Molecular epidemiological and genetic characterization of pseudorabies virus in Guangxi, China.

Pseudorabies virus (PRV) is an important pathogen that can cause harm to the pig population. Since 2011, there have been a number of large-scale outbreaks of pseudorabies on Chinese farms where animals had been vaccinated with the Bartha-K61 vaccine. In order to understand the epidemiological trend and genetic variations of PRV in Guangxi province, China, 819 tissue samples were collected from swine farms where PRV infection was suspected from 2013 to 2019, and these were tested for infectious wild strains of PRV. The results showed a positive rate of PRV in Guangxi province of 28.21% (231/819). Thirty-six wild-type PRV strains were successfully isolated from PRV-positive tissue samples, and a genetic evolutionary analysis was performed based on the gB, gC, gD, gE, and TK genes. Thirty of the PRV strains were found to be closely related to the Chinese variant strains HeN1-China-2012 and HLJ8-China-2013. In addition, five PRV strains were genetically related to Chinese classical strains, and one isolate was a recombinant of the PRV variant and the vaccine strain Bartha-K61. Amino acid sequence analysis showed that all 36 PRV strains had characteristic variant sites in the amino acid sequences of the gB, gC, gD, and gE proteins. Pathogenicity analysis showed that, compared to classical PRV strains, the PRV variant strains were more pathogenic in mice and had a lower LD50. Taken together, our results show that wild-type PRV infections are common on pig farms in Guangxi province of China and that the dominant prevalent strains were those of the PRV variants. The PRV variant strains also had increased pathogenicity in mice. Our data will provide a useful reference for understanding the prevalence and genetic evolution of PRV in China.

Isolation and identification of two novel pseudorabies viruses with natural recombination or TK gene deletion in China.

Pseudorabies virus (PRV), the causative agent of Aujeszky's disease, has gained increased attention in China in recent years due to outbreaks of emergent pseudorabies. However, there is limited information about the evolution and pathogenicity of emergent PRV field strains in China. In this study, two PRV field strains were isolated from an intensive pig farm with suspected PRV infection. These were named the GXLB-2015 and GXGG-2016 strains and their growth characteristics together with their genome sequences and pathogenicity were determined. Nucleotide homology and phylogenetic analysis revealed the GXLB-2015 stain was relatively close to the foreign PRV isolated strains with respect to the whole genome sequence. However, it formed an independent branch between the foreign PRV isolates and the previous PRV variants isolated in China. Further recombination and genetic evolution analysis showed that the GXLB-2015 strain was a natural recombinant between the Bartha strain and PRV variants. The GXGG-2016 strain was highly homologous with the Chinese classical strains, but it has a natural deletion of 69 aa in the thymidine kinase (TK) gene. Pathogenicity analysis showed that, the GXLB-2015 strain had the strongest pathogenicity to mice with an LD50 of 103.5, while the GXGG-2016 strain with the TK gene deletion was not pathogenic to mice. Taken together, our data provide direct evidence for the genomic recombination and natural TK gene deletion of PRVs, which may provide a reference for a better understanding of PRV evolution in China and contribute to the clinical control of PRV infection in pig farms.

Prophylactic Efficacy of Equine Immunoglobulin F(ab')2 Fragments Against Feline Parvovirus.

Feline parvovirus (FPV), a type of parvovirus prevalent worldwide, can cause foetal death and acute enteritis in adult cats with severe leukopenia, and yet there are no effective drugs to prevent or treat FPV. Here, the immune effects of two FPV vaccines on horses were compared. IgG was extracted from FPV-immunized horse sera. Equine F(ab')2 fragments were obtained from pepsin-digested IgG and then purified by protein-G column chromatography. The results showed that the inactivated FPV oil vaccine was more effective than the inactivated FPV propolis vaccine in helping healthy horses to produce hyper-immune serum. Four methods were tested, among which the optimized octanoic acid-ammonium sulphate precipitation method was proved to be the best process for extracting IgG. The optimal condition for preparing F(ab')2 by pepsin digestion was 30 °C for 3.5 h, and the content, purity and recovery of F(ab')2 were 8.64 mg/mL, 90.36% and 93.24%, respectively. Our equine immunoglobulin F(ab')2 fragments effectively neutralized activity in vitro against FPV, alleviated the clinical symptoms of FPV-infected cats, reduced the viral loads in the intestine and had prophylactic effects in FPV-infected cats. These results indicate that the F(ab')2 fragment prepared from inactivated FPV-immunized horses may be used as a prophylactic agent for diseases caused by FPV.

