تاثیر کوئوروم سنسینگ و تشکیل بیوفیلم بر مقاومت ضد میکروبی و حدت XDR و MDR سودوموناس آئروژینوزا در ماکیان تخم‌گذار

نوع مقاله : مقاله کامل

نویسندگان

چکیده

پیشینه: سودوموناس آئروژینوزا تاثیر عمده‌ای بر شیوع طغیان بیماری‌های طیور دارد. عوامل متعددی ممکن است در بیماری‌زایی آن نقش داشته باشند. هدف: این مطالعه با هدف بررسی شیوع عفونت سودوموناس آئروژینوزا در ماکیان تخم‌گذار و تعیین خصوصیات فنوتیپی و ژنوتیپی جدایه‌ها انجام شد. روش‌ کار: نمونه‌ها (تعداد 160) از ماکیان تخم‌گذار در استان شارکیا، مصر که دچار مشکل تنفسی بودند، با توجه به ضایعه جمع‌آوری شد و برای جداسازی سودوموناس آئروژینوزا اقدام گردید. حساسیت ضد میکروبی بر روی 18 عامل ضد میکروبی انجام شد. ارزیابی کیفی تولید بیوفیلم به روش لوله انجام شد. جدایه‌ها از نظر ژنتیکی برای تشخیص ژن‌های کوئوروم سنسینگ، ژن‌های حدت و ژن‌های تولید بیوفیلم، توسط PCR معمولی مورد بررسی قرار گرفتند. نتایج: سودوموناس آئروژینوزا از 25% نمونه‌ها جدا شد. همچنین 95% جدایه‌ها مقاومت گسترده به دارو (XDR) با شاخص‌های مقاومت آنتی بیوتیکی (MARI) از 67/0 تا 83/0 بودند. در مجموع 38 جدایه توانستند بیوفیلم با درجات مختلف تولید کنند. PCR ژن 16S rRNA و oprL وجود ایزوله سودوموناس آئروژینوزا را تایید کرد. برای ژن‌های کوئوروم سنسینگ، lasI و lasR به ترتیب در 100% و 5/89% با موفقیت تکثیر شدند. برای ژن‌های حدت، toxA و exoU با درصد 9/78% تکثیر شدند، در حالی که ژن higBA در 100% جدایه‌ها حضور داشت. pprA و pprB به ترتیب در 100% و 5/89% نمونه‌ها تکثیر شدند. برای ژن‌های بیوفیلم، pslfliC، و pelA به ترتیب در 100%، 2/84%، و 5/10% نمونه‌ها تکثیر شدند. نتیجه‌گیری: همبستگی قوی بین ژن‌های کوئوروم سنسینگ، ژن‌های بیوفیلم و ژن‌های حدت، قابل تشخیص است. علاوه بر این، تولید بیوفیلم باعث افزایش مقاومت جدایه‌ها در برابر عوامل ضد میکروبی می‌شود.

کلیدواژه‌ها

موضوعات


Abd EL-Dayem, GHA; Ramadan, AH and Ali, HS (2021). The role of hatcheries, hatching eggs and one day old chicks in dissemination of beta lactam antibiotic resistance Pseudomonas aeruginosa (ESBL). Egypt. J. Ani. Health. PP: 80-99.
Abd El-Ghany, WA (2021). Pseudomonas aeruginosa infection of avian origin: Zoonosis and one health implications. Vet. World. 14: 2155-2159.
Abd El-Tawab, AA; El-Hofy, FI; Khater, DF and Al-Adl, MM (2014). PCR detection and gene sequence of Pseudomonas Aeruginosa isolated from broiler chickens. Benha Vet. Med. J., 27: 449-455.
Abdelraheem, WM; Abdelkader, AE; Mohamed, ES and Mohammed, MS (2020). Detection of biofilm formation and assessment of biofilm genes expression in different Pseudomonas aeruginosa clinical isolates. Meta Gene. 23: 100646.
Al-Ahmadi, GJ and Roodsari, RZ (2016). Fast and specific detection of Pseudomonas aeruginosa from other Pseudomonas species by PCR. Ann. Burn. Fire Disasters., 29: 264-267.
