blot analysis. Routine microscopic evaluation of H&E stained fetal tissues was made and any fetal tissues and placentas with lesions compatible with Neospora-infection were processed by immunohistochemistry (IHC). DNA extraction from fresh and formalin-fixed, paraffin-embedded fetal tissues were tested by nested PCR. All dams from groups 1, 2 and 3 were seropositive by IFAT, rNcGRA7-based-ELISA and Western blot. IgG₁/IgG₂ ratios were ≤1 at weeks 27 and 29 of gestation. Only fetuses from groups 1 and 2 developed N. caninum specific antibodies by Western blot. Histopathological lesions compatible with those caused by N. caninum were observed in fetuses from groups 1 and 2. N. caninum cysts and tachyzoites were observed by IHC on fetal tissues from groups 1 and 2. Only fetal samples from group 2 were positive by PCR. Further work is needed not only to characterize the cellular immune response but also to clarify the consequences on the dam after conjunctival inoculation of N. caninum tachyzoites. This study shows that N. caninum tachyzoites inoculated by the conjunctival route were not vertically transmitted in pregnant cows.     

Effect of vaccination of cattle with the low virulence Nc-Spain 1H isolate of Neospora caninum against a heterologous challenge in early and mid-gestation

Live vaccines have emerged as one of the most potentially cost-effective measures for the control of bovine neosporosis. Previous studies have shown that Nc-Spain 1H is a naturally attenuated isolate of Neospora caninum and that immunisation with live Nc-Spain 1H tachyzoites generated a protective immune response in mice. The aim of this study was to evaluate the safety and efficacy of immunisation in cattle. N. caninum-seronegative heifers were immunised subcutaneously twice with 10⁷ live Nc-Spain 1H tachyzoites prior to artificial insemination. No adverse reactions or negative effects on reproductive parameters were recorded following immunisation. In immunised and non-challenged heifers, no foetal deaths were observed, and none of the calves was congenitally infected. The efficacy against N. caninum-associated foetal death and vertical transmission was determined after challenge with high doses of the Nc-1 isolate at 70 and 135 days of gestation, respectively. After the challenge in early gestation, the immunisation induced a protection of 50% against foetal death. In addition, the microsatellite analysis performed in PCR-positive tissue samples from foetuses that died after challenge infection showed that the profiles corresponded to the challenge isolate Nc-1. A degree of protection against vertical transmission was observed after challenge at mid-gestation; calves from immunised heifers showed significantly lower pre-colostral Neospora-specific antibody titres than calves from the non-immunised/challenge group (P < 0.05). Strong antibody and interferon gamma responses were induced in the immunised heifers. This study indicates that the immunisation before pregnancy with the Nc-Spain 1H vaccine isolate appeared to be safe and reduced the occurrence of N. caninum-associated abortion and vertical transmission in experimentally infected cattle. In light of these encouraging results, the next step for testing this live attenuated candidate should be the assessment of its efficacy and safety in naturally infected cattle.     

In vivo cross-reactivity of Taenia saginata and Taenia crassiceps antigens in bovine cysticercosis

Steers sensitized or infected with Taenia saginata exhibited similar delayed-type dermal hypersensitivity (DTH) responses after intradermal inoculation with T. saginata or T. crassiceps skin test antigens. Steers sensitized to T. crassiceps cysticerci exhibited similar DTH responses to intradermal inoculation with T. crassiceps, T. saginata whole worm and T. saginata cysticerci antigens. No correlation existed between the DTH responses and the number of cysticerci in the carcasses. One sensitized/infected and one infected steer harbored cysticerci but exhibited no DTH responses. Infection with cysticerci did not elevate DTH responsiveness in sensitized animals.     

Experimental infection with a low virulence isolate of Neospora caninum at 70 days gestation in cattle did not result in foetopathy

