CHARACTERIZATION OF CLASS 1 INTEGRONS AND SOME ANTI-MICROBIAL RESISTANCE GENES IN Salmonella SPECIES ISOLAT-ED FROM POULTRY IN EGYPT
Salmonella enterica includes serotypes that were implicated as a food borne pathogens crucially affecting public health and the economic organization. This study was directed to isolate and identify of Salmonella strains from 222 different species and ages of poultry (broiler, chick, ducks, pigeon, quails) from Kafr El Sheikh governorate. The Salmonella isolation rate was (4.5%) as (0.9%) from apparently healthy, (3.6%) from diseased birds. The outer membrane protein F gene was used as promising tool for detection of Genus Salmonella, after that four isolates were identified serologically as two Salmonella enterica serovar Enteritidis and two Salmonella enterica serovar Typhimurium. The resistance pattern of positive Salmonella isolates showed multidrug resistance phenotypes and qnrS for quinolone resistant genes was recorded in one isolate while blaTEM for β-lactam resistant isolates, aacC for aminoglycosides were recorded in all four Salmonella isolates using PCR technique. Also, Class 1 integrons detected with a percentage of (100%) in examined isolates. Sequencing of the class 1 integrons cassettes showed genes encoding resistance specified to streptothricin acetyltransferase (sat) gene, aminoglycoside acetyltransferase (aac3-Id) and aminoglycoside adenyltransferase (aadA7). Class 1 integrons harbored gene encoding domains unfunction protein (duf) in one S. enterica serovar Typhimurium isolate. This study spotlights the significant role of the drug–resistance genes and Class 1 integrons in Salmonella as zoonotically important pathogens of public health importance.
Key words: poultry; Salmonella; drug- resistance genes; integrons gene cassettes
Amagliani G, Brandi G, Schiavano GF. Incidence and role of Salmonella in seafood safety. F Res Int 2012; 45:780â€“8.
Scallan E, Hoekstra RM, Angulo FJ, Tauxe RV, Widdowson MA, Roy SL, Jones JL, Griffin PM. Foodborne illness acquired in the United Statesâ€“major pathogens. Emerg Infect Dis 2011; 17(1):7â€“15.
Antunes P , Mourao J, Campos J, Peixe L. Salmonellosis: The role of poultry meat. Clin Microbiol Infect 2016; 22:110â€“21.
European Food Safety Authority. The Eu-ropean Union summary report on
trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2010. EFSA J 2012; 10(3): 2597.
Baalaji NS, Mathew MK, Krishnaswamy S. Functional assay of Salmonella Typhi ompC using reconstituted large unilamellar vesicles: a general method for characterization of outer membrane proteins. Bio chemie 2006; 88: 1419â€“24.
Williams KM, Bigley EC, Raybourne RB. Identification of murine B-cell and T-cell epitopes of Escherichia coli outer membrane protein F with synthetic polypeptides. Infect Immun 2000; 68:2535â€“45.
Abd El- Tawab AA, Ahmed MA, Amany IE, Mai ME. A study of outer membrane protein (OMPs) genes for detection of salmonella or-ganisms in poultry farms. BVMJ 2016; 30 (1): 231â€“7.
Ben Salem R, Abbassi MS, Garcia V, Garcia-Fierro R, Fernandez J, Kilani
H, Jaouani I, Khayeche M, Messadi L, Rodi-cio MR. Antimicrobial drug resistance and ge-netic properties of Salmonella enterica serotype Enteritidis circulating in chicken farms in Tuni-sia. J Infect Public Health 2017; 10 (6): 855â€“60.
Taddele MH, Rathore R, Dhama K. Anti-biogram assay of S.Gallinarum and other S. Enteric serovars of poultry origin in India. J Anim Vet Adv 2012; 7: 309â€“17.
Kim JH, Cho JK, Kim KS. Prevalence and characterization of plasmid-mediated quin-olone resistance genes in Salmonella isolated from poultry in Korea. Avian Pathol 2013; 42(3): 221â€“9.
