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5. Dual-Emission Fluorescence Resonance Energy Transfer (FRET) PCR Discriminates Salmonella Pullorum and Gallinarum.

Author

Gong, Jiansen, Iduu, Nneka Vivian, Zhang, Di, Chenoweth, Kelly, Wei, Lanjing, Yang, Yi, Dou, Xinhong, and Wang, Chengming

Subjects
FLUORESCENCE resonance energy transfer, SALMONELLA enterica, CONTAMINATION of poultry, SINGLE nucleotide polymorphisms, SALMONELLA diseases
Abstract

Salmonella Pullorum (S. Pullorum) and Salmonella Gallinarum (S. Gallinarum) are two biovars of Salmonella enterica serovar Gallinarum, responsible for pullorum disease and fowl typhoid, which are the most prevalent and pathogenic forms of salmonellosis in poultry in developing countries. Traditional differentiation methods for S. Pullorum and S. Gallinarum are based on distinct clinical manifestations and biochemical traits, given their indistinguishable nature via serological assays alone. Molecular differentiation methods such as allele-specific PCR and dual PCR combined with gel electrophoresis or enzyme digestion have also been used to discriminate S. Pullorum and S. Gallinarum, but the detection efficiency is not high. This investigation introduces a Fluorescence Resonance Energy Transfer (FRET) PCR assay targeting the pegB gene, exclusively found in specific Salmonella serovars such as S. Pullorum and S. Gallinarum, and exhibiting conserved single-nucleotide polymorphisms across these two biovars. High-resolution melting curve analysis demonstrates distinct dissolution profiles, facilitating the precise discrimination of S. Pullorum and S. Gallinarum. This FRET-PCR assay exhibits a detection limit of 10 copies per reaction and has been rigorously validated utilizing 17 reference strains and 39 clinical isolates. The innovation presented herein provides a valuable tool for the rapid differentiation of S. Pullorum and S. Gallinarum, thereby enhancing diagnostic efficiency and molecular surveillance of poultry Salmonella. The developed pegB-targeting FRET-PCR assay presents a promising alternative to current cumbersome and time-consuming diagnostic modalities, offering significant potential for expedited identification and control of Salmonella in poultry and mitigating economic losses associated with Salmonella contamination in poultry production. [ABSTRACT FROM AUTHOR]

Published
2024
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6. A Retrospective Analysis of Salmonella Isolates across 11 Animal Species (1982–1999) Led to the First Identification of Chromosomally Encoded blaSCO-1 in the USA

Author

Iduu, Nneka Vivian, primary, Raiford, Donna, additional, Conley, Austin, additional, Scaria, Joy, additional, Nelson, Julie, additional, Ruesch, Laura, additional, Price, Stuart, additional, Yue, Min, additional, Gong, Jiansen, additional, Wei, Lanjing, additional, and Wang, Chengming, additional

Published
2024
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12. Genome degradation promotes Salmonella pathoadaptation by remodeling fimbriae-mediated proinflammatory response

Author

Zhou, Xiao, primary, Kang, Xiamei, additional, Chen, Jiaqi, additional, Song, Yan, additional, Jia, Chenghao, additional, Teng, Lin, additional, Tang, Yanting, additional, Jiang, Zhijie, additional, Peng, Xianqi, additional, Tao, Xiaoxi, additional, Xu, Yiwei, additional, Huang, Linlin, additional, Xu, Xuebin, additional, Xu, Yaohui, additional, Zhang, Tengfei, additional, Yu, Shenye, additional, Gong, Jiansen, additional, Wang, Shaohui, additional, Liu, Yuqing, additional, Zhu, Guoqiang, additional, Kehrenberg, Corinna, additional, Weill, François-Xavier, additional, Barrow, Paul, additional, Li, Yan, additional, Zhao, Guoping, additional, and Yue, Min, additional

Published
2023
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14. A Retrospective Analysis of Salmonella Isolates across 11 Animal Species (1982–1999) Led to the First Identification of Chromosomally Encoded bla SCO-1 in the USA.

