PARENT SESSION
Tuesday, August 8, 5:00-6:30 pm
Poster Session 8 - Aquatic ecology
Exhibit Hall, Ballroom Level, Cook Convention Center

Who's your daddy: the use of micro-satellite analysis to determine paternity in freshwater mussels.

Smith, Allison^*,1, Loutsch, Jeannette ¹, Monroe, Emy ², Christian, Alan ¹, ¹ Arkansas State University, Jonesboro, AR² Miami University, Oxford, OH

ABSTRACT- Numbers of individuals and species of freshwater mussels have been documented as declining. While the mechanisms of release and uptake of sperm in mussels has been documented, the paternal contribution a male has on offspring of a single female is unknown. The objective of this study is to determine whether a female mussel is fertilized by one or many males. This study uses a three step process which involves: 1) establish the spatial patterns of individual male and female distributions in a stream reach, 2) non-destructively sample male and female mantle tissue and harvest glochidia from females, and 3) use micro-satellite loci to determine paternal contribution to broods based on potential males in the population. Average glochidia mass was determined to be 1.352 g (±0.22 SE). DNA was successfully extracted from glochidia using a modified Quick Extract Epicentre solution protocol with DNA quantities averaging 6.22ng/l (±0.60 SE). Successful amplification of adult DNA has occurred with 6 out of 15 previously published micro-satellite primer sets, while successful amplification of glochidia occurred for 3 out of 3 primers using an automatic fragment analyzer. Preliminary fragment analysis of gravid females and their glochidia alleles verified that mothers and their glochidia share alleles, thus proving these methods to be successful. To the authors’ knowledge, these data represent a technological breakthrough in the extraction and amplification of glochidial DNA. The results of micro-satellite paternity analysis will provide a better understanding of reproductive events and will be useful for management and conservation planning.

Key words: freshwater mussel, micro-satelite analysis, micro-satelite analysis