title: Microbiological surveillance of a bovine raw milk farm through multiplex real-time PCR.
abstract: Raw milk is increasingly appreciated by consumers but can be contaminated by a variety of zoonotic pathogens. Therefore, preventive measures, such as on-farm hazard analysis critical control point (HACCP) programs, must be applied to protect consumers. The aim of the present study was the comparison of a multiplex real-time polymerase chain reaction (PCR) assay with a culture-based approach in an on-farm quality assurance program for the detection of Escherichia coli O157, Salmonella spp., and Listeria monocytogenes in bulk tank milk, in-line milk filters, manure, and feces. Results revealed that the real-time PCR was more sensitive in detecting E. coli O157 than the culture method in filters (48% vs. 4% positive), manure (93% vs. 7% positive) and feces (60% vs. 4% positive). The two methods were equally efficient in detecting L. monocytogenes (8% of filters), while Salmonella spp. was not detected in any sample. In conclusion, the real-time PCR, by reducing analysis time to two working days, can be proposed as a useful tool in the raw milk primary production setting as a rapid and user-friendly screening method.
authors.affiliation: Dipartimento di Scienze Biomolecolari, Università degli Studi di Urbino "Carlo Bo," Urbino, Italy. giulia.amagliani@uniurb.it
authors.forename: Giulia
authors.initials: G
authors.lastname: Amagliani
authors.forename: Annalisa
authors.initials: A
authors.lastname: Petruzzelli
authors.forename: Enrica
authors.initials: E
authors.lastname: Omiccioli
authors.forename: Franco
authors.initials: F
authors.lastname: Tonucci
authors.forename: Mauro
authors.initials: M
authors.lastname: Magnani
authors.forename: Giorgio
authors.initials: G
authors.lastname: Brandi
mesh_terms: D000818:Animals; D002417:Cattle; D003612:Dairying / Q000295:instrumentation; D004864:Equipment and Supplies / Q000382:microbiology; D019453:Escherichia coli O157 / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D005243:Feces / Q000382:microbiology; D005260:Female; D005511:Food Handling; D005513:Food Inspection / Q000379:methods*; D005517:Foodborne Diseases / Q000517:prevention & control; D007218:Industrial Microbiology / Q000379:methods*; D007558:Italy; D008089:Listeria monocytogenes / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D008297:Male; D008372:Manure / Q000382:microbiology; D008892:Milk / Q000382:microbiology*; D058889:Molecular Typing / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction; D011786:Quality Control; D060888:Real-Time Polymerase Chain Reaction; D012475:Salmonella / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D013997:Time Factors
chemical_list: D008372:Manure