Aim: In vitro and in vivo challenge studies were undertaken to develop an in-feed additive of microencapsulated propionic, sorbic acids and pure botanicals to control Campylobacter jejuni in broilers at slaughter age. Methods and Results: Organic acids (OA) and pure botanicals were tested in vitro against Camp. jejuni, whereas in vivo, chickens were fed either a control diet, or increasing doses of the additive for 42 days (experiment 1); in the second experiment, chickens received the additive at 0Á1 or 0Á3% from day 0 to 21 or from day 22 to 42. The additive consistently reduced Camp. jejuni caecal counts at any given dose (exp. 1) or inclusion plan (exp. 2). Moreover, it was able to reduce the number of goblet cells and modify mucin glycoconjugates biosynthesis pattern. Conclusions: We developed an additive that was effective in reducing Camp. jejuni in slaughter-age chickens even at low doses (0Á1%). That efficacy was the result of the synergistic action between OA and botanicals. Significance and Impact of the Study: This study provides a strategy to reduce Camp. jejuni in broilers and, as a consequence, to improve the safety of the food chain. Moreover, data suggest that a treatment limited to the last weeks before slaughter would allow to save on inclusion of the additive throughout the whole production cycle.

Menisci represent fundamental structures for the maintenance of knee homeostasis, playing a key role in knee biomechanics. However, their intrinsic regenerative potential is poor. As a consequence, when a lesion occurs and the meniscus is partially removed by surgery, knee mechanics is subject to dramatic changes. These have been demonstrated to lead often to the development of early osteoarthritis. Therefore, menisci should be repaired whenever possible. In the last decades, tissue engineering approaches have been advocated to improve the reparative processes of joint tissues. In this study, the bonding capacity of an articular chondrocytes-fibrin glue hydrogel was tested as a biologic glue to improve the bonding between two swine meniscal slices in a nude mouse model. The composites were wrapped with acellular fibrin glue and implanted in subcutaneous pouches of nude mice for 4 weeks. Upon retrieval, a firm gross bonding was observed in the experimental samples while none of the control samples, prepared with acellular fibrin glue at the interface, presented any sign of bonding. This was consistent with the histological and scanning electron microscope findings. In particular, a fibrocartilaginous tissue was found at the interface between the meniscal slices, partially penetrating the native meniscus tissue. In order to overcome the lack of regenerative properties of the meniscus, the rationale of using cellular fibrin glue is that fibrin provides immediate stability while carrying cells in the site of lesion. Moreover, fibrin gel is recognized as an optimal scaffold for cell embedding and for promoting fibrocartilaginous differentiation of the cells which synthesize matrix having healing property. These results demonstrated the potential of this model for improving the meniscal bonding. However, further orthotopic studies in a large animal model are needed to evaluate its potential for clinical application.

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 3 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 3 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.

The neuronal and glial changes described here illustrate plasticity of the ENS in response to an altered luminal environment in the gastrointestinal tract.

Research in tissue engineering has been focused on articular cartilage repair for more than a decade. Some pioneristic studies involved the use of hydrogels such as alginate and fibrin glue which still possess valuable potential for cartilage regeneration. One of the main issues in cartilage tissue engineering is represented by the ideal maturation of the construct, before in vivo implantation, in order to optimize matrix quality and integration. The present study was focused on the effect of in vitro culture on a fibrin glue hydrogel embedding swine chondrocytes. We performed an evaluation of the immunohistochemical and biochemical composition and of the biomechanical properties of the construct after 1 and 5 weeks of culture. We noticed that chondrocytes survived in the fibrin glue gel and enhanced their synthetic activity. In fact, DNA content remained stable, while all indices of cartilage matrix production increased (GAGs content, immunohistochemistry for collagen II and safranin-o staining). On the other hand, the biomechanical properties remained steady, indicating a gradual substitution of the hydrogel scaffold by cartilaginous matrix. This demonstrates that an optimal preculture could provide the surgeon with a better engineered cartilage for implantation. However, whether this more mature tissue will result in a more efficient regeneration of the articular surface still has to be evaluated in future investigations.