Author

Iduu, Nneka Vivian, Raiford, Donna, Conley, Austin, Scaria, Joy, Nelson, Julie, Ruesch, Laura, Price, Stuart, Yue, Min, Gong, Jiansen, Wei, Lanjing, and Wang, Chengming

Subjects
ANIMAL species, PLASMIDS, SALMONELLA enterica, SALMONELLA, WHOLE genome sequencing, MICROBIAL sensitivity tests, RETROSPECTIVE studies, DRUG resistance in microorganisms
Abstract

Antimicrobial resistance (AMR) in non-typhoidal Salmonella is a pressing public health concern in the United States, necessitating continuous surveillance. We conducted a retrospective analysis of 251 Salmonella isolates from 11 animal species recovered between 1982 and 1999, utilizing serotyping, antimicrobial susceptibility testing, and whole-genome sequencing (WGS). Phenotypic resistance was observed in 101 isolates, with S. Typhimurium, S. Dublin, S. Agona, and S. Muenster prevailing among 36 identified serovars. Notably, resistance to 12 of 17 antibiotics was detected, with ampicillin being most prevalent (79/251). We identified 38 resistance genes, primarily mediating aminoglycoside (n = 13) and β-lactamase (n = 6) resistance. Plasmid analysis unveiled nine distinct plasmids associated with AMR genes in these isolates. Chromosomally encoded blaSCO-1 was present in three S. Typhimurium and two S. Muenster isolates from equine samples, conferring resistance to amoxicillin/clavulanic acid. Phylogenetic analysis revealed three distinct clusters for these five isolates, indicating evolutionary divergence. This study represents the first report of blaSCO-1 in the USA, and our recovered isolates harboring this gene as early as 1989 precede those of all other reports. The enigmatic nature of blaSCO-1 prompts further research into its function. Our findings highlight the urgency of addressing antimicrobial resistance in Salmonella for effective public health interventions. [ABSTRACT FROM AUTHOR]

Published
2024
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19. Comprehensive Assessment of Subtyping Methods for Improved Surveillance of Foodborne Salmonella

Author

Pan, Hang, primary, Jia, Chenghao, additional, Paudyal, Narayan, additional, Li, Fang, additional, Mao, Junyong, additional, Liu, Xi, additional, Dong, Chenghang, additional, Zhou, Kun, additional, Liao, Xiayi, additional, Gong, Jiansen, additional, Fang, Weihuan, additional, Li, Xiaoliang, additional, Kehrenberg, Corinna, additional, and Yue, Min, additional

Published
2022
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21. Controlled expression of lysis gene E by a mutant of the promoter pL of the thermo‐inducible λcI857‐pL system

Author

Gong Jiansen, Bo Gao, Dejun Ji, Lixia Fu, Lingbin Zeng, and Xian’gan Han

Subjects
Gene Expression Regulation, Viral, Lysis, Mutant, Mutagenesis (molecular biology technique), medicine.disease_cause, Applied Microbiology and Biotechnology, Viral Proteins, 03 medical and health sciences, Bacteriolysis, Plasmid, Gene expression, Escherichia coli, medicine, Promoter Regions, Genetic, Gene, 030304 developmental biology, 0303 health sciences, 030306 microbiology, Chemistry, Temperature, General Medicine, Bacteriophage lambda, Molecular biology, Salmonella enteritidis, Lytic cycle, Mutation, Plasmids, Biotechnology
Abstract

AIMS: To identify a lambda promoter pL mutant that could extend the thermal stability of the thermo‐inducible λcI857‐pR/pL system and to evaluate the effects of the modified system for the controlled expression of lysis gene E during the production of bacterial ghosts (BGs). METHODS AND RESULTS: The promoter pL mutant was identified by random mutagenesis and site‐directed mutagenesis. The results showed that a T → 35C mutation in the pL promoter was responsible for the phenotype alteration. Under the same induction conditions, the lysis rates of the modified lytic system on Escherichia coli and Salmonella enteritidis were significantly lower than that of the control, while the lysis rates of Escherichia coli with the thermo‐inducible lytic system were significantly higher than that of S. enteritidis with the corresponding plasmid (P

Published
2020
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28. Efficient fecal-oral and possible vertical, but not respiratory, transmission of emerging Chlamydia gallinacea in broilers

