1.Construction and characterization of Actinobacillus pleuropneumoniae serovar 7 live attenuated vaccine strain co-expressing ApxIA.
Jinlin LIU ; Yan CHEN ; Linlin HU ; Weicheng BEI ; Huanchun CHEN
Chinese Journal of Biotechnology 2010;26(3):305-310
Actinobacillus pleuropneumoniae (A. pleuropneumoniae), the causative agent of porcine contagious pleuropneumonia (PCP), is a significant pathogen of the world pig industry, vaccination is potentially an effective tool for the prevention of PCP. The purpose of present study was to enhance the immunogenicity of A. pleuropneumoniae live vaccine strain HB04C- (serovar 7), which was unable to express ApxIA, and to develop effective multivalent vaccines for the respiratory pathogens based on the attenuated A. pleuropneumoniae. We introduced a shuttle vector containing intact apxIA gene into HB04C-, generating HB04C2, an A. pleuropneumoniae serovar 7 live attenuated vaccine strain co-expressing ApxIA. Then we investigated the biological characteristics of HB04C2. We found that the shuttle vector expressing ApxIA was stable in HB04C2, and the growth ability of HB04C2 was not affected by the shuttle vector. We observed that HB04C2 elicited detectable antibodies against ApxIA and ApxIIA when it was administrated intratracheally as a live vaccine in pigs, and all immunized pigs were protected from heterologous virulent A. pleuropneumoniae (serovar 1) challenge. In conclusion, we demonstrated that A. pleuropneumoniae live vaccine could be used as a vector for expression of heterologous antigens.
Actinobacillus Infections
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prevention & control
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veterinary
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Actinobacillus pleuropneumoniae
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classification
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immunology
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Animals
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Bacterial Proteins
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biosynthesis
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genetics
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Bacterial Vaccines
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biosynthesis
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immunology
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Hemolysin Proteins
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biosynthesis
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genetics
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Pleuropneumonia
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microbiology
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prevention & control
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Swine
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Swine Diseases
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microbiology
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prevention & control
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Vaccines, Attenuated
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biosynthesis
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immunology
2.Assessing PCV2 antibodies in field pigs vaccinated with different porcine circovirus 2 vaccines using two commercial ELISA systems.
Min Kyoung SHIN ; Seung Hyun YOON ; Myung Hwui KIM ; Young Soo LYOO ; Seung Won SUH ; Han Sang YOO
Journal of Veterinary Science 2015;16(1):25-29
Porcine circovirus type 2 (PCV2) is the primary causative agent for post-weaning, multisystemic, wasting syndrome. Consequently, serologic detection of and vaccination against PCV2 are important for the swine industry. Among several serological tests, the enzyme-linked immunosorbent assay (ELISA) is commonly used to measure anti-PCV2 antibody levels. In the present study, we used two commercial ELISA systems to comparatively evaluate anti-PCV2 antibodies in field pigs treated with three different PCV2 vaccines. Among a total of 517 serum samples, the results of the two ELISAs were fully concordant for 365 positive and 42 negative samples, indicating 78.7% agreement. In addition, the Pearson coefficient (0.636) indicated a moderate correlation between data from the two ELISAs. Results from the farms with pigs vaccinated with the three different PCV2 vaccines demonstrated that most of the vaccinated animals underwent seroconversion. However, the increase and duration of antibody titers varied depending on the vaccine, the presence of maternal antibodies, and the vaccination program. PCV2 serologic status and anti-PCV2 antibody levels of herds from this study could be utilized to determine the best timing for vaccination and assessing vaccination compliance.
Aging
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Animals
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Antibodies, Viral/*blood
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Circovirus/*classification/immunology
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Enzyme-Linked Immunosorbent Assay/methods/*veterinary
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Female
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Porcine Postweaning Multisystemic Wasting Syndrome/blood/immunology/*prevention & control
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Republic of Korea/epidemiology
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Swine
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Swine Diseases/*prevention & control/virology
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Viral Vaccines/*immunology
3.Protective efficacy of a high-growth reassortant swine H3N2 inactivated vaccine constructed by reverse genetic manipulation.
