Responses of growth performance,antioxidant function,small intestinal morphology and mRNA expression of jejunal tight junction protein to dietary iron in yellow-feathered broilers

This study aimed to investigate the dose-effect of iron on growth performance,antioxidant function.intestinal morphology,and mRNA expression of jejunal tight junction protein in 1-to21-d-old yellow-feathered broilers.A total of 7201-d-old yellow-feathered maleb roilers were allocated to 9 treatments with 8 replicate cages of 10 birds per cage.The dietary treatments were consisted of a basal diet(contained 79.6 mg Fe kg^(-1))supplemented with 0,20,40,60,80,160,320,640,and 1,280 mg Fe kg^(-1)in the form of FeSO_(4)·7H_(2)O.Compared with the birds in the control group,birds supplemented with 20mg Fe kg^(-1)had higher average daily gain(ADG)(P<0.0001).Adding 640 and 1,280 mg Fe kg^(-1)significantly decreased ADG(P<0.0001)and average daily feed intake(ADFI)(P<0.0001)compared with supplementation of 20mg Fe kg^(-1).Malondialdehyde(MDA)concentration in plasma and duodenum increased linearly(P<0.0001),but MDA concentration in liver and jejunum increased linearly(P<0.05)or quadratically(P<0.05)with increased dietary Fe concentration.The villus height(VH)in duodenum and jejunum,and the ratio of villus height to crypt depth(V/C)in duodenum decreased linearly(P?0.05)as dietary Feincreased.As dietary Fe increased,the jejunal relative mRNA abundance of claudin-1 decreased linearly(P=0.001),but the jejunal relative mRNA abundance of zona occludens-1(ZO-1)and occludin decreased linearly(P?0.05)or quadratically(P?0.05).Compared with the supplementation of 20 mg Fe kg^(-1),the supplementation of640 mg Fe kg^(-1)or higher increased(P?0.05)MDA concentrations in plasma,duodenum,and jejunum,decreased VH in the duodenum and jejunum,and the addition of 1,280 mg Fe kg^(-1)reduced(P?0.05)the jejunal tight junction protein(claudin-1,ZO-1,occludin)mRNA abundance.In summary,640 mg of supplemental Fe kg^(-1)or greater was associated with decreased growth performance,increased oxidative stress,disrupted intestinal morphology,and reduced mRNA expression of jejunal tight junction protein.

In vitro effects of chitosan nanoparticles on proliferation of human gastric carcinoma cell line MGC803 cells

AIM： To investigate the effects of chitosan nanoparticles on proliferation of human gastric carcinoma cell line MGC803 in vitro and the possible mechanisms involved. METHODS： Chitosan nanoparticles were characterized by particle size, zeta potential, and morphology. After treatment with various concentrations of chitosan nanoparticles （25, 50, 75, 100 μg/mL） at various time intervals, cell proliferation, ultrastructural changes, DNA fragmentation, mitochondrial membrane potential （MMP）, cell cycle phase distribution and apoptotic peaks of MGC803 cells were analyzed by MTT assay, electron microscopy, DNA agarose gel electrophoresis, and flow cytometry. RESULTS： Chitosan nanoparticles exhibited a small particle size as 65 nm and a high surface charge as 52 mV. Chitosan nanoparticles markedly inhibited cell proliferation of MGC803 cells with an ICso value of 5.3 μg/mL 48 h after treatment. After treatment with chitosan nanoparticles, the typical necrotic cell morphology was observed by electron microscopy, a typical DNA degradation associated with necrosis was determined by DNA agarose electrophoresis. Flow cytometry showed the loss of MMP and occurrence of apoptosis in chitosan nanoparticles-treated cells. CONCLUSION： Chitosan nanoparticles effectively inhibit the proliferation of human gastric carcinoma cell line MGC803 in vitro through multiple mechanisms, and may be a beneficial agent against human carcinoma.

