Aino Virus

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Tomoyuki Tsuda - One of the best experts on this subject based on the ideXlab platform.

  • molecular epidemiological analyses of the teratogenic Aino Virus based on the sequences of a small rna segment
    Veterinary Microbiology, 2008
    Co-Authors: Makoto Yamakawa, Tohru Yanase, Tomoko Kato, Tomoyuki Tsuda
    Abstract:

    The sequences of a small RNA segment of Aino Virus isolates were analyzed to define the molecular epidemiology and genetic relationships to other species in the genus OrthobunyaVirus in the family Bunyaviridae. The nucleotide and amino acid sequences of the segment were highly conserved among strains isolated from 1964 to 2002 in Japan. These Japanese isolates were segregated into two distinct lineages, one containing the prototype strain JaNAr28 isolated in 1964 and the other containing strains isolated after 1986, by phylogenetic analysis based on the nucleocapsid gene sequences. Japanese strains isolated after 1986 were rather more closely related to Kaikalur Virus isolated in India in 1971 than to strain JaNAr28. On the other hand, an Australian strain, B7974, was closely related to Peaton Virus. The B7974 strain might have been generated by inter-serotype genetic reassortment between Aino and Peaton Viruses in Australia during their evolution. However, recent Aino Virus strains isolated in Japan appear to be genetically stable.

  • arthrogryposis hydranencephaly and cerebellar hypoplasia syndrome in neonatal calves resulting from intrauterine infection with Aino Virus
    Veterinary Research, 2004
    Co-Authors: Tomoyuki Tsuda, Kazuo Yoshida, Seiichi Ohashi, Tohru Yanase, Masuo Sueyoshi, Syunichi Kamimura, Kazuhiro Misumi, Katsumi Hamana, Hiroshi Sakamoto, Makoto Yamakawa
    Abstract:

    To determine the teratogenic potential of Aino Virus (AinoV) in cattle, pregnant cows and fetal cattle were infected with a fresh isolate of AinoV. Five pregnant cows were inoculated intravenously with the Virus at 122 to 162 days of gestation and allowed to give birth. All of the cows developed neutralizing antibodies to the Virus, indicating that the cows had been infected with the Virus; however, no clinical abnormalities were seen in their six newborn calves, and no specific antibodies to the Virus were detected in the precolostral serum of calves. Five fetuses with fetal ages ranging from 132 to 156 days were inoculated in utero with the Virus. One weak newborn and four stillborn calves were delivered at gestation days 256 to 263, i.e., less than the standard gestation term; they had congenital abnormalities including arthrogryposis, hydranencephaly and cerebellar hypoplasia. Antibodies specific to AinoV were detected in their precolostral serum. These results demonstrate that AinoV is a potential etiological agent of congenital malformation of cattle.

  • Simultaneous detection of bovine arboViruses using single-tube multiplex reverse transcription-polymerase chain reaction
    Journal of Virological Methods, 2004
    Co-Authors: Seiichi Ohashi, Kazuo Yoshida, Tohru Yanase, Tomoko Kato, Tomoyuki Tsuda
    Abstract:

    Abstract Single-tube multiplex reverse transcription-polymerase chain reaction (mRT-PCR) assay was developed to detect and identify arboViruses in infected cell-culture fluids and field specimens. The technique was equally sensitive for detecting five different Viruses in cell cultures, namely the Chuzan, Ibaraki, and Bluetongue Viruses belonging to OrbiVirus, and the Akabane Virus and Peaton Virus belonging to OrthobunyaVirus, and was less sensitive than former Viruses for detecting Aino Virus belonging to OrthobunyaVirus. The mRT-PCR reliably detected 0.6–103.1 median tissue culture infective doses. The mRT-PCR readily identified Viruses by discriminating the size of their amplified gene products. The technique was as sensitive as Virus isolation in detecting single infected plasma in five plasmas from sentinel cattle and in detecting two infectious homogenates in eight homogenates of Culicoides biting midges. The mRT-PCR may be a sensitive and rapid assay for surveillance of bovine arboViruses in field specimens.

