Comparative study on genotypic properties of Pseudomonas aeruginosa isolated from subclinical mastitis in Türkiye

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Published: Apr 18, 2023
Updated: 2023-04-18
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2023-04-18 (2)
DOI: https://doi.org/10.12681/jhvms.29590
Keywords:
antibiotic resistance cow milk integron gene P. aeroginosa virulence gene
S Turkyilmaz
Aydın Adnan Menderes Üniversitesi
https://orcid.org/0000-0002-1363-4534
F Ocak
Abstract

The results of antibacterial therapy in mastitis may be related to not only the antibiotic sensitivity of the etiological factors, but also to their abilities carrying integron and virulence genes. Integrons are mobile DNA element that can capture and carry genes, particularly those responsible for antibiotic resistance. Pseudomonas aeroginosa has numberless virulence factors which are contributed to bacterial invasion and toxicity. The aim of this study is to investigate antibiotic resistance, integron and virulence gene profiles of P. aeruginosa isolates obtained from cow milk with subclinical mastitis; further, was to evaluate the relationship between the antimicrobial resistance of bacteria with the of integron and virulence gene carrying. Material of the study is consisted with 32 (9.8%) of P. aeruginosa isolates obtained from 326 subclinical mastitic milk samples. After the bacterial identification was done by classical conventional methods, polymerase chain reaction (PCR) was used to verify the genus and species identifications of the isolates and to determine the integron and virulence gene profiles. Using disk diffusion method, antimicrobial resistance was determined to fifteen antibiotics against belonging to eleven antimicrobial families. The relationship the presence of integron and virulence genes associated with antibiotic resistance, further the relationship the presence of virulence genes and antimicrobial resistance was calculated with the Chi-Square (χ2) test. Ten of virulence genes (lasl, lasR, lasB, rhll, rhlR, rhlAB, plcH, plcN, ppyR, exoT) were found in all isolates whilst a virulence gene (aprA) was not present in any isolate. It was found that 34.4% of the isolates carried any integron gene. The results showed that the relationship is important between the presence of int genes and gentamicin, amikacin, tetracycline, cefoperazone, imipenem, aztreonam, ciprofloxacin, enrofloxacin resistance and exoU virulence gene presence. Also; there were also important associations between resistance to certain antibiotics and the presence of P. aeroginosa virulence genes: ciprofloxacin with lasA; netilmicin, cefoperazone and ciprofloxacin with pilB; netilmicin and ciprofloxacin with pelA; amikacin with algD, algU, algL; aztreonam with toxA; enrofloxacin with pvdA. All isolates obtained in this study showed multiple antibiotic resistance (MDR). In these cases, the presence of integron and some virulence genes could play a prominent role in the resistance of P. aeroginosa isolates to antimicrobial drugs. This study could be important in that it is the first study to show the presence of integrons and virulence genes in P. aeroginosa isolates originating from mastitic cow milk in Turkey. The presence of antibiotic-resistant P. aeroginosa strains in cattle farms may also pose a public health risk, as these bacteria can transmit their resistance genes to humans through food consumption.

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References
Aggarwal S (2008) Techniques in Molecular Biology. Lucknow: International Book Distributing CO. Short tandem repeat genotyping; pp.: 127-134.
Ahmed AM and Shimamoto T (2011) Molecular characterization of antimicrobial resistance in Gram-negative bacteria isolated from bovine mastitis in Egypt. Microbiol. Immunol. 55: 318–327.
Ajayi T, Allmond LR, Sawa T and Wiener-Kronish JP (2003) Single-nucleotide-polymorphism mapping of the Pseudomonas aeruginosa type III secretion toxins for development of a diagnostic multiplex PCR system. J. Clin. Microbiol. 41 (8): 3526-3531.
Ama A, El-Shafii SS, Amo AE and El-dayim ZAA (2016) Be detection of multidrug resistance genes in Pseudomonas aeruginosa isolated from bovine mastitic milk. JDVAR 3(2): 43-49.
Banerjee S, Batabyal K, Joardar SN, Isore DP, Dey S, Samanta I and Samanta TK (2017) Mice pathology study of toxA and exoS genes bearing Pseudomonas aeruginosa isolated from bovine sub-clinical mastitis in West Begal with their Antibiogram. Indian J. Anim. Res. 52(8): 1-6.
Bass L, Liebert CA, Lee MD, Summers AO, White DG, Thayer SG and Maurer JJ (1999) Incidence and characterization of integrons, genetic elements mediating multiple-drug resistance, in avian Escherichia coli. Antimicrob. Agents Chemother. 43: 2925–2929.
Chengappa MM (1990) Antimicrobial Agents and Susceptibility Testing. In Carter GR, Cole JR. (Eds.) Diagnostic Procedures in Veterinary Bacteriology and Mycology. 5th edition, Academic Press Inc., San Diego, California.
