Antibody Response In Cattle Research Articles

Evaluation of commercial quadrivalent foot-and-mouth disease vaccines against east African virus strains reveals limited immunogenicity and duration of protection

Foot-and-mouth disease virus (FMDV) causes a contagious disease (FMD) in cloven-hoofed animals. For FMD-endemic countries, vaccination is critical for controlling disease but is rarely monitored, despite substantial funds spent on vaccine purchases. We evaluated antibody responses in cattle to two commercial vaccines each containing antigens of four FMDV serotypes. Sampling was done over 360 days, with serology for each serotype performed using commercially available solid phase competition ELISAs (SPCE) and with virus neutralization tests (VNT) employing regionally relevant test viruses. A primary course of each vaccine was administered to 37 calves, some of which received a second dose after 28 days. Using new production batches of vaccines, all calves received a booster vaccination 180 days post vaccination, while 10 additional naïve calves were also vaccinated using the new batches and followed up for ∼180 days. Simple and general linear models were used to compare antibody responses which varied substantially according to vaccine, dose regime, serotype, and test, but were mostly insufficient to ensure a high likelihood of adequate or sustained probable protection. One of the vaccines administered as a two-dose primary course of vaccination was superior to other options, but even then, data trajectories from VNT responses suggested probable protection of 75 % of calves for 6 months for only one virus serotype. Calves administered with the other vaccine and those given a single primary dose developed low levels of antibodies, offering predicted likely protection lasting less than two months. Individual SPCE results were weakly correlated (r2 = 0.48) to neutralization and associated likelihoods of protection but SPCE and VNT agreed on which vaccine and dose regime performed best. Our findings highlight gaps in immunogenicity of FMD vaccines used in East Africa and reinforce the importance of independent quality control studies to evaluate and improve commercial FMD vaccines and vaccination regimes.

Antibody response in cattle after local isolate SE vaccine administration

Septicemic Epizootica (SE) in livestock caused by bacterial infection of Pasteurella multocida. The serotype of P. multocida in Indonesia is the Asian serotype B:2. The objective of this study is to determine antibody responses of cattle after vaccinating with local isolates Local isolate SE Vaccine. Ten female cattle aged around one year used in this experiment. Cattle divided into 3 groups, Group A (n=4) vaccinated with local isolates SE vaccines, Group B (n=4) vaccinated with commercial SE vaccines and Group C (n=2) unvaccinated. The concentration of SE vaccine is 2mg, and the vaccine was administrated 3ml subcutaneously. Antibody responses measured before and after vaccination using indirect-ELISA. The result of antibody titre of cattle vaccinated with local isolate SE vaccine in seppic-montanide formulation showed higher results compared to SE commercial vaccine and unvaccinated group. This local isolate SE vaccine has good potential as a candidate for SE vaccine in cattle.

Effect of Phage Targeting Therapy of Brucellosis on Host Antibody Response in Cattle.

Brucellosis caused by Brucella organisms is a major zoonosis globally. It causes heavy losses through abortions, delayed conception, and infertility in animals. Brucella is an intracellular bacterium. Antibiotic therapy for bovine Brucellosis is expensive and may sometimes be ineffective as Brucella can become resistant. Once infected, the animal may remain carrier and shed bacteria in milk, semen, and uterine discharges spreading infection to others for a long period. The live attenuated Brucella abortus strain S19 organisms that are commonly used as a vaccine were employed to deliver a broad acting lytic brucellaphage inside the phagocytes in vivo to reach the virulent Brucella hiding intracellularly. The phage pulsed vaccine induced sustained and significantly high titers of anti-Brucella antibodies compared with the untreated animals and animals vaccinated with S19 vaccine alone as estimated by standard tube agglutination test, microagglutination test, indirect hemagglutination assay test, and enzyme-linked immunosorbent assay. The current investigation is perhaps the first systematic attempt whereby efficacy of brucellaphage pulsed vaccine preparation in induction of specific antibody response was evaluated in cattle.

Molecular Dissection of the Antibody Response: Opportunities and Needs for Application in Cattle.

