Oregano (Origanum vulgare L.) Essential Oil Feed Supplement Protected Broilers Chickens against Clostridium perfringens Induced Necrotic Enteritis

Oregano (Origanum vulgare L.) is a well-known traditional medicine and a cooking spice. Recent practice has also applied the essential oil from oregano (OEO) in poultry due to its great potential for an antibiotic alternative. Our objective was to evaluate the potential effects of OEO (with carvacrol and thymol as the main active ingredient) on preventing necrotic enteritis (NE) caused by Clostridium perfringens (Cp) in chickens. In the feeding trial, a total of 450 one-day-old commercial Arbor Acres broilers were randomly assigned in 5 experimental groups during a 26-day production period (d19 to d 26 was the Cp challenge stage), and each group consisted of 6 replicate pens (15 birds each pen). All treatments were: basal diet (control group); basal diet and Cp challenge (model group); Cp challenge and 10 mg/kg enramycin (positive control group); Cp challenge and 200 mg/kg OEO product (OEO low dosage group, OEOL); Cp challenge and 300 mg/kg OEO product (OEO high dosage group, OEOH). OEO feed supplement at both dosages had significant effects on increasing the body weight gain (BWG) and reversing the dropped feed intake (FI) induced by Cp challenge. Histopathological changes in the ileums of broiler chickens with NE induced by Cp were alleviated by OEO, which was mutually confirmed by the intestinal lesion scores. Dosage did not influence the protective effect of OEO on intestinal lesion scores. Furthermore, OEO was found to have limited effects on tight junction-related gene expressions (Occludin and ZO-1). The broilers of the OEOL and OEOH groups significantly decreased the expression of TNF-α mRNA in the ileum and only the OEOH group was found to inhibit the IFN-γ expression of IFN- induced by Cp challenge. Finally, despite the fact that in vitro antibacterial effects by OEO were observed, considering its high minimum inhibitory concentration (MIC) value, we inferred that the protective effects by OEO against Cp challenge were not attributable to its direct antibacterial effects. We proposed OEO as a promising substitute for antibiotics against NE induced by Cp during poultry production.

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Supplemental Bacillus subtilis PB6 Improves Growth Performance and Gut Health in Broilers Challenged with Clostridium perfringens

Journal of Immunology Research ◽

10.1155/2021/2549541 ◽

2021 ◽

Vol 2021 ◽

pp. 1-11

Author(s):

Yan Liu ◽

Song Zhang ◽

Zheng Luo ◽

Dan Liu

Keyword(s):

Bacillus Subtilis ◽

Growth Performance ◽

Clostridium Perfringens ◽

Basal Diet ◽

Economic Losses ◽

Control Group ◽

Necrotic Enteritis ◽

Dosage Group ◽

Microbial Composition ◽

High Dosage Group

Clostridium perfringens (CP) is the principal pathogenic bacterium of chicken necrotic enteritis (NE), which causes substantial economic losses in poultry worldwide. Although probiotics are known to provide multiple benefits, little is known about the potential effects of Bacillus subtilis (B. subtilis) application in preventing CP-induced necrotic enteritis. In this study, 450 male Arbor Acres broilers were divided into 5 experimental treatments: A: basal diet (control group); B: basal diet and CP challenge (model group); C: CP challenge+10 mg/kg enramycin (positive control group); D: CP challenge+ 4 × 10 7 CFU / kg of feed B. subtilis PB6 (PB6 low-dosage group); and E: CP challenge+ 6 × 10 7 CFU / kg of feed B. subtilis PB6 (PB6 high-dosage group). There were 6 replicate pens per treatment with 15 broilers per pen. The present research examined the effect of Bacillus subtilis PB6 (B. subtilis PB6) on growth performance, mRNA expression of intestinal cytokines and tight junctions, and gut flora composition in broilers challenged with CP. The entire experiment was divided into two phases: the non-CP challenge phase (d0–18) and the CP challenge phase (d18–26). PB6 did not increase the growth performance during the first stage, but the PB6 high-dosage group was found to have larger body weight gain and ADFI during the CP challenge stage. Feed supplementation with PB6 reduced the lesion score of challenged chicks, with increased tight junction-related gene expression (occludin and ZO-1) and decreased TNF-α expression compared with CP-infected birds. A decrease in the abundance of Clostridium XI, Streptococcus, and Staphylococcus was observed after CP infection ( P < 0.05 ), while supplementation with PB6 restored the ileal microbial composition. In conclusion, administration of B. subtilis PB6 improved growth performance, enhanced intestinal barrier function, and mitigated intestinal inflammation/lesions, which might be due to its restoring effects on the ileal microbial composition in CP-challenged broilers.

