Mirela Petrić

Insight into the Short-Finned Squid Illex coindetii (Cephalopoda: Ommastrephidae) Feeding Ecology: Is There a Link Between Helminth Parasites and Food Composition?

Abstract Squids are especially frequent as paratenic hosts of helminth parasites, particularly to those that have elasmobranchs and mammals as final hosts. Among those parasite species, anisakid nematode larvae and cestode plerocercoids are most effectively transferred through the trophic chain by oegopsid squids. A total of 439 short-finned squids, Illex coindetii (245 males, 190 females and 4 unsexed) were sampled in the central part of the eastern Adriatic Sea in order to assess their helminth component community and parasite dynamics with respect to host sex, maturity, seasonality, and feeding behavior. Two larval helminths were isolated, i.e., larvae of Anisakis pegreffii, characterized by molecular tools at the species level, and plerocercoids of Phyllobothrium sp., with prevalences of 30.5% and 2.3%, respectively. Highly significant seasonal variation in diet consumption, congruent with seasonal variation in anisakid intensity, was observed, underlining the tight role of squid prey in the trophic transmission of parasite. Likewise, the highest helminth prevalence and intensity of infection was recorded in autumn, when the fish prey, mostly Maurolicus muelleri, comprised the greatest proportion of diet. This helped to assign the Adriatic broadtail shortfin squid not as a first, but as a second, paratenic host for the anisakid, unlike as suggested previously. The presence of larval A. pegreffii confirms its previously reported zoogeographical distribution in the Mediterranean and Adriatic Seas. The presence of 2 helminths in I. coindetii describes the feeding patterns of the squid, as well as clearly defined and coevolved predator–prey relationships.

Scarcity of parasite assemblages in the Adriatic-reared European sea bass (Dicentrarchus labrax) and sea bream (Sparus aurata)

The shaping forces of parasite community structure still is the main subject in the ecological parasitology whilst community predictability and repeatability showed that hardly a generally applicable role is ever going to be assessed. Defining and describing parasite communities can be very useful from the epizootiological point, in order to help in the assessment of the medical and economical impact of certain parasitosis, moreover when hosts are economically valuable species. Since parasite assemblages in reared European sea bass (Dicentrarchus labrax) and sea bream (Sparus aurata) in Adriatic cage systems can play an important role in the economic feasibility of the rearing process, we evaluated their character through assessing diversity indices, nestedness of parasite communities and their differences in respect to season and composition, as well as fish growth. We observed colonization of a new monogenean species (Furnestinia echeneis) and general impoverishment of parasites populations over time in the Adriatic-reared fish parasite assemblages. Parasite assemblages differed significantly between seasons for both fish species, while species richness, evenness, diversity indices and nestedness of parasitic communities in the sea bream showed to be significantly higher compared to those in the sea bass. Such characteristics define parasite communities of both Adriatic-reared fish as species poor although structured and ordered assemblages.

Reaction of the mussel Mytilus galloprovincialis (Bivalvia) to Eugymnanthea inquilina (Cnidaria) and Urastoma cyprinae (Turbellaria) concurrent infestation

In total 480 individuals of Mytilus galloprovincialis were sampled monthly from October 2009 to September 2010, at the shellfish farm in the Mali Ston Bay, south Adriatic Sea (Croatia) in order to assess the extent of pathology imposed by two parasites, Eugymnanthea inquilina (Cnidaria) and Urastoma cyprinae (Turbellaria). Although a deteriorating impact on host reproduction or condition index was lacking, we evidenced ultrastructural and functional alteration in host cells at the attachment site. Ultrastructural changes included hemocytic encapsulation of the turbellarian and cell desquamation in medusoid infestation. Caspase positive reaction inferred by immunohistochemistry (IHC) was triggered in cases of turbellarian infestation, in contrast with hydroids, suggesting that the former exhibits more complex host-parasite interaction, reflected in the persistent attempts of the parasite to survive bivalve reaction. We have evidenced that both organisms trigger specific host reaction that although not costly in terms of host reproductive cycle or growth, results in mild tissue destruction and hemocyte activation. A lower degree of tissue reaction was observed in cases of hydroid infestation, compared to turbellarian.

