AGID Test Research Articles

Field Study of Infectious Bursal Disease in A Laying Poultry Farm at Khartoum State, Sudan

The aim of this study is to investigate field Infectious bursal disease outbreak in a laying poultry farm. The disease is an economically important worldwide disease in poultry production sector. Serological tests (Elisa, AGID, and immunochromatography), morbidity rate, mortality rate and post-mortem changes were studied. The results showed 100% morbidity, 28% mortality, positive AGID test and immunochromatography test and the Elisa test which was done at the end of the outbreak. Post-mortem revealed haemorrhages in the enlarged and edematous burasa, atrophy of the bursa, haemorrhage in skin, breast, thigh muscles and proventriculus as the most frequently detected lesions. The kidneys were affected showing enlargement, nut meg appearance and pale color Deposition of urates in ureters was also obvious. In conclusion IBD is a serious problem causing a considerable socioeconomic loss for the farmer, consumers and the country as a whole. The disease can be diagnosed using serological tests. IBD needs extensive effort in extension to raise the awareness of the sector about the disease diagnosis, prevention and control.

Identificação genética, aspectos clínicos e epidemiológicos de um foco de anemia infecciosa equina no Estado do Rio Grande do Sul, Brasil

ABSTRACT: The study describes the genetic identification, clinical, and epidemiological characteristics of an outbreak of equine infectious anemia occurring in the state of Rio Grande do Sul, Brazil. Three animals kept in the periurban region of Uruguaiana city tested positive for the AGID test. The serology was performed as a requirement for transit. None of the animals showed clinical signs of infection, one animal was necropsied, and the others were stolen. In the post-mortem examination, no macroscopic changes were observed, and microscopically, discrete hemosiderosis was detected in fragments of the liver and spleen. Amplifying and sequencing a proviral DNA fragment in blood, spleen, and mesenteric lymph node samples confirmed EIAV infection. Phylogenetic analysis of the first sequenced EIAV sample from the Rio Grande do Sul State indicates a high similarity with other Brazilian samples. Results confirmed the viral presence in the state’s herds and described epidemiological and virological characteristics of EIA that contribute to the maintenance and dissemination of the virus in herds.

Impacto econômico e aspectos epidemiológicos da mastite e artrite encefalite caprina na região semiárida do Nordeste Brasileiro

Dairy activity in the semi-arid region has gained prominence due to its profit potential; however, there are still sanitary problems that result in significant economic impact. Thus, this study aimed to assess the economic impact and to conduct an epidemiological study of caprine arthritis-encephalitis (CAE) and subclinical mastitis in dairy goat herds of the semi-arid northeastern region of Brazil. The study proceeded with the collection of blood (n = 351) and milk samples (n = 321) from 15 rural properties with up to 50 animals, risk factor assessment, economic profile analysis, and georeferencing of the properties. The blood serum obtained was subjected to the AGID test. The milk samples were cultured on blood agar, and the reading was performed 24 hours after incubation. Overall, 3.4% of the animals were seropositive for CAE, and 15.8% were diagnosed with subclinical mastitis. 71.95% of the positive animals belonged to properties with a semi-intensive rearing system, weekly cleaning of the facilities, and no pre- or post-dipping. Subclinical mastitis caused by the main agent Staphylococcus spp. was associated with the breed type of the animals, highlighting two risk factors: purebred animals (OR = 12.52) and the milking performed close the soil (OR = 18.07). A reduction of up to 35.34% was observed in the income of dairy properties, constituting a considerable economic impact. Due to these sanitary problems, the economic impact demonstrates the need for a sanitary management program focused on these infirmities.

