Impacts of increasing challenge with Eimeria maxima on the growth performance and gene expression of biomarkers associated with intestinal integrity and nutrient transporters.

This study was conducted to investigate the impacts of graded severity of Eimeria maxima infection on the growth performance and intestine health of broiler chickens. Four different levels of E. maxima-challenged treatments were used, including a non-challenged control group, a low challenge (12 500 oocysts), a medium challenge (25 000 oocysts), and a high challenge dose (50 000 oocysts). There were eight replicate cages per treatment, with 12 birds in each cage, and chickens in the challenged groups orally received sporulated oocysts on day 14. Gastrointestinal permeability was measured by fluorescein isothiocyanate dextran at 5 days post-infection (dpi), whereas intestinal morphology and gene expression of nutrient transporters and tight junction proteins were determined at 6 dpi. The results demonstrate a linear reduction in growth performance, jejunal villus height, and jejunal integrity with graded challenge doses of E. maxima (P < 0.01). Moreover, linear regulation of nutrient transporters and tight junction proteins was a consequence of increasing Eimeria infection levels (P < 0.01). The linear increase of Claudin 1, cationic amino acid transporter, glucose transporter 1, and L-type amino acid transporter genes was associated with increased severity of coccidiosis (P < 0.01). Furthermore, expression of nutrient transporters located at the brush border membrane were down-regulated (P < 0.01) with increasing E. maxima inoculation dose. In conclusion, growth performance and key intestinal integrity biomarkers in broiler chickens were adversely influenced in a dose-dependent manner by E. maxima infection.

Effects of multi-strain probiotic supplementation on intestinal microbiota, tight junctions, and inflammation in young broiler chickens challenged with Salmonella enterica subsp. enterica.

OBJECTIVE: This study assessed the effects of probiotics on cecal microbiota, gene expression of intestinal tight junction proteins, and immune response in the cecal tonsil of broiler chickens challenged with Salmonella enterica subsp. enterica. METHODS: One-day-old broiler chickens (n = 240) were randomly allocated to four treatments: negative control (Cont), multi-strain probiotic-treated group (Pro), Salmonella-infected group (Sal), and multi-strain probiotic-treated and Salmonella-infected group (ProSal). All chickens except those in the Cont and Pro groups were gavaged with 1×108 cfu/mL of S. enterica subsp. enterica 4 days after hatching. RESULTS: Our results indicated that body weight, weight gain, and feed conversion ratio of birds were significantly reduced (p<0.05) by Salmonella challenge. Chickens challenged with Salmonella decreased cecal microbial diversity. Chickens in the Sal group exhibited abundant Proteobacteria than those in the Cont, Pro, and ProSal groups. Salmonella infection downregulated gene expression of Occludin, zonula occludens-1 (ZO1), and Mucin 2 in the jejunum and Occludin and Claudin in the ileum. Moreover, the Sal group increased gene expression of interferon-γ (IFN-γ), interleukin-6 (IL-6), IL-1ß, and lipopolysaccharide-induced tumor necrosis factor-alpha factor (LITAF) and reduced levels of transforming growth factor-ß4 and IL-10 compared with the other groups (p<0.05). However, chickens receiving probiotic diets increased Lactobacillaceae abundance and reduced Enterobacteriaceae abundance in the ceca. Moreover, supplementation with probiotics increased the mRNA expression of Occludin, ZO1, and Mucin 2 in the ileum (p<0.05). In addition, probiotic supplementation downregulated the mRNA levels of IFN-γ (p<0.05) and LITAF (p = 0.075) and upregulated IL-10 (p = 0.084) expression in the cecal tonsil. CONCLUSION: The administration of multi-strain probiotics modulated intestinal microbiota, gene expression of tight junction proteins, and immunomodulatory activity in broiler chickens.

Different methionine to cysteine supplementation ratios altered bone quality of broilers with or without Eimeria challenge assessed by dual energy X-ray absorptiometry and microtomography.

