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Number of results: 6
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Abstract

Highly immunogenic nucleotide fragments from 3 genes of Mycoplasma hyopneumoniae strain 232 were selected using information software technology. After repeating each fragment three times, a total of 9 nucleotide fragments were joined together to form a new nucleotide sequence called Mhp2321092bp. Mhp2321092bp was directly synthesized and cloned into a pET100 vector and expressed in Escherichia coli. After purification, the proteins were successfully validated by SDS-PAGE and Western blotting using mouse His-tag antibody and pig anti-Mhp serum. BALB/c mice were intraperitoneally injected with purified proteins in the high-dose (100 μg), medium-dose group (50 μg) and low-dose (10 μg) groups. Mice in each group were injected on day 1, day 8 and day 15 of feeding, respectively. Serum samples were collected from all mice on the day before immunization and on day 22 after immunization. The antibody level in the mouse serum was detected using western blotting using purified expressed proteins as antigens. IL-2, TNF-α and IFN-γ were simultaneously detected in the mouse serum by ELISA. The results showed that the 60 kDa protein was successfully expressed and reacted specifically with the specific serum Mhp His-Tag mouse monoclonal antibody and pig anti-Mhp serum. From day 0 to day 22 of immunization, IFN-γ increased from 269.52 to 467.74 pg/mL, IL-2 increased from 14.03 to 145.16 pg/mL, and TNF-α increased from 6.86 to 12.37 pg/mL. The IgG antibody in mice increased significantly from 0 day to day 22 after immunization. This study suggests that the expressed recombinant protein may serve as one of the novel vaccine candidates for Mhp.
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Authors and Affiliations

J. Li
1
G. Wang
1

  1. College of Veterinary Medicine, Hunan Agricultural University, Nongda road 1#, Changsha, Hunan, 410128, China
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Abstract

Staphylococcus aureus (S. aureus) has been recognized as one of the important zoonotic pathogens. However, it was limited about the epidemiology and genetic characteristics of S. aureus isolated from pigs in Hunan province, china. The aim of this study was to determine the characteristics of 163 S. aureus isolated from 590 pigs in Hunan Province, China. All isolates were characterized by agr typing, detection of virulence genes and antibiotic resistance genes, lethal test of mice and antibiotic susceptibility tests. The results showed that 30 strains of the 163 isolates were divided into agrI (18.40%), agrII (36/163, 22.09%), agrIII (20/163, 12.27%,), agrIV (20/163,12.27%) and the remaining 57 isolates were amplified negative by agr primers. In the 163 isolates, the detection rate of the virulence genes hlb, hld, hla, icaA, seb, fnbA, eta, etb, sea, tst and pvl ranged from 2.45% to 100%. The 43 isolates that were lethal to the mice, had β-hemolytic activity, the number of virulence genes of which was 7.8% higher than that of the remaining 120 non-fatal strains. The resistance rates of the 163 isolates to the 15 antibiotics were 0% (0/163) - 100% (163/163). All isolates were susceptible to Vancomycin and only 7 isolates were methicillin - resistant S. aureus (MRSA). The detection rates of the 11 resistance genes was 0% (0/163) - 100% (163/163). This study first to describes the epidemiology and characteristics of S. aureus from pigs in Hunan Province, which will help in tracking the evolution of epidemic strains and preventing pig-human transmission events.

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Authors and Affiliations

X. Zhang
G. Wang
C. Yin
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Abstract

A highly immunogenic nucleotide fragment (195bp) was selected from the Mhp183 gene of Mycoplasma hyopneumoniae using information software technology and was named Mhp183195bp. Three Mhp183195bp were linked to form a new nucleotide sequence called Mhp183615bp. Mhp183615bp was directly synthesized and cloned into a pET100 vector and expressed in Escherichia coli. After purification, the proteins were successfully validated using SDS-PAGE and Western blot. BALB/c mice were injected with purified proteins on the first, eighth, and fifteenth days of feeding, respectively; serum samples were collected from mice on the day of immunization and on the 22nd day after immunization. The antibody level in mouse serum was detected by Western blotting using purified expressed proteins as antigens. IL-2, TNF-α and IFN-γ were simultaneously detected in mouse serum by ELISA. The 30 kDa protein was successfully expressed and reacted specifically with the specific serum Mhp His-Tag mouse monoclonal antibody and pig antibody. The expressed recombinant protein was immunogenic. The expression levels of IFN-γ, IL-2 and TNF-α were found to be significantly higher on day 22 than in the control group. This study suggests that the expressed recombinant protein could be used as one of the novel vaccine candidates for Mhp.
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Bibliography


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Ferreira MR, Finger PF, Magalhes CG, Cunha CE, Junior CM, Kich JD, Mores N, Moreira AN, Dellagostin OA, Conceicao FR (2019) Protection efficacy of the rLTB-R1 chimera against experimental Swine Mycoplasmal Pneumonia. Acta Sci Vet 47: 1660.

