Neospora abortions in dairy cattle: diagnosis, mode of transmission and control

AIM: To determine the contribution of Neospora caninum to abortions on a dairy farm in NSW (Australia), determine the mode of transmission and develop and trial a control option for infection. METHODS: Two whole herd bleeds were conducted 12 months apart and the association between serological status and abortion events were calculated for a number of bovine abortifacients. Family trees were constructed for N. caninum seropositive cattle in the herd. Some N. caninum seropositive cows were culled from the herd and no female offspring was retained from seropositive cows. RESULTS: At the first whole-herd bleed in December 2002 a seroprevalence of 10.2% for N. caninum infection was detected. Cows with N. caninum infection were 13 times more likely to abort than uninfected ones. Seventy-five percent of seropositive animals in the herd were related, suggesting a high degree of congenital infection/transmission. Only 15% of infections were likely to be postnatally acquired. Selective culling of seropositive cows and not breeding from them reduced the number of seropositive animals. Only one newly sero-converted cow was detected at the second whole-herd bleed 12 months later. CONCLUSIONS: Seroepidemiological approaches were able to establish a high degree of association between N. caninum infection and low-level abortion in the dairy herd. Vertical transmission of infection was the predominant mode of infection and hence control efforts aimed at selectively culling seropositive animals from the herd were highly successful in reducing the level of infection.     

Diagnosis and epidemiology of Neospora caninum-associated abortions in cattle]

Neospora caninum, a recently discovered protozoan parasite closely related to Toxoplasma gondii, has world-wide been recognized as an important cause of bovine abortion. N. caninum possesses a wide host range. The dog can be a definitive host for N. caninum. In cattle, the infection is transmitted transplacentally with high efficiency, while the majority of congenitally infected calves is clinically normal at birth and thereafter. Whether horizontal transmission occurs in cattle and whether this potential mode of transmission has epidemiological significance, remains to be elucidated. N. caninum-associated abortions can occur in epidemic or endemic form in a herd. The clinical symptoms of bovine neosporosis are confined to the occurrence of abortion, stillbirth and weak calves. Multifocal nonsuppurative encephalitis represents the most frequent pathohistological finding in N. caninum-associated abortions. The causative agent can be demonstrated by immunohistochemistry or polymerase chain reaction. Serological techniques can be used for indirect diagnosis. On the basis of the available diagnostic methods and the present knowledge about the epidemiology of the infection proposals are made regarding diagnosis, epidemiological assessment and prophylaxis of N. caninum-associated abortion problems in cattle herds.     

Economic considerations for diagnostic and control options for Neospora caninum-induced abortions in endemically infected herds of beef cattle

OBJECTIVE: To compare economic outcome for herds not exposed to Neospora caninum with that for herds with various seroprevalences of N caninum infection and evaluate 3 control strategies. DESIGN: Economic simulation model. SAMPLE POPULATION: Beef herds with various seroprevalences of N caninum infection. PROCEDURE: A 5-year simulation model was used. Control strategies that were evaluated included culling females that fail to calve, selling seropositive females and purchasing seronegative replacements, and excluding the daughters of seropositive dams as potential replacements. RESULTS: For a 5-year period with low prices for feeder calves, endemic N caninum infection decreased mean return to fixed assets by 22.2% when true seroprevalence was 10% and by 29.9% when true seroprevalence was 70%. Percentage decrease in return to fixed assets was less dramatic when a 5-year period with high prices for feeder calves was evaluated. Analysis indicated that 2 control strategies (culling females that fail to give birth to a calf and selling seropositive female cattle and purchasing seronegative replacement female cattle) were not likely to be economically beneficial. The third control strategy (testing the entire herd for N caninum infection and excluding the female offspring of seropositive dams as replacements) appeared to be a reasonable control strategy. CONCLUSIONS AND CLINICAL RELEVANCE: For the assumptions in the model, endemic N caninum infection decreases return to fixed assets for cow-calf herds. Of the potential control strategies evaluated, testing the entire herd for N caninum infection and excluding the daughters of seropositive dams as potential replacements provided the best economic return.)     

