Calcium-dependent proteinases of some invertebrates and fishes

In comparative-evolutionary aspect, the experimental data are considered about activity, biochemical properties, and peculiarities of structural organization of proteins of the calpain family in some invertebrates and fishes. Peculiarities of calpain-like proteins of invertebrates--the predecessors of calpains of higher animals are revealed. By the example of the studied taxons, there is traced complication of the structural organization and mechanisms of control of the calpain activities, which reflects stages of molecular evolution of the protein family.     

Toxicity of rotenone to some species of coarse fish and invertebrates

The toxicity of rotenone was determined for seven species of fish and three species of invertebrates. The results show that there is a marked variation in sensitivity of different species to rotenone. The survival times of roach Rutilus rutilus (L.) were reduced by a rise in temperature and also by a reduction in water hardness. Temperature affected the rate at which rotenone is degraded; 3 days after being prepared a solution containing 2 mg/1 of 5 % rotenone was non-toxic to roach over a 7-day period at 20°C, but it was still toxic for at least 11 days at 11.5°C. The data are discussed in relation to the use of rotenone in fisheries management.     

Aspartic proteinases in fishes and aquatic invertebrates

1. The literature on molecular properties and physiological role of aspartic proteinases in fishes and aquatic invertebrates has been reviewed. 2. Pepsins have not been detected in invertebrates, and apparently cathepsin D, as well as other cathepsins, act both as digestive and lysosomal enzymes in many of these animals. The molecular properties of invertebrate cathepsin D correspond with cathepsin D in fishes and mammalians. 3. Fishes with a true stomach have pepsinogen secretion. Fish pepsins have higher pH optimum and are less stable in strong acid conditions than mammalian pepsins. They are very efficient at low temperatures, but less thermostable than mammalian pepsins. 4. Many fishes have two significantly different pepsins: Pepsin I and Pepsin II, which digest haemoglobin at a maximal rate in the pH ranges 3-4 and 2-3 respectively. Usually the pI of Pepsin I is in the range 6.5-7, whereas pI of Pepsin II is about 4. 5. Fish Pepsin I and cathepsin D have very similar molecular properties, and a hypothesis proposing that cathepsin D is the ancestor enzyme of aspartic proteinases in higher animals is presented.     

Ubiquinone analyses in fish tissues and in some marine invertebrates.

1. Ubiquinone contents were determined in species of marine invertebrates, and in heart, red and white muscle and liver of three species of fish. 2. Three different methods of determination were compared, based on spectrophotometry, reduction and a reaction with the dimethoxy groups of ubiquinone. 3. Using ubiquinone homologues 6-10 prepared from beef heart and commercially available microorganisms (SCP) as standards, ubiquinone 10 was found in all samples. In addition were found minor amounts of Q-9 in samples of saithe heart and red muscle. 4. Less than 10 mg/kg wet wt of ubiquinone was found in the samples of marine invertebrates and in white muscle and liver of the fish samples, with one exception: 40 mg/kg in a sample of mackerel liver. 5. Higher contents of ubiquinone were found in fish heart and red muscle tissues, ranging from 24 to 116 mg/kg wet wt. The ubiquinone contents were comparable in the two tissues. 6. A test on cellular fragments of red muscle tissue of saithe showed that the ubiquinone was concentrated in the mitochondria fraction.     

Aspartic proteinases in Antarctic fish

The present review surveys several recent studies of the aspartic proteinases from Antarctic Notothenioidei, a dominating fish group that has developed a number of adjustments at the molecular level to maintain metabolic function at low temperatures. Given the unique peculiarities of the Antarctic environment, studying the features of Antarctic aspartic proteinases could provide new insights into the role of these proteins in fish physiology. We describe here: (1) the biochemical properties of a cathepsin D purified from the liver of the hemoglobinless icefish Chionodraco hamatus; (2) the biochemical characterization of Trematomus bernacchii pepsins variants A1 and A2 obtained by heterologous expression in bacteria; and (3) the identification of two closely related, novel aspartic proteinases from the liver of the two Antarctic fish species mentioned above. Overall, the results show that Notothenioidei aspartic proteinases display a number of characteristics that are remarkably different from those of mammalian aspartic proteinases, including high turnover number or high catalytic efficiency. We have named the newly identified aspartic proteinases "Nothepsins" and classified them relative to aspartic proteinases from other species.     

