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Sodium Butyrate in Broiler
Contents
 Host defense peptides HDPs.
 Host defense peptides HDP and sodium butyrate.
 Trophic effect of sodium butyrate.
 Antibacterial effect of sodium butyrate.
 Other effects of sodium butyrate.
Contents
 Host defense peptides HDPs
 Host defense peptides HDP and sodium butyrate
 Trophic effect of sodium butyrate
 Antibacterial effect of sodium butyrate
 Other effects of sodium butyrate
What is Host Defense Peptides HDPs?
Host defense peptides (HDPs), also known as antimicrobial
peptides, are present in all species and constitute a critical
component of the innate immunity.
Cont. …
 There are two major families of HDPs in vertebrates;
1. Defensins
2. Cathelicidins
 The chicken genome was recently found to encode a total of;
1) 14 b-defensins known as AvBD1-14
2) 4 cathelicidins, namely fowlicidins 1–3 and cathelicidin-
B1.
Cont. …
 In chicken, AvBDs and cathelicidins are expressed in a wide
range of tissues;
– Bdefensins in liver , digestive, respiratory, and reproductive
tracts.
– Cathelicidins in bone marrow or bursa.
Therapeutic Effects
Of Host Defense Peptides
1. Antimicrobial effect
2. Immune stimulatory effect
Host Defense Peptides
Antimicrobial Activities
HDPs possess broad-spectrum antimicrobial activities against:
1. Bacteria
2. Protozoa
3. Enveloped virus
4. Fungi
mainly through direct binding and lysis of microbial membranes.
Cont. …
 Because of such physical interactions, it is extremely difficult
for pathogens to develop resistance to HDPs.
 Many chicken HDPs have been found to possess potent
antibacterial activities against a broad range of bacteria
including Salmonella.
Host Defense Peptides
Immune Stimulation
 HDPs have a strong capacity to modulate the innate immune
response by inducing chemotaxis and activation of various
types of leukocytes.
Host Defense Peptides
 Because of these pleiotropic effects, HDPs have been actively
explored as a new class of therapeutic agents against
antibiotic-resistant microbes and other inflammatory
diseases.
Contents
 Host Defense Peptides HDPs
 Host Defense Peptides HDP and Sodium Butyrate
 Trophic Effect Of Sodium Butyrate
 Antibacterial Effect Of Sodium Butyrate
 Other effects Of Sodium Butyrate
In Vivo Induction Of
The Avbd9 Gene Expression
In The Intestinal Tract Of Chickens By Butyrate
Sodium Butyrate and HDPs
 Butyrate, a major species of short-chain fatty acids produced
by bacterial fermentation of undigested carbohydrates in the
intestine, was recently found to be capable of inducing HDP
expression in humans, rabbits and poultry.
Study Design
Birds profile:
Two-day-old male Cornish Rock broilers
Treatment
Feed with standard ration with or without supplementation of
0.1% and 0.2% butyrate for 2 days.
Evaluation:
Crop, cecal tonsil and cecum were collected from each bird and
the AvBD9 gene expression was evaluated by real-time PCR.
Reduction of the S. enteritidis titer in the cecal
contents of chickens following oral supplementation
of butyrate
Study Objective
 To examine the effect of supplementing butyrate in the feed
on the titer of Salmonella enteritidis in the cecum following
experimental infections.
Study Design
Birds profile:
20-day old male broilers were equally divided into two groups,
each of 10.
Treatment
Feed with a standard antibiotic-free diet mixed with and without
0.2% sodium butyrate for 2 days.
Cont. …
Challenge
Birds were then inoculated with 1X10 log 6 CFU of S. enteritidis
phage type 13a and continued with butyrate feeding for another
4 days.
Evaluation
The S. enteritidis titer in the cecal content was quantitated from
each bird by serial plating on Brilliant Green agar plates
containing 20 mg/ml nalidixic acid.
Study Design
 This trial was similarly conducted with an additional group of
10 broilers fed with 0.1% butyrate.
 Each dot represents the bacterial titer from a bird and the
solid line represents the median value of each treatment.
 Brackets indicate the statistical significance of differences
Contents
 Host Defense Peptides HDPs
 Host Defense Peptides HDP and Sodium Butyrate
 Trophic Effect Of Sodium Butyrate
 Antibacterial Effect Of Sodium Butyrate
 Other effects Of Sodium Butyrate
Trophic Effect Of Sodium Butyrate
 Sodium butyrate is a preferred source of energy for the
enterocytes.
