Seronegative Herds Research Articles

BRSV seroprevalence and associated risk factors on Dutch dairy farms

Bovine respiratory syncytial virus (BRSV) causes infections of the respiratory tract of cattle and is endemic in the Netherlands. We aimed to update our knowledge on the seroprevalence of BRSV in youngstock on Dutch dairy farms by performing a cross sectional study during the winter of 2021–2022 and a telephone survey with the farmers to map the most important risk factors for the introduction, presence, and circulation of BRSV. Of 671 sampled calves among 135 herds, we found a seropositivity of 75 % at calf level and 77 % on herd level. Risk factors appeared similar to those identified by others including a higher ratio of youngstock versus adult cows and suboptimal colostrum administration. We concluded that the BRSV seroprevalence at animal and herd level is still substantially high, confirming the endemic circulation of this virus on the majority of the Dutch dairy farms. But given that there is also a smaller number of seronegative herds and the available knowledge about biological sound risk factors for BRSV circulation, it could be worth investigating the feasibility of decreasing the seroprevalence of BRSV at herd level in the Netherlands taking into account the lessons learned from the Norwegian control program.

Neospora caninum in beef herds in New South Wales, Australia. 2: analysis of risk factors.

To determine the influence of farm-level and animal-level factors on the seroprevalence of antibodies to Neospora caninum and associations between seropositivity and reproductive outcomes. A questionnaire for a cross-sectional survey was posted to the 63 properties with a herd size ≥50 beef breeding cows that had participated in a previous seroprevalence study. Correspondence analysis, which does not appear to have been used previously in any Australian studies of livestock diseases, was used in conjunction with logistic regression to analyse the data. Geographic factors that increased the risk of seropositivity included higher rainfall North Coast location. Herd management factors that increased the risk of seropositivity included the use of Bos indicus genetics, cross-breeding and running several breeds in the one herd. Using fox control measures was found to be protective against infection with N. caninum. The risk of abortion was 12-fold greater in individual animals that were seropositive for N. caninum. Within a herd, the calving rate was 10.4% lower in herds with one or more N. caninum-positive animals (P = 0.03), but the difference in abortion rate was not significant between seropositive and seronegative herds (0.3% higher, P > 0.3). This study confirmed previous observations of increased risks for N. caninum seropositivity with being located in the coastal subtropics, some styles of herd management and canid exposure. In addition, it suggested that cross-breeding and proximity to an urban area may increase the risk, and that having pet dogs may reduce the risk of seropositivity.

Evaluation of the Johne’s disease risk assessment and management plan on dairy farms in Ontario, Canada

Johne’s disease (JD) is a production-limiting gastrointestinal disease in cattle. To minimize the effects of JD, the Ontario dairy industry launched the Ontario Johne’s Education and Management Assistance Program in 2010. As part of the program, trained veterinarians conducted a risk assessment and management plan (RAMP), an on-farm questionnaire where high RAMP scores are associated with high risk of JD transmission. Subsequently, veterinarians recommended farm-specific management practices for JD prevention. Milk or serum ELISA results from the milking herd were used to determine the herd ELISA status (HES) and within-herd prevalence. After 3.5 yr of implementation of the program, the aim of this study was to evaluate the associations among RAMP scores, HES, and recommendations. Data from 2,103 herds were available for the analyses. A zero-inflated negative binomial model for the prediction of the number of ELISA-positive animals per farm was built. The model included individual RAMP questions about purchasing animals in the logistic portion, indicating risks for between-herd transmission, and purchasing bulls, birth of calves outside the designated calving area, colostrum and milk feeding management, and adult cow environmental hygiene in the negative binomial portion, indicating risk factors for within-herd transmission. However, farms which fed low-risk milk compared with milk replacer had fewer seropositive animals. The model additionally included the JD herd history in the negative binomial and the logistic portion, indicating that herds with a JD herd history were more likely to have at least 1 positive animal and to have a higher number of positive animals. Generally, a positive association was noted between RAMP scores and the odds of receiving a recommendation for the respective risk area; however, the relationship was not always linear. For general JD risk and calving area risk, seropositive herds had higher odds of receiving recommendations compared with seronegative herds if the section scores were low. This study suggests that the RAMP is a valuable tool to assess the risk for JD transmission within and between herds and to determine farm-specific recommendations for JD prevention.

