Patterns of enzyme activities in the gills of the catfish Heteropneustes fossilis (Bloch) exposed to the anionactive detergent Na-alkyl-benzenesulphonate (LAS)

The effect of sodium-alkyl-benzenesulphonate (LAS) on the activity of the respiratory enzymes of the gills of Heteropneustes fossilis (Bloch) was investigated. After 48 h exposure, the main injury to gills was the progressive separation of the lamellae from their vascular components. The enzymes of the aerobic part of the metabolism showed a decrease in activity, whereas the activity of lactate dehydrogenase was strongly increased, thus indicating that LAS has a high potential to interfere with aerobic mechanisms; however, the mode of action of it has yet to be clearly defined.     

Profiles of enzyme activity in larvae of two cyprinid species with contrasting life styles (Cyprinidae; Teleostei)

Growth and the development of gills, muscle fibres and 10 enzymes serving different metabolic functions were studied in larvae of Rutilus rutilus (L.) and Chalcalburnus chalcoides (Agassiz, 1832). R. rutilus starts swimming and feeding one to three days after hatching, whereas in C. chalcoides this process is delayed by about 10 days. This difference in behaviour is reflected in the time-course of growth, the differentiation of the red muscle fibres and the activity of the enzymes of aerobic energy metabolism. On the other hand, the activity of the enzymes of anaerobic energy metabolism increases steadily throughout the period of observation (up to 60 days post-hatch), this trend being more pronounced in C. chalcoides than in R. rutilus . A weight-independent and a weight-dependent phase of development can be distinguished in the enzymes of aerobic energy metabolism. It is suggested that, in accord with previous findings, the early phase of locomotor activity of cyprinid larvae is fuelled mainly by aerobic processes, and that the central muscle mass of the larvae is more aerobic than the white muscle fibres of the adults. The delayed development of aerobic enzyme activity in C. chalcoides is compensated by the accelerated development of anaerobic enzyme activity, particularly of creatine kinase and lactate dehydrogenase. This difference between the two species studied suggests differences in the metabolic basis of burst activity.     

Interactions between vascular actions of detergent and catecholamines in perfused gills of European eel, Anguilla anguilla L. and brown trout, Salmo trutta L.

Noradrenaline induced vasodilation in isolated perfused eel or trout gills was inhibited by propranolol, aβ-adrenergic blocking agent. The noradrenaline effect was much reduced in gills of fish which had been kept in 1 mg/l solutions of the detergent LAS (linear alkylate sulphonate; 12C chain length). LAS produced concentration-dependent vasodilation, which was inhibited by propranolol, in eel and trout gills. Noradrenaline produced additional vasodilation in eel and trout gills which had been dilated by 2.10⁻⁸ m LAS but had no effect on trout gills perfused with 2.10⁻⁷ m LAS. Gills from trout which had been kept in 0.1 mg l^-1 LAS responded to noradrenaline normally but the propranolol inhibition appeared to be potentiated.     

Biotransformation of linear alkylbenzene sulfonate (LAS) by Phanerochaete chrysosporium: oxidation of alkyl side-chain

The white rot fungus Phanerochaete chrysosporium, which generally mineralizes substituted aromatics to CO₂, transformed linear alkylbenzene sulfonate (LAS) surfactants mainly at their alkyl side chain. Degradation of LAS was evidenced by a zone of clearing on LAS-containing agar plates and colorimetric analysis of liquid cultures. Disappearance of LAS was virtually complete within 10 days in low nitrogen (2.4 mM N), high nitrogen (24 mM N) and malt extract (ME) liquid media. After 5 days of incubation in ME medium, transformation of LAS was complete at concentrations4 mg l^-1, but decreased at higher concentrations. The LAS degradation was not dependent on lignin peroxidases (LiPs) and manganese-dependent peroxidases (MnPs). Mineralization of¹⁴C-ring-LAS to ¹⁴CO₂ by P. chrysosporium was <1% regardless of the culture conditions used. Thin layer chromatography and mass spectral analyses indicated that P. chrysosporium transformed LAS to sulfophenyl carboxylates (SPCs) through oxidative shortening of the alkyl side-chains. While LAS disappearance in the cultures was not dependent on LiPs and MnPs, transformation of the parent LAS moieties to SPCs was more extensive in low N medium that favors expression of these enzymes. The SPCs produced in LN cultures were shorter in chain-length than those produced in ME cultures. Also there was a notable shift in the relative abundance of odd and even chain length metabolites compared to the starting LAS particularly in the low N cultures suggesting the possible involvement of processes other than or in addition to-oxidation in the chain-shortening process.     

