Σκοπός: Ο προσδιορισμός των επιπέδων συγκέντρωσης καταλοίπων κτηνιατρικών παρασιτοκτόνων ουσιών (ανθελμινθικών), σε νωπό γάλα μηρυκαστικών από περιοχές της Νοτίου Ελλάδος και η εκτίμηση της πρόσληψής τους από τον άνθρωπο. Συγκεκριμένα εξετάσθηκαν οι ουσίες, Albendazole, Febantel, Fenbendazole, Mebendazole και κάποιοι μεταβολίτες τους (Albendazole sulfoxide, Albendazole sulfone, Fenbendazole sulfone) στο νωπό γάλα προβάτων, αιγών και βοοειδών. Μέθοδοι: Χημική ανάλυση δειγμάτων γάλακτος, με τη βοήθεια Υγρής Χρωματογραφίας Υψηλής Απόδοσης και ανιχνευτή Συστοιχίας Διόδων, Υπεριώδους (UV). Χρήση βάσης δεδομένων σχετικά με την εκτίμηση της ημερήσιας κατανάλωσης γάλακτος και Φέτας (λήψη προσωπικών συνεντεύξεων, συμπλήρωση ερωτηματολογίων, συχνοτήτων κατανάλωσης τροφίμων, από δείγμα 723 μαθητών ηλικίας 10-12 ετών, από τη Νότια Ελλάδα). Αποτελέσματα: 34 από τα 123 δείγματα γάλακτος βρέθηκαν να περιέχουν κατάλοιπα των διερευνώμενων ουσιών, εκ των οποίων τα 11 υπερέβαιναν τα θεσπισμένα από την Ε.Ε. ανώτατα επιτρεπτά όρια καταλοίπων. Η Εκτιμώμενη Ημερήσια Πρόσληψη των ουσιών αυτών, από το εξετασθέν δείγμα πληθυσμού, κυμαίνεται από 0,4-15,9% της Αποδεκτής Ημερήσιας Πρόσληψης, ανάλογα με την ουσία και το τρόφιμο (νωπό αγελαδινό γάλα ή τυρί Φέτα). Υπάρχει γεωγραφική διακύμανση στη συχνότητα εμφάνισης των καταλοίπων η οποία είναι ιδιαίτερα αυξημένη σε περιοχές οι οποίες στερούνται επαρκών υπηρεσιών ελέγχου τροφίμων. Συμπεράσματα: Από την παρούσα εργασία προκύπτει ότι 11.4% των δειγμάτων γάλακτος, περιέχει κατάλοιπα ανθελμινθικών ουσιών, σε επίπεδα συγκεντρώσεων που υπερβαίνουν το ανώτατο επιτρεπτό όριο (έως και 7 φορές για τη Febantel). Το εύρημα αυτό, εγείρει ερωτήματα για την πλήρη εφαρμογή της Ο.Κ.Π.. Ωστόσο η κατανάλωση νωπού γάλακτος το οποίο περιέχει κατάλοιπα των υπό διερεύνηση ενώσεων, στα προαναφερθέντα επίπεδα συγκεντρώσεων, δεν οδηγεί σε υπέρβαση της Ανώτερης Ημερήσιας Πρόσληψης. Από την άλλη πλευρά, είναι απαραίτητο να τονιστεί ότι οι ουσίες αυτές είναι δυνατό να ανιχνευθούν και σε άλλα τρόφιμα ζωικής προέλευσης για τα οποία δεν έχουν θεσπιστεί MRLs και συνεπώς δε διεξάγονται οι αντίστοιχοι έλεγχοι. Κατά συνέπεια, είναι απαραίτητη η εκπαίδευση των κτηνοτρόφων σε θέματα Ο.Κ.Π., η εντατικοποίηση των ελέγχων και η στελέχωση των κρατικών υπηρεσιών, έτσι ώστε να αντισταθμίζονται οι ελλείψεις των νόμων και να αποφευγονται πιθανές δυσμενείς συνέπειες για τη δημόσια υγεία. / Aim: The determination of the concentration levels, of veterinary parasiticide drug (anthelmintics) residues, in ruminants’ raw milk, from regions of Southern Greece, and the residues’ intake estimation for humans. In particular, the investigated, residues were Albendazole, Febantel, Fenbendazole, Mebendazole and some of their metabolites (Albendazole sulfoxide, Albendazole sulfone, Fenbendazole sulfone), in sheep, goat and bovine raw milk. Methods: Chemical analysis of raw milk samples with High Performance Liquid Chromatography and UV Diode Array Detector. A databank concerning the daily consumption of milk and feta cheese was utilized (personal interviews and filling in of food frequency questionnaires, from a population sample of 723 pupils aged from 10-12 years old, in Southern Greece). Results: 34 out of the 123 milk samples, were found to contain residues of the investigated compounds and 11 of the contaminated samples, contained residues, exceeding the EU established MRLs. The Estimated Daily Intake for these residues resulting from the interviewed population sample, was ranging from 0,4 to 15,9% of the Acceptable Daily Intake, depending on the substance and the kind of food (raw bovine milk or feta cheese). There is a geographical variation concerning the residue occurrence, which is high in regions where food control agencies are poorly staffed. Conclusions: The present study indicates that 11.4% of milk samples analysed, contains concentration levels of anthelmintic residues above the maximum residue limit (up to 7 times for Febantel). This finding raises questions about the