A case-control study of risk factors for bovine brucellosis seropositivity in Peninsular Malaysia.

Bovine brucellosis was first reported in Peninsular Malaysia in 1950. A subsequent survey conducted in the country revealed that the disease was widespread. Current knowledge on the potential risk factors for brucellosis occurrence on cattle farms in Malaysia is lacking. Therefore, we conducted a case-control study to identify the potential herd-level risk factors for bovine brucellosis occurrence in four states in the country, namely Kelantan, Pahang, Selangor and Negeri Sembilan. Thirty-five cases and 36 controls of herds were selected where data on farm management, biosecurity, medical history and public health were collected. Multivariable logistic regression identified that Brucella seropositive herds were more likely to; have some interaction with wildlife (OR 8.9, 95% CI = 1.59–50.05); originated from farms where multiple species such as buffalo/others (OR 41.8, 95% CI = 3.94–443.19) and goat/sheep (OR 8.9, 95%Cl = 1.10–71.83) were reared, practice extensive production system (OR 13.6, 95% CI 1.31–140.24) and have had episodes of abortion in the past (OR 51.8, 95% CI = 4.54–590.90) when compared to seronegative herds. Considering the lack of information on the epidemiology of bovine brucellosis in peninsular Malaysia and absence of information on preventing the inception or spread of the disease, this report could contribute to the on-going area-wise national brucellosis eradication program.

Increasing prevalence of Coxiella burnetii seropositive Danish dairy cattle herds.

A study based on bulk tank milk samples from 120 randomly selected dairy cattle herds was conducted to estimate the prevalence of Coxiella burnetii seropositive dairy herds, to describe the geographical distribution, and to identify risk factors. Using the CHEKIT Q-fever Antibody ELISA Test Kit (IDEXX), the study revealed a prevalence of 79.2% seropositive herds, 18.3% seronegative herds, and 2.5% serointermediate herds based on the instructions provided by the manufacturer. Multifactorial logistic regression showed statistically significant associations (P < 0.01) between C. burnetii seropositivity and increasing herd size (OR = 1.02 per cow increment) and increasing regional average number of cattle per dairy herd (OR = 1.02 per animal increment). Herds >150 cows had 17.9 times higher odds of testing positive compared to herds <80 cows. The regional average number of cattle herds per square kilometer was borderline significantly related to the occurrence of seropositive dairy herds (P = 0.06). The results indicate an increased prevalence of seropositive dairy herds since the previous survey in 2008 and an adverse impact of increasing herd size and cattle density on the risk of seropositivity.

Lungworm Infections in German Dairy Cattle Herds — Seroprevalence and GIS-Supported Risk Factor Analysis

In November 2008, a total of 19,910 bulk tank milk (BTM) samples were obtained from dairy farms from all over Germany, corresponding to about 20% of all German dairy herds, and analysed for antibodies against the bovine lungworm Dictyocaulus viviparus by use of the recombinant MSP-ELISA. A total number of 3,397 (17.1%; n = 19,910) BTM samples tested seropositive. The prevalences in individual German federal states varied between 0.0% and 31.2% positive herds. A geospatial map was drawn to show the distribution of seropositive and seronegative herds per postal code area. ELISA results were further analysed for associations with land-use and climate data. Bivariate statistical analysis was used to identify potential spatial risk factors for dictyocaulosis. Statistically significant positive associations were found between lungworm seropositive herds and the proportion of water bodies and grassed area per postal code area. Variables that showed a statistically significant association with a positive BTM test were included in a logistic regression model, which was further refined by controlled stepwise selection of variables. The low Pseudo R2 values (0.08 for the full model and 0.06 for the final model) and further evaluation of the model by ROC analysis indicate that additional, unrecorded factors (e.g. management factors) or random effects may substantially contribute to lungworm infections in dairy cows. Veterinarians should include lungworms in the differential diagnosis of respiratory disease in dairy cattle, particularly those at pasture. Monitoring of herds through BTM screening for antibodies can help farmers and veterinarians plan and implement appropriate control measures.

