Growth inhibition in Japanese medaka (Oryzias latipes) fish exposed to tetrachloroethylene

A recent study in our laboratory has demonstrated that tetrachloroethylene (TCE) is acutely toxic to Japanese medaka (Oryzias latipes) larvae with a 96 hr-LC50 of 18 (17-19) mg/mL (Spencer et al., 2002). In the present study we hypothesize that TCE exposure induces a developmental effect in Japanese medaka. Growth and age specific sensitivity of Japanese medaka larvae were studied with four age groups (7, 14, 21 and 28 days old) to determine tetrachloroethylene effects on these parameters. The medaka larvae were exposed for 96 hours in a single concentration (10 mg/mL) of TCE. The toxic endpoints evaluated were larvae weight, length, water content and protein concentration. The study revealed that exposure of medaka larvae to this sub-acute concentration of TCE significantly reduced length and weight in the treated group. The difference in growth between control and treated groups was more obvious in age versus length, than in age versus weight. The dry weight-fresh weight ratio (dw/fw) was shown to be higher in the control group. Water content in TCE-treated medaka was higher than in the control group, and younger fry had more water content than older ones. A higher protein concentration was also observed in TCE-treated medaka compared to the control group. These results indicate that TCE has a profound effect on the growth and development of Japanese medaka larvae.     

Developmental arrest in Japanese medaka (Oryzias latipes) embryos exposed to sublethal concentrations of atrazine and arsenic trioxide

Embryos of the Japanese medaka (Oryzias latipes) were exposed to serial concentrations of atrazine (0, 25, 50, and 100 ppm) and arsenic trioxide (0, 0.025, 0.05, 0.1 ppm) until hatching. Stasis of circulation, blood islands, titanic convulsions, tube heart and mortality were observed in atrazine-treated embryos. Each endpoint exhibited a concentration-response relationship. Only 4% of the embryos hatched in the 25 ppm, and none in the 50 and 100 ppm, probably due to cell death attributed to the embryos' inability to break from the chorion. With arsenic exposure, hatching was inversely correlated to chemical concentration: 86%, 75% and 54% for 0.025, 0.05 and 0.1 ppm, respectively. Hatching periods were also reduced from 7-13 days in controls to 7-11 days in arsenic-treated embryos. This observation was more pronounced with the 0.05 ppm concentration, showing a reduction of about 4 days. Despite this shortage in hatching time, there were no observable morphological abnormalities, as seen with atrazine. The ecological significance of these findings and implications for the development of sublethal toxicity tests using Japanese medaka embryos are important.     

The effects of ammonium sulphate and urea upon egg hatching and miracidial survival of Schistosoma mansoni

The effects of various concentrations of ammonium sulphate and urea on egg hatching and miracidial survival of S. mansoni were tested in order to determine if the use of these fertilizers in the ricelands of the Republic of Cameroon could affect the transmission of schistosomiasis. Results indicate that hatching of eggs and survival of miracidia varied with concentrations of tested chemicals and times of exposure. Exposure of S. mansoni eggs to 0.20%-1.00% ammonium sulphate or to 0.50%-4.00% urea reduced their ability to hatch and produce miracidia. A chemical concentration of 1.00% ammonium sulphate or 4.00% urea was found to be sufficient to produce complete inhibition of hatching. High concentrations of both chemicals not only inhibited miracidial emergence but also may be ovocidal. Results obtained from miracidial survival tests indicated that LC5, LC50 and LC95 values for ammonium sulphate were 0.07%, 0.80% and 10.61% after 2 hours of exposure; 0.03%, 0.16% and 0.90% after 4 hours of exposure; and 0.30%, 0.20% and 0.40% after 6 hours of exposure respectively. Similar statistical analyses revealed that the LC5, LC50 and LC95 values for urea were 0.22%, 1.90% and 16.25% after 2 hours of exposure; 0.28%, 0.57% and 1.14% after 4 hours of exposure; and 0.13%, 0.27% and 0.57% after 6 hours of exposure respectively. Although the two fertilizers exerted some adverse effects on S. mansoni eggs and miracidia at relatively high concentrations, neither of them was found to be of practical value. Ammonium sulphate and urea concentrations effective in killing S. mansoni eggs and miracidia were about one to two orders of magnitude greater than the actual field application rates.     

