Pseudoreplication is defined. as the use of inferential statistics to test for treatment effects with data from experiments where either treatments are not replicated (though samples may be) or replicates are not statistically independent. In ANOVA terminology, it is the testing for treatment effects with an error term inappropriate to the hypothesis being considered. Scrutiny of 176 experi- mental studies published between 1960 and the present revealed that pseudoreplication occurred in 27% of them, or 48% of all such studies that applied inferential statistics. The incidence of pseudo- replication is especially high in studies of marine benthos and small mammals. The critical features of controlled experimentation are reviewed. Nondemonic intrusion is defined as the impingement of chance events on an experiment in progress. As a safeguard against both it and preexisting gradients, interspersion of treatments is argued to be an obligatory feature of good design. Especially in small experiments, adequate interspersion can sometimes be assured only by dispensing with strict random- ization procedures. Comprehension of this conflict between interspersion and randomization is aided by distinguishing pre-layout (or conventional) and layout-specifit alpha (probability of type I error). Suggestions are offered to statisticians and editors of ecological j oumals as to how ecologists' under- standing of experimental design and statistics might be improved.

/pdf/pseudoreplication-and-the-design-of-ecological-field-44o8kzdaoi.pdf

Pseudoreplication and the Design of Ecological Field Experiments

This example is provided so that non-theorists may see actual applications of the theory previously described. The Dickcissel sex ratio is employed as an indirect index of suitability. A sex ratio index was found to be correlated positively with density. This is consistent with the hypothesis that territorial behavior in the males of this species limits their density. This study provides a valid example of how the problem can be approached and offers a first step in the eventual identification of the role of territorial behavior in the habitat distribution of a common species.

/pdf/on-territorial-behavior-and-other-factors-influencing-6u6llonho3.pdf

On territorial behavior and other factors influencing habitat distribution in birds

The statistical literature on tests to compare treatments after the analysis of variance is reviewed, and the use of these tests in ecology is examined. Monte Carlo simulations on normal and lognormal data indicate that many of the tests commonly used are inappropriate or inefficient. Particular tests are recommended for unplanned multiple comparisons on the basis of controlling experimentwise type I error rate and providing maximum power. These include tests for parametric and nonparametric cases, equal and unequal sample sizes, homogeneous and heterogeneous variances, non-independent means (repeated measures or adjusted means), and comparing treatments to a control. Formulae and a worked example are provided. The problem of violations of assumptions, especially variance heterogeneity, was investigated using simulations, and particular strategies are recommended. The advantages and use of planned comparisons in ecology are discussed, and the philosophy of hypothesis testing with unplanned multiple comparisons is consid- ered in relation to confidence intervals and statistical estimation.

https://www.fc.up.pt/pessoas/amsantos/support/dayquin1989.pdf

Comparisons of Treatments After an Analysis of Variance in Ecology

Automated simultaneous determination of ammonia and total amino acids in ruminal fluid and in vitro media

In response to the need for rapid, inexpensive, high-throughput assays for antimycobacterial drug screening, a microplate-based assay which uses Alamar blue reagent for determination of growth was evaluated. MICs of 30 antimicrobial agents against Mycobacterium tuberculosis H37Rv, M. tuberculosis H37Ra, and Mycobacterium avium were determined in the microplate Alamar blue assay (MABA) with both visual and fluorometric readings and compared to MICs determined in the BACTEC 460 system. For all three mycobacterial strains, there was < or = 1 dilution difference between MABA and BACTEC median MICs in four replicate experiments for 25 to 27 of the 30 antimicrobics. Significant differences between MABA and BACTEC MICs were observed with 0, 2, and 5 of 30 antimicrobial agents against H37Rv, H37Ra, and M. avium, respectively. Overall, MICs determined either visually or fluorometrically in MABA were highly correlated with those determined in the BACTEC 460 system, and visual MABA and fluorometric MABA MICs were highly correlated. MICs of rifampin, rifabutin, minocycline, and clarithromycin were consistently lower for H37Ra compared to H37Rv in all assays but were similar for most other drugs. M. tuberculosis H37Ra may be a suitable surrogate for the more virulent H37Rv strain in primary screening of compounds for antituberculosis activity. MABA is sensitive, rapid, inexpensive, and nonradiometric and offers the potential for screening, with or without analytical instrumentation, large numbers of antimicrobial compounds against slow-growing mycobacteria.

/pdf/microplate-alamar-blue-assay-versus-bactec-460-system-for-4s09rn421d.pdf

Microplate alamar blue assay versus BACTEC 460 system for high-throughput screening of compounds against Mycobacterium tuberculosis and Mycobacterium avium.

R. G. D. Steel

Papers

Principles and procedures of statistics.