Experimental Necrotic Enteritis Research Articles

Supplementation with organic yeast-derived selenium provides immune protection against experimental necrotic enteritis in broiler chickens

Necrotic enteritis (NE) is a potentially fatal poultry disease that causes enormous economic losses in the poultry industry worldwide. The study aimed to evaluate the effects of dietary organic yeast-derived selenium (Se) on immune protection against experimental necrotic enteritis (NE) in commercial broilers. Chickens were fed basal diets supplemented with different Se levels (0.25, 0.50, and 1.00 Se mg/kg). To induce NE, Clostridium perfringens (C. perfringens) was orally administered at 14 days of age post hatch. The results showed that birds fed 0.25 Se mg/kg exhibited significantly increased body weight gain compared with the non-supplemented/infected birds. There were no significant differences in gut lesions between the Se-supplemented groups and the non-supplemented group. The antibody levels against α-toxin and NetB toxin increased with the increase between 0.25 Se mg/kg and 0.50 Se mg/kg. In the jejunal scrapings and spleen, the Se-supplementation groups up-regulated the transcripts for pro-inflammatory cytokines IL-1β, IL-6, IL-8, iNOS, and LITAF and avian β-defensin 6, 8, and 13 (AvBD6, 8 and 13). In conclusion, supplementation with organic yeast-derived Se alleviates the negative consequences and provides beneficial protection against experimental NE.

Evaluation of the Immunoprotective Capacity of Five Vaccine Candidate Proteins against Avian Necrotic Enteritis and Impact on the Caecal Microbiota of Vaccinated Birds.

Avian necrotic enteritis is an enteric disease of broiler chickens caused by certain pathogenic strains of Clostridium perfringens in combination with predisposing factors. A vaccine offering complete protection against the disease has not yet been commercialized. In a previous study, we produced five recombinant proteins predicted to be surface-exposed and unique to necrotic enteritis-causing C. perfringens and the immunogenicity of these potential vaccine candidates was assessed in broiler chickens. In the current work, the relative contribution of the antibodies raised by these putative antigens to protect broiler chickens was evaluated using an experimental necrotic enteritis induction model. Additionally, the link between the immune response elicited and the gut microbiota profiles in immunized birds subjected to infection with virulent C. perfringens was studied. The ELISA results showed that the IgY antibody titers in vaccinated birds on days 21 and 33 were significantly higher than those on days 7 and 14 and those in birds receiving the adjuvant alone, while the relative contribution of the specific immunity attributed to these antibodies could not be precisely determined using this experimental necrotic enteritis induction model. In addition, 16S rRNA gene amplicon sequencing showed that immunization of birds with recombinant proteins had a low impact on the chicken caecal microbiota.

Effect of synbiotic supplementation on production performance and severity of necrotic enteritis in broilers during an experimental necrotic enteritis challenge

