Impacts of Microbial based Therapy on Growth Performance, Intestinal Health, Carcass Traits and Economic Efficiency of Clostridium perfringens‐Infected Cobb and Arbor Acres Broilers

Abstract The poultry farms need a safe and effective alternative for antibiotics that can counteract the negative impacts of necrotic enteritis (NE), which causes severe mortalities and economic losses. The current study was aimed to examine the influence of antibiotic (Flagymox) and the microbial‐based administration on carcass traits in Clostridium (C.) perfringens‐infected Cobb and Arbor Acres broilers. A total number of 360 Cobb and Arbor Acres broiler chicks (180 numbers per breed) were allocated to four groups; negative control group (without any treatments); positive control group (administration of C. perfringens at the rate of 1 × 109cfu/bird via crop gavage twice daily from day 16 to 18 post‐hatch); C. perfringens challenge plus antibiotic (Flagymox®) group, and Clostridium perfringens challenge plus microbial‐based treatment (Big‐lactoα®) group. The results indicated that the Flagymox and Big‐lactoα treated Cobb breed group achieved a significant increase in their body weight (BW) than the positive control group at the third week post‐infection. Also, the Arbor Acres breed gained significantly higher weight compared to the Cobb breed at the third week. Total weight gain (TWG) from 0 to the fifth week in the Cobb and Arbor Acres breeds were higher in the groups treated with Flagymox and Big‐lactoα compared to the birds challenged with C. perfringens without any treatment, thus, increasing the total return (TR) in the treated groups. Economic efficiency showed no significant differences (p < .05) between the treatment groups of both the breeds. Although the treatment cost of Flagymox is higher than the microbial‐based treatment (0.86 versus 0.35 LE), there were no mortalities reported in the microbial‐based groups in both the breeds resulting in significantly low losses compared to the Flagymox treated groups. The groups treated with the microbial‐based products in both breeds were superior in dressing percentage (75.16 and 77.06% for Cobb and Arbor Acres, respectively) compared to that of the other groups. In conclusion, microbial‐based therapy improved the growth rate, carcass traits, survival rate, and economic efficiency in necrotic enteritis induced in Cobb and Arbor Acres broilers.

of 360 Cobb and Arbor Acres broiler chicks (180 numbers per breed) were allocated to four groups; negative control group (without any treatments); positive control group (administration of C. perfringens at the rate of 1 × 10 9 cfu/bird via crop gavage twice daily from day 16 to 18 post-hatch); C. perfringens challenge plus antibiotic (Flagymox ® ) group, and Clostridium perfringens challenge plus microbial-based treatment (Big-lactoα ® ) group. The results indicated that the Flagymox and Big-lactoα treated Cobb breed group achieved a significant increase in their body weight (BW) than the positive control group at the third week post-infection. Also, the Arbor Acres breed gained significantly higher weight compared to the Cobb breed at the third week. Total weight gain (TWG) from 0 to the fifth week in the Cobb and Arbor Acres breeds were higher in the groups treated with Flagymox and Big-lactoα compared to the birds challenged with C. perfringens without any treatment, thus, increasing the total return (TR) in the treated groups. Economic efficiency showed no significant differences (p < .05) between the treatment groups of both the breeds. Although the treatment cost of Flagymox is higher than the microbial-based treatment (0.86 versus 0.35 LE), there were no mortalities reported in the microbial-based groups in both the breeds resulting in significantly low losses compared to the Flagymox treated groups. The groups treated with the microbial-based products in both breeds were superior in dressing percentage (75.16 and 77.06% for Cobb and Arbor Acres, respectively) compared to that of the other groups. In conclusion, microbial-based therapy improved the growth rate, carcass traits, survival rate, and economic efficiency in necrotic enteritis induced in Cobb and Arbor Acres broilers.

