Correlation between ESβL Salmonella Serovars Isolated from Broilers and their Virulence Genes

Salmonella virulence ESBL Broilers genes

Salmonella is considered to be one of the major poultry bacterial pathogens. The poultry species are one of the main reservoirs for the human types, thus serving as public health hazards. The development of drug resistant genes and multidrug resistant types from Salmonella has increased recently. This current study was undertaken to estimate the correlation between extended spectrum beta-lactamase multidrug resistant (ESβL) Salmonella serovars isolated from broilers and their virulence genes. Two hundred and forty samples were collected from clinically diseased broilers chicks (showed disorders of the intestinal tract) and examined for the presence of Salmonella isolates according to ISO 6579: 2002 and ISO. 6579-3:2014 Fifty Salmonella isolates were isolated with an incidence of 20.8%. Isolates of Salmonella were serotyped as follows: 25 S. Kentucky, 9 S. Infantis, 6 S. Enteritidis, 4 S. Heidelberg, and one isolates per serovars S. Labadie, S. Typhi, S. Agona, S. Pullorum, S. Newport and S. Virginia. AST (antimicrobial susceptibility testing) showed that high percentage of isolates were resistant to all Ampicillin (90%), Nalidixic acid (88%), Sulfamethoxazole + Trimethoprim (82%) and Tetracycline (82%). Approximately 86% of the isolates demonstrated multiple resistance, of which 18.75% and 25% were resistant to three and four antimicrobial types, respectively. Phenotypic detection of ESβLs by using screening test (Cefinase®) and confirmatory test by using combined disk diffusion test revealed that 32% of isolates were positive for both tests with 20% similarity and 12% diversity between the two tests. Molecular characterization of some ESβLs genes (blaTEM, blaCTX, blaOXA, blaCMY and blaSHV) and some virulence genes (invA, avrA, sopB, bcfC, stn (was done using PCR. The results showed that all the ESβLs positive serovars were positive for amplification of all tested virulence genes and noticed that all the isolates were negativefor blaCMY gene. The present study suggests that virulent ESβL Salmonella serovars could infect broilers and should be taken into consideration as an important bacterial pathogen affecting poultry.

Article Details
  • Section
  • Research Articles
Download data is not yet available.
Abouelmaatti, R. R.; Algammal, A. M.; Li, X.; Ma, J.; Abdelnaby, E. A. and Elfeil, W. M. K. (2013):Cloning and analysis of Nile tilapia Tolllike receptors type-3 mRNA Central European Journal of Immunology. 3: 277-282.
Amini, K.; Salehi, T. Z.; Nikbakht, G.; Ranjbar, R.; Amini, J. and Ashrafganjooei, S. B. (2010):Molecular detection of invA and spv virulencegenes in Salmonella enteritidis isolated from human and animals inIran African Journal of Microbiology Research. 4: 2202-2210.
Authority, E. F. S. (2009):Analysis of the baseline survey on the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in holdings with breeding pigs, in the EU, 2008. Part A: MRSA prevalence estimates; on request from the European Commission EFSA J. 7: 1376.
Ayoub, M. A.; Elfeil, W. K.; El Boraey, D.; Hammam, H. and Nossair, M. A. (2019):Evaluation of Some Vaccination Programs in Protection of Experimentally Challenged Broiler Chicken against Newcastle Disease Virus American Journal of Animal and Veterinary Sciences. 14:197-206.
Ben-Barak, Z.; Streckel, W.; Yaron, S.; Cohen, S.; Prager, R. and Tschäpe, H. (2006):The expression of the virulence-associated effector protein gene avrA is dependent on a Salmonella enterica-specific regulatory function International journal of medical microbiology. 296: 25-38.
Brinas, L.; Moreno, M. A.; Zarazaga, M.; Porrero, C.; Sáenz, Y.; García, M.; Dominguez, L. and Torres, C. (2003):Detection of CMY-2, CTX-M-14, and SHV-12 β-lactamases in Escherichia coli fecal-sample isolates from healthy chickens Antimicrobial agents and chemotherapy. 47: 2056-2058.
Campioni, F.; Bergamini, A. M. M. and Falcão, J. P. (2012):Genetic diversity, virulence genes and antimicrobial resistance of Salmonella Enteritidis isolated from food and humans over a 24-year period in Brazil Food microbiology. 32: 254-264.
