Indagine sulla presenza di Helicobacter pullorum in allevamenti avicoli italiani

From September 2005 to December 2006, in order to define the prevalence of Helicobacter pullorum in broiler chickens, laying hens and turkey, a total of 365 caecum contents of animals reared in 76 different farms were collected at the slaughterhouse. A caecum content of a ostrich was also sampled. In addition, with the aim of investigating the occurrence of H. pullorum in humans, 151 faeces were collected at the Sant’Orsola-Malpighi University Hospital of Bologna from patients suffering of gastroenteritis. A modified Steele–McDermott membrane filter method was used. Gram-negative curved rod bacteria were preliminary identified as H. pullorum by a PCR assay based on 16S rRNA, then subjected to a RFLP-PCR assay to distinguish between H. pullorum and H. canadensis. One isolate from each farm was randomly selected for phenotypic characterization by biochemical methods and 1D SDSPAGE analysis of whole cell proteins profiles. Minimum Inhibitory Concentration (MIC) for seven different antibiotics were also determined by agar dilution method. Moreover, to examine the intraspecific genomic variability, two strains isolated from 17 different farms were submitted to genotyping by Pulse-Field Gel Electrophoresis (PFGE). In order to assess the molecular basis of fluorquinolone resistance in H. pullorum, gyrA of H. pullorum CIP 104787T was sequenced and nucleotide sequences of the Quinolone Resistance Determining Region (QRDR) of a total of 18 poultry isolates, with different MIC values for ciprofloxacin and nalidixic acid, were compared. According to the PCR and PCR-RFLP results, 306 out of 366 animals examined were positive for H. pullorum (83,6%) and 96,1% of farms resulted infected. All positive samples showed a high number of colonies (>50) phenotipically consistent with H. pullorum on the first isolation media, which suggests that this microrganism, when present, colonizes the poultry caecum at an elevate load. No human sample resulted positive for H. pullorum. The 1D SDS-PAGE whole protein profile analysis showed high similarity among the 74 isolates tested and with the type strain H. pullorum CIP 104787T. Regarding the MIC values, a monomodal distribution was found for ampicillin, chloramphenicol, gentamicin and nalidixic acid, whereas a bimodal trend was noticed for erythromycin, ciprofloxacin and tetracycline (indicating an acquired resistance for these antibiotics). Applying the breakpoints indicated by the CSLI, we may assume that all the H. pullorum tested are sensitive only to gentamicin. The intraspecific genomic variability observed in this study confirm that this species don’t have a clonal population structure, as motioned by other autors. The 2490 bp gyrA gene of H. pullorum CIP104787T with an Open Reading Frame (ORF) encoding a polypeptide of 829 amino acids was for the first time sequenced and characterized. All ciprofloxacin resistant poultry isolates showed ACA®ATA (Thr®Ile) substitution at codon 84 of gyrA corresponding to codons of gyrA 86, 87 and 83 of the Campylobacter jejuni, H. pylori and Escherichia coli, respectively. This substitution was functionally confirmed to be associated with the ciprofloxacin resistant phenotype of poultry isolates. This is the first report of isolation of H. pullorum in turkey and in ostrich, indicating that poultry species are the reservoir of this potential zoonotic microorganisms. In order to understand the potential role as food-borne human pathogen of H. pullorum, further studies must be carried on.

Effect of rearing techniques and feed composition on productive traits, bird welfare and quality of poultry products

