Evolution and genetic characterization of Seoul virus in wild rats Rattus norvegicus from an urban park in Lyon, France 2020-2022.

BACKGROUND: Seoul virus (SEOV) is an orthohantavirus primarily carried by rats. In humans, it may cause hemorrhagic fever with renal syndrome (HFRS). Its incidence is likely underestimated and given the expansion of urban areas, a better knowledge of SEOV circulation in rat populations is called for. Beyond the need to improve human case detection, we need to deepen our comprehension of the ecological, epidemiological, and evolutionary processes involved in the transmission of SEOV. METHODOLOGY / PRINCIPAL FINDINGS: We performed a comprehensive serological and molecular characterization of SEOV in Rattus norvegicus in a popular urban park within a large city (Lyon, France) to provide essential information to design surveillance strategies regarding SEOV. We sampled rats within the urban park of 'La Tête d'Or' in Lyon city from 2020 to 2022. We combined rat population genetics, immunofluorescence assays, SEOV high-throughput sequencing (S, M, and L segments), and phylogenetic analyses. We found low structuring of wild rat populations within Lyon city. Only one sampling site within the park (building created in 2021) showed high genetic differentiation and deserves further attention. We confirmed the circulation of SEOV in rats from the park with high seroprevalence (17.2%) and high genetic similarity with the strain previously described in 2011 in Lyon city. CONCLUSION/SIGNIFICANCE: This study confirms the continuous circulation of SEOV in a popular urban park where the risk for SEOV transmission to humans is present. Implementing a surveillance of this virus could provide an efficient early warning system and help prepare risk-based interventions. As we reveal high gene flow between rat populations from the park and the rest of the city, we advocate for SEOV surveillance to be conducted at the scale of the entire city.

Seoul Virus Infection and Spread in United States Home-Based Ratteries: Rat and Human Testing Results From a Multistate Outbreak Investigation.

BACKGROUND: During 2017, a multistate outbreak investigation occurred after the confirmation of Seoul virus (SEOV) infections in people and pet rats. A total of 147 humans and 897 rats were tested. METHODS: In addition to immunoglobulin (Ig)G and IgM serology and traditional reverse-transcription polymerase chain reaction (RT-PCR), novel quantitative RT-PCR primers/probe were developed, and whole genome sequencing was performed. RESULTS: Seventeen people had SEOV IgM, indicating recent infection; 7 reported symptoms and 3 were hospitalized. All patients recovered. Thirty-one facilities in 11 US states had SEOV infection, and among those with ≥10 rats tested, rat IgG prevalence ranged 2%-70% and SEOV RT-PCR positivity ranged 0%-70%. Human laboratory-confirmed cases were significantly associated with rat IgG positivity and RT-PCR positivity (P = .03 and P = .006, respectively). Genomic sequencing identified >99.5% homology between SEOV sequences in this outbreak, and these were >99% identical to SEOV associated with previous pet rat infections in England, the Netherlands, and France. Frequent trade of rats between home-based ratteries contributed to transmission of SEOV between facilities. CONCLUSIONS: Pet rat owners, breeders, and the healthcare and public health community should be aware and take steps to prevent SEOV transmission in pet rats and to humans. Biosecurity measures and diagnostic testing can prevent further infections.

Genetic Characterization and Molecular Evolution of Urban Seoul Virus in Southern China.

Seoul virus (SEOV), which causes hemorrhagic fever with renal syndrome (HFRS) in humans, has spread all over the world, especially in mainland China. Understanding basic mechanisms of SEOV evolution is essential to better combat and prevent viral diseases. Here, we examined SEOV prevalence and evolution in the residential area of four districts in Guangzhou city, China. The carriage of SEOV was observed in 33.33% of the sampled rodents, with 35.96% of the sampled Rattus norvegicus and 13.33% of R. tanezumi. Based on the comprehensive analyses of large (L), medium (M), and small (S) segments, our study first demonstrated that the genetic characterization of urban SEOV was shaped by high nucleotide substitution rates, purifying selection, and recombination. Additionally, we detected mutational saturation in the S segment of SEOV, which may lead to the biases of genetic divergence and substitution rates in our study. Importantly, we have filled the gap of SEOV evolution in the urban area. The genetic variation of SEOV may highlight the risk of HFRS, which merits further investigation.

