Publications by authors named "Yingzuo Bi"

54 Publications

Epidemiological investigations and locally determined genotype diversity of Mycoplasma synoviae in Central China from 2017 to 2019.

Poult Sci 2021 Oct 10;101(1):101522. Epub 2021 Oct 10.

College of Animal Science, South China Agricultural University, Guangzhou 510642, PR China.

Mycoplasma synoviae (M. synoviae) has been identified worldwide to cause respiratory diseases, infectious synovitis, airsacculitis, and eggshell apex abnormalities (EAA) in commercial chickens, which results in substantial economic losses to the poultry industry. Therefore, in this study, 258 flocks were investigated between 2017 and 2019 for M. synoviae by screening samples from Central China. Subsequently, 129 M. synoviae strains were isolated, with a positive rate of 50%. Moreover, a higher incidence of M. Synoviae infections was in layers (74.1%) than in broilers (20%) in this study. The 5'-end conserved segment of the variable lipoprotein hemagglutinin A (vlhA) gene of these isolates was then cloned and sequenced because it is a common genomic target identified so far for M. synoviae genotyping. Genotyping of all isolates was based on the phylogenetic analysis and length analysis of the proline-rich-repeat (PRR) regions, respectively. Phylogenetic analysis based on 5'-end conserved segment of the vlhA gene (76-421 nt) assigned the majority of the occurring strains as being from group 6, and others from groups 2 and 3. Results identified that these isolates were of 6 types: A (38aa), D (23aa), E (19aa), I (28aa), J (20aa), and L (35aa), based on the size of the PRR region analysis. Furthermore, most of the isolates (81.4% were identified as type L. Additionally, the epidemic types included only I and L in 2017; however, the types rose to 5 (A, D, E, I, L) in 2018 and rose to 6 (A, D, E, I, J, L) in 2019. These data showed the genotype diversity of M. synoviae in Central China. The high rate of positive flocks suggests the urgent need to take real-time supervisory controls of this Mycoplasma species in avian flocks.
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http://dx.doi.org/10.1016/j.psj.2021.101522DOI Listing
October 2021

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

Poult Sci 2021 Oct 26;100(10):101407. Epub 2021 Jul 26.

College of Animal Science, South China Agricultural University, Guangzhou 510642, PR 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.
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http://dx.doi.org/10.1016/j.psj.2021.101407DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8383103PMC
October 2021

Genetic Characterisation and Local Genotypes of Canine Parvovirus Strains Collected from Pet Dogs in Central and Eastern China During 2018-2019.

J Vet Res 2020 Dec 19;64(4):477-486. Epub 2020 Nov 19.

College of Animal Science, South China Agricultural University, Guangzhou 510642, PR China.

Introduction: Canine parvovirus type-2 (CPV-2) causes acute infectious diseases in puppies, which show high morbidity and mortality. Better effect of vaccination against these diseases could be achieved with deeper knowledge of CPV-2 genotype dissemination and mutation history. This study investigated CPV-2-positive samples collected recently over a wide region of China.

Material And Methods: A total of 118 faecal samples from dogs identified as CPV-positive were collected from veterinary clinics in central and eastern China. Overall, 16 strains collected from Anhui, 29 from Henan, and 16 from Zhejiang Province were sequenced to determine the genotypic composition of CPV-2 and mutational complexity of CPV-VP2.

Results: The CPV-2a, CPV-2b, and CPV-2c genotypes were detected in Anhui and Henan Provinces, while CPV-2c alone was detected in Zhejiang Province. Sequence analysis of all strains showed 98.5%-99.8%, 98.3%-99.9%, and 98.7%-99.8% identity among the 16 Anhui, 29 Henan, and 16 Zhejiang strains, respectively. Strains collected from Anhui and Henan Provinces showed lower identity (97.0%), suggesting greater genetic divergence in central China. The mutation rates of Henan and Anhui strains were lower than that of Zhejiang strains. Major amino acid mutations occurred at sites 5, 370, 426, and 440. Epitope and entropy analyses implied these sites' likely conformance to the principles of mutation tendency, complexity, and diversity.

Conclusion: The findings for the evolutionary structure of CPV-2 strains collected from three provinces in central and eastern China advance trend monitoring of the genetic variation in canine parvovirus and point to its implications in the development of novel vaccines.
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http://dx.doi.org/10.2478/jvetres-2020-0076DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7734690PMC
December 2020

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

Front Vet Sci 2020 10;7:579432. Epub 2020 Dec 10.

College of Animal Science, South China Agricultural University, Guangzhou, China.

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.
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http://dx.doi.org/10.3389/fvets.2020.579432DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7758545PMC
December 2020

An Improved Polymerase Cross-Linking Spiral Reaction Assay for Rapid Diagnostic of Canine Parvovirus 2 Infection.

