Digestive Enzymes in the Crayfish Cherax albidus: Polymorphism and Partial Characterization

We will deal with the partial characterization and the activity of the following digestive enzymes: amylase, pectinase, alginase, lipase, and protease present in the digestive tract of juvenile freshwater crayfishCherax albidus. Gastric juices, the hepatopancreas, and the intestine were sampled for enzyme analysis. Among carbohydratases, amylase activity was the highest. It was significantly higher in the intestine than in the gastric juice and hepatopancreas. Lipase activity was significantly higher in the hepatopancreas and the intestine compared to the gastric juice. Both alkaline and acid proteases were found. Alkaline proteases were characterized by employing specific protease inhibitors. Both trypsin and chymotrypsin activity was detected. The majority of alkaline protease activity was ascribable to trypsin. Several isoforms of digestive enzymes were identified by gel electrophoresis. This work provides basic information to study the digestive abilities of crayfish

A20 is a negative regulator of BCL10- and CARMA3-mediated activation of NF-κB

The molecular complex containing CARMA proteins, BCL10 and TRAF6 has been identified recently as a key component in the signal transduction pathways that regulate activation of the nuclear factor κB (NF-κB) transcription factor. Here, we report that the inducible protein A20 negatively regulates these signaling cascades by means of its deubiquitylation activity. We show that A20 perturbs assembly of the complex containing CARMA3, BCL10 and IKKγ/NEMO, thereby suppressing activation of NF-κB. Together, our results further define the molecular mechanisms that control activation of NF-κB and reveal a function for A20 in the regulation of CARMA and BCL10 activity in lymphoid and non-lymphoid cells

Development of Biopolymers as Binders for Feed for Farmed Aquatic Organisms

Feed for farmed aquatic organisms should guarantee reasonable degree of stability in aquatic medium, long enough for organisms to consume it. Indeed, there is a general complaint about the loss of nutritional and attractive properties upon exposure of a diet to water. Natural substances able to generate firm pellet feed are therefore highly sought. Such binders should improve the stability of feed, be low cost, and hopefully make and extra source of energy. Biopolymers, have properties that make them suitable for use as aquatic feed binders. Some biopolymers are derived from biomass, biodegradable and renewable. The purpose of this chapter is therefore to review recent reports and progress surrounding the possible employment of sustainable biopolymers as binders for feed for farmed aquatic organisms

Influence of polyphenols from olive mill wastewater on the gastrointestinal tract, alveolar macrophages and blood leukocytes of pigs

In the last years, great importance has been given to the beneficial effects of polyphenols. Among the most relevant health promoting effects, there is the capacity to reduce the amount of free radicals and stimulate the immune response. In this study, polyphenols extracted from olive mill wastewater (OMWW), were fed to adult 'Casertana' pigs during the finishing period. No significant differences in the length of the jejunum-ileum villi and the depth of the colon crypts were detected between control and polyphenols fed pigs. Instead, intra-epithelial and lamina propria leukocytes were more abundant in pigs fed polyphenols (p < .05). Cyclooxygenase-2 immunoreactivity in the gastrointestinal tract, employed as marker of inflammation, was more intense in the control group. Superoxide anion production in primary cell cultures of both blood leukocytes and alveolar macrophages was lower in pigs fed polyphenols (p < .05). Taken together these data indicate that, according to our in vitro studies, OMWW polyphenols seem to be potent antioxidants, while the interpretation of the in vivo experiments is more problematic and further studies are necessary on the interactions between bioactive feed compounds and intestinal status. Such studies can contribute to a better understanding of both positive and negative interactions in vivo and to the identification of new functional feeds.HIGHLIGHTS The effects of polyphenols extracted from olive mill wastewater (OMWW) have been studied in pigs. Gut morphology, inflammation and immune response were investigated. OMWW polyphenols act as potent antioxidants

Breed and Feeding System Impact the Bioactive Anti-Inflammatory Properties of Bovine Milk

In the present study, we aimed at assessing the influence of breed and feeding system on the bovine milk profile of betaines and carnitines and milk capacity in counteracting the inflammatory endothelial cell (EC) damage induced by interleukin (IL)-6. In the first experimental design, two breeds were chosen (Holstein vs. Modicana) to investigate the biomolecule content and antioxidant capacity in milk and dairy products. In the second experimental design, two feeding systems (pasture vs. total mixed ratio) were tested only in Holstein to evaluate the possible effect on the functional profile of milk and dairy products. Finally, the bulk milk from the two experimental designs was used to evaluate the efficacy of preventing IL-6-induced endothelial inflammatory damage. Results showed that Modicana milk and whey had higher biomolecule content and antioxidant activity compared to Holstein milk (p < 0.01). Milk from Holstein fed TMR showed higher concentration of γ-butyrobetaine, δ-valerobetaine (p < 0.01), and l-carnitine (p < 0.05). Similarly, whey from Holstein fed TMR also showed higher content of δ-valerobetaine, glycine betaine, l-carnitine, and acetyl-l-carnitine (p < 0.01) compared to the Holstein fed pasture. Conversely, the antioxidant activity of milk and dairy products was not affected by the feeding system. In ECs, all milk samples reduced the IL-6-induced cytokine release, as well as the accumulation of reactive oxygen species (ROS) and the induction of cell death, with the most robust effect elicited by Modicana milk (p < 0.01). Overall, Modicana milk showed a higher content of biomolecules and antioxidant activity compared to Holstein, suggesting that the breed, more than the feeding system, can positively affect the health-promoting profile of dairy cattle milk

Plasma fatty acid profile in Italian Holstein-Friesian dairy cows supplemented with natural polyphenols from the olive plant Olea Europaea L

