Salmonella infection is one of the major global public health hazards and remains as an economic burden to both developed and developing countries through the costs associated with trade banning, as well as surveillance, prevention, and treatment of the disease. Confirmatory and rapid identification of Salmonella in feed and foods of animal origin is crucial for mitigating the associated burdens. The conventional methods for isolation and identification of Salmonella species are based on culture and biochemical tests and are very time-consuming requiring 10-11 days. Hence, these drawbacks of the conventional methods warrant a rapid method for confirmatory identification of Salmonella in feed and foods of animal origin. Therefore, a method based on matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) was developed and verified for its reliable use in the laboratory. For verification, Salmonella typhimurium (ATCC 14028) was used as the target bacteria. E. coli (ATCC 25922) and Staphylococcus aureus (ATCC 25923) were used as non-target bacteria. Salmonella typhimurium (ATCC 14028) produced black colonies with metallic sheen on BS agars, black colonies on XLD agars, and black-centered blue colonies on HE agars. MALDI TOF MS was found very efficient for confirmatory identification of Salmonella bacteria at the genus level without performing biochemical tests. Confirmatory identification of Salmonella could be made within four to five days after the commencement of the test. The expected limit of detection (eLOD50) of Salmonella in chicken and beef was found < 1.7 ± 0, in milk 1.2 ± 0.2, in eggs < 1.2 ± 0.2, and in feed pellet < 1.6 ± 0.2 CFU/test portion. The accuracy, sensitivity, and specificity of the method were found 98%, 96%, and 100% respectively. No significant effects (p = 0.400) of sample matrices were found on test results carried out by the developed method. Likewise, in terms of ruggedness, no significant effects of analyst change (p = 0.787), incubator change (p = 0.787), and day change (p = 0.242) were found in the test results. The method was found robust and could be used in the laboratory for rapid and confirmatory identification of Salmonella in feeds, and foods of animal origin.
| Published in | International Journal of Microbiology and Biotechnology (Volume 7, Issue 2) |
| DOI | 10.11648/j.ijmb.20220702.16 |
| Page(s) | 84-92 |
| Creative Commons |
This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited. |
| Copyright |
Copyright © The Author(s), 2024. Published by Science Publishing Group |
Salmonella, Identification Method, Verification, MALDI TOF MS
| [1] | McSorley, S. J. (2014) Immunity to intestinal pathogens: lessons learned from Salmonella. Immunological Reviews. 260 (1): 168-182. |
| [2] | Chlebicz, A. and Śliżewska, K. (2018) Campylobacteriosis, salmonellosis, yersiniosis, and listeriosis as zoonotic foodborne diseases: a review. International Journal of Environmental Research and Public Health. 15 (5): 863. |
| [3] | Rönnqvist, M., Välttilä, V., Ranta, J., and Tuominen, P. (2018) Salmonella risk to consumers via pork is related to the Salmonella prevalence in pig feed. Food microbiology. 71: 93-97. |
| [4] | Eng, S.-K., Pusparajah, P., Ab Mutalib, N.-S., Ser, H.-L., Chan, K.-G., and Lee, L.-H. (2015) Salmonella: a review on pathogenesis, epidemiology and antibiotic resistance. Frontiers in Life Science. 8 (3): 284-293. |
| [5] | Majowicz, S. E., Musto, J., Scallan, E., Angulo, F. J., Kirk, M., O'Brien, S. J., Jones, T. F., Fazil, A., Hoekstra, R. M., and Studies, I. C. o. E. D. B. o. I. (2010) The global burden of nontyphoidal Salmonella gastroenteritis. Clinical Infectious Diseases. 50 (6): 882-889. |
| [6] | Ferrari, R. G., Rosario, D. K., Cunha-Neto, A., Mano, S. B., Figueiredo, E. E., and Conte-Junior, C. A. (2019) Worldwide epidemiology of Salmonella serovars in animal-based foods: a meta-analysis. Applied and Environmental Microbiology. 85 (14): e00591-00519. |
