Investigations on saprolegniosis. II. Study on the process of penetrations showing evidence of a proteolytic enzyme and histopathological aspects [Translation from: Riv.ital.Piscicolt.Ittiopatol. 11(4), 109-117, 1976]

The study of enzymatic activity is of great importance in the immunology of fungi. Indeed, knowledge of biological activity of antigenic structures is important for the elucidation of host-parasite relations as well as in the search for a taxonomic factor permitting differential diagnoses. The authors used Saprolegnia cultures to analyse soluble antigenic fractions arising from the mycelium of cultures of 4 species of Saprolegnia, which are found most frequently in the parasitic state on fish: S. parasitica, S. ferax, S. delica, S. diclina. The authors conclude that in the study of saprolegniasis, the enzymatic approach affords new elements for the examination of the etiology of fungi as well as an element of gravity concerning the biochemical modifications necessary to the change of saprophytism to parasitism.

Molecular biomarkers and toxic consequences of impact by organic pollution in aquatic organisms

Organic contaminants are readily bioaccumulated by aquatic organisms. Exposure to and toxic effects of contaminants can be measured in terms of the biochemical responses of the organisms (i.e. molecular biomarkers). The hepatic biotransformation enzyme cytochrome P4501A (CYP1A) in vertebrates is specifically induced by organic contaminants such as aromatic hydrocarbons, PCBs and dioxins, and is involved in chemical carcinogenesis via catalysis of the covalent binding of organic contaminants to DNA (DNA-adducts). Hepatic CYP1A induction has been used extensively and successfully as a biomarker of organic contaminant exposure in fish. Fewer but equally encouraging studies in fish have used hepatic bulky, hydrophobic DNA-adducts as biomarkers of organic contaminant damage. Much less is known of the situation in marine invertebrates, but a CYPlA-like enzyme with limited inducibility and some potential for biomarker application is indicated. Stimulation of reactive oxygen species (ROS) production is another potential mechanism of organic contaminant-mediated DNA and other damage in aquatic organisms. A combination of antioxidant (enzymes, scavengers) and pro-oxidant (oxidised DNA bases, lipid peroxidation) measurements may have potential as a biomarker of organic contaminant exposure (particularly those chemicals which do not induce CYP1A) and/or oxidative stress, but more studies are required. Both CYP1A- and ROS-mediated toxicity are indicated to result in higher order deleterious effects, including cancer and other aspects of animal fitness.

Development, validation, and utilization of a competitive enzyme-linked immunosorbent assay for the detection of antibodies against Brucella species in marine mammals

A competitive enzyme-linked immunosorbent assay (cELISA) was developed by using a whole-cell antigen from a marine Brucella sp. isolated from a harbor seal (Phoca vitulina). The assay was designed to screen sera from multiple marine mammal species for the presence of antibodies against marine-origin Brucella. Based on comparisons with culture-confirmed cases, specificity and sensitivity for cetacean samples tested were 73% and 100%, respectively. For pinniped samples, specificity and sensitivity values were 77% and 67%, respectively. Hawaiian monk seal (Monachus schauinslandi; n = 28) and bottlenose dolphin (Tursiops truncatus; n = 48) serum samples were tested, and the results were compared with several other assays designed to detect Brucella abortus antibodies. The comparison testing revealed the marine-origin cELISA to be more sensitive than the B. abortus tests by the detection of additional positive serum samples. The newly developed cELISA is an effective serologic method for detection of the presence of antibodies against marine-origin Brucella sp. in marine mammals.

Prevalence of brevetoxins in prey fish of bottlenose dolphins in Sarasota Bay, Florida

Blooms of the brevetoxin-producing dinoflagellate Karenia brevis have been linked to high mortality of bottlenose dolphins Tursiops truncatus on Florida’s Gulf of Mexico coast. A clear understanding of trophic transfer of brevetoxin from its algal source up the food web to top predators is needed to assess exposure of affected dolphin populations. Prey fish constitute a means of accumulating and transferring brevetoxins and are potential vectors of brevetoxin to dolphins frequently exposed to K. brevis blooms. Here we report results of brevetoxin analyses of the primary fish species consumed by long-term resident bottlenose dolphins inhabiting Sarasota Bay, Florida. Fish collected during K. brevis blooms in 2003 to 2006 were analyzed by competitive enzyme-linked immunosorbent assay (ELISA) and had brevetoxin concentrations ranging from 4 to 10844 ng PbTx-3 eq g–1 tissue. Receptor binding assay (RBA) and liquid chromatography–mass spectrometry (LC-MS) analysis confirmed toxicity and the presence of parent brevetoxins and known metabolites. Fish collected in the absence of K. brevis blooms tested positive for brevetoxin by ELISA and RBA, with concentrations up to 1500 ng PbTx-3 eq g–1 tissue. These findings implicate prey fish exposed to K. brevis blooms as brevetoxin vectors for their dolphin predators and provide a critical analysis of persistent brevetoxin loads in the food web of dolphins repeatedly exposed to Florida red tides.

Binding of radio-iodinated gonadotropin to the ovary of exotic carp (Cyprinus carpio Lin.)

Heterologous murrel gonadotropic hormone (m-GtH) binds to common carp oocyte plasma membrane and enhances steroid secretion. With increasing concentration of radio-labelled hormone the receptor binding is also found to increase linearly up to a certain concentration and then decrease. The [¹²⁵I] murrel GtH binding characteristics to a preparation of common carp ovarian plasma membrane shows saturability with high affinity. Scat chard plot analysis gave dissociation constant (Kd) of 0.81 X 10(super -9) M and maximum binding capacity (MBC) of 22.05 f mole/mg protein.

Sulfide binding characteristics of blood serum in Calyptogena pacifica and Vesicomya gigas

The sulfide binding characteristics of blood serum were studied in vitro in two deep-sea vesicomyid clams, Calyptogena pacifica and Vesicomya gigas. Both the C. pacifica and the V. gigas serum concentrated sulfide at least an order of magnitude above ambient levels. V. gigas accumulated sulfide faster than C. pacifica, reaching saturation at 5000 M after an hour. C. pacifica bound sulfide at half the rate of V. gigas, reaching saturation in about two hours at a substantially higher concentration of sulfide. The observed distribution of the animals near cold seeps in the Monterey Submarine Canyon can be explained by their different sulfide binding abilities. The hypothesis that cold seeps are actually much more unstable sources of sulfide than previously assumed is explored.

Studies on phenolase enzyme in prawns. II. The inhibitory action on the enzyme by certain chemical agents

The effect of certain chemical agents on dopa oxidation by phenolases has been examined. Sulphur containing amino carboxylic acids are inhibitory agents for dopa oxidation. Tyrosine, a substrate for the enzyme also acts as an inhibitor for dopa oxidation by the enzyme. The possible mode of action has been discussed. The function of diethyl dithiocarbamate in suppressing the display of enzyme activity has been detailed and its behaviour has been compared to the other chemical agents studied.