Effects of Immune Stress on Performance Parameters, Intestinal Enzyme Activity and mRNA Expression of Intestinal Transporters in Broiler Chickens

Article information

Asian-Australas J Anim Sci. 2012;25(5):701-707
Publication date (electronic) : 2012 May 01
doi : https://doi.org/10.5713/ajas.2011.11377
1College of Animal Science and Technology, Northwest A & F University, Shaanxi, Yangling 712100, China
2Department of Life Science, Shanxi Agricultural University, Taigu 030801, China.
*Corresponding Author: Junhu Yao. Tel: +86-29-87092102, Fax: +86-29-87092164, E-mail: yaojunhu2004@sohu.com
aThese authors contribute equally to this work.
Received 2011 October 17; Revised 2012 February 13; Accepted 2012 January 11.


Immune stress is the loss of immune homeostasis caused by external forces. The purpose of this experiment was to investigate the effects of immune stress on the growth performance, small intestinal enzymes and peristalsis rate, and mRNA expression of nutrient transporters in broiler chickens. Four hundred and thirty-two 1-d-old broilers (Cobb500) were randomly assigned to four groups for treatment; each group included nine cages with 12 birds per cage. Group 1 = no vaccine (NV); Group 2 = conventional vaccine (CV); group 3 = lipopolysaccharide (LPS)+conventional vaccine (LPS); group 4 = cyclophosphamide (CYP)+conventional vaccine (CYP). The results demonstrated that immune stress by LPS and CYP reduced body weight gain (BWG), feed intake (FI), small intestine peristalsis rate and sIgA content in small intestinal digesta (p<0.05). However, the feed conversion ratio (FCR) remained unchanged during the feeding period. LPS and CYP increased intestinal enzyme activity, relative expression of SGLT-1, CaBP-D28k and L-FABP mRNAs (p<0.05). LPS and CYP injection had a negative effect on the growth performance of healthy broiler chickens. The present study demonstrated that NV and CV could improve growth performance while enzyme activity in small intestine and relative expression of nutrient transporter mRNA of NV and CV were decreased in the conditions of a controlled rational feeding environment. It is generally recommended that broilers only need to be vaccinated for the diseases to which they might be exposed.


It is apparent that many factors can influence the immune response of an animal under stress. Stress can suppress, enhance or have no effect on the immune response of an animal. There are many interacting factors influencing the immunological response of an animal under stress, these include stressor types (psychological vs physiological vs physical), duration of stressor (chronic vs acute), genetics, age, and immune status. Some factors in the homeostasis of broilers directly or indirectly affect their immune status, and immunosuppression induced by stress is manifested by failures in vaccination and increased morbidity and mortality of flocks (Shini et al., 2010). Vaccinations are considered necessary to reduce the mortality rate that occurs in modern poultry production. However, immunizations have sometimes been administered in large doses at a high frequency and improper vaccination may lead to immune stress. Studies have demonstrated that stress could affect the intestinal function in animals (Apajalathi and Kettunen, 2003; Teo and Tan, 2007; Rajkumar et al., 2011; Wilkinson et al., 2011) and furthermore disturb the absorption of nutrients (Garriga et al., 2006; Jacobi et al., 2006; Albin et al., 2007). The activities of digestive enzymes are such important factors affecting the absorption and utilization of nutrients in the alimentary tract that they are often used to indicate the intestine function of nutrient absorption. Heat stress causes metabolic and physiological changes in the intestinal transport of glucose through the increase of SGLT1 (Carriga et al., 2006). The effects of stress on the intestinal nutrient absorption in animals may be reflected in two ways. One is nonspecific, in which the absorption of nutrients is affected by the changes of digestive enzymes and intestinal peristalsis rate (Tsukada et al., 2002); the other is specific, in which the absorption of nutrients is affected by changing the expression of transporters particularly of nutrients (Shepherd et al., 2004). Expression of these proteins varies during the development of chickens at different ages (Gilbert et al., 2007; Duarte et al., 2011; Yadgary et al., 2011). Research has shown that the immune response can affect growth performance and enteric diseases in commercial poultry, contribute to losses in productivity, increase mortality, and increase the contamination of products for human consumption (Yang et al., 2011). Previous studies have yielded various results; however, the mechanisms of these phenomena have not yet been fully understood. To our knowledge, few studies on the potential effect on the status of broilers reared under immune stress conditions were available. The aims of this study were to determine the balance between the hypo- and hyper-stress, minimize distress while increasing conditions for eustress.