To evaluate the efficacy of synbiotic during a necrotic enteritis (NE) infection, a total of 360 day-old chicks were randomly assigned into 4 experimental groups in a 2×2 factorial setup: control, challenge, synbiotic (1 g/kg), and challenge+synbiotic, with 6 replicates. NE was induced by gavaging 1×104Eimeria maxima oocysts and 1×108 CFU/mL of Clostridium perfringens on d 14 (D14) and D19, 20, and 21, respectively. At D35, the NE challenge decreased the BW gain (P < 0.001) and increased feed conversion ratio (P=0.03), whereas synbiotic supplementation decreased the feed intake (P=0.04). At D21, NE challenge increased gut permeability (P < 0.001), decreased regulatory T cells (Tregs) in the cecal tonsil (CT) (P=0.02), increased Tregs in the spleen (P=0.02), decreased nitric oxide (NO) production in the spleen (P=0.04) and decreased IL-10 expression in CT (P=0.02), whereas synbiotic supplementation increased CD4+:CD8+ T cells in the spleen (P < 0.001) and decreased interferon (IFN)-γ expression in the jejunum (P=0.07), however, synbiotic supplementation during NE challenge decreased mid-gut lesion score (P < 0.001), increased CD4+:CD8+ T cells in CT and decreased IgA production in bile (P < 0.001), compared to the control group. At D28, synbiotic supplementation decreased CD4+:CD8+ T cells in CT (P < 0.001), whereas synbiotic supplementation during NE challenge decreased Tregs in CT (P < 0.001) and increased NO production in the spleen (P=0.04), compared to the control group. At D35, the NE challenge decreased CD4+:CD8+ T cells in the spleen (P=0.03), decreased IgA production in bile (P=0.02), decreased IL-10 expression in CT (P=0.04), and decreased IL-10 (P=0.009), IFN-γ (P=0.03) and inducible nitric oxide synthase (P=0.02) expression in the jejunum, whereas synbiotic supplementation increased Tregs in the spleen (P=0.04), compared to control group. Synbiotic supplementation during the NE challenge decreased both IL-1β (P=0.02) and IFN-γ (P=0.001) expression in CT, compared to the control group. It can be concluded that synbiotic supplementation increases production performance by decreasing mid-gut lesions and enhancing protective immunity against NE, and efficiency of synbiotic could be improved by blending additional probiotics and prebiotics.

Immunization of broiler chickens with recombinant alpha-toxin protein for protection against necrotic enteritis

Necrotic enteritis ( NE ), caused by Clostridium perfringens ( CP ), has been reported in a wide variety of avian species including chickens. Necrotic enteritis has been reported in most parts of the world with acute outbreaks occurring sporadically and most often in broiler chickens after maternal immunity declines with bird age. This study was conducted to develop computer-aided design of a recombinant CP protein vaccine in an experimental model of necrotic enteritis. A synthetic rCPA which has been designed using predictive tools expressed and obtained through cloning, used to immunize and assess intestinal lesion scores, antibody levels, growth performance, and CP reduction in vivo. Broilers were immunized on days 7, 14, and 21 (V groups) with the purified recombinant clostridium alpha-toxin. On days 24, 25, and 26, the birds were orally challenged with CP (1.0 × 10 9 CFU/bird). Increased body weight (BW), clearance of CP, increased antibodies, and intestinal lesion scores were measured as protection parameters after oral co-infection with CP. Birds immunized and challenged with recombinant protein (CHV) experienced significantly less body weight loss and less intestinal lesions compared to the positive control group (PC). Higher anti-recombinant anti-rCPA antibody titers were observed in the challenged vaccinated (CHV) and only vaccinated (V) groups with an increased level CP clearance of intestinal contents in immunized birds. Based on these results, vaccination with computer-aided clostridial recombinant proteins, particularly rCPA toxin, enhances protective immunity against experimental necrotic enteritis in broiler chickens.

The Effect of Necrotic Enteritis Challenge on Production Performance, Cecal Microbiome, and Cecal Tonsil Transcriptome in Broilers.

The objective of this study was to identify the effects of experimental necrotic enteritis (NE) infection on the production performance, gut microbiome, and cecal tonsil transcriptome in broiler birds. A total of 192 chicks were not-induced (control) or induced with NE. NE was induced by inoculating Eimeria maxima at 14 d of age and Clostridium perfringens at 19, 20, and 21 d of age. NE challenge increased (p < 0.01) NE lesion score at 7 days post-E.maxima infection (dpi), decreased (p < 0.01) average weight gain and increased (p < 0.01) mortality at 7 and 14 dpi. NE challenge increased (p < 0.05) gut permeability at 5, 6, and 7 dpi and increased ileal C. perfringens load at 5 dpi. NE challenge increased (p < 0.01) Eimeria oocyst shedding at 5, 6, 7, 8 and 14 dpi. NE challenge decreased (p < 0.05) the relative abundance of Lactobacillaceae and increased (p < 0.05) the relative abundance of Campylobacteriaceae, Comamonadaceae, and Ruminococcaceae at 6 dpi. NE challenge upregulated (p < 0.05) genes related to immune response and downregulated (p < 0.05) genes related to lipid metabolism at 6 dpi. It can be concluded that NE infection decreased beneficial bacteria and increased gut permeability.

