Potential of a mixture of eugenol and garlic tincture to improve performance and intestinal health in broilers under necrotic enteritis challenge.

Plant extracts (PE) are gaining increased attention as potential alternatives to in-feed antimicrobials (AM) due to their known antimicrobial activities. This study was conducted to examine the potential of PE, a microencapsulated product composed of eugenol and garlic tincture as an alternative to AM-agent on performance and intestinal health in broilers under necrotic enteritis (NE) challenge. A total of 960 day-old mixed-sex Cobb 500 chicks were randomly distributed to 48-floor pens with 6 treatments replicated 8 times with 20 birds each. The 6 treatments were as follows: UC, unchallenged control; CC, challenged control; PE, challenged group plus PE; AM, challenged group plus AM; FAP, challenged group plus a full dose of AM with PE; HAP, challenged group plus a half dose of AM with PE in starter, grower and finisher phases. Birds in the challenged groups were inoculated with Eimeria spp. on d 9 and Clostridium perfringens on d 14. The body weight gain (BWG), feed intake (FI), feed conversion ratio (FCR), and livability of birds were compromised, and intestinal lesions and mortality were increased (P < 0.05) by NE challenge, illustrating a successful clinical NE challenge. Birds fed AM had higher BWG and FI, and lower FCR, mortality, and intestinal lesions compared to the CC group (P < 0.05). Birds fed PE had improved FCR (P < 0.05) and livability (5.8%) in an overall period compared to the CC group. On d 16, PE supplementation reduced ileal lesion scores in only male birds (P < 0.05). Birds fed PE had decreased Eimeria maxima and Eimeria acervulina oocyst counts in caecal content (P < 0.05). Birds fed PE had decreased Escherichia brunetti and total oocyst counts in caecal content, and E. acervulina oocyst counts in ileal content in only female birds (P < 0.05). On d 35, PE supplementation reduced variation of BW in both male and female birds and increased yellowness (b∗ value, 14.4%) in the thigh. These findings suggest the potential of PE supplementation in diets to improve the performance and intestinal health of birds under clinical NE as indicated by improved FCR, livability, uniformity, reduced ileal lesions, oocyst counts and increased skin yellowness. However, the protective effect of PE may not be apparent in the presence of AM in the feed.

A Microencapsulated Mixture of Eugenol and Garlic Tincture Supplementation Mitigates the Effect of Necrotic Enteritis on Intestinal Integrity and Increases Goblet Cells in Broilers.

This study was conducted to examine the effects of a plant extract mixture, a microencapsulated product composed of eugenol and garlic tincture (PE), on intestinal health in broilers under necrotic enteritis (NE) challenge. A total of 960 d-old mixed-sex Cobb 500 chicks were randomly distributed to 48-floor pens housing 20 birds per pen. Six treatments were applied: UC, unchallenged control; CC, challenged control; PE, challenged group plus PE; AM, challenged group plus antimicrobial (AM); FAP, challenged group plus a full dose of AM with PE; HAP, challenged group plus a half dose of AM with PE in starter, grower and finisher phases. Birds in the challenged groups were inoculated with Eimeria spp. on d 9 and Clostridiumperfringens on d 14. On d 16, the CC group had increased serum fluorescein isothiocyanate dextran (FITC-d), reduced villus surface area, goblet cell number, upregulated CLDN1, JAM2 genes and reduced microbial diversity compared to the UC group (p < 0.05). Birds fed PE had reduced FITC-d, increased goblet cell number and Bifidobacterium compared to the CC group (p < 0.05). Birds fed PE had reduced CLDN5 expression in male birds, and Bacteroides spp. in female birds than CC group (p < 0.05). These findings suggest that PE supplementation mitigates the effect of NE by improving the intestinal health of birds.

Consumption of a Natural High-Intensity Sweetener Enhances Activity and Expression of Rabbit Intestinal Na+/Glucose Cotransporter 1 (SGLT1) and Improves Colibacillosis-Induced Enteric Disorders.

Absorption of glucose, via intestinal Na+/glucose cotransporter 1 (SGLT1), activates salt and water absorption and is an effective route for treating Escherichia coli (E. coli)-induced diarrhea. Activity and expression of SGLT1 is regulated by sensing of sugars and artificial/natural sweeteners by the intestinal sweet receptor T1R2-T1R3 expressed in enteroendocrine cells. Diarrhea, caused by the bacterial pathogen E. coli, is the most common post-weaning clinical feature in rabbits, leading to mortality. We demonstrate here that, in rabbits with experimentally E. coli-induced diarrhea, inclusion of a supplement containing stevia leaf extract (SL) in the feed decreases cumulative morbidity, improving clinical signs of disease (p < 0.01). We show that the rabbit intestine expresses T1R2-T1R3. Furthermore, intake of SL enhances activity and expression of SGLT1 and the intestinal capacity to absorb glucose (1.8-fold increase, p < 0.05). Thus, a natural plant extract sweetener can act as an effective feed additive for lessening the negative impact of enteric diseases in animals.

