Experimental Design in Agriculture CROP 590, Winter, 2016 Why conduct experiments?... To explore new technologies, new crops, and new areas of production To develop a basic understanding of the factors that control production To develop new technologies that are superior to existing technologies To study the effect of changes in the factors of production and to identify optimal levels To demonstrate new knowledge to growers and get feedback from end-users about the acceptability of new technologies What is a designed experiment? Treatments are imposed (manipulated) by investigator using standard protocols May infer that the response was due to the treatments Potential pitfalls As we artificially manipulate nature, results may not generalize to real life situations As we increase the spatial and temporal scale of experiments (to make them more realistic), it becomes more difficult to adhere to principles of good experimental design What is an observational study? Treatments are defined on the basis of existing groups or circumstances Uses – Early stages of study – developing hypotheses – Scale of study is too large to artificially apply treatments (e.g. ecosystems) – Application of treatments of interest is not ethical May determine associations between treatments and responses, but cannot assume that there is a cause and effect relationship between them Testing predictions in new settings may further support our model, but inference will never be as strong as for a designed (manipulative) experiment Some Types of Field Experiments (Oriented toward Applied Research) Agronomy Trials – – – – Fertilizer studies Time, rate and density of planting Tillage studies Factors are often interactive so it is good to include combinations of multiple levels of two or more factors – Plot size is larger due to machinery and border effects Integrated Pest Management – Weeds, diseases, insects, nematodes, slugs – Complex interactions betweens pests and host plants – Mobility and short generation time of pests often create challenges in measuring treatment response Types of Field Experiments (Continued) Plant Breeding Trials – Often include a large number of treatments (genotypes) – Initial assessments may be subjective or qualitative using small plots – Replicated yield trials with check varieties including a long term check to measure progress Pasture Experiments – Initially you can use clipping to simulate grazing – Ultimately, response measured by grazing animals so plots must be large – The pasture, not the animal, is the experimental unit Types of Field Experiments (Continued) Experiments with Perennial Crops – Same crop on same plot for two or more years – Effects of treatments may accumulate – Treatments cannot be randomly assigned each year so it is not possible to use years as a replication – Large plots will permit the introduction of new treatments Intercropping Experiments – Two or more crops are grown together for a significant part of the growing season to increase total yield and/or yield stability – Treatments must include crops by themselves as well as several intercrop combinations – Several ratios and planting configurations are used so number of treatments may be large – Must be conducted for several years to assess stability of system Types of Field Experiments (Continued) Rotation Experiments – Determine effects of cropping sequence on target crop, pest or pathogen, or environmental quality – Treatments are applied over multiple cropping seasons or years, but impact is determined in the final season Farming Systems Research – – – – – – To move new agricultural technologies to the farm A number of farms in the target area are identified Often two large plots are laid out - old versus new Should be located close enough for side by side comparisons May include “best bet” combinations of several new technologies Recent emphasis on farmer participation in both development and assessment of new technologies The Scientific Method Formulation of an H ypothesis P lanning an experiment to objectively test the hypothesis Careful observation and collection of D ata from the experiment I nterpretation of the experimental results Steps in Experimentation H Definition of the problem Statement of objectives P Selection of treatments Selection of experimental material Selection of experimental design Selection of the unit for observation and the number of replications Control of the effects of the adjacent units on each other Consideration of data to be collected Outlining statistical analysis and summarization of results D Conducting the experiment I Analyzing data and interpreting results Preparation of a complete, readable, and correct report The Well-Planned Experiment Simplicity – don’t attempt to do too much – write out the objectives, listed in order of priority Degree of precision – appropriate design – sufficient replication Absence of systematic error Range of validity of conclusions – well-defined reference population – repeat the experiment in time and space – a factorial set of treatments also increases the range Calculation of degree of uncertainty Hypothesis Testing H0: = ɵ HA: ɵ or H0: 1= 2 HA: 1 2 /2 /2 critical value If the observed (i.e., calculated) test statistic is greater than the critical value, reject H0 If the observed test statistic is less than the critical value, fail to reject H0 Hypothesis Testing The concept of a rejection region (e.g. = 0.05) is not favored by some statisticians It may be more informative to: – Report the p-value for the observed test statistic – Report confidence intervals for treatment means Hypothesis testing It is necessary to define a rejection region to determine the power of a test Decision Accept H0 Reject H0 Reality H0 is true 1 = 2 Correct HA is true 1 2 1- Type II error Power Type I error Power of the test H0 HA 1– p /2 1– /2 Power is greater when – differences among treaments are large – alpha is large – standard errors are small Review - Corrected Sum of Squares Definition formula n SS Y Yi Y i1 2 Computational formula – common in older textbooks uncorrected sum of squares correction factor Yi n i1 2 SS Y Yi n i1 n 2 t tests – one sample To test the hypothesis that the mean of a single population is equal to some value: Y 0 t sY Compare to critical t for n-1 df for a given (0.05 in this graph) standard error of the mean s2 where s Y n df = df = n-1 df = 6 df = 3 t tests – two samples, equal variance To compare the mean of two populations with equal variances and equal sample sizes: Y1 Y 2 t s Y1 Y2 standard error of the difference where s Y1 Y2 2 P 2s n df = 2(n-1) The pooled s2 (sP2 ) should be the average of the variance estimates of the two samples t tests – unequal sample size To compare the mean of two populations with equal variances and unequal sample sizes: Y1 Y 2 t sY1 Y2 where s Y1 Y2 1 1 s df = (n1-1) + (n2-1) n1 n2 2 P The pooled s2 (sP2 ) should be a weighted average of the variance estimates from the two samples 2 2 𝑛 − 1 𝑠 + 𝑛 − 1 𝑠 𝑆𝑆1 + 𝑆𝑆2 1 2 1 2 𝟐 𝒔𝑷 = = 𝑛1 − 1 + 𝑛2 − 1 𝑛1 − 1 + 𝑛2 − 1 Paired t test When observations are paired, it may be beneficial to use a paired t test – for example, feeding rations given to animals from the same litter t2 = F in a Completely Randomized Design (CRD) when there are only two treatment levels Paired t2 = F in a RBD (Randomized Complete Block Design) with two treatment levels Measures of variation Sample variance Standard deviation SSY s n1 2 s Coefficient of Variation 2 s CV% x 100 Y – Expresses the standard deviation as a proportion of the mean – Varies from 0 to 100 (not on original scale) – Used to compare precision of data from similar types of experiments Confidence Intervals 100(1-)% confidence interval for a mean (𝐿𝑌 ) Y t ,df 2 s n If we repeatedly obtained samples of the same size from a population, and constructed confidence intervals, we would expect 95% of those intervals to contain the true mean. 100(1-)% confidence interval for a difference between two means (𝐿𝑌1−𝑌2 ) Y Y t 1 2 ,df 2s2 n Measures of Variation s (standard deviation) Y s2 n CV (coefficient of variation) s2 * 100 Y t ,df se (standard error of a mean) L (Confidence Interval for a mean) 2 s n sY1Y2 t ,df 2 2 (standard error of a difference between means) 2s2 n Y t ,df t ,df s2 n 2 LSD (Least Significant Difference between means) L(Confidence Interval for a difference between means) t ,df 2s2 n Y Y t 1 2 ,df 2s2 n