THE POTENTIAL EFFICACY OF ALOE VERA GEL AND YUCCA SCHIDIGERAEXTRACT ON GROWTH PERFORMANCE, INTESTINAL LESIONS AND INFLAMMATORY RESPONSE IN BROILER CHICKENS CHALLENGED WITH COCCIDIA

Assiut University web-site: www.aun.edu.eg

THE POTENTIAL EFFICACY OF ALOE VERA GEL AND YUCCA SCHIDIGERAEXTRACT ON GROWTH PERFORMANCE, INTESTINAL LESIONS AND INFLAMMATORY RESPONSE IN BROILER CHICKENS CHALLENGED WITH COCCIDIA

GHADA ALLAM ABD EL –DAYEM¹;REHAB R. ABD ELMAGED²AND NESMA RASHEED³

¹ Poultry Diseases Department, Animal Health Research Institute (AHRI) (Mansoura Branch), Agriculture Research Center (ARC), Dokki, Giza, Egypt.
²Parasitology Department, Animal Health Research Institute (AHRI) (Mansoura Branch),
Agriculture Research Center (ARC), Dokki, Giza, Egypt.
³ Pathology Department, Animal Health Research Institute (AHRI) (Mansoura Branch),
Agriculture Research Center (ARC), Dokki, Giza, Egypt.

Received:28 March 2021;     Accepted:26 April 2021

Corresponding author:GhadaAllamAbd El –Dayem

E-mail address:anasesra@gmail.com

Present address:Poultry Diseases Department, Animal Health Research Institute (AHRI) (Mansoura Branch), Agriculture Research Center (ARC), Dokki, Giza, Egypt.

INTRODUCTION

Avian coccidiosis is one of the most economically important protozoan diseases in the broiler, caused by genus Eimeria.(Noack et al.,2019).E.acervulina,E.maxima and E.tenella are most frequently found in intensively poultry system and the latter is highly pathogenic(Thenmozhi et al.,2014;Clark et al.,2016).Coccidial infectiondamage the epithelial cells of the intestinal mucosal leading to nutrient malabsorption, inefficient feed utilization, poor growth rate,high mortality and secondary bacterial infections (Lee et al., 2012;Huang et al., 2018).Also, it cause economic losses due to increase costs of treatment and prophylaxis. Indeed in a sub-clinical form, it may cause immunosuppression that makes the bird more vulnerable to secondary disease conditions(Akhtar et al.,2012).

Chemoprophylaxis and anticoccidial feed additives have long been used as the main strategy to control avian coccidiosis.Unfortunately, the development of Eimeriastrainsresistance to multiple drugs and residues in poultry products may be potentially hazardous to consumers have made increasing interest for alternative products to control coccidiosis(Cervantes 2015). One of the ways, usingbotanicals in the control of avian coccidiosis, as they are novel natural products containsnew therapeutic molecules to which immunity has notyet developed.

Aloe vera (AV) (Aloe barbadensis Miller) is one of plants, having a great many medicinal and antibiotic properties (Christaki and Florou-Paneri 2010;Kar and Bera 2018).Aloe vera (AV)contains over75bio-logically active compounds include anthraquinones, polysaccharides, vitamins, enzymes and low molecular weight compounds (Choi and Chung 2003) which gives Aloe vera antibacterial, antifungal, antiviral, anti-inflammatory, antioxidant and  immunomodulatory properties (Ahlawat and Khatkar 2011;Radha and Laxmipriya 2015;Akram et al.,2019).Aloe veragel contains polysaccharides, including acemannan which can improve the humoral immune response and cellular immunity(Du et al., 2011).Previous studies explained that  Aloe vera has positive influence of on growth performance, gut microflora, and immune response (Kar and Bera 2018). Also,Aloe veraimproved growth performance,reduced fecal oocyst shedding and lesion score in broilers challenged with coccidia(Yim et al.,2011;Akram et al.,2019;Ahmad et al.,2020).