Genetic analyses of Seoul hantavirus genome recovered from rats (Rattus norvegicus) in the Netherlands unveils diverse routes of spread into Europe.

Seoul virus (SEOV) is the etiologic agent of hemorrhagic fever with renal syndrome. It is carried by brown rats (Rattus norvegicus), a commensal rodent that closely cohabitates with humans in urban environments. SEOV has a worldwide distribution, and in Europe, it has been found in rats in UK, France, Sweden, and Belgium, and human cases of SEOV infection have been reported in Germany, UK, France, and Belgium. In the search of hantaviruses in brown rats from the Netherlands, we found both serological and genetic evidence for the presence of SEOV in the local wild rat population. To further decipher the relationship with other SEOV variants globally, the complete genome of SEOV in the Netherlands was recovered. SEOV sequences obtained from three positive rats (captured at close trapping locations at the same time) were found highly similar. Phylogenetic analyses demonstrated that two lineages of SEOV circulate in Europe. Strains from the Netherlands and UK, together with the Baxter strain from US, constitute one of these two, while the second includes strains from Europe and Asia. Our results support a hypothesis of diverse routes of SEOV spread into Europe. These findings, combined with other indications on the expansion of the spatial European range of SEOV, suggest an increased risk of this virus for the public health, highlighting the need for increased surveillance.

Genetic characterization of a hantavirus isolated from Heilongjiang province, China.

Hantavirus is the causative agent of hemorrhagic fever with renal syndrome (HFRS). Heilongjiang Province is experiencing an epidemic of HFRS, the main causative agent is a variant of hantavirus called Seoul virus (SEOV). In this study, the entire genome of one SEOV, the DN2 strain, was sequenced and analyzed. The alignment analysis of the sequences indicated that the DN2 strain shares the highest homology with the SEOV-LYO852 strain. The nucleotide identity is 97.6% for the S segment, 97.7% for the M segment, and 98.0% for the L segment. The corresponding amino acid sequence homologies are 99.1%, 98.9% and 99.8%. The phylogenetic analysis of the segments suggests that the DN2 strain has a high genetic relationship with SEOV strains and no genetic recombination occurs.

L'Hantavirus est l'agent causal de la fièvre hémorragique avec syndrome rénal (FHSR). La province d'Heilongjiang est au prise avec une épidémie de FHSR, l'agent causal principal est un variant de l'Hantavirus dénommé virus Séoul (SEOV). Dans la présente étude, les séquences complètes d'un SEOV, la souche DN2, ont été séquencées et analysées. L'analyse d'appariement des séquences a démontré que la souche DN2 partage la plus forte homologie avec la souche SEOV-LYO852. L'identité de nucléotides est de 97,6 % pour le segment S, 97,7 % pour le segment M, et de 98,0 % pour le segment L. L'homologie des séquences d'acides aminés correspondants est de 99,1 %, 98,9 %, et 99,8 %. L'analyse phylogénétique des segments suggéraient que la souche DN2 avait une parenté génétique la plus élevée avec les souches de SEOV et qu'aucune recombinaison génétique ne s'est produite.(Traduit par Docteur Serge Messier).

Autochthonous Human Case of Seoul Virus Infection, the Netherlands.

Orthohantaviruses are a group of rodentborne viruses with a worldwide distribution. The orthohantavirus Seoul virus (SEOV) can cause hemorrhagic fever with renal syndrome in humans and is distributed worldwide, like its reservoir host, the rat. Cases of SEOV in wild and pet rats have been described in several countries, and human cases have been reported in the United Kingdom, France, Canada, and the United States. In the Netherlands, SEOV has previously been found in wild brown rats. We describe an autochthonous human case of SEOV infection in the Netherlands. This patient had nonspecific clinical symptoms of an orthohantavirus infection (gastrointestinal symptoms and distinct elevation of liver enzymes). Subsequent source investigation revealed 2 potential sources, the patient's feeder rats and a feeder rat farm. At both sources, a high prevalence of SEOV was found in the rats. The virus closely resembled the Cherwell and Turckheim SEOV strains that were previously found in Europe.

