You are here
Determination and analysis of temperate phage genes proliferation in strains of Salmonella enterica
The occurrence of salmonellosis is growing every year which causes a great loss for agricultural industry. Mostly it’s referred to the poultry farming since the most common source of the infection is chicken meat, eggs and egg-containing products. Thereby there is urgency to find the ways to control effectively the epizootic context of salmonellosis.
The virulence factors in Salmonella are proteins, toxins and other specific substances which provide colonization, adhesion, invasion and mechanisms of resistance to phagocytosis in the host cells. The gens coding these traits are located within chromosomes of large plasmids. Routinely the virulence coding genes (virulence genes) interact with each other and are regulated mutually.
The acquirement of virulence genes can be caused by permanent circulation of Salmonella in the environment. One more source of the virulence genes acquirement is infection with the temperate bacteriophages. The result of that is entry the genes which can augment both invasion and virulence traits to the genome of bacteria. The temperate phages of Siphoviridae family (Gifsy-1, Gifsy-2) are commonly occurred in Salmonella. Gifsy-1 contribute the potential virulence gene, the most important of them is gipA. The expression of gipA determine the colonization of small inte stine, and its deletion leads to reduced virulence. The reason of that can be explained as the carrying gipA can give the ability to persist within Peyer’s patches. Similar effect on the bacterial virulence has the gene sodC1 of the prophage Gifsy-2. The product of his expression is Cu, Zn- superoxide dismutases. The are the virulence factor in salmonellas and can increase virulence fivefold. In addition there is strict mutuality between these two bacteriophages.
If Gifsy-2 is presented in the salmonella genome, Gifsy-1 does not show up. However Gifsy-2 can strengthen the pathogenicity of bacteria if Gifsy-2 is absent and gene sodC1 is integrated into the chromosome.
Thus genes of temperate phages can influence the virulence of salmonella. The research of their distribution within strains which circulate at poultry farms is important for finding the way to control epizootic situation of salmonellosis.
The main purpose of the present research was to detect the genes of temperate phages (sopE, sodC1, gipA) within strains of Salmonella enterica.
30 field strains of Salmonella genus obtained from several poultry farms in Ukraine have been researched. As well as 14 strains from National Center of the Strains (State Scientific Control Institute of Biotechnology and Strains of Microorganisms).
Lyophilized strains and the strains in the semi liquid agar were cultivated in the Nutrient broth at the 37˚ during 24 hours. After that all strains were streaked on the plates with XLD media (Xylose Lysine DEsoxycholate, Himedia, India).
In 24 hours 1–2 colonies were picked and used for DNA extraction using commercial kit “DNK-sorb-B” (Amplisense, Russia). The polymerase chain reaction was conducted using thermocyclers “Tercyk” (DNK-technologia, Russia) and “T1” (Biometra, Germany).
As a result the most prevalent gene of all three was sopE gene coding effector protein of T3SS1 which takes part in invasion mechanisms.
Gene GipA was found in 1 field strain (of 30 researched) and in 4 of 14 strains from the Collection of National Center of the Strains. The general percentage of gipA prevalence in the present research was 11 %.
Gene sopE was found in 70.5 % of the strains (26 of the 30 field strains and 5 of 14 strains from the Collection).
Gene sodC1 was found in 50 % of all strains (17 f the 30 field strains and 5 of 14 strains from the Collection).
The next step of our research must be the study of virulence of the strains carrying these 3 genes. And also the study of their lysogenic traits.
Key words: salmonella, genes, bacteriophages, factors pathogenicity, strains, poultry.
1. The European Union summary report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2014 / EFSA and ECDC (European Food Safety Authority and European Centre for Disease Prevention and Control) // EFSA Journal. – 2015. – Vol. 13, № 12 (4329). – P. 1–191.
2. The European Union Summary Report on Trends and Sources of Zoonoses, Zoonotic Agents and Food-borne Outbreaks in 2013 / EFSA and ECDC (European Food Safety Authority and European Centre for Disease Prevention and Control) // EFSA Journal. – 2015. – Vol. 13, № 1 (3991). – P. 1–165.
