Seroprevalence and associated risk factors of neosporosis in beef and dairy cattle in Italy

A cross-sectional serological survey for Neospora caninum was carried out on beef and dairy cattle in southern and northern Italy. A total of 111 herds and 1140 animals were tested using an ELISA assay (CHEKIT-Neospora) to detect anti-N. caninum antibodies. Management and individual data were collected and analysed both by linear and logistic multiple-regression models in order to find good predictors of the cattle seroprevalence and anti-N. caninum antibody level. At least one seropositive animal was found in 49 herds (44.1%), of which 31 (39.7%) from southern Italy and 18 (54.5%) from northern Italy. A total of 126 head of cattle (11%) were found to be seropositive and the seroprevalence was lower in southern (8.7%) than in northern Italy (16%). One of the best predictors of neosporosis seroprevalence in this study was the practice of self-rearing replacement heifers. Further risk factors were linked to higher stocking density, i.e. animals farmed in large herds and with no summer or permanent grazing practices were more likely to be seropositive than others. Farms with two or more dogs had higher herd seropositivity than farms with one or no dogs and this factor interacted significantly with the farm size and presence of poultry. Among individual characteristics, seropositivity was higher in animals sampled in mid- or late-pregnancy compared to animals either in early pregnancy or not pregnant. There was a significant interaction between the factors for pregnancy status and grazing practices. None of the epidemiological data recorded was a good predictor of the anti-N. caninum antibody level.     

Prevalence of serum antibodies to bovine herpesvirus-1 and bovine viral diarrhea virus in beef cattle in Uruguay

Our objective was to determine the prevalence of serum antibodies to bovine herpesvirus-1 (BHV-1) and bovine viral diarrhea (BVD) virus in beef cattle in Uruguay. A random sample of 230 herds selected with probability proportional to population size based on the number of cattle was chosen from a list frame of all registered livestock farms as of June 1999. Sera from up to 10 heifers, cows and bulls (up to 30 sera total per herd) were collected on selected farms between March 2000 and March 2001 and evaluated by means of enzyme-linked immunosorbent assays (ELISAs). Overall, 6358 serum samples were evaluated. We also collected data on previous diagnosis of BHV-1 or BVD infections and on the use of vaccines against these agents.

The estimated prevalence of exposure to BHV-1 and BVD at the herd level for the Uruguayan beef population was 99% and 100%, respectively. Approximately 37% of beef cattle in Uruguay have been exposed to BHV-1 and 69% to BVD virus. Only 3% of beef herds in Uruguay regularly (typically, annually) use vaccines against either of these agents.     

Prevalence of bovid herpesvirus-4 and its antibody in cattle in Minnesota

Serologic analyses and virus isolation studies were carried out to determine the role of bovid herpesvirus-4 (BHV-4) in infections in cattle, principally those of the reproductive tract. Serologic analyses were performed, using an indirect fluorescent antibody test on thoracic fluid specimens from aborted fetuses and on sera from 3 sources of adult cattle. Virus isolation was attempted from field cases of abortion, early embryo death, and postpartum vulvovaginitis/metritis, using uterine discharge and buffy coat preparations obtained from cows and tissues obtained from aborted fetuses. Of 420 fetal thoracic fluid specimens examined, 5 were positive for BHV-4 antibodies. Seventeen percent of adult cattle from 2 sources ie, clinically normal herds and abattoir cattle, were seropositive for BHV-4 antibodies. Cattle from a third source, 4 herds with high incidence of reproductive tract disorders, had a seroprevalence rate between 36 and 88%. Two isolates of BHV-4 were also obtained from this group. The overall incidence of BHV-4 antibodies in clinically normal cattle was higher than previously recognized, with relatively higher prevalence in herds having reproductive problems (chi 2 = 156.5, P less than 0.005). At least 10% of the BHV-4 antibody-positive sera did not have neutralizing antibody against bovine viral diarrhea virus and/or bovid herpesvirus-1, both important causes of bovine reproductive tract disorders.     

Virological and serological evidence of bovine herpesvirus type 4 in cattle in Northern Ireland

Bovine herpesvirus type 4 (BHV-4), a member of the genus Rhadinovirus, subfamily Gammaherpesvirinae, within the family Herpesviridae, was isolated in fetal bovine lung cells from samples of vaginal discharge taken from a dairy herd in which approximately 50 per cent of the cattle developed metritis after calving. The identity of the isolate was confirmed by immunofluorescent staining with a BHV-4-specific monoclonal antibody and partial sequencing of a portion of the glycoprotein B gene. Serological testing failed to demonstrate a significant association between the exposure of the cattle to BHV-4 and the metritis, but several cattle seroconverted during the periparturient period, consistent with the recrudescence and shedding of virus associated with the stresses of parturition and the onset of lactation. Despite the previous failure to detect BHV-4 in Northern Ireland, a serological survey of 999 cattle in 49 dairy herds and 51 beef herds found widespread evidence of exposure: 29 of the dairy herds and 35 of the beef herds contained one or more seropositive cattle, and 33.3 per cent of the dairy cattle and 23.3 per cent of the beef cattle were positive.     