Importation of Human Seoul Virus Infection to Germany from Indonesia.

Seoul hantavirus-associated hemorrhagic fever with renal syndrome cases are rare outside Asia and have not yet been found in Germany. We report clinical and molecular evidence for a Seoul virus infection in a patient in Germany. The infection was most likely acquired during a stay in Sulawesi, Indonesia.

Genetic characterization of hantaviruses isolated from rodents in the port cities of Heilongjiang, China, in 2014.

BACKGROUND: Hantavirus is a tripartite negative-sense RNA virus. It can infect humans through contaminated rodent excreta and causes two types of fatal human diseases: hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS). China exhibits the highest HFRS occurrence rate in the world, and the Heilongjiang area is one of the most severely infected regions. RESULTS: To obtain additional insights into the genetic characteristics of hantaviruses in the port cities of the Heilongjiang area in China, a molecular epidemiological investigation of hantaviruses isolated from rodents was performed in 2014. A total of 649 rodents (11 murine species and 1 shrew species) were caught in 12 port cities in Heilongjiang. Among these rodents, the most common species was A. agrarius, and the second-most common was R. norvegicus. A viral gene PCR assay revealed the presence of two specific genotypes of hantavirus, referred to as Hantaan virus (HTNV) and Seoul virus (SEOV), and the positive SEOV infection rate was higher than that for HTNV. A genetic analysis based on partial M segment sequences indicated that all of the isolates belonging to SEOV could be assigned to two genetic lineages, whereas the isolate belonging to HTNV could be assigned to only one genetic lineage. CONCLUSIONS: These results suggested that HTNV and SEOV are circulating in A. agrarius and R. norvegicus in the port cities in the area of Heilongjiang, but SEOV may be the dominant common hantavirus.

Old World hantaviruses in rodents in New Orleans, Louisiana.

Seoul virus, an Old World hantavirus, is maintained in brown rats and causes a mild form of hemorrhagic fever with renal syndrome (HFRS) in humans. We captured rodents in New Orleans, Louisiana and tested them for the presence of Old World hantaviruses by reverse transcription polymerase chain reaction (RT-PCR) with sequencing, cell culture, and electron microscopy; 6 (3.4%) of 178 rodents captured--all brown rats--were positive for a Seoul virus variant previously coined Tchoupitoulas virus, which was noted in rodents in New Orleans in the 1980s. The finding of Tchoupitoulas virus in New Orleans over 25 years since its first discovery suggests stable endemicity in the city. Although the degree to which this virus causes human infection and disease remains unknown, repeated demonstration of Seoul virus in rodent populations, recent cases of laboratory-confirmed HFRS in some US cities, and a possible link with hypertensive renal disease warrant additional investigation in both rodents and humans.

A survey of rodent-borne pathogens carried by wild Rattus spp. in Northern Vietnam.

To examine the prevalence of human pathogens carried by rats in urban areas in Hanoi and Hai Phong, Vietnam, we live-trapped 100 rats in January 2011 and screened them for a panel of bacteria and viruses. Antibodies against Leptospira interrogans (22·0%), Seoul virus (14·0%) and rat hepatitis E virus (23·0%) were detected in rats, but antibodies against Yersinia pestis were not detected. Antibodies against L. interrogans and Seoul virus were found only in adult rats. In contrast, antibodies to rat hepatitis E virus were also found in juvenile and sub-adult rats, indicating that the transmission mode of rat hepatitis E virus is different from that of L. interrogans and Seoul virus. Moreover, phylogenetic analyses of the S and M segments of Seoul viruses found in Rattus norvegicus showed that Seoul viruses from Hai Phong and Hanoi formed different clades. Human exposure to these pathogens has become a significant public health concern.

Dynamics of hantavirus infections in humans and animals in Wuhan city, Hubei, China.

