Use of MIR spectrometry to qualify
milk of Normande cows
Focus on fine milk composition from results of
PhénoFinlait program
Marine Gelé
Marine.gele@idele.fr
15th September 2014
www.idele.fr
www,idele,fr
Normande breed international conference, Le Pin-au-Haras
Presentation outline
1. PhénoFinlait : general information and advances for
dairy sector
2. Normande cows’ milk: observations from
PhénoFinlait
 Milk yield and milk composition
 Focus on fine milk composition



Fatty acids, proteins
Genetic parameters
Influence of physiological, nutritional and seasonal factors
3. Calcium content in milk : first results
4. MIR spectrometry, method to phenotype new traits
www.idele.fr
Presentation outline
1. PhénoFinlait : general information and advances for
dairy sector
2. Normande cows’ milk: observations from
PhénoFinlait
 Milk yield and milk composition
 Focus on fine milk composition



Fatty acids, proteins
Genetic parameters
Influence of physiological, nutritional and seasonal factors
3. Calcium content in milk : first results
4. MIR spectrometry, method to phenotype new traits
www.idele.fr
PhénoFinlait
« PHENOTYPING » and « GENOTYPING » in the main cow, goat
and sheep populations to improve the knowledge and the
management of fine milk composition (fatty acids and proteins)
National R&D program
Cow, goat and sheep dairy sectors
Multidisciplinary program
Common interests between the dairy sector actors
(industry, farming, research, development…)
www.idele.fr
A lot of varied actors
Cniel
France Génétique Elevage
UNCEIA, ANIO et Capgènes
FCEL et CNBL
Inra
Actalia
Institut de l'Elevage
And on field …
•
•
•
•
A national
program
Milk analysis
laboratories
Breeding
companies
Cows
Ewes
Goats
Goat-Ewe
Milk recording organizations in 26 departments
10 breeding companies and insemination centres
9 milk analysis laboratories + Labogéna
And 1500 farmers !
www.idele.fr
Cow-Goat
Data collection
88 274 cows, 33 710 ewes, 71 551 goats
In 1 540 farms de 26 departments
Selection of 12000 cows, 4000
ewes, 4000 goats
MRO
Breeding companies
Milk
Survey :
Feeding system…
Blood
Freezing
Milk analysis
laboratory
CRB-GADIE
Fatty acids,
Proteins
BD
PFL
Labogéna
Genotypes
Inra CTIG
Phenotyping
www.idele.fr
SNIGs
Genotyping
General schedule
N milk samples
1.Analysis
methods
Reference analysis
MIR spectrum
• FA: Gas chromatography
• Proteins : liquid chromatography / mass
spectrometry
Mathematic
model
Calibration equations
2.Constitution of
the database
3.Data
interpretation
Spectral database
On-farm
survey
FA and proteins
profiles
Research of
available levers in
farms
www.idele.fr
Genotypes
Research of
genetic
determinants
Presentation outline
1. PhénoFinlait : general information and advances for
dairy sector
2. Normande cows’ milk: observations from
PhénoFinlait
 Milk yield and milk composition
 Focus on fine milk composition



Fatty acids, proteins
Genetic parameters
Influence of physiological, nutritional and seasonal factors
3. Calcium content in milk : first results
4. MIR spectrometry, method to phenotype new traits
www.idele.fr
Milk production and
composition
Normande cows produce less milk but more fat
and protein content
Montbeliarde
Holstein
(n = 92 267)
(n = 198 378)
20.1 ± 6.4
Normande
