Submit Manuscript

International Journal of Poultry Science

Year: 2011 | Volume: 10 | Issue: 2 | Page No.: 93-97
Facebook Twitter Digg Reddit Linkedin StumbleUpon E-mail
Research Article

Detection of Salmonella and Escherichia coli in Egg Shell and Egg Content from Different Housing Systems for Laying Hens

Wiriya Loongyai, Benjamaporn Wiriya and Naruemol Sangsawang

ABSTRACT

Polymerase Chain Reaction (PCR) assay and conventional microbiological methods were used to detect bacterial contamination of egg shells and egg content in different commercial housing systems, the open house and evaporative cooling systems. A PCR assay was developed for direct detection using a set of primers specific for the invasion by invA gene for Salmonella and uidA gene, encoding β-glucuronidase for Escherichia coli. PCR detected the presence of Salmonella in 2 (5%) samples of egg shells from the evaporative cooling system, while conventional cultural methods detected no Salmonella from the same samples. No Salmonella was detected from the open house system. The frequency of PCR positive samples for E. coli was 35% (16/40) isolated from the open house system and 42% (16/38) from the evaporative cooling system. Two samples screened for E. coli in egg content was also detected by PCR. In comparison, detection of E. coli by culture, done simultaneously with PCR, was 17.5% (7/40) sensitive in the open house system and 31.5% (12/38) sensitive in the evaporative cooling system. Results indicated that this PCR procedure is a sensitive method for Salmonella and E. coli detection. The contamination of Salmonella and E. coli was found at a high frequency in the evaporative cooling system compared with the open house system.
PDF References Citation