Antioxidant defence enzyme activities in hepatopancreas, gills and muscle of Spiny cheek crayfish (Orconectes limosus) from the River Danube

The aim of our study was to determine the activity of antioxidant defence (AD) enzymes: superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR) and the phase II biotransformation enzyme glutathione-S-transferase (GST) in the hepatopancreas, the gills and muscle of Spiny cheek crayfish (Orconectes limosus) from the River Danube and to compare tissue specificities of investigated enzymes. Our results indicated that both specific and total SOD activities in the hepatopancreas were lower compared to the gills and muscle. Total SOD activity in the gills was lower with respect to that in muscle. CAT and GSH-Px (both specific and total) activities were higher in the hepatopancreas compared to those in the gills and muscle. In the gills the specific and total GR activities were higher than in the hepatopancreas and muscle. The specific and total GST activities were higher in the hepatopancreas compared with the gills and muscle. Our study represents the first comprehensive report of AD enzymes in tissues of O. limosus caught in the River Danube. The noted tissue distributions of the investigated AD enzyme activities most likely reflected different metabolic activities and different responses to environmental conditions in the examined tissues.     

直链烷基苯磺酸盐(LAS)的生物降解性

阴离子表面活性剂直链烷基苯磺酸盐（LAS）在工业生产和人们生活中得到了广泛应用,但大量产生的含有LAS的污水在环境中造成的污染也日益严重,因而引起了人们的高度重视,本从LAS的好氧和厌氧生物降解以及LAS降解菌等诸多方面阐述了LAS生物降解性的研究进展。     

Fate of linear alkylbenzene sulfonates in the environment: A review

The fate and risk of linear alkylbenzene sulfonates (LAS) in various compartments of the environment have been reviewed. Under aerobic conditions LAS degrade rapidly and concentration of LAS has been found very low in effluents from aerobic sewage treatment plants (STP). On the contrary, in anaerobic STPs effluents, LAS concentrations have been found very high. Anaerobic effluents containing high LAS concentrations have been found to pose risk to aquatic environment. Similarly, LAS concentrations have been found high in anaerobically treated sewage sludge's. LAS enter the soil as a result of sludge application to the land. Risk to aquatic and terrestrial ecosystems is increased when wastewaters and sludge's are treated anaerobically.     

Anaerobic biodegradation of linear alkylbenzene sulfonate (LAS) in upflow anaerobic sludge blanket (UASB) reactors

The anaerobic biodegradation of Linear Alkylbenzene Sulfonate (LAS) was studied in Upflow Anaerobic Sludge Blanket Reactors (UASB). One reactor was fed with easily degradable substrates and commercial LAS solution during a period of 3 months (Reactor 1), meanwhile a second reactor was fed with a commercial LAS solution without co-substrate (Reactor 2) during 4 months. Both reactors were operated with an organic loading rate of 4–5 mg-LAS/l*day and a hydraulic retention time of one day.The LAS biodegradation was determined by full mass balance. LAS was analysed by HPLC in the liquid phase (influent and effluent streams of the reactors) as well as in the solid phase (granular sludge used as biomass). The results indicate a high level of removal (primary biodegradation: 64–85%). Biodegradation was higher in the absence of external co-substrates than in the presence of additional sources of carbon. This indicates that the surfactant can be partially used as carbon and energy source by anaerobic bacteria. Under the operating conditions used, inhibition of the methanogenic activity or any other negative effects on the biomass due to the presence of LAS were not observed. The methanogenic activity remained high and stable throughout the experiment.     

Long-term experiment on Branchiura sowerbyi Beddard (Oligochaeta,Tubificidae) using sediment treated with LAS (Linear Alkylbenzene Sulphonate)

A long-term experiment was performed with Branchiura sowerbyi in order to assess possible effects of LAS sorbed to sediment on its reproductive cycle, using concentrations in sediment 2–5 times higher than those calculated for the LC50 values of LAS dissolved in water. No significant effects were observed during the whole experiment, so that we can confirm that LAS adsorbed on sediment has a much lower influence on the examined animals than LAS dissolved in water.     