Immunization with Pooled Antigens for Clostridium perfringens Conferred Partial Protection against Experimental Necrotic Enteritis in Broiler Chickens.

Necrotic enteritis (NE) is a multifactorial and important enteric infectious disease etiologically caused by pathogenic C. perfringens infection, accounting for the estimated loss of around USD 6 billion in the global poultry industry. The increasing incidence of NE was found to be associated with the voluntary reduction or withdrawal of antibiotic growth promoters from animal feed during recent years. Therefore, the development of effective vaccines specific to NE assumes a priority for the poultry industry. This study aimed to identify the potential C. perfringens proteins as vaccine targets for NE. Three recombinant C. perfringens proteins targeting five antigens were prepared: two chimeric proteins (alpha-toxin and NetB, fructose-1,6-bisphosphate aldolase (FBA) and a zinc metalloprotease (Zm)), and one single collagen adhesion protein (Cna). Their protection efficacies were evaluated with a potent challenge model of Eimeria maxima/C. perfringens dual infections using a netB+tpeL+ C. perfringens strain. Young chicks were immunized twice subcutaneously with adjuvanted C. perfringens proteins on Days 4 and 15. At six days after the second immunization, the chickens immunized with Cna, FBA, and Zm antigens, and alpha-toxin had much higher serum antibody titers than unvaccinated controls prior to the challenge. Following the challenge, the pooled antigen-immunized group demonstrated no mortality and the least lesion scores against virulent challenge. The results indicate that the immunization with multicomponent antigens, including C. perfringens housekeeping protein Cna, may confer partial protection.

Bacteriophage mediated control of necrotic enteritis caused by C. perfringens in broiler chickens.

In Egypt, little attention has been paid to the isolation and application of C. perfringens phages for treating necrotic enteritis at the farm level. This study aims to evaluate the efficiency of the podovirus C. perfringens phage in treating necrotic enteritis in broiler chickens. Accordingly, C. perfringens phage was isolated from cecal samples of apparently healthy chickens and characterized by transmission electron microscopy, thermal stability test, and pH stability test. Commercial 14-day-old Arbor Acres broiler chickens were allocated to three groups: group Ӏ received BHI broth and assigned as a negative control, group П served as a positive control group that was challenged with C. perfringens via oral gavage for four successive days, and group Ш was administrated six phage doses on several occasions after oral gavage challenge with C. perfringens. Daily clinical symptoms and mortality were recorded. At three-time intervals, necrotic enteritis lesions were scored. Cecal samples were examined for re-isolation and counting of C. perfringens. The isolated C. perfringens phage was a podovirus with an icosahedral head diameter of 78.7nm and a short non-contractile tail length of 22.2nm. It remained stable for 60min at 30°C and 50°C at pH values of 2, 4, 8, and 10. The phage-treated group (Ш) showed mild gross lesions with a lower mortality rate and reduced colony-forming units than the positive control group (П). The findings revealed that the isolated C. perfringens phage effectively treated experimental necrotic enteritis in broiler chickens.

Immunization with a subunit of housekeeping proteins provide partial protection against experimental necrotic enteritis in broiler chickens