Search

REFERENCES

  1. Akhtar, F., H. Afzal, M. Ashfaque and Z. Nasim, 1982. Egg-shell contaminations of human importance with reference to Salmonellae. J. Pak. Vet., 2: 10-11.
  2. Brasher, C., G. Panicker and A.K. Bej, 2002. Evaluation of PCR amplification based detection of heat-killed Escherichia coli and cell-free DNA in shellfish. Mol. Biol. Today, 3: 85-90.
    Direct Link
  3. Bulte, M. and P. Jakob, 1995. The use of a PCR-generated invA probe for the detection of Salmonella sp. in artificially and naturally contaminated foods. Int. J. Food Microbial., 26: 335-344.
    CrossRef
  4. Bonetta, S., E. Borelli, S. Bonetta, O. Conio, F. Palumbo and E. Carraro, 2010. Development of a PCR protocol for the detection of Escherichia coli O157:H7 and Salmonella sp. in surface water. Environ. Monit. Assess.
  5. Cebula, T.A., W.L. Payne and P. Feng, 1995. Simultaneous identification of strains of Escherichia coli serotype O157: H7 and their Shiga-like toxin type by mismatch amplification mutation assay-multiplex PCR. J. Clin. Microbiol., 33: 248-250.
    Direct Link
  6. Daly, P., T. Collier and S. Doyle, 2002. PCR-ELISA detection of Escherichia coli in milk. Lett. Applied Microbiol., 34: 222-226.
    CrossRefDirect Link
  7. Darwin, K.H. and V.L. Miller, 1999. Molecular basis of the interaction of Salmonella with the intestinal mucosa. Clin. Microbiol. Rev., 12: 405-428.
    PubMedDirect Link
  8. De Reu, K., K. Grijspeerdt, W. Messens, M. Heyndrickx, M. Uyttendaele, J. Debevere and L. Herman, 2006. Eggshell factors influencing eggshell penetration and whole egg contamination by different bacteria, including Salmonella enteritidis. Int. J. Food Microbiol., 112: 253-260.
    CrossRefDirect Link
  9. De Reu, K., K. Grijspeerdt, M. Heyndrickx, M. Uyttendaele and L. Herman, 2006. Bacterial eggshell contamination in the egg production chain and in different housing systems. Br. Poult. Sci., 47: 163-172.
  10. Desmarchelier, P.M., S.S. Bilge, N. Fegan, L. Mills, J.C. Jr. Vary and P.I. Tarr, 1998. A PCR specific for Escherichia coli O157 based on the rfbE locus encoding O157 lipopolysaccharide. J. Clin. Microbiol., 36: 1801-1804.
    Direct Link
  11. Doran, J.L., S.K. Collinson, J. Burian, G. Sarlos and E.C.D. Todd et al., 1993. DNA-based diagnostic test for Salmonella species targeting agfA, the structural gene for thin aggregative fimbriae. J. Clin. Microbiol., 31: 2263-2273.
    Direct Link
  12. Doyle, M.P. and J.L. Schoeni, 1987. Isolation of Escherichia coli O157: H7 from retail fresh meats and poultry. Applied Environ. Microbiol., 53: 2394-2396.
    Direct Link
  13. Ferretti, R., L. Mannazzu, L. Cocolin, G. Comi and F. Clementi, 2001. Twelve-hours PCR-based method for detection of Salmonella sp. in Food. Applied Environ. Microbiol., 67: 977-978.
    CrossRef
  14. Gannon, V.P.J., M. Rashed, R.K. King and E.J.G. Thomas, 1993. Detection and characterization of the eae gene of shiga-like toxin-producing Escherichia coli using polymerase chain reaction. J. Clin. Microbiol., 31: 1268-1274.
    Direct Link
  15. Gebreyes, W.A., P.R. Davies, W.E.M. Morrow, J.A. Funk and C. Altier, 2000. Antimicrobial resistance of Salmonella isolates from swine. J. Clin. Microbiol., 38: 4633-4636.
    Direct Link
  16. Guo, L., J. Killefer, P.B. Kenney and J.D. Mick-Morris, 1999. Use of arbitrarily primed polymerase chain reaction to study Salmonella ecology in a turkey production environment. Poult. Sci., 78: 24-31.
    PubMed
  17. Haines, R.B., 1938. Observation on the bacterial flora of the hen's egg with a discription of new species of Proteus and Pseudomonas causing rot in eggs. J. Hyg., 38: 338-355.
    Direct Link
  18. Harry, E.G., 1963. The relationship between egg spoilage and the environment of the egg when laid. Br. Poult. Sci., 4: 91-100.
    Direct Link
  19. Heininger, A., M. Binder, S. Schmidt, K. Unertl, Botzenhart and G. Doring, 1999. PCR and blood culture for detection of Escherichia coli bacteremia in rats. J. Clin. Microbiol., 37: 2479-2482.
    Direct Link
  20. Hashimoto, Y., Y. Itho, Y. Fujinaga, A.Q. Khan and F. Sultana et al., 1995. Development of nested PCR based on the ViaB sequence to detect Salmonella typhi. J. Clin. Microbiol., 33: 775-777.
    Direct Link
  21. Heijnen, L. and G. Medema, 2009. Method for rapid detection of viable Escherichia coli in water using real-time NASBA. Water Res., 43: 3124-3132.
    CrossRef
  22. Holland, J.L., L. Louie, A.E. Simor and M. Louie, 2000. PCR detection of Escherichia coli O157: H7 directly from stools: Evaluation of commercial extraction methods for purifying fecal DNA. J. Clin. Microbiol., 38: 4108-4113.
    Direct Link
  23. Iida, K., A. Abe, H. Matsui, H. Danbara, S. Wakayama and K. Kawahara, 1993. Rapid and sensitive method for detection of Salmonella strains using a combination of polymerase chain reaction and reverse dot-blot hybridization. FEMS Microbiol. Lett., 114: 167-172.
    CrossRef
  24. Jones, F.T., D.V. Rives and J.B. Carey, 1995. Salmonella contamination in commercial eggs and an egg production facility. Poult. Sci., 74: 753-757.
