New biomarkers for increased intestinal permeability induced by dextran sodium sulphate and fasting in chickens

Increased intestinal permeability (IP) can lead to compromised health in chickens. As there is limited literature on in vivo biomarkers to assess increased IP in chickens, the objective of this study was to identify a reliable biomarker of IP using DSS ingestion and fasting models. Male Ross chickens (n = 48) were reared until day 14 on the floor pen in an animal care facility, randomized into the following groups: control, DSS and fasting (each with n = 16), and then placed in metabolism cages. DSS was administered in drinking water at 0.75% from days 16 to 21, while controls and fasted groups received water. All birds had free access to feed and water except the birds in the fasting group that were denied feed for 19.5 h on day 20. On day 21, all chickens were given two separate oral gavages comprising fluorescein isothiocyanate dextran (FITC‐d, 2.2 mg in 1 ml/bird) at time zero and lactulose, mannitol and rhamnose (LMR) sugars (0.25 g L, 0.05 g M and 0.05 g R in 2 ml/bird) at 60 min. Whole blood was collected from the brachial vein in a syringe 90 min post‐LMR sugar gavage. Serum FITC‐d and plasma LMR sugar concentrations were measured by spectrophotometry and high‐performance ion chromatography respectively. Plasma concentrations of intestinal fatty acid binding protein, diamine oxidase, tight junction protein (TJP), d ‐lactate and faecal α‐antitrypsin inhibitor concentration were also analysed by ELISA. FITC‐d increased significantly (p < 0.05) after fasting compared with control. L/M and L/R ratios for fasting and L/M ratio for DSS increased compared with control chickens (p < 0.05). TJP in plasma was significantly increased due to fasting but not DSS treatment, compared with controls. Other tests did not indicate changes in IP (p > 0.05). We concluded that FITC‐d and LMR sugar tests can be used in chickens to assess changes in IP.     

Gene expression and morphological changes in the intestinal mucosa associated with increased permeability induced by short‐term fasting in chickens

Short‐term fasting for 4.5 and 9 hr has been demonstrated to increase intestinal permeability (IP) in chickens. This study aimed to investigate the effects of 0, 4.5, 9 and 19.5 hr fasting on intestinal gene expression and villus‐crypt architecture of enterocytes in jejunal and ileal samples. On day 38, Ross‐308 male birds were fasted according to their group and then euthanised. Two separate intestinal sections (each 2 cm long, jejunum and ileum) were collected. One section was utilised for villus height and crypt depth measurements. The second section was snap‐frozen in liquid nitrogen for quantitative polymerase chain reaction (qPCR) analysis of tight junction proteins (TJP) including claudin‐1, claudin‐3, occludin, zonula occludens (ZO‐1, ZO‐2), junctional adhesion molecules (JAM) and E‐cadherin. Additionally genes involved in enterocyte protection including glucagon‐like peptide (GLP‐2), heat‐shock protein (HSP‐70), intestinal alkaline phosphatase (IAP), mammalian target of rapamycin (mTOR), toll‐like receptors (TLR‐4), mucin (MUC‐2), cluster differentiation (CD‐36) and fatty acid‐binding protein (FABP‐6) were also analysed. Normally distributed data were analysed using one‐way analysis of variance ANOVA. Other data were analysed by non‐parametric one‐way ANOVA. Villus height and crypt depth were increased (p < .05) only in the ileum after fasting for 4.5 and 9 hr compared with non‐fasting group. mRNA expression of claudin‐3 was significantly reduced in the ileum of birds fasted for 9 and 19.5 hr, suggesting a role in IP modulation. However, all other TJP genes examined were not statistically different from control. Nevertheless, ileal FABP‐6 of all fasted groups was significantly reduced, which could possibly be due to reduced bile acid production during fasting.     