Yucca schidigera(YS) is a natural, nontoxic product used commercially as a source of steroidal saponins. Moreover, it is a source of many polyphenolic compounds, such as resveratrol and a number of other phytochemicals like yuccaols(Alagawany et al., 2015). These chemical constituents have an antibacterial, anti-inflammatory, antioxidant and immunomodulatory effect(Cheeke et al., 2006).Dietary supplementation of Yucca schidigeraextractimprove nutrient digestibility, growth performance in broilers(Sahoo et al.,2015;Sun et al., 2017).Saponins improve the absorption of nutrients by the intestinal mucosal surface (Barnes et al.,2003). Several studies have shownAnticoccidial  effect of Yucca schidigera extract in broilers  (Djezzaret al.,2014;Bafundo et al.,2020).

Keeping in viewof the facts stated above, the present study was planned to investigate the   beneficial effects ofAloe veragel and Yucca schidigeraextract on growth performance, lesion score, oocyst shedding,histopathological changes and inflammatory responsein broilers experimentally infected with coccidia.

MATERIALS AND METHODS

 Herb extracts and anticoccidial drug:

Aloe vera gel: The plant part (leaves) was identified by a taxonomist in Faculty of Agriculture Mansoura University. The mucilaginous leaf gel was separated from A. veraleaves within 3-4 h post collection to avoid aero deterioration of gel contents according to the method described byLin et al.(2005) and Isah et al. (2019).Gently,the prewashed A. veraleaves were incised longitudinally with the help of a sharp sterilizedknife followed by gentle scraping of gel using aspatula. The gel was homogenized, filteredthrough cheesecloth and stored in screw-cappedjars at 4°C till further use.

Yucca schidigera extract:Yucca extract as a commercial product(SantufoLogestics Corporation,Canda). Each liter containYucca schidigera extract200gm (saponin10gm), Mann oligosaccharides 10gm, citric acid (98%) 10 gm, sodium benzoate 5 gmand purified water up to 1 liter.

Eimeria Challenge:

Field intestinal strains of Eimeriasppwere subjected to isolation, propagation, purification and sporulation as describedby (Tanweer et al.,2014).Briefly, the intestine of the infected birdswere collected. The contents of the gut were collected and soaked overnight in2.5 % potassium dichromate solution. The suspension was filtered and centrifuged at 1500 rpm for 3 min. The supernatant was discarded, and the sediment was resuspended in asaturated solution of sodium chloride and centrifuged at1500 rpm for 3 min. The sediment containing oocysts wasseparated and kept in incubator at 30 °C for 24–72 h. The sporulatedoocysts were stored at 4 °C in potassium dichromate solution, McMaster technique was used for counting of oocyst(Gibbons et al., 2016).Then were inoculated intra crop in broiler chickens in a dose of o 50,000 sporulatedoocysts at 14^th day of age(Munir et al., 2018) except negative control.

Experimental Animals and study groups: A total of 210 one-day old Cobb chicks were divided into equalseven groups, each of 30 birds(3 replicates / group). The birds were fed with coccidiostat free experimental feed. The birds were vaccinated with New Castle Disease (ND) and Infectious Bronchitis (IB) at day 1, Infectious Bursal Disease (IBD) at day 8 and thenND at 14 day.

Group1(G1):non challenged and nontreated.

Group 2(G2):challenged and non treated.

Group 3(G3): challenged and treated  withAloe vera gel 5gm/L from 7^th day of agetill the end of the experiment.

Group 4(G4): challenged and treated withYucca schidigeraextract200mg/Lfrom 7^th day of age till the end of the experiment.

Group 5(G5): challenged and treatedAloe vera gel 5gm/Lfrom 15^th day of age till the end of the experiment.

Group (G6): challenged and treated withYucca schidigera extract 200mg/Lfrom 15^th day of age till the end of the experiment.

Group7 (G7): challenged and treated with Amprolium1g/ 2 L of waterfrom15^th day of age for five successive days.