Frye JG, Jackson CR . Genetic mecha-nisms of antimicrobial resistance identified in Salmonella enterica, Escherichia coli, and En-teroccocus spp. isolated from U.S. food ani-mals. Front Microbiol 2013; 4: 135.
Trongjit S, Angkititrakul S, Tuttle RE, Poungseree J, Padungtod P, Chuanchuen R. Prevalence and antimicrobial resistance in Sal-monella enterica isolated from broiler chickens, pigs and meat products in Thailand-Cambodia border provinces. Microbiol Immunol 2017; 61(1): 23â€“33.
Kakatkar AS, Pansare LS, GautamRK, Shashidhar R, Karani M, Bandekar JR. Molecu-lar characterization of antibiotic resistant Sal-monella isolates from Indian foods. Food Res Int 2011; 44(10):3272â€“5.
Phongpaichit S, Wuttananupan K, Sama-santi W. Class1 integrons and multidrug re-sistance among Escherichia coli isolates from human stools. Southeast Asian J Trop Med Pub-lic Health 2008; 39 (2): 279â€“87.
Carattoli A. Importance of integrons in the diffusion of resistance. Veterinary Re-search, BioMed Central 2001; 32 (3-4): 243â€“59.
Labbate M, Case RJ, Stokes HW. The in-tegron gene cassette system: an active player in bacterial adaptation. Methods Mol Biol 2009; 532: 103â€“25.
Leverstein-van MA, Blok HE, Donders AR, Paauw A, Fluit AC, Verhoef J.
Multidrug resistance among Enterobacteri-aceae is strongly associated with the presence of integrons and is independent of species or isolate origin. J Infect Dis 2003; 187 (2): 251â€“9.
International Standards Organization (ISO 6579). General guidance on methods for the detection of Salmonella, Geneva, Switzer-land, 2002.
Kauffman G. Kauffmann white scheme. J Acta Path Microbiol Sci 1974; 61: 38.
Tatavarthy A, Cannons A. Real-time PCR detection of Salmonella species using a novel target: the outer membrane porin F gene (ompF). Lett Appl Microbiol 2010; 50 (6): 645â€“52.
Finegold SM, Martin WJ. Diagnostic mi-crobiology. 6th Ed., The C.V. Mosby Company, St. Louis, Toronto, London, 1982.
CLSI. Performance Standards for Anti-microbial Susceptibility Testing; Twenty- Fifth Informational Supplement (M100 - S25), Clini-cal and Laboratory Standards Institute. Pennsyl-vania 19087, USA, 2015; 44â€“50Ø²
Lynne AM, Rhodes-Clark BS, Bliven K, Zhao S, Foley SL. Antimicrobial Resistance Genes Associated with Salmonella enterica serovar Newport isolates from Food Animals. Antimicrob Agents Chemother 2008; 52(1): 353â€“6.
Robicsek A, Strahilevitz J, Jacoby GA, Macielag M, Abbanat D, Park CH, Bush K, Hooper DC. Fluoroquinolone modifying en-zyme: a new adaptation of a common amino-glycoside acetyltransferase. Nat MED 2006; 12 (1): 83 â€“88.
Colom K, PÃ¨rez J, Alonso R, FernÃ¡ndez-Aranguiz A, LariÅˆo E, Cisterna R. Simple and reliable multiplex PCR assay for detection of blaTEM, blaSHV and blaOXA-1 genes in En-terobacteriaceae. FEMS Microbiol Lett 2003; 223 (2):147â€“51.
Sow AG, Wane A, Diallo MH, Boye CS, AÃ¯dara-Kane A. Genotypic characterization of antibiotic-resistant Salmonella Enteritidis iso-lates in Dakar, Senegal. J Infect Developing Countries 2007; 1 (3): 284â€“8.
Hall A. BioEdit: a user-friendly biologi-cal sequence alignment editor and analysis pro-gram for Windows 95/98/NT. Nuc Acids Symp Ser 1999; 41: 95â€“8.
