Physiological and biochemical studies of fish functional homeostasis

Stages of formation and development of physiological and biochemical studies of fish in Russia have been described. Main directions of studies and biochemical markers used for the assessment of the functional state of fish in natural populations and aquaculture have been described.     

Biodiversity in the Black Sea: effects of climate and anthropogenic factors

The Black Sea ecosystem and diversity underwent dramatic adverse changes during the 1960s and, especially, 1970s and 1980s of the last century. Anthropogenically-induced eutrophication increased through greater biogenic flow, dumping and pollutant discharge, in turn causing red tides, fish kills and oxygen depletion over the northwestern shelf. Anthropogenic pressures, associated with the economic situation of the Black Sea countries, has decreased during the last decade, allowing some improvement in the state and biodiversity of the ecosystem. The abundances of several native species have increased. However, mediterranization—the invasion by species from the adjacent basin and beyond—has continued. The conclusion is grounded, that biodiversity is not only inter- and intra-species diversity but also spatial–temporal variability, abundance and productivity dynamics, differences of the metabolic strategies providing sustainable existence in the changing environment. Biodiversity at the intra-species level expresses itself in spatial and temporal variations of the Black Sea biota. It has been shown, that рreservation of the Black Sea ecosystem’s biodiversity must be based on the measures which should be undertaken in national and social spheres, and be directed to the recreation, stabilization and conservation of this unique sea basin. Measures must be implemented nationally to conserve, stabilize or recreate the ecosystem biodiversity of this unique sea basin.     

结合代谢组学和转录组学分析恶臭假单胞菌Y-9主动稳定胞内外pH的机制

【目的】揭示恶臭假单胞菌(Pseudomonas putida) Y-9在氨氧化过程中主动调节胞外和胞内pH稳态机制。【方法】在初始pH为7.19和9.40的硝化培养基中培养Y-9生长48 h，利用代谢组学对比分析Y-9氨氧化过程中的显著差异代谢产物并预测解离常数(pKa)；结合转录组学对比分析Y-9氨氧化过程中的显著差异调控基因。【结果】Y-9在初始pH为7.19的相对酸性条件下，产生麦芽糖醇提高胞外pH；通过上调脱氨酶、脱亚胺酶和阳离子转运相关基因在相对酸性环境中的表达来维持细胞内pH稳定性。在初始pH为9.40的碱性条件下，产5-氨基戊酸3和草氨酸等有机酸及酸性物质降低胞外pH；通过调控NADH脱氢酶、细胞色素、ATP合酶和氨基酸转运相关基因的表达来维持细胞内酸度，应对碱性环境。【结论】本研究结果首先发现了Y-9具有稳定胞外pH的能力，探讨了其胞内pH稳态机制，拓展了对微生物与环境相互作用的认知，为进一步认识微生物脱氮过程中系统pH稳定机理提供了理论依据。     

Repertoire of complement in immunological defense mechanisms of fish

Fish have a serum protein system comprising a large number of complement components whose characterization is incomplete. Fish complement reveals extensive biological activities and unique properties different from mammalian complement. The first section of this review includes a concise explanation of the human complement system as a background to the understanding of the general principles of complement. The classical pathway, alternative pathway, and opsonization of the complement system are also explained. Successively, properties of fish complement are described in relation to heat lability, divalent cation requirements, species specificity, and factors affecting complement activity. Methods for the determination of complement activities involved in antibody specific and nonspecific hemolysis are also explained in detail. The titration of hemolytic antibody and its kinetics in the antibody production of salmonid fish, as an example for complement fixation, are reviewed as practical uses for fish complement. Antimicrobial activities, including bactericidal action, detoxification, viral inactivation, and opsonization in phagocytosis are also reviewed to relate complement to the defense mechanisms of fish. With respect to the bactericidal action, the modulation of complement activity in response to physiological and pathological changes, due to infections with pathogenic bacteria, was stressed as a parameter of health assessment in fish. In the last section, the ontogenetic development of salmonid complement, and interrelations between phylogenetic taxonomy and fish complement based on hemolytic activities are discussed. Such miscellaneous properties of complement activity in fish are categorized into two actions: (a) a lytic action representing hemolysis, bacteria killing, viral inactivation, etc. by the activation of the complement; and (b) and opsonic action by a fragment liberated from activated complement components.     

