C. Domeneghini

Localization of IGF-I, IGF-I receptor, and IGFBP-2 in developing Umbrina cirrosa (Pisces: Osteichthyes).

In this study, the distribution of IGF-I, IGF type I receptor (IGF-IR), and IGF-binding protein 2 (IGFBP-2) was investigated during larval and post-larval developmental stages of the shi drum (Umbrina cirrosa) by immunohistochemistry using antisera raised against Sparus aurata IGF-I and IGF-IR, and against mouse IGFBP-2. Immunoreactivity of the mitogenic marker PCNA (proliferating cell nuclear antigen) was used for assessment of cellular proliferation. Distribution of IGF-I mRNA was studied by in situ hybridization. IGF-I immunoreactivity was detected in liver and developing intestine already in 1-5 day post-hatching larvae. From day 11, immunostaining in the intestine was evident in the enterocytes of the anterior intestine and in the apical zone of the epithelium of developing posterior intestine. Positive reaction with IGF-I antibody was also detected in chondrocytes, in the epithelium of the skin, gills and in the central nervous system (CNS), and lateral muscle. At hatching IGF-IR immunoreactivity was already detectable in developing CNS, notochord, and skin. From day 6 immunostaining was evident in the olfactory epithelium, in eyes and from day 11 in the developing olfactory bulbs and CNS. Positive reaction with IGF-IR antibody was also detected in chondrocytes, in the epithelium of the skin, gills, heart, and in the lateral muscle. Immunoreactive IGFBP-2, as detected by anti-mouse IGFBP-2 antiserum, exhibited generally a similar distribution pattern to that of IGF-I and IGF-IR. In situ hybridization, which has been performed by using riboprobes from S. aurata cDNA, revealed IGF-I mRNA in skeletal musculature, liver, and CNS. These data strongly suggest a role for the IGF system during development and growth of U. cirrosa.

Influence of dietary conjugated linoleic acids and vitamin E on meat quality, and adipose tissue in rabbits

Eighty New Zealand White rabbits, age 55 days, half male and half female, were randomly assigned to one of four diets in a 2×2×2 factorial arrangement. The diets contained 0% or 0.5% CLA, and 60 or 240mg/kg α-Tocopheryl acetate. Forty-eight rabbits were slaughtered at age 92 days. Growth performances and carcass weight were higher (P<0.05) in 240mg/kg of α-tocopheryl acetate-fed rabbits. Fat and CLA isomers content of Longissimus Lumborum (LL) muscle was higher (P<0.05) in CLA-fed rabbits than control. Fatty acid composition of LL muscle was modified (P<0.05) and oxidative stability was increased (P<0.001) by both dietary treatments. CLA increased (P<0.05) triglyceride, total cholesterol and glucose levels in plasma. Adipocytes in interscapular and perirenal fat in the 240mg/kg α-tocopheryl acetate and 0.5% CLA groups were larger in size but lesser in number than in 60mg/kg α-tocopheryl acetate and no CLA rabbit (P<0.01).

The chemical code of porcine enteric neurons and the number of enteric glial cells are altered by dietary probiotics

Background  The enteric nervous system (ENS) contains chemically coded populations of neurons that serve specific functions for the control of the gastrointestinal tract. The ability of neurons to modify their chemical code in response to luminal changes has recently been discovered. It is possible that enteric neuronal plasticity may sustain the adaptability of the gut to changes in intestinal activity or injury, and that gut neurons may respond to an altered intestinal environment by changing their neuropeptide expression.

Can nutraceuticals affect the structure of intestinal mucosa? Qualitative and quantitative microanatomy in L-glutamine diet-supplemented weaning piglets

Weaning piglets were fed an L -glutamine-supplemented diet with the aim of monitoring the effects on gut mucosal turnover and barrier function, to elucidate the potential preventive or therapeutic roles of glutamine as a nutraceutical or ‘functional food’. Sixteen female weaning piglets were divided into two groups, which were fed a control diet (Ctr group: n = 8) or a Ctr + 0.5% L -glutamine diet (G group: n = 8) for 28 days. In the ileum of group G piglets the villus height (V) and crypt depth (C) were increased, and the V:C ratio was decreased (p < 0.01). The PCNA and TUNEL immunoreactivities were also tested. The number of mitotic mucosal cells (M) was increased, and that of mucosal cells with apoptotic nuclei (A) decreased in such a way that the A:M index diminished (p < 0.01). The A:M index also decreased at the level of some components of the gut-associated lymphatic tissue (GALT), thus indicating a positive effect on the gut barrier function. This trial showed that L -glutamine supplementation influenced some morphofunctional characteristics of piglet ileal mucosa. These data corroborate the nutraceutical role of glutamine as a trophic agent for mucosal repair, improvement of barrier function and gut adaptation in the swine per se and as an animal model.

