The ANOVA Procedure

Example 25.2 Alternative Multiple Comparison Procedures

The following is a continuation of the first example in the section One-Way Layout with Means Comparisons. You are studying the effect of bacteria on the nitrogen content of red clover plants, and the analysis of variance shows a highly significant effect. The following statements create the data set and compute the analysis of variance as well as Tukey’s multiple comparisons test for pairwise differences between bacteria strains; the results are shown in Figure 25.1, Figure 25.2, and Figure 25.3

title1 'Nitrogen Content of Red Clover Plants';
data Clover;
   input Strain $ Nitrogen @@;
   datalines;
3DOK1  19.4 3DOK1  32.6 3DOK1  27.0 3DOK1  32.1 3DOK1  33.0
3DOK5  17.7 3DOK5  24.8 3DOK5  27.9 3DOK5  25.2 3DOK5  24.3
3DOK4  17.0 3DOK4  19.4 3DOK4   9.1 3DOK4  11.9 3DOK4  15.8
3DOK7  20.7 3DOK7  21.0 3DOK7  20.5 3DOK7  18.8 3DOK7  18.6
3DOK13 14.3 3DOK13 14.4 3DOK13 11.8 3DOK13 11.6 3DOK13 14.2
COMPOS 17.3 COMPOS 19.4 COMPOS 19.1 COMPOS 16.9 COMPOS 20.8
;
proc anova data=Clover;
   class Strain;
   model Nitrogen = Strain;
   means Strain / tukey;
run;

The interactivity of PROC ANOVA enables you to submit further MEANS statements without re-running the entire analysis. For example, the following command requests means of the Strain levels with Duncan’s multiple range test and the Waller-Duncan k-ratio t test.

   means Strain / duncan waller;
run;

Results of the Waller-Duncan k-ratio t test are shown in Output 25.2.1.

Output 25.2.1: Waller-Duncan K-ratio t Test

Nitrogen Content of Red Clover Plants

The ANOVA Procedure
 
Waller-Duncan K-ratio t Test for Nitrogen

Kratio 100
Error Degrees of Freedom 24
Error Mean Square 11.78867
F Value 14.37
Critical Value of t 1.91873
Minimum Significant Difference 4.1665

Means with the same letter are
not significantly different.
Waller Grouping Mean N Strain
  A 28.820 5 3DOK1
         
  B 23.980 5 3DOK5
  B      
C B 19.920 5 3DOK7
C        
C D 18.700 5 COMPOS
  D      
E D 14.640 5 3DOK4
E        
E   13.260 5 3DOK13


The Waller-Duncan k-ratio t test is a multiple range test. Unlike Tukey’s test, this test does not operate on the principle of controlling Type I error. Instead, it compares the Type I and Type II error rates based on Bayesian principles (Steel and Torrie, 1980).

The Waller Grouping column in Output 25.2.1 shows which means are significantly different. From this test, you can conclude the following:

  • The mean nitrogen content for strain 3DOK1 is higher than the means for all other strains.

  • The mean nitrogen content for strain 3DOK5 is higher than the means for COMPOS, 3DOK4, and 3DOK13.

  • The mean nitrogen content for strain 3DOK7 is higher than the means for 3DOK4 and 3DOK13.

  • The mean nitrogen content for strain COMPOS is higher than the mean for 3DOK13.

  • Differences between all other means are not significant based on this sample size.

Output 25.2.2 shows the results of Duncan’s multiple range test. Duncan’s test is a result-guided test that compares the treatment means while controlling the comparison-wise error rate. You should use this test for planned comparisons only (Steel and Torrie, 1980). The results and conclusions for this example are the same as for the Waller-Duncan k-ratio t test. This is not always the case.

Output 25.2.2: Duncan’s Multiple Range Test

Alpha 0.05
Error Degrees of Freedom 24
Error Mean Square 11.78867

Number of Means 2 3 4 5 6
Critical Range 4.482 4.707 4.852 4.954 5.031

Means with the same letter are
not significantly different.
Duncan Grouping Mean N Strain
  A 28.820 5 3DOK1
         
  B 23.980 5 3DOK5
  B      
C B 19.920 5 3DOK7
C        
C D 18.700 5 COMPOS
  D      
E D 14.640 5 3DOK4
E        
E   13.260 5 3DOK13


Tukey and Least Significant Difference (LSD) tests are requested with the following MEANS statement. The CLDIFF option requests confidence intervals for both tests.

   means Strain/ lsd tukey cldiff ;
run;

The LSD tests for this example are shown in Output 25.2.3, and they give the same results as the previous two multiple comparison tests. Again, this is not always the case.

