Assessment of an Aujeszky's Disease Control Strategy in a Highly Prevalent Pig Farm Based on Systematic Vaccination With an Inactivated gE-Negative Marker Vaccine.

Aujeszky's disease (AD) is endemic in Argentina. In 2016, an inactivated gE- negative marker Bartha K61 vaccine (AUSKIPRA® BK) was launched for use, making Argentina the only country to carry out a control strategy plan with it. In the present article, we describe the results of a control program in a farrow-to-finishing farm with high initial AD prevalence (33% in sows), based on the systematic vaccination, detection, and elimination of seropositive pigs, the replacement of sows with vaccinated gilts, and the instauration of artificial insemination. The program was suitable for diminishing the incidence and the prevalence at levels consistent with virus eradication. This situation has been sustained over time. This is the first report of AUSKIPRA® BK efficacy under field conditions.

Reduction of foot-and-mouth disease virus transmission in cattle vaccinated one or two weeks before challenge using a commercial polyvalent vaccine.

Immediate vaccination of the most susceptible and epidemiological relevant animals is a crucial part of control measures that facilitate virus elimination in case of entry of foot-and-mouth disease (FMD). The objective of this study was to evaluate the effect of cattle vaccination 7 and 14 days prior challenge using a vaccine commonly applied in systematic vaccination campaigns against transmission of FMD virus (FMDV). Transmission of FMDV was investigated in three groups of ten cattle each: one non-vaccinated group and two groups that were either vaccinated 7 days (-7/vaccinated group) or 14 days (-14/vaccinated group) before intranasal (IN) inoculation. Five cattle heads from each group were inoculated using the IN-route with the A/Argentina/2001 FMDV strain, while the remaining five cattle heads of each group were contact-exposed to inoculated cattle. Clinical signs were recorded; virus isolation and genome detection by RT-PCR were carried out on oesophageal-pharyngeal fluid (OPF) and blood. Neutralizing antibody titers and antibodies against non-structural proteins (NSP) of FMDV were also determined. Results suggest that the experimental design, virus challenge dose, and virus infectivity were appropriate and that the virus had been transmitted to naïve calves. Under the outlined experimental conditions, vaccination 7 and 14 days prior to challenge induced full clinical protection against virus inoculation. Moreover, -7/ or -14/vaccinated calves that had been contact-exposed to -7/ or -14/vaccinated IN-challenged calves, did not become infected. Consequently, no virus transmission occurred from vaccinated and subsequently infected calves to cohabitating vaccinated calves (R = 0). According to our results, early vaccination during an outbreak is effective as virus transmission can be significantly reduced using a vaccine commercially available, routinely applied in systematic vaccination campaigns.

[Validation of a real time RT-PCR assay to detect foot-and-mouth disease virus and assessment of its performance in acute infection]. / Validación de una técnica de RT-PCR en tiempo real para la detección del virus de fiebre aftosa y evaluación de su desempeño en la infección aguda.

A specific real time reverse transcription polymerase chain reaction (RT-PCRrt) for the detection of foot-and-mouth disease virus was validated using the LightCycler thermocycler 2.0 and its reagents as recommended by the World Organization for Animal Health and was assessed for the detection of the virus in acute infection of cattle experimentally vaccinated and challenged with virus A Argentina/2001 or A24 Cruzeiro. The technique proved to be robust, showing coefficients of variation lower than 4% for different ARN extractions, days or repetitions and was able to detect up to 0,4 TCID 50%, and/or up to 100 RNA molecules. In probang samples, diagnostic sensitivity was 93.1 (95% CI 86.5-96.6) and diagnostic specificity 100 (95% CI 96.3-100). The results of the challenge in vaccinated or multivaccinated bovines showed that although there were high levels of clinical protection in the vaccinated group, FMDV could be detected in all challenged groups. However, detection was 100 times lower in immunized animals.

