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Generation of double knockout cattle via CRISPR-Cas9 ribonucleoprotein(RNP)electroporation

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Journal of Animal Science and Biotechnology 2024年第1期15卷 456-462页

作者： Gyeong-Min Gim Kyeong-Hyeon Eom Dong-Hyeok Kwon Dae-Jin Jung Dae-Hyun Kim Jun-Koo Yi Jae-Jung Ha Ji-Hyun Lee Seong-Beom Lee Woo-Jae Son Soo-Young Yum Won-Wu Lee Goo Jang Laboratory of Theriogenology and Biotechnology Department of Veterinary Clinical ScienceCollege of Veterinary Medicine and the Research Institute of Veterinary ScienceSeoul National UniversitySeoulRepublic of Korea LARTBio Co. Ltd.SeoulRepublic of Korea Gyeongsangbukdo Livestock research Institute YeongjuRepublic of Korea Comparative Medicine disease research center Seoul National UniversitySeoulRepublic of Korea

Background Genome editing has been considered as powerful tool in agricultural ***,genome editing progress in cattle has not been fast as in other mammal species,for some disadvantages including long gestational periods,single pregnancy,and high raising ***,technically demanding methods such as microinjection and somatic cell nuclear transfer(SCNT)are needed for gene editing in *** this point of view,electroporation in embryos has been risen as an *** First,editing efficiency of our electroporation methods were tested for *** of mutation on embryo was confirmed by T7E1 *** first combination,mutation rates for MSTN and PRNP were 57.6%±13.7%and 54.6%±13.5%,*** case of MSTN/BLG,mutation rates were 83.9%±23.6%for MSTN,84.5%±18.0%for ***,the double-KO embryos were transferred to surrogates and mutation rate was identified in resultant calves by targeted deep *** recipients were transferred for MSTN/PRNP,4 calves were delivered,and one calf underwent an induction for double *** surrogates were given double-KO embryos for MSTN/BLG,and four of the six calves that were born had mutations in both *** These data demonstrated that production of genome edited cattle via electroporation of RNP could be effectively ***,MSTN and PRNP from beef cattle and MSTN and BLG from dairy cattle have been born and they will be valuable resources for future precision breeding.

关键词： Beta-lactoglobulin Cattle CRISPR-Cas9 Electroporation Knockout MSTN PRNP

Genetic and pathogenic characterization of new infectious bronchitis virus strains in the GVI-1 and GI-19 lineages isolated in central China

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Journal of Integrative Agriculture 2024年第7期23卷 2407-2420页

作者： Yuhan Yang Dou Wang Yaning Bai Wenyan Huang Shimin Gao Xingchen Wu Ying Wang Jianle Ren Jinxin He Lin Jin Mingming Hu Zhiwei Wang Zhongbing Wang Haili Ma Junping Li Libin Liang College of veterinary Medicine Shanxi Agricultural UniversityJinzhong 030801China Shanxi Provincial Animal disease Prevention and Control center Taiyuan 030000China

Avian infectious bronchitis(IB)is a highly contagious infectious disease caused by infectious bronchitis virus(IBV),which is prevalent in many countries worldwide and causes serious harm to the poultry *** present,many commercial IBV vaccines have been used for the prevention and control of IB;however,IB outbreaks occur *** this study,two new strains of IBV,SX/2106 and SX/2204,were isolated from two flocks which were immunized with IBV H120 vaccine in central *** and recombination analysis indicated that SX/2106,which was clustered into the GI-19 lineage,may be derived from recombination events of the GI-19 and GI-7 strains and the LDT3-A *** analysis showed that SX/2204 belongs to the GVI-1 lineage,which may have originated from the recombination of the GI-13 and GVI-1 strains and the H120 *** virus cross-neutralization test showed that the antigenicity of SX/2106 and SX/2204 was different from *** experiments found that both SX/2106 and SX/2204 could replicate effectively in the lungs and kidneys of chickens and cause disease and death,and H120 immunization could not provide effective protection against the two IBV *** is noteworthy that the pathogenicity of SX/2204 has significantly increased compared to the GVI-1 strains isolated previously,with a mortality rate up to 60%.Considering the continuous mutation and recombination of the IBV genome to produce new variant strains,it is important to continuously monitor epidemic strains and develop new vaccines for the prevention and control of IBV epidemics.