| INTRODUC TI ON
Necrotic enteritis (NE) is one of the most widespread enteric diseases found in broilers. It has a major impact on the cost-efficiency of meat production due to the increased mortality, veterinary intervention, and treatment costs (Whelan et al., 2019). The NErelated global economic losses in the poultry industry are estimated to be over 6 billion US dollars annually (Wade et al., 2015).
This disease causes a multi-factorial gut health issue that depends on interactions between several factors, such as host, infectivity, nutritional and managerial factors. The presence of Clostridium (C.) perfringens, usually found in the healthy chicken intestines, with some of the additional and predisposing factors that modify the intestinal ecosystem allowing C. perfringens to overgrow, eliciting the clinical signs and lesions of NE. Studies (Jang et al., 2013;Tsiouris, 2016) have reported susceptibility differences in Cobb, Ross, and Hubbard broilers in developing NE. Cobb chicken fed with a high protein diet was the most susceptible breed to orally infected Eimeria maxima and viable C. perfringens (Jang et al., 2013;Tsiouris, 2016). Moreover, Kim et al. (Kim et al., 2014) documented the disparate NE susceptibility in two inbred Ross chicken lines having variations in transcriptional profiles of host-pathogen response against the disease with differentially regulated immune genes (Osman & Elhariri, 2013). Several strategies were applied to reduce the incidence of NE and maintain the profitability of broiler production . The usage of antimicrobial drugs, anticoccidials, and/or antibiotics in feed and/or drinking water are increased nowadays to enhance the growth and feed efficiency through their antibacterial activity against C. perfringens (Williams, 2005). A sense of urgency to find an alternative approach has overcome the strong challenges in the global poultry industry as there is an increase in the incidence of NE in Europe and the USA because of the European Union ban in 2006 that restricts the usage of all antibiotics in all animal feed (M'Sadeq et al., 2015). This is also accompanied by the high acquisition resistance of C.
Live microorganisms, prebiotics, plants and their extracts, organic acids, enzymes, lysozyme, yeast extract and antimicrobial peptides are all employed in the intervention strategies used for C.
perfringens induced NE in broiler chickens, which could support a one health approach (Caly et al., 2015). As known, bacteriophages are highly species-specific viruses, and predators of bacteria and hence are widely used to treat bacterial infections in humans, animals and poultry (Caly et al., 2015;Miller et al., 2010;Seal et al., 2018). Some papers have documented the efficiency of using feed supplementation of six bacteriophages to reduce the mortality and improve weight gain (WG) and feed conversion rate (FCR) in the NE challenged chicken. Moreover, probiotics are the live yeast or bacteria used in the feed to ameliorate the adverse effects of the C. perfringens infected broilers on their performance and intestinal health by reducing the risk of opportunistic and pathogenic bacterial infection (Hussein, Ahmed, Abudabos, Aljumaah, et al., 2020;Khalique et al., 2020;Whelan et al., 2019). Therefore, this study was aimed to determine and outline the influence of antibiotics and microbial-based administration (Flagymox and Big-lactoα) on the growth performance traits, intestinal health measurement, carcass traits and the measurement of economic efficiency of C. perfringens infected Cobb and Arbor Acres broilers.

| Measurement of the Growth Performance
During the experiment (days 0-35), the growth performance of the broiler chicks was evaluated by recording their daily and weekly feed intake (FI). The live body weight (BW) was monitored weekly to calculate body weight gain (WG). The feed conversion ratio (FCR) was calculated as reported by Elbayoumi et al., 2014. Also, dead birds in each group were recorded daily to calculate the mortality rate as described by Miller et al., 2010, and European Broiler Index (EBI) was estimated as described by Mathis et al., 2016.

| Assessment of the intestinal health
On day 21 of post-hatch, five birds were randomly selected from each group and humanely sacrificed by cervical dislocation. The intestinal tissues were then collected for the following parameters:

| Bacterial Quantification
To estimate the counts of E. coli and C. perfringens, a piece of the jejunum (15 cm) from the sacrificed birds were placed in 10 ml of anaerobic fluid thioglycollate broth (Oxoid microbiology product, Thermo Scientific Co, USA) under septic conditions for 30 s. 0.5-mL of this sample was placed into 4.5 ml of neutral physiological buffer saline to perform 10-fold serial dilutions, where each dilution was plated onto the MacConkey agar and incubated at 37°C for 24 hr. Also, another 0.5-mL of the sample was used for 10fold serial dilutions in fluid thioglycollate broth, and 1 ml of each dilution was poured on tryptose sulfite cycloserine agar plates and further incubated anaerobically at 37°C for 24 hr. The inoculated plates that had more than 30 or less than 300 colonies of E. coli and C. perfringens with typical morphology were recorded (McReynolds et al., 2004).