Effect of cadmium on antioxidant enzyme activities and lipid peroxidation in the gills of the clam Ruditapes decussatus

Metals are known to influence the oxidative status of marine organisms, and antioxidant enzymes have been often proposed as biomarkers of effect. The clam Ruditapes decussatus is a well-known metal bioindicator. In this species cadmium (Cd) induces metallothionein (MT) synthesis only after 7 days of exposure. Before MT synthesis is induced, the other mechanisms capable of handling the excess of Cd are unknown. In order to identify some of these mechanisms, variations in antioxidant systems (superoxide dismutase, catalase, selenium-dependent glutathione peroxidase and non-selenium-dependent glutathione peroxidase), malondialdehyde (MDA) and MT were studied in the gills of R. decussatus exposed to different Cd concentrations (4, 40 and 100 gl^-1) for 28 days. These parameters, together with total proteins and Cd concentrations, were measured in the gills of the clams over different periods of exposure. Results indicate that Cd accumulation increased linearly in the gills of R. decussatus with the increase in Cd concentration. This increase induces an imbalance in the oxygen metabolism during the first days of Cd exposure. An increase in cytosolic superoxide dismutase (SOD) activity and a decrease in mitochondrial SOD activity was observed at the same time as or after a decrease in cytosolic and mitochondrial catalase activity and of selenium-dependent and non-selenium-dependent glutathione peroxidase activity. After 14 days of exposure, Cd no longer affect these enzymes but there was elevation of other cellular activities, such as MDA and MT production. MT bound excess Cd present in the cell. These variations in these parameters suggest their potential use as biomarkers of effects such as oxidative stress resulting from Cd contamination in molluscs.     

Branchial carbonic anhydrase (CA) of gills of Chasmagnathus granulata (Crustacea Decapoda)

The occurrence, localization and response to environmental salinity of carbonic anhydrase (CA) activity were studied in all of the gills of the euryhaline crab Chasmagnathus granulata from Mar Chiquita coastal lagoon (Buenos Aires Province, Argentina). CA activity in all gills appeared to be dependent on salinity. The pattern of distribution of CA activity among gills was different upon transition of C. granulata from osmoionoconformity (more uniform distribution) to hyperregulation (highest activity in posterior gills 6-8). Upon abrupt salinity change a differential response of CA activity occurred among gills which could suggest a differential role of CA in ion transport process in different gills of this crab. Furthermore, CA activity in anterior and posterior gills was found in cytosolic and microsomal fractions, although highest activity appeared to be membrane-associated. Both pools of CA were also strongly influenced by salinity and very sensitive to sulfonamide acetazolamide. The results suggest a differential participation of branchial CA in ionoregulatory mechanisms of C. granulata.     

ω-Oxygenation of the alkyl sidechain of linear alkylbenzenesulfonate (LAS) surfactant in <Emphasis Type="Italic">Parvibaculum lavamentivorans</Emphasis><Superscript>T</Superscript>

Parvibaculum lavamentivorans ^T DS-1, an aerobic, heterotrophic bacterium, requires a biofilm on a solid surface (e.g. glass particles) when utilizing commercial linear alkylbenzenesulfonate surfactant (LAS; 20 congeners) for growth. Catabolism involves the undefined ‘ω-oxygenation’ and β-oxidation of the LAS side chain, and the organism excretes sulfophenyl carboxylates (SPC) quantitatively. A 3.5-l fermenter was developed which allowed gram-quantities of LAS-grown cells to be grown and harvested from medium with glass particles as the solid support. The catabolism of LAS was dominant: in diauxie experiments with acetate as second carbon source, LAS was utilized first. The biofilm-encoated LAS-grown cells were unsuitable for metabolic work in vitro because cell suspensions clumped and were not disrupted effectively, but the degradative enzymes were found to be expressed constitutively in acetate-grown cells, which formed no biofilm. LAS-dependent oxygen uptake was measured in acetate-grown cells at about 0.6 mkat (kg protein)⁻¹, but not in extracts of cells. Whole cells converted LAS to SPC in the presence of molecular oxygen only, and the reaction could be saturably inhibited by metyrapone, which acts on e.g. cytochromes P450 (CYP). However, despite the presence of CYP153-like sequences in the genome of strain DS-1^T, the difference spectra did not support the presence of a CYP in crude extracts, and the nature of the LAS-oxygenase remains unclear.     