Isolation, genetic analysis of the first Akabane virus from goat in China.

Akabane virus (AKAV) is an important insect-borne virus belonging to the genus Orthobunyavirus, the Peribunyaviridae family. An AKAV defined as GXDH 01 here, was isolated for the first time from blood from a sentinel goat in China in 2016, and its full-length open reading frames (ORFs) were sequenced in this study. Sequence analysis suggested that the isolate GXDH 01 probably had undergone a reassortment incident and acquired L segments from other strain originating from an attenuated vaccine, such as OBE-1. This study aims to provide more understanding as to the origin and epidemiology of AKAV in China.

[Geographical variation of Glycyrrhiza uralensis seed germination character on water stress].

OBJECTIVE: We studied the licorice provenance rule under the manual simulative water stress conditions, discussed geographical variation pattern and the ecology mechanism and laid the theoretical basis for the licorice seed regionalization and the seed allocation. METHOD: On the spot we investigated and collected seed materials. Setting up water potential gradient by PEG, we observed the seed germination characteristic through the experiment of indoor germination. RESULT: The sensitivity of various provenance seed germinative energy show quite remarkable difference for water potential reducing. Along with geographic longitude increasing, the seed germinates variation sensitivity increasing gradually to the water stress. The results of climatic factors' correlation analysis indicated that provenance seed germinate energy to the water stress is sensitive, in which the annual average temperature is low and annual precipitation is abundant. But it is insensitive in which the annual precipitation is little, the annual average temperature, average temperature in July and the annual average ground temperature is high. CONCLUSION: Under the water stress condition the licorice provenance seed germination characteristic exists remarkable geographical variation. Geographical variation is the result of natural selection.

[Study on geographical variation of morphologic and germination characteristic of different Glycyrrhiza uralensis provenance seeds].

OBJECTIVE: To study the geographical variation of morphologic and germination characteristic of different Glycyrrhiza uralensis provenance seeds, approach the geographical variation mode and ecology mechanism, and laid theoretical foundation for districting and allocating of G. uralensis seeds. METHOD: Field investigation and laboratory analysis were applied. Seed shape and kilosseed weight were sampled randomly, germination rate germination force by general methods. RESULT: The morphologic characteristic of G. uralensis seeds showed roughly longitude variation tendency that the seeds increased gradually from west to east. While the germination characteristic showed roughly altitude variation tendency that the seeds germination rate and germination force increased with the increase of the altitude, and the average germination rate was the same with the seeds morphologic characteristic. The results of analysis correlated with the climatic factors show that the morphologic characteristic of G. uralensis was positive correlated with annual rain-fall of the habitat, and the germination rate was quickened by drought, high temperature and strong sunshine. CONCLUSION: The morphologic and germination characteristic and of G. uralensis seeds present distinguished geographical variation, and the formation of the variation was related to the ecological environment in which the seed provenance adapted.

[Studies on the shade-endurance capacity of Glycyrrhiza uralensis].

OBJECTIVE: To study the shade-endurance property of Glycyrrhiza uralensis and provide rationale for the practice of inter-cropping G. uralensis with trees. METHOD: Black shading nets were used to provide five different environments of light intensities (light penetration rates of 100%, 75%, 65%, 50% and 25%, respectively). To assess the shade-endurance capacity of G. uralensis, several aspects were evaluated, including growth characters, physiological and ecological characters, biomass, and chemical contents. RESULT AND CONCLUSION: G. uralensis is a light-favored plant. The growth indices such as plant height, stem diameter, leaves number, root diameter, biomass, and daily average photosynthetic rate (Pn) are highest when light permeation rate is 100%. All these indices decrease when light intensity decreases. However, G. uralensis possesses shade-endurance capacity to some degree; it adapts to the shading environment by increasing the leaf area and chlorophyll contents. Shading has no obvious effect on the absolute light energy utilization rate (Eu) or Fv/Fm ratio. The influence of shading on the chemical contents of G. uralensis is obvious.