Comparison of serological methods for the diagnosis of Neospora caninum infection in cattle

The aims of this study were to evaluate the performance and agreement of various commercial and in-house Neospora caninum antibody assays used in dairy cattle in North America, and to investigate reproducibility of two assays performed in different laboratories. From 1998 to 2005, three enzyme linked immunosorbent assays (ELISAs, a competitive ELISA-VMRD Inc., an indirect ELISA-Biovet Inc., and another indirect ELISA-Herdchek IDEXX Corp.), two indirect fluorescent antibody tests (IFATs, VMRD Inc., and in-house USDA) and one N. caninum agglutination test (NAT, in-house USDA) were utilized to test 397 randomly selected dairy cattle serum samples from 34 herds in eastern Canada for antibodies to N. caninum. The manufacturers' recommended cut-off values were used to evaluate test performance and agreement between tests. One IFAT (VMRD Inc.) performed well (sensitivity and specificity: 0.97 and 0.97, respectively) using reference sera (n = 452), therefore, results from this IFAT on the 397 samples could subsequently be used as the reference standard to calculate test characteristics for the other assays. Only 11% of the 397 sera were found to be N. caninum-positive with the IFAT. Prevalence-adjusted bias-adjusted kappa (PABAK) ranged from 0.06 to 0.99. Positive agreement was moderate to very good (P(pos) = 0.25-0.96). Negative agreement was very good for all assays (P(neg) > 0.94) except NAT (P(neg) = 0.66). Sensitivity was > or =0.89 for all assays except the NAT, which had a significantly lower sensitivity (0.66). Specificity was high (>0.94) for all assays except for one indirect ELISA (specificity = 0.52). This indirect ELISA did not perform satisfactorily when used in 1998, but an improved version of the ELISA performed as one of the best assays in 2004. Reproducibility of the competitive ELISA was excellent, but the reproducibility of the indirect ELISA that was improved was low (concordance correlation coefficient = 0.90 and 0.36, respectively). The performance characteristics observed for most assays in this study make them useful for screening antibodies to N. caninum in cattle.     

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 Neospora caninum-like in two brains, were seen in 5% of aborted foetuses with lesions. No protozoa or associated lesions were seen in 57 late term foetuses or neonatal controls. A farm survey supported the hypothesis that Neospora is an important cause of multiple abortions in cattle. Laboratory and farm data suggested that protozoal abortion was more common in early gestation. No predisposing causes of abortion were found and there was no evidence of point infection of affected herds.     

Prevalence of Neospora caninum antibodies in dairy cattle and water buffaloes and associated abortions in the plateau of Southern Peninsular India

A seroprevalence study of bovine neosporosis was conducted among 1,927 dairy cattle and 341 water buffaloes from Karnataka and Andhra Pradesh states in plateau of southern peninsular India by employing competitive enzyme-linked immunosorbent assay. Overall, 12.61 and 9.97 % sera samples were found positive for the presence of Neospora caninum antibody, respectively, among cattle and water buffaloes. Out of 1,927 sera samples from cattle, 912 and 1,015 samples were collected from unorganized and organized herds, respectively. The cattle screened were of upgraded Holstein–Friesian and water buffaloes were of graded Surti breed. Significantly (p?<?0.05) higher prevalence was found in the cattle in unorganized herds (16.66 %) in comparison to organized herds (8.96 %). The highest seroprevalence was recorded in the age group of 4 years and above in both type of cattle herds and water buffaloes. There was a significant variation of seroprevalence (p?<?0.05) observed between different age groups of cattle. The rate of seroprevalence increased with the increment in the age of the animals suggesting a possibility of horizontal mode of transmission of the infection from the environment. The percentage of abortion history was more in seropositive group (51.65 %) in comparison to the seronegative group (5.84 %) and the seropositive cattle were 8.84 times more likely to experience abortion than the seronegative cattle. The occurrence of abortion among different age groups varied significantly (p?<?0.05). The findings revealed the presence of neosporosis in the southern peninsular India among cattle and water buffaloes and a strong association between the seroprevalence and abortion.     