Effect of aerial exposure on the antioxidant status in the subantarctic stone crab Paralomis granulosa (Decapoda: Anomura)

In Tierra del Fuego (Southern South America), the stone or false king crab, Paralomis granulosa represents one of the most important crab fisheries. After capture, animals are kept in baskets and exposed to dryness for several hours, when the water flow through the gills is interrupted. As a consequence a concomitant increase of reactive oxygen species begins, triggering oxidative stress. The aim of this study was to determine oxidative stress and antioxidant enzyme activities due to air exposure in different tissues of P. granulosa. Fifty crabs (carapace length >82 mm) were captured in Beagle Channel (54 degrees 50'S, 68 degrees 20'W) during winter 2004. Five groups of 10 crabs each were exposed to dryness at 6 degrees C for 0, 3, 6, 12 or 24 h, respectively. Activity of superoxide dismutase (SOD), catalase (CAT), glutathione S transferase (GST) protein and lipid oxidation were measured in gills, muscle, hepatopancreas and haemolymph samples. Almost all analyzed tissues showed antioxidant enzymes activity, which varied with time of air exposure. The maximum enzyme activity was measured after 6 h of air exposure. Protein oxidation levels varied significantly in gills. Lipid peroxidation levels increased significantly in muscle and hepatopancreas. The critical time of air exposure probably occurs at 6 h. Thereafter animals were unable to induce the synthesis of antioxidant enzymes or proteins. This should be taken into account to minimize the stress generated by the commercial capture process.     

Acute toxicity of nitrite in fat snook (Centropomus parallelus)

Oxidative stress and hypoxia responses in Centropomus parallelus exposed to three different concentrations of nitrite (0, 0.5, and 10 mg L^?1) were evaluated. To determinate toxicity, we used enzymatic analyses and quantifications of oxygen consumption and opercular beats under conditions of normoxia and hypoxia. There was no significant difference in GST, CAT, or GPx enzymes in the liver or in GPx in the gills. Levels of GST were altered at a concentration of 10 mg L^?1 in experiment I. Experiment II displayed alterations for GST and CAT in the liver at the same concentration, indicating that exposure to nitrite does not generate differentiated activation in the body’s defense system by the production of oxidative stress enzymes. When nitrite was associated with hypoxia, there was a significant increase in both enzymatic activity and opercular beating under hypoxia in the group exposed to 10 mg L^?1 nitrite compared to the control.     

First evidences of surfactant biodegradation by marine sponges (Porifera): an experimental study with a linear alkylbenzenesulfonate

The main objective of this work was to better appreciate the role of benthic macro-organisms in the biodegradation of detergents in the marine environment. Sponges, which could be highly resistant to pollution and which are highly active filter-feeders, appear as interesting organisms in this topic. An experimental study conducted in aquarium with seawater enriched in a pure linear alkylbenzenesulfonate (LAS), namely 1-(p-Sulfophenyl)nonane, has shown that the primary degradation was 10 times more rapid in the presence of the sponge Spongia officinalis than in the presence of only marine bacteria. The main degradation metabolites, 3-(p-sulfophenyl)propionic acid and p-sulfobenzoic acid were produced in greater amounts in the presence of sponge. The very rapid degradation kinetics observed in this study may be due to the symbiotic microbial community present in S. officinalis. The sponge may also have promoted the activity of marine bacteria by transforming the original molecule in a more available secondary product. This study demonstrates for the first time that benthic macro-organisms can play an important role in the transformation of biodegradable contaminants, such as LAS.     

Effects of extracts from the cyanobacterium Microcystis aeruginosa on ion regulation and gill Na+,K+-ATPase and K+-dependent phosphatase activities of the estuarine crab Chasmagnathus granulata (Decapoda,Grapsidae)