    PubMed
  25. Jothikumar, N. and M.W. Griffiths, 2002. Rapid detection of Escherichia coli O157:H7 with multiplex real-time PCR assays. Applied Environ. Microbiol., 68: 3169-3171.
    Direct Link
  26. Mahon, J. and A. Lax, 1993. A quantitative polymerase chain reaction method for the detection in avian faeces of Salmonella carrying the spvR gene. Epidemiol. Infect., 111: 455-464.
    Direct Link
  27. Mahon, J., C.K. Murphy, P.W. Jones and P.A. Barrow, 1994. Comparison of multiplex PCR and standard bacteriological methods of detecting Salmonella on chicken skin. Lett. Applied Microbiol., 19: 169-172.
    CrossRef
  28. Yes, F.J. and M.A. Takeballi, 1983. Microbial contamination of the hen's egg: A review. J. Food Prot., 46: 1092-1098.
    Direct Link
  29. Oberst, R.D., M.P. Hays, L.K. Bohra, R.K. Phebus and J.M. Sargeant, 2003. Detection of Escherichia coli O157:H7 in cattle feces using a polymerase chain reaction based fluorogenic 59 nuclease (TaqMant) detection assay after secondary enrichment. J. Vet. Diagn. Invest., 15: 543-552.
    Direct Link
  30. Paton, A.W. and J.C. Paton, 1998. Detection and characterization of Shiga toxigenic Escherichia coli by using multiplex PCR assays for stx1, stx2, eaeA, enterohemorrhagic E. coli hlyA, rfbO111 and rfbO157. J. Clin. Microbiol., 36: 598-602.
    PubMedDirect Link
  31. Rahn, K., S.A. De Grandis, R.C. Clarke, S.A. McEwen and J.E. Galan et al., 1992. Amplification of an invA gene sequence of Salmonella typhimurium by polymerase chain reaction as a specific method of detection of Salmonella. Mol. Cell. Probes., 6: 271-279.
    CrossRefPubMedDirect Link
  32. Rahman, M.M., Z.U.M. Khan, A.N.M. Rahman and M.S. Islam, 2006. Recovery of Salmonella from incubated eggs, newly hatched chicks and contaminated environments. J. Anim. Vet. Adv., 5: 598-601.
    Direct Link
  33. Rajashekara, G., E. Harverly, D.A. Halvorson, K.E. Ferris, D.C. Lauer and K.V. Nagaraja, 2000. Multidrug resistant Salmonella typhimurium DT104 in poultry. J. Food Prot., 63: 155-161.
    Direct Link
  34. Robles, M.A.G., A.M. Loredo, G.A. Ojeda, J.A.O. Garcia, I.O. Martinez, L.H.M. Ramos and P. Fratamico, 2009. PCR detection and microbiological isolation of Salmonella sp. from fresh beef and cantaloupes. J. Food. Sci., 74: M37-40.
    Direct Link
  35. Rozila, A., A.W. Hartini, D.Y. Tan and S.K. Wee, 2007. Rapid molecular detection of Salmonella isolated from poultry farm. Proceedings of the 19th Veterinary Association Malaysia Congress, Aug. 3-5, Malaysia, pp: 201-203.
  36. Sahilah, A.M., L.Y.Y. Audrey, S.L. Ong, W.N. Wan Sakeenah and S. Safiyyah et al., 2010. DNA profiling among egg and beef meat isolates of Escherichia coli by Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR) and Random Amplified Polymorphic DNA-PCR (RAPD-PCR). Int. Food Res. J., 17: 853-866.
    Direct Link
  37. Salehi, T.Z., M. Mahzounieh and A. Saeedzadeh, 2005. Detection of InvA gene in isolated Salmonella from broilers by PCR method. Int. J. Poult. Sci., 4: 557-559.
    CrossRefDirect Link
  38. Santos, L.R., V.P. Nascimento, S.D. Oliveira, M.L. Flores and A.P. Pontes et al., 2001. Polymerase Chain Reaction (PCR) for the detection of Salmonella in artificially inoculated chicken meat. Rev. Inst. Med. Trop. Sao. Paul., 43: 247-250.
    Direct Link
  39. Schneder, A., C. Gronewald, M. Fandke, B. Kurth, S. Barkowski and K. Berghof-Ager, 2002. Real-time detection of the genus Salmonella with the light cycler system. Biochemica, 4: 19-21.
    Direct Link
  40. Schoeni, J.L., K.A. Glass, J.L. McDermott and A.C.L. Wong, 1995. Growth and penetration of Salmonella enteritidis, Salmonella heidelberg and Salmonella typhimurium in eggs. Int. J. Food Microbiol., 24: 385-396.
    CrossRef
  41. Schrank, I.S., M.A.Z. Mores, J.L.A. Costa, A.P.G. Frazzon and R. Sonicini et al., 2001. Influence of enrichment media and application of a PCR based method to detect Salmonella in poultry industry products and clinical samples. Vet. Microbiol., 82: 45-53.
    Direct Link
  42. Sharma, V.K. and S.A. Carlson, 2000. Simultaneous detection of Salmonella strains and Escherichia coli O157:H7 with fluorogenic PCR and single-enrichment-broth culture. Applied Environ. Microbiol., 66: 5472-5476.
    Direct Link
  43. Touron, A., T. Berthe, B. Pawlak and F. Petit, 2005. Detection of Salmonella in environmental water and sediment by a nested-multiplex polymerase chain reaction assay. Res. Microbiol., 156: 541-553.
    CrossRefDirect Link
  44. Trampel, D.W., Y. Wannemuehler and L.K. Nolan, 2007. Characterization of Escherichia coli isolates from Peritonitis lesions in commercial laying hens. Avian Dis., 51: 840-844.
    Direct Link
  45. Wang, H., B.W. Blais and H. Yamazaki, 1995. Rapid confirmation of polymyxin-cloth enzyme immunoassay for group D salmonellae including Salmonella enteritidis in eggs by polymerase chain reaction. Food Control, 6: 205-209.
    CrossRef

Search

Leave a Comment

Your email address will not be published. Required fields are marked *