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Forming Bacteria in Foods Ανάπτυξη Και Εφαρμογή Μαθηματικών Μοντέλων Πρόβλεψης Της Συμπεριφοράς Θερμόφιλων Σποριογόνων Βακτηρίων Στα Τρόφιμα

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Forming Bacteria in Foods Ανάπτυξη Και Εφαρμογή Μαθηματικών Μοντέλων Πρόβλεψης Της Συμπεριφοράς Θερμόφιλων Σποριογόνων Βακτηρίων Στα Τρόφιμα ARISTOTLE UNIVERSITY OF THESSALONIKI FACULTY OF AGRICULTURE, FORESTRY AND NATURAL ENVIRONMENT SCHOOL OF AGRICULTURE ΑΡΙΣΤΟΤΕΛΕΙΟ ΠΑΝΕΠΙΣΤΗΜΙΟ ΘΕΣΣΑΛΟΝΙΚΗΣ ΣΧΟΛΗ ΓΕΩΠΟΝΙΑΣ, ΔΑΣΟΛΟΓΙΑΣ ΚΑΙ ΦΥΣΙΚΟΥ ΠΕΡΙΒΑΛΛΟΝΤΟΣ ΤΜΗΜΑ ΓΕΩΠΟΝΙΑΣ MYRSINI N. KAKAGIANNI Biologist, MSc ΜΥΡΣΙΝΗ Ν. ΚΑΚΑΓΙΑΝΝΗ Πτυχιούχος Βιολόγος, MSc Development and application of predictive models for the growth of thermophilic spore - forming bacteria in foods Ανάπτυξη και εφαρμογή μαθηματικών μοντέλων πρόβλεψης της συμπεριφοράς θερμόφιλων σποριογόνων βακτηρίων στα τρόφιμα PhD THESIS ΔΙΔΑΚΤΟΡΙΚΗ ΔΙΑΤΡΙΒΗ THESSALONIKI / 2018 ΘΕΣΣΑΛΟΝΙΚΗ / 2018 MYRSINI N. KAKAGIANNI Biologist, MSc Development and application of predictive models for the growth of thermophilic spore - forming bacteria in foods Ph.D. THESIS Submitted at School of Agriculture of Aristotle University of Thessaloniki DATE OF ORAL PRESENTATION: Friday 13 July 2018 THESIS COMMITTEE (Decision ΓΣΕΣ 122/01-06-2018): FULL NAME LEVEL ROLE INSTITUTION Konstantinos P. Aristotle University of Professor Supervisor Koutsoumanis Thessaloniki Member of Three – Aristotle University of Konstantinos Biliaderis Professor member Committee Thessaloniki Assistant Member of Three – Aristotle University of Eugenios Katsanidis Professor member Committee Thessaloniki Agricultural University of George – John Nychas Professor Member Athens Associate Aristotle University of Apostolos S. Angelidis Member Professor Thessaloniki Associate Agricultural University of Efstathios Panagou Member Professor Athens Assistant Aristotle University of Athanasia Goula Member Professor Thessaloniki ΜΥΡΣΙΝΗ Ν. ΚΑΚΑΓΙΑΝΝΗ Πτυχιούχος Βιολόγος, MSc Ανάπτυξη και εφαρμογή μαθηματικών μοντέλων πρόβλεψης της συμπεριφοράς θερμόφιλων σποριογόνων βακτηρίων στα τρόφιμα ΔΙΔΑΚΤΟΡΙΚΗ ΔΙΑΤΡΙΒΗ Υποβλήθηκε στο Τμήμα Γεωπονίας του Αριστοτελείου Πανεπιστημίου Θεσσαλονίκης ΗΜΕΡΟΜΗΝΙΑ ΥΠΟΣΤΗΡΙΞΗΣ: Παρασκευή 13 Ιουλίου 2018 ΕΞΕΤΑΣΤΙΚΗ ΕΠΙΤΡΟΠΗ (Απόφαση ΓΣΕΣ 122/01-06-2018): ΟΝΟΜΑΤΕΠΩΝΥΜΟ ΒΑΘΜΙΔΑ ΙΔΙΟΤΗΤΑ ΙΔΡΥΜΑ Κωνσταντίνος Π. Αριστοτέλειο Πανεπιστήμιο Καθηγητής Επιβλέπων Κουτσουμανής Θεσσαλονίκης Μέλος Κωνσταντίνος Αριστοτέλειο Πανεπιστήμιο Καθηγητής Τριμελούς Μπιλιαδέρης Θεσσαλονίκης Επιτροπής Μέλος Επίκουρος Αριστοτέλειο Πανεπιστήμιο Ευγένιος Κατσανίδης Τριμελούς Καθηγητής Θεσσαλονίκης Επιτροπής Γεώργιος – Ιωάννης Γεωπονικό Πανεπιστήμιο Καθηγητής Μέλος Νυχάς Αθηνών Αναπληρωτής Αριστοτέλειο Πανεπιστήμιο Απόστολος Σ. Αγγελίδης Μέλος Καθηγητής Θεσσαλονίκης Αναπληρωτής Γεωπονικό Πανεπιστήμιο Ευστάθιος Πανάγου Μέλος Καθηγητής Αθηνών Επίκουρος Αριστοτέλειο Πανεπιστήμιο Αθανασία Γούλα Μέλος Καθηγήτρια Θεσσαλονίκης © Myrsini N. Kakagianni, 2018 © PhD thesis, Aristotle University of Thessaloniki, Thessaloniki, Greece, 2018 Development and application of predictive models for the growth of thermophilic spore - forming bacteria in foods Ανάπτυξη και εφαρμογή μαθηματικών μοντέλων πρόβλεψης της συμπεριφοράς θερμόφιλων σποριογόνων βακτηρίων στα τρόφιμα ISBN «Η έγκριση