CLSI (2012) Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Second Informational Supplement. CLSI document M100-S22. Wayne, PA: Clinical and Laboratory Standards Institute.
Denning GM and Iyer SS, Reszka KJ, O’Malley Y, Rasmussen GT and Britigan BE (2003) Phenazine-1-carboxylic acid, a secondary metabolite of Pseudomonas aeruginosa, alters expression of immunomodulatory proteins by human airway epithelial cells. Am. J. Physiol. Lung Cell. Mol. Physiol. 285: L584–L592.
Engel J and Balachandran P (2009) Role of Pseudomonas aeruginosa type III effectors in disease. Curr. Opin. Microbiol. 12: 61–66.
Ertugrul BM, Oryasin E, Lipsky BA, Willke A and Bozdogan B (2018) Virulence genes fliC, toxA and phzS are common among Pseudomonas aeruginosa isolates from diabetic foot infections. Infect. Dis. 50 (4): 273–279.
Faghri J, Nouri S, Jalalifar S, Zalipoor M and Halaji M (2018) Investigation of antimicrobial susceptibility, class I and II integrons among Pseudomonas aeruginosa isolates from hospitalized patients in Isfahan, Iran. BMC Res. Notes 11: 806.
Fazeli N and Momtaz H (2014) Virulence gene profiles of multidrug-resistant Pseudomonas aeruginosa isolated from Iranian hospital infections. Iran Red. Crescent Med. J. 16 (10): e15722.2.
Fleiszig S, Wiener-Kronish JP, Miyazaki H, Vallas V, Mostov KE, Kanada D, Sawa T, Yen T and Frank DW (1997) Pseudomonas aeruginosa-mediated cytotoxicity and invasion correlate with distinct genotypes at the loci encoding exoenzyme S. Infect. Immun. 65, 579–586.
Fouad Z (2011) Antimicrobial Disk Diffusion Zone Interpretation Guide. https://www.researchgate.net/publication/315844044. DOI: 10.13140/RG.2.2.13801.70240.
Gasink LB, Fishman NO, Weiner MG, Nachamkin I and Bilker WB (2006) Fluoroquinolone-resistant Pseudomonas aeruginosa: assessment of risk factors and clinical impact. Am. J. Med. 119(6): 526e19-25.
Ghadaksaz A, Fooladi AAI, Hosseini HM and Amin M (2015) The prevalence of some Pseudomonas virulence genes related to biofilm formation and alginate production among clinical isolates. J. Appl. Biomed. 13: 61-68.
Goldstein C, Lee MD, Sanchez S, Hudson C, Phillips B, Register B, Grady M, Liebert, C, Summers AO, White DG and Maurer JJ (2001) Incidence of class 1 and 2 integrases in clinical and commensal bacteria from livestock, companion animals, and exotics. Antimicrob. Agents Chemother. 45: 723-726.
Goli HR, Nahaei MR, Rezaee MA, Hasani A, Kafl HS and Aghazadeh, M (2017) Prevalence and molecular characterization of class 1 integrons among clinical isolates of Pseudomonas aeruginosa in Northwest of Iran. Mol. Gen. Microbiol. Virol. 32:109–11.
Greenberg EP (1997) Quorum sensing in Gram-negative bacteria. ASM News 63: 371-377.
Hentzer M, Teitzel GM, Balzer GJ, Heydorn A, Molin S, Givskov M and Parsek MR (2001) Alginate overproduction affects Pseudomonas aeruginosa biofilm structure and function. J. Bacteriol. 183: 5395–5401.
Howe TR and Iglewski BH (1984) Isolation and characterization of alkaline protease-deficient mutants of Pseudomonas aeruginosa in vitro and in a mouse eye model. Infect. Immun. 43: 1058-1063.
Kipnis E, Sawa T and Wiener-Kronish J (2006) Targeting mechanisms of Pseudomonas aeruginosa pathogenesis. Med. Mal. Infect. 36:78-91.
Kirk JH and Bartlett PC Nonclinical Pseudomonas aeruginosa mastitis in a dairy herd. J. Am. Vet. Med. Assoc. 184: 671–673, 1984.
Kouchaksaraei FM, Shahandashti EF, Molana Z, Kouchaksaraei MM, Asgharpour F, Mojtahedi A and Rajabnia R (2012) Molecular detection of integron genes and pattern of antibiotic resistance in Pseudomonas aeruginosa strains isolated from intensive care unit, Shahid Beheshti Hospital, North of Iran. IJMCM 1(4): 2009-2016.
Magiorakos AP, Srinivasan A, Carey RB, Carmeli Y, Falagas ME, Giske CG, Harbart S, Hindler JF, Kahlmeter G, Olsson-Liljequist B, Paterson DL, Rice LB, Stelling J, Struelens MJ, Vatopoulos A, Weber JT and Monnet DL (2012) Multidrug-resistant, extensively drug resistant and pandrug resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin. Microbiol. Infect. 18: 268–281.