Front Vet Sci 2020 30;7:571629. Epub 2020 Oct 30.

College of Animal Science, South China Agricultural University, Guangzhou, China.

With increasing complications of canine parvovirus infection cases, disease diagnosis and treatment have become more difficult. In this study, specificity primers for the conserved region of the VP2 gene of canine parvovirus 2 (CPV-2) were synthesized and evaluated. An improved polymerase cross-linking spiral reaction (PCLSR) method for early and rapid diagnosis of CPV-2 was established. The results showed that the amplification reaction was optimal when run at 62°C for 50 min and could be used to detect CPV-2 without any cross-reactions with other pathogens of canine infectious diseases. Reaction results were directly judged by the naked eyes, with the positive amplification tube shown as luminous yellow and the negative tube as bright purple. Compared with the previously reported polymerase spiral reaction (PSR) method for CPV-2 detection, this reaction was performed using improved primer pairs and a better dye identification method (using an indicator comprising phenol red and cresol red). The detection limit of PCLSR was 3.9 × 10 copies using gel electrophoresis or a visible dye. The positive rate of 132 clinical samples was 42.42%, which was identically the same as that of the PSR method and slightly higher than that of the colloidal gold strip method (39.39%). The newly developed CPV-PCLSR assay shows the advantage of rapid visualization of results and offers a convenient and rapid method for early CPV-2 diagnosis with higher sensitivity and specificity than the established methods.
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http://dx.doi.org/10.3389/fvets.2020.571629DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7661784PMC
October 2020

Molecular Characterisation and Genetic Diversity of Canine Parvovirus Type 2 Prevalent in Central China.

J Vet Res 2020 Sep 16;64(3):347-354. Epub 2020 Sep 16.

College of Animal Science, South China Agricultural University, Guangzhou, 510642, PR China.

Introduction: Canine parvovirus (CPV) disease is one of the most threatening to domestic and wild dogs.

Material And Methods: A total of 132 clinical samples were isolated from domestic dogs with diarrhoea from Henan, Hubei, Jiangsu, and Anhui provinces from 2016 to 2017, and 56 were positive for CPV-2 by PCR. A phylogenetic tree was constructed for the isolate sequences incorporating 53 non-Chinese reference strains.

Results: VP2 sequences showed the strains mainly to be new CPV-2a/2b and CPV-2c genotypes. The Ala5Gly, Phe267Tyr, Ser297Ala, Tyr324Ile, Gln370Arg, Asn426Asp or Asn426Glu, and Thr440Ala sites in the VP2 protein antigenic region were found to have high mutation rates. The VP2 tertiary structural model shows that the change at these mutation points is a factor for the changes in the protein structure. Significant differences between the Central Chinese strains and others were found, indicating that evolution is geographically related and extended in major regions. The homology between the identified strains confirmed their relationship. Phylogenetic analysis indicated that the common genotypes in the same clusters differ slightly in homology and evolutionary history.

Conclusion: This epidemiological study enriches the available data and serves as an important reference for studies on the evolution of CPV and selection of vaccines in China.
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http://dx.doi.org/10.2478/jvetres-2020-0056DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7497761PMC
September 2020

Novel Genotype Definition and the First Epidemiological Investigation of Canine Adenovirus Type 2 in Dogs in Central China.

Front Vet Sci 2020 19;7:534. Epub 2020 Aug 19.

College of Animal Science, South China Agricultural University, Guangzhou, China.

Infections caused by canine adenovirus (CAdV) type 1 have been reported worldwide in the past two decades. However, only few studies have specifically reported the prevalence of CAdV type 2 (CAdV-2). The present study investigated the persistent circulation of CAdV-2 in dogs with diarrhea in the Henan, Hubei, and Jiangsu provinces in central China from 2017 to 2019. We conducted polymerase chain reaction for detecting CAdV-2 and other related pathogens in 224 rectal swabs of pet dogs and the co-infection of canine diseases was also analyzed. In addition, the structural protein genes-Fiber, Hexon, and Penton-of the isolated CAdV-2 strains were sequenced and analyzed. The similarity between and among the 19 strains was 97.4%, as revealed by sequence alignment. Multiple sequence alignment results showed that the gene sequences of these CAdV-2 strains shared 97.4-99.8% nucleotide and 94.1-99.3% amino acid identity with reference sequences and shared only 79.0-80.5% nucleotide and 77.3-80.5% amino acid identity with the vaccine strain CLL, indicating that Fiber harbored most of the variant sites. Furthermore, pairwise sequence comparisons of of CH-JS-1901 and CH-HN-1801 with that of India2006 revealed a novel genotype. Furthermore, protein model prediction showed that the amino acid mutation of fiber protein in 19 strains was located in the head region, that may cause structural changes on the surface of the fiber protein. These findings are of significance for monitoring the epidemiology of CAdV-2 infection and developing a novel vaccine which contribute to understanding genetic evolution of CAdV-2 in China.
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http://dx.doi.org/10.3389/fvets.2020.00534DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7466760PMC
August 2020

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

Transbound Emerg Dis 2020 Nov 24;67(6):2993-3004. Epub 2020 Jun 24.