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Probiotics Bacillus licheniformis Improves Intestinal Health of Subclinical Necrotic Enteritis-Challenged Broilers

Frontiers in Microbiology ◽

10.3389/fmicb.2021.623739 ◽

2021 ◽

Vol 12 ◽

Author(s):

Liugang Kan ◽

Fangshen Guo ◽

Yan Liu ◽

Van Hieu Pham ◽

Yuming Guo ◽

...

Keyword(s):

Bacillus Licheniformis ◽

Basal Diet ◽

Control Group ◽

Intestinal Damage ◽

Necrotic Enteritis ◽

Mrna Levels ◽

Poultry Production ◽

Intestinal Health ◽

Intestinal Lesion ◽

Mucin 2

Necrotic enteritis infection poses a serious threat to poultry production, and there is an urgent need for searching effective antibiotic alternatives to control it with the global ban on in-feed antibiotics. This study was conducted to investigate the effects of dietary Bacillus licheniformis replacing enramycin on the growth performance and intestinal health of subclinical necrotic enteritis (SNE)-challenged broilers. In total, 504 1-day-old Arbor Acres male chickens were selected and subsequently assigned into three treatments, including PC (basal diet + SNE challenge), PA (basal diet extra 10 mg/kg enramycin + SNE challenge), and PG (basal diet extra 3.20 × 109 and 1.60 × 109 CFU B. licheniformis per kg diet during 1–21 days and 22–42 days, respectively + SNE challenge). Results showed that B. licheniformis significantly decreased the intestinal lesion scores and down-regulated the Claudin-3 mRNA levels in jejunum of SNE-infected broilers on day 25, but increased the mucin-2 gene expression in broilers on day 42. In addition, B. licheniformis significantly up-regulated the mRNA levels of TRIF and NF-κB of SNE-challenged broilers compared with the control group on day 25 and TLR-4, TRIF compared with the control and the antibiotic group on day 42. The mRNA expression of growth factors (GLP-2 and TGF-β2) and HSPs (HSP60, HSP70, and HSP90) were up-regulated in B. licheniformis supplementary group on days 25 and 42 compared with group PC. LEfSe analysis showed that the relative abundance of Lachnospiraceae_UCG_010 was enriched in the PG group; nevertheless, Clostridiales_vadinBB60 and Rnminococcaceae_NK4A214 were in PA. PICRUSt analysis found that the metabolism of cofactors and vitamins, amino acid metabolism, and carbohydrate metabolism pathways were enriched, whereas energy metabolism, membrane transport, cell motility, and lipid metabolism were suppressed in B. licheniformis-supplemented groups as compared with the PC control. In conclusion, dietary supplementation of B. licheniformis alleviated the intestinal damage caused by SNE challenge that coincided with modulating intestinal microflora structure and barrier function as well as regulating intestinal mucosal immune responses.

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Supplementing Garlic Nanohydrogel Optimized Growth, Gastrointestinal Integrity and Economics and Ameliorated Necrotic Enteritis in Broiler Chickens Using a Clostridium perfringens Challenge Model

Animals ◽

10.3390/ani11072027 ◽

2021 ◽

Vol 11 (7) ◽

pp. 2027

Author(s):

Doaa Ibrahim ◽

Tamer Ahmed Ismail ◽

Eman Khalifa ◽

Shaimaa A. Abd El-Kader ◽

Dalia Ibrahim Mohamed ◽

...

Keyword(s):