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 abortions were attributed to N. caninum.     

Neosporosis-like abortions in a herd of dairy cattle

Twenty-nine cows from a 240-cow drylot Holstein dairy in north-central New Mexico aborted over a period of approximately 5 months. Nine aborted fetuses were necropsied, and all but 1 of the fetuses were between 5 and 7 months of gestation. Microscopic examination of fetal tissues revealed focal necrotizing encephalitis and nonsuppurative myocarditis in 7 of the 9 fetuses. Additional lesions observed in some fetuses were focal hepatic necrosis, nonsuppurative myositis, focal necrotizing placentitis, focal nonsuppurative pneumonia, and focal nonsuppurative nephritis. A few groups of Neospora caninum-like protozoan organisms were observed adjacent to necrotic foci in the brain of 2 fetuses and in the kidney of a third fetus. No other cause of abortion was detected.     

Potential involvement of Neospora caninum in naturally occurring ovine abortions in New Zealand

Neospora caninum is an obligate intracellular parasite and is recognised as the leading cause of bovine abortion worldwide. Natural infection with N. caninum has been described in sheep but it has generally not been regarded as a significant cause of abortion. Recently, there have been several New Zealand cases of foetal abortions where N. caninum was detected which strongly suggested the involvement of Neospora in these abortions. However, there is minimal information about the prevalence of N. caninum infection naturally occurring in New Zealand sheep flocks and particularly its impact on reproduction success. Thus, this present study provides preliminary data on the role that Neospora is playing in ovine reproductive failure by establishing the prevalence of N. caninum antibodies and DNA in ewe blood and foetal material present in 21 New Zealand sheep farms with ongoing unexplained abortion problems and 10 farms with consistently high fertility levels. The results of this study demonstrated an overall seroprevalence of 1.4% which varied between Aborting/non-pregnant (1.8%), age-matched pregnant controls (0.6%) and high fertility (2.1%) ewes. However, despite the variation observed, there was no statistical difference between the three groups. In addition, Neospora DNA was detected by PCR in 13% of submitted foetal brains and in ewe blood from aborting/non-pregnant (6.9%), age-matched pregnant controls (3.6%) and high fertility pregnant (2.1%) ewes. When the PCR results were considered with the IFAT and IDEXX ELISA results, there was no correlation between serology positive and PCR positive blood samples. Taken together, these results reveal that reliance on ELISA-based serology or PCR alone may underestimate the involvement of Neospora. Furthermore, determining the involvement of Neospora appears to require a multi-facetted approach where diagnostic methods and serological cut-off values may need to be adjusted as further information about the effect of natural infections with N. caninum in the ovine host is elucidated.     

Induced transplacental transmission of Neospora caninum in cattle.

Three Jersey cows were inoculated SC and IM with 26 million Neospora caninum tachyzoites at 129 (cow 1), 126 (cow 2), and 81 (cow 3) days after mating. Cows remained clinically normal for at least 1 month after inoculation of N caninum. Cow 1 was euthanatized 32 days after inoculation because of gangrenous mastitis. Cow 1 had a live fetus with no gross lesions; however, microscopic lesions were seen in the fetus and consisted of severe nonsuppurative necrotizing encephalitis of the cerebral white matter. Neospora caninum was identified in lesions by staining with anti-N caninum serum in an immunohistochemical test, by bioassays in mice, and by inoculation of bovine monocyte cultures with fetal tissue homogenate. Neither N caninum nor lesions were associated with infection with the protozoon identified in tissues of cow 1. Cows 2 and 3 aborted small autolysed fetuses 101 and 74 days, respectively, after inoculation with N caninum; the fetuses and attached placenta were unsuitable for laboratory investigations. Cows 2 and 3 remained clinically normal 4 months after abortion. Results of this study indicated that N caninum can be transmitted transplacentally in cattle.     