Effects of sinomenine in LPS‐associated diseases are related to inhibition of LBP,Mac‐1, and L‐selectin levels

The aim of the research was to investigate the anti‐endotoxin and anti‐inflammatory effects of Sinomenine, an agent commonly found in Chinese herbal medicines. Endotoxin (i.e., 1 mg lipopolysaccharide (LPS)/kg)) was administered via intraperitoneal (IP) injection to piglets in high‐, middle‐, and low‐dose sinomenine groups. Piglets were then treated with 1, 5 or 10 mg/kg sinomenine, intramuscularly (i.m.), 3 hr after LPS. Vehicle was administered, as above, to drug control group piglets followed 3 hr later by 10 mg/kg sinomenine i.m.. LPS control group piglets were challenged with 1 mg/kg LPS IP, followed by vehicle i.m., and naïve control piglets were treated with normal saline IP, followed by normal saline i.m., as above. Temperatures were measured, and blood samples were collected from the precaval veins of piglets at 12, 24, and 48 hr post‐LPS or vehicle injection. Clinical signs were recorded, and index levels were analyzed via ELISA. Sinomenine was found to reduce the incidence and severity of LPS‐induced toxicities, including body temperature elevation, cell adhesion, and systemic inflammation. These data suggest that sinomenine may be effective for regulating inflammatory responses and has the potential for use as an anti‐endotoxin therapy.     

Relieving effect of Artemisia argyi aqueous extract on immune stress in broilers

This experiment aimed to investigate the relieving action of Artemisia argyi aqueous extract (AAE) on immune stress in broiler chickens. A 2 × 2 factorial design was used to test the effect of 2 dietary treatments (adding 0 or 1000 mg/kg AAE) and 2 immune stress treatments (injecting saline or lipopolysaccharide (LPS)). A total of 96 one‐day‐old Arbor Acres (AA) broilers were randomly divided into four treatment groups with six replicates, four birds in each replicate. Broilers in Treatment groups 1 and 2 were fed with the basal diet, and those in Treatment groups 3 and 4 were fed with the experimental diet supplemented with 1000 mg/kg AAE. On days 14, 16, 18 and 20, broilers in both Treatments 1 and 3 were injected intra‐abdominally with LPS solution at the dose of 500 μg LPS per kg BW with the LPS dissolved in sterile saline at a concentration of 100 μg/ml, and those in Treatments 2 and 4 were injected intra‐abdominally with equal amount of sterile 0.9% saline. Blood samples were collected on days 21 and 28. The results showed that dietary supplementation of AAE prevented reductions in average daily gain (ADG) and average daily feed intake (ADFI) of broilers caused by LPS on d 15–21. On day 21, the injection of LPS increased serum adrenocorticotropic hormone (ACTH) and corticosterone (CORT); meanwhile, feeding AAE reduced the rise of CORT caused by LPS. Immune parameters such as interleukin‐1 (IL‐1), interleukin‐2 (IL‐2), interleukin‐6 (IL‐6), immunoglobulin G (IgG) and immunoglobulin A (IgA) were also improved by LPS, but the content of IL‐2 and IgG in broilers fed with AAE diet was significantly lower than that of broilers fed with control diet. All the data suggested that diets supplemented with AAE could relieve the immune stress response of broilers.     

Influence of procyanidin supplementation on the immune responses of broilers challenged with lipopolysaccharide

In the present study, the effect of dietary procyanidin (PCA, from pine needles) supplementation on the innate immunity of broilers were investigated. The experiment was designed as a 2 × 4 factorial arrangement (eight cages / treatment; six birds (one‐day‐old) / cage) with dietary PCA concentrations (0, 0.05, 0.075 and 0.1%) and two immune treatments (injection of lipopolysaccharide (LPS) (0.5 mg/kg body weight) or saline). LPS was dissolved in sterile 9 g/L (w/v) NaCl solution at 16, 18, 20 days of age to mimic immune stress. The remaining birds were injected with saline as a placebo. The results indicated that, prior to LPS challenge, the PCA diet had no significant effect on bird growth performance. The injection of LPS was also not associated with any significant changes in poultry performance. LPS injection increased the activity of nitrogen oxides (NOx) and the concentrations of inflammatory cytokines (interferon‐γ (IFN‐γ), interleukin‐1β (IL‐1β), IL‐2, IL‐4, IL‐6 and IL‐10) in serum; dietary PCA decreased these concentrations (P < 0.05) in the PCA 0.1% group, further illustrating the immune effect of PCA. In conclusion, PCA supplementation has a beneficial effect on LPS challenge, which may be associated with the inhibition of the secretion of cytokines and decrease in the proinflammatory marker NOx.     