 This will result in:
1. Better development of the intestinal villi.
2. Stronger gut lining.
Study Design
Trial Objective
To evaluate the influence of sodium butyrate on digestibility of a
corn-soya diet, as well as villi development.
Birds Profile
A total of 750 Cobb 400 one day old broiler chickens.
Treatments
Cont. …
The trial evaluated 3 treatments:
1. Control (no additives)
2. 0.5 kg/t of Product X (92% sodium butyrate)
3. 1 kg/t of Product X.
Evaluations
1. The digestible energy of the diet
2. The digestibility of the protein
3. The lengths and widths of the villis
Digestibility of Energy
Energy kcal/kg
3,105
3,264
3,285
Digestibility of Energy
Control Product X 0.5g/kg Product X 1.0g/kg
Protein Digestibility
Protein Digestability
65%
66%
68%
Protein Digestibility
Control Product X 0.5g/kg Product X 1.0g/kg
Villi Length
21 Days Old mm
Villi Length 21 Days Old mm
0.98
1.52
1.24
Villi Length 21 Days Old mm
Control Product X 0.5g/kg Gustor 1.0g/kg
Villi Length
42 Days Old mm
Villi Length 42 Days Old mm
0.94
1.31
1.12
Villi Length 42 Days Old mm
Control Product X 0.5g/kg Gustor 1.0g/kg
Villi Width
21 Days Old µm
Villi Width 21 Days Old µm
224.70
260.80
197.70
Villi Width 21 Days Old µm
Control Product X 0.5g/kg Gustor 1.0g/kg
Villi Width
42 Days Old µm
Villi Width 21 Days Old µm
138.50
314.20 317.20
Villi Width 42 Days Old µm
Control Product X 0.5g/kg Gustor 1.0g/kg
Conclusion
 The digestible energy of the diet and the digestibility of the
protein were significantly improved by the addition of sodium
butyrate.
 The lengths and widths of the villi were also positively
affected by the addition of butyrate in the diet.
 From this trial and previous research, it was concluded that
inclusion of butyrate in the diet improves the digestibility of
energy and protein by increasing intestinal absorption surface
Contents
 Host defense peptides HDPs
 Host defense peptides HDP and sodium butyrate
 Trophic effect of sodium butyrate
 Antibacterial effect of sodium butyrate
 Other effects of sodium butyrate
VFA
Antibacterial Effect
 It has been demonstrated that VFAs can inhibit the growth of
bacteria of the group of Enterobacteriaceae (Salmonella,
Escherichia coli …).
Mechanism of action
 This is because the undissociated form of these acids can
freely diffuse across the bacterial membrane.
 Once inside the cytoplasm of the bacteria, the acid
dissociates, thus releasing free hydrogen ion and reducing the
pH, which causes internal cell damage.
Butyric Acid
 In this respect, butyric acid performs very well.
 Hume et al. (1993) showed that butyric acid has a higher
diffusion coefficient than other acids with a shorter chain,
which allows it to pass through the bacterial membrane more
easily.
 Galfi and Neogradi (1995) found that the concentration of
butyrate required to reduce the growth of E. coli by 50% is
much lower than the concentration of the other volatile fatty
acids, acetate and propionate.
Butyric Acid Vs Other Fatty Acids
Van Immerseel (2004) observed that
butyric acid was the most effective
acid to help controlling Salmonella
enteritidis in broilers after a challenge.
Number of colony forming units (CFU) per gram of ceca
at day 8 after hatch of chickens orally inoculated with
1,000 CFU Salmonella enteritidis at days 5 and 6 and
given feed supplemented with different organic acids
(Adapted from Van Immersel, 2004)
Butyric Acid
Selective Action
 It should be also noted that the antibacterial effect of
butyrate is selective.
 For instance, in 1990, Galfi and Bokori demonstrated that
butyric acid favors the development of lactic flora which
competes against the enteropathogenic one (Clostridia, E.
coli, Coccidiae…).
Butyric Acid
Coccidia
 Leeson et al. (2005) also observed that birds receiving butyric
acid before coccidial oocyte challenge showed higher weight
gain following the challenge compared with birds that
received a control feed.
Butyric Acid
Salmonella
 One very unique feature of butyric acid is its ability to
negatively affect the virulence of pathogenic bacteria such as
Salmonella.
Cont. …
 To infect an animal host, Salmonella must penetrate the
intestinal epithelial barrier.
 This very important step is called invasive phase and requires
specific genes inside the bacterium.
 The invasion genes are located on the so-called
“pathogenicity island” and are encoding some regulatory
proteins, some structural components of a needle complex,
plus some effector proteins, which make possible the entry of
Salmonella into the epithelial cells.
