RAPD Fingerprinting and Genetic Diversity of Salmonella Spp. Isolated from Broiler and Layer Flocks in Karbala, Iraq

Document Type : Original Articles

Authors

1 Ministry of Iraqi Agriculture, Karbala, Iraq

2 Department of Microbiology, College of Veterinary Medicine, University of Baghdad, Baghdad, Iraq

Abstract

Salmonellosis in poultry is one of the most significant bacterial infections causing mortality, reduced production, and serious economic losses. This study aimed to study the molecular diversity among Salmonella isolates and investigate the epidemiological spread of these bacteria in broiler and layer chicken flocks in five different farms in Karbala, Iraq, using random amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR). In total, 217 cloac a swabs were collected from the farms, out of which 129 and 88 swabs were taken from broiler and layer chickens. The samples were screened by PCR for S. enterica subsp. enterica using primers specific for the invA gene. Afterward, RAPD-PCR with uniplex or multiplex octamer primers was applied to genotype the isolates. The incidence rate of Salmonella infections in broilers and layers was estimated to be 27.9% and 12.5%. The uniplex primers P2 and P3, along with the multiplex primers yielded discriminatory patterns. Moreover, the RAPD typing showed a diverse range of banding patterns of Salmonella spp. Dendrograms created through GelJ software revealed various Salmonella genotypes in broilers and layers. The RAPD-PCR could be used as an accurate and fast tool to identify genetic relatedness among Salmonella spp. The obtained results would assist researchers in epidemiological studies and controlling salmonellosis in poultry fields.

Keywords

Main Subjects

References

  1. Haider M, Hossain M, Hossain M, Chowdhury E, Das P, Hossain M. Isolation and characterization of enterobacteria associated with health and disease in Sonali chickens. Bangladesh J Vet Med. 2004;2(1):15-21.
  2. Yoshida C, Gurnik S, Ahmad A, Blimkie T, Murphy SA, Kropinski AM, et al. Evaluation of molecular methods for identification of Salmonella serovars. J Clin Microbiol. 2016;54(8):1992-8.
  3. Rodriguez J, Rondón I, Verjan N. Serotypes of Salmonella in Broiler Carcasses Marketed at Ibague, Colombia. Braz J Poult Sci. 2015;17:545-52.
  4. Ibrahim WA, Abd El-Ghany WA, Nasef SA, Hatem M. A comparative study on the use of real time polymerase chain reaction (RT-PCR) and standard isolation techniques for the detection of Salmonellae in broiler chicks. Int J Vet Sci. 2014;2(1):67-71.
  5. Saif Y, Fadly A, Glisson J, McDougald L, Nolan L, Swayne D. Diseases of poultry. Ames. Iowa, USA: Blackwell Publisng; 2008.
  6. Control CfD, Prevention. Reports of Salmonella outbreak investigations from 2018. 2018.
  7. Taddele MH, Rathore R, Dhama K, Agarwal R. Epidemiological characterization of Salmonella gallinarum isolates of poultry origin in India, employing two PCR based typing methods of RAPD-PCR and PCR-RFLP. Asian J Anim Vet Adv. 2011;6(11):1037-51.
  8. Altwegg M, Hickman-Brenner F, Farmer III J. Ribosomal RNA gene restriction patterns provide increased sensitivity for typing Salmonella typhi strains. J Infect Dis. 1989;160(1):145-9.
  9. Navarro F, Llovet T, Echeita M, Coll P, Aladuena A, Usera M, et al. Molecular typing of Salmonella enterica serovar Typhi. J Clin Microbiol. 1996;34(11):2831-4.
  10. Thong K-L, Puthucheary S, Yassin RM, Sudarmono P, Padmidewi M, Soewandojo E, et al. Analysis of Salmonella typhi isolates from Southeast Asia by pulsed-field gel electrophoresis. J Clin Microbiol. 1995;33(7):1938-41.
  11. Thangaraj M, Vishruth P, Ramesh T, Lipton A. RAPD fingerprinting and demonstration of genetic variation in three pathogens isolated from Mangrove environment. Asian J Biotechnol. 2011;3(3):269-74.
  12. Cheah Y-K, Salleh NA, Lee L-H, Radu S, Sukardi S, Sim J-H. Comparison of PCR fingerprinting techniques for the discrimination of Salmonella enterica subsp. enterica serovar Weltevreden isolated from indigenous vegetables in Malaysia. World J Microbiol Biotechnol. 2008;24(3):327-35.
  1. Gomes AR, Muniyappa L, Krishnappa G, Suryanarayana V, Isloor S, Prakash B, et al. Genotypic characterization of avian Escherichia coli by random amplification of polymorphic DNA. Int J Poult Sci. 2005.
  2. Hassanien H. Use of randomly amplified polymorphic DNA (RAPD) analysis to detect genetic variation in Seabass Dicentrarchus labrax. J Fish Aquat Sci. 2008;3:39-46.
  3. Maripandi A, Suresh S, Ponmurugan P, Gurusubramanian G. Random amplification of polymorphic DNA (RAPD) of Salmonella enteritidis isolated from chicken samples. Biotechnology. 2007;6(2):278-82.
  4. Idil N, Bilkay IS. Application of RAPD-PCR for determining the clonality of methicillin resistant Staphylococcus aureus isolated from different hospitals. Braz Arch Biol Technol. 2014;57:548-53.
  5. R Jaffer M, K Nazal K. Contamination of local laying hen’s egg shell with salmonella serotypes. Iraq J Vet Med. 2013;37(1):13-6.
  6. Heras J, Domínguez C, Mata E, Pascual V, Lozano C, Torres C, et al. GelJ–a tool for analyzing DNA fingerprint gel images. BMC Bioinform. 2015;16(1):1-8.
  7. Al-Abadi I, Al-Mayah A. Isolation and identification of Salmonella spp. from chicken and chicken environment in Basrah province. Afr J Biol Sci. 2011;7:33-43.
  8. Meteab BK, Abed AAA. Isolation and identification of Salmonella serotypes in poultry. Al-Qadisiyah J Vet Med Sci. 2018;17(1):75-80.
  9. Taha ZM, Ahmed MS, Abdo JM. Occurrence and antimicrobial resistance of Salmonella serotypes isolated from chicken carcasses in Duhok, Kurdistan Region/Iraq. 2015;17: 119-128.
  10. Betancor L, Schelotto F, Martinez A, Pereira M, Algorta G, Rodríguez MA, et al. Random amplified polymorphic DNA and phenotyping analysis of Salmonella enterica serovar Enteritidis isolates collected from humans and poultry in Uruguay from 1995 to 2002. J Clin Microbiol. 2004;42(3):1155-62.

23.          Quintaes BR, Leal NC, Reis EMF, Hofer E. Optimization of randomly amplified polymorphic DNA-polymerase chain reaction for molecular typing of Salmonella enterica serovar Typhi. Rev Soc Bras Med. 2004;37:143-7. 

Archives of Razi Institute
Volume 76, Issue 5
November and December 2021
Pages 1363-1370

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