The broiler chickens were fed ad libitumand they received a lighting regimen of 23 h light: 1 hdarkness. The initial temperature was 32 °C which gradually reduced according to breeding standards. Control parameters, such as temperature, humidity, light and ventilation, were the same for all treatments.

Evaluation of supplementation effectiveness:

Performance parameters:

Feed intake and body weight gain were weekly recorded throughout the study.Feed Conversion ratio were calculated(total feed intake / body weight gain)(Sainsbury 1984).

Anticoccidial evaluation:

At days 7 and 14post infection, three chickens from each group were selected randomly and killed. Intestines along with caeca were removed from the infected birds for the lesion scores (0 to + 4counting system) as described by (Tanweer et al., 2014).The lesions were comprised of petechial hemorrhages, thickening of wall, bloody fecal contents, and mucoid discharge. Based upon the severity of the lesions, a score of 0(no lesions), 1 (mild lesions), 2 (moderate lesions), 3 (severe lesions), or 4 (extremely severe lesions). The bloody diarrhea score for fecal materials as excreted droppingsfrom each group was determined on a scale from 0 to 4 using previouslydescribed methods(Youn and Noh 2001)as well as the  number of oocyst  per gram feces excreted ( OPG) on 3^rd,5^th,7^th,9^thand 11^th days post infection (dpi) by the modified McMaster technique as described by(Alnassan et al., 2013). The reduction in oocyst production (ROP) was calculated with the formula ROP = (oocyst output of positive control group - oocyst output of negative control or medicated groups) × 100/oocyst output of the positive control group.

Inflammatory responses in serum: At 5 and 10 day post challenge three blood samples were collected from each group.Serum sample were centrifuged at 3000 rpm for 5 min. The collected sera weresubjected to determination ofinterleukinIL-1β and IL6 concentrations by using ELISA kits (Becton, Dickinson and Company, Franklin Lakes, NJ) following the manufacturer’s instructions.

Pathological assay:At first week post infection (two weeks post treatment for G3&G4 and one week post treatment for G5,G6&G7)and after 2 weeks post infection(three weeks post treatment for G3&G4 and two weeks post treatment for G5,G6&G7),three bird /group were euthanized by neck severing and necropsied. Samples of about two centimeters from the intestine (duodenum,jejunum and ileum) and cecum were cut, pinned and stowed in 10% buffered formalin solution. The samples were processed by routine histology methods, embedded longitudinallyin paraffin, sectioned 5 µm thick parallel to the cut edge and stained with hematoxylin and eosin (H&E)(Suvarna et al., 2018).

Statistical analysis:

Differences between groups were analyzed by using One-Way ANOVA and Duncan's multiple comparison Post Hoc tests

RESULTS

Growth performance parameters:

Our results showed that there was significant increase(p<0.05) in body weight gain andsignificant decrease (p<0.05) in feed conversion ratio in all challenged treated groups as compared with control positive group(G2).Chickens in groups(G3&G4) administered withAloe vera gel andYucca schidigera extract before infection showed significantly higher (p < 0.05) body weight gains as compared with(G7)treated with amprolium and (G5&G6) administered with Aloe vera gel andYucca schidigera extract after infection.In addition, there was significant difference(p<0.05) in body weight and body weight gain between G3&G4.while, there was no significant difference(p>0.05) in body weight and body weight gain between G5&G6. Birds from the positive control treatment showed highest FCR (P <0.05) compared to all other challenged treatments. There was no significant difference (p>0.05) in FCR between all challenged treated groups(Table 1).

Clinical signs:

In this study, no clinical abnormalities were observed inchickens of the control group (G1), while, chickens in positive control
group (G2) exhibited the typical symptoms of coccidiosis including depression, dullness, ruffled feathers, reduction of feed intake, and blood stained whitish to brownish diarrhea, accompanied with progressive weakness leading to death within 3–5 days after infection. The severity of clinical signs in G7,G3and G4 was less sever than G5&G6.

