Summary of the Current Status of African Swine Fever Vaccine Development in China.

African swine fever (ASF) is a highly lethal and contagious disease of domestic pigs and wild boars. There is still no credible commercially available vaccine. The only existing one, issued in Vietnam, is actually used in limited quantities in limited areas, for large-scale clinical evaluation. ASF virus is a large complex virus, not inducing full neutralizing antibodies, with multiple genotypes and a lack of comprehensive research on virus infection and immunity. Since it was first reported in China in August 2018, ASF has spread rapidly across the country. To prevent, control, further purify and eradicate ASF, joint scientific and technological research on ASF vaccines has been carried out in China. In the past 4 years (2018-2022), several groups in China have been funded for the research and development of various types of ASF vaccines, achieving marked progress and reaching certain milestones. Here, we have provided a comprehensive and systematic summary of all of the relevant data regarding the current status of the development of ASF vaccines in China to provide a reference for further progress worldwide. At present, the further clinical application of the ASF vaccine still needs a lot of tests and research accumulation.

Opportunities and challenges for synthetic biology in the therapy of inflammatory bowel disease.

Inflammatory bowel disease (IBD) is a complex, chronic intestinal inflammatory disorder that primarily includes Crohn's disease (CD) and ulcerative colitis (UC). Although traditional antibiotics and immunosuppressants are known as the most effective and commonly used treatments, some limitations may be expected, such as limited efficacy in a small number of patients and gut flora disruption. A great many research studies have been done with respect to the etiology of IBD, while the composition of the gut microbiota is suggested as one of the most influential factors. Along with the development of synthetic biology and the continuing clarification of IBD etiology, broader prospects for novel approaches to IBD therapy could be obtained. This study presents an overview of the currently existing treatment options and possible therapeutic targets at the preclinical stage with respect to microbial synthesis technology in biological therapy. This study is highly correlated to the following topics: microbiota-derived metabolites, microRNAs, cell therapy, calreticulin, live biotherapeutic products (LBP), fecal microbiota transplantation (FMT), bacteriophages, engineered bacteria, and their functional secreted synthetic products for IBD medical implementation. Considering microorganisms as the main therapeutic component, as a result, the related clinical trial stability, effectiveness, and safety analysis may be the major challenges for upcoming research. This article strives to provide pharmaceutical researchers and developers with the most up-to-date information for adjuvant medicinal therapies based on synthetic biology.

Genome comparison of African swine fever virus China/2018/AnhuiXCGQ strain and related European p72 Genotype II strains.

African swine fever was introduced into China in August 2018 and led to high mortality in domestic pigs. We reported the genome characterization of the China/2018/AnhuiXCGQ strain mainly based on next-generation sequencing and comparison with related European p72 Genotype II strains. The genome was 189,393 bp long, encoding 181 open reading frames. Pair-wise genome sequence comparison revealed 54-107 variation sites between China/2018/AnhuiXCGQ and the other genotype II virulent strains contributing to the change of expression or alteration of amino acid residues in 10-38 genes. China/2018/AnhuiXCGQ strain shared the highest similarity with POL/2015/Podlaskie strain. Phylogenetic analysis based on a 125 kb long conserved central region revealed that the China/2018/AnhuiXCGQ strain and four European genotype II strains were grouped into three clusters. This study expanded our knowledge on the genetic diversity and evolution of ASFV and provided valuable information for diagnosis improvement and vaccine development.

Infection of African swine fever in wild boar, China, 2018.

On 16 November 2018, a wild boar infected with African swine fever was reported in China. The phylogenetic analysis showed that its causative strain belonged to the p72 genotype II, CD2v serogroup 8 and contained no additional tandem repeat sequences between the I73R and the I329L protein genes, which was different from previously reported strains in China.

Molecular Characterization of African Swine Fever Virus, China, 2018.

On August 3, 2018, an outbreak of African swine fever in pigs was reported in China. We subjected a virus from an African swine fever-positive pig sample to phylogenetic analysis. This analysis showed that the causative strain belonged to the p72 genotype II and CD2v serogroup 8.

Multiplex RT-PCR for rapid detection and differentiation of class I and class II Newcastle disease viruses.

