Molecular analysis of Gyrovirus galga1 variants identified from the sera of dogs and cats in China.

Gyrovirus galga1 (GyVg1), a member of the Anelloviridae family and Gyrovirus genus, has been detected in chicken and human tissue samples. In this study, the DNA of GyVg1-related gyroviruses in the sera of six dogs and three cats from Central and Eastern China was identified using PCR. Alignment analysis between the nine obtained and reference GyVg1 strains revealed that the genome identity ranged from 99.20% (DOG03 and DOG04 strains) to 96.17% (DOG01 and DOG06 strains). Six recombination events were predicted in multiple strains, including DOG01, DOG05, DOG06, CAT01, CAT02, and CAT03. The predicted major and minor parents of DOG05 came from Brazil. The DOG06 strain is potentially recombined from strains originating from humans and cats, whereas DOG01 is potentially recombined from G17 (ferret-originated) and Ave3 (chicken-originated), indicating that transmissions across species and regions may occur. Sixteen representative amino acid mutation sites were identified: nine in VP1 (12 R/H, 114S/N, 123I/M, 167 L/P, 231 P/S, 237 P/L, 243 R/W, 335 T/A, and 444S/N), four in VP2 (81 A/P, 103 R/H, 223 R/G, and 228 A/T), and three in VP3 (38 M/I, 61 A/T, and 65 V/A). These mutations were only harbored in strains identified in dogs and cats in this study. Whether this is related to host tropism needs further investigation. In this study, GyVg1 was identified in the sera of dogs and cats, and the molecular characteristics prompted the attention of public health.

Development and validation of the perceived symptom manageability scale among people living with the human immunodeficiency virus.

BACKGROUND: "Perceived Symptom Manageability (PSM)" is essential in symptom management among people living with HIV. As a standardized assessment instrument was lacking, we developed a PSM scale for people living with human immunodeficiency virus (PSM-HIV). METHODS: Data analysis was performed using the sample from HIV-designated medical institutions (N = 540). Psychometric testing, namely reliability and validity, is assessed by unidimensionality, internal consistency, exploratory and confirmatory factor analysis, and structural equation modeling. RESULTS: The final version of the PSM- HIV scale contained 15 items. This scale was submitted to a principal components analysis with varimax rotation, and three factors were obtained, explained by a total variance of 63.10%. The three factors were named Cognitive-Behavioral, Affective Interaction, and Self-Attitude. The results show that the scale had high reliability, Cronbach's α of the scale ranged from 0.71 to 0.92, and the Intraclass Correlation Coefficient was 0.88. The structural equation model supports a factor model with the acceptable fit (χ2/df (CMIN/DF) = 2.50, Root Mean square Residual (RMR) = 0.03, Goodness-of-Fit Index (GFI) = 0.93, Adjusted Goodness of Fit Index (AGFI) = 0.90, Normed Fit Index (NFI) = 0.93, Incremental Fit Index (IFI) = 0.96, Comparative Fit Index (CFI) = 0.96). The average variance extracted was 0.38 ∼ 0.59, and the composite reliability was 0.70 ∼ 0.91, indicating that the convergent validity of the scale is acceptable. Subjects with different stages of the disease reached significance(χ2 = 9.02; df = 2, P<0.05), meaning moderate Known-Groups Comparison Validation. CONCLUSIONS: The PSM-HIV scale is a valid instrument that measures overall attitude and belief about controlling or coping with HIV-relevant symptoms.

Molecular characterization of the Gyrovirus galga 1 strain detected in various zoo animals: the first report from China.

