Seroepidemiological investigation of Getah virus in the China-Myanmar border area from 2022-2023.

Getah Virus (GETV) is an RNA virus that is transmitted by mosquitoes and can cause disease or death in a variety of vertebrates. Its prevalence is increasingly severe in Asia. This study conducted a GETV epidemiological investigation on 1,300 bovine sera collected in the Honghe Prefecture of Yunnan Province on the China-Myanmar border from 2022 to 2023. The positive rate of GETV antibodies in bovine serum in Honghe Prefecture was determined to be 20.25% through indirect Enzyme-linked immunosorbent test (ELISA) methods. Using Real-time PCR methods to detect GETV RNA in bovine serum, the positive rate was 0.23% (3/1300), and viral nucleic acids were only detected in three bovine sera in Jianshui area in 2022. The YN2305 strain was successfully isolated from mouse neuroblastoma (N2a) cells and the complete gene sequence was obtained. All the above results indicate the existence of GETV infection in cattle in Honghe Prefecture, Yunnan Province. Homology and genetic evolution analysis found that the isolated strain has a high homology with the JL1808 strain isolated from cattle in 2018, with a nucleotide identity of 100%, and a nucleotide identity of 99.8% with the SD17-09 strain isolated from foxes in 2017. Compared with the nucleotides of 44 virus strains published in Genbank, YN2305 has multiple nucleotide site mutations in the structural gene E2 and non-structural gene NS. The nucleotide and amino acid identity of the E2 gene are 94.2-100% and 96.4-100%, respectively. Genetic evolution analysis found that this virus strain is most closely related to the bovine origin JL1808, and it is in gene group III with HuN1, Kochi-01, SD17-09, MI-110-C1, and MI-110-C2 strains that causes significant clinical symptoms in Chinese pig, fox and horse populations, belonging to the same evolutionary branch. This study determined the infection rate, genotype, and main prevalence areas of GETV in bovine sera in the China-Myanmar border area. Therefore, the epidemiological investigation of GETV infection in multiple animal hosts should be further expanded, and research on its pathogenicity and vectors should be carried out.

Molecular survey of duck circovirus infection in poultry in southern and southwestern China during 2018 and 2019.

BACKGROUND: Duck circovirus (DuCV) is a potential immunosuppressive virus that causes feather disorders in young ducks. In this study, DuCV obtained from various species of ducks was investigated by polymerase chain reaction (PCR) in southern and southwestern China (Guangdong, Guangxi and Yunnan provinces) from 2018 to 2019. RESULTS: A total of 848 bursa samples were collected from dead Mulard, Cherry Valley Pekin, Muscovy and Mallard ducks from duck farms. The positivity rate of DuCV in the total sample was approximately 36.91%. We found that the prevalence of DuCV in Yunnan (43.09%) was higher than those in Guangxi (34.38%) and Guangdong (34.4%). However, the positivity rates of DuCV in the four duck species were not significantly different (P > 0.05). Nineteen randomly selected complete viral genomes were sequenced. The complete genomes of the DuCV were 1987 to 1995 nt in length, and were 81.7-99.3% homologous to the other 57 sequences in GenBank. Phylogenetic analyses based on the complete genomes of 76 DuCVs showed that the 19 novel DuCV sequences from Guangdong and Guangxi provinces mainly belonged to the DuCV-1 and DuCV-2 genetic groups, respectively. However, the two genotype groups coexisted in Yunnan Province. In addition, recombination analysis showed putative recombination sites in 3 strains in Yunnan that originated from strains Guangdong and Guangxi. Interestingly, the epidemiological investigation showed that Mulard ducks, Cherry Valley Pekin ducks and Muscovy ducks more than 4 weeks old were more susceptible to infection with the novel DuCV than ducks less than 4 weeks old. CONCLUSIONS: These data provide insight into the molecular epidemiology and genetic diversity of DuCVs circulating in southern and southwestern China for the first time.

First isolation and characterization of Getah virus from cattle in northeastern China.

BACKGROUND: Getah virus (GETV) is a neglected mosquito-borne Alphavirus that causes pyrexia, body rash, and leg oedema in horses and foetal death and reproductive disorders in pigs. Infected animals may play a critical role in the amplification and circulation of the virus. The present study aimed to investigate GETV infection in clinically infected cattle and vector mosquito species in northeastern China. RESULTS: Serum samples were collected from beef cattle that presented sudden onset of fever in forest grazing areas, and metagenomic sequencing was conducted, revealing 29 contigs from ten serum samples matching the GETV genome. Quantitative RT-PCR (RT-qPCR) was performed with GETV RNA from 48 beef cattle serum samples, showing that the overall prevalence of GETV in the beef cattle samples was 6.25% (3/48). Serological investigation indicated that GETV neutralizing antibodies were detected in 83.3% (40/48, 95% CI 67-100) of samples from the study region. The GETV JL1808 strain was isolated from clinically infected cattle showing fever. Sequence comparisons showed high identity with the HuN1 strain, a highly pathogenic swine epidemic isolate obtained in Hunan province in 2017, at the nucleotide level (99.5%) and at the deduced amino acid level (99.7-99.9%). The phylogenetic analysis of JL1808 clustered in Group III, and also revealed a close genetic relationship with the HuN1 strain. Additionally, about 12,000 mosquitoes were trapped in this region. The presence of GETV infection was detected in mosquitoes, suggesting that the minimum infection rate (MIR) was 1.50‰, with MIRs of 1.67‰ in Culex pseudovishnui, 1.60‰ in Culex tritaeniorhynchus, and 1.21‰ in Anopheles sinensis. CONCLUSIONS: To the best of our knowledge, this is the first report of GETV infection in cattle. These results demonstrated that a highly pathogenic, mosquito-borne swine GETV can infect and circulate in cattle, implying that it is necessary to conduct surveillance of GETV infection in animals in northeastern China.