Evaluación de PCR en tiempo real en el diagnóstico de leucosis enzoótica bovina en una raza local de Panamá

El objetivo de este trabajo fue evaluar la prueba de RT-PCR, para detectar el virus de leucosis enzoótica bovina y compararlas con las pruebas de nPCR y AGID en 42 animales de la raza Guaymí de 5 regiones de Panamá. La prueba de AGID identificó el menor número de animales positivos (52%) comparada con la nPCR (71%) y PCR en tiempo real, RT-PCR (76%). Cuatro animales positivos a la prueba de AGID fueron negativos a la prueba de nPCR y dos animales positivos a esta misma prueba fueron negativos a la RT-PCR. 11 de los 20 animales negativos por AGID dieron positivo a nPCR y 12 de los 20 animales negativos a AGID fueron positivos a RT-PCR. Respecto a las pruebas moleculares, de los 12 animales que resultaron negativos a la prueba de nPCR, cinco resultaron positivos a la RT-PCR y de los 10 animales negativos a RT-PCR, 3 resultaron positivos a nPCR. La sensibilidad del nPCR respecto a AGID fue de 82% mientras que la especificidad fue de 40%. La prueba de concordancia entre pruebas fue de k= 0.316, considerada débil mientras que la sensibilidad del RT-PCR respecto a la AGID fue de 91% con una especificidad de 40%; la prueba de concordancia entre ambas pruebas fue de k=0.222 considerada también débil. La técnica de RT-PCR se presenta como una alternativa viable dentro del grupo de técnicas para el diagnóstico del virus de la leucosis enzoótica bovina, particularmente en un eventual programa de control y su erradicación en centros de conservación de razas criollas donde se requiere mantener un estricto control de la salud animal.

Rapid detection of equine infectious anaemia virus nucleic acid by insulated isothermal RT-PCR assay to aid diagnosis under field conditions.

Control of equine infectious anaemia (EIA) currently depends on serological diagnosis of infected equids. However, recently infected equids may not produce detectable anti-EIAV antibodies up to 157 days post infection and so present a high transmission risk. Therefore, direct nucleic acid detection methods are urgently needed to improve EIAV surveillance and management programs in counties where the disease is endemic. To evaluate a field-deployable, reverse transcription-insulated isothermal PCR (RT-iiPCR) assay targeting the conserved 5' untranslated region (5' UTR)/exon 1 of the tat gene of EIAV. The analytical and clinical performance of the newly developed EIAV RT-iiPCR was evaluated by comparison with a EIAV real-time RT-PCR (RT-qPCR) along with the AGID test. Analytical sensitivity was determined using in vitro transcribed RNA containing the target area of the 5' UTR/tat gene and samples from two EIAV-positive horses. Specificity was verified using nine common equine viruses. Clinical performance was evaluated by comparison with EIAV RT-qPCR and AGID using samples derived from 196 inapparent EIAV carrier horses. EIAV RT-iiPCR did not react with other commonly encountered equine viruses and had equivalent sensitivity (95% detection limit of eight genome equivalents), with a concordance of 95.41% to conventional EIAV RT-qPCR. However, the RT-qPCR and RT-iiPCR had sensitivities of 43.75 and 50.00%, respectively, when compared to the AGID test. Low viral loads commonly encountered in inapparent EIAV carriers may limit the diagnostic sensitivity of RT-PCR-based tests. Although EIAV RT-iiPCR is not sufficiently sensitive to replace the current AGID test, it can augment control efforts by identifying recently exposed or "serologically silent" equids, particularly as the latter often represent a significant transmission risk because of high viral loads. Furthermore, the relatively low cost and field-deployable design enable utilisation of EIAV RT-iiPCR even in remote regions.

Pathogenicity of Avipoxviruses in Chickens Isolated from Field Outbreaks Reported in Chhattisgarh

