From Silent Spring to Silent Night: A Tale of Toads and Men Ubuntu ungamntu ngabanye abantu Ubuntu ungamntu ngabanye abantu Xhosa proverb "People are people through other people". LABORATORY FOR INTEGRATIVE STUDIES IN AMPHIBIAN BIOLOGY Funded by: The National Science Foundation, Novartis, Syngenta Crop Protection, Ecorisk, the World Wildlife Fund, the Alton Jones Foundation, the Homeland Foundation, the Rose Foundation, Park-Water Company, the Biofaculty Award [UCB], the Distinguished Mentor Award (UCB), the Distinguished Teaching Award (UCB), the Mitchell Kapor Foundation, the David Foundation, the Cornell-Douglas Foundation, the Wallace Global Fund, the Class of ‘43 Endowed Chair (UCB), the Toxic Substances Research and Teaching Program (UCB), Hewlett Packard, the Biology Fellows Program (Howard Hughes Medical Institute), the McNair Scholars Program (UCB), the Amgen Scholars Program (UCB), and the Mentored Research fellowship program (UCB), Marisla Foundation, Beyond Pesticides LABORATORY FOR INTEGRATIVE STUDIES IN AMPHIBIAN BIOLOGY Funded by: The National Science Foundation, Novartis, Syngenta Crop Protection, Ecorisk, the World Wildlife Fund, the Alton Jones Foundation, the Homeland Foundation, the Rose Foundation, Park-Water Company, the Biofaculty Award [UCB], the Distinguished Mentor Award (UCB), the Distinguished Teaching Award (UCB), the Mitchell Kapor Foundation, the David Foundation, the Cornell-Douglas Foundation, the Wallace Global Fund, the Class of ‘43 Endowed Chair (UCB), the Toxic Substances Research and Teaching Program (UCB), Hewlett Packard, the Biology Fellows Program (Howard Hughes Medical Institute), the McNair Scholars Program (UCB), the Amgen Scholars Program (UCB), and the Mentored Research fellowship program (UCB), Marisla Foundation, Beyond Pesticides LABORATORY FOR INTEGRATIVE STUDIES IN AMPHIBIAN BIOLOGY Aaron Vonk Ahmad Ali Stuart Huy Ngao Maggy Mendoza Atif Collins Victoria Ngo Kelly Haston Nigel Noriega Varinder Pharenga Roger Liu Sarah Chui Travis Brown Melissa Lee Anu Devi Sona gill Vineet Jindahl Emily Chan Marquez Patricia Hom Gwynne JohnstonAdrian Brunner-Brown Lynette Chen Karen Chan Isabel Hsu Sherrie Gallipeau Jeffrey Lee Mable Tsui Young Kim-Parker Nadir Yehya Diana Reyes Erin Conner Paola Case Catherine Haeffele Porntip Propunpoj Youssra Marjoua Duc Chung Kristin Trott Nini Mai Anhthu Hoang John Parker Vicky Khoury Farah Syed Miriam Olivera Anna Feldman Laura Meehan Anne Narayan LABORATORY FOR INTEGRATIVE STUDIES IN AMPHIBIAN BIOLOGY Martin Hansen Cara He Sherrie Gallipeau Fay Pon Susan Luong Maggie Tsang Ngoc Mai Nguyen KarenZhang Diana Salazar Nisha Balabhadra Betsy Rosales Julia Chow Janiel Ankin Chin Tai Flor Gowans Alejandra Cabrera LABORATORY FOR INTEGRATIVE STUDIES IN AMPHIBIAN BIOLOGY Cara He Karen Zhang Maxwell Nguyen Alina Nguyen Clara Pavesi Krieger Alejandra Rivas Adonis Rivie Maggie Tsang Andrea Meraz Julia Chow Nazher Indris Dax Vivid Bryanna Perez Jibril Kyser Adriana Lawson LABORATORY FOR INTEGRATIVE STUDIES IN AMPHIBIAN BIOLOGY Cara He Maxwell Nguyen Andrea Meraz Alina Nguyen Nazher Indris Adonis Rivie Bryanna Perez Chris Harrod Jhoselyn Pineda Seunmin Yoon Ashlie Barillas LABORATORY