College of Animal Science, South China Agricultural University, Guangzhou, PR 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.
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http://dx.doi.org/10.1111/tbed.13676DOI Listing
November 2020

Epidemiological investigation of avian infectious bronchitis and locally determined genotype diversity in central China: a 2016-2018 study.

Poult Sci 2020 Jun 10;99(6):3001-3008. Epub 2020 Apr 10.

College of Animal Science, South China Agricultural University, Guangzhou 510642, P.R. China.

Infectious bronchitis (IB), caused by avian IB virus (IBV), is an acute and highly contagious disease of chickens. From 2016 to 2018, 56 IBV strains were isolated and identified from clinical samples obtained from various chicken farms located in central China. The S1 sequencing of these strains revealed nucleotide and amino acid identities of 70.2 to 100% and 62.6 to 100%, respectively, compared with those of reference strains. Phylogenetic analysis indicated that the genotypes of the isolates included GI-13 (4/91), GI-7 (TW-I), GI-24 (Mass), GI-19 (QX), and GI-18 (LDT3-A), with GI-19 (QX) being the predominant genotype. Meanwhile, GI-13 (4/91) was the second most dominant genotype in Henan Province, whereas it was GI-7 (TW-I) in Hunan and Hubei provinces. Recombination analysis of 3 variant strains showed that CK/CH/HeN/20160113 might be a recombination of LDT3-A- and QX-type strains and that CK/CH/HeN/20160316 might be a recombination of Italy-02-type strain and CK-CH-LJS08II. The predicted tertiary structure between CK/CH/HeN/20160113 and LDT3-A-type strain revealed that the novel 336 (L-P) and 455 (S-A) mutations changed the structure from an alpha helix to a random crimp. In addition, the 275 (Y-F) site reduced the length of the β-sheet, whereas the site 353 (A-T) extended the β-sheet. These findings suggested that GI-19 (QX) remains the predominant genotype in central China, and a locally determined complex genotype associated with variable clinical symptoms exists related to gene recombination and mutations.
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http://dx.doi.org/10.1016/j.psj.2020.03.023DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7597734PMC
June 2020

Simple and visible detection of duck hepatitis B virus in ducks and geese using loop-mediated isothermal amplification.

Poult Sci 2020 Feb 24;99(2):791-796. Epub 2020 Jan 24.

College of Animal Science, South China Agricultural University, Guangzhou 510642, People's Republic of China.

In this study, loop-mediated isothermal amplification (LAMP) was used to establish a rapid, specific, and visual detection method for duck hepatitis B virus (DHBV). The design and synthesis of 4 specific LAMP primers were based on the conserved gene region of the DHBV genome, and the optimum temperature and time of the LAMP reaction were 63°C and 50 min, respectively. The LAMP assay was confirmed to be specific for DHBV detection and had the same sensitivity as the quantitative PCR assay. A visual detection method for rapid determination of results was developed using a color indicator containing phenol red and cresol red. A color change was produced based on a pH change in the reaction system, indicating a positive reaction. For the detection of samples from ducks and geese, the LAMP method has the advantages of simplicity, high sensitivity and specificity, good visibility, and low cost. Moreover, it is more practical and convenient than PCR-related assays for the clinical detection of DHBV.
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http://dx.doi.org/10.1016/j.psj.2019.12.024DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7587725PMC
February 2020

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

Transbound Emerg Dis 2020 Mar 17;67(2):865-876. Epub 2019 Nov 17.

College of Animal Science, South China Agricultural University, Guangzhou, China.

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.
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http://dx.doi.org/10.1111/tbed.13410DOI Listing
March 2020

Novel polymerase spiral reaction assay for the visible molecular detection of porcine circovirus type 3.

BMC Vet Res 2019 Sep 6;15(1):322. Epub 2019 Sep 6.

College of Animal Science, South China Agricultural University, Guangzhou, 510642, People's Republic of China.

Background: Porcine circovirus type 3 (PCV3) is a newly emerging circovirus that might be associated with porcine dermatitis and nephropathy syndrome, reproductive failure, and cardiac and multisystemic inflammation. To aid the prevention and control of the infectious disease caused by PCV3, we developed a novel isothermal amplification assay using polymerase spiral reaction (PSR), which allows the visual detection of preserved strains and clinical samples.