  • Serological and clinical evidence of a teratogenic Simbu serogroup Virus infection of cattle in Israel, 2001-2003.
    Veterinaria Italiana, 2004
    Co-Authors: J. Brenner, Tomoyuki Tsuda, Yehuda Stram, Dalia Chai, Hagai Yadin, Tomoko Kato
    Abstract:

    During the last 35 years, two major outbreaks of Akabane Virus (AKAV) infection were recorded in cattle in Israel in 1969/1970 and 2002/2003. Congenital malformations of calves characterised by the appearance of an arthrogryposis and hydranencephaly syndrome first appeared in Israel in 1969. Based on epidemiological, clinical, pathological, histopathological and serological data, this syndrome was strongly correlated with seroreactivity to AKAV, a member of the Bunyaviridae, Simbu serogroup. In February 2002, the first cases of 'blind newborn calves' (BNC) were observed on farms located in the northern valleys of Israel. Microtitre serum neutralisation (SN) tests of serum from malformed calves and their dams were conducted using Akabane and Aino Viruses (AinoV). The first SN test was performed at the reference laboratory of the Clinical Virology Section, Kyushu Research Station, National Institute of Animal Health, Kagoshima, Japan. The clear-cut findings of seroreactivity to AKAV by cattle located in the affected zone, in contrast to negative findings in cattle from unaffected farms (87% and 3.7%, respectively) was indicative of AKAV infection. In contrast, seroreactivity to Aino Virus was relatively low in both affected and non-affected areas during the 2002 outbreak. In order to establish Israeli laboratory standards for Simbu serogroup diagnosis, 57 serum samples tested by the Japanese laboratory were retested by SN in Israel. An almost complete homology (96.5%) was found between the two SN panels of sera (kappa = 0.92). SN and ELISA kits enabled the surveillance of this arboVirus epidemic in the second consecutive year (2003). Moreover, AKAV was identified in trapped midges by hemi-nested PCR and real-time PCR. With these techniques, the geographical limits of the BNC epidemic that appeared in some areas of Israel was identified for the first time and was recorded in the Arava Rift Valley, 400 km south of the epicentre of the 2002 outbreak. The reintroduction of AKAV into this region, together with some evidence of AinoV activity and epidemics of bluetongue (BT) in the southern parts of Europe and the eastern Mediterranean, and renewed outbreaks of West Nile Virus infection in Israel, Italy and southern France, are all evidence of the potential spread of arboVirus activity into southern Europe from the Mediterranean Basin.

  • Sequence analysis of the medium RNA segment of three Simbu serogroup Viruses, Akabane, Aino, and Peaton Viruses ☆
    Virus Research, 2003
    Co-Authors: Tohru Yanase, Kazuo Yoshida, Seiichi Ohashi, Tomoko Kato, Tomoyuki Tsuda
    Abstract:

    The sequence analysis was carried out for the medium (M) RNA segment of the Akabane Virus (AKAV), Aino Virus (AINV), and Peaton Virus (PEAV) of the Simbu serogroup of the genus OrthobunyaVirus of the family Bunyaviridae. The complementary sequences of the M RNA segments of AKAV, AINV, and PEAV contain a single large open reading frame (ORF), like other orthobunyaViruses. The ORFs potentially encode 1401 amino acids (aa), 1404 aa, and 1400 aa polypeptides, respectively. The identity of the M segment among these Viruses is remarkably low, although previous researchers reported that the small RNA segments are highly conserved. Because the M segment codes for the viral surface glycoproteins G1 and G2, the variability of the M segment may affect the antigenicity of these Viruses. Phylogenetic studies based on the M and S segment sequences suggested that genetic reassortment has been occurring among ancestral Viruses of the three Simbu serogroup Viruses throughout their evolution.

Kazuo Yoshida - One of the best experts on this subject based on the ideXlab platform.

  • Simultaneous detection of bovine arboViruses using single-tube multiplex reverse transcription-polymerase chain reaction
    Journal of Virological Methods, 2004
    Co-Authors: Seiichi Ohashi, Kazuo Yoshida, Tohru Yanase, Tomoko Kato, Tomoyuki Tsuda
    Abstract:

    Abstract Single-tube multiplex reverse transcription-polymerase chain reaction (mRT-PCR) assay was developed to detect and identify arboViruses in infected cell-culture fluids and field specimens. The technique was equally sensitive for detecting five different Viruses in cell cultures, namely the Chuzan, Ibaraki, and Bluetongue Viruses belonging to OrbiVirus, and the Akabane Virus and Peaton Virus belonging to OrthobunyaVirus, and was less sensitive than former Viruses for detecting Aino Virus belonging to OrthobunyaVirus. The mRT-PCR reliably detected 0.6–103.1 median tissue culture infective doses. The mRT-PCR readily identified Viruses by discriminating the size of their amplified gene products. The technique was as sensitive as Virus isolation in detecting single infected plasma in five plasmas from sentinel cattle and in detecting two infectious homogenates in eight homogenates of Culicoides biting midges. The mRT-PCR may be a sensitive and rapid assay for surveillance of bovine arboViruses in field specimens.