| [7] | Paniel, N. and Noguer, T. (2019) Detection of Salmonella in food matrices, from conventional methods to recent aptamer-sensing technologies. Foods. 8 (9): 371. |
| [8] | Whiley, H. and Ross, K. (2015) Salmonella and eggs: from production to plate. International Journal of Environmental Research and Public Health. 12 (3): 2543-2556. |
| [9] | Ncoko, P., Jaja, I. F., and Oguttu, J. W. (2020) Microbiological quality of beef, mutton, and water from different abattoirs in the Eastern Cape Province, South Africa. Veterinary world. 13 (7): 1363. |
| [10] | de Boer, E. and Beumer, R. R. (1999) Methodology for detection and typing of foodborne microorganisms. International Journal of Food Microbiology. 50 (1-2): 119-130. |
| [11] | Andrews, W. H., Jacobson, A., and Hammack, T., Bacteriological Analytical Manual Chapter 5: Salmonella. 2021, AOAC International Gaithersburg. p. 5.01-05.020. https://www.fda.gov/food/laboratory-methods-food/bam-chapter-025-salmonella [retrieved on 2/11/2021]. |
| [12] | ISO 6579-1: 2017, (2017) Microbiology of the food chain — Horizontal method for the detection, enumeration and serotyping of Salmonella — Part 1: Detection of Salmonella spp. International Organization for Standardization. ISO Central Secretariat, Chemin de Blandonnet 8, CP 401-1214 Vernier, Geneva, Switzerland: https://www.iso.org/standard/56712.html [retrieved on 5/11/2021]. |
| [13] | Kang, L., Li, N., Li, P., Zhou, Y., Gao, S., Gao, H., Xin, W., and Wang, J. (2017) MALDI-TOF mass spectrometry provides high accuracy in identification of Salmonella at species level but is limited to type or subtype Salmonella serovars. European journal of mass spectrometry. 23 (2): 70-82. |
| [14] | Pavlovic, M., Huber, I., Konrad, R., and Busch, U. (2013) Application of MALDI-TOF MS for the identification of food borne bacteria. The open microbiology journal. 7: 135. |
| [15] | Torres-Sangiao, E., Leal Rodriguez, C., and García-Riestra, C. (2021) Application and Perspectives of MALDI–TOF Mass Spectrometry in Clinical Microbiology Laboratories. Microorganisms. 9 (7): 1539. |
| [16] | Angeletti, S. (2017) Matrix assisted laser desorption time of flight mass spectrometry (MALDI-TOF MS) in clinical microbiology. Journal of Microbiological Methods. 138: 20-29. |
| [17] | Gaudreau, A., Labrie, J., Goetz, C., Dufour, S., and Jacques, M. (2018) Evaluation of MALDI-TOF mass spectrometry for the identification of bacteria growing as biofilms. Journal of Microbiological Methods. 145: 79-81. |
| [18] | Bastin, B., Bird, P., Benzinger, M. J., Crowley, E., Agin, J., Goins, D., Sohier, D., Timke, M., Shi, G., and Kostrzewa, M. (2018) Confirmation and Identification of Salmonella spp., Cronobacter spp., and Other Gram-Negative Organisms by the Bruker MALDI Biotyper Method: Collaborative Study, First Action 2017.09. Journal of AOAC International. 101 (5): 1593-1609. |
| [19] | 16140-3: 2017, I. (2017) Microbiology of the food chain — Method validation — Part 3: Protocol for the verification of reference methods and validated alternative methods in a single laboratory. International Organization for Standardization. ISO Central Secretariat, Chemin de Blandonnet 8, CP 401-1214 Vernier, Geneva, Switzerland: https://www.iso.org/standard/66324.html [retrieved on 66305/66311/62020]. |
| [20] | National Association of Testing Authorities, Australia. (2012) Guidelines for the validation and verification of quantitative and qualitative test methods. http://www.demarcheiso17025.com/document/Guidelines%17020for%17020the%17020validation%17020and%17020verification%17020of%17020quantitative%17020and%17020qualitative%17020test%17020methods.pdf [retrieved on 5/11/2020]. |
| [21] | Simione, F. P. (2009) Thermo scientific nalgene and nunc cryopreservation guide. Thermo Fisher Scientific Incorporation, New York. https://tools.thermofisher.com/content/sfs/brochures/D17044.pdf [retrieved on 5/11/2020]. |