Growth and reproduction of Munida rutllanti (Decapoda: Anomura: Galatheidae) and impact of parasitism by Pleurocrypta sp. (Isopoda: Bopyridae) in the Adriatic Sea

Relative growth and reproductive biology of Munida rutllanti were investigated for the first time in the Adriatic Sea. A total of 93 8 individuals were analysed. Sex-ratio differed statistically from the expected 1:1. Six individuals exhibited secondary sex characteristics of both males and females. Carapace length of males ranged from 10.5 to 21.5 mm, while in females it ranged from 11.2 to 19.4 mm. Sexual dimorphism was observed in seven morphometric characters. Ovigerous females were first observed in summer and later in autumn with higher frequency. Realized fecundity showed wide variation (273 - 3250 eggs) and positive correlation between the brood and female size. An epicaridean isopod Pleurocrypta sp., parasitizing the gill chamber of both sexes was isolated and its effect on the oogenesis was studied by histological examination of host reproductive system. Growth inhibition, modification of secondary sex characteristics, as well as retardation of oogenesis and failure of oviposition, are repercussions of bopyrid infestation. Given the small prevalence ( 7 .8 5 %) of the bopyrid in the M. rutllanti Adriatic population, we assume that for the moment there is no severe effect on the host population dynamic.

The effect of artificial feed on blood biochemistry profile and liver histology of wild saddled bream, Oblada melanura (Sparidae)

Floating fish farms attract a great number of wild fish species, changing their behaviour and physiology. The saddled bream, Oblada melanura, sampled from populations aggregated around the Adriatic fish farm and from natural/control populations, were analysed for differences in eleven blood biochemistry parameters and liver histomorphology. The levels of lactate dehydrogenase (LDH), alkaline phosphatase (ALKP) and urea (URE) in cage-associated saddled bream (428.00±SD 321.56 U/L, 86.13±SD 39.87 U/L and 0.05±SD 0.16 mmol/L, respectively) were significantly lower than those observed in the control specimens (1047.06±SD 505.56 U/L, 125.75±SD 34.70 U/L and 1.99±SD 0.73 mmol/L, respectively). In contrast to that, concentrations of aspartate aminotransferase (AST) in cage-associated fish (87.63±SD 132.34 U/L) were higher than values noted for the control population (6.55±SD 5.90 U/L). URE and AST presented the main variables contributing to the discrimination between two analysed populations. One-way ANOSIM based on the blood parameters showed significant difference between saddled bream that fed around cages and those from the remote waters (R=0.697; P < 0.01). Hepatocytes of cage-associated fish contained large cytoplasmatic clear spaces indicating excessive accumulation of fat in the hepatocyte cytoplasm. All observed differences can be attributed to contrasting feeding behaviour of sampled populations but basic nutritional differences between them should be quantified in the future. Moreover, further research is necessary to detect their impact on the health status of the fish.

Multixenobiotic Resistance Mechanism in Gills of Reared vs. Wild Mediterranean Mussel Mytilus galloprovincialis (Lamarck, 1819)

Mussel (Mytilus galloprovincialis (Lamarck, 1819)) is directly exposed to sea water contamination that elicits significant physiological and cellular response, although its extent mounted in aquaculture-reared in comparison to wild bivalve populations is scarcely known. Therefore, we have compared contamination biomarkers in mussels from reared (Marina farm) and wild, anthropogenically affected site (Vranjic Bay). While predictably, the levels of metals (Cu, Cd, Pb, Zn, Fe, and Hg) in whole bivalve tissues determined by atomic absorption spectrophotometry resulted in significantly higher concentrations in wild mussels, accompanied by elevated number of apoptotic cells in gills, the activity of multixenobiotic resistance defense mechanism (MXR), measured as the accumulation rate of model substrate rhodamine B (RB) gave contrasting results. The functional RB assay evidenced a lower MXR efflux activity in the gill tissue of wild mussels, indicating two possible scenarios that will need further focus: (1) persisting sea water pollution increased cell damage of bivalve gill cells and consequently led to leakage of the RB into cytoplasm and dysfunctional MXR efflux in wild mussels; or/and (2) a mixture of different toxic compounds present in Vranjic Bay sea water induced oversaturation of MXR efflux, inducing elevated accumulation of the dye. Consequently, it seems that an efficient physiological functioning of MXR in wild mussels is strongly hampered by existence of an unknown quantity of sea water pollutants that may endanger intrinsic organismal defense system and lead toward the enhancement of toxicity.