BACKGROUND:Nairobi sheep disease virus (NSDV), a highly pathogenic tick-borne orthonairovirus affecting ruminants, represents an emerging zoonotic threat. While endemic to East Africa, recent reports confirm its presence in Asia. HYPOTHESIS/OBJECTIVES:This study investigated NSDV circulation in the eastern Shandong Peninsula. ANIMALS:A total of 745 Haemaphysalis longicornis ticks (grouped into 139 pools) and 246 ruminant serum samples were collected during 2021 field surveillance in Weifang and Yantai regions. METHODS:Molecular detection employed quantitative reverse transcription PCR and nested PCR assays, with genomic characterization through complete and partial segment sequencing. Serological analysis used ELISA and immunofluorescence assays (IFAs). Phylogenetic reconstruction was performed using maximum-likelihood methods. RESULTS:Nairobi sheep disease virus RNA was detected in 9 tick pools (minimum infection rate: 1.2%). Genomic analysis revealed one complete genome (L: 11 999 bp; M: 5013 bp; and S: 1469 bp) and 4 partial S segment sequences (975 bp) showing > 98% nucleotide identity with Chinese reference strains and 100% identity among themselves. Phylogenetically, Shandong isolates clustered with other Chinese NSDV variants, indicating local evolutionary adaptation. Seroprevalence reached 5.7% (14/246) among ruminants, suggesting local virus exposure. CONCLUSIONS AND CLINICAL IMPORTANCE:This molecular and serological evidence of NSDV in eastern Shandong suggests the potential presence of the virus beyond its historical range. The close genetic relationship with other Chinese strains suggests regional spread rather than independent introductions. These findings underscore the need for enhanced surveillance in tick-ruminant systems and the development of control strategies to mitigate economic and zoonotic risks.

Arboviruses represent a persistent and escalating threat to global public health, with mosquitoes serving as the principal vectors in their natural transmission cycles and geographic dissemination. Yunnan Province, southwestern China, is a recognized hotspot for arboviral diversity, yet the full spectrum of mosquito-borne viruses circulating in this region remains incompletely characterized. A total of 3,300 female mosquitoes of four species across four genera were collected from rural areas of Honghe County, Yunnan Province in 2024, and subjected to viral metatranscriptomic sequencing. A previously undescribed bunya-like virus, designated Honghe Bunya-like virus, was identified in two locally dominant hematophagous mosquito species, with minimum infection rates of 0.2

La Crosse virus (LACV), a constituent of the California serogroup (CSG) within the genus Orthobunyavirus in the Peribunyaviridae family, is the causative agent of LACV encephalitis. This form of encephalitis stands as one of the most significant and burgeoning mosquito-borne diseases in the United States, ranking as the second most prevalent mosquito-borne illness following West Nile virus encephalitis. Predominantly identified in the Midwestern, Mid-Atlantic, and Southeastern regions of the United States, LACV primarily afflicts humans through the bites of Aedes triseriatus mosquitoes. Its genome, divided into three segments, encodes proteins that not only facilitate efficient replication within hosts but also hinder host immune responses. Infections by LACV can lead to a spectrum of neurological outcomes, ranging from mild aseptic meningitis to severe encephalitis with the potential for long-lasting neurological deficits. Despite the availability of diagnostic tools, several challenges persist. Currently, the management of LACV infection remains supportive, underscoring the importance of preventative measures in substantially mitigating the infection's incidence and severity. Moreover, global warming elevates the risk of LACV spreading to new territories. This review delves into recent advancements concerning the transmission and pathogenesis of LACV, drawing upon current knowledge regarding its genetic framework, transmission modes, geographical spread, phylogenetic relationships, clinical presentations and neuropathogenic effects, diagnostic approaches, treatment modalities, and prevention strategies.