The aim of this study was to compare low doses of microencapsulated v. pharmacological ZnO in the diet of piglets on growth performance, ileum health status and architecture. One hundred and forty-four piglets weaned at 28 days and divided in 36 pens (two males and two females per pen), received a basal diet (control, Zn at 50 mg/kg) or the basal diet with ZnO at 3000 mg/kg (pZnO), or with lipid microencapsulated ZnO at 150 or 400 mg/kg (mZnO-300 and mZnO-800, respectively). After 14 and 42 days, three pigs per sex per treatment were euthanized to collect the ileum mucosa for immunohistochemistry, histomorphology, inflammatory cytokines and tight junction components gene expression. Data were analyzed with one-way ANOVA. At 0 to 14 days, the pZnO and mZnO-800 groups had greater average daily gain compared with control ( P < 0.05). Gain to feed ratio (G : F) in the same time interval was higher in pZnO group compared with control thus resulting in higher BW ( P < 0.05). At day 14, ileum villi height in mZnO-800 pigs was 343 µm v. 309 and 317 µm in control and pZnO, respectively ( P < 0.01) and villi : crypts ratio (V : C), as well as cells positive to proliferating cell nuclear antigen (PCNA), were greater in all treated groups compared with control ( P < 0.01). In mZnO-800 group, interferon-γ mRNA was the lowest ( P = 0.02), and both pharmacological ZnO and mZnO reduced tumor necrosis factor-α protein level ( P < 0.0001). Compared with pZnO group, mZnO-800 increased occludin and zonula occludens-1 protein level (1.6-fold and 1.3-fold, respectively; P < 0.001). At day 42, both groups receiving microencapsulated ZnO had 1.7 kg greater BW than control and did not differ from pZnO group ( P = 0.01); ileum villi height and V : C ratio were the greatest for pZnO compared with the other groups, whereas PCNA-positive cells were the most numerous in mZnO-800 group ( P < 0.001).In conclusion, pigs receiving low doses of microencapsulated ZnO had G : F comparable with those receiving pharmacological level of ZnO in the overall post-weaning phase. Moreover, in the first 2 weeks post-weaning, microencapsulated ZnO effect on inflammatory status and ileum structure and integrity was comparable with pharmacological ZnO.

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.

The present study evaluated the effects of a novel plant extract (PE) product (Grazix TM ) on the performance and gut health of weaned piglets challenged with Escherichia coli. The PE was a standardised mixture of green tea leaves (Camellia sinensis) and pomegranate fruit (Punica granatum) obtained by using the LiveXtract TM process. A total of 144 piglets were weaned at 24 days and allocated to 8 for a 35-day experiment with a 2 × 2 × 2 factorial design comparing different treatments (water without product (CT) or 8 μl/kg per day PE in drinking water (PE)), feeding regimens (ad libitum (AD) or restricted (RE)) and oral E. coli challenges on day 9 (sham (− ) or infected ( + )). There were six pens per group with three piglets per pen. On day 35, 24 of the RE feeding piglets were slaughtered. It was found that PE supplementation increased the average daily gain (ADG) from day 28 to day 35 ( P = 0.03) and increased the gain to feed ratio (G : F) from day 7 to day 14 ( P = 0.02). RE feeding led to lower feed intake in piglets during the 1 st week ( P < 0.01), 2 nd week ( P = 0.06), 3 rd week ( P = 0.05), and throughout the course of the overall study period ( P = 0.05). E. coli challenge decreased the ADG and G : F ratio from day 7 to day 14 ( P = 0.08 and < 0.01, respectively) and increased the faecal score (higher values indicate more severe diarrhoea) on days 14, 21, 28 and 35 ( P < 0.01). PE supplementation decreased the faecal score in the challenged piglets during the 1 st week post-challenge ( P < 0.01). E. coli challenge increased the faecal E. coli level on day 14 ( P = 0.03) and increased the Enterobacteriaceae level on day 35 ( P < 0.01). Reduced faecal E. coli was observed on days 14 and 35 ( P = 0.05 and 0.02, respectively), and reduced Enterobacteriaceae ( P < 0.01) was found on day 35 in the PE animals. RE feeding increased the faecal Lactobacillus, Enterobacteriaceae and E. coli levels on day 35 ( P = 0.02, < 0.01 and < 0.01, respectively). These results suggest that PE supplementation may improve the gut health status of post-weaning piglets and counteract some of the negative effects that occur when piglets are challenged with E. coli.Keywords: Escherichia coli, gut health, performance, piglet, plant extract ImplicationsThis study investigated the effects of a novel plant extract product that was added to the drinking water for weaned piglets. Different feeding regimens were tested, and the effects of the extract on growth performance, gut health and protection against Escherichia coli challenge were observed. The results showed that the plant extract may represent a useful additive for improving gut health and microbial ecology and reducing the severity of an E. coli challenge. The results may have a significant impact on nutritional management in conventional farms. Use of the plant extract could enable piglets to better withstand the infections that are often associated with weaning.