Carattoli, A. (2003):Plasmid-mediated antimicrobial resistance in Salmonella enterica Current issues in molecular biology. 5: 113-122.
Cesco, M. D. O.; Zimermann, F.; Giotto, D.; Guayba, J.; Borsoi, A.; Rocha,
S.; Moraes, H. and Nascimento, V. (2009):Survey of virulence genes in Salmonella Hadar in materials coming from chickens Veterinaria em foco. 6: 159-164.
Chen, S.; Zhao, S.; White, D. G.; Schroeder, C. M.; Lu, R.; Yang, H.; Mcdermott, P. F.; Ayers, S. and Meng, J. (2004):Characterization of multiple-antimicrobial-resistant salmonella serovars isolated from retail meats Appl Environ Microbiol. 70: 1-7.
Crăciunaş, C.; Keul, A.-L.; Flonta, M. and Cristea, M. (2012):DNA-based diagnostic tests for Salmonella strains targeting hilA, agfA, spvC and sef genes Journal of environmental management. 95: S15-S18.
Diab, M. S.; Abd El Hafez, M. S.; Ashry, M. A. and Elfeil, W. K. (2019):Occurrence of Avian Influenza H5N1 among Chicken, Duck Farms and Human in Egypt American Journal of Animal and Veterinary Sciences. 14: 26-32.
Dione, M. M.; Ikumapayi, U.; Saha, D.; Mohammed, N. I.; Adegbola, R. A.; Geerts, S.; Ieven, M. and Antonio, M. (2011):Antimicrobial resistance and virulence genes of non-typhoidal** Salmonella** isolates in The Gambia and Senegal Journal of infection in developing countries. 5: 765-775.
Efsa. (2018):The European Union summary report on antimicrobial esistance in zoonotic and indicator bacteria from humans, animals and food in 2016, European Food Safety Authority and European Centre for Disease Prevention Control. p. e05182.
Efsa., E. (2008):Report of the task force on zoonoses data collection on the analysis of the baseline survey on the prevalence of Salmonella in slaughter pigs, in the EU, 2006-2007, part A: Salmonella prevalence estimates The EFSA Journal. 135: 1-111.
Eid, H. M.; Algammal, A. M.; Elfeil, W. K.; Youssef, F. M.; Harb, S. M. and Abd-Allah, E. M. (2019):Prevalence, molecular typing, and antimicrobial resistance of bacterial pathogens isolated from ducks Veterinary World. 12: 677-683.
Elfeil, W. K.; Abouelmaatti, R. R.; Sun, C. J.; Han, W. Y.; Li, X. K.; Ma, J. S.; Lei, L. C.; Liu, S. S.; Yang, Y. J.; Wang, Y.; Mandour, M.; Fawzy, M. and Shalaby, M. N. (2012):Identification, Cloning, Expression of a Novel Functional Anas platyrhynchos mRNA TLR4 Journal of Animal and Veterinary Advances. 11: 1727-1733.
Elfeil, W. K.; Ezzat, M. E.; Fathi, A.; Alkilany, M.-a. A. and Abouelmaatti, R. R. (2020):Prevalence and Genotypic Analysis and Antibiotic Resistance of Salmonella Species Isolated from Imported and Freshly Slaughtered Chicken American Journal of Animal and Veterinary Sciences. 15: 134-144.
Elfeil, W. M.; Algammal, A. M.; Abouelmaatti, R. R.; Gerdouh, A. and Abdeldaim, M. (2016):Molecular characterization and analysis of TLR-1 in rabbit tissues Cent Eur J Immunol. 41: 236-242.
Elhady, M. A.; Ali, A.; Kilany, W. H.; Elfeil, W. K.; Ibrahim, H.; Nabil, A.; Samir, A. and El Sayed, M. (2018):Field Efficacy of an Attenuated Infectious Bronchitis Variant 2 Virus Vaccine in Commercial Broiler Chickens Vet Sci. 5.
Enany, M. E.; Algammal, A. M.; Shagar, G. I.; Hanora, A. M.; Elfeil, W. K. and Elshaffy, N. M. (2018):Molecular typing and evaluation of Sidr honey inhibitory effect on virulence genes of MRSA strains isolated from catfish in Egypt Pak J Pharm Sci. 31: 1865-1870.