της παρούσης διδακτορικής διατριβής από το Τμήμα Γεωπονίας δεν υποδηλώνει αποδοχή των γνωμών και απόψεων του συγγραφέως» (Ν. 5343/1932, άρθρο 202, παρ. 2) Μέρος της παρούσας διατριβής υλοποιήθηκε στο πλαίσιο της Δράσης «Συνεργασία» ΕΣΠΑ 2007 - 2013 και του προγράμματος Erasmus+ και συγχρηματοδοτήθηκε από: -Ευρωπαϊκή ΄Ενωση - Ευρωπαϊκό Κοινωνικό Ταμείο -Γενική Γραμματεία Έρευνας και Τεχνολογίας στον Αποστόλη και το Νικόλα Ευχαριστίες Αρχικά, θα ήθελα να εκφράσω τη βαθύτατη ευγνωμοσύνη μου στον κ. Κωνσταντίνο Κουτσουμανή για την πολύτιμη καθοδήγηση και τη συνεχή στήριξή του κατά τη διάρκεια όλων αυτών των ετών. Τον ευχαριστώ θερμά για το χρόνο, τις ιδέες και την εμπιστοσύνη που έδειξε στο πρόσωπό μου δίνοντάς μου την ευκαιρία να συνεχίσω τις πανεπιστημιακές μου σπουδές με την εκπόνηση της διδακτορικής αυτής μελέτης και να εξελιχθώ ως ερευνήτρια. Τον ευχαριστώ, επίσης, που μου έμαθε τη σημασία της συνεργασίας των επιστημών αλλά και τη μετάδοση της γνώσης από γενιά σε γενιά προκειμένου να είμαστε σε θέση να προβούμε σε αξιόλογα αποτελέσματα. Θεωρώ τον εαυτό μου τυχερό που ήμουν ένα από τα μέλη της ερευνητικής του ομάδας καθώς μου μετέδωσε πολλές και πλούσιες γνώσεις στο χώρο της Ποσοτικής Μικροβιολογίας. Επίσης, θα ήθελα να ευχαριστήσω όλα τα μέλη της εξεταστικής μου επιτροπής, τους καθηγητές κ. Κωνσταντίνο Μπιλιαδέρη, κ. Ευγένιο Κατσανίδη, κ. Γεώργιο – Ιωάννη Νυχά, κ. Ευστάθιο Πανάγου, κ. Απόστολο Αγγελίδη και κα. Αθανασία Γούλα, που αξιολόγησαν αυτή τη διατριβή και ενήργησαν ως «αντίπαλοι» κατά την υπεράσπισή της. Θα ήθελα να εκφράσω τις θερμές μου ευχαριστίες στο Βασίλη Βαλδραμίδη, Αν. Καθηγητή του Πανεπιστημίου της Μάλτας, για την εμπιστοσύνη του και την ευκαιρία που μου έδωσε να υλοποιήσω ένα μικρό μέρος της διδακτορικής αυτής μελέτης, υπό την επίβλεψή του, στο εργαστήριό του. Εκτιμώ βαθιά την εξαίρετη συνεργασία μας, την πολύτιμη καθοδήγησή του και την ανιδιοτελή μετάδοση των γνώσεών του στο αντικείμενο της ποσοτικής μικροβιολογίας. Ιδιαίτερες ευχαριστίες θα ήθελα να εκφράσω στο δάσκαλό μου κ. Μηνά Γιάγκου για την εμπιστοσύνη που έδειξε στο πρόσωπό μου και την ευκαιρία που μου έδωσε να γνωρίσω τον κόσμο της Ανοσοβιολογίας. Έπαιξε καθοριστικό ρόλο στη ζωή μου καθώς ήταν ο άνθρωπος που μετά από συζητήσεις με παρότρυνε να συνεχίσω τις μεταπτυχιακές μου σπουδές εξερευνώντας το χώρο των τροφίμων. Ένα μεγάλο ευχαριστώ οφείλω στους συναδέλφους, συνεργάτες και συμφοιτητές, συμπεριλαμβανομένων και των προπτυχιακών φοιτητών (Φωτεινή Μπόμπολα, Φανή Τσίνα, Αγγελική Χωραΐτη, Χριστίνα Νταμπούδη, Όλια Χαρισμιάδου, Μαρία Ρέπα, Μαίρη Τσιλφίδου και Μαρία Παπακρασά), για τη βοήθεια και ουσιαστική συμβολή τους καθ’ όλη τη διάρκεια της διδακτορικής διατριβής. Την ιδιαίτερη ευγνωμοσύνη και αγάπη μου θα ήθελα να εκφράσω στη Μαίρη Γουγουλή, για την άριστη συνεργασία και την αμέριστη βοήθεια οποτεδήποτε της ζήτησα, και τη Δάφνη Δημακοπούλου – Παπάζογλου, για τη σημαντική συμβολή της σε παρατηρήσεις και διορθώσεις της παρούσας διδακτορικής διατριβής, τη φιλία και συμπαράστασή της όλα αυτά τα χρόνια, ενώ παράλληλα μαζί με την Αλεξία Λιανού, Ζάφη Ασπρίδου, και Ζαχαρένια Ζαμπουλάκη ζήσαμε αξέχαστες στιγμές, όμορφες και δύσκολες, δημιουργώντας ένα ιδανικό κλίμα συνεργασίας, δημιουργικότητας και σκληρής δουλειάς. Είστε πλέον αναπόσπαστο κομμάτι της καρδιάς μου. Ιδιαιτέρως θα ήθελα να ευχαριστήσω τους φίλους μου Δημήτρη