Role of autologous rabbit adipose‐derived stem cells in the early phases of the repairing process of critical bone defects

Adipose‐derived stem cells (ASCs) may represent a novel and efficient tool to promote bone regeneration. In this study, rabbit ASCs were expanded in culture and used for the regeneration of full‐thickness bone defects in the proximal epiphysis of tibia of 12 New Zealand rabbits. Defects were implanted with graft material as follows: untreated (control), empty hydroxyapatite (HA) disk, ASCs alone, and HA disk seeded with ASCs. Each isolated ASCs population was tested in vitro: they all showed a high proliferation rate, a marked clonogenic ability, and osteogenic differentiation potential. Eight weeks after implantation, macroscopic analyses of all the samples showed satisfactory filling of the lesions without any significant differences in term of stiffness between groups treated with or without cells (p > 0.05). In both the scaffold‐treated groups, a good osteointegration was radiographically observed. Even if HA was not completely reabsorbed, ASCs‐loaded HA displayed a higher scaffold resorption than the unloaded ones. Histological analyses showed that the osteogenic abilities of the scaffold‐treated defects was greater than those of scaffold‐free samples, and in particular new formed bone was more mature and more similar to native bone in presence of ASCs. These results demonstrated that autologous ASCs–HA constructs is a potential treatment for the regeneration of bone defects.

Effects of pig genotype (Iberian v. Landrace × Large White) on nutrient digestibility, relative organ weight and small intestine structure at two stages of growth.

Although the effects of pig genotype on total-tract apparent digestibility (TTAD) have been widely reported in the literature, there is controversial information on the digestive capacity of indigenous breeds compared with lean-type pigs. The strategy of this study was to test the effects of pig genotype and crude protein (CP) supply on performance, digestive utilization of nutrients, relative organ weight and morphometric analysis of the small intestine. Thirty-eight Iberian (IB) and Landrace × Large White (LD) pigs were used. Three pigs per genotype were slaughtered at approximately 15 kg BW. The remaining pigs were fed one of two diets differing in CP content (13% or 17% as fed) using a pair-fed procedure. Feeding level was restricted at 0.8 × ad libitum of IB pigs. Nutrient digestibility and nitrogen (N) balance trials were performed at 30 and 80 kg BW. Four pigs per dietary treatment and genotype were slaughtered at approximately 50 and 115 kg BW. The gastrointestinal tract and the rest of the visceral organs were weighed and samples of the small intestine were taken to carry out histological and histometrical studies. Daily gain and gain-to-feed ratio were higher in LD than in IB pigs during the fattening and growing-fattening periods (P < 0.01). N TTAD was significantly higher for LD pigs at 30 kg BW (P < 0.05), whereas at 80 kg BW we observed greater values for digestibility of organic matter and energy in IB pigs (averaging 1.5%, P < 0.01). Both N retention (NR) and efficiency of NR were increased in LD pigs at 30 and 80 kg BW (30% as mean value). The proportional weight of the small intestine was greater in LD than in IB pigs at 50 and 115 kg BW. Histometry showed that IB presented a lower muscle layer thickness than LD pigs in ileum, irrespective of the BW (P < 0.05). In contrast, LD pigs showed approximately 10% higher ileal villi length and villi-to-crypt ratio than IB pigs at 115 kg BW. CP supply affected to a larger extent the small intestinal micro-anatomical structure of LD pigs at 50 kg BW. In conclusion, our results suggests that although the higher growth rate, NR and efficiency of NR observed in LD pigs might be associated with presumably more efficient structural aspects of the small intestine, the main differences between the two genotypes should be attributed to a larger extent to protein and energy utilization in tissues with consequences for the overall efficiency of energy use.

A tissue engineered osteochondral plug : an in vitro morphological evaluation

Articular cartilage lesions have a poor intrinsic healing potential. The repair tissue is often fibrous, having insufficient biomechanical properties, which could frequently lead to the development of early osteoarthritis. In the last decade, tissue engineering approaches addressed this topic in order to restore joint function with a differentiated and functional tissue. Many biomaterials and techniques have been proposed and some of them applied in clinical practice, even though several concerns have been raised on the quality of the engineered tissue and on its integration in the host joint. In this study, we focused on engineering in vitro a biphasic composite made of cellular fibrin glue and a calcium–phosphate scaffold. Biphasic composites are the latest products of tissue engineering applied to articular cartilage and they seem to allow a more efficient integration of the engineered tissue with the host. However, a firm in vitro bonding between the two components of the composite is a necessary condition to validate this model. Our study demonstrated a gross and microscopic integration of the two components and a cartilage-like quality of the newly formed matrix. Moreover, we noticed an improvement of this integration and GAGs production during the in vitro culture.