Animals and experimental design

The experiment was performed as a single-factor trial. Four hundred thirty-two 1-d-old broilers (Cobb500) were randomly assigned to four treatments with nine pens per treatment and twelve birds per pen. The four treatments were control group with no vaccine (NV), conventional vaccine group (CV), lipopolysaccharide injected+ conventional immunization group (LPS), and cyclophosphamide injected+conventional immunization group (CYP). All birds were housed in an environmentally controlled room with an initial room temperature of 34°C for the first 5 d and gradually decreased to 21°C by the end of the experiment at the third week. Birds were provided with feed and water ad libitum. The lighting program was 24 h light for the first day, followed by 18 h light/6 h dark for the rest of the trial period. All experimental procedures were approved by the Animal Ethics Committee of the Northwest A & F University.

Chemicals and vaccines

The chemicals used in this study included LPS from E. coli (L2880, Sigma-Aldrich, Inc., St. Louis, Missouri, USA) and CYP (C0768, Sigma-Aldrich). The vaccines used in this study included infectious bronchitis vaccine (IBH 120), the inactivated avian influenza (AI) vaccine (H5N2, N28 strain), the live vaccine strain Clone-30 of the Newcastle disease virus (NDV), the combined inactive vaccines for infectious bursal disease and the Newcastle disease (ND-IB), all of them were supplied by Harbin Veterinary Research Institute (Harbin, China).

Growth performance measurement

Body weight gain (BWG) and feed intake (FI) were recorded based on cage weekly. Mortality was recorded daily and feed conversion ratio (FCR) was corrected according to mortality.

Sample collection

Six birds with body weight close to the mean from each replicate were killed by cervical dislocation on d 21 and d 42. The duodenum, jejunum and ileum were weighed. Digesta from the duodenum, jejunum and ileum was put in ice-cold phosphate buffered saline (PBS, pH 7.4). Samples were centrifuged (13,000×g for 5 min) at 4°C, and supernatant was frozen in a 2 ml eppendorf tube immediately at −20°C until needed. The activities of amylase, lipase and trypsin in the small intestinal digesta were determined using standard kits (Jiancheng Bioengineering Institute, Nanjing, China). Intestinal peristalsis rate was detected according to the method described by Poggioli et al. (1995). The level of digesta sIgA was examined with the chicken IgA ELISA quantitation kit (no. E30–103, Behyl Laboratories Inc., Montgomery, TX, USA) as mentioned by Schuijffel et al. (2005).

RNA extraction and real-time PCR (RT-PCR)

Total RNA was extracted from individual intestinal tissue (duodenum, jejunum and ileum) using the RNeasy kit (Kim et al., 2010) (Qiagen, Germantown, MD, USA). The samples from small intestine were removed and deposited in a −80°C freezer for further analyses of SGLT1, CaBP-D28k and L-FABP. A 20 to 30 mg aliquot of each sample was weighed and kept on dry ice until homogenization according to manufacturer’s recommendation. In the extraction, RNA was eluted by rinsing the column membrane twice with 50 μl of RNase-free water. Total RNA concentration was determined by optical density (OD) 260 (NanoDrop-1000, Thermo Fisher Scientific, Waltham, MA, USA), and RNA purity was verified by OD260/OD280 value and formaldehyde-agarose gel electrophoresis. The first strand cDNA was stored at −20°C. The relative abundance of transporter mRNA was assayed by quantitative RT-PCR (Table 1).

