Detection of virulence factors of Escherichia coli strains isolated from urogenital system infections in dogs and cats

Year 2021, , 132 - 142, 15.06.2021

Orkun Babacan , Müjgan İzgür

https://doi.org/10.33188/vetheder.839260

Abstract

In this study, virulence factors such as serum resistance, aerobactin iron uptake systems, adhesins (type 1 fimbria, P fimbria, S fimbria, afimbrial adhesins), haemolysine, uropathogenic spesific protein and cytotoxic necrotizing factor in 45 Escherichia coli isolates from dogs and cats with urinary tract and genital system infections, were investigated by both phenotypic methods and molecular methods. In PCR examinations of 45 E. coli, hlyA gene was detected in 73.3% of all isolates. fimH was found in 100%, papC in 71.1%, sfaDE in 82.2%, and afaBC in 0% of the isolates. iucD, traT, usp, cnf1 and cvaC genes were found in 20%, 71.1%, 84.4%, 75.5%, and 11.1% of the isolates, respectively. According to the isolation rate of hlyA and haemolytic activity on blood agar, this virulence factor is commonly seen in these strains and important in these infections especially pathogenesis. İsolates was found to be a high rate positive for usp gene. This result suggested that this strain may have higher infectivity. Cytotoxic necrotizing factor was common and important virulence factor in uropathogenic E. coli and prognosis of infection. However, colisin encoding gene (cvaC) only were detected from 5 of urine samples from dogs. This virulence factor could be important for the urinary tract infections in dogs rather than cats. Haemolysis, type 1 fimbriae, P fimbriae, S fimbriae, cnf, serum resistance, uropathogenic specific protein were detected at high rates by molecular method. In E. coli strains isolated from dogs, virulence factors were found to be higher than cat isolates.

Keywords

cat, dog, esherichia coli, urogenital system infections, virulence genes

Kedi ve köpeklerin ürogenital sistem infeksiyonlarından izole edilen Escherichia coli'lerin virulens faktörlerinin belirlenmesi

Abstract

Bu çalışmada, köpek ve kedilerin idrar yolu ve genital sistem enfeksiyonlarından izole edilen 45 Escherichia coli izolatlarında serum direnci, aerobaktin demir alım sistemleri, adezinler (tip 1 fimbria, P fimbria, S fimbria, afimbrial adezinler), hemolizin, üropatojenik spesifik protein ve sitotoksik nekrotizan faktör gibi virülans faktörleri hem fenotipik yöntemler hem de moleküler yöntemler ile araştırıldı. 45 E. coli izolatının virulens genlerine ait PCR sonuçlarında, tüm izolatların % 73.3'ünde hlyA geni tespit edildi. İzolatların% 100'ünde fimH,% 71.1'inde papC, % 82.2'sinde sfaDE ve % 0'ında afaBC bulundu. iucD, traT, usp, cnf1 ve cvaC genleri, izolatların sırasıyla% 20,% 71.1,% 84.4,% 75.5 ve % 11.1'inde bulundu. HlyA'nın izolasyon oranı ve kanlı agarındaki hemolitik aktivitesine göre, bu virülans faktörü bu suşlarda yaygın olarak görüldüğü ve bu enfeksiyonlarda özellikle patogenezde önemli olduğu düşünüldü. İzolatlarda usp geni için yüksek oranda pozitif olduğu bulunmuştur. Bu sonuç, bu bakterilerin daha yüksek bir enfektiviteye sahip olabileceğini düşündürmektedir. Sitotoksik nekrotizan faktör üropatojenik E. coli ve enfeksiyon prognozu için yaygın ve önemli virülans faktörüdür. Bununla birlikte, köpeklerden idrar numunelerinin sadece 5'inde kolisin kodlayan gen (cvaC) tespit edildi. Bu virülans faktörü, kediler yerine köpeklerde idrar yolu enfeksiyonları için önemli olabilir. Hemoliz, tip 1 fimbria, P fimbria, S fimbria, cnf, serum direnci, üropatojenik spesifik protein moleküler yöntemle yüksek oranlarda saptandı. Köpeklerden izole edilen E. coli suşlarında, virülans faktörlerinin kedi izolatlarından daha yüksek olduğu bulundu.