Improving understanding of the bovine adaptive immune response would equip researchers to more efficiently design interventions against pathogens that impact upon food security and animal welfare. There are features of the bovine antibody response that differ substantially from other mammalian species, including the best understood models in the human and mouse. These include the ability to generate a functionally diverse immunoglobulin response despite having a fraction of the germline gene diversity that underpins this process in humans and mice, and the unique structure of a subset of immunoglobulins with “ultralong” HCDR3 domains, which are of significant interest with respect to potential therapeutics, including against human pathogens. However, a more detailed understanding of the B cell response and the production of an effective antibody response in the bovine is currently hampered by the lack of reagents for the B cell lineage. In this article we outline the current state of knowledge and capabilities with regard to B cell and antibody responses in cattle, highlight resource gaps, and summarize recent advances that have the potential to fundamentally advance our understanding of this process in the bovine host.

Exploring the longitudinal dynamics of herd BVD antibody test results using model-based clustering

Determining the Bovine Viral Diarrhoea (BVD) infection status of cattle herds is a challenge for control and eradication schemes. Given the changing dynamics of BVD virus (BVDV) antibody responses in cattle, classifying herds based on longitudinal changes in the results of BVDV antibody tests could offer a novel, complementary approach to categorising herds that is less likely than the present system to result in a herd’s status changing from year to year, as it is more likely to capture the true exposure dynamics of the farms. This paper describes the dynamics of BVDV antibody test values (measured as percentage positivity (PP)) obtained from 15,500 bovines between 2007 and 2010 from thirty nine cattle herds located in Scotland and Northern England. It explores approaches of classifying herds based on trend, magnitude and shape of their antibody PP trajectories and investigates the epidemiological similarities between farms within the same cluster. Gaussian mixture models were used for the magnitude and shape clustering. Epidemiologically meaningful clusters were obtained. Farm cluster membership depends on clustering approach used. Moderate concordance was found between the shape and magnitude clusters. These methods hold potential for application to enhance control efforts for BVD and other infectious livestock diseases.

Molecular vaccine prepared by fusion of XCL1 to the multi-epitope protein of foot-and-mouth disease virus enhances the specific humoural immune response in cattle.

Targeting antigen to dendritic cells (DCs) is a promising way to manipulate the immune response and to design prophylactic molecular vaccines. In this study, the cattle XCL1, ligand of XCR1, was fused to the type O foot-and-mouth disease virus (FMDV) multi-epitope protein (XCL-OB7) to create a molecular vaccine antigen, and an △XCL-OB7 protein with a mutation in XCL1 was used as the control. XCL-OB7 protein specifically bound to the XCR1 receptor, as detected by flow cytometry. Cattle vaccinated with XCL-OB7 showed a significantly higher antibody response than that to the △XCL-OB7 control (P<0.05). In contrast, when XCL-OB7 was incorporated with poly (I:C) to prepare the vaccine, the antibody response of the immunized cattle was significantly decreased in this group and was lower than that in the △XCL-OB7 plus poly (I:C) group. The FMDV challenge indicated that cattle immunized with the XCL-OB7 alone or the △XCL-OB7 plus poly (I:C) obtained an 80% (4/5) clinical protective rate. However, cattle vaccinated with △XCL-OB7 plus poly (I:C) showed more effective inhibition of virus replication than that in the XCL-OB7 group after viral challenge, according to the presence of antibodies against FMDV non-structural protein 3B. This is the first test of DC-targeted vaccines in veterinary medicine to use XCL1 fused to FMDV antigens. This primary result showed that an XCL1-based molecular vaccine enhanced the antibody response in cattle. This knowledge should be valuable for the development of antibody-dependent vaccines for some infectious diseases in cattle.

Recombinant human adenovirus-5 expressing capsid proteins of Indian vaccine strains of foot-and-mouth disease virus elicits effective antibody response in cattle

Recombinant adenovirus-5 vectored foot-and-mouth disease constructs (Ad5- FMD) were made for three Indian vaccine virus serotypes O, A and Asia 1. Constructs co-expressing foot-and- mouth disease virus (FMDV) capsid and viral 3C protease sequences, were evaluated for their ability to induce a neutralizing antibody response in indigenous cattle (Bos indicus). Purified Ad5-FMD viruses were inoculated in cattle as monovalent (5×109 pfu/animal) or trivalent (5×109 pfu/animal per serotype) vaccines. Animals vaccinated with monovalent Ad5-FMD vaccines were boosted 63days later with the same dose. After primary immunization, virus neutralization tests (VNT) showed seroconversion in 83, 67 and 33% of animals vaccinated with Ad5-FMD O, A and Asia 1, respectively. Booster immunization elicited seroconversion in all of the animals (100%) in the monovalent groups. When used in a trivalent form, the Ad5-FMD vaccine induced neutralizing antibodies in only 33, 50 and 16% of animals against serotypes O, A and Asia 1, respectively on primo-vaccination, and titers were significantly lower than when the same vectors were used in monovalent form. Neutralizing antibody titers differed by serotype for both Ad5-FMD monovalent and trivalent vaccines, with Asia 1 serotype inducing the lowest titers. Antibody response to Ad5 vector in immunized cattle was also assessed by VNT. It appeared that the vector immunity did not impact the recall responses to expressed FMDV antigens on booster immunization. In summary, the study suggested that the recombinant Ad5-FMD vaccine has a potential use in monovalent form, while its application in multivalent form is not currently encouraging.

