Jejunum Of Chickens Research Articles

Effects of dietary Allium hookeri root on growth performance and antioxidant activity in young broiler chickens

The objective of this experiment was to evaluate the antioxidant potential of Allium hookeri (AH) root in two forms, powdered AH root and fermented powdered AH root, to demonstrate its value as an antibiotic alternative feed additive for broiler chickens. A total of 125 male Ross-708 broiler chickens were randomly assigned to five groups (n = 25 birds/group) and fed standard diets supplemented with root or fermented root of AH at two different levels (1% or 5%). Control birds were provided with non-supplemented basal diets. Body weights was measured at days 14 and 21 of age. To monitor antioxidant activities, heme oxygenase (HMOX), aflatoxin B1 aldehyde reductase (AFAR), superoxide dismutase 1 (SOD1), and catalase (CAT) enzyme levels were quantified by real-time PCR in the jejunums 21-day-old birds. Also, serum levels of SOD, CAT, and malondialdehyde (MDA) were measured using commercial kits. The results showed greater body weight gains at day 14 in chickens fed diets supplemented with 1% AH root, as compared to the control group (p < 0.05). Up-regulated transcript levels of AFAR, HMOX1, and CAT were observed in the jejunum of chickens fed diets supplemented with AH. The serum levels SOD and CAT were significantly increased (p < 0.05) in groups treated with AH, whereas MDA levels were decreased in groups fed diets supplemented with AH, as compared to the control group. These results indicated that an optimum level of dietary AH supplementation to young broiler chickens influences growth and improves antioxidant activities.

Supplementation of Vitamin E Protects Chickens from Newcastle Disease Virus-Mediated Exacerbation of Intestinal Oxidative Stress and Tissue Damage

Background/Aims: Newcastle disease virus (NDV) causes a highly devastating and contagious disease in poultry, which is mainly attributed to extensive tissue damages in the digestive, respiratory and nervous systems. However, nature and dynamics of NDV-induced oxidative stresses in the intestine of chickens remain elusive. Methods: In this study, we examined the magnitude of intestinal oxidative stress and histopathological changes caused by the virulent NDV infection, and explored the protective roles of vitamin E (vit. E) in ameliorating these pathological changes. For these purposes, chickens were divided into four groups namely i) non supplemented and non-challenged (negative control, CON); ii) no supplementation of vit. E but challenged with ZJ1 (positive control, NS+CHA); iii) vit. E supplementation at the dose of 50 IU/day/Kg body weight and ZJ1 challenge (VE50+CHA); and 4) vit. E supplementation at the dose of 100 IU/day/Kg body weight and ZJ1 challenge (VE100+CHA). In all groups, we analyzed concentrations of glutathione (GSH), malondialdehyde (MDA), nitric oxide (NO), total antioxidant capacity (T-AOC), and activity of glutathione S-transferase (GST), superoxide dismutase (SOD), catalase (CAT) using biochemical methods. The virus loads were determined by quantitative RT-PCR and antibody titers by hemagglutination inhibition assays. We also examined the histopathological changes in the duodenal and jejunal mucosa at 3 and 5-day post infection (dpi) with NDV. Results: A significant elevation in the NO level was observed in NDV challenged chickens compared to the CON chickens at 2 dpi. The MDA contents were significantly increased whereas GSH was significantly decreased in NDV-challenged chickens compared to control. Furthermore, activities of GST, CAT, SOD, as well as the TOAC were markedly decreased in challenged chickens in comparison with control. Virus copy numbers were higher in NDV infected NS+CHA group compared to other groups. Severe histopathological changes including inflammation, degeneration and broken villi were observed in the intestine of NDV challenged chickens. However, all these malfunctions of antioxidant system and pathological changes in the intestine were partially or completely reversed by the vit. E supplementation. Conclusions: Our results suggest that NDV infection causes oxidative stress and histopathological changes in the duodenum and jejunum of chickens, which can be partially or fully ameliorated by supplementation of vit. E. Additionally, these findings suggest that oxidative stress contributes to the intestinal damages in NDV infected chickens. These findings will help to understand the pathogenesis of NDV and further investigation of therapeutic agents for control of Newcastle disease.