Expression levels of GSTA2 and APOD genes might be associated with carotenoid coloration in golden pheasant(Chrysolophus pictus) plumage

Carotenoids, which generate yellow, orange, and red colors, are crucial pigments in avian plumage. Investigations into genes associated with carotenoid- based coloration in avian species are important; however, such research is difficult because carotenoids cannot be synthetized in vertebrates as they are only derived from dietary sources. Here, the golden pheasant （Chrysolophus pictus） was used as a model in analysis of candidate gene expression profiles implicated in carotenoid binding and deposition. Using mass and Raman spectrometry to confirm the presence of carotenoids in golden pheasant feathers, we found C40H540 and C40H5602 in feathers with yellow to red colors, and in the rachis of iridescent feathers. The global gene expression profiles in golden pheasant skins were analyzed by RNA-seq and all six carotenoid binding candidate genes sequenced were studied by real- time PCR. STAR4, GSTA2, Scarbl, and APOD in feather follicles showed different expressions in red breast and orange nape feathers compared with that of iridescent mantle feathers. Further comparison of golden pheasant yellow rump and Lady Amherst＇s pheasant （Chrysolophus amherstiae） white nape feathers suggested that GSTA2 and APOD played a potential role in carotenoid-based coloration in golden pheasant.

Discovery of natural products capable of inducing porcine host defense peptide gene expression using cell-based high throughput screening

Background:In-feed antibiotics are being phased out in livestock production worldwide.Alternatives to antibiotics are urgently needed to maintain animal health and production performance.Host defense peptides(HDPs)are known for their broad-spectrum antimicrobial and immunomodulatory capabilities.Enhancing the synthesis of endogenous HDPs represents a promising antibiotic alternative strategy to disease control and prevention.Methods:To identify natural products with an ability to stimulate the synthesis of endogenous HDPs,we performed a high-throughput screening of 1261 natural products using a newly-established stable luciferase reporter cell line known as IPEC-J2/pBD3-luc.The ability of the hit compounds to induce HDP genes in porcine IPEC-J2 intestinal epithelial cells,3D4/31 macrophages,and jejunal explants were verified using RT-qPCR.Augmentation of the antibacterial activity of porcine 3D4/31 macrophages against a Gram-negative bacterium(enterotoxigenic E.coli)and a Gram-positive bacterium(Staphylococcus aureus)were further confirmed with four selected HDP-inducing compounds.Results:A total of 48 natural products with a minimum Z-score of 2.0 were identified after high-throughput screening,with 21 compounds giving at least 2-fold increase in luciferase activity in a follow-up dose-response experiment.Xanthohumol and deoxyshikonin were further found to be the most potent in inducing pBD3 mRNA expression,showing a minimum 10-fold increase in IPEC-J2,3D4/31 cells,and jejunal explants.Other compounds such as isorhapontigenin and calycosin also enhanced pBD3 mRNA expression by at least 10-fold in both IPEC-J2 cells and jejunal explants,but not 3D4/31 cells.In addition to pBD3,other porcine HDP genes such as pBD2,PG1-5,and pEP2C were induced to different magnitudes by xanthohumol,deoxyshikonin,isorhapontigenin,and calycosin,although clear gene-and cell type-specific patterns of regulation were observed.Desirably,these four compounds had a minimum effect on the expression of several representative inflammatory cytokine genes.Furthermore,when used at HDP-inducing concentrations,these compounds showed no obvious direct antibacterial activity,but significantly augmented the antibacterial activity of 3D4/31 macrophages(P<0.05)against both Gram-negative and Gram-positive bacteria.Conclusions:Our results indicate that these newly-identified natural HDP-inducing compounds have the potential to be developed as novel alternatives to antibiotics for prophylactic and therapeutic treatment of infectious diseases in livestock production.