Effect of diesel fuel pollution on the lipid composition of some wide-spread Black Sea algae and invertebrates

Two green algae (Ulva rigida and Cladophora coelothrix), the mussel Mytilus galloprovincialis and the snail Rapana thomasiana from the Bulgarian Black Sea shore have been treated with diesel fuel (100mg l(-1)) in an aquarium with sea-water for three days. The lipids and their fatty acid changes have been examined. Significant changes have been observed mainly in the polar lipids and in the saturation of the fatty acids. These changes appeared to be bigger in the evolutionary less advanced species from both groups of marine organisms--algae and invertebrates (Ulva rigida and Mytilus galloprovincialis respectively). The data obtained could be used for a biomonitoring of the pollution.     

Review on the effects of pollution on marine fish life and fisheries in the North Sea

Information on possible effects of pollution on fish life is reviewed with special emphasis to the North Sea. Concentrations of heavy metals are high in onshore waters, sediments and organisms. Offshore regions are considered to be not endangered. In estuaries concentrations of certain heavy metals are often surpassing thresholds, which have been experimentally demonstrated to produce sublethal effects on marine organisms. Also mussel culture areas had to be closed due to excess of cadmium contamination. Mercury in fillets of cod in the southern North Sea sometimes reaches 0.3 ppm representing 1/3 of the legal limit. Reductions of inputs are necessary due to the bioaccumulative and toxic potential of heavy metals. Organochlorine contamination of water, sediments and organisms is higher in the southern than in the northern North Sea. In offshore areas clearly elevated levels of PCB's, which are the dominating organochlorine compound in marine organisms, have been analyzed. At present only a minor fraction of total organochlorine residues accumulated in marine biota is analyzed. Margins between experimentally derived lethal levels and those occurring in marine biota are narrow suggesting that sublethal effects might occur. One of the special concerns in the context of organochlorine contamination has to be seen in the potential of these substances affecting the reproductive capacity of marine organisms. Reduction measures are urgently necessary and should include substances with bioaccumulative and toxicological potential, which are not presently included in Marine Monitoring Programmes. Impact of oil pollution is considered to be a regional problem, although especially in estuaries concentrations in the water column might be high. Input through oil platforms has led to local reductions in benthic communities. Measures have to be taken, to further reduce these inputs also in view to avoid contamination of fish in the vicinity of oil fields. Oxygen depletion in German and Danish coastal waters has been shown to be a severe degradation in coastal water quality resulting in significant impact on fish populations. Indications for large-scale avoidance reactions of low DO-areas and mortalities of fish and benthos organisms have been demonstrated. It is suspected that the low DO-situations are related to increased nutrients input and high phytoplankton production. Improvement of the situation can only be expected, if input of nutrients is reduced. Reasons for the occurrence of toxic plankton blooms are not understood. An intensified research is necessary, to avoid damage to fisheries and mariculture. Although activity of radionucleids in North Sea waters is increased due to input of radioactive material from La Hague and Sellafield reprocessing units, the present levels of activity are not considered to be a threat for marine life. The same holds for thermal pollution, which is, depending on the recipient area, expected to have only local effects. Sewage sludge dumping and discharge of untreated sewage introduce significant amounts of accompanying heavy metals, which can be expected to have a negative influence on the total metal budget of recipient waters. Nutrient input through sewage will exhibit only local effects. Dumping of wastes from titanium dioxide industry are suspected to be causally linked to the occurrence of increased prevalences of certain diseases of dab in the German Bight. Present management strategies should take account of the fact that through dumping of waste from titanium dioxide production significant quantities of heavy metal are introduced into the southern North Sea. Also the incineration of chemical wastes at sea should be reduced due to uncertainties concerning quantities and toxicity of incineration products and their potential impact on marine life. It is concluded that clear evidence exists for pollution impact in estuarine regions. Most of the major estuaries at the North Sea coast show benthos reductions, which can be related to water quality and it has to be assumed that areas of a sublethal effect are located in the vicinity of those areas of well demonstrated effects. They are of unknown size and attempts have to be undertaken to quantify areas of sublethal impact. The occurrence of fish diseases is interpreted as an expression of degraded water quality. Some of the diseases detected can produce mortality, thereby having an as yet unquantified impact on fish populations. The presence of fish diseases also reduces the marketability of catches. It cannot be excluded that changes in species composition of fish populations in German Wadden Sea areas are related to pollution. Concentrations of organochlorine substances in fishes of the southern North Sea give rise to the concern that their reproductive capability might be impaired. Future work has to be directed to this problem. Impact of pollution on populations of marine organisms has so far not been demonstrated.     