Prevalence of bovine herpesvirus 1 (BHV-1) infection in Hungarian cattle herds.

Hungarian cattle herds were surveyed for bovine herpesvirus 1 (BHV-1) infection by ELISA of milk and serum samples. In 1993, 75% of the large cattle herds (consisting of more than 50 cattle) and all small herds (small-scale producers' stocks), while in 1997 90% of the small herds were included in the survey. In the case of large herds, 79.3% of the herds and 64.1% of the samples tested were found to be positive. Of the small herds, 13.5% and 15.7% tested positive in 1993 and 1997, respectively. The majority of large herds were Holstein-Friesian dairy stocks. Small herds with an infection rate markedly exceeding the average were found in those counties where the small herds had been in close contact with the large-scale farms, or where new herds were established by using animals of uncontrolled infectious bovine rhinotracheitis (IBR) status originating from large farms. Attention is called to the importance of maintaining the IBR-free status of small herds that constitute one-third of the Hungarian cattle population.     

An association exists between bovine herpesvirus-4 seropositivity and abortion in cows

The prevalence of cattle seropositive to bovine herpesvirus-4 (BHV-4) is high in Belgium. In Belgian farms, clinical signs associated with BHV-4 infection essentially involve the genital tract and consist mainly of postpartum metritis or metroperitonitis. The role of BHV-4 in abortion has not been definitively demonstrated but epidemiological and experimental facts suggest its involvement. A seroepidemiological investigation was therefore conducted as a case-control study to compare the seroprevalences of BHV-4 infections in the aborted-cow population and in a randomly selected control group in the province of Liège (Belgium). The seroprevalence (17.2%) in aborted cows was significantly higher than that of the control group (10.0%). The odds ratio (OR) was 1.87 (1.06 < 3.30). BHV-4 infection is therefore considered as a risk factor for abortion in cows.     

Serological evidence of Neospora caninum in dual-purpose cattle herds in Venezuela

Bovine neosporosis is a parasitic disease produced by Neospora caninum that induces abortion in cows, and consequently has a negative impact on the herd's reproductive efficiency. The main objective of this research was to determine the serological evidence of N. caninum in cattle herds from Venezuela using an indirect antibody capture ELISA test. Four hundred and fifty-nine (459) serum samples from crossbred adult cows were collected to be tested for Neospora antibodies. The sampled cows came from 15 large farms located in eight important cattle states that have predominant dual-purpose production systems (cattle from these farms are used for both milk and meat production). Fifty-two cows (11.3%) were seropositive to N. Caninum. Thirteen (86.7%) of 15 studied herds had cows seropositive to N. caninum. The average within-herd seroprevalence was 11.5% (range 3.8-36.7%). Cows that aborted in some of these farms had 2.71 (P: 0.009) greater odds to be seropositive when compared to cows that did not abort. Each one of the eight states represented in our study had seropositive animals. These results are the first evidence of exposure to N. caninum in Venezuelan cattle herds, indicating the possible circulation of this pathogen in the country. Further epidemiological studies should be granted to determine the spread of the disease in the Venezuelan cattle industry and its associated risk factors.     

Bulk milk testing for antibody seroprevalences to BVDV and BHV-1 in a rural region of Peru

Bulk milk from 60 herds of dairy cattle in a rural region in the central highlands of Peru was tested for antibodies to bovine viral-diarrhoea virus (BVDV) and bovine herpesvirus type 1 (BHV-1). None of the herds had been vaccinated against BVDV or BHV-1. Commercially available indirect ELISA-kits were used for antibody detection. True prevalences of BVDV and BHV-1 antibody-positive herds were 96 and 51%, respectively. A relatively low proportion of strongly positive herds suggests, however, a low prevalence of active BVDV infection. BVDV optical densities (ODs) in bulk milk increased with herd size--indicating a higher within-herd prevalence in the larger herds (probably, in part a consequence of a higher rate of animal movement into these herds). For BHV-1, this pattern was not found; a relatively high proportion of the herds was free from BHV-1 infection in each size category. This could indicate a low rate of reactivation of latent BHV-1 infection.     