Effect of humic acids on intestinal viscosity,leaky gut and ammonia excretion in a 24 hr feed restriction model to induce intestinal permeability in broiler chickens

The purpose of this study was to evaluate the effect of humic acids (HA) on intestinal viscosity, leaky gut and ammonia excretion in a 24 hr feed restriction (FR) model to induce intestinal permeability in chickens. One‐day‐old male Cobb‐Vantress broilers were randomly allocated to one of two groups (n = 25 chickens), with or without 0.2% of isolated HA from worm‐compost, and placed in brooder batteries. Chicks had ad libitum access to water and feed for 14 days. Intestinal permeability was induced by 24 hr FR starting at 14 days. At 15 days of age, chickens in both groups were given an appropriate dose of fluorescein isothiocyanate dextran (FITC‐d) by oral gavage. Intestine and liver samples were also collected to evaluate viscosity and bacterial translocation (BT), respectively. An increase (p < .05) in intestinal viscosity was observed in the experimental group consuming 0.2% of HA and was confirmed in a published in vitro digestion model that simulates the chemical and physical conditions of the crop, proventriculus and intestine of chickens. Furthermore, the treated group also showed a significant reduction in FITC‐d, liver BT and ammonia in the manure. These results suggest that HA have a positive impact in intestinal integrity in chickens.     

Physiological adaptive indicators in fasted neonate broiler chicks in response to calcium gluconate injection

Four hundred and eighty mixed‐sex broiler chicks aged 3 h after hatching were allotted according to a completely random design in a 6 × 2 × 2 factorial schedule into two groups of 12 replications of 20 chicks each. The main experimental factors were fasting for 0, 6, 12, 24, 36 and 48 h after chick placement and calcium gluconate (Ca‐glu) injection (0 and 0.6 ml). Live body weight (BW) of chicks decreased linearly (Y = 43.36–0.109BW_0 h, r² = 0.876) as neonatal fasting extended. Injection of 0.6 ml Ca‐glu at 3 h post‐hatching did not affect weight loss of chicks. Yolk residuals (YR) utilized linearly (Y = 5.75–0.062YR, r² = 0.956) by 0.062 g/h in neonate fasted chicks up to 48 h, showing no effect of Ca‐glu injection. Neonatal fasting periods longer than 12 h increased liver weight (p < 0.05). The mean absolute and proportional (% of BW_0 h) breast and leg weight were reduced linearly as neonatal fasting extended (p < 0.05). Serum glucose concentration increased up to 6 h and then reduced linearly to 150 mg/dl after 48‐h fasting. The Ca‐glu treatment influenced serum glucose level for a short period up to 6 h of fasting. Serum Ca concentration sharply increased up to threefolds in the birds received Ca‐glu injection resulting in acute hypercalcemia, then decreased to the initial level after 24‐h feed withdrawal (p < 0.05). The mean serum level for creatinine, uric acid, cholesterol, HDL, albumins and total proteins significantly increased during the fasting periods of 6 to 48 h and significantly elevated in the birds receiving 0.6‐ml Ca‐glu injection compared with the non‐treated chicks (p < 0.05). It was concluded that subcutaneous administration of 0.6 ml Ca‐glu in the chick's neck did not suitably support the increased metabolic demands for glucose and calcium in feed‐deprived neonate chicks.     

Evaluating two multistrain probiotics on growth performance,intestinal morphology,lipid oxidation and ileal microflora in chickens

An experiment was conducted to investigate the supplementation of two commercially available multistrain probiotics as an alternative to antibiotics on growth performance, intestinal morphology, lipid oxidation and ileal microflora in broiler chickens. A total of 280‐day‐old ROSS 308 mixed‐sex broiler chickens with an average initial body weight of 42 ± 0.5 g were randomly divided into four treatments with five replicate cages of 14 birds each cage in a completely randomized design and fed with the following diets for 42 day: (a) control (CON) (antibiotic‐free diet), (b) antibiotic (ANT) (CON + Avilamycin 150 g/ton feed), (c) probiotic A (CON + Protexin^® 150 g/ton feed) and (d) probiotic B (CON + Bio‐Poul^® 200 g/ton feed). The results showed the broilers fed the ANT diet had greater average daily gain than broilers fed the CON diet during day 1–14 (p < 0.05). At day 42, two birds were randomly selected per replicate for evaluation intestinal morphology, lipid oxidation and ileal microflora. birds fed diet supplemented with probiotic A and probiotic B increased villus height and goblet cells numbers in the jejunum and villus height to crypt depth ratio and villus height in the ileum as compared to birds fed CON diet (p < 0.05). The malondialdehyde value was reduced (p < 0.05) in the ANT, probiotic B and probiotic A groups compared with the CON group. The Lactobacillus population was increased and Clostridium spp. population decreased in the ileum of broilers fed diets containing the probiotic B and probiotic A compared with those fed CON diet (p < 0.05). The results from this study indicate that the probiotic A (Protexin^®) and probiotic B (Bio‐Poul^®) used in this trial may serve as alternatives to ANT.     

