Tohru Yanase

Genetic reassortment between Sathuperi and Shamonda viruses of the genus Orthobunyavirus in nature: implications for their genetic relationship to Schmallenberg virus

The recent outbreak of malformations in ruminants in Northern Europe caused by Schmallenberg virus induced us to analyze the genetic properties of the related orthobunyaviruses and clarify their relationship. The sequencing of three genomic RNA segments of Sathuperi, Shamonda and Douglas viruses (SATV, SHAV and DOUV) revealed that the M RNA segment of SATV and DOUV had a high degree of sequence identity with that of Schmallenberg virus, but the S and L RNA segments closely matched those of SHAV. Phylogenetic analysis of the three genomic RNA segments indicated that Schmallenberg virus is a reassortant, with the M RNA segment from SATV and the S and L RNA segments from SHAV.

Bovine epizootic encephalomyelitis caused by Akabane virus in southern Japan

BackgroundAkabane virus is a member of the genus Orthobunyavirus in the family Bunyaviridae. It is transmitted by hematophagous arthropod vectors such as Culicoides biting midges and is widely distributed in temperate to tropical regions of the world. The virus is well known as a teratogenic pathogen which causes abortions, stillbirths, premature births and congenital abnormalities with arthrogryposis-hydranencephaly syndrome in cattle, sheep and goats. On the other hand, it is reported that the virus rarely induces encephalomyelitis in cattle by postnatal infection. A first large-scale epidemic of Akabane viral encephalomyelitis in cattle occurred in the southern part of Japan from summer to autumn in 2006. The aim of this study is to define the epidemiological, pathological and virological properties of the disease.ResultsNonsuppurative encephalomyelitis was observed in cattle that showed neurological symptoms such as astasia, ataxia, opisthotonus and hypersensitivity in beef and dairy farms by histopathological analysis. Akabane viral antigen and genome were consistently detected from the central nervous system of these animals, and the virus was isolated not only from them but also from the blood samples of clinically healthy calves in the epidemic area. The isolates were classified into genogroup I a containing the Iriki strain, which caused encephalitis of calves almost twenty years ago in Japan. Most of the affected cattle possessed the neutralizing antibody against Akabane virus. Seroconversion of the cohabitated and sentinel cattle in the epidemic area was also confirmed during an outbreak of the disease.ConclusionThe ecological and epidemiological data we have obtained so far demonstrated that the Akabane virus is not endemic in Japan. No evidence of Akabane virus circulation was observed in 2005 through nation-wide serological surveillance, suggesting that a new strain belonging to genogroup I a invaded southern Japan from overseas in the summer of 2006 and caused an unprecedented epizootic of encephalomyelitis mainly in susceptible calves. It will be necessary to reconsider the vaccine strategy to control the disease effectually.

Isolation of Bovine Arboviruses from Culicoides Biting Midges (Diptera: Ceratopogonidae) in Southern Japan: 1985–2002

Abstract In 1985–2002, surveillance for bovine arboviruses was conducted in Kagoshima, located in the most southern part of the main islands of Japan and known to be an area where bovine arboviral diseases have frequently been epidemic. Culicoides biting midges were collected in a cowshed by light traps. A total of 456,300 Culicoides biting midges representing 13 species were collected, and a portion of each pool of midges were tested for virus isolation. Overall, 85 isolates of six different viruses were obtained from the collected midges. The isolated viruses included two Orthobunyaviruses, Akabane and Aino viruses; three Orbiviruses, Chuzan, D’Aguliar, and Ibaraki viruses; and one unclassified virus, a bunyavirus-like virus. The viruses were most frequently isolated from Culicoides oxystoma Kieffer (85.9% of 85 isolates). Isolations of all viruses except for the bunyavirus-like virus were made from this species. Our data indicated that C. oxystoma is a potential vector for bovine arboviruses in southern Japan.

Genetic characterization of Batai virus indicates a genomic reassortment between orthobunyaviruses in nature

Summary.Two viruses were isolated from bovine blood in the southernmost part of Japan in 1994 and 2001, respectively. Genetic analyses showed that the viruses were Batai virus of the genus Orthobunyavirus of the family Bunyaviridae. The sequencing of three genomic RNA segments of the Japanese and Malaysian Batai virus strains revealed that the M RNA segment of Batai virus had high sequence identity with that of Ngari virus. Our results indicate that Ngari virus is a genetic reassortant with S and L RNA segments from Bunyamwera virus and an M RNA segment from Batai virus.