Growth Performance ◽

Clostridium Perfringens ◽

Broiler Chickens ◽

Body Weight Gain ◽

Economic Benefits ◽

Production Costs ◽

Broiler Chicks ◽

Necrotic Enteritis ◽

Feed Conversion ◽

Intestinal Lesion

Necrotic enteritis (NE) caused by Clostridium perfringens (C. perfringens) results in impaired bird growth performance and increased production costs. Nanotechnology application in the poultry industry to control NE outbreaks is still not completely clarified. Therefore, the efficacy of dietary garlic nano-hydrogel (G-NHG) on broilers growth performance, intestinal integrity, economic returns and its potency to alleviate C. perfringens levels using NE challenge model were addressed. A total of 1200 male broiler chicks (Ross 308) were assigned into six groups; four supplemented with 100, 200, 300 or 400 mg of G-NHG/kg diet and co-challenged with C. perfringens at 21, 22 and 23 d of age and two control groups fed basal diet with or without C. perfringens challenge. Over the total growing period, the 400 mg/kg G-NHG group had the most improved body weight gain and feed conversion efficiency regardless of challenge. Parallel with these results, the mRNA expression of genes encoding digestive enzymes (alpha 2A amylase (AMY2A), pancreatic lipase (PNLIP) and cholecystokinin (CCK)) and intestinal barriers (junctional adhesion molecule-2 (JAM-2), occludin and mucin-2 (Muc-2)) were increased in groups fed G-NHG at higher levels to be nearly similar to those in the unchallenged group. At 14 d post challenge, real-time PCR results revealed that inclusion of G-NHG led to a dose-dependently decrease in the C. perfringens population, thereby decreasing the birds’ intestinal lesion score and mortality rates. Using 400 mg/kg of G-NHG remarkably ameliorated the adverse effects of NE caused by C. perfringens challenge, which contributed to better growth performance of challenged birds with rational economic benefits.

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Evaluation of the effects of Salvia officinalis essential oil on plasma biochemistry, gut mucus and quantity of acidic and neutral mucins in the chicken gut

Acta veterinaria ◽

10.2478/acve-2014-0014 ◽

2014 ◽

Vol 64 (1) ◽

pp. 138-148 ◽

Cited By ~ 2

Author(s):

Adriana Čapkovičová ◽

Zuzana Maková ◽

Elena Piešová ◽

Anabela Alves ◽

Štefan Faix ◽

...

Keyword(s):

Essential Oil ◽

Plasma Cholesterol ◽

Basal Diet ◽

Goblet Cells ◽

Salvia Officinalis ◽

Control Group ◽

Mucus Layer ◽

Plasma Biochemistry ◽

Internal Organs ◽

Salvia Officinalis L

Abstract In this study the effects of Salvia officinalis L. essential oil on the thickness of the gut mucus layer and quantity of neutral and acidic mucins in chickens were observed. Oneday-old chickens of Isa Brown breed were divided into five groups (n = 9) and fed 11 weeks as follows: control group (C): basal diet; experimental groups (E1 - E4) - same as C + sage essential oil (EO) in concentrations of 0.01%; 0.025%; 0.05% and 0.1%, respectively. The thickness of the mucus layer in the duodenum significantly increased in E3(0.05% sage EO) compared to E1 (0.01 % sage EO). In E3 (0.05% sage EO) the number of goblet cells containing acidic and neutral mucins was significantly decreased in the duodenum and jejunum and increased in the ileum compared to C. Feeding the diet supplemented with Se and 0.01% sage EO (E1) decreased plasma cholesterol level in comparison with E3(0.05 % sage EO). The addition of 0.05% (E3) and 0.1% sage EO (E4) to the diet caused a decrease in calcium plasma level compared to E2 (0.025 % sage EO). Plasma glucose level was significantly decreased in groups fed 0.05% (E3) and 0.025% sage EO (E2) compared with 0.01% sage (E1). The weight of internal organs was not affected by the diets. Our results suggest that effects of sage EO on the adherent mucus layer dynamics and mucin type distribution in the chicken intestine are dependent on sage EO dose and intestinal segment and there is still a need for further studies in order to obtain a plausible explanation.

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Protective Effects of Lactobacillus plantarum 16 and Paenibacillus polymyxa 10 Against Clostridium perfringens Infection in Broilers

Frontiers in Immunology ◽

10.3389/fimmu.2020.628374 ◽

2021 ◽

Vol 11 ◽

Author(s):

Li Gong ◽

Baikui Wang ◽

Yuanhao Zhou ◽

Li Tang ◽

Zihan Zeng ◽

...