Abstract Necrotic enteritis (NE) due to Clostridium perfringens (CP) infections is a prevalent enteric infectious disease accounting for $6 billion economical loss in global poultry industry. The increasing incidence of NE has been associated with the reduction and withdrawal of antibiotic growth promoters from animal feed during recent years. Therefore, the development of effective vaccines specific for NE assumes a priority for poultry industry. By far, no any effective vaccines are commercially available in broiler chickens. Our aim was to identify the potential CP proteins as vaccine targets for NE. We expressed 4 different recombinant CP proteins targeting 6 antigens: 3 major virulence-related toxins (alpha-toxin, NetB and TpeL), and 3 pilus subunit or enzymes (Collagen adhesion protein (Cna), Fructose-1,6-bisphosphate aldolase (FBA) and a Hypothetic protein (HP)), and their protection efficacies were evaluated with a severe Coccidiosis/NE challenge model using netB+tpeL+ CP strain. Young chicks were immunized twice subcutaneously with adjuvanted CP proteins on days 4, and 15. Prior to challenge, the chickens immunized with related antigens had much higher serum antibody titers for Cna and FBA/HP antigens than those with any of 3 toxin antigen-immunized groups. Following the challenge, the Cna-immunized group showed the least mortality and lowest lesion score among the individual antigen-vaccinated groups, while the pooled antigens-immunized group demonstrated no mortality against virulent challenges. The results indicate that the immunization with CP housekeeping protein, Cna, conferred better protection than other conventional virulence-related toxin proteins. Mucosal delivery system is being investigated next.

Developing an experimental necrotic enteritis model in turkeys - the impact of Clostridium perfringens, Eimeria meleagrimitis and host age on frequency of severe intestinal lesions

BackgroundNecrotic enteritis is a significant problem to the poultry industry globally and, in Norway up to 30% of Norwegian turkey grow-outs can be affected. However, despite an awareness that differences exist between necrotic enteritis in chickens and turkeys, little information exists concerning the pathogenesis, immunity, microbiota or experimental reproduction of necrotic enteritis in turkeys. In particular, it is important to determine the appearance of the gross lesions, the age dependency of the disease and the role of netB toxin of Clostridium perfringens. To this end, we report our findings in developing an in vivo experimental model of necrotic enteritis in turkeys.ResultsA four tier (0–3) scoring system with clearly defined degrees of severity of macroscopic intestinal lesions was developed, based on 2312 photographic images of opened intestines from 810 B.U.T. 10 or B.U.T. Premium turkeys examined in nine experiments. Loss of macroscopically recognizable villi in the anterior small intestine was established as the defining lesion qualifying for a score 3 (severe intestinal lesions). The developed scoring system was used to identify important factors in promoting high frequencies of turkeys with severe lesions: a combined Eimeria meleagrimitis and Clostridium perfringens challenge, challenge at five rather than 3 weeks of age, the use of an Eimeria meleagrimitis dose level of at least 5000 oocysts per bird and finally, examination of the intestines of 5-week-old turkeys at 125 to 145 h after Eimeria meleagrimitis inoculation. Numbers of oocysts excreted were not influenced by Clostridium perfringens inoculation or turkey age. Among three different lesion score outcomes tested, frequency of severe lesions proved superior in discriminating between impact of four combinations of Clostridium perfringens inoculation and turkey age at challenge.ConclusionsThis study provides details for the successful establishment of an in vivo model of necrotic enteritis in turkeys.

Elevated faecal ovotransferrin concentrations are indicative for intestinal barrier failure in broiler chickens

Intestinal health is critically important for the welfare and performance of poultry. Enteric diseases that cause gut barrier failure result in high economic losses. Up till now there is no reliable faecal marker to measure gut barrier failure under field conditions. Therefore, the aim of the present study was to identify a faecal protein marker for diminished intestinal barrier function due to enteric diseases in broilers. To assess this, experimental necrotic enteritis and coccidiosis in broilers were used as models for gut barrier failure. Ovotransferrin was identified as a marker for gut barrier failure using a proteomics approach on samples from chickens with necrotic enteritis. These results were confirmed via ELISA on samples derived from both necrotic enteritis and coccidiosis trials, where faecal ovotransferrin levels were significantly correlated with the severity of gut barrier failure caused by either coccidiosis or necrotic enteritis. This indicates that faecal ovotransferrin quantification may represent a valuable tool to measure gut barrier failure caused by enteric pathogens.

