Can algicide (the thiazolidinedione derivative TD49) truly contribute to the restoration of microbial communities?

Harmful algal blooms (HABs) are becoming a more serious ecological threat to marine environments; they not only produce toxins, resulting in the death of marine organisms, but they also adversely affect biodiversity, which is an indicator of the health of an ecosystem. Thus, to mitigate HABs, numerous studies have been conducted to develop an effective algicide, but few studies have elucidated the effect of algicides on marine environmental health. In this study, thiazolidinedione derivative 49 (TD49), which has been developed as an algicide for the dinoflagellate Heterocapsa circularisquama, was used, and we investigated changes in phytoplankton biomass (abundance, chlorophyll a, and carbon biomass) and biodiversity (diversity, evenness, and richness) following the application of TD49. To gain deeper understanding, a large-scale mesocosm (1300 L) experiment containing control and treatment with four different concentrations (0.2, 0.4, 0.6 and 1 µM) was conducted for 10 days. Based on a previous study, TD49 shows algicidal activity against H. circularisquama depending on its concentration. The phytoplankton biomass in the TD49 treatments was generally lower than that in the control due to the algicidal effect of TD49 on H. circularisquama. The biodiversity indices (e.g., the Shannon-Weaver index) in the treatments were consistently higher than those in the control before depletion of nitrite + nitrate. Interestingly, the 0.6 µM TD49 treatment had higher biodiversity indices than the high-concentration treatment (1 µM), which appeared to show a better algicidal effect. These findings suggest that mitigation of H. circularisquama blooms with TD49 treatment may enhance phytoplankton biodiversity, but treatment with excessively high concentrations can lead to harmful effects. During the study period, regardless of the control and TD49 treatments, the total biomass of phytoplankton gradually decreased from the midpoint of the experiment to the end of the experiment. This was more likely caused by the depletion of nutrients than by the toxicity of TD49.

Effects of the algicide, thiazolidinedione derivative TD49, on microbial communities in a mesocosm experiment.

We investigated the effects of the algicide thiazolidinedione derivative TD49 on microbial community in mesocosm experiments. The TD49 concentration exponentially decreased over time, with half-life of 3.5 h, following addition in the seawater (R2=0.98, P<0.001). Among microbial communities, heterotrophic bacteria and heterotrophic nanoflagellates (HNFs) grew well in all treatments following the addition of TD49. The abundance of HNFs lagged behind the increase in heterotrophic bacteria by 24 h in the 0.2 and 0.4 µM TD49 concentrations (R2=0.28, P<0.05), and by 48 h in the 0.6 and 1.0 µM TD49 concentrations (R2=0.30, P<0.05). This implies a strong concentration-dependent top-down effect of TD49 on microbial communities, with indications that the degradation of planktonic organisms, including the target alga, led to high heterotrophic bacteria concentrations, which in turn stimulated the population growth of predatory HNF. However, total ciliate numbers remained relatively low in the TD49 treatments relative to the control and blank groups, suggesting limited carbon flow from bacteria to these grazers even though the abundance of aloricate ciliates gradually increased toward the end of the experimental period, particularly at the high TD49 concentrations. TD49 appears to provide an environmentally safe approach to the control of harmful algal blooms (HABs) in aquatic ecosystems.

Toxic effects of Microcystis cell extracts on the reproductive system of male mice.

In this paper, the reproductive toxicity of male mice treated with Microcystis aeruginosa cell extracts containing microcystins was examined. In contrast to the control group, male mice exposed intraperitoneally to 3.33 or 6.67 microg microcystins/kg body weight for 14 days had decreased mean body weight, and the mean absolute weight of the testes and epididymides was decreased. However, the mean relative weight of the testes increased compared to the controls. In addition, histological examination of microcystin-treated mice indicated that the testes were damaged and the space between the seminiferous tubules was more pronounced compared to control mice. The quality of mature sperm in the seminiferous tubules was also decreased in treated mice compared with the control group. Further studies showed that motility and viability of the sperm from microcystin-treated mice were reduced, but no significant difference was found in the concentration and abnormality of the sperm from treated mice compared to the control. This study indicated that microcystins had numerous toxic effects on the reproductive system of male mice.