Barrow PA, Freitas Neto OC. Pullorum disease and fowl typhoid new thoughts on old diseases. Avian Pathol 2011; 40 (1): 1-13.
Clothier KA, Kim P, Mete A, Hill AE. Frequency, serotype distribution and antimicro-bial susceptibility patterns of Salmonella in small poultry flocks in California. J Vet Diagn Invest 2018; 30 (3): 471- 475.
Akbarmehr J. Isolation of Salmonella spp. from poultry (ostrich, pigeon, and
chicken) and detection of their hilA gene by PCR method. A J Microbiol Res 2010; 4 (24): 2678-2681.
Lebdah MA, Waffa MM, Samah E, Re-hab IH. Molecular detection of some
Antimicrobial Resistance Genes in Salmonel-la species isolated from commercial layers in Egypt. Zagazig Vet J 2017; 45(1): 29-38.
Liu W B, Chen J, Huang YY, Liu B, ShiX M. Serotype, genotypeand antimicrobial susceptibility profiles of Salmonella from chicken farms in Shanghai. J Food Prot 2010; 73(3): 562-567.
Abd El-Ghany WA, Soumaya SA, El-ShafiiHatem ME. A survey on Salmonella spe-cies isolated from chicken flocks in Egypt. AJAVA 2012; 7 (6): 489-501.
Ibrahim MA, Emeash HH, Nahed HG, Abdel-Halim MA. Seroepidemiological Studies on Poultry Salmonellosis and its Public Health Importance. J World's Poult Res 2013; 3 (1): 18-23.
AL-Iedani AA, Khudor MH, Oufi NM. Isolation and identification of Salmonella spp. from poultry farms by using different tech-niques and evaluation of their
antimicrobial susceptibilities. Bas J Vet Res 2014; 13(1): 246-259.
Dhaher FH, Awni MDH, Mahmood NR, Jamil MM. Isolation and diagnosis of
Salmonella in Animal origin food. Iraq Acad Sci J 2011; 3(5):1-19.
Marwa RA. Molecular characterization of some Antibiotic Resistance Genes in Salmo-nella Species isolated from diseased poultry. MVSc thesis, Fac of Vet Med, kafrelsheikh Univ, Egypt, 2016.
Orady M R, Salwa MH, Ahmed MAA, Wafaa MH, Etab MA, Azza S E. Molecular Characterization of Class 1 Integrons and Anti-biotic Resistance Genes in Salmonella enterica isolated from Chicken. Global Veterinaria 2017; 18 (5): 322â€“31.
Gast R K. Paratyphoid infections. In D. E. Swayne (Ed.), Diseases of Poultry,
Ames, IA: Wiley-Blackwell Publishing 2013; 693â€“733.
Neubauer H, Hensel A, Aleksic S, Meyer H. Identification of Yersinia enterocolitica with-in the genus Yersinia. Syst Appl Microbiol 2000; 23(1): 58â€“62.
Mohamed NS. A survey of Salmonella contamination in chicken carcass and giblets in central Ethiopia. Revue de Med Vet 2003; 154(4):267â€“70.
Ahmed AM, Shimamoto T. Genetic analysis of multiple antimicrobial resistance in Salmonella isolated from diseased broilers in Egypt. Microbiol Immunol 2012; 56 (4): 254â€“61.
GarcÃa C, Soriano JM, BenÃtez V, CatalÃ¡-Gregori P. Assessment of Salmonella spp. in feces, cloacal swabs, and eggs (eggshell and content separately) from a laying hen farm. Poult Sci 2011; 90 (7):1581â€“5.
Ren D, Chen P, Wang Y, Wang J, Liu H, Liu H. Phenotypes and antimicrobial
resistance genes in Salmonella isolated from retail chicken and pork in Changchun, China. J Food Saf 2017; 37 (2): e12314.
Zhu Y, Lai H, Zou L, Yin S, Wang C, Han X, Xia X, Hu K, He L, Zhou K, Chen S, Ao X, Liu S. Antimicrobial resistance and re-sistance genes in Salmonella strains isolated from broiler chickens along the slaughtering process in China. Int J Food Microbiol 2017; 259:43â€“51.
