3 results
Intestinal permeability induced by lipopolysaccharide and measured by lactulose, rhamnose and mannitol sugars in chickens
- S. Gilani, G. S. Howarth, S. M. Kitessa, C. D. Tran, R. E. A. Forder, R. J. Hughes
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Increased intestinal permeability (IP) can lead to compromised health. Limited in vivo IP research has been conducted in chickens. The objectives of the current study were to develop a model of increased IP utilizing lipopolysaccharide (LPS Escherichia coli O55:B5) and to evaluate IP changes using the lactulose, mannitol and rhamnose (LMR) sugar permeability test. In addition, fluorescein isothiocyanate dextran (FITC-d), d-lactate, zonula occludens (ZO-1) and diamine oxidase (DAO) permeability tests were employed. Male Ross chickens were reared until day 14 on the floor in an animal care facility and then transferred to individual cages in three separate experiments. In each of experiments 1 and 2, 36 chicks were randomly allocated to receive either saline (control) or LPS (n=18/group). Lactulose, mannitol and rhamnose sugar concentration in blood was measured at 0, 30, 60, 90, 120 and 180 min in experiment 1, at 60, 90 and 120 min in experiment 2 and at 90 min in experiment 3 (n=16/group). Lipopolysaccharide was injected intraperitoneally at doses of 0.5, 1 and 1 mg/kg BW in experiments 1, 2 and 3, respectively, on days 16, 18 and 20, whereas control received sterile saline. On day 21, only birds in experiments 1 and 2 were fasted for 19.5 h. Chicks were orally gavaged with the LMR sugars (0.25 gL, 0.05 gM, 0.05 gR/bird) followed by blood collection (from the brachial vein) as per time point for each experiment. Only in experiment 3, were birds given an additional oral gavage of FITC-d (2.2 mg/ml per bird) 60 min after the first gavage. Plasma d-lactate, ZO-1 and DAO concentrations were also determined by ELISA in experiment 3 (n=10). Administration of LPS did not affect IP as measured by the LMR sugar test compared with control. This was also confirmed by FITC-d and DAO levels in experiment 3 (P>0.05). The plasma levels of d-lactate were decreased (P<0.05). Plasma levels of ZO-1 were increased in the third experiment only and did not change in the first two experiments. Lipopolysaccharide at doses of 0.5 and 1 mg/kg did not increase IP in this model system. In conclusion, the LMR sugar can be detected in blood 90 min after the oral gavage. Further studies are needed for the applicability of LMR sugars tests.
Effects of dose and route of administration of genistein on isoflavone concentrations in post-weaned and gestating sows
- C. Farmer, P. Robertson, G. S. Gilani
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Phytoestrogens could be a useful tool in swine husbandry practices because of their structural and functional similarities to estradiol. The goal of this study was to compare various routes and doses of administration of the phytoestrogen genistein in sows of two different physiological statuses. Circulating concentrations of isoflavones, estradiol and IGF-I were determined. In experiment 1, 65 sows were equally divided into the five following groups, between days 3 and 5 of the first or second estrous cycle post weaning: (1) controls (CTL); (2) 1 g of genistein fed daily (OR1); (3) 2 g of genistein fed daily (OR2); (4) two daily i.m. injections of 200 mg of genistein (IM400); and (5) two daily i.m. injections of 400 mg of genistein (IM800). Treatments were carried out for 10 days. In experiment 2, 10 sows were equally divided into two groups on day 90 of gestation, namely, controls (CTL) or 2 g of genistein fed daily for 10 days (OR2). In both trials, jugular blood samples were collected on days 1 (before treatment), 5 and 10 at 0730 h. In experiment 1, a blood sample was also collected at 1730 h on day 10 for CTL, IM400 and IM800 sows. In experiment 1, circulating concentrations of genistein on days 5 and 10 were greater in OR2, IM400 and IM800 than in CTL and OR1 group sows (P < 0.01). Daily dietary supplementation with 2 g of genistein resulted in blood concentrations that were similar to those in animals given daily two i.m. injections of 200 mg. Values of all isoflavones, except equol, which was not detectable, were greater in PM than in AM on day 10 (P < 0.01). In experiment 2, genistein concentrations were greater in OR2 compared with CTL on days 5 and 10 (P ⩽ 0.05). There was no difference in the genistein response to OR2 because of physiological status (i.e. weaned v. gestating, P > 0.1). Estradiol and IGF-I concentrations were not altered by any of the treatments (P > 0.1). Providing genistein either per os or via i.m. injections increased circulating concentrations of genistein in female swine within 5 days of the onset of treatment. The genistein response to i.m. injections of genistein was similar in weaned and late-pregnant sows, even though endogenous concentrations of estradiol differed. This response was specific in that estradiol, IGF-I and isoflavones other than genistein were not affected by treatments.
Dietary genistein stimulates mammary hyperplasia in gilts
- C. Farmer, M. F. Palin, G. S. Gilani, H. Weiler, M. Vignola, R. K. Choudhary, A. V. Capuco
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The possible role of the phytoestrogen genistein on prepubertal development of mammary glands, hormonal status and bone resorption was investigated in gilts. Forty-five gilts were fed a control diet containing soya (CTLS, n = 15), a control diet without soya (CTL0, n = 15) or the CTLS diet supplemented with 2.3 g of genistein daily (GEN, n = 15) from 90 days of age until slaughter (day 183 ± 1). Both basal diets were isonitrogenous and isocaloric. Jugular blood samples were obtained on days 89 and 176 to determine concentrations of isoflavone metabolites (on day 176 only), prolactin, estradiol, progesterone, insulin-like growth factor 1 (IGF1), and N-telopeptide of type I collagen (NTx; on day 176 only). At slaughter, mammary glands were excised, parenchymal and extraparenchymal tissues were dissected, and composition of parenchymal tissue (protein, fat, dry matter (DM), DNA) was determined. Histochemical analyses of mammary parenchyma were performed. Dietary genistein increased parenchymal protein (P < 0.05) while decreasing DM (P < 0.05) and tending to lower fat content compared with the CTLS, but not the CTL0, diet. There was more parenchymal DNA (1.26 v. 0.92 mg/g, P < 0.05) in GEN than CTLS gilts, likely reflecting an increase in the quantity of mammary epithelial cells. Circulating concentrations of genistein were increased in GEN gilts (P < 0.001) but concentrations of hormones or NTx (indicator of bone collagen resorption) were not affected by GEN (P > 0.1). Percentage of estradiol receptor alpha (ERα)-positive epithelial cells was lower (P < 0.05) in GEN than CTLS gilts, whereas 5-bromo-2′-deoxyuridine labeling index was unaltered (P > 0.1). Transcript levels for ERα, ERβ, IGF1, epidermal growth factor (EGF), epidermal growth factor receptor and transforming growth factor alpha were not altered by treatments. Supplementation of the diet with genistein during the growing phase in gilts, therefore, led to hyperplasia of mammary parenchymal tissue after puberty; yet, even though circulating genistein was increased, this was not accompanied by changes in mammary expression of selected genes or circulating hormone levels.