Virulence of field isolates of Avipoxviruses was assayed by pathogenicity test performed in 5 weeks old unvaccinated chickens. Viruses as dry scab were collected from naturally pox infected chickens, turkeys and pigeon and propagated in CAM of embryonated chickens upto various passages. In two separate trials 1 and 2, the chickens were infected with 5th and 20th passage CAM suspension, respectively by feather follicle method. All chicken groups in both trials (except control group) developed primary lesions as ‘take’ reaction from 48 to 72 hr PI and there after further progressive development of primary lesion did not differ among field isolates. In trial 1, secondary stage began with recovery from primary lesions at feather follicle, spread of infection to comb and wattles with development of secondary pox lesions and finally recovery from disease was observed after 15 days in FPV and TPV infected chickens, but not in PPV infected chickens. In trial 2, secondary pox lesions were not observed in any of the chickens, indicating that 20 passage virus induced ‘take’ at site without further spread of infection. All the recovered birds and controlled birds were challenged with the virulent FPV. The result has indicated 100% survival of birds except in control birds. Precipitating antibodies was confirmed in all birds group except control group using AGID and CIE test.

Seroepidemiological study of maedi-visna in sheep in Ceara, Rio Grande do Norte, Paraíba, and Sergipe States

The production performance of a livestock herd can be compromised by various diseases. In sheep, maedi-visna (MV) infections, which have a chronic nature, are caused by a virus (maedi-visna virus (MVV)) belonging to the genus Lentivirus of the Retroviridae family. The infection can cause significant economic losses and has considerable health impacts on sheep breeding in production systems. Due to the importance of this disease in sheep flocks, the objective was to conduct a serosurvey of MVV in the states of Ceará (CE), Rio Grande do Norte (RN), Paraíba (PB), and Sergipe (SE). A total of, 3332 serum samples were collected in the four states, 1011 in CE, 931 in RN, 459 in PB, and 931 in SE, with the number of samples proportional to the actual herd size of each state. The samples were analyzed using the agar gel microimmunodiffusion test (AGID). Reproducers were revaluated using western blotting (WB). In addition to this serological survey, we administered an investigative questionnaire to identify possible risk factors that facilitate the introduction and spread of diseases (location, category, sex, breed type, creation system, production, herd size, and association with goats). After analysis of the sera using the AGID test, there was zero prevalence. Revaluating breeders by WB revealed a 5.5% prevalence of MV in the four states studied, with prevalences for the states of CE, RN, Paraiba, and SE of 2.3% (2/88), 10.4% (8/77), 3.6% (1/28), and 4.7% (2/42), respectively, corresponding to 13 breeders containing antibodies to the virus. These findings emphasized that the choice of diagnostic tests is extremely important for the early detection of seropositive animals and thus the prevention of the spread of the virus among herds in the region.

Progressive Pathological Changes in Sub-clinical Experimental Para-tuberculosis in Goats

An experiment was carried out to study the development of sequential histological lesions and efficacy of certain diagnostic tests in experimentally induced subclinical paratuberculosis infection in goats. Twelve goats of 8-12 weeks age were infected with 4.23 x 109 Mycobacterium avium subsp. Paratuberculosis on 8 occasions. Eight goats were kept as in-contact controls and 4 as uninfected controls. Diagnostic tests, gross and histopathological studies were carried out at 3, 6, 9 and 12 months post-infection. Two goats were positive in the AGID test at 12MPI. Three goats, one at 9 MPI and two at 12 MPI were positive in faecal culture. Tissue PCR detected one goat positive each at 9 and 12 MPI. Six (50%) of 12 infected goats had gross and histological lesions. Marked enlargements of the mesenteric lymph nodes were observed in the early stages (3 and 6 months post infection) of infection. Mucosal thickening and corrugation of the jejunal and ileal mucosae, enlargement and oedema of the mesenteric lymph nodes, dilatation of lymphatics and gelatinisation of mesenteric fat were observed in goats euthanised at 9 and 12 MPI. Histological lesions consisting of flat and broad villi with mild infiltration of lymphocytes and macrophages in the intestinal villi and crypts were common at 3 MPI and 6 MPI. Focal infiltration of macrophages in Peyer's patches and giant cells were found in the mesenteric lymph nodes at 6 MPI. Broadened villi and increased infiltration of lymphocytes and macrophages with Langhan's giant cells were observed in the villi and crypts at 9 MPI and 12 MPI goats. Acid-fast bacilli were detected in 3 goats, one each at 6, 9 and 12 MPI.

