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Photomicrographs of chicken jejunum sections stained with hematoxylin and eosin. Alcian Blue was used to stain goblet cells. A) Normal histological structure of non-challenge treatment (NC) in jejunum (200X); B) Challenged treatment (CH) (200x) 1. Increase of lamina propria thickness; 2. Inflammatory cell infiltration in the lamina propria; 3 Inflammatory cell infiltration in the epithelium and 4. Presence of oocysts; These alterations contributed to the highest ISI index (P < 0.05) in coccidiosis challenged group at 21 days in comparison to the non-challenged treatment.
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The present study evaluated the effects of coccidiosis on histological parameters and performance of broilers submitted to a mild challenge with Eimeria sp. A total of 132 broilers were randomly divided into two groups with 6 replicates of 11 birds each: negative control (NC) – birds uninfected and challenge (CH) - birds infected by gavage at day-o...
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... NC ISI score 10.8 ± 0.6). There was no significant difference between groups at the other periods. The CH group presented higher ISI total score in comparison to other groups due to higher scores (P < 0.05) caused by inflammatory cell infiltration in the lamina propria and epithelium, increase of lamina propria thickness and presence of oocysts (Fig. ...
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... In the present study, our results showed a reduction in VH, an increase in CD and decrease in the VH/CD ratio in the infested group by Eimeria spp. comparing to non-infested group which is in concordance with previous findings published by several authors [71][72][73][74]. The real mechanism by which coccidian protozoa penetrate the epithelial cells of the intestinal mucosa remains uncertain [75], but it is known that the second-generation schizonts are mainly responsible for changes in villi morphology [76]. ...
The epidemiological survey was carried out to determine the prevalence of eimeriosis in broiler chickens slaughtered depending the season, to determine the different Eimeria species causing the coccidiosis in poultry farms; and to assess the impact of Eimeria parasite on histomorphological structure and oxidative stress parameters of the intestine.
The study was conducted from December 2018 to December 2019 in the province of Bejaia, Algeria. The intestines chickens (n = 366) were obtained immediately after slaughter, each cut into different segments (duodenum, jejunum, ileum, and caecum). Microscopic and parasitological examinations were performed according to standard procedures. Histomorphometric measurements of intestine were obtained using Image J software. Oxidative stress parameters were carried out from intestine tissue.
Eimeria spp. were detected in 73.77% (95% CI 71.20–76.34) of broiler gut samples. The prevalence varied significantly according to the season, with the lowest rates in winter (42.81%, 95% CI 40.35–45.27) and the highest in autumn (97.92%, 95% CI 97.6–99.4). All seven Eimeria species were identified, most commonly E. necatrix (27.70%), E. brunetti (26.47%), and E. tenella (20.96%). The mean lesion score ranged from 1.51 ± 0.05 to 1.79 ± 0.04. Significant differences in VH/CD ratio of intestinal epithelium (P < 0.001) were observed in different intestinal portions of infested broiler chickens compared to non-infested. The mean MDA concentration of intestinal segments was significantly higher in Eimeria species infested broilers compared to non-infested (P < 0.05). The results show at least one difference in CAT, SOD, and ABTS−+ concentrations (P < 0.05) in both chicken’s groups.
Our results revealed that coccidiosis is extremely prevalent in slaughtered broilers, with an abundance of pathogenic Eimeria species. Also, it was concluded that infestation induces tissue structure alterations which coincides with the oxidative damage.
... has substantial impacts on animal production performance. The damage to the intestinal mucosa caused by coccidiosis has a detrimental effect on the broiler's performance [36,37]. The immune response and inflammatory process in the host also cause a reduction in feed intake and an increase in energy demand in affected sites, harming the flock's zootechnical performance [38,39]. ...
