A longitudinal study of Neospora caninum infection on three dairy farms in Brazil

Neospora caninum is an apicomplexan protozoan parasite that is one of the most important infectious causes of abortion in both dairy and beef cattle in many countries. The objectives of this longitudinal study were to determine the prevalence, rates of vertical and horizontal transmission of N. caninum and hazard for culling of N. caninum-seropositive animals in three Brazilian dairy herds. Blood samples from all animals were collected nine times at each of the three farms over a two-year period. Serum was tested for antibodies against N. caninum using the indirect fluorescent antibody test with a cutoff value of 1:100. The percentage of N. caninum-positive samples at each sampling time ranged at Farm I from 3.32% to 11.71%, at Farm II from 3.90% to 22.06% and at Farm III from 3.90% to 22.06%. The number of positive serological reactions varied in relation to the number of repeated samples taken from individual animals at each farm. In all herds, there was a high degree (P<0.05) of association between the N. caninum serological status of dams and daughters. The seropositive conversion rate was estimated as 0.37%, 3.00% and 6.94% per 100 cow-years at Farms I, II and III, respectively. The seronegative conversion rate was estimated as 31.58% and 11.11% per 100 cow-years at Farms I and III, respectively. In all herds, there was no difference (P>0.05) in the culling rate between the cattle that were seropositive cattle and seronegative for N. caninum infection. The results from this study confirm the importance of vertical transmission in the epidemiology of N. caninum. Although a few positive seroconversions indicated horizontal transmission, it does not appear to be the major route of infection for N. caninum.     

A longitudinal study of seroprevalence and seroconversion of Neospora caninum infection in dairy cattle in northeast Thailand

A long-term study was carried out in 11 dairy herds in the Khon Kaen province of northeast Thailand between August 2001 and November 2004. The objective was to investigate seroprevalence dynamics of Neospora caninum infection in the herds and to demonstrate patterns of seroconversion in individual cattle. Each herd was visited once a year, in total four times, and sera from cattle > 3 months of age and farm dogs as well as a sample from the bulk milk were collected. All samples were analysed for presence of specific antibodies by an N. caninum iscom ELISA. The overall percentage of antibody-positive cattle was constant and varied only between 10 and 13% over the 4 years, but the variation in within-herd seroprevalence between herds was substantial. Two herds had > or = 20% seropositive animals at all samplings and consistently high bulk milk OD, whereas two herds had no seropositive animal at the last two samplings and low bulk milk OD. Five herds had a decreasing trend of within-herd seroprevalence, whereas the remaining six herds had a higher portion of test-positive individuals at the end of the study. A total of 424 individuals were sampled more than once; 344 (81%) and 32 (8%) were consistently antibody-negative and antibody-positive, respectively. The proportions of animals that changed from being seronegative to seropositive and from being seropositive to seronegative between the years were 3.9-4.6% and 19-39%, respectively. Apparent vertical and horizontal transmission rates were 58% (95% CI; 44-71%) and 5% (95% CI; 3-7%), respectively. In conclusion, the overall percentage of N. caninum antibody-positive cattle was constant over the years, but the within-herd seroprevalence varied substantially between the herds. Seroconversions were likely to occur in individual cattle although most animals had consistent serological status throughout the study.     

Serology of a Neospora abortion outbreak on a dairy farm in New Zealand: a case study

AIM: To 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. METHODS: Thirty-five cows, that had either aborted, carried mummified calves, were not pregnant or calved normally were: bled several times at regular intervals and the sera tested for Neospora antibodies in the indirect fluorescence antibody test (IFAT). RESULTS: Maximal IFAT titres of up to 1:4000 occurred within 6 weeks of the abortion outbreak, decreased over the next 2 months to < or = 1:200 and remained at this level until the next scheduled bleed a further 2 months later. A rise in titres was subsequently observed in the cows that had aborted or were not pregnant (at the time of the abortions) or had carried mummified foetuses. Seroconversion was also observed in some of the control cows, which had, up until then, remained seronegative. A dog and cat in contact with the cows in the herd investigated were, however, negative in the IFAT. CONCLUSIONS: Maximal serological titres in Neospora abortions are observed within weeks of the abortion event and then quickly return to very low levels. Subsequently, a recrudescence of titres can be observed in infected cows during the next pregnancy, without it being associated with repeat abortions.     