  • arthrogryposis hydranencephaly and cerebellar hypoplasia syndrome in neonatal calves resulting from intrauterine infection with Aino Virus
    Veterinary Research, 2004
    Co-Authors: Tomoyuki Tsuda, Kazuo Yoshida, Seiichi Ohashi, Tohru Yanase, Masuo Sueyoshi, Syunichi Kamimura, Kazuhiro Misumi, Katsumi Hamana, Hiroshi Sakamoto, Makoto Yamakawa
    Abstract:

    To determine the teratogenic potential of Aino Virus (AinoV) in cattle, pregnant cows and fetal cattle were infected with a fresh isolate of AinoV. Five pregnant cows were inoculated intravenously with the Virus at 122 to 162 days of gestation and allowed to give birth. All of the cows developed neutralizing antibodies to the Virus, indicating that the cows had been infected with the Virus; however, no clinical abnormalities were seen in their six newborn calves, and no specific antibodies to the Virus were detected in the precolostral serum of calves. Five fetuses with fetal ages ranging from 132 to 156 days were inoculated in utero with the Virus. One weak newborn and four stillborn calves were delivered at gestation days 256 to 263, i.e., less than the standard gestation term; they had congenital abnormalities including arthrogryposis, hydranencephaly and cerebellar hypoplasia. Antibodies specific to AinoV were detected in their precolostral serum. These results demonstrate that AinoV is a potential etiological agent of congenital malformation of cattle.

  • Sequence analysis of the medium RNA segment of three Simbu serogroup Viruses, Akabane, Aino, and Peaton Viruses ☆
    Virus Research, 2003
    Co-Authors: Tohru Yanase, Kazuo Yoshida, Seiichi Ohashi, Tomoko Kato, Tomoyuki Tsuda
    Abstract:

    The sequence analysis was carried out for the medium (M) RNA segment of the Akabane Virus (AKAV), Aino Virus (AINV), and Peaton Virus (PEAV) of the Simbu serogroup of the genus OrthobunyaVirus of the family Bunyaviridae. The complementary sequences of the M RNA segments of AKAV, AINV, and PEAV contain a single large open reading frame (ORF), like other orthobunyaViruses. The ORFs potentially encode 1401 amino acids (aa), 1404 aa, and 1400 aa polypeptides, respectively. The identity of the M segment among these Viruses is remarkably low, although previous researchers reported that the small RNA segments are highly conserved. Because the M segment codes for the viral surface glycoproteins G1 and G2, the variability of the M segment may affect the antigenicity of these Viruses. Phylogenetic studies based on the M and S segment sequences suggested that genetic reassortment has been occurring among ancestral Viruses of the three Simbu serogroup Viruses throughout their evolution.

  • comparison of intertypic antigenicity of Aino Virus isolates by dot immunobinding assay using neutralizing monoclonal antibodies
    Journal of Clinical Microbiology, 2000
    Co-Authors: Kazuo Yoshida, Seiichi Ohashi, Tomomi Kubo, Tomoyuki Tsuda
    Abstract:

    Neutralizing monoclonal antibodies (MAbs) against the Aino Virus were prepared, and the neutralizing epitopes of the Virus were defined by competitive binding assay. Seven continuous and overlapping neutralizing epitopes existed on the G1 glycoprotein of the Aino Virus. Two antigenic domains were identified and were designated I and II, with domain II consisting of six epitopes. Dot immunobinding assays (DIAs) were performed with MAbs that recognized these seven neutralizing epitopes. DIAs were performed with 1 Australian strain and 21 isolates found in Japan between the years 1964 and 1995. The MAb response patterns of all isolates were divided into four groups. The Japanese isolates did not show large differences in antigenicity, but the antigenicity of the Australian strain collected in 1968 was significantly different from that of the Japanese strains; the Australian strain lacked reactivity to three epitopes and showed only low reactivity to one epitope.

