ALBERT

Notes: SUMMARY. 1. The reliability of the simple frequency, Janetschek, Cassie and Dyar's law methods for determining or corroborating instars of mayflies and stoneflies was evaluated using data from published studies, a population of Baetisca rogersi and populations simulated through use of random numbers and generated normal distributions.2. The Janetschek and Cassie methods are variations of the simple frequency method that offer no significant advantage. Modes of the Cassie method, thought to represent instars, are much more difficult or impossible to detect than are the corresponding peaks of the other two methods.3. Overlap in size between adjacent instars can lead to false instar peaks or modes in frequency plots. The potential for overlap in mayflies and stoneflies is greatly increased, compared to other insects, because of their large number of instars and known developmental variability. The normal distribution simulations demonstrated that instar size variability as low as 5–7.5% COV (coefficient of variability) may lead to false instar peaks when the number of instars is in the typical range. These simulations also indicated that even simple frequency plots with distinct peaks may result in inaccurate instar determinations.4. The number of size classes used in an analysis was correlated with the number of peaks or modes revealed. The number of peaks greater than zero in the Janetschek plots for the Baetisca rogersi population varied from 5 to 53 as the number of size classes was varied from 20 to 188. Similarly for the random number simulations. the number of peaks varied from 6 to 41 as the number of size classes varied from 22 to 127.5. Dyar's law semi-logarithmic plots do not corroborate instars determined through frequency methods, because the uniform spacing of‘instar’data points is the direct result of the uniform spacing of peaks in frequency plots of most data sources (including random numbers), whether or not peaks actually indicate instars. Also Dyar's law plots will‘corroborate’different numbers of instars depending on the peak selection criteria used. The potential for corroborating instars through supplemental rearing and best-fit analysis is discussed.6. The future of mayfly—stonefly instar determination lies in the increased and more rigorous application of the rearing and Palmen body (mayflies only) methods.