A Review of Nontraditional Biomanipulation

The aim of this review is to identify problems, find general patterns, and extract recommendations for successful management using nontraditional biomanipulation to improve water quality. There are many obstacles that prevent traditional biomanipulation from achieving expectations: expending largely to remove planktivorous fish, reduction of external and internal phosphorus, and macrophyte re-establishment. Grazing pressure from large zooplankton is decoupled in hypereutrophic waters where cyanobacterial blooms flourish. The original idea of biomanipulation (increased zooplankton grazing rate as a tool for controlling nuisance algae) is not the only means of controlling nuisance algae via biotic manipulations. Stocking phytoplanktivorous fish may be considered to be a nontraditional method; however, it can be an effective management tool to control nuisance algal blooms in tropical lakes that are highly productive and unmanageable to reduce nutrient concentrations to low levels. Although small enclosures increase spatial overlap between predators and prey, leading to overestimates of the impact of predation, microcosm and whole-lake experiments have revealed similar community responses to major factors that regulate lake communities, such as nutrients and planktivorous fish. Both enclosure experiments and large-scale observations revealed that the initial phytoplankton community composition greatly impacted the success of biomanipulation. Long-term observations in Lake Donghu and Lake Qiandaohu have documented that silver carp (Hypophthalmichthys molitrix) and bighead carp (H. nobilis) (two filter-feeding planktivorous species commonly used in management) can suppress Microcystis blooms efficiently. The introduction of silver and bighead carp could be an effective management technique in eutrophic systems that lack macrozooplankton. We confirmed that nontraditional biomanipulation is only appropriate if the primary aim is to reduce nuisance blooms of large algal species, which cannot be controlled effectively by large herbivorous zooplankton. Alternatively, this type of biomanipulation did not work efficiently in less eutrophic systems where nanophytoplankton dominated.

Kinetic study of the 2-methyl-3-methoxy-4-phenylbutanoic acid produced by oxidation of microcystin in aqueous solutions

Microcystins (MCs) are a family of related cyclic hepatotoxic heptapeptides, of which more than 70 types have been identified. The chemically unique nature of the C20 beta-amino acid, (2S, 3S, 8S, 9S)-3-amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca4,6-dienoic acid (Adda), portion of the MCs has been exploited to develop a strategy to analyze the entirety. Oxidation of MCs causes the cleavage of MC Adda to form 2-methyl-3-methoxy-4-phenylbutanoic acid (MMPB). In the present study, we investigated the kinetics of MMPB produced by oxidation of the most-often-studied MC variant, MC-LR (L = leucine, R = arginine), with permanganate-periodate. This investigation allowed insight regarding the influence of the reaction conditions (concentration of the reactants, temperature, and pH) on the conversion rate. The results indicated that the reaction was second order overall and first order with respect to both permanganate and MC-LR. The second-order rate constant ranged from 0.66 to 1.35 M/s at temperatures from 10 to 30 degrees C, and the activation energy was 24.44 kJ/mol. The rates of MMPB production can be accelerated through increasing reaction temperature and oxidant concentration, and sufficient periodate is necessary for the formation of MMPB. The initial reaction rate under alkaline and neutral conditions is higher than that under acidic conditions, but the former decreases faster than the latter except under weakly acidic conditions. These results provided new insight concerning selection of the permanganate-periodate concentration, pH, and temperature needed for the oxidation of MCs with a high and stable yield of MMPB.

Accumulation of hepatotoxic microcystins in freshwater mussels, aquatic insect larvae and oligochaetes in a large, shallow eutrophic lake (Lake Chaohu) of subtropical China

This paper studied the seasonal changes of two common microcystins (MCs), MC-RR and -LR, in the commercially important mussel Corbicula fluminea in Lake Chaohu, where there occurred dense cyanobacteria. Occasional measurements were also made for MC in the mussel Arconaia lanceolat, the oligochaete Limnodilus hoffineisteri and the insect larva Chironomus sp. Mean MC of C. fluminea was much higher in hepatopancreas than in intestine and foot. Our study is the first to report accumulation of MCs in oligochaetes and aquatic insect larvae. The hi-h contents of MCs in the insect larvae suggest a great possibility for the transfer of MCs to benthos-feeding omnivores like common carp. According to the provisional standard by the WHO, 28.6% of the collected C. fluminea were harmful for human consumption, assuming a daily consumption of 300 by a person. It is recommended that edible mussels should not be collected for human consumption during toxic cyanobacterial blooms in Lake Chaohu.