Improving effect of dietary soybean phospholipids supplement on hepatic and serum indexes relevant to fatty liver hemorrhagic syndrome in laying hens

In order to investigate the effect of dietary soybean phospholipid supplement on hepatic and serum indexes relevant to fatty liver hemorrhagic syndrome (FLHS) in layers, 135 300‐day‐old Hyline Brown layers were randomly divided into three groups (control, pathology and prevention), and each group had 45 layers with three replicates. Birds in the three groups were respectively fed the control diet, high‐energy low‐protein diet and high‐energy high‐protein diet affixed with 3% soybean phospholipid instead of maize. Results showed in the 30th day, birds’ livers in the pathology group became yellowish, enlarged in size and had hemorrhagic spots, while the prevention and control groups’ layers did not have such pathological changes. Contents of triglyceride, total cholesterol, low‐density lipoprotein – cholesterol, non‐esterified fatty acid and malondialdehyde in serum or liver homogenate in prevention and control groups were remarkably lower than those in the pathology group (P < 0.05 or P < 0.01), as with the activities of glutamic oxalacetic transaminase and glutamic‐pyruvic transaminase (P < 0.01); high‐density lipoprotein – cholesterol value was strikingly higher than that of the pathology group (P < 0.01). It is suggested dietary soybean phospholipids supplement may effectively improve hepatic and blood indexes relevant to FLHS, which provides a new point for preventing FLHS occurrence rate in laying flocks and treating human non‐alcohol fatty liver disease.     

Effects of berberine on the growth performance,antioxidative capacity and immune response to lipopolysaccharide challenge in broilers

This study evaluated the effects of berberine on growth performance, immunity, haematological parameters, antioxidant capacity, and the expression of immune response‐related genes in lipopolysaccharide (LPS)‐challenged broilers. We assigned 120 one‐day‐old male broilers (Ross 308) to two treatment groups; each group included two subgroups, each of which included six replicates of five birds per replicate. The experiment used a 2 × 2 factorial arrangement with berberine treatment (0 or 60 mg/kg dietary) and challenge status [injection of saline (9 g/L w/v) or LPS (1.5 mg/kg body weight)] as the main factors. On days 14, 16, 18 and 20, broilers were intraperitoneally injected with LPS or physiological saline. Blood and liver samples were collected on day 21. Dietary berberine supplementation significantly alleviated the compromised average daily gain and average daily feed intake (p < 0.05) caused by LPS. The LPS challenge led to increased lymphocyte and white blood cell (WBC) counts, malondialdehyde (serum and liver) content, and immunoglobulin G and M, tumour necrosis factor‐α (TNF‐α) and interleukin‐1β (IL‐1β) expression (p < 0.05) and significantly reduced serum total superoxide dismutase (T‐SOD) activity (p < 0.05). Dietary berberine significantly mitigated the LPS‐induced decreases in the mRNA expression of nuclear factor‐kappa B (NF‐κB), TNF‐α, IL‐1β, inducible nitrite synthase and cyclooxygenase‐2 (p < 0.05) in the liver. In conclusion, berberine supplementation has a positive effect on LPS challenge, which may be related to the increase in antioxidant enzyme activity and inhibition of both NF‐κB signalling and the expression of inflammatory mediators.     