Soft ticks, an important group of blood-sucking arthropods in nature, are widely distributed globally and can carry a wide range of pathogens, including Theileria ovis, Anaplasma ovis, Rickettsia spp. and African swine fever virus. In this study, we identified a novel single positive-stranded RNA virus, tentatively named Xiabuqu River virus (XRV), for the first time from soft ticks collected in Shigatse, Tibet. A total of 96 engorged soft ticks were collected from Tibetan sheep, with each tick assigned to a separate pool for analysis. The complete coding sequence of XRV was determined through next-generation sequencing, revealing a sequence length of 9277 nucleotides that includes a single open reading frame (ORF) encoding proteins such as the capsid protein, RNA helicase, and RNA-dependent RNA polymerase (RDRP). Quantitative RT-PCR and nested PCR were utilized to investigate the distribution of XRV within the tick sample. Pairwise distance analysis revealed that all obtained viral sequences shared a high nucleotide identity. Phylogenetic analysis demonstrated that XRV clustered with Lhasa Iflav tick virus 1, Fuyun tick virus 2, and Hubei tick virus 2. Further analyses indicated that XRV is a new member of the unclassified genus Iflavirus within the family Iflaviridae.

Arboviruses are a major public health concern. Wenzhou sobemo-like virus 4 (WZSLV4) is a recently identified single-stranded RNA mosquito-borne virus. In this study, 3608 mosquitoes were collected from Shandong Province, China, and divided into 58 pools according to species and sex. qRT-PCR and nested PCR were used to confirm the presence of WZSLV4. Cell culture was used to isolate the virus, and the growth characteristics of WZSLV4 were observed. WZSLV4 was detected in Aedes albopictus (12.07 %, 7/58) and Culex tritaeniorhynchus (18.97 %,11/58). WZSLV4 replicated effectively in C6/36 cells but grew slowly in Vero and BHK cells without causing cytopathic effect (CPE). Phylogenetic analysis demonstrated that WZSLV4 was closely related to the Guangzhou sobemo-like virus and Sichuan mosquito sobemo-like virus, which belongs to the family Solemoviridae. Pairwise distance analysis indicated that 11 partially amplified sequences shared a high nucleotide identity. The objective of this study was to isolate and characterize WZSLV4 from mosquito populations in Shandong Province, China, and to evaluate its growth characteristics in different cell lines to better understand its biological properties and potential pathogenicity. To our knowledge, this is the first report on the isolation of WZSLV4 from mosquitoes, along with a description of the biogenic characterization of WZSLV4 in different cell lines. Nevertheless, further studies are needed to assess the potential pathogenic risks in mammals and humans.

Arboviruses represent a growing threat to global public health, as climate change is exacerbating the spread of these vectors and thereby increasing the transmission potential of arboviruses. As primary vectors, mosquitoes harbor remarkably diverse virus populations, broadly classified into mosquito-borne viruses and mosquito-specific viruses. Although metagenomic sequencing has revolutionized viral discovery, geographical variations in mosquito viromes remain poorly characterized. In this study, we conducted comprehensive viral surveillance on 5,051 mosquitoes representing six medically important species (grouped into 14 taxonomic units) collected from Shandong, China, in 2021. Using next-generation sequencing, we generated 3.7 billion clean reads and identified 10 distinct virus species spanning eight virus families. Phylogenetic analyses revealed substantial genomic diversity and complex evolutionary relationships, including two completely novel virus species and several incompletely characterized known viruses. Notably, Culex quinquefasciatus mosquitoes exhibited significantly greater viral diversity than other species, although we observed considerable overlap in viral communities across mosquito taxa, suggesting non-host-specific viral maintenance. The presence of two novel viruses in field populations was confirmed by RT-qPCR screening, wherein minimum infection rates of 0.16% and 0.38% were established. Our findings substantially expand the known diversity of mosquito-associated viruses in East Asia and provide critical baseline data for arbovirus surveillance programs. This work highlights the value of systematic virome characterization for predicting emerging arboviral threats and understanding virus-mosquito ecology in changing environments. IMPORTANCE:Ten viruses, including two novel ones, were found in a study performed on mosquitoes in Shandong, China. It shows viral diversity and coexistence in different species, highlighting host impact on viral communities. The new viruses are prevalent locally, with infection rates of 0.38% and 0.16%. This work advances viral ecology understanding and has public health significance. This study sheds light on the circulation of the identified viruses in Shandong.