Influence of anthropogenic pollution on ostracod assemblages of Amurskii Bay,Sea of Japan

The species composition and distribution of ostracods were investigated at two sites with different pollution levels on the eastern coast of Amurskii Bay within the limits of Vladivostok City. A total of 41 species were found. In all, thirty-eight species (28 of them alive) were found at the first site between Krasnyi and Groznyi capes. Another twenty-seven species were found at a depth of 1.5–3 m in the phytal zone with a diversity of microbiotopes. As the bottom became increasingly silty and the depth increased, the number of species decreased. A total of 25 species (only 15 of them alive) were found at the mouth of the Vtoraya Rechka River, which is heavily polluted by municipal and industrial sewage discharge. No valves of ostracods were found in surface sediments on silts at a depth down to 5.5m. At 4 m, only 2 species were found alive on stones overgrown with Saccharina japonica. At 500 m from a sewage discharge site, few ostracod valves were found that seemed to have died recently. At a distance of over 1 km, an ostracod assemblage typical of the silty substrates of Amurskii Bay was found below a 7 m depth (21 species, 12 of them alive).     

White Blood Cell Count of the scorpion fish <Emphasis Type="Italic">Scorpaena porcus</Emphasis> as a biomarker of anthropogenic pollution in the Black Sea coastal waters

This paper outlines the research findings on alterations in White Blood Cell Count of the scorpion fish Scorpaena porcus that inhabits the bays along the coastline of Sevastopol, depending on the level of environmental pollution. In the most polluted areas, the scorpion fish exhibits lymphocytosis and neutropenia.     

Assessment of the state of ovaries of some fish species from Peter the Great Bay of the Sea of Japan in relation to its anthropogenic pollution

Ovaries of four flatfish species—Liopsetta pinnifasciata, Pleuronectes obscurus, Pleuronectes yokohamae, and Glyptocephalus stelleri, as well as of Far Eastern navaga Eleginus gracilis and rudd Tribolodon brandtii caught in Peter the Great Bay of the Sea of Japan in prespawning and/or spawning periods were studied. Under conditions of anthropogenic pollution of the bay, the state of fish ovaries is determined by several factors, such as migration activity, mode of life (bottom or pelagic), and the degree of sites pollution where females stayed in the period of the vitellogenous growth of oocytes. The pollution of the bay exerts the most damaging effect on the reproductive system of shallow species of flatfish—L. pinnifasciata and P. obscurus—that do not perform far migrations and throughout their life stay in the zone exposed to toxicants. In navaga Eleginus gracilis and flatfish P. yokohamae, and G. stelleri that perform seasonal migrations, no damages of ovaries were found.     

Measurement and mapping of the GSM-based electromagnetic pollution in the Black Sea region of Turkey

Electromagnetic pollution caused by mobile communication devices, a new form of environmental pollution, has been one of the most concerning problems to date. Consequences of long-term exposure to the electromagnetic radiation caused by cell phone towers are still unknown and can potentially be a new health hazard. It is important to measure, analyze and map the electromagnetic radiation levels periodically because of the potential risks. The electromagnetic pollution maps can be used for the detection of diseases caused by the radiation. With the help of the radiation maps of different regions, comparative analysis can be provided and distribution of the diseases can be investigated. In this article, Global System for Mobile communication (GSM)-based electromagnetic pollution map of the Rize Providence, which has high cancer rates because of the Chernobyl nuclear explosion, is generated. First, locations of the GSM base stations are identified and according to the antenna types of the base stations, safety distances are determined. Subsequently, 155 measurements are taken during November 2014 from the nearest living quarters of the Rize city center in Turkey. The measurements are then assessed statistically. Thenceforth, for visual judgment of the determined statistics, collected measurements are presented on the map. It is observed that national limits are not exceeded, but it is also discovered that the safety distance is waived at some of the measurement points and above the average radiation levels are noted. Even if the national limits are not exceeded, the long-term effects of the exposition to the electromagnetic radiation can cause serious health problems.     

Dependence of alkaline phosphatase activity on bioecological parameters of Black Sea fish

The data on alkaline phosphatase activity in the liver of eight dominant species of Black Sea fishes are presented. The enzyme activity varies within a wide range; significant sexual differences have not been registered. The activity of the enzyme is similar in specimens of different age groups, but it decreases in old fishes. The enzyme activity increases in prespawning and spawning, which evidences for its participation in the process of sexual maturation of fish. Based on our and published data on the dependence of alkaline phosphatase activity on the degree of the environmental pollution, the enzyme can be recommended as a biomarker for bioindication, biotesting, and ichthyological monitoring.     