Bloody diarrhea:

After infection, bloody diarrhea was observed in control positive groups (G2) from 3DPI. and in infected treated groups, started one day later. The severity of bloody diarrhea in all infected treated groups was milder compared with the infected non treated groups.The severity of bloody diarrhea was milder in (G7, G3 &G4)than (G5&G6). Only one chick died from G3,G4&G6, 2 out of G5, while 4 from G2, whereas no deaths were recorded in both G1 and G7as shown in Table(2).

Lesion score:

Regarding to the effect of different treatments on lesion score, our results showed that there was a significant decrease (p<0.05)in intestinal lesion score in all infected treated groups as compared with control positive group (G2).The most significant decrease was observed in G7 and G4 as compared with G3, G5 and G6 (P< 0.05)(table3).

Oocysts output:

The effect of the dietary treatments on fecal oocyst excretion in broiler chickens postinfection is presented in table 4. Fecal oocystexcretion peaked at 5, 6 and 7 dpiand then declined again gradually. The faecaloocyst counts in G3, G4 G5,G6&G7 were significantly (P < 0.05) lower as compared with positive control group(G2). The G7,G4&G3had the lowest number of oocyst count with anROP value of 76.52%,45.31%&39.89% respectively.

Pro inflammatory cytokine serum level:

Our results showed that coccidia infection significantly (p<0.05) up regulate pro inflammatory cytokineIL-1βand IL-6 in all challenged groups as compared to control negative group (G2). Aloe vera gel and Yucca schidigera extract significantly down regulate (P<0.05)expression of pro inflammatory cytokines as compared to control positive group(G2).There was significant decrease(P<0.05) in pro inflammatory cytokines levels in G3,G4 &G7 as compared to G5&G6.(fig1 A&B).

Histopathological finding:

At first week post infection(two weekspost treatment for G3&G4 and one week post treatment for G5,G6&G7) ,intestine in infected untreated group (G2) showed  involvement of the intestinal crypts and glands by the different developmental stages of coccidia,desquamated mucosal epithelia, inflammatory cells (eosinophils& plasma cell) and extravasated erythrocytes in the intestinal lumina (Fig 2A). Epithelial cells were markedly degenerated and necrotic. Ball-Like collections of coccidial developmental stages were seen replacing most of the intestinal glands. Superficial mucosal necrosis and desquamation could be detected (Fig 2B). Large intestine in (G3)showed different developmental stages of coccidiaincluding gametes and oocytes and moderate round cells infiltration, mainly lymphocytes and plasma cells in submucosa (Fig3). The superficial cecal mucosa revealed surface epithelial atrophy and degeneration. While, in G5 large intestine represented severesubmucosal vascular hyperemia, hemorrhages, large number of coccidial developmental stages with partial or complete replacement and destruction of the glandular epithelium (fig4). Cecum in  (G4)showed moderate  involvement of intestinal crypts bydegenerated developmental stages of coccidian with moderate round cells infiltration ( plasma cells) in  submucosa   (Fig5).while, in (G6) large intestine showed developmental stages of coccidia in the intestinal crypts and glands with complete or partial replacements of their epithelial lining(Fig6) and moderate submucosal infiltration by round cells.Large intestine in G7 demarcated failure of the traditional anticoccidial drug from complete relief and healing of the pre-existing coccidial lesions at this time schedule of the experiment with still presence developmental stages, although some were degenerated particularly in the intestinal glands(Fig7).

After 2 weeks post infection(three weeks post treatment for G3&G4 and two weeks post treatment for G5,G6&G7) large intestine of  (G3)  revealed characteristic therapy effect of the used  supplemention, where large number of the still existing coccidial developmental stages were deformed (Fig8). The submucosa showed  mild to moderate infiltration of round cells. The mucosal and submucosal epithelial cells were regenerated and appeared healthy with presence of variable number of goblet cells. While, (G5)showed large number of coccidial oocytes in cecal lumen, some of them appeared deformed or degenerated with absence of  inflammatory or blood exudates. The intestinal crypts and glands showed moderate involvement of their epithelia by developmental stages of coccidia, many of them were degenerated(Fig 9). Examined sections from intestines of( G4) pointed out the most promising and innovated plant extract compounds as nearly most of the intestinal crypts and glands were free from any of the coccidial developmental stages,with marked regenerative changes in the epithelial cells (Fig10). While in (G6)large intestine showed a previously existing remnants from coccidial developmental stages, most of them were degenerated. The mucosal and glandular epithelium appeared regenerated with large hyperchromatic nuclei (Fig 11). Large intestine of G7,most of the intestinal crypts and glands were free from any of the parasitic developmental stages and the previously infested cells exhibited regenerative changes. Mild submucosal infiltration of round cells.(Fig 12).

