title: Estimation of Mycobacterium avium subsp. paratuberculosis load in raw bulk tank milk in Emilia-Romagna Region (Italy) by qPCR.
abstract: Consumption of milk and dairy products is considered one of the main routes of human exposure to Mycobacterium avium subsp. paratuberculosis (MAP). Quantitative data on MAP load in raw cows' milk are essential starting point for exposure assessment. Our study provides this information on a regional scale, estimating the load of MAP in bulk tank milk (BTM) produced in Emilia-Romagna region (Italy). The survey was carried out on 2934 BTM samples (88.6% of the farms herein present) using two different target sequences for qPCR (f57 and IS900). Data about the performances of both qPCRs are also reported, highlighting the superior sensitivity of IS900-qPCR. Seven hundred and eighty-nine samples tested MAP-positive (apparent prevalence 26.9%) by IS900 qPCR. However, only 90 of these samples were quantifiable by qPCR. The quantifiable samples contained a median load of 32.4 MAP cells mL(-1) (and maximum load of 1424 MAP cells mL(-1) ). This study has shown that a small proportion (3.1%) of BTM samples from Emilia-Romagna region contained MAP in excess of the limit of detection (1.5 × 10(1) MAP cells mL(-1) ), indicating low potential exposure for consumers if the milk subsequently undergoes pasteurization or if it is destined to typical hard cheese production.
authors.affiliation: Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna, National Reference Centre for Paratuberculosis, Strada Faggiola 1, loc., Gariga - Podenzano (PC), 29027, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Ricchi
authors.affiliation: Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna, National Reference Centre for Paratuberculosis, Strada Faggiola 1, loc., Gariga - Podenzano (PC), 29027, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Savi
authors.affiliation: Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna, Centro di Referenza Nazionale per i Rischi Emergenti in Sicurezza Alimentare, 20133, Milan, Italy.|Risk Analysis Unit, Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna, Via dei Mercati 13/A, Parma, 43121, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Bolzoni
authors.affiliation: Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna, Centro di Referenza Nazionale per i Rischi Emergenti in Sicurezza Alimentare, 20133, Milan, Italy.|Risk Analysis Unit, Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna, Via dei Mercati 13/A, Parma, 43121, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Pongolini
authors.affiliation: Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, 97 Lisburn Road, Belfast, BT9 7BL, Northern Ireland, United Kingdom.
authors.forename: Irene R
authors.initials: IR
authors.lastname: Grant
authors.affiliation: Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna, National Reference Centre for Paratuberculosis, Strada Faggiola 1, loc., Gariga - Podenzano (PC), 29027, Italy.
authors.forename: Caterina
authors.initials: C
authors.lastname: De Cicco
authors.affiliation: Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna, National Reference Centre for Paratuberculosis, Strada Faggiola 1, loc., Gariga - Podenzano (PC), 29027, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Cerutti
authors.affiliation: Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna, National Reference Centre for Paratuberculosis, Strada Faggiola 1, loc., Gariga - Podenzano (PC), 29027, Italy.
authors.forename: Giuliana
authors.initials: G
authors.lastname: Cammi
authors.affiliation: Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna, National Reference Centre for Paratuberculosis, Strada Faggiola 1, loc., Gariga - Podenzano (PC), 29027, Italy.
authors.forename: Chiara A
authors.initials: CA
authors.lastname: Garbarino
authors.affiliation: Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna, National Reference Centre for Paratuberculosis, Strada Faggiola 1, loc., Gariga - Podenzano (PC), 29027, Italy.
authors.forename: Norma
authors.initials: N
authors.lastname: Arrigoni
mesh_terms: D000818:Animals; D058491:Bacterial Load / Q000379:methods / Q000662:veterinary*; D002417:Cattle; D004269:DNA, Bacterial / Q000235:genetics; D006801:Humans; D007558:Italy; D057230:Limit of Detection; D008892:Milk / Q000382:microbiology*; D016927:Mycobacterium avium subsp. paratuberculosis / Q000302:isolation & purification*; D010283:Paratuberculosis / Q000175:diagnosis* / Q000382:microbiology; D061352:Pasteurization; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D004269:DNA, Bacterial
keywords: Bulk tank milk; Mycobacterium avium subsp. paratuberculosis; peptide magnetic separation; qPCR; quantification