In situ study on effect of food competition on diet shifts and growth of silver and bighead carps in large biomanipulation fish pens in Meiliang Bay, Lake Taihu

Silver and bighead carps were cultured in large fish pens to reduce the risks of cyanobacterial bloom outbreaks in Meiliang Bay, Lake Tauhu in 2004 and 2005. Diet compositions and growth rates of the carps were studied from April to November each year. Both carp species fed mainly on zooplankton (> 50% in diet) in 2004 when competition was low, but selected more phytoplankton in 2005 when competition was high. Silver carp had a broader diet breadth than did bighead carp. Higher densities and fewer food resources increased diet breadths but decreased the diet overlap in both types of carps. It can be predicted that silver and bighead carps would be released from diet competition and shift to feed mainly on zooplankton at low densities, decreasing the efficiency of controlling cyanobacterial blooms. Conclusively, when silver and bighead carps are used to control cyanobacterial blooms, a sufficiently high stocking density is very important for a successful practice.

Field studies on the environmental factors in controlling microcystin production in the subtropical shallow lakes of the Yangtze River

Microcystin (MC) problem made more and more care about in China, intercellular MC (Int-MC) and cellular MC (Cel-MC) were important contents to reflect the producing-MC ability by cyanobacteria and by lakes. To study the correlations between Int-MC, Cel-MC concentration and biological and environmental factors, eight cyanobacterial blooming lakes were studied in the middle and lower reaches of the Yangtze River. Microcystin-RR (MC-RR) and Microcystin-LR (MC-LR) were the primary toxin variants in our data. From the linear correlations between MC and environmental factors, cellular-YR had significant correlation with most of chemical factors except total nitrogen (TN) and the ratio of total nitrogen and total phosphorus (TN/TP), most intracellular MC analogues had significant correlations with total dissolved nitrogen (TDN), ammonium (NH4+), nitrite (NO2-), TP, total dissolved phosphorus (TDP), Microcystis. From the canonal correspondence analysis, Int-MC concentrations were closely related with the chemical and biological factors, such as TP, total organic carbon (TOC), chlorophyll a (Chl a), Microcystis biomass, et al. While Cel-MC contents, especially Cel-RR and Cel-LR, were closely related with light environmental in the lakes such as water depth and transparence.

Field and laboratory studies on pathological and biochemical characterization of microcystin-induced liver and kidney damage in the phytoplanktivorous bighead carp

Field and experimental studies were conducted to investigate pathological characterizations and biochemical responses in the liver and kidney of the phytoplanktivorous bighead carp after intraperitoneal (i.p.) administration of microcystins (MCs) and exposure to natural cyanobacterial blooms in Meiliang Bay, Lake Taihu. Bighead carp in field and laboratory studies showed a progressive recovery of structure and function in terms of histological, cellular, and biochemical features. In laboratory study, when fish were i.p. injected with extracted MCs at the doses of 200 and 500 mu g MC- LReq/kg body weight, respectively, liver pathology in bighead carp was observed in a time dose-dependent manner within 24 h postinjection and characterized by disruption of liver structure, condensed cytoplasm, and the appearance of massive hepatocytes with karyopyknosis, karyorrhexis, and karyolysis. In comparison with previous studies on other fish, bighead carp in field study endured higher MC doses and longer-term exposure, but displayed less damage in the liver and kidney. Ultrastructural examination in the liver revealed the presence of lysosome proliferation, suggesting that bighead carp might eliminate or lessen cell damage caused by MCs through lysosome activation. Biochemically, sensitive responses in the antioxidant enzymes and higher basal glutathione concentrations might be responsible for their powerful resistance to MCs, suggesting that bighead carp can be used as biomanipulation fish to counteract cyanotoxin contamination.

Dose-dependent effects of extracted microcystins on embryonic development, larval growth and histopathological changes of southern catfish (Silurus meridionalis)

A laboratory toxic experiment was conducted to examine dose-dependent effects of extracted microcystins (MCs) on embryonic development, larval growth and histopathological changes of southern catfish (Silurus meridionalis). Fertilized eggs were incubated in solutions with four concentrations of MCs (0, 1, 10, 100 mu g MC-LReq l(-1)). Higher MCs retarded egg development (2-10 h delays) and larval growth, reduced hatching rate (up to 45%), and caused high malformation rate (up to 15%) and hepatocytes damage (characterized by disorganization of cell structure and a loss of adherence between hepatocytes, cellular degeneration with vacuolar hepatocytes and marginal nuclei, even hepatocellular necrosis). A 10 mu g MC-LReql(-1) is close to a high concentration in natural cyanobacterial blooms, suggesting a possible existence of such toxic effects in eutrophic waters. (c) 2007 Elsevier Ltd. All rights reserved.