Hemorrhagic fever with renal syndrome (HFRS) has been a significant public problem since the first cases were reported in 1961 in Wuhan city (capital of Hubei province of China). Epidemiological surveys were carried out to better understand the dynamics of hantavirus infection in humans and animals in Wuhan. During 1961-2011, a total of 21,820 HFRS cases were registered in Wuhan. The two large epidemics had occurred during 1970-1991. They reached peaks in 1973 and 1983, respectively. There have been <10 cases since 2005. The disease occurred in the whole region including the downtown areas, but mainly in two districts. Although in 1980s and 1990s HFRS cases mainly recorded in August and winter, since 2000 the disease has mainly occurred in spring and summer. In this study, hantaviruses were identified in Apodemus mice, Rattus rats, and Mus mice by indirect immunofluorescent-assay and RT-PCR. Serological and genetic analyses showed that Hantaan virus (HTNV) and Seoul virus (SEOV) co-circulated in rodents. Phylogenetic analysis of hantaviral genome sequences revealed a novel genetic lineage of HTNV circulating in rodents in Wuhan. Another lineage of HTNV was closely related to the lineages from the provinces located in the origin and delta of Yangtze River. Remarkably, SEOV variants identified in Wuhan were more closely related to the variants found outside China. Results of the present study showed that HFRS cases in Wuhan are caused by HTNV and SEOV. Phylogenetic analysis of the hantavirus sequences revealed that a novel genetic lineage of HTNV is present in rodents in Wuhan.

Genetic characterization of seoul hantavirus originated from norway rats (Rattus norvegicus) captured in Belgium.

Hantaviruses (genus Hantavirus, family Bunyaviridae) cause hemorrhagic fever with renal syndrome (HFRS) in Eurasia and hantavirus (cardio)pulmonary syndrome (HCPS) in the Americas. So far, in Europe, four pathogenic hantaviruses have been found, often in co-circulation: Puumala virus (PUUV), Dobrava virus (DOBV), Saaremaa virus (SAAV), and Seoul virus (SEOV). Of those, only PUUV was found in Belgium. Recently, in our search for hantaviruses in the Flanders region of Belgium we collected genetic and serological evidence for the presence of SEOV virus in local brown rats. In this article, the results of (phylo)genetic analysis of wild-type SEOV strain from the Flanders are presented. The analysis based on the complete S segment sequence and also partial M- and L-segment sequences revealed that the Belgian SEOV strain was related most closely to strains from France, Indonesia, Japan, Singapore, Cambodia (those associated with the species Rattus norvegicus) and Vietnam. Such a clustering was in perfect agreement with the results of direct sequence comparison and suggested the same evolutionary history for all three genome segments of the Belgian SEOV strain (i.e., no reassortment of genome segments). So far, SEOV has been found in two European countries, France and Belgium, and there is every reason to believe that the area of the virus distribution in Europe is not restricted to those countries.

Migration of Norway rats resulted in the worldwide distribution of Seoul hantavirus today.

Despite the worldwide distribution, most of the known Seoul viruses (SEOV) are closely related to each other. In this study, the M and the S segment sequences of SEOV were recovered from 130 lung tissue samples (mostly of Norway rats) and from six patient serum samples by reverse transcription-PCR. Genetic analysis revealed that all sequences belong to SEOV and represent 136 novel strains. Phylogenetic analysis of all available M and S segment sequences of SEOV, including 136 novel Chinese strains, revealed four distinct groups. All non-Chinese SEOV strains and most of the Chinese variants fell into the phylogroup A, while the Chinese strains originating from mountainous areas clustered into three other distinct groups (B, C, and D). We estimated that phylogroup A viruses may have arisen only within the last several centuries. All non-Chinese variants appeared to be directly originated from China. Thus, phylogroup A viruses distributed worldwide may share a recent ancestor, whereas SEOV seems to be as diversified genetically as other hantaviruses. In addition, all available mitochondrial DNA (mtDNA) sequences of Norway rats, including our 44 newly recovered mtDNA sequences, were divided into two phylogenetic groups. The first group, which is associated with the group A SEOV variants, included most of rats from China and also all non-Chinese rats, while the second group consisted of a few rats originating only from mountain areas in China. We hypothesize that an ancestor of phylogroup A SEOV variants was first exported from China to Europe and then spread through the New World following the migration of Norway rats.