Feng WEN ; Ji Hong MA ; Hai YU ; Fu Ru YANG ; Meng HUANG ; Yan Jun ZHOU ; Ze Jun LI ; Guang Zhi TONG
Journal of Veterinary Science 2014;15(3):381-388
Novel reassortant H3N2 swine influenza viruses (SwIV) with the matrix gene from the 2009 H1N1 pandemic virus have been isolated in many countries as well as during outbreaks in multiple states in the United States, indicating that H3N2 SwIV might be a potential threat to public health. Since southern China is the world's largest producer of pigs, efficient vaccines should be developed to prevent pigs from acquiring H3N2 subtype SwIV infections, and thus limit the possibility of SwIV infection at agricultural fairs. In this study, a high-growth reassortant virus (GD/PR8) was generated by plasmid-based reverse genetics and tested as a candidate inactivated vaccine. The protective efficacy of this vaccine was evaluated in mice by challenging them with another H3N2 SwIV isolate [A/Swine/Heilongjiang/1/05 (H3N2) (HLJ/05)]. Prime and booster inoculation with GD/PR8 vaccine yielded high-titer serum hemagglutination inhibiting antibodies and IgG antibodies. Complete protection of mice against H3N2 SwIV was observed, with significantly reduced lung lesion and viral loads in vaccine-inoculated mice relative to mock-vaccinated controls. These results suggest that the GD/PR8 vaccine may serve as a promising candidate for rapid intervention of H3N2 SwIV outbreaks in China.
Animals
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Female
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Influenza A Virus, H3N2 Subtype/*genetics/immunology
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Influenza Vaccines/genetics/immunology/*therapeutic use
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Mice
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Mice, Inbred BALB C
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Orthomyxoviridae Infections/immunology/*prevention & control/virology
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Reassortant Viruses/genetics/immunology
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Reverse Genetics/methods/*veterinary
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Swine
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Swine Diseases/immunology/*prevention & control/virology
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Vaccines, Inactivated
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Virus Replication
4.Immune Response of Recombinant Pseudorabies Virus rPRV-VP2 Expressing VP2 Gene of Porcine Parvovirus in Mice.
Pengfei FU ; Xinlong PAN ; Qiao HAN ; Xingwu YANG ; Qianlei ZHU ; Xiaoqing GUO ; Yu ZHANG ; Hongying CHEN
Chinese Journal of Virology 2016;32(2):195-202
In order to develop a combined live vaccine that will be used to prevent against porcine parvovirus (PPV) and Pseudorabies virus (PRV) infection, the VP2 gene of PPV was inserted into the transfer vector plasmid pG to produce the recombinant plasmid pGVP2. The plasmid pGVP2 and the genome of PRV HB98 attenuated vaccine were transfected by using lipofectamine into swine testis cells for the homologous recombination. The recombinant virus rPRV-VP2 was purified by selection of green fluorescence plaques for five cycles. 6-week-old female Kunming mice were immunized intramuscularly with attenuated PRV parent HB98 strain, commercial inactivated vaccine against PPV, recombinant virus, DMEM culture solution. The injections were repeated with an equivalent dose after 2 weeks in all of the groups, and then challenged with the virulent PRV NY strain at 7 weeks after the first immunization. The recombinant virus rPRV-VP2 was successfully generated, and the recombinant virus could effectively elicite anti-PPV and PRV antibody and significant cellular immune response as indicated by anti-PPV ELISA and HI, PRV-neutralizing assay and flow cytometry. The challenge assay indicated that recombinant virus could protect the mice against the virulent PRV challenge. These results demonstrated that the recombinant virus can be a candidate recombinant vaccine strain for the prevention of PRV and PPV.
Animals
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Antibodies, Viral
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immunology
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Antigens, Viral
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administration & dosage
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genetics
;
immunology
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Capsid Proteins
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administration & dosage
;
genetics
;
immunology
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Female
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Gene Expression
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Genetic Vectors
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genetics
;
metabolism
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Herpesvirus 1, Suid
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genetics
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metabolism
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Mice
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Parvovirus, Porcine
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genetics
;
immunology
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Swine
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Swine Diseases
;
immunology
;
prevention & control
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virology
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Viral Vaccines
;
administration & dosage
;
genetics
;
immunology
5.Dietary germanium biotite supplementation enhances the induction of antibody responses to foot-and-mouth disease virus vaccine in pigs.
Jin A LEE ; Bock Gie JUNG ; Myunghwan JUNG ; Tae Hoon KIM ; Han Sang YOO ; Bong Joo LEE
Journal of Veterinary Science 2014;15(3):443-447
We evaluated the potential ability of germanium biotite (GB) to stimulate the production of antibodies specific for foot-and-mouth disease virus (FMDV). To this aim, we measured the total FMDV-specific antibody responses and IgM production after vaccination against FMD both experimentally and in the field. GB supplementation with FMDV vaccination stimulated the production of anti-FMDV antibodies, and effectively increased IFN-gamma and TNF-alpha levels. These results suggest that GB may be a novel alternative feed supplement that can serve as a boosting agent and an immunostimulator for increasing the efficacy of FMDV vaccination in pigs.
