GLOBACID FOR POULTRY

GLOBACID Acidifier

Main acids used in animal nutrition

• Many acids are available, organic or minerals; liquids or solids • Mineral or inorganic doesn’t means that there are not naturaly essential for

organism

Acids Concentration % Physical presentationPropionic 99 LiquidButyric 99 LiquidCitric 99 SolidBenzoic 99 SolidSorbic 99 SolidFormic 85 LiquidLactic 80 LiquidAcetic 80 LiquidFumaric 99 SolidPhosphoric 75 LiquidHydrochloric 30 Liquid

GENERALITIES ON ACIDS

Acidity – Mineral and organic acids

• Main acids are divided into mineral and organic acids• Mineral acids :

• Hydrochloric HCl -> H+ + Cl-

• Sulfuric acid H2SO4 -> H+ + SO4-

• Phosphoric acid H3PO4 -> H+ + H2PO4-

• Organic acids or carboxylic acids :

• All molecula having a COOH (carboxylic) function :

• COOH -> H+ + COO-

• Formic, acetic, propionic, butyric, etc…

• It is simply related to the concentration of H+ or more precisely of H30+ in an aquaeous solution

• Consequently, any molecula able to liberate in solution is H+ an acid

STRONG AND WEAK ACIDS

• Weak acids :

• The dissociation is not total , balance between 2 forms, one called dissociated, the second called non dissociated :

AH <-> H+ + A-

• Like any chemical balance this equilibria allways respect stricly a chemical rule due to the equilibrium constancy K

• K = [H+][A-]/[AH]

• we speak about Ka, K for constancy a for acidity

• The higher the constancy, the more H+ is produced, so the stronger is the acid

• pKa is equal to -LOG10 of Ka, the lower the pKa, the stronger the acid

• Strong acids :

• The dissociation in solution is allways complete and no reversible

• AH -> H+ + A-

STRONG AND WEAK ACIDS IN ANIMAL NURITION

• Acid classification :

• Organic acids are allways weak acids :

• All organic acids have a pKa and consequently are weak acids

• Some minerals acids are strong acids :

• For example HCl is a strong acid, allways completly dissociated.

• Even if this acid that can be found in the organism, especially in the stomach. It is very difficult to use it in animal production.

• The risk of over dosage is important.

• In the organisme, its production is strongly regulated according to the pH of the stomach, so no risk of excess

STRONG AND WEAK ACIDS IN ANIMAL NURITION

• Acid classification :

• Some mineral acids are also weak acids :

• 2 of them are fundamental for the regulation of pH in the organism :

• Phosphoric acid :

• H3PO4 H+ + H2PO4- pKa12.2

• H2PO4- H+ + HPO4-- pKa7.3

• The second acidity is the main important in the regulation of intracellular pH and urine pH

• Carbonique acide :

• H2CO3 H+ + HCO3- pKa = 6.3

• This reaction is fundamental for blood, intestinal environmental pH balance

• So mineral acids in some case means also physiological and even fundamental for organism metabolism regulation

Pka effect on pH

• H30+ = c Ka1 + c Ka2 + c Ka3 so pH depends on concentration and pKa only

• Only the lower pKa has an effect on pH for poly acids• + 1 point of pKa -> + 10 times more concentration

needed to reach the same pH (0.1 x 10-2 = 1.0 x 10-3)• Lower pH are obtained with lower pKa

pKa Concentration mol/l pH Concentration mol/l pH0 0.1 1 1.0 02 0.1 1.5 1.0 13 0.1 2 1.0 1.54 0.1 2.5 1.0 25 0.1 3 1.0 2.5

Acid pKa1 pKa2 pKa3Hydrochloric 0Phosphoric 2.15 7.09 12.32Fumaric 3.03 4.54Citric 3.13 4.76 6.4Formic 3.75Lactic 3.83Benzoic 4.19Acetic 4.74Sorbic 4.76Butyric 4.8Propionic 4.87

Strengh of acids

• Conclusion : lower pKa makes lower pH

More efficient to reduce pH

Number of acids function concentration

Acid Concentration % Molecular weight Number/H+ moles/KgFormic 85 46.0 1 18.5Fumaric 99 116.1 2 17.1Citric 99 192.1 3 15.5Propionic 99 74.1 1 13.4Acetic 80 60.1 1 13.3Phosphoric 75 98.0 1.5 11.5Butyric 99 88.1 1 11.2Lactic 80 90.1 1 8.9Sorbic 99 112.1 1 8.8Hydrochloric 30 36.5 1 8.2Benzoic 99 122.1 1 8.1