Ameliorative effects of selenium on the excess apoptosis of the jejunum caused by AFB1 through death receptor and endoplasmic reticulum pathways.

Aflatoxin B1 (AFB1), one of most potent and common mycotoxins in human food and animal feed, has hepatotoxic and carcinogenic effects on humans and poultry. Recent studies indicated that selenium (Se) has a protective effect on apoptosis induced by toxin poisoning. The present study was designed to reveal the ameliorative effects of selenium on the expression of apoptosis related molecules in the jejunum of broilers exposed to an AFB1 diet for 3 weeks. A total of 216 one-day-old healthy Cobb broilers were randomly divided into the control group (0 mg kg-1 AFB1), AFB1 group (0.6 mg kg-1 AFB1), AFB1 + Se group (0.6 mg kg-1 AFB1 + 0.4 mg kg-1 supplement Se) and Se group (0.4 mg kg-1 supplement Se), respectively. TUNEL and flow cytometry assays both indicated that 0.4 mg kg-1 selenium could ameliorate excess apoptosis caused by AFB1 in jejunal cells. Moreover, the expressions of FAS, FASL, TNF-α, TNF-R1, CASPASE-3, CASPASE-8, CASPASE-10, GRP78 and GRP94 analyzed by qRT-PCR demonstrated that 0.4 mg kg-1 selenium restored these parameters to be close to those in the control group. In summary, supplementation of selenium at a concentration of 0.4 mg kg-1 selenium could protect the chicken's jejunum from excess apoptosis caused by 0.6 mg kg-1 AFB1 through down-regulating the expression of death receptor pathway and endoplasmic reticulum pathway related molecules. According to this conclusion, this study may contribute to a better understanding of selenium's protective role against AFB1 poisoning.

Differential mRNA expression of nutrient transporters in male and female chickens.

Male chickens grow faster than female chickens, which may be due to greater nutrient uptake. The transport of nutrients from the intestine to the blood is mediated by transporters located on the surface of epithelial cells lining the villi. The objective of this study was to profile the mRNA expression of an aminopeptidase and selected amino acid and monosaccharide transporters in the duodenum, jejunum, and ileum of male and female chickens at d of hatch (doh) and at d 7 and d 14 post hatch. The mRNA abundance of aminopeptidase N (APN), a peptide (PepT1), 6 amino acid transporters (ASCT1, bo,+AT, CAT1, EAAT3, LAT1, and y+LAT2), and 3 monosaccharide (GLUT2, GLUT5, and SGLT1) transporters was assayed by real-time PCR. Data were analyzed by ANOVA using JMP Pro11. The abundance of bo,+AT, EAAT3, ASCT1, y+LAT2, and GLUT2 mRNA was greater in male than female chickens (P < 0.05). There was no difference in expression between males and females for the other 5 transporters and APN. There was a sex x age interaction for bo,+AT, PepT1, SGLT1, ASCT1, and y+LAT2 mRNA, with greater mRNA abundance in males than females at doh but no difference between males and females at d 7 and d 14. A 3-way sex x age x tissue interaction was observed for GLUT2 mRNA. There was greater GLUT2 mRNA abundance for males in the duodenum and ileum at doh and in the jejunum at d 7, but no difference between males and females at d 14. Thus, there was differential expression of some nutrient transporters in male and female chicks at doh but not at later ages.

Effect of feed supplementation with zinc glycine chelate and zinc sulfate on cytokine and immunoglobulin gene expression profiles in chicken intestinal tissue