The objective of the current investigation was to evaluate several Eimeria challenges and the resulting alterations in intestinal permeability, intestinal morphology, and intestinal lesion scores in broiler chickens. This study included four groups with ten replicate cages per treatment, in which each group received a different treatment dosage of Eimeria, characterizing high, medium-high, and medium-low challenges. Five days after the challenge, intestinal lesions and permeability were assessed. The results showed that the increase in Eimeria challenge led to a considerable decrease in the height of intestinal villosities, in the ratio between villosity size and crypt depth, and in goblet cells. Moreover, after the challenge, there was a considerable increase in intestinal permeability. In conclusion, the medium-low, medium-high, and high-challenge models can be utilized for experimental infection. In the context of clinical studies, it has been observed that the administration of medium-high and high-challenge doses has proven to be adequate. However, it is advisable to utilize a medium-low challenge level to develop a subclinical challenge model for forthcoming investigations that aim to evaluate nutritional recommendations.
... alteration in nutrient distribution might also have a negative impact on laying hen performance (Allen and Fetterer, 2002;Dalloul and Lillehoj, 2005). It has been observed that the inflammation from infection and the corresponding immune response depress the BWG of chickens by up to 41% (Jiang et al., 2010;Belote et al., 2019). ...
An experiment was conducted to investigate the changes in gastrointestinal physiology, including intestinal leakage, immune response, oxidative stress, along with performance traits, of Hy-Line W-36 laying hens following Eimeria infection at peak egg production. A total of 360 laying hens, at 25 wk of age, were assigned randomly to 5 treatment groups, each consisting of 6 replicate cages, including a nonchallenged control group. The other 4 groups were inoculated with graded levels of mixed Eimeria species as high, medium-high, medium-low, and low doses, respectively. The body weight (BW) and body weight gain (BWG) of laying hens were measured from 0 to 14 days postinoculation (DPI). Average daily feed intake (ADFI) and hen-day egg production (HDEP) were measured from 0 to 15 and 0 to 28 DPI, respectively. Gut permeability was measured on 5 DPI, whereas oxidative stress, immune response, and expression of nutrient transporter genes were measured on 6 and 14 DPI. A significant linear reduction in BW and BWG was observed with increased Eimeria inoculation dosage on both 6 and 14 DPI (P < 0.001, P-Lin < 0.0001). An interaction between the Eimeria dosages and DPI was observed for ADFI (P < 0.0001). Feed intake in the challenged groups decreased starting at 4 DPI, with the most significant drop occurring at 7 DPI, which did not recover until 15 DPI. Following the challenge, gastrointestinal physiology shifted toward the host defense against the Eimeria infection by upregulating mRNA expression of tight junction proteins and immune responses while downregulating the expression of key nutrient transporters on 6 and 14 DPI (P < 0.05). An interaction between the Eimeria inoculation dosage and DPI was also observed for daily HDEP (P < 0.0001). Overall, HDEP was lower in the challenged groups compared to the control. Daily HDEP in the challenged groups dropped from 8 DPI and became similar to the control birds only after 24 DPI. Egg production temporarily ceased in most of the laying hens infected with the high and medium-high dosages of Eimeria. In conclusion, Eimeria infection activated the host immune response, negatively affecting the gastrointestinal health, growth performance, and temporarily ceased the egg production of Hy-Line W-36 laying hens when infected at peak production.
... For evaluation of intestinal tissue, 20 villi per bird were evaluated (160 villi/treatment) at 10£ magnification (40£ used to confirm alterations) using the scanned pictures. The "I See Inside" (ISI) microscopy methodology (patent INPI BR 1020150036019) was used to measure histologic alterations on the intestine, and a final intestinal health index was produced (Kraieski et al., 2017;Belote et al., 2018;Belote et al., 2019). In this methodology, a score from 0 to 3 is given to each alteration depending on the extent of the lesion: the greater the lesion, the higher the score. ...
