Characterization of a new platelet aggregating factor from crotoxin Crotalus durissus cascavella venom

In this article we investigated the platelet aggregating activity of whole crotoxin and its subunits isolated from Crotalus durissus cascavella venom. During the purification protocols of the venom, using HPLC molecular exclusion, we detected the presence of two different serine protease activities in the gyroxin fraction, and another in the crotoxin fraction, which induced strong and irreversible platelet aggregation, in addition to blood coagulation. From crotoxin, we isolated PLA(2), crotapotin (both fractions corresponding approximately 85% of whole crotoxin) and another minor fraction (F20) that exhibited serine protease activity. After a new fractionation on reverse phase HPLC chromatography, we obtained three other fractions named as F201, F202 and F203. F202 was obtained with high degree of molecular homogeneity with molecular mass of approximately 28 kDa and a high content of acidic amino residues, such as aspartic acid and glutamic acid. Other important amino acids were histidine, cysteine and lysine. This protein exhibited a high specificity for BApNA, a Michaelis-Menten behavior with Vmax estimated in 5.64 mu M/min and a Km value of 0.58 mM for this substrate. In this work, we investigated the ability of F202 to degrade fibrinogen and observed alpha and beta chain cleavage. Enzymatic as well as the platelet aggregation activities were strongly inhibited when incubated with TLCK and PMSF, specific inhibitors of serine protease. Also, F202 induced platelet aggregation in washed and platelet-rich plasma, and in both cases, TLCK inhibited its activity. The N-terminal amino acid sequence of F202 presented a high amino acid sequence homology with other thrombin-like proteins, but it was significantly different from gyroxin. These results showed that crotoxin is a highly heterogeneous protein composed of PLA(2), thrombin-like and other fractions that might explain the diversity of physiological and pharmacological activities of this protein.

Ocorrência e tratamento de monogenoides em alevinos de pampo (Trachinotus carolinus) cultivados experimentalmente na região norte do estado de São Paulo

This work had as objective tests therapeutic treatments seeking the monogenea eradication in fingerlings Florida Pompano (Trachinotus carolinus) servants in cage net in the area of Ubatuba, State of São Paulo. The fingerlings presented an accentuated weigh loss and strong discoloration and through observation of scraped of gills under light microscope were identified a high amount of monogenea. Three treatments were tested in form of baths: T1 (fresh water for 5 minutes); T2 (formalin: 1: 1.000 for 20 minutes) and T3: (formalin: 1:4.000 for 30 minutes). The treatments were appraised through scraped of gills, mounted among you laminate and laminulas and observed to the light microscope. It was observed that in the tested conditions all of the treatments were efficient in the elimination of the monogenea without presenting lethality to the fish. The treatment is recommended T I (take a bath in fresh water for 5 minutes) for the facility in the application and for the absence of use of chemical products.

Eugenol como anestésico para jundiá (Rhamdia voulezi) em diferentes pesos

The study aimed to find the better concentration of eugenol for anesthesia of silver catfish (Rhamdia voulezi) with different weights. Were used 240 catfish distributed in randomized blocks in factorial scheme (5x4) total 20 treatments, in others words, five different weights: 32,5; 75; 150; 300 e 450g and four eugenol concentrations: 50, 75, 100 e 125 mg.l(-1). For each treatment were used 12 fish randomly chosen and exposed individually for each concentration. After of anesthesia the fish were transferred for net-cage with 0,7m(3), being fed and observed by 96 hours for monitoring of mortality. The eugenol was efficient for anesthesia in silver catfish all concentrations and weights and after of 96 hours no mortality have been verified. At these experimental conditions the best concentration of eugenol for anesthetic inducing and recuperation of silver catfish with weight varying from 32,5 and 450 g is 50 mg.l(-1).

Nesting biology of the fungus growing ants Mycetarotes Emery (Attini, Formicidae)

Fungus-growing ants of the genus Mycetarotes are among the least studied in the tribe Attini. This report documents nest architecture and worker population numbers for 19 nests of M. parallelus and 5 nests of M. acutus, including the first such report for M. acutus. This new information is integrated with the scant biological information reported on Mycetarotes to date. The resulting picture of Mycetarotes life history, as well as the relative ease with which large numbers of nests can be collected and observed in the field, suggest that Mycetarotes (particularly M. parallelus) is an ideal model system for the study of coevolution of lower-attine ants and their cultivated fungi.