DESIGN _3

LATIN SQUARE DESIGN

Latin Square Design

More than one blocking variables

Consider 2 blocking variables to be used simultaneously to reduce

experimental error associated with each blocking variable
Example
Age
Age-A:20-29
Age-B:30-39
Age-C:40-49

Lifestyle
LS-1:Sedentary
LS-2: Quite active
LS-3: Very active

Example
Observer
Observer1
Observer2
Observer3

Day
Monday
Tuesday
Wednesday

Example 1: Persons as experimental units

A study to assess three experimental diets (treatments)

containing varying fat content on lipid level in blood.
Male subjects who are within 20% of their ideal body
weight are chosen. However their ages vary, and so are
their lifestyles. Need to block age (say 3 groups) and
lifestyle (say 3 groups).
Data on reduction in lipid level are taken after a certain
period of time on the diet.

Suppose you want to use RCBD

Block

LS-1
T2 T1 T3
Age-A
LS-1
T2 T1 T3
Age-B

LS-1
T3 T2 T1
Age-C
LS-2
T1 T3 T2
Age-A

LS-3
T2 T3 T1
Age-C

RCBD has 9 blocks and 27

experimental units.
The disadvantage of using
RCBD to block two variables
simultaneously is that it
requires too many
experimental units.

Is there a way to reduce the

number of experimental unit
to cut cost?

Lets consider using Latin Square design (an extreme

form of Incomplete Block design)
Latin square design:
one treatment per block
LS-1

LS-2

Features of a Latin square

design

LS-3

Age-A

T2

T1

T3

Age-B

T1

T3

T2

Age-C

T3

T2

T1

There are r treatments

There are two blocking
variables, each with r classes
Each row and each column
contain all treatments; each
class of each blocking variable
constitutes a replication.

Example 2: Field experiment -plots as experimental

units
The layout is straight forward because blocking is associated
with experimental setting. The blocking variables are obvious.

A
Forest

Dirt road

Not
dusty

dusty

Yet another setup of Latin squares

Fish in cages. The cages arranged in rows along the

river. The positions in cages represent columns.
A B C D
B A D C
C D B A

D C A B

ANOVA
Model Yijk = + i + j + k + ijk
where i=j=k = 1,., r and i = j = k = 0
Source

df

SS

Row

r-1

(iY2i../r) - (Y2/r2)

SSR/SSE

Column

r-1

(jY2.j./r) - (Y2/r2)

SSC/SSE

Treatment

r-1

(kY2..k/r) - (Y2/r2)

SSTR/SSE

Error

(r-1)(r-2)

Total

r2-1

by diff.

ijkYijk2 - (Y2/r2)

Note: error df for 3x3 squares = 2. For small latin square experiment,
additional replications (ie. repeating the experiment) are required.

Size of squares

The most common square in the range 5x5 to 8x8

Larger square above 12 x12 rarely used

Small squares provide few df for estimation of experimental error

e.g. 3 x 3 squares
1 square

2 replicating squares

squares

s-1

=1

row

row(square)

r-1(s)

=4

column

column(square)

r-1(s)

=4

treatment

treatment

r-1

=2

error

error

total

total

=6
sr2-1

=17

Worked example
Column
Row

C1

C2

C3

C4

C5

Total

R1

B
257

E
230

A
279

C
287

D
202

1255

R2

D
245

A
283

E
245

B
280

C
260

1313

R3

E
182

B
252

C
280

D
246

A
250

1210

R4

A
203

C
204

D
227

E
193

B
259

1086

R5

C
231

D
271

B
266

A
334

E
338

1440

Total

1118

1240

1297

1340

1309

6304

Computation of SS

A
1349

Summary of treatment

B
1314

C
1262

D
1191

E
1188

Sum of Squares

CF = 63042/25 = 1,589,617
Total SS
= (2572 + .+3382) 1,589,617 = 36,571
Row SS
= (12552 + .+ 14402)/5 1,589,617 = 13,601
Column SS
= (11182 + ..+ 13092)/5 1,589,617 = 6,146
Treatment SS
= (13492 + ..+ 11882)/5 1,589,617 = 4,156
Error SS
= 12,668