A longitudinal study of Neospora caninum infection on a dairy farm in New Zealand

A 600-cow New Zealand dairy herd experienced an abortion storm in 1997 and was monitored (blood sampling at about 3-month intervals) from May 1997 until January 1999. Abortion risk reached 9% in 1997 and was highest in heifers at 19%. The abortion risk decreased in 1998 to 3.2% (still somewhat higher than during the years prior to the outbreak). The serological reaction pattern for Neospora caninum showed an association with abortion risk only around the time of the 1997 outbreak when seropositive cows were 4.2 times more likely to abort than negative ones. Over the whole study period, only 27% of cows that were sampled on all nine visits always tested negative. Offspring from dams which had positive tests for Neospora caninum were 2.4 times more likely to abort than those from dams testing consistently negative. Controlling for age and breed, seropositive cows produced more milk than those that were consistently negative. Infection might have been present endemically within this herd prior to the epidemic, but in 1997 an additional factor appeared to have triggered the outbreak.     

Herd-level risk factors for Neospora caninum seroprevalence in dairy farms in southern Brazil

A cross-sectional study was used to test the relationship between herd seroprevalence to Neospora caninum and various potential herd-level risk factors in 60 dairy farms located in two distinct regions in southern Brazil. Thirty farms were randomly selected from within each region. A questionnaire was designed to summarize each farm's production system as it might relate to N. caninum transmission. The questionnaire contained 105 closed questions relating to general characteristics of the farms, farm facilities, management, source of food and water, herd health, environment and biosecurity, which included questions relevant to N. caninum transmission, including presence and number of dogs and other animals, purchase of animals and contact with man. Serum samples were collected from 40% of animals in each farm and N. caninum antibodies were detected by immunofluorescent antibody test (IFAT). The association between potential risk factors and the probability of an animal being seropositive was modeled using a generalized estimation equations (GEE) logistic regression model. The model accounted for multilevel correlation of data from multiple animals within herds. The mean (+/-S.D.) number of animals in the 60 herds was 64.5 (+/-45.6), ranging from 20 to 280 females. Blood samples were collected from 1549 animals. The size of the farms varied from 4 to 100 ha (mean 30.1+/-25.9 ha). At least one dog was found in 57 of the 60 dairy farms (95%). The mean number of dogs was 3.1 (+/-1.9), ranging from 0 to 10. All females were raised on pasture. For all cattle sampled, N. caninum seroprevalence was 17.8%. Overall, 93.3% of herds (56/60) had at least one seropositive animal identified. Four variables were significantly associated with N. caninum sero-response in the 57 dairy farms, which were included in the final multivariable model: the number of dogs on the farm, farm area (hectares), feeding pooled sources of colostrum and region. The odds of a cow being seropositive increased 1.13 times for each additional dog present on the farm (P=0.021). Cattle from farms that fed calves colostrum pooled from multiple cows had 1.79 times greater odds for being seropositive for N. caninum (P<0.003). The probability of being seropositive was inverse to the area of the farms, such that cattle had 0.92 times the odds to be seropositive (P=0.014) for each additional 10 ha of farmland. Finally, cattle from farms in region one had 0.71 times the odds to be seropositive than cattle from region two (P=0.035). Results of this study suggest that several risk factors may explain why dairy cattle in Brazil may become exposed to N. caninum. However, further investigation of these factors is necessary because the purpose of this study was to refine and generate hypotheses on N. caninum transmission.     

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.     

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.     

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.     

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).     

Seroepidemiological study of Neospora caninum infection in dogs found in dairy farms and urban areas of Aguascalientes, Mexico

The objective of this study was to describe the seroepidemiology of Neospora caninum infection in dogs in close contact with dairy cattle and dogs from urban areas of Aguascalientes, Mexico, as well as to estimate the possible association between age, gender and size of these groups and seroprevalence. Sera were obtained from samples of 152 dogs in dairy farms and 116 in the urban area, and were subjected to ELISA test. General seroprevalence reached 32%, while the seroprevalence in farm dogs (41%) was significantly higher than in dogs from the urban area (20%) (p<0.05). Regarding age groups, general seroprevalence was greater among dogs between 11 and 15 years of age (67%) remaining equal between males and females (32 and 31%, respectively). Nevertheless, females resident in farms had a higher seroprevalence (42%) than female resident in the urban area. Regarding size, large-sized animals had greater seroprevalence in farms (58%), while in the urban area medium-sized animals were those that had the highest seroprevalence (27%). It was found that dairy farm dogs had a higher risk of infection (OR=2.79; p=0.0004), and that in said group, the age range from <1 to 5 years of age was identified as a risk factor (OR=3.11, p=0.001).     

Frequency of infection by Neospora caninum in wild rodents associated with dairy farms in Aguascalientes,Mexico

The purpose of this study was to detect the presence of Neospora caninum in tissues from wild rodents associated with dairy farms in Aguascalientes, Mexico. Individuals from Mus musculus, Rattus novergicus and Spermophilus variegatus were caught in dairy farms with history of positive status to N. caninum. A total of 33 rodents were caught from which samples were taken of brain, spinal cord, liver and heart and were tested with nested PCR and Immunohistochemistry (IHC). Results obtained with PCR showed that M. musculus had 77% (10/13) positive samples to N. caninum, S. variegatus had 71% (10/14) and R. novergicus had 50% (3/6). The results obtained with IHC showed that 15% (2/13) of M. musculus individuals were positive, while 43% (6/14) of S. variegatus and 33% (2/6) R. novergicus were also positive. This is the first report of the presence of N. caninum in S. variegatus.     