CLO 2010
19.4
24.4 ± 6.9
26.8 ± 8.1
Fat content (g/kg)
43.6 ± 6.6
42.8
39.3 ± 5.5
39.7 ± 6.8
Protein content (g/kg)
35.4 ± 3.5
34.5
33.4 ± 3.2
32.8 ± 3.8
Urea (mg/L)
241 ± 88.8
282 ± 89.5
206 ± 113
Lactose (g/kg)
47.9 ± 2.8
48.5 ± 2.8
48.7 ± 2.8
Cells (103)
320 ± 770
231 ± 606
288 ± 730
Trait
Normande
Milk yield (kg/day)
(n = 117 593)
Official milk record
Results from program
www.idele.fr
240
FOCUS ON
fine milk composition
determined by MIR spectrometry
53 fatty acids, groups of FA or ratios of FA
7 proteins or groups of proteins
Estimated traits
Very good precision
(R²>0,91 and relative sy,x <5%)
Fatty acids
FC, C4:0, C6:0, C8:0, SFA, MUFA,
UFA, SCFA, MCFA, Sum C4+C6, even
FA from C4 to C10, , even FA from C4
to C12, , even FA from C4 to C14, FA
with 16 carbons or less, SFA with 16
carbons or less, even SFA with 16
carbons or less, Sum
C12:0+4C14:0+C16:0
Proteins
PC, Total of caseins, β casein
Results from program
www.idele.fr
Good precision (R²>0,82 and Correct precision (R²>0,70 and relative
relative sy,x between 5 and 10%)
sy,x between 10 and 15%)
C11:0, C15:0, C15:0 iso, C17:0, C18:0,
C20:0, C17:1 c10, C18:1 c15, C18:1 t12,
C18:1 t16, C18:2 c9 c12, PUFA,
C10:0, C12:0, C14:0, C16:0,
PUGA/SFA, trans FA, Total C18:1trans,
C18:1 c9, LCFA, Total C18:1, Total Total des C18:2, Total C18:2 omega 6,
C18:1cis, elongation index,
omega 6 FA, omega 7 FA, omega 7 cis
atherogenicity index
FA, odd FA from C5 to C11, odd FA from
C5 to C15, odd FA from C13 to C17,
C18:1/C16:0, 18:1c9/C16:0,
desaturation index for C16,
Sum α-lactalbumin + β-lactoglobulin, βαs1 casein, αs2 casein, κ casein
lactoglobulin
More SFA, especially
MCFA, in Normandy
cows’ milk fat
Trait (% of total FA)
Saturated Fatty Acids (SFA)
Butyric acid C4:0
Caproïc acid C6:0
Caprylic acid C8:0
Capriqc acid C10:0
Lauric acid C12:0
Myristic acid C14:0
Palmitic acid C16:0
Mono-Unsaturated Fatty Acids (MUFA)
Total C18:1
Total C18:1 cis
Oleic acid C18:1c9
Poly-Unsaturated Fatty Acids (PUFA)
Total C18:2
Estimated data by MIR with PhénoFinlait equations
Results from program
www.idele.fr
Montbeliarde
Normande
Holstein
(n = 91 784)
(n = 115 838)
(n = 194 530)
67.1 ± 5.3
3.73 ± 0.5
2.11 ± 0.3
1.30 ± 0.2
2.67 ± 0.6
3.09 ± 0.8
11.8 ± 1.7
28.0 ± 4.7
28.9 ± 4.7
25.5 ± 4.7
22.6 ± 4.2
21.6 ± 4.0
4.0 ± 0.7
2.76 ± 0.6
69.0 ± 5.1
3.77 ± 0.4
2.15 ± 0.2
1.34 ± 0.2
2.78 ± 0.6
3.20 ± 0.8
11.8 ± 1.7
28.0 ± 4.6
27.3 ± 4.7
24.1 ± 4.5
21.1 ± 4.1
20.6 ± 4.0
3.6 ± 0.6
2.51 ± 0.5
67.9 ± 5.2
3.68 ± 0.5
2.08 ± 0.3
1.29 ± 0.2
2.63 ± 0.6
3.09 ± 0.8
11.8 ± 1.5
28.1 ± 5.2
28.3 ± 4.8
25.0 ± 4.7
21.9 ± 4.1
21.3 ± 4.2
3.7 ± 0.6
2.60 ± 0.5
Genetic determinism
reflects fatty acids
synthesis mechanisms
Heritability
Moderated for SFA with short or medium chain,
which are synthetized in the udder
Lower for LCFA, mainly brought by food and fat
mobilization
Low coefficient of genetic variation (1 to 5%)
Genetic correlations:
FC correlated + with SFA and C16:0
FC correlated - - with MUFA and PUFA
Results from program
www.idele.fr
Trait
Heritability
FC (g/kg)
0.35
SFA
0.19
C4:0
0.48
C6:0
0.31
C8:0
0.32
C10:0
0.32
C12:0
0.34
C14:0
0.26
C16:0
0.19
C18:0
0.15
MUFA
0.21
C18:1
0.20
PUFA
0.21
C18:2c9t11
0.10
ω3
0.20
ω6
0.14
Similar protein composition
between breeds.