Despite the acknowledged significance of nutrition in bone development, effects of methionine (Met) and cysteine (Cys) on bone quality remain under-researched, particularly during Eimeria challenge. We investigated the effects of different supplemental Met to Cys ratios (MCR) on bone quality of broilers under Eimeria challenge. A total of 720 fourteen-day old Cobb500 broilers were allocated into a 5 × 2 factorial arrangement. Five diets with Met and Cys supplemented at MCR of 100:0, 75:25, 50:50, 25:75, and 0:100 were fed to the birds with or without Eimeria challenge. Body composition was measured by dual energy x-ray absorptiometry, and the femur bone characteristics were assessed by microtomography. Data were analyzed by two-way ANOVA and orthogonal polynomial contrast. The results reaffirmed the detrimental effects of Eimeria challenge on bone quality. On 9 d post inoculation (DPI), significant interaction effects were found for whole body bone mineral content (BMC), lean tissue weight, and body weight (P < 0.05); in the nonchallenged group (NCG), these parameters linearly decreased as MCR decreased (P < 0.05). In the challenged group (CG), body weight and lean tissue weight were unaffected by MCR, and BMC linearly increased as MCR decreased (P < 0.05). For the cortical bone of femoral metaphysis on 6 DPI, bone mineral density (BMD) linearly increased as MCR decreased (P < 0.05). Bone volume to tissue volume ratio (BV/TV) in the CG linearly increased as MCR decreased (P < 0.05). On 9 DPI, BMC and TV linearly increased as MCR decreased (P < 0.05) in the NCG. BMD and BV/TV changed quadratically as MCR decreased (P < 0.05). For the trabecular bone of femoral metaphysis on 9 DPI, BV/TV, and trabecular number linearly increased as MCR decreased (P < 0.05) in the NCG. For the femoral diaphysis, BV, TV, BMC on 6 DPI, and BMD on 9 DPI linearly increased as MCR decreased (P < 0.05). In conclusion, this study showed that both Eimeria challenge and varying supplemental MCR could influence bone quality of broilers.

Impacts of varying methionine to cysteine supplementation ratios on growth performance, oxidative status, intestinal health, and gene expression of immune response and methionine metabolism in broilers under Eimeria spp. challenge.

A study was conducted to investigate effects of different methionine (Met) to cysteine (Cys) supplementation ratios (MCR) on growth performance, oxidative status, intestinal health, immune responses, and methionine metabolism in broilers under Eimeria challenge. A total of 720 male Cobb500 broilers (14-day-old) were allocated in a 2 × 5 factorial arrangement (5 diets, with or without challenge) with 6 replicates per treatment. The total sulfur amino acid concentrations were consistent across treatments meeting the breeder's recommendation, only MCR varied. The diets were labeled as MET100; MET75; MET50; MET25; and MET0, representing MCR of 100:0; 75:25; 50:50; 25:75; and 0:100, respectively. Data were analyzed by 2-way ANOVA and orthogonal polynomial contrast. Growth performance declined linearly or quadratically as MCR decreased (P < 0.01). On 6-day postinoculation (DPI), interaction effects (P < 0.01) were found; BW and body weight gain were lower in MET0 compared to the other treatments in the nonchallenged groups, whereas not in the challenged groups. On 6 and 9 DPI, serum total antioxidant capacity linearly decreased as MCR decreased (P < 0.05). Hepatic activities of glutathione peroxidase on 6 DPI and superoxide dismutase on 9 DPI changed quadratically as MCR decreased (P < 0.05). The digestibility of Met linearly decreased whereas the digestibility of Cys linearly increased as MCR decreased. The ileal crypt depth linearly decreased as MCR decreased (P < 0.01) on 6 DPI. The expression of transforming growth factor beta on 6 and 9 DPI, tumor necrotic factor alpha and interleukin 10 on 9 DPI changed quadratically as MCR decreased (P < 0.05). Eimeria challenge increased expression of Met adenosyltransferase and cystathionine gamma-lyase, whereas decreasing the expression of other Met metabolism genes (P < 0.01) on 6 DPI. Expression of Met metabolism genes linearly increased as MCR decreased (P < 0.05). In conclusion, different Met to Cys supplementation ratios exerted linearly or quadratically effects on the growth performance, oxidative status, intestinal health, and metabolism of Met in broiler chickens under Eimeria infection.

Reduced bone formation and increased bone resorption drive bone loss in Eimeria infected broilers.

Coccidiosis is an economically significant disease in the global poultry industry, but little is known about the mechanisms of bone defects caused by coccidiosis; thus, the study focused on effects of coccidiosis on the bone homeostasis of young broiler chickens. A total of 480 male Cobb500 broilers were randomly allocated into four treatment groups, including an uninfected control consuming diet ad libitum, two infected groups were orally gavaged with two different concentrations of sporulated Eimeria oocysts, and an uninfected pair-fed group fed the same amount of feed as the high Eimeria-infected group consumed. Growth performance and feed intake were recorded, and samples were collected on 6 days post infection. Results indicated that coccidiosis increased systemic oxidative status and elevated immune response in bone marrow, suppressing bone growth rate (P < 0.05) and increasing bone resorption (P < 0.05) which led to lower bone mineral density (P < 0.05) and mineral content (P < 0.05) under Eimeria infection. With the same amount of feed intake, the uninfected pair-fed group showed a distinguished bone formation rate and bone resorption level compared with the Eimeria infected groups. In conclusion, inflammatory immune response and oxidative stress in broilers after Eimeria infection were closely associated with altered bone homeostasis, highlighting the role of inflammation and oxidative stress in broiler bone homeostasis during coccidiosis.

