Trattamenti ad aria calda per la decontaminazione superficiale delle uova in guscio

In order to improve the animal welfare, the Council Directive 1999/74/EC (defining minimum standards for the welfare of laying hens) will ban conventional cage systems since 2012, in favour of enriched cages or floor systems. As a consequence an increased risk of bacterial contamination of eggshell is expected (EFSA, 2005). Furthermore egg-associated salmonellosis is an important public health problem throughout the world (Roberts et al., 1994). In this regard the introduction of efficient measures to reduce eggshell contamination by S. Enteritidis or other bacterial pathogens, and thus to prevent any potential or additional food safety risk for Human health, may be envisaged. The hot air pasteurization can be a viable alternative for the decontamination of the surface of the egg shell. Few studies have been performed on the decontamination power of this technique on table eggs (Hou et al, 1996; James et al., 2002). The aim of this study was to develop innovative techniques to remove surface contamination of shell eggs by hot air under natural or forced convection. Initially two simplified finite element models describing the thermal interaction between the air and egg were developed, respectively for the natural and forced convection. The numerical models were validated using an egg simulant equipped by type-K thermocouple (Chromel/Alumel). Once validated, the models allowed the selection of a thermal cycle with an inner temperature always lower than 55°C. Subsequently a specific apparatus composed by two hot air generators, one cold air generator and rolling cylinder support, was built to physically condition the eggs. The decontamination power of the thermal treatments was evaluated on shell eggs experimentally inoculated with either Salmonella Enteritidis, Escherichia coli, Listeria monocytogenes and on shell eggs containing only the indigenous microflora. The applicability of treatments was further evaluated by comparing quality traits of treated and not treated eggs immediately after the treatment and after 28 days of storage at 20°C. The results showed that the treatment characterized by two shots of hot air at 350°C for 8 sec, spaced by a cooling interval of 32 (forced convection), reduce the bacterial population of more than 90% (Salmonella enteritidis and Listeria monocytogenes). No statistically significant results were obtained comparing E. coli treated and not treated eggs as well as indigenous microflora treated and not treated eggs. A reduction of 2.6 log was observed on Salmonella enteritidis load of eggs immediately after the treatment in oven at 200°C for 200 minutes (natural convection). Furthermore no detrimental effects on quality traits of treated eggs were recorded. These results support the hot air techniques for the surface decontamination of table eggs as an effective industrial process.

Atmospheric plasma processes for microbial inactivation: food applications and stress response in Listeria monocytogenes

This PhD thesis is focused on cold atmospheric plasma treatments (GP) for microbial inactivation in food applications. In fact GP represents a promising emerging technology alternative to the traditional methods for the decontamination of foods. The objectives of this work were to evaluate: - the effects of GP treatments on microbial inactivation in model systems and in real foods; - the stress response in L. monocytogenes following exposure to different GP treatments. As far as the first aspect, inactivation curves were obtained for some target pathogens, i.e. Listeria monocytogenes and Escherichia coli, by exposing microbial cells to GP generated with two different DBD equipments and processing conditions (exposure time, material of the electrodes). Concerning food applications, the effects of different GP treatments on the inactivation of natural microflora and Listeria monocytogenes, Salmonella Enteritidis and Escherichia coli on the surface of Fuji apples, soya sprouts and black pepper were evaluated. In particular the efficacy of the exposure to gas plasma was assessed immediately after treatments and during storage. Moreover, also possible changes in quality parameters such as colour, pH, Aw, moisture content, oxidation, polyphenol-oxidase activity, antioxidant activity were investigated. Since the lack of knowledge of cell targets of GP may limit its application, the possible mechanism of action of GP was studied against 2 strains of Listeria monocytogenes by evaluating modifications in the fatty acids of the cytoplasmic membrane (through GC/MS analysis) and metabolites detected by SPME-GC/MS and 1H-NMR analyses. Moreover, changes induced by different treatments on the expression of selected genes related to general stress response, virulence or to the metabolism were detected with Reverse Transcription-qPCR. In collaboration with the Scripps Research Institute (La Jolla, CA, USA) also proteomic profiles following gas plasma exposure were analysed through Multidimensional Protein Identification Technology (MudPIT) to evaluate possible changes in metabolic processes.

Role of bioactive compounds in the gastrointestinal host-pathogen interaction of poultry and swine

The better understanding of mechanisms at the basis of host-pathogen interaction can represent a valid tool to increase productivity and contain economic losses in animal production through the maintenance of intestinal homeostasis. With this project, three preliminary in vitro studies were conducted with the aim of investigating how bioactive compounds could influence mechanisms of host-pathogen interaction in poultry and swine. Different panels of nature identical compounds, medium chain fatty acids, and plant extracts were employed against strains of Salmonella Typhimurium, Brachyspira hyodysenteriae, and Salmonella Enteritidis, respectively. When bacterial field strains were tested, the comparison between natural compounds and antibiotics was examined, with the aim of evaluating the role of the substances in the antibiotic-resistance context. Results demonstrate that bioactive compounds have positive effects on the host, the pathogen, or both in different experimental conditions. Additionally, when compared to antibiotics, bioactive compounds have proven to be valid alternatives to address the phenomenon of antibiotic resistance.