关键词： infectious bronchitis virus GI-19 lineage GVI-1 lineage complete genome recombination pathogenicity

Genetic and biological properties of H9N2 avian influenza viruses isolated in central China from2020 to 2022

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Journal of Integrative Agriculture 2024年第8期23卷 2778-2791页

作者： Libin Liang Yaning Bai Wenyan Huang Pengfei Ren Xing Li Dou Wang Yuhan Yang Zhen Gao Jiao Tang Xingchen Wu Shimin Gao Yanna Guo Mingming Hu Zhiwei Wang Zhongbing Wang Haili Ma Junping Li College of veterinary Medicine Shanxi Agricultural UniversityJinzhong 030801China Shanxi Provincial Animal disease Prevention and Control center Taiyuan 030000China

The H9N2 subtype of avian influenza virus(AIV)is widely prevalent in poultry and wild birds globally,and has become the predominant subtype circulating in poultry in *** H9N2 AIV can directly or indirectly(by serving as a"donor virus")infect humans,posing a significant threat to public ***,there is a lack of in-depth research on the prevalence of H9N2 viruses in Shanxi Province,central *** this study,we isolated 14 H9N2 AIVs from October 2020 to April 2022 in Shanxi Province,and genetic analysis revealed that these viruses belonged to 7 different *** study on animals revealed that the H9N2 strains we identified displayed high transmission efficiency among chicken populations,and exhibited diverse replication abilities within these *** viruses could replicate efficiently in the lungs of mice,with one strain also demonstrating the capacity to reproduce in organs like the brain and *** the cellular level,the replication ability of different H9N2 strains was evaluated using plaque formation assays and multi-step growth curve assays,revealing significant differences in the replication and proliferation efficiency of the various H9N2 viruses at the cellular *** antigenicity analysis suggested that these isolates could be classified into 2 separate antigenic *** research provides crucial data to help understand the prevalence and biological characteristics of H9N2 AIVs in central *** also highlights the necessity of enhancing the surveillance of H9N2 AIVs.

关键词： avian influenza virus H9N2 central China pathogenicity antigenicity

A novel live attenuated vaccine candidate protects chickens against subtype B avian metapneumovirus

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Journal of Integrative Agriculture 2024年第5期23卷 1658-1670页

作者： Lingzhai Meng Mengmeng Yu Suyan Wang Yuntong Chen Yuanling Bao Peng Liu Xiaoyan Feng Tana He Ru Guo Tao Zhang Mingxue Hu Changjun Liu Xiaole Qi Kai Li Li Gao Yanping Zhang Hongyu Cui Yulong Gao Avian Immunosuppressive diseases Division State Key Laboratory for Animal Disease Control and PreventionHarbin Veterinary Research InstituteChineseAcademy of Agricultural SciencesHarbin150069China Avian ImmunosuppressiveDiseases DivisionStateKey LaboratoryforAnimal Disease Cont National Poultry Laboratory Animal Resource center Harbin 150069China