The efficacy of alcelaphine herpesvirus-1 (AlHV-1) immunization with the adjuvants Emulsigen® and the monomeric TLR5 ligand FliC in zebu cattle against AlHV-1 malignant catarrhal fever induced by experimental virus challenge

Malignant catarrhal fever (MCF) is a fatal disease of cattle that, in East Africa, follows contact with wildebeest excreting alcelaphine herpesvirus 1 (AlHV-1). Recently an attenuated vaccine (atAlHV-1) was tested under experimental challenge on Friesian-Holstein (FH) cattle and gave a vaccine efficacy (VE) of approximately 90%. However testing under field conditions on an East African breed, the shorthorn zebu cross (SZC), gave a VE of 56% suggesting that FH and SZC cattle may respond differently to the vaccine. To investigate, a challenge trial was carried out using SZC. Additionally three adjuvant combinations were tested: (i) Emulsigen®, (ii) bacterial flagellin (FliC) and (iii) Emulsigen®+bacterial flagellin. We report 100% seroconversion in all immunized cattle. The group inoculated with atAlHV-1+Emulsigen® had significantly higher antibody titres than groups inoculated with FliC, the smallest number of animals that became infected and the fewest fatalities, suggesting this was the most effective combination. A larger study is required to more accurately determine the protective effect of this regime in SZC. There was an apparent inhibition of the antibody response in cattle inoculated with atAlHV-1+FliC, suggesting FliC might induce an immune suppressive mechanism. The VE in SZC (50–60%) was less than that in FH (80–90%). We speculate that this might be due to increased risk of disease in vaccinated SZC (suggesting that the vaccine may be less effective at stimulating an appropriate immune response in this breed) and/or increased survival in unvaccinated SZC (suggesting that these cattle may have a degree of prior immunity against infection with AlHV-1).

A comparison of titers of anti-Brucella antibodies of naturally infected and healthy vaccinated cattle by standard tube agglutination test, microtiter plate agglutination test, indirect hemagglutination assay, and indirect enzyme-linked immunosorbent assay.

Aim:We determined the antibody response in cattle naturally infected with brucellosis and normal healthy adult cattle vaccinated during calf hood with strain 19.Materials and Methods:The antibody titers were measured by standard tube agglutination test (STAT), microtiter plate agglutination test (MAT), indirect hemagglutination assay (IHA), and indirect enzyme-linked immunosorbent assay (iELISA) as per standard protocols.Results:The mean STAT titers were 1.963±0.345 in infected cattle and 1.200±0.155 in healthy vaccinated cattle. The difference was extremely significant (p<0.0001). The mean MAT titers were 2.244±0.727 in infected cattle and 1.200±0.155 in healthy vaccinated cattle. The difference was very significant (p<0.005). The mean IHA titers in infected cattle were 2.284±0.574, and those in healthy vaccinated cattle were 1.200±0.155. The difference was extremely significant (p=0.0002). However, the difference in mean iELISA titers of infected cattle (1.3678±0.014) and healthy vaccinated cattle (1.367±0.014) was non-significant. The infected animals showed very high titers of agglutinating antibodies compared to the vaccinated animals. However, it cannot be ascertained whether these antibodies are due to vaccine or response to infection. Since the infected animals had been vaccinated earlier, the current infection may suggest that vaccination was unable to induce protective levels of antibody. The heightened antibody response after infection may also indicate a secondary immune response to the antigens common to the vaccine strain and wild Brucella organisms.Conclusion:The brucellosis infected animals showed very high titers of agglutinating antibodies compared to the vaccinated animals.

