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Background. Proteolytic enzymes may serve multiple functions: they may inhibit the host′s blood clotting, protect the parasite from the host ′s immune response, facilitate parasite′s migration within a tissue by decomposing the tissue barrier, enhance the hatching and moulting of larvae, and play an important role in their feeding. Learning their identity leads to a better understanding of a host-parasite system. The objective of this study was to check, using biochemical methods, if, in addition to proteases, ES products and extracts of 3rd and 4th larval stages of Anisakis simplex (Rudolphi, 1809) contain other hydrolases. Materials and Methods. Stage-3 larvae (L 3 ) of A.simplex were removed from Baltic herring, Clupea harengus membras Linnaeus, 1761 caught in the Baltic Sea. Stage-4 larvae (L 4) were obtained from an L 3 stage culture kept in Eagle ′s medium. The solutions containing ES products were collected and dialysed at 4°C against distilled water for 24 h. Larval extracts were obtained by homogenising the larvae in a physiological saline (0.9 % NaCl) solution in a glass homogeniser. The homogenate was centrifuged for 10 min at 3000 G. The supernatant was used in enzyme activity assays. Enzymatic activity of ES products and homogenates of L 3 and L 4 larvae of A. simplex was determined with the API ZYM test. Results. The excretion-secretion product of L 3 and L 4 larvae of A. simplex revealed activities of 10-and 11 hydrolases, respectively. Activity of esterase, esterase lipase, valine arylamidase, and N-acetyl-ß-glucosaminidase in the L 4 larvae extracts was higher than the activity of a corresponding enzyme assayed in the L 3 extracts. Only in the case of acid phosphatase, its activity in L 3 ES products was twice that of the activity found in ES products of L 4 larvae. Enzymes such as trypsin, chymotrypsin, and ß-glucosidase were not detected in extracts from L 3 larvae. Conclusion. Activities of most hydrolases in the L 4 extracts were higher than the activities of corresponding enzymes assayed in the L 3 extracts. The high activity of these enzymes found in L 4 larval extracts could be related to a different feeding mechanism of stage-4 larvae.
In the years 1994-1996, a total of 592 specimens of herring Clupea harengus membras L., caught in southern Baltic, were examined. The investigations revealed four species of trematodes: Hemiurus lühei, H. raabei, H. levinseni, Brachyphallus crenatus, two species of nematodes: Hysterothylacium auctum (syn. H. aduncum), and Anisakis simplex and three acanthocephalans: Echinorhynchus gadi, Metechinorhynchus salmonis and Pomphorhynchus sp. The finding of H. levinseni constitutes the first record of this species in Polish waters. The infection parameters of herring (prevalence, intensity and abundance) were compared in relation to different capture areas and seasons of the year. Possible affects of the feeding ground, fish body length and the spawning group of herring on the infection levels were also examined. No relation was observed between the place and the season of catch and the infection level of herring. The fish representing different feeding grounds (Baltic, Danish Straits) and sequential length classes showed variable parameters of infection.
The concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and dioxin-like polychlorinated biphenyls (DL-PCB) were determined in samples of Baltic herring (Clupea harengus membras) and sprat (Sprattus sprattus balticus) in 2006 from commercial catches in Estonian coastal waters, Baltic Sea. The dioxin content of the fish sampled in 2006 did not exceed the European Union’s maximum permissible level for PCDD/Fs (4.0 pg WHO -TEQ/g fresh weight) and for the sum of PCDD/Fs and DL-PCBs (8.0 pg WHO -TEQ/g fresh weight). PCDD/Fs and the sum of PCDD/Fs and DL-PCBs content in herring were 2.12 and 3.84 pg WHO-TEQ/g of fresh weight respectively; the corresponding figures for sprat were 1.94 and 3.82 pg WHO-TEQ/g of fresh weight. Comparable with our earlier data on the content of dioxins in three to four year old herring and two to three year old sprat, these data show that two servings of fish per week are not at all harmful to the health of the Estonian people; indeed, the opposite is more likely to be the case.
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