Neospora caninum infections in Australia and New Zealand

OBJECTIVE: To review the current state of knowledge of Neospora caninum infections with particular reference to Australia and New Zealand. PROCEDURE: Several databases were searched electronically including Medline, Current Contents, Vet CD using several key words (Neospora caninum, neosporosis, abortion, cattle, dogs) and authors names. References in original articles were also traced and use made of the author's own original research in the field. CONCLUSION: N caninum is recognised worldwide and is also widespread, in particular in dairy cattle, in Australia and New Zealand. It has been reported in both countries retrospectively (in dogs) from the early 1970s. Abortion storms in dairy herds appear to be the most common feature. Recent reports indicate that the dog is the definitive host, in which the sexual cycle is completed. Further studies are however required to establish important aspects of the epidemiology, such as mode of transmission. In the absence of an effective vaccine, the most effective control strategy is selective culling of infected animals and the prevention of access of dogs to expelled placentas and foetuses and, possibly, to raw beef.     

Chronic Neospora caninum infection and repeat abortion in dairy cows: a 3-year study

The serological status of Neospora was monitored in animals older than 6 months in a dairy herd with a 3-year history of prevalent N. caninum and N. caninum-associated abortions. The numbers of animals in the herd tested each year of the study period were 259, 222 and 231, respectively. A separate analysis was performed on the 122 animals persisting in the herd for the 3 years. The overall seroprevalence of N. caninum in the herd decreased from 31.7% in the first year to 24.8% in the second year and to 19.9% in the third year of the study, while the overall abortion rate decreased from 20.6% in the first year to 5.5% in the second year, and 9.9% in the third. These decreases occurred in response to control measures adopted from the second year onwards, such as culling Neospora-seropositive aborted animals and inseminating Neospora-seropositive dams with beef bull semen. Of the total number of abortions recorded in seropositive animals, 51% were repeat abortions that occurred in 36.8% of the animals with a previous history of abortion. The initial seroprevalence of Neospora in the 122 animals followed for the 3 years was 18%, increasing to 21.3% in the second and third years. Seroconversion only occurred in four animals during the second and third years of the study and abortion occurred only in seropositive individuals. Of the total number of abortions recorded in the 122 animals, 61.5% were repeat abortions that occurred in 26.7% of the animals with a previous history of abortion. These results indicate that Neospora seropositivity can be very stable through time and N. caninum infected cows can show a high rate of repeat abortions. The present data reinforce the idea that annual serological screening for Neospora can be an effective and rapid method of detecting N. caninum infection, such that control measures can be established at the farm level.     

Abortion epidemic in a dairy herd associated with horizontally transmitted Neospora caninum infection

A dairy herd experienced an abortion epidemic during which 43 per cent of the cows at risk aborted. Neospora caninum infection was demonstrated in four of six fetuses suitable for examination and the group of at-risk cows that aborted had significantly higher N. caninum antibody concentrations than the at-risk cows that delivered a live calf at term (P<0.001). The antibody concentrations in the cow herd were significantly higher than in the youngstock (P<0.001), and the concentrations in the youngstock increased significantly (P<0.001) with age. When seven months to a year old, the calves born at term to the at-risk cows had significantly higher (P=0.007) antibody concentrations than age-matched calves born before the epidemic. At the time of the epidemic, there was a significant increase in the antibody levels of the herd that was not consistent with vertical infection alone, indicating that there appeared to have been a sudden large increase in the incidence of horizontal postnatal transmission of N. caninum to the cow herd, or to the surviving offspring of the at-risk cows, or to both of these groups.     

Epidemiology and impact of Neospora caninum infection in three Queensland tropical dairy herds

This study investigated the epidemiology of Neospora caninum in three tropical dairy herds in North Queensland, Australia. All animals in the herds were bled, and the sera were tested by ELISA for N. caninum antibodies. Herd records were examined, and the number of calves carried to term and the number of abortions which occurred over the lifetime of each animal were recorded to determine the abortion rate for each animal. Pedigrees were constructed for two of the herds to investigate whether vertical transmission was occurring. The seroprevalence of N. caninum ranged from 23% to 34%. The abortion rate in seropositive animals was significantly (p < 0.001) higher than in seronegative animals in all three herds (12-20.1% cf. 3.6-7%). Overall, the probability of a calf being seropositive was 3.5 times higher when the dam was also seropositive than when the dam was seronegative. Subsequent selective breeding employed by one herd reduced the N. caninum seroprevalence from 23% to 5% over a 9-year period. This study shows that N. caninum infection is prevalent in North Queensland dairy cattle, and both post-natal infection and vertical transmission are common.     

