Beauvericin cytotoxicity to the invertebrate cell line SF-9

The cyclic hexadepsipeptide beauvericin, initially known as a secondary metabolite produced by the entomopathogenic fungus Beauveria bassiana and toxic to Artemia salina larvae, has been more recently recognized as an important mycotoxin synthesized by a number of Fusarium strains, which parasite maize, wheat and rice. Therefore, this mycotoxin may enter the food chain, causing yet unknown effects to the health of both domestic animals and humans. The cytotoxic effects of beauvericin on mammalian cells have been studied. We investigated the cytotoxicity of this compound in an in vitro invertebrate model, viz. the insect cell line SF-9 (immortalized pupal ovarian cells of the lepidopter Spodoptera frugiperda). Cultures of SF-9 cells in the stationary phase were exposed to beauvericin at concentrations ranging from 100 nM to 300 microM, for different periods of time (from 30' to 120 h). The effects on cell viability were assessed by the trypan blue exclusion method. After 4 h of incubation no significant decrease in cell viability was recorded in SF-9 cell cultures exposed to low concentrations of beauvericin, i.e. 100 nM and 300 nM. However, a slight decrease in viability (3.9%) was seen already in cells exposed to the mycotoxin at the 1 microM concentration. This effect became gradually more evident at higher concentrations (approximately equal to 28% at 30 microM, approximately equal to 50% at 100 microM, approximately equal to 68% at 300 microM). An even more pronounced reduction in cell viability was observed after a 24 h exposure. Under these conditions, 1 microM beauvericin caused an approx. 10% decrease in the number of viable cells, which became more significant at higher concentrations approximately equal to 23% at 3 microM, approximately equal to 47% at 10 microM, approximately equal to 65% at 30 microM, approximately equal to 90% at 100 microM, approximately equal to 99% at 300 microM). Therefore, the 50% cytotoxic concentrations (CC50) at 4 h and 24 h could be estimated as 85 microM and 10 microM, respectively. In time-course experiments, no effect of beauvericin (30 microM) on cell viability could be seen after exposure for periods of time as long as 30', 1 h and 2 h, respectively. In contrast, when SF-9 cells were exposed to the mycotoxin for longer periods of time, from 8 h to 120 h, we recorded a strong cytotoxic effect already in the low micromolar concentration range. Thus, the CC50 after both 72 h and 120 h exposure times was assessed as 2.5 microM. Higher concentrations caused a virtually 100% cell death. The data collected suggest that beauvericin exerts a substantial dose- and time-dependent cytotoxic effect on invertebrate cells, comparable to the effects described in mammalian cells.     

Selectivity Parameters and Size at First Maturity in Deep-Water Shrimps, Aristaeomorpha foliacea (Risso, 1827) and Aristeus antennatus (Risso, 1816), from the North-Western Ionian Sea (Mediterranean Sea)

Selectivity experiments were carried out during trawling targeting deep-water shrimps Aristaeomorpha foliacea (Risso, 1827) and Aristeus antennatus (Risso, 1816) (Crustacea, Decapoda, Aristeidae) in the North-Western Ionian Sea (Eastern-Central Mediterranean). Different criteria were employed to analyse maturity; however, the proportion at 50% of retained, mated and mature specimens was always used to indicate the size, expressed as Carapace Length (CL, mm), at first capture (CL_c), mating (CL_sp) and at first maturity (CL_m), respectively. In order to estimate the size at 50% maturity (CL_m) for females of both species, three criteria were adopted. In particular, CL_m was computed for the mature females not considering the presence of spermatophores, for the mature females with spermatophores and for the mature females intersected by the decreasing proportion with size of females without spermatophores. Three diamond stretched mesh codends of 40, 50 and 60 mm were tested using a cover of 20 mm. The 40-mm stretched mesh size (European Union legal size in the Mediterranean) was not selective for the sampled population of each species. The size at first capture (CL_c), calculated in both species for the two sexes combined, increased significantly with mesh size. Even for the mesh size of 60 mm, the size at first capture was still smaller than the sizes at 50% maturity, whatever the criterion adopted. Since the differences between the size at first maturity and the sizes at first capture are greater in A. foliacea than A. antennatus, the former species appears in this respect to be more vulnerable to trawling than the latter.     