The resurgence of Shamonda virus, an African Simbu group virus of the genus Orthobunyavirus , in Japan

Summary.Six virus isolations were made from Culicoides biting midges and blood samples of sentinel cattle in Kagoshima and Miyazaki Prefectures, the southern part of Japan, in 2002. Serological and genetical tests identified these viruses as isolates of Shamonda virus (SHAV), which belongs to the Simbu group of the genus Orthobunyavirus of the family Bunyaviridae. Initially, SHAV was isolated from cattle and Culicoides biting midges in Nigeria in the 1960s, and its presence has not been reported until this study. The present results indicate a wider distribution of SHAV than previously assumed.

Serological and genetic characterization of newly isolated Peaton virus in Japan

Summary. The viruses were isolated from the blood of sentinel cattle and Culicoides biting midges in the Kyushu district, southwestern Japan, in 1999 and identified by neutralization tests as Peaton (PEA) viruses. Before this study, PEA virus had been isolated in Australia only. The nucleotide identity of the nucleocapsid (N) protein encoded by the S segment ranged from 91.1 to 91.6% between the Australian and Japanese strains. A phylogenetic analysis of the N protein sequence revealed that the PEA virus strains are closely related to Aino (AIN) virus and suggested reassortment events for PEA and AIN viruses.

Genetic characterization of Aino and Peaton virus field isolates reveals a genetic reassortment between these viruses in nature

Sequence determination and phylogenetic analysis were conducted using the S, M and L RNA segments of the 10 Aino, 6 Peaton and 1 Sango virus (AINOV, PEAV and SANV) field isolates of the genus Orthobunyavirus in the family Bunyaviridae, respectively. The Japanese AINOV strains were genetically stable, but the sequence differences between the Japanese and Australian AINOV strains were considerably larger than those among the Japanese AINOV strains. A similar result was found in the genetic relationship among Japanese and Australian PEAVs, and SANV which was isolated in Nigeria and was thought as a synonym of PEAV, suggesting that geographic separation contributed significantly to the evolution of those viruses. The Australian AINOV strain B7974 is more closely related to the Australian PEAV strain CSIRO110 than to the Japanese AINOV strains in the S and L RNA segments, while the phylogenetic position of the M RNA segment of the B7974 strain was clustered with those of the Japanese AINOV strains. Our findings indicate that the B7974 strain is a reassortment with the M RNA segment derived from AINOV and the S and L RNA segments derived from an Australian PEAV.

Phylogenetic relationships of the G gene sequence of bovine ephemeral fever virus isolated in Japan, Taiwan and Australia.

The G gene encoding the neutralization antigen of bovine ephemeral fever virus (BEFV) was characterized in order to define the viruss molecular epidemiology in Japan and the genetic relationships among the Japanese, Taiwanese and Australian isolates. The nucleotide and amino acid sequences of the gene were highly conserved among the Japanese strains, regardless of the year of isolation, and were closely related to the Taiwanese strains. By phylogenetic analysis, the Japanese and Taiwanese strains were classified clearly into three chronological clusters: 1966, 1984-1989 and 1996-2004, indicating that the epidemics of bovine ephemeral fever may occur almost simultaneously in both countries by the same genotype. On the other hand, the Australian strains were distantly related to these East Asian strains and placed in the independent fourth cluster of the phylogenetic tree. It is suggested that three amino acid substitutions at residues 224, 271 and 499 in the neutralizing epitopes, of which two generate new glycosylation sequences, are responsible for antigenic variations of bovine ephemeral fever virus. The cross-neutralization test using the bovine ephemeral fever virus isolated in Japan demonstrated that the vaccine developed based on the oldest Japanese strain, YHL, appears to still be effective for controlling bovine ephemeral fever in Japan.

Sequence analysis of the medium RNA segment of three Simbu serogroup viruses, Akabane, Aino, and Peaton viruses ☆

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.

The emergence in Japan of Sathuperi virus, a tropical Simbu serogroup virus of the genus Orthobunyavirus

Summary.In 1999, two viruses were isolated from blood samples of sentinel cattle in the Western part of Japan. The physiochemical and morphological properties of these viruses indicated that they belonged to the family Bunyaviridae. Sequence analysis of the S segment indicates that the two viruses are closely related to Sathuperi virus (SATV). The N-terminal 168 amino acid of the G2 protein of the M segment was highly homologous with that of SATV (98.2%). Given these results, we conclude that the newly isolated viruses are closest to SATV, which was initially isolated in India and Nigeria over 30 years ago.