INTERSEX STURGEON IN THE MISSOURI RIVER? - THE EFFECTS OF EMERGING CONTAMINANTS ON A BIG RIVER

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INTERSEX STURGEON IN THE MISSOURI RIVER? - THE EFFECTS OF EMERGING CONTAMINANTS ON A BIG RIVER Shovelnose sturgeon -Scaphirhynchus platorynchus Pallid sturgeon -Scaphirhynchus albus Photo courtesy of Aaron DeLonay

Overview: Our findings Where intersex occurs Historic occurrence Intersex biology Causes Likely chemicals What we (don t) know Photo Courtesy Aaron DeLonay

Sturgeon are Gonochoristic Female Male

Intersex/Hermaphrodite/Testes-ova Eggs Testis Testis Eggs

Intersex in MO River Sturgeon

Normal Early Stage Ovary

Ovarian folds Testis

Microscopic View

Historic occurrence of intersex

Distribution of Intersex Sturgeon % of Intersexed Males 25 20 15 10 5 17 Male photo 18 16 Fix mdc results 19 16 21 Missouri River Mississippi River 14 0 RM 450 St. Joseph RM 330 Napoleon RM 250 Miami RM 298 Saverton RM 200 Franklin Island RM 170 Easley RM 125 Mokane RM 45 Weldon Springs 15 13 15 14 RM 149 Crystal City RM 847 Caruthersville RM 52 Cape Girardeau

Historic Occurrence Percent 12 10 8 6 4 2 of Catch of Males 0 '64-'71 '68-'69 '78-'79 '93-'94 '00-'01 07-'08 Year

So What? Not Normal Reproductive consequences May be increasing Indicative of other problems? Don t know what the cause is Could be related to water/sediment quality

Gametogenesis Primary Germ Cell (PGC) Sperm Eggs

Genetics Hormonal PGC Sperm Eggs

Hypothalmus Pituitary Hormones PGC Hormones Sperm Gonad Eggs

Some Causes of Abnormal Hermaphroditism in Fish Senescence Genetic abnormalities Hybridization Radiation Diet Temperature Environmental disturbances Chemicals

Age and Size Characteristics 1000 800 intersex normal 20 18 16 mm or grams + SE 600 400 14 12 10 8 6 Years + SE 200 4 2 0 FL WT Age 0

Some Proposed Causes of Abnormal Hermaphroditism in Fish Senescence Genetic abnormalities Hybridization Radiation Diet Temperature Environmental disturbances Chemicals

Cl Cl Cl N N H C-Cl 3 DDT AN ENDOCRINE DISRUPTOR IS: C 2 H 3 N H N Atrazine NCH(CH 3 ) 2 H A compound that interferes with, or changes, normal endocrine (hormone) function. Cl OH Cl Cl C CH HO HO Hydroxy PCBs HO Bisphenol A OH 17 alpha-ethinyl Estradiol OH (C 4 H 9 ) (C 4 H 9 ) Sn (C 4 H 9 ) nonylphenol Tributyl Tin

Reproductive Characteristics, Functions, Processes that Can be Altered with Endocrine Disrupting Chemicals: Sexual differentiation and development Offspring sex ratio Gonad and duct development Secondary sexual characteristics Fertility Fecundity Mating and sexual behavior Gametogenesis Hormone levels

WHERE DO EDCS COME FROM? Personal care products Commercial products Detergents Industrial discharge Residential wastewater Municipal WWTP Agriculture

and where do go? 223

620

192

183

MAPS Courtesy of Gust Annis, TNC 9

From forthcoming report on Contaminants of Concern in the MOR Contaminant Trace Elements Water Exceedances Sediment Exceedances Tissue Exceedances Preliminary Risk Conclusion Arsenic 0 of 302 1 of 21 No Data COC Cadmium 6 of 151 2 of 21 0 of 43 COC Chromium 2 of 151 0 of 20 No Data COC Copper 5 of 151 1 of 25 No Data COC Lead 36 of 151 2 of 26 0 of 41 COC Mercury 0 of 90 0 of 15 25 of 43 COC Nickel 0 of 151 5 of 27 No Data COC Selenium 31 of 302 No Data 1 of 2 COC Zinc 3 of 151 1 of 28 No Data COC Nutrients Ammonia 6 of 268 No Data No Data COC Total Nitrogen 320 of 320 No Data No Data COC Total Phosphorous 346 of 346 No Data No Data COC