Evaluation ofCoxiella burnetiiStatus in Dairy Cattle Herds with Bulk-tank Milk Positive by ELISA and PCR

Bulk-tank milk (BTM) samples are frequently used to evaluate the health status of dairy livestock. A large-scale investigation carried out in BTM samples from dairy cattle herds from a Q fever-endemic region in Northern Spain revealed a high degree of exposure to Coxiella burnetii. This study was aimed at assessing the value of BTM samples analysis as an indicator of the C.burnetii status in dairy cattle herds. Three herds with BTM samples positive for C.burnetii by ELISA and PCR were selected, and blood, faeces and individual milk and BTM samples were analysed by serology and PCR. In spite of the high antibodies titres found in BTM samples, only one of the three farms presented an active infection by C.burnetii, as revealed by the presence of bacterial DNA in vaginal mucus and in environmental samples collected in the calving area, a seroprevalence around 40% in heifers and the seroconversion rate observed in cows. Results obtained indicated that the analysis of BTM samples is a good epidemiological tool at the population level that can be used to discriminate between seropositive and seronegative herds, but at the herd level, additional tests are necessary to evaluate whether Q fever is a potential problem in the farm. When Q fever is suspected in a cattle herd, sera from a small group of 1- to 3-year-old animals need to be analysed to investigate recent contact with C.burnetii.

A novel double recognition enzyme-linked immunosorbent assay based on the nucleocapsid protein for early detection of European porcine reproductive and respiratory syndrome virus infection

Precise and rapid detection of porcine reproductive respiratory syndrome virus (PRRSV) infection in swine farms is critical. Improvement of control procedures, such as testing incoming gilt and surveillance of seronegative herds requires more rapid and sensitive methods. However, standard serological techniques detect mainly IgG antibodies. A double recognition enzyme-linked immunosorbent assay (DR-ELISA) was developed for detection of antibodies specific to European PRRSV. This new assay can recognize both IgM and IgG antibodies to PRSSV which might be useful for detecting in routine surveillance assays pigs that are in the very early stages of infection and missed by conventional assays detecting only IgG antibodies. DR-ELISA is based on the double recognition of antigen by antibody. In this study, the recombinant nucleocapsid protein (N) of PRRSV was used both as the coating and the enzyme-conjugated antigen. To evaluate the sensitivity of the assay at early stages of the infection, sera from 69 pigs infected with PRRSV were collected during successive days post infection (pi) and tested. While standard methods showed low sensitivity rates before day 14 pi, DR-ELISA detected 88.4% seropositive samples at day 7 showing greater sensitivity at early stages of the infection. Further studies were carried out to assess the efficiency of the new assay, and the results showed DR-ELISA to be a sensitive and accurate method for early diagnosis of EU-PRRSV infection.

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.

The new classification system for slaughter-pig herds in the Danish Salmonella surveillance-and-control program

The Danish surveillance-and-control program for Salmonella in slaughter pigs was introduced in 1995. The key element of the program is a quick and correct identification of herds with high seroprevalence. After 5 years, the classification scheme was evaluated—and a revision was made. Data from two Salmonella screenings including a total of 1902 slaughter pig herds were used. For each herd, information was available on Salmonella status based on both microbiology and serology. Based on analyses of these data, suitable changes in the scheme were identified and their effect estimated by use of data from the Danish Salmonella Database including all herds in 2000. The classification scheme has been adjusted on the following points. (1) The sampling has been simplified into 60, 75, or 100 samples per herd per year depending on herd size. This means more-precise estimates for the seroprevalence among smaller herds. (2) Herds with an annual kill ≤200 finishers will not form part of the surveillance; this leaves 1.6% of the slaughter pigs outside the surveillance scheme. (3) The cut-off for individual meat-juice samples has been reduced from OD% 40 to OD% 20—doubling the number of positive samples. (4) The results of the previous 3 months’ serological samples will be weighed 0.6:0.2:0.2 (the immediate month counting three times as much as the previous months), and the weighed average is called the “serological Salmonella index” for slaughter pig herds. A herd with an increasing seroprevalence will be assigned to a higher Salmonella level more-quickly under the new scheme. (5) A herd will be assigned monthly to one of three levels. The limit between Levels 1 and 2 has been set to ≥index 40, and the limit between Levels 2 and 3 to ≥index 70. If the Danish swine producers are interested, a Level 0 may be introduced (consisting of seronegative herds as an indication of a negligible Salmonella prevalence). The classification scheme was introduced in August 2001.

