The highly specialised gut of Fritillariidae (Appendicularia: Tunicata)

The morphology and some functions of the gut of Fritillaria pellucida and Fritillaria formica (Fritillaridae) were investigated by light and electron microscopy, and also by means of histochemical and immunohistochemical techniques. Fritillarids, very important for their abundance and ecological impact in marine ecosystems, have a very simplified gut: a straight oesophagus connects the pharynx to the digestive nucleus, composed of globular stomach and rectum, connected dorsally through a very short proximal intestine. The latter is characterised by a few (two to four) extremely specialised cells, completely filled with mitochondria associated tightly with membrane infoldings showing strong ATP-ase activity, and probably involved in the osmoregulation of internal body fluids. The gut is formed of an extremely low number of cells, which, although poorly diversified, are very large in the stomach and rectum. Food transfer along the gut depends on and is regulated by well-developed cardiac and pyloric valves, and signs of general digestive and absorptive activity are recognisable all along the brush border of the main tracts. The macroscopic organisation and cytological characters of the gut in fritillarids are completely different from those of the oikopleurids. In particular, fritillarids lack specialised cells for endocytosis and intracellular digestion, like those described in the genus Oikopleura. The general simplification and specialisation observed in Fritillaria gut may account for their elevated growth rate and abundant diffusion in all oceans.     

Release of phagocytosis-stimulating factor(s) by morula cells in a colonial ascidian

In vitro yeast phagocytosis by haemocytes of the compound ascidian Botryllus schlosseri was studied, with particular attention to interactions among different immunocyte types. It is demonstrated that the supernatant from haemocyte cultures matched with yeast cells contains factor(s) able to enhance yeast ingestion by Botryllus phagocytes. The increase in phagocytosis is not the consequence of yeast opsonisation, as the phagocytic index does not significantly increase when yeast cells, previously incubated in the culture media, are washed and re-suspended in filtered sea water. When haemocytes were fractionated by density gradient centrifugation and each band was incubated with yeast, the ability to stimulate phagocytosis was found in the supernatants from haemocyte cultures of fractions rich in morula cells (MC). Previous studies have demonstrated that MC express molecules recognised by anti-cytokine antibodies, as a consequence of the recognition of foreign molecules or cells. Our results indicate that molecules immunoreactive with anti-cytokine antibodies are required for modulating phagocyte activity, as the above-reported enhancing effect is completely absent in the presence of anti-IL-1-α and anti-TNF-α, but not of anti-rabbit-IgG antibodies, and they also highlight the presence of ‘cross-talk‘ between MC and phagocytes. A new scenario is therefore sketched, in which MC actively recognise non-self molecular patterns and, upon this recognition, release humoural factor(s) recognised by phagocytes, which modulate phagocytosis.     

Indicators for sustainable energy development: Brazil's case study

This article summarizes the results of the project on indicators for sustainable energy development (ISED) in Brazil. The project's aim was to present energy related economic, social and environmental data to policy makers in a coherent and consistent form, showing interlinkages, time‐series and cross‐sectoral analyses and assess energy policy. Two priority areas assessed by these indicators, regarding the country's energy supply and demand, helped in the identification of a number of energy policy options that focused on specific aspects of the country's energy sector. On the supply side, these options include the development and stimulation of renewable energy, such as small‐scale hydroelectric, wind, solar photovoltaic power and bagasse cogeneration; stimulation of programmes for ethanol use as automotive fuel and sugarcane bagasse cogeneration; and implementation of natural gas‐fired, combined heat and power (CHP) plants. On the demand side, policy options include: the full implementation of the law on efficiency standards for appliances; expansion of utility investment in end‐use energy efficiency; adoption of targets and protocols to reduce energy intensity in the industrial sector; improvement of passenger transport efficiency; and the creation of a fund to improve energy affordability for the poor.     

Immunotoxicity in ascidians: Antifouling compounds alternative to organotins – II. The case of Diuron and TCMS pyridine

Using short-term hemocyte cultures of the colonial ascidian Botryllus schlosseri exposed to various sublethal concentrations of Diuron (3-(3,4-diclorophenyl)-1,1-dimethylurea) and TCMS pyridine (2,3,5,6-tetrachloro-4-(metylsulphonyl)pyridine), we evaluated their immunotoxic effects through a series of cytochemical assays previously used for organotin compounds. At concentrations higher than 250 μ M and 10 μ M for Diuron and TCMS pyridine, respectively, both biocides exerted immunosuppressant effects on Botryllus hemocytes, causing i) deep changes in the cytoskeleton that irreversibly affect cell morphology and phagocytosis, ii) induction of DNA damage, iii) leakage of oxidative and hydrolytic enzymes due to membrane alteration. Unlike organotin compounds, Diuron and TCMS pyridine do not inhibit cytochrome-c-oxidase, and only TCMS pyridine triggers oxidative stress. When co-present, they exert an antagonistic interaction on cytoskeletal components.     