Primer sequences for PCR amplification of target genes

Gene expression of SGLT-1, CaBP-D28k and L-FABP was assessed by 2-step quantitative RT-PCR using the SYBR Green PCR master mix (Applied Bio Systems Inc., Foster City, CA); mRNA expression was quantified using the iCycler iQTM5 multicolor RT-PCR detection system (Bio-Rad Laboratories, Hercules, CA, USA). A 25 μl PCR mixture was set up as follows: cDNA, 1 μl; each primer (10 pmol/ml), 1 μl; 2×SYBR Green quantitative PCR master mix, 12.5 μl; and double-distilled water, 10.5 μl. The temperature cycles were set as following: 95°C for 30 s, followed by 40 cycles at 95°C for 5 s and 60°C for 30 s. SYBR green fluorescence was measured at the end of each cycle. The melting curve was recorded at 60°C. All primer concentrations were optimized before actual runs. All measurements were carried out in triplicate to obtain the average values. Expression of SGLT-1, CaBP-D28k, and L-FABP mRNAs from different parts of small intestine was calculated by the 2−ΔΔCt method (Livak and Schmittgen, 2001).

Statistical analysis

Means of replicate were used as the experimental unit for statistical analysis. Data were subjected to one-way ANOVA analysis for the differences between the four groups using SAS software (SAS Institute Inc., Cary, North Carolina, USA, 2002). Differences between means at p< 0.05 were considered to be statistically significant.


Performance parameters

The effect of different types of immune stress on feed intake (FI), body weight gains (BWG) and the feed conversion ratio (FCR) is presented in Table 2. No differences were observed in FI and BWG between NV group and CV group (p>0.05). However, compared with NV, the BWG decreased significantly in the chickens that had been treated with LPS or CYP (p<0.05 and p<0.01), and the FI decreased significantly in the chickens that had been treated with LPS. The FCR showed no significant difference among the four groups (p>0.05).

Effect of different immune stressors on growth performance in broilers1

Measurements of organ relative weight, enteric enzyme and intestinal peristaltic rate

Relative weight indices of duodenum and jejunum in NV group were higher than that in LPS group on d 21 (p<0.05). There was no significant difference in the duodenum, jejunum and ileum weight indices on d 42 (Table 3).

Effects of immunity stress on small intestinal index in broilers (mg/g)

As shown in Table 4, the amylase and lipase of jejunum and ileum in CYP treatment were higher compared with the NV and CV treatments on d 21 and 42 (p<0.05 and p<0.01), while the trypsin activity in the small intestine was not significantly different during the whole period (p>0.05). The intestinal peristalsis rate and sIgA content in NV group were significantly higher than those in LPS and CYP groups (p<0.01) (Table 5).

Effects of immunity stress on digestive enzyme activity in small intestinal digesta (U/mg)

Effects of immunity stress on intestinal motility and digesta sIgA in broilers

Effects of stress on the expression of SGLT1, CaBP-D28k and L-FABP mRNAs in small intestine

The mRNA levels of related genes in the small intestine are shown in Table 6. The relative expression of SGLT-1, CaBP-D28k and L-FABP mRNAs in the three parts of small intestine in the LPS and CYP groups was significantly or numerically up-regulated in comparison to CV and NV groups (p<0.05).

Effects of immunity stress on the relative expression of SGLT1, CaBP-D28k and L-FABP mRNAs in the different parts of intestine


Our results showed that immune stress decreased the FI and BWG of chickens (Table 2). It was reported that immune stress may decrease the growth performance of broilers (Eid et al., 2003; Lin et al., 2004; Virden et al., 2007; Star et al., 2008; Rajapakse et al., 2010). The depressed growth performance might be explained by which immune stress is the immuno-modulator and allocates nutrients toward immune response.