The Alongshan virus (ALSV), classified within the Flaviviridae family and belonging to the Jingmenvirus group, is a segmented RNA virus that was first identified in China in 2017. Since then, it has been reported in several Eurasian countries. Although no confirmed fatal cases have been documented, the potential public health risks associated with ALSV are significant and warrant serious attention. The emergence of ALSV has not only broadened the array of tick-borne diseases but has also enriched the research landscape surrounding segmented flaviviruses. Despite these advancements, our understanding of ALSV is still nascent, and its complex infection pathways remain largely unexplored. This review seeks to offer an in-depth examination of ALSV, addressing its biological properties, molecular features, epidemiological data, clinical presentations, and diagnostic methodologies. Our objective is to promote progress in the formulation of preventive, diagnostic, and therapeutic measures for this emerging segmented flavivirus.

Mosquitoes are important vectors of a wide range of viruses and serve as pivotal vectors with complex ecological functions that facilitate the transmission of these pathogens between hosts, such as humans, through biting behavior. In the mosquito surveillance conducted in Weifang and Yantai cities of Shandong Province, genetic material of a novel virus, temporarily named Fangshan bunya-like virus, was detected. From 3013 female mosquitoes, genetic material was detected in 9 pools (infection rate: 0.30%) using qRT-PCR and nested PCR. Analysis of the distances between pairs revealed a significant level of nucleotide similarity among these amplified sequences. Phylogenetic analysis placed Fangshan bunya-like virus within the Mobuvirus genus, Phenuiviridae family, clustering with Pyongtaek Culex Bunyavirus and indicating a common ancestor. The genomic characteristics and phylogenetic classification of Fangshan bunya-like virus offered valuable insights into the diversity and evolution of Phenuiviridae among mosquito populations, despite the challenge of not having isolated the live virus.

Mosquito-borne viruses represent a major global public health threat, with transmission dynamics governed by climatic, ecological, and anthropogenic factors. Yantai City, Shandong Province, situated in a warm-temperate monsoon climate zone, shares geographical and ecological characteristics with regions where mosquito-borne viruses are endemic, creating potential for virus introduction. We used metagenomics to systematically analyze viral communities in mosquitoes from the Yantai region. We collected 8,111 mosquitoes representing four genera and six species, with Culex being predominant (89.8%). High-throughput sequencing revealed 11 viral species spanning 9 families, including Peribunyaviridae and Picornaviridae. Notably, Serbia mononega-like virus 1 and Biggievirus Mos11 represent the first reports from China, with quantitative reverse transcription PCR revealing minimum infection rates of 0.34% and 0.68%, respectively. Phylogenetic analysis revealed close relationships to known viral strains, with several isolates potentially representing novel genera or species. Analysis revealed that Culex quinquefasciatus harbored the greatest viral diversity (five species), with significantly higher viral diversity in agricultural versus urban areas (P < 0.001). Several viruses demonstrated cross-species transmission potential, including Zhee mosquito virus, Zhejiang mosquito virus 3, and Culex tritaeniorhynchus rhabdovirus, all detected across multiple mosquito species. While most viruses appear mosquito-specific, several show close phylogenetic relationships to known pathogens, potentially posing public health risks warranting surveillance. This study addresses knowledge gaps regarding mosquito-borne viruses in the Bohai Rim region and provides a scientific foundation for regional viral surveillance and early warning systems.IMPORTANCEMosquito-borne viruses are a significant global health threat, with the potential to cause widespread disease outbreaks. This study investigated the viral diversity within mosquito populations in Yantai, China, and characterized the molecular features of two emerging RNA viruses. These findings highlight the remarkable viral diversity harbored by Culex mosquitoes and reveal higher viral diversity in agricultural areas compared to urban settings. Several identified viruses exhibit cross-species transmission potential and close phylogenetic relationships to known pathogens, suggesting that they may pose public health risks. Understanding these interactions is essential for predicting how environmental changes may affect virus transmission and the resilience of surveillance and control strategies.