The PhD project was focused on the study of the poultry welfare conditions and improvements. The project work was divided into 3 main research activities. A) Field evaluation of chicken meat rearing conditions kept in intensive farms. Considering the lack of published reports concerning the overall Italian rearing conditions of broiler chickens, a survey was carried out to assess the welfare conditions of broiler reared in the most important poultry companies in Italy to verify if they are in accordance with the advices given in the European proposal COM (2005) 221 final. Chicken farm conditions, carcass lesions and meat quality were investigated. 1. The densities currently used in Italy are in accordance with the European proposal COM 221 final (2005) which suggests to keep broilers at a density lower than 30-32 kg live weight/m2 and to not exceed 38-40 kg live weight/m2. 2. The mortality rates in summer and winter agree with the mortality score calculated following the formula reported in the EU Proposal COM 221 final (2005). 3. The incidence of damaged carcasses was very low and did not seem related to the stocking density. 4. The FPD scores were generally above the maximum limit advised by the EU proposal COM 221 final (2005), although the stocking densities were lower than 30-32 kg live weight per m2. 5. It can be stated that the control of the environmental conditions, particularly litter quality, appears a key issue to control the onset of foot dermatitis. B) Manipulation of several farm parameters, such litter material and depth, stocking density and light regimen to improve the chicken welfare conditions, in winter season. 1. Even though 2 different stocking densities were established in this study, the performances achieved from the chickens were almost identical among groups. 2. The FCR was significantly better in Standard conditions contrarily to birds reared in Welfare conditions with lower stocking density, more litter material and with a light program of 16 hours light and 8 hours dark. 3. In our trial, in Standard groups we observed a higher content of moisture, nitrogen and ammonia released from the litter. Therefore it can be assumed that the environmental characteristics have been positively changed by the improvements of the rearing conditions adopted for Welfare groups. 4. In Welfare groups the exhausted litters of the pens were dryer and broilers showed a lower occurrence of FPD. 5. The prevalence of hock burn lesions, like FPD, is high with poor litter quality conditions. 6. The combined effect of a lower stocking density, a greater amount of litter material and a photoperiod similar to the natural one, have positively influenced the chickens welfare status, as a matter of fact the occurrence of FPD in Welfare groups was the lowest keeping the score under the European threshold of the proposal COM 221 final(2005). C) The purpose of the third research was to study the effect of high or low stocking density of broiler chickens, different types of litter and the adoption of short or long lighting regimen on broiler welfare through the evaluation of their productivity and incidence of foot pad dermatitis during the hot season. 1. The feed efficiency was better for the Low Density than for High Density broilers. 2. The appearance of FPD was not influenced by stocking density. 3. The foot examination revealed that the lesions occurred more in birds maintained on chopped wheat straw than on wood shaving. 4. In conclusion, the adoptions of a short light regimen similar to that occurring in nature during summer reduces the feed intake without modify the growth rate thus improving the feed efficiency. Foot pad lesion were not affected neither by stocking densities nor by light regimens whereas wood shavings exerted a favourable effect in preserving foot pad in good condition. D) A study was carried out to investigate more widely the possible role of 25-hydroxycholecalciferol supplemented in the diet of a laying hen commercial strain (Lohmann brown) in comparison of diets supplemented with D3 or with D3 + 25- hydroxycholecalciferol. Egg traits during a productive cycle as well as the bone characteristics of the layers have been as well evaluated to determine if there the vitamin D3 may enhance the welfare status of the birds. 1. The weight of the egg and of its components is often greater in hens fed a diet enriched with 25-hydroxycholecalciferol. 2. Since eggs of treated groups are heavier and a larger amount of shell is needed, a direct effect on shell strength is observed. 3. At 30 and at 50 wk of age hens fed 25 hydroxycholecalciferol exhibited greater values of bone breaking force. 4. Radiographic density values obtained in the trial are always higher in hens fed with 25-hydroxycholecalciferol of both treatments: supplemented for the whole laying cycle (25D3) or from 40 weeks of age onward (D3+25D3).

Farming and processing stategies for improving poultry meat quality

The purpose of the PhD research was the identification of new strategies of farming and processing, with the aim to improve the nutritional and technological characteristics of poultry meat. Part of the PhD research was focused on evaluation of alternative farming systems, with the aim to increase animal welfare and to improve the meat quality and sensorial characteristics in broiler chickens. It was also assessed the use of innovative ingredients for marination of poultry meat (sodium bicarbonate and natural antioxidants) The research was developed by studying the following aspects: - Meat quality characteristics, oxidative stability and sensorial traits of chicken meat obtained from two different farming systems: free range vs conventional; - Meat quality traits of frozen chicken breast pre-salted using increasing concentrations of sodium chloride; - Use of sodium bicarbonate in comparison with sodium trypolyphosphate for marination of broiler breast meat and phase; - Marination with thyme and orange essential oils mixture to improve chicken meat quality traits, susceptibility to lipid oxidation and sensory traits. The following meat quality traits analyseswere performed: Colour, pH, water holding capacity by conventional (gravimetric methods, pressure application, centrifugation and cooking) and innovative methods (low-field NMR and DSC analysis) ability to absorb marinade soloutions, texture (shear force using different probes and texture profile analysis), proximate analysis (moisture, proteins, lipids, ash content, collagen, fatty acid), susceptibility to lipid oxidation (determinations of reactive substances with thiobarbituric acid and peroxide value), sensorial analysis (triangle test and consumer test).