The Nc-Spain 1H isolate of Neospora caninum, which was newly obtained from the brain of a congenitally asymptomatic infected calf, demonstrated a reduced in vitro tachyzoite yield and viability rate, as well as low virulence in mouse models. The objective of the present study was to determine the ability of this isolate to induce foetal death in a pregnant bovine model. For this purpose, 13 naïve pregnant heifers were divided into three groups and were experimentally challenged with either 10⁷ tachyzoites of Nc-1 (group 1, n = 5), Nc-Spain 1H (group 2, n = 5) isolates or phosphate-buffered saline (group 3, n = 3) intravenously at 70 days of gestation. After inoculation, pregnancy was monitored and dams were sacrificed when foetal death was detected. The remaining animals were slaughtered at 45 days post-infection. Maternal and foetal samples were collected for examination by histology and parasite DNA detection. Parasitaemia, specific anti-N. caninum IgG and interferon γ responses were also studied. At 3–4 weeks after infection, foetal death was detected in 3 out of 5 Nc-1-infected dams. However, no evidence of foetal death was observed in either Nc-Spain 1H-infected or control groups during the period studied. The most severe histopathological lesions were observed in the placenta and foetal organs from Nc-1-infected cattle that exhibited foetal death. It was in these animals that N. caninum DNA was more frequently detected. Parasitaemia was observed in all Nc-1-infected dams and in only 3 out of 5 Nc-Spain 1H-infected animals. The magnitude of the immune response was significantly higher in the Nc-1-inoculated group than in the group inoculated with the Nc-Spain 1H isolate. These data reveal the reduced virulence of the Nc-Spain 1H isolate in cattle.     

Evaluation of bovine cutaneous delayed-type hypersensitivity (DTH) to various test antigens and a mitogen using several adjuvants

The Bacillus Calmette Guerin (BCG)-induced/purified protein derivative (PPD)-elicited tuberculin skin test is a reliable measure of cell-mediated immune response (CMIR), specifically delayed-type hypersensitivity (DTH); however, its use in livestock may confound diagnosis of Mycobacterium tuberculosis. Therefore, various alternative antigen/adjuvant combinations were evaluated as inducers of DTH that were compared to the BCG/PPD test system with the purpose of finding a skin DTH protocol that does not cross-react with the tuberculin test and allows identification of high and low CMIR responder phenotypes. Specifically, 30 non-lactating cows (five/treatment) were sensitized on day 0 with mycobacteria [BCG, M. tuberculosis or Mycobacterium phlei cell wall extract (MCWE)], and ovalbumin (OVA) emulsified in Freund's complete adjuvant (FCA), non-ulcerative Freund's adjuvant (NUFA), complete NUFA or MCWE. On day 21, cows were injected intradermally with various test antigens including PPD tuberculin, phlein, and OVA. Phosphate buffered saline was included as the negative control and the T-cell mitogen phytohemagglutinin (PHA) was also administered. Double skin-fold thickness was evaluated before and at 6, 24, and 48 h post-injection. Skin biopsies were taken at 24 and 48 h to assess oedema, necrosis, and inflammatory cell infiltration. BCG/PPD and M. phlei/phlein treatments when given with a Freund's adjuvant induced equivalent DTH with peak reactions at 24-48 h after antigen injection. Cows receiving NUFA had fewer injection site granulomas than FCA or CNUFA treatments. The change in skin thickness response to PHA peaked at 6 h. Only cows receiving mycobacteria in NUFA had skin response to OVA, which peaked 6-24 h post-injection. Only sites tested with PPD or phlein had significantly higher lymphocyte infiltration than control, whereas neutrophils were significantly higher at PHA test sites and eosinophils predominated at the PHA test sites. Macrophages were significantly more numerous at the PPD and/or phlein test sites in treatment groups that received killed mycobacteria in a Freund's adjuvant and/or with BCG, and at the PHA test sites in all treatment groups. It was concluded that the M. phlei/phlein system did induce DTH and was similar to the DTH induced by the BCG/PPD system when MCWE was administered with a Freund's adjuvant. Therefore, this protocol is suitable for detecting high/low CMIR responders in research herds. However, cross-reaction to PPD was evident following induction of DTH using M. phlei. Hence, this protocol does not alleviate the problem of artificial induction of DTH cross-reactivity and would not be suitable for commercial herds where tuberculin testing is required.     

Isolation and molecular characterisation of Neospora caninum in cattle in New Zealand

AIM: To isolate Neospora caninum from the brains of naturally infected cattle and use molecular techniques to characterise the isolates.

METHODS: Neospora caninum tachyzoites were isolated in Vero cell culture from the brains of a cow and two calves. The isolates were characterised using polymerase chain reaction (PCR) methods, DNA sequencing, an immunofluorescent anti-body test (IFAT), transmission electron microscopy (TEM), and immunohistochemistry (IHC). The brains of the three cattle were subjected to histopathological examination. A pathogenicity study was conducted in 120 BALB/c mice.

