Latin Square Designs (15.

4)

Lecture Objective
Introduce basic experimental designs that
account for two orthogonal sources of
extraneous variation.
Terminology
Square design
Orthogonal blocks
Randomizations
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 Examples

A researcher wishes to perform a yield experiment under field

conditions, but she/he knows or suspects that there are two
fertility trends running perpendicular to each other across the
study plots.
An animal scientists wishes to study weight gain in piglets but
knows that both litter membership and initial weights
significantly affect the response.
In a greenhouse, researchers know that there is variation in
response due to both light differences across the building and
temperature differences along the building.
An agricultural engineer wishing to test the wear of different
makes of tractor tire, knows that the trial and the location of the
tire on the (four wheel drive, equal tire size) tractor will
significantly affect wear.
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 Latin Square Design

A class of experimental designs that allow for two sources

of blocking.
Can be constructed for any number of treatments, but there
is a cost. If there are t treatments, then t2 experimental
units will be required.
If one of the blocking factors is left out of the design, we are
left with a design that could have been obtained as a
randomized block design.
Analysis of a Latin square is very similar to that of a RBD,
only one more source of variation in the model.
Two restrictions on randomization.

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 Cold Protection of Strawberries

Three different irrigation methods (treatment

levels) are used on strawberries:
1. drip,
2. overhead sprinkler,
3. no irrigation.
We wish to determine which of these is most
effective in protecting strawberries from
extreme cold.
All strawberries grown through plastic mulch.
Measure weight of frozen fruit (lower values
indicate more protection).
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 high Nitrogen Level low

Field

Moisture
none drip over

Layout none over none

drip drip over

Moisture and Soil Nitrogen CANAL
are two sources of
extraneous variation that
we wish to simultaneously
control for. high Nitrogen Level low

none drip over

Moisture

Which design will drip over none

best allow us to
account for both soil over none drip
moisture and
CANAL
nitrogen gradients?
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 Advantages and Disadvantages

Advantages:
Allows for control of two extraneous sources of
variation.
Analysis is quite simple.

Disadvantages:
Requires t2 experimental units to study t treatments.
Best suited for t in range: 5 t 10.
The effect of each treatment on the response must be
approximately the same across rows and columns.
Implementation problems.
Missing data causes major analysis problems.
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 Constructing a Latin Square Design for t Treatments

Treatments designated by first t capital letters in the alphabet

(A,B,C, etc.)
Number the levels of blocking factor 1 (call it Rows) as R1,
R2, Rt.
Number the levels of blocking factor 2 (call it Columns) as
C1, C2, Ct.
Assign the treatment letters in alphabetic order, beginning
with A, to the t units in the first row.
For the second row, start with the letter B and assign
treatment letters to the t-th letter then follow with A.
For rows 3 through t, simply shift the treatment letters up one
at a time, placing the shifted letter in the last unit of the row.

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 Basic Square

C1 C2 C3 C4
R1 A B C D
R2 B C D A
R3 C D A B
R4 D A B C

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 Randomization

C1 C2 C3 C4
Get a random
ordering of the R1 B C D A
rows.
R2 A B C D
1234 R3 D A B C
replaced by R4 C D A B
2143

Reorder the rows according to randomization.

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 Randomization

C1 C2 C3 C4
Get a random
ordering of the R1 A C D B
columns. R2 D B C A
1234 R3 C A B D
replaced by R4 B D A C
4231

Reorder the columns according to randomization.

Two Blocking Factors = Two Randomizations
= Two Constraints on Randomization
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 Latin Square Linear Model: A Three-Way AOV

yij ( k ) i j k ij ( k ) , (i, j , k 1, , t )

t = number of treatments, rows and columns.

yij(k) = observation on the unit in the ith row, jth column given the kth
treatment. The indicator k is in parenthesis to remind us that specifying
i and j effectively determines the treatment k.
= the general mean common to all experimental units.
i = the effect of level i of the row blocking factor. Usually assumed
N(0,2), a random effect.
j = the effect of level j of the column blocking factor. Usually
assumed N(0,2), a random effect.
k = the effect of level k of treatment factor, a fixed effect.
ij(k) = component of random variation associated with observation ij(k).
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Usually assumed N(0,2).
Latin Square Analysis of Variance

