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Virginia Sea Grant funded research

Reducing the mortality of horseshoe crabs in the biomedical industry’s bleeding process

Jim Berkson and Stephen Smith

Our objective is to establish a relationship between horseshoe crab size and the quantity of blood withdrawn in order to identify limits that can be applied in the biomedical industry's bleeding process to reduce crab mortalities. We will use cell culture techniques to grow amoebocytes for preparation of Limulus Amoebocyte Lysate (LAL), potentially minimizing the number of crabs that would need to be bled each year.

Horseshoe crabs housed at the Virginia-Maryland Regional College of Veterinary Medicine will be bled at assigned levels and monitored for mortalities. Relationships will be identified between bleeding level and mortality rate. We will also test for seasonality effects, then expand our study to look at a range of horseshoe crab sizes. Working with the biomedical industry, we will examiine ways to change current bleeding protocols, to reduce overall mortalities, using the results of our study. Using previously published techniques for isolating and in vitro culturing horseshoe crab amoebocytes, we will attempt to reproduce those results in our laboratory in media modified to resemble that of the horseshoe crab. Cultures will be evaluated for amoebocyte viability, internal vacuolization, cellular expansion and LAL production.

Horseshoe crabs are ecologically, medically, and economically essential components of Atlantic coast's ecosystem, but are in decline. The biomedical industry's production of LAL provides an annual social welfare benefit of $150 million to the economy. With strong support and partnership by the nation's largest producer of LAL, our study will provide ways to reduce the mortality and hopefully the need for horseshoe crabs by the biomedical industry.