RESULTS: Neospora caninum tachyzoites were isolated from all three cases and first observed in vitro between 14 and 17 days post-inoculation. Parasites were sub-cultured and maintained in Vero cell culture for more than 6 months. PCR products were generated for all three isolates, using two different primers. Sequencing of the PCR products and a subsequent BLAST search identified the isolates as N. caninum. In addition, the isolates tested positive using IFAT and IHC, and ultrastructure revealed by TEM was characteristic of N. caninum. Histopathological examination revealed lesions characteristic of N. caninum in 1/3 brains. In the pathogenicity study using BALB/c mice, the mortality rate was 3–7%.

CONCLUSION: This was the first successful isolation of N. caninum in New Zealand confirmed using molecular characterisation tests.     

Neospora caninum infection during early pregnancy in cattle: how the isolate influences infection dynamics,clinical outcome and peripheral and local immune responses

This work studies the influence of Neospora caninum intra-species diversity on abortion outcome, infection dynamics in terms of parasite dissemination and peripheral-local immune responses in pregnant cattle. Animals were intravenously inoculated at day 70 of pregnancy with 10⁷ tachyzoites of two isolates showing marked differences in virulence in vitro and in pregnant mouse models: Nc-Spain7, a high virulence isolate, and Nc-Spain8, a low-to-moderate virulence isolate. After inoculation, pregnancy was monitored, and dams were culled when foetal death was detected. Foetal mortality occurred in all infected heifers between days 24 and 49 post-infection (pi), however, it was detected sooner in Nc-Spain7-infected animals (median day = 34) than those inoculated with Nc-Spain8 (median day = 41) with a trend towards significance (P < 0.11). Similar histological lesions were observed in placentomes and in most of the foetuses from the two infected groups. However, parasites were more frequently detected in the placenta and foetuses by PCR and in the foetal brain by immunohistochemistry in Nc-Spain7-infected animals. Specific antibodies were detected starting at day 13 post-infection in all infected cattle, with higher IgG levels in Nc-Spain7-infected group. IFN-γ and IL-4 profiles also varied between infected groups in PBMC stimulation assays. Infected animals showed significant increases in their cytokine mRNA levels (IFN-γ, IL-4, IL-10, IL-12p40 and TNF-α) in the caruncle at time of foetal death. Differences between the infected groups were also observed for cytokine profiles. These results demonstrate the influence of the N. caninum isolate on foetal death outcome, infection dynamics and immune responses in cattle.     

Serological survey of Neospora caninum and Toxoplasma gondii in cattle (Bos indicus) and water buffaloes (Bubalus bubalis) in ten provinces of Brazil

The objective of this study was to determine the prevalence of antibodies to Neospora caninum and Toxoplasma gondii among 500 cattle (Bos indicus) and 500 buffaloes (Bubalus bubalis) using the indirect fluorescent antibody test (IFAT) technique. Blood samples from were collected from water buffalo and cattle in 10 municipalities in the northern region of Brazil. The frequency of cattle and water buffaloes seropositive for Neospora caninum in Pará state, Brazil, was 55% and 44%, respectively, and the frequency of cattle and water buffaloes seropositive for Toxoplasma gondii was 52% and 39%, respectively. Seropositivity for both N. caninum and T. gondii was detected in 10.6% of the cattle samples and 14.8% of the buffalo samples. The frequency of cattle positive for N. caninum and T. gondii was significantly (p < 0.05) higher than that of buffalo in two and three provinces, respectively. Buffaloes had a lower seroprevalence for N. caninum or T. gondii in all of the provinces studied. These results suggest that both species, when exposed to the same risks for N. caninum and T. gondii infection, have a high serological prevalence. Cattle showed a higher probability of being seropositive when exposed to the same risks for N. caninum and T. gondii. Our study, which included an extensive number of blood samples, provides important epidemiological information pertinent to buffalo production in tropical countries that can be used as a basis for disease-management practices in Latin America.     