FOR INTEGRATIVE STUDIES IN AMPHIBIAN BIOLOGY Verenice Bravo Axenya Kachen Shreya Shankar Adriana Martinez Leena Bakshi Your life is your story. Live it well…Tell it well. (tyrone B hayes, on walking 3 hours to work) Male Female Estradiol Testosterone Control 1 2 3 4 5 6 Horm Dep Color Change E2 + T E2 only HYPOTHESIS 1: T SR E2 HYPOTHESIS 2: T AR E2 ER HYPOTHESIS 3: T E2 ER Hyperolius viridiflavus spp. CON T E2 DHT TESTOSTERONE CONTROL TESTOSTERONE + TAMOXIFEN OH OH Aromatase ER O HO Testosterone Estradiol 1.2 1.0 weight) 0.8 activity body 0.6 0.4 Aromatase 0.2 conversion/ 0.0 (% (% -AI +AI -AI +AI H. variabilus H. argus Y R N C P G D R E U O E R S C E O R N L I I N U E S OH HO OH HO 2OHE2 HO OH 4OHE2 O O OH ER O HO HO ADSD E1 E2 OSO 4 OH HO O SO E2-17SO4 4 E2-3SO4 OH HO OH HO 2OHE2 HO OH 4OHE2 O O OH ER O HO HO ADSD E1 E2 OSO 4 OH HO O SO E2-17SO4 4 E2-3SO4 OH HO OH HO 2OHE2 HO OH 4OHE2 O O OH ER O HO HO ADSD E1 E2 OSO 4 OH HO O SO E2-17SO4 4 E2-3SO4 Control Estradiol Ethynyl-E2 DES DDT Estrogen + Tamoxifen ATRAZINE INH THE U.HS. ATRAZINE INHIBITED GROWTH OF THE VOICE BOX IN MALES Testes Ovaries Testis Ovaries OH O Testosterone T OH OH Aromatase O HO Testosterone Estrogen ATRAZINE E 6 5 4 (ng/mL) 3 2 testosterone 1 Blood 0 Control Atrazine- Control Males Treated Females Males 1. Are the hermaphrodites males with ovaries or females with testes? 2. What happens when they become adults? 4 CON 4 4 ATR 12 10 8 Pairings 6 4 2 Successful 0 Control Atrazine 35 30 (ng/ml) 25 20 15 10 5 0 Plasma testosterone Plasma 4 4 4 4 4 4 100 66 Fertile 33 Percent 0 Control Atrazine- Males Treated Males Control Atrazine 1. Are the hermaphrodites males with ovaries or females with testes? 2. What happens when they become adults? Parents Effects in other species of frogs? DO EFFECTS OCCUR IN THE WILD? 2.9 – 29 million 10,000 1,000 100 (ppb) 10 DRINKING WATER 1 0.1 Intersexes (Hermaphrodites) Atrazine 0.01 0.001 Runoff Temporary Permanent Precipitation Pools Water Atrazine use (kg/km2) Zero or data missing Less than 0.4 0.4 to 2.4 2.5 to 9.2 9.3 to 28.7 More than 28.7 SPRING Atrazine Metolachlor Alachlor Nicosulfuron Methoxyacetyl Propiconazole Tebupirimphos Cyfluthrin Cyhalothrin 1212 10 9 Control 8 66 34 2 00 40 50 60 70 80 90 1010 88 Atz+S-met 66 44 Metamorphosis 22 0 0 40 50 60 70 80 90 10 10 8 MIX 8 6 6 4 4 2 2 Number at Number 0 40 50 60 70 80 90 0 40 60 80 Time (days) 0.6 ppb Atrazine 15.3 ppb Atrazine Control Atrazine + S-met Mix 4000 0 0 (mg) 3000 0 0 Weight 2000 0 0 Body Body 1000 0 0 40 50 60 70 80 400 50 60 70 80 90 100 1100 50 60 70 80 90 100 1 Time to complete tail reabsorption (days) CONTROL MIX pesticides Stress hormone (Corticoids) Immunosuppression, Decreased growth, Retarded development, Inhibited metamorphosis 4 3 (ng/mL) 2 CORT 1 Blood 0 Control Pesticide Males -treated Males temperature desiccation pesticides density StressStresshormonehormone (Corticoids) (Corticoids) PESTICIDESpesticides Santa Margarita Atascadera Salinas 37 35 age 33 t S 31 29 Santa Margarita Atascadera Salinas 120 (mm) 60 Length 0 Margarita Atascadera Salinas Santa Margarita Atascadera Salinas 100 80 total 60 40 20 Percent 0 0 0.0125 0.05 0.125 0.2 0 0.0125 0.05 0.125 0.2 