This study tested the effect of distillers dried grains with soluble (DDGS) inclusion in a broiler diet, with or without supplementation of exogenous enzymes, on the microbiota composition, intestinal health, diet digestibility and performance. A total of 288 one-day-old chickens was assigned to 6 treatments (8 replicate of 6 birds each) according to a completely randomized design with a 3 × 2 factorial scheme with 3 DDGS levels (0, 7 and 14%) and 2 inclusions of exogenous enzymes (with or without a multicarbohydrase complex + phytase [MCPC]). The results exhibited that DDGS inclusion up to 14% did not impair broilers performance up to 28 d, however, DDGS-fed animals exhibited significant improvement with the MCPC supplementation. No effects of the enzymes in the ileal digestibility were found at 21 d. DDGS inclusion in the diet affected dry matter and gross energy digestibility. Broilers fed diets with MCPC were found to have less intestinal histological alteration thus better gut health. No effect of DDGS, enzyme or interaction of those were observed for intestinal permeability and in the serum inflammatory biomarker (calprotectin) at 7 and 28 d. The increase of DDGS percentage in the diet reduced the diversity of the ileal microbiota but increased the cecal microbiota diversity. The inclusion of DDGS showed positive effects on microbiota composition due to a reduction of Proteobacteria phylum in the ileum at 28d and a reduction in the presence of Enterococcaceae family in the ileum at 14 and 28d. The inclusion of MCPC complex might promote beneficial changes in the ileal and cecal microbiota due reduce of Proteobacteria, Bacillaceae and Enterobacteriaceae. The supplementation of xylanase, β-glucanase, arabinofuranosidase and phytase to a DDGS diet improves performance and intestinal health allowing the use of these subproduct in the poultry nutrition.
... Gibson-Corley et al. (2013) suggest that scoring methodologies can be successfully used to obtain data from biologic systems (e.g., tissues) in studies aiming group comparisons. These scoring tools can be applied at different levels of tissues examination, including antemortem imaging techniques, postmortem macroscopic examination, and microscopic histopathologic assessments (Eaton 1 et al., 2007;Chapman, 2014;Kraieski et al., 2017;Belote et al., 2018Belote et al., , 2019Belote et al., , 2021Dal Pont et al., 2021). Finally, a scoring system should exhibit 3 fundamental features: 1) to be definable, 2) to be reproducible, and 3) to provide meaningful results (Crissman et al., 2004). ...
... The inflammatory process aiming to eliminate the parasite and results in reduced feed intake, and increases the energy demands on the affected sites, disrupting the zootechnical performance of the flock (Klasing, 2007;Kogut and Klasing, 2009). According to Belote et al. (2019), lesions observed since the second week of life caused by Eimeria spp. impact negatively (P < 0.05) the broilers performance until 28 d of age. ...
... This can be better understood in Figure 5, which showed 2 villi at same cut and almost the same height (A-1,085 mm vs. B-1,090 mm), but, the villus A present normal and healthy structure, while the villus B present higher proliferation of enterocytes and inflammatory cell infiltration, may impair the functionality of the villus. In fact, the proliferation of immature enterocytes along the villi was also observed in other studies (Kraieski et al., 2017;Belote et al., 2018Belote et al., , 2019Sanches et al., 2020). Sanches et al. (2020) characterize this process with the destruction of the lamina propria by inflammation and its invasion by the proliferated immature enterocytes, which stimulates the chronicity of the process. ...