Algammal, AM; Eidaroos, NH; Alfifi, KJ; Alatawy, M; Al-Harbi, AI; Alanazi, YF; Ghobashy, MO; Khafagy, AR; Esawy, AM; El-Sadda, SS; Hetta, HF and El-Tarabili, RM (2023). and antibiotic resistancopr L gene sequencing, resistance patterns, virulence genes, quorum snsing e genes of XDR Pseudomonas aeruginosa isolated from broiler chickens. Infect. Drug Resist., 853-867.‏ doi: 10.2147/ IDR.S401473.
Amirmozafari, N; Fallah Mehrabadi, J and Habibi, A (2016). Association of the exotoxin A and exoenzyme S with antimicrobial resistance in Pseudomonas aeruginosa strains. Arch. Iran. Med., 19: 353-358.
Anuj, SN; Whiley, DM; Kidd, TJ; Bell, SC; Wainwright, CE; Nissen, MD and Sloots, TP (2009). Identification of Pseudomonas aeruginosa by a duplex real-time polymerase chain reaction assay targeting the ecfX and the gyrB genes. Diagn. Microbiol. Infect. Dis., 63: 127-131.‏
Badr, JM; El Saidy, FR and Abdelfattah, AA (2020). Emergence of multi-drug resistant Pseudomonas aeruginosa in broiler chicks. Int. J. Microbiol. Biotechnol., 5: 41-47.‏
Bahador, N; Shoja, S; Faridi, F; Dozandeh-Mobarrez, B; Qeshmi, FI; Javadpour, S and Mokhtary, S (2019). Molecular detection of virulence factors and biofilm formation in Pseudomonas aeruginosa obtained from different clinical specimens in Bandar Abbas. Iran. J. Microbiol., 11: 25-30.
Bakheet, AA and Torra, DE (2020). Detection of Pseudomonas aeruginosa in dead chicken embryo with reference to pathological changes and virulence genes. Alex. J. Vet. Sci., 65: 81-89.‏
Bäuerle, T; Fischer, A; Speck, T and Bechinger, C (2018). Self-organization of active particles by quorum sensing rules. Nat. Commun., 9: 1-8.
Bratu, S; Gupta, J and Quale, J (2006). Expression of the las and rhl quorum-sensing systems in clinical isolates of Pseudomonas aeruginosa does not correlate with efflux pump expression or antimicrobial resistance. J. Antimicrob. Chemother., 58: 1250-1253.‏
Chakraborty, S; Dutta, TK; Roychoudhury, P; Samanta, I; Kalai, S and Bandyopadhyay, S (2020). Molecular characterization of biofilm-producing Pseudomonas aeruginosa isolated from healthy pigs and chicken in India. Indian J. Ani. Res., 54: 1400-1407.‏
Christensen, GD; Simpson, WA; Bisno, AL and Beachey, EH (1982). Adherence of slime-producing strains of Staphylococcus epidermidis to smooth surfaces. Infect. Immun., 37: 318-326.‏
Dong, N; Liu, C; Hu, Y; Lu, J; Zeng, Y; Chen, G and Zhang, R (2022). Emergence of an extensive drug resistant Pseudomonas aeruginosa strain of chicken origin carrying bla IMP-45, tet (X6), and tmexCD3-toprJ3 on an IncpRBL16 plasmid. Microbiol. Spect., 10(6): e02283-22.‏
Donnik, IM; Krivonogova, AS; Isaeva, AG; Shkuratova, IA; Moiseeva, KV and Musikhina, NB (2020). Special features of Pseudomonas aeruginosa strains in animal and poultry farms in the regions with various levels of man-made pollution. Agron. Res., 18: 2365-2373.
El-Demerdash, AS and Bakry, NR (2020). Evaluation of the synergistic effect of amikacin with cefotaxime against Pseudomonas aeruginosa and its biofilm genes expression. Gene Exp. Pheno. Traits. 121-138.‏
Elsayed, M; Ammar, A; Al Shehri, ZS; Abd-El Rahman, H and Abd-El Rahman, NA (2016). Virulence repertoire of Pseudomonas aeruginosa from some poultry farms with detection of resistance to various antimicrobials and plant extracts. Mol. Cell. Biol., 62: 124-132.