Interpretation of microbial bioassays using kinetic parameters

A method analogous to that used for enzyme kinetics was applied to estimate maximum growth rates and oxygen depletion rates as measures of microbial toxicity. Selected toxicants including phenol, formalin and CuSO4 were tested against an axenic culture of E. coli and a mixed culture derived from soil to determine reproducibility of the test procedures. Optical density was used as a measure of growth over time. Oxygen uptake over a short term (less than 20 min.) and over extended time (greater than 4 hrs.) was also monitored. Microbial growth rate constants (g) and oxygen depletion rate constants (k) were calculated and the EC50 values for toxicity were taken as one half the maximum growth rate or oxygen depletion rates, respectively. The method proved a reasonable and reliable measure of toxicity.     

The effects of ammonium sulphate and urea upon egg hatching and miracidial survival of Schistosoma mansoni

Abstract

The effects of various concentrations of ammonium sulphate and urea on egg hatching and miracidial survival of S. mansoni were tested in order to determine if the use of these fertilizers in the ricelands of the Republic of Cameroon could affect the transmission of schistosomiasis. Results indicate that hatching of eggs and survival of miracidia varied with concentrations of tested chemicals and times of exposure. Exposure of S. mansoni eggs to 0.20%‐1.00% ammonium sulphate or to 0.50%‐4.00% urea reduced their ability to hatch and produce miracidia. A chemical concentration of 1.00% ammonium sulphate or 4.00% urea was found to be sufficient to produce complete inhibition of hatching. High concentrations of both chemicals not only inhibited miracidial emmergence but also may be ovocidal. Results obtained from miracidial survival tests indicated that LC₅, LC₅₀ and LC₉₅ values for ammonium sulphate were 0.07%, 0.80% and 10.61% after 2 hours of exposure; 0.03%, 0.16% and 0.90% after 4 hours of exposure; and 0.30%, 0.20% and 0.40% after 6 hours of exposure respectively. Similar statistical analyses revealed that the LC₅, LC₅₀ and LC₉₅ values for urea were 0.22%, 1.90% and 16.25% after 2 hours of exposure; 0.28%, 0.57% and 1.14% after 4 hours of exposure; and 0.13%, 0.27% and 0.57%” after 6 hours of exposure respectively. Although the two fertilizers exerted some adverse effects on S. mansoni eggs and miracidia at relatively high concentrations, neither of them was found to be of practical value. Ammonium sulphate and urea concentrations effective in killing S. mansoni eggs and miracidia were about one to two orders of magnitude greater than the actual field application rates.     

The effects of bayluscide and malathion on the survival of Schistosoma mansoni miracidia

Laboratory experiments were conducted to assess the toxic effects of bayluscide and malathion against Schistosoma mansoni miracidia. The results indicate that survival of miracidia varied with times of exposure and concentrations of tested chemicals. Statistical analyses reveal that LC5, LC50 and LC95 for bayluscide were 0.04 ppm, 0.06 ppm and 0.12 ppm after 2 hours of exposure; 0.02 ppm, 0.03 ppm and 0.06 ppm after 4 hours of exposure; and 0.01 ppm, 0.02 ppm and 0.04 ppm after 6 hours of exposure respectively. These data indicate that bayluscide is much more toxic to the first stage larvae of schistosomes than to snail intermediate hosts cited in the literature. Application of lower concentrations of molluscicide in the transmission sites is thus expected to curtail the survival of miracidia; therefore controlling schistosomiasis at relatively low costs. Such applications also reduce the risk of toxicity to non target organisms present in the aquatic environment. Statistical analysis of the results of tests using malathion gave LC5, LC50 and LC95 values of 83.38 ppm, 153.11 ppm and 245.85 ppm after 2 hours of exposure; and 76.86 ppm, 116.48 ppm and 172.04 ppm after 4 hours of exposure respectively. These data indicate that the use of malathion as an insecticide in tropical ecosystems may also affect the survival and viability of schistosome miracidia. Such uses could help reducing the risk of schistosomiasis transmission in these particular locations.