Morphological Assessment of the Small Intestine of Broilers Fed Dietary Natural Zeolite Including Plant Extract

Many kinds of feed additives （e.g., probiotics, prebiotics, antibiotics, etc.） can be used in chicken diets to stimulate intestinal morphological maturation and, consequently, intestinal function. The aim of this trial was to investigate the possibility that natural zeolite including plant extract （ZEM） could encourage the hypertrophy of intestinal villi and the absorptive function of epithelial cells in broiler chickens. At 7 days old, 64 male Marshall Chunky broilers were divided into four groups, each with four replicates of four chickens. The birds were fed a basal mash diet supplemented with ZEM at 0% （control）, 0.05%, 0.1%, and 0.2% for 42 days. At 49 days old, four chickens from each treatment were killed to obtain intestinal samples. Morphological intestinal assessment was conducted using both a light and a scanning electron microscope. A tendency to increased villus height and cell area could be observed throughout the intestinal segments of the broilers fed ZEM. Particularly, the villus height in the duodenum was significantly increased （P 〈 0.05） in the 0.05% ZEM group when compared with the control group. The area of the villus cells tended to be higher in the ZEM groups in all intestinal regions and showed a significant increase （P 〈 0.05） in the ileal part of the 0.2% ZEM group. Also, the assessment of cells mitosis in the intestinal crypts showed significantly higher numbers （P 〈 0.05） in all levels of ZEM groups in the duodenum and a tendency to increase in value in the jejunum and ileum, as compared with the control group. More protuberated cells and multi-cell clusters were found in the ZEM groups than in the control group, resulting in a rough cellular surface. In conclusion, the present study might suggest that the intestinal villi and epithelial cells on the villus apical surface are hypertrophied in the ZEM groups, and that intestinal villi adapt to enhance their absorption capacity by increasing the absorptive surface area. These results are indicated that ZEM can provide effective nutritive value when used as a feed additive.

Potential dual expansion of domesticated donkeys revealed by worldwide analysis on mitochondrial sequences

Molecular studies on donkey mitochondrial sequences have clearly defined two distinct maternal lineages involved in domestication.However,domestication histories of these two lineages remain enigmatic.We therefore compared several population characteristics between these two lineages based on global sampling,which included 171 sequences obtained in this study(including Middle Asian,East Asian,and African samples)plus 536 published sequences(including European,Asian,and African samples).The two lineages were clearly separated from each other based on whole mitochondrial genomes and partial non-coding displacement loop(D-loop)sequences,respectively.The Clade I lineage experienced an increase in population size more than 8000 years ago and shows a complex haplotype network.In contrast,the population size of the Clade II lineage has remained relatively constant,with a simpler haplotype network.Although the distribution of the two lineages was almost equal across the Eurasian mainland,they still presented discernible but complex geographic bias in most parts of Africa,which are known as their domestication sites.Donkeys from sub-Saharan Africa tended to descend from the Clade I lineage,whereas the Clade II lineage was dominant along the East and North coasts of Africa.Furthermore,the migration routes inferred from diversity decay suggested different expansion across China between the two lineages.Altogether,these differences indicated non-simultaneous domestication of the two lineages,which was possibly influenced by the response of pastoralists to the desertification of the Sahara and by the social expansion and trade of ancient humans in Northeast Africa,respectively.

Fermentation Characteristics of Silage of Sugar Cane Treated with Calcium Oxide,Lactobacillus buchneri and Their Associations

We evaluated the fermentation products, the gaseous and effluent losses of sugarcane silages without calcium oxide (CaO) or with CaO addition, at the levels of zero, 0.8%, 1.6% and 2.4%, in association with the microbial additive Lactobacillus buchneri inoculated at the levels of zero, 50.000, 100.000 e 150.000 ufc/g of sugarcane, wet bases. The variety RB855536, harvested after 12 months of first growth was used. The experiment design was the completely randomized design, in a 4 × 4 factorial arrangement. They were evaluated in the silages, the contents of volatile fatty acids, lactic acid, ethanol, and the pH, as well as the gaseous and effluent losses. In the analysis of the data, the SAS system was utilized. It was observed on interaction effect of the chemical and microbial additive over the contents of lactic acid, acetic acid, propionic acid, butyric acid, ethanol and over the gaseous and effluents losses. However, there was no interaction effect regarding to pH. When it was observed on interaction effect of additives, the effects of the levels of one additive were evaluated by regression analysis in each level of each other, and vice-versa. The level 1.6% of CaO associated to the level 50,000 ufc/g of natural matter of Lactobacillus buchneri provided adequate levels of lactic acid (superior to 4.5%), and of acetic acid (around 1%), moderate content of propionic acid (0.55%), low content of butyric acid (0.05%) and controlled the production of ethanol and the gaseous and effluent losses. The pH of the silages were influenced by CaO addition, but were not affected consistently by microbial inoculation.