Eraky, RD; Abd El-Ghany, WA and Soliman, KM (2020). Studies on Pseudomonas aeruginosa infection in hatcheries and chicken. J. Hel. Vet. Med. Soc., 71: 1953-1962.‏
Fadhil, L; Al-Marzoqi, AH; Zahraa, MA and Shalan, AA (2016). Molecular and phenotypic study of virulence genes in a pathogenic strain of Pseudomonas aeruginosa isolated from various clinical origins by PCR: Profiles of genes and toxins. RJPBCS., 7: 590-598.
Finnan, S; Morrissey, JP; O’gara, F and Boyd, EF (2004). Genome diversity of Pseudomonas aeruginosa isolates from cystic fibrosis patients and the hospital environment. J. Clin. Microbiol., 42: 5783-5792.‏
Francis, VI; Stevenson, EC and Porter, SL (2017). Two-component systems required for virulence in Pseudomonas aeruginosa. FEMS Microbiol. Lett., 364: fnx104.‏
Gajdács, M; Baráth, Z; Kárpáti, K; Szabó, D; Usai, D; Zanetti, S and Donadu, MG (2021). No correlation between biofilm formation, virulence factors, and antibiotic resistance in Pseudomonas aeruginosa: results from a laboratory-based in vitro study. Antibiotics. 10: 1134.
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.‏
Gholami, A; Majidpour, A; Talebi-Taher, M; Boustanshenas, M and Adabi, M (2016). PCR-based assay for the rapid and precise distinction of Pseudomonas aeruginosa from other Pseudomonas species recovered from burns patients. JPMH., 57: E81.‏
Haque, S; Ahmad, F; Dar, SA; Jawed, A; Mandal, RK; Wahid, M; Lohani, M; Khan, S; Singh, V and Akhter, N (2018). Developments in strategies for Quorum sensing virulence factor inhibition to combat bacterial drug resistance. Microb. Pathog., 121: 293-302.
Hassan, WH; Ibrahim, AMK; Shany, SAS and Salam, HSH (2020). Virulence and resistance determinants in Pseudomonas aeruginosa isolated from pericarditis in diseased broiler chickens in Egypt. J. Adv. Vet. Ani. Res., 7: 452–463.doi: 10.5455/javar.2020.g441.
Jing, C; Liu, C; Liu, Y; Feng, R; Cao, R; Guan, Z and Yang, G (2021). Antibodies against Pseudomonas aeruginosa alkaline protease directly enhance disruption of neutrophil extracellular traps mediated by this enzyme. Front. Immunol., 12: 649-654.
Karami, P; Khaledi, A; Mashoof, RY; Yaghoobi, MH; Karami, M; Dastan, D and Alikhani, MY (2020). The correlation between biofilm formation capability and antibiotic resistance pattern in Pseudomonas aeruginosa. Gene Reports. 18: 100561.
Khan, MSA; Ahmad, I; Sajid, M and Cameotra, SS (2014). Current and emergent control strategies for medical biofilms. Antibiofilm Agents: From Diagnosis to Treatment and Prevention. 8: 117-159.‏
Khattab, MA; Nour, MS and El-Sheshtawy, NM (2015). Genetic identification of Pseudomonas aeruginosa virulence genes among different isolates. J. Microb. Biochem. Technol., 7: 274-277.
Koneman, EW; Allen, SD; Janda, WM; Schreckenberger, PC and Winn, WCJr (1997). Color atlas and textbook of diagnostic microbiology. 5th Edn., Philadilphia, USA, J. B. Lippincott Co., PP: 1296-1395.
Langendonk, RF; Neill, DR and Fothergill, JL (2021). The building blocks of antimicrobial resistance in Pseudomonas aeruginosa: Implications for current resistance-breaking therapies. Front. Cell. Infect. Microbiol., 11: 665759. doi: 10.3389/fcimb.
Latifi, A; Foglino, M; Tanaka, K; Williams, P and Lazdunski, A (1996). A hierarchical quorum-sensing cascade in Pseudomonas aeruginosa links the transcriptional activators LasR and RhIR (VsmR) to expression of the stationary-phase sigma factor RpoS. Mol. Microbiol., 21: 1137-1146.