Cont. …
 The pathogenicity island itself is activated by another protein
called HilA, which can be environmentally regulated.
 Different publications have shown that butyrate and
propionate can reduce the pathogenicity of Salmonella
Typhimurium or Salmonella Enteritidis, whereas other acids
such as acetic or formic are actually promoting the invasion of
epithelium cells by the bacteria (Durant et al., 2000 - Van
Immerseel et al., 2003 - Lawhon et al., 2005 - Gantois et al.,
2006 - Huang et al., 2008).
Cont. …
 The exact mode of action of butyrate in the bacterial cell is
still unknown but it seems it can interfere with the genetic
expression and results in a down regulation of the
pathogenicity island, regulating the invasive phenotypes of
Salmonella.
Contents
 Host defense peptides HDPs
 Host defense peptides HDP and sodium butyrate
 Trophic effect of sodium butyrate
 Antibacterial effect of sodium butyrate
 Other effects of sodium butyrate
Other effects Of Sodium Butyrate
1. Stimulation of Pancreatic Secretions
 Although other organic acids also produced this effect,
butyrate seems to be the one that causes the greatest
stimulation of pancreatic secretions.
 Katoh and Tsudo (1984) observed that the injection of
sodium butyrate in pigs resulted in greater stimulation of
pancreatic fluid secretion and increased amylase
secretion.
Cont. …
 Katoh et al. (1989) demonstrated that intravenous injection of
sodium butyrate in calves resulted in a higher secretion of
pancreatic juice and greater release of proteins (particularly
amylase) when compared to acetate and propionate.
Cont. …
2. Stimulation of Insulin Secretions
 Sano et al. (1995) also observed that the administration of
butyrate caused an increase in the plasmatic levels of
insulin, which normally results in higher amount of energy
and proteins deposited in the animal tissues.
Cont. …
3. Improves Absorption Of Electrolytes
 Sodium butyrate also improves the absorption of
electrolytes and reduces the incidence of diarrhoea (Galfi,
1989).

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Sodium butyrate in broiler

  • 2. Contents  Host defense peptides HDPs.  Host defense peptides HDP and sodium butyrate.  Trophic effect of sodium butyrate.  Antibacterial effect of sodium butyrate.  Other effects of sodium butyrate.
  • 3. Contents  Host defense peptides HDPs  Host defense peptides HDP and sodium butyrate  Trophic effect of sodium butyrate  Antibacterial effect of sodium butyrate  Other effects of sodium butyrate
  • 4. What is Host Defense Peptides HDPs? Host defense peptides (HDPs), also known as antimicrobial peptides, are present in all species and constitute a critical component of the innate immunity.
  • 5. Cont. …  There are two major families of HDPs in vertebrates; 1. Defensins 2. Cathelicidins  The chicken genome was recently found to encode a total of; 1) 14 b-defensins known as AvBD1-14 2) 4 cathelicidins, namely fowlicidins 1–3 and cathelicidin- B1.
  • 6. Cont. …  In chicken, AvBDs and cathelicidins are expressed in a wide range of tissues; – Bdefensins in liver , digestive, respiratory, and reproductive tracts. – Cathelicidins in bone marrow or bursa.
  • 7. Therapeutic Effects Of Host Defense Peptides 1. Antimicrobial effect 2. Immune stimulatory effect
  • 8. Host Defense Peptides Antimicrobial Activities HDPs possess broad-spectrum antimicrobial activities against: 1. Bacteria 2. Protozoa 3. Enveloped virus 4. Fungi mainly through direct binding and lysis of microbial membranes.
  • 9. Cont. …  Because of such physical interactions, it is extremely difficult for pathogens to develop resistance to HDPs.  Many chicken HDPs have been found to possess potent antibacterial activities against a broad range of bacteria including Salmonella.
  • 10. Host Defense Peptides Immune Stimulation  HDPs have a strong capacity to modulate the innate immune response by inducing chemotaxis and activation of various types of leukocytes.
  • 11. Host Defense Peptides  Because of these pleiotropic effects, HDPs have been actively explored as a new class of therapeutic agents against antibiotic-resistant microbes and other inflammatory diseases.
  • 12. Contents  Host Defense Peptides HDPs  Host Defense Peptides HDP and Sodium Butyrate  Trophic Effect Of Sodium Butyrate  Antibacterial Effect Of Sodium Butyrate  Other effects Of Sodium Butyrate
  • 13. In Vivo Induction Of The Avbd9 Gene Expression In The Intestinal Tract Of Chickens By Butyrate
  • 14. Sodium Butyrate and HDPs  Butyrate, a major species of short-chain fatty acids produced by bacterial fermentation of undigested carbohydrates in the intestine, was recently found to be capable of inducing HDP expression in humans, rabbits and poultry.
