Effects of salinity on the clam <Emphasis Type="Italic">Chamelea gallina</Emphasis> haemocytes. Part II: Superoxide dismutase response

The effects of different salinity levels (28, 34 and 40‰) on functional responses of Chamelea gallina haemocytes were evaluated in a two part study dealing with modulations of immune parameters. This part (Part II) of the study was focused on the superoxide dismutase (SODs) activity and expression in haemocyte lysate and cell-free haemolymph. Results of this study established that the exposure of C. gallina specimens at 40‰ salinity provoked a decrease in Mn-SOD and Cu/Zn-SOD activities in haemocyte lysate suggesting a declining superoxide anion generation at the highest salinity tested. Expression of MnSOD was coherent with activity values, while Cu/ZnSOD showed two immunoreactive bands. The former corresponds to the cytosolic Cu/Zn-SOD (16 kDa) was not coherent with the enzyme activity and the second (28–30 kDa) probably attributed to EC-SOD. In cell-free haemolymph, Mn-SOD activity decrease and Cu/Zn-SOD activity increase at 40‰ were observed, likely due to EC-SOD contribution, strongly induced at the same salinity. After EC-SOD detection with two different antibody, we postulate that the EC-SOD like-protein band (29 kDa) may be constitute partly by EC-SOD and probably by Cu/Zn-SOD dimeric form not completely dissociated under reducing condition or a Cu/Zn-SOD degenerated but still recognized by antibody. The cell-free haemolymph increase of EC-SOD at high salinity values plays an important role in immune defence of C. gallina. According to the conclusion of Part I of this study, our data confirmed the destabilizing effect of 40‰ salinity on haemocyte functionality, while to 28‰ exposure, data don’t confirm its stressful action as instead stated by results of Part I. Further studies are necessary to clear up this discrepancy.     

Effects of salinity on the clam <Emphasis Type="Italic">Chamelea gallina</Emphasis>. Part I: alterations in immune responses

In the present study, the effects of differing salinities on some important functional responses of haemocytes from the clam, Chamelea gallina, were investigated. The animals were kept for 7 days at 28‰ (hyposalinity), 34‰ (control) and 40‰ salinity (hypersalinity), and total haemocyte count (THC), haemocyte volume, phagocytosis, lysozyme-like activity (in both haemocyte lysate and cell-free haemolymph) were measured. The survival-in-air test was also performed. Clams kept at 28‰ showed significantly increased THC with respect to animals kept at 34 and 40‰. The analysis of haemocyte size frequency distribution highlighted that in clams kept at 28‰ the haemocyte fraction of about 5 μm in diameter and 50–100 femtolitre in volume increased markedly. Conversely, in animals kept at 40‰ an increase was observed in the haemocyte fraction having about 8–10 μm diameter and 400–500 femtolitre volume. Higher phagocytic activity was recorded in haemocytes from control clams, with respect to that of clams kept at 28 and 34‰. Lysozyme-like activity in haemocyte lysate was shown to increase significantly in animals kept at 28‰ with respect to that of clams kept at 40‰, whereas enzyme activity in cell-free haemolymph from clams kept at 34‰ was significantly higher with respect to that of clams maintained at 40‰. A relationship between phagocytosis and lysozyme secretion is suggested. The resistance to air exposure of clams kept at 28 and 40‰ was shown to decrease significantly; LT₅₀ values fell from 7 days in clams kept at 34‰ to 4 and 5 days in those kept at 28 and 40‰, respectively. Results demonstrated that salinity values far from 34‰ affects the functional responses of haemocytes and reduce the resistance of clams to exposure to air.     

Meiofauna densities and gastrotrich community composition in a Mediterranean sea cave