| [22] | Hantash, T., Nauta, M., Al-Sabi, M. N., Al-ali, W. Q., and Vigre, H. (2020) The “Rapid’Salmonella” Method: Estimation of the Limit of Detection for Salmonella Strains Typhimurium and Enteritidis Isolated from Frozen Poultry Meat. Modern Applied Science. 14 (12): 43-51. |
| [23] | Maturin, L. and Peeler, J. (2001) Bacteriological Analytical Manual (BAM) Chapter 3: Aerobic plate count. Bacteriological analytical manual (BAM), US Food and Drug Administration. https://www.fda.gov/food/laboratory-methods-food/bam-chapter-3-aerobic-plate-count. [retrieved on 15/3/2021]. |
| [24] | Shell, W. S., Sayed, M. L., Allah, F. M. G., Gamal, F. E. M., Khedr, A. A., Samy, A., and Ali, A. H. M. (2017) Matrix-assisted laser desorption-ionization-time-of-flight mass spectrometry as a reliable proteomic method for characterization of Escherichia coli and Salmonella isolates. Veterinary world. 10 (9): 1083. |
| [25] | Baratloo, A., Hosseini, M., Negida, A., and El Ashal, G. (2015) Part 1: simple definition and calculation of accuracy, sensitivity and specificity. Emergency. 3 (2): 48-49. |
| [26] | Dhama, K., Rajagunalan, S., Chakraborty, S., Verma, A., Kumar, A., Tiwari, R., and Kapoor, S.(2013) Food-borne pathogens of animal origin-diagnosis, prevention, control and their zoonotic significance: a review. Pakistan journal of biological sciences: PJBS. 16 (20): 1076-1085. |
APA Style
Md. Al-Amin, Md. Mizanur Rahman, Marufa Aktar, Md. Mostofa Kamal. (2022). Development and Verification of a MALDI-TOF MS-Based Method for Rapid and Confirmatory Identification of Salmonella in Feed and Foods. International Journal of Microbiology and Biotechnology, 7(2), 84-92. https://doi.org/10.11648/j.ijmb.20220702.16
ACS Style
Md. Al-Amin; Md. Mizanur Rahman; Marufa Aktar; Md. Mostofa Kamal. Development and Verification of a MALDI-TOF MS-Based Method for Rapid and Confirmatory Identification of Salmonella in Feed and Foods. Int. J. Microbiol. Biotechnol. 2022, 7(2), 84-92. doi: 10.11648/j.ijmb.20220702.16
AMA Style
Md. Al-Amin, Md. Mizanur Rahman, Marufa Aktar, Md. Mostofa Kamal. Development and Verification of a MALDI-TOF MS-Based Method for Rapid and Confirmatory Identification of Salmonella in Feed and Foods. Int J Microbiol Biotechnol. 2022;7(2):84-92. doi: 10.11648/j.ijmb.20220702.16
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Download@article{10.11648/j.ijmb.20220702.16,
author = {Md. Al-Amin and Md. Mizanur Rahman and Marufa Aktar and Md. Mostofa Kamal},
title = {Development and Verification of a MALDI-TOF MS-Based Method for Rapid and Confirmatory Identification of Salmonella in Feed and Foods},
journal = {International Journal of Microbiology and Biotechnology},
volume = {7},
number = {2},
pages = {84-92},
doi = {10.11648/j.ijmb.20220702.16},
url = {https://doi.org/10.11648/j.ijmb.20220702.16},
eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijmb.20220702.16},
abstract = {Salmonella infection is one of the major global public health hazards and remains as an economic burden to both developed and developing countries through the costs associated with trade banning, as well as surveillance, prevention, and treatment of the disease. Confirmatory and rapid identification of Salmonella in feed and foods of animal origin is crucial for mitigating the associated burdens. The conventional methods for isolation and identification of Salmonella species are based on culture and biochemical tests and are very time-consuming requiring 10-11 days. Hence, these drawbacks of the conventional methods warrant a rapid method for confirmatory identification of Salmonella in feed and foods of animal origin. Therefore, a method based on matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) was developed and verified for its reliable