Rapid Visual Detection of Getah Virus Using a Loop-Mediated Isothermal Amplification Method.

Getah virus (GETV) is a mosquito-borne alphavirus that is considered to be an emerging pathogen. To date, reverse transcription loop-mediated isothermal amplification (RT-LAMP) has not been used to detect GETV. Therefore, we describe a novel, fast, and sensitive LAMP method to detect GETV. Amplification of GETV RNA can be obtained within 50 min at 65°C. This RT-LAMP method was verified to be highly specific for GETV, with no cross detection of other viruses. The assay was 103 and 101 times more sensitive than RT-PCR and RT-qPCR, respectively, for the detection of GETV RNA. This novel RT-LAMP method provides a practical and economical alternative for detecting GETV in mosquitoes and serum samples that can be used even in the field.

Comparison of transcriptome under red and blue light culture of Saccharina japonica (Phaeophyceae).

Saccharina japonica is one of the most important economic seaweeds. Several aspects such as photosynthesis in Saccharina lives are affected by blue light, the predominant light spectrum in the habitat. In this study, transcriptome profiling of S. japonica by next generation sequencing technology generated 55,102 qualified transcripts and 40.5 % transcripts were assigned to functional annotation. Expression of a large proportion of genes has been previously reported to be regulated by blue light, taking dark as control. However, by comparison among white, blue and red light, the significantly differentially expressed gene tags (DEGs) accounted for only 6.75 % of the identified sequences. It indicated that light-regulated gene expression in kelps is not a specific blue-light response. Unexpectedly, red light had more extensive effects on the transcriptomic activity than blue light did, since the most (68.4 %) DEGs were red light-regulated and only 17.5 % were specifically regulated by blue light. Some of the DEGs with the highest mRNA levels under blue light are not blue light-upregulated but red light-downregulated. The extensive regulation on gene expression under red light together with the abundant presence of phytochrome-like protein gene tags in S. japonica indicated their significant roles in the lives of brown algae. By highlighting the photosynthetic metabolism, blue light is more efficient than red light in triggering the pigment biosynthesis, light reaction and carbon fixation, revealing a molecular basis for rapid growth of kelps, since most of the time blue light is predominant in their habitat.

Genetic characterization of Toxoplasma gondii isolates from pigs in southwestern China.

The genetic diversity of Toxoplasma gondii varies in different geographical regions. Isolates of T. gondii in South America, for example, are genetically and biologically divergent from those in North America and Europe, where the population structure is highly clonal and composed mainly of 3 distinct lineages, i.e., Types I, II, and III. However, little is known of the T. gondii genotypes in the People's Republic of China. Toxoplasma gondii infection in pigs causes significant economic loss and presents a risk for human infection. We conducted a survey to determine the genetic diversity of this parasite in slaughtered pigs from Yunnan Province, southwestern China. In total, 412 DNA samples were extracted from hilar lymph nodes and livers of pigs from slaughterhouses in Yunnan Province in southwest China, 56 of which were found to be positive for the T. gondii SAG3 gene. These positive DNA samples were typed at 10 genetic markers, including 9 nuclear loci, i.e., SAG1, SAG2, SAG3, BTUB, GRA6, L358, PK1, c22-8, c29-2, and an apicoplast locus Apico. Of these, 5 isolates were genotyped with complete data for all loci. Only 1 genotype (ToxoDB 9) was identified, previously reported as a widespread lineage from pigs, cats, and human patients in China. The results indicate that this genotype may be the major T. gondii lineage in China and possibly all of eastern Asia. This is the first report of genetic typing of T. gondii isolates from pigs in China's southwestern Yunnan Province, the results of which have implications for the prevention and control of T. gondii infections in humans and other animals.

Seroprevalence of Toxoplasma gondii in pigs in southwestern China.

The seroprevalence of toxoplasmosis in pigs was investigated in Yunnan province, Southwestern China between March 2008 and January 2009. A total of 831 serum samples were collected from 9 counties and assayed for Toxoplasma gondii antibodies by indirect haemagglutination (IHA) test. Antibodies to T. gondii were found in 16.97% (141/831) with slaughter pigs having the highest rate (22.28%), followed by breeding sows (16.59%). The results of the present survey indicated that infection with T. gondii in pigs is widely spread in China, including the Yunnan province, and is of public health concern.