Genetic analysis of hantaviruses and their rodent hosts in central-south China.

Hantaan virus (HTNV) and Seoul virus (SEOV) are two major zoonotic pathogens of hemorrhagic fever with renal syndrome (HFRS) in Asia. Hubei province, which is located in the central-south China, had been one of the most severe epidemic areas of HFRS. To investigate phylogenetic relationships, genetic diversity and geographic distribution of HTNV and SEOV in their reservoir hosts, a total of 687 rodents were trapped in this area between 2000 and 2009. Sequences of partial S- and M-segments of hantaviruses and mitochondrial D-loop gene from 30 positive samples were determined. Our data indicated that SEOV and HTNV were co-circulating in Hubei. Phylogenetic analysis based on partial S- and M-segment sequences revealed two and three previously undefined lineages of SEOV, and a novel genetic lineage of HTNV, respectively. Four inter-lineage reassortment SEOVs carried by Rattus norvegicus and Apodemus agrarius were observed. It suggests that SEOV may cause spillover infections to A. agrarius naturally. The abundance of the phylogenetic lineages of SEOV suggested that central-south China was a radiation center for SEOVs.

Geo-spatial hotspots of hemorrhagic fever with renal syndrome and genetic characterization of Seoul variants in Beijing, China.

BACKGROUND: Hemorrhagic fever with renal syndrome (HFRS) is highly endemic in mainland China, and has extended from rural areas to cities recently. Beijing metropolis is a novel affected region, where the HFRS incidence seems to be diverse from place to place. METHODOLOGY/PRINCIPAL FINDINGS: The spatial scan analysis based on geographical information system (GIS) identified three geo-spatial "hotspots" of HFRS in Beijing when the passive surveillance data from 2004 to 2006 were used. The Relative Risk (RR) of the three "hotspots" was 5.45, 3.57 and 3.30, respectively. The Phylogenetic analysis based on entire coding region sequence of S segment and partial L segment sequence of Seoul virus (SEOV) revealed that the SEOV strains circulating in Beijing could be classified into at least three lineages regardless of their host origins. Two potential recombination events that happened in lineage #1 were detected and supported by comparative phylogenetic analysis. The SEOV strains in different lineages and strains with distinct special amino acid substitutions for N protein were partially associated with different spatial clustered areas of HFRS. CONCLUSION/SIGNIFICANCE: Hotspots of HFRS were found in Beijing, a novel endemic region, where intervention should be enhanced. Our data suggested that the genetic variation and recombination of SEOV strains was related to the high risk areas of HFRS, which merited further investigation.

[Genetic analysis of hantaviruses carried by Rattus norvegicus collected from Henan and Neimenggu provinces, China].

OBJECTIVE: Genetic analysis was performed to infer the relationship between hantaviruses carried by Rattus norvegicus from Henan and Neimenggu provinces and the other known hantavirus and the vaccine strain. METHODS: Total RNA was extracted from lung tissues with Trizol reagent. The complete M and S segment sequences of strains NM133 and Q12 were amplified by RT-PCR. The purified DNA fragments were directly subjected to sequencing, and then to sequence analysis and phylogenetic analysis. RESULTS: The complete S segment sequences of strains NM133 and Q12 were found to be 1770 nt and 1772 nt in length respectively, with one open reading frame encoding 429 amino acids. The complete M segment sequences of both two strains are 3654 nucleotide in length encoding a protein of 1133 amino acids. The two strains shared a high degree of homology with most of known Seoul virus (SEOV) but quite different from Hantaan virus and other hantaviruses. Furthermore, the nucleoprotein and glycoprotein of the two strains had the congruent structure with the vaccine strain Z37. On the S- and M-phylogenetic trees, both strains (NM133 and Q12) were grouped into the first cluster of SEOV, and were more closely related to the strains, such as: Hb8610, R22, HB55, L99, and K24-e7. CONCLUSION: Both strains (NM133 and Q12) belonged to SEOV, and sharing a high degree of homology and similar secondary structure with strains including the vaccine strains Z37, our data suggested that the present vaccine used in China could effectively prevent HFRS caused by SEOV.

