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Alternatives to Alternatives to thethe UseUse of of AntibioticAntibioticGrowthGrowth Promotores in RuminantsPromotores in Ruminants
S. CalsamigliaS. CalsamigliaDept. CiDept. Cièència Animal i dels Alimentsncia Animal i dels AlimentsUniversitat Autònoma de BarcelonaUniversitat Autònoma de Barcelona
sergio calsamigliasergio.calsamiglia@@uab esuab.es
FIBER STARCH
1 GLUCOSE
2 PYRUVATE
2 LACTATE
2 PROPIONATE
2 ACETATE
1 BUTYRATE
2 METHANE
+ 2 XH2
+ 2 XH2
- 4 XH2
- 2 XH2
- 2 XH2
- 2 XH2
N N MetabolismMetabolism in in thethe RumenRumen
CP
CP-Deg
CP-NoDeg
CP-Bac
NH3
AA
CP-InDig
UREA
Used Protein(15-30%)
Rumen
Liver
E
Req
MonensinMonensin⇓⇓ GramGram ++⇑⇑ GramGram --
⇓⇓ CatabolismCatabolism PepPep & AA& AA
⇓⇓ NHNH33
⇑⇑ PropionatePropionate⇓⇓ AcetateAcetate⇓⇓ ButyrateButyrate
⇓ CH4
Ban Ban ofof antibioticsantibiotics 2006 Regulation 1831/2003 ⇒⇒⇒⇒ searchsearch ofof alternativesalternatives
Objectives
Enhance Microbial Activities
Inhibit Microbial Activities
Fermentation and inefficiencies
Modulation
•Yeast/Fungi
0
1
2
3
4
5
6
Lact
ate
Util
izat
ion,
m
mol
/mg/
min
0 x1 x2.5 x5 x10Dose
Live SC SC Filtrate
Lactate Utilization by S. ruminantiumin Live vs Dead S. cerevesiae
Nisbet & Martin (1991)
0
1
2
3
4
5
6
7
Bac
teri
a C
ount
s (1
08 )
CTRL (102) YC-1 (103) YC-2 (108) YC-3 (146) YC-4 (189)
Strain (O2, nm/min/ml)
Total Cellulolytic
YC Strain, Oxygen Consumptionand Microbial Counts
Newbold et al. (1996)* (P<0,05)
* *
**
Live Yeast• Summary of 25 studies
-60
-50
-40
-30
-20
-10
0
10
20
30
40
50
60
PV
IMS
Pro
d G P L
MS
FND PB
CL
HCL
Tota
l
Celu
lo
Pro
teo
Pro
tz pH
Tota
l
Ace Pro
A:P Bu
Ram
if.
Lact
ato
NH3
Variación resp
ect
o a
l gru
po c
ontrol (%
)
9 18 8 8 8 3 10 14 5 1 1 0 1 0 4 17 14 16 15 10 15 7 6 14
Milk Digestibility
Microbiology
N=
FermentationProfile
-60
-50
-40
-30
-20
-10
0
10
20
30
40
50
60
PV
IMS
Pro
d G P L
MS
FND PB
CL
HCL
Tota
l
Celu
lo
Pro
teo
Pro
tz pH
Tota
l
Ace Pro
A:P Bu
Ram
if.
Lact
ato
NH3
Variaci
ón r
esp
ect
o a
l gru
po c
ontr
ol (%
)
Fermentationprofile
Microbiology
DigestibilityMilk
N= 14 17 11 11 11 8 7 6 6 0 2 2 3 1 3 9 8 9 9 8 9 8 1 9
Yeast Culture• Summary of 21 studies
Yeast/Fungi
Elimination of Oxygen
Estimulation fibrolytic bacteria
Estimulation oflactic acid use
Increase in fiber digestion
Increase in DM intakeand nutrient supply
Increase in production
Nutrients
EffectsEffects ofof S. S. cerevisaecerevisae andand A. A. oryzaeoryzae ononPerformance Performance ((YoonYoon & Stern, 1995)& Stern, 1995)
S. cerevisae: Summary 12 studies:– Average increase in fat corrected milk (4%): 0.58 L– Average increase in DM intake: 0.32 kg– The increase in DM intake justifies the improvement in
milk production.
A. Oryzae: Summary of 14 studies– Average increase in fat corrected milk (4%): 0.79 L– Average increase in DM intake: 0.36 kg– The increase in DM intake justifies the improvement in
milk production.