Salmonella in slaughter pigs: the effect of logistic slaughter procedures of pigs on the prevalence of Salmonella in pork

A substantial part of the finishing pigs in the Netherlands is infected with Salmonella. Infection of pigs with Salmonella can occur already on the farm. Pigs can also get infected or contaminated during transport, lairage or slaughter. The aim of this study was to evaluate the effect of separating pigs from Salmonella-infected farms from pigs from Salmonella-free farms during transport, lairage and slaughter on the prevalence of Salmonella on pork after slaughter. Two experiments were carried out. In the first experiment, farms were selected to participate, based on serology of the pigs (Dutch Salmonella ELISA). The pigs were slaughtered at the beginning of the day: firstly, sero-negative herds, secondly, sero-positive herds and thirdly, again sero-negative herds. The latter were slaughtered to investigate the effect of a contaminated slaughterline due to a previously slaughtered positive herd. In the second experiment, farms were selected to participate, based on both serology and bacteriology of the pigs on the farm. Two hundred pigs from Salmonella-free farms were slaughtered after 200 pigs from Salmonella-infected farms. Results showed that the prevalence of Salmonella in pork samples of sero-negative herds was lower than in samples of sero-positive herds. Results also showed that Salmonella contamination of carcasses after slaughter was partially caused by Salmonella-infected herds that were slaughtered before, and partially by residential flora of the slaughterhouse. It is concluded that separate slaughter of sero-negative pig herds can be useful to decrease the prevalence of Salmonella-contaminated pork after slaughter. To avoid cross-contamination by residential flora from trucks, lairage and slaughterline, cleaning and disinfection have to be improved.

Control of Yersinia enterocolitica in pigs at herd level

A higher herd prevalence of antibodies (ELISA) to Yersinia enterocolitica O:3 was found in conventional slaughter production (86.0% seropositive herds) than in conventional farrow-to-finish herds (53.1% seropositive herds). The herd prevalence of antibodies to Y. enterocolitica in multiplying herds (56.1%) was similar to the level in the conventional farrow-to-finish herds. An epidemiological study in conventional pig herds demonstrated that farrow-to-finish production (odds ratio, OR=0.15) was an important protective factor. Using under-pressure ventilation (OR=0.33) and manual feeding of slaughter pigs (OR=0.44) also lowered the herd prevalence. The most expressed risk factor was using an own farm vehicle for transport of slaughter pigs to abattoirs (OR=12.92). Separation between clean and unclean section in herds (OR=2.67), daily observations of a cat with kittens on the farm (OR=2.41) and using straw bedding for slaughter pigs (OR=2.25) were other factors that increased the risk. In conclusion, the epidemiological data suggest that it is possible to reduce the herd prevalence of Yersinia enterocolitica O:3 by minimising contact between infected herds and non-infected herds. Further, attempts to reduce the prevalence at the top levels of the breeding pyramids may be beneficial for the industry as a whole. The meat industry may use serological tests as a tool to lower the prevalence in the pig population by limiting the contact between seropositive and seronegative herds. However, because of the high prevalence of Y. enterocolitica in pig herds, a strict slaughter hygiene will remain an important means to reduce carcass contamination with Y. enterocolitica O:3 as well as other pathogenic micro-organisms.

Detection of brucellosis in diary herds after an outbreak of the disease using a delayed-type hypersensitivity test

A study of 14 herds was conducted to assess the skin delayed-type hypersensitivity (SDTH) test used in combination with the serum agglutination test and complement fixation test to curtail the spread of brucellosis. The tests were used to screen dairy herds suspected of brucellosis, and herds located near infected herds. The herds were tested every 5–6 weeks for 6 months. Cattle with a palpable skin swelling at the injection site of the allergen were considered as brucellosis suspects and were slaughtered. Supramammary lymph nodes of these cattle were collected and examined bacteriologically to determine their infection status. Brucellosis was detected and confirmed bacteriologically in three seronegative herds in which cattle tested positive with the SDTH test. Brucellosis was also confirmed bacteriologically in five seropositive herds in which cattle tested positive with the SDTH test. Brucellosis was not confirmed bacteriologically in one seronegative and four seropositive herds in which cattle reacted positively with the SDTH test. One herd reacted negatively in all tests. Because the SDTH test is able to detect latent carriers of Brucella organisms in seronegative herds, we recommend that the SDTH test be used in combination with serologic tests to curtail the spread of brucellosis.