Results: This assay precisely amplified the PCV3 genome with the use of a water bath at 62 °C for 50 min. The detection limit was found to be 1.13 × 10 copies/μL by gel electrophoresis or with the use of a visible dye (an indicator comprising phenol red and cresol red). No cross-reaction with other porcine infectious viruses was observed. The detection results for 23 PCV3-positive samples by PSR were in accordance with loop-mediated isothermal amplification (LAMP) assay. The detection rate of the PSR assay for PCV3 positivity of clinical samples was 68/97, which was higher than LAMP assay (67/97).

Conclusions: These results indicated that the PSR assay provides an accurate and rapid method for the detection of PCV3 with high sensitivity and specificity. It is particularly suited for use in a simple laboratory setting without a thermal cycler or gel electrophoresis equipment.
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http://dx.doi.org/10.1186/s12917-019-2072-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6731610PMC
September 2019

Rapid and visible detection of Mycoplasma synoviae using a novel polymerase spiral reaction assay.

Poult Sci 2019 Nov;98(11):5355-5360

College of Animal Science, South China Agricultural University, Guangzhou 510642, PR China.

In this study, a rapid, specific, and sensitive detection assay for Mycoplasma synoviae (MS) was established using a polymerase spiral reaction (PSR) method. A pair of primers were designed according to the conserved region of the vlhA gene of MS, and PSR results were assessed using agarose gel electrophoresis and color rendering with a dye indicator. The optimum reaction temperature and time for PSR using the specific primers were 62°C and 40 min in a water bath, respectively. The sensitivity of the PSR assay for MS detection was 100 times more than that of the polymerase chain reaction assay based on agarose gel electrophoresis results and color change detected by the naked eye. Further experiments demonstrated that the primers specifically detected MS and showed no cross-reaction with other prevalent avian pathogens. Clinical sample testing confirmed that the MS-PSR assay is simple, rapid, specific, and sensitive, and thereby very suitable for application and promotion in the field and laboratories of grassroots units.
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http://dx.doi.org/10.3382/ps/pez356DOI Listing
November 2019

Visual detection of porcine epidemic diarrhea virus using a novel reverse transcription polymerase spiral reaction method.

BMC Vet Res 2019 Apr 15;15(1):116. Epub 2019 Apr 15.

College of Animal Science, South China Agricultural University, Guangzhou, 510642, People's Republic of China.

Background: Porcine epidemic diarrhea virus (PEDV) is a major etiological agent of porcine epidemic diarrhea around the world. Point-of-care testing in the field is lacking owing to the requirement for a simple, robust field applicable test that does not require professional laboratory equipment. The aim of this study was to establish a novel reverse transcription polymerase spiral reaction (RT-PSR) assay for the rapid detection of porcine epidemic diarrhea virus (PEDV). For the assay, a specific RT-PSR primer pair was designed against a conserved region in PEDV ORF3.

Results: The RT-PSR was optimized, and PEDV could be detected after a 50 min incubation at 62 °C, in addition to the 15 min required for reverse transcription. No cross-reaction with other porcine infectious viruses was observed. This new method for PEDV detection was 10 times more sensitive than the conventional reverse transcription-polymerase chain reaction (RT-PCR) assay. The positive rates for 65 clinical samples using the new RT-PSR assay and the conventional RT-PCR assay were 58.46% (38/65) and 53.84% (35/65), respectively. In the RT-PSR assay, the addition of a mixture of dyes allowed a positive reaction to be directly observed by the naked eye.

Conclusions: These results indicate that this RT-PSR assay is capable of accurately detecting PEDV, and has the advantages of high specificity and sensitivity for the detection of PEDV.
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http://dx.doi.org/10.1186/s12917-019-1851-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6466714PMC
April 2019

Detection of three novel atypical porcine pestivirus strains in newborn piglets with congenital tremor in southern China.

Infect Genet Evol 2019 03 6;68:54-57. Epub 2018 Dec 6.

College of Animal Science, South China Agricultural University & Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, Guangzhou 510642, PR China. Electronic address:

Atypical porcine pestivirus (APPV) have been discovered in swine herds from three provinces in China, suggesting a wide distribution in China. This study reports the occurrence of three novel APPV strains in China. They were detected from newborn piglets with clinical signs of congenital tremors (CT) in Guangdong Province, China. The complete genomic sequences of three novel APPV strains exhibited only 80.5%-84.1% nucleotide sequences homology with other APPV reference sequences available in GenBank. Phylogenetic analysis showed that these novel APPV strains formed independent branch from the American, German, Netherlandish, Australian and other Chinese strains. These results will help us better understand the epidemiology and genetic characteristics of APPV in China.
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http://dx.doi.org/10.1016/j.meegid.2018.12.008DOI Listing
March 2019

Expression of dysregulated miRNA in vivo in DF-1 cells during the course of subgroup J avian leukosis virus infection.