BHV-1 eradication program in Bavaria: a marker-independent strategy

In Bavaria a BHV-1 eradication program was initiated in 1986 and was changed to a compulsory program in 1998. The eradication success increased progressively from < 50% in 1986 to 87% of the farms in 2002. BHV 1-free farms are controlled by bulk milk serology twice a year along with blood serology in animals that are negative but from herds where positive field virus infected animals are present. All serological tests are performed with an indirect ELISA test, all positive results are confirmed by a gB ELISA. Currently about 100.000 virus infected cattle are in Bavarian herds, approximately 80% of these animals are in heavily infected herds (> 10 infected animals). These herds comprise about 5% of all Bavarian herds. The eradication of the virus in these heavily infected herds is the most diifficult, whereas the prevention of new infections appears controllable. In this review current problems in BHV1 eradication are named and possible improvements are discussed.     

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 retrospective evaluation of a Bovine Herpesvirus-1 (BHV-1) antibody ELISA on bulk-tank milk samples for classification of the BHV-1 status of Danish dairy herds

Bulk-tank milk samples analysed in a Bovine Herpesvirus-1 (BHV-1) blocking ELISA are still in use in the Danish BHV-1 programme as a tool to classify dairy herds as BHV-1 infected or BHV-1 free herds. In this retrospective study, we used data from the Danish BHV-1 eradication campaign to evaluate performance characteristics of the BHV-1 blocking ELISA in 1039 BHV-1-seropositive and 502 repeatedly BHV-1-negative dairy herds using the results of blood testing of the individual animals as the true infection status. At a cut-off value of 30% blocking reaction, the herd-level relative sensitivity and relative specificity were 82 and 100%, respectively. The herd-level relative sensitivity depended on the within-herd prevalence of seropositive cows and the cut-off value in the assay, but not on the time interval (up to 90 days) between the collection of the bulk-tank milk sample and the individual serum samples. The BHV-1 blocking ELISA on bulk-tank milk could detect seropositive herds (few), with prevalence proportions as low as one seropositive cow out of 70 cows.     

First serological evidence of BHV-1 virus in Algerian dromedary camels: Seroprevalence and associated risk factors

Infectious bovine rhinotracheitis (IBR), caused by bovine herpesvirus-1 (BHV-1), is a major livestock health concern in many countries of the world. The objectives of this cross-sectional study were (i) to estimate the seroprevalence of BHV-1 infection and (ii) to assess risk factors associated with this disease in dromedary camels in four districts of Algeria. Blood samples were taken from 865 camels from 84 randomly selected herds, and serum was analyzed for presence of antibodies against BHV-1 by indirect enzyme linked immunosorbent assay (ELISA). Logistic regression was used to determine associations between seroprevalence and potential risk factors (collected using a questionnaire). Antibodies against BHV-1 were detected in 3.7 % (32/865) of samples. Eighteen of 84 camel herds had at least one BHV-1 seropositive camel, giving a herd seroprevalence of 21.4 %. Based on univariate analysis, the introduction of purchased animals and contact with others animal herds appeared as major risk factors. By using multivariate analysis, the only important risk factor was introduction of new animals. This study provided, for the first time, evidence of BHV-1 infection in dromedary camels in Algeria; it also provided estimates of seroprevalence of this disease and suggests that camels may serve as a reservoir of BHV-1 for spread to other species.     

Epidemiology of leptospirosis in dairy farm workers in the Manawatu. Part II. A case-control study of high and low risk farms

Subsequent to a cross-sectional serological survey of Manawatu dairy farm workers, a case-control study was carried out to investigate the correlation between titres to leptospiral serovars in workers and those in cattle in their herds. A total of 52 herds was investigated, 25 of which were high risk where milkers had titres of 1:96 or greater, and 27 were case-controls where milkers had no detectable agglutinin titres at a minimum serum dilution of 1:24. The serological prevalence of titres to hardjo in cattle on high risk farms (76.5%) was significantly higher (P<0.05) than on the case-control farms (60.0%). The geometric mean titres of seropositive cattle on high risk farms were also significantly higher (P<0.01) than in the cattle from the case-control farms, especially in the younger cohorts. These findings suggest that there was active endemic hardjo infection in the two- to three-year-old cattle on the high risk farms. Titres to pomona were demonstrated in only 5.2% of the cattle from both types of farm. Workers with titres to pomona tended to be from farms on which stock, especially calves, were bought-in and pigs were kept. Conventional measures for protecting milkers from contact with infected urine appeared to be ineffective and it is concluded that prevention of leptospirosis in dairy farm workers can only be achieved by elimination of infection in the herd by vaccination of cattle.     