Validación de una técnica de RT-PCR en tiempo real para la detección del virus de fiebre aftosa y evaluación de su desempeño en la infección aguda. / [Validation of a real time RT-PCR assay to detect foot-and-mouth disease virus and assessment of its performance in acute infection].

A specific real time reverse transcription polymerase chain reaction (RT-PCRrt) for the detection of foot-and-mouth disease virus was validated using the LightCycler thermocycler 2.0 and its reagents as recommended by the World Organization for Animal Health and was assessed for the detection of the virus in acute infection of cattle experimentally vaccinated and challenged with virus A Argentina/2001 or A24 Cruzeiro. The technique proved to be robust, showing coefficients of variation lower than 4

for different ARN extractions, days or repetitions and was able to detect up to 0,4 TCID 50

, and/or up to 100 RNA molecules. In probang samples, diagnostic sensitivity was 93.1 (95

CI 86.5-96.6) and diagnostic specificity 100 (95

CI 96.3-100). The results of the challenge in vaccinated or multivaccinated bovines showed that although there were high levels of clinical protection in the vaccinated group, FMDV could be detected in all challenged groups. However, detection was 100 times lower in immunized animals.

Quantitative single serum-dilution liquid phase competitive blocking ELISA for the assessment of herd immunity and expected protection against foot-and-mouth disease virus in vaccinated cattle.

A single serum-dilution liquid phase ELISA (slpELISA) was standardized to be used for serological evaluation of herd immunity against foot-and-mouth disease. The absorbance value at a dilution 1:64 of each serum sample was interpolated in a standard curve by plotting the antibody titers of six control sera determined by end point dilution liquid phase ELISA (lpELISA), against the absorbance values for the same control sera at 1:64 dilutions. A straight line was obtained by linear regression analysis (r>0.90) in the titer range of 1.40-2.40. The reliability of the antibody titers was confirmed by the simultaneous titration of 60 cattle sera by slpELISA and lpELISA, which showed an acceptable correlation (R(2)>0.87) for viral strains A24/Cruzeiro, A/Argentina/01, O1/Campos and C3/Indaial. Titers obtained by both methods were not significantly different (p>0.05), thus confirming that slpELISA could be used successfully to replace the conventional serial dilution ELISA for the assessment of protection status of cattle in epidemiological studies. In addition, this quantitative slpELISA provides an adequate method for monitoring the effectiveness of vaccination campaigns and is also suitable for the assessment of seroconversion of naive animals during early stages of infection.

Reintroduction of foot-and-mouth disease in Argentina: characterisation of the isolates and development of tools for the control and eradication of the disease.

This paper describes the antigenic and molecular characterisation of foot-and-mouth disease virus (FMDV) strains isolated during the 2000-2002 epidemic in Argentina, and the strategy implemented for disease control. Two different FMDV serotypes, O and A, were involved. Of the various field isolates studied, two distinct O1 lineages (strains Corrientes/00 and Misiones/00) and two serotype A lineages (A/Argentina/00 and A/Argentina/01 prototypes) were identified. The genome sequences of these strains were compared with sequences of previous regional isolates and sequences of vaccine strains. O1 strains were found to be related to regional strains while serotype A strains were found to be more distanced from them. The updating of the antigenic composition of the vaccines used in the emergency was a key issue, since the outbreaks stopped shortly after the implementation of the vaccination programs. The O1 strains quickly disappeared from the field following strict control measures and the use of vaccines containing O1/Campos strain. However, in the case of the A serotype strains, the situation was different, since the use of a vaccine containing strain A24/Cruzeiro yielded acceptable levels of protection only after re-vaccination. Therefore, the new field strains A/Argentina/00 and A/Argentina/01 were incorporated into the vaccine, leading to an effective control of the disease. Viral circulation greatly diminished, as indicated by the significant reduction in the number of outbreaks and in the number of animals with antibodies against non-structural proteins. Satisfactory levels of protective antibodies were subsequently detected in the cattle population (above 75% protection). The absence of outbreaks after January 2002 indicated that the epidemic was controlled.