Marine molluscs and fish as biomarkers of pollution stress in littoral regions of the Red Sea, Mediterranean Sea and North Sea

The intensive development of industry and urban structures along the seashores of the world, as well as the immense increase in marine transportation and other activities, has resulted in the deposition of thousands of new chemicals and organic compounds, endangering the existence of organisms and ecosystems. The conventional single biomarker methods used in ecological assessment studies cannot provide an adequate base for environmental health assessment, management and sustainability planning. The present study uses a set of novel biochemical, physiological, cytogenetic and morphological methods to characterize the state of health of selected molluscs and fish along the shores of the German North Sea, as well as the Israeli Mediterranean and Red Sea. The methods include measurement of activity of multixenobiotic resistance-mediated transporter (MXRtr) and the system of active transport of organic anions (SATOA) as indicators of antixenobiotic defence; glutathione S-transferase (GST) activity as an indicator of biotransformation of xenobiotics; DNA unwinding as a marker of genotoxicity; micronucleus test for clastogenicity; levels of phagocytosis for immunotoxicity; cholinesterase (ChE) activity and level of catecholamines as indicators of neurotoxicity; permeability of external epithelia to anionic hydrophilic probe, intralysosomal accumulation of cationic amphiphilic probe and activity of non-specific esterases as indicators of cell/tissue viability. Complete histopathological examination was used for diagnostics of environmental pathology. The obtained data show that the activity of the defensive pumps, MXRtr and SATOA in the studied organisms was significantly higher in the surface epithelia of molluscs from a polluted site than that of the same species from control, unpolluted stations, providing clear evidence of response to stress. Enhanced frequency of DNA lesions (alkaline and acidic DNA unwinding) and micronucleus-containing cells was significantly higher in samples from polluted sites in comparison to those from the clean sites that exhibited genotoxic and clastogenic activity of the pollutants. In all the studied molluscs a negative correlation was found between the MXRtr levels of activity and the frequency of micronucleus-containing hemocytes. The expression of this was in accordance with the level of pollution. The complete histopathological examination demonstrates significantly higher frequencies of pathological alterations in organs of animals from polluted sites. A strong negative correlation was found between the frequency of these alterations and MXRtr activity in the same specimens. In addition to these parameters, a decrease in the viability was noted in molluscs from the polluted sites, but ChE activities remained similar at most sites. The methods applied in our study unmasked numerous early cryptic responses and negative alterations of health in populations of marine biota sampled from the polluted sites. This demonstrates that genotoxic, clastogenic and pathogenic xenobiotics are present and act in the studied sites and this knowledge can provide a reliable base for consideration for sustainable development. Received: 2 March 1999 / Received in revised form: 2 August 1999 / Accepted: 3 August 1999     

Peroxisome proliferator-activated receptor gamma is essential for stress adaptation by maintaining lipid homeostasis in female fish

Reduction of lipid synthesis often causes free fatty acid (FFA) overload, resulting consequential oxidative stress and health damage. Environmental stresses also induce cellular oxidative stress in organisms. The functional peroxisome proliferator-activated receptor gamma (pparg) gene is essential for lipid synthesis and homeostatic lipid maintenance. However, the relationship between the pparg-mediated lipid synthesis and environmental stress adaptation awaits full elucidation. Here, we generated a pparg-knockout zebrafish model. The conversion of free fatty acids into triglycerides in the female pparg mutants was hampered by reduced esterification efficiency, thus induced lipotoxicity, as evidenced by high oxidative stress and damaged health in these mutants, which led to reduced resistance to cold, heat and ammonia nitrogen stresses. Activating pparg in the wild-type female fish via dietary supplementation with rosiglitazone (a pparg agonist), or reducing oxidative stress in the female pparg mutants via dietary supplementation with N-acetylcysteine (an antioxidant), or promoting mitochondrial fatty acid β-oxidation in the female pparg mutants via dietary supplementation with l-carnitine, resulted in significantly reduced cellular injury, and improved environmental stress resistance. Collectively, our findings reveal that the regulative function of pparg in FFA esterification is important in stress resistance in female fish, and highlight the tight correlation existing between lipotoxicity and environmental adaptation.     