κ casein proportion slightly lower
and αs2 casein higher
In Normande milk
Trait (% of total PC)
Total caseins
αs1 casein
αs2 casein
β casein
κ casein
Sum α-lactalbumin + β-lactoglobulin
Estimated data by MIR with PhénoFinlait equations
Results from program
www.idele.fr
Montbeliarde
Normande
Holstein
(n = 91 469)
(n = 117 391)
(n = 129 509)
83.9 ± 1.4
28.0 ± 0.6
9.66 ± 0.3
36.3 ± 0.9
9.97 ± 0.6
12.4 ± 1.2
83.5 ± 1.2
27.8 ± 0.7
9.89 ± 0.3
36.2 ± 1.2
9.87 ± 0.5
11.9 ± 1.2
82.9 ± 1.4
27.9 ± 0.7
9.70 ± 0.4
36.1 ± 1.2
9.44 ± 0.6
12.5 ± 1.4
The origin of milk protein
composition variability is
essentially genetic
Heritability
Higher than fatty acids heritability
Very high for αs1 and κ caseins and βlactoglobulin
Low coefficient of genetic
variation (0,5 to 7%)
Trait
PC(g/kg)
αs1 casein
αs2 casein
β casein
κ casein
α-lactalbumin
β-lactoglobulin
Genetic correlations :
PC correlated more with caseins than
with whey proteins
www.idele.fr
Results from program
Heritability
0.41
0.57
0.25
0.39
0.55
0.53
0.72
SFA, total caseins and β
casein proportions increase
MUFA and other proteins
proportions decrease
DIFFERENCE WITH THE
MODALITY 8-14 DAYS
Caseins
DIFFERENCE WITH THE MODALITY 8-14 DAYS
At the beginning of lactation:
SFA
8-14
DAYS
MUFA
15-50
DAYS
PUFA
51-120 121-200 201-350 351-500
DAYS
DAYS
DAYS
DAYS
LACTATION STAGE
Results from program
8-14
DAYS
www.idele.fr
15-30
DAYS
31-60 61-90 90-180 181-270 271-350
DAYS DAYS DAYS DAYS DAYS
LACTATION STAGE
Parity influences
fatty acid profile:
Primiparous cows produce less SFA and more MUFA than multiparous
Multiparous of different lactation rank have similar profiles
… but does not influence protein profile
Difference with the primiparous
1st
2nd
3rd
+
+
SFA
MUFA
PUFA
Results from program
www.idele.fr
+
Caseins
4th
and more
When days are long:
DIFFERENCE WITH THE
DECEMBER TEST-DAYS
SFA, C16:0, C14:0 and αs1 casein proportions decrease
MUFA, C18:1c9 and β
SFA
MUFA
PUFA
casein proportions increase
DIFFERENCE WITH THE
DECEMBER TEST-DAYS
Caseins
DEC JAN FEBR
MAR
JULY SEPT
MAY JUNE
APRIL
AUG OCT
NOV
MONTH OF TEST DAY
NOV
DEC
JAN
FEBR
MAR
APRIL
MAY
JUNE
JULY
AUG
MONTH OF TEST DAY
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SEPT
OCT
Results from program
Main feeding systems
Grass silage
Barn dried hay
Floor dried hay
% in the diet
Pasture
Maize/sorghum silage
Grass silage
Floor dried hay
Barn dried hay
Energy concentrates
Nitrogen concentrates
Minerals and additives
Other
Maize silage + grass silage
www.idele.fr
Maize silage
Pasture + maize silage
Pasture
Feeding system influences
fatty acid profile:
• In relation to maize silage diet :
UFA proportions are slightly higher with grass silage and hay
Pasture significantly decrease SFA proportions and increase MUFA and
PUFA proportions
… but has a minor influence on protein profile
SFA
Barn dried hay
Floor dried hay
Grass silage
Maize silage + grass silage
Pasture + maize silage
Pasture
Caseins
MUFA
PUFA
Results from program
Difference with the maize silage
based diet
www.idele.fr
Difference with the maize silage
based diet
Presentation outline
1. PhénoFinlait : general information and advances for
dairy sector
2. Normandy cows’ milk: observations from
PhénoFinlait
 Milk yield and milk composition
 Focus on fine milk composition



Fatty acids, proteins
Genetic parameters
Influence of physiological, nutritional and seasonal factors
3. Calcium content in milk : first results
4. MIR spectrometry, method to phenotype new traits
www.idele.fr
Determining milk calcium content by
spectrometry MIR is possible
