Hostname: page-component-848d4c4894-xm8r8 Total loading time: 0 Render date: 2024-07-01T07:59:24.500Z Has data issue: false hasContentIssue false

Quantitation of viable Coxiella burnetii in milk products using a liquid medium-based MPN-PCR assay

Published online by Cambridge University Press:  28 August 2018

Manman Shi
Affiliation:
Illinois Institute of Technology, Institute for Food Science & Health, Bedford Park, IL 60501, USA
Cheng Zhang
Affiliation:
Illinois Institute of Technology, Institute for Food Science & Health, Bedford Park, IL 60501, USA
Diana Stewart*
Affiliation:
Division of Food Processing Science & Technology, US Food and Drug Administration, Bedford Park, IL 60501, USA
*
*For correspondence; e-mail: diana.stewart@fda.hhs.gov

Abstract

This Technical Research Communication describes a new method by which thermally treated Coxiella in milk products may be grown in a liquid growth medium and quantitated using an MPN-PCR assay. Coxiella is generally not used in studies on thermal and non-thermal processing of milk due to the need for specialized and highly laborious techniques such as animal assays and tissue culture for determining viability. Recently, a liquid growth medium (ACCM-2) and modified atmosphere were used to grow Coxiella from pure cultures, infected mouse tissues, and clinical samples, however, the ability to grow Coxiella from a food such as milk has not been shown. The potential ability to enrich Coxiella directly from contaminated milk presents a new avenue for conducting pasteurization research in which the viability of heat-treated or injured cultures could be more easily determined through direct enrichment of Coxiella in ACCM-2. ACCM-2 medium allowed enrichment of Coxiella from bovine whole milk and cream, whole goat, and whole camel milks but not whole water buffalo milk. Enrichment was possible from whole bovine milk containing as few as 6 Coxiella ge/ml of milk. The applicability of this ACCM-2 enrichment method was shown when using an MPN-PCR assay to quantitate the number of viable Coxiella remaining in whole bovine milk after 64 °C thermal treatment for up to 10 min.

Type
Research Article
Copyright
Copyright © Hannah Dairy Research Foundation 2018 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Blodgett 2010 FDA Bacteriological Analytical Manual Appendix 2 Most Probably Number from Serial Dilutions Available at: http://www.fda.gov/Food/FoodScienceResearch/LaboratoryMethods/ucm109656.htm (Accessed 29 June 2017)Google Scholar
Boden, K, Wolf, K, Hermann, B & Frangoulidis, B 2015 First isolation of Coxiella burnetii from clinical material by cell-free medium (ACCM2). European Journal of Microbiology & Infectious Diseases 34 10171022Google Scholar
Bradshaw, JG, Peller, JT, Corwin, JJ, Barnett, JE & Twedt, RM 1987 Thermal resistance of disease-associated Salmonella typhimurium in milk. Journal of Food Protection 50 9596Google Scholar
Bunning, VK, Donnelly, CW, Peeler, JT, Briggs, EH, Bradshaw, JG, Crawford, RG, Beliveau, CM & Tierney, JT 1988 Thermal inactivation of Listeria monocytogenes within bovine milk phagocytes. Applied & Environmental Microbiology 54 364370Google Scholar
Cerf, O & Condron, R 2006 Coxiella burnetii and milk pasteurization: an early application of the precautionary principle? Epidemiology & Infection 134 946951Google Scholar
D'Aoust, J-Y, Park, CE, Szabo, RA & Todd, ECD 1988 Thermal inactivation of Campylobacter species, Yersinia enterocolitica, and hemorrhagic Escherichia coli O157:H7 in fluid milk. Journal of Dairy Science 71 32303236Google Scholar
Donnelly, CW & Briggs, EH 1986 Psychrotrophic growth and thermal inactivation of Listeria monocytogenes as a function of milk composition. Journal of Food Protection 49 994998Google Scholar
Doyle, MP, Glass, KA, Beery, JT, Garcia, GA, Pollard, DJ & Schultz, RD 1987 Survival of Listeria monocytogenes in milk during high-temperature, short-time pasteurization. Applied & Environmental Microbiology 53 14331438Google Scholar
Enright, JB 1961 The pasteurization of cream, chocolate milk and ice cream mixes containing the organisms of Q fever. Journal of Milk & Food Technology 24 351355Google Scholar
Enright, JB, Sadler, WW & Thomas, RC 1957 Thermal inactivation of Coxiella burnetii and its relation to pasteurization of milk Public Health Monograph No. 47, PHS Pub. No. 517 U.S. Government Printing Office., Washington, DCGoogle Scholar
Gale, P, Kelly, L, Mearns, R, Duggan, J & Snary, EL 2015 Q fever through consumption of unpasteurized milk and milk products – a risk profile and exposure assessment. Journal of Applied Microbiology 118 10831095Google Scholar
Howe, GB, Loveless, BM, Norwood, D, Craw, P, Waag, D, England, M, Lowe, JR, Courtney, BC, Pitt, ML & Kulesh, DA 2009 Real-time PCR for the early detection and quantification of Coxiella burnetii as an alternative to the murine bioassay. Molecular & Cellular Probes 23 127131Google Scholar
Kersh, GJ, Priestley, RA, Hornstra, HM, Self, JS, Fitzpatrick, KA, Biggerstaff, BJ, Keinm, P, Pearson, T & Massung, RF 2016 Genotyping and axenic growth of Coxiella burnetii isolates found in the United States environment. Vector Borne Zoonotic Diseases 16 588594Google Scholar
Lovett, J, Wesley, IV, Vandermaaten, MJ, Bradshaw, JG, Francis, DW, Crawford, RG, Donnelly, CW & Messer, JW 1990 High-temperature short-time pasteurization inactivated Listeria monocytogenes. Journal of Food Protection 53 734738Google Scholar
Omsland, A, Beare, PA, Hill, J, Cockrell, D, Howe, D, Hansen, B, Samuel, JE & Heinzen, RA 2011 Isolation from animal tissue and genetic transformation of Coxiella burnetii are facilitated by an improved axenic growth medium. Applied & Environmental Microbiology 77 32703725Google Scholar
Pearce, LE, Smythe, BW, Crawford, RA, Oakley, E, Hathaway, SC & Shephard, JM 2012 Pasteurization of milk: The heat inactivation kinetics of milk-borne dairy pathogens under commercial-type conditions of turbulent flow. Journal of Dairy Science 95 2035Google Scholar
Stewart, D, Shieh, YC, Tortorello, M, Kukreja, A, Shazer, A & Schlesser, J 2015 Quantitation of viable Coxiella burnetii in milk using an integrated cell culture-polymerase chain reaction (ICC-PCR) assay. Journal of Dairy Research 82 478484Google Scholar
Supplementary material: PDF

Shi et al. supplementary material

Shi et al. supplementary material 1

Download Shi et al. supplementary material(PDF)
PDF 428.6 KB