| Lesion scoring and Morphometric analysis
To evaluate the gross lesions of necrotic enteritis, 10 cm of jejunum section from the sacrificed birds were examined macroscopically and scored from 0 to 4 using previously documented criteria (Prescott, 1979). Then, the jejunum samples were fixed in phosphate-buffered formalin for at least 24h and embedded in paraffin. Sections of 5 mm were cut and stained with haematoxylin and eosin (Bancroft & Gamble, 2008). The stained sections were microscopically examined for histopathological changes such as epithelial cell necrosis, inflammatory cells, fibrin exudate in the gut lumen, and epithelial hyperplasia, which were recorded and scored between 0 to 4 based on the published papers (Dahiya et al., 2007). Also, the intestinal morphometric variables, including the villi length, crypt depth, villi depth and the extent of tissue injury, were measured using the Image capture. Furthermore, the analysis system (Langhout, 1998;Wealleans et al., 2017) was used to calculate the villi length and crypt depth ratio.

| Total RNA extraction, cDNA synthesis and quantitative real-time PCR (qRT-PCR)
To achieve a better understanding of the effects of NE on jejunal cytokine and chemokine production (IL-6 and IL-8), the effects of antibiotics and microbial-based product administration were examined for their expression in the jejunum. The expressions of jejunal IL-6 and IL-8 genes were analysed by real-time PCR using sense and antisense primers.
Intestinal tissue (jejunum region) was cut longitudinally, washed with saline, blotted on a filter paper and maintained at −80°C until RNA extraction. Total RNA from jejunum was isolated using TRIzol (Invitrogen™, Thermo fisher scientific) reagent as described according to the manufacturer's protocol. The concentration and purity of total RNA were estimated by measuring the optical density at 260 and 280 nm. 5 µg of total RNA from each sample was reverse transcribed into cDNA using  The real-time PCR cycling program was as follows: 95°C for 10 min (holding stage), 95°C for 15 s for 40 cycles (denaturation stage), followed by 60°C for 1 min (annealing and extension stage).
The changes in the gene expression were calculated by the 2 −ΔΔCt method using the cycle threshold (Ct) values, where Δ Ct indicated the Ct changes in target genes compared to a reference (housekeeping) gene (GAPDH) (Schmittgen & Livak, 2008).

| Measurement of carcass quality
At the end of the growing period (day 35), five birds were randomly collected from each group and left for 12 hr to fast. Each bird's carcass was weighed before the slaughter and after complete dressing (removal of the feather, head, neck, shanks, feet and viscera). The dressing percentage was calculated according to the method of Brake et al., 1995. All the internal organs (Heart, gizzard, liver without the gall bladder, spleen, thymus, bursa, abdominal fat and gizzard fat) were also weighed immediately after the sacrifice, and the gizzard was weighed after the removal of its content.

| The measurement of economic efficiency
The economic efficiency at the end of the experiment (day 35) was evaluated using total costs (TC), which included total variable cost (TVC) and total fixed cost (TFC). TVC included feed consumption, total veterinary management (TVM), labour, chick price, water, electricity and litter costs and was estimated in Egyptian pound LE (1 USD ≈ 17.6 LE) for every bird in all the groups during the period of the experiment. TFC was estimated at 1.61 LE/bird, which included depreciation of buildings and equipment (Amarapurkar et al., 2014).

| Statistical analysis
Data obtained in the current study were statistically analysed for analysis of variance (ANOVA) (Significance at p ≤ .05), and the breed effect was analysed by independent sample T-test using SPSS/ PC + "version 16 (Guide, 2002). The results were represented as means ± standard error (SE). Differences between the groups were TA B L E 2 Effect of antibiotic and microbial based administration in Cobb and Arbor acres broilers experimentally infected with Cl. perfringens on body weight changes (BW) and feed intake (FI) analysed using Duncan's multiple Post Hoc tests, and Pearson's correlation was used to analyse the association between the macroscopic lesion scores of the jejunum and its morphometric variables. Table 2  The FI in the fourth and fifth weeks, along with the total feed intake (TFI), showed non-significant differences among different groups.

| The effect of antibiotics and microbial-based administration on body weight gain changes (WG) and feed conversion ratio (FCR) in experimentally infected Cobb and Arbor Acres broilers infected with C. perfringens
The results represented in Table 3 showed that WG at the sec- There was a non-significant difference in the WG between different groups in the fifth week for both the breeds. As for the total weight gain (TWG) from zero to the fifth week in the Cobb breed, a numerical increase was observed in the groups treated with antibiotics and microbial-based products (IC, 1662.