The aim of the study was to evaluate the effect of inorganic and organic forms of Zn on the expression of cytokines (IL-2, TNF-α, IFN-γ, IL-12, IL-17, IL-4, IL-10, and TGF-β) and immunoglobulins (IgA and IgG) in the tissues of the small intestine (jejunum and ileum) of broiler chickens. In the experiment, 90 broiler chickens were divided into 4 experimental groups and a control group, with 18 birds each. The birds received Zn supplements in inorganic form with and without phytase (ZnSO4 and ZnSO4 + F), and in organic form with glycine, with and without phytase (Zn-Gly and Zn-Gly + F). The total rearing period was 42 days. Quantitative real-time (RT)-PCR was used to measure the expression of the cytokines and immunoglobulins. The differences between the results obtained for the control and experimental groups, between the groups receiving ZnSO4 and Zn-Gly, and between groups ZnSO4-F and Zn-Gly-F were analyzed statistically. High relative expression of IL-2 was observed for the chickens in the groups receiving ZnSO4-F, Zn-Gly, and Zn-Gly-F on d 42 in comparison to the control group. High relative expression of TNF-α, IL-12, and IL-17 was noted in the group that received ZnSO4 + F. High expression of IgG, IgA, IL-4, TGF-β, and IL-10 was noted in the groups of chickens that received feed supplemented with Zn-Gly and Zn-Gly + F chelates on d 42 of the study in comparison to the control group. In conclusion, supplementation with Zn-Gly chelates can ensure Th1 and Th2 balance during the immune response in the gut-associated lymphoid tissue (GALT), and, by increasing IgA and IgG expression, also can stimulate potentiation of the immune response involved in passive protection of the body from infection. In contrast, the use of inorganic forms of Zn, in the form of sulfates, can induce local inflammatory processes in the intestines, which, in the case of long-term supplementation, lead to the development of infections.

A study on the expression of apoptotic molecules related to death receptor and endoplasmic reticulum pathways in the jejunum of AFB1-intoxicated chickens.

Aflatoxin B1 (AFB1) is a common contaminant of poultry feeds in tropical and subtropical climates. Early researches have well established the hepatotoxic, carcinogenic, and immunotoxic effects of AFB1 on humans and animals. Recently, it has been shown that AFB1 could cause the up- or down-alteration of mitochondrial pathway molecule expression. However, the information on the expression of death receptor and endoplasmic reticulum molecules in the jejunal apoptosis induced by AFB1 were unavailable. So the present study was conducted to explore the expression of apoptotic molecules related to death receptor and endoplasmic reticulum in the jejunal cells of chickens exposed to AFB1 diet for 3 weeks. Total of 144 one-day-old chickens was randomly divided into two groups, namely control group (containing 0 mg/kg AFB1) and AFB1 group (containing 0.6 mg/kg AFB1). Histopathological observation and microscopic quantitative analysis revealed morphological changes in the jejunum such as the shedding of the mucosal epithelial cells in the apical region of villi along with the decrease of villus height, villus area and villus/crypt ratio in the AFB1 group. Both TUNEL and flow cytometry assays showed that AFB1 intake induced excessive apoptosis of jejunal cells. Quantitative real-time PCR test displayed the general upregulation of death receptors (FAS, FASL, TNF-α and TNF-R1), endoplasmic reticulum signals (GRP78 and GRP94) as well as initiator and executioner caspases (CASPASE-10, CASPASE-8 and CASPASE-3) in the jejunum of AFB1-intoxicated chickens. It's the first study demonstrating that AFB1 induced apoptosis of chickens’ jejunum accompanied by the alteration of death receptor and endoplasmic reticulum molecule expression.

Lysolecithin as feed additive enhances collagen expression and villus length in the jejunum of broiler chickens

Adding lysolecithin to feed has reportedly improved the performance of broiler chickens. Lysolecithin is generated by phospholipase catalyzed hydrolysis of lecithin. The enzymatic reaction converts various phospholipids into the corresponding lysophospholipids, with lysophosphatidylcholine (LPC) one of the primary products. Here we compared supplementation with a commercial lysolecithin (Lysoforte®) with comparable levels of highly purified LPC for effects on broilers. Despite no differences in weight gain during the starter period, we discovered a significant increase in average villus length with lysolecithin and an increase in villus width with purified LPC. High-throughput gene expression microarray analyses revealed many more genes were regulated in the epithelium of the jejunum by lysolecithin compared to purified LPC. The most up-regulated genes and pathways were for collagen, extracellular matrix, and integrins. Staining sections of the jejunum with Picrosirius Red confirmed the increased deposition of collagen fibrils in the villi of broilers fed lysolecithin, but not purified LPC. Thus, lysolecithin elicits gene expression in the intestinal epithelium, leading to enhanced collagen deposition and villus length. Purified LPC alone as a supplement does not mimic these responses. Feed supplementation with lysolecithin triggers changes in the intestinal epithelium with the potential to improve overall gut health and performance.