Neospora caninum associated with epidemic abortions in dairy cattle: The first clinical neosporosis report in Turkey

Neospora caninum, a protozoan parasite, has been considered as one of the most important etiological agents responsible for abortion in dairy cattle throughout the world since it was first identified in dogs in 1988. In this report, characteristics of neosporosis, detected in a dairy cow ranch having epidemic abortions as high as 18.4%, were described. Blood samples were collected from 25 infertile or aborted dairy cattle, 6 calves born in 2006 and 40 heifers that were born in 2005 and raised in the same ranch. Necropsy was conducted in a 20-day-old Simmental calf that exhibited neurological signs including incoordination, head shaking, hyperextension in forelimbs and hindlimbs and tremor. The seroprevalance in aborted or infertile dairy cattle, heifers, and calves was 60%, 40%, and 33.3%, respectively. The mothers of seropositive two calves including clinically affected calf and its dam were N. caninum seropositive. In immunoperoxidase examinations, N. caninum antigen immunopositivity was observed in the degenerative and necrotic neurons in the brain, cerebellum as well as neurons in dorsal root ganglia of the cervical and thoracic regions of the spinal cord. In the heart, myocytes and Purkinje cells exhibited granular and linear patterns of immunoreactivity. Striated myofibers around the eyeball also showed immunolocalization for N. caninum antigen. Ultrastructurally, tachyzoites with typical apical complex, rhoptries and double-layered parasitic membrane were detected in the brain and heart sections. In conclusion, this report described clinical neosporosis for the first time in Turkey with tissue localization of the causative agents. This scientific communication also discusses the possible impact of cattle neosporosis by clinical, serologic and pathologic evidences collected from the survey of calves born in two successive generations in a ranch.     

Seroprevalence of antibody to NcSAG1 antigen of Neospora caninum in cattle from Western Java,Indonesia

Neospora caninum can cause fetal abortion and neonatal mortality in cattle, and is a cause of economic concern worldwide. This study aimed to determine the prevalence of Neospora caninum-specific antibodies in cattle from Western Java, Indonesia. Serum samples from 991 cattle from 21 locations were tested for antibodies to N. caninum by using an enzyme-linked immunosorbent assay (ELISA) on the basis of recombinant NcSAG1. The overall seroprevalence was 16.6%, ranging from 0 to 87.5% in the sampled locations. The results of this study indicate latent infection rates of sampled animals were different in each location. Further studies are necessary to elucidate the relationship between N. caninum infection and abortion in cattle, and to identify risk factors for infection in high-prevalence environments.     

Myeloid differentiation factor 88 is required for resistance to Neospora caninum infection

Neospora caninum is an intracellular parasite that causes major economic impact on cattle raising farms, and infects a wide range of warm-blooded hosts worldwide. Innate immune mechanisms that lead to protection against this parasite are still unknown. In order to investigate whether myeloid differentiation factor 88 (MyD88) is required for resistance against N. caninum, genetically deficient mice (MyD88^−/−) and wild type littermates were infected with live tachyzoites and the resistance to infection was evaluated. We found that sub-lethal tachyzoite doses induced acute mortality of MyD88^−/− mice, which succumbed to infection due to uncontrolled parasite replication. Higher parasitism in MyD88^−/− mice was associated with the lack of IL-12 production by dendritic cells, delayed IFN-γ responses by NKT, CD4⁺ and CD8⁺ T lymphocytes, and production of high levels of IL-10. MyD88^−/− mice replenished with IL-12 and IFN-γ abolished susceptibility as the animals survived throughout the experimental period. We conclude that protective IFN-γ-mediated immunity to N. caninum is dependent on initial MyD88 signaling, in a mechanism triggered by production of IL-12 by dendritic cells. Further knowledge on Toll-like receptor recognition of N. caninum antigens is encouraged, since it could generate new prophylactic and therapeutic tools to control parasite burden.     

Adaptive immune response, survival, and somatic cell score between postpartum Holstein and Norwegian Red x Holstein first-calf heifers