Neospora caninum infection in three dogs

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

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.     

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.     

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。     

Quantification of vertical and horizontal transmission of Neospora caninum infection in Dutch dairy herds

Ninety-six of 108 randomly selected Dutch dairy herds had one or more cows with a positive serostatus for N. caninum. In these 96 herds, we have quantified the probabilities of vertical transmission (VT) and horizontal transmission (HT) of N. caninum infection by combining serostatus and pedigree data in 4091 dam-daughter pairs. The probability of animals infected by vertical transmission during pregnancy (Prob(VT)) was calculated as the proportion of seropositive daughters among daughters of seropositive dams. The probability of animals infected by horizontal transmission (Prob(HT)) was the proportion of seropositive daughters among daughters of seronegative dams. These probabilities were calculated after the frequencies of observed dam-daughter combinations were corrected for (1) imperfect test-characteristics, (2) underestimation of horizontal transmission in situations that seronegative dams were horizontally infected after the birth of their daughters and (3) overestimation of vertical transmission in situations that seronegative daughters born from seropositive dams were horizontally infected. The incidence rate for horizontal transmission was calculated based on Prob(HT) and the average age of the animals in these herds. Based on the analysis of dam-daughter serology, Prob(VT) was 61.8% (95% CI: 57.5-66.0%) and Prob(HT) was 3.3% (95% CI: 2.7-3.9%). After adjusting the observed frequencies for imperfect test-characteristics, underestimation of horizontal transmission and overestimation of vertical transmission, Prob(VT) decreased to 44.9% (95% CI: 40.0-49.9%) while Prob(HT) increased to 4.5% (95% CI: 3.9-5.2%). Prob(HT) corresponded with an incidence rate for horizontal transmission of 1.4 (95% CI: 1.2-1.7) infections per 100 cow-years at risk. When stratifying herds for the presence of farm dogs, Prob(HT) was higher (5.5% (95% CI: 4.6-6.4%)) in herds with farm dogs than in herds without farm dogs (2.3% (95% CI: 1.5-3.4%)). When stratifying for within-herd seroprevalence, Prob(HT) was higher (10.3% (95% CI: 8.6-12.2%)) in herds with high (> or =10%) within-herd seroprevalence compared with herds with low (<10%) within-herd seroprevalence (2.0% (95% CI: 1.5-2.6%)). Although there was this relation between Prob(HT) and within-herd seroprevalence (crude OR(PREV) = 5.7 (95% CI: 4.0-7.9)), in herds without farm dogs, this relationship was no longer statistical significant (OR(PREV|DOG-) = 1.9 (95% CI: 0.7-5.5)). It indicated that the association between seroprevalence and Prob(HT) depended largely on the presence of farm dogs. In addition, when looking for the presence of specific age-groups with significantly higher seroprevalence compared with the rest of the herd, there were 7 herds in which two or more horizontally-infected animals were present in specific age-groups. This was an indication of a recent point-source exposure to N. caninum. These results reiterate the current control strategies to apply strict dog-management measures as well as to minimize within-herd seroprevalence by monitoring serostatus of animals.     

Protozoal encephalomyelitis of dogs involving Neospora caninum and Toxoplasma gondii in New Zealand

In 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 from those of T. gondii using light microscopy, but electron microscopy confirmed their characteristic features. Although Neospora abortion in cattle has only recently been recognised in New Zealand, this study has shown that neosporosis has been present in dogs since at least 1972.     

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.     

Disseminated infection with Neospora caninum in a ten-year-old dog

Naturally-occurring neosporosis with multiple organ involvement was identified in a 10-year-old neutered male Basset hound dog. Clinical signs were first noticed 3 weeks prior to referral and consisted of crouched stance and mild pelvic limb ataxia. Dexamethasone administration had provided transient improvement. On presentation to the teaching hospital, clinical signs included depression, pelvic limb ataxia, inability to stand without assistance, and pain on deep palpation of the cervical and lumbar vertebral column. Lesions were found in the myocardium, liver, spleen, adrenal glands, brain, and spinal cord. Tachyzoites of Neospora caninum were found in the myocardium and adrenal glands. Organisms stained with anti-Neospora caninum, but not to anti-Toxoplasma gondii serum in an immunohistochemical test.     