Aspects of bovine herpesvirus-1 infection in dairy and beef herds in the Republic of Ireland

Background

Infection with bovine herpesvirus-1 (BHV-1) causes a wide range of disease manifestations, including respiratory disease and abortion, with world-wide distribution. The primary objective of the present study was to describe aspects of BHV-1 infection and control on Irish farms, including herd-level seroprevalence (based on pooled sera) and vaccine usage.

Methods

The characteristics of a diagnostic indirect BHV-1 antibody ELISA test when used on serum pools were evaluated using laboratory replicates for use in the seroprevalence study. The output from this indirect ELISA was expressed as a percentage positivity (PP) value. A proposed cut off (PCO) PP was applied in a cross-sectional study of a stratified random sample of 1,175 Irish dairy and beef cattle herds in 2009, using serum pools, to estimate herd seroprevalence. The study was observational, based primarily on the analysis of existing samples, and only aggregated results were reported. For these reasons, ethical approval was not required. Bulk milk samples from a subset of 111 dairy herds were analysed using the same ELISA. Information regarding vaccine usage was determined in a telephone survey.

Results

A PCO PP of 7.88% was determined to give 97.1% sensitivity and 100% specificity relative to the use of the ELISA on individual sera giving maximization of the prevalence independent Youden''s index, on receiver operating characteristics analysis of replicate results. The herd-level BHV-1 seroprevalence was 74.9% (95% CI - 69.9%-79.8%), with no significant difference between dairy and beef herds. 95.5% agreement in herd classification was found between bulk milk and serum pools. Only 1.8 percent of farmers used BHV-1 marker vaccine, 80% of which was live while 75% of vaccinated herds were dairy.A significant association was found between herd size (quartiles) and seroprevalence (quartiles).

Conclusions

The results from this study indicate BHV-1 infection is endemic, although BHV-1 vaccines are rarely used, in the cattle population in Ireland.     

BVDV and BHV-1 infections in dairy herds in northern and northeastern Thailand

Bulk milk samples from 220 dairy herds were collected at 9 public milk collection centres in the northeastern and northern Thailand, and a subset of 11 herds was selected for individual testing. The samples were tested for presence of antibodies to BVDV and BHV-1 using an indirect ELISA. The results from the bulk milk testing demonstrated a moderate level of exposure to BVDV and BHV-1 (73% and 67%, respectively). However, the low proportion of herds with high BVDV antibody-levels (13%) and the low within-herd seroprevalence of BVDV and BHV-1 in the 11 herds (24% and 5%, respectively), particularly among the young stock (15% and 0%, respectively), demonstrated a low prevalence of active BVDV infection and a low rate of reactivation of latent BHV-1. The presence of a self-clearance process was also indicated by the results from the individual testing. Moreover, a surprisingly low prevalence of BVDV and BHV-1 antibody-positive herds at one of the milk centres was found. This centre was established 5-10 years before the others. Our impression is that this reflects the self-clearance process, where consecutive replacement of imported infected animals without further spread has resulted in a nearly total elimination of the infections. Based on our experiences and on these results we are convinced that this process can continue if there is awareness of herd biosecurity. This is especially important in the context of a future intensification of the dairy production.     

Epidemiology of leptospirosis in dairy farm workers in the Manawatu. Part II. A case-control study of high and low risk farms

Subsequent to a cross-sectional serological survey of Manawatu dairy farm workers, a case-control study was carried out to investigate the correlation between titres to leptospiral serovars in workers and those in cattle in their herds. A total of 52 herds was investigated, 25 of which were ‘high risk’ where milkers had titres of 1:96 or greater, and 27 were case-controls where milkers had no detectable agglutinin titres at a minimum serum dilution of 1:24.

The serological prevalence of titres to hardjo in cattle on ‘high risk’ farms (76.5%) was significantly higher (P<0.05) than on the case-control farms (60.0%). The geometric mean titres of seropositive cattle on ‘high risk’ farms were also significantly higher (P<0.0l) than in the cattle from the case-control farms, especially in the younger cohorts. These findings suggest that there was active endemic hardjo infection in the two- to three-year-old cattle on the ‘high risk’ farms.

Titres to pomona were demonstrated in only 5.2% of the cattle from both types of farm. Workers with titres to pomona tended to be from farms on which stock, especially calves, were bought-in and pigs were kept.

Conventional measures for protecting milkers from contact with infected urine appeared to be ineffective and it is concluded that prevention of leptospirosis in dairy farm workers can only be achieved by elimination of infection in the herd by vaccination of cattle.     