Climate change is increasing the threat of arboviruses globally, amplifying their geographic distribution and virulence. This poses a grave risk to public health as mosquito-transmitted viral infections resurge and spread. Unbiased metagenomics plays a crucial role in identifying unknown pathogens and monitoring significant ones. Using this approach, a novel mosquito-associated virus, named Chijiahe virus, was discovered. We collected 2,910 mosquitoes and divided them into 65 pools based on species and collection site. To ascertain the existence of Chijiahe virus, we utilized nested PCR alongside real-time quantitative reverse transcription PCR (RT-qPCR). Further analysis of the virus's genomic characteristics and phylogenetic relationships revealed its detection in 10 out of 65 mosquito pools, yielding a minimum infection rate (MIR) of 0.34%. The virus genome consists of a 9,654-nucleotide single-stranded RNA, containing an open reading frame (ORF) that encodes essential proteins, including the capsid protein, RNA helicase, and RNA-dependent RNA polymerase (RdRp). Pairwise distance analysis revealed high nucleotide identity among all obtained sequences. Phylogenetic analysis demonstrated Chijiahe virus's close relationship to Arthropod iflavirus, a member of the family Iflaviridae. Additional analyses confirmed Chijiahe virus as a novel member of Iflaviridae.

Tahyna virus (TAHV), a member of the California serogroup of the genus Orthobunyavirus, is an arbovirus that is capable of causing a range of symptoms from mild febrile illnesses to severe neuroinvasive disease. It was first isolated in Europe and has a holarctic distribution, spanning Central Europe, Asia, and Africa. Central Europe has reported the highest reported human seropositivity rate, reaching 30

Arboviruses persist as a significant threat to global public and human health. The advent of macrogenomics technology has substantially expanded our understanding of mosquito-borne viruses by facilitating the direct identification and characterization of viruses from diverse environmental samples. In 2022, we conducted mosquito surveillance in the Shandong region of China and discovered a novel virus, tentatively named Gushan virus based on the site of discovery. In this study, a total of 3170 mosquitoes were collected and divided into 62 pools based on species and collection locations. Quantitative reverse transcription PCR (qRT-PCR) and nested PCR were employed to detect Gushan virus nucleic acids in all samples. The results revealed that 16 pools were positive, with a minimum infection rate (MIR) of 0.5% (16/3,170). Concurrently, we successfully obtained one complete genome sequence and six partial sequences. Nucleotide homology analysis demonstrated a high degree of sequence identity, indicating that Gushan virus may maintain a stable genetic profile in the region. Phylogenetic analyses revealed that Gushan virus is most closely related to Drosophila immigrans Nora virus of the family Noroviridae. Taxonomically, Gushan virus belongs to the order Picornavirales, family Noroviridae, and genus Orthonoravirus. This study contributes new insights into the diversity of mosquito-associated viruses.

Ticks are the second most important vectors of human diseases after mosquitoes. Hard ticks are more abundant and widespread than soft ticks, resulting in their greater involvement in diverse diseases. Consequently, most research on tick-borne pathogens has focused on hard ticks. In contrast, soft ticks, which comprise fewer species, have received less research attention. In this study, we identified a novel single-stranded RNA virus (tentatively named Gagosa Mountain virus) in Ornithodoros lahorensis ticks from the Shigatse region of Tibet. We collected 80 engorged soft ticks from Tibetan sheep, placing each in a separate tube for pathogen analysis. Quantitative real-time PCR (qRT-PCR) and nested PCR techniques were employed to confirm the presence of Gagosa Mountain virus, and subsequent analyses focused on elucidating its genomic features and phylogenetic relationships. Our results demonstrated that Gagosa Mountain virus was present in 15 out of 80 ticks, corresponding to a positivity rate of 19%. The 13,133-nucleotide single-stranded negative-sense RNA genome contained six open reading frames (ORFs) encoding the N protein, RdRp, and four hypothetical proteins. Pairwise distance analysis showed high nucleotide sequence identity among the viral sequences. Phylogenetic analysis indicated that Gagosa Mountain virus is most closely related to Lhasa Rhabd tick virus 1, which is an unclassified member of the family Rhabdoviridae. Further analyses demonstrated that Gagosa Mountain virus represents a novel member of the genus Betanemrhavirus within the family Rhabdoviridae.