Effects of levels of methionine supplementations in forms of L- or DL-methionine on the performance, intestinal development, immune response, and antioxidant system in broilers challenged with Eimeria spp.

The study was conducted to investigate the effects of 2 isoforms of methionine on growth performance and intestinal health induced by methionine (Met) deficiency and Eimeria infection in broilers. A total of 720 one-day old male chicks (Cobb500) were randomly allocated to 10 groups in a 2 × 5 factorial arrangement (6 reps/group, 12 birds/cage) with diets and Eimeria challenge as the main factors. Hundred percent DL-Met, 100% L-Met, 80% DL-Met, and 80% L-Met diets were formulated to meet approximately 100 or 80% of the total sulfur amino acid (TSAA) requirement with DL-Met or L-Met as Met supplementation sources. The 60% TSAA basal diet (60% Met) was formulated without Met supplementation. At d14, the challenge groups were gavaged with mixed Eimeria spp. Growth performance was recorded on d7, 14, 20 (6-day postinfection [DPI]), and 26 (12 DPI). The gut permeability was measured on 5 and 11 DPI. Antioxidant status and gene expression of immune cytokines and tight junction proteins were measured on 6 and 12 DPI. Data were analyzed by 1-way and 2-way ANOVA before and after the challenge, respectively. Orthogonal polynomial contrasts were used for post hoc comparison. Overall, the Eimeria challenge and 60% Met diet significantly reduced growth performance, antioxidant status, and mRNA expression of tight junction genes and immune cytokines. For other Met treatments, the L-Met groups had significantly higher BWG and lower FCR than the DL-Met group from d 1 to 20. The L-Met groups had less gut permeability than the DL-Met groups on 5 DPI. Compared to the 80% Met groups, the 100% Met groups reduced gut permeability. At 6 DPI, the 80% Met groups showed higher ZO1 expression than the 100% Met groups. The challenge groups had higher Muc2 expression and GSH/GSSG compared to the nonchallenge groups, and SOD activity was lower in the L-Met groups compared to the DL-Met groups at 6 DPI. The 100% Met groups had higher GPx activity than the 80% Met groups at 12 DPI. In conclusion, during coccidiosis, the 100% Met groups had better gut integrity and antioxidant status. Met supplementation in the form of L-Met improved growth performance in the starter phase and gut permeability in the challenge phase.

The effects of xylo-oligosaccharides on regulating growth performance, nutrient utilization, gene expression of tight junctions, nutrient transporters, and cecal short chain fatty acids profile in Eimeria-challenged broiler chickens.

A 21-d experiment was conducted to investigate the effects of xylo-oligosaccharides (XOS) on growth performance, nutrient utilization, gene expression of tight junctions, nutrient transporters, and cecal short chain fatty acids (SCFA) profile of broiler chickens challenged with mixed Eimeria spp. Two hundred fifty-two zero-day-old chicks were allocated to 6 treatments in a 3 × 2 factorial arrangement (corn-soybean meal diets supplemented with 0, 0.5, or 1.0 g/kg XOS; with or without Eimeria challenge). Challenged groups were inoculated with a solution containing E. maxima, E. acervulina, and E. tenella oocysts on d 15. During the infection period (d 15 to d 21), there was a significant (P < 0.05) Eimeria × XOS interaction for weight gain (WG). XOS significantly (P < 0.05) increased WG in the unchallenged birds but not in the challenged treatments. There was no significant Eimeria × XOS interaction for N and minerals utilization responses. XOS supplementation at 0.5 g/kg tended to alleviate Eimeria-induced depression in apparent ileal digestibility of DM (P = 0.052). Challenged birds had lower (P < 0.01) AME, AMEn, and total retention of N, Ca, and P. Eimeria upregulated (P < 0.01) gene expression of tight junction proteins claudin-1, junctional adhesion molecule-2, and glucose transporter GLUT1; but downregulated (P < 0.01) the peptide transporter PepT1, amino acid transporters rBAT, CAT2, y+LAT2, and zinc transporter ZnT1. XOS alleviated (P < 0.05) Eimeria-induced claudin-1 upregulation. Eimeria decreased (P < 0.05) cecal saccharolytic SCFA acetate, butyrate, and total SCFA, but increased (P < 0.05) branched chain fatty acids isobutyrate and isovalerate. The supplementation of XOS tended to decrease the concentration of isobutyrate (P = 0.08) and isovalerate (P = 0.062). In conclusion, 0.5 g/kg XOS supplementation alleviated depression in growth performance and nutrient utilization from the Eimeria challenge. In addition, supplemental XOS reversed the gene expression changes of claudin-1, also showed the potentials of alleviating the negative cecal fermentation pattern induced by Eimeria infection.