Effects of dietary supplemental methionine source and betaine replacement on the growth performance and activity of mitochondrial respiratory chain enzymes in normal and heat‐stressed broiler chickens

This study aimed to evaluate the effects of dietary supplemental methionine (Met) source and betaine (Bet) replacement for Met on performance and activity of mitochondrial respiratory chain enzymes (MRCEs) in normal and heat‐stressed broiler chickens. Total of 1,200‐day‐old Ross 308 chicks were allocated to two houses, each consisted of 12 treatments, five replicates of 10 birds each with 2 × 2×3 × 2 (temperature × Met source × Met level × Bet, respectively) split‐plot factorial arrangement. Met level in the basal diets was 70% requirements (Req) that was increased to the requirement or 130% by supplemental dl ‐ or l ‐Met. Bet was or was not substituted at the rate of 30% supplemental dl ‐ or l ‐Met. Feed conversion ratio (FCR) in chicks fed 70% l ‐Met was lower than those fed 70% dl ‐Met diet during 1–10 days (p = 0.04). Broilers fed diets containing requirement or 130% Met, regardless of its source, showed higher weight gain (WG) than those received 70% Met diet during 11–42 days (p < 0.001). Feed intake (FI) of broilers fed 130% Met diet was decreased compared to other two groups during 11–42 days (p < 0.05). One hundred thirty percent Met requirement diet resulted in lower FCR comparing to other two groups during 11–42 days (p < 0.001). Heat‐stressed birds grew less than those under normal condition (p < 0.05). Broilers fed Req Met diet under normal temperature exhibited higher activities of complexes (Cox) I and III (p < 0.05). Cox I activity in heat‐stressed birds fed Bet + diet was similar to those fed Bet‐diet under normal temperature (p = 0.046). It is concluded that performance and the activities of Cox I and III were increased as the level of Met increased. Bet replacement for 30% supplemental Met resulted in similar consequences comparing to non‐Bet replacement diets on performance, but increased the activity of Cox III. l ‐Met was effective than dl ‐Met at the cellular level. High ambient temperature depressed performance and MRCE activity.     

Bacillus subtilis B21 and Bacillus licheniformis B26 improve intestinal health and performance of broiler chickens with Clostridium perfringens‐induced necrotic enteritis

This study investigated the influence of Bacillus‐based probiotics on performance and intestinal health in broiler challenged with Clostridium perfringens‐induced necrotic enteritis. One‐day‐old Arbor Acre (n = 480) were randomly assigned to four treatments with 10 cages of 12 birds: (a) basal diet negative control (NC), with no probiotics nor antibiotics formulated to contain 2,930 and 3,060 kcal/kg with 24.07 and 15.98% CP, for starter and finisher diet, respectively, (b) basal diet + enramycin (5 mg/kg), an antibiotic growth promoter (AGP); (c) basal diet + Bacillus subtilis B21 at 2 × 10⁹ CFU per g (BS); (d) basal diet + Bacillus licheniformis B26 at 2 × 10⁹ CFU per g (BL); growth performance, intestinal morphology, intestinal lesion scores, short‐chain fatty acids (SCFAs) and mucosal barrier tight junction's (TJ) mRNA expression were assessed. NC‐ and BL‐fed groups showed higher (p = 0.005) average daily feed intake from d1 to d21 than AGP and BS, whereas BS‐ and AGP‐fed groups showed higher average daily weight gain from d22 to d42 and d1 to d42 of age. Higher mortality rate of (12.5%) and lower of (5.5%) were recorded in AGP and NC fed‐groups respectively, lesion score was higher in BS and BL than in AGP, while no lesion was observed in NC group, results revealed higher duodenum and jejunum villus height to crypt depth (VH:CD) compared with NC and BS. Probiotics‐fed groups showed higher total (SCFAs), acetic and butyric acid concentrations at d21 post‐challenge (PC) than other groups. The expression of claudin‐1 was upregulated in duodenum (d7) PC and in jejunum (d7) and (d21) PC in BL group, while at d21 PC, the expression of occludens was higher in jejunum and ileum by AGP and BL. The present study indicated both BS and BL have some similarity with AGP in preventing or partially preventing NE effect in broilers.     