Expression of host defense peptides in the intestine of Eimeria-challenged chickens

Avian coccidiosis is caused by the intracellular protozoan Eimeria, which produces intestinal lesions leading to weight gain depression. Current control methods include vaccination and anticoccidial drugs. An alternative approach involves modulating the immune system. The objective of this study was to profile the expression of host defense peptides such as avian beta-defensins (AvBDs) and liver expressed antimicrobial peptide 2 (LEAP2), which are part of the innate immune system. The mRNA expression of AvBD family members 1, 6, 8, 10, 11, 12, and 13 and LEAP2 was examined in chickens challenged with either E. acervulina, E. maxima, or E. tenella. The duodenum, jejunum, ileum, and ceca were collected 7 d post challenge. In study 1, E. acervulina challenge resulted in down-regulation of AvBD1, AvBD6, AvBD10, AvBD11, AvBD12, and AvBD13 in the duodenum. E. maxima challenge caused down-regulation of AvBD6, AvBD10, and AvBD11 in the duodenum, down-regulation of AvBD10 in the jejunum, but up-regulation of AvBD8 and AvBD13 in the ceca. E. tenella challenge showed no change in AvBD expression in any tissue. In study 2, which involved challenge with only E. maxima, there was down-regulation of AvBD1 in the ileum, AvBD11 in the jejunum and ileum, and LEAP2 in all 3 segments of the small intestine. The expression of LEAP2 was further examined by in situ hybridization in the jejunum of chickens from study 2. LEAP2 mRNA was expressed similarly in the enterocytes lining the villi, but not in the crypts of control and Eimeria challenged chickens. The lengths of the villi in the Eimeria challenged chickens were less than those in the control chickens, which may in part account for the observed down-regulation of LEAP2 mRNA quantified by PCR. Overall, the AvBD response to Eimeria challenge was not consistent; whereas LEAP2 was consistently down-regulated, which suggests that LEAP2 plays an important role in modulating an Eimeria infection.

Efficacy of early chick nutrition with Aloe vera and Azadirachta indica on gut health and histomorphometry in chicks.

Aim:This study was conducted with an aim of studying the efficacy of water supplements of Aloe vera and Azadirachta indica (neem) during pre-starter age (0-2 weeks) on gut health and histomorphometry in Vanaraja chicks.Materials and Methods:A total of 192 day old Vanaraja chicks were randomly assigned to one of four herbal water treatments throughout the experimental pre-starter stage (0-2 weeks) in a completely randomized design. Each treatment was given four replicates consisting of 12 chicks per replicate. Water treatments comprised T1: Control with regular antibiotic supplement, T2: 3 ml Aloe juice per chick per day, T3: 3 ml neem extract per chick per day, T4: 1.5 ml Aloe and 1.5 ml neem per chick per day. Gut culture was done for Escherichia coli and Lactobacillus sps. and gut histomorphometry in 24 gut samples at 14 days of age.Results:This study revealed that supplementation of A. vera and neem in water significantly (p<0.05) reduced and increased the number of gut E. coli and Lactobacillus sps. Colonies, respectively, as compared to that of control groups; Villi was significantly (p<0.05) taller and broader on 14 days of age across the jejunum of chicks fed with neem supplementation as compared to that of control chicks. Significantly lower crypt depth (p<0.05) was observed in the duodenum of Aloe supplementation. Villus height: Crypt depth ratio of duodenum and jejunum was significantly (p<0.05) increased neem and Aloe supplementation in chicks as compared to their combination and control.Conclusion:Immediate post hatch supplementation of Aloe juice and neem extract in chicks improved the development and health of their gut.

Impact of thyme powder (Thymus vulgaris L.) supplementation on gene expression profiles of cytokines and economic efficiency of broiler diets.

