1. The Effects of Triclosan on Five Marine Algae Species Melanie N. McHenry Department of Biology, Marine Science Program, Jackson State University, P.O. Box 18540, 1400 J.R. Lynch Street, Jackson, Mississippi, USA

8. Background Information Cont. Triclosan Levels in Waste Water Waste water Levels ( µ g/L) Source Primary Effluent 3.4 - 8.0 McAvoy, DC et al. 2002 Final Effluent: Activated Sludge Treatment 0.20 - 0.41 Weigel S. et al. : 2004 Trickling Filter Treatment 1.6 - 2.7 Switzerland Effluents 42 - 213 mg/L Balmer 2005

10. Algae Used in this Study Tetraselmis chuii Nannochloropsis oculata Chaetocerus gracilis Isochrysis galbana Rhodomonas salina

11. Algae Information Algae Class Size (µm) Motile Flagella Isochrysis Prymnesiophyceae 4 – 8 yes 2 flagella Tetraselmis Chlorophyceae 12 - 14 4 flagella Nannochloropsis Eustigmatophyceae 1.5 - 2.5 no none Rhodomonas Cryptophyceae 16 – 30 yes Chaetocerus Baccillariophyceae 12 - 14 no

12. The Effects of Triclosan on Various Organisms (Balmer 2005, Russel 2004, Orvos et al. 2002) Organism L(E) 50 Algae Freshwater algae Anabaena flos-aquae 0.97 µg/L Scenedesmus subspicatus 2. 8 µg/L Selenastrum capricornutum 4.46 µg/L Navicula pelliculosa 19.1 µg/L Marine Algae Skeletonema costatum > 66.0 µg/L Fish Danio rerio and Oryzais latipes < 300 µg/L Pimephales promelas 260 µg/L Lepomis macrochirus 370 µg/L Seamonkeys Artemia 2000 µg/L Bacteria Methicillin-resistant Staphylococci 0.1 – 2 mg/L E. coli and K. pueumoniae 5,000 µg/L

14. Experimental Design Treatment (ug/L) Replicate # Temperature (average) Salinity Light (lux) Control 0 3 25 o C 25ppt 2,200 1 0.2 3 25 o C 25ppt 2,200 2 1.0 3 25 o C 25ppt 2,200 3 5.0 3 25 o C 25ppt 2,200 4 25.0 3 25 o C 25ppt 2,200 5 125 3 25 o C 25ppt 2,200 6 250 3 25 o C 25ppt 2,200 7 500 3 25 o C 25ppt 2,200 Solvent Control 7.2% Ethyl Alcohol 3 25 o C 25ppt 2,200

17. Isochrysis galbana

18. Tetraselmis chuii

19. Nannochloropsis oculata

20. Rhodomonas salina

21. Chaetocerus gracilis

23. Isochrysis galbana

24. Isochrysis growth rate ANOVA table 24 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 36.61077037 3.66107704 369.77 < 0.0001 Error 16 0.15841481 0.00990093 Total 26 36.76918519 R-Square Coefficient Var Root MSE YIELD Mean 0.995692 13.61679 0.099503 0.730741

25. Means with the same letter are not significantly different t Grouping Mean N Treatment A 1.85667 3 Control B 1.65000 3 Treatment 1 C 1.29333 3 Treatment 2 D 1.02333 3 Treatment 3 E 0.61333 3 Treatment 4 F 0.31000 3 Treatment 5 G -0.01000 3 Treatment 6 H -203333 3 Treatment 7 A 1.87333 3 Treatment 8

26. Isochrysis growth rate ANOVA table 48 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 8.75095556 0.87509556 198.57 < 0.0001 Error 16 0.07051111 0.00440694 Total 26 8.82146667 R-Square Coefficient Var Root MSE YIELD Mean 0.992007 12.11893 0.066385 0.547778

27. Means with the same letter are not significantly different t Grouping Mean N Treatment A 1.16667 3 Control B 0.91000 3 Treatment 1 C 0.72000 3 Treatment 2 D 0.57667 3 Treatment 3 E D 0.49000 3 Treatment 4 E 0.42333 3 Treatment 5 F 0.23667 3 Treatment 6 G -0.8000 3 Treatment 7 A 1.20667 3 Treatment 8