Since 2011, the Gyrovirus galga 1 (GyVg1, previously recognized as avian gyrovirus 2) strain has extensively been detected worldwide. The virus has been identified in several species, including chickens, humans, domestic cats, and snakes, especially in China. Therefore, in this study, the presence of GyVg1 was investigated in various zoo animals to determine whether it exists in various species in Nanyang, China. A total of 63 whole blood samples (1 sample from each animal) from 24 animal species were collected from the Nanyang Zoo. Eight different GyVg1 strains were identified in eight types of animals using polymerase chain reaction, and the full genome of each strain was sequenced. The whole genome of four GyVg1 strains, namely, HN2019-H1, HN2019-T1, HN2019-SD1, and HN2019-L1 identified in hippopotamus (Hippopotamus amphibius), tiger (Panthera tigris), sika deer (Cervus nippon), and lion (Panthera leo), respectively, comprised 2375 nucleotides (nt). The whole genome of the other strains, namely, HN2019-E1, HN2019-S1, HN2019-PF1, and HN2019-P1 identified in egret (Egretta garzetta), silver pheasant (Lophura nycthemera), peafowl (Pavonini), and common pheasants (Phasianus colchicus), respectively, comprised 2376 nt. Subsequently, a phylogenetic tree was constructed based on the 8 whole-genome sequence strains and 29 reference strains. These 37 strains were grouped into two major branches, group A and group B, and the 8 strains identified in this study were placed in group A. An analysis of the amino acids encoded by three open reading frames revealed some mutation sites unique to these eight strains. The substitution occurred at site 110 of viral protein 2 of HN2019-PF1, which is located in the highly conserved phosphatase motif WX7HX3CXCX5H (95-115aa). Recombination analysis revealed that, all these viral sequences were obtained as a result of recombination among the three GyVg1 strains (JL1511 and GS1512 from chickens and 17CC0810 from cat) from China and two strains (G17 from ferret of Hungary and RS-BR-15-2S from chicken of Brazil) from other countries. These findings indicate the complex evolution of GyVg1. Nevertheless, its transmission across the hosts is worth exploring.

First report of a novel goose astrovirus outbreak in Muscovy ducklings in China.

A highly acute disease characterized as visceral gout broke out in Muscovy ducklings in Henan province (China) in June 2020, with a mortality rate of up to 61%. In this study, common pathogenic agents were screened using reverse-transcription polymerase chain reaction or polymerase chain reaction. The results found the novel goose astrovirus (GoAstV) to be the pathogenic agent. We isolated the GoAstV, which has been designated as HNNY0620, using the Leghorn male chicken hepatocellular carcinoma (LMH) cell line and sequenced the complete genome. The phylogenetic tree showed that the amino acid (aa) sequences of ORF1a and ORF2 and the completed nucleotide sequences of the HNNY0620 strain were clustered in the GoAstV-I clade. ORF1a aa and whole-genome sequences were genetically close to TAstV-2 and DHV-3, whereas the ORF2 aa sequences were clustered with TAstV-2 and DHV2. Both the duck-origin GoAstVs and HNNY0620 harbored some special mutations, but ORF1a in 700 (I/T), ORF1b in 288 (F/L), and ORF2 in 306 (A/T) were only found in HNNY0620. These results suggest that the host range of GoAstV is diffusing, which can potentially affect other waterfowl.

Simple and Visible Detection of Novel Astroviruses Causing Fatal Gout in Goslings Using One-Step Reverse Transcription Polymerase Spiral Reaction Method.

In this study, a one-step isothermal method combining polymerase spiral reaction (PSR) with reverse transcription (RT-PSR) was established for rapid and specific detection of novel astroviruses causing fatal gout in goslings (N-GoAstV). The one-step RT-PSR was accomplished at the optimal temperature of 62°C and time of 40 min and used primers simply designed as conventional PCR primers, and the results of detection were visible to the naked eye. The detection limit of PSR was above 34.7 copies/µL at a 95% probability level according to probit regression analysis. The assay specifically detected N-GoAstV, and no other reference viruses were detected. These results suggest that the newly established RT-PSR assay could, in one step, accomplish reverse-transcription, amplification, and result determination providing a visible, convenient, rapid, and cost-effective test that can be carried out onsite, in order to ensure timely quarantine of N-GoAstV-infected birds, leading to effective disease control.

Novel genotype definition and genome characteristics of duck circovirus in central and Eastern China.