Accurate quantification of metal-glycinates-sulphate complexes and free metals in feed by capillary electrophoresis inductively coupled plasma mass spectrometry.

Traceability of metal-glycinate-sulphate complexes (Metal-GLY) in feed requires specific analysis to differentiate complexes from inorganic forms. A previously described method focused on the quantification of Metal-GLY at one single concentration but not on the quantification of free metal ion forms. The objective of this work was to extend the method to quantify both Metal-GLY and free metal ion forms of various metals at low inclusion levels. A 50/50 w/w mix of corn flour and soybean meal was used as feed. Copper-glycinate(Cu-GLY), Manganese-glycinate (Mn-GLY) and Zinc-glycinate (Zn-GLY) complexes (provided by Pancosma SA) were used for in-feed inclusions. The feed metal background concentrations and species repartitions were assessed. Cu-GLY was spiked on feed at levels matching 5, 15 and 45 mg/kg, corresponding to metal concentrations of 1.2, 3.6 and 10.8 mg/kg. Mn-GLY and Zn-GLY were spiked at 15, 45 and 100 mg/kg, corresponding to 3.3, 9.9, 22 mg/kg Mn and 3.9, 11.7, 26mg/kg Zn, respectively. The water soluble fraction of un-supplemented feed contained 0.06 mg/kg Cu, 0.05 mg/kg Mn and 0.12 mg/kg Zn, with 69.5% of Cu, 33.2% of Mn and 24.3% of Zn being present under free metal ions but 30.4% of Cu being present under Cu-GLY, 66.82% of Mn and 75.7% of Zn being present under Mn-GLY and Zn-GLY, respectively. The supplemented feeds at the 3 tested doses, from the lowest to the highest inclusion levels, contained in total respectively: 1.1, 3.05 and 9.06 mg/kg Cu; 2.99, 8.9 and 18.2 mg/kg Mn; 3.72, 10.9 and 23.4 mg/kg Zn. The M-GLY species recovered by analysis within the different supplemented feeds ranged from 76.26 to 89.32% for Cu-GLY, form 94.5 to 98.51% for Mn-GLY and from 76.05 to 98.96% for Zn-GLY. These results showed that CE-ICP-MS technique can be used to quantify low doses and to measure metal-species repartition between Metal-GLY and free metal ions, when included in feeds. For the first time, this study highlighted that the raw materials used contain Metal-GLY compounds. This raises the question of the occurrence of these compounds within the different raw materials used in feed production that could dramatically affect the way to supplement minerals in animal feed.

In situ single-crystal to single-crystal (SCSC) transformation of the one-dimensional polymer catena-poly[[diaqua(sulfato)copper(II)]-µ2-glycine] into the two-dimensional polymer poly[µ2-glycine-µ4-sulfato-copper(II)].

The one-dimensional coordination polymer catena-poly[diaqua(sulfato-κO)copper(II)]-µ2-glycine-κ(2)O:O'], [Cu(SO4)(C2H5NO2)(H2O)2]n, (I), was synthesized by slow evaporation under vacuum of a saturated aqueous equimolar mixture of copper(II) sulfate and glycine. On heating the same blue crystal of this complex to 435 K in an oven, its aspect changed to a very pale blue and crystal structure analysis indicated that it had transformed into the two-dimensional coordination polymer poly[(µ2-glycine-κ(2)O:O')(µ4-sulfato-κ(4)O:O':O'':O'')copper(II)], [Cu(SO4)(C2H5NO2)]n, (II). In (I), the Cu(II) cation has a pentacoordinate square-pyramidal coordination environment. It is coordinated by two water molecules and two O atoms of bridging glycine carboxylate groups in the basal plane, and by a sulfate O atom in the apical position. In complex (II), the Cu(II) cation has an octahedral coordination environment. It is coordinated by four sulfate O atoms, one of which bridges two Cu(II) cations, and two O atoms of bridging glycine carboxylate groups. In the crystal structure of (I), the one-dimensional polymers, extending along [001], are linked via N-H···O, O-H···O and bifurcated N-H···O,O hydrogen bonds, forming a three-dimensional framework. In the crystal structure of (II), the two-dimensional networks are linked via bifurcated N-H···O,O hydrogen bonds involving the sulfate O atoms, forming a three-dimensional framework. In the crystal structures of both compounds, there are C-H···O hydrogen bonds present, which reinforce the three-dimensional frameworks.

Determination of Zn-, Cu- and Mn-glycinate complexes in feed samples and in-vitro and in-vivo assays to assess their bioaccessibility in feed samples.

A method was developed for the quantification of Zn-, Cu- and Mn-glycinates in supplemented feed samples. The coupling of capillary electrophoresis (CE) with ICP MS detection after purification of the extract by ultrafiltration was shown to be efficient for the quantitative recovery of glycinates. The method developed was then applied to evaluate the bioaccessibility of glycinates using a sequential enzymolysis approach. The data obtained indicated a strong bioaccessibility of each element (79-94%). A new complex was also found to be formed during the digestion process. Bioavailability was then evaluated by analyzing plasma samples of horses supplemented with glycinates-rich feed. Intact glycinates could not be detected in plasma samples but a Cu-containing molecule was found more abundant after CuGly treatment.