Ricerca di contaminanti in tessuti animali e in alimenti destinati all'uomo.

La contaminazione chimica rappresenta uno dei rischi principali per la sicurezza alimentare e può arrecare anche gravi danni alla salute umana. Rientrano in questa tesi di dottorato tre famiglie di contaminanti: Micotossine, Metalli e Insetticidi. La ricerca di aflatossina B1 è stata effettuata su 90 confezioni di farina, sia biologici sia convenzionali. La presenza della micotossina è stata rilevata solo nelle farine di mais. Solo un campione di produzione convenzionale ha superato il limite di 2 ppb definito per legge. Il dato di maggior rilievo è stato che il quantitativo di 5 grammi di campionamento si è dimostrato non rappresentativo sul totale della confezione commerciale di farina. Più attendibile si è invece dimostrato un campionamento di 20 grammi. L’aflatossina M1 è stata ricercata in 58 campioni di latte di cui 35 sono risultati positivi. Tuttavia, i livelli riscontrati erano costantemente inferiori al limite previsto per legge. Sono stati sottoposti a estrazione e purificazione, e analizzati con metodica HPLC-FL per la ricerca di Ocratossina A, 114 campioni di bile, 35 campioni di plasma, 40 campioni di rene prelevati da polli in Giordania. Le analisi hanno fornito risultati costantemente negativi. Sono stati analizzati 72 campioni (30 di muscolo, 29 di fegato e 13 di rene) prelevati da 30 bovini nel macello di Irbid (Giordania), di età compresa tra 8 e 30 mesi e provenienti da allevamenti diversi, per la ricerca di 13 elementi essenziali e non essenziali. In questo studio nessun campione supera i livelli massimi stabiliti dalla normativa europea per quanto riguarda gli elementi considerati. Infine, sono stati analizzati 37 campioni di latte ovino e 31 campioni di latte bovino, prelevati in Giordania in diversi allevamenti, per la ricerca di 4 neonicotinoidi (imidacloprid, acetamiprid, thiamethoxam e thiacloprid). I campioni, analizzati con sistema HPLC/MS/MS, sono risultati costantemente negativi ai quattro neonicotinoidi ricercati.

Farming and processing factors for improving quality properties of poultry and rabbit meat

Recently, global meat market is facing several dramatic changes due to shifting in diet and life style, consumer demands, and economical considerations. Firstly, there was a tremendous increase in the poultry meat demand. Furthermore, current forecast and projection studies pointed out that the expansion of the poultry market will continue in future. In response to this demand, there was a great success to increase growth rate of meat-type chickens in the last few decades in order to optimize the production of poultry meat. Accordingly, the increase of growth rate induced the appearance of several muscle abnormalities such as pale-soft-exudative (PSE) syndrome and deep-pectoral-myopathy (DPM) and more recently white striping and wooden breast. Currently, there is growing interest in meat industry to understand how much the magnitude of the effect of these abnormalities on different quality traits for raw and processed meat. Therefore, the major part of the research activities during the PhD project was dedicated to evaluate the different implications of recent muscle abnormalities such as white striping and wooden breast on meat quality traits and their incidence under commercial conditions. Generally, our results showed that the incidence of these muscle abnormalities was very high under commercial conditions and had great adverse impact on meat quality traits. Secondly, there is growing market share of convenient, healthy, and functional processed meat products. Accordingly, the remaining part of research activities of the PhD project was dedicated to evaluate the possibility to formulate processed meat products with higher perceived healthy profile such as phosphate free-marinated chicken meat and low sodium-marinated rabbit meat products. Overall all findings showed that sodium bicarbonate can be considered as promising component to replace phosphates in meat products, while potassium chloride under certain conditions was successfully used to produce low marinated rabbit meat products.