| The effect of antibiotics and microbial-based administration on European broiler index (EBI) in experimentally infected Cobb and Arbor Acres broilers with C. perfringens
The groups treated with antibiotics and microbial-based

| The effect of antibiotics and microbial-based administration on post-infection mortality and livability percentage of experimentally infected Cobb and Arbor Acres broilers with C. perfringens
Regarding the mortality percentage, a difference was observed between the treated and untreated groups of NE infected broilers F I G U R E 1 The effect of antibiotics and microbial-based administration on the European broiler index in Cobb and Arbor Acres broilers experimentally infected with C. perfringens TA B L E 3 Effect of antibiotic and microbial based administration in Cobb and Arbor acres broilers experimentally infected with Cl. perfringens on body weight gain changes (WG) and feed conversion ratio (FCR) Note: Small letters are significantly different between treated and untreated groups (p ≤ .05), while Capital letters significantly differ among breeds. BWG) was calculated by subtracting the body weight between two successive weights in different weeks of experiment, TWG (final body weight-initial body weight) and feed conversion rate (FCR)= feed intake (FI)/ body weight gain (BWG).
( Figure 2). The highest mortality percentages were 8.9% in the NE infected Cobb birds followed by 6.7% in the infected Arbor

| The effect of antibiotics and microbialbased therapy on the counts of C. perfringens and E. coli in the jejunum of experimentally infected Cobb and Arbor Acres broilers with C. perfringens
The diversity of C. perfringens and E. coli in the jejunum of the experimental birds are presented in Figure 3 and  and microbial-based therapies produced a significant reduction

| The effects of the antibiotics and microbial therapy on the jejunal histomorphometric measurements of the C. perfringens infected Cobb and Arbor Acres broiler chicks
The jejunal histomorphometric measurements (villus length, villus width, crypt depth, the ratio between villus length and crypt depth (VH: CD ratio) and the extent of tissue injury) are presented in Table 4. In all the treated and untreated groups, a significant  (Table 4).

| The effect of antibiotics and microbial-based administration on mRNA expression of jejunal IL6 and IL8 in Cobb and Arbor Acres broilers experimentally infected with C. perfringens
Changes in the intestinal (jejunal) IL6 and IL8 mRNA expression are presented in Figure 6. The expression of IL6 and IL8 showed nonsignificant (p < .05) changes in the positive control groups (IB & IIB) compared to other groups with both Cobb broiler and Arbor Acres breeds. The antibiotic-treated groups (IC & IIC) showed up-regulation of IL6 and IL8 mRNA numerically compared to the negative control groups (IA & IIA) either in Arbor Acres and Cobb broiler breeds. In the groups treated with Flagymox and microbial-based products, a numerical nonsignificant (p < .05) normalization was observed in IL6 and IL8 expression compared to the groups challenged with C. perfringens ( Figure 6). Table 5 shows that the groups treated with the microbial-based prod-  However, no losses were observed in the treatment groups of the microbial-based product and antibiotics in both the breeds (ID & IID) and Cobb breed (IC), respectively. Thus, treatment either with antibiotics or with microbial-based products resulted in better results.

| The effect of antibiotics and microbialbased administration on different return parameters (Selling of litter, Selling of broiler chicken, total returns and net profit) in Cobb and Arbor Acres broilers experimentally infected with C. perfringens
The results presented in Table 7 showed that returns from selling broiler chicken, and the total returns (TR) were higher in challenged and treated groups compared to the positive control group of both TA B L E 6 Effect antibiotic and microbial based administration in Cobb and Arbor acres broilers experimentally infected with Cl. perfringens on different cost patterns (total veterinary management, total variable cost, total fixed cost and total cost)  Net profit was higher in all the treated groups of Cobb breed (LE 6.87 and 5.14 for IC & ID) compared to the positive group (IB), while the group with Arbor Acres breed treated with the microbial based product (IID) showed a higher value (LE 6.46) compared to the positive control group (IIB) (LE 5.45). Considering the breed effect, the Arbor Acres breed showed a higher net profit compared to the Cobb breed (LE 6.01 and 4.75, respectively).

