Effects of different methods of non-lethal tissue sampling on butterflies

Abstract

1. We investigated the effects of two methods of non-lethal tissue sampling on post-release flight behaviour (short-term response) and survival (longterm response) of two butterflies, Pieris rapae and Coenonympha tullia, within the same natural habitat. We applied three treatments: control (no tissue removal), wing clipping, and leg removal. Our study is the first to directly compare the effects of these common sampling methods.2. We monitored the flight behaviour of the butterflies by following individuals immediately after their release. In 99 behaviour trials of P. rapae and 101 of C. tullia we found no significant differences in proportion of time spent flying or displacement per unit time among treatment groups in either species.3. We used standard mark-recapture techniques continuously throughout the flight season to compare the survival of individuals. We marked a total of 687 P. rapae and 490 C. tullia butterflies. We found no significant differences in survival among treatments in either species. 4. We detected differences between the sexes in survival in P. rapae and flight behaviour in C. tullia. In addition to indicating differences in ecology between the sexes, these results also suggest that our analyses were sufficiently powerful to detect a significant effect of tissue removal had such an effect existed. 5. Our work is an important addition to the accumulating evidence that these methods of non-lethal tissue sampling are generally not detrimental. These sampling techniques closely mimic conditions in the wild, as wing wear and leg losses occur naturally.

Impact of non‐lethal genetic sampling on the survival, longevity and behaviour of the Hermes copper (Lycaena hermes) butterfly

“…We demonstrated that removing a prothoracic leg from Hermes copper butterflies had no measurable impact on their survival, longevity and behaviour. This is consistent with the other studies investigating non-lethal genetic sampling for various insect species (Keyghobadi et al, 2009;Hamm et al, 2010;Koscinski et al, 2011). Importantly, Hermes copper represents the smallest butterfly species for which the impact of non-lethal genetic sampling on survival and/or behaviour has been assessed thus far (Koscinski et al, 2011).…”

“…Despite the increasing use of these non-lethal sampling techniques, rarely is the impact of the tissue sampling on survival or behaviour assessed. The few studies addressing impacts on these parameters demonstrate that wing-clips and leg removal have no noticeable effects on survival and/or behaviour, especially in butterflies (Roland et al, 2000;Hamm et al, 2010;Koscinski et al, 2011). Despite this, exceptions exist with other insect groups.…”

Conservation genetics of an endangered grassland butterfly (Oarisma poweshiek) reveals historically high gene flow despite recent and rapid range loss

“…). Surveyors were trained to non‐lethally sample tissue by gently netting adults and removing a single leg from O. poweshiek after the census surveys (following protocols for non‐lethal tissue sampling of Hamm et al ., ; Koscinski et al ., ; Saarinen, ). We extracted DNA using a Qiagen DNeasy Extraction Kit (Valencia, California) and stored DNA at −80 °C until PCR experiments.…”

Analysis of tissue dependent DNA yield for optimal sampling of micro-moths in large-scale biodiversity surveys

“…However, most genetic surveys of adult insects usually extract DNA from a leg (Lai & Pullin, 2004;Watts et al, 2007;Vila et al, 2009;Koscinski et al, 2011). Indeed, legs remain the main tissue used as source of DNA in large molecular systematic studies of Lepidoptera such as the all-Lepidoptera barcoding project (Janzen et al, 2005;Burns et al, 2007; http://www.lepbarcoding.org).…”