I also made contact subsequent to arriving back in the USA in Jan 2002 with William S. Marshall, Professor, Biology Department, St. Francis Xavier University, Antigonish NS  Canada, who is a world expert in voltage clamping of Fundulus opercular membrane.  He could be an important colleague to help in studying the lobster bronchial membrane, which might be an important cuticular surface that is attacked in shell disease.  Being one of the thinnest membranes of the lobster the opercular membrane might be the first cuticular surface to be penetrated and might underlie a hitherto unknown mechanism of death that could be part of the shell disease syndrome.  I will add this membrane to the list of structures to be examined in shell diseased lobsters.
 

The Online Database of lobster work has been added to from data and samples taken on Leg IV of AL02-03 Spring 2002 Bottom Survey by Joseph G. Kunkel as a volunteer scientist.  436 lobsters were measured and 108 special samples of serum were taken with ovarian samples taken on all light orange to orange or green serum with annotations on serum samples plus random control ovaries taken from clear serum females.  This material was quick frozen and will be analyzed this summer by a recruited undergraduate.
 

The timing of lobster egging-out was focused on in sampling done aboard the Albatross IV in the Gulf of Maine lobster population.  Each lobster was observed through its ventral intersegmental membranes, Fig 1,  to see if the orange color of the hemolymph could be observed non-invasively.  Light orange to orange hemolymph as observed through this cuticle triggered a further sampling to confirm the serum color.  The serum was taken with a 3 ml syringe through the dorsal carapace right of center intersegmental membrane with the abdomen.  The hemolymph was immediately filtered through a syringe tip filter and the color of that hemolymph noted.  If the serum was yellow to orange or (in 1 case green) the ovaries of the lobster were taken and frozen.  The color of the ovaries was noted.  Orange to light green ovaries were only found in females with light orange hemolymph.  Orange to dark orange hemolymph was an indicator of advanced ovary development (dark green and firm with well developed eggs).  The one green hemolymph female had flaccid dark green ovaries which will be examined in the lab for the status of its eggs.  The control females with clear serum all had dark green firm ovaries whose egg status must also be determined in the lab.
 

My current hypothesis about serum and ovary color is somewhat changed from previous hypotheses.  A female who is beginning to deposit vitellogenin from the blood into developing eggs has a light orange hemolymph that is often seen as clear hemolymph through the cuticle.  Its ovaries may be orange to yellowish-green to green but the ovary is small in diameter (0.5 cm at most.  The hemolymph is orange because production of Vg by the hepatopancreas is ahead of uptake into the ovaries and thus the orange Vg can be seen in the serum.  When ovaries are maximally growing the hemolymph is in generally clear because the ovaries are efficiently clearing Vg from the hemolymph.  When the eggs are chorionating the uptake of Vg stops and some Vg piles up in the serum and the serum may become dark orange.  The ovaries of dark orange serum females are always large in diameter with visible eggs within the ovarian epithelium.  The one female with green hemolymph had flaccid large ovaries which I suspect were involuting.  There were no signs of ovulated eggs in this female.  The green hemolymph females are most often found in pounds and in caged captive females and I continue to believe these are not healthy reproducing females.  Analysis of the hemolymph and ovarian samples this summer will allow a better assessment of this current hypothesis.  Each frozen ovary will be examined to measure the diameter of the developing oocytes and the status of enzymatic processing of its Vg peptides.
 

A collaboration with Dr. Diane Cowan to extend our interest in the normal serum patters of lobsters to shell disease is developing.  I talked with a DVM, Stephen Ellis, employed by Maine Department of Marine Resources working on the aquacultured salmon diseases.  He was sampling fish caught on the Albatross while in the northern Gulf of Maine for the viral disease organisms that is currently attacking Maine salmon pens.  He told me of a shell disease outbreak in Maine waters which occurred several years ago and discussed various scenarios that might have contributed to the outbreak.  While that outbreak has passed, the current outbreak spreading from Long Island Sound is possibly related and some greater collaboration with the Maine lobster experience should be explored.  The collaboration with Diane Cowan becomes more important and sampling in both the Massachusetts area as well as the central Gulf of Maine lobster areas is warranted.
 

Personnel

Dr. Diane Cowan. President of The Lobster Conservency collaborated in providing lobsters that could be bled at regular intervals.

Jeff Xu applied his skills at database management improving an online database of Lobster serum collection information.

Dennis Carrol, a UMass undergraduate has worked in the lab as a lab technician, mainly helping to keep the lab glassware and working spaces orderly.  He has also learned to take images of specimens using our image acquisition system.  His current project will be to photograph the ovary samples and catalog the lobster serum specimens brought back from the AL02-03 leg IV cruise.