Output 25.2.3: T Tests (LSD)

Nitrogen Content of Red Clover Plants

The ANOVA Procedure
 
t Tests (LSD) for Nitrogen

Alpha 0.05
Error Degrees of Freedom 24
Error Mean Square 11.78867
Critical Value of t 2.06390
Least Significant Difference 4.4818

Comparisons significant at the 0.05 level are indicated by ***.
Strain
Comparison
Difference
Between
Means
95% Confidence Limits  
3DOK1 - 3DOK5 4.840 0.358 9.322 ***
3DOK1 - 3DOK7 8.900 4.418 13.382 ***
3DOK1 - COMPOS 10.120 5.638 14.602 ***
3DOK1 - 3DOK4 14.180 9.698 18.662 ***
3DOK1 - 3DOK13 15.560 11.078 20.042 ***
3DOK5 - 3DOK1 -4.840 -9.322 -0.358 ***
3DOK5 - 3DOK7 4.060 -0.422 8.542  
3DOK5 - COMPOS 5.280 0.798 9.762 ***
3DOK5 - 3DOK4 9.340 4.858 13.822 ***
3DOK5 - 3DOK13 10.720 6.238 15.202 ***
3DOK7 - 3DOK1 -8.900 -13.382 -4.418 ***
3DOK7 - 3DOK5 -4.060 -8.542 0.422  
3DOK7 - COMPOS 1.220 -3.262 5.702  
3DOK7 - 3DOK4 5.280 0.798 9.762 ***
3DOK7 - 3DOK13 6.660 2.178 11.142 ***
COMPOS - 3DOK1 -10.120 -14.602 -5.638 ***
COMPOS - 3DOK5 -5.280 -9.762 -0.798 ***
COMPOS - 3DOK7 -1.220 -5.702 3.262  
COMPOS - 3DOK4 4.060 -0.422 8.542  
COMPOS - 3DOK13 5.440 0.958 9.922 ***
3DOK4 - 3DOK1 -14.180 -18.662 -9.698 ***
3DOK4 - 3DOK5 -9.340 -13.822 -4.858 ***
3DOK4 - 3DOK7 -5.280 -9.762 -0.798 ***
3DOK4 - COMPOS -4.060 -8.542 0.422  
3DOK4 - 3DOK13 1.380 -3.102 5.862  
3DOK13 - 3DOK1 -15.560 -20.042 -11.078 ***
3DOK13 - 3DOK5 -10.720 -15.202 -6.238 ***
3DOK13 - 3DOK7 -6.660 -11.142 -2.178 ***
3DOK13 - COMPOS -5.440 -9.922 -0.958 ***
3DOK13 - 3DOK4 -1.380 -5.862 3.102  


If you only perform the LSD tests when the overall model F test is significant, then this is called Fisher’s protected LSD test. Note that the LSD tests should be used for planned comparisons.

The TUKEY tests shown in Output 25.2.4 find fewer significant differences than the other three tests. This is not unexpected, as the TUKEY test controls the Type I experimentwise error rate. For a complete discussion of multiple comparison methods, see the section Multiple Comparisons in Chapter 42: The GLM Procedure.

Output 25.2.4: Tukey’s Studentized Range Test

Alpha 0.05
Error Degrees of Freedom 24
Error Mean Square 11.78867
Critical Value of Studentized Range 4.37265
Minimum Significant Difference 6.7142

Comparisons significant at the 0.05 level are indicated by ***.
Strain
Comparison
Difference
Between
Means
Simultaneous 95% Confidence
Limits
 
3DOK1 - 3DOK5 4.840 -1.874 11.554  
3DOK1 - 3DOK7 8.900 2.186 15.614 ***
3DOK1 - COMPOS 10.120 3.406 16.834 ***
3DOK1 - 3DOK4 14.180 7.466 20.894 ***
3DOK1 - 3DOK13 15.560 8.846 22.274 ***
3DOK5 - 3DOK1 -4.840 -11.554 1.874  
3DOK5 - 3DOK7 4.060 -2.654 10.774  
3DOK5 - COMPOS 5.280 -1.434 11.994  
3DOK5 - 3DOK4 9.340 2.626 16.054 ***
3DOK5 - 3DOK13 10.720 4.006 17.434 ***
3DOK7 - 3DOK1 -8.900 -15.614 -2.186 ***
3DOK7 - 3DOK5 -4.060 -10.774 2.654  
3DOK7 - COMPOS 1.220 -5.494 7.934  
3DOK7 - 3DOK4 5.280 -1.434 11.994  
3DOK7 - 3DOK13 6.660 -0.054 13.374  
COMPOS - 3DOK1 -10.120 -16.834 -3.406 ***
COMPOS - 3DOK5 -5.280 -11.994 1.434  
COMPOS - 3DOK7 -1.220 -7.934 5.494  
COMPOS - 3DOK4 4.060 -2.654 10.774  
COMPOS - 3DOK13 5.440 -1.274 12.154  
3DOK4 - 3DOK1 -14.180 -20.894 -7.466 ***
3DOK4 - 3DOK5 -9.340 -16.054 -2.626 ***
3DOK4 - 3DOK7 -5.280 -11.994 1.434  
3DOK4 - COMPOS -4.060 -10.774 2.654  
3DOK4 - 3DOK13 1.380 -5.334 8.094  
3DOK13 - 3DOK1 -15.560 -22.274 -8.846 ***
3DOK13 - 3DOK5 -10.720 -17.434 -4.006 ***
3DOK13 - 3DOK7 -6.660 -13.374 0.054  
3DOK13 - COMPOS -5.440 -12.154 1.274  
3DOK13 - 3DOK4 -1.380 -8.094 5.334