0 0.0125 0.05 0.125 0.2 Yeast injection (g/ml) Salinas, CA 1952 “Salinas was surrounded and penetrated with swamps with tule-filled ponds and every pond spawned thousands of frogs. With the evening the air was so full of their song that it was a kind of roaring silence. It was a veil, a background, and its sudden disappearance as after a clap of thunder was a shocking thing. It is possible if in the night the frog sound should have stopped, everyone in Salinas would have awakened, feeling that there was a great noise. In their millions the frog songs seemed to have a beat and a cadence and perhaps it is the ear’s function to do this just as it is the eyes’ business to make stars twinkle.” (Steinbeck, 1952) 5 4 3 2 1 0 Level 3 Pathogens EEnnvviironronmmentaentall Atmospheric Habitat Invasive PollutantsPollutants Change modif Species Human Disease Nutrition Predation Other Dev. Repro. Level 2 Take Mortality Failure Failure Level 1 ↑Death ↓Recruitment Population Declines FROM TO SILENT NIGHT “In MAMMALS ecoepidemiology, the occurrence of BIRDS an association in more than one REPTILES species and species population is very strong evidence for causation.” (Fox, ref. 10, page 368). AMPHIBIANSAMPHIBIANS FISH MAMMALS BIRDS Pipidae REPTILES Ranidae AMPHIBIANSAMPHIBIANS FISH Bufonidae Hylidae OH OH OH O T O T Aromatase HO E2 AMP PDE CYP19AROM + CREB ATP C C CBP/P3CO P cAMPK + C CREB ADP C P P CREB CREB SF1 CRE SFRE CYP19AROM THE ENDOCRINE DISRUPTOR, ATRAZINE, IMPAIRS REPRODUCTIVE DEVELOPMENT AND FUNCTION IN ALL VERTEBRATE CLASSES EXAMINED Tyrone B. Hayes, Val Beasley, Chris Kennedy, Valerie S. Langlois, Jennifer Rayner, Vance L. Trudeau, Sunny O. Abarikwu, Lloyd L. Anderson, Alison Bleany, Shane de Solla, Olatunde Farombi, WuQiang Fan, Andrew Friedman, Taisen Iguchi, Holly Ingraham, Patrick Kestemont, Jasna Kniewald, Zlatko Kniewald , Radmila Kovacevic, Jane Linn, Enrique H. Luque, Marcello Maggiolini, Krista A. McCoy, Mónica Muñoz-de-Toro, Tomohiro Oka, Cleida A. Oliveira, Frances Orton, Kristina Pogrmic, Sylvia Ruby, Theresa R. Stueve, Miyuki Suzawa, Luz E. Tavera-Mendoza, Hiroshi Ueda, Mutsumi Ueda, Emily Willingham, Anna Bolivar Victor-Costa, Toshihiko Yanase, (three authors redacted) Control Atrazine Fish (Kestemont et al.) Amphibians (Hayes et al.) Reptiles (Muñoz-de-Toro et al.) Birds (Hussain et al.) Mammals (Victor-Costa et al.; Kniewald et al) Atrazine TestTestosteroneosterone SpermSpermproductionproduction Fish Frogs Rats 6 20 15 5 (Ng/mL) (Ng/mL) (pg/ml) 15 12 4 * 9 3 10 testosterone 6 testosterone 2 Testosterone * 5 3 1 * Serum Plasma Plasma 0 0 Control Atrazine- 0 Control Atrazine- Males Treated Males Treated CON ATR Males Males .18 P < 0.009 In humans Swan et al., 2003 .15 (ppb) .12 level .09 .06 Atrazine .03 0 Control Sub-fertile Males Males 240 Lucas et al. 1993 200 (ppb) 160 level 120 80 Atrazine 40 0 Control Sub-fertile Field Males Males Workers (Average) 2400 2000 (ppb) 1600 level 1200 800 Atrazine 400 0 Control Sub-fertile Field Applicators Males Males Workers (Maximum) (Average) Ethnicity of California Agricultural Workers Asian Latinx California: Fifth largest economy in the world 6 1 in 10 jobs are in agriculture 2 1 30% of land