Effects of dietary phytoncides extracted from Korean pine (Pinus koraiensis) cone on performance,egg quality,gut microflora,and immune response in laying hens

This study was conducted to evaluate the effects of dietary phytoncides extracted from discarded Korean pine cones (Pinus koraiensis) on the performance, egg quality, immune response and gut microflora in laying hens. A total of 400 Hy‐Line brown laying hens (50‐week old) were allotted into four dietary treatments including a control diet or a diet supplemented with phytoncides at 0.002%, 0.004% and 0.008%. During the 6 weeks of experimental feeding, 0.008% of dietary phytoncides improved egg production, feed conversion ratio (p < 0.05), but not feed intake, egg weight or feed efficiency. Although dietary phytoncides had no effect on egg quality, decreases in Haugh units depending on storage periods were improved by 0.008% of dietary phytoncides (p < 0.05). To investigate the roles of dietary phytoncides on the alteration of the immune response during inflammation, lipopolysaccharide (LPS) or saline was intraperitoneally injected into 10 hens per diet group on the end date of the experimental feeding period. Serum immunoglobulins and splenic cytokine expression at mRNA levels were then measured at 4 hr postinjection. Although the levels of IgA were decreased by LPS injection in all dietary groups, dietary phytoncides at 0.008% showed a higher level of IgA by LPS (p < 0.05). Interestingly, although LPS injection resulted in an enhanced expression of proinflammatory cytokines such as IL‐1β and IL‐6, dietary phytoncides at 0.008% showed less increased levels of them (p < 0.05). Gut microflora was examined from 10 hens per diet group at the end of the experimental period. While the number of Lactobacillus spp. was increased (p < 0.05), Escherichia coli counts in the cecal contents were decreased by 0.008% of dietary phytoncides. Taken together, these results demonstrate that dietary supplementation of 0.008% phytoncides improved the egg production, immune responses during inflammation and gut microflora in laying hens.     

Deoxynivalenol induces oxidative stress,inflammatory response and apoptosis in bovine mammary epithelial cells

Deoxynivalenol (DON) is a toxic secondary metabolite produced by Fusarium graminearum. It is one of the most common feed contaminants that poses a serious threat to the health and performance of dairy cows. This study investigated the in vitro cytotoxicity of DON on bovine mammary epithelial cells (MAC‐T). DON at different concentrations (0.25, 0.3, 0.5, 0.8, 1 or 2 μg/ml) inhibited the growth of MAC‐T cells after 24 hr of exposure (p < .001). DON at 0.25 μg/ml increased lactate dehydrogenase (LDH) leakage (p < .05); decreased glutathione (GSH) levels (p < .001), total superoxide dismutase (T‐SOD) activity and total antioxidant capacity (T‐AOC; p < .01); and increased malondialdehyde (MDA) concentration (p < .01) in MAC‐T cells after 24 hr of exposure. We also observed that DON increased reactive oxygen species (ROS) levels in cells incubated for 9, 15 and 24 hr (p < .001). DON at 0.25 μg/ml triggered oxidative damage in MAC‐T cells. Furthermore, it induced an inflammatory response in the cells incubated for 9, 15 and 24 hr (p < .05) by increasing the mRNA expression levels of nuclear factor kappa B, myeloid differentiation factor 88 (MyD88), tumour necrosis factor‐α (TNF‐α), interleukin‐1β (IL‐1β), IL‐6, cyclooxygenase‐2 and IL‐8. We further examined the effect of DON on apoptosis. DON prevented normal proliferation of MAC‐T cells by blocked cell cycle progression in 24 hr (p < .001). In addition, the apoptosis rate measured using annexin V‐FITC significantly increased (p < .05) with increase in the mRNA expression level of Bax (p < .01) and increase in the Bax/Bcl‐2 ratio (p < .01) in cells incubated for 24 hr. In summary, DON exerts toxic effects in MAC‐T cells by causing oxidative stress, inducing an inflammatory response, affecting cell cycle and leading to apoptosis.     

Impact of restricting feed and probiotic supplementation on growth performance,mortality and carcass traits of meat‐type quails

The objective of this study was to evaluate the effects of quantitative feed restriction, along with dietary supplementation with a probiotic blend (Protexin) as a natural growth promoter, on the performance, water consumption, mortality rate and carcass traits of meat‐type quails. A total of 250 1‐day unsexed quails were randomly allocated to five equal groups in a completely randomized design. The first group (A) fed a basal diet without any restriction (24 hr/day); the second group (B1) fed the basal diet for 20 hr/day; the third group (B2) fed the basal diet enriched with probiotic (0.1 g/kg diet) for 20 hr/day; the fourth group (C1) fed the basal diet for 16 hr/day; and the fifth group (C2) fed the basal diet enriched with probiotic (0.1 g/kg diet) for 16 hr/day. Birds were fed ad‐libitum from 0–14 days of age, and then the feed restriction regimes started from 14 till 28 days of age. Results showed that quails in the control‐group consumed more feed and water than the other treatment groups (p < .01), however their body weights did not differ (p > .05) compared with the other treated groups. The best feed conversion values were achieved in quails supplemented with probiotic blend (B2 and C2) in comparison with the other groups (p < .01). Feeding probiotic had a positive effect on bird health which reduced the mortality rate. Further, mortality rate was significantly reduced (p < .05) by feed restriction, with or without probiotic supplementation. No carcass parameters were significantly affected (p > .05) by treatments. Our results show that quail could be reared under a feed restriction system, for 4–8 hr daily, along with dietary supplementation of probiotic as growth promoter for better growth performance.     