| D ISCUSS I ON
Intestinal health is still one of the most important subjects in poultry production because the impact of digestion and nutrient absorption can harm the feed efficiency, increase the susceptibility to diseases, and eventually cause economic losses. Among the intestinal diseases in poultry, NE is economically more important as it impairs production and economic performance in broiler chicken (Caly et al., 2015).
This disease is being controlled by antimicrobials for many years in the poultry industry. Recently, microbial-based products are being used as alternatives in disease prevention that also act as supplements for growth enhancement, aiming to reduce the use of antibiotics in the broiler industry (Seal et al., 2018). Thus, there is an urgent need to explore the full practical potential of microbial therapy. In this context, the present study explored microbial-based product as an alternative for the antibiotics in the treatment of NE in two different broiler breeds, where its effects were investigated on broiler production and economic performance, microbial counts, intestinal macroscopic and microscopic lesion with its histomorphology, carcass composition and gene expression.
Current findings confirmed that the antibiotics treated groups in both Cobb and Arbor Acres breeds achieved an increase in BW, FI and dressing percentage compared to the positive control group in the third week. Similar findings by Abudabos et al. (Abudabos et al., 2018;Abudabos & Yehia, 2013) reported that the C. perfringens challenged group treated by avilamycin had higher body weight, TA B L E 7 Effect of antibiotic and microbial based administration in Cobb and Arbor acres broilers experimentally infected with Cl. perfringens on different return parameters (Selling of litter, selling of broiler chicken, total return and net profit)  supplemented with six bacteriophages (Miller et al., 2010) or treated with probiotics or antibiotics (Hussein, Ahmed, Abudabos, Aljumaah, et al., 2020;Jayaraman et al., 2013). Moreover, the synergistic effect of a combined treatment that includes phages and bacteriocins to control C. perfringens showed a significant reduction in the bacterial population (Heo et al., 2018). Hence, a significant reduction in the mortality rate and intestinal microbes may be due to the inhibitory effects of the antibiotics and microbes towards the enteric microorganisms (Chichlowski et al., 2007).