Li, M; Guo, N; Song, G; Huang, Y; Wang, L; Zhang, Y and Wang, T (2023). Type II toxin-antitoxin systems in Pseudomonas aeruginosa. Toxins. 15: 164. doi: 10.3390/toxins15020164.
Magiorakos, AP; Srinivasan, A; Carey, RB; Carmeli, Y; Falagas, ME; Giske, CG; Harbarth, 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 pan drug-resistant bacteria: An international expert proposal for interim standard definitions for acquired resistance. Clin. Microbiol. Infect., 18: 268-281.
Matar, GM; Ramlawi, F; Hijazi, N; Khneisser, I and Abdelnoor, AM (2002). Transcription levels of Pseudomonas aeruginosa exotoxin A gene and severity of symptoms in patients with otitis externa. Curr. Microbiol., 45: 350-354.‏
Meliani, A and Bensoltane, A (2015). Review of Pseudomonas attachment and biofilm formation in food industry. Poult. Fish. Wildl. Sci., 3: 2-7.‏
Michalska, M and Wolf, P (2015). Pseudomonas exotoxin A: optimized by evolution for effective killing. Front. Microbiol., 6: 963.
Mohamed, HM; Alnasser, SM; Abd-Elhafeez, HH; Alotaibi, M; Batiha, GES and Younis, W (2022). Detection of β-lactamase resistance and biofilm genes in pseudomonas species isolated from chickens. Microorganisms. 10: 1975. https://doi.org/10.3390/ microorganisms10101975.
Morales, PA; Aguirre, JS; Troncoso, MR and Figueroa, GO (2016). Phenotypic and genotypic characterization of Pseudomonas spp. present in spoiled poultry fillets sold in retail settings. LWT, 73: 609-614.‏
Noomi, BS (2018). Detection of virulence factors of Pseudomonas aeruginosa in different animals by using bacteriological and molecular methods. Iraqi J. Vet. Sci., 32: 205-210.
Pang, Z; Raudonis, R; Glick, BR; Lin, TJ and Cheng, Z (2019). Antibiotic resistance in Pseudomonas aeruginosa: mechanisms and alternative therapeutic strategies. Biotechnol. Adv., 37: 177-192.
Poulsen, BE; Yang, R; Clatworthy, AE; White, T; Osmulski, SJ; Li, L and Hung, DT (2019). Defining the core essential genome of Pseudomonas aeruginosa. PNAS., 116: 10072-10080.‏ doi: 10.1073/pnas.1900570116.
Qin, X; Emerson, J; Stapp, J; Stapp, L; Abe, P and Burns, JL (2003). Use of real-time PCR with multiple targets to identify Pseudomonas aeruginosa and other nonfermenting gram-negative bacilli from patients with cystic fibrosis. J. Clin. Microbiol., 41: 4312-4317.‏
Quinn, PJ; Markey, BK; Carter, ME; Donnelly, WJC and Leonard, FC (2002). Veterinary microbiology and microbial disease. Blackwell Science., 14: 113-118. https://sid.ir/paper/657266/en.
Rodrigue, A; Quentin, Y; Lazdunski, A; Méjean, V and Foglino, M (2000). Cell signalling by oligosaccharides. Two-component systems in Pseudomonas aeruginosa: why so many? Trends Microbiol., 8: 498-504.‏ doi: 10.1016/S0966-842X(00)01833-3.
Roulová, N; Mot’ková, P; Brožková, I and Pejchalová, M (2022). Antibiotic resistance of Pseudomonas aeruginosa isolated from hospital wastewater in the Czech Republic. J. Water Health. 20: 692-701.
Shahat, HS; Mohamed, HM; Abd Al-Azeem, MW and Nasef, SA (2019). Molecular detection of some virulence genes in Pseudomonas aeruginosa isolated from chicken embryos and broilers with regard to disinfectant resistance. Int. J. Vet. Sci., 2: 52-70.