3. Salmonellosis // OIE. Terrestrial Manual. –2010. – Chapter 2.9.9. – P. 1–19.
4. The prevalence and PCR detection of Salmonella contamination in raw poultry / P. Whyte, K. Mc Gill, J.D. Collins, E. Gormley // Veterinary Microbiology. – 2002. – Vol. 89, № 1. – P. 53–60.
5. Sal'monelly: molekuljarnye mehanizmy prisposoblennosti i faktory virulentnosti / D.G. Ahmetova, Zh.A. Berdygulova, E.B. Evtyhova, A.V. Shustov // Biotehnologija. Teorija i praktika. – 2012. – № 1. – S. 3–24.
6. Ochman H. Lateral gene transfer and the nature of bacterial innovation / H. Ochman, J.G. Lawrence, E.A. Groisman // Nature. – 2000. –Vol. 405 (6784). – P. 299–304.
7. Shevchenko T.P. Virusy mikroorganizmiv / T.P. Shevchenko, I.G. Budzanivs'ka, V.P. Polishhuk // Kurs lekcij: navch. posibnyk. – K.: DP «Vydavnychyj dim «Personal», 2013. – 150 s.
8. Galan J.E. Protein delivery into eukaryotic cells by type III secretion machines / J.E. Galan, H. Wolf-Watz // Nature. – 2006. – Vol. 444 (7119). – P. 567–573.
9. Salmonella phages and prophages: genomics, taxonomy, and applied aspects. / A.I. Switt, A. Sulakvelidze, M. Wiedmann [et al.] // Methods Mol. Biol. – 2015. – Vol. 1225. – P. 237–287.
10. Bossi L. Prophage arsenal of Salmonella enterica serovar Typhimurium / L. Bossi, N. Figueroa-Bossi // Their Role in Bacterial Pathogenesis and Biotechnology / Eds.: M.K. Waldor, D.I. Friedman, S.L. Adhya. – Washington: ASM Press, 2005. – P. 165–186.
11. Stanley T.L. Tissue-specific gene expression identifies a gene in the lysogenic phage Gifsy-1 that affects Salmonella enterica serovar typhimurium survival in Peyer's patches / T.L. Stanley, C.D. Ellermeier, J.M. Slauch // Journal Bacteriology. – 2000. – Vol. 182, № 16. – P. 4406–4413.
12. Wagner P.L. Bacteriophage control of bacterial virulence / P.L. Wagner, M.K. Waldor // Infection and Immunity. – 2002. – Vol. 70, № 8. – P. 3985–3993.
13. Identification of GtgE, a Novel Virulence Factor Encoded on the Gifsy-2 Bacteriophage of Salmonella enterica Serovar Typhimurium / T.D. Ho, N. Figueroa-Bossi, M. Wang [et al.] // Journal Bacteriology. – 2002. – Vol. 184, № 19. – P. 5234–5239.
14. Figueroa‐Bossi N. Inducible prophages contribute to Salmonella virulence in mice. / N. Figueroa‐Bossi, L. Bossi // Molecular Microbiology. – 1999. – Vol. 33, № 1. – P. 167–176.
15. Isolation and characterization of Salmonella enterica in day-old ducklings in Egypt / Osman K.M., Marouf S.H., Zolnikov T.R., AlAtfeehy N. // Pathogens and Global Health. – 2014. – Vol. 108, № 1. – P. 37–48.
16. Detection of virulence-associated genes in Salmonella Enteritidis isolates from chicken in South of Brazil / K.A. Borges, T.Q. Furian, A. Borsoi [et al.] // Pesquisa Veterinária Brasileira. – 2013. – Vol. 33, № 12. – P. 1416–1422.
| Attachment | Size |
|---|---|
 2016_1_rublenko_n_ua.pdf | 815.91 KB |