full implementation of Good Veterinary Practice. However, consumption of raw milk containing the aforementioned concentration levels of the compounds under investigation, does not result to exceedance of the Acceptable Daily Intake. On the other hand, it is necessary to emphasize that these substances are possible to be detected in other foods of animal origin for which no MRLs have been established and therefore the respective controls are not carried out. Consequently, training of stock-farmers on GVP, intensification of controls and staffing of government agencies, are needed to counterbalance deficiencies of laws and avoid potential adverse effects on public health.

The novel hypothesis that antimicrobial growth promoters (AGP) function by modulating the mammalian immune system was tested. Sampling methods to characterize the mucosa-associated microbiota of the murine intestine by terminal restriction fragment length polymorphism (T-RFLP) analysis indicated that direct plug extraction was superior to wash methods. Using T-RFLP analysis, non-therapeutic administration of chlortetracycline (CTC) and sulfamethazine to beef cattle did not affect the composition of bacterial communities associated with intestinal mucosa and in digesta, with exception of those associated with mucosa of the proximal jejunum. Similarly, oral administration of non-therapeutic concentrations of CTC did not affect the mucosa-associated microbiota of the murine intestine. Oral administration of nontherapeutic concentrations of CTC prevented weight loss, reduced pathologic changes, modulated transcription levels of inflammatory cytokines in C. rodentium-infected mice, and did not consistently affect the colonic microbiota. These findings support the hypothesis that AGP primarily function by modulating the intestinal immune system. / xiv, 160 leaves ; 29 cm

Veterinary pharmaceuticals are routinely used in livestock production to treat diseases, prevent infections, and promote growth. However, the potential release of pharmaceuticals from agricultural activities has raised concerns because they may pose detrimental effects to the ecosystems and human health, for example fostering the evolution of antibiotic-resistant bacteria in the natural environment. A better understanding of the environmental fate of veterinary pharmaceuticals is critical to properly assess and mitigate their risks. This dissertation focuses on a major group of veterinary pharmaceuticals, ionophore antibiotics (IPAs), which is sold at over 4 million kilograms per year and constitutes more than one third of the total antibiotic consumption by the livestock industry in the U.S. Despite the extensive usage of IPAs, their environmental fate was not well-understood. Therefore, this study aimed at achieving a comprehensive understanding of the occurrence, persistence, and transformation of IPAs from poultry litter before and after applications to the agricultural lands. Three of the most commonly used members of IPAs were investigated in this study: monensin (MON), salinomycin (SAL), and narasin (NAR). Based on the common management practices of poultry litter, the potential abiotic and biotic transformation reactions of IPAs were examined under varying conditions relevant to the water-soil-litter systems. This dissertation consists of three sections. First, a robust analytical method was developed to quantify IPAs in various environmental compartments, especially in high organic-containing matrices such as poultry litter, and soil and runoff from litter-fertilized lands. Efforts were made to optimize the analytical method with respect to improving extraction recovery, reducing matrix effects, and validating a surrogate standard. Second, lab-scale experiments were set up to determine the chemical properties of IPAs in aqueous environments and to study the abiotic transformation of IPAs, including hydrolysis and photolysis. The results showed that IPAs are prone to hydrolytic transformation in acidic