To explore genetic variations in duck circovirus (DuCV) and the molecular epidemiology of its infection, tissue samples were collected from 219 dead ducks from 20 farms in the central and eastern regions of China. All farms tested positive for DuCV, with duck-origin goose parvovirus, reovirus and Tembusu virus having co-infection rates of 100%, 0% and 0%, respectively. A total of 20 strains from the DuCV-positive flock were sequenced. The total sequence length was 1987-1996 nt, and the sequences shared 82% (JX499186, DuCV2 from Sichuan province, China) to 99.7% (KY328304, DuCV1 from Shandong Province, China) sequence identity with DuCV sequences available in GenBank. Hyper-variable regions were mainly located in open reading frame (ORF)2, ORF3 and intergenic regions. The tertiary structure of ORF2 from four provinces (Henan, Anhui, Zhejiang and Fujian) in China showed a canonical viral jelly roll and the antigenic epitope of ORF2 located in the bulge of the protein surface. Overall, 15 of the 20 DuCV strains are possibly derived through inter-genotypic and intragenotypic recombination. Based on sequence and phylogenetic analyses, six strains from Fujian Province clustered into a novel genotype-DuCV-1d. These findings may enrich our understanding of DuCV evolution and circulation and lay the foundation for vaccine strain selection.

Characterization and genomic analysis of emerging astroviruses causing fatal gout in goslings.

Since February 2017, severe outbreaks of fatal gout caused by novel gosling astroviruses (GoAstVs) have occurred in several Chinese provinces, causing a considerable economic impact on the poultry industry. To assess the infection status of GoAstVs causing gout, 165 clinical samples were collected from goslings from seven farms located in different Chinese provinces, and they were screened for viral infection. Seven GoAstV strains were completely sequenced. The positive infection rates of GoAstV, goose parvovirus, reovirus, goose haemorrhagic polyomavirus and Tembusu virus were 100%, 9.69%, 3.64%, 0% and 0%, respectively, indicating the role of GoAstV in gout. The genomes of all seven GoAstV strains were 7170-nt long and encoded three open reading frames (ORFs), namely, ORF1a, ORF1b and ORF2. Sequence and phylogenetic analyses of the seven GoAstV strains showed that these were avastroviruses and were closely related to viruses classified within Avastrovirus 3 and turkey astrovirus 2. Moreover, the mutation rates of ORF1a and ORF2 were high, and ORF1a was highly mutated at amino acid loci 545-580. The tertiary structure of the mutated ORF2 protein was smooth, and its antigenic epitope was highly mutated, which may be related to the pathogenicity of the virus and caused by antibody pressure from the host. These findings enrich our understanding of the evolution of novel GoAstVs causing gout and their circulation as well as lay the foundation for the selection of vaccine strains.

Rapid and visual detection of novel astroviruses causing fatal gout in goslings using one-step reverse transcription loop-mediated isothermal amplification.

To visually and rapidly detect a novel goose astrovirus (N-GoAstV) causing fatal gout in goslings, an isothermal detection method based on one-step reverse transcription loop-mediated isothermal amplification (one-step RT-LAMP) was established. The one-step RT-LAMP assay for N-GoAstV detection, using Bst 3.0 DNA polymerase with strong reverse transcription activity and primer sets targeting the opening reading frame 1b (ORF1b) of N-GoAstV, could be completed in 30 min using a water bath at 61°C; the detection results could be visually observed by adding a pH-sensitive dye containing phenol red and cresol red. The detection limit of the one-step RT-LAMP assay was 57.8 copies, which was similar to that of reverse transcription-quantitative polymerase chain reaction. The assay specifically detected N-GoAstV without any cross-reaction with other reference viruses, and this was further confirmed using enzyme digestion. These results indicated that the newly established RT-LAMP assay could accomplish reverse transcription, amplification, and visual result determination in one step, and the results obtained via this rapid and cost-effective method could be used to support disease control on farms in terms of N-GoAstV infection.