28. Isochrysis growth rate ANOVA table 72 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 2.49982222 0.24998222 1097.48 <0.0001 Error 16 0.00364444 0.00022778 Total 26 2.50346667 R-Square Coefficient Var Root MSE YIELD Mean 0.998544 2.538893 0.015092 0.594444

29. Means with the same letter are not significantly different t Grouping Mean N Treatment A 0.9600 3 Control B 0.8500 3 Treatment 1 C 0.82333 3 Treatment 2 D 0.75667 3 Treatment 3 E 0.67333 3 Treatment 4 F 0.56667 3 Treatment 5 G 0.48667 3 Treatment 6 H 0.31667 3 Treatment 7 I -0.08333 3 Treatment 8

30. Isochrysis growth rate ANOVA table 96 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 1.23659259 0.12365926 763.15 < <0.0001 Error 16 0.00259259 0.00016204 Total 26 1.23918519 R-Square Coefficient Var Root MSE YIELD Mean 0.997908 2.197527 0.012729 0.579259

31. Means with the same letter are not significantly different t Grouping Mean N Treatment A 0.83333 3 Control A 0.82667 3 Treatment 1 B 0.73333 3 Treatment 2 C 0.67333 3 Treatment 3 D 0.59333 3 Treatment 4 E 0.52667 3 Treatment 5 F 0.48333 3 Treatment 6 G 0.43333 3 Treatment 7 H 0.1100 3 Treatment 8

32. Tetraselmis chuii

33. Tetraselmis growth rate ANOVA table 24 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 10.17803704 1.01780370 81.79 < 0.0001 Error 16 0.1991037 0.01244398 Total 26 10.37714074 R-Square Coefficient Var Root MSE YIELD Mean 0.980813 55.36618 0.111553 0.201481

34. Means with the same letter are not significantly different t Grouping Mean N Treatment A 0.68000 3 Control B A 0.60667 3 Treatment 1 B 0.51000 3 Treatment 2 B 0.46333 3 Treatment 3 C 0.24333 3 Treatment 4 C 0.14667 3 Treatment 5 D -0.15667 3 Treatment 6 E -1.36667 3 Treatment 7 A 0.68667 3 Treatment 8

35. Tetraselmis growth rate ANOVA table 48 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 1.02720000 0.10272000 2.89 0.0287 Error 16 0.56946667 0.03559167 Total 26 1.59666667 R-Square Coefficient Var Root MSE YIELD Mean 0.643340 44.68205 0.188658 0.422222

36. Means with the same letter are not significantly different t Grouping Mean N Treatment A 0.6200 3 Control A 0.5933 3 Treatment 1 B A 0.5567 3 Treatment 2 B A 0.5400 3 Treatment 3 B A 0.4567 3 Treatment 4 B 0.4667 3 Treatment 5 B C 0.2367 3 Treatment 6 C -0.0267 3 Treatment 7 A 0.3467 3 Treatment 8

37. Tetraselmis growth rate ANOVA table 72 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 1.05683704 0.10568370 3.38 0.0149 Error 16 0.50061481 0.03128843 Total 26 1.55745185 R-Square Coefficient Var Root MSE YIELD Mean 0.678568 30.71321 0.176885 0.575926

38. Means with the same letter are not significantly different t Grouping Mean N Treatment A 0.8567 3 Control B A 0.7700 3 Treatment 1 B A 0.6933 3 Treatment 2 B A C 0.6300 3 Treatment 3 B A C 0.5733 3 Treatment 4 B C 0.5100 3 Treatment 5 C 0.3767 3 Treatment 6 0.1600 3 Treatment 7 A 0.6133 3 Treatment 8

39. Tetraselmis growth rate ANOVA table 96 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 0.77628889 0.07762889 3.08 0.0221 Error 16 0.40377778 0.02523611 Total 26 1.18006667 R-Square Coefficient Var Root MSE YIELD Mean 0.657835 21.69543 0.158859 0.73222

40. Means with the same letter are not significantly different t Grouping Mean N Treatment A 0.8967 3 Control A 0.8700 3 Treatment 1 A 0.8500 3 Treatment 2 A 0.8067 3 Treatment 3 A 0.7867 3 Treatment 4 A 0.7433 3 Treatment 5 A 0.6600 3 Treatment 6 A 0.3067 3 Treatment 7 B 0.6700 3 Treatment 8