Author

You, Jinfeng, primary, Wu, Yantao, additional, Zhang, Xiaorong, additional, Wang, Xiaobo, additional, Gong, Jiansen, additional, Zhao, Zhenhua, additional, Zhang, Jianjun, additional, Zhang, Jilei, additional, Sun, Ziqiang, additional, Li, Jing, additional, Guo, Weina, additional, Wang, Jiawei, additional, Wang, Yaoyao, additional, Li, Min, additional, Zhang, Rong, additional, Yang, Yi, additional, Ye, Jianqiang, additional, Poudel, Anil, additional, Kaltenboeck, Bernhard, additional, and Wang, Chengming, additional

Published
2019
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29. Characterization of avian pathogenic Escherichia coli isolated in eastern China

Author

Xiangan Han, Xu Ming, Zhang Di, Shen Haiyu, Zhuang Linlin, Zou Jianmin, Dou Xinhong, Gong Jiansen, and Liu Jiasheng

Subjects
0301 basic medicine, Serotype, China, Nalidixic acid, 040301 veterinary sciences, Tetracycline, 030106 microbiology, Microbial Sensitivity Tests, Drug resistance, medicine.disease_cause, Integrons, Microbiology, 0403 veterinary science, 03 medical and health sciences, Pathogenic Escherichia coli, Drug Resistance, Multiple, Bacterial, Ampicillin, Escherichia coli, Genetics, medicine, Animals, Serotyping, Escherichia coli Infections, Poultry Diseases, biology, Escherichia coli Proteins, 04 agricultural and veterinary sciences, General Medicine, bacterial infections and mycoses, biology.organism_classification, Virology, Multiple drug resistance, Biofilms, Fimbriae Proteins, Bacterial Outer Membrane Proteins, medicine.drug
Abstract

In order to investigate the biological characteristics of avian pathogenic Escherichia coli (APEC) isolated in eastern China, a total of 243 isolates were isolated from diseased poultry on different farms during the period from 2007 to 2014. These isolates were characterized for serogroups (polymerase chain reaction and agglutination), the presence of virulence-associated genes (fimC, iss, ompA, fyuA, stx2f, iroC, iucD, hlyE, tsh, cvaC, irp2, and papC) and class I integrons (polymerase chain reaction), drug susceptibilities (disk diffusion method) and the biofilm-forming abilities (semi-quantitative method). The results showed that the most predominant serogroups were O78 (87 isolates, 35.8%) and O2 (35 isolates, 14.4%). Gene profiling found that fimC and ompA were frequently distributed among the isolates and that 77.4% of the isolates were positive for class 1 integrons. Overall, isolates displayed resistance to tetracycline (97.5%), nalidixic acid (82.3%), ampicillin (81.1%), sulphafurazole (80.7%), streptomycin (79.0%), trimethoprim (78.2%) and cotrimoxazole (78.2%). Multiple-drug resistance was exhibited in 80.3% of the isolates, and the presence of class 1 integrons is associated with multidrug resistance. Finally, 151 isolates had the ability to form biofilms in vitro, and drug resistance seemed relative to biofilm-forming abilities.

Published
2016
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34. Newly identified colistin resistance genes, mcr-4 and mcr-5, from upper and lower alimentary tract of pigs and poultry in China

Author

Chen, Li, primary, Zhang, Jilei, additional, Wang, Jiawei, additional, Butaye, Patrick, additional, Kelly, Patrick, additional, Li, Min, additional, Yang, Feng, additional, Gong, Jiansen, additional, Yassin, Afrah Kamal, additional, Guo, Weina, additional, Li, Jing, additional, Song, Chunlian, additional, and Wang, Chengming, additional

Published
2018
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35. Molecular detection of colistin resistance genes (mcr-1, mcr-2 and mcr-3) in nasal/oropharyngeal and anal/cloacal swabs from pigs and poultry

Author

Zhang, Jilei, primary, Chen, Li, additional, Wang, Jiawei, additional, Yassin, Afrah Kamal, additional, Butaye, Patrick, additional, Kelly, Patrick, additional, Gong, Jiansen, additional, Guo, Weina, additional, Li, Jing, additional, Li, Min, additional, Yang, Feng, additional, Feng, Zhixing, additional, Jiang, Ping, additional, Song, Chunlian, additional, Wang, Yaoyao, additional, You, Jinfeng, additional, Yang, Yi, additional, Price, Stuart, additional, Qi, Kezong, additional, Kang, Yuan, additional, and Wang, Chengming, additional

