Paternity determination in the adder (Vipera berus)—DNA fingerprinting or random amplified polymorphic DNA?

Abstract

We performed breeding experiments with adders (Vipera berus) to determine whether multiple matings may result in multiple paternity. DNA fingerprinting of mothers, their offspring, and possible fathers using a polydinucleotide probe [(TG) _n ] gave a low overall similarity between unrelated individuals (0.18±0.07; SD) and an average of 17 bands that were male-specific. In no cases were there fewer than seven paternal-specific bands present in the fingerprint of an offspring, enabling us unambiguously to identify the biological father among five males. Multiple paternity was detected in the investigated broods with offspring sired exclusively by the captive males. PCR amplification of random amplified polymorphic DNA (RAPD) using 16 decamer primers gave 76 bands and an average similarity of 0.95 (±0.01) between the males, which were collected at different, geographically well-separated localities. Although there were on average 8.3 (±1.9) bands that differ between males in pairwise comparisons, there were only 1.9 (±1.1) bands per male that are specific for a particular individual. Thus, RAPDs are adequate for paternity determination only in experiments with a low number of males, whereas DNA fingerprinting offers sufficient information to discriminate between large numbers of putative fathers.