Tohru Yanase - One of the best experts on this subject based on the ideXlab platform.

  • molecular epidemiological analyses of the teratogenic Aino Virus based on the sequences of a small rna segment
    Veterinary Microbiology, 2008
    Co-Authors: Makoto Yamakawa, Tohru Yanase, Tomoko Kato, Tomoyuki Tsuda
    Abstract:

    The sequences of a small RNA segment of Aino Virus isolates were analyzed to define the molecular epidemiology and genetic relationships to other species in the genus OrthobunyaVirus in the family Bunyaviridae. The nucleotide and amino acid sequences of the segment were highly conserved among strains isolated from 1964 to 2002 in Japan. These Japanese isolates were segregated into two distinct lineages, one containing the prototype strain JaNAr28 isolated in 1964 and the other containing strains isolated after 1986, by phylogenetic analysis based on the nucleocapsid gene sequences. Japanese strains isolated after 1986 were rather more closely related to Kaikalur Virus isolated in India in 1971 than to strain JaNAr28. On the other hand, an Australian strain, B7974, was closely related to Peaton Virus. The B7974 strain might have been generated by inter-serotype genetic reassortment between Aino and Peaton Viruses in Australia during their evolution. However, recent Aino Virus strains isolated in Japan appear to be genetically stable.

  • arthrogryposis hydranencephaly and cerebellar hypoplasia syndrome in neonatal calves resulting from intrauterine infection with Aino Virus
    Veterinary Research, 2004
    Co-Authors: Tomoyuki Tsuda, Kazuo Yoshida, Seiichi Ohashi, Tohru Yanase, Masuo Sueyoshi, Syunichi Kamimura, Kazuhiro Misumi, Katsumi Hamana, Hiroshi Sakamoto, Makoto Yamakawa
    Abstract:

    To determine the teratogenic potential of Aino Virus (AinoV) in cattle, pregnant cows and fetal cattle were infected with a fresh isolate of AinoV. Five pregnant cows were inoculated intravenously with the Virus at 122 to 162 days of gestation and allowed to give birth. All of the cows developed neutralizing antibodies to the Virus, indicating that the cows had been infected with the Virus; however, no clinical abnormalities were seen in their six newborn calves, and no specific antibodies to the Virus were detected in the precolostral serum of calves. Five fetuses with fetal ages ranging from 132 to 156 days were inoculated in utero with the Virus. One weak newborn and four stillborn calves were delivered at gestation days 256 to 263, i.e., less than the standard gestation term; they had congenital abnormalities including arthrogryposis, hydranencephaly and cerebellar hypoplasia. Antibodies specific to AinoV were detected in their precolostral serum. These results demonstrate that AinoV is a potential etiological agent of congenital malformation of cattle.

  • Simultaneous detection of bovine arboViruses using single-tube multiplex reverse transcription-polymerase chain reaction
    Journal of Virological Methods, 2004
    Co-Authors: Seiichi Ohashi, Kazuo Yoshida, Tohru Yanase, Tomoko Kato, Tomoyuki Tsuda
    Abstract:

    Abstract Single-tube multiplex reverse transcription-polymerase chain reaction (mRT-PCR) assay was developed to detect and identify arboViruses in infected cell-culture fluids and field specimens. The technique was equally sensitive for detecting five different Viruses in cell cultures, namely the Chuzan, Ibaraki, and Bluetongue Viruses belonging to OrbiVirus, and the Akabane Virus and Peaton Virus belonging to OrthobunyaVirus, and was less sensitive than former Viruses for detecting Aino Virus belonging to OrthobunyaVirus. The mRT-PCR reliably detected 0.6–103.1 median tissue culture infective doses. The mRT-PCR readily identified Viruses by discriminating the size of their amplified gene products. The technique was as sensitive as Virus isolation in detecting single infected plasma in five plasmas from sentinel cattle and in detecting two infectious homogenates in eight homogenates of Culicoides biting midges. The mRT-PCR may be a sensitive and rapid assay for surveillance of bovine arboViruses in field specimens.