Udder health in dairy cattle infected with Neospora caninum

Blood samples were collected from 3449 cows on 57 representative Ontario dairy herds during the summer of 1998 and analysed for antibody to Neospora caninum using an ELISA. Forty-eight herds (2742 cattle) contained at least one N. caninum-seropositive animal. Two composite milk samples were collected from all cattle: the first on the day of blood collection and the second 68 to 365 days later. All milk samples were submitted for bacteriological culture. Ontario Dairy Herd Improvement Corporation (DHI) data were available for 3162 cattle in the 57 herds at the time of bleeding. Furthermore, complete DHI data were available for 1658 cattle that were culled between 12 and 24 months following blood collection. Using a standardised ELISA sample-to-positive (S/P) cut-off of ≥0.45, the corrected seroprevalence was 8.2% overall and 10.1% within seropositive herds. At blood collection the odds of N. caninum-seropositive cows having a high linear score (≥4.0; equivalent to a somatic cell count ≥200,000 cells/ml) was 27% less than for seronegative animals. Similarly, at the time of culling, the odds of having a high linear score was 22% less in N. caninum-seropositive cattle. Overall, linear score was lower in N. caninum-seropositive cattle at culling. After controlling for herd, parity, days in milk, and the interval between collection of milk samples, the odds of N. caninum-seropositive cattle testing positive for an environmental pathogen (i.e. environmental Streptococcus species and coliforms) on the second milk sample was 56% less than for seronegative animals. The odds were 83% less at a higher ELISA S/P cut-off of ≥0.70. Finally, the odds of N. caninum-seropositive cattle developing a new infection with a major pathogen (environmental or contagious) were 60% less than seronegative cows using the higher ELISA S/P cut-off.     

Monensin use against Neospora caninum challenge in dairy cattle

Using a randomized controlled trial design, a randomly allocated intervention group of 15 cows received a slow-release bolus that delivered 100 days of monensin. The negative control group of 15 cows received a placebo bolus that was identical to the monensin bolus, except without the monensin. Two weeks after bolus administration, all cows were challenged with a 2 ml subcutaneous injection of a live tachyzoite suspension. Whole blood and serum samples were collected from each cow every week for the first month post-challenge, and then every 2 weeks for the next 2 months. The extracted DNA from whole blood was tested for the Nc-5 gene fragment of Neospora caninum using a quantitative real time polymerase chain reaction. Serum was tested for antibodies to N. caninum using the IDEXX ELISA. Cows treated with monensin boluses had a significantly lower humoral immune response than cows treated with placebo boluses at one time point post-challenge (week 4 post-challenge). However, when adjusting for repeated measures within cows, the P value for this humoral difference was 0.098. No DNA for N. caninum was detected in either group, likely due to study design features.     

Use of bulk milk for detection of Neospora caninum infection in dairy herds in Thailand

The relationship between the level of Neospora caninum antibodies in bulk milk and the seroprevalence in lactating cows was investigated. Bulk milk was also used to estimate the prevalence of N. caninum infection in dairy herds in the northeast and north Thailand. Bulk milk and individual serum from all lactating cows in 11 herds as well as 220 bulk milk samples from nine milk collection centres were analysed for presence of N. caninum antibodies using an iscom ELISA. In the 11 herds the bulk milk absorbances ranged between 0.04 and 0.89 and the seroprevalences varied between 0 and 46%. Five herds had milk absorbances below 0.20, among those were the two herds housing only seronegative lactating cows. In the remaining three herds with such low bulk milk absorbances one or two cows (5-14%) were seropositive. Six of the investigated herds had bulk milk absorbances above 0.20. In the two herds with the highest bulk milk absorbances more than 30% of the cows were seropositive. Using an absorbance of 0.20 to discriminate between negative and positive herds, 102 (46%) of 220 bulk milk samples were judged positive. There was no significant difference in mean bulk milk absorbance between the milk collection centres within each region. However, the proportion of herds with bulk milk absorbances > or =0.50 in the north was statistically (P < 0.01) higher than that in the northeast. It was concluded that bulk milk antibody testing can be used to identify N. caninum-infected herds and that N. caninum is a common infection in dairy herds in Thailand.     

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.