Effects of yeast cell wall on growth performance, immune responses and intestinal short chain fatty acid concentrations of broilers in an experimental necrotic enteritis model.

Subclinical necrotic enteritis (NE) causes devastating economic losses in the broiler chicken industry, especially in birds raised free of in-feed antibiotics. Prebiotics are potential alternatives to in-feed antibiotics. Yeast cell wall extract (YCW) derived from Saccharomyces cerevisiae is a prebiotic with known immune modulating effects. This study examined the effects of YCW and antibiotics (AB) during subclinical NE on broiler growth performance, intestinal lesions, humoral immune response and gut microflora metabolites. The study employed a 2 × 3 factorial arrangement of treatments. Factors were: NE challenge (yes or no) and feed additive (control, AB, or YCW). Each treatment was replicated in 8 floor pens with 15 birds per pen. Challenged birds had higher feed conversion ratio (FCR) than unchallenged birds on d 35 (P < 0.05). Dietary inclusion of AB decreased FCR regardless of challenge (P < 0.05) on d 24 and 35. Inclusion of YCW reduced serum interleukin-1 (IL-1) concentration in NE challenged birds (P < 0.01) and increased immunoglobulin (Ig) G (P < 0.05) and Ig M (P < 0.05) levels compared to other dietary treatments regardless of challenge. Yeast cell wall extract increased formic acid concentration in cecal contents during challenge and increased butyric acid concentration in unchallenged birds on d 16. This study indicates YCW suppressed inflammatory response, promoted generation of immunoglobulin and increased short chain fatty acid production suggesting potential benefits to bird health.

Variable protection against experimental broiler necrotic enteritis after immunization with the C-terminal fragment of Clostridium perfringens alpha-toxin and a non-toxic NetB variant

ABSTRACTNecrotic enteritis toxin B (NetB) is a pore-forming toxin produced by Clostridium perfringens and has been shown to play a key role in avian necrotic enteritis, a disease causing significant costs to the poultry production industry worldwide. The aim of this work was to determine whether immunization with a non-toxic variant of NetB (NetB W262A) and the C-terminal fragment of C. perfringens alpha-toxin (CPA247–370) would provide protection against experimental necrotic enteritis. Immunized birds with either antigen or a combination of antigens developed serum antibody levels against NetB and CPA. When CPA247–370 and NetB W262A were used in combination as immunogens, an increased protection was observed after oral challenge by individual dosing, but not after in-feed-challenge.

Effects of dietary selenium on host response to necrotic enteritis in young broilers

The effects of dietary supplementation of young broiler chickens with an organic selenium (Se) formulation, B-Traxim Se, on experimental necrotic enteritis (NE) were studied. Chickens treated with three Se doses (0.25, 0.50, 1.00 mg/kg) from hatch were orally challenged with Eimeria maxima at 14 days of age followed by Clostridium perfringens to induce NE. Chickens fed with 0.50 mg/kg Se showed significantly increased body weights and antibody levels against NetB, and significantly reduced gut lesions compared with non-supplemented chickens. However, there were no significant differences in Eimeria oocyst shedding between the Se-treated and non-supplemented groups. Levels of IL-1β, IL-6, IL-8, iNOS, LITAF, TNFSF15, AvBD6, AvBD8, and AvBD13 transcripts were increased in the gut and spleen of at least one of the three Se-treated groups compared with the non-treated group. These results suggest that dietary supplementation of young broilers with Se might be beneficial to reduce the negative consequence of NE.