  • Sequence analysis of the medium RNA segment of three Simbu serogroup Viruses, Akabane, Aino, and Peaton Viruses ☆
    Virus Research, 2003
    Co-Authors: Tohru Yanase, Kazuo Yoshida, Seiichi Ohashi, Tomoko Kato, Tomoyuki Tsuda
    Abstract:

    The sequence analysis was carried out for the medium (M) RNA segment of the Akabane Virus (AKAV), Aino Virus (AINV), and Peaton Virus (PEAV) of the Simbu serogroup of the genus OrthobunyaVirus of the family Bunyaviridae. The complementary sequences of the M RNA segments of AKAV, AINV, and PEAV contain a single large open reading frame (ORF), like other orthobunyaViruses. The ORFs potentially encode 1401 amino acids (aa), 1404 aa, and 1400 aa polypeptides, respectively. The identity of the M segment among these Viruses is remarkably low, although previous researchers reported that the small RNA segments are highly conserved. Because the M segment codes for the viral surface glycoproteins G1 and G2, the variability of the M segment may affect the antigenicity of these Viruses. Phylogenetic studies based on the M and S segment sequences suggested that genetic reassortment has been occurring among ancestral Viruses of the three Simbu serogroup Viruses throughout their evolution.

Seiichi Ohashi - One of the best experts on this subject based on the ideXlab platform.

  • Simultaneous detection of bovine arboViruses using single-tube multiplex reverse transcription-polymerase chain reaction
    Journal of Virological Methods, 2004
    Co-Authors: Seiichi Ohashi, Kazuo Yoshida, Tohru Yanase, Tomoko Kato, Tomoyuki Tsuda
    Abstract:

    Abstract Single-tube multiplex reverse transcription-polymerase chain reaction (mRT-PCR) assay was developed to detect and identify arboViruses in infected cell-culture fluids and field specimens. The technique was equally sensitive for detecting five different Viruses in cell cultures, namely the Chuzan, Ibaraki, and Bluetongue Viruses belonging to OrbiVirus, and the Akabane Virus and Peaton Virus belonging to OrthobunyaVirus, and was less sensitive than former Viruses for detecting Aino Virus belonging to OrthobunyaVirus. The mRT-PCR reliably detected 0.6–103.1 median tissue culture infective doses. The mRT-PCR readily identified Viruses by discriminating the size of their amplified gene products. The technique was as sensitive as Virus isolation in detecting single infected plasma in five plasmas from sentinel cattle and in detecting two infectious homogenates in eight homogenates of Culicoides biting midges. The mRT-PCR may be a sensitive and rapid assay for surveillance of bovine arboViruses in field specimens.

  • arthrogryposis hydranencephaly and cerebellar hypoplasia syndrome in neonatal calves resulting from intrauterine infection with Aino Virus
    Veterinary Research, 2004
    Co-Authors: Tomoyuki Tsuda, Kazuo Yoshida, Seiichi Ohashi, Tohru Yanase, Masuo Sueyoshi, Syunichi Kamimura, Kazuhiro Misumi, Katsumi Hamana, Hiroshi Sakamoto, Makoto Yamakawa
    Abstract:

    To determine the teratogenic potential of Aino Virus (AinoV) in cattle, pregnant cows and fetal cattle were infected with a fresh isolate of AinoV. Five pregnant cows were inoculated intravenously with the Virus at 122 to 162 days of gestation and allowed to give birth. All of the cows developed neutralizing antibodies to the Virus, indicating that the cows had been infected with the Virus; however, no clinical abnormalities were seen in their six newborn calves, and no specific antibodies to the Virus were detected in the precolostral serum of calves. Five fetuses with fetal ages ranging from 132 to 156 days were inoculated in utero with the Virus. One weak newborn and four stillborn calves were delivered at gestation days 256 to 263, i.e., less than the standard gestation term; they had congenital abnormalities including arthrogryposis, hydranencephaly and cerebellar hypoplasia. Antibodies specific to AinoV were detected in their precolostral serum. These results demonstrate that AinoV is a potential etiological agent of congenital malformation of cattle.