The intestinal wall has on its surface, protrusions called villi that are responsible for the absorption of nutrients. Commonly, these structures have their dimensions measured to related more area surface with better absorption. However, the measurement of these villi neglects the inflammation and the presence of immature cells that increase the surface area but affect negatively the absorption and compromise the animal performance. The measurements of villi/crypt are traditional tools in animal research; however, they may overlook alterations that impact the mucosal functionality. This study aimed to compare the morphometry of the intestinal villi/crypt with the I See Inside (ISI) scoring methodology, exploring their correlation with zootechnical performance. Therefore, broilers were grouped as nonchallenged (NC) and challenged with Eimeria (CH) and jejunum samples were collected at 22 d for histological analysis. The same villi were submitted to the ISI methodology, which is based on the scoring of 8 parameters related to the inflammatory process, and the measurements of villus height (VH), villus width (VW), crypt depth (CD), crypt width (CW), VH:CD ratio and villi absorptive surface (VAS). The CH group presented higher ISI total score, VW, CD, CW and lower VH, VH:CD, and VAS in comparison to the NC group. While the villi/crypt morphometry did not exhibit correlations with performance, the presence of Eimeria oocysts and the ISI total score was positively correlated (P < 0.05) with the feed conversion ratio (FCR), demonstrating a statistical interaction between high ISI scores and worse performance. In conclusion, a larger villus is not related to better intestinal functionality when this enlargement is unleashed by the immune processes occurring inside. The scoring system that evaluates the type of alteration observed has a direct impact on the animal's zootechnical performance which is not observed with the single metric surface evaluation.
... Pieces the intestinal portions (duodenum, jejunum, ileum and caeca) and the liver were collected for histopathology. The samples were sent to the anatomical pathology laboratory of Hassan II's Agronomic and Veterinary Institute for the preparation of histology slides using the hematoxylin and eosin dye, and lesions were scored using the ISI (I See Inside) method (Belote et al., 2019). ...
... The following statistical model was used: Observed values (Yij) = general mean (µ) + dietary treatments as main effect (Ti; I = CON, T1, T2, T3, and T4) + the random error (eij). Rumen fluid pH was analyzed using the SAS procedure [29] with repeated measurements to determine sampling time and interaction between dietary treatments and time points (sampling at 45 and 75 days or before and after feeding at 0 and 3 h per time point). A Duncan multiple range test was used to compare the means of the dietary treatments when the effect of the dietary treatment was considered statistically significant at p < 0.05. ...
... All means were presented with the standard error of the mean (means ± SEM). A Pearson correlation between rumen bacteria metabolites and rumen pH, papilla height, and total papilla surface area was calculated according to Belote et al. [29]. ...
The objective of the present study was to investigate the effects of sprouted barley inclusion level on the growth performance, digestibility, volatile fatty acids, bacterial quantification, and rumen morphology of growing lambs. Five dietary treatments with sprouted barley (0, 25, 50, 75 and 100%) and nine replicates per dietary treatment were performed on forty-five Awassi lambs (90 days old). The average weight gain, intake, and digestibility of dry and organic matter were recorded. The pH, color, volatile fatty acids, bacterial quantification, and rumen histomorphometry were also determined. The results showed that the average dry and organic matter intake in T2 to T4 and the average weight gain in T4 decreased linearly. In contrast, the digestibility of dry and organic matter by sprouted barley (T2 to T4) was higher. The pH values and rumen color were not affected. Concentrations of formic acid, acetic acid, butyric acid, and the ratio of acetic acid to propionic acid were increased, while lactic acid and total volatile fatty acids were lower in all levels of sprouted barley. In addition, lambs fed T4 had a higher quantification of Anaerovibrio Lipolytica, Butyrivibrio Fibrisolvens, and Streptococcus Bovis quantification. Selenomonas Ruminantium was higher in T1, T2, and T4, whereas Megashpaera Elsdenii was lower in T1 to T3. The rumen histomorphometric was improved by sprouted barley (T2 and T3). Sprouted barley improved digestibility and rumen histomorphometry and increased the concentration of some volatile fatty acids and rumen bacteria but resulted in a decrease in average dry and organic matter intake, which negatively affected weight gain in lambs fed 100 % sprouted barley. Further studies are required to determine the potential effects on growing lambs fed sprouted barley.
... All values were expressed as mean ± standard error of the means (SEM). Pearson A correlation between cecal pH and the microbiota was calculated [33]. ...