Avian metapneumovirus(aMPV) is a highly contagious pathogen that causes acute upper respiratory tract diseases in chickens and turkeys, resulting in serious economic losses. Subtype B aMPV has recently become the dominant epidemic strain in China. We developed an attenuated aMPV subtype B strain by serial passaging in Vero cells and evaluated its safety and efficacy as a vaccine candidate. The safety test showed that after the 30th passage, the LN16-A strain was fully attenuated, as clinical signs of infection and histological lesions were absent after *** LN16-A strain did not revert to a virulent strain after five serial passages in chickens. The genomic sequence of LN16-A differed from that of the parent wild-type LN16(wtLN16) strain and had nine amino acid mutations. In chickens, a single immunization with LN16-A induced robust humoral and cellular immune responses, including the abundant production of neutralizing antibodies, CD4^(+) T lymphocytes, and the Th1(IFN-γ) and Th2(IL-4 and IL-6)cytokines. We also confirmed that LN16-A provided 100% protection against subtype B aMPV and significantly reduced viral shedding and turbinate inflammation. Our findings suggest that the LN16-A strain is a promising live attenuated vaccine candidate that can prevent infection with subtype B aMPV.

关键词： avian metapneumovirus attenuated protection vaccine candidate chickens

Emergence of a novel multi-resistance-mediating integrative and conjugative element ICEPmu3 in Pasteurella multocida

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Journal of Integrative Agriculture 2024年第11期23卷 3938-3942页

作者： Jiao He Zhishuang Yang Mingshu Wang Renyong Jia Shun Chen Mafeng Liu Xinxin Zhao Qiao Yang Ying Wu Shaqiu Zhang Juan Huang Xumin Ou Di Sun Bin Tian Yu He Zhen Wu Anchun Cheng Dekang Zhu research center of Avian diseases College of Veterinary MedicineSichuan Agricultural University International Joint research center for Animal disease Prevention and Control of Sichuan Province Engineering research center of Southwest Animal disease Prevention and Control Technology Ministry of Education Key Laboratory of Agricultural Bioinformatics Ministry of Education

Pasteurella multocida (PM) has been infecting a variety of hosts for a long time,causing sustained economic losses worldwide;however,there have been limited studies on its extensive adaptability (Aktories et al.2012).Analysis of strains data collected in our laboratory revealed that PM typically acquires foreign genes through transformation and conjugation,rather than transformation and *** and conjugative elements (ICEs)are a crucial mechanism that leads to abrupt changes in niche preferences and enhances environmental adaptability for bacteria,with their number far exceeding the number of plasmids and phages (Wozniak and Waldor 2010;Johnson and Grossman 2015;Botelho and Schulenburg 2021).Previous research has shown that the European bovine-sourced strain Pm36950 contains the experimentally transferable resistant ICEPmu1,which could integrate into the host chromosome or forming a circular intermediate (Michael et al.2012).Additionally,a few studies have suggested that PM strains isolated from European bovine and Asian swine might contain ICEs,but these claims lack experimental verification (Klima et al.2014;Moustafa et al.2015;Kadlec et al.2017;Peng et al.2017;Beker et al.2018;Schink et al.2022).Currently,there is no data available on ICE-carrying PM strains isolated from hosts outside of European cattle or Chinses *** letter presents a report on a novel ICE identified in the hypervirulent and multidrug-resistant PM HN141014 strain isolated from Chinese *** ICE was specifically analyzed for its resistance genes,transferable capacity and host diversity.

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An Outbreak of Serogroup Y Meningococcal Meningitis in a Private Secondary Vocational School-Guangzhou City,Guangdong Province,China,2023

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China CDC weekly 2024年第12期6卷 242-246,I0005页

作者： Xiang Zeng Qing He Yilan Li Yong Zhou Xiaoping Shao Pei Hu Jian Liang Yudan Song Chao Ma Lijie Zhang Limei Sun Lei Luo Chinese Field Epidemiology Training Program Chinese Center for Disease Control and PreventionBeijingChina Zhuhai center for disease Control and Prevention Zhuhai CityGuangdong ProvinceChina Guangdong center for disease Control and Prevention Guangzhou CityGuangdong ProvinceChina Guangzhou center for disease Control and Prevention Guangzhou CityGuangdong ProvinceChina Institute of Public Health Guangzhou Medical University&Guangzhou Center for Disease Control and PreventionGuangzhou CityGuangdong ProvinceChina Department of the National Immunization Programme Chinese Center for Disease Control and PreventionBeijingChina