EffectsEffects ofof livelive vsvs culture culture ofof S. S. cerevisaecerevisae (Robinson & (Robinson & ErasmusErasmus, 2008), 2008)
Live: – DM Intake: +4.3%– Milk yield: 2.7%– Milk energy output: +3.4%
Culture– DM Intake: +1.8%– Milk yield: 3.6%– Milk energy output: +3.0%
Direct Fed Microbials: Energy
Fiber Starch
Glucose
AcetateMethane
Propionate
Lactate
Butyrate
SC/AO
SC/AO
Objectives
Enhance Microbial Activities
Inhibit Microbial Activities
Fermentation and inefficiencies
Modulation
•Yeast/Fungi•Plant extracts•Antibodies
Borcher et al., 1965; Oh et al., 1967
Oh et al., 1968; Nagy & Tengerdy, 1968
1960
1980
1990Ismael et al., 1990; Sivropoulou et al., 1995Sivropoulou et al., 1996; Helander et al., 1998
1970
2000
EssentialEssential OilsOils
Legal use of Monensin (1971-1975)
Ban on Antibiotic Use Anounced
Broderick & Balthrop, 1979
2000-2012: More than 65 papers and 4 reviews
16.0 †
2.5
6.4
CIN
5.55.46.8LPep N
17.0†13.0*19.2Ammonia N
4.6*4.4†1.9SPep + AA N
GARMONCTR
* P < 0.05 † P < 0.10
NitrogenNitrogen MetabolismMetabolism
Busquet et al., 2005a
NitrogenNitrogen MetabolismMetabolism
6.94.85.04.36.14.53.8LPep N
6.47.18.58.07.48.77.5Ammonia N
2.84.04.74.55.05.05.2SPep + AA N
CLOGINDILTEACADFENCTR
*
†
* P < 0.05 † P < 0.10 Busquet et al., 2005c
1* 2 3 4 5 6 7 8 9 10 11 12 13 14
Fluo
resc
ence
Time (seconds)
0
1
2
3
4
5
6
7
8
9
10
11
35 37 39 41 43 45 47 49 51 53 55
CTR1 P2 restrikcija HaeIII CIN1 P2 restrikcija HaeIII PCR GAR1 P2 neredcenCTR 1 (P2) HaeIII digestion CIN 1 (P2) HaeIII digestion GAR 1 (P2) undigested PCR product
CTR CIN
P. rP. rP. b
P. b
Mechanism of Action: (Prevotella)
(Ferme et al, 2004)
DietaryDietaryProteinProteinProteinProtein
PeptidesPeptidesAAAA
NHNH33
UreaUrea
MicrobialMicrobial ProteinProtein
XX EUGCIN
EO
Essential Oils: Protein
0
1.25
2.5
CTR MON MONx10 CIN
Busquet et al., 2005 (J. Dairy Sci.)
AcetateAcetate toto PropionatePropionate RatioRatio
c
a
b
abc (P<0,05)
b
Molar Molar ProportionProportion ofof ButyrateButyrate
0
5
10
15
CTR MON MONx10 CIN
mol
/100
mol
Busquet et al., 2005a (J. Dairy Sci.)
c
bb
abc (P<0,05)
b
Fibre Starch
Pyruvate
Acetate
Methane
Propionate
Lactate
Butyrate
Glucose+4 H+
+4 H+
-8 H+
-4 H+
-2 H+
-2 H+
Hipothesis: Inhibition of Methanogenesis
Monensin
EO
X
Hydroxy-Methyl-Glutaryl-CoAGAR
Hipothesis: Inhibition of methanogenesis
Acetyl-CoA
Mevalonate
Isoprenoids
X HMG-CoAreductase
ProteinIsoprenilated
MethanogenicArchaea
Hipothesis: Inhibition of methanogenesis
The methanogenic Archaea are the onlyrumen microorganisms that require theisoprenoid untis in the cell membrane.
Garlic extract and their active components may be very selective agaist this group of microbes
Essential Oils: Energy
Fiber Starch
Glucose
AcetateMethane
Propionate
Lactate
Butyrate
EO
Beef: Fermentation Profile0.073.15
0.611.050.850.160.190.04
6.05152.9
52.9*30.5*12.14.1*1,7*0.25*
6.14154.5
53.9*29.812.14.0*
1,81†
0.29
6.07155.5
53.9*31.2*10.73.8*1.7*
0.27†
6.1155.1
56.826.711.64.52.10.33
pH VFA total, mMVFA individual,%
- Acetate- Propionate- Butyrate
BC-VFA, mMC2:C3
L-lactic, mM
CIE SEMCAPANICTR
* P< 0.05; † P< 0.10 UAB
Beef: Dry Matter Intake
6,5
6,9
7,3
7,7
8,1
8,5
CTR ANI CAP CIE
Kg/
d
*†
* P< 0.05; † P< 0.10
9 %
5 %
UAB
Hypothesis
• How can we increase intake and maintain pH?