Dudley et al. (1998) observed that dietary restriction leads to a decrease in intestinal protein synthesis with concomitant lowering of intestinal weight. Thermal stress caused a consistent increase in amylase activity in broilers, which could result in a significant increase in starch hydrolysis (Osman et al., 1983). Other studies, however, reported that thermal stressed birds had a higher enzymatic activity (Routman et al., 2003). Furthermore, compared to birds fed with control diets, birds with early feed restriction and exogenous protease and amylase supplementation have increased maltase, lipase, and trypsin immediately after the restriction period (Pinheiro et al., 2004). In fact, birds subjected to feed restriction will increase intestinal absorption, and the gastrointestinal tract will be spared the effect of restriction on weight that affects other organs and the BWG (Pinheiro et al., 2004). These responses depend on the age of animal and the duration of the restriction period (Casirola et al., 1997; Ferraris et al., 2001). These results may indicate the adaptation of the enzymatic response to immune stress, the higher enzyme activity may have been stimulated by the presence of feed in the gastrointestinal tract for a longer time due to a slower rate of feed passage through the gastrointestinal tract (Barash et al., 1992). Moreover, some investigators e.g. Lessire et al. (1990) observed that feed restriction tended to improve protein, fat, and starch digestibilities irrespective of the diet. However, slower movement speed can increase the chyme residence time in the gut and help nutrient absorption. The current research demonstrated that the effect of immune stress on chickens increased the stress stimulation in the intestinal tract, which might lead to the increased production of intestinal amylase and lipase enzymes in the LPS and CYP groups.

The intestine is relatively enriched with cells actively secreting IgA, and sIgAs from the lamina propria of villi in the duodenum and jejunum, and Peyer’s patches are important components of intestinal mucosal immunity (Mestecky et al., 1987). Mucosal immunity is an important part of the humoral immunity. Secretory IgA is the most prominent antibody present at mucosal surfaces, and is the factor of mucosal immunity that provides passive immunoprotection against invading pathogens in the gastrointestinal tract. The results of this study showed that injection with LPS and CYP could decrease sIgA content compared with the NV group. This implies that LPS and CYP may impair the humoral immune system, which led to small intestinal mucosal lesions.

Nutrient transporters play a critical role in the absorption of luminal substrate. In the small intestine of animals, SGLT1 is the key factor that affects absorption of glucose in the small intestine (Kellett, 2001; Wood and Frayhum, 2003). A previous study showed that glucocorticoids released during stress periods induce the expression of glucocorticoid regulated kinase, which enhances glucose transportation by increasing SGLT1 abundance in the cell membrane (Carriga et al., 2006). Gilbert et al. (2007) also observed an increase in mRNA abundance of SGLT1 in feed restricted chickens. SGLT1 has the unique ability to transport glucose against its concentration gradient, and it is considered the major pathway for glucose assimilation in the small intestine. Thus, a lower luminal sugar concentration than that in blood during feed restriction may cause the increase in this transporter, due to its important role in the assimilation of glucose. Calcium is absorbed in the duodenum and jejunum by calcium-binding protein (CaBP), which is regulated by a 28 KDa 1, 25(OH)-D3 protein (CaBP-D28k) (Craviso et al., 1987). Two types of fatty acid binding protein (FABP) are found in intestinal epithelium, i. e. intestinal FABP (I-FABP) and liver FABP (L-FABP) (Banaszak et al., 1994). I-FABP participates in the uptake of long-chain fatty acids from digesta in the small intestine into intestinal epithelial cells, and L-FABP transports fatty acids from the cells to the tissues (Prows et al., 1995). In the present study, the increasing demand for energy causes changes in the intestinal epithelium and gene expression of nutrient transporters. The upregulated expression of CaBP-D28k and L-FABP genes in stressed birds may be related to an increased intestinal absorptive function due to immune stress. The increased expression of nutrient transporter mRNA during stress is likely to be a compensation for the loss of absorptive area in the small intestine.

It can be concluded that broilers with NV or CV can achieve higher growth performance in a controlled, rational feeding environment, while immune stress can interrupt the homeostasis of the intestine and impair intestinal mucosal immune function. However increased enzyme and mRNA expression in chickens under stress might compensate for the nutrient consumption by the system immune responses or basic metabolic requirements. Extra vaccinating will cost extra growth. This study provides some details about the relationship between immune stress and nutrient partition.


The authors are grateful to the support from the Science & Technological Projects of Shaanxi Province, China (2010ZDGC-02, 2011KTCQ02-02), and National Natural Science Foundation of China (No.31001017 and No.31172223).