Microb Pathog 2019 Jan 23;126:40-44. Epub 2018 Oct 23.

Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding and Key Lab of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, Guangzhou, 510642, PR China; Key Laboratory of Animal Health Aquaculture and Environmental Control, Guangdong Provincial Department of Science and Technology, Guangzhou, 510642, PR China; College of Animal Science, South China Agricultural University, Guangzhou, 510642, PR China. Electronic address:

Aberrant expression of microRNAs (miRNAs) is known to be involved in cancer progression caused by subgroup J avian leukosis virus (ALV-J) in liver tissues. To advance our understanding of the related pathological mechanisms and virus-host interactions, seven previously reported miRNAs were selected for a comparative analysis of miRNA expression between infected and uninfected DF-1 cells, including six miRNAs related to tumorigenesis (let-7b/7i, miR-221/222, miR-125b, miR-375 and miR-2127. The results showed that six of the seven miRNAs except gga-miR-375 were upregulated in cells infected with NX0101 (caused myeloma (ML)) and GD1109 (caused hemangioma (HE)) at 1 h post infection. On day 2 post-infection, all seven miRNAs were upregulated in infected DF-1 cells. On day 6 post-infection, gga-let-7b, gga-miR-125b, and gga-miR-375 were downregulated whereas gga-miR-221 and gga-miR-222 were upregulated in DF-1 cells infected with the two ALV-J strains of different phenotypes. However, expression of gga-let-7i was reduced in DF-1 cells infected with NX0101 and was increased in those infected with GD1109; gga-miR-2127 expression showed no significant difference between infected and uninfected cells. This study is the first to report the changes in the miRNA expression levels in DF-1 cells during the course of ALV-J infection, and suggests a relationship between its pathological mechanisms and miRNAs.
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http://dx.doi.org/10.1016/j.micpath.2018.10.027DOI Listing
January 2019

Epidemiology and characterization of avian infectious bronchitis virus strains circulating in southern China during the period from 2013-2015.

Sci Rep 2017 07 26;7(1):6576. Epub 2017 Jul 26.

College of Animal Science, South China Agricultural University, Guangzhou, 510642, P.R. China.

Two hundred and six strains of avian infectious bronchitis virus (IBV) were isolated from chickens showing signs of disease in southern China during the period from 2013-2015. The nucleotide and amino acid sequences from the isolated field strains were compared to 42 published references. Nucleotide homologies ranged from 63.1-99.9% and amino acid homologies ranging from 60.2-100%. At least seven IBV genotypes were co-circulating in commercial chicken farms in southern China. The IBV isolates were genetically diverse and underwent continuing evolution. The QX-type, TW I-type, and 4/91-type were the most common genotypes during the three-year observation period and accounted for 88.8% of the isolated strains. Notably, the prevalence of the TW I-type strains has been increasing in recent years and has become the most common genotype in China. The emergence of variant IBV strains can be attributed to recombination. Serologic analysis and antigenic 3D cartography of 4 reference and 14 field isolated strains indicated the surveyed IBVs had diverse serology types and that the serotype of the isolated QX-type and TW I-type strains was distinct from the vaccines strains. Therefore, long-term continuing surveillance is necessary for IBV prevention and control.
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http://dx.doi.org/10.1038/s41598-017-06987-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5529424PMC
July 2017

Temporal changes of microRNA gga-let-7b and gga-let-7i expression in chickens challenged with subgroup J avian leukosis virus.

Vet Res Commun 2017 Sep 11;41(3):219-226. Epub 2017 Feb 11.

Breeding and Reproduction, Ministry of Agriculture, Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding and Key Lab of Chicken Genetics, Guangzhou, 510642, People's Republic of China.

Two important microRNAs, gga-let-7b and gga-let-7i were examined for the relative expression in liver and bone marrow tissues from specific pathogen free chickens that were challenged either with GD1109 or NX0101 strain of subgroup J avian leukosis virus (ALV-J). The GD1109 strain of ALV-J reportedly causes hemangioma (HE) and NX0101 reportedly causes myeloma (ML) in susceptible chickens. Temporal changes of both gga-let-7b and gga-let-7i expression in ALV-J infected chickens were observed in contrast to its counterpart of a non-infected negative control group of chickens (P < 0.05 or P < 0.01) during the first 120 days post infection. Use of the web-based computational DIANA-mirPath software (available at http://microrna.gr/mirpath ), it was predicted that both gga-let-7b and gga-let-7i were involved in multiple pathways including signaling pathways, such as MAPK, TGF-beta, Notch, Wnt, mTOR, Cell cycle, P53 and Jak-STAT. Combining our experimental data with reports on the microRNAs, we suggest that both gga-let-7i and gga-let-7b may also act as tumor suppressors in chicken, especially play a critical role in tumorigenesis induced by ALV-J.
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http://dx.doi.org/10.1007/s11259-017-9681-1DOI Listing
September 2017

Characterization of field strains of infectious laryngotracheitis virus in China by restriction fragment length polymorphism and sequence analysis.