This study was conducted in an attempt to evaluate the impact of thyme powder supplementation on broiler diets with respect to cytokine and mucin2 gene expressions. It was also our aim to evaluate the growth performance, blood biochemical and hematological profiles, and economic efficiency of the diet. A total of 120 1-dayold chicks (Cobb 500) were divided into four groups on the basis of the diet. One group received a basal diet (control) while the others received a basal diet supplemented with 2, 5, or 8g/kg of thyme powder. At 42days of age, the chickens were weighed and euthanized, and then blood and tissue samples were collected for the purpose of analysis. Results obtained clearly indicated that thyme supplementation of the diet, especially at 5g/kg, resulted in a significant increase in body weight and caused the increased body weight gain and feed intake (P˂0.05) as well as augmented WBC and lymphocyte count and IgG and NO levels (P˂0.001). The economic evaluation showed that birds fed on 8g/kg thyme yielded the lowest net revenue and highest feed cost to produce 1kg of live weight compared to the other treatments, while birds fed on 2 and 5g/kg thyme did not differ significantly from values obtained for the controls. The lipid profile of the broilers was affected by thyme supplementation at 2 and 5g/kg as represented by a significant decrease in serum cholesterol levels as well as low density lipoprotein levels, which is known to be associated with a corresponding increase in high-density lipoprotein (P˂0.001). Furthermore, supplementation of thyme downregulated the pro-inflammatory mediators and increased the expression of mucin2 mRNA in the jejunum of chickens. It can be concluded that thyme supplementation in the diet of broilers at 5g/kg has the potential to favorably influence productive performance via an improvement in the immune status of the broiler.

Impact of Fusarium mycotoxins on hepatic and intestinal mRNA expression of cytochrome P450 enzymes and drug transporters, and on the pharmacokinetics of oral enrofloxacin in broiler chickens

Cytochrome P450 (CYP450) drug biotransformation enzymes and multidrug resistance (MDR) proteins may influence drug disposition processes. The first part of the study aimed to evaluate the effect of mycotoxins deoxynivalenol (DON) and/or fumonisins (FBs), at contamination levels approaching European Union guidance levels, on intestinal and hepatic CYP450 enzymes and MDR proteins gene expression in broiler chickens. mRNA expression of genes encoding CYP450 enzymes (CYP3A37, CYP1A4 and CYP1A5) and drug transporters (MDR1/ABCB1 and MRP2/ABCC2) was determined using qRT-PCR. A significant up-regulation of CYP1A4 (P = 0.037) and MDR1 (P = 0.036) was observed in the jejunum of chickens fed a diet contaminated with FBs. The second part of this study aimed to investigate the impact of feeding a FBs contaminated diet on the oral absorption of enrofloxacin (10 mg/kg BW), a MDR1 substrate. A significant (P = 0.045), however small, decreased area under the plasma concentration-time curve (AUC0-48 h, mean ± SD) was observed for enrofloxacin in chickens fed the FBs contaminated diet compared to the control group, 16.28 ± 1.82 h μg/mL versus 18.27 ± 1.79 h μg/mL. These findings suggest that concurrent administration of drugs with FBs contaminated feed might alter the pharmacokinetic characteristics of CYP1A4 substrate drugs and MDR1 substrates, such as enrofloxacin.

Xanthophyll supplementation reduced inflammatory mediators and apoptosis in hens and chicks.

This study investigated effects of xanthophylls (containing 40% lutein and 60% zeaxanthin) on gene expression of inflammatory mediators ( [] and []) and apoptosis ( [] and ) of breeding hens and chicks. In Exp. 1, 432 hens were divided into 3 groups and fed diets supplemented with 0 (as the control group), 20, or 40 mg/kg xanthophylls. The liver, duodenum, jejunum, and ileum were sampled after 35 d. Results showed that 40 mg/kg of xanthophyll addition decreased in the liver, in the liver and duodenum, and in the liver and jejunum while increasing level in the liver and jejunum. Experiment 2 was a 2 × 2 factorial design. Male chicks hatched from hens fed 0 or 40 mg/kg xanthophyll diets were fed diets containing either 0 or 40 mg/kg xanthophylls. The liver, duodenum, jejunum, and ileum were sampled at 0, 7, 14, and 21 d after hatching. Results showed that in ovo xanthophylls reduced inflammatory mediators and apoptosis in the liver, duodenum, and jejunum of chicks mainly within 1 wk after hatching, whereas dietary xanthophylls only decreased expression in the liver from 2 wk onward. These results underlined important anti-inflammatory and antiapoptotic effects of maternal but not progeny dietary xanthophylls. In conclusion, xanthophylls can suppress inflammatory mediators and apoptosis in different tissues of hens and chicks.