Multifarious activities of gut epithelium in an appendicularian (Oikopleura dioica: Tunicata)

The functions of the various gut regions of Oikopleura dioica (oesophagus, left and right gastric lobes, vertical intestine, mid-intestine and rectum) were investigated by means of histochemical, histoenzymatic and immunohistochemical techniques at light and electron microscopes. Ciliary food progression is evidenced by the presence of ATPases on cilia, along the entire gut, with the exception of the cardiac valve, a passive device controlling food direction. Absorptive processes (alkaline phosphatase), active transport (ATPases) and nitrogen excretion (D-amino acid oxidase) occur along the entire gut, in both ciliated microvillar and globular cells. The latter, typical of the left gastric lobe and rectum, are also involved in endocytotic processes (exogenous peroxidase as tracer) and intracellular digestion (!-amylase, aminopeptidase M, acid phosphatase, 5'-nucleotidase, non-specific esterase). The giant cells of the gastric band participate in extracellular digestion; they contain secretory granules positive to various hydrolytic enzymes, the activity of which is also recognisable in faecal pellets inside the intestinal lumen. Lipid storage occurs mainly in the right gastric lobe and vertical intestine, whereas protein storage takes place in the rectal granular cells. Epithelial transport and possible osmoregulation occur along the entire gut, especially at the level of diffuse baso-lateral interdigitations, which increase the plasmalemma surface enormously, are often associated with mitochondria and possess numerous ATPase pumps. Data extend previous histological observations and hypotheses on the physiological role of the various gut regions. The remarkable and specific location of enzymatic activities and nutrient storage are in agreement with the high capacity of O. dioica to process a great quantity of food very rapidly and efficiently.     

Irgarol inhibits the synthesis of ATP in mitochondria from rat liver

The interactions of Irgarol with rat liver mithocondrial have been investigated. The results indicate that Irgarol inhibits the ATP synthesis. The analysis of the various steps involved in the ATP synthesis suggests that the inhibition is due to the opening of small-size pores.     

Potential disruptive effects of copper-based antifouling paints on the biodiversity of coastal macrofouling communities

Environmental Science and Pollution Research - The expanded use of copper(I)-based antifouling paints (AF) has increased copper leaching into coastal environments, requiring attention and...     

Immunotoxic effects of organotin compounds in Tapes philippinarum

One of the most harmful groups of coastal pollutants is the organotin compounds (OTCs) which have severe effects on both aquatic organisms and mammals including humans. The immunotoxic effects of OTCs were studied in the cultivated clam Tapes philippinarum by determining the immunosuppressant role on in vitro yeast phagocytosis at low doses (0.01, 0.05, 0.1 μM). The phagocytic index was significantly reduced in an irreversible non-lethal manner depending on concentration and lipophilic affinity. The order of inhibition was TBT≥DBT>MBT for butyltins and TPTC>TPTA≥TPTH for triphenyltins.     

Immunotoxicity in ascidians: antifouling compounds alternative to organotins: III--the case of copper(I) and Irgarol 1051

After the widespread ban of TBT, due to its severe impact on coastal biocoenoses, mainly related to its immunosuppressive effects on both invertebrates and vertebrates, alternative biocides such as Cu(I) salts and the triazine Irgarol 1051, the latter previously used in agriculture as a herbicide, have been massively introduced in combined formulations for antifouling paints against a wide spectrum of fouling organisms. Using short-term (60 min) haemocyte cultures of the colonial ascidian Botryllus schlosseri exposed to various sublethal concentrations of copper(I) chloride (LC₅₀ = 281 μM, i.e., 17.8 mg Cu L⁻¹) and Irgarol 1051 (LC₅₀ > 500 μM, i.e., >127 mg L⁻¹), we evaluated their immunotoxic effects through a series of cytochemical assays previously used for organotin compounds. Both compounds can induce dose-dependent immunosuppression, acting on different cellular targets and altering many activities of immunocytes but, unlike TBT, did not have significant effects on cell morphology. Generally, Cu(I) appeared to be more toxic than Irgarol 1051: it significantly (p < 0.05) inhibited yeast phagocytosis at 0.1 μM (∼10 μg L⁻¹), and affected calcium homeostasis and mitochondrial cytochrome-c oxidase activity at 0.01 μM (∼1 μg L⁻¹). Both substances were able to change membrane permeability, induce apoptosis from concentrations of 0.1 μM (∼10 μg L⁻¹) and 200 μM (∼50 mg L⁻¹) for Cu(I) and Irgarol 1051, respectively, and alter the activity of hydrolases. Both Cu(I) and Irgarol 1051 inhibited the activity of phenoloxidase, but did not show any interactive effect when co-present in the exposure medium, suggesting different mechanisms of action.     

Immunotoxicity in ascidians: antifouling compounds alternative to organotins - II. The case of Diuron and TCMS pyridine

Using short-term hemocyte cultures of the colonial ascidian Botryllus schlosseri exposed to various sublethal concentrations of Diuron (3-(3,4-diclorophenyl)-1,1-dimethylurea) and TCMS pyridine (2,3,5,6-tetrachloro-4-(metylsulphonyl)pyridine), we evaluated their immunotoxic effects through a series of cytochemical assays previously used for organotin compounds. At concentrations higher than 250 micro M and 10 micro M for Diuron and TCMS pyridine, respectively, both biocides exerted immunosuppressant effects on Botryllus hemocytes, causing i) deep changes in the cytoskeleton that irreversibly affect cell morphology and phagocytosis, ii) induction of DNA damage, iii) leakage of oxidative and hydrolytic enzymes due to membrane alteration. Unlike organotin compounds, Diuron and TCMS pyridine do not inhibit cytochrome-c-oxidase, and only TCMS pyridine triggers oxidative stress. When co-present, they exert an antagonistic interaction on cytoskeletal components.