The objectives of this study were to evaluate antibody (AMIR) and cell-mediated immune responses (CMIR), survival, and somatic cell score (SCS) between purebred Holstein (HO) and crossbred Norwegian Red × Holstein (NRHO) first-calf heifers postpartum. Additionally, immune response traits observed as calves in a previous study were correlated with their immune response traits as first-calf heifers. Heifers, previously immunized as calves, were bled and reimmunized 6 to 9 d postcalving with known type 1 and type 2 antigens and human serum albumin (HSA). Seven days later, heifers were rebled, and background skinfold measurements of the tail fold were taken. Intradermal injections of PBS and type 1 antigen were administered on either side of the tail fold. On d 9 final skinfold measurements were taken and used to assess delayed-type hypersensitivity (DTH) as an indicator of CMIR. Blood samples were also collected for a final time on d 14 from heifers that received the antigen HSA. Serum was obtained from blood collected on d 0, 7, and 14 and analyzed by ELISA to assess AMIR. Data on survival and somatic cell count, which was converted to SCS, were obtained from CanWest Dairy Herd Improvement (DHI). All SCS, survival, and immune response data were analyzed using general linear models to determine significance between HO and NRHO first-calf heifers. To determine residual correlations between immune response traits observed in calves to their responses as first-calf heifers, residuals were obtained from models, and correlations between traits were determined using PROC CORR in SAS. Results showed NRHO had a greater primary IgG antibody response to HSA and greater tertiary IgG antibody response to the type 2 antigen compared with HO. Crossbreds (NRHO)also had significantly greater DTH response (P < 0.05) and, in general, greater survival from calving to 100 d in milk (dim), 100 to 305 dim, calving to 305 dim, and age at immune response testing as calf to 305 dim. No difference was observed between breeds for SCS. Results also showed most correlations between calf and first-calf heifer immune response traits were found to be positive and significant (P < 0.05). In conclusion, NRHO heifers have greater survival, which likely relates at least in part to increases in aspects of both AMIR and CMIR and could indicate that crossbred heifers have enhanced disease resistance.     

Canine Cutaneous Neosporosis: Clinical Improvement with Clindamycin

Résumé— Une dermite pyogranulomateuse provoquée par le protozoaire Neospora caninum a été diagnostiquée chez un Golden Retriever de 12 ans. Le nombreux nodules fistulisés étaient localisés au niveau de la tête et du thorax. De nombreux tachyzoites de N. caninum ont été retrouvés dans les biopsies des nodules et le diagnostic a été confirmé pas immunohistologie et examen en microscopie électronique. Le chien avait un titre de sérologie Neospora caninum de 3200 par immunofluorescence indirecte. Après un traitement à base de clindamycine pendant 45 jours, les lésions cutanées ont disparu. Le chien est probablement mort à cause d'un lymphosarcome. Il existait aussi une infection latente àToxoplasma gondii. les Neospora caninum n'ont pas pu être retrouvés par des techniques biologiques ni en culture ou par inoculation de souris à partir de prélèvements nécropsiques. Seuls des tachyzoites dégenérés ont pu être observés histologiquement. Ces observations montrent que la néosporose peut être envisagée dans le diagnostic différentiel des dermites pyogranulomateuses du chien et que la clindamycine est un médicament efficace pour traiter la néosporoe canine. [Dubey, J. P., Metzger, F. L., Hattel, A. L., Lindsay, D. S., Fritz, D. L. Canine cutaneous neosporosis: clinical improvement with clindamycin (Néosporose cutanée canine: amélioration clinique par la clindamycine). Resumen— Se diagnosticó una dermatitis piogranulomatosa causada por el protozoo parásito Neospora caninum en un perro de raza Golden Retriever de 12 años. El animal presentaba varios nódulos en la piel de la cabeza y tórax. Se observaron numerosos taquizoitos de N. caninum en los cortes histológicos de tejido obtenido mediante biopsia de dichos nódulos y el diagnóstico fue confirmado por tinción inmunohistológica y por microscopia electrónica. El perro mostró un titulo de anticuerpos contra N. caninum de 1:3,200 en la prueba de fluorescencia indirecta. Las lesiones cutáneas se resolvieron tras un tratamiento con hidroclorido de clindamicina durante 45 dias. El perro murió posteriormente a causa de un linfoma y presentaba también una infestación latente por Toxoplasma gondii. No se pudo demostrar la presencia de Neospora caninum mediante bioensayos en cultivos celulares ni en ratones inoculados con tejido canino obtenido en la necrospia. Tan solo se pudieron observar taquizoitos degenerados de N. caninum en tejido cutáneo obtenido en la necrospia. Estos hallazgos indican que se debe incluir neosporosis en el diagnóstico diferencial de dermatitis piogranulomatosas en el perro y que la clindamicina puede ser un fármaco eficaz para el tratamiento de la neosporosis cutánea. [Canine cutaneous neosporosis: clinical improvement with clindamycin (Neosporosis cutánea canina: mejora clinica con clindamicina). Abstract— Pyogranulomatous dermatitis caused by the protozoan parasite Neospora caninum was diagnosed in a 12-year-old Golden Retriever dog. Multiple draining nodules were located in the skin of the head and thorax. Numerous tachyzoites of N. caninum were found in histologic sections of the biopsy tissue from the cutaneous nodules and the diagnosis was confirmed by immunohistochemieal staining and by electron microscopic examination. The dog had a 1:3200 serum antibody titer to N. caninum in the indirect fluorescent antibody test. The cutaneous lesions resolved after a 45-day treatment with clindamycin hydrochloride. The dog eventually died because of lymphosarcoma and also had a latent infection with Toxoplasma gondii. Neospora caninum could not be demonstrated by bioassays in cell culture or mice inoculated with canine tissue obtained at necropsy. Only degenerating N. caninum tachyzoites were seen in skin tissue taken at necropsy. These observations indicate that neosporosis should be considered in the differential diagnosis of pyogranulomatous dermatitis in dogs and that clindamycin may be an effective drug for treating cutaneous neosporosis.     