Gum Arabic (GA) belongs to the Fabaceae family and contains indigestible soluble fibers (80–85%) that could be fermented by commensal bacteria to enhance performance, immune response, and intestinal integrity. This study aimed to investigate the effects of GA on performance, serum biochemical indicators, microbiota, immune-related gene expression, and histological changes in chickens. Six GA levels (0.0, 0.12, 0.25, 0.5, 0.75, 1.0%) were allocated using a total of 432 1-day-old male chickens (12 replicates with 6 chickens each). Growth performance was evaluated on days 10 and 24 of age. Blood parameters, organ pH levels, and intestinal health were determined on day 10 of age. Results showed that GA at 0.12% increased weight gain and 0.12 to 1.0% decreased feed intake but was best in feed conversion ratio and production efficiency except for 1.0% on day 1–10 of age. There was an increase in the thymus weight at GA level 0.25 to 0.75%. GA decreased the pH value of the proventriculus (at 0.50 and 1.0%) as well as the duodenum and cecum (at 0.12 and 1.0%). Chickens fed GA between 0.25 to 1.0% had higher protein and HDL, but lower cholesterol, LDL, and creatinine. Globulin was increased at 0.50% GA, while glucose and triglycerides were decreased (at 0.25 and 0.75% GA, respectively). The immune-related gene expression was reduced, except for 0.25% GA, which increased IL-10. Furthermore, chickens fed GA (0.25 to 0.75%) had higher Lactobacillus spp. and lower Salmonella spp. and Escherichia coli. When chickens received GA, the villus length and length to crypt ratio were higher, which also improved the integrity of intestinal epithelial cells and early duodenal development. We conclude that using GA (0.25 to 0.75%) as a natural prebiotic positively affects the performance, microbiota, immune response, morphology, and gut health of post-hatched chickens. More studies are needed to determine the potential mechanism of GA on broiler chickens.
... To obtain the final value of the ISI index, the IF of each alteration is multiplied by the respective score number, according to the formula ISI = S(IF × S), where IF = impact factor and S = Score. A low ISI score represents better health (Kraieski et al., 2017;Belote et al., 2019). The overall health ISI score considers the following organs/systems and their respective alterations: skin (foot pad lesions and oral mucosa lesions), locomotor (muscular hemorrhage, femoral head necrosis and tibial dyschondroplasia), respiratory (tracheitis, hydropericardium and airsacculitis), liver (color and size), kidney and pancreas (size), proventriculitis, gizzard erosion, yolk persistence, intestine (morphologic alteration on serosa or mucosa layer, cell debris and red thick mucus on mucosa, necrosis, presence of gas, presence of non-digested feed and Eimeria lesions). ...
... These parameters have a great impact on the reduction of an organ's functionality, and unlike the measurement of villi and crypt length, the ISI goes deeper generating more information about the real status of the low-grade inflammation. Therefore, the ISI methodology is an important tool to assess intestinal health and has been used with different types of intestinal challenge models in poultry (Kraieski et al., 2017;Belote et al., 2019;Adewole et al., 2021;Belote et al., 2021). At the macro-intestinal level (sum of the scores given to duodenum, jejunum, ileum, and cecum), the dietary supplementation of P(OA+EO) significantly reduced the ISI score of pullets at week 6 and 21, mainly due to the reduction in the morphologic alteration on serosa or mucosa layers, cell debris, and reddish on mucosa. ...
... Reduced lamina propria thickness, epithelial thickness, epithelial plasma cell infiltration, lamina propria inflammatory infiltration and congestion were the parameters that mainly explained the lowest histologic ISI score of pullets supplemented with P(OA+EO). Belote et al. (2019) reported that lamina propria thickness and inflammatory cell infiltration are the most appropriate parameters to compare feed additives focused in improving intestinal health due to their great correlation with growth performance. In addition, at week 21, the lowest histologic ISI score of pullets on P(OA+EO) was also due to the reduced enterocytes proliferation, goblet cells proliferation and presence of Eimeria. ...