Characterization of metal glycinate complexes by electrospray Q-TOF-MS/MS and their determination by capillary electrophoresis-ICP-MS: application to premix samples.

A method was developed for the determination of metal complexes with glycine (glycinates, [M(Gly)(x)(H(2)O)(y)(SO(4))(z)](n), where M denotes Zn, Cu, Mn and Fe) in premix samples used for the preparation of animal feeds enriched in essential trace elements. The method was based on the extraction of the glycinates with 10 mM ammonium acetate (pH 7.4) followed by their determination using capillary electrophoresis with ICP MS detection. The stability of the glycinates in solution was verified by electrospray TOF-MS. Each supplement was shown to be a mixture of complexes, with polymerization degrees ranging from n = 1 to n = 4 (depending on the metal), that were fully or partially dehydrated. The metal glycine complex moiety was found to be preserved during capillary electrophoresis. The detection limits, calculated as three times the standard deviation of the blank plus the blank, were between 0.05 and 0.2 microg mL(-1) (as the metal), and the calibration curves were linear, allowing the analysis of premix samples. Repeatability for glycinate standards was below 12%, and analytical precision was typically within 15%.

Expression of Na+/glucose co-transporter 1 (SGLT1) in the intestine of piglets weaned to different concentrations of dietary carbohydrate.

Na+/glucose co-transporter 1 (SGLT1) transports dietary sugars from the lumen of the intestine into enterocytes. Regulation of this protein is essential for the provision of glucose to the body and, thus, is important for maintenance of glucose homeostasis. We have assessed expression of SGLT1 at mRNA, protein and functional levels in the intestinal tissue of 28 d old piglets weaned onto isoenergetic diets with differing concentrations of digestible carbohydrate (CHO). We show that expression of SGLT1 remains constant when piglets are fed up to 40 % CHO-containing diets. However, there is a significant increase in SGLT1 expression when the CHO content of the diet is>50 %. Morphometric analyses indicate that the increased expression is not due to a trophic effect. It has been proposed that in rat intestine, in response to a high-CHO diet, GLUT2 (the classical basolateral membrane monosaccharide transporter) is translocated to the luminal membrane of enterocytes to absorb excess dietary glucose. We show, using immunohistochemistry and Western blotting with antibodies raised to amino acids in different epitopes of GLUT2, that under all dietary conditions, low to high CHO, GLUT2 is expressed on the basolateral membrane of pig enterocytes. Furthermore, functional studies indicate that there is no uptake of 2-deoxy-D-glucopyranoside, a specific substrate of Na+-independent glucose transporters into brush-border membrane vesicles isolated from the intestines of piglets either maintained on low- or high-CHO diets. Thus, SGLT1 is the major route for absorption of dietary sugars across the luminal membrane of swine enterocytes.

Expression of Na+/glucose co-transporter 1 (SGLT1) is enhanced by supplementation of the diet of weaning piglets with artificial sweeteners.

In an intensive livestock production, a shorter suckling period allows more piglets to be born. However, this practice leads to a number of disorders including nutrient malabsorption, resulting in diarrhoea, malnutrition and dehydration. A number of strategies have been proposed to overcome weaning problems. Artificial sweeteners, routinely included in piglets' diet, were thought to enhance feed palatability. However, it is shown in rodent models that when included in the diet, they enhance the expression of Na+/glucose co-transporter (SGLT1) and the capacity of the gut to absorb glucose. Here, we show that supplementation of piglets' feed with a combination of artificial sweeteners saccharin and neohesperidin dihydrochalcone enhances the expression of SGLT1 and intestinal glucose transport function. Artificial sweeteners are known to act on the intestinal sweet taste receptor T1R2/T1R3 and its partner G-protein, gustducin, to activate pathways leading to SGLT1 up-regulation. Here, we demonstrate that T1R2, T1R3 and gustducin are expressed together in the enteroendocrine cells of piglet intestine. Furthermore, gut hormones secreted by the endocrine cells in response to dietary carbohydrates, glucagon-like peptides (GLP)-1, GLP-2 and glucose-dependent insulinotrophic peptide (GIP), are co-expressed with type 1 G-protein-coupled receptors (T1R) and gustducin, indicating that L- and K-enteroendocrine cells express these taste elements. In a fewer endocrine cells, T1R are also co-expressed with serotonin. Lactisole, an inhibitor of human T1R3, had no inhibitory effect on sweetener-induced SGLT1 up-regulation in piglet intestine. A better understanding of the mechanism(s) involved in sweetener up-regulation of SGLT1 will allow the identification of nutritional targets with implications for the prevention of weaning-related malabsorption.