Bovine herpesvirus type 1 abortions detected by a semi-nested PCR in Brazilian cattle herds

Bovine herpesvirus type 1 (BHV-1) was investigated by a semi-nested polymerase chain reaction (SN-PCR) and by MDBK cell culture virus isolation in organ fragments from 55 aborted fetuses collected from beef and dairy cattle herds with history of reproductive problems in the North of Paraná State, Brazil. A 425 bp amplicon of the BHV-1 glycoprotein D gene was detected in 14 (25.4%) aborted fetuses. BHV-1 was isolated in MDBK cells from the tissue of 5 (9.1%) fetuses. The specificity of positive results was evaluated by Restriction Fragment Length Polymorphism (RFLP) with Bgl I restriction of DNA amplified by SN-PCR, and by virus-neutralization and immunofluorescence with rabbit anti-BHV-1 polyclonal antibodies for virus isolated in cell culture. The results of this work demonstrate the importance of using other diagnostic techniques, like SN-PCR, for BHV-1 detection in organ fragments from aborted fetuses and the high frequency of this virus in reproductive failures in Brazilian cattle herds.     

Herd management practices associated with paratuberculosis seroprevalence in Dutch dairy herds

We describe the paratuberculosis management practices applied in dairy herds in the Netherlands. The findings from paratuberculosis seronegative and seropositive herds were compared to discover possible risk factors. In total, 370 randomly selected herds with > or =20 dairy cows were surveyed. A questionnaire was used to collect data on current and previous paratuberculosis management practices. All cattle aged > or =3 years were serologically tested for paratuberculosis using an enzyme-linked immunosorbent assay. Herds with >33 tested cattle, of which only one was seropositive, were excluded to reduce the risk of including false-positive herds in the analysis. A comparison of the management data of the seronegative herds (n = 166) and the seropositive herds (n = 143) showed that in both groups important management measures for the prevention of paratuberculosis, such as calving in a cleaned calving area, removing the calf immediately after birth, and feeding paratuberculosis non-suspect roughage to calves, were used only rarely. However, such measures should be regarded as the critical first step to control the disease and/or reduce its prevalence. Using univariable analysis, four factors were statistically different between seronegative and seropositive herds: herd size, cows with clinical signs of paratuberculosis, prompt selling of clinically diseased cattle and feeding milk replacer. Using a multivariable logistic regression model, only herd size was a significantly different factor. These results indicate that most of the paratuberculosis preventive management measures were executed on these Dutch dairy farms only to a limited extent.     

Virological and serological studies on the role of PI-3 virus, BRSV, BVDV and BHV-1 on respiratory infections of cattle. I. The detection of etiological agents by direct immunofluorescence technique

Nasal cells extracted from nasal swabs obtained from 95 cattle with signs of respiratory disease, out of eleven different herds, were tested for BHV-1, PI-3 virus, BRSV and BVDV using direct immunofluorescence technique. Viral antigen positive samples were detected in seven out of eleven herds examined. Of the 95 individual diseased cattle, 19 were found positive for at least one viral antigen. It was found that especially BHV-1 and PI-3 virus are important causative agents in cattle respiratory disease, both or in combination with other pathogenic agents. Multiple infection in virologically positive herds were observed in six (9.8%) of 61 animals tested. The findings reveal that single or multiple infections of selected viruses may be present in an important range in cattle and that direct immunofluorescence technique as a rapid method, based on the detection of viral antigen in nasal swab samples, is useful to establish the viral aetiology of acute bovine respiratory disease caused by these viruses, particularly in the diagnosis of mixed viral infections.     

Bovine herpesvirus type 1 (BHV-1) and bovine viral diarrhoea virus (BVDV) infections in dairy herds: self clearance and the detection of seroconversions against a new atypical pestivirus

The epidemiology of bovine herpesvirus type 1 (BHV-1) and bovine viral diarrhoea virus (BVDV) was studied in a population of small dairy herds that had not been vaccinated. Bulk tank milk samples of 186 herds in Thailand were collected four times between 2002 and 2004. Serum samples from individual animals in 11 herds were also taken on three occasions. The prevalence of BHV-1 in the 186 herds was 61% in 2002, decreasing to 48% in 2004 and for BVDV was 91% in 2002, decreasing to 72% in 2004. A BVDV antigen-positive calf was found in one of the 11 herds, and animals in this herd and three other herds seroconverted to a recently described atypical BVDV strain (HoBi). This study showed a significantly decreasing prevalence for both BHV-1 and BVDV due to a self-clearance process. Further studies are needed to find out how the atypical BVDV strain entered the cattle population.