Huo SX, Zeng ZZ, Zhao YL, Han RL (2019) Effect of antigen purification on the efficacy and safety of inactivated swine Mycoplasma hyopneumoniae vaccine. Anim Husb Vet Med 51: 87-92.

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Wang Y, Wang J, Zhou M, Liu P, Zhang E, Li Y, Feng Z, Yang Q (2019) Mucosal and systemic immune responses induced by intranasal immunization of recombinant Bacillus subtilis expressing the P97R1, P46 antigens of Mycoplasma hyopneumoniae. Biosci Rep 39: BSR20191126.

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Authors and Affiliations

M. Xu
1
J. Zheng
1
S. Hu
1
G. Wang
1

  1. College of Veterinary Medicine, Hunan Agricultural University, Road 1#, Changsha, 410000, China
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Abstract

The main purpose of the study is to investigate the mechanical properties around an underground gas storage cavern in bedded salt rock. Firstly, considering the characteristics of the salt rock formation in China, the mechanical model was simplified into a hollow cylinder, which containing non-salt interlayer. In terms of elastic theory, Love displacement function was developed, and the elastic general solution of stress and deformation components were obtained after determining the undetermined coefficients. Under the same condition, numerical simulation was carried out. The validity of the elastic general solution is verified by comparing to numerical simulation results. Furthermore, Based on the feasible general elastic solution, viscoelastic solution was obtained through Laplace transformation and inverse Laplace transform, which could provide reference for the study on the stability and tightness of underground gas storage carven during operation to some extent.

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Authors and Affiliations

P. Xie
H.J. Wen
G.J. Wang
J. Hu
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Abstract

Classical swine fever (CSF) and porcine reproductive and respiratory syndrome (PRRS) are responsible for major economic losses and represent a threat to the swine industry worldwide. Routine surveillance serology for CSF and PRRS viruses is critical to maintaining the health status of sow farms in Hunan Province, which is one of the top pig production provinces in China. The aim of our study was to investigate the serological statistics of CSF virus (CSFV) and PRRS virus (PRRSV) in Hunan Province. The cohort serum samples were collected from vaccinated and unvaccinated pigs. Our findings showed that the average rates of CSFV and PRRSV antibody seropositivity were 82.2% (95% CI: 80.1-84.3) and 84.8% (95% CI: 82.5-87.1), respectively, in the immunized group and that these rates were higher than those in the unvaccinated group (58.6% for CSFV and 47.8% for PRRSV). Additionally, the level of CSFV antibody in piglet serum declined gradually with age, whereas PRRSV-specific antibody level increased initially (1 to 2 weeks old) and then declined with age (2 to 4 weeks old). In summary, we investigated the difference in CSFV/PRRSV antibody levels among piglets at various weeks old (1 to 4 weeks) to further establish the duration of maternal immunity in piglets. In addition, routine monitoring of CSFV/PRRSV antibodies in immunized pigs was carried out to evaluate the efficacy of vaccination.
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Bibliography


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Authors and Affiliations

H. Yu
1
L. Zhang
1
Y. Cai
1
Z. Hao
2
Z. Luo
3
T. Peng
1
L. Liu
N. Wang
1
G. Wang
1
Z. Deng
1
Y. Zhan
1

  1. Provincial Key Laboratory of Protein Engineering in Animal Vaccines, Research Center of Reverse Vaccinology (RCRV), and Laboratory of Functional Proteomics (LFP), College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan 410128, China
  2. Yongzhou Animal Husbandry and Aquatic Affairs Center, Yongzhou, Hunan 425000, China
  3. Dingcheng Animal Husbandry and Aquatic Affairs Center, Changde, Hunan 415100, China

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