Prevalence and risk factors associated with Neospora caninum infection in dairy herds in Jordan

A cross-sectional study was carried out to determine the seroprevalence and to identify risk factors associated with Neospora caninum infection in 62 dairy herds (n?=?671 cows) in Jordan between January and June 2007. Information regarding herd management was recorded through personal interviews with farmers. Antibodies against N. caninum were detected using an indirect ELISA test. Chi-square analysis and multivariable logistic regression model were used to identify risk factors associated with N. caninum seropositivity. The true prevalence of antibodies against N. caninum in individual cows and cattle herds was 35 and 66.5 %, respectively. There was no significant difference in the percentage of seropositive animals between different age groups. Multivariable logistic regression model revealed workers frequently visiting nearby farms as a risk factor for seropositivity to N. caninum, while presence of a calving pen was suggested as a protective factor. Amman, Balqa, and Mafrak governorates had significantly lower seroprevalence to N. caninum compared to other Jordanian governorates. Results of this study indicated that N. caninum infection may be widespread in Jordan.     

Neospora caninum and complex vertebral malformation as possible causes of bovine fetal mummification

Bovine neosporosis, caused by Neospora caninum is a leading cause of abortion in cattle. We postulated that neosporosis could lead to fetal death and mummification. Fifteen mummified fetuses were tested by polymerase chain reaction (PCR) for the mutation in the bovine SLC35A3 gene that causes complex vertebral malformation (CVM) and the pNC-5 gene which identifies N. caninum infection. DNA was extracted from the mummified fetuses and the sex of the mummies was determined by PCR. The CVM mutation was not detected in the mummified fetuses, but 4 fetuses were positive for N. caninum infection. The ages of the mummies with N. caninum infection were 100, 113, 123, and 131 days. Twelve of the 15 mummified fetuses were male. To our knowledge, this is the first detection of N. caninum as a possible cause of bovine fetal mummification.     

Neospora caninum versus Brucella spp. exposure among dairy cattle in Ethiopia: A case control study

This case-control study aimed at assessing the relative association of Neospora caninum and Brucella species exposure with reproductive disorders. The study was carried out between October 2011 and June 2012 on 731 dairy cows sampled from 150 dairy farms in selected 17 conurbations of Ethiopia. Two hundred sixty-six of the cows were categorized as cases based on their history of abortion or stillbirth while the remaining 465 were controls. The presence of antibody to N. caninum was screened using indirect ELISA, while Brucella spp. exposure was assayed serially using Rose Bengal Plate Test and Complement Fixation Test. Exposure to N. caninum was more frequently observed among cases (23.8 %) than controls (12.7 %), while no significant difference (p?>?0.05) was noted for Brucella exposure between the two groups. Moreover, the proportion of cows with disorders like retention of fetal membrane, endometritis and increased inter-calving period were significantly higher (p?<?0.05) among Neospora seropositive cows. In conclusion, the finding discloses the strong association of N. caninum with reproductive disorders compared to Brucella spp. exposure. However, neither N. caninum nor Brucella spp. could explain the majority (73.2 %) of the reported abortions and stillbirths in cattle. Hence, this observation underscores the need for more intensive investigation on the identification of causes of the aforementioned disorders in dairy cattle of Ethiopia.     