We investigated the dietary supplementation of protected organic acids and essential Oils P(OA+EO) in pullets raised under commercial conditions. A total of 208,000 pullets Lohmann at 2-weeks-old were assigned to receive 1 of 2 treatments: T1, control diet used in the farm; T2, control diet and P(OA+EO) at 300 g/t (fumaric, sorbic, citric and malic acids + thymol, eugenol and vanillin microencapsulated in hydrogenated vegetable fat - Jefo Nutrition Inc. Canada). P(OA+EO) was supplemented from 2 to 18 weeks and the trial lasted 21 weeks. At weeks 6, 12 and 21, 12 pullets/treatment were used for blood sampling and necropsy for ISI – I See Inside . A completely randomized design consisting of 2 treatments, each with 12 replicates of 1 hen/replicate, was used. To evaluate intestinal integrity, birds were inoculated with fluorescein-isothiocyanate labelled dextran (FITC-d) and blood samples were collected after 1.5 h. The macroscopic ISI score of alterations were classified to be presented as: overall health ISI (sum of the scores assigned for intestine, liver, proventriculus, annex glands, locomotor, and respiratory systems) and macro-intestinal ISI (sum of the scores assigned for duodenum, jejunum, ileum, and cecum). The histologic intestinal ISI alterations were evaluated in the ileum. A low ISI index represents better health status. Pullets on P(OA+EO) had lower ( P< 0.001) levels of FITC-d recovered in the blood, which is related to reduced leaky gut. They also presented lower overall health ISI score at weeks 6 ( P = 0.002) and 12 ( P = 0.003), lower macro intestinal ISI score at weeks 6 ( P = 0.0001) and 21 ( P = 0.004) and, lower histologic intestinal ISI score of alterations at weeks 6 ( P = 0.09), 12 ( P = 0.0006), and 21 ( P< 0.0001), which is associated to better overall health. In addition, at week 21, pullets on P(OA+EO) did not present Eimeria oocysts while the control treatment did ( P< 0.0001). In conclusion, the blend of protected organic acids and essential oils evaluated can be used to improve intestinal and overall health status in commercial pullets.
... In contrast, during the recovery phase (8 to 14 days PI), performance was boosted, and higher ADFI and ADG values were recorded in the challenged groups. These results are not in agreement with other studies that have shown that performance parameters are impaired until 21 and 28 days PI [33,34]. However, the current challenge model involving inoculation of an oocyst mixture provided by a high dose of coccidiosis vaccine was able to recreate the impairment of performance parameters the first week PI seen in coccidial infections. ...
Nutritional strategies to improve gut health of broilers are under research. This study investigated the effect of dietary supplementation with sodium butyrate protected by sodium salts of medium-chain fatty acids as a feed additive on broiler gut health. The first experiment was conducted to evaluate the effect of supplementing at 0.5, 1, and 2 kg/t in broilers housed under optimal conditions. Supplementation at 0.5 and 1 kg/t maintained goblet cell counts at 10days of age (p ≤ 0.05), and supplementation at 1 kg/t decreased intraepithelial lymphocyte counts compared to 2 kg/t at 39days (p ≤ 0.10). Abdominal fat pad levels of lauric and myristic acids were gradually increased by supplement dose (p ≤ 0.05). In the second experiment, the feed additive at 1 kg/t was evaluated in coccidiosis-challenged broilers. Experimental treatments were as follows: non-challenged, control-challenged, and supplemented-challenged treatments. Coccidiosis negatively impact performance and modify histomorphometry and microbiota (p ≤ 0.05). The feed additive increased crypt depth at 7days post-inoculation and goblet cell count at 14days post-inoculation (p ≤ 0.05). Further, supplementation interacted with the microbiota modification led by the coccidiosis (p ≤ 0.05). These results suggest that this feed additive could be a useful strategy to reinforce the gut barrier, especially for birds under coccidiosis-challenge treatments.