41. Nannochloropsis oculata

42. Nannochloropsis growth rate ANOVA table 24 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 5.89219259 0.58921926 45.75 < 0.0001 Error 16 0.20605926 0.0128787 Total 26 6.09825185 R-Square Coefficient Var Root MSE YIELD Mean 0.966210 17.30140 0.113484 0.655926

43. Means with the same letter are not significantly different t Grouping Mean N Treatment A 1.32667 3 Control B 0.92667 3 Treatment 1 C B 0.85000 3 Treatment 2 C 0.71000 3 Treatment 3 D 0.51333 3 Treatment 4 E 0.29333 3 Treatment 5 F -0.08333 3 Treatment 6 F 0.09333 3 Treatment 7 A 1.27333 3 Treatment 8

44. Nannochloropsis growth rate ANOVA table 48 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 4.89813333 0.48981333 4.00 0.0069 Error 16 1.95913333 0.12244583 Total 26 6.85726667 R-Square Coefficient Var Root MSE YIELD Mean 0.714298 32.77111 0.349923 1.067778

45. Means with the same letter are not significantly different t Grouping Mean N Treatment A 1.6300 3 Control B A 1.5067 3 Treatment 1 B A 1.3900 3 Treatment 2 B A C 1.1600 3 Treatment 3 B A C 1.1033 3 Treatment 4 B C 0.9500 3 Treatment 5 C 0.5600 3 Treatment 6 0.2100 3 Treatment 7 A 1.1000 3 Treatment 8

46. Nannochloropsis growth rate ANOVA table 76 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 4.89813333 0.48981333 4.00 0.0069 Error 16 1.95913333 0.12244583 Total 26 6.85726667 R-Square Coefficient Var Root MSE YIELD Mean 0.714298 32.77111 0.349923 1.067778

47. Means with the same letter are not significantly different t Grouping Mean N Treatment 1.63000 3 Control B 1.5067 3 Treatment 1 B 1.3900 3 Treatment 2 B C 1.1600 3 Treatment 3 B C 1.1033 3 Treatment 4 B C 1.1000 3 Treatment 5 D C 0.9500 3 Treatment 6 D 0.5600 3 Treatment 7 B A C 0.2100 3 Treatment 8

48. Nannochloropsis growth rate ANOVA table 96 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 2.04006667 0.20400667 20.12 <0.0001 Error 16 0.16220000 0.01013750 Total 26 2.20226667 R-Square Coefficient Var Root MSE YIELD Mean 0.926349 19.52945 0.100685 0.515556

49. Means with the same letter are not significantly different t Grouping Mean N Treatment A 0.73667 3 Control B A 0.7100 3 Treatment 1 B A 0.66333 3 Treatment 2 B A 0.63000 3 Treatment 3 B C 0.54667 3 Treatment 4 C 0.41000 3 Treatment 5 D 0.23333 3 Treatment 6 E -0.06333 3 Treatment 7 A 0.77333 3 Treatment 8

50. Rhodomonas salina

51. Rhodomonas growth rate ANOVA table 24 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 3.28800000 0.32880000 31.99 <0.0001 Error 16 0.16446667 0.01027917 Total 26 3.45246667 R-Square Coefficient Var Root MSE YIELD Mean 0.952363 12.48257 0.101386 0.812222

52. Means with the same letter are not significantly different t Grouping Mean N Treatment A 1.13667 3 Control B A 1.09333 3 Treatment 1 B C 0.95333 3 Treatment 2 C 0.91667 3 Treatment 3 D C 0.78667 3 Treatment 4 D 0.69333 3 Treatment 5 E 0.34667 3 Treatment 6 F 0.14333 3 Treatment 7 A 1.24000 3 Treatment 8

53. Rhodomonas growth rate ANOVA table 48 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 1.68814815 0.16881481 2.29 0.0675 Error 16 1.17988148 0.07374259 Total 26 2.86802963 R-Square Coefficient Var Root MSE YIELD Mean 0.588609 35.55775 0.271556 0.763704

54. Means with the same letter are not significantly different t Grouping Mean N Treatment A 1.1267 3 Control A 1.05333 3 Treatment 1 A 0.7267 3 Treatment 2 B A 0.8233 3 Treatment 3 B A 0.7600 3 Treatment 4 B A 0.7267 3 Treatment 5 B C 0.5567 3 Treatment 6 C 0.2333 3 Treatment 7 B A 0.8667 3 Treatment 8