Albin DM, Wubben JE, Rowlett JM, Tappenden KA, Nowak RA. 2007;Changes in small intestinal nutrient transport and barrier function after lipopolysaccharide exposure in two pig breeds. J Anim Sci 85:2517–2523.
Apajalathi J, Kettunen A. 2003. Analysis and dietary modulation of the microbial community in the avian gastrointestinal tract. In : Proceedings of the 26th Technical Turkey Conference, Turkeys Magazine. p. 49–55.
Banaszak L, Winter N, Xu Z. 1994;Lipid-binding protein: a family of fatty acid and retinoid transport protein. Adv Protein Chem 45:89–151.
Craviso GL, Garrett KP, Clemens TL. 1987;1, 25-Dihydroxy vitamin D3 induces the synthesis of vitamin D-dependent calcium-binding protein in cultured chick kidney cells. Endocrinology 120:894–902.
Dudley MA, Wykes LJ, Dudley AW Jr, Burrin DG, Nichols BL, Rosenberger J, Jahoor F, Heird WC, Reeds PJ. 1998;Parenteral nutrition selectively decreases protein synthesis in the small intestine. Am J Physiol 274:G131–137.
Duarte CRA, Vicentini-Paulino MLM, Buratini J Jr, Castilho ACS, Pinheiro DF. 2011;Messenger ribonucleic acid abundance of intestinal enzymes and transporters in feed-restricted and refed chickens at different ages. Poult Sci 90:863–868.
Eid YZ, Ohtsuka A, Hayashi K. 2003;Tea polychenols reduce glucocorticoid induced growth inhibition and oxidative stress in broiler chickens. Br Poult Sci 44:127–132.
Gal-Garber O, Mabjeesh SJ, Sklan D, Uni Z. 2000;Partial sequence and expression of the gene for and activity of the sodium glucose transporter in the small intestine of fed, starved and refed chickens. J Nutr 130:2174–2179.
Garriga C, Hunter RR, Amat C, Planas JM, Mitchell MA, Moretó M. 2006;Heat stress increases apical glucose transport in the chicken jejunum. Am J Physiol Regul Integr Comp Physiol 290:R195–201.
Gilbert ER, Li H, Emmerson DA, Webb KE Jr, Wong EA. 2007;Developmental regulation of nutrient transporter and enzyme mRNA abundance in the small intestine of broilers. Poult Sci 86:1739–1753.
Jacobi SK, Gabler NK, Ajuwon KM, Davis JE, Spurlock ME. 2006;Adipocytes, myofibers, and cytokine biology: new horizons in the regulation of growth and body composition. J. Anim. Sci. 84(E-Suppl):E140–E149.
Kellett GL. 2001;The facilitated component of intestinal glucose absorption. J Physiol 531:585–595.
Kim S, Miska KB, Jenkins MC, Fetterer RH, Cox CM, Stuard LH, Dalloul RA. 2010;Molecular cloning and functional characterization of the avian macrophage migration inhibitory factor (MIF). Dev Comp Immunol 34:1021–1032.
Lemmens JWTJ. 1995;Nucleic acid levels, cellular activity and growth during the puerulus stage of the Western Pock Lobster (Panulirus Cygnus(George); Decapoda: Palinuridae). J Exp Mar Biol Ecol 194:143–156.
Lessire M. 1990;Effect of feeding technique, ad libitum, dry or wet force feeding on the metabolisable energy values of raw materials for poultry. Br Poult Sci 31:785–793.
Lin H, Decuypere E, Buyse J. 2004;Oxidative stress induced by corticosterone administration in broiler chickens (Gallus gallus domesticus) 1. Chronic exposure. Comp. Biochem. Physiol 139B:737–744.
Livak KJ, Schmittgen TD. 2001;Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408.
Mestecky J, Czerkinsky C, Russell MW, Brown TA, Prince SJ, Moldoveanu Z, Jackson S, Michalek SM, McGhee JR. 1987;Induction and molecular properties of secretory and serum IgA antibodies specific for environmental antigens. Ann Allergy 59:54–59.
Mitchell MA, Carlisle AJ. 1992;The effect of chronic exposure to elevated environmental temperature on intestinal morphology and nutrient absorption in the domestic fowl (Gullus domesticus). Comp. Biochem. Physiol. 101A:137–142.
Nasir A, Moudgal RP, Singh NB. 1999;Involvement of corticosterone in food intake, food passage time and in vivo uptake of nutrients in the chicken (Gallus domesticus). Br Poult Sci 40:517–522.