Table1 :Effectof Aloe vera gel and Yucca schidigera extract on growth performance parameters in broilers chickens experimentally challenged with coccidia.

SEM=Standard Error of Mean. a-e Values followed by different superscript letters were significantly different at (P <0.05).

(G1):non challenged and non treated; (G2):challenged and non treated. (G3): challenged and treated withAloe vera gel 5gm/L from 7^thday of age till the end of the experiment; (G4): challenged and treated withYucca schidigera extract 200mg/L from 7^th day of age till the end of the experiment. (G5): challenged and treatedAloe vera gel 5gm/L 15^th day of age till the end of the experiment. (G6): challenged and treated withYucca schidigera extract 200mg/L from 15^th day of age till the end of the experiment. (G7): challenged and treated with Amprolium1g/ 2 L of waterfrom15^th day of age for five successive days

Table2:Effect of Aloe vera gel and Yucca schidigera extract on severity of bloody diarrhea and number of deaths in broilers chickens experimentally challenged with coccidia

Table3: effect of Aloe vera gel and Yucca schidigera extract on lesion scores   in broilers chickens experimentally challenged with coccidia

Table4: effect of Aloe vera gel and Yucca schidigera extract on oocyst  output  in broilers chickens experimentally challenged with coccidia

Fig.(1): Effect of Aloe vera gel and Yucca schidigera extract on serum level of proinflammatory cytokines in broilers chickens experimentally challenged with coccidia.

DISSCUSION

For long periods anticoccidal drugs are used successfully to ameliorate the adverse effect coccidia infection in poultry. However, alternatives to combat coccidiosis has become of great importancedue to  increase in resistance against anticoccidial drugs (Oelschlager et al.,2019).Phytochemicals are emerging as new alternative methods to control coccidiosis(Kaingu et al., 2017).Keeping in view the wide range of Aloe veraand Yuccaschidigera activities,the present study was conducted to investigate their ability toalleviates the negative impact of Eimeriainfection in broiler chickens.

In the current study, Eimeriachallenge significantly reduced growth performance of infected birds including body weight gain, feed intake, and increase FCR as compared withnon challenged birds. These  results  were in close agreement with previous observation showing that growth performance of broiler chickens impaired by coccidia challenge(Kaingu et al., 2017;Oelschlager et al.,2019). Pathological damage of the intestinal wall during Eimeria infection lead to poor nutrient absorptionwhich, in turn decrease weight gain (Kipper et al., 2013).Regarding to effects of Aloe veragel andYucca schidigeraextract on body weight gain, there was significant increase in body weight gain (p<0.05)in (G3&G4) supplemented with these treatment as prophylactic than G7 treated with amprolium and (G5&G6) supplemented with these treatment as therapeutic. On the other hand,there was a significant difference(P<0.05) in body weight gain between G3&G4 while, there was no significant difference (P>0.05) in body weight gain between G5&G6.There was no significant difference(p>0.05) in FCR between all challenged treated groups.

Similar to the finding of the current study,Alfaro et al.(2007)and Bafundo et al.(2020)reported that dietary supplementation of Yucca schidigera extract(YSE) induced significant increase in bodyweight gain and better FCR in broilers challenged with Eimeria as compared to infected non treated groups.On the contrast,Oelschlager et al. (2019)reported thatYucca schidigeraextract had no significant influence on growth performance include body weight gain and FCR of broilers challenged with Eimeria. The growth-promoting effects of YSE  is attributed to synergistic effect of steroidal saponins and phenolic componentsthat improve nutrient absorption by increasing  intestinal permeability(Alagawany et al., 2016). Also, dietary supplementation with saponins resultin the emulsification of oil fats, promoting their digestion.(Begum et al.,2015).