55. Rhodomonas growth rate ANOVA table 72 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 1.36481481 0.13648148 3.80 0.0087 Error 16 0.57394815 0.03587175 Total 26 1.93876296 R-Square Coefficient Var Root MSE YIELD Mean 0.703962 27.97460 0.189398 0.677037

56. Means with the same letter are not significantly different t Grouping Mean N Treatment A 0.9300 3 Control A 0.8933 3 Treatment 1 A 0.8367 3 Treatment 2 A 0.8100 3 Treatment 3 B A 0.7167 3 Treatment 4 B A 0.6300 3 Treatment 5 B C 0.4567 3 Treatment 6 C 0.1800 3 Treatment 7 B A 0.6400 3 Treatment 8

57. Rhodomonas growth rate ANOVA table 96 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 0.97977037 0.09797704 3.17 0.0196 Error 16 0.49503704 0.03093981 Total 26 1.47480741 R-Square Coefficient Var Root MSE YIELD Mean 0.664338 29.90695 0.175897 0.599148

58. Means with the same letter are not significantly different t Grouping Mean N Treatment A 0.8600 3 Control B A 0.7567 3 Treatment 1 B A C 0.6900 3 Treatment 2 B A C 0.6567 3 Treatment 3 B A C 0.6000 3 Treatment 4 B C 0.5467 3 Treatment 5 D C 0.4167 3 Treatment 6 D 0.1667 3 Treatment 7 B A C 0.6000 3 Treatment 8

59. Chaetocerus gracilis

60. Chaetocerus growth rate ANOVA table 24 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 35.12774815 3.51277481 34.51 < 0.0001 Error 16 1.62859259 0.010178704 Total 26 36.75634074 R-Square Coefficient Var Root MSE YIELD Mean 0.955692 32.94112 0.319041 0.968519

61. Means with the same letter are not significantly different t Grouping Mean N Treatment A 2.0967 3 Control B A 1.7767 3 Treatment 1 B A 1.5867 3 Treatment 2 B C 1.2633 3 Treatment 3 D C 0.9100 3 Treatment 4 D 0.6700 3 Treatment 5 E 0.0433 3 Treatment 6 F -1.6967 3 Treatment 7 A 2.0667 3 Treatment 8

62. Chaetocerus growth rate ANOVA table 48 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 4.85472593 0.48547259 2.86 0.0298 Error 16 2.71705926 0.16981620 Total 26 7.57178519 R-Square Coefficient Var Root MSE YIELD Mean 0.641160 50.82853 0.412088 0.810741

63. Means with the same letter are not significantly different t Grouping Mean N Treatment A 1.3067 3 Control B A 1.1767 3 Treatment 1 B A 1.0633 3 Treatment 2 B A 0.9633 3 Treatment 3 B A 0.9000 3 Treatment 4 B A 0.8033 3 Treatment 5 B 0.5367 3 Treatment 6 C -0.2133 3 Treatment 7 B A 0.7600 3 Treatment 8

64. Chaetocerus growth rate ANOVA table 72 hours Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 1.06761482 0.10676148 3.04 0.0233 Error 16 0.56248148 0.03515509 Total 26 1.63009630 R-Square Coefficient Var Root MSE YIELD Mean 0.654940 29.31336 0.187497 0.639630

65. Means with the same letter are not significantly different t Grouping Mean N Treatment A 0.9167 3 Control B A 0.8300 3 Treatment 1 B A 0.7700 3 Treatment 2 B A 0.6967 3 Treatment 3 B A 0.6500 3 Treatment 4 B A 0.6200 3 Treatment 5 B 0.5600 3 Treatment 6 C 0.5233 3 Treatment 7 B 0.5600 3 Treatment 8

66. Chaetocerus growth rate ANOVA table 96 hour Source of Variation Degrees of Freedom Sum of Squares Mean Square F-Value Pr > F Model 10 0.65040000 0.06504000 2.63 0.0412 Error 16 0.39600000 0.02475000 Total 26 1.04640000 R-Square Coefficient Var Root MSE YIELD Mean 0.621560 27.76259 0.157321 0.56667