  • 15. Study Design Birds profile: Two-day-old male Cornish Rock broilers Treatment Feed with standard ration with or without supplementation of 0.1% and 0.2% butyrate for 2 days. Evaluation: Crop, cecal tonsil and cecum were collected from each bird and the AvBD9 gene expression was evaluated by real-time PCR.
  • 16.
  • 17. Reduction of the S. enteritidis titer in the cecal contents of chickens following oral supplementation of butyrate
  • 18. Study Objective  To examine the effect of supplementing butyrate in the feed on the titer of Salmonella enteritidis in the cecum following experimental infections.
  • 19. Study Design Birds profile: 20-day old male broilers were equally divided into two groups, each of 10. Treatment Feed with a standard antibiotic-free diet mixed with and without 0.2% sodium butyrate for 2 days.
  • 20. Cont. … Challenge Birds were then inoculated with 1X10 log 6 CFU of S. enteritidis phage type 13a and continued with butyrate feeding for another 4 days. Evaluation The S. enteritidis titer in the cecal content was quantitated from each bird by serial plating on Brilliant Green agar plates containing 20 mg/ml nalidixic acid.
  • 21.
  • 22. Study Design  This trial was similarly conducted with an additional group of 10 broilers fed with 0.1% butyrate.  Each dot represents the bacterial titer from a bird and the solid line represents the median value of each treatment.  Brackets indicate the statistical significance of differences
  • 23.
  • 24. Contents  Host Defense Peptides HDPs  Host Defense Peptides HDP and Sodium Butyrate  Trophic Effect Of Sodium Butyrate  Antibacterial Effect Of Sodium Butyrate  Other effects Of Sodium Butyrate
  • 25. Trophic Effect Of Sodium Butyrate  Sodium butyrate is a preferred source of energy for the enterocytes.  This will result in: 1. Better development of the intestinal villi. 2. Stronger gut lining.
  • 26. Study Design Trial Objective To evaluate the influence of sodium butyrate on digestibility of a corn-soya diet, as well as villi development. Birds Profile A total of 750 Cobb 400 one day old broiler chickens. Treatments
  • 27. Cont. … The trial evaluated 3 treatments: 1. Control (no additives) 2. 0.5 kg/t of Product X (92% sodium butyrate) 3. 1 kg/t of Product X. Evaluations 1. The digestible energy of the diet 2. The digestibility of the protein 3. The lengths and widths of the villis
  • 28. Digestibility of Energy Energy kcal/kg 3,105 3,264 3,285 Digestibility of Energy Control Product X 0.5g/kg Product X 1.0g/kg
  • 29. Protein Digestibility Protein Digestability 65% 66% 68% Protein Digestibility Control Product X 0.5g/kg Product X 1.0g/kg
  • 30. Villi Length 21 Days Old mm Villi Length 21 Days Old mm 0.98 1.52 1.24 Villi Length 21 Days Old mm Control Product X 0.5g/kg Gustor 1.0g/kg
  • 31. Villi Length 42 Days Old mm Villi Length 42 Days Old mm 0.94 1.31 1.12 Villi Length 42 Days Old mm Control Product X 0.5g/kg Gustor 1.0g/kg
  • 32. Villi Width 21 Days Old µm Villi Width 21 Days Old µm 224.70 260.80 197.70 Villi Width 21 Days Old µm Control Product X 0.5g/kg Gustor 1.0g/kg
  • 33. Villi Width 42 Days Old µm Villi Width 21 Days Old µm 138.50 314.20 317.20 Villi Width 42 Days Old µm Control Product X 0.5g/kg Gustor 1.0g/kg
  • 34. Conclusion  The digestible energy of the diet and the digestibility of the protein were significantly improved by the addition of sodium butyrate.  The lengths and widths of the villi were also positively affected by the addition of butyrate in the diet.  From this trial and previous research, it was concluded that inclusion of butyrate in the diet improves the digestibility of energy and protein by increasing intestinal absorption surface
  • 35. Contents  Host defense peptides HDPs  Host defense peptides HDP and sodium butyrate  Trophic effect of sodium butyrate  Antibacterial effect of sodium butyrate  Other effects of sodium butyrate
  • 36. VFA Antibacterial Effect  It has been demonstrated that VFAs can inhibit the growth of bacteria of the group of Enterobacteriaceae (Salmonella, Escherichia coli …). Mechanism of action  This is because the undissociated form of these acids can freely diffuse across the bacterial membrane.  Once inside the cytoplasm of the bacteria, the acid dissociates, thus releasing free hydrogen ion and reducing the pH, which causes internal cell damage.