Relative Frequencies, Chemical Composition and in vitro Organic Matter Digestibility of Forage Consumed by Sheep in Humid Tropic of West Africa

The experiment was conducted during the short dry and the short rainy seasons in 2008. A botanical inventory and relative frequencies of forage selected by Djallonke sheep grazing were developed by observation and hand-plucking. Chemical composition and in vitro organic matter digestibility （IVOMD） of hand-plucked material was determined. From July to October, woody plants are a major component of the sheep diet. Sheep selected 40 woody plants and 21 species of herbaceous. During the dry season, grazing occurred more frequently in the fallows. The most frequent woody species belonged to Elaeis guineensis and Fagara zanthoxyloides. At the rainy season, a high proportion of Panicum maximum and Eleucine indica were sampled. Crude protein was high in woody leaves at the dry season. Crude fibre were more variable in herbaceous than in woody and herbaceous were generally higher in the levels of fibre. Chromolaena odorata, Citrus sinensis, Vitex doniana were very high in IVOMD. Spondias mombin, Moringa oleifera, Persea americana, Psidium guajava, Dialium guineense presented lowest values in IVOMD. Among herbaceous, Andropogon gayanus and Panicum maximum still presented a high value of IVOMD at the dry season. High proteins suggest woody with potential as nitrogen supplements to ruminants fed low quality forage during the dry season.

Parkia biglobosa (Mimosaceae) Leaves, Fruits’ Pulp, and Barks of Stem and Root Phytochemicals Contents and Their Antioxidant Activities

More and more Parkia biglobosa plants’ stem barks, and roots are collected for traditional medicine uses. This work aimed to quantify Parkia biglobosa plant parts total polyphenols, total flavonoids contents and their antioxidant activities. The hypothesis was that the plant parts would have different phytochemical contents. So, leaves, stem barks, root barks and mature fruit pulp were collected. Alongside the leaves, the stem and root barks were sliced, air-dried at room temperature for 30 days. Thereafter, the samples were crushed and sieved. Following this, the powders were extracted with distilled water by maceration and decoction. As a result, the decoction was more efficient than the maceration. Leaves had the highest total polyphenol content (14.68), followed by stem barks (11.69), and root barks (9.09 ± 0.43 mg GAE/g) (0.0001 ≤ p ≤ 0.0002). However, for total flavonoid contents, the stem barks were better than the other parts. Indeed, stem barks delivered 0.74 and were followed by leaves for 0.6, and root bark for 0.49 ± 0.02 mg EQ/g (0.0001 ≤ p ≤ 0.0169). Finally, the antioxidant activities were 3.07 and 2.86 ± 0.1 μmol T.E/g, respectively for stem bark, and leaves (p = 0.0532). In conclusion, there is no need to debark the stems, because, in decoction, leaves and stem barks would have the same efficacy.

Dietary copper supplementation modulates performance and lipid metabolism in meat goat kids