  • Sequence analysis of the medium RNA segment of three Simbu serogroup Viruses, Akabane, Aino, and Peaton Viruses ☆
    Virus Research, 2003
    Co-Authors: Tohru Yanase, Kazuo Yoshida, Seiichi Ohashi, Tomoko Kato, Tomoyuki Tsuda
    Abstract:

    The sequence analysis was carried out for the medium (M) RNA segment of the Akabane Virus (AKAV), Aino Virus (AINV), and Peaton Virus (PEAV) of the Simbu serogroup of the genus OrthobunyaVirus of the family Bunyaviridae. The complementary sequences of the M RNA segments of AKAV, AINV, and PEAV contain a single large open reading frame (ORF), like other orthobunyaViruses. The ORFs potentially encode 1401 amino acids (aa), 1404 aa, and 1400 aa polypeptides, respectively. The identity of the M segment among these Viruses is remarkably low, although previous researchers reported that the small RNA segments are highly conserved. Because the M segment codes for the viral surface glycoproteins G1 and G2, the variability of the M segment may affect the antigenicity of these Viruses. Phylogenetic studies based on the M and S segment sequences suggested that genetic reassortment has been occurring among ancestral Viruses of the three Simbu serogroup Viruses throughout their evolution.

  • comparison of intertypic antigenicity of Aino Virus isolates by dot immunobinding assay using neutralizing monoclonal antibodies
    Journal of Clinical Microbiology, 2000
    Co-Authors: Kazuo Yoshida, Seiichi Ohashi, Tomomi Kubo, Tomoyuki Tsuda
    Abstract:

    Neutralizing monoclonal antibodies (MAbs) against the Aino Virus were prepared, and the neutralizing epitopes of the Virus were defined by competitive binding assay. Seven continuous and overlapping neutralizing epitopes existed on the G1 glycoprotein of the Aino Virus. Two antigenic domains were identified and were designated I and II, with domain II consisting of six epitopes. Dot immunobinding assays (DIAs) were performed with MAbs that recognized these seven neutralizing epitopes. DIAs were performed with 1 Australian strain and 21 isolates found in Japan between the years 1964 and 1995. The MAb response patterns of all isolates were divided into four groups. The Japanese isolates did not show large differences in antigenicity, but the antigenicity of the Australian strain collected in 1968 was significantly different from that of the Japanese strains; the Australian strain lacked reactivity to three epitopes and showed only low reactivity to one epitope.

Hye-young Jeoung - One of the best experts on this subject based on the ideXlab platform.

  • Detection and differentiation of Schmallenberg, Akabane and Aino Viruses by one-step multiplex reverse-transcriptase quantitative PCR assay
    BMC Veterinary Research, 2015
    Co-Authors: Jee-yong Park, Hye-young Jeoung
    Abstract:

    Background Schmallenberg Virus (SBV), Akabane Virus (AKAV) and Aino Virus (AINV) are members of the Simbu serogroup within the genus OrthobunyaVirus, family Bunyaviridae, which can cause reproductive disorders including abortion, stillbirth and congenital malformation in ruminants. Because, the clinical signs are similar, confirmatory diagnosis requires viral detection to differentiate infection between these three Viruses.

  • Detection and differentiation of Schmallenberg, Akabane and Aino Viruses by one-step multiplex reverse-transcriptase quantitative PCR assay
    BMC Veterinary Research, 2015
    Co-Authors: Jee-yong Park, Hye-young Jeoung
    Abstract:

    Background Schmallenberg Virus (SBV), Akabane Virus (AKAV) and Aino Virus (AINV) are members of the Simbu serogroup within the genus OrthobunyaVirus , family Bunyaviridae, which can cause reproductive disorders including abortion, stillbirth and congenital malformation in ruminants. Because, the clinical signs are similar, confirmatory diagnosis requires viral detection to differentiate infection between these three Viruses. Methods In this study, a one-step multiplex reverse-transcriptase quantitative PCR (one-step mRT-qPCR) was developed for the simultaneous detection and differentiation of SBV, AKAV and AINV. Results The detection limit of the one-step mRT-qPCR for SBV, AKAV and AINV were 2.4 copies (10 ^0.6 TCID _50/ml), 96.2 copies (10 ^1.5 TCID _50/ml) and 52.3 copies (10 ^1.2 TCID _50/ml), respectively. Various field samples such as bovine serum, bovine whole blood, bovine brain, goat serum and Culicoides were analyzed using the one-step mRT-qPCR and compared with previously published RT-qPCRs. The test results of the field samples were identical for the one-step mRT-qPCR and RT-qPCRs, which showed all samples to be negative for SBV, AKAV and AINV, except for one bovine brain sample (1/123) that was positive for AKAV. Conclusion The one-step mRT-qPCR allows for the simultaneous detection of three viral pathogens (SBV, AKAV and AINV) that cause reproductive failure.