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 Sums of Squares
t
TSS (y
i , j 1
ij ( k ) y ) 2

t
SSRow t ( yi.(.) y ) 2

i 1
t
SSCol t ( y. j (.) y ) 2

j 1
t
SSTrt t ( yij (.) y ) 2

k 1
SSE TSS SSRow SSCol SSTrt
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Experimental Error

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 Latin Square Mean Squares and F Statistics

SSRow
MSRow =
(t - 1) MSTrt
Ftrt ~ F( t 1),( t 1)( t 2 )
SSCol MSE
MSCol = MSRow
(t - 1) Frow ~ F( t 1),( t 1)( t 2 )
MSE
SSTrt MSCol
MSTrt = Fcol ~ F( t 1),( t 1( t 2 )
(t - 1) MSE
SSE
MSE =
((t - 1)(t - 2)
We reject the null hypothesis of no main effect if the value of
the F-statistic is greater than the 100(1-)th percentile of the F
distribution with degrees of freedom specified above.
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 Latin Square Example

The strawberry irrigation cold protection study data are given

below. The effectiveness of the three irrigation methods was
measured by the weight of the frozen fruit, with lower weights
representing more effective protection. The study question is
Which irrigation method provided the most protection?
Row Column Treatment Weight
1 1 Drip 51
1 2 Over 119
1 3 None 60
2 1 None 98
2 2 Drip 43
2 3 Over 31
3 1 Over 99
3 2 None 87
3 3 Drip 49

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Data strawb;
input row column irrig $ weight @@;
datalines;
1 1 drip 51 1 2 over 119 1 3 none 60
2 1 none 98 2 2 drip 43 2 3 over 31 Latin Square in
3 1 over 99 3 2 none 87 3 3 drip 49
; run;
SAS
proc glm;
class row column irrig;
model weight = row column irrig;
title 'Strawberry Irrigation Latin Square Exp'; run;

Sum of
Source DF Squares Mean Square F Value Pr > F
Model 6 5840.000000 973.333333 1.20 0.5205
Error 2 1621.555556 810.777778
Corrected Total 8 7461.555556

R-Square Coeff Var Root MSE weight Mean

0.782679 40.23037 28.47416 70.77778

Source DF Type I SS Mean Square F Value Pr > F

row 2 817.555556 408.777778 0.50 0.6648
column 2 2616.222222 1308.111111 1.61 0.3826
irrig 2 2406.222222 1203.111111 1.48 0.4026

Source DF Type III SS Mean Square F Value Pr > F

row 2 817.555556 408.777778 0.50 0.6648
column 2 2616.222222 1308.111111 1.61 0.3826
irrig 2 2406.222222 1203.111111 Latin1.48
Square -17 0.4026
Input Data
Analyze > General Linear Model > Univariate Latin Square in SPSS

Note: You must use a custom model and

only ask for main effects.

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SPSS Ouptut

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Latin Square in Minitab Stat > ANOVA > General Linear Model

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 MTB: ANOVA and Sums of Squares

General Linear Model: weight versus row, column, irrig

Factor Type Levels Values

row fixed 3 1, 2, 3
column fixed 3 1, 2, 3
irrig fixed 3 drip, none, over

Analysis of Variance for weight, using Adjusted SS for Tests

Source DF Seq SS Adj SS Adj MS F P

row 2 817.6 817.6 408.8 0.50 0.665
column 2 2616.2 2616.2 1308.1 1.61 0.383
irrig 2 2406.2 2406.2 1203.1 1.48 0.403
Error 2 1621.6 1621.6 810.8
Total 8 7461.6

S = 28.4742 R-Sq = 78.27% R-Sq(adj) = 13.07%

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 Latin Square with R

> straw <- read.table("Data/latin_square.txt",header=TRUE)

> straw.lm <- lm(weight ~ factor(row) + factor(column) +
factor(irrig), data=straw)
> anova(straw.lm)

Analysis of Variance Table

Response: weight
Df Sum Sq Mean Sq F value Pr(>F)
factor(row) 2 817.56 408.78 0.5042 0.6648
factor(column) 2 2616.22 1308.11 1.6134 0.3826
factor(irrig) 2 2406.22 1203.11 1.4839 0.4026
Residuals 2 1621.56 810.78

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Medical Example of a Latin Square

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 Randomized, controlled,
double-blinded, NICE!

Design extracts out

differences due to time and
patients!

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