Reduction in microcystin concentrations in large and shallow lakes: Water and sediment-interface contributions

Blooms of cyanobacteria, or blue-greens, are known to produce chemicals, such as microcystins, which can be toxic to aquatic and terrestrial organisms. Although previous studies have examined the fate of microcystins in freshwater lakes, primary elimination pathways and factors affecting degradation and loss have not been fully explained. The goal of the present study was to explore sources of algal toxins and investigate the distribution and biodegradation of microcystins in water and sediment through laboratory and field analyses. Water and sediment samples were collected monthly from several locations in Lake Taihu from February 2005 to January 2006. Samples were analyzed for the presence of microcystin. Water and sediment were also used in laboratory studies to determine microcystin degradation rates by spiking environmental samples with known concentrations of the chemical and observing concentration changes over time. Some water samples were found to efficiently degrade microcystins. Microcystin concentrations dropped faster in water collected immediately above lake sediment (overlying water). Degradation in sediments was higher than in water. Based on spatial distribution analyses of microcystin in Lake Taihu, higher concentrations (relative to water concentrations) of the chemical were found in lake sediments. These data suggest that sediments play a critical role in microcystin degradation in aquatic systems. The relatively low levels of microcystins found in the environment are most likely due to bacterial biodegradation. Sediments play a crucial role as a source (to the water column) of bio-degrading bacteria and as a carbon-rich environment for bacteria to proliferate and metabolize microcystin and other biogenic toxins produced by cyanobacteria. These, and other, data provide important information that may be applied to management strategies for improvement of water quality in lakes, reservoirs and other water bodies. (C) 2007 Elsevier Ltd. All rights reserved.

High variability of the gvpA-gvpC region in Microcystis

Gas vesicles provide buoyancy to Microcystis and other common cyanobacterial bloom-forming species. gvpA and gvpC are structural genes encoding gas vesicle proteins. Phylogenetic analyses of 10 Microcystis strains/uncultured samples showed that gvpC and each intergenic segment of the gvpA-gvpC region can be divided into two types. The combination of different types of gvpC and intergenic segments is an important factor that diversifies this genomic region. Some Microcystis strains isolated in China possess a 172 to 176 bp sequence tag in the intergenic segment between gvpA and gvpC. The gvpA-gvpC region in Microcystis can be divided into at least 4 classes and more numbers of subclasses. Compared to rbcLX and other regions, the high variability of the gvpA-gvpC region should be more useful in identifying geographical isolates or ecotypes of Microcystis.

Effects of microcystin-RR on the antioxidant system of Bacillus subtilis

Microcystins are a kind of cyclic hepatotoxins produced by many cyanobacterial species. Many works have been done concerning, the toxic effects of microcystins on animals and plants. However, the reports about their effects on microbial cells are very limited. In the present paper, Bacillus subtilis (B. subtilis) was used to determine the dose- and time-effect of microcystin-RR, and the results showed that the activity of antioxidant enzymes including superoxide dismutase (SOD) and catalase (CAT) was significantly increased to that of control, when exposed to 5 or 10 mu g/ml microcystin-RR for 1 h. The contents of thiobarbituric acid-reactive sub-stances (TBARS) and glutathione (GSH) as well as glu-tathione reductase (GR) activity were obviously increased only when exposed to 10 mu g/ml microcystin-RR. For the time-effect of microcystin-RR on B. subtilis, the activities of antioxidant enzymes including SOD and CAT as well as GR activity and TBARS, GSH contents in B. subtilis were at first significantly increased, and then subsequently de-creased. These results suggested that microcystin-RR could induce the oxidative stress of B. subtilis for a short period. The antioxidant system protects B. subtilis from oxidative damage.