Keyword(s):

Lactobacillus Plantarum ◽

Clostridium Perfringens ◽

Intestinal Microbiota ◽

Metabolic Pathways ◽

Paenibacillus Polymyxa ◽

Basal Diet ◽

Control Group ◽

No Production ◽

Jejunal Mucosa ◽

Protective Effects

This study aimed to investigate the protective effects of Lactobacillus plantarum 16 (Lac16) and Paenibacillus polymyxa 10 (BSC10) against Clostridium perfringens (Cp) infection in broilers. A total of 720 one-day-old chicks were randomly divided into four groups. The control and Cp group were only fed a basal diet, while the two treatment groups received basal diets supplemented with Lac16 (1 × 108 cfu·kg−1) and BSC10 (1 × 108 cfu·kg−1) for 21 days, respectively. On day 1 and days 14 to 20, birds except those in the control group were challenged with 1 × 108 cfu C. perfringens type A strain once a day. The results showed that both Lac16 and BSC10 could ameliorate intestinal structure damage caused by C. perfringens infection. C. perfringens infection induced apoptosis by increasing the expression of Bax and p53 and decreasing Bcl-2 expression and inflammation evidence by higher levels of IFN-γ, IL-6, IL-1β, iNOS, and IL-10 in the ileum mucosa, and NO production in jejunal mucosa, which was reversed by Lac16 and BSC10 treatment except for IL-1β (P < 0.05). Besides, the two probiotics restored the intestinal microbiota imbalance induced by C. perfringens infection, characterized by the reduced Firmicutes and Proteobacteria and the increased Bacteroidetes at the phyla level and decreased Bacteroides fragilis and Gallibacterium anatis at the genus level. The two probiotics also reversed metabolic pathways of the microbiota in C. perfringens-infected broilers, including B-vitamin biosynthesis, peptidoglycan biosynthesis, and pyruvate fermentation to acetate and lactate II pathway. In conclusion, Lac16 and BSC10 can effectively protect broilers against C. perfringens infection through improved composition and metabolic pathways of the intestinal microbiota, intestinal structure, inflammation, and anti-apoptosis.

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A Carvacrol-Rich Essential Oil Extracted From Oregano (Origanum vulgare “Hot & Spicy”) Exerts Potent Antibacterial Effects Against Staphylococcus aureus

Frontiers in Microbiology ◽

10.3389/fmicb.2021.741861 ◽

2021 ◽

Vol 12 ◽

Author(s):

Yuanpeng Hao ◽

Jingyi Li ◽

Lei Shi

Keyword(s):

Staphylococcus Aureus ◽

Essential Oil ◽

Cell Damage ◽

Mass Spectrometry Analysis ◽

Gas Chromatography Mass Spectrometry ◽

Major Constituent ◽

Lag Phase ◽

Pathway Enrichment Analysis ◽

Antibacterial Effects ◽

Origanum Vulgare

Oregano essential oil (OEO), as a natural antimicrobial, has gained increased interest from food researchers and manufacturers. However, a few studies have investigated its possible antibacterial effects against Staphylococcus aureus using the proteomic tool. The present study aimed to explore the antibacterial effect and mechanism of a carvacrol-rich OEO extracted from Origanum vulgare “Hot & Spicy” on the inactivation of S. aureus. The gas chromatography–mass spectrometry analysis of the OEO allowed the detection of 27 compounds; the major constituent was carvacrol (84.38% of total compounds). The average diameter of the inhibitory zone (DIZ) value was 29.10 mm, and the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of OEO against S. aureus were 0.125 and 0.25 mg/mL, respectively. The growth curve assay indicated that the OEO prolonged the lag phase of S. aureus. The decrease in cell viability, changes in the integrity of cell membrane, and abnormal cell morphology further reflected the cell damage of S. aureus caused by the OEO. In addition, a label-free proteomic analysis was applied to analyze the regulatory networks of S. aureus in response to 1/2 MIC OEO-treatment stress. Of the 56 differentially expressed proteins (DEPs) identified, 26 were significantly upregulated and 30 downregulated. The Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis indicated that the DEPs were mainly involved in pathways of ribosomes; valine, leucine, and isoleucine biosynthesis; and phenylalanine, tyrosine, and tryptophan biosynthesis, which suggested that the growth inhibition of S. aureus might be due to the disordered effect of the OEO on protein synthesis and amino acid metabolism. These findings deepened our understanding regarding S. aureus survival and metabolism responses to the OEO treatment and suggested that the carvacrol-rich OEO could be used in food production environments to effectively control S. aureus.

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The dietary supplement efficiency of essential oil of chive (Allium macrostemon) on the productivity and health performance of broilers

Can Tho University Journal of Science ◽

10.22144/ctu.jen.2020.018 ◽

2020 ◽

Vol 12 (3) ◽

Author(s):

Phan Vũ Hải ◽

Pham Hoang Son Hung ◽

Hò Thị Dung ◽

Tràn Ngọc Liem ◽

Nguyẽn Dinh Thùy Khuong ◽

...