No significant differences in the breadth of the foot-and-mouth disease serotype A vaccine induced antibody responses in cattle, using different adjuvants, mixed antigens and different routes of administration

Inactivated whole virus foot-and-mouth disease (FMD) vaccines are used worldwide for protection against FMD, but not all vaccines induce protection against all genetic variants of the same FMD virus serotype. The aim of this study is to investigate whether the “breadth” of the antibody response against different strains of the same FMD virus serotype in cattle could be improved by using a different adjuvant, a mix of antigens and/or different routes of administration. To this end, six groups of five cattle were vaccinated with different FMD virus serotype A strain vaccines formulated with Montanide ISA 206 VG adjuvant. Antibody responses for homologous and heterologous cross-reactivity against a panel of 10 different FMD virus serotype A strains were tested by a liquid-phase blocking ELISA. Results of cattle vaccinated with ISA 206 VG adjuvanted vaccine were compared with results obtained in a previous study using aluminium hydroxide-saponin adjuvant. No significant effect of adjuvant on the breadth of the antibody response was observed, neither for mixing of antigens nor for the route of administration (subcutaneous vs. intradermal). Comparison of antigen payload, however, increased both homologous and heterologous titres; a 10-fold higher antigen dose resulted in approximately four times higher titres against all tested strains. Our study shows that breadth of the antibody response depends mainly on the vaccine strain; we therefore propose that, for vaccine preparation, only FMD virus strains are selected that, among other important characteristics, will induce a wide antibody response to different field strains.

Morphological and immunological characterization of anthrax vaccine in cattle

Anthrax is a zoonotic disease caused by the bacterium Bacillus anthracis that normally affects animals, especially ruminants (such as cattle, goats, sheep, and horses) and humans. This study was planned to characterize the morphology of anthrax vaccine bacteria by using Gram’s stain, polychrome methylene blue stain, culture on nutrient agar and nutrient broth media and to determine the immunological status of this vaccine by indirect ELISA and slide agglutination test in cattle. Antibiotic sensitivity test of vaccine strain of bacteria was also done. This study provided evidence that vaccine strain of Bacillus anthracis was gram-positive rod-shaped bacteria appeared as single to short-chained bacilli with blunted ends. Serum from anthrax vaccinated cattle agglutinated anthrax antigen on Day 30 (+++ within 5 min and ++ within 7 min at 1:100 dilution of test sera) and Day 90 (+++ within 5 min at 1:100 dilution of test sera) of post immunization. Immunization of cattle with anthrax vaccine generated high level of anti-anthrax IgG antibody response at Day 30 (0.789}0.014) of post immunization and reached its peak at Day 90 (0.991}0.047). This study also provided evidence that anthrax vaccine bacteria were sensitive to penicillin, streptomycin, amoxicillin and kenamycin. It may be recommended that the anthrax vaccine currently in use in Bangladesh is proved to be effective in term of morphology of Bacillus antharis and raising anti anthrax IgG antibody response in cattle with no side effect. DOI: http://dx.doi.org/10.3329/bjvm.v11i1.17732 Bangl. J. Vet. Med. (2013). 11 (1): 43-49

Antibody Response in Cattle and Guinea Pigs Inoculated with Rabies Vaccines

One hundred ninety-five rabies cases in cattle were identified in South Korea since 1993. As most of rabies cases have a relation to rabid Korean raccoon dogs (Nyctereutes procyonoides koreensis), vaccination to animals including cattle is mandatory in rabies risk region. In order to minimize fatal rabies in animals, eradication policy of the disease has been achieved by controlling reservoirs and by mass vaccination. In this study, we compared the antibody response in cattle and guinea pigs inoculated with rabies vaccines commercially available in Korea. Each group of cattle in Gangwon-do was vaccinated intramuscularly with either one of five commercial inactivated vaccines or a live attenuated rabies vaccine (designated as A to F). Serum samples at the time of vaccination and four weeks post vaccination were obtained from the cattle and guinea pigs and were analyzed with virus neutralizing assay (VNA). Each group of cattle inoculating rabies vaccines showed significant virus neutralizing antibody titers (p < 0.05) ranging from 1.55 to 17.8 mean IU/ml compared with the non-vaccinated cattle and guinea pigs inoculated with 1/20 dose of vaccine showed relatively low VN antibody titers ranging from 0.23 to 6.1 mean IU/ml. All cattle immunized with A, C and F showed high VN antibody titers over 0.5 IU/ml and 62.5% and 37.5% of cattle inoculated with D and E showed protective antibody titer, respectively. This finding suggests that the inactivated or live attenuated rabies vaccination commercially available in Korea could induce protective antibody response in Korean cattle, but sero-conversion rate and sero-positive rate showing VN antibody titer over 0.5 IU/ml depend on vaccines.