Keywords

Escherichia coli, kedi, köpek, ürogenital sistem infeksiyonları, virulens genleri

References

• 1. Alonso P, Blanco J, Blanco M, Gonzalez EA (1987): Frequent production of toxins by Escherichia coli strains isolated from human urinary tract infections: relation with haemagglutination. FEMS Microbiol Lett, 48, 391- 396.
• 2. Arnberg, J, Flagstad A (1985): Prostaglangin F2α Treatment of Feline Pyometra. Nord Vet Med, 37, 286- 290.
• 3. Arthur M, Johnson CE, Rubin RH, Arbeit RD, Campanelli C, Kim C, Stainbach S, Agarwal M, Wilkinson R, Goldstain R (1989): Molecular epidemiology of adhesin and hemolysin virulence factors among uropathogenic Escherichia coli. Infect Immun, 52, 303- 313.
• 4. Bağcigil AF, Dokuzeylül B, Göçmen H, Yalçin E, Ikiz S, Özgür NY, Ak S (2018): The characterization of Escherichia coli strains isolated from urinary tract infections in cats and dogs. Anatolian Env and Anim Sciences, 3 (3), 131-136.
• 5. Bauer RJ, Zhang L, Foxman B, Siitonen A, Jantunen ME, Saxen H, Marrs CF (2002): Molecular epidemiology of 3 putative virulence genes for Escherichia coli urinary tract ınfection-usp, iha, and iroN. J Infect Dis, 185, 1521-1524.
• 6. Beutin L (1999): Escherichia coli as a pathogen in dogs and cats. Vet Res, 30, 285- 289.
• 7. Birosowa E, Siegfried E, Kmet’ova M, Makara A, Ostro A, Gresova A, Urdzik P, Liptakova A, Molokacova M, Bartl R, Valansky L (2004): Detection of virulence factors in a-haemolytic Escherichia coli strains isolated from various clinical materials. Clin Microbiol Infect, 10, 569- 573.
• 8. Cantekin Z, İzgür M (2015): Detection of virulence properties escherichia coli originated from poultry by phenotypic and molecular methods. Van Vet J, 26 (2), 71-76.
• 9. Dinç G (2009): Determination of virulence factors of Escherichia coli isolated from cows with mastitis and enviromental sources. PhD Thesis, Ankara University Health Science Institute, Ankara, Turkey.
• 10. Etefia EU, Ben SA (2020): Virulence markers, phylogenetic evolution, and molecular techniques of uropathogenic Escherichia coli. J Nat Sci Med, 3,13-22.
• 11. Fidan I, Yüksel S, Sipahi AB, Özkan S, Sultan N (2006): Haemagglutination and Haemolysin Production of Esherichia coli Isolates from Urinary Tract Infections. ANKEM Derg, 20, 22- 25.
• 12. Fritag T, Squires RA, Schmid J, Elliot J (2005): Feline uropathogenic Escherichia coli from Great Britain and New Zealand have dissimilar virulence factor genotypes. Vet Microbiol, 106, 79- 86. 13. Gatoria IS, Saini S, Rai TS, Dwivedi PN (2006): Comparison of Three Techniques for the Diagnosis of Urinary Tract Infections in Dogs with Urolithiasis. J Small Anim Pract, 47, 727- 732.
• 14. Gürdal H, (2003): Comparison of Bacteria Virulents Factors as a Reason of Urinary Infection and Sociocultural Levels of Patients. PhD Thesis, Süleyman Demirel University, Isparta, Turkey.
• 15. Hagman G, Greko C (2005): Antimicrobial resistance in Escherichia coli isolated from bitches with pyometra and from urine samples from other dogs. Vet Rec, 157, 193-197.
• 16. Hagman R (2004): New aspects of canine pyometra: studies on epidemiology and pathogenesis. Doctoral Thesis, Sweedish University of Agricultural Sciensis, Uppsala, Sweeden.
• 17. Johnson JR, Kaster N, Kuskowski MA, Ling GV (2003): Identification of urovirulence traits in Escherichia coli by comparison of urinary and rectal E. coli ısolates from dogs with urinary tract ınfection. J Clin Microbiol, 41, 337- 345.
• 18. Johnson JR, Stell AL (2000): Extended virulence genotypes of Escherichia coli strains from patients with urosepsis in relation to phylogeny and host compromise. J Infect Dis, 181, 261- 272.
• 19. Johnson JR (1991): Virulence factors in Escherichia coli urinary tract infection. Clin Microbiol Rev, 4, 80- 128.
• 20. Kanamaru S, Kurazono H, Nakano M, Terai A, Ogawa O, Yamamoto S (2006): Subtyping of uropathogenic Escherichia coli according to the pathogenicity island encoding uropathogenic-specific protein: Comparison with phylogenetic groups. Int J Urol, 13, 754- 760.
• 21. Kurazono H, Yamamoto S, Nakano M, Nair GB, Terai A, Chaicumpa W, Hayashi H (2000): Characterization of a putative virulence island in the chromosome of uropathogenic Escherichia coli possessing a gene encoding a uropathogenic-specific protein. Microb Pathog, 28, 183-189.