Equine infectious anemia prevalence in feral donkeys from Northeast Brazil

Equine infectious anemia virus (EIAV) is an important cause of morbidity and mortality throughout the world. Although the virus infects all members of the Equidae the vast majority of studies have been conducted in horses (Equus caballus) with comparatively little information available for other equid species. Brazil has one of the most abundant donkey (E. asinus) populations of any nation although the economic importance of these animals is declining as transportation becomes increasingly mechanized. As a result, considerable numbers of donkeys especially in the Northeast of the country have been released and allowed pursue an almost feral existence. Consequently, this large and growing population constitutes a significant risk as a reservoir for the maintenance and transmission of important equine infectious diseases such as glanders and equine arteritis virus in addition to EIAV. This study examines the prevalence of EIA in a semi-wild donkey population from Mossoró city, in Northeast Brazil, using AGID followed by cELISA, rgp90 ELISA and immunoblot (IB). Serum samples were collected from 367 donkeys without obvious EIA clinical signs. Subsequent testing revealed seropositive rates of 1.6% (6/367) in officially approved AGID tests, 3.3% (12/367) in cELISA and 14.4% (53/367) in the rgp90 ELISA. However, 88.7% (47/53) of the rgp90 ELISA positive samples were almost certainly false reactions because they failed to react with two or more antigens in IB. Consequently, the rpg90 ELISA has a similar sensitivity to AGID with donkey serum samples. Such high false positive rates have not been observed previously with serum samples from horses. Another highly significant finding is that 56.9% (33/58) of the donkey serum samples tested in IB had reactivity to EIAV p26 only. Although this could result from recent infection with the virus, it has been found that in some equids p26 only reactivity persists for extensive periods of time suggesting exposure to antigens possessing cross-reactive determinants or EIAV strains with envelope glycoproteins that are different from any that have been previously characterized and so undetectable by current IB techniques.

Serologically silent, occult equine infectious anemia virus (EIAV) infections in horses

Molecular and serological techniques for Equine Infectious Anemia Virus (EIAV) diagnosis were compared using samples from 59 clinically normal horses stabled on five farms in the Santa Fe Province of Argentina. Of these 26 (44.1%) were positive in official AGID tests and/or gp45/gp90-based ELISA. Surprisingly 18 of the 33 seronegative horses were positive in a PCR against viral sequences encoding gp45 (PCR-positive/AGID-negative) with all but one remaining EIAV-antibody negative throughout a two year observation period. The gp45 PCR results are supported by fact that 7/18 of these horses were positive in the Office International des Epizooties (OIE) recommended EIAV gag gene specific PCR plus 2 of this 7 also reacted in a PCR directed predominantly against the 5′ untranslated region of the viral genome. Furthermore sufficient quantities of serum were available from 8 of these horses to verify their seronegative status in sensitive Western Blot tests and demonstrate by ELISA the absence of EIAV-specific antibodies was not attributable to abnormalities in total IgG concentration. Studies involving 7 of the PCR-positive/AGID-negative horses to measure lymphocyte proliferation in the presence of PHA showed no significant differences between this group and control animals. In addition, lymphocytes from 2 of these 7 horses responded to peptides derived from gp90 and gp45. Together these results demonstrate that apparently clinically normal horses with no gross signs of immunodeficiency in terms of total IgG concentration or T helper-cell function can remain seronegative for at least 24 months while harboring EIAV specific nucleic acid sequences.

