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Saprolegnia: Killer Cotton Dixie Daniels Department of Forestry, Wildlife, and Fisheries 22 April 2014

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Saprolegnia: Killer Cotton Dixie Daniels Department of Forestry, Wildlife, and Fisheries 22 April 2014 4/22/14 Saprolegnia: Killer Cotton Dixie Daniels Department of Forestry, Wildlife, and Fisheries 22 April 2014 S Overview S What is Saprolegnia? S How does it affect amphibians? S What makes Saprolegnia the most important factor in amphibian declines? Saprolegnia (van den Berg et al. 2013, Fernández-Benéitez 2008) S Saprolegnia spp. are common microorganisms in the class Oomycetes. S Image: Tom Volk, University of Wisconsin 1 4/22/14 Saprolegnia (Bruno et al. 1996, Willoughby 1994, Beakes 1982) S Diploid life cycle (2n) S Spores release asexual zoospores. S Once the zoophore finds suitable substrate, the sexual phase begins. S If the zoospore cannot find suitable substrate, it will continue producing more zoospores. Image: Bold et al. 1987 Mortality: Eggs (Fry et al. 2010, Petrisko et al. 2008) S Saprolegnia is the only genus of oomycete pathogens includes only water-borne organisms S Infection = mortality S Parasitizes eggs Image: American Phytopathological Society Mortality: Larvae (Romansic 2009) S Comparative study of the effects of S. ferax on S Pseudacris regilla (Pacific treefrog) S Rana cascadae (Cascades frog) S Ambystoma macrodactylum (long-toed salamander) S Rana aurora (red-legged frog) 2 4/22/14 Mortality: Larvae (Romansic 2009) S A one week exposure to S. ferax 3 4/22/14 Amphibian Declines (Kiesecker et al. 1995, Fry et al. 2010) S Usually considered a secondary pathogen, in concert with other issues such as chytrid fungus or UV-B radiation exposure, Saprolegnia infection is the fatal blow. S Under the right circumstances, Saprolegnia can act as a primary pathogen, causing mycosis and mortality. Prolonged11050 Effects?Ecology: Kiesecker and Blaustein Proc. Natl. Acad. Sci. USA 92 (1995) (Blaustein et al. 1994) Experiment 2 was conducted at Three Creeks Lake simul- directly into the lake. Thus, in this experiment, embryos were taneously with the tests conducted for experiment 1 at Three exposed t,o all three sunlight regimes and natural levels of Creeks Lake. In experiment 2, we used procedures identical to Saprolegnia. those in experiment 1 except that enclosures were placed The experiments ended when all embryos either hatched or died. Survival was measured as the proportion of hatchlings 100 produced per enclosure. The proportion of hatchlings pro- duced per enclosure (survivorship through hatching) was S assessed using analysis of variance (ANOVA) to test for Mean egg survival rates 80 differences among the treatments. are dropping drastically. RESULTS 60 In experiment 1, the ANOVA indicated a significant UV-B S Fewer eggs mean fewer effect by itself. However, the effect is secondary to the 40 Table 1. Experiment 1: ANOVA of hatching success in three adults! amphibian species 20 Source of variation MS df F P B. boreas Lost Lake 0No fungus Fungus No fungus Fungus UV 0.034 2 55.920 <0.001 Fungus 0.574 1 938.024 <0.001 Lost Lake Three Creeks UV x fungus 0.043 2 69.484 <0.001 Bufo Error 0.001 18 100 Three Creeks Image: Kiesecker et al. UV 0.023 2 16.958 <0.001 Fungus 0.493 1 363.966 <0.001 eo80 UV x fungus 0.025 2 18.455 <0.001 Error 0.001 18 R. cascadae Small Lake 60 UV 0.014 2 7.544 0.004 Fungus 0.658 1 360.660 <0.001 UV x fungus 0.018 2 10.079 <0.001 40 Error 0.002 18 Three Creeks 00 UV 0.032 2 15.879 <0.001 Fungus 0.748 1 368.494 <0.001 UV x fungus 0.027 2 13.529 <0.001 Error 0.002 18 H. regilla No fungus Fungus No fungus Fungus Small Lake SummarySmall Lake Three Creeks UV 0.001 2 0.977 0.396 Rana Fungus 0.137 1 266.734 <0.001 UV x fungus 0.001 2 2.185 0.141 100 Error 0.001 18 Three Creeks UV <0.001 