300 milk samples
Reference analysis:
Flame absorption
spectrometry
MIR spectrum
Mathematic
model
Calibration equation for milk calcium content
• R²=0.80 and relative error = 3.6%
 Equation applied on
PhénoFinlait spectral
database
Normande
Montbeliarde
Holstein
1315 ± 90
1239 ± 87
1220 ± 96
Results from M. Wald training period
www.idele.fr
First results in genetics
Heritability = 0.74
Coefficient of genetic variation = 4.3%
Genetic correlation:
Negative with milk yield (-0.18), cells and milking speed
(-0.31 to -0.34)
No correlation with morphology traits
Positive with FC and PC (+0,42 and +0,44), caseins (+0,28
to +0,42) and β-lactoglobulin (+0,34)
Results from M. Wald training period
www.idele.fr
Evolution of milk calcium content over a
lactation
C. Hurtaud1, M. Johan1, S. Leurent2, Y. Gallard2 et L. Delaby2
1INRA-Agrocampus Ouest UMR 1080 Production du Lait
F-35590 Saint-Gilles, France
2INRA, Domaine du Pin-au-Haras, F-61310 EXMES, France
.023
Objectives of the trial
Identify the factors which have an influence on the
calcium content of milk
By following 2 dairy cows breeds during the whole
lactation (Normande, NO and Holstein, HO)
Study of the effect of feeding
Study of the effect of season
Study of the effect of stage of lactation
24
Calcium content during the year
Normandy
Prim’Holstein
Significant
effects of the
stage of
lactation and of
the month
Month
25
Breed and feeding systems
Winter
HO
NO
Intensive Extensive Intensive Extensive
MY, kg/d
33.2
23.9
26.5
18.5
PC, g/kg
32.3
29.6
34.2
32.4
Ca total, mg/kg
1303
1242
1403
1362
During grazing period, no effect of the feeding system
Breed: NO cows + 86 mg/kg Ca total (P= 0.001)
26
Calcium content and stage of lactation
Calcium
content
decreases with
the stage of
lactation
80
70
exported Ca , g/d
60
50
y = -0,0625x + 37,78
R² = 0,3374
40
30
20
10
0
0
50
100
150
200
250
300
350
400
450
days of lactation
27
Calcium content and duration of the
day
Prim’Holstein
Calcium total, mg/kg
Normandy
Photopériod, h
Intensive
Extensive
Photoperiod
Long days
Calcium content
Month
28
Presentation outline
1. PhénoFinlait : general information and advances for
dairy sector
2. Normandy cows’ milk: observations from
PhénoFinlait
 Milk yield and milk composition
 Focus on fine milk composition



Fatty acids, proteins
Genetic parameters
Influence of physiological, nutritional and seasonal factors
3. Calcium content in milk : first results
4. MIR spectrometry, method to phenotype new traits
www.idele.fr
Thanks to MIR spectrometry,
milk analysis become an
indicator of herd management
Pregnancy, acidosis,
ketosis, mastitis, etc…
MIR spectrum
modify milk
reflects milk
composition
composition
Pregnancy, metabolic
diseases, methane
Milk composition
emissions…
MIR Spectrum
Milk composition gives information
on physiological status of the cows
MIR spectrum is also a general indicator of physiological status
www.idele.fr
Numerous perspectives
for use
Reproduction:
Pregnancy diagnosis
Nutrition:
Energy balance
Enteric methane emissions
Health:
Metabolic diseases (ketosis,
acidosis)
Udder health
OptiMIR Program
www.idele.fr
Technology :
Cheese-making capacity
Cheese yield
From’MIR
program
Conclusion: MIR
spectrometry to promote
Normande cows milk
quality
Fatty acids for butter production
Spreadability index (C16:0/C18:1)
Caseins for cheese production
κ et αs2 caseins
Calcium content : high and stable over
the lactation
© S. Fraisse/Cniel
www.idele.fr
Thank you for
your attention !
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