Aroclor 1254 inhibits the chemiluminescence response of peritoneal cavity cells from sharpsnout sea bream (Diplodus puntazzo)

Chronic exposure to polychlorinated biphenyls (PCBs) affect the immune system of fish and could lead to a decreased disease resistance. The effects of Aroclor 1254, PCB mixtures, on the Diplodus puntazzo innate immunity were examined by assaying the zymosan stimulated chemiluminescence response (CL) of peritoneal cavity cells (PCCs) at various times (1, 24, 48 h and 1-4 weeks) from intraperitoneal injection of the xenobiotic (1 mg kg(-1) body weight). Controls were performed by assaying cells from medium-treated fish. Since the kinetic of the chemiluminescence response showed the highest peak at 25 min after the zymosan stimulation of the cells, the values found at that time were considered. The CL enhancement observed at 1 h after the treatment with xenobiotic was followed by a decreased response at 24 h and appeared to be lower at 1-4 weeks when compared to the CL response of the control, suggesting a protracted effect of PCBs on the peritoneal cavity. Since PCCs incubated in vitro for 1 h with 0.05 and 0.1 mu g ml(-1) Aroclor showed an enhanced CL, the effect of the xenobiotic could be exerted on the cell responsiveness to zymosan. It is known that fish CL response of PCCs can be imputed to phagocyte (macrophages and neutrophils) activation, these cells and their responsiveness to zymosan can be used in immunotoxicology assay to monitor the fish health in polluted environment. (C) 2014 Elsevier Ltd. All rights reserved

Differential expression of two glucocorticoid receptors in seabass (teleost fish) head kidney after exogeneous cortisol inoculation

Stressful conditions include a prompt release of corticosteroid hormones which can mediate gene expression through glucocorticoid receptors (GR). Since two seabass (Dicentrarchus labrax) GRs have been cloned and sequenced from peritoneal cavity cells (DlGR1) and liver (DlGR2), a comparative amino acid sequence analysis that included Haplochromis burtoni HbGRs, was carried out and homologies disclosed. The DlGR1 and DlGR2 deduced aminoacid sequences showed 61% identity (I) and 70% similarity (S). Moreover, DlGR2 was similar to HbGR2b (69% I, 73% S), and the DlGR1 to HbGR1 (72% I, 78% S). In addition, we examined the expression of the DlGRs after exogeneous cortisol inoculation into the peritoneal cavity, mimicking stress effects. At various times after the administration (3 h, 24 h, 1 week), gene expressions was evaluated in head kidney by real-time PCR. In addition, immunoblotting and densitometry analyses were performed with anti- DlGR1 antibodies. Although sea bass head kidney expressed both DlGR1 and DlGR2 they were differentially modulated by intraperitoneal implant of exogeneous cortisol

Effects of diclofenac on the gametes and embryonic development of Arbacia lixula

Diclofenac (DCF), a non-steroidal anti-inflammatory drug (NSAID), is among the main pollutants of aquatic environments. Nevertheless, even if several authors evaluated its effects on marine organisms, no work has ever analysed its impact on the sea urchin Arbacia lixula. The purpose of this study was to analyse, for the first time, the impact of DCF at different concentrations (50 mg/l, 5 mg/l, 0.5 mg/l, 0.05 mg/l) on A. lixula embryos and gametes to indicate possible safe minimum levels of release of this drug in marine environments. Our results showed significant concentration-dependent effects on embryonic development (decrease in the number of developed embryos and increase in the quantity of degenerated eggs) and significant levels of bioaccumulation at DCF concentrations of 50 mg/l in both gametes and embryos. DCF showed a significant impact on the fertilisation of A. lixula due to its effects on gametes, confirming the possibility of environmental risk and highlighting the need to improve wastewater treatment and drug disposal processes

Expression and distribution of the glucocorticoid receptor DlGR1 in the teleost Dicentrarchus labrax brain.

Cortisol is the main corticosteroid secreted by the interrenal cells of the head kidney and it exerts a role in mantaining the omeostatic status in fish. In teleosts its effects are mediated through intracellular receptors expressed in several tissues, that are ligand-dependent transcription factors by binding to specific tissue DNA sequences. In Dicentrarchus labrax we previously cloned and sequenced a glucocorticoid receptor, DlGR1, isolated from leukocytes of peritoneal cavity. In this work we showed mRNA expression and tissue immunohistochemical localization of brain DlGR1 by in situ hybridization assays, with a riboprobe with DlGR1 cDNA trascriptional activation domain, and by immunohistochemical methods, using a specific antibody for a selected sequence of the receptor tran- scriptional domain. The mRNA and the protein are expressed in pyramidal cells of the optic lobe and in the small globular neurons of the diencephalon