Fawzy, M.; Ali, R.; Elfeil, W.; Saleh, A.; Mady, W. and Eltarabilli, M. (2020):Efficacy of inactivated velogenic Newcastle disease virus genotype VII vaccine in broiler chickens Veterinary Research Forum.
Gast, R. K.; Guard-Petter, J. and Holt, P. S. (2003):Effect of prior serial in vivo passage on the frequency of Salmonella enteritidis contamination in eggs from experimentally infected laying hens Avian diseases. 47: 633-639.
Grimont, P. and Weill, F.-X. (2007): Antigenic Formulae of the Salmonella
serovars, (9th ed.) Paris: WHO Collaborating Centre for Reference and Research on Salmonella Institute Pasteur. 1-166.
Groisman, E. A. and Ochman, H. (1996):Pathogenicity islands: bacterial evolution in quantum leaps Cell. 87: 791-794.
Hacker, J.; Blum‐Oehler, G.; Mühldorfer, I. and Tschäpe, H. (1997):Pathogenicity islands of virulent bacteria: structure, function and impact on microbial evolution Molecular microbiology. 23: 1089-1097.
Hegazy, W. a. H. and Hensel, M. (2012):Salmonella enterica as a vaccine carrier Future microbiology. 7: 111-127.
Helms, M.; Simonsen, J. and Mølbak, K. (2004):Quinolone resistance is associated with increased risk of invasive illness or death during infection with Salmonella serotype Typhimurium Journal of Infectious Diseases. 190: 1652-1654.
Helms, M.; Vastrup, P.; Gerner-Smidt, P. and Mølbak, K. (2002): Excess mortality associated with antimicrobial drug-resistant Salmonella Typhimurium Emerging infectious diseases. 8:490.
Hopkins, K. L. and Threlfall, E. J. (2004):Frequency and polymorphism of sopE in isolates of Salmonella enterica belonging to the ten most prevalent serotypes in England and Wales Journal of medical microbiology. 53: 539-543.
Hughes, L. A.; Shopland, S.; Wigley, P.; Bradon, H.; Leatherbarrow, A. H.; Williams, N. J.; Bennett, M.; De Pinna, E.; Lawson, B. and Cunningham, A. A. (2008):Characterisation of Salmonella enterica serotype Typhimurium isolates from wild birds in northern England from 2005–2006 BMC Veterinary Research. 4:4.
Lee, L. A.; Puhr, N. D.; Maloney, E. K.; Bean, N. H. and Tauxe, R. V. (1994):Increase in antimicrobial-resistant Salmonella infections in the United States, 1989–1990 Journal of Infectious Diseases. 170:128-134.
Liu, Y.; Yang, Y.; Liao, X.; Li, L.; Lei, C.; Li, L.; Sun, J. and Liu, B. (2012):Antimicrobial resistance, resistance genes and virulence genes in Salmonella isolates from chicken J. Anim. Vet. Adv. 11:4423-4427.
M. Algamma, A.; I. Eid, H.; A. Nasef, S.; Elfeil, W. K. and H. Mansour, G. (2016):Genetic Variation among Avian Pathogenic E. coli Strains Isolated from Broiler Chickens Asian Journal of Animal and Veterinary Advances. 11: 350-356.
Mohammed, L.; Samaha, H.; Draz, A. and Haggag, Y. (1999):Salmonellae among birds and human beings Alex. J. Vet. Sc. 15: 147-154.
Nccls. (2015):performance standards for Antimicrobial Susceptibility Testing;fifteenth Informational Supplement According to CLSI. CLSI document M100-s15., Clinical Laboratory standard Institue, Wayne.
Prager, R.; Mirold, S.; Tietze, E.; Strutz, U.; Knüppel, B.; Rabsch, W.; Hardt, W.-D. and Tschäpe, H. (2000):Prevalence and polymorphism of genes encoding translocated effector proteins among clinical isolates of Salmonella enterica International journal of medical microbiology. 290: 605-617.
Rahman, H.; Streckel, W.; Prager, R. and Tschape, H. (2004):Presence of sopE gene & its phenotypic expression among different serovars of Salmonella isolated from man & animals Indian Journal of Medical Research. 120:35.