Effects of Tannic Acid Supplementation on Growth Performance, Oocyst Shedding, and Gut Health of in Broilers Infected with Eimeria Maxima.

The purpose of this study was to evaluate effects of tannic acid (TA) on growth performance, fecal moisture content, oocyst shedding, gut permeability, lesion score, intestinal morphology, apparent ileal digestibility, and the antioxidant and immune system of broilers infected with Eimeria maxima. A total of 420 one-day-old broilers were distributed to five treatments with seven replicates of 12 birds. The five treatments were the (1) sham-challenged control (SCC; birds fed a control diet and administrated with PBS); (2) challenged control (CC; birds fed a control diet and inoculated with E. maxima); (3) tannic acid 0.5 (TA0.5; CC + 500 mg/kg TA); (4) tannic acid 2.75 (TA2.75; CC + 2750 mg/kg TA); and (5) tannic acid 5 (TA5; CC + 5000 mg/kg TA). The TA2.75 group had significantly lower gut permeability compared to the CC group at 5 days post-infection (dpi). Supplementation of TA linearly reduced oocyst shedding of E. maxima at 7 to 9 dpi (p < 0.05). At 13 dpi, the TA2.75 group had significantly greater apparent ileal digestibility (AID) of dry matter (DM) and organic matter (OM) compared to the CC group. At 13 dpi, supplementation of TA linearly increased jejunal villus height (VH). Thus, this study showed that supplementation of TA at levels of 500 to 2750 mg/kg has the potential to be an anti-coccidial agent against E. maxima in broilers.

Nutrient profile and effects of carinata meal as alternative feed ingredient on broiler performance, tight junction gene expression and intestinal morphology.

Two studies were conducted to establish carinata meal as a partial replacement of conventional protein sources. Study I was conducted to determine the nutrient profile, nitrogen-corrected true metabolizable energy (TMEn), and amino acid (AA) digestibility of 2 groups: low glucosinolate carinata meal (LGCM) and high glucosinolate carinata meal (HGCM) using rooster assays. The LGCM contained 28 µmol/g glucosinolate, 11.5% moisture, 39.2% crude protein (CP), whereas the HGCM had 100 µmol/g glucosinolate, 10.1% moisture, 39.5% CP on as is basis. The precision-fed rooster assays were conducted to determine TMEn and AA digestibility. The TMEn levels of LGCM and HGCM were 1,814 and 1,690 kcal/kg on as is basis, respectively. Standardized digestibility for lysine, methionine, cysteine, threonine, and valine were 72, 88, 69, 75, and 79% for LGCM and 80, 89, 71, 76, and 80% for HGCM, respectively. Based on the nutrient profiles from study I, study II was conducted to evaluate the effects of LGCM and HGCM in broilers. A total of 504 one-day-old Cobb500 male broiler chickens were randomly divided into 42 battery cages with 6 replicates of 12 birds per cage. The seven dietary treatments were control diet, 3 inclusion levels of LGCM (4, 8, and 12%), and 3 of HGCM (4, 8, and12%) in a corn-SBM based diet fed for 21 d. No significant differences in BW, BWG, and FI were observed except for significantly lower BWG in 12% HGCM group compared to control for 14-21 days (P < 0.05). The FCR for 12% HGCM increased significantly compared to 4 and 8% of both LGCM and HGCM groups during wk 3 (14-21 d). Based on these studies, carinata meal could be recommended to partially replace conventional feed ingredients at a rate of 12% when LGCM is used and 8% when HGCM is used with no deleterious effects on growth performance, gut histology, and tight junction proteins.

Effects of phytase and coccidial vaccine on growth performance, nutrient digestibility, bone mineralization, and intestinal gene expression of broilers.