Effects of in ovo injection with selenium on immune and antioxidant responses during experimental necrotic enteritis in broiler chickens

This study was conducted to investigate the effects of in ovo injection of Se on modulating the immune system and antioxidant responses in broiler chickens with experimental necrotic enteritis. Broiler eggs were injected at 18 d of embryo age with either 100 μL of PBS alone or sodium selenite (Na2 SeO3) in PBS, providing 0 (SS0), 10 (SS10), or 20 (SS20) μg of Se/egg. At 14 d posthatch, PBS-treated and uninfected chickens were kept as the control group, whereas the remaining chickens were orally infected with 1.0 × 104 sporulated oocysts of Eimeria maxima (SS0, SS10, SS20). At 18 d posthatch, E. maxima-infected chickens were orally infected with 1.0 × 109 cfu of Clostridium perfringens. Infected control SS0 group showed significantly decreased BW compared with the uninfected control. However, SS20 group showed significantly increased BW compared with the infected control SS0 group, whereas the BW were similar among uninfected control and infected SS10 and SS20 groups. The SS10 group showed significantly lower intestinal lesions compared with the SS0 group, and oocyst production was decreased in both SS10 and SS20 groups. Serum malondialdehyde level and catalase activity were also decreased in both SS10 and SS20 groups, whereas the superoxide dismutase level was significantly lower in the SS10 group compared with the SS0 group. The SS20 group showed significantly higher levels of transcripts for IL-1β and IL-6 in intestine, and SS10 and SS20 groups had higher levels of transcripts for IL-8 and inducible nitric oxide synthase expression and decreased glutathione peroxidase 7 mRNA levels compared with the SS0 group. The SS10 and SS20 groups also showed increased serum antibody levels to C. perfringens α-toxin and NetB toxin compared with the SS0 group. These collective results suggest that the injection of Se into the amniotic cavity of developing eggs may be beneficial for enhancing immune and antioxidant responses in the hatched chickens exposed to the necrotic enteritis-causing pathogens.

Temporary feed restriction partially protects broilers from necrotic enteritis

The objective of this study was to investigate the effect of feed restriction on the intestinal ecosystem and on the pathogenesis of experimental necrotic enteritis in broiler chicks. To induce subclinical necrotic enteritis, an experimental challenge model using a specific diet formulation, Gumboro vaccination, oral inoculation of broilers with a 10-fold dose of attenuated anticoccidial vaccine and multiple oral inoculations with a specific strain of Clostridium perfringens was adopted. Two hundred and forty 1-day-old Cobb 500 broilers were randomly allocated to four groups: feed restricted, challenged, both feed restricted and challenged, and negative control. At 21, 22, 23 and 24 days of age, the intestines, gizzard and liver were collected from 15 birds in each group and scored for gross lesions. The intestinal digesta was collected for pH and viscosity determination. One caecum from each bird was taken for microbiological analysis. The application of feed restriction in birds challenged with C. perfringens reduced the necrotic enteritis lesion score significantly (P ≤ 0.05) and feed restriction significantly reduced (P ≤ 0.05) pH in the small intestine, the viscosity of the jejunum digesta as well as the C. perfringens counts in the caeca compared with the controls. In conclusion, feed restriction of broilers has a positive effect on the intestinal ecosystem and a significant protective effect against necrotic enteritis in the subclinical experimental model.

The role of an attenuated anticoccidial vaccine on the intestinal ecosystem and on the pathogenesis of experimental necrotic enteritis in broiler chickens

The objective of the present study was to investigate the effect of an attenuated anticoccidial vaccination on the intestinal ecosystem and on the pathogenesis of experimental necrotic enteritis (NE) in broiler chickens. Two hundred and forty 1-day-old Cobb 500 broiler chickens were randomly allocated to four treatment groups according to the following experimental design: control Group N; Group PN, where birds were vaccinated with anticoccidial vaccine; Group M, where birds were challenged with Clostridium perfringens and with Eimeria maxima; and Group PM, where birds were both vaccinated and challenged. From each bird, the intestine, gizzard and liver were scored for gross NE lesions. Intestinal digesta were collected for pH and viscosity determination. Samples from the gastrointestinal tract and liver were taken for microbiological analysis. Evaluation of the experimental data revealed that Group M had significantly higher overall mean NE intestinal lesions compared with Group PM. Viscosity values of jejunum digesta as well as pH values of the duodenum, jejunum and ileum digesta in Group M were significantly lower compared with Group PM. C. perfringens counts in the caeca of Group PM were significantly lower compared with Group M. The milder decrease of pH and viscosity values of intestinal content and the reduction of C. perfringens counts in the caeca in challenged and vaccinated birds may explain the lower score of NE gross intestinal lesions and may suggest a positive effect on intestinal ecosystem and a significant protective effect of attenuated anticoccidial vaccination against NE in a subclinical experimental model.