Keyword(s):

Weight Gain ◽

Essential Oil ◽

Dietary Supplement ◽

Basal Diet ◽

Control Group ◽

Growth Promoter ◽

Broiler Production ◽

Health Performance ◽

Allium Macrostemon ◽

Diarrhea Syndrome

This experiment was conducted to study on the effect of chive essential oil (Allium macrostemon) (CEO) on the production and health of broilers. One-day old roosters (Ross 308) were allocated to 4 treatments with 3 replications (10 birds/replication). The experiment consisted of control group (CT) with a basal diet and T1, T2 and T3 groups with the supplement in the basal diet by levels of CEO at 100, 300 and 500 mg/kg respectively. In general, boilers supplemented with CEO had the better FCR compared to the CT. However, broilers in T2 group (300 mg/kg) showed the highest efficiency in terms of weight gain, FCR and PEI. In addition, the use of CEO in T2 and T3 group tended to reduce the incidence of respiratory and diarrhea syndrome in broilers by more than 50% of cases. It can be concluded that the supplement of CEO at the level of 300 mg/kg in the diet had improved the productive and health performance and may be a viable alternative to growth promoter in broiler production.

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Oregano Essential Oil (Origanum vulgare L.), as a Food-supplement in a Rearing of Piglets

Scientific Messenger of LNU of Veterinary Medicine and Biotechnology ◽

10.32718/nvlvet9510 ◽

2019 ◽

Vol 21 (95) ◽

pp. 55-61 ◽

Cited By ~ 1

Author(s):

I. Salamon ◽

J. Poracova ◽

M. Hrytsyna

Keyword(s):

Escherichia Coli ◽

Essential Oil ◽

Lactobacillus Plantarum ◽

Physiological Effect ◽

Feed Additives ◽

The Body ◽

Control Group ◽

Origanum Vulgare ◽

Opportunistic Bacteria ◽

Oil Extract

The article describes the results of the study of the physiological effect on the organism of pigs - mammals and weaned piglets. Feed additives were made from the essential oil of Origanum vulgare “Aditívum do kŕmnych zmesí na báze pamajoránu”. It has been established that O. vulgare dry grass contains essential oils (0.15–0.50%), where carvacrol, thymol, cymol and other terpenes are main components. Extract of O. vulgare essential oil has a pronounced inhibitory effect on the enteropathogenic bacteria Escherichia coli hemolytic and Salmonella enteritica var. Enteritidis. In quantifying, it was found that 20% of the essential oil extract has inhibition role, which is 4 times more effective than 100%, has the largest inhibitory role. The resistant symbiotic bacterium of the gut Enterococcus faecium M-7 was not affected by essential oil. The following indicators were used to establish the physiological effect of herbal supplements on the body of piglets: increasment in body weight, resistance of piglets to diseases, the state of intestinal microflora. A control and two experimental groups were added for the experiment, where the phyto-additive was included in to the feed: the first consisted of bacteria Lactobacillus plantarum with 0.05% extract of O. vulgare (LO) essential oil and the second – only 0.05% essential oil extract О. vulgare. (O). In the two experimental groups, the pigs' health was good and their mortality was not observed either before or after weaning. The incidence was 18% before weaning and 36% after weaning in the control group. The average daily weight gaining was significant in both groups (31–36%). Moreover, piglets were larger at birth and before weaning they increased in weight to 8 times, whereas over the next 24 days their weight increased only to 50%. The difference in gaining weight between the two expirimantal groups was insignificant. The use of the herbal supplement LO is more effective than O, because the stability index of the gut microbiota gradually increases from 2.2 to 3.37 due to decrease in the number of opportunistic bacteria Escherichia coli and enterobacteria and growth of lactobacilli. Therefore, a phyto-supplement consisting of Lactobacillus plantarum bacteria with 0.05% O. vulgare (LO) essential oil extract can be recommended as a probiotic to feed mammals and weaned pigs for improving health and disease prevention.

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Antimicrobial activity of thyme (Thymus vulgaris) and oregano (Origanum vulgare) essential oils against Listeria monocytogenes in fermented sausages

IOP Conference Series Earth and Environmental Science ◽

10.1088/1755-1315/854/1/012064 ◽

2021 ◽

Vol 854 (1) ◽

pp. 012064

Author(s):

Drago Nedic ◽

Nevena Grkovic ◽

Vesna Kalaba ◽

Bojan Golic ◽

Tanja Ilic ◽

...