This study evaluated the effects of supplementing with natural functional feed on the plasma fatty acid profile of lactating Italian Holstein-Friesian dairy cows. Thirty cows in mid-lactation received the natural olive extract PHENOFEED DRY (500 mg/cow/day) which mainly comprises hydroxytyrosol, tyrosol and verbascoside. The total content of polyphenols and the antioxidant power of standard feed, enriched feed and pure extract was evaluated respectively by Folin-Ciocalteu and DPPH assay, and a characterization in HPLC-UV (High-Perfor mance Liquid Chromatography-Ultraviolet) of bioactive molecules present in the extract PHENOFEED DRY was performed. PHENOFEED DRY was provided for 60 days, and the plasma profile of fatty acids was determined by Gas Chromatography. The administration of enriched feed resulted in an increase in the ratio of Omega-6 to Omega-3 polyunsaturated fatty acids from 3:1 to 4:1 (p<0.001). This was not influenced by the calving order. The addition of polyphenols helped to keep monounsaturated (MUFA) and saturated (SFA) levels constant and results in a significant increase in polyunsaturated (PUFA) fatty acid after 15 days of administration. The Omega 6/Omega-3 ratio was in the optimal range. The findings show that inclusion of natural functional food such as plant polyphenols helps to maintain a healthy blood fatty acid profile in lactating dairy cow

Overlapping distribution of orexin and endocannabinoid receptors and their functional interaction in the brain of adult zebrafish

Hypocretins/Orexins neuropeptides are known to regulate numerous physiological functions, such as energy homeostasis, food intake, sleep/wake cycle, arousal and wakefulness, in vertebrates. Previous studies on mice have revealed an intriguing orexins/endocannabinoids (ECs) signaling interaction at both structural and functional levels, with OX-A behaving as a strong enhancer of 2-arachydonoyl-glycerol (2-AG) biosynthesis. In this study, we describe, for the first time in the brain of zebrafish, the anatomical distribution and co-expression of orexin (OX-2R) and endocannabinoid (CB1R) receptors, suggesting a functional interaction. The immunohistochemical colocalization of these receptors by confocal imaging in the dorsal and ventral telencephalon, suprachiasmatic nucleus (SC), thalamus, hypothalamus, preoptic area (PO) and cerebellum, is reported. Moreover, biochemical quantification of 2-AG levels by LC-MS supports the occurrence of OX-A-induced 2-AG biosynthesis in the zebrafish brain after 3 h of OX-A intraperitoneal (i.p.; 3 pmol/g) or intracerebroventricular (i.c.v.; 0.3 pmol/g) injection. This effect is likely mediated by OX-2R as it is counteracted by i.p./i.c.v administration of OX-2R antagonist (SB334867, 10 pmol/g). This study provides compelling morphological and functional evidence of an OX-2R/CB1R signaling interaction in the brain of adult zebrafish, suggesting the use of this well-established vertebrate animal model for the study of complex and phylogenetically conserved physiological functions

Orexins and receptor OX2R in the gastroenteric apparatus of two teleostean species: Dicentrarchus labrax and Carassius auratus

Orexin A and B peptides and the receptor OX2R were studied in sea bass and goldfish gastroenteric tract by immunoblotting combined with densitometric analysis using NIH Image J software and immunohistochemical techniques. These teleost species present a different gut organization and diverse feeding habits. Immunoblotting experiments showed one band of 16 kDa corresponding to prepro-orexin, and one band of 38 kDa corresponding to the OX2R receptor. Immunohistochemical localization of OXA and OXB was observed in the enteric nervous system throughout the gastroenteric tract of both species. OXA and OXB immunoreactive cells were found in the gastric and intestinal regions of sea bass, and were mainly found in the basal region of folds in intestinal bulb, and in the midgut and hindgut of goldfish. The distribution of OX2R was mainly detected in the mucosa of the gastroenteric tract of sea bass and goldfish. This distribution suggests an endocrine action of OXA and OXB in the gastrointestinal tract as well as involvement in the peripheral control of food intake and digestive processes in both species. This study might also serve to determine the productive factors in breeding and as a baseline for future experimental studies on the regulation of the gastroenteric functions in non-mammalian vertebrates

Chestnut shell tannins: effects on intestinal inflammation and dysbiosis in zebrafish

The aim of the present study was to test the possible ameliorative efficacy of phytochemicals such as tannins on intestinal inflammation and dysbiosis. The effect of a chestnut shell (Castanea sativa) extract (CSE) rich in polyphenols, mainly represented by tannins, on k-carrageenan-induced intestinal inflammation in adult zebrafish (Danio rerio) was tested in a feeding trial. Intestinal inflammation was induced by 0.1% k-carrageenan added to the diet for 10 days. CSE was administered for10 days after k-carrageenan induced inflammation. The intestinal morphology and histopathology, cytokine expression, and microbiota were analyzed. The k-carrageenan treatment led to gut lumen expansion, reduction of intestinal folds, and increase of the goblet cells number, accompanied by the upregulation of pro-inflammatory factors (TNFα, COX2) and alteration in the number and ratio of taxonomic groups of bacteria. CSE counteracted the inflammatory status enhancing the growth of health helpful bacteria (Enterobacteriaceae and Pseudomonas), decreasing the pro-inflammatory factors, and activating the anti-inflammatory cytokine IL-10. In conclusion, CSE acted as a prebiotic on zebrafish gut microbiota, sustaining the use of tannins as food additives to ameliorate the intestinal inflammation. Our results may be relevant for both aquaculture and medical clinic field