A multiplex RT-PCR was developed for detection and differentiation of class I and class II strains of Newcastle disease virus (NDV). The method was shown to have high specificity and sensitivity. The results obtained from the multiplex RT-PCR for a total of 67 NDV field isolates obtained in 2009 were consistent with those obtained by nucleotide sequencing and phylogenetic analysis. A phylogenetic tree based on the partial sequences of the F gene revealed that the 67 field isolates of NDV could be divided into two classes. Twenty-seven NDV isolates were grouped into class I, and two genotypes were identified. Most of the class I isolates were determined to be of genotype 3, with the exception of isolate NDV09-034, which belonged to genotype 2. Forty class II NDV isolates were divided into three genotypes, namely genotype VII (27 isolates), genotype I (2 isolates) and genotype II (11 isolates). Isolates of genotypes I and II in class II were shown to be related to commercial vaccine strains used commonly in China. All isolates of genotype VII were predicted to be virulent, on the basis of the sequence motif at the cleavage site of the F gene. This genotype has become predominantly responsible for most outbreaks of ND in China in recent years. In conclusion, this multiplex RT-PCR provides a new assay for rapid detection and differentiation of both classes of NDV isolates.

Genomic characterization of the first class I Newcastle disease virus isolated from the mainland of China.

The complete genomic sequence of Newcastle disease virus (NDV) strain NDV08-004, isolated from domestic ducks in China, was determined in this study. The genome is 15198 nucleotides (nt) in length, follows the "rule of six" and contains a 55-nt leader sequence at the 3' end and a 114-nt trailer sequence at the 5' end. Compared with the full genome sequences of Class II NDV strains, the NDV08-004 isolate has a 12-nt insertion (TGGGAGACGGGG) in the phosphoprotein gene between nucleotides 2381 and 2382 of the genome (numbered according to the genomic sequence of the La Sota strain, which consists of 15186 nt). Strain NDV08-004 has the motif (112)E-Q-Q-E-R-L(117) at the cleavage site of the fusion protein, which is typical of lentogenic NDV strains, and this is in agreement with the results of pathogenic tests based on the mean death time (MDT) and the intracerebral pathogenicity index (ICPI). Phylogenetic analysis based on the full genome revealed that all the NDV strains studied could be divided into two distinct clades, namely class I and class II, and the NDV08-004 isolate characterized in this study was grouped in class I. Further phylogenetic analysis based on a 374-bp fragment of the F gene in class I strains of NDV demonstrated that NDV08-004 belongs to genotype 3, and should be therefore similar to strains obtained from live bird markets in Hong Kong in recent years.

Molecular characterization and phylogenetic analysis of new Newcastle disease virus isolates from the mainland of China.

Seventy-nine velogenic Newcastle disease virus (NDV) isolates were obtained from infected chicken flocks during the outbreaks of Newcastle disease (ND) in various regions of the mainland of China in 2006. The F gene fragment (535bp, from nt 47 to 581 of the F gene) which codes the main functional region of the F protein was obtained by RT-PCR and sequenced. All sequences obtained in this study have been submitted to GenBank. All the isolates have the motif (112)R-R-Q/R-K/R-R-F(117) at the cleavage site of the fusion protein, which is typical of velogenic NDV isolates. For genotyping, a phylogenetic tree based on nucleotides 47-435 of the F gene was constructed, and the 79 isolates could be divided into two genotypes, namely VIId and III. Most of the isolates proved to be of genotype VIId; only two isolates were of genotype III. Genotype VIId NDV has been the predominant pathogen responsible for most Newcastle disease outbreaks in China. The proportion of isolates of genotype VIId NDV shows an increasing trend, according to studies on the molecular epidemiology of NDV in China from 2002 to 2006.

Molecular epidemiological analysis of Newcastle disease virus isolated in China in 2005.

Eighty-three strains of Newcastle disease virus (NDV) were obtained from outbreaks in chickens, pigeons, geese, and ducks in China in 2005 and characterized genotypically. The main functional region of the F gene (535 nucleotides) was amplified and sequenced. A phylogenetic tree based on nucleotides 47-435 of the F gene was created using sequences from 83 isolates and representative NDV sequences obtained from GenBank. Phylogenetic analysis showed that all newly characterized strains belonged to six genetic groups: I, II, III, VIb, VIIc, and VIId. All the isolates belonging to groups I and II (14 total) were lentogenic according to the amino acid sequences of the fusion protein cleavage site, and either V4 or LaSota-type, depending on the vaccines that were used. Most isolates (64 total) were classified in group VIId, a predominant genotype responsible for most Newcastle disease outbreaks since the end of the last century. One strain, NDV05-055, was in group VIIc, three pigeon strains were in group VIb, and one isolate, NDV05-041, was in group III, and characterized as a velogenic strain. This study revealed that genotype VIId was the major NDV strain responsible for the 2005 ND epizoonosis that occurred in China.