Effect of exogenous progesterone administration on luteal sensitivity to PGF during the early development of the corpus luteum in mares and cows

The objective of this study was to determine the effect of exogenous progesterone administration at ovulation and during the early development of the CL, on its future sensitivity to a single administration of PGF2a in mares and cows. Horse Retrospective reproductive data from an equine clinic in the UK during three breeding seasons were used. Mares were divided into: control group, cycles with single ovulations; double ovulation group cycles with asynchronous double ovulations; and PRID group: cycles with single ovulations and treatment with intravaginal progesterone device (CIDR) immediately after the ovulation. All mares were treated with d‐cloprostenol (PGF) at either: (i) 88 hr; (ii) 96 hr; (iii) 104 hr; or (iv) 112 hr after the last ovulation. Cattle A total of nine non‐lactating Holstein cows were used. All cows were administered PGF14 d apart and allocated to one of two groups control group GnRH was administered 56 hr after the second PGF administration. CIDR group CIDR was inserted at the same time of GnRH administration. All cows were administered PGF at 120 hr post‐ovulation. The complete luteolysis rate of mares with double ovulation (66.7%) and those treated with exogenous progesterone (68.4%) was significantly higher than the rate of mares with single ovulation (35.6%) at 104 hr. In the cow, however, the treatment with CIDR did not increase the luteolytic response in cows treated at 120 hr post‐ovulation. In conclusion, the degree of complete luteolysis can be influenced by increasing the concentration of progesterone during the early luteal development in mares.     

Dietary conjugated linoleic acids alleviate early inflammatory response caused by lipopolysaccharide injection in male broiler chicks

An experiment was undertaken to determine if dietary conjugated linoleic acid (CLA) influences the early stage of the inflammatory response caused by a single injection of Salmonella enteritidisi lipopolysaccharide (LPS) in male broiler chicks. Chicks (7 days old) were fed either 0 or 10 g CLA/kg in their diet for 21 days. At 28 days old, birds were injected intraperitoneally with 1.5 mg LPS/kg body weight after 12 h fasting. Changes in feed intake, rectal temperature and plasma α‐1 acid glycoprotein and ceruloplasmin concentrations were measured during 24 h after LPS injection. Chicks fed the CLA diet were fed more feed for 24 h, had a lower rectal temperature at 9 and 24 h and a lower plasma α‐1 acid glycoprotein concentration at 24 h after LPS injection than chicks fed the basal diet. Plasma ceruloplasmin concentration tended to be lower in chicks fed the CLA diet than in chicks fed the basal diet. The results suggest that dietary CLA alleviate undesirable early inflammatory response due to LPS injection in male broiler chicks.     

The effect of different high‐fat diets on erythrocyte osmotic fragility,growth performance and serum lipid concentrations in male,Japanese quail (Coturnix coturnix japonica)