environments, which are likely to be encountered in acidic soils, alum-amended litter (alum: Al₂(SO₄)₃•12H₂O), and acidic runoff. Multiple transformation pathways were proposed based on the identified hydrolysis products. It is also noteworthy that the hydrolysis products of MON still exhibited a toxic effect on the selected microorganism (Bacillus subtilis). SAL and NAR were found to undergo direct photolysis under both UV light and sunlight irradiation. In natural water matrix, IPAs were also degraded by indirect photolysis with hydroxyl radicals generated by light-excited nitrate. Dissolved organic matter can shield IPAs from light and slow down their photolysis. Third, the biodegradation potential of IPAs was first tested in litter and soil microcosms. Factor analysis was conducted to delineate the interaction of water and temperature on IPA degradation in the litter. Litter-fertilized and non-fertilized soil microcosms were compared on the degradation of MON and SAL. Furthermore, the inhibition and biotransformation potential of IPAs were assessed under different redox conditions with litter-enriched cultures. Inhibition tests focused on examining IPAs’ impact on microbial community functions, including denitrification, sulfate-reduction, and methane production. Biodegradation tests were conducted with different electron acceptors, including oxygen, nitrate, sulfate, and organic carbons, with efforts to elucidate primary biotransformation products. On the basis of the results obtained in this study, several recommendations on litter management and IPA selection were made to help mitigate the release and transport of IPAs, as well as enhance their degradation. Overall, this study significantly improved the understanding of the environmental fate of IPAs and the obtained knowledge can aid proper selection of IPAs and management strategies in future applications to minimize the risks of antibiotic micropollutants in the environment.

The use of veterinary drugs continues to be a challenge since some farmers use drugs for medicinal purposes, while others use them as growth promoters without observing the set withdrawal times. In emerging economies, challenges are experienced in residue testing to ensure residue free animal products. It is therefore necessary to develop a simple, environmentally friendly and cost effective extraction method for veterinary drugs residues. A dispersive liquid-liquid micro-extraction method for simultaneous determination of chloramphenicol, florfenicol and thiamphenicol residues in water, bovine muscle and milk was developed optimised and validated. Analysis was performed with liquid chromatography coupled to electron spray ionization tandem mass spectrometry in negative ion multiple reaction monitoring mode. Dispersive liquid-liquid micro-extraction method development involved optimisation of parameters such as type and volume of extraction solvent, type and volume of disperser solvent and pH. The optimum extraction solvent was dichloromethane at a volume of 250 μL while the optimum disperser solvent was acetonitrile (1 mL) Extraction was found to be optimum at pH 7. The developed method was validated according to Commission decision 2002/657/EC. Linearity. Linearity was observed for water, bovine milk and muscle in the ranges of 0.3 μg kg-1 to 0.9 μg kg-1, 25 μg kg-1 to 100 μg kg-1 and 50 μg kg-1 to 200 μg kg-1 for chloramphenicol, thiamphenicol and florfenicol respectively with regression coefficients ranging between 0.9941 and 0.9999. Limit of detections for CAP, FFC & THAP were 0.082, 3.31, and 2.21 μg kg-1 respectively while Limit of quantifications were 0.163, 7.51 and 6.84 μg kg-1 respectively. The recoveries for water, bovine milk and bovine muscle ranged between 87.2% and 102%. In this study the validated dispersive liquid-liquid micro-extraction has been found to perform very well not only in the water matrix but also in the more complex matrices like bovine milk and muscle. / Chemistry / M. Sc. (Chemistry)