Sedeik, M. E.; Awad, A. M.; Rashed, H. and Elfeil, W. K. (2018): Variations in Pathogenicity and Molecular Characterization of Infectious Bursal Disease Virus (IBDV) In Egypt American Journal of Animal and Veterinary Sciences. 13: 76-86.
Sinha, P.; Sharma, R.; Rishi, S.; Sharma, R.; Sood, S. and Pathak, D. (2008):Prevalence of extended spectrum beta lactamase and AmpC beta lactamase producers among Escherichia coli isolates in a tertiary care hospital in Jaipur Indian Journal of Pathology and Microbiology. 51:367.
Standardization, I. O. F. (2007): Microbiology of food and animal feeding stuffs–Horizontal method for the detection of Salmonella spp.–Amendment 1: Annex D: Detection of Salmonella spp. in animal faeces and in environmental samples from the primary production stage. ISO 6579: 2002/Amd. 1: 2007, International Organization for Standardization Geneva, Switzerland.
Standardization, I. O. F. (2014):ISO/TR 6579-3: 2014. Microbiology of the food chain. Horizontal method for the detection, enumeration and serotyping of Salmonella. Part 3: guidelines for serotyping of Salmonella spp, International Organization for Standardization Geneva, Switzerland.
Streckel, W.; Wolff, A. C.; Prager, R.; Tietze, E. and Tschäpe, H. (2004):Expression profiles of effector proteins SopB, SopD1, SopE1, and AvrA differ with systemic, enteric, and epidemic strains of Salmonella enterica Molecular nutrition & food research. 48: 496-503.
Sultan, H. A.; Ali, A.; El Feil, W. K.; Bazid, A. H. I.; Zain El-Abideen, M. A. and Kilany, W. H. (2019a):Protective Efficacy of Different Live Attenuated Infectious Bronchitis Virus Vaccination Regimes Against Challenge With IBV Variant-2 Circulating in the Middle East Frontiers in Veterinary Science. 6:341.
Sultan, H. A.; Arafa, A.-E.; Talaat, S.; Gaballa, A. A.; Kilany, W. H.; Elfeil, W. K.; Shehata, A. A. and Amarin, N. (2019b):Efficacy of Clade 2.3.2 H5-Recombinant Baculovirus Vaccine in Protecting Muscovy and Pekin Ducks from Clade H5N8 Highly Pathogenic Avian Influenza Infection Avian Diseases. 63:219-229, 11.
Sultan, H. A.; Talaat, S.; Elfeil, W. K.; Selim, K.; Kutkat, M. A.; Amer, S. A. and Choi, K.-S. (2020):Protective efficacy of the Newcastle disease virus genotype VII–matched vaccine in commercial layers Poultry Science. 99: 1275-1286.
White, D. G.; Zhao, S.; Sudler, R.; Ayers, S.; Friedman, S.; Chen, S.; Mcdermott, P. F.; Mcdermott, S.; Wagner, D. D. and Meng, J. (2001):The isolation of antibiotic-resistant Salmonella from retail ground meats New England journal of medicine. 345:1147-1154.
Winokur, P.; Brueggemann, A.; Desalvo, D.; Hoffmann, L.; Apley, M.; Uhlenhopp, E.; Pfaller, M. and Doern, G. (2000):Animal and human multidrug-resistant, cephalosporin-resistant Salmonella isolates expressing a plasmid-mediated CMY-2 AmpC β-lactamase Antimicrobial agents and chemotherapy. 44:2777-2783.
Wood, M. W.; Jones, M. A.; Watson, P. R.; Hedges, S.; Wallis, T. S. and Galyov, E. E. (1998):Identification of a pathogenicity island required for Salmonella enteropathogenicity Molecular microbiology. 29:883-891.
Zou, W.; Al-Khaldi, S. F.; Branham, W. S.; Han, T.; Fuscoe, J. C.; Han, J.; Foley, S. L.; Xu, J.; Fang, H. and Cerniglia, C. E. (2010): Microarray analysis of virulence gene profiles in Salmonella serovars from food/food animal environment The Journal of Infection in Developing Countries. 5: 94-105.