Forty-eight male Lezhi black goat kids with similar body weight((12.09±1.70)kg)and age((60±5)d)were used to determine the effect of dietary copper(Cu),in the form of reagent grade Cu sulfate(CuSO4∙5H2O),on performance,serum lipid profile,and the relative mRNA abundance of genes involved in lipid metabolism.Goat kids were stratified by body weight and randomly assigned to one of 4 treatment groups.Each treatment consisted of 12 replicate pens with each pen containing one goat kid.Treatment groups received the basal diet with no supplemental Cu(control),basal diet plus 10 mg of Cu kg^(-1)of dry matter(DM),basal diet plus 20 mg of Cu kg^(-1)of DM,or basal diet plus 30 mg of Cu kg^(-1)of DM.Goats were housed individually in pens and fed a high-concentrate pelleted diet for 60 d.Average daily gain,average daily feed intake and feed:gain of goats were not affected by dietary Cu supplementation(P>0.10).No differences were detected in serum total cholesterol,triglyceride,and high density lipoprotein cholesterol concentrations of goat kids fed with different Cu concentrations(P>0.05).However,serum low density lipoprotein cholesterol concentrations decreased linearly(P=0.01)as the concentration of dietary Cu increased.Intramuscular fat content of longissimus muscle increased(P=0.002)quadratically and liver Cu concentrations increased(P<0.001)linearly as dietary Cu concentration increased.Compared with the control,dietary supplementation of 20 mg Cu kg^(-1)DM decreased the relative mRNA abundance of fatty acid-binding protein 4(P=0.01)and lipoprotein lipase(P=0.05),and tended to decrease the relative mRNA abundance of carnitine palmitoyltransferase I(P=0.06)in longissimus muscle of goats.The relative mRNA abundance of peroxisome proliferator-activated receptor alpha(P<0.001),carnitine acetyltransferase(P=0.001),and carnitine palmitoyltransferase Ⅰ(P=0.001)were also decreased in liver by Cu supplementation.These results indicate that dietary supplementation of Cu modified lipid metabolism by increasing muscular fat and decreasing serum low density lipoprotein cholesterol,and the modification might be associated with the reduction of relative mRNA abundance of genes for oxidation of long-chain fatty acid in muscle and liver of Lezhi black goat kids.

Different characteristics of 40 kinds of free amino acids and 5-amino levulinic acid between wines and sakes(Japanese rice wines)

Based on a systemic analysis for the globa1 burden of disease study from 1990 to 2016,the Global Burden of Diseases(GBD)2016 Alcohol Collaborators reported that alcohol use was a 1eading nisk factor for globa1 disease burden and caused substantial health loss.However,alcoholic dnink(wines and sakes)contains abundant nutrients,particularly fermentable amino acids denived from grapes and rice.Amino acid nutrition and metabolism is strongly associated with health and preventiom against various diseases.The purpose of this study was to elucidate 40 kinds of free amino acid(FAA)profles in wines and sakes.In addition,the present study also provided the contents of 5-aminolevulinic acid(5-AI A)involved in the vital functionality of mitochondria.FAAs were detected and analyzed by use of an automatic amino acid analyzer.Aromatic amino acid,branched chain amino acid and Fischer ratio were calculated for each beverage.Individua1 FAAs showed significant differences between wines and sakes.Sakes abundantly contained almost all FAAs whereas wines had just a lttle content in some FAAs.Sakse were rich in arginine,glycine,alanine,valine,glutamic acid and leucine,while wines were remarkably nich in proline(Pro).Wines showed increased FAA levels in 6 kinds of variables(4-hydroxyproline,3-methy1 histidine,B-alanine,a-aminobutyric acid,Pro and a.-aminoadipic acid).In functionally characteristic FAAs,both wines and sakes were considerably abundant in y aminobutynic acid and 5-AI A The total amount of aromatic amino acid and branched chain amino acid were markedly higher in sakes than in wines.These results revealed that there were the apparent different characteristics in FAA profiles between wines and sakes.This difference resulted from fermentation process in wines(single fermentation)and sakes(paralle1 double fermentation).Forty kinds of FAA profiles are more usefu1 in investigating characteristics of the taste of wines and sakes.In future,detailed FAA analysis has the potential to find functional nutrients such as 5-AI A,whereas other foods generally contain ittle amount of these FAAs.

Acute exposure to high-peak-power pulsed microwaves affecting the histamine H_3 receptor expression in rat hippocampus

In the Morris Water Maze test, high-peak-power pulsed microwave (MW)-exposed rats displayed some learning and memory behavior dysfunctions, and their escape time and swimming distance to the submerged platform were longer than those of the sham-exposed rats. To understand the molecular mechanism involved, the reverse tran- scription-polymerase chain reaction (RT-PCR) and the Western-blotting technique were used for investigating the mRNA and protein expression patterns of the histamine H3 receptor (H3R) in rat hippocampus. High-peak-power pulsed microwave-exposure did not remarkably lead to the change in expression of H3R mRNA in rat hippocampi; however, it promoted the up-regulatory expression of the H3R protein, which was possibly triggered through the mi- togen-activated protein kinase (MAPK) pathways. Therefore, further investigation of the molecular mechanism of the MW effects on the learning and memory behaviors is required.