Mosquito associated virus have always been a significant threat to global health. Metagenomics offers a straightforward and quantitative means to acquire the information of novel virus and has greatly enriched the content of mosquito associated virus databases. During an entomological surveillance for arthropod-borne viruses in China, we identified a previously unrecognized virus from mosquitoes, temporarily named Huajieling virus. In this study, a total of 3,960 mosquitoes were collected and then divided into 91 pools, according to location and species. QRT-PCR and nested PCR were performed to confirm the presence of Huajieling virus. Its genomic features and phylogenetic relationships were further analyzed. Our results showed that Huajieling virus was detected in 7 of the 91 mosquito pools and that the minimum infection rate (MIR) was 0.18% (7/3,960). One complete genome sequence and 2 viral partial sequences were obtained from the Huajieling virus-positive pools. Pairwise distances analysis indicated that these amplified sequences shared high nucleotide identity. Phylogenetic analysis demonstrated that Huajieling virus is most closely related to Wufeng shrew picorna-like virus 43, which belonging to Picornavirales. Further analyses indicated that Huajieling virus is a new member of unclassified Picornavirales, and is intermediate between the family Caliciviridae and Secoviridae in taxonomic status.

Jingmen tick virus (JMTV) is a novel segmented Flavivirus that was first identified from Rhipicephalus microplus in the Jingmen region of Hubei Province, China, in 2010. Subsequently, it was detected in a variety of countries and regions around the world. Meanwhile, JMTV has been proved to be pathogenic to humans and animals and could cause viremia in animals. However, the pathogenic mechanism of JMTV and what role animals play in the viral cycle have not yet been elucidated. In this study, 38 sheep sera were collected from Xinyang region of Henan Province, China and 204 ticks attached to the sheep were collected. The qRT-PCR and nested PCR were used to confirm the presence of JMTV in serum and tick samples. The results showed that the positive rate of JMTV in serum and ticks was 13.16% (5/38) and 7.84% (16/204), respectively. Phylogenetic analysis showed that JMTV sequences in sheep and ticks shared a high degree of identity with each other, and JMTV was relatively conserved in evolution. These results enriched the evidence for the prevalence of JMTV in animals and further deepened our understanding of the mechanisms and routes of JMTV transmission.

Mosquito associated virus have always been a significant threat to global health. Metagenomics offers a straightforward and quantitative means to acquire the information of novel virus and has greatly enriched the content of mosquito associated virus databases. During an entomological surveillance for arthropod-borne viruses in China, we identified a previously unrecognized virus from mosquitoes, temporarily named Huajieling virus. In this study, a total of 3,960 mosquitoes were collected and then divided into 91 pools, according to location and species. QRT-PCR and nested PCR were performed to confirm the presence of Huajieling virus. Its genomic features and phylogenetic relationships were further analyzed. Our results showed that Huajieling virus was detected in 7 of the 91 mosquito pools and that the minimum infection rate (MIR) was 0.18% (7/3,960). One complete genome sequence and 2 viral partial sequences were obtained from the Huajieling virus-positive pools. Pairwise distances analysis indicated that these amplified sequences shared high nucleotide identity. Phylogenetic analysis demonstrated that Huajieling virus is most closely related to Wufeng shrew picorna like virus 43, which belonging to Caliciviridae. Further analyses indicated that Huajieling virus is a new member of unclassified Caliciviridae.