Vibrios of some deep-water invertebrates

Abstract The gut microbiota of 7 species of deep-water (300–400 m) invertebrates from the Gulf of Mexico was examined. High populations of Vibrio spp. were observed in crustaceans (ranging from 10⁵ to 7 × 10⁶ cells/g gut content) while relatively low populations of Vibrio spp. were found in annelids, the water column, and sediment. Although saprophytic Vibrio species were isolated, Vibrio fluvialis and Vibrio hollisae , potential human pathogens, were isolated from the crustaceans, Pleoticus robustus, Nematocarcinas sp., Plesionika sp., and Munida sp., and Vibrio vulnificus was isolated from Nereis sp. These observations confirm the finding of Ohwada et al. [18] that the gut of deep-water invertebrates has a bacterial flora abundant in Vibrio spp. These results also suggest that some marine invertebrates may serve as reservoirs for certain potential pathogenic Vibrio species.     

Toxicity of synthetic detergents to fish and aquatic invertebrates

Synthetic detergents are reported to be acutely toxic to fish in concentrations between 0.4 and 40 mg/1. Factors affecting toxicity include the molecular structure of the detergent, water hardness, temperature and dissolved oxygen concentration; the age and species of the test fish, and acclimation to low concentrations of detergent. Some of these factors appear to be of only limited importance. Gill damage is the most obvious acute toxic effect; the immediate cause of death may be asphyxiation, but detergents may also be toxic internally. Lethal effects not related to gill damage have not been investigated. Sublethal effects include retardation of growth, alteration of feeding behaviour and inhibition of chemoreceptor organs. Low levels of detergents may also increase the uptake of other pollutants. Invertebrates, especially in their juvenile stages, are extremely sensitive to detergents: concentrations below 0.1 mg/1 interfere with growth and development in some species. The interactions between detergents and proteins, and their influence on membrane permeability may be the basis of the biological action of detergents. Detergents in natural waters are usually partially degraded, and a maximum permissible concentration of 0.5 mg/1 would probably be harmless under most conditions.     

Studies on the proteolytic enzymes of invertebrates constituting fish food

Studies were carried out on the proteolytic activity of trypsin and pepsin-like enzymes of invertebrates consituting fish food. Relations between proteolytic activity of enzymes, pH, and temperature were established.     

The interaction between some serine proteinases and horse leucocyte inhibitor

Horse blood leucocyte cytosol exhibits a broad inhibitory activity against serine proteinases. The purified inhibitor was exposed to investigated enzymes (trypsin, chymotrypsin, elastases and serine proteinase from S. aureus) for variable time and the products were analyzed by gradient polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate. The molar ratio I:E, association rate constants k on and inhibition constants Ki for the enzymes and inhibitor were determined. The examined elastases form stable, stoichiometric complexes with the inhibitor (Ki less than 10(-10) M), and do not undergo proteolytic degradation during 30 min incubation at 20 degrees C even at the 2-fold molar excess of the proteinases. The reactions with elastases are extremely rapid (k on greater than 10(7) M-1 s-1) and are completed within one second whereas similar reactions with chymotrypsin and trypsin are much slower (k on = 3 X 10(5) M-1 s-5 and 5 X 10(2) M-1 s-1, respectively). Serine proteinase from S. aureus neither react nor inactivates the investigated inhibitor. The complexes of the inhibitor with trypsin and chymotrypsin are digested even at a molar ratio I:E = 2:1. All these observations point out that the inhibitor from horse leucocyte cytosol is a specific and effective inhibitor of elastases.     

Helminth infections of some invertebrates of the Georgia Bight

Dominant macroinvertebrates of the Georgia Bight were studied to evaluate the incidence of helminth infection and pathologies produced. Twenty-one stations along seven transects were sampled in February, May, August, and November 1977. Sicyonia brevirostris (rock shrimp) were heavily infected (83%) with cestode cysticercoids in the hepatopancreas and nerve tracts. Loligo pealeii (common squid) were lightly infected with cestodes (6%) throughout the year and with trematodes (<5%) during most of the year, but in the fall 33% were parasitized with trematode metacercarial. This study reemphasizes the high frequency of infection of marine invertebrates over large areas, the considerable pathology produced as well as the remarkable variability in infection patterns in local areas and with season.