10. The Effect of Chemically-Synthesized Silver Nanoparticles on Performance and the Histology and Microbiological Profile of the Jejunum in Chickens

Abstract The aim of the study was to analyse how per os application of hydrocolloids of silver nanoparticles (22 nm) and lipid-coated nanosilver hydrocolloids (5 nm) affect the microbiological status and morphology of the jejunum of broiler chickens and their growth performance. The experiment was conducted on 60 chickens. The first group was the control. The chickens in group II received a silver nanoparticle hydrocolloid (Ag-nano) at a dose of 5 mg/kg b.w./day. The chickens in group III received a lipid-coated nanosilver hydrocolloid (AgL-nano) at a dose of 5 mg/kg b.w./day. Samples of digesta were taken from the jejunum during dissection and the total numbers of fungi, aerobic bacteria and bacteria of the coli group were determined in the samples. Samples of the jejunum were also collected during dissection to determine the length of the villi and depth of the crypts. The silver nanoparticles had no effect on growth performance or the histological picture of the jejunum. An increase was noted in the total number of aerobic mesophilic bacteria and a decrease in the number of coli group bacteria, which are facultative anaerobes, which indicates that the nanosilver had a selective effect on the microflora of the digestive tract in the chickens.

Yeast β-d-glucans induced antimicrobial peptide expressions against Salmonella infection in broiler chickens

The present study was designed to investigate the effects of yeast β-d-glucans (YG) on gene expression of endogenous β-defensins (AvBDs), cathelicidins (Cath) and liver-expressed antimicrobial peptide-2 (LEAP-2) in broilers challenged with Salmonella enteritidis (SE). 240 day-old Cobb male broilers were randomly assigned to 2×2 factorial arrangements of treatments with two levels of dietary YG (0 or 200mg/kg in diet) and two levels of SE challenge (0 or 1×109 SE at 7–9 days of age). The results showed SE infection reduced growth performance,and increased salmonella cecal colonization and internal organs invasion, increased concentration of intestinal specific IgA and serum specific IgG antibody, as compared to uninfected birds. SE challenge differentially regulated AvBDs, Caths and LEAP-2 gene expression in the jejunum and spleen of broiler chickens during the infection period. However, YG supplementation inhibited the growth depression by SE challenge, and further increased level of serum specific IgG and intestinal specific IgA antibody. Higher level of salmonella colonization and internal organs invasion in the SE-infected birds were reduced by YG. SE-induced differentially expression patterns of AMPs genes was inhibited or changed by YG. Results indicated YG enhance chicken's resistance to salmonella infection.

Effect of ochratoxin A on the intestinal mucosa and mucosa-associated lymphoid tissues in broiler chickens.

The immunotoxic effect of ochratoxin A (OTA) on the intestinal mucosa-associated lymphoid tissue and its cytotoxic action on the intestinal epithelium were studied in broiler chickens experimentally treated with the toxin. From the 7th day of life, 80 male broiler chickens (Ross 308) were randomly divided into four groups of 20 birds each. The three experimental groups (E1-3) were treated with OTA for 28 days (E1: 50 μg/kg body weight [bw]/day; E2: 20 μg/kg bw/day; E3: 1 μg/kg bw/day) and the fourth group served as control. Histological examination of the intestinal mucosa and immunohistochemical staining for identification of CD4+, CD8+, TCR1 and TCR2 lymphocytes in the duodenum, jejunum and ileocaecal junction were performed, and CD4+/CD8+ and TCR1/TCR2 ratios were calculated. OTA toxicity resulted in decreased body weight gain, poorer feed conversion ratio, lower leukocyte and lymphocyte count, and altered intestinal mucosa architecture. After 14 days of exposure to OTA, immunohistochemistry showed a significant reduction of the lymphocyte population in the intestinal epithelium and the lamina propria. After 28 days of exposure, an increase in the CD4+ and CD8+ values in both the duodenum and jejunum of chickens in Groups E1 and E2 was observed, but the TCR1 and TCR2 lymphocyte counts showed a significant reduction. No significant changes were observed in Group E3. The results indicate that OTA induced a decrease in leukocyte and lymphocyte counts and was cytotoxic to the intestinal epithelium and the mucosa-associated lymphoid tissue, altering the intestinal barrier and increasing susceptibility to various associated diseases.