A study was conducted to evaluate effects of phytase and coccidial vaccine on growth performance, bone mineralization, nutrient digestibility, and intestinal gene expression of broiler chickens. The experiment was conducted in a 2 × 4 completely randomized factorial arrangement with 6 replicates per treatment and 10 birds each. Applications of coccidiosis vaccine and different dietary treatments were the 2 main factors in the current study. The dietary treatments included 1) a positive control (PC; 0.90% Ca and 0.45% available P: avP); 2) a negative control (NC; 0.75% Ca and 0.30% AvP); 3) NC + 500 FTU/kg of phytase (NC + 500PHY); and 4) NC + 1500 FTU/kg of phytase (NC + 1500PHY). Data were analyzed using SAS by 2-way ANOVA via GLM procedure. The statistical significance was set at P ≤ 0.05, and means were further separated using Tukey's Test. The results indicated that vaccination had no effect on growth performance except for feed intake from 0 to 14 d but negatively (P < 0.05) regulated bone ash and Ca digestibility. Birds fed with the Ca and P-reduced diet (NC) showed a lower BWG and bone ash compared to birds fed with the normal diet (PC), but supplementing phytase mitigated the negative effects on those birds. Broilers fed the NC diet had higher (P < 0.05) total Ca and P digestibility, and phytate degradation; supplementing phytase further increased P digestibility and phytate degradation of the broilers. A significant interaction (P < 0.05) between phytase and vaccination was observed, suggesting the vaccinated birds fed the PC diet and the unvaccinated birds fed the NC + 1500PHY increased calcium-sensing receptor gene expression compared with the unvaccinated birds fed the PC diet. In conclusion, in spite of coccidiosis vaccine, supplementing phytase at 1,500 FTU/kg alleviated the negative effects on growth performance, bone mineralization, and apparent ileal digestibility of P and phytate.

Roles of Nitrocompounds in Inhibition of Foodborne Bacteria, Parasites, and Methane Production in Economic Animals.

Nitrocompounds are derivatives of hydrocarbons, alcohols, fatty acids, and esters, consisting one or more nitro functional groups. Either natural sources of nitrocompounds or synthetic chemicals have been applied in animal diets to investigate their effects on economic animals, since conjugates of 3-nitropropanol and 3-nitropropionic acid were isolated from Astragalus oblongifolius. In this review, emphasis will be placed on nitrocompounds' antimicrobial activity, toxicity, metabolisms and mechanisms of actions. Nitrocompounds can be metabolized by ruminal microbials, such as Denitrobacterium detoxificans, or alcohol dehydrogenase in the liver. Moreover, it has been found that nitrocompounds are capable of inhibiting pathogens, parasites, methane and ammonia production; however, overdose of nitrocompounds could cause methemoglobinemia or interfere with energy production in mitochondria by inhibiting succinate dehydrogenase.

Effects of combination of mannan-oligosaccharides and ß-glucan on growth performance, intestinal morphology, and immune gene expression in broiler chickens.

The study was conducted to evaluate the effects of the combination of mannan-oligosaccharides (MOS) and ß-glucan on growth performance, intestinal morphology, and immune gene expression in broiler chickens. A total of 640, one-day-old male Cobb 500 broilers were randomly allocated into 32 pens with 8 replicates and 20 birds per pen. Thirty-two pens were divided into 4 treatments, including control, 0.04% MOS, 0.002% ß-glucan, and a combination of MOS and ß-glucan. Growth performance was measured on d14, 28, and 35. The ileum and cecal tonsils were collected from one bird per pen at 21 and 35 d of age for further analyses of immune gene expression. Duodenum, jejunum, and ileum were collected for intestinal morphology on d 35. Results indicated that both MOS and ß-glucan improved growth performance during starter phase (P < 0.05). In addition, ß-glucan further increased body weight gain of birds from d 0 to 28 (P < 0.05). Furthermore, the combination of MOS and ß-glucan presented higher villi height in the jejunum on d 35 (P < 0.05). There were no significant differences for gene expressions of immune responses on d 21 and 35. In conclusion, the application of prebiotic combination of MOS and ß-glucan might perform multiple pathways, improving growth performance in broiler chickens.

Assay considerations for fluorescein isothiocyanate-dextran (FITC-d): an indicator of intestinal permeability in broiler chickens.

Fluorescein isothiocyanate-dextran (FITC-d) is being used as an indicator of intestinal paracellular permeability in poultry research. Especially with the industry moving toward antibiotic-free production, intestinal function and integrity issues have been a research focus. An increasing number of scientific conference abstracts and peer-reviewed journal publications have shown that 4-kDa FITC-d is an efficient marker candidate for measurement of intestinal permeability and can be applied in broiler research. However, experimental protocols vary by personnel, instruments used, and research institution, and potential concerns related to this assay have yet to receive the same amount of attention. Understanding protocol consistency within and across laboratories is vital for obtaining accurate, consistent, and comparable experimental results. This review is aimed to 1) summarize different FITC-d assays in broiler research from peer-reviewed publications during the past 6 yr and 2) discuss factors that can potentially affect intestinal permeability results when conducting the FITC-d assay. In summary, it is essential to pay attention to details, including gavage dose, fasting period, sample handling and lab analysis details when conducting the assay in broiler research. Differences in birds (breed/strain, age, and gender) and experimental design (diet, health status/challenge model, and sampling age) need to be considered when comparing serum FITC-d concentration results between different in vivo animal trials.