Ontogeny of the digestive tract in sharpsnout sea bream Diplodus puntazzo (Cetti, 1777).

The ontogeny of the digestive tract was studied histologically and histochemically in sharpsnout sea bream Diplodus puntazzo from hatching (0 DAH, Days After Hatching) until day 57 (57 DAH). At hatching, the digestive tract appeared as a histologically undifferentiated straight tube lying dorsally to the yolk sac. When the mouth opened at 3 DAH, the digestive tract was differentiated into buccopharynx, oesophagus, incipient stomach and intestine. The pancreas, liver and gall bladder were also differentiated at this stage and both the bile and pancreatic duct had opened into the anterior intestine. Active feeding began in 50% of larvae at 4 DAH, although permanence of yolk reserves until 7 DAH suggests a period of both endogenous and exogenous feeding. Nutrient absorption was first visible from 5 DAH, as colourless supra- and infranuclear vacuoles in the anterior intestinal mucosa, suggesting a lipid content, as well as supranuclear, eosinophilic vacuoles, containing protein, in the posterior intestinal mucosa. Early caecal development could be detected from 10 DAH, whereas gastric glands appeared at 30 DAH, indicating the transition from larval to juvenile stage and the acquisition of an adult mode of digestion. Goblet cells appeared in the digestive tract of sharpsnout sea bream larvae shortly after first feeding. The mucus content of goblet cells varied with the digestive region and, in the buccal cavity and oesophagus, also with the developmental phase. This study provides knowledge for better husbandry practices in the aquaculture industry, as well as for the implementation of future nutritional studies.

Diatom extraction with HCl from animal tissues: A technical note

Identification of diatoms from human or animal tissue may be affected by factors concerning techniques of extraction and analysis which cause fragmentation or loss of diatoms. In this brief technical note a novel efficient method of extraction of diatoms from animal tissues is presented. The authors have tested an H(2)O(2) plus HCl and HCl method to detect diatoms in several tissues from pig immersed in water rich in diatoms; they have also compared this method to other methods of extraction including enzymatic ones. The experiment showed that all siliceous frustules of fresh water diatoms are resistant to the treatment proposed and are still recognizable after the digestion. Among the different tests, treatment by 20% HCl (T1) proved to be more simple and time-saving, and seems to be a valuable diagnostic tool for routine forensic procedures in the extraction and detection of diatoms. The positive results obtained in all cases should lead to further studies in order to evaluate the reliability of the proposed methods also from a quantitative point of view.

Effects of a blend of essential oils and an enzyme combination on nutrient digestibility, ileum histology and expression of inflammatory mediators in weaned piglets

The aim of this study was to test the hypothesis of an improved gut environment of post-weaning piglets when administered a blend of essential oils (EO; thymol and cinnamaldehyde) and an enzyme combination (xylanase and β-glucanase (XB)) either alone or in combination. To assess the effect of dietary treatments, faecal nutrient digestibility and microbial counts, as well as ileum histology and gene expression of inflammatory mediators were evaluated. One hundred and ninety-two weaned piglets were allocated into four experimental treatments, and fed the basal diet (CTRL) either without or with EO, XB or their combination (EO+XB) for a 42-day period. The experiment concerning digestibility was designed with two periods (period I: days 15 to 21; period II: days 29 to 35) and the faeces were collected on days 20, 21, 34 and 35. On day 42, six piglets from each treatment were slaughtered. It was found that EO, XB and EO+XB supplementation did not affect (P>0.05) the growth performance of the piglets from days 0 to 42. Moreover, no dietary effect on faecal score was observed. Faecal digestibility of dry matter, organic matter, ash, dietary fibre, lipid, CP and NDF were increased from period I to period II (P<0.01 to P=0.06), while no effect (P>0.05) of EO, XB or their combination on the faecal digestibility was observed at both periods. Compared with the CTRL diet, dietary XB reduced the faecal Lactobacillus and Escherichia coli counts but increased the Lactobacillus to Coliforms ratio on day 42 (P=0.02, 0.03 and 0.03, respectively), and all the additives supplementations decreased the counts of faecal Coliforms on day 42 (P<0.01). XB supplementation increased the villus to crypt ratio (P=0.04) and reduced the mucosal macrophages number (P<0.01) in the ileum compared with the CTRL group, and dietary EO or EO+XB decreased the number of lymphatic follicles (P=0.01 and P<0.01, respectively) and mucosal macrophages (P=0.02 and P<0.01, respectively). In addition, the interleukin (IL)-1α was downregulated in piglets treated with EO+XB compared with the EO group (P=0.02). In conclusion, the administration of EO, XB or their combination was effective in improving ileum histology, and EO+XB supplementation might benefit the modulation of the expression of ileum inflammatory cytokines in piglets.