  • 37. Butyric Acid  In this respect, butyric acid performs very well.  Hume et al. (1993) showed that butyric acid has a higher diffusion coefficient than other acids with a shorter chain, which allows it to pass through the bacterial membrane more easily.  Galfi and Neogradi (1995) found that the concentration of butyrate required to reduce the growth of E. coli by 50% is much lower than the concentration of the other volatile fatty acids, acetate and propionate.
  • 38. Butyric Acid Vs Other Fatty Acids Van Immerseel (2004) observed that butyric acid was the most effective acid to help controlling Salmonella enteritidis in broilers after a challenge. Number of colony forming units (CFU) per gram of ceca at day 8 after hatch of chickens orally inoculated with 1,000 CFU Salmonella enteritidis at days 5 and 6 and given feed supplemented with different organic acids (Adapted from Van Immersel, 2004)
  • 39. Butyric Acid Selective Action  It should be also noted that the antibacterial effect of butyrate is selective.  For instance, in 1990, Galfi and Bokori demonstrated that butyric acid favors the development of lactic flora which competes against the enteropathogenic one (Clostridia, E. coli, Coccidiae…).
  • 40. Butyric Acid Coccidia  Leeson et al. (2005) also observed that birds receiving butyric acid before coccidial oocyte challenge showed higher weight gain following the challenge compared with birds that received a control feed.
  • 41. Butyric Acid Salmonella  One very unique feature of butyric acid is its ability to negatively affect the virulence of pathogenic bacteria such as Salmonella.
  • 42. Cont. …  To infect an animal host, Salmonella must penetrate the intestinal epithelial barrier.  This very important step is called invasive phase and requires specific genes inside the bacterium.  The invasion genes are located on the so-called “pathogenicity island” and are encoding some regulatory proteins, some structural components of a needle complex, plus some effector proteins, which make possible the entry of Salmonella into the epithelial cells.
  • 43. Cont. …  The pathogenicity island itself is activated by another protein called HilA, which can be environmentally regulated.  Different publications have shown that butyrate and propionate can reduce the pathogenicity of Salmonella Typhimurium or Salmonella Enteritidis, whereas other acids such as acetic or formic are actually promoting the invasion of epithelium cells by the bacteria (Durant et al., 2000 - Van Immerseel et al., 2003 - Lawhon et al., 2005 - Gantois et al., 2006 - Huang et al., 2008).
  • 44. Cont. …  The exact mode of action of butyrate in the bacterial cell is still unknown but it seems it can interfere with the genetic expression and results in a down regulation of the pathogenicity island, regulating the invasive phenotypes of Salmonella.
  • 45. Contents  Host defense peptides HDPs  Host defense peptides HDP and sodium butyrate  Trophic effect of sodium butyrate  Antibacterial effect of sodium butyrate  Other effects of sodium butyrate
  • 46. Other effects Of Sodium Butyrate 1. Stimulation of Pancreatic Secretions  Although other organic acids also produced this effect, butyrate seems to be the one that causes the greatest stimulation of pancreatic secretions.  Katoh and Tsudo (1984) observed that the injection of sodium butyrate in pigs resulted in greater stimulation of pancreatic fluid secretion and increased amylase secretion.
  • 47. Cont. …  Katoh et al. (1989) demonstrated that intravenous injection of sodium butyrate in calves resulted in a higher secretion of pancreatic juice and greater release of proteins (particularly amylase) when compared to acetate and propionate.
  • 48. Cont. … 2. Stimulation of Insulin Secretions  Sano et al. (1995) also observed that the administration of butyrate caused an increase in the plasmatic levels of insulin, which normally results in higher amount of energy and proteins deposited in the animal tissues.
  • 49. Cont. … 3. Improves Absorption Of Electrolytes  Sodium butyrate also improves the absorption of electrolytes and reduces the incidence of diarrhoea (Galfi, 1989).

Editor's Notes

  1. Defensins are categorized by the presence of six conserved cysteine residues in the C-terminal mature sequence. Cathelicidins consist of a conserved cathelin domain in the pro-sequence with a highly diversified C-terminal mature sequence.
  2. All AvBDs are densely clustered on chicken chromosome 3q, whereas cathelicidin genes are located on chromosome 2p.
  3. pleiotropic عديدة المظاهر
  4. Trophic ذو علاقة بالغذاء