Differentiation between larvae of Tipula paludosa and Tipula oleracea (Diptera: Tipulidae) using isoelectric focusing, and their occurrence in grassland in northern Britain

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Abstract
Summary: Thin layer agarose isoelectric focusing (IEF) was used to separate proteins from the larvae of Tipula oleracea Linnaeus and Tipula paludosa Meigen (Diptera, Nematocera). Silver staining revealed protein banding patterns which consistently distinguished these two species at any stage in their life cycles. Within the pH range 5–6 Tipula paludosa contained one major protein band and T. oleracea two bands, with the pI value of the T. paludosa protein being slightly higher than those of T. oleracea. The IEF method is particularly useful for the larval stages of these species because they are often visually inseparable. Leather jackets obtained during a survey of Northern Ireland and north‐eastern and western Scotland were tested to determine the distribution and frequency of these species occurring in grassland. In each area surveyed T. paludosa predominated with only one field in Northern Ireland and north‐eastern Scotland found to contain T. oleracea. In western Scotland T. oleracea occurred in five fields each year of the survey, and was locally abundant on the Isle of Bute in the 1990/91 survey making up 15% of the total larvae tested.