Gilbert B. Pauley

The immune response in immunized and naturally infected rainbow trout (Salmo gairdneri) to Diplostomum spathaceum as detected by enzyme-linked immunosorbent assay (ELISA).

The enzyme-linked immunosorbent assay (ELISA) procedure was modified and adapted for detection of circulating antibodies in rainbow trout (Salmo gairdneri) against metacercariae of the digenean trematode Diplostomum spathaceum, the causative agent for diplostomiasis. Rainbow trout (Salmo gairdneri) were injected with sonicated metacercariae representing 10, 40, and 100 metacercariae per fish. Three weeks after immunization the average titers for trout injected with 10, 40, and 100 metacercariae were 874, 841, and 525, and by six weeks the titers had fallen to 299, 349, and 203, respectively. Nine weeks after initial immunization, two remaining fish initially immunized with 100 metacercariae per fish were injected with a booster of 50 sonicated metacercariae per fish. Four weeks later the average titer was 1204. Serum samples from naturally infected wild fish tested for the presence of circulating antibodies against Diplostomum spathaceum showed 25 of 27 with positive titers.

A preliminary physicochemical characterization of an agglutinin found in the hemolymph of the crayfish Procambarus clarkii

Abstract An agglutinin found in the serum of the crayfish Procambarus clarkii , capable of agglutinating marine bacteria and vertebrate chicken and rabbit red blood cells (RBC), was studied to determine some of its physical and chemical properties. Tests to analyze its nature were (1) p H extreme and heat stability, (2) freezing and thawing, (3) dialysis against 0.01 m Tris buffer, 0.15 M NaCl, (4) chloroform, toluene, xylene, trichloroacetic acid (TCA), ether, and phenol extraction, (5) urea, trypsin, and pronase incubation. Agglutinin activity was normal between p H 6.4 and 10.4; it was inactivated at 70°C in 30 min and by extraction with phenol and 10% TCA. Tests with RBC, three marine bacteria, and the terrestrial pathogenic bacterium Serratia marcescens showed little cross absorption activity (exception between chicken RBC and the marine bacterium 628). Agglutinin molecular weight greater than 150,000 was estimated, using Sephadex G-200 column chromatography. These results indicate that a large molecular weight material, probably containing protein, was responsible for agglutination. Crayfish serum and hemolymph were tested against two marine bacteria (628 and Fr) and one terrestrial bacterium ( S. marcescens ) for the presence of bactericidins. None of the bacteria were killed by either the serum or the hemolymph.

Experimental wound repair in the black abalone, Haliotis cracherodii☆

Histological sections of incisions made in the mantle and foot of the black abalone, Haliotis cracherodii, revealed a number of differences in wound healing of the two tissues. Initial muscular closure of the wound was more pronounced in the mantle as was invagination of the cut edges. Within 8 hr post-incision, leukocytes lined the wound surface, initiating granulation tissue formation. A layer of squamous epithelium covered this reparative tissue in the wound cavity within 16 days. Leukocytes and collagen deposition filled the foot wound cavity, pushing the new epithelium upward until it was even with the normal epithelial surface. Mucous cell secretion indicated that the new epithelium was functional 2 months after incision. Granulation tissue did not fill up the mantle wound cavity, so that the epithelial surface was still depressed 75 days post-incision. Granulation tissue formation in the foot was far more elaborate than that of the mantle during all phases of wound healing. In both structures epithelial differentation and vascularization of the granulation tissue occurred within 2 months after incision. Muscle development in the healing wound appeared to result from new muscle formation within the granulation tissue rather than by gradual infiltration of muscle fibers from the surrounding tissue. The healed tissue in both structures appeared to be functional.

Cellular defense reactions to particulate materials in the California sea hare, Aplysia californica

Abstract Cellular mechanisms of defense in a marine gastropod, the California sea hare Aplysia californica , were studied histologically following injections of either carmine or India ink. Sea hares were capable of mobilizing an efficient cellular defense against either of these foreign bodies injected intramuscularly into the foot. Carmine was gathered into large nodules and walled off, while ink was sequestered in situ within individual phagocytes. However, injections of India ink made directly into the body cavity resulted in death to these mollusks. In addition to the circulating phagocytes observed in most invertebrates, A. californica has primitive lymphoid tissue at the base of the gill.