Food safety and zoonotic enteric pathogens: sources, risk factors and transmission routes of human salmonellosis and campylobacteriosis

Salmonella and Campylobacter are common causes of human gastroenteritis. Their epidemiology is complex and a multi-tiered approach to control is needed, taking into account the different reservoirs, pathways and risk factors. In this thesis, trends in human gastroenteritis and food-borne outbreak notifications in Italy were explored. Moreover, the improved sensitivity of two recently-implemented regional surveillance systems in Lombardy and Piedmont was evidenced, providing a basis for improving notification at the national level. Trends in human Salmonella serovars were explored: serovars Enteritidis and Infantis decreased, Typhimurium remained stable and 4,[5],12:i:-, Derby and Napoli increased, suggesting that sources of infection have changed over time. Attribution analysis identified pigs as the main source of human salmonellosis in Italy, accounting for 43–60% of infections, followed by Gallus gallus (18–34%). Attributions to pigs and Gallus gallus showed increasing and decreasing trends, respectively. Potential bias and sampling issues related to the use of non-local/non-recent multilocus sequence typing (MLST) data in Campylobacter jejuni/coli source attribution using the Asymmetric Island (AI) model were investigated. As MLST data become increasingly dissimilar with increasing geographical/temporal distance, attributions to sources not sampled close to human cases can be underestimated. A combined case-control and source attribution analysis was developed to investigate risk factors for human Campylobacter jejuni/coli infection of chicken, ruminant, environmental, pet and exotic origin in The Netherlands. Most infections (~87%) were attributed to chicken and cattle. Individuals infected from different reservoirs had different associated risk factors: chicken consumption increased the risk for chicken-attributed infections; animal contact, barbecuing, tripe consumption, and never/seldom chicken consumption increased that for ruminant-attributed infections; game consumption and attending swimming pools increased that for environment-attributed infections; and dog ownership increased that for environment- and pet-attributed infections. Person-to-person contacts around holiday periods were risk factors for infections with exotic strains, putatively introduced by returning travellers.

Ocratossina A, metalli, insetticidi e fitoestratti: valutazione degli effetti e degli aspetti residuali nel pollame e in alimenti destinati all'uomo.

La presente tesi di Dottorato di Ricerca descrive le attività di ricerca riguardanti diversi ambiti della sicurezza alimentare, della farmacologia e tossicologia veterinaria. Sono presentati i dati relativi alla ricerca di ocratossina A (OTA) in campioni di bile, rene e fegato di polli, salami artigianali e diversi tipi di formaggio; le concentrazioni di diversi metalli pesanti in uova di galline rurali e 5 tipologie di uova industriali; i dati preliminari ottenuti dalle analisi dei primi campioni di uova e penne provenienti da allevamenti amatoriali di pollame, analizzati per la ricerca del fipronil e metaboliti. Sempre per fipronil, e la molecola foxim, sono presentati dati inerenti a studi di potenziale farmaco-resistenza di popolazioni naturali di Dermanyssus gallinae, e dell’attività insetticida di alcuni fitocomposti a base di tannini. Riguardo all’uso di fitoestratti, sono presentati i dati preliminari ottenuti nei primi mesi di una prova sperimentale in galline di razza Livornese allevata con metodo free-range, finalizzata allo studio della attività anti-infiammatoria e immuno-stimolante, nonché alla valutazione della sicurezza della integrazione nella dieta di Boswellia serrata e Salix alba. Sono stati condotti anche studi traslazionali con altri settori scientifici, relativamente la prevalenza sierologica e microbiologica per Salmonella enterica var. enteritidis e var. thiphymurium in allevamenti ornamentali e da auto-consumo di pollame, e di prevalenza e caratterizzazione di potenziali varianti virali e batteriche (incluse zoonosiche) di alcune malattie infettive del pollame ornamentale. In specie avicole ornamentali è riportato anche uno studio di campo sul trattamento off-label con Fenbendazolo e Flubendazolo in infestazioni massive da Cyathostoma bronchialis in oche esotiche ornamentali. A seguito di una collaborazione con l’Università di Firenze, sono presentati anche i risultati ottenuti da uno studio su modelli murini, dell’azione e funzionamento dei recettori Beta3, espressi nelle neoplasie più frequenti che si verificano nelle donne in gravidanza.