Determination and correlation of anti-Neospora caninum antibodies in dogs and cattle from Mexico

The aim of the present study was to determine and to compare through an indirect enzyme linked immunosorbent assay (ELISA) test, the presence of anti-Neospora caninum antibodies in city and farm dogs, as well as in farm cows, and the relationship among them. The correlation between anti-N. caninum antibodies in farm dogs and cattle was also assessed. The research was conducted in the dairy region of Tizayuca, Hidalgo, Mexico. The frequency of anti-N. caninum antibodies was significantly higher in farm dogs (n = 14) (51%) when compared to those from the city (n = 6) (20%) (P < 0.05), suggesting that farm dogs have a higher risk of exposure to the parasite. There was no significant difference in seropositivity between males (n = 11) (39%) and females (n = 9) (33%) (P > 0.05). The frequency of anti-N. caninum antibodies in farm cattle was significantly higher in farms with dogs (n = 158) (58%) when compared to those with no dogs (n = 43) (35%) (P < 0.05). These results suggest the possible transmission of the parasite from dogs to cattle.     

Type 1 and type 2 immune response profiles of commercial dairy cows in 4 regions across Canada

Diseases of dairy cattle have adverse implications for both the dairy industry and animal welfare. Understanding adaptive immune response profiles of cattle on a national scale will provide insight into the potential for improving health and decreasing disease. The objectives of the present study were to evaluate immune response phenotypes of Holstein cows outside the peripartum period and to determine if antibody isotype bias to putative type 1 and type 2 test antigens is maintained. The cows, housed on commercial farms in 4 key dairy regions across Canada, were immunized with test antigens to measure their ability to mount cell-mediated immune responses (CMIR) and antibody-mediated immune responses (AMIR). Delayed-type hypersensitivity (DTH) was used as an indicator of CMIR and primary and secondary serum antibodies of the immunoglobulin (Ig) G1 and IgG2 isotypes were used to determine AMIR to the test antigens. Immune response phenotypes varied significantly among regions, herds, and cows. Cows in Alberta had significantly higher DTH responses and secondary responses to the type 2 test antigen than those in other regions. However, cows in Alberta had significantly lower primary antibody responses. It was found that Alberta had the lowest incidence of mastitis caused by Escherichia coli and Staphylococcus aureus compared with other regions. The IgG1/IgG2 antibody isotype ratio confirmed the nature of the test antigens. This was the first study to evaluate adaptive immune response profiles and disease incidence of dairy cows on a national scale and it therefore provides a glimpse of the current situation in Canada.     

Immunoglobulin isotypes of lactating Holstein cows classified as high, average, and low type-1 or -2 immune responders

Infectious diseases are detrimental to the health and economy of the livestock industry. Observations of cattle resistant to natural infections have implied the feasibility of breeding livestock for disease resistance. Studies of pigs selected for antibody (AMIR)- and cell (CMIR)-mediated immune responses have demonstrated increased immune responsiveness suggesting enhanced protection by both type 2 and type 1 responses, respectively. Additionally, natural or artificial infections of cattle suggest that the production of particular immunoglobulin (Ig) M, IgG1 and IgG2 isotypes are important for protecting against pathogens. In fact, IgG1/IgG2 ratios are often used to establish whether type 1 (CMIR) or type 2 (AMIR) responses predominate following immunization or infection. The objectives of this study were therefore; (1) to evaluate the Ig isotype bias responses to Candida albicans and hen-egg white lysozyme (HEWL) in cows classified as high responders (HR), average responders (AR) or low responders (LR) based on AMIR or CMIR; (2) to determine if ranking based on IFN-γ (a type 1 cytokine) and DTH responses were analogous in terms of ranking; and (3) to estimate IFN-γ, Ig isotypes, and DTH correlations. Antibody responses to HEWL and DTH to C. albicans were detected such that cows were phenotypically classified as HR, AR and LR for AMIR or CMIR with significant differences (p ≤ 0.05) among classified groups. C. albicans-induced IFN-γ allowed classification of cows, some of which had the same ranking as that of DTH response. The lowest IgG1/IgG2 ratio was to the C. albicans purified antigen (candin), but no differences were observed in anti-HEWL or anti-candin IgG1/IgG2 ratios between classified groups. Anti-HEWL IgG1 and IgG2 responses at day 21 post-immunization were negatively and significantly correlated with DTH to candin at 24h. There were no significant correlations between anti-HEWL or anti-candin IgG1 or IgG2 responses with IFN-γ. Based on Ig isotype bias, IFN-γ and DTH responses, it was concluded that immunization with C. albicans can be used to classify CMIR responder cows based on DTH read-out.