Shukla, S and Mishra, P (2015). Pseudomonas aeruginosa infection in broiler chicks in Jabalpur. Int. J. Ext. Res., 6: 37-39.‏
Skindersoe, ME; Alhede, M; Phipps, R; Yang, L; Jensen, PO; Rasmussen, TB; Bjarnsholt, T; Tolker-Nielsen, T; Høiby, N and Givskov, M (2008). Effects of antibiotics on quorum sensing in Pseudomonas aeruginosa. Antimicrob. Agents Chemother., 52: 3648-3663.
Smith, RS and Iglewski, BH (2003). Pseudomonas aeruginosa quorum sensing as a potential antimicrobial target. JCI., 112: 1460-1465.‏ doi: 10.1172/JCI20364.
Sonbol, F; El-Banna, T; Elgaml, A and Aboelsuod, KM (2022). Impact of quorum sensing system on virulence factors production in Pseudomonas aeruginosa. J. Pure Appl. Microbiol., 16: 1226-1238.‏
Spilker, T; Coenye, T; Vandamme, P and LiPuma, JJ (2004). PCR-based assay for differentiation of Pseudomonas aeruginosa from other Pseudomonas species recovered from cystic fibrosis patients. J. Clin. Microbiol., 42: 2074-2079.‏
Steindler, L; Bertani, I; De Sordi, L; Schwager, S; Eberl, L and Venturi, V (2009). LasI/R and RhlI/R quorum sensing in a strain of Pseudomonas aeruginosa beneficial to plants. Appl. Environ. Microbiol., 75: 5131-5140.‏
Tartor, YH and El-Naenaeey, EY (2016). RT-PCR detection
of exotoxin genes expression in multidrug resistant Pseudomonas aeruginosa. Mol. Cell. Biol., 62: 56-62.‏
Tawakol, M; Nabil, N and Reda, R (2018). Molecular studies on some virulence factors of Pseudomonas aeruginosa isolated from chickens as a biofilm forming bacteria. Journal Article: Assiut Veterinary Medical Journal. 64: 43-51.
Thi, MTT; Wibowo, D and Rehm, BH (2020). Pseudomonas aeruginosa Biofilms. Int. J. Mol. Sci., 21: 8671.
Williams, JJ; Halvorsen, EM; Dwyer, EM; DiFazio, RM and Hergenrother, PJ (2011). Toxin-antitoxin (TA) systems are prevalent and transcribed in clinical isolates of Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus. FEMS Microbiol. Lett., 322: 41-50.‏
Winstanley, C; Kaye, SB; Neal, TJ; Chilton, HJ; Miksch, S; Hart, CA and the Microbiology Ophthalmic Group (2005). Genotypic and phenotypic characteristics of Pseudomonas aeruginosa isolates associated with ulcerative keratitis. J. Med. Microbiol., 54: 519-526.
Wood, TL and Wood, TK (2016). The HigB/HigA toxin/antitoxin system of Pseudomonas aeruginosa influences the virulence factors pyochelin, pyocyanin, and biofilm formation. Microbiol. Open. 5: 499-511.‏ doi: 10.1002/mbo3.346.
Xu, J; Moore, JE; Murphy, PG; Millar, BC and Elborn, JS (2004). Early detection of Pseudomonas aeruginosa-comparison of conventional versus molecular (PCR) detection directly from adult patients with cystic fibrosis (CF). Ann. Clin. Antimicrob., 3: 1-5.‏
Yan, S and Wu, G (2019). Can biofilm be reversed through quorum sensing in Pseudomonas aeruginosa? Front. Microbiol., 10: 1582. doi: 10.3389/fmicb.2019.01582.
Yekani, M; Memar, MY; Alizadeh, N; Safaei, N and Ghotaslou, R (2017). Antibiotic resistance patterns of biofilm-forming Pseudomonas aeruginosa isolates from mechanically ventilated patients. Int. J. Sci. Study. 5: 84-88.
Zhao, X; Yu, Z and Ding, T (2020). Quorum-sensing regulation of antimicrobial resistance in bacteria. Microorganisms. 8: 425.
Zhou, J; Li, S; Li, H; Jin, Y; Bai, F; Cheng, Z and Wu, W (2021). Identification of a toxin-antitoxin system that contributes to persister formation by reducing NAD in pseudomonas aeruginosa. Microorganisms. 9: 753.‏ doi: 10.3390/microorganisms9040753.