Evaluating endogenous loss and standard ileal digestibility of amino acids in response to the graded severity levels of E. maxima infection.

The study was conducted to evaluate the effects of graded levels of Eimeria maxima challenge on endogenous loss, apparent ileal digestibility (AID), and standard ileal digestibility (SID) of amino acids. A total of 768 fourteen-day-old male Cobb 500 broiler chickens were randomly allocated into 64 battery cages. Apart from the regular corn-soybean based diet, the nitrogen-free diet (NFD) was formulated to determine the endogenous loss of amino acids. One-half of the birds (32 cages) were fed the NFD, and another half fed the regular diet from d 14 to 20. Both groups were further assigned to 4 treatments (nonchallenged control or three levels of challenge doses) with 8 replicate cages. The challenge doses were: the low challenge dose (Low) with E. maxima 12,500 oocysts, the medium challenge dose (Medium) with 25,000 E. maxima oocysts, and the high challenge dose (High) with 50,000 E. maxima oocysts. At 6 d postinfection, ileal digesta samples were collected and the intestinal lesion score were recorded. The results indicated a significant linear increase of endogenous amino acid flow in response to the graded E. maxima challenge. Moreover, the AID and SID of amino acids were linearly reduced due to the increasing challenge dose. The study demonstrated that NFD significantly reduced lesion scores, underestimating the true endogenous losses of birds fed regular diets. Even though the endogenous loss of amino acids was underestimated, they were linearly increased in response to the graded E. maxima challenge. In conclusion, the higher Eimeria dose birds were challenged with, the more endogenous amino acids were released into the intestine and the lower dietary nutrients were digested and absorbed by broiler chickens.

Effects of low-crude protein diets supplemented with arginine, glutamine, threonine, and methionine on regulating nutrient absorption, intestinal health, and growth performance of Eimeria-infected chickens.

The study was conducted to evaluate the effects of low crude protein diets supplemented with arginine, glutamine, methionine, and/or threonine on apparent ileal digestibility of amino acids, intestinal morphology, intestinal permeability, gene expression of nutrient transporters, and tight junction proteins of broiler chickens challenged with mixed Eimeria spp. A total of five hundred seventy-six, 12-day-old male broiler chickens were allocated into 8 treatments, and 6 replicate cages of 12 chickens per cage. This experiment included a nonchallenged control (NC) fed regular corn-soybean meal-based diet (Regular diet, 19% crude protein), an Eimeria-challenged control (CC) fed Regular diet, an Eimeria challenge group fed low-crude protein diet (LCP, 16% crude protein), 4 Eimeria challenge groups fed low-crude protein diet supplemented with 0.75% arginine, glutamine, methionine, and threonine, respectively (ARG, GLN, MET, and THR), and an Eimeria challenge group fed low-crude protein diet with 0.75% supplemented arginine, glutamine, methionine, and threonine collectively as a combination group (COMB). On d 14, birds in the challenge groups were gavaged with a mixed Eimeria spp. solution containing 12,500 oocysts of E. maxima, 12,500 oocysts of E. tenella, and 62,500 oocysts of E. acervulina. The results showed that the Eimeria challenge reduced overall growth performance, but the LCP had no adverse impacts on intestinal health and growth of Eimeria-infected birds compared to the CC. Additionally, supplementation of crystalline arginine, glutamine, methionine, and threonine improved the apparent ileal digestibility of these specific amino acids on 6 dpi. Moreover, the THR treatment increased villus height in the duodenum. The ARG treatment decreased intestinal permeability and gene expression of amino acid transporters, whereas the GLN and THR treatments both reversed adverse effects of coccidiosis on gene expression of tight junction protein (claudin 1). However, the MET and COMB treatments exacerbated infection severity of coccidiosis. In summary, adding 0.75% of arginine, glutamine, or threonine in a low crude protein diet can improve the intestinal health of birds challenged with a mild coccidia infection.

Effects of Eimeria tenella Infection on Key Parameters for Feed Efficiency in Broiler Chickens.