Begum K, Mannan SJ, Ahmed A. Anti-biotic Resistance, Plasmids and Integron Profile of Salmonella Species Isolated from Poultry Farm and Patients. Dhaka Univ. J Pharm Sci 2016; 15(2): 209â€“14.
Asif M, Rahman H, Qasim M, Khan TA, Ullah W, Jie Y. Molecular detection and antimi-crobial resistance profile of zoonotic Salmonel-la Enteritidis isolated from broiler chickens in Kohat, Pakistan. J Chin Med Assoc 2017; 80(5):303â€“6.
Abdel-Maksoud M, Abdel-Khalek R, El-Gendy A, Gamal RF, Abdelhady HM, House BL. Genetic characterisation of multidrug-resistant Salmonella enterica serotypes isolated from poultry in Cairo, Egypt. Afr J Lab Med 2015; 4 (1):158â€“65.
Abatcha MG, Mohd EE, GulamR. Preva-lence, antimicrobial resistance, resistance genes and class 1 integrons of Salmonella serovars in leafy vegetables, chicken carcasses and related processing environments in Malaysian fresh food markets. Food Control 2018; 91: 170â€“80.
Ãlvarez- Fernandez E, Alonso-Calleja C,GarcÃa-FernÃ¡ndez C, Capita R. Prevalence and antimicrobial resistance of Salmonella sero-types isolated from poultry in Spain: compari-son between
and 2006. Int J food Microbiol 2012; 153 (3): 281â€“7.
Ahmed AM, Younis EE, Ishida Y, Shimamoto T. Genetic basis of multidrug re-sistance in Salmonella enterica serovars Enter-itidis and Typhimurium isolated from diarrheic calves in Egypt. Acta Trop 2009; 111(2): 144â€“9.
Elmonir W, Hegazym AM, El-Tras WF, Shohiep A. Extremely drug-resistant Salmonella in broiler production chain in Egypt. Life Sci J 2017; 14(9): 82â€“7.
Zishiri OT, Mkhize N, Mukaratirwa S. Prevalence of virulence and antimicrobial re-sistance genes in Salmonella spp. isolated from commercial chickens and human clinical iso-lates from South Africa and Brazil. Onder-stepoort J Vet Res 2016; 83(1): 1â€“11.
Gharieb RM, Tartor YH, Khedr MH. Non-Typhoidal Salmonella in poultry meat and diarrhoeic patients: prevalence, antibiogram, virulotyping, molecular detection and sequenc-ing of class I integrons in multidrug resistant strains. Gut Pathog 2015;7:34.
Lee MF, Chen YH, Peng CF. Molecular characterisation of class 1 integrons in Salmo-nella enterica serovar Choleraesuis isolates from southern Taiwan. Int J Antimicrob Agents 2009; 33(3): 216â€“22.
De Lappe N, O'Halloran F, Fanning S, Corbett-Feeney G, Cheasty T, Cormican M. Antimicrobial resistance and genetic diversity of Shigella sonnei isolates from western Ireland, an area of low incidence of infection. J Clin Microbiol 2003; 41(5):1919â€“24.
Alghoribi MF, Balkhy H, Doumith M, Al Johani SM, Upton M, Woodford N, Ellington MJ. Molecular Epidemiology, Virulence Poten-tial and Antibiotic Susceptibility of the Major Lineages of Uropathogenic Escherichia coli. Ph.D. thesis, Faculty of Medical and Human Sciences, Manchester Univ, 2015.
Ye L, Li R, Lin D, Zhou Y, Fu A, Ding Q, Chan EW, Yao W, Chen S. Characterization of an IncA/C Multidrug Resistance Plasmid in Vibrio alginolyticus. Antimicrob Agents Chemother 2016; 60(5):3232â€“5.
Goodacre NF, Gerloff DL, Uetz P. Pro-tein domains of unknown function are essential in bacteria. mBio 2013; 5(1):e00744â€“13.