Seroprevalence of Neospora caninum in dairy cattle in Bahia, Brazil.

Sera collected from 447 dairy cattle on 14 dairy farms were tested for Neospora caninum antibodies by use of an immunofluorescent antibody technique. Positive reactions with titres > or =1:200 were found in 63 (14.09%) of animals. Neospora positive sera were also tested for Toxoplasma gondii antibodies by using a commercial latex agglutination test. Antibodies to T. gondii were detected in 3 (4.76%) of 63 N. caninum positive sera. These results indicate that N. caninum infection is widespread among dairy cattle in Bahia state.     

Survey of Neospora caninum infection by nested PCR in aborted fetuses from dairy farms in Aguascalientes, Mexico

The aim of this study was to determine by nested PCR probe the presence of Neospora caninum in fetal brain tissue from aborted dairy fetuses in Aguascalientes, a state in the central part of Mexico. Forty-four fetal brains from eight dairy farms were analyzed using single tube nested PCR probe with primers NF1, NS2, NR1 and SR1, with histopathology as the reference technique. From the 44 bovine fetuses studied, 35 (80%) were diagnosed as being infected by N. caninum using PCR probe and 20 (45%) were considered positive by histopathology, because they had lesions suggestive of bovine neosporosis. Fair agreement was observed (31%) between both probes. The age of aborting cows ranged from 1.6 to 2.9-year-old in 17 (49%) of the positive cases diagnosed by PCR. Gestational ages of aborted fetuses ranged from <3 to 9 months, with average of 5.6 months, and no statistical significant difference was observed when ages of infected and no infected fetuses were compared (P > 0.05). The frequency of N. caninum positive cases observed could be considered to be higher than other areas of Mexico and other countries.     

Factors associated with seroprevalence of Neospora caninum in dairy cattle in southeastern Brazil

Neosporosis is an infectious disease caused by Neospora caninum, an obligate intracellular cyst-forming protozoan considered a major cause of miscarriage in dairy cattle in many parts of the world. This cross-sectional study evaluated the relationship between reproductive abnormalities and seropositivity to N. caninum in 1,204 dairy cows from 40 farms located in the state of Minas Gerais, southeastern Brazil. Producers were interviewed, and blood samples were collected to perform indirect immunofluorescence tests (IFAT 1:200). Association between reproductive abnormalities and seropositivity in cattle was evaluated with generalized estimating equations. The true herd-level seroprevalence of N. caninum was 95 % (83.3–99.1), while the individual-level true seroprevalence was 21.6 % (19.2–24.2). Several reproductive abnormalities were significantly associated with seropositivity to N. caninum: occurrence of repeated estrus (p?=?0.02; OR?=?3.84; 95 % CI?=?1.239–11.893), repeated miscarriages (p?=?0.001; OR?=?2.54; 95 % CI?=?1.423–5.402), and temporary anestrus (p?=?0.001; OR?=?3.44; 95 % CI?=?1.976–5.994). Furthermore, loose dogs (p?=?0.041; OR?=?2.20; 95 % CI?=?1.033–4.672) when fed raw meat (p?=?0.001; OR?=?1.91; 95 % CI?=?1.443–2.519) are risk factors for N. caninum infection. We observed that seropositivity to N. caninum in cattle increases risk of miscarriage by almost twice throughout the reproductive life of cows (p?=?0.004; OR?=?1.978; 95 % CI?=?1.249–3.131). Serologic evidence in this study indicates that N. caninum infection is widely distributed among dairy herds and significantly associated with reproductive disorders, especially miscarriage, repeated estrus, and temporary anestrus.     

Factors associated with infection by Neospora caninum in dogs in Brazil

From August 2006 to 2008, 411 dogs in northeastern Brazil were evaluated for seropositivity to Neospora caninum. The dogs were clinically examined, and their owners were interviewed about the conditions in which the animals were maintained in order to assess the factors associated with infection by this parasite. A serum sample was taken from each dog for serological examination in an indirect fluorescent antibody test for N. caninum. The Yates' Chi-square test or Fisher's exact test was used to select the variables for the multivariate logistic regression model. Seropositivity was detected in 9.26% of dogs. The seropositivity rates of dogs from different environments were 2.6% (4/156) in urban areas, 13.1% (28/214) in peri-urban areas, and 14.6% (6/41) in rural areas. Factors associated with seropositivity for N. caninum were the following: contact with other dogs, access to food outside the home and residing in the peri-urban or rural environments (p<0.05). Results of this study confirm that dogs in urban, rural and peri-urban areas of northeastern Brazil are exposed to N. caninum. Control measures to prevent infection of dogs in the studied region should be focused primarily on preventing access to potential sources of infection, which include environments with other dogs, bovines, and other small intermediate hosts, such as birds and rodents.     

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.     

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.     

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.