Occurrence of Neospora caninum and Toxoplasma gondii infections in ovine and caprine abortions

Neospora caninum and Toxoplasma gondii are closely related cyst-forming apicomplexan parasites identified as important causes of reproductive failure in cattle and small ruminants, respectively. Protozoan abortion in small ruminants is traditionally associated with T. gondii, but the importance of N. caninum remains uncertain. The aim of this study was to investigate the presence of N. caninum and T. gondii infections in abortion cases in small ruminants submitted for diagnosis. For this purpose, 74 ovine and 26 caprine aborted foetuses were recovered from different areas in Spain. Foetal histopathology was used to detect the presence of protozoal-associated lesions in brain. The presence of N. caninum and T. gondii was confirmed by PCR. Protozoal infection was detected in 17 out of 100 (17%) foetuses examined by at least one of the diagnostic techniques used. Lesions suggestive of protozoal infection were observed in 10.8% (8/74) and 15.4% (4/26) of the ovine and caprine abortions respectively. N. caninum and T. gondii infection was detected by PCR in 6.8% (5/74) and 5.4% (4/74) of sheep foetuses, respectively, of which five showed protozoal-associated lesions. N. caninum DNA was detected in 11.5% (3/26) of goat foetuses, of which two showed protozoal-associated lesions, whereas T. gondii DNA was detected in one goat foetus with no lesions. The simultaneous presence of N. caninum and T. gondii DNA was detected in one sheep foetus with severe lesions. This study demonstrates that N. caninum plays a significant role in abortion in small ruminants in the studied population. In addition, our results highlight the importance of differentiating between protozoa whenever characteristic lesions are observed.     

Chlamydia-related abortions in cattle from Graubunden, Switzerland

In 2001, the first case of bovine chlamydial abortion was reported in canton Graubunden, Switzerland. In this region, Chlamydophila (Cp.) abortus is endemic in small ruminants. Hence, we aimed to investigate the incidence of chlamydia-related abortions in cattle from Graubunden. During breeding seasons of 2003-2004, formalin-fixed and paraffin-embedded placenta specimens (n = 235) from late-term abortions in cattle were analyzed by histopathology, immunohistochemistry with a Chlamydiaceae-specific monoclonal antibody against chlamydial lipopolysaccharide (LPS), and 2 different polymerase chain reaction (PCR) methods (16 S ribosomal ribonucleic acid [rRNA] PCR, intergenic spacer [IGS-S] PCR), followed by PCR product sequencing. In 149 of 235 cases (63.4%), histopathologic lesions such as purulent and/or necrotizing placentitis were observed. Chlamydial antigen was clearly demonstrated in immunohistochemistry in only 1 of 235 cases (0.4%). Cp. abortus or Cp. psittaci was found in 12 of 235 (5.1%) and 10 of 235 cases (4.2%) by 16 S rRNA PCR and IGS-S PCR, respectively. However, we detected, by 16 S rRNA PCR, 43 of 235 cases (18.3%) to be positive for chlamydia-like organisms. In contrast to the situation in small ruminants in the canton Graubunden, bovine abortion from Cp. abortus seems not to play an important role. Nevertheless, zoonotic potential should be taken into account when handling abortion material from cattle. The significance of chlamydia-like isolates other than Waddlia chondrophila remains an open question in abortion and needs further investigation.     