Equine Infectious Anemia diagnosis in a 3 months old foal

within the insect, the infectious virus can remain in its mouthparts for several hours after a bite. Several studies have shown that EIAV infection has occurred among horses in North and South America, France, Germany, Italy and Romania. So far no evidence of the presence of EIAV in horses was reported in Serbia. In order to determine EIA prevalence among the 5000 horses of the Vojvodina region of Serbia, 316 horse sera have been collected during 2013 and 2014. Serological analysis was performed using Agar Gel Immuno Diffusion test (AGID) as described by the World Organization of Animal Health (OIE) chapter 2.5.6 and by Enzyme Linked Immuno Sorbent Assay (ELISA). Those sera have been tested with 3 different commercial AGID test (Idexx, ID-Vet and VMRD) and two different commercial ELISA tests (Synbiotics and VMRD). With the three AGID kit 311 (98,4%) among the 316 tested sera were found negative and only five (1,6%) sera were found positive for EIA. With ELISA kits results were slightly different and were dependent of the kit producer. Indeed, Synbiotics ELISA gave the same results as AGID tests but VMRD ELISA found 295 negative samples, 5 positive samples and 16 samples were doubtfull. Our study shows for the first time that EIA is present in Serbia and more specifically in the Vojvodina region. Euthanasia and viral characterization of field isolate collected in Serbian horses will be investigated.

Analysis of the large intestinal and faecal microbiota of horses with grass sickness using denaturing gradient gel electrophoresis

within the insect, the infectious virus can remain in its mouthparts for several hours after a bite. Several studies have shown that EIAV infection has occurred among horses in North and South America, France, Germany, Italy and Romania. So far no evidence of the presence of EIAV in horses was reported in Serbia. In order to determine EIA prevalence among the 5000 horses of the Vojvodina region of Serbia, 316 horse sera have been collected during 2013 and 2014. Serological analysis was performed using Agar Gel Immuno Diffusion test (AGID) as described by the World Organization of Animal Health (OIE) chapter 2.5.6 and by Enzyme Linked Immuno Sorbent Assay (ELISA). Those sera have been tested with 3 different commercial AGID test (Idexx, ID-Vet and VMRD) and two different commercial ELISA tests (Synbiotics and VMRD). With the three AGID kit 311 (98,4%) among the 316 tested sera were found negative and only five (1,6%) sera were found positive for EIA. With ELISA kits results were slightly different and were dependent of the kit producer. Indeed, Synbiotics ELISA gave the same results as AGID tests but VMRD ELISA found 295 negative samples, 5 positive samples and 16 samples were doubtfull. Our study shows for the first time that EIA is present in Serbia and more specifically in the Vojvodina region. Euthanasia and viral characterization of field isolate collected in Serbian horses will be investigated.

Argentinean agid test for diagnosis of equine infectious anemia: six years of history

Fil: Bok, Marina. Instituto Nacional de Tecnologia Agropecuaria. Centro de Investigacion en Ciencias Veterinarias y Agronomicas. Instituto de Virologia; Argentina. Consejo Nacional de Investigaciones Cientificas y Tecnicas; Argentina

Intercorrência entre leucose enzoótica e tuberculose em bovinos abatidos em frigorífico

RESUMO: Leucose e tuberculose são doenças que acometem o rebanho bovino e acarretam consideráveis prejuízos econômicos, em virtude do impacto na produtividade, porém há poucos relatos de que elas ocorram concomitantemente. Diante disso, objetivou-se avaliar a intercorrência entre essas doenças em bovinos abatidos no município de São Gotardo, MG, sendo que esses animais foram reagentes à tuberculinização e apresentaram lesões características da doença durante o abate sanitário. Cento e trinta e três amostras de soro sanguíneo, todas oriundas de bovinos positivos para tuberculose, foram analisadas pela técnica de imunodifusão em gel de ágar (IDGA), e a prevalência de bovinos que apresentaram positividade ao teste de IDGA foi de 55,64% (74/133). Os resultados obtidos nesta pesquisa permitiram concluir que a tuberculose e a leucose são doenças presentes nas regiões estudadas e encontram-se disseminadas na população avaliada, com níveis significativos, causando risco à saúde dos animais e das pessoas, dado o caráter zoonótico da tuberculose bovina.

