A comparative study was performed to evaluate best practice culture media and enrichment broths for recovering Salmonella species from human stool samples. A total of 1297 human stools were collected and processed in this study. Evaluation of agar media was carried out by direct plating (DP), 1096 stool samples were inoculated on Modified Semisolid Rappaport-Vassiliadis (MSRV), Xylose-Lysine-Deoxycolate (XLD), MacConkey (MAC), and Hektoen Enteric (HE) agars. Evaluation of enrichment broths were carried out by enrichment all 1297 stool samples in Selenite broth (SB), Rappaport-Vassiliadis (RV) broth, and Buffered Peptone Water (BPW), followed by plating on MSRV, MAC, and HE agars. A total of 102 Salmonella-positive stools by DP, 85.3% (87/102) were recovered utilizing MSRV while recovery from XLD, MAC, and HE agars were 34.3% (35/102), 34.3% (35/102), and 29.4% (30/102) respectively. A total 299/1297 stools samples were Salmonella-positive on at least one plating medium after enrichment procedure were 77.3% (177/299) for SB, 86.0% (197/299) and 78.6% (180/299) for RV and BPW respectively. All Salmonella isolated in this study was nontyphi Salmonella. Presently, the data suggest that the use of MSRV over MAC, HE, and XLD agars for isolation nontyphi Salmonella species from human stools is more efficacious. Additionally, use of MSRV in combination with MAC and HE agars following enrichment in RV broth enhances recovery of nontyphi Salmonella species. However, RV broth is inhibitory to typhi Salmonella, thus use of MSRV medium in combination with MAC, HE or XLD agars in direct plating following enrichment in non-selective BPW is an alternate method for recovery of both typhi and nontyphi Salmonella species contaminated in human stool samples.
References
[1]
Voetsch, A.C., Van Gilder, T.J., Angulo, F.J., et al. (2004) Foodnet Estimate of the Burden of Illness Caused by Non-typhoidal Salmonella Infections in the United States. Clinical Infectious Diseases, 38, 127-134. http://dx.doi.org/10.1086/381578
[2]
Varavithaya, W., Vathanophas, K., Bodhidatta, L., et al. (1990) Importance of Salmonellae and Campylobacter jejuni in the Etiology of Diarrheal Disease Among Children Less than 5 Years of Age in a Community in Bangkok, Thailand. Journal of Clinical Microbiology, 2507-2510.
[3]
Bodhidatta, L., Vithayasai, N., Eimpokalarp, B., Pitarangsi, C., Serichantalergs, O. and Isenbarger, D.W. (2002) Bacterial Enteric Pathogens in Children with Acute Dysentery in Thailand: Increasing Importance of Quinolone-Resistant Campylobacter. The Southeast Asian Journal of Tropical Medicine and Public Health, 33, 752-757.
[4]
Devenish, J.A., Ciebin, B.W. and Brodsky, M.H. (1986) Novobiocin-Brilliant Green-Glucose Agar: New Medium for Isolation of Salmonellae. Applied and Environmental Microbiology, 52, 539-545.
[5]
Rambach, A. (1990) New Plate Medium for Facilitated Differentiation of Salmonella spp. from Proteus spp. and Other Enteric Bacteria. Journal of Applied Environmental and Microbiology, 56, 301-303.
[6]
Ruiz, J., Nunez, M.L., Lorente, I., Perez, J., Simarro, E. and Gomez, J. (1996) Performance of Six Culture Media for Isolation of Salmonella Species from Stool Samples. European Journal of Clinical Microbiology & Infectious Diseases, 15, 922-926. http://dx.doi.org/10.1007/BF01690509
[7]
Nye, K.J., Fallon, D., Frodsham, D., et al. (2002) An Evaluation of the Performance of XLD, DCA, MLCB, and ABC Agars as Direct Plating Media for Isolation of Salmonella Enteric from Faeces. Journal of Clinical Pathology, 55, 286-288. http://dx.doi.org/10.1136/jcp.55.4.286
[8]
Jones, R.E. and Handley, W.R.C. (1945) A Selective Medium for the Isolation of Salmonella Bacilli from Heavily Contaminated Material. Monthly Bulletin of the Ministry of Health and the Public Health Laboratory Service, 4, 107-111.
[9]
Ino, J. and Graber, C.D. (1955) Recovery of Salmonella from Contaminated Cultures. US Armed Forces Medical Journal, 6, 586-587.
[10]
Stuart, P.F. and Pivnick, H. (1965) Isolation of Salmonellae by Selective Motility Systems. Applied Microbiology, 13, 365-372.
[11]
Harper, J. and Shortridge, K.F. (1969) A Selective Motility Medium for Routine Isolation of Salmonella. Journal of Hygiene (Lond), 67, 181-186. http://dx.doi.org/10.1017/S0022172400041589
[12]
Rappaport, F., Konforti, N. and Navon, B. (1956) A New Enrichment Medium for Certain Salmonellae. Journal of Clinical Pathology, 9, 261-266. http://dx.doi.org/10.1136/jcp.9.3.261
[13]
Vassiliadis, P., Trichopoulos, D., Pateraki, E. and Papaiconomou, N. (1978) Isolation of Salmonella from Minced Meat by the Use of a New Procedure of Enrichment. Zentralblatt fur Bakteriologie Originale B. Hygiene, Preventive Medezin, 166, 81-86.
[14]
De Smedt, J.M., Bolderdijk, R.F., Rappold, H. and Lautenschlaeger, D. (1986) Rapid Salmonella Detection in Food by Motility Enrichment on a Modified Semi-Solid Rappaport-Vassiliadis Medium. Journal of Food Protection, 49, 510-514.
[15]
De Smedt, J.M. and Bolderdijk, R.F. (1987) Dynamics of Salmonella Isolation with Modified Semi-Solid Rappaport Vassiliadis Medium. Journal of Food Protection, 50, 658-661.
[16]
Aspinall, S.T., Hindle, M.A. and Hutchinson, D.N. (1992) Improved Isolation of Salmonellae from Faeces Using a Semisolid Rappaport-Vassiliadis Medium. European Journal of Clinical Microbiology & Infectious Diseases, 11, 936-939. http://dx.doi.org/10.1007/BF01962379
[17]
Dusch, H. and Altwegg, M. (1995) Evaluation of Five New Plating Media for Isolation of Salmonella Species. Journal of Clinical Microbiology, 33, 802-804.
[18]
Ruiz, J., Lorente, I., Perez, J., Simarro, E. and Martinez, L. (1998) Evaluation of Methods for Isolation of Salmonella Species Using Modified Semisolid Rappaport-Vassiliadis Medium and Salmonella-Shigella Agar. European Journal of Clinical Microbiology & Infectious Diseases, 17, 791-793. http://dx.doi.org/10.1007/s100960050188
[19]
Goossens, H., Wauters, G., De Boeck, M., Janssens, M. and Butzler, J.P. (1984) Semisolid Selective-Motility Enrichment Medium for Isolation of Salmonellae from Fecal Specimens. Journal of Clinical Microbiology, 19, 940-941.