• 22. Litster A, Moss SM, Plettel J, Trott DJ (2009): Occult Bacterial Lower Urinary Tract Infections in Cats—Urinalysis and Culture Findings. Vet Microbiol, 136, 130- 134.
• 23. Marrs CF, Zhang L, Foxman B (2005): Escherichia coli mediated urinary tract infections: Are there distinc uropathogenic E. coli (UPEC) pathotypes? FEMS Microbiol Lett, 252, 183- 190.
• 24. Moyaert H, Morrissey I, de Jong A, El Garch F, Klein U, Ludwig C, Thiry J, Youala M (2017): Antimicrobial susceptibility monitoring of bacterial pathogens isolated from urinary tract infections in dogs and cats across europe: compath results. Microbial Drug Res, 23 (3), 391-403.
• 25. Mustapha M, Goel P (2020): Isolation and prevalence of uropathogenic E. coli in different breed, sex and age of dogs. Rev Vét Clin, 142. Doi: https://doi.org/10.1016/j.anicom.2020.02.004
• 26. Osman KM, Hessain AM, Abo-shama UH, Girh ZM, Kabli SA, Herneg HA, Moussa IM (2018): An alternative approach for evaluating the phenotypic virulence factors of pathogenic Escherichia coli. Saudi Biomed Sci, 25, 195-197.
• 27. Papini R, Ebani VV, Cerri D, Guidi G (2006): Survey on bacterial isolates from dogs with urinary tract infections and their in vitro sensitivity. Rev Méd Vét, 157, 35- 45.
• 28. Prssler B, Bartges J (2010): Urinary tract infections. 2082- 2093. In: Ettinger SJ, Feldman EC (Eds.) Textbook of Veterinary Internal Medicine, Elsevier Saunders, St. Louis, MO, USA.
• 29. Quinn PJ, Markey BK, Carter ME, Donnely MJ, Leonard FC (2002): Veterinary Microbiology and Microbiol Disease, Replika Pres Pvt. Ltd, India.
• 30. Raksha R, Srinivasa H, Macaden RS (2003): Occurrance and characterisation of uropathogenic Escherichia coli in urinary tract infections. Indian J Med Microbiol, 21, 102- 107.
• 31. Rijavec M, Muller‑Premru M, Zakotnik B, Zgur‑Bertok D (2008): Virulence factors and biofilm production among Escherichia coli strains causing bacteraemia of urinary tract origin. J Med Microbiol, 57, 1329‑34.
• 32. Ronald A (2002): The etiology of urinary tract ınfection: traditional and emerging pathogens. Am J Med, 49 (2), 71-82.
• 33. Sanchez- Carlo V, Mcdonald JS, Packer RA (1984): Virulence factors of Escherichia coli isolated from cows with acute mastitis. Am J Vet Res, 45, 1775- 1777.
• 34. Siqueira AK, Riberio AG, Leite DS, Tiba MR, Moura CD, Lopes MD, Prestes NC, Salerno T, Silva AV (2009): Virulence factors in Escherichia coli strains isolated from urinary tract infection and pyometra cases and from feces of healthy dogs. Res Vet Sci, 86, 206- 210.
• 35. Stuart ST, Grenwod KT, Luke RKJ (1980): Hidroxamate-mediated transport of iron controlled by Col V plasmid. J Bacteriol, 143, 35- 42.
• 36. Sussman M, Gally DL (1999): The Biology of Cystitis: Host and Bacterial Factors. Annu Rev Med, 50, 149- 158.
• 37. Svanborg C, Bergsten G, Fischer H, Godaly G, Gustafsson M, Karpman D, Lundstedt AC, Ragnarsdottir B, Svensson M, Wullt B (2006): Uropathogenic Escherichia coli as a model of host- parasite interaction. Curr Opin Microbiol, 9, 33- 39.
• 38. Wadas B, Kuhn I, Lagerstedt AS, Jonsson P (1996): Biochemical phenotypes of Escherichia coli in dogs: Comparison of isolates isolated from bitches sufkring from pyometra and urinary tract infection with isolates from faeces of healthy dogs. Vet Microbiol, 52, 293- 300.
• 39. Wells JE, Bartges JW, Kania SA, Bemis DA, Gluhak T (2013): Association between Presence of Urovirulence Factors, Phylogenetic Class, and Antimicrobial Resistance Patterns in 159 Uropathogenic Escherichia coli Samples Isolated from Dogs. Open J Vet Med, 3, 199-203.
• 40. Yamamoto S, Nakano M, Terai A, Yuri K, Nakata K, Nair GB, Kurazono H, Ogawa O (2001): The Precence of The Virulence Island Containing the usp Gene in Uropathogenic Escherichia coli is Associated with Urinary Tract Infection in An Experimental Mouse Model. J Urol, 165, 1347- 1351.
• 41. Yamamoto S, Terai A, Yuri K, Kurazono H, Takeda Y, Yoshida O (1995): Detection of Urovirulence Factors in Escherichia coli by Mutltiplex Polymerase Chain Reaction. FEMS Immunol Med Microbiol, 12, 85-90.