Expression of nutrient transporters in duodenum, jejunum, and ileum of Eimeria maxima-infected broiler chickens.

The uptake of amino acids is mediated by active transporters located on the basolateral and brush border membranes of intestinal epithelial cells. The current study investigated the expression of amino acid transporters (AAT) and other genes in the intestine of chicks infected with Eimeria maxima. At 7-day postinfection (PI), tissue from each intestinal segment (duodenum, jejunum, and ileum) was taken from birds inoculated with 3 × 10(3) oocysts/bird and processed to recover RNA. Analysis of gene expression was performed using real-time reverse transcription polymerase chain reaction (qRT-PCR). Results were given as relative expression using β₂-microglobulin as an endogenous control. All the genes studied were expressed in three segments of the intestines, and expression of the genes was altered by infection with E. maxima. Even though the jejunum is considered the parasite's primary predilection site, there was no segment-related difference in expression of most of the genes studied. The antimicrobial peptide (LEAP2) was downregulated in all three segments of the intestine. The results also demonstrate that transporters associated with brush border membranes were downregulated while transporters associated with the basolateral membranes were upregulated and that E. maxima alters the expression of AAT and LEAP2 throughout the small intestine.

Effects of heat stress on the gene expression of nutrient transporters in the jejunum of broiler chickens (Gallus gallus domesticus).

In broiler chickens, heat stress disrupts nutrient digestion and absorption. However, the underlying molecular mechanism is not clearly understood. Hence, to investigate the effects of high ambient temperatures on the expression levels of nutrient transporters in the jejunum of broiler chickens, seventy-two 35-day-old male broiler chickens with similar body weights were randomly allocated into two groups: control (24 ± 1 °C) and heat-stressed (32 ± 1 °C). The chickens in the heat-stressed group were exposed to 10 h of heat daily from 08:00 to 18:00 and then raised at 24 ± 1 °C. The rectal temperature and feed intake of the chickens were recorded daily. After 7 days, nine chickens per group were sacrificed by exsanguination, and the jejunum was collected. The results show that heat exposure significantly decreased the feed intake and increased the rectal temperature of the broiler chickens. The plasma concentrations of uric acid and triglyceride significantly increased and decreased, respectively, in the heat-stressed group. No significant differences in the levels of plasma glucose, total amino acids, and very low-density lipoprotein were observed between the heat-stressed and control groups. However, the plasma concentration of glucose tended to be higher (P = 0.09) in the heat-stressed group than in the control group. Heat exposure did not significantly affect the mRNA levels of Na(+)-dependent glucose transporter 1 and amino acid transporters y + LAT1, CAT1, r-BAT, and PePT-1. However, the expression levels of GLUT-2, FABP1, and CD36 were significantly decreased by heat exposure. The results of this study provide new insights into the mechanisms by which heat stress affects nutrient absorption in broiler chickens. Our findings suggest that periodic heat exposure might alter the jejunal glucose and lipid transport rather than amino acid transport. However, intestinal epithelial damage and cell loss should be considered when interpreting the effects of heat stress on the expression of intestinal transporters.

Cyclic AMP synergizes with butyrate in promoting β-defensin 9 expression in chickens

Host defense peptides (HDP) have both microbicidal and immunomodulatory properties. Specific induction of endogenous HDP synthesis has emerged as a novel approach to antimicrobial therapy. Cyclic adenosine monophosphate (cAMP) and butyrate have been implicated in HDP induction in humans. However, the role of cAMP signaling and the possible interactions between cAMP and butyrate in regulating HDP expression in other species remain unknown. Here we report that activation of cAMP signaling induces HDP gene expression in chickens as exemplified by β-defensin 9 (AvBD9). We further showed that, albeit being weak inducers, cAMP agonists synergize strongly with butyrate or butyrate analogs in AvBD9 induction in macrophages and primary jejunal explants. Additionally, oral supplementation of forskolin, an adenylyl cyclase agonist in the form of a Coleus forskohlii extract, was found to induce AvBD9 expression in the crop of chickens. Furthermore, feeding with both forskolin and butyrate showed an obvious synergy in triggering AvBD9 expression in the crop and jejunum of chickens. Surprisingly, inhibition of the MEK-ERK mitogen-activated protein kinase (MAPK) pathway augmented the butyrate-FSK synergy, whereas blocking JNK or p38 MAPK pathway significantly diminished AvBD9 induction in chicken macrophages and jejunal explants in response to butyrate and FSK individually or in combination. Collectively, these results suggest the potential for concomitant use of butyrate and cAMP signaling activators in enhancing HDP expression, innate immunity, and disease resistance in both animals and humans.