Serum thyroid hormone evaluation during transition periods in dairy cows

Abstract. Thirty-five multiparous Holstein dairy cows were selected from a high-producing dairy farm in northeastern Italy: 16 in second lactation (L2), 10 in third lactation (L3) and 9 in fourth lactation (L4). Blood sampling was carried out 7 ± 5 days before calving (Pre/C) and 7 ± 5 days after calving (Post/C). Serum triiodothyronine (T3), thyroxine (T4) and a thyroid-stimulating hormone (TSH) were assessed. Two-way analysis of variance (ANOVA) showed statistically significant effects of class (L2, L3, L4) and of period (Pre/C, Post/C) on the parameters studied (T3, T4, TSH). In particular, Bonferroni's multiple comparison test showed lower values in post-calving than in the pre-calving in L2 and L3 for TSH; lower values in post-calving than in the pre-calving in L2 for T3; and lower values in post-calving than in the pre-calving in L2, L3 and L4 for T4. Our results improve the knowledge of endocrine and metabolic changes occurring in dairy cows during transition periods and may be useful to supply a new strategy for the improvement of dairy cow farm management and reproductive performance

Behavioural and biochemical stress responses of Palinurus elephas after exposure to boat noise pollution in tank

This study examined the effects of boat noise on the behavioural and biochemical parameters of the Mediterranean spiny lobster (Palinurus elephas).The experiment was conducted in a tank equipped with a video and audio recording system. 18 experimental trials, assigned to boat noise and control conditions, were performed using lobsters in single and group of 4 specimens. After a 1. h habituation period, we audio- and video-recorded the lobsters for 1. h. During the experimental phase, the animals assigned to the boat groups were exposed to boat noise pollution (a random sequence of boat noises). Exposure to the noise produced significant variations in locomotor behaviours and haemolymphatic parameters. Our results indicate that the lobsters exposed to boat noises increased significantly their locomotor activities and haemolymphatic bioindicator of stressful conditions such as glucose, total proteins, Hsp70 expression and THC when tested both singly and in groups. © 2014 Elsevier Ltd

Assessing the potential of INFFER to improve management of agro-environmental assets in Tuscany

In the European Union, as in many parts of the world, there is increasing pressure to demonstrate that environmental projects achieve expected outcomes and are cost-effective. This paper reports on a trial application of an environmental investment framework, the Investment Framework for Environmental Resource (INFFER). The framework, based on benefit:cost analysis principles, was applied to agro-environmental assets in the Mugello and Valdisieve areas of northern Tuscany, Italy. Farmers, environmental and institutional representatives and technical experts took part in the process of identifying agro-environmental assets in the region. A total of 26 assets were identified, and of these, only 2 (the last remaining population of Cistus laurfolius within the natural protected area of local interest ( Area Naturale Protetta di Interesse Locale , ANPIL) of Santa Brigida, and the Sant' Antonio forest were selected as having reasonable prospects of progressing to cost-effective projects (benefit:cost ratio>1). Detailed projects were developed for both assets and found to be cost-effective based on available information. While the mindset of being clear about identifying and valuing natural assets was a new concept to participants, overall the process worked well. The factors considered in calculating a benefit:cost ratio, including technical feasibility, adoption and compliance factors, socio-political risks, costs and the likelihood of obtaining long-term funding to maintain project benefits, were all relevant in Tuscany and required the knowledge of local experts. Overall, INFFER appears relevant and useful in an EU context, and, in particular, is highly compatible with the EU Natura 2000 nature and biodiversity policy

In Vitro Cytotoxic Effect of Aqueous Extracts from Leaves and Rhizomes of the Seagrass Posidonia oceanica (L.) Delile on HepG2 Liver Cancer Cells: Focus on Autophagy and Apoptosis

Aqueous extracts from Posidonia oceanica’s green and brown (beached) leaves and rhizomes were prepared, submitted to phenolic compound and proteomic analysis, and examined for their potential cytotoxic effect on HepG2 liver cancer cells in culture. The chosen endpoints related to survival and death were cell viability and locomotory behavior, cell-cycle analysis, apoptosis and autophagy, mitochondrial membrane polarization, and cell redox state. Here, we show that 24 h exposure to both green-leaf- and rhizome-derived extracts decreased tumor cell number in a dose– response manner, with a mean half maximal inhibitory concentration (IC50) estimated at 83 and 11.5 µg of dry extract/mL, respectively. Exposure to the IC50 of the extracts appeared to inhibit cell motility and long-term cell replicating capacity, with a more pronounced effect exerted by the rhizomederived preparation. The underlying death-promoting mechanisms identified involved the downregulation of autophagy, the onset of apoptosis, the decrease in the generation of reactive oxygen species, and the dissipation of mitochondrial transmembrane potential, although, at the molecular level, the two extracts appeared to elicit partially differentiating effects, conceivably due to their diverse composition. In conclusion, P. oceanica extracts merit further investigation to develop novel promising prevention and/or treatment agents, as well as beneficial supplements for the formulation of functional foods and food-packaging material with antioxidant and anticancer propertie