Published
2018
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38. From genomes to genotypes: molecular epidemiological analysis of Chlamydia gallinacea reveals a high level of genetic diversity for this newly emerging chlamydial pathogen

Author

Guo, Weina, primary, Jelocnik, Martina, additional, Li, Jing, additional, Sachse, Konrad, additional, Polkinghorne, Adam, additional, Pannekoek, Yvonne, additional, Kaltenboeck, Bernhard, additional, Gong, Jiansen, additional, You, Jinfeng, additional, and Wang, Chengming, additional

Published
2017
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39. Identification and characterization of mcr mediated colistin resistance in extraintestinal Escherichia coli from poultry and livestock in China

Author

Yassin, Afrah Kamal, primary, Zhang, Jilei, additional, Wang, Jiawei, additional, Chen, Li, additional, Kelly, Patrick, additional, Butaye, Patrick, additional, Lu, Guangwu, additional, Gong, Jiansen, additional, Li, Min, additional, Wei, Lanjing, additional, Wang, Yaoyao, additional, Qi, Kezong, additional, Han, Xiangan, additional, Price, Stuart, additional, Hathcock, Terri, additional, and Wang, Chengming, additional

Published
2017
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43. Highly Drug-Resistant Salmonella enterica Serovar Indiana Clinical Isolates Recovered from Broilers and Poultry Workers with Diarrhea in China

Author

Gong, Jiansen, primary, Wang, Chengming, additional, Shi, Shourong, additional, Bao, Hongduo, additional, Zhu, Chunhong, additional, Kelly, Patrick, additional, Zhuang, Linlin, additional, Lu, Guangwu, additional, Dou, Xinhong, additional, Wang, Ran, additional, Xu, Bu, additional, and Zou, Jianmin, additional

Published
2016
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44. Cloning of the genome of a goose parvovirus vaccine strain SYG61v and rescue of infectious virions from recombinant plasmid in embryonated goose eggs

Author

Xia Meng, Gong Jiansen, Jinkun Duan, Guoqiang Zhu, Jianye Wang, and Zhiwei Jiang

Subjects
China, Zygote, Molecular Sequence Data, Virulence, Vaccines, Attenuated, Genome, Virus, law.invention, Parvovirus, Goose, Plasmid, law, Virology, biology.animal, Geese, Animals, Cloning, Molecular, pBluescript, Polymorphism, Genetic, biology, Embryonated, Virion, Viral Vaccines, Sequence Analysis, DNA, DNA, Viral, Mutation, Recombinant DNA, Sequence Alignment
Abstract

The SYG61v is an attenuated goose parvovirus (GPV) that has been used as a vaccine strain in China. The genome of SYG61v was sequenced to attempt to identify the genetic basis for the attenuation of this strain. The entire genome consists of 5102 nucleotides (nts), with four nt deletions compared to that of virulent strain B. The inverted terminal repeats (ITR) are 442 nts in length, of which 360 nts form a stem region, and 43 nts constitute the bubble region. Although mutations were observed throughout the ITR, no mismatch was found in the stem. Alignment with other pathogenic GPV strains (B, 82-0321, 06-0329, and YZ99-5) indicated that there are 10 and 11 amino acid mutations in the Rep1 and VP1 proteins of SYG61v, respectively. The complete genome of SYG61v was cloned into the pBluescript II vector and an infectious plasmid pSYG61v was generated. Infectious progeny virus was successfully rescued through transfection of the plasmid pSYG61v in embryonated goose eggs and yielded viral titers similar to its parental virus, as evaluated by ELD50.