67. Means with the same letter are not significantly different t Grouping Mean N Treatment A 0.7800 3 Control B A 0.7433 3 Treatment 1 B A C 0.6733 3 Treatment 2 B A C 0.5967 3 Treatment 3 B A C 0.5533 3 Treatment 4 B D C 0.5033 3 Treatment 5 D C 0.4500 3 Treatment 6 D 0.2367 3 Treatment 7 B A C 0.5633 3 Treatment 8

69. Percent Inhibition: Isochrysis Percent Inhibition Initial Day 1 Day 2 Day 3 Day 4 Control (0µg/L) n/a n/a n/a n/a n/a Treatment 1 (0.2µg/L) n/a 11.27 22.10 11.62 12.28 Treatment 2 (1.0µg/L) n/a 30.40 38.49 21.35 19.11 Treatment 3 (5.0µg/L) n/a 44.80 50.69 29.52 28.48 Treatment 4 (25µg/L) n/a 66.73 58.22 40.98 36.59 Treatment 5 (125µg/L) n/a 83.19 63.75 49.47 41.86 Treatment 6 (250µg/L) n/a 100.60 79.30 67.29 47.90 Treatment 7 (500µg/L) n/a 208.44 166.54 108.72 86.96 Treatment 8 (ethyl alcohol control) n/a -0.65 -3.10 14.09 0.72

70. Percent Inhibition: Tetraselmis Percent Inhibition Initial Day 1 Day 2 Day 3 Day 4 Control (0µg/L) n/a n/a n/a n/a n/a Treatment 1 (0.2µg/L) n/a 12.55 4.13 14.98 4.20 Treatment 2 (1.0µg/L) n/a 26.30 7.89 20.47 7.27 Treatment 3 (5.0µg/L) n/a 33.20 14.67 30.24 10.73 Treatment 4 (25µg/L) n/a 64.64 25.66 35.64 14.12 Treatment 5 (125µg/L) n/a 79.14 26.39 42.91 18.79 Treatment 6 (250µg/L) n/a 122.51 71.78 61.49 33.29 Treatment 7 (500µg/L) n/a 432.19 288.66 90.46 78.05 Treatment 8 (ethyl alcohol control) n/a -1.59 -0.54 -2.60 0.57

71. Percent Inhibition: Nannochloropsis Percent Inhibition Initial Day 1 Day 2 Day 3 Day 4 Control (0µg/L) n/a n/a n/a n/a n/a Treatment 1 (0.2µg/L) n/a 30.70 10.46 4.95 2.54 Treatment 2 (1.0µg/L) n/a 35.82 16.51 7.85 4.46 Treatment 3 (5.0µg/L) n/a 46.84 30.71 12.78 6.76 Treatment 4 (25µg/L) n/a 60.74 34.51 20.63 13.38 Treatment 5 (125µg/L) n/a 77.83 44.57 43.61 20.90 Treatment 6 (250µg/L) n/a 106.07 75.01 55.89 39.63 Treatment 7 (500µg/L) n/a 89.70 93.71 95.22 95.37 Treatment 8 (ethyl alcohol control) n/a -49.15 -21.24 -9.21 -9.08

72. Percent Inhibition: Rhodomonas Percent Inhibition Initial Day 1 Day 2 Day 3 Day 4 Control (0µg/L) n/a n/a n/a n/a n/a Treatment 1 (0.2µg/L) n/a 4.10 22.20 4.82 14.02 Treatment 2 (1.0µg/L) n/a 16.60 28.75 10.38 21.23 Treatment 3 (5.0µg/L) n/a 20.02 30.98 14.93 22.55 Treatment 4 (25µg/L) n/a 31.18 34.81 25.45 33.28 Treatment 5 (125µg/L) n/a 39.50 36.92 32.12 36.87 Treatment 6 (250µg/L) n/a 68.58 57.83 60.78 56.97 Treatment 7 (500µg/L) n/a 87.38 92.06 90.54 91.51 Treatment 8 (ethyl alcohol control) n/a -13.41 -9.31 1.15 -1.61