Items
In the present study, the pathological findings in the jejunum of the infected Cobb and Arbor Acres groups were characterized by the development of NE lesions at day 21 post-hatch. Moreover, the jejunal morphology measurements demonstrated a significant decrease in the villus length and crypt depth, which negatively correlated with the significant increase in the extent of tissue injury compared to the negative control groups of Cobb and Arbor Acres breed. These findings coincide with the previously reported papers that have illustrated the negative effects of C. perfringens and its toxins on the sloughing and chronic inflammation of the intestine, which is also associated with high intestinal lesion scores and shortening of the villus length. This directly affects the nutrient absorption while indirectly affecting the performance of the birds (Gholamiandehkordi et al., 2007;Hussein, Ahmed, Abudabos, Aljumaah, et al., 2020;M'Sadeq et al., 2015;McReynolds et al., 2009;Star et al., 2009;Xu et al., 2003).
The severity of the pathological findings and the histomorphology changes in the jejunum of the infected Cobb and Arbor Acres groups were reduced because of the antibiotics or microbial-based F I G U R E 7 The effect of antibiotics and microbial-based administration on the economic efficiency of Cobb and Arbor Acres broilers experimentally infected with C. perfringens F I G U R E 8 The effect of antibiotics and microbial-based administration on the relative economic efficiency of Cobb and Arbor Acres broilers experimentally infected with C. perfringens treatments. These findings partially concurred with the findings of the other researchers (Calik et al., 2019;Hussein, Ahmed, Abudabos, Aljumaah, et al., 2020;Jayaraman et al., 2013;M'Sadeq et al., 2015) who have reported the ameliorative effects of different feed supplementations such as antibiotics, phytobiotics, prebiotics or probiotics in developing the NE lesions in the intestine of the broiler chickens. This effect may be attributed to the activity of the used drug that reduces the irritation caused by C. perfringens while preserving the integrity of the intestinal epithelial layer. Antibiotics and bacteriophages directly affect the pathogenic bacteria by causing their death and preventing further bacterial growth. Moreover, bacteriophages and probiotics produce anticlostridial factors that maintain favourable ecological homeostasis for the intestinal microbes to promote the development of gut immunity and histomorphology (Caly et al., 2015;Khalique et al., 2020;Li et al., 2018;Seal et al., 2018;Wernicki et al., 2017;Whelan et al., 2019;Zimmer et al., 2002). The results obtained in this study, including no mortalities, low C. perfringens log10 values, mild macroscopic and microscopic NE lesions were consistent with those reported in controlling the C. perfringens infection in the broilers using probiotics (Calik et al., 2019;Hussein, Ahmed, Abudabos, Aljumaah, et al., 2020;Jayaraman et al., 2013;M'Sadeq et al., 2015;McReynolds et al., 2009;Wang et al., 2017) and bacteriophages (Miller et al., 2010). These results occur more frequently than expected because of the synergistic ameliorative effects of the probiotic and bacteriophages used along with the drugs against the C. perfringens infection in the broilers.
IL-6 is a multifunctional cytokine with a pro-inflammatory activity that induces acute-phase protein synthesis as well as aids the adaptive immune response (Kaiser et al., 2000). IL-8 is a chemokine produced by the macrophages that functions primarily as a chemoattractant and plays an important role in inflammation (Shahzad et al., 2010). Here, C. perfringens challenged groups showed a numerical up-regulation in the expression of IL6 and IL8 due to the inflammation induced by the bacterial infection. It is shown that necrotic enteritis in broiler chicken results in increased expression of IL8, while the expression of IL6 is not altered (Park et al., 2008). Challenging with C. perfringens up-regulates the mRNA expression of interleukins (IL6 and IL8) in primary intestinal epithelial cells (Guo et al., 2015). In our study, the group treated with antibiotics showed a non-significant down-regulation in IL6 and IL8 compared to the negative control group either in Arbor Acres or Cobb broiler breeds. The decrease in the expression of the interleukins clarifies the role of antibiotics in treating intestinal inflammation. Both the breeds treated with the microbialbased product showed an up-regulation in IL6 and IL8 expression compared to the antibiotics treated groups, which may be due to the activation of the immune system since the microbial-based product increases the phagocytosis of bacteria by macrophages (Adhikari et al., 2017).
In this study, no significant differences were observed between non-infected and infected birds in the carcass quality, including dressing, bursa, spleen and gizzard percentage, which also coincided with other research findings . Contrastingly, some studies (Elmenawey et al., 2019) have recorded a significant increase in the above-mentioned carcass qualities. In general, the positive control group of each breed has a higher value of intestine percentage compared to the other groups of the same breed, which may reflect the degree of inflammation in the jejunum along with the accumulation of the exudates in the intestines of the infected groups (Abudabos et al., 2016;Alzawqari et al., 2016).
The microbial-based treatment was less expensive (LE 0.35) compared to the antibiotics treatment. Hence, the microbial-based treatment offered an inexpensive treatment. The total returns (TR) were higher in all the treated groups compared to the positive control group of both the breeds. The highest value of TR was found in the antibiotics treated group of Cobb breed, whereas the lowest value was found in the positive group of Cobb breed. Net profit was higher in all the treated groups (IC, ID) compared to the positive control group of Cobb breed. Our results can explain the reported reduction in weight gain, poor performance and increased costs for the poultry producers (Abudabos & Yehia, 2013;Wade et al., 2015). Similarly, necrotic enteritis was considered to reduce the body weight of the infected birds compared to the healthy birds .
Regarding the total losses due to mortality, the positive group recorded the highest losses among different groups of both breeds, whereas no losses were found in the groups treated with the microbial-based product and antibiotics in both the breeds and Cobb breed, respectively. These results were in agreement with the other studies (Geier et al., 2010;Hussein, Ahmed, Abudabos, Aljumaah, et al., 2020;Miller et al., 2010).
Although a non-significant difference was observed in the economic efficiency and relative economic efficiency, the microbial-based product and antibiotics succeeded in improving the economic efficiency (Choct & Kocher, 2008;Skinner et al., 2010;Wade et al., 2015).
It was found that necrotic enteritis reduced the body weight of the birds and increased the cost of production, which could be controlled using antibiotics (Williams, 2005). Also, using alternative strategies to control NE could limit the economic impact of the disease (Dahiya et al., 2007;M'Sadeq et al., 2015;Miller et al., 2010). Moreover, the addition of probiotics had a positive effect on the performance of broiler chickens infected with NE (Hussein, Ahmed, Abudabos, Aljumaah, et al., 2020), which was also reflected in economic efficiency.

| CON CLUS ION
The poultry constitutes 8% of the reported food-borne disease outbreaks, and since the microbial-based treatment prevents foodborne pathogens in the broiler food chain, it confirms the hygienic importance of using the microbial-based treatment in controlling C.
perfringens. Also, the bio-control of C. perfringens infection using the microbial-based product is considered a natural, hygienic and relatively inexpensive method to control poultry health problems. The microbial-based product also protects chickens from being biohazardous because of C. perfringens infection.

ACK N OWLED G EM ENTS
We thank Taif University for financial support for Project number TURSP-2020/09 for Mohamed Mohamed Soliman.

CO N FLI C T O F I NTE R E S T
Authors declare that there is no conflict of interest for current data.

PE E R R E V I E W
The peer review history for this article is available at https://publo ns.com/publo n/10.1002/vms3.412.