[Survey on host animal and molecular epidemiology of hantavirus in Chuxiong prefecture, Yunnan province].

OBJECTIVE: To determine the hosts of hantavirus (HV) and its molecular epidemiological characteristics, to provide evidence for prevention and control on hemorrhagic fever with renal syndrome (HFRS). METHODS: Rodents were captured by a special trap within the residential area. The antigens of HV in lung tissues were detected by direct immuno-fluorescence assay (DFA). Nucleotide sequences of HV were amplified by RT-PCR with HV genotype-specific primer. The amplified genes were then sequenced. Phylogenetic tree were built on nucleotide sequence with ClustalX 1.83 software. RESULTS: 1421 rodents were captured and classified into 8 species of 4 Genera in the epidemic area within 10 counties of Chuxiong prefecture, Yunnan province, between 2005 and 2006. Out of the 1421 rodents, 1056 (74.31% ) of them were Rattus norvegicus and 280 (19.70%) belonged to Rattus flavipectus. The antigens of HV were detected by DFA in lung tissues and the total positive rate of HV was 5.15% (53/ 1029). After applying the sequencing nucleotide method to the 53 positive specimens, data showed that 21 specimens were positive and all of them belonged to Seoul type (15 samples were from Rattus norvegicus, 4 samples Rattus flavipectus, 2 samples Rattus nitidus). The partial S segments from 12 specimens were sequenced which appeared homologic with R22, L99 and HLD65 from GenBank in relatively high level (87.1% -99.7%). When compared to 76-118 strain of Hantaan type, their homologic degree was only 64.4%-69.1%. Results from Phylogenetic analysis showed that 12 specimens belonged to Seoul type. As for their homology, they were significantly similar to Seoul type and could be tentatively divided into two subtypes S1 and S3. CONCLUSION: It was confirmed that the Seoul type virus, as HFRS's pathogenetic agent mainly carried by rats, prevailed widely in Chuxiong prefecture. Owing to the local ecological environment, we also noticed the characteristics of different HV subtypes among Seoul type.

Hantaviruses in rodents and humans, Inner Mongolia Autonomous Region, China.

Surveys were carried out in 2003-2006 to better understand the epidemiology of hantaviruses in the Inner Mongolia Autonomous Region of China (Inner Mongolia). Hemorrhagic fever with renal syndrome (HFRS) was first reported in this region in 1955 and has been an important public health problem here since then. During 1955-2006, 8,309 persons with HFRS were reported in Inner Mongolia (average incidence rate 0.89/100,000), and 261 (3.14%) died. Before the 1990s, all HFRS cases occurred in northeastern Inner Mongolia. Subsequently, HFRS cases were registered in central (1995) and western (1999) Inner Mongolia. In this study, hantaviral antigens were identified in striped field mice (Apodemus agrarius) from northeastern Inner Mongolia and in Norway rats (Rattus norvegicus) from middle and western Inner Mongolia. Phylogenetic analysis of hantaviral genome sequences suggests that HFRS has been caused mainly by Hantaan virus in northeastern Inner Mongolia and by Seoul virus in central and western Inner Mongolia.

Genotyping of hantaviruses occurring in Linyi, China, by nested RT-PCR combined with single-strand conformation polymorphism analysis.

The hemorrhagic fever with renal syndrome (HFRS) incidence rate still holds in high level in some areas of the world, but a reliable and simple typing method that can be used in clinical diagnosis and epidemiologic surveys is not available. In this study, 48 serum samples were collected from patients with HFRS in Linyi area, China, that is seriously affected by this disease. The collected samples were analyzed by nested RT-PCR combined with single-strand conformation polymorphism (SSCP) for genotyping of hantaviruses (HV). Out of 48 serum samples, 41 samples were positive by the nested RT-PCR. According to the SSCP patterns, Seoul virus (SEOV) was found in 33 samples and Hantaan virus (HTNV) in 8 samples. The comparison of sequence identities of nested RT-PCR products of tested samples with reference isolates SEOV and HTNV supported the typing results. Thus, genotyping of HV by nested RT-PCR/SSCP is suitable in early diagnosis of HV infection and in epidemiologic surveys.