Osman AM, Tanios NI. 1983;The effect of heat on the intestinal and pancreatic levels of amylase and maltase of laying hens and broilers. Comp. Biochem. Physiol. 75A:563–567.
Pinheiro DF, Cruz VC, Sartori JR, Vicentini Paulino ML. 2004;Effect of early feed restriction and enzyme supplementation on digestive enzyme activities in broilers. Poult Sci 83:1544–1550.
Poggioli R, Benelli A, Arletti R, Cavazzuti E, Bertolini A. 1995;K+ channel openers delay intestinal transit and have antidiarrheal activity. Eur J Pharmacol 287:207–209.
Prows DR, Murphy EJ, Schroeder F. 1995;Intestinal and liver fatty acid-binding proteins differentially affect fatty acid uptake and esterification in L-cell. Lipids 30:907–910.
Rajapakse JRPV, Buddhika MDP, Nagataki M, Nomura H, Watanabe Y, Ikeue Y, Agatsuma T. 2010;Effect of sophy beta-glucan on immunity and growth performance in broiler chicken. J Vet Med Sci 72:1629–1632.
Rajkumar U, Reddy MR, Rao SVR, Radhika K, Shanmugam M. 2011;Evaluation of growth, carcass, immune response and stress parameters in naked neck chicken and their normal siblings under tropical winter and summer temperatures. Asian-Aust J Anim Sci 24:509–516.
Routman KS, Yoshida L, Frizzas de Lima AC, Macari M Jr, Pizauro JM. 2003;Intestinal and pancreas enzyme activity of broilers exposed to thermal stress. Brazilian J Poult Sci 5:23–27.
Savory J. 1986;Influence of ambient temperature on feeding activity parameters and digestive function in domestic fowls. Physiol Behav 38:353–357.
Institute SAS. 2002. SAS users guide: Statistics SAS Institute Inc. Cary, NC, USA:
Schuijffel DF, van Empel PC, Pennings AM, vanPutten JP, Nuijten PJ. 2005;Passive immunization of immune-suppressed animals: Chicken antibodies protect against Ornithobacterium rhinotracheale infection. Vaccine 23:3404–3411.
Shini S, Huff GR, Shini A, Kaiser P. 2010;Understanding stress induced immunosuppression: Exploration of cytokine and chemokine gene profiles in chicken peripheral leukocytes. Poult Sci 89:841–851.
Shepherd EJ, Helliwell PA, Mace OJ, Morgan EL, Patel N, Kellett GL. 2004;Stress and glucocorticoid inhibit apical Glut2-trafficking and intestinal glucose absorption in rat small intestine. J Physiol 560:281–290.
Star L, Kemp B, van den Anker I, Parmentier HK. 2008;Effect of single or combined climatic and hygienic stress in four layer lines: 1. Performance Poult Sci 87:1022–1030.
Teo AY, Tan HM. 2007;Evaluation of the performance and intestinal gut micro flora of broilers fed on corn-soy diets supplemented with Bacillus subtilis PB6 (CloSTAT). J Appl Poult Res 16:296–303.
Tsukada F, Sawamura K, Kohno H, Ohkubo Y. 2002;Mechanism of inhibition of small intestinal motility by restraint stress differs from that with norepinephrine treatment in rats. Biol Pharm Bull 25:122–124.
Uni Z, Tako E, Gal-Garber O, Sklan D. 2003;Morphological, molecular, and functional changes in the chicken small intestine of the late-term embryo. Poult Sci 82:1747–1754.
Virden WS, Lilburn MS, Thaxton JP, Corzo A, Hoehler D, Kidd MT. 2007;The effect of corticosterone induced stress on amino acid digestibility in Ross broilers. Poult Sci 86:338–342.
Wilkinson KGE, Tee RB, Tomkins G, Premier R. 2011;Effect of heating and aging of poultry littern the persistence of enteric bacteria. Poult Sci 90:10–18.
Wilson EK, Pierson EW, Hester PY, Adam RL, Stadelman WJ. 1980;The effect of high environment temperature on food passage tissue and performance traits of Pekin ducks. Poult Sci 59:2322–2330.
Wood S, Frayhum P. 2003;Glucose transporters (GLUT and SGLT): expanded families of sugar transport protein. Br J Nutr 89:3–9.
Yang XJ, Li WL, Feng Y, Yao JH. 2011;Effects of immune stress on growth performance, immunity, and cecal microflora in chickens. Poult Sci 49:2740–2746.
Yadgary L, Yair R, Uni Z. 2011;The chick embryo yolk sac membrane expresses nutrient transporter and digestive enzyme genes. Poult Sci 90:410–416.