The effect of dietary supplementation of nitric oxide donor and inhibitor on nNOS expression in and motility of the small intestine of broilers

We investigated the effect of dietary supplementation of sodium nitroprusside (SNP), a nitric oxide (NO) donor, and N-nitro-L-arginine methyl ester (L-NAME), a NO inhibitor, on neuronal nitric oxide synthase (nNOS) expression in and motility of small intestinum in broilers. A total of 560, one-day-old Ross 308 hybrid mixed sex broiler chicks were divided randomly into one control and seven treatment groups for a 42 day feeding trial including starter phase (0–21 days) and grower phase (22–42 days). The control group was fed a basal diet and the experimental groups were the fed basal diet supplemented with 25, 50, 100 and 200 mg/kg SNP and 25, 50 or 100 mg/kg L-NAME. Ten chickens from each group were sacrificed to collect samples on days 21 and 42. The expression patterns of nNOS immunoreactivity in nerve fibers were determined by immunohistochemistry. In the contractility studies, longitudinal isolated strips of duodenum, jejunum and ileum were treated with 10−5 M L-arginine and 10−4 M SNP. Immunohistochemistry revealed that nNOS expression was not detectable in the duodenum or ileum of either the control or experimental groups. On the other hand, nNOS immunoreactivity in the jejunum control group showed a strong reaction on day 21, but the reaction was weak on day 42. nNOS expression clearly was suppressed on day 21 by the diet supplemented with L-NAME, while the diet supplemented with SNP stimulated nNOS expression on day 21. Contractility experiments revealed that spontaneous contractility of isolated strips of duodenum, jejunum and ileum showed no significant difference among groups. Spontaneous contractions of all strips were inhibited by L-arginine and SNP in all groups. The percentage inhibition rate of spontaneous contractions of jejunum application on days 21 and 42 after L-arginine decreased in the group supplemented with 100 mg/kg L-NAME. The percentage inhibition rate on day 21 after SNP application decreased in both groups that received 50 and 100 mg/kg L-NAME. We demonstrated the expression pattern of nNOS in nerve fibers in jejunum of broiler chickens. Contractility studies revealed that the NOS-NO pathway may play a role in smooth muscle contraction of small intestine of chickens. Feeding strategies that supplement NO donor and NO inhibitor can be of physiological importance to small intestine motility owing to alteration of nNOS expression in the jejunum.

Effect of Citric Acid and Microbial Phytase on Small Intestinal Morphology in Broiler Chicken

An experiment was carried out to investigate the effects of citric acid (CA) (0, 3 and 6%) and microbial phytase (MP) (0, 500 and 1000 U/kg) on morphology of different segments of small intestine (duodenum, jejunum and ileum) in broiler chickens fed on corn and soybean meal based diets. The effect of 9 experimental treatments (3×3 factorial design) were assessed using 270 7-d-old Ross 308 male broiler chicks in a randomized complete block design in three replicates of 10 birds each. The mean villi length (VL), crypt depth (CD) and goblet cell number (GCN) in duodenum, jejunum and ileum and villi width (VW) and VL:CD ratio in duodenum was significantly greater for the birds fed on acidified diets compared to the control birds at day 42 of age (P<0.01). Inclusion of 3% CA in diet significantly decreased the epithelial thickness (ET) in duodenum, jejunum and ileum (P<0.01). The birds received diets with 1000 U/kg of MP showed significant increase in CD (P<0.01) and GCN in jejunum (P<0.05), and significant decrease in VL:CD ratio and ET in the duodenum (P<0.01), jejunum (P<0.05) and ileum (P<0.01) segments. No variable of interest were affected by CA × MP interaction. It was concluded that CA and MP independently exhibit positive impact on morphometery of small intestine, toward facilitating the nutrient absorption and reducing the metabolic demands of the intestinal tract in broiler chickens.