The purpose of the study was to investigate effects of different inoculation dosages of E. tenella on growth performance, gastrointestinal permeability, oocyst shedding, intestinal morphology, fecal consistency, ileal apparent digestibility, antioxidant capacity, and cecal VFA profile in broiler chickens. Five different dosages (T0: 0, T1: 6250, T2: 12,500, T3: 25,000, and T4: 50,000) of E. tenella oocysts were inoculated via oral gavage to fourteen-day-old broilers. Inoculation of E. tenella linearly increased FCR (p < 0.05), and feed intake was quadratically increased on 6 days post-infection (dpi; p = 0.08) and 7 dpi (p = 0.09). Cecal lesion score of each treatment was T0: 0; T1: 0.39 ± 0.14; T2: 0.93 ± 0.21; T3: 1.25 ± 0.16; and T4: 1.58 ± 0.2. Cecal total VFA production was linearly reduced due to E. tenella infection on 6 dpi (p < 0.01). E. tenella infection deepened cecal crypts depth on 6 dpi (CD; p < 0.05). Gastrointestinal permeability tended to be linearly increased (p = 0.07). E. tenella infection tended to linearly reduce duodenal VH (p = 0.1) and jejunal VH on 9 dpi (p = 0.09). Different dosages of E. tenella modulated the tendency of fecal moisture content and oocyst shedding. Therefore, E. tenella infection impaired feed efficiency and small intestinal health mainly by reducing cecal VFA production and deepening cecal CD in broilers.

Evaluation of nitro compounds as feed additives in diets of Eimeria-challenged broilers in vitro and in vivo.

Coccidiosis is a disease caused by Eimeria spp., resulting in approximately 3 billion US dollar loss in the poultry industry annually. The present study evaluated the effects of potential feed additives, 2-Nitro-1-propanol (NP) and nitroethanol (NE), on control of coccidiosis. An in vitro experiment indicated that both NP and NE inhibited the development of sporozoites in Madin-Darby bovine kidney cells (MDBK). The in vivo study was further conducted to evaluate the effects of NP and NE on growth performance, nitrogen-corrected apparent metabolizable energy (AMEn), and intestinal lesion scores of broilers challenged with Eimeria spps. Six treatments were tested in the study, including the nonchallenged control, challenged control, 100 ppm NP, 200 ppm NP, 100 ppm NE, and 200 ppm NE. Broilers were fed the treatment diets from day 12 until the end of the trial. All birds except the unchallenged control were challenged with Eimeria maxima, Eimeria tenella, and Eimeria acervulina on day 14. The growth performance was calculated, and the intestinal lesion was scored on day 20. The results showed that Eimeria challenge significantly reduced growth performance, increased intestinal lesion scores, and decreased AMEn compared with the nonchallenged control group. Birds fed with 200 ppm of NP had reduced growth performance compared with the nonchallenged control and challenged control. However, the supplementation of NP significantly improved AMEn and reduced cecal damage. Overall, NP and NE reduced sporozoites numbers in the MDBK cells. NP improved dietary digestibility of energy and reduces lesion scores in the ceca but could not maintain growth performance in broiler chickens infected with Eimeria spp.

Graded Eimeria challenge linearly regulated growth performance, dynamic change of gastrointestinal permeability, apparent ileal digestibility, intestinal morphology, and tight junctions of broiler chickens.

This study was conducted to evaluate graded Eimeria challenge on growth performance, apparent ileal digestibility, gastrointestinal permeability, intestinal morphology, gene expression of tight junction protein, and intestinal lesion scores in broiler chickens. There were 5 groups in this study, including a control and 4 different Eimeria treatment doses. A mixed Eimeria spp. solution with 50,000 Eimeria maxima, 50,000 Eimeria tenella, and 250,000 Eimeria acervulina per milliliter was prepared for the high-dose challenge treatment. The 2-fold serial dilution was used to make the medium-high (25,000 E. maxima; 25,000 E. tenella; 125,000 E. acervulina), the medium-low (12,500 E. maxima; 12,500 E. tenella; 62,500 E. acervulina), and the low challenge dose (6,250 E. maxima; 6,250 E. tenella; 31,250 E. acervulina). A total of three hundred sixty 13-day-old male broiler chickens were randomly allocated into 5 treatments with 6 replicated cages. Growth performance was calculated from 0 to 6 D postinfection (DPI). Intestine lesion was scored on 6 DPI. Gastrointestinal permeability was measured on 3, 5, 6, 7, and 9 DPI. The results indicated significant linear reduction in growth performance, intestinal villi height, and ileal nutrient digestibility in response to the increase of Eimeria challenge dose. Furthermore, gene expression of tight junction protein was linearly upregulated by the increasing challenge doses. Significant linear increases of gastrointestinal permeability were found on 5, 6, and 7 DPI (P < 0.01). On 9 DPI, the gastrointestinal permeability was recovered back to normal level in the challenge groups. In conclusion, the higher Eimeria doses birds received, the more severe intestine damage was observed in several gastrointestinal health parameters. The medium-low or medium-high levels of mixed Eimeria oocysts is suggested as an optimum Eimeria-challenge dose to establish a subclinical challenge model for future studies evaluating nutritional strategies. Moreover, it is recommended to measure gastrointestinal permeability on 5 DPI with higher oocysts doses and 6 DPI when using the lower oocysts doses.