J Vet Diagn Invest 2016 Jan 23;28(1):46-9. Epub 2015 Dec 23.

College of Animal Science, South China Agricultural University, Guangzhou, China (Yan, Li, Xie, Chen, Bi)Key Laboratory of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, Guangzhou, China (Xie, Chen).

Nineteen strains of infectious laryngotracheitis virus (ILTV; Gallid herpesvirus 1) were isolated from dead or diseased birds in chicken flocks from different areas of China between 2010 and 2014 and used to investigate ILTV epidemiology. These strains were characterized using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) patterns and sequence analysis of the thymidine kinase (TK) gene. PCR-RFLP analysis showed that the TK gene generated 2 patterns when digested with restriction endonuclease enzymes. Pattern A corresponded to 2 virulent field strains, while pattern B was characteristic of 2 virulent field strains, 15 low pathogenicity field strains, and all vaccine strains. Sequence analysis of the TK gene indicated that the messenger RNA polyadenylation signals could be identified in some isolates where amino acid 252 was threonine, and in those with methionine at that position. The present study has demonstrated that most of the outbreaks of ILT in China were caused either by low virulence strains or by vaccine-related strains, and also emphasizes the importance of reinforcing ILTV surveillance in both vaccinated and nonvaccinated flocks.
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http://dx.doi.org/10.1177/1040638715618230DOI Listing
January 2016

Role of gga-miR-221 and gga-miR-222 during Tumour Formation in Chickens Infected by Subgroup J Avian Leukosis Virus.

Viruses 2015 Dec 11;7(12):6538-51. Epub 2015 Dec 11.

College of Animal Science, South China Agricultural University & Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, Guangzhou 510642, China.

Subgroup J avian leukosis virus (ALV-J) causes a neoplastic disease in infected chickens. Differential expression patterns of microRNAs (miRNAs) are closely related to the formation and growth of tumors. (1) BACKGROUND: This study was undertaken to understand how miRNAs might be related to tumor growth during ALV-J infection. We chose to characterize the effects of miR-221 and miR-222 on cell proliferation, migration, and apoptosis based on previous microarray data. (2) METHODS: In vivo, the expression levels of miR-221 and miR-222 were significantly increased in the liver of ALV-J infected chickens (p < 0.01). Over-expression of gga-miR-221 and gga-miR-222 promoted the proliferation, migration, and growth of DF-1 cells, and decreased the expression of BCL-2 modifying factor (BMF) making cells more resistant to apoptosis. (3) RESULTS: Our results suggest that gga-miR-221 and gga-miR-222 may be tumour formation relevant gene in chicken that promote proliferation, migration, and growth of cancer cells, and inhibit apoptosis. BMF expression was significantly reduced in vivo 70 days after ALV-J infection. They may also play a pivotal role in tumorigenesis during ALV-J infection.
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http://dx.doi.org/10.3390/v7122956DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4690879PMC
December 2015

Assessing the efficacy of an inactivated chicken anemia virus vaccine.

Vaccine 2015 Apr 8;33(16):1916-22. Epub 2015 Mar 8.

College of Animal Science, South China Agricultural University, Guangzhou 510642, PR China; Key Laboratory of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, Guangzhou 510642, PR China; Key Laboratory of Animal Health Aquaculture and Environmental Control, Guangzhou 510642, PR China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510640, PR China. Electronic address:

Background: Chicken anemia virus (CAV) is an immunosuppressive virus that causes chicken infectious anemia (CIA) which is a highly contagious avian disease. CAV causes major economic losses in the poultry industry worldwide. The current CAV vaccine is a live attenuated strain administered in the drinking water that risks horizontal infection of other chickens. The purpose of this study was to develop a novel vaccine against CAV that can be administered safely using a highly pathogenic isolate inactivated with β-propiolactone hydrolysis that would protect chicks from CAV.

Methods: Hens were vaccinated twice intramuscularly with a novel CAV GD-G-12 inactivated vaccine and the humoral immune responses of the hens and offspring were monitored by ELISA. A heterologous intramuscular challenge using the CAV strain GD-E-12 was conducted in the chicks hatched from vaccinated or unvaccinated hens.