Recent discoveries indicate that microcystins affect enzymes, such as Na(+),K(+)-ATPase, involved in ion regulation of aquatic animals, through K(+)-dependent phosphatase inhibition. In vitro studies showed the inhibitory effect of Microcystis aeruginosa extracts on Na(+),K(+)-ATPase and K(+)-dependent phosphatase activities in gills of Chasmagnathus granulata (Decapoda, Grapsidae). Extracts of M. aeruginosa were prepared from lyophilized or cultures cells of the cyanobacterium. For lyophilized cells, IC(50) values were estimated as 0.46 microg/L (95% confidence interval [CI]=0.40-0.52 microg/L) and 1.31 microg/L (95% CI=1.14-1.51 microg/L) for Na(+),K(+)-ATPase and K(+)-dependent phosphatase, respectively. However, extracts prepared from cultured cells presented a much lower inhibitory potency against both enzymes. Gas chromatography revealed long-chain fatty acids in the lyophilized cell extracts, indicating that they are in part responsible for the enzyme inhibition. In vivo studies showed that the toxin inhibited Na(+),K(+)-ATPase activity in anterior gills, whereas an increased augmented activity of glutathione-S-transferase was observed in both kind of gills, indicating that the crab has increased its ability to conjugate the toxin. No significant differences in hemolymph sodium or chloride concentration were detected. This result is in agreement with the lack of effects of microcystin on Na(+),K(+)-ATPase activity of posterior (osmoregulating) gills.     

The morphology of gills of Haliotis tuberculata (Linnaeus, 1758)

Manganaro, M., Laurà, R., Guerrera, M.C., Lanteri, G., Zaccone, D. and Marino, F. 2011. The morphology of gills of Haliotis tuberculata (Linnaeus, 1758). —Acta Zoologica (Stockholm) 93 : 436–443. Although the morphology of abalone gills has been studied by some authors, up to date no data are available about the gills of Haliotis tuberculata. This study was carried out, by light and electron microscopy, on 10 wild adult H. tuberculata. Gills lamellae produce an undulated surface increasing the area in contact with water. At the level of skeletal rods, we observed a joint‐like structure that allows a checked movement. The left ctenidium is always decidedly larger than the right, probably because of the enormous size of the shell muscle. The cilia permit oxygenated water that leaves the afferent border and is thrust away at the tips of the lamellae by the extremely long cilia. Ciliary movement may take part in sweeping mucous secretions to capture extraneous particles and remove them from the gills. Three types of mucous cells are distributed along the epithelium of the afferent and efferent zones of the gill filament. They seem to play a role in the cleansing of gills in coordination with the muscle contraction and ciliary movement. The presence of microvilli on particular cells reflects their role associated with the absorption of substances from the environment. A haemolymphatic vessel is located in the central zone of the gill filament. The backbone of the haemolymphatic vessel is a chitino‐like structure, which gives support to the gills.     

A new species of Isocyamus (Amphipoda: Cyamidae) from Kogia breviceps (De Blainville, 1838) in Australian waters

Isocyamus kogiae n. sp. is described from a pygmy sperm whale Kogia breviceps (De Blainville, 1838) stranded on Moreton Island, Queensland, Australia. It belongs in the genus Isocyamus because of its white appearance, epaulette-like processes at the anterior dorsum of pereon segment 2, lateral spines on segments 3 and 4, reduced maxilliped, and absence of maxilliped palps, large accessory gills and dentate comb on the dactyle of gnathopod 1. It differs from Isocyamus delphini (Guerin-Meneville, 1836), the only other member of this genus, in that the accessory gills are not equal in length to the main gills, it has no ventral spines, and the dactylus of percopod 1 has a double rather than a single dentate comb. This is the first report of a cyamid from whales of the genus Kogia.     

Effects of environmental anoxia and different periods of reoxygenation on oxidative balance in gills of the estuarine crab Chasmagnathus granulata

We investigated the effects of anoxia (8 h) and different periods of reoxygenation (20 and 40 min) on the oxidative balance in anterior and posterior gills of the crab Chasmagnathus granulata. Enzyme activity of catalase and GST was increased in the gills of the animals submitted to anoxia, and SOD activity was decreased. These enzymes returned approximately to control levels during the anoxia recovery time. These results demonstrated enzyme activities change with variations in environmental oxygen levels. The posterior gills showed a higher antioxidant enzyme activity than anterior gills. In the gills, there were no changes in the non-enzymatic antioxidant system (TRAP) during anoxia. On the other hand, during anoxia recovery, an increase of TRAP in both gills was observed. Anoxia does not change lipid peroxidation (TBARS) in the gills. During anoxia recuperation, an increase in levels of TBARS was observed. Thus the results demonstrate that C. granulata has a similar strategy of preparation for oxidative stress as observed in other intertidal species, enabling the crabs to survive in an environment with extreme variations in physical and chemical characteristics, such as salt marshes.