Published
2013

45. SEF14 fimbriae from Salmonella enteritidis play a role in pathogenitic to cell model in vitro and host in vivo

Author

Huayan Hou, Gong Jiansen, Guoqiang Zhu, Xia Meng, Zhiyun Tao, Chunhong Zhu, and Xiaoli Duan

Subjects
Mice, Inbred BALB C, Salmonella Infections, Animal, Virulence, Swine, Virulence Factors, Salmonella enteritidis, Fimbria, Mutant, Epithelial Cells, Biology, Microbiology, Median lethal dose, In vitro, Lethal Dose 50, Mice, Infectious Diseases, In vivo, Cell culture, Macrophages, Peritoneal, Animals, Fimbriae Proteins, Cells, Cultured
Abstract

The role of SEF14 fimbriae in virulence remains to be elucidated and in this study, we showed that sefA mutant constructed in the wild-type (WT) Salmonella enteritidis strain 50336 displayed increased invasion to IPEC-J2 cell lines and survival in mouse peritoneal macrophages, and the lethal dose 50% (LD50) in 6-week-old Balb/c mice intra-peritoneally injected with WT S. enteritidis strain decreased significantly upon deletion of sefA indicating their role in virulence. Overall, these results demonstrated that expression of sefA of SEF14 fimbriae enhances S. enteritidis adhesion to epithelial cells and survival in macrophages and contributes to S. enteritidis virulence in mice.

Published
2013

49. Serovar-Specific Polymerase Chain Reaction for Detection of Salmonella entericaSerovar Indiana

Author

Zhang, Ping, Zhuang, Linlin, Zhang, Di, Xu, Jingxiao, Dou, Xinhong, Wang, Chengming, and Gong, Jiansen

Abstract

AbstractSalmonella entericaserovar Indiana (S.Indiana) is a newly emerging pathogen with high levels of drug resistance. It has become one of the most common Salmonellaserovars in China with a worldwide distribution, posing significant public health concerns. Detection of S. Indiana by traditional bacteriological methods is time-consuming and laborious, which prevents timely surveillance and effective control of the pathogen. In this study, comparative genomics was used to identify an A7P63_13850 gene that is uniquely present in S. Indiana, but not in other Salmonellaserovars or any non-Salmonellabacteria. Then, a polymerase chain reaction (PCR) assay targeting this serovar-specific gene was established for specific detection of S. Indiana. The detection limit of this method is 10 pg per reaction for bacterial genomic DNA, being equivalent to 100 colony-forming units (CFU) per reaction. The established PCR amplifies all S. Indiana strains (n= 56), but none of other Salmonellaserovars (n= 146) and non-Salmonellaspecies (n= 14). The assay established in this study was also used to detect clinical samples from poultry, showed a positivity of 14.7% (23/156) for S. Indiana, which were verified by bacteriological methods. The highly sensitive and serovar-specific PCR for S. Indiana established in this study is suitable and convenient for detection of S. Indiana which aids in surveillance and control of the pathogen.

Published
2018
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50. Establishment of a Multiplex Loop-Mediated Isothermal Amplification Method for Rapid Detection of Sulfonamide Resistance Genes (sul1, sul2, sul3) in Clinical EnterobacteriaceaeIsolates from Poultry

Author

Gong, Jiansen, Zhuang, Linlin, Zhang, Di, Zhang, Ping, Dou, Xinhong, and Wang, Chengming

Abstract

AbstractAntimicrobial resistance genes play an important role in mediating resistance to sulfonamide in Gram-negative bacteria. While PCR is the current method to detect sulfonamide resistance genes (sul1, sul2, sul3), it is time-consuming and costly and there is an urgent need to develop a more convenient, simpler and rapid test for the sul. In this study, we describe a multiplex loop-mediated isothermal amplification (m-LAMP) assay we developed for the rapid and simultaneous detection of three sul. This m-LAMP assay successfully detected seven reference strains with different sulgenotypes, but was negative for nine sul-negative reference strains. The m-LAMP products were verified by HinfI restriction enzyme digestion and the detection limit of the test was 0.5 pg genomic DNA per reaction. Testing 307 sulfonamide-resistant Enterobacteriaceaeclinical isolates with the m-LAMP revealed all were positive for the sulwith sul2(79.5%) and sul1(64.5%) being most prevalent, and sul3the least (12.1%). Of the Enterobacteriaceaeisolates tested, the SalmonellaIndiana, a newly emerging serovar resistant to numerous antimicrobials, were most commonly positive with 33% having sul3.

Published
2018
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