The first isolation of equine arteritis virus in Argentina

Echeverría, María Gabriela; Pecoraro, Marcelo Ricardo Ítalo; Galosi, Cecília Mônica; Etcheverrigaray, María Elisa; Nosetto, Edgardo Omar

doi:10.20506/rst.22.3.1458

Cited by 21 publications

(20 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Positive RT-PCR fi nding of EAV in the semen of breeding stallions is the key evidence for the presence of EVA, which has been confi rmed in the reports of numerous researchers [11,14,15]. In our study, EAV was detected by RT-PCR in all (11) tested semen samples from one (stallion № 1) of the three examined stallions, during the mating season in 2013 and 2014.…”

Section: Discussionsupporting

confidence: 85%

See 1 more Smart Citation

Detection Of Equine Arteritis Virus In The Semen Of Stallions In The Republic Of Serbia

et al. 2015

View full text Add to dashboard Cite

The results on serological testing of blood sera from stallions and mares used for breeding and the presence of the viral genome of Equine Arteritis Virus (EAV) in stallion semen are presented. The blood and semen samples were taken from a horse stable on the territory of the Republic of Serbia during 2012, 2013 and 2014. Detection of anti-EAV specifi c antibodies in blood sera was performed by the virus neutralization test (VNT), and identifi cation of EAV genome RNA in stallion semen was done by reverse-transcriptase polymerase chain reaction (RT-PCR). In 2012, high seroprevalence of EAV was detected in the investigated stable. In total, 45% and 65 % of stallions and mares reacted positive, respectively, and the antibody titre values ranged between 2 and 10 log 2 . High seroprevalence was confi rmed in the same animals again in 2013. Out of two stallions tested semen samples in 2013, the viral genome was detected by RT-PCR in 3 examined semen samples from a seropositive stallion, while EAV was not detected in 3 semen samples of a seronegative stallion. During 2014, 11 semen samples were collected from two seropositive stallions. Again, the presence of EAV was confi rmed by RT-PCR in all 8 semen samples originating from the same stallion with the EAV genome positive semen result in 2013, whereas the virus was not detected in semen samples originating from the second anti-EAV antibody positive stallion. The presence of EAV-specifi c antibodies was confi rmed in the blood sera of the mares inseminated with the semen of seropositive stallions in 2012 and 2013.

show abstract

Section: Discussionsupporting

confidence: 85%

“…A certain attention in laboratory testing should be paid to stallions. During the past 20 years, different molecular methods have been developed with the aim of a rapid and effective virus detection in stallion semen, based on reverse transcriptase-polymerase chain reaction (RT-PCR) [6,8,11,15].…”

Section: Introductionmentioning

confidence: 99%

Detection Of Equine Arteritis Virus In The Semen Of Stallions In The Republic Of Serbia

et al. 2015

View full text Add to dashboard Cite

show abstract

“…Before castration of the animal, a semen sample was processed for routine virus isolation at the Virology Laboratory, Faculty of Veterinary Sciences, National University of La Plata, as described previously (3,17). Briefly, serial tenfold dilutions (10 -1 to 10 -3 ) of the sample were inoculated in duplicate onto RK13 cells grown in six-well plates.…”

Section: Sample Processing and Virological Methodologymentioning

confidence: 99%

“…The phylogenetic dataset was built with five strains isolated in Argentina, sequences reported previously (3,4,5,6,11), and a group of selected sequences obtained from GenBank (Table I). Sequences were edited using BioEdit software version 5 (19) and aligned in molecular evolutionary genetic analysis (MEGA) software, version 4.0, using the ClustalW algorithm.…”

Section: Dataset and Phylogenetic Analysismentioning

confidence: 99%

The equine arteritis virus isolate from the 2010 Argentinian outbreak

Metz¹,

Serena²,

Panei³

et al. 2014

Rev. Sci. Tech. OIE

Self Cite

View full text Add to dashboard Cite

SummaryA semen sample from a stallion infected during the 2010 equine arteritis virus (EAV) outbreak was received for viral isolation prior to castration of the animal. The virus was identified using a polyclonal antibody immunofluorescence test. Reverse-transcription polymerase chain reaction (RT-PCR) was used to amplify a region of the GP5 gene with primers GL105F and GL673R. The PCR products were purified and sequences of both strands were determined in a MegaBACE ™ 1000 with inner primers CR2 and EAV32. A phylogenetic dataset was built with the previously reported sequences of five strains isolated in Argentina, together with a group of selected sequences obtained from GenBank. The unrooted neighbourjoining tree was constructed using molecular evolutionary genetic analysis (MEGA) and bootstrap analyses were conducted using 1,000 replicate datasets. Evolutionary distances were computed using the maximum composite likelihood method. A NetNGlyc server analysis at the Technical University of Denmark (www.cbs.dtu.dk/services/NetNGlyc/) was used to predict N-glycosylation in GP5 sequences. The phylogenetic analysis revealed that the new strain (GLD-LP-ARG), together with other strains previously isolated, belongs to the European group EU-1 but in a different branch. The new strain shows 99% nucleotide identity with strain A1 and 98.1% with the Belgian strain 08P178. Persistently infected stallions and their cryopreserved semen constitute a reservoir of EAV, which ensures its persistence in the horse population around the world. These findings reinforce the importance of careful monitoring of persistently infected stallions, as well as semen straws, by RT-PCR or test mating, in accordance with national regulations.