Isah et al. (2019)reported that supplementation of drinking water with AV gel significantly increase body weight gain in broilers challenged with coccidia. Also,Ahmad et al.(2020)reported that supplemention of drinking water with AV gel (5gm/L) significantly increase body weight  gain and decrease FCR  in broilerschallenged with coccida as compared to control positive groups. On the contrary,Mmereole (2011) reported that dietary inclusion of Aloe veraleaves powder in broilersat 1% has no significant difference in body weight gains as compared to the control group.Growth hormones (auxins and gibberellins) and polysaccharide (glucomannan) components of Aloe vera  able to reduce the inflammation and haemorrhages in theintestines caused by Eimeriainfection that led to better digestion and enhanced weight gain(Surjushe et al., 2008).In addition, AV gel contains several beneficial ingredients including vitamins, minerals, enzymes, organic acids, and carbohydrates which could improve productive traits in broilers(Yim et al., 2011).

Regarding to anticoccidialactivity, our results showed that yucca schidigeraextract and Aloe veragel significantly reduced the intestinal lesion as compared to positive control group.  Among treated groups (G7) and (G4) had less severe lesion score as compared with (G3), (G5) and (G6). These observations agree with those described by Ahmad et al.(2020) who  reported that Aloe veragel reduced intestinal lesions in broilers chickens challenged with coccidia as compared to positive control group. Likewise,Bafundo et al.(2020)reported that dietary yucca schidigeraextract reduced intestinal lesion in broilers chickens challenged with coccidia.

Concerning the fecal oocyst output, yucca schidigera and aloe veragel supplementation significantly lower oocyst per gram(OPG) as compared with positive control group.yet, it failed to do as the infected medicated group with amprolium (G7).The highest reduction in(OPG) was (76.25%,45.31%&39%) in G7,G4&G3 respectively. While, G5&G6 had the lowest reduction inOPG(34.3%and32.43%). The reduction in OPG is an indicative of a lower degree of infection (Kucukyilmazet al., 2012).The reduction in oocyst count is an evidence of improvement bird immunity to resist coccidian infection. (leeet al.,2013).

These findings  corresponded  with Alfaro et al.(2007)andBafundo et al.(2020) who  reported that dietary Yucca schidigerareduced oocyst output in broilers challenged with coccidia.The anticoccidial activity of Yucca schidigeramight be ascribed to saponins content of Yucca schidigera that inhibit the development of protozoa by interacting with the cholesterol present in the parasite cell membrane, resulting in parasite death(Francis et al., 2002).Also, It was assumed that saponin did not destroy the oocyst’s wall but entered the wallthrough the micropyle cap or gap and directly disturbed the sporocyst(Wina 2018).

In addition,(Yim et al.,2011;Kaingu et al., 2017;Ahmad et al., 2020) reported that dietary supplementation ofAloevera significantly reduced mean of OPG in coccidia challenged broilers. Aloe veramay interfere with critical stages of Emeria development and reduces damages to the intestinal wall of the chicken by reducing oocysts count, lesion score and haemorrhage(Yim et al.,2011;Kaingu et al.,2017;Ahmad et al., 2020).The polysaccharide derivatives, in Aloevera gel, has antisporulation effect by interfering in the physiological processnecessary for sporulation process like preventing access ofoxygen and inhibition of various enzymes responsible for sporulation (Khan 2012).The chemical constituents in Aloe extract as 1, 8 dihydroxyanthraquinone and its derivatives include Aloeemodin, aloetic acid and isobarbaloin act as laxative agents that increase bowel motility. This leads to the quick discharge of coccidialoocysts that are lodged in faecal matter thereby reducing the oocyst count (Nghonjuyi et al., 2015).