Characterization of a new influenza virus type D

Influenza is caused by viruses belonging to the Orthomyxoviridae family. Currently three types of influenza virus are known: A (Influenza A virus, IAV), B (IBV) and C (ICV). Despite the fact that all these viruses are derived from a common ancestor they differ from each other by the number of segments, the size and sequence of RNA segments, antigenicity, pathogenicity and the spectrum of natural reservoirs. In 2011, a new influenza virus was isolated in the USA from pigs manifesting influenza-like symptoms. The virus was the most closely related to ICV. It was able to replicate in vitro in different cell cultures and displayed much broader cell tropism than human ICV. Moreover, in contrast to ICV, it was able to replicate at 370C. Electron microscopic studies demonstrated features characteristic of Orthomyxoviruses. Despite morphological and organizational similarities, the biological properties of the new virus, including biochemical activity, differ from that of other influenza viruses. Enzymatic assays revealed that the new virus had negligible neuraminidase but detectable O-acetyloesterase activity. Further studies evidenced that the new virus varied from ICV in receptor binding, despite its sharing a conserved array of functional domains in the viral RNA genome replication and viral entry machinery. Analysis conducted with the use of the model of crystal structure of the hemagglutinin-esterase fusion protein (HE) of the new virus and its receptor demonstrated that this protein was multifunctional. It catalyzes cellular receptor binding, receptor cleavage, as well as membrane fusion. Moreover, divergent receptor-binding sites than HE of ICV have been discovered in the new virus. These amino acid differences may alter the binding specificity and affinity of the HE protein to the receptor that in turn result in the observed differences in cellular tropism between the two viruses. It also possesses an open channel between the 230-helix and 270-loop in the receptor-binding site, which is a unique feature of this virus. This might explain why the new virus has a broad cell tropism. It is possible that the sequence variation in the fusion domain may influence the replication of this virus at a higher temperature when compared to ICV. Next-generation sequencing demonstrated that the genome of the new virus, similarly to ICV, had seven single-stranded negative-sense RNA segments coding 9 viral proteins. Deep RNA sequencing found a M1 protein expression strategy different from that of ICV. Studies aimed at evaluating of the evolutionary relationship of both viruses revealed that the new virus and ICV shared an approximately 69-72% mean pairwise identity in the PB1 gene, which is reported to be the most conserved influenza virus protein. Additionally, differences were detected at 5’ and 3’ends of noncoding regions, which are also highly conserved. They both may be responsible for the lack of in vitro reassortment between ICV of human origin and the new virus. In the study characterizing antigenic properties of the new virus, no cross-reactivity was observed using HI and AGID tests. This indicates the major differences in conserved proteins M1 and NP between both viruses. Summing up, despite the fact that new virus is the most closely related to human ICV, the number of important antigenic and genetic distinctions among them is the basis for suggesting that the International Committee of Virus Taxonomy classify it as a separate genus - D. There is no doubt that the discovery of a new influenza virus genus will have a great impact on influenza research and ecology.

SEROSURVEILLANCE OF EQUINE INFECTIOUS ANAEMIA IN A REGION OF VOJVODINA

Equine infectious anemia is a consequence of a persistant infection of the horses with Lentivirus. Pathogenesis of the disease is very variable, what can bee seen through a wide range of clinical forms of the disease – from inaparrent infection to death. Diagnostics of EIA is based on clinical symptoms, detection of antibodies and virus. Antibodies can be identified with Hi, VN, CFIT, cELISA, SA-ELISA and AGID test. RT-PCR technique enables the detection of and/or quantifi cation of viral RNA level in blood of infected animal. First reliable serological test for EIA was AGID test. Modified AGID test is considered today as aknowledged, international standard for the detection of antibodies against EIA virus and it enables detection of more then 95% of ll positive animals. Horses with positive fi ndings with this test are considered infected and should be euthanized or placed in strict isolation. Further measures to control the spread of this disease are insectvector control and disinfection of surgical and other equipment in use on successive animals. Th e results of a study during a twenty year period, in the region of AP Vojvodina show that from the total of 11.972 horses blood samples, with te use of AGID test, positive results were found in 21 or 0,17% of horses.