73. Percent Inhibition: Chaetocerus Percent Inhibition Initial Day 1 Day 2 Day 3 Day 4 Control (0µg/L) n/a n/a n/a n/a n/a Treatment 1 (0.2µg/L) n/a 0.73 11.37 10.10 9.02 Treatment 2 (1.0µg/L) n/a 11.43 21.11 16.12 18.73 Treatment 3 (5.0µg/L) n/a 29.20 27.17 25.50 26.78 Treatment 4 (25µg/L) n/a 49.11 31.30 28.84 32.09 Treatment 5 (125µg/L) n/a 62.57 41.25 31.98 37.08 Treatment 6 (250µg/L) n/a 96.98 65.26 46.86 46.52 Treatment 7 (500µg/L) n/a 181.22 125.25 95.00 83.40 Treatment 8 (ethyl alcohol control) n/a -15.51 0.30 8.33 -1.95

75. Isochrysis galbana

76. The time Period and its corresponding EC 50 value Time Period EC 50 value Day 1 2.10 μg/mL Day 2 2.44 μg/mL Day 3 3.97 μg/mL Day 4 4.81 μg/mL

77. Tetraselmis chuii

78. The time period and its corresponding EC 50 value Time Period EC 50 value Day 1 10.0 μg/mL Day 2 77.7 μg/mL Day 3 83.8 μg/mL Day 4 >500 μg/mL

79. Nannochloropsis oculata

80. The time period and its corresponding EC 50 value Time Period EC 50 value Day 1 2.10 μg/mL Day 2 2.44 μg/mL Day 3 3.97 μg/mL Day 4 4.81 μg/mL

81. Rhodomonas salina

82. The time period and its corresponding EC 50 value Time Period EC 50 value Control (0µg/L) 109 μg/mL Treatment 1 (0.2µg/L) 98 μg/mL Treatment 2 (1.0µg/L) 168 μg/mL Treatment 3 (5.0µg/L) 128 μg/mL

83. Chaetocerus gracilis

84. The time period and its corresponding EC 50 value Time Period EC 50 value Control (0µg/L) 46.2 μg/mL Treatment 1 (0.2µg/L) 68 μg/mL Treatment 2 (1.0µg/L) 183 μg/mL Treatment 3 (5.0µg/L) 162 μg/mL

85. Discussion: Compare Marine to Freshwater Algae Time Period Marine Algae Species EC 50 values Isochrysis Nannochloropsis Rhodomonas Chaetocerus Tetraselmis 24hr 10.8 6.3 109 46.2 10.0 48hr 68.5 72 98 68 77.7 72hr 182 165 168 183 83.8 96hr 164 360 128 162 >500 Freshwater Algae Species L(E) 50 values Navicula pelliculosa Selenastrum capricornutum Scenedesmus subspicatus Skeletonema costatum Anabaena flosaquae 96hr 19.1 µg/L 4.46 µg/L 2. 8 µg/L > 66.0 µg/L 0.97 µg/L

87. Discussion: Comparison of EC 50 and L(E) 50 Values of Other Organisms Time Period Marine Algae Species EC 50 values Isochryis galbana Nannochloropsis oculata Rhodomonas salina Chaetocerus gracilis Tetraselmis chuii 24hr 10.8 6.3 109 46.2 10.0 48hr 68.5 72 98 68 77.7 72hr 182 165 168 183 83.8 96hr 164 360 128 162 >500 Organisms L(E) 50 values Fish Seamonkeys Bacteria Danio rerio Oryzais latipes Artemia Staphylococci Pimephales promelas Lepomis macrochirus E. coli K. pueumoniae Pseudomonas aeruginosa < 300 µg/L 2000 µg/L 100-300 µg/L 260 µg/L 370 µg/L 5,000 µg/L 300,000-1,000,000 µg/L

90. Discussion: Comparison of the EC 50 Values among the Algae Species Time Period Marine Algae Species EC 50 values Isochryis galbana Nannochloropsis oculata Rhodomonas salina Chaetocerus gracilis Tetraselmis chuii 24hr 10.8 6.3 109 46.2 10.0 48hr 68.5 72 98 68 77.7 72hr 182 165 168 183 83.8 96hr 164 360 128 162 >500

91. Discusssion: Visual comparison of the EC 50 (µg/L) values

94. Acknowledgements This research was supported, in part, by a grant from the National Oceanic and Atmospheric Administration grant # NA17AE1626, Subcontract # 27-0629-017, through the Environmental Cooperative Science Center at Florida A&M University to Jackson State University. M. McHenry was supported through the National Institutes of Health funded Research Initiative for Scientific Enhancement (RISE) grant awarded to Jackson State University.