During the past 5 decades, the large-sized biota inhabiting dark marine caves has attracted the attention of many marine biologists; in contrast, studies concerning the meiofaunal organisms of these peculiar biotopes remain scanty and mostly with a taxonomic aim. In this study, the nature and abundance of meiofaunal taxa living in a Mediterranean, semi-submerged sea cave was surveyed in relation to distance from the entrance and over two different seasonal periods. Particular attention was paid to the Gastrotricha taxocene. Research was carried out in a cave along the Ionian coast of Apulia (southern Italy), the “Grotta Piccola del Ciolo” which opens for approximately 120 m on the north-eastern side of a shallow fjord and has a bottom blanketed by fine to very fine sand, occasionally rich in detritus. Quantitative samples in four replicates were collected by SCUBA diving, in November 2000 and June 2001, coring the sediment with a hand-held piston corer in three light-free sites (stations 1–3) located at increasing distances from the entrance. At each site, two additional 500-ml sediment samples were collected for an in vivo study of the Gastrotricha. Faunistic analysis found a fairly high meiobenthic diversity, identifying representatives of more than 12 major groups, with total abundances ranging from 656 ind./10 cm² (10 cm²) in November to 1,069 ind./10 cm² in June. Station 1, the closest to the entrance invariably hosted the most abundant meiofaunal community (851 ind./10 cm² in November and 1932 ind./10 cm² in June), followed by station 2 or 3 depending on the season. While nematodes and harpacticoids appear as the most abundant taxa when the cave is considered as a whole, other taxa may prevail numerically in selected stations, e.g. priapulids, which are the second most abundant taxon at station 1 (30 ind./10 cm² in November and 83 ind./10 cm² in June). Although the density of total meiofauna and that of the single groups may not be very high, the cave is interesting by virtue of the peculiarity of the hosted fauna, e.g., species and genera new to science or new to the Mediterranean Sea. Regarding the Gastrotricha, we found 16 species, accounting for 1.3–2.6% of the total meiobenthos (density = 8.4 ind./10 cm² in November and 27.4 ind./10 cm² in June). Analysis of the gastrotrich community found, particularly in June, an assemblage of taxa quite different from those found in open habitats, even at the family level; differences that are probably due to the exploitation of different food resources by animals populating the two environments, i.e. algae in the open sea versus bacteria in the caves. Results indicate that for meiofauna, as happens for macrofauna, the marine caves may represent hotspots of biodiversity and endemism; the driving forces at the base of the trophic depletion hypothesis seem to be responsible for structuring the meiofauna community inside the cave.     

Do male barn swallows (<Emphasis Type="Italic">Hirundo rustica</Emphasis>) experience a trade-off between the expression of multiple sexual signals?

Some sexual selection models envisage exaggerated male secondary sexual characters to be costly and therefore reliable indicators of the quality of potential mates to choosy females. If male secondary sexual characters have a natural selection cost, they may be linked to each other by reciprocally constraining relationships that would prevent individual males from increasing their level of multiple signaling. Barn swallow (Hirundo rustica) males have at least two costly signals relevant to socio-sexual interactions: tail length and song. Under the hypothesis that a trade-off exists between male signals, we manipulated the maintenance cost of tail ornaments to test whether this reduced the quantity and quality of song, a condition-dependent, phenotypically plastic signal. Contrary to our expectation, tail elongation had no effect on singing activity and song complexity. However, tail-elongated males produced songs with longer terminal parts ('rattles'). Long rattles are associated with highly competitive social contexts and high circulating levels of testosterone, suggesting that tail-elongated males were more frequently involved in either male-male aggressive or inter-sexual interactions. Therefore, this study shows that males are not displaying multiple signals at the maximum possible level, implying that this system is open to unreliable communication. However, long-term trade-offs between signal expression and viability may prevent males from displaying both signals at higher rates.Communicated by: M. Webster     

Effects of female mating status on copulation behaviour and sperm expenditure in the freshwater crayfish <Emphasis Type="Italic">Austropotamobius italicus</Emphasis>

Sperm competition models predict that males should adjust their sperm expenditure according to the risk and/or intensity of sperm competition. In this paper, we analysed copulatory behaviour of both sexes and sperm expenditure in relation to female mating status (virgin or mated) in the freshwater crayfish Austropotamobius italicus, a species where males have been reported to feed on and remove sperm laid by other males. The same females were allowed to be inseminated sequentially by two males, and we compared the sexual behaviours of partners between the first (virgin females) and the second mating (mated females). We found that female resistance did not differ between the first and the second mating, nor males refused or took more time to mount a mated female. However, when mating with a mated female, males reached an effective copulation position significantly later. This occurred because second-mating males removed, by eating, all or most spermatophores previously deposited by first males. As removal was often incomplete, this resulted in a larger amount of sperm being deposited on female ventral parts after the second mating, although second males did not allocate more sperm to mated females than first males did. Thus, the peculiar mode of sperm competition, where males remove previously deposited sperm, and the consequent predictable strong last male prevalence in paternity likely led to the observed lack of adjustment of sperm expenditure to female mating status in this species.     