Formic can be the more concentrated liquid acid

Use to be a cheap by-product so used in animal nutrition

But it is highly corrosive and non palatable

ACIDS FOR pH REDUCTIONBACTERIOSTATIC EFFECT

STAPHYLOCOQUES

Non mutiplication area

4 5 6 7 83

E. COLI

CLOSTRIDIUMSALMONELLES

pH

4,4 6 8

7,8

6,84,2

5

Multiplication area

Bacteriostatic action throught pH reduction

Protein digestion and pH

Pepsine activity and consequently stomach protein digestion is optimum when pH is low

Reducing pH in stomach also improves protein digestibility

Enzym activity

BACTERICID EFFECT OF ACIDS

Acids forms and pH

Ka = [H3O+] [A-] is a constant value

AH undissociated form dominant

A- dissociated form dominant

pKa pH

Same proportion

[AH]

If [H3O+] increases [AH] must increase or [A-] must decrease

• When pH is low, acid conditions AH is the most important

• When pH is higher than pKa, A- is the most important

pH = pKa or [H3O+]=[A] [AH]=[A]

pH effect on acid dissociation

pH 1.8 2.8 3.8 4.8 5.8 6.8 7.8 8.8 9.8 10.8 11.8

BuCOOH 99.9 99 90 50 10 1 0.1 0.01 0.001 0.0001 0.00001

BuCOO- 0.1 1 10 50 90 99 99.9 99.99 99.999 99.9999 100

pH 1.8 2.8 3.8 4.8 5.8 6.8 7.8 8.8 9.8 10.8 11.8

HCOOH 99 90 50 10 1 0.1 0.01 0.001 0.0001 0.00001 0.000001

HCOO- 1 10 50 90 99 99.9 99.99 99.999 99.9999 100 100

The lower the pH, the more undissociated form,

ex : 90% at pH 3.8 vs 50% at pH 4.8 for butyric

The higher the pKa, the more undissociated form at same pH,

ex : at pH 4.8, 50% for butyric vs 10% for formic

Butyric acid

Formic acid

BUFFERING CAPACITY• Can be measured at pH 3, 4 or 5• For the stomach activity, the pH 3 is the most important

(pepsine activity)• The liberation of acidity before the reference pH is important. • At pH 3, formic or lactic acid with a pKa of 3.8% have only

15% of efficiency, while phosphoric and citric respectively 88% and 43%

Acidity release according to pH

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1 2 3 4 5 6 7pH

mole

s H

+

Formique 85

Lactique feed 80

Propionique 99

Phosphorique 75

Butyrique

Citrique poudre 99

Sorbique 99

RCOOHIn acid environment

pH

H+

pH

H+

H+RCOO-

BACTERICIDE ACTION OF ACIDIFIER

BACTERIANeutral environment

RCOO- polar,can’t leave the cell

RCOO-

Accumlates in the cell, cause of RNA dammage

1st cause of cell death

H+ acidify bacteria content

To increase pH, bacterias must excrete H+, spending energy

Energy

2nd cause of cell death

RCOOHIn neutral environment

RCOO- polar,can’t enter the cell

No bactericid action

INHIBITION OF COLIBACILLI BY VFA

• BUTYRATE > PROPIONATE > ACETATE• pH reduction has a synergistic effect

pH=7.0pH=6.6

pH=6.2pH=5.8

n-Butyrate

Propionate

Acetate

0

20

40

60

80

100

120

140

160

mM

/l

• Gàlfi and Neogràdy, 1992

VFA concentration to reduce by 50% Colibacilli growth

200000

17000

1

10

100

1000

10000

100000

1000000

CTR Globacid

Globacid LF-60-Plus : Effect against E. Coli in the intestine of broilers

Log (Nbr/gr)

Regional Animal Health Institute - Torhout (B) : Dr. R. Wyffels - 15 days trial period - 5 days pretrial period - 4 kg/ton

SYNERGIE BETWEEN ORGANIC ACIDS AND

PHOSPHORIC ACID

Synergy phosphoric and organic acids

Bactericide action of Acids mix

pH3.8 pKa

R-COOH R-COO-

Lowering the pH, phosphoric acids makes the undissociated form dominant

When acids will react with other components, the strongest acid in a mix is allways the first one to react. So phosphoric acid reacts allways before organic acids in the mix, preserving their efficiencyOrganic acids can have a more bactericide effect