Adjuvants, Immunologic/therapeutic use
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Aluminum Silicates/*therapeutic use
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Animals
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Antibodies, Viral/*immunology
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Antibody Formation/drug effects
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*Dietary Supplements
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Ferrous Compounds/*therapeutic use
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Foot-and-Mouth Disease/*immunology/prevention & control
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Foot-and-Mouth Disease Virus/immunology
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Germanium/*therapeutic use
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Swine
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Swine Diseases/immunology/prevention & control/*virology
6.Construction and immunization of an attenuated vaccine candidate enteropathogenic Escherichia coli O45 .
Yi HU ; Jie SONG ; Baohua ZHAO
Chinese Journal of Biotechnology 2009;25(2):181-188
In order to obtain an attenuated vaccine candidate for enteropathogenic Escherichia coli (EPEC) O45, a ler deletion mutant of pig enteropathogenic E. coli (PEPEC) O45 was constructed by using the suicide vector pCVD442, termed as PEPEC O45(deltaler). The culture supernatant of PEPEC O45(deltaler) deletion mutant was inoculated in vero cell culture. PEPEC O45(deltaler) deletion mutant lost the toxigenicity to vero cell. Test group and control group of mice were orogstrically inoculated with the PEPEC O45(deltaler) deletion mutant and the virulent strain O45 respectively. Mice were observed daily for clinical signs and weight changes. Test group of mice inoculated with PEPEC O45(deltaler) gained weight normally and experienced no clinical signs. In contrast, control group of mice inoculated with virulent strain O45 exhibited weight loss and all died in four days. In another experiment, pregnant mice and pig were orally vaccinated by PEPEC O45(deltaler) twice at interval of 14 days respectively. Subsequently, the suckling mice and pig were orally challenged with O45 at 7 days of age respectively. The results showed that 80% of the sucking mice born by vaccinated mice and 75% of the sucking pig born by vaccinated pig were survival; 15% of the sucking mice born by non-vaccinated mice and 10% of the sucking pig born by non-vaccinated pig were survival. This study demonstrated that PEPEC O45(deltaler) deletion mutant lost the toxigenicity to vero cell and to be safety to mice and pig. Oral immunization can induce specific immune responses in mice and pig, and this mutant strain could be used as an attenuated vaccine candidate against PEPEC O45.
Animals
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Enteropathogenic Escherichia coli
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genetics
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immunology
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Escherichia coli Infections
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microbiology
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prevention & control
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Escherichia coli Proteins
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genetics
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Escherichia coli Vaccines
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biosynthesis
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genetics
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immunology
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Gene Deletion
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Mice
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Mutagenesis, Site-Directed
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Swine
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microbiology
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Swine Diseases
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microbiology
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prevention & control
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Trans-Activators
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genetics
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Vaccines, Attenuated
;
biosynthesis
;
genetics
;
immunology
7.Prevalence of porcine proliferative enteropathy and its control with tylosin in Korea.
Sang Won LEE ; Tae Jong KIM ; Seung Yong PARK ; Chang Sun SONG ; Hyung Kwan CHANG ; Jae Kil YEH ; Hye In PARK ; Joong Bok LEE
Journal of Veterinary Science 2001;2(3):209-212
Porcine proliferative enteropathy(PPE) is an enteric disease been caused by Lawsonia intracellularis. It has become one of the critical problems in the pig industry. To investigate the prevalence of PPE in Korea, serum samples of 828 pigs from 65 herds were tested using indirect immunofluorescence antibody technique(IFA). The infection rate in individual pigs varied from 44 to 69%, whereas 100% in pig farms. The infection frequency was 57, 44.9, and 59.4% according to age respectively. Administration of tylosin in feed at a concentration of 110 ppm for 14 days reduced the infection rate of the farms. These data indicated that the high prevalence of PPE may be controlled by tylosin.
Administration, Oral
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Animal Feed
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Animals
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Anti-Bacterial Agents/*therapeutic use
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Antibodies, Bacterial/blood
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Enteritis/epidemiology/prevention&control/*veterinary
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Fluorescent Antibody Technique, Indirect/veterinary
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Gram-Negative Bacterial Infections/prevention&control/*veterinary
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Korea/epidemiology
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*Lawsonia Bacteria/drug effects/growth&development/immunology
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Seroepidemiologic Studies
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Swine
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Swine Diseases/*epidemiology/microbiology/prevention&control
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Tylosin/*therapeutic use