Growth Performance, Carcass Quality, Visceral Organs and Intestinal Histology in Broilers Fed Dietary Dried Fermented Ginger and/or Fermented Corncob Powder

To determine whether dried fermented ginger (DFG), fermented with Japanese mugwort silage juice, could be replaced by fermented corncob powder (FCP) as a of feed ingredient source without significant body weight decrease or damage to visceral organs (using gross anatomical observation), to intestinal villi (using light microscopy), or to the epithelial cells on the villus apical surface (using scanning electron microscopy) the following investigation was performed. Sixty-four male broilers were allotted to 4 groups: a basal diet group (control group), and basal diet groups with DFG at a level of 50 ppm;with DFG at 50 ppm and FCP at 250 ppm (50 ppm DFG + 250 ppm FCP group);and with FCP at a level of 500 ppm (500 ppm FCP group). Feed intake, body weight gain, feed efficiency, carcass quality, small intestinal length and weight, and visceral organ weight were not different among groups. Furthermore, regarding intestinal villus height, villus area and crypt depth, a significant difference was not found among the groups. When these values of the control were expressed as an index of 100, the duodenal villus height of the 50 ppm DFG + 250 ppm FCP group and the 500 ppm FCP group were 114 and 119, respectively. The duodenal villus area of the 50 ppm DFG + 250 ppm FCP group and the 500 ppm FCP group were 125 and 158, respectively. These villus heights and areas are thought to be activated. On the epithelial cells on the villus apical surface in the duodenum and jejunum, the 50 ppm DFG + 250 ppm FCP group had protuberated cells into the intestinal lumen and deeper cells at the sites of recently exfoliated cells, suggesting that these cells are activated. The present results indicate that small amounts of fermented corncob powder can be used as a feed supplement when mixed with fermented ginger powder, due to the synergy between the two ingredients, resulting in a 6% increase in body weight gain。

Growth performance,welfare traits and meat characteristics of broilers fed diets partly replaced with whole Tenebrio molitor larvae

The role of insects in animal nutrition has been reconsidered during recent years,paving the way for an increasing market for edible insects.Their protein and amino acid balance make them a promising source of protein for replacing high value proteins.Yellow mealworm,Tenebrio molitor L.(TM;Coleoptera:Tenebrionidae)larvae,have shown positive effects on broiler performance in several research studies and have a strong potential as a sustainable alternative protein source for monogastric animals.This study aimed to assess the effect of replacing various ratios of basal diets with T.molitor larvae on broiler performance as well as on several meat and welfare characteristics.For the study,120 one-day-old male chicks(Ross 308)were randomly allocated in 3 treatments and 4 replications(10 birds per pen).Birds of the control group(basal diet)were fed with typical commercial maize and soymeal-based rations in mash form.The other 2 groups were treated with the same diet,after replacing 5%and 10%with dried TM larvae,respectively.On d 35(end of trial),meat samples were collected and analysed.Body weight,feed intake,body weight gain and feed conversion ratio during the periods of 1 to 10 d(starter period),11 to 24 d(grower period),25 to 35 d(finisher period)and 1 to 35 d(total period)were assessed.Podo-dermatitis,diarrhoea,feather score and litter conditions were also assessed during the trial.The results indicated that TM larvae inclusion in the broilers'diet positively affected body weight gain values,as well as the carcass yield,the meat composition and various welfare traits.Additionally,the dietary treatments with TM larvae favourably affected meat composition and colour parameters,whereas there were also some positive effects on lipid and protein oxidation.Saturated fatty acids were decreased by the dietary supplementation whereas the polyunsaturated fatty acids to SFA ratio increased.In general,the study showed that whole TM larvae addition can provide a promising alternative to soybean meal in the diet of broilers,demonstrating a positive impact on growth,welfare and meat characteristics.