• Rumen fermentation, pH and DM intake patterns
PatternPattern of DMI and pHof DMI and pH
CAP
CTRL
(Rodriguez et al., un published)
ModifyingModifying thethe PatternPattern of DMI of DMI withwithAdditivesAdditives
CAP
CTR
(Rodriguez et al., unpublished)
Essential OilsEssential Oils
Antimicrobial activity: no search for the most antimicrobial, but for the optimal product/dose.
High diversity of substances (and effects).
Some have proven effects in vivo
Performance data limited, incosistent and urgently needed
Objectives
Enhance Microbial Activities
Inhibit Microbial Activities
Fermentation and inefficiencies
Modulation
•Yeast/Fungi•Plant extracts•Antibodies
InhibitionInhibition ofof SpecificSpecific MicrobialMicrobialGroupsGroups: : AntibodiesAntibodies
Immunization– Immunization Antibodies to the rumen
through saliva Reduction of specificmicrobial groups
Bacterial Bacterial CountsCounts afterafter ImmunizationImmunization
0123456789
S. bovis Lactobacilllus
Bac
teria
, (Lo
g10
CFU
/ml)
.
Control Immunized
Shu et al. (1999)
LactateLactate ConcentrationsConcentrations afterafterImmunizationImmunization
0
2
4
6
8
10
12
Control Immunized
Lact
ate
(nm
ol/L
)
L-Lactate D-Lactate
Shu et al. (1999)
InhibitionInhibition ofof SpecificSpecific MicrobialMicrobialGroupsGroups: : AntibodiesAntibodies
Immunization– Immunization Antibodies to the rumen
through saliva Reduction of specific microbial groups
Polyclonal antibodies– Immunization of chickens Polyclonal
antibodies Oral dose Reduction of specific microbial groups
Antibodies against S. bovis and pH:
Day 11=2.5kg concentrate/ hdDay 12=5kg concentrate/ hdDay 13=7.5kg concentrate/ hdDay 14=10kg concentrate/ hdDay 15=12.5kg concentrate/ hd
Days post PAP-Sb or CTRL feeding
(Blanch et al., 2006)
Antibodies and S. bovis and M. elsdenii
0,00
20,00
40,0060,00
80,00
100,00
120,00
140,00160,00
180,00
200,00
A1 A10 A P1 P4
ng/m
L R
F
CT
DAY: P < 0.10
0,0020,0040,0060,0080,00
100,00120,00140,00160,00180,00200,00
A1 A10 A P1 P4
ng/m
L R
F
S. bovis M. elsdeniiDAY: NS
(Blanch et al., 2006)
Effect of Effect of S.bS.b. and . and F.nF.n. Antibodies on . Antibodies on RuminalRuminal pHpH
5
5,2
5,4
5,6
5,8
6
6,2
None PAPSb FA Both
Treatment
Rum
en p
H
a
b
bb
a, b (P < 0.05). DiLorenzo et al. (2005)
Effect of Antibodies on Feedlot Effect of Antibodies on Feedlot PerformancePerformance
Initial BW, kgInitial BW, kg 265265 267 266 267267 266 267 0.720.72
Final BW, kgFinal BW, kg 539 545 544 543539 545 544 543 2.762.76
ADG, kg 1.70ADG, kg 1.70 1.74 1.74 1.741.74 1.73 0.021.73 0.02
DMI, kg/d 9.46DMI, kg/d 9.46 9.40 9.32 9.37 0.109.40 9.32 9.37 0.10
Gain/FeedGain/Feed 0.1790.179aa 0.1850.185bb 0.1860.186bb 0.1840.184abab 0.0020.002
CTR Fn CTR Fn SbSb Both SEBoth SE
a,b (P < 0.05) DiLorenzo et al. (2005)
Summary: Energy
Fiber Starch
Glucose
AcetateMethane
Propionate
Lactate
Butyrate
SC/AO
EO AB
SC/AO
The objectives of modulating ruminal fermentation should be clearly defined for each production system.
The careful selection and combination of additives with specific effects and synergies may help in optimising ruminal fermentation.
Data on benefits of these combinations is not available.
The use rumen modifiers can only be justified if animal performance and economic benefits are obtained. However, this information is very limited and urgently needed.
Conclusions
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