Keyword(s):

Listeria Monocytogenes ◽

Essential Oil ◽

Essential Oils ◽

Detection Threshold ◽

Thymus Vulgaris ◽

Antibacterial Effects ◽

High Concentration ◽

Origanum Vulgare ◽

Fermented Sausages ◽

Microbiological Status

Abstract The aim of this study was to investigate antibacterial effects of oregano and thyme essential oils on Listeria monocytogenes in fermented sausages and their effect on the sensory characteristics of these sausages. For testing purposes, sausages contaminated with L. monocytogenes were produced. Changes in the microbiological status of fermented sausages and physicochemical properties were monitored during ripening. Essential oils exhibited antibacterial activity against L. monocytogenes, and in the groups with a high concentration (0.6%) of oregano or thyme essential oils (KLO2 and KLT2), the number of L. monocytogenes was below the detection threshold on day 14 of ripening, with a stronger effect of oregano. In groups with 0.3% essential oil of oregano or thyme added, the number of L. monocytogenes was reduced to below the detection threshold on day 21 of ripening. During the ripening, the aw and pH of all test groups of fermented sausages decreased. Experimental sausages with 0.3% thyme essential oil had acceptable smell and taste, while in other experimental groups, sausage smell and taste were very intense, uncharacteristic and unacceptable.

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Oral immunization of broilers with chitosan nano-encapsulated extracellular and cell wall proteins of necrotic enteritis-induced Clostridium perfringens

10.1101/2020.10.23.351817 ◽

2020 ◽

Author(s):

Nour Muinis Ramadan ◽

Gabriel Akerele ◽

Sankar Renu ◽

Gourapura Renukaradhya ◽

Ramesh Selvaraj

Keyword(s):

Cell Wall ◽

Clostridium Perfringens ◽

Neutralizing Antibodies ◽

Body Weight Gain ◽

Oral Immunization ◽

Extracellular Proteins ◽

Control Group ◽

Necrotic Enteritis ◽

Cell Wall Proteins ◽

Feed Conversion

AbstractCurrently, there is no commercial vaccine to control Clostridium perfringens (CP) or necrotic enteritis – a clinically and economically devastating disease. Two chitosan-nanoparticle encapsulated CP (CNP-CP) vaccines using extracellular proteins (ECP) and cell wall proteins (CWP) were synthesized: a CNP-CP ECP + CWP vaccine (SC vaccine) and a CNP-CP ECP + CWP surface-adsorbed CWP vaccine (SCC vaccine). The experiment comprised a complete randomized design of 3 treatments replicated 5 times: SC, SCC and nonimmunized control. Broilers orally gavaged with SC or SCC vaccine were primed and boosted with 70μg dose at 3- and 14-days post-hatch (dph). SDS-PAGE analysis revealed bands at 54.7 and 84.7 kDa in the ECP and 17 bands for CWP fraction. There were no differences in body weight gain, feed conversion ratio and mortality rate between treatments. At 17dph, serum of birds in the SC and SCC groups had higher neutralizing antibodies (P<0.01) compared to the control group. At 17dph, there was an increase in bile anti-ECP IgA levels in the SC vaccinated birds and a non-significant increase in SCC vaccinated birds compared to control. At 17dph, bile specific anti-CP CWP IgA levels were greater (P<0.05) in both immunized groups compared to control. At 17 and 21dph, serum anti-ECP and anti-CP CWP IgY levels in SC and SCC birds were comparable to the control. At 21dph, CD4+/CD8+ T-cell ratio in SC and SCC vaccinated groups were elevated (P ≤ 0.01) compared to control. At 17dph, SC and SCC vaccinated birds had a significant reduction (P ≤ 0.001) in α-toxin levels in caecal contents compared to control. Caecal α-toxin levels remained reduced at 21dph (P < 0.05) in SC birds and numerically reduced in SCC vaccinated birds compared to control. Jejunal CP load in SCC birds was significantly reduced by 1.4 Log10 copy numbers of CP/g compared to control and no differences were observed in liver CP load between immunized and non-immunized birds. SC and SCC immunization did not alter TNF-α, IL-10, iNOS, IL-6 or IL-17 mRNA expression. At 17 and 21dph, SC and SCC immunized birds demonstrated greater sera bactericidal activity compared to control. CNP-SC and CNP-SCC immunization induced specific immune response against C. perfringens and reduced CP colonization via oral route of administration.

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