Catch Composition on Red Shrimps’ (Aristaeomorpha foliacea and Aristeus antennatus) Grounds in the Eastern Ionian Sea

In the present study, the catch composition and the catch per unit effort (CPUE) by weight and numbers in red shrimps’ (Aristaeomorpha foliacea and Aristeus antennatus) grounds was examined in the southern part of the eastern Ionian Sea, in order to collect important information for the Greek waters, where no deep-water fishery exists. In the depth stratum 500–700 m, the catch of the commercial species represented a high proportion (>70%) of the total catch. Red shrimps and several other commercial species were found in important quantities. The present results suggest the possibility of developing a deep-water fishery in Greece. In such a case, attention should be paid because of the high vulnerability of A. foliacea – the main deep-water fishing resource in the area – to the fishing pressure.     

Reproduction of macrourids in the upper slope of the north-western Ionian Sea

Aspects of reproduction of macrourid fish in the north-western Ionian Sea are reported. Data were collected during 12 trawl surveys carried out between 1993 and 1995. Mature and running ripe females of Coelorhynchus coelorhynchus, Hymenocephalus italiens and Nezumia sclerorhynchus were found all year round. Mature females of Trachyrhynchus trachyrhynchus were found mainly in August.     

A meta‐analysis of isolation by distance: relic or reference standard for landscape genetics?

Isolation by distance (IBD) has been a common measure of genetic structure among populations and is based on Euclidean distances among populations. Whereas IBD does not incorporate geographic complexity (e.g. dispersal barriers, corridors) that may better predict genetic structure, a new approach (landscape genetics) joins landscape ecology with population genetics to better model genetic structure. Should IBD be set aside or should it persist as the most simple model in landscape genetics? We evaluated the status of IBD by collecting and analyzing results of 240 IBD data sets among diverse taxa and study systems. IBD typically represented a low proportion of variance in genetic structure (mean r²=0.22) in part because many studies included relatively few populations (mean=11). The number of populations studied (N) was asymptotically related to IBD significance; a study with 9 populations has only 50% probability of significance, while one with >23 populations will have 90% probability of significance. Surprisingly, ectothermic animals were significantly (p=0.0018) more likely to have significant IBD than endotherms, which suggests a metabolic basis underlying gene flow rates. We also observed marginally significant effects on IBD significance for a) taxa in general and b) dispersal modes within actively‐dispersing endotherms. Other factors analyzed (genetic markers, genetic distances, habitats, active or passive dispersal, plant growth form) did not significantly affect IBD, likely related to typical N. For multiple reasons we conclude that IBD should continue as the simplest reference standard against which all other, more complex models should be compared in landscape genetics research.     

Cytokine/Chemokine HLDA8 Workshop panel report: analysis of receptors on lymphocytes from cord blood, normal and asthmatic subjects, and HIV positive patients

Chemokines and cytokines are involved in many processes, both physiological and pathological, particularly the recruitment, differentiation, activation, and proliferation of immune cells taking part in ontogenesis, inflammation, and cancer. It was assumed that chemokines and cytokines receptors are expressed in a regulated manner by human lymphocytes during ontogeny and later on, under the environmental stimulation of antigens they contribute to organogenesis, angiogenesis, and tissue remodeling, as well as modulating leukocyte effector functions. Using monoclonal antibodies classified by the Cytokine/Chemokine section of the 8th International Workshop on Human Leukocyte Differentiation Antigens, we analyzed human lymphocytes in blood samples drawn from the umbilical cord, normal adults, allergic and non-allergic asthma patients, HIV infected, and AIDS positive subjects. The main differences noted between adult and cord blood lymphocytes were related to CCR7 and CXCR4 receptors, which were more strongly expressed on cord blood lymphocytes, confirming the important role of these chemokines during development of the immune system. As with the HIV, CXCR4, and CCR5 co-receptors, we found no differences in CXCR4 expression between HIV and AIDS patients. However CCR5 was more strongly expressed in AIDS patients, which is likely to be associated with the evolution of disease. Further studies are needed to gain a better understanding of the functions of these molecules in the underlying pathogenesis of many diseases and to probe the use of the chemokine receptors as targets for therapeutic intervention.