H3PO4 acid effect R-COO-R-COOH

pH reduction and phosphorique inclusion

2.0

2.2

2.4

2.6

2.8

3.0

3.2

3.4

100%

for

mic

100%

lacti

c

L+P 10

L+P 20

L+P 30

L+P 40

L+P 50

L+P 60

L+P 70

L+P 80

L+P 90

100%

Pho

spho

ric

Already 20% of phosphorique makes less 10% of lactique inactive

OPCL = 50% compromise with % efficiency and pH reduction (protein digestibility)

22%

11%

9%7%

6% 6% 5% 5% 4%

2.5 Kg inclusion /T18%

% of dissociated inactive form

Consequences

• Organic acids such as Formic acid and lactic are the more efficient to have a bactericid effect

FormicLactic

In same quantities, Formic as a smaller molecular weight so can bring more acid per kg

But as a smaller molecula, its absorption is also quicker in the stomach

Consequences

• Organic acids associations such as Phosphoric/Lactic are as efficient to have a bactericid effect

Formic

Phosphoric

Lactic

DIGESTIVE TRACT ANATOMY AND PHYSIOLOGY

Physiology of poultry digestive tract

Crop

pH 5.5

ProventriculuspH 2.5 – 3.5

GizzardpH 1.5 – 3.5

Pancreas

JejunumpH 6.5-7

CæcumColon-Rectum

CloacapH 8

pH: Normal pH in digestive tract

CæcumpH 6.9

DuodenumpH 5 - 6

Foie

Log 10 Crop Gizzard Duodenum Ileum Caeca

Lactobacilli 8.7 7.3 8.0 8.6 8.7

Enterococus 4.0 3.7 4.0 4.2 6.7

Coliformes 1.7 - 2.0 2.7 5.6

Yeast 2.7 - 1.7 - 2.0

Clostridia - - (-) (-) 9.0

Anaerobes - - - - 10.0

Streptococus anaerobic - - - - 10.0

IleumpH 7 – 7.5

Bacterial flora is highly developped all along the digestive tract

Acidification naturally exists in the first part of the digestive tract

Antibiotics growth promotors are used to control this flora

Physiology of poultry digestive tract

Mouth

Feed are almost no fragmentated

Ptyaline* action begins and continu in crop(Surdeau et Hénaff, 1979)

* Ptyaline: enzyme (amylase) degrades starch

Natural presence of

8.710/g Lactobacille

s

Physiology of poultry digestive tract

Crop

Important production of mucus to humidify feed

Feed reserve playing important role in feed ingestion

pH 4,5 – 6,5

Emptying of crop is regulated by particule size and quantity (quicker for meal)

pH variation is related to feed quantity. pH reduction is due only to fermentation because there are no acidic secretion

The flora is composed mainly of lactobacille (Gabriel Irène et al, JRA 2003) that are responsible for lactic fermentations (Wielen et al, 2000)

Acidification interest

• In the crop, no acid secretion • Acidifier allows to accelerate the pHJ reduction and favorise

the development of lactobacille flora and is detrrimental for salmonella or other pathogen development

The vertical transmission of salmonellas and formic acid treatment of chicken feed. A possible strategy for control.

Humphrey TJ, Lanning DG.

Public Health Laboratory, Heavitree, Exeter, UK.

The treatment of feed given to laying hens with 0.5% formic acid reduced significantly the isolation rate of salmonellas and was associated with a reduction in the incidence of infection in newly hatched chicks. These improvements were not sustained until slaughter, however, as growing birds acquired salmonellas, probably from feed which was not acid treated. The data indicate that formic acid treatment of chicken food could have important benefits for the public healthEpidemiol Infect. 1988 Feb;100(1):43-9.

Physiology of poultry digestive tract

Proventriculus

Secrete high amount of hydrochloric acid

pH arounnd-pH 1,5 – 4,5-

pH 1,5 – 4,5

Pepsinogene transformation in pepsine is not complete

Proteolyse only begins

Content stays for 10 to 60 mn before entering in the gizzard

Proventriculeus

Physiology of poultry digestive tract

Gizzard

pH is low - pH 1,5 à 3

Proteolys is important under pepsine action

pH 1,5 – 3

Mecanical activity og gizzard is important, but lower for meal feedThe low pH allows to solubilize minerals (layer must solubilose 7 to 8 g of calcium carbonate per day)

Gizzard

Physiology of poultry digestive tract

Duodenum

Junction between gizzard and duodenum is recovered by a mucis to protect from high acidity

Increase pH content to 6 – 7 by endogenous secretion of sodium bicarbonate

pH 6 – 7

Duodenum

Bacterial growth is controled by the growth of high amount of enzymes, bile salts (antibacterial) and oxygen.