Neospora caninum seroprevalence in dairy cattle in central Thailand

The seroprevalence, in dairy cattle, of antibodies to Neospora caninum, the relationship between seropositivity and age (heifer versus cow), the relationship of herd infection with herd size and the relationship of herd infection with the presence of dogs on the farm were studied. The study involved 549 cows and 82 dogs in 59 dairy herds in Nakhon Pathom, Thailand. A competitive enzyme-linked immunosorbent assay (cELISA) with NC-specific monoclonal antibody was used to detect the NC antibodies in the sera. Individual and herd seroprevalence of NC were 5.5% (30/549) and 34% (20/59), respectively. No significant relationships between NC seropositivity with the age of the cows (heifer versus cow; P > 0.05) and between herd infection and the presence of dogs on the farm (P > 0.05) were found. Herd size significantly affected herd infection (P < 0.05) with higher infection in large than small herds (> or = 21 versus < or = 20 cows). Of 12 cows with a history of abortion, one was seropositive to NC. The seroprevalence of NC antibodies in dogs was 1.2% (1/82). This is the first NC seroprevalence study in dogs in Thailand. It was concluded that Neospora infection was more common at the herd level rather than the individual level in Thailand and the presence of dogs on the farm was not related to the level of herd infection. Caution should be taken in the interpretation of serological tests from the farm dogs.     

Epidemiological investigations of abortions due to Neospora caninum on Swiss dairy farms

Abortions apparently due to Neospora caninum in two Swiss dairy herds were investigated by means of a PCR and ELISA, and other potential causes were eliminated. In addition, a case-control study of 24 case herds and 24 control herds indicated that N caninum-associated abortions were more likely to occur in herds with antibodies to Coxiella burnetii (with an odds ratio [OR] of 3.38 with a 95 per cent confidence interval [CI] of 1.82 to 6.22), whereas the likelihood was less in herds with antibodies to Leptospira species (OR 0.34,95 per cent CI 0.13 to 0.75), or in herds with antibodies to bovine viral diarrhoea (BVD) virus (OR 0.75,95 per cent CI 0.56 to 1.00), or Chlamydia psittaci (OR 0.18,95 per cent CI 0.09 to 0.35), or in herds in which BVD virus had been isolated from an aborted fetus (OR 0.11,95 per cent CI 0.02 to 0-58).     

奶牛新孢子虫病血清流行病学调查

本研究用酶联免疫吸附试验（ELISA）对采集自北京地区的94份奶牛血清（随机采集）和河北地区的55份奶牛血清（有流产史奶牛），进行Neospora caninum血清抗体检测。结果发现，北京地区随机采集的奶牛血清N．caninum抗体阳性率为18．1％（17／94），河北地区有流产史的奶牛N．caninum血清抗体阳性率为23．6％（13／55）。采用牛奶记录体系（DHI）对北京地区17头N．caninum血清抗体阳性牛进行了日产奶量、乳中蛋白率和乳脂率的测定，并与同群牛中134头阴性牛比较。结果表明，N．caninum血清抗体阳性牛日产奶量比阴性牛降低9．7％，乳中蛋白率和乳脂率分别降低20％和15．4％。初步证明N．caninum血清抗体阳性奶牛产奶量降低及奶品质的下降。对不同N．caninum抗体滴度阳性牛的泌乳期主要生产性能比较发现，其生产性能的变化与抗体滴度无明显相关性。     

Neospora caninum serostatus and culling of Holstein cattle

OBJECTIVE: To determine whether time until culling or risk of culling was associated with Neospora caninum serostatus among Holstein cattle in dairy herds in Ontario. DESIGN: Retrospective cohort study. ANIMALS: 3,412 cows in 56 herds. PROCEDURE: Blood samples were collected, and serum was tested for antibodies against N. caninum. Information on cows that were culled was collected during the 1- to 2-year period that producers were unaware of serostatus of individual cows in their herds. RESULTS: Herd prevalence of N. caninum-seropositive cows ranged from 0 to 68.3% (median, 7.0%). During the time of the study, 184 of 359 (51.3%) N. caninum-seropositive cows were culled, compared with 1,388 of 3,053 (45.5%) seronegative cows. Mean time from blood sample collection to culling for seronegative cows (289 days; 95% confidence interval, 280 to 299 days) was not significantly different from mean time for seropositive cows (296 days; 95% confidence interval, 269 to 323 days). Survival analysis indicated that N. caninum serostatus was not associated with time to culling or risk of culling. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that N. caninum serostatus of Holstein cows in Ontario was not significantly associated with either time to culling or risk of culling. Thus, N. caninum serostatus alone should not be used to determine whether cows should be culled.