Mazel D (2006) Integrons: agents of bacterial evolution. Nat. Rev. Microbiol. 4:608–620.
McIntyre-Smith A, Schneiderman J and Zhou K (2010) Alginate does not appear to be essential for biofilm production by PAO1 Pseudomonas aeruginosa. JEMI 14: 63–68..
Meyer JM, Neely A, Stintzi A, Georges C and Holder IA (1996) Pyoverdin is essential for virulence of Pseudomonas aeruginosa. Infect. Immun. 64: 518–523.
Mokhtari AR, Salehi ZT, Amini K and Shahcheraghi F (2016) Isolation Pseudomonas aeroginosa bacteria and genes integron class 1 of subclinical mastitis in dairy cows in Tehran. Veterinary Researches Biological Products 29(2): 37-44.
Neamah AA (2017) Molecular detection of virulence factor genes in Pseudomonas aeruginosa isolated from human and animals in Diwaniya province. Kufa J. Vet. Med. Sci. 8(1): 218-230.
Ozturk D, Sahan Yapıcıer O, Sababoglu E, Kaya M, Pehlivanoglu F and Turutoglu H (2019) Antibiotic resistance of Gram negative bacteria isolated from bovine mastitis. Van Vet. J. 30 (2): 85-89.
Park SJ, Kim AK, So YI, Park HY, Li XH, Yeom DH, Lee MN, Lee BL and Lee JH (2014) Protease IV, a quorum sensing-dependent protease of Pseudomonas aeruginosa modulates insect innate immunity. Mol. Microbiol. 94 (6): 1298-1314.
Quinn PJ, Markey BK, Leonard FC, FitzPatrick ES, Fanning S and Hartigan PJ (2011) Veterinary Microbiology and Microbial Disease. Second Edition, Blackwell Science Ltd, Oxford, UK.
Roy-Burman A, Savel RH, Racine S, Swanson BL, Revadigar NS, Fujimoto J, Sawa T, Frank DW and Wiener-Kronish JP (2001) Type III protein secretion is associated with death in lower respiratory and systemic Pseudomonas aeruginosa infections. J. Infect. Dis. 183(12): 1767–1774.
Sabharwal N, Dhall S, Chhibber S and Harjai K (2014) Moleculer detection of virulence genes as markers in Pseudomonas aeruginosa isolated from urinary tract infections. IJMEG 5 (3): 125-134.
Sahin C and Erbas G (2015) Antibiotic susceptibilities and isolation of Pseudomonas aeruginosa from clinical mastitis in cattle. Etlik J. Vet. Microbiol. 26 (1): 16-20.
Selezska L, Wagner G, Gramer N, Siebert B, Gudowius P, Morales G, Kohler T, Van Delden C, Weinel C and Slickers P (2007) Population structure of Pseudomonas aeruginosa. PNAS USA, 104: 8101–8106.
Spilker T, Coenye T, Vandamme P and LiPuma JJ (2004) PCR-Based for differentiation of Pseudomonas aeruginosa from other Pseudomonas species recovered from cystik fibrosis patients. J. Clin. Microbiol. 42 (5): 2074-2079.
Szmolka A, Cramer N and Nagy B (2012) Comparative genomic analysis of bovine, environmental and human strains of Pseudomonas aeruginosa. FEMS Microbiol. Lett. 335: 113–122.
Tel OY, Keskin O, Zonturlu AK and Arserim KNB (2009) Subclinical mastitis prevalance and determination of the antibiotics susceptibility in Sanliurfa Region. Fırat Üni. J. Health Sci. (Vet) 23 (2): 101-106.
Vance RE, Rietsch A and Mekalanos JJ (2005) Role of the type III secreted exoenzymes S, T, and Y in systemic spread of Pseudomonas aeruginosa PAO1 in vivo. Infect. Immun. 1706–1713.
Wiehlmann L, Wagner G, Gramer N, Siebert B, Gudowius P, Morales G, Kohler T, Van Delden C, Weinel C and Slickers P (2007) Population structure of Pseudomonas aeruginosa. PNAS USA, 104: 8101–6.
Wiener-Kronish JP, Sakuma T, Kudoh I, Pittet JF, Frank D, Dobbs L, Vasil ML, and Matthay MA (1993) Alveolar epithelial injury and pleural empyema in acute P. aeruginosa pneumonia in anesthetized rabbits. J. Appl. Physiol. 75(4): 1661–1669.
Woese CR (1987) Bacterial evolution. Microbiol. Rev. 51: 221–271,.
Woods DE and Iglewski BH (1983) Toxins of Pseudomonas aeruginosa: new perspectives. Rev. Infect. Dis. 5(Suppl 4): S715-722.
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