2 0.300 0.746 80 Fungus 0.060 1 150.834 <0.001 UV x fungus <0.001 2 0.018 0.982 Error <0.001 18 S Saprolegnia is a parasitic microorganism60 A preliminary analysis indicated no significant block effects (i.e., no differences between temperature or other variables among blocks). Therefore, the block and error terms were pooled for remaining tests 40 (18). Post hoc comparisons (Tukey Test) (18) were performed to test S It kills amphibian eggs, larvae, and adults for differences between means among the six regimes. Temperatures were taken within enclosures for each species in each treatment. Mean temperatures (and ANOVAs) are given for each species at each site S 20 for the unfiltered, UV-B-transmitting, and UV-B-blocking regimes When other factors weaken amphibian with and without the fungus respectively: Bufo at Lost Lake = 8.3° C, 8.7° C, 8.8° C, 9.1° C, 8.6° C, and 8.5° C, F5,18 = 0.822, MS = 0.034, P = populations, Saprolegnia infection can be a 0.649; Bufo at Three Creeks = 12.0° C, 12.9° C, 13.2° C, 12.1° C, 12.8° C, 0No fungus Fungus No fungus Fungus and 12.6° C, F5,18 = 1.294, MS = 0.039, P = 0.160; Rana at Small Lake devastating blow to species = 10.8° C, 11.3° C, 10.7° C, 11.3° C, 10.7° C, and 10.30C, F5,18 = 1.061, MS Small Lake Three Creeks = 0.034, P = 0.464; Rana at Three Creeks = 10.90C, 12.60C, 11.5° C, Hyla 11.4° C, 11.6° C, and 12.5° C, F5,18 = 2.237, MS = 0.043, P = 0.227; Hyla at Small Lake = 11.2° C, 10.90C, 11.6° C, 11.4° C, 10.7° C, and 11.9° C, FIG. 1. Experiment 1: Effects of UV-B radiation and manipulated F5,18 - 1.561, MS = 0.005, P = 0.271; Hyla at Three Creeks = 12.3° C, amounts of S. ferax on hatching success (x ± SE) in B. boreas, R. 13.8° C, 14.3° C, 13.6° C, 13.7° C, and 12.9° C, F5,18 = 0.798, MS = 0.006, cascadae, and H. regilla. *, UV-B-blocking filter; n, UV-B- P = 0.667. MS, mean square; F, F statistic [with degrees of freedom transmitting filter; 0, no filter. (df)]; P, probability. 4 4/22/14 Literature Cited S van den Berg, A. H., D. Mclaggan, J. Dieguez-Uribeond, and P. van West. 2013. The impact of the water moulds Saprolegnia diclina and Saprolegnia parasitica on natural ecosystems and the aquaculture industry. Fungal Biology Reviews 27:33–42. S Fernandez-Beneitez, M. J., M. E. Ortiz-Santaliestra, M. Lizana, and J. Dieguez-Uribeondo. 2008. Saprolegnia diclina: another species responsible for the emergent disease “Saprolegnia infections” in amphibians. Fems Microbiology Letters 279:23–29. S Bruno, D. W., and T. T. Poppe. 1996. A colour atlas of salmonid diseases. Academic press. S Willoughby, L. G. 1994. Fungi and fish diseases. Pisces Press. S Beakes, G. 1983. A comparative account of cyst coat ontogeny in saprophytic and fish-lesion (pathogenic) isolates of the Saprolegnia diclina-parasitica complex. Canadian Journal of Botany 61:603–625. S Fry, W. E., and N. J. Grünwald. 2010. Introduction to Oomycetes. Plant Health Instructor. S Petrisko, J. E., C. A. Pearl, D. S. Pilliod, P. P. Sheridan, C. F. Williams, C. R. Peterson, and R. B. Bury. 2008. Saprolegniaceae identified on amphibian eggs throughout the Pacific Northwest, USA, by internal transcribed spacer sequences and phylogenetic analysis. Mycologia 100:171–180. S Romansic, J. M., K. A. Diez, E. M. Higashi, J. E. Johnson, and A. R. Blaustein. 2009. Effects of the pathogenic water mold Saprolegnia ferax on survival of amphibian larvae. Diseases of Aquatic Organisms 83:187–193. S Kiesecker, J. M., and A. R. Blaustein. 1995. Synergism between UV-B radiation and a pathogen magnifies amphibian embryo mortality in nature. Proceedings of the National Academy of Sciences 92:11049–11052. S Blaustein, A. R., D. Grant Hokit, R. K. O’Hara, and R. A. Holt. 1994. Pathogenic fungus contributes to amphibian losses in the pacific northwest. Biological Conservation 67:251–254. Questions? 5 .
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