2-Nitro-1-propanol improved nutrient digestibility and oocyst shedding but not growth performance of Eimeria-challenged broilers.

A 2 × 3 factorial arrangement study was conducted to evaluate 3 dosages of 2-nitro-1-propanol (NP; 0, 150, and 200 ppm) on intestinal health of birds with or without Eimeria challenge. A total of 432 thirteen-day-old male broiler chickens were randomly allocated to 6 treatments with 8 replicate cages of 9 birds per cage. All birds were fed with treatment diets from day 13 to 21. Birds in the challenge groups were gavaged with Eimeria maxima (50,000 oocysts per bird), Eimeria tenella (50,000 oocysts per bird), and Eimeria acervulina (250,000 oocysts per bird) on day 15. Growth performance was evaluated from day 13 to 21, and gut permeability was measured by fluorescein isothiocyanate dextran on day 20. The intestinal lesion, intestinal morphology, and oocysts shedding were determined at the end of the trial. The linear and quadratic orthogonal polynomial contrasts were used to evaluate the effects of increasing NP doses in responses to Eimeria challenge. The results showed that NP was not able to maintain efficient growth performance but improved gut leakage during Eimeria infection period. On the other hand, Eimeria infection increased gut permeability (P < 0.0001) and reduced ileal digestible energy (IDE) and apparent ileal digestibility (AID) of nitrogen. However, the increase of NP linearly enhanced IDE and AID of nitrogen (P < 0.01). Moreover, an interaction between challenge and linear dosage effects was observed for IDE (P = 0.0066) and AID of nitrogen (P = 0.0462). The results indicated that NP improved nutrient digestibility and reduced total oocysts shedding in birds challenged with Eimeria spp. Besides, higher NP doses numerically improved villi height in the intestine. In summary, NP was not able to maintain growth performance of birds but presented positive outcomes on nutrient digestibility and reduced oocysts shedding during mixed Eimeria infection.

The effects of L-Arginine supplementation on growth performance and intestinal health of broiler chickens challenged with Eimeria spp.

This study evaluated the effects of varying levels of L-arginine (Arg) on performance and intestinal health of broilers challenged with Eimeria. Cobb 500 male chicks (n = 720) were randomly distributed in a 5 × 2 factorial arrangement (6 replicates/12 birds). The main factors were Arg levels (1.04, 1.14, 1.24, 1.34, 1.44%) and challenge or non-challenge with Eimeria. At day 12, in the challenge group, each bird received orally 12,500 Eimeria maxima, 12,500 Eimeria tenella, and 62,500 Eimeria acervulina sporulated oocysts. At 5 d postinfection (dpi), intestinal permeability was measured. At 6 and 14 dpi, performance, intestinal histomorphology, nutrient digestibility, tight junction protein (TJP) gene expression, and antioxidant markers were evaluated. Few interactions were found, and when significant, the supplementation of Arg did not counteract the negative effects of Eimeria challenge. Challenge, regardless of Arg level, increased intestinal permeability, although the expression of Claudin-1, a TJP, was upregulated. At 6 dpi, the antioxidant system was impaired by the challenge. Moreover, growth performance, intestinal histomorphology, and nutrient digestibility were negatively affected by challenge at 6 and 14 dpi. Regardless of challenge, from 0 to 14 dpi, birds fed 1.44% showed higher weight gain than 1.04% of Arg, and birds fed 1.34% showed lower feed conversion than 1.04% of Arg. At 5 dpi, intestinal permeability was improved in birds fed 1.34% than 1.04% of Arg. Moreover, 1.34% of Arg upregulated the expression of the TJP Zonula occludens-1 (ZO-1) as compared with 1.24 and 1.44% of Arg at 6 dpi. At 14 dpi, 1.44% of Arg upregulated the expression of ZO-1 and ZO-2 compared with 1.24 and 1.34% of Arg. The nutrient digestibility was quadratically influenced by Arg, whereas the antioxidant markers were unaffected. Thus, the challenge with Eimeria had a negative impact on growth and intestinal health. The dietary supplementation of levels ranging from 1.24 to 1.44% of Arg showed promising results, improving overall growth, intestinal integrity, and morphology in broilers subjected or not to Eimeria challenge.