Cocidiosis is well known to result in strong intestinal inflammation that is partly mediated by an overexpression of cytokines (Lillehoj et al. 2004).In the current study,Eimeria challenge induced significant increase(p<0.05)in IL6& IL-1β level in serum of positive control group as compared to other treated groups.Our findings corroborate previous reports increase level of IL-6, IL-1β due toEimeriainfection in chickens,(Hong et al., 2006;Grenier et al.,2016;Moraes et al., 2019). While, supplementation of Yucca schidigeraand Aloe veradownregulatelevels of IL6& IL-1β. These finding indicate that these supplements exerted an anti-inflammatory effect.Oelschlager et al.(2019) reported thatYucca schidigeraextract ameliorated the upregulation of IL-1β level in broiler experimentally challenged with coocidia.Saponin supplementationmay possess some measurable immunomodulatoryeffects during Eimeriainfection.YSE is a rich source of polyphenols such as resveratrol(Piacente et al., 2005). Resveratrol could inhibit the NF-jB pathway that result in the decreasetranscription of proinflammatory cytokines like IL-1βand TNF-α.(Zhang et al., 2014).Acemannan, a major polysaccharide present in AV gel, appeared to be able to binds to macrophage receptors and stimulates the synthesis of cytokines (interleukin 1 (IL-1), interleukin 6 (IL-6)) and tumor necrosis factor-alpha (TNF-α) (Djeraba and Quere 2000.;Darabighane et al., 2012).

The severity of lesions and the number of parasites observed at histologicalpathological examination are in accordance with the recordings for the macroscopic lesion score and the oocysts counts.At first week after infection, all challenged treated groups showed less inflammatory and degenerative changes and reduced coccidial developmental stages as compared to control positive groups. Among treated groups, the most improvement in inflammatory changes and reduction in coccidial developmental stages was observe in (G7) treated with amprolium followed by(G 4) treated with yucca and (G3) treated with aloe vera as prophylactic Similarly, after two weeks post infection (G7) treated with amprolium showed the most improvement where the intestinal crypts and glands were free from any of the parasitic elements and the previously infested cells exhibited regenerative changes.(G4) treated with yucca as prophylactic showed improvement similar to (G7) treated with amprolim where the large intestine appeared free from coccidial stages with marked regenerative changes in crypts and glandular epithelium and absence of inflammatory or hemorrhagic complicating changes of avian coccidiosis. These finding indicates that these supplementation is able to kill or inhibit the growth and development of oocysts,regenerate the damaged cecal tissues and the decrease in inflammatory cells.

Oelschlager et al.(2019) reported that saponin supplementation modified morphological parameters of the jejunum on 14^th day following Eimeria challenge by reducing mucosal thickness. Saponin supplementation may promote regeneration and reconstitution of the intestinal mucosal layer back to normal levels as evidenced by saponin treated birds not being significantly different from the unchallenged birds.

Previous studies reported increasedLactobacillus count and reduced Escherichia coli count inthe gut of broilers by supplementing diets with AV(Sujatha et al.,2017).Short-chain fattyacids, as the final product of fermentation by Lactobacillusbacteria, improve intestinal morphology and might have stimulated the proliferationof epithelial cells of the bowel(Olnood et al., 2015).In addition, lowercrypt depth with Aloeverasupplementation indicated for slow tissue turnover preventing the pathogens from tissue destruction in the gut(Ghazanfari et al., 2015).

CONCLUSION

In conclusion, the results of the present study demonstrated that Yucca schidigera extract and Aloe veragel supplementation has great potentials for improving growth performance, intestinal lesionsand inflammatory response in broilers under coccidia challenge.Therefore, Yucca schidigera extract and Aloe veragel may be a potential and valuable candidate to be used as a low cost alternative drugs to control coccidiosis in chickens. Further researches should be done to investigatethe therapeutic effect and mechanism of action of Yucca schidigera extract and Aloe veragel against coccidiosis in broiler chickens.

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