Results: The vaccine strain, GD-G-12, was shown to be highly pathogenic prior to inactivation evidenced by thymic atrophy and bleeding, and weight loss. The inactivated vaccine was considered safe and showed no signs of pathogenicity. High titers of CAV specific antibodies were detected in the vaccinated hens and in their chicks, indicating vertical transfer of maternal antibodies. Furthermore, the chicks hatched from vaccinated hens were resistant to a heterologous CAV challenge and showed no signs of weight loss and thymic atrophy and bleeding.

Conclusion: Our studies are proof of principle that inactivated GD-G-12 might be a novel vaccine candidate to prevent CAV infection, and highlight the utility of using an inactivated virus for this vaccine.
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http://dx.doi.org/10.1016/j.vaccine.2015.02.066DOI Listing
April 2015

Development and efficacy of a novel live-attenuated QX-like nephropathogenic infectious bronchitis virus vaccine in China.

Vaccine 2015 Feb 27;33(9):1113-20. Epub 2015 Jan 27.

College of Animal Science, South China Agricultural University, Guangzhou 510642, PR China; Key Laboratory of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, Guangzhou 510642, PR China; Key Laboratory of Animal Health Aquaculture and Environmental Control, Guangzhou 510642, PR China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510640, PR China. Electronic address:

In this study, we attenuated a Chinese QX-like nephropathogenic infectious bronchitis virus (IBV) strain, YX10, by passaging through fertilized chicken eggs. The 90th passage strain (YX10p90) was selected as the live-attenuated vaccine candidate strain. YX10p90 was found to be safe in 7-day-old specific pathogen free chickens without induction of morbidity or mortality. YX10p90 provided nearly complete protection against QX-like (CH I genotype) strains and partial protection against other two major Chinese genotype strains. YX10p90 also showed no reversion to virulence after five back passages in chickens. An IBV polyvalent vaccine containing YX10p90 was developed and showed that it could provide better protection against major Chinese IBV virulent strains than commercial polyvalent vaccines. In addition, the complete genome sequence of YX10p90 was sequenced. Multiple-sequence alignments identified 38 nucleotide substitutions in the whole genome which resulted in 26 amino acid substitutions and a 110-bp deletion in the 3' untranslated region. In conclusion, the attenuated YX10p90 strain exhibited a fine balance between attenuation and immunogenicity, and should be considered as a candidate vaccine to prevent infection of Chinese QX-like nephropathogenic IBV.
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http://dx.doi.org/10.1016/j.vaccine.2015.01.036DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7127481PMC
February 2015

Identification of a chicken anemia virus variant-related gyrovirus in stray cats in china, 2012.

Biomed Res Int 2014 13;2014:313252. Epub 2014 Feb 13.

College of Animal Science, South China Agricultural University, Wushan Road, Tianhe District, Guangzhou, Guangdong 510642, China ; Key Laboratory of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, Guangzhou 510642, China.

The chicken anemia virus (CAV), is a known member of the genus Gyrovirus and was first isolated from chickens in Japan in 1979. Some reports have also demonstrated that CAV can be identified in human stool specimens. In this study, a variant of CAV was detected using PCR with CAV-based primers in fecal samples of stray cats. The genome of CAV variant was sequenced and the results suggest that it could be a recombinant viral strain from parental CAV strains JQ690762 and AF311900. Recombination is an important evolutionary mechanism that contributes to genetic diversification. These findings indicate that CAV variant might have originated from CAV-infected chickens. The epidemiology and pathogenesis of this novel virus remains to be elucidated. This study underscores the importance of CAV surveillance and it presents the first evidence suggesting the possibility of CAV homologous recombination in cat.
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http://dx.doi.org/10.1155/2014/313252DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3943257PMC
May 2015

Development of a minor groove binder assay for real-time PCR detection of porcine Sapelovirus.

J Virol Methods 2014 Mar 21;198:69-74. Epub 2013 Dec 21.

State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510006, China. Electronic address:

A 5' conjugated minor groove binder (MGB) probe real-time PCR assay was developed in this study for porcine sapelovirus (PSV) detection and quantitation. Two primers and a MGB probe for the 5' untranslated region (UTR) gene were designed. The assay was capable of detecting about 103copies/μl of standard template per reaction. Moreover, it does not detect any of the other RNA viruses that cause diarrhea disease in pigs. The coefficients of variation of intra- and inter-assay reproducibility were both lower than 2%. In 73 field fecal samples, PSV was detected in 46 samples using real-time PCR assay and only 32 samples with a conventional PCR assay. Therefore, the availability of this assay will facilitate further studies on the epidemiology of PSV infection and its role in swine disease.
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http://dx.doi.org/10.1016/j.jviromet.2013.12.003DOI Listing
March 2014

Phylogenetic diversity and genotypic complexity of H1N1 subtype swine influenza viruses isolated in mainland China.

Virol J 2012 Nov 26;9:289. Epub 2012 Nov 26.

