VARIATION: DEFINITION AND

METHODS OF ESTIMATION.
ANALYSIS OF VARIANCE

Master
Produccin y Sanidad Animal
Carlos de Blas Beorlegui
c.deblas@upm.es

VARIATION

Why do we repeat experiments?

What is an experimental unit?
How much should we repeat?
Should we inform about the variation observed?
How can we express variation?
How are distributed the repetitions of individual
observations?

IS THE RESPONSE
CONSISTENT OR AT
RANDOM?

The highest the variation

more doubts about the
consistency of the response
and the highest the need of
increasing repetitions to
confirm it

SOURCES OF VARIATION
Environment: t, hr, light, NH3, noise,
animal density, microbism, cleaning

Animal: sex, breed, age/weight, indiv.

genetics, supplier, maternal influence,
health, hierarchy

Experiment: sampling, shell life,

calibration, reagents, reading mistakes,

Treatments: source/level,

VARIATION

Why do we repeat experiments?

What is an experimental unit?
How much should we repeat?
Should we inform about the variation observed?
How can we express variation?
How are distributed the repetitions of individual
observations?

IS THE RESPONSE
CONSISTENT OR AT
RANDOM?

The highest the variation

more doubts about the
consistency of the response
and the highest the need of
increasing repetitions to
confirm it

MEAN VALUES AND VARIATION

EXPERIMENT 1
Treat A
Treat B
5.15
7.15
5.10
7.10
5.00
7.00
4.90
6.90
4.85
6.85
5.00
7.00

EXPERIMENT 2
Treat A
Treat B
1
3
3
5
5
7
7
9
9
11
5.00
7.00

WEIGHT GAIN (G/D) OF GROUPS OF BROILER CHICKENS FED WITH

THREE LEVELS OF FIBRE AT TWO FEEDING FREQUENCIES
FEEDING

FIBRE

FREQUENCY

HIGH

FARM A

FARM B

39/42/37/40=40

33/35/30/38=34

HIGH

35/33/30/37=34

36/33/30/30=32

HIGH

31/25/30/32=30

30/25/27/29=28

LOW

34/31/35/35=34

32/32/34/31=32

LOW

28/25/25/30=27

24/22/23/26=24

LOW

21/20/24/19=21

18/19/17/18=18

oIS THERE EVIDENCE IN THAT F E > 6 ($ threshold)?

oTHE RESPONSE TO FIBRE LEVEL IS LINEAR?
oIS THE EFFECT OF DIET THE SAME IN EACH FARM?

IS THE RESPONSE CONSISTENT OR RANDOM?

The answers often are not obvious,
depend on variation. The statistical
response requires to determine
variation besides average values and
answer in probability terms.

treatments
P
noise

treatments

noise

VARIATION

Why do we repeat experiments?

What is an experimental unit?
How much do we repeat?
Should we inform about the variation observed?
How can we express variation?
How are distributed the repetitions of individual
observations?

WAYS TO EXPRESS VARIATION

X i X

VARIANCE S 2
GLOBAL
POPULATION

SAMPLE

n 1

2
2
X

(
X
)
/n

i
i

n 1

True variance =

2 ( X i X )2 / n
Variance estimated =

s 2 X i X / n 1

STANDARD DEVIATIN = VARIANCE = S

COEFFICIENT OF VARIATION = CV = S/X

MEAN VALUES AND VARIATION

EXPERIMENT 1
Treat A
Treat B
5.15
7.15
5.10
7.10
5.00
7.00
4.90
6.90
4.85
6.85
5.00
7.00

EXPERIMENT 2
Treat A
Treat B
1
3
3
5
5
7
7
9
9
11
5.00
7.00

S2 = 0.016

S2 = 10.0

VARIATION

Why do we repeat experiments?

What is an experimental unit?
How much do we repeat?
Should we inform about the variation observed?
How can we express variation?
How are distributed the repetitions of
individual observations?

 1
Y
e
2

xi x 2

68 %
85 %
95 %

-4

-3

-1.5
-1
-2

99 %

+1.5
+1
+2

+3

+4

THE NORMAL DISTRIBUTION

Completely defined from mean and
Mean = Mode = Median
Simetric

Smooth shape
Kurtosis = 0

Z DISTRIBUTION: EXAMPLES OF USE

Dairy cows population in Spain:
Mean = 8.000 kg/cow, S = 2.000 kg
Proportion of cows > 10.000 kg?

STANDARDIZED NORMAL DISTRIBUTION

Any normal distribution can be transformed to an
standardized normal distribution of mean = 0 and
standard deviation = 1:

xi x
Z
S
The Z distribution is tabulated and allows to
calculate the probability of how far is a given value
from the centre of the distribution.
SAS-1v

Z DISTRIBUTION: EXAMPLES OF USE

Dairy cows population in Spain:
Mean = 8.000 kg/cow, S = 2.000 kg
Proportion of cows > 10.000 kg?

Z = (10.000-8.000)/2000 = 1
Table Z: P (1) =0,8413
A 15.87% OF COWS PRODUCE > 10.000 KG

SAS ORDERS
Data valor z;
Z=probit (1-/2);run;
Proc print;
Run;
Data valor z;
Z=probit (0.975);run;
Proc print;
Run;
output z = 1.96

Z DISTRIBUTION: EXAMPLES OF USE

TABLAS FEDNA quality control
Tolerance: samples for whom the analytical
values are outside the 85% of the population
P = 0.925; Z = ~1.5

Tolerance = 1.5

C.1 PROXIMAL: % Sobre Producto Tal Cual.

