Transformation methods for the glochidia of the spectaclecase mussel Cumberlandia monodonta

Science Center Objects

The spectaclecase mussel, Cumberlandia monodonta, was effectively listed as federally endangered in April 2012 (   It is endemic to the Mississippi, Ohio and Missouri River basins and historically inhabited 44 streams in these basins (USFWS 2014). Currently, the species is known to inhabit 20 of the historical streams, five of which are represented by one or two specimens (Butler 2002). The three remaining strongholds of the species occur in the Meramec, Gasconade and St. Croix River systems. The existence of small, geographically isolated populations has been identified in the Spectaclecase Recovery Outline (USFWS 2014) as a key threat to the species. As such, the Recovery Outline lists captive propagation  and augmentation of remaining populations as a top priority for recovery of the species. Despite numerous attempts and testing of over 60 species of fish, amphibians, crayfish and insects, a suitable host of the spectaclecase has not been identified (Knudsen and Hove 1997, Lee and Hove 1997, Hove et al. 1998, Baird 2000, Hove and Berg 2008, Hove et al. 2008, Hove et al. 2009). 

In vitro transformation of glochidia is an alternative method for production of juveniles that has been successful for over 40 species unionids (Lima et al. 2012); but has not been reported for the spectaclecase. One of the major obstacles to successful in vitro transformation of glochidia is contamination from microorganisms, particularly fungus (Owen et al. 2010, Lima et al. 2012). Current protocols for in vitro culture recommend the use of antibiotics and antimycotics to reduce contamination, but none effectively control contamination for the duration of the incubation period and some are toxic to the glochidia (Owen et al. 2010). Hydrogen peroxide has been successfully used to reduce the incidence of Saprolegniasis on fish eggs (Gaikowsld et al. 2003) and inculture of fish cell lines (Eric Leis, La Crosse Fish Health Center, La Crosse, WI, personal communication). A pilot study was conducted in November 2015 in which Lampsilis cardium glochidia were treated with 100, 500 or 1000 mg/L hydrogen peroxide for 10 minutes. The treatment effectively reduced contamination and glochidia and cells remained viable for over 1 week (personal observation). Inthe present study, pretreatment with hydrogen peroxide will be compared with controls (no hydrogen peroxide) to determine differences in glochidial viability and transformation and levels of microbial contamination in culture plates.

We will evaluate in vivo and in vitro techniques to propagate juveniles of spectaclecase mussels by inoculation of potential host species and growth in an artificial medium. Results of the study will be relevant for recovery of the spectaclecase.


  1. To generate and rank a list of potential glochidial hosts based on previous host studies and fish and amphibian survey data from the St. Croix River
  2. To determine the transformation success of spectaclecase glochidia on potential fish and amphibian species (identified in objective 1) by laboratory inoculation
  3. To evaluate in vitro culture methods, including culture media and decontamination treatments, for transformation of spectaclecase glochidia in an artificial medium.


USGS Scientist Robert Kennedy examining a mussel

USGS Scientist Robert Kennedy examining a mussel(Credit: Randy Hines, USGS UMESC. Public domain.)


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