Article information Continued

Table 1

Primer sequences for PCR amplification of target genes

Target genes Primer sequences Predicted size (bp) Genebank accession
113 XM_415247
163 NM 204192.1
116 NM 205513.1
86 NM_205518

Table 2

Effect of different immune stressors on growth performance in broilers1

Item2 Weeks Treatments SEM1 p value

BWG (kg/bird) 1–3 0.70 0.69 0.64 0.68 0.010 0.166
4–6 1.53d 1.44c 1.37a 1.41b 0.015 0.001
1–6 2.23c 2.13b 2.01a 2.09b 0.022 0.001
FI (kg/bird) 1–3 0.90 0.90 0.91 0.89 0.659 0.355
4–6 2.90b 2.79ab 2.69a 2.70ab 0.152 0.026
1–6 3.80b 3.69ab 3.59a 3.60a 0.329 0.004
FCR 1–3 1.29 1.31 1.41 1.32 0.031 0.418
4–6 1.89 1.93 1.97 1.93 0.032 0.103
1–6 1.59 1.62 1.69 1.63 0.009 0.184

Each value represents the mean of 9 replicates for each treatment.


BWG = Body weight gain; FI = Feed intake; FCR = Feed conversion ratio.


Means within the same column with different superscripts are significantly different (p<0.05). NV = No vaccine; CV = Conventional vaccine; LPS = Lipopolysaccharide; CYP = Cyclophosphamide.

Table 3

Effects of immunity stress on small intestinal index in broilers (mg/g)

Day Item Treatments SEM1 p value

21 d Duodenum 0.95b 0.90b 0.84a 0.86ab 0.015 0.006
Jejunum 1.34b 1.21b 1.05a 1.17ab 0.026 0.005
Ileum 1.05 1.10 1.26 1.19 0.039 0.232
42 d Duodenum 0.54 0.51 0.56 0.48 0.014 0.383
Jejunum 1.44 1.31 1.29 1.12 0.046 0.281
Ileum 1.15 1.00 1.10 1.00 0.045 0.720

Each value represents the mean of 9 replicates for each treatment.


Means within the same row column with different superscripts are significantly different (p<0.05).

NV = No vaccine; CV = Conventional vaccine; LPS = Lipopolysaccharide; CYP = Cyclophosphamide.

Table 4

Effects of immunity stress on digestive enzyme activity in small intestinal digesta (U/mg)

