Recent advances in monogastric animals, two case studies: transgenic modification of laying hens as animal model and multi-criteria evaluation of Nero d'Aspromonte pig

Trial 1 Protein profiles in transfected hen (Gallus gallus domesticus) oviductal secreting cells Abstract Due to the intensive development of novel biopharming applications, there is a need for simplified in vitro verification models prior to in vivo testing. Laying hen has been already applied as an animal bioreactor to produce the therapeutical enzyme in a rare disease called lysosomal acid lipase deficiency. In this study, we aimed to verify how the proteome of the transfected oviduct epithelial cells would be affected by genetic modification with the human exogene. The study was based on a previously developed method to cultivate chicken oviduct epithelial cells (COEC). The typical characteristics of the COEC epithelial cells were retained across the experiments, including that the adipogenic and osteogenic potentials of differentiating COEC were confirmed with the Oil Red O and von Kossa stain. The mean efficiency of nucleofection ranged from 2.6% to 19.7% depending on the cells’ isolation location in the oviduct (upper, infundibulum site, or magnum). The PCR confirmed the incorporation of human interferon alpha2a (hIFNα2a) exogene into the nucleofected COEC but, the production of hIFNα2a protein did not exceed the detection level in this study. The Ovoalbumin was detected in the non-transfected and transfected COEC, which confirmed the normal secreting functions of the cells subject to modification. Proteomics revealed an increase in cell adhesion molecules and collagen molecules suggesting a trigger of some stimulatory mechanism of nonviral transfection for cell growth. The bioinformatic analyses showed that the negative impact of transfection on cells is limited, and no severe detrimental effect or much disordered biochemical pathway was observed. In conclusion, the observations provide new knowledge about the proteomic profile of the manipulated COEC with regard to the retained normal functionality of the cells, which can be informative for avian biopharma research. Trial 2 Multi-criteria evaluation of Nero d’Aspromonte pig Abstract The aim of the study was to evaluate the effects of rearing system and carcass weight on meat quality of Nero d'Aspromonte pigs. Forty-five pigs reared in plein air-rearing system were fed with concentrate and natural pastures, while 35 pigs reared indoors were fed only concentrate, until reaching two targets slaughter live weights (approximately 140 and 160 kg). At slaughter, carcass weight was recorded (n=14 and n=19 for indoor and plein air, respectively), Longissimus thoracis (LT) muscles were sampled and grouped into four groups: PL (plein air and low carcass weight); PH (plein air and high carcass weight); IL (indoor and low carcass weight); IH (indoor and high carcass weight). The contents of cholesterol and vitamin E in muscle were examined. Antioxidant assays including 2,2’-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 1,1-diphenyl-2-picrylhydrazyl (DPPH) and free thiol were also evaluated among groups. In addition, total lipids content and fatty acid profile were quantitated. The nutritional indices and desaturase activities were also calculated. Neither rearing system nor carcass weight affected vitamin E, cholesterol and intramuscular lipid content. Plein air system decreased the ABTS content in LT, meat from high carcass weight showed higher free thiol content. The plein air system increased total saturated fatty acid (SFA) and decreased total polyunsaturated fatty acid (PUFA) content, resulting in a lower PUFA/SFA ratio and Atherogenic index, but improved the n-6/n-3 ratio. LT muscle from heavier carcasses had lower SFA content and higher total monounsaturated fatty acid content, leading to better n-6/n-3 ratio and Atherogenic and Thrombogenic indices. Moreover, plein air-rearing system increased the activities of Δ5+Δ6 (C20:5n-3/C18:3n-3), Δ5+Δ6 (C22:6n-3/C18:3n-3), and Δ6 (22:6n-3/C20:5n-3). High carcass weight showed higher activity of Δ9-C18 and Δ6 (22:6n-3/C20:5n-3). In conclusion, the plein air-rearing system and high carcass weight could improve the meat quality in Nero d’Aspromonte pigs by enhancing fatty acid composition, nutritional indices and desaturase activities.