Physiology of poultry digestive tract

Jejunum and ileum

Place of chemical digestion by intestinal and pancreatic enzymes

Villositary absorption of nutriments, water and minerals

Jejunum and ileum

Digestive flora can be located in the lumen or attached in the intestinal mucus

pH 6,5 – 7,5

Physiology of poultry digestive tract

Intestinal villosities

Digestive flora depends of nutriments, transit speed presence or not of pathogen bacteria

Physiology of poultry digestive tract

Caecas

Place of bacterial fermentation, riche on anaerobic bacteria, producing AGV, acetic, propionic, butyric, formic (Mead, 2000)

Caecal contents are renewd 1 to 2 times per dayUrina is deversed to produce feces

Presence of more 1010 anaerobes

Poultry bacterial flora is stable at 2 weeks of age at intestinal level. It is required 4 to 6 weeks for the caecal flora to stabilise (Gabriel Irène et al, Inra 2003)

pH 6,5 – 7,5

Those organic acid have antibacterial effects (Vielen et al, 2000)

ACIDIFIERS AND POULTRY PERFORMANCE

Globacid LFPAGlobacid LFPA : : SalmonellaSalmonella

inhibitor in feedinhibitor in feed

1

10

100

1000

10000

100000

1000000

10000000

0 4 8 12 16 20 24Hours

Log (cfu/gr)

Globacid LFPA P

Control

Marivi Gnilo Colle : BS-Thesis at UPLB - April 2000 pH= 7 / moisture =12%

1

10

100

1000

10000

100000

1000000

0 4 8 12 16 20 24

pH = 5 pH = 6 pH = 7 pH = 8Log (cfu/gr)

Hours

Globacid LFPA-60 : :Globacid LFPA-60 : : anti- anti-Salmonella Salmonella activity in relation to activity in relation to feed pHfeed pH

Marivi Gnilo Colle : BS-Thesis at UPLB - April 2000 moisture =12 %

Globacid LFPA : Globacid LFPA : anti- anti-Salmonella Salmonella activity in relation to activity in relation to

feed moisture contentfeed moisture content

1

10

100

1000

10000

100000

1000000

10000000

100000000

0 4 8 12 16 20 24Hours

Log (cfu/gr)

M = 10% M = 12 % M = 14 % M = 16 %

Mark Kristoffer Ungos Pasayan : BS-Thesis at UPLB - April 2000 pH= 6

Mortality after contact with infected chicken by Salmonella Mortality after contact with infected chicken by Salmonella gallinarumgallinarum

Control Propionic+Formic

Mortality 76% 33%

• Organic acid helps to control pathogen bacteria, and patholgy incidences

Berchieri and Barrow, 1996

Crop pH Salmonella Log10

CONTROL 5.96 4.17

0.47% LACTIC ACID 5.47 2.17

Avila et al, 2003

Lactic acid effect on crop pH and salmonella controlLactic acid effect on crop pH and salmonella control

Comparison of some growth promoters in turkey feed AFSAA trial

Control Avilamycin MOS Organic acid

Weight 28d        

1st trial 1013 1068 1062 1030

2nd trial 981 1024 997 1043

• Organic acid can provide similar zootechnical performance as antibiotics growth promoters

• Organic acids highly improves litter quality

• Constant in most of the acidified feeds

Comparison of some growth promoters in turkey feed AFSAA trial

55

57

59

61

63

65

67

69

71

73

75

Control Avilamycin MOS Organic acid

Lit

ter

dry

mat

ter

Layers hen (repro)

2 buildings of 6000 animals (1 control, 1 acidifier)

Egg weight improvement

Layers hen (repro)

2 buildings of 6000 animals (1 control, 1 acidifier)

% Rejected eggs

Layer - Japan

Number of eggs + 6,8 %

Broken eggs - 64 %

FC - 8 %

Feces weight - 11,7 %

Moisture feces - 3 %

Performance index + 6 %

TOTAL BACTERIAL CONTROL PROGRAM

In the first part : Globacid = acidification action

In second part of the digestive tract : Globamax = Calcium Butyrate Action

All along the intestine for strong antisalmonella treatment : Globatan

Globacid

Wide action all along the digestive tract

Globamax

Globamax

Globatan

Globatan