Enzyme Item Treatments SEM1 p value

21 d Amylase Duodenum 187.91 140.30 220.59 233.52 15.237 0.147
Jejunum 180.64a 186.61a 371.01b 364.16b 23.460 0.001
Ileum 160.85a 198.49ab 228.80bc 282.33c 12.240 0.003
Lipase Duodenum 353.00a 381.51a 654.03b 563.74ab 38.192 0.027
Jejunum 459.67a 616.91bc 628.85bc 744.96c 27.481 0.008
Ileum 416.65a 510.35ab 625.65bc 708.08c 28.987 0.004
Trypsin Duodenum 79.15 75.70 80.94 86.61 1.558 0.284
Jejunum 85.23 90.92 101.34 99.64 1.913 0.460
Ileum 82.30 87.18 93.24 95.42 3.647 0.444
42 d Amylase Duodenum 130.77 127.15 137.17 149.59 8.026 0.811
Jejunum 131.18a 151.62ab 194.21bc 213.94c 9.148 0.006
Ileum 119.61a 174.98ab 188.33b 193.73b 11.552 0.045
Lipase Duodenum 188.53 196.47 264.74 298.66 16.000 0.084
Jejunum 339.01a 361.93a 670.34b 627.05b 35.595 0.001
Ileum 233.08a 305.66ab 362.42bc 415.50c 21.731 0.025
Trypsin Duodenum 147.31 152.54 157.68 168.43 5.563 0.656
Jejunum 134.03 138.59 161.92 173.90 6.006 0.241
Ileum 118.38 143.89 152.47 160.74 7.590 0.213

Each value represents the mean of 9 replicates for each treatment.


Means within the same column with different superscripts are significantly different (p<0.05).

NV = No vaccine; CV = Conventional vaccine; LPS = Lipopolysaccharide; CYP = Cyclophosphamide.

Table 5

Effects of immunity stress on intestinal motility and digesta sIgA in broilers

Item Treatments SEM1 p value

21 d peristalsis rate (%) 0.662c 0.624c 0.532a 0.586b 0.017 0.001
21 d sIgA content (mg/g) 472.00c 458.00c 420.76b 356.37a 3.570 0.001
42 d sIgA content (mg/g) 464.15b 457.70b 428.24a 387.23a 6.277 0.001

Each value represents the mean of 9 replicates for each treatment.


Means within the same row with different superscripts are significantly different (p<0.05).

NV = No vaccine; CV = Conventional vaccine; LPS = Lipopolysaccharide; CYP = Cyclophosphamide.

Table 6

Effects of immunity stress on the relative expression of SGLT1, CaBP-D28k and L-FABP mRNAs in the different parts of intestine

Day Gene name Item Treatments SEM1 p value

21 d SGLT1 Duodenum 0.89a 1.04a 2.02b 2.64b 0.664 0.006
Jejunum 0.98a 0.99a 1.33ab 1.58b 0.397 0.008
Ileum 0.81 0.78 1.02 1.01 0.477 0.380
CaBP-D28k Duodenum 1.12 0.84 1.17 1.89 0.124 0.068
Jejunum 1.02ab 0.93a 1.40b 1.45b 0.174 0.002
Ileum 0.87 0.97 1.38 1.54 0.953 0.099
L-FABP Duodenum 1.00a 0.90a 1.65c 1.41b 0.777 0.001
Jejunum 0.86 0.66 1.22 1.32 0.951 0.131
Ileum 1.03a 1.21ab 2.26c 1.95bc 0.182 0.007
42 d SGLT1 Duodenum 1.42b 0.73a 1.60c 1.97c 0.137 0.004
Jejunum 0.78a 0.92ab 1.50c 1.29bc 0.893 0.040
Ileum 0.84 0.95 1.22 1.05 0.660 0.400
CaBP-D28k Duodenum 1.24ab 0.99a 2.31c 1.85bc 0.157 0.006
Jejunum 1.33a 1.14a 2.46b 2.40b 0.178 0.001
Ileum 1.06 1.06 1.85 1.96 0.138 0.065
L-FABP Duodenum 1.14ab 0.85a 1.96b 2.02b 0.176 0.050
Jejunum 1.28ab 0.82a 1.86bc 2.10c 0.155 0.020
Ileum 1.27 0.81 1.31 1.53 0.106 0.087

Each value represents the mean of 9 replicates for each treatment.


Means within the same row with different superscripts are significantly different (p<0.05).

NV = No vaccine; CV = Conventional vaccine; LPS = Lipopolysaccharide; CYP = Cyclophosphamide.