Poultry diets are formulated with additional animal fat or vegetable oils to improve growth rate and feed conversion efficiency. High‐fat diet feeding in rats and fish has been shown to result in alterations in the phospholipid composition and cholesterol content of the erythrocyte membrane, in turn affecting erythrocyte osmotic fragility. In contrast, the few studies performed using high‐fat diet feeding in avian species show no changes in erythrocyte osmotic fragility. This study made use of the Japanese quail as no data exists on investigation of this species with respect to high‐fat diet feeding and erythrocyte osmotic fragility. Fifty‐seven male quail were randomly divided into six groups and fed either a standard diet (commercial poultry feed) or one of five high‐fat diets (commercial poultry feed with 22% of either coconut oil, lard, palm oil, soya bean oil or sunflower oil on a weight/weight basis) for 12 weeks. All birds on the high‐fat diets were significantly heavier (p < 0.05) after the 12‐week feeding period, than when commencing the dietary intervention. Serum triglyceride concentrations of birds in all high‐fat diet groups were significantly lower (p < 0.05) than birds in the standard diet group, whereas only birds in the palm oil group had significantly lower (p < 0.05) serum cholesterol concentrations compared to the standard diet group. Fragiligrams of erythrocytes from birds in the various dietary groups were similar. High‐fat diet feeding with different types of additional fat did not affect the osmotic fragility of the quail erythrocytes. Feeding quail high‐energy diets of varying degrees of fatty acid saturation was well tolerated and did not seem to affect the overall health status of the birds. Resistance of avian erythrocytes to modification by excess dietary fat may be a general characteristic of avian erythrocytes.     

β‐carotene attenuates lipopolysaccharide‐induced inflammation via inhibition of the NF‐κB,JAK2/STAT3 and JNK/p38 MAPK signaling pathways in macrophages

β‐carotene is one of the most abundant carotenoids, has potential anti‐inflammatory effect, it has been reported that β‐carotene could suppress LPS‐induced inflammatory responses by inhibiting nuclear factor kappa B (NF‐κB) translocation, but the more detailed molecular mechanisms underlying the anti‐inflammatory action of β‐carotene remain to be fully understood. In this study, we investigated the influence of β‐carotene on the activation of JAK2/STAT3, MAPK, and NF‐κB signaling pathway induced by LPS in RAW264.7 cells and peritoneal macrophages. Cells were treated with different concentrations of β‐carotene for 3 hr after LPS treatment for 24 hr. The mRNA expression and the release of IL‐1β, IL‐6, and TNF‐α were evaluated by RT‐PCR and ELISA, and the level of signaling proteins of JAK2/STAT3, MAPK, and NF‐κB signaling pathway were detected by Western blot. The results showed that β‐carotene significantly suppressed (p < 0.05) LPS‐induced release of IL‐1β, IL‐6, and TNF‐α and their mRNA expression. LPS‐induced JAK2/STAT3, IκB/NF‐κB p65, JNK/p38 MAPK signal activation were significantly attenuated (p < 0.05) by β‐carotene in a dose‐dependent manner. In conclusion, β‐carotene could attenuate LPS‐induced inflammation via inhibition of the NF‐κB, JAK2/STAT3, and JNK/p38 MAPK signaling pathways in macrophages.     

Influence of feeding a fish oil‐containing diet to mature,overweight dogs: Effects on lipid metabolites,postprandial glycaemia and body weight

The objective of this study was to determine the effect of feeding a fish oil (FO)‐containing diet on lipid and protein metabolism, postprandial glycaemia and body weight (BW) of mature, overweight dogs. Seven female dogs were randomly assigned to one of two isonitrogenous and isocaloric diets, control (CO) or FO (FO), in a crossover design. Experimental periods were 69 day, separated by a washout period of 30 day. At the beginning of the experiment, and at 30 and 60 day of feeding the experimental diets, the dogs were infused with D‐glucose (2 g/kg BW) through an intravenous catheter. Blood samples were collected for 3 hr to perform a glucose tolerance test. Nitrogen balance measurements began at 06:30 on d 63 of each experimental period and ended at 06:30 on d 69. On d 66 of each period, a single dose (7.5 mg/kg) of ¹⁵N‐glycine was administered orally for determination of protein turnover. Incremental area under the curve and glucose concentration at peak did not differ between treatments or among sampling days within treatment. Glucose half‐life tended to decrease (p < .10) in the FO treatment on day 30 when compared to baseline (day 0). β‐hydroxybutyrate, non‐esterified fatty acid (NEFA) and triglycerides did not differ within or between treatments. Cholesterol decreased (p < .05) on the FO treatment on day 30, 60 and 69 when compared to day 0. High‐density lipoprotein (HDL) decreased (p < .05) in the FO treatment on day 69 when compared to day 0. Body weight, food intake, faecal excretion, DM and N digestibilities, N balance and protein turnover were not different between diets. Overall, FO‐containing diet decreases cholesterol in mature overweight dogs; however, further research is warranted to verify the effects of FO on glucose metabolism.