use in the laboratory. For verification, Salmonella typhimurium (ATCC 14028) was used as the target bacteria. E. coli (ATCC 25922) and Staphylococcus aureus (ATCC 25923) were used as non-target bacteria. Salmonella typhimurium (ATCC 14028) produced black colonies with metallic sheen on BS agars, black colonies on XLD agars, and black-centered blue colonies on HE agars. MALDI TOF MS was found very efficient for confirmatory identification of Salmonella bacteria at the genus level without performing biochemical tests. Confirmatory identification of Salmonella could be made within four to five days after the commencement of the test. The expected limit of detection (eLOD50) of Salmonella in chicken and beef was found p = 0.400) of sample matrices were found on test results carried out by the developed method. Likewise, in terms of ruggedness, no significant effects of analyst change (p = 0.787), incubator change (p = 0.787), and day change (p = 0.242) were found in the test results. The method was found robust and could be used in the laboratory for rapid and confirmatory identification of Salmonella in feeds, and foods of animal origin.},
year = {2022}
}
TY - JOUR T1 - Development and Verification of a MALDI-TOF MS-Based Method for Rapid and Confirmatory Identification of Salmonella in Feed and Foods AU - Md. Al-Amin AU - Md. Mizanur Rahman AU - Marufa Aktar AU - Md. Mostofa Kamal Y1 - 2022/06/09 PY - 2022 N1 - https://doi.org/10.11648/j.ijmb.20220702.16 DO - 10.11648/j.ijmb.20220702.16 T2 - International Journal of Microbiology and Biotechnology JF - International Journal of Microbiology and Biotechnology JO - International Journal of Microbiology and Biotechnology SP - 84 EP - 92 PB - Science Publishing Group SN - 2578-9686 UR - https://doi.org/10.11648/j.ijmb.20220702.16 AB - Salmonella infection is one of the major global public health hazards and remains as an economic burden to both developed and developing countries through the costs associated with trade banning, as well as surveillance, prevention, and treatment of the disease. Confirmatory and rapid identification of Salmonella in feed and foods of animal origin is crucial for mitigating the associated burdens. The conventional methods for isolation and identification of Salmonella species are based on culture and biochemical tests and are very time-consuming requiring 10-11 days. Hence, these drawbacks of the conventional methods warrant a rapid method for confirmatory identification of Salmonella in feed and foods of animal origin. Therefore, a method based on matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) was developed and verified for its reliable use in the laboratory. For verification, Salmonella typhimurium (ATCC 14028) was used as the target bacteria. E. coli (ATCC 25922) and Staphylococcus aureus (ATCC 25923) were used as non-target bacteria. Salmonella typhimurium (ATCC 14028) produced black colonies with metallic sheen on BS agars, black colonies on XLD agars, and black-centered blue colonies on HE agars. MALDI TOF MS was found very efficient for confirmatory identification of Salmonella bacteria at the genus level without performing biochemical tests. Confirmatory identification of Salmonella could be made within four to five days after the commencement of the test. The expected limit of detection (eLOD50) of Salmonella in chicken and beef was found p = 0.400) of sample matrices were found on test results carried out by the developed method. Likewise, in terms of ruggedness, no significant effects of analyst change (p = 0.787), incubator change (p = 0.787), and day change (p = 0.242) were found in the test results. The method was found robust and could be used in the laboratory for rapid and confirmatory identification of Salmonella in feeds, and foods of animal origin. VL - 7 IS - 2 ER -
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