Allelopathic interactions between Potamogelon maackianus and Microcystis aeruginosa

The allelopathic interactions between Potamogeton maackianus and toxic cyanobacteria (Microcystis aeruginosa) were studied. P maackianus inhibited the growth of M. aeruginosa, both in a coexistence culture system and in exudates experimental culture system. M. aeruginosa also showed effects on the secondary metabolic biosynthesis and secreting behavior of P maackianus. The main lipophilic components of the hexane extracts and the exudates from the macrophyte were analyzed through GC-MS determination. The lipophilic components of the hexane extracts and the exudates from P. maackianus were influenced by M. aeruginosa or their released chemicals. Comparing the lipophilic constituents of the hexane extracts with those in the exudates, the results showed that weak polar compounds contained in the macrophytes can be secreted into the surrounding water. Many compounds, such as N-phenyl-2-naphthalenamine and isopropyl myristate, were detected both in the hexane extracts and the exudates. The changes of lipophilic components in the hexane extracts would be a response to the stress of toxic cyanobacteria or their released toxins. Those changes of exudates, especially the increased content of N-phenyl-2-naphthalenamine, might also be an induced defensive behavior mediated by the released toxins from M aeruginosa. The study results about reciprocal allelopathic responses between macrophytes and cyanobacteria can help in the management of eutrophic waters, and is also important information concerning strategies for recovering eutrophic waters.

Phytoplankton biomass in relation to nutrients and zooplankton in thirty subtropical lakes adjacent to the Yangtze River, China

A total of 30 shallow lakes, located along the middle and lower reaches of the Yangtze River, were studied to assess the relative importance of nutrients and zooplankton biomass in determining the phytoplankton biomass in subtropical China. Zooplankton biomass and nutrients both varied greatly in these lakes. Factor analysis and multiple linear regression showed that phytoplankton biomass was positively correlated with TN, NH4+, NO3- and TP, while it did not show any negative relationship to zooplankton biomass. Meanwhile, the phytoplankton biomass showed contrary relationships to the mass ratio of TN/TP in spring and summer, suggesting that in nutrient-richer lakes the dominant phytoplankton species have different preferences for TN/TP ratio. The insignificant top-down control of phytoplankton biomass may be attributed to the dominance of small-sized crustaceans and low crustacean biomass resulting from cyanobacterial dominance and planktivorous fish predation as well as other factors. Thus, it is likely that nutrients were more important than zooplankton biomass in explaining the total variance of phytoplaDkton biomass in these subtropical lakes.

Identification of a gene, ccr-1 (sll1242), required for chill-light tolerance and growth at 15 degrees C in Synechocystis sp PCC 6803

Synechocystis sp. PCC 6803 exposed to chill (5 degrees C)-light (100 mu mol photons m(-2) s(-1)) stress loses its ability to reinitiate growth. From a random insertion mutant library of Synechocystis sp. PCC 6803, a sll1242 mutant showing increased sensitivity to chill plus light was isolated. Mutant reconstruction and complementation with the wild-type gene confirmed the role of sll1242 in maintaining chill-light tolerance. At 15 degrees C, the autotrophic and mixotrophic growth of the mutant were both inhibited, paralleled by decreased photosynthetic activity. The expression of sll1242 was upregulated in Synechocystis sp. PCC 6803 after transfer from 30 to 15 degrees C at a photosynthetic photon flux density of 30 mu mol photons m(-2) S-1. sll1242, named ccr (cyanobacterial cold resistance gene)- 1, may be required for cold acclimation of cyanobacteria in light.

Microcystin accumulation in freshwater bivalves from Lake Taihu, China, and the potential risk to human consumption

The potential risk through ingestion of microcystins (MC) in contaminated mollusks has not been well studied. The present paper studied seasonal changes of MC content (determined by liquid chromatography-mass spectrometry) in various organs of three species of bivalves (Cristaria plicata, Hyriopsis cumingii, and Lamprotula leai) in Lake Taihu, China, where toxic cyanobacterial blooms occurred. Coinciding with peaks of seston MC (maximum, 5.7 mu g/L) and MC in cyanobacterial blooms (maximum, 0.534 mg/g), most organs showed sharp MC peaks during the summer, indicating both fast uptake and fast depuration by bivalves. Because hepatopancreas and intestine had considerably higher MC content than other organs, they are the most dangerous for human consumption. Both the present and previous studies show that the hepatopancreatic MC and total tissue MC often are correlated in various aquatic invertebrates. During the peak of the cyanobacterial blooms, C. plicata had higher hepatopancreatic MC content than the other bivalves, whereas H. cumingii had higher intestinal MC content than the other bivalves. Estimated daily intakes for humans from the consumption of whole tissues of the three bivalves were 0.48 to 0.94 mu g MC-LR equivalent/kg body weight (12- to 23.5-fold the tolerable daily intake value proposed by the World Health Organization), which indicates a high risk for humans consuming these bivalves.