What is already known about this topic?The inclusion of meningococcal vaccines in the National Immunization Program(NIP)over several years has significantly reduced the incidence of meningococcal meningitis in China to historic ***,there has been a diversification of meningococcal serogroups,leading to a shift in dominant serogroups in China from serogroup A to serogroups C and B,accompanied by a rise in reports of serogroups Y and *** is added by this report?An outbreak of serogroup Y Neisseria meningitidis(Nm)in a secondary vocational school involved a single confirmed severe case and 24 individuals with laboratory-confirmed Nm *** investigation revealed that the outbreak was localized to the classroom of the confirmed *** close contact within a confined space was identified as a significant risk factor for Nm *** genotype sequence identified was type 1655(ST-1655),which is categorized under clonal complex 23(CC-23)and bears resemblance to 8 previously confirmed cases of serogroup Y meningococcal meningitis within Guangdong *** suggests that serogroup Y infections continue to sporadically emerge and have become prevalent *** are the implications for public health practice?This outbreak underscores the critical need to enhance surveillance of meningococcal serogroups and population carrier,and advocate for vaccination with MenY-containing vaccines.

关键词： break Secondary Guangdong

The virulence regulator AbsR in avian pathogenic Escherichia coli has pleiotropic effects on bacterial physiology

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Journal of Integrative Agriculture 2024年第2期23卷 649-668页

作者： Dongfang Zhao Haobo Zhang Xinyang Zhang Fengwei Jiang Yijing Li Wentong Cai Ganwu Li State Key Laboratory for Animal disease Control and Prevention Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150069China College of veterinary Medicine Northeast Agricultural UniversityHarbin 150038China National Animal Tuberculosis Reference Laboratory Division of Zoonoses SurveillanceChina Animal Health and Epidemiology CenterQingdao 266032China Department of veterinary Diagnostic and Production Animal Medicine College of Veterinary MedicineIowa State UniversityAmesIowa 50011-1134the United States

Avian pathogenic Escherichia coli(APEC)belonging to extraintestinal pathogenic ***(ExPEC)can cause severe infections in extraintestinal tissues in birds and humans,such as the lungs and ***(microcin production regulation,locus A,herein renamed AbsR,a blood survival regulator),a member of the MarR(multiple antibiotic resistance regulator)transcriptional regulator family,governs the expression of capsule biosynthetic genes in human ExPEC and represents a promising druggable target for ***,a deep understanding of the AbsR regulatory mechanism as well as its regulon is *** this study,we present a systems-level analysis of the APEC AbsR regulon using ChIP-Seq(chromatin immunoprecipitation sequencing)and RNA-Seq(RNA sequencing)*** found that AbsR directly regulates 99 genes and indirectly regulates 667 ***,we showed that:1)AbsR contributes to antiphagocytotic effects by macrophages and virulence in a mouse model for systemic infection by directly activating the capsular gene cluster;2)AbsR positively impacts biofilm formation via direct regulation of the T2SS(type II secretion system)but plays a marginal role in virulence;and 3)AbsR directly upregulates the acid tolerance signaling system EvgAS to withstand acid stress but is dispensable in ExPEC ***,our data indicate that the role of AbsR in virulence gene regulation is relatively conserved in ExPEC ***,this study provides a comprehensive analysis of the AbsR regulon and regulatory mechanism,and our data suggest that AbsR likely influences virulence primarily through the control of capsule ***,we found that AbsR severely represses the expression of the type I-F CRISPR(clustered regularly interspaced short palindromic repeats)-Cas(CRISPR associated)systems,which could have implications in CRISPR biology and application.