show abstract

“…However, the most important clinical signs 9 of the disease are abortions, respiratory disease in adult animals and pneumonia in foals. 10In Argentina, although serological evidence was first documented in 1984 (Nosetto et al, 11 1984), EAV was first isolated in 2001(Echeverría et al, 2003. Following the EAV 12 outbreak in 2010, the number of samples sent to the laboratory for EAV analysis was 13 significantly higher than the annual average of the previous years, reaching almost 5000 14 samples analyzed over a period of seven months.…”

mentioning

confidence: 99%

Development of a peptide ELISA for the diagnosis of Equine arteritis virus

Metz

Lorenzón

Serena

et al. 2014

Journal of Virological Methods

View full text Add to dashboard Cite

19A peptide-based indirect ELISA was developed to detect antibodies against Equine 20 arteritis virus (EAV). Two peptides for epitope C of protein GP5 and fragment E of protein 21 M were designed, synthesized, purified and used as antigens either alone or combined. 22Ninety-two serum samples obtained from the 2010 equine viral arteritis outbreak, analyzed 23 previously by virus neutralization, were evaluated by the ELISA here developed. The best 24 resolution was obtained using peptide GP5. The analysis of the inter-and intraplate 25 variability showed that the assay was robust. The results allow concluding that this 26Page 2 of 16 A c c e p t e d M a n u s c r i p t 2 peptide-based ELISA is a good alternative to the OIE-prescribed virus neutralization test 1 because it can be standardized between laboratories, can serve as rapid screening, can 2 improve the speed of diagnosis of EAV-negative horses and can be particularly useful for 3 routine surveillance in large populations. 4 KEY WORDS synthetic peptides-ELISA-Equine arteritis virus 5 6 Equine arteritis virus (EAV) belongs to the order Nidovirales, family Arteriviridae, 7 genus Arterivirus (Snijder and Meulenberg, 1998). The most relevant feature of EAV 8 infection is that it produces subclinical infection. However, the most important clinical signs 9 of the disease are abortions, respiratory disease in adult animals and pneumonia in foals. 10In Argentina, although serological evidence was first documented in 1984 (Nosetto et al., 11 1984), EAV was first isolated in 2001(Echeverría et al., 2003. Following the EAV 12 outbreak in 2010, the number of samples sent to the laboratory for EAV analysis was 13 significantly higher than the annual average of the previous years, reaching almost 5000 14 samples analyzed over a period of seven months. This increase highlighted the need for 15 an alternative technique to replace the virus neutralization test, which is, to date, the test 16 for international trade prescribed by the OIE (OIE, 2012). The virus neutralization test 17 detects antibodies against to EAV GP5 protein but is complex and high cost and requires 18 72 h to yield a result. Other difficulties include the considerable interlaboratory variation 19 and the contamination or nonspecific cellular cytotoxicity in sera from vaccinated horses 20 (Newton et al., 2004). Although several ELISAs have been developed, none have been 21 validated as extensively as the virus neutralization test. Some, however, offer comparable 22 specificity and almost equivalent sensitivity. The aim of this work was to design an ELISA 23 as a screening assay for EAV, using synthetic peptides. Indirect ELISA using peptides 24 containing GP5 neutralization epitopes may provide a simpler and more cost-effective 25 method to quantify EAV antibodies than the virus neutralization test. Another benefit of an 26Page 3 of 16 A c c e p t e d M a n u s c r i p t 3 EAV ELISA is that it can provide a same-day test result compared with the 72 h needed for 1 the virus neutralization test....

show abstract

The first isolation of equine arteritis virus in Argentina

Cited by 21 publications

References 8 publications

Detection Of Equine Arteritis Virus In The Semen Of Stallions In The Republic Of Serbia

Detection Of Equine Arteritis Virus In The Semen Of Stallions In The Republic Of Serbia

The equine arteritis virus isolate from the 2010 Argentinian outbreak

Development of a peptide ELISA for the diagnosis of Equine arteritis virus

Contact Info

Product

Resources

About