Development of an AGID based on baculovirus expressed Pseudorabies virus glycoprotein B

Development of an AGID based on baculovirus expressed Pseudorabies virus glycoprotein B

The immunological & serological studies of Equine Infectious Anemia in Egyptian horses

Equine infection anemia (EIA) occurs world-wide. The infection, formerly known as swamp fever, is limited to equids.The disease is characterized by recurrent febrile episodes, thrombocytopenia and anemia We collected 10 cc. of aseptically collected whole blood samples from 180 horses at different localities in Egypt suffering from fever, illness, anemia and infertility. Samples may be submitted as serum or whole blood should be collected in vacutainers without anti-coagulant using disposable needles and sterile syringes. Samples must be properly identified, carefully packed, and sent as soon as possible to diagnostic lab. Clinical diagnosiswas done by applied Agar gel immunodiffusion and ELISA tests. In this study recorded that 33 (18.33%) positive EIA by using ELISA test but was 24 (13.33%) by AGID test. The highest incidence was in Kaliobia Gov.8/35 samples 22.85% followed by Cairo 9/50 samples 18%, then Giza 11/65 samples 16.92% and Fayoum 5/30 samples 16.66% by ELISA test. The same results were inKaliobia Gov. 5/35 samples 14.29% followed by Cairo 7/50 samples 14% then 9/65 samples 13.85% in Giza and 3/30 samples 10% in Fayoum by using AGID test. Finally, we concluded that combining the specificity of AGID test with higher sensitivity, such as the ELISA test to enhance the accuracy of EIA diagnosis.

Production of equine herpesvirus 1 recombinant glycoprotein D and development of an agar gel immunodiffusion test for serological diagnosis

Equine herpesvirus 1 and 4 (EHV-1 and 4) infect most of the world's horses, causing serious clinical illness. Viral glycoproteins have been identified as the immunodominant antigens that generate the antiviral serological responses to EHV-1 and EHV-4 in infected horses. Here, glycoprotein D of EHV-1 was expressed by a recombinant baculovirus, purified and evaluated by a simple agar gel immunodiffusion test (AGID). Compared with virus neutralization, serological analysis by AGID showed good specificity (100%) and sensitivity (99.5%). The estimated Kappa values for repeatability and reproducibility were satisfactory. Thus, this rapid, inexpensive, simple and highly specific AGID test seems to be a valuable alternative tool for serological detection of antibodies against both EHV-1 and EHV-4.

Molecular epidemiology of Equine Arteritis Virus in France following the 2007 outbreak

clinical signs. Five were from France and five came from Romania. Positive horses identified in 2010 were from 7 different premises divided in two different episodes. The first episode detected in South West of France consisted of 4 related premises where 5 French Trotters were tested positives and euthanized. The index case has been tested as part of a control before exportation. Primary premise located in the Dordogne County was a breeding farm where 2 mares were tested positive for EIA. Others premises, where two positive horses were detected, were located in the Lot-et-Garonne and Gironde County. All Infected animals (5) came from the breeding farm in Dordogne and have been kept over there several years where they have been probably infected since it is the only source of infection identified. Epidemiological investigations lead to test more than 400 horses, in 38 different Counties, which have been trained in the Dordogne Farm between 1990 and 2010. No other positive animals have been identified during this seroepidemiological study. The second EIA episode identified in 2010 was from horses which have been imported from Romania. Indeed, following EIA cases identified in Belgium and in UK at the end of 2009, French Ministry of Agriculture decided to test horses that came from Romania since January 2007. Investigation detected 80 equids, among those horses 38 were slaughtered or re-exported. Only 35 horses were still on the French territory at the date of testing and five of them exhibited a positive result for EIA using AGID test. Horses kept at the vicinity of those positive horses were tested but no transmission was seen. In this context the European commission (EU) has modified in June 2010 the regulation regarding importation of horses from Romania in other European member states. EU enforced a quarantine and a testing at destination for horses coming from Romania.