A One-year Follow-up of Antibody Response in Cattle and Sheep after Vaccination with Serotype 8- and Serotype 1-inactivated BT Vaccines

Sixteen sheep and 18 cattle were followed up during 1year to estimate the duration of immunity induced by inactivated bluetongue virus serotype 8 (BTV-8) vaccines (sheep and cattle) and a bluetongue virus serotype 1 (BTV-1) vaccine (cattle) under field conditions using cELISA and seroneutralization test (SNT). Four sheep never seroconverted. Those that seroconverted were all seronegative by BTV-8 SNT at the date of last sampling [378days post-vaccination (dpv)]. Eight sheep were still positive by competitive ELISA (cELISA) 378 dpv. All the cattle seroconverted. At the end of the study, eight and 11 cattle were still positive by BTV-8 SNT and cELISA, respectively (335dpv); and nine were still positive by BTV-1 SNT (301dpv).

Avidity and subtyping of specific antibodies applied to the indirect assessment of heterologous protection against Foot-and-Mouth Disease Virus in cattle

Serological assessment of the heterologous response among Foot-and-Mouth Disease Virus (FMDV) strains is mainly performed by virus neutralization test (VNT), liquid phase blocking ELISA and complement fixation assay. In this study two high-throughput ELISA techniques, avidity and IgG subtype ELISA, were developed and used to further characterize heterologous antibody responses in cattle during vaccination and challenge. Both assays were applied to a set of previously characterized sera from animals immunized with an inactivated A24 Cruzeiro/Brazil/55 (A24 Cruzeiro) strain monovalent FMDV vaccine and challenged with the heterologous A/Argentina/2001 (A/Arg/01) strain. Single dilution avidity ELISA assessment showed that animals that were protected against A/Arg/01 challenge had higher avidity antibodies to this heterologous strain than non-protected cattle. Animals with low or even undetectable anti-A/Arg/01 serum-neutralizing titers that passed the heterologous challenge presented higher IgG1/IgG2 ratio than non-protected animals. In this study, the three assessments (VNT and both ELISAs) discriminated between protected and not protected animals against a heterologous challenge. The combination of these techniques may be applied to complement current indirect serological vaccine-matching assessments. The measurement of these qualitative parameters may provide additional information to understand the mechanisms underlying FMD heterologous responses and the induction of cross-protection in cattle.

Duration of protective immunity and antibody responses in cattle immunised against alcelaphine herpesvirus-1-induced malignant catarrhal fever.

Protection of cattle from alcelaphine herpesvirus-1 (AlHV-1)-induced malignant catarrhal fever (MCF) has been described previously, using an attenuated virus vaccine in an unlicensed adjuvant. The vaccine was hypothesised to induce a protective barrier of virus-neutralising antibody in the oro-nasal region, supported by the observation of high titre neutralising antibodies in nasal secretions of protected animals. Here we describe further analysis of this vaccine strategy, studying the effectiveness of the vaccine formulated with a licensed adjuvant; the duration of immunity induced; and the virus-specific antibody responses in plasma and nasal secretions. The results presented here show that the attenuated AlHV-1 vaccine in a licensed adjuvant protected cattle from fatal intranasal challenge with pathogenic AlHV-1 at three or six months. In addition, animals protected from MCF had significantly higher initial anti-viral antibody titres than animals that succumbed to disease; and these antibody titres remained relatively stable after challenge, while titres in vaccinated animals with MCF increased significantly prior to the onset of clinical disease. These data support the view that a mucosal barrier of neutralising antibody blocks infection of vaccinated animals and suggests that the magnitude of the initial response may correlate with long-term protection. Interestingly, the high titre virus-neutralising antibody responses seen in animals that succumbed to MCF after vaccination were not protective.

Cross immunity with Haemaphysalis longicornis glutathione S-transferase reduces an experimental Rhipicephalus (Boophilus) microplus infestation

Recombinant Glutathione S-transferase of Haemaphysalis longicornis (rGST-Hl) was expressed in Escherichia coli, purified by affinity chromatography and used in the immunization of cattle. Western blot analysis showed positive antibody response in cattle immunized with rGST-Hl. The tests also showed that immunized bovine sera recognize native Rhipicephalus microplus proteins in different tissue extracts. Furthermore, the vaccine potential of rGST-Hl was investigated against infestation of Hereford cattle by R. microplus. Vaccination of cattle with rGST-Hl conferred partial cross-protection immunity against R. microplus. Considering the effect on number of engorged ticks, egg laying capacity and egg fertility, the overall efficacy of vaccination was of 57%, as compared with control group.