Βασιλείου για την καθοριστική βοήθειά του όσον αφορά τη συλλογή των θερμοκρασιακών δεδομένων καθώς και την Ελένη Νάστα για την επιμέλεια του εξωφύλλου της συγκεκριμένης διατριβής. Τέλος, ένα μεγάλο ευχαριστώ θα ήθελα να εκφράσω στον Αποστόλη μου για τις διορθώσεις του στην παρούσα διδακτορική διατριβή, την αγάπη, την υποστήριξη και την υπομονή του όλα αυτά τα χρόνια, ενώ πάρα πολλές φορές με βοήθησε να επαναπροσδιορίσω τις προτεραιότητές μου. Είμαι παντοτινά ευγνώμων στους γονείς μου, Μαίρη και Νίκο, αλλά και την αδερφή μου, Βίκυ, για την αγάπη τους, τη συνεχή και ανεξάντλητη συμπαράσταση και υποστήριξη σε κάθε μου απόφαση όλα τα χρόνια των σπουδών μου. Μυρσίνη Contents List of Tables 13 List of Figures 15 List of Abbreviations 19 Abstract 25 Περίληψη 29 Chapter 1 Literature Review and Thesis Outline 33 Chapter 2 Development and application of Geobacillus 97 stearothermophilus growth model for predicting spoilage of evaporated milk Chapter 3 Development and validation of predictive models for the 117 effect of storage temperature and pH on the growth boundaries and kinetics of Alicyclobacillus acidoterrestris ATCC 49025 in fruit drinks Chapter 4 Effect of storage temperature on the lag time of Geobacillus 141 stearothermophilus individual spores Chapter 5 Mapping the risk of evaporated milk spoilage in the 159 Mediterranean region based on the effect of temperature conditions on Geobacillus stearothermophilus growth Chapter 6 General Discussion and potential applications of the 174 developed models to the food industry References 186 List of Publications 233 List of Tables Table 2.1. Estimated values and statistics for the parameters of the cardinal 106 model with inflection (Equation (2.1)) describing the effect of temperature on the maximum specific growth rate (μmax) of Geobacillus stearothermophilus ATCC 7953 in tryptone soy broth. Table 2.2. Comparison between observed and predicted spoilage time of the 115 evaporated milk, stored under nonisothermal conditions, by Geobacillus stearothermophilus ATCC 7953. Table 3.1. Estimated values and fitting statistics for the parameters of the 129 logistic polynomial regression model for the combined temperature and pH limits of Alicyclobacillus acidoterrestris growth in K broth. Table 3.2. Probability of Alicyclobacillus acidoterrestris growth predicted by the 129 growth/no growth boundaries model for different combinations of temperature and pH. Table 3.3. Effect of pH on the maximum specific growth rate (μmax) of 131 Alicyclobacillus acidoterrestris in K broth at 48 °C. Points represent the observed μmax values, the solid line corresponds to the fitting on the cardinal pH model to the data, and the dotted lines depict the 95% confidence and prediction intervals. Table 3.4. Comparison between observed behavior of Alicyclobacillus 133 acidoterrestris and probability of growth predicted by the developed growth/no growth interface model in commercial pasteurized fruit drinks tested in validation studies Table 4.1. Statistics of the individual spore lag time (λ) of Geobacillus 149 stearothermophilus. Table 4.2. Parameter estimation of the Gamma distribution fitted to the 149 individual spore lag times of Geobacillus stearothermophilus. The probability density function of the Gamma distribution
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