ANLISIS

NOMINAL

RYE GRAIN

HUMEDAD

TOLERANCIA

PERIODICIDAD

ENSAYOS

Mx 12

Cada Lote

RD 2257/1994 n6

PROTENA BRUTA

8,9

1,5

Cada Lote

RD 2257/1994 n3 o
NF V 18-120 Dumas

EXTRACTO
ETREO

1,5

0,5

Nuevo Producto
Proveedor

RD 609/1999 n 4

FIBRA BRUTA

2,3

0,5

Nuevo Producto
Proveedor

RD 2257/1994 n7 o
Fibersac

ALMIDN

53,9

3,5

Cada Lote

Orden 16.02.00

CENIZAS

Mx 3

Nuevo Producto
Proveedor

RD 2257/1994 n12

CORNEZUELO*

Mx 0,1

Nuevo Producto
Proveedor

Test colorimtrico

PESO ESPECFICO
(kg/Hl)

Mn 70

Cada lote

Peso Especfico

*De obligado cumplimiento (orden 11.10.88)

*Si
Deelobligado
cumplimiento
(ordense
11.10.88)
resto de
especificaciones
cumple, no se tendr en cuenta este criterio
Si el resto de especificaciones se cumplen, no se tendr en cuanta este criterio

http://www.fundacionfedna.org/normas-fedna-control-calidad

 Proportion of animals weaned below a given

weight.
Proportion of animals reaching slaughter age

below or beyond a given weight.

Proportion of high producing farms (ud of
control = average/farm vs. production/cow).
Outliers detection (ex: mean 2.5 S)

Objective: Determine in a population

submitted to treatments, which part
of the variation is due to treatments
and which part is random (natural
variation among experimental units)

100

50

MODEL: Yij = mean + Ti + eij

0

A
20
30
46
28
31 (-10)

B
29
38
42
35
36 (-5)

C
90
40
42
52
56 (+15)

mean= 41

1.53

3.15

3.85

8.18

5.86

1.61

3.96

3.68

5.64

5.46

3.75

3.59

5.70

7.36

5.69

2.89

1.89

5.62

5.33

6.49

3.26

1.45

5.79

8.82

7.81

2.76

1.56

5.33

5.26

9.03

2.63

2.60

5.00

6.76

6.72

X 4.74

TOTAL variation = SST = (Xij X)2 =

[(1.53 4.74)2 + (1.53 4.74)2 + ... + (1.53 4.74)2 =
139.4
Variation due to TREATMENTS= SSt =
rj (X.j X)2 = 6 [(2.63 4.74)2 + ...+ (6.72 4.74)2] =
102.7
NATURAL variation among exp units (ERROR) = SSE =
(Xij X.j)2 = [(1.53 - 2.63 )2 +... + (3.15 2.60)2 + ...
+ (9.03 - 6.72 )2] = 36.7

TOTAL

SS

df

MS

139.4

29(n-1)

4.81

TREATM

102.7

4(k-1)

25.7

ERROR

36.7

25

1.47

17.4***
(df 4,25)

 Used for VARIANCE comparison

CI (S12 / S22) 1?= f(S12 / S22, n1, n2, sigfn):
F critic: tables
GLOBAL test of signification: indicates that at
least one of the treatmts increased appreciably
the total variation.
If the no. of treatments is high it can induce to
error.
The variances within treatments must be
homogeneous and the mean distributions be
normal.
SAS-1v

SAS ORDERS

Data valor f;
F=finv(1- , gl1, gl2);run;
Proc print;
Run;
Data valor f;
F=finv(0.95, 10, 2);run;
Proc print;
Run;
Output: Fcrit=19.3959

PROC GLM;
CLASS TRAT;
MODEL INSULIN = TRAT;

MEANS TRAT;
RUN;

Procedimiento GLM
Informacin de nivel de clase
Clase
Niveles
Valores
trat
5
a b c d e
Nmero de observaciones ledas
Nmero de observaciones usadas

30
30

Variable dependiente: Insulina

Fuente

DF

Suma de
cuadrados

Cuadrado de
la media

F-Valor

Pr > F

Modelo

102.7011000

25.6752750

17.43

<.0001

Error

25

36.8329667

1.4733187

Total correcto

29

139.5340667

R-cuadrado
0.736029

Fuente
trat

Fuente
trat

Coef Var
25.58967

DF
4

DF
4

Raiz MSE
1.213803

Insulina Media
4.743333

Tipo I SS
102.7011000

Cuadrado de
la media
25.6752750

F-Valor
17.43

Pr > F
<.0001

Tipo III SS
102.7011000

Cuadrado de
la media
25.6752750

F-Valor
17.43

Pr > F
<.0001

RECOMMENDED BOOKS
Statistical methods in Agriculture and experimental
biology (3rd ed-2003)
Mead, R., Curnow, R.N. & Hasted, A.M.
Ed. Chapman&Hall/CRC

Experimental Design and Analysis in Animal Sciences

Morris TR (1999) Ed. CABI publ.

The Design of Animal Experiments

Festing M.F.W. et al. (2010)
Laboratory Animals Handbook no 14
Ed. Laboratory Animals Ltd