Neutralising antibody responses in cattle and sheep following booster vaccination with two commercial inactivated bluetongue virus serotype 8 vaccines

Cattle and sheep that had received a primary course of vaccination with an inactivated bluetongue virus serotype 8 (BTV-8) vaccine were booster vaccinated 6 or 12 months later with the homologous vaccine or an alternative inactivated BTV-8 vaccine and neutralising antibody responses were determined. Antibody titres to the alternative vaccine were significantly higher than to the homologous vaccine (P=0.013) in cattle. There was no significant difference between the antibody responses to alternative and homologous vaccines in sheep. These data indicate that cattle and sheep primed with one inactivated BTV-8 vaccine may be effectively boosted with an alternative commercial inactivated BTV-8 vaccine.

Antibody Response in Cattle Vaccinated Against Bluetongue Serotype 8 in Italy

To assess the immunogenicity of Zulvac 8 Bovis (a commercial inactivated vaccine against bluetongue virus serotype 8 - BTV8) under field conditions, 71 cattle vaccinated according to manufacturer schedule in Verona province (Italy) were tested for the presence of BTV8 neutralizing antibodies at 21, 29, 36, 43, 49, 102 and 201 days post-vaccination (dpv). Another group of 528 BTV8 vaccinated cattle in Mantova province (Italy) was also tested once between 113 and 174 dpv. The vaccine was able to elicit an immune response in 69 (97.2%) and 346 (65.5%) animals of the Verona and Mantova groups, respectively.

Simple methods for measurement of bovine mucosal antibody responses in vivo

The mucosal immune response serves as the first line of defence against many bacterial and viral diseases. Therefore, measurement of mucosal immune responses is important in evaluating mucosal immunisation protocols and understanding initial host/pathogen interactions. In this study we compare two methods for repeated sampling of bovine rectal mucosal secretions, namely rectal swabbing and rectal biopsies, and evaluate a simple swabbing method for sampling bovine nasal secretions. Both rectal swabs and rectal biopsies yielded similar quantities of total IgA (TIgA)/ml. However, rectal biopsies yielded five times more total IgG (TIgG)/ml than rectal swabs. Blood contamination was estimated to contribute approximately 7% of TIgG and <0.05% TIgA in rectal swab samples compared to 40% of TIgG and 4.5% of TIgA in rectal biopsy samples, indicating that rectal swabbing was more effective at sampling rectal mucosal secretions. Nasal swabs were effective at obtaining nasal secretion samples with only 1% of TIgG and <0.05% TIgA estimated to be blood derived. Furthermore, H7 flagellin-specific antibodies were detected in both nasal and rectal swab samples following either rectal immunisation with purified H7 flagellin or oral challenge with live E. coli O157:H7, indicating that both techniques are effective methods for monitoring mucosal antibody responses in cattle.

Use of an immunodominant p17 antigenic fraction of Neospora caninum in detection of antibody response in cattle

A Neospora caninum 17 kDa protein fraction (p17) has been described as an immunodominant antigen (IDA) under reducing and non-reducing conditions. The aim of the present study was to investigate the diagnostic utility of p17 in cattle. In order to achieve this, p17 was purified by electroelution from whole N. caninum tachyzoite soluble extract and a p17-based Western blot (WB-p17) was developed. The p17 recognition was measured by densitometry and expressed as OD values to check the validity of the WB-p17. A total of 131 sera including sequential samples from naturally- and experimentally-infected calves and breeding cattle were analysed by WB-p17 and compared with IFAT using whole formalin-fixed tachyzoites as a reference test. The results obtained highlight the feasibility of using the N. caninum p17 in a diagnostic test in cattle. Firstly, the assay based on the p-17 antigen discriminated between known positive and negative sera from different cattle populations, breeding cattle and calves. Secondly, the p17 antigen detected fluctuations in the antibody levels and seroconversion in naturally- and experimentally-infected cattle. Significant differences in p-17 antigen recognition were observed between naturally infected aborting and non-aborting cattle, as well as significant antibody fluctuations over time in experimentally infected cattle, which varied between groups. Furthermore, the results obtained with WB-p17 are in accordance with the results obtained with the IFAT as high agreement values were obtained when all bovine subpopulations were included (kappa = 0.86).