Microcystins cause embryonic toxicity in mice.

Microcystins produced by freshwater cyanobacteria are potent hepatotoxins and can cause animal intoxications and human illnesses. In the present study, the effects of microcystins on the embryonic development of Kunming mice were determined using cell extracts of Microcystis aeruginosa from the Nanwan reservoir, China. Forty-eight pregnant mice were divided into four groups of 12 mice. Pregnant mice in three experimental groups were injected intraperitoneally with cell extracts at doses equivalent to 3, 6, or 12 microg microcystins/kg body weight daily from gestational days 6-15, while the mice in the control group were injected on the same schedule with sterilized saline. Mice were killed on the 18th day of gestation and embryonic and fetal developmental indexes checked. The fetal mice were also examined for anomalies of external, skeletal, and internal organs. The results demonstrated a significant decrease in body weight gain of pregnant mice in the 12 microg/kg dose group when compared to the control group (p<0.05). Differences in mean body weight, body length, and tail length of the fetuses were also found in these two groups (p<0.05). However, no significant difference in these characteristics was detected in the 6 or 3 microg/kg dose groups when compared to the control group (p>0.05). Four fetuses in the 6 microg/kg body dose group were found to have a curving tail. Additionally, petechial hemorrhage and hydropic degeneration were observed in the livers of fetuses in the 6 and 12 microg/kg experimental groups. These results suggested that microcystins had both maternal and embryonic toxicity in mice.

Oral exposure to Microcystis increases activity-augmented antioxidant enzymes in the liver of loach (Misgurnus mizolepis) and has no effect on lipid peroxidation.

Recently, eutrophication has induced severe cyanobacterial blooms in the Naktong River, the second largest river of Korea. In the present study, lipid peroxidation and the antioxidant enzymes superoxide dismutase, catalase, and glutathione peroxidase, were evaluated in the liver of loach (Misgurnus mizolepis) that were orally exposed to a low dose of Microcystis through dietary supplementation with bloom scum. Loach received 75 mg of dry cells/kg body weight mass (equal to 10 microg microcystin-RR/kg body mass), for 28 days under controlled conditions. Antioxidant enzymatic activity and lipid peroxidation were measured after termination of exposure. The activities of antioxidant enzyme were significantly increased in the livers of toxin-exposed loach after 28 days of exposure, as compared to control fish. However, lipid peroxidation remained stable in both groups. These results suggest that antioxidant enzymes were able to eliminate oxidative stress induced by low concentrations of microcystins and to prevent increased lipid peroxidation in the liver of loach.

Subchronic oral toxicity of microcystin in common carp (Cyprinus carpio L.) exposed to Microcystis under laboratory conditions.

The subchronic oral toxicity of microcystin in common carp (Cyprinus carpio L.) was investigated in this study. The fish (mean body weight of 322+/-36 g, n=10) were orally exposed to Microcystis by feeding with bloom scum at a dose of 50 microg microcystins/kg body weight under laboratory conditions for 28 days. Growth assay results showed that microcystin could completely inhibit the growth of carp, but failed to change the fish hepatosomatic index. Ultrastructural examination by electron microscope revealed severe damage in hepatocytes derived from the treated fish. Serum biochemical assays with commercial kits indicated that alanine aminotransferase and aspartate aminotransferase activities were significantly increased as compared to control levels, but gamma-glutamyl transferase, alkaline phosphatase and lactate dehydrogenase activities remained unchanged. Protein phosphatase inhibition assay revealed that the microcystin concentrations were 261.0+/-108.3 ng microcystin-LR equivalent/g fresh weight in hepatopancreas and 38.3+/-12.3 ng microcystin-LR equivalent/g fresh weight in muscle. The latter is above the limit recommended by the World Health Organization for human consumption. Therefore, we recommend that a warning system be instituted for announcing the occurrence of microcystin-producing water bloom and the possible risk of human intoxication.