Vaccination with Clostridium perfringens recombinant proteins in combination with Montanide™ ISA 71 VG adjuvant increases protection against experimental necrotic enteritis in commercial broiler chickens

This study was performed to compare four Clostridium perfringens recombinant proteins as vaccine candidates using the Montanide™ ISA 71 VG adjuvant in an experimental model of necrotic enteritis. Broiler chickens were immunized subcutaneously with purified clostridial recombinant NetB toxin, pyruvate: ferredoxin oxidoreductase (PFO), α-toxin, or elongation factor-Tu (EF-Tu), or with vehicle control, in conjunction with ISA 71 VG, and intestinal lesion scores, body weight gains, NetB toxin and PFO antibody levels, and proinflammatory cytokine and chemokine levels were measured as outcomes of protection following oral co-infection with C. perfringens and Eimeria maxima. Birds immunized with all recombinant proteins plus ISA 71 VG showed significantly reduced gut lesions compared with the ISA 71 VG-only group. Birds immunized with NetB toxin or PFO plus ISA 71 VG exhibited significantly increased body weight gains compared with the ISA 71 VG alone group. Greater NetB toxin antibody titers were observed in the NetB/ISA 71 VG group, and greater PFO antibody titers were evident in the PFO/ISA 71 VG group, each compared with the other three vaccine/adjuvant groups. Finally, decreased levels of gene transcripts encoding interleukin-8, tumor necrosis factor superfamily 15, and LPS-induced TNF-α factor were observed in the intestinal lymphocytes of chickens immunized with NetB toxin, PFO, α-toxin, and/or EF-Tu in the presence of ISA 71 VG compared with ISA 71 VG alone. All parameters evaluated were equal in co-infected chickens given ISA 71 VG alone compared with infected/adjuvant-free birds, indicating that the adjuvant itself did not have a disease protective effect. These results suggest that vaccination with clostridial recombinant proteins, particularly NetB toxin or PFO, in combination with ISA 71 VG enhances protective immunity against experimental necrotic enteritis in broiler chickens.

Control of Clostridium perfringens-induced necrotic enteritis in broilers by target-released butyric acid, fatty acids and essential oils

The efficacy of target-released butyric acid, medium-chain fatty acids (C6 to C12 but mainly lauric acid) and essential oils (thymol, cinnamaldehyde, essential oil of eucalyptus) micro-encapsulated in a poly-sugar matrix to control necrotic enteritis was investigated. The minimal inhibitory concentrations of the different additives were determined in vitro, showing that lauric acid, thymol, and cinnamaldehyde are very effective in inhibiting the growth of Clostridium perfringens. The in vivo effects were studied in two trials in an experimental necrotic enteritis model in broiler chickens. In the first trial, four groups of chickens were fed a diet supplemented with butyric acid, with essential oils, with butyric acid in combination with medium-chain fatty acids, or with butyric acid in combination with medium-chain fatty acids and essential oils. In all groups except for the group receiving only butyric acid, a significant decrease in the number of birds with necrotic lesions was found compared with the infected, untreated control group. In the second trial the same products were tested but at a higher concentration. An additional group was fed a diet supplemented with only medium-chain fatty acids. In all groups except for that receiving butyric acid in combination with medium-chain fatty acids and essential oils, a significant decrease in the number of birds with necrotic lesions was found compared with the infected, untreated control group. These results suggest that butyric acid, medium-chain fatty acids and/or essential oils may contribute to the prevention of necrotic enteritis in broilers.