College of Animal Science, South China Agricultural University, Guangzhou, 510642, China.

Background: After the occurrence of 2009 pandemic H1N1, close attention has been paid to the H1N1 subtype swine influenza viruses (H1N1 SIV) by scientific communities in many countries. A large-scale sequence analysis of the NCBI Influenza Virus Resource Database on H1N1 SIVs submitted primarily by scientists in China during 1992 to 2011 was performed. The aims of this study were to elucidate the genetic and evolutionary characteristics of H1N1 SIVs, to identify and unify the lineages and genetic characteristics of the H1N1 SIVs isolated in mainland China.

Results: Most of the strains were isolated during the period of 2008 to 2010 from Guangdong and Shandong provinces, China. Based on the phylogenetic and genotypic analyses, all of the H1N1 SIV strains can be classified into 8 lineages and 10 genotypes. All strains were of the characteristics of low pathogenic influenza viruses. The viruses of different lineage are characterized with different amino acid residues at the receptor-binding sites. Viruses containing PB2 genes of the classical swine, early seasonal human and recent seasonal human lineage might be more infectious to human. Some genotypes were directly related with human influenza viruses, which include strains that harbored genes derived from human influenza viruses.

Conclusions: Phylogenetic diversity and complexity existed in H1N1 SIVs isolated in mainland China. These H1N1 SIV strains were closely related to other subtype influenza viruses, especially to human influenza viruses. Moreover, it was shown that, novel lineages and genotypes of H1N1 SIVs emerged recently in mainland China. These findings provided new and essential information for further understanding of the genetic and evolutionary characteristics and monitoring the H1N1 SIVs in mainland China.
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http://dx.doi.org/10.1186/1743-422X-9-289DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3585526PMC
November 2012

Complete genome sequence of a recombinant nephropathogenic infectious bronchitis virus strain in China.

J Virol 2012 Dec;86(24):13812-3

College of Animal Science, South China Agricultural University, Guangzhou, People's Republic of China.

Recently, nephropathogenic infectious bronchitis virus (IBV) outbreaks have occurred in commercial broiler flocks and have been associated with a high incidence and morbidity in China. The CK/CH/Zhejiang/06/10 strain (IBV-YX10) was isolated from a 12-day-old broiler chicken in a flock of chickens with swollen speckled kidneys and distended ureters filled with uric acid in China in 2010. Here we reported the complete genomic sequence of the IBV-YX10 which was a natural recombinant nephropathogenic infectious bronchitis virus strain. These findings will contribute additional insights into the molecular characteristics of evolving IBV genomes and the need for effective control of IBV in China.
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http://dx.doi.org/10.1128/JVI.02575-12DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3503108PMC
December 2012

Complete genome sequence of a Newcastle disease virus strain isolated from broiler breeder flocks in China.

J Virol 2012 Nov;86(22):12461-2

College of Animal Science, South China Agricultural University, Tianhe District, Wushan Road, Guangzhou, Guangdong, People's Republic of China.

In 2010 and 2011, several devastating Newcastle disease (ND) outbreaks occurred in China, affecting broilers, layers, and breeders. The CK-JSX1-201005 virus was isolated from broiler breeder flocks vaccinated with the classical ND virus (NDV) vaccine program, but laying rate decreased from 80% to 30 to 40% in the clinic. Here, we report the complete genome sequence and molecular characteristic of the CK-JSX1-201005 NDV. These findings provide additional insights into the genetic variation of NDV circulating in China and are useful for vaccine development for NDV.
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http://dx.doi.org/10.1128/JVI.02314-12DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3486497PMC
November 2012

Complete genome sequence of porcine circovirus 2d strain GDYX.

J Virol 2012 Nov;86(22):12457-8

College of Animal Science, South China Agricultural University, Guangzhou, China.

Porcine circovirus type 2 (PCV2) is the etiologic agent of porcine circovirus-associated disease, and it is mainly divided into five genotypes. Here, we report the complete genome sequence of PCV2 strain GDYX, which belongs to PCV2d and has a unique amino acid variation at position 169 (S to G).
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http://dx.doi.org/10.1128/JVI.02290-12DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3486447PMC
November 2012

Complete genome sequence of a porcine orthoreovirus from southern China.

J Virol 2012 Nov;86(22):12456

State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, People's Republic of China.

Porcine orthoreoviruses belong to the family Reoviridae and cause mainly mild enteritis in piglets. We present here the complete genome sequence of a novel porcine orthoreovirus strain (GD-1) isolated from a piglet in southern China. Our data will facilitate future investigations of the molecular characteristics and epidemiology of porcine orthoreoviruses.
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http://dx.doi.org/10.1128/JVI.02254-12DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3486476PMC
November 2012
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