Arboviruses have always been a significant public health concern. Metagenomic surveillance has expanded the number of novel, often unclassified arboviruses, especially mosquito-borne and mosquito-specific viruses. This report presents the first description of a novel single-stranded RNA virus, Wanghe virus, identified from mosquitoes that were collected in Shandong Province in 2022. In this study, a total of 4,795 mosquitoes were collected and then divided into 105 pools according to location and species. QRT-PCR and nested PCR were performed to confirm the presence of Wanghe virus, and its genomic features and phylogenetic relationships were further analyzed. Our results revealed that Wanghe virus was detected in 9 out of the 105 mosquito pools, resulting in a minimum infection rate (MIR) of 0.19 % (9/4,795). One complete genome sequence and three viral partial sequences were obtained from the Wanghe virus-positive pools. Pairwise distance analysis indicated that these amplified sequences shared high nucleotide identity. Phylogenetic analysis demonstrated that Wanghe virus is most closely related to Guiyang Solinvi-like virus 3, which belongs to Solinviviridae. Further analyses indicated that Wanghe virus is a new, unclassified member of Solinviviridae.

The Oropouche virus is an important arthropod-borne virus in the Peribunyaviridae family that can cause febrile illnesses, and it is widely distributed in tropical regions such as Central and South America. Since the virus was first identified, a large number of related cases are reported every year. No deaths have been reported to date, however, the virus can cause systemic infections, including the nervous and blood systems, leading to serious complications. The transmission of Oropouche virus occurs through both urban and sylvatic cycles, with the anthropophilic biting midge Culicoides paraensis serving as the primary vector in urban areas. Direct human-to-human transmission of Oropouche virus has not been observed. Oropouche virus consists of three segments, and the proteins encoded by the different segments enables the virus to replicate efficiently in the host and to resist the host's immune response. Phylogenetic analyses showed that Oropouche virus sequences are geographically distinct and have closer homologies with Iquitos virus and Perdoes virus, which belong to the family Peribunyaviridae. Despite the enormous threat it poses to public health, there are currently no licensed vaccines or specific antiviral treatments for the disease it causes. Recent studies have utilised imJatobal virusmunoinformatics approaches to develop epitope-based peptide vaccines, which have laid the groundwork for the clinical use of vaccines. The present review focuses on the structure, epidemiology, immunity and phylogeny of Oropouche virus, as well as the progress of vaccine development, thereby attracting wider attention and research, particularly with regard to potential vaccine programs.

Our study provided important epidemiological evidence for the epidemic of HMV2 in Shandong Province. Here, we report the first isolation and molecular characteristics of this virus and enrich our knowledge of the distribution of HMV2 in China.

目的 了解疫情风险对定向医学生履约意愿的影响,提出促进诚信履约的学校教育策略和措施.方法 通过问卷调查法搜集信息,采用回归分析和结构方程模型分析定向医学生履约意愿的影响因素及其作用路径.结果 描述性分析发现,女性履约意愿高于男性;预防医学专业定向医学生履约意愿相对较高,而精神医学专业定向医学生的履约意愿相对较低;随着年级的增加,定向医学生的履约意愿逐步降低.多重线性回归结果表明,在控制性别、年级、专业等因素的情况下,疫情风险感知(β=0.062,95%CI:0.007～0.117)和职业角色认同(β=0.283,95%CI:0.241～0.325)对履约意愿的影响均具有统计学意义.结构方程模型显示,疫情风险感知对履约意愿的影响不仅受到性别和年级的影响,而且职业角色认同是前者影响后者的重要中介.结论 加强定向医学生公共卫生知识与技能的培养,以增强其公共卫生处置能力;将职业生涯规划贯穿学校教育的全过程,增强职业认同意识,重点管控好高年级定向医学生的违约风险,提升其服务基层的履约信念.