关键词： avian pathogenic Escherichia coli(APEC) extraintestinal pathogenic Escherichia coli(ExPEC) AbsR RNA-Seq ChIP-Seq gene regulation

First complete genomic sequence analysis of canine distemper virus in wild boar

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Virologica Sinica 2024年第4期39卷 702-704页

作者： Tong Wang Haiying Du Na Feng Yuhang Liu Yu Xu Heting Sun Peng Peng Siyuan Qin Xiaotian Zhang Yan Liu Mingyuan Yu Hongrui Liang Biao He Guoqiang Zhu Changchun Tu Zhongzhong Tu Jiangsu Co-Innovation center for the Prevention and Control of Important Animal Infectious disease and Zoonoses Yangzhou UniversityYangzhou 225009China Changchun veterinary research Institute Chinese Academy of Agricultural SciencesChangchun 130122China Biological Disaster Control and Prevention center National Forestry and Grassland AdministrationShenyang 110034China

Dear Editor,Canine distemper virus(CDV)is the causal agent of a highly contagious viral infectious disease that affects domestic and wild carnivores *** is an enveloped,non-segmented negative sense RNA virus that belongs to the Morbillivirus genus in Paramyxoviridae family,which contains viruses of epidemiological relevance to humans and *** on the variability of the hemagglutinin gene(H),CDV strains have been at least 21 major genetic lineages:America-1 to America-5,Canada-1 and-2,Asia-1 to Asia-6,Europe wild-life,Arctic,Africa-1/South Africa,Africa-2,South America-1 to South America-3 and Rockborn-like(Giacinti et al.,2022).Mutations in the binding sites of the H protein,which interact with viral entry receptors such as signaling lymphocytic activation molecule(SLAM)and nectin-4,are associated with the emergence of the disease in new host species(Beineke et al.,2015).

关键词： distemper Canada globally

Neglected Time Intervals Before “1-3-7” Approach About Imported Malaria Cases-China, 2014–2021

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China CDC weekly 2024年第17期6卷 363-367页

作者： Zhiquan He Yuanjing Kou Dan Wang Ying Liu Xiaofeng Lyu Hui Yan Wen Lin Xuan Zhang Duoquan Wang Shizhu Li Hongwei Zhang Henan Provincial center for disease Control and Prevention Zhengzhou CityHenan ProvinceChina Anhui Provincial center for disease Control and Prevention Hefei CityAnhui ProvinceChina Guangxi Zhuang autonomous Region center for disease Control and Prevention Nanning CityGuangxi Zhuang Autonomous RegionChina Hubei Provincial center for disease Control and Prevention Wuhan CityHubei ProvinceChina Zhejiang Provincial center for disease Control and Prevention Hangzhou CityZhejiang ProvinceChina National Institute of Parasitic diseases Chinese Center for Disease Control and Prevention(Chinese Center for Tropical Diseases Research)National Health Commission Key Laboratory of Parasite and Vector BiologyShanghaiChina School of Global Health Chinese Center for Tropical Diseases ResearchShanghai Jiao Tong University School of MedicineShanghaiChina

What is already known about this topic?China’s“1-3-7”approach outlines specific targets to guide and monitor the processes of case reporting,investigation,and ***,few studies have examined the time intervals preceding the initial step,and the timeline from the arrival of imported malaria cases in China to their diagnosis has been largely *** is added by this report?The study demonstrated that the median duration from arrival in China to the onset of symptoms for *** was 78 days,with 71.59%of imported cases manifesting symptoms after more than one *** ***,the median interval was 42 days,with 55.91%exceeding one ***,the median time from symptom onset to malaria treatment in China between 2014 and 2021 was 2 days,with an interquartile range(IQR)of 1-4 *** are the implications for public health practice?This study represents the initial effort to delineate the chronology of imported malaria cases,from their arrival in China to their subsequent *** results underscore the importance of providing malaria health education to populations arriving from ***,enhancing physician training is crucial for improving the diagnosis of malaria.

关键词： diagnosis Malaria exceeding