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Preliminary studies on some avian diseases in Antarctic birds

Gerardo Leotta Raul Cerda Nestor R. Coria Diego Montalti
Polish Polar Research | 2001 | vol. 22 | No 3-4 | 227-231
Keywords Antarctica birds Mycoplasma Salmonella
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Abstract

A serological study to detect antibodies against microbes in avian mycoplasmosis (Mycoplasma gallisepticum and M. synoviae), and salmonellosis (Salmonella gallinarum and S. pullorum) was carried out. A hundred and twelve Antarctic birds (42 Adelie penguins, Pygoscelis adeliae, 30 southern giant petrels, Macronectes giganteus and 40 skuas, Catharacta antarctica and C. maccormicki) from King George Island, the South Shetland Islands, and Laurie Island, the South Orkney Islands in Antarctica were studied. The serological test used in this study was a rapid agglutination test. According to the results and considering the number of samples analysed, it is reasonable to believe that Adelie penguins, southern giant petrels, and skuas populations of the areas mentioned above are free from mycoplasmosis and salmonellosis.

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Authors and Affiliations

Gerardo Leotta
Raul Cerda
Nestor R. Coria
Diego Montalti

A new strategy: high level expression and immunogenicity analysis of triplicate repeated multigenes in Mycoplasma hyopneumoniae

J. Li G. Wang
Polish Journal of Veterinary Sciences | 2023 | vol. 26 | No 2 | 275-283 | DOI: 10.24425/pjvs.2023.145034
Keywords multi-epitope fusion protein Mycoplasma hyopneumoniae new generation vaccines
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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

Expression and immunogenicity study of a novel mhp183 gene fragment of Mycoplasma hyopneumoniae

M. Xu J. Zheng S. Hu G. Wang
Polish Journal of Veterinary Sciences | 2021 | vol. 24 | No 4 | 553-561 | DOI: 10.24425/pjvs.2021.139980
Keywords Mycoplasma hyopneumoniae Mhp183 gene new generation vaccines
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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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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

Prevalence, molecular characterization and antibiogram of Mycoplasma bovis isolated from milk in Pakistan

A. Jabbar M. Ashraf S.U. Rahman M.S. Sajid
Polish Journal of Veterinary Sciences | 2023 | vol. 26 | No 3 | 461–471 | DOI: 10.24425/pjvs.2023.145054
Keywords milk Mycoplasma bovis modified hayflick medium PCR DNA sequencing disc diffusion antibiotic susceptibility
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Abstract

Mycoplasma bovis is a highly contagious pathogen that causes clinical or subclinical mastitis. The present study was aimed for the isolation, molecular characterization and antibiogram determination of M. bovis from raw milk samples. Milk samples were collected randomly from lactating cows and buffaloes from different tehsils of district Faisalabad, Pakistan. Samples were inoculated on modified Hayflick medium and biochemical tests were performed for further confirmation of isolated M. bovis. Out of total 400 milk samples, 184 (46%) samples were found positive for culture method. The 16S-rRNA gene polymerase chain reaction was performed for molecular characterization of isolated M. bovis strains. Out of total 400 milk samples, 240 (60%) positive for M. bovis through PCR method were examined. The 16S-rRNA gene PCR positive isolated M. bovis strains were sequenced and results were compared using Maximum-likelihood method and sequenced strains of M. bovis were aligned and analyzed by Clustal W software. Antibiogram of isolated M. bovis strains was analyzed by disc diffusion assay against eight commonly used antibiotics. Tylosin (30μg) and Tilmicosin (15ug) showed inhibition zones of 32.34 ± 1.10 mm and 17.12 ± 0.93 mm respectively against isolated M. bovis which were found sensitive. Isolated M. bovis was found resistant to other commonly used antibiotics. Statistical analysis revealed that p-value was < 0.05 and the odds ratio was >1.0 at 95% CI. This study complemented the lack of epidemiological knowledge of molecular characterization, comparative effectiveness and resistance trends of isolated M. bovis strains against commonly used antibiotics.
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Authors and Affiliations

A. Jabbar
1
M. Ashraf
1
S.U. Rahman
1
M.S. Sajid
2
  1. Institute of Microbiology, University of Agriculture, Jail Road, Faisalabad, Punjab 38000, Pakistan
  2. Department of parasitology, University of Agriculture, Jail Road, Faisalabad, Punjab 38000, Pakistan

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