Elemental Composition of Urban Aerosol Collected in Florence, Italy

An extensive investigation is in progress aiming at the characterisation of the air particulate composition in Florence. For our investigation, we use the external PIXE-PIGE beam facility of the I.N.F.N. Van de Graaff accelerator at the Physics Department of the Florence University. In order to gather information on both the longer- and shorter-time trends of the aerosol elemental composition, we are analysing both long temporal series (about 1 year) of 24-h Millipore filters collected by the health authorities in 3 different sites, and filters collected simultaneously in two of the above sites for about one month, by two streakers with one-hour resolution, providing size fractionation between particle size smaller than 2.5 µm and from 2.5 µm to 10 µm. The streakers sampling period includes two days during which the Municipality of Florence has banned the circulation of non-catalytic cars, due to the increase of NO₂ above the "recommended safety values". We present here the first obtained results for the sampling site located near a heavy traffic road.     

Analytical characterization of movements of the spinal column and risk assessment due to repeated movements of the upper limbs of building painters

Manual activities of construction workers may induce musculoskeletal disorders. This study on a group of painters aimed to analytically characterize movements of the spinal column by both lumbar motion monitor and television cameras and to determine, using the Occupational Repetitive Actions (OCRA) Index method, the risk exerted by repeated movements of the upper limbs. The main results are: painting with a roller generally exposes workers to a lesser risk for upper limbs than painting with a brush; a roller-stick fixed at the wrong length can lead to stretching of the back at lumbar and cervical levels; to remain within the range of ‘acceptable risk’ (OCRA Index evaluation), a worker should not paint a vertical wall for over 3 h if using a roller and 2.5 h if painting with a brush; and, on average, a painter who paints for 5 h in a day lifts the bucket about 120,140 times.     

Tensile Behavior of Thermoplastic Films from Wheat Flours as Function of Raw Material Baking Properties

The production of bioplastics directly from wheat flour has been demonstrated to be reliable, but scarce knowledge is available on how flour characteristics may affect the performance of thermoplastic films. In this work, we first established the most suitable recipe and process for the production of extruded films and then we used eight single-cultivar wheat flours with different baking technological properties to assess how they affect the mechanical properties of thermoplastic films. The results have shown that flours from soft grain cultivars offered more rigid and deformable films than flours from hard grain cultivars. For similar hardness, the alveographic P/L ratio of the dough was inversely related to rigidity and directly related to deformability of plastic films, while the deformation energy of the dough (W) played a role only for great differences of it. The subsequent fabrication of blends between each of the flours that yielded the best film properties and polycaprolactone (PCL) at different proportions indicated that a wheat flour/PCL ratio (TWF/PCL) of 75/25 offered the most suitable films for further application. Our results are likely to be useful for improving the plasticization of flour, in that selection of wheat flours could be tailored on the properties desired for the bioplastic films.     

Determination of estrogenic activity in the river Chienti (Marche Region, Italy) by using in vivo and in vitro bioassays

Environmental estrogen-like compounds (i.e. xenoestrogens) are a variety of pollutants, ranging from synthetic to natural occurring molecules, that are found in surface and waste waters over a wide range of concentrations. In aquatic environment, the overall estrogenic activity is often due to the presence of a mixture of chemicals and their degraded products which can induce synergistic effects. Current strategies for monitoring estrogen-like chemicals are based on the use of a battery of in vivo and in vitro ecotoxicological tests. In this regard, the aim of the present work was to carry out a bio-monitoring study for testing estrogenicity of the Chienti river (Marche Region, Italy) by using both an E-screen and a vitellogenin (Vtg) induction assay in juvenile goldfish. Three sites were used for analysis, localized at the mouth (sampling point 1), in the middle (sampling point 2) and at the origin (sampling point 3) of Chienti river. For most of the water samples (i.e. samples collected at sampling points 2 and 3), clear estrogenic activity was detected in the E-screen assay suggesting different proliferation activities in function of the collecting site. In contrast, the Vtg ELISA demonstrated that water samples collected from each sampling point were estrogenic. Overall, we showed for the first time that the estrogenic activities in water samples from the Chienti river were significant in both in vivo and in vitro; we also observed a different sensitivity between bioassays.