Overfishing of top predators eroded the resilience of the Black Sea system regardless of the climate and anthropogenic conditions

It is well known that human activities, such as harvesting, have had major direct effects on marine ecosystems. However, it is far less acknowledged that human activities in the surroundings might have important effects on marine systems. There is growing evidence suggesting that major reorganization (i.e., a regime shift) is a common feature in the temporal evolution of a marine system. Here we show, and quantify, the interaction of human activities (nutrient upload) with a favourable climate (run‐off) and its contribution to the eutrophication of the Black Sea in the 1980s. Based on virtual analysis of the bottom‐up (eutrophication) vs. top‐down (trophic cascades) effects, we found that an earlier onset of eutrophication could have counteracted the restructuring of the trophic regulation at the base of the food web that resulted from the depletion of top predators in the 1970s. These enhanced bottom‐up effects would, however, not propagate upwards in the food web beyond the zooplankton level. Our simulations identified the removal of apex predators as a key element in terms of loss of resilience that inevitably leads to a reorganization. Once the food web has been truncated, the type and magnitude of interventions on the group replacing the apex predator as the new upper trophic level have no effect in preventing the trophic cascade. By characterizing the tipping point at which increased bottom‐up forcing exactly counteracts the top‐down cascading effects, our results emphasize the importance of a comprehensive analysis that take into account all structuring forces at play (including those beyond the marine system) at a given time.     

Regulation of immunological homeostasis in the respiratory tract

The respiratory tract has an approximate surface area of 70 m2 in adult humans, which is in virtually direct contact with the outside environment. It contains a uniquely rich vascular bed containing a large pool of marginated T cells, and harbours a layer of single-cell-thick epithelial tissue through which re-oxygenation of blood must occur uninterrupted for survival. It is therefore not surprising that the respiratory tract is never more than a short step away from disaster. We have only a partial understanding of how immunological homeostasis is maintained in these tissues, but it is becoming clear that the immune system has evolved a range of specific mechanisms to deal with the unique problems encountered in this specialized microenvironment.     

Length–weight relationships for seven fish species from the southeastern Black Sea

This study reports the parameters a and b of the length–weight relationships for seven fish species (Gobius niger, Mesogobius batrachocephalus, Mullus barbatus, Neogobius melanostomus, Raja clavata, Scorpaena porcus, Uranoscopus scaber) from the southeastern Black Sea.     

Peculiarities of the phospholipid and fatty acid composition of erythrocyte plasma membranes of the Black Sea fish

The phospholipid and the fatty acid composition of the main phospholipids families of erythrocyte plasma membranes was studied in two species of cartilaginous fish: the common thrasher (Raja clavata L.) and the common stingray (Dasyatis pastinaca) and three bony fish species: the scorpion fish (Scorpaena porcus L.), the smarida (Spicara flexuosa Raf.), and the horse mackerel (Trachurus mediterraneus ponticus Aleev). It was shown that in the studied fish, 70.0-80.0 % of all membrane phospholipids were composed of phosphatidylcholine and phosphatidylethanolamine. Phosphatidylserine, monophosphoinositide, and sphingomyelin were minor components whose content in the erythrocyte membrane fluctuated from 3.0 % to 13.0 %. The fatty acid phospholipids composition was represented by a large specter of acids. From saturated acids, basic for plasma membranes are palmitic (C16: 0) and stearic (C18: 0) acids. From unsaturated acids, the larger part belong to mono-, tetra-, penta-, and hexaenoic acids in fish phospholipids. The calculation of the double bond index and of the unsaturation coefficient showed difference in the deformation ability of erythrocyte membranes of the studied fish.     

Findings of new and rare fish species in the coastal zone of the Crimea (the Black Sea)

Characteristics of seven Atlantic-Mediterranean fish species are provided. Of them, four (gobies—yellow-headed Gobius xanthocephalus and red-mouthed G. cruentatus, incognito blenny Parablennius incognitus, and greater pipefish Syngnathus acus) were either first found in the Black Sea coastal zone of the Crimea, or their findings were doubted, and three species (thin-lipped grey mullet Liza ramada, European barracuda Sphyraena sphyraena, and Black Sea salmon Salmo trutta labrax) are very rare. Over the last decade, the authors recorded 15 fish species; of them, seven Atlantic-Mediterranean species have dispersed along the Crimean coasts by now, one endemic of the far eastern seas (chameleon goby Tridentiger trigonocephalus) has naturalized in Sevastopol Bay, four invading species, freshwater by origin, permanently occur in the estuary zones of the peninsula; the remaining three species are known from single findings and apparently belong to the category of accidental fish. An increase in the species composition of ichthyofauna off the Black Sea coasts of the Crimea results from the on-going process of mediterranization—the invasion in the estuary of freshwater tolerant species at the expense of accidental species that penetrated here either independently or with ballast waters.