Pesticides Water Sediment Tissue 2,4-D 0 of 1 No Data No Data DG Carbaryl 1 of 110 No Data No Data DG Chlordane No Data 1 of 4 29 of 141 COC Chlorpyrifos 0 of 206 0 of 4 No Data DG Diazinon 0 of 206 0 of 1 No Data DG Dieldrin 0 of 205 0 of 4 14 of 140 COC DDT No Data 0 of 4 21 of 141 COC DDD No Data 0 of 4 0 of 139 NL DDE 0 of 202 4 of 4 26 of 139 COC Metolachlor 0 of 206 No Data No Data DG Triazine Herbicides 73 of 206 No Data No Data COC Industrial Organics Dioxins/Furans No Data No Data No Data DG Nonylphenol No Data No Data No Data DG PAHs No Data 0 of 4 No Data DG PBDE No Data No Data No Benchmark (94 samples) DG PCBs No Data 4 of 4 133 of 141 COC Other Chlorophyl A 198 of 198 No Data No Data COC Estrogens No Data No Data No Data DG ph 0 of 361 No Data No Data NL

The Missouri Department of Health and Senior Services (DHSS) recommends that all consumers be aware of the positive benefits of eating fish and the potential adverse health effects of contaminants that may be found in fish.

PCBs, Chlordanes 1800 1600 Egg Concentrations 1400 ng/g egg 1200 1000 800 600 400 200 0 2500 2000 Fillet concentrations Year ng/g fillet 1500 1000 Photo courtesy of Aaron DeLonay 500 Data: Courtesy Mike McKee, MO Dept. of Conservation 0 '84 '86 '88 '96 '04 '08 Year

Unlikely Legacy Chemicals: 100000 Total PCBs 14000 Organochlorine Pesticides ng/g Lipid 80000 60000 40000 20000 ng/g Lipid 12000 10000 8000 6000 4000 2000 0 Normal Intersex Normal Intersex Gonad Brain 0-2000 Normal Intersex Normal Intersex Gonad Brain

Flame Retardants: PBDEs 10000 8000 ng/g Lipid 6000 4000 2000 0-2000 Normal Intersex Normal Intersex Gonad Brain

Chemical Suspects? Ethinyl estradiol Estradiol Atrazine Nonylphenols Mercury Heavy metals

How do we link exposure to MOR EDCs to intersex in sturgeon? Eggs Testis A small incision in the belly of a pallid sturgeon to implant a transmitter revealed it was intersex

Must Consider: Exposure Uptake Metabolism Excretion Mechanism and Site of action

Vulnerability of Sturgeon Benthic feeders/dwellers Long-lived Age at first maturity High lipid content Spawning frequency Other?

DEPARTMENT OF THE INTERIOR U.S. FISH AND WILDLIFE SERVICE REGION #6 FY11 ENVIRONMENTAL CONTAMINANTS PROGRAM OFF-REFUGE INVESTIGATIONS SUB-ACTIVITY Evaluation of Endocrine Active Chemicals and Synthetic Hormones in Shovelnose Sturgeon (Scaphirhynchus platorynchus): Implications for the reproduction and recovery of an endangered species

Testicular Malformations

Reduced Size of Testes Testicular Gonadosomatic Index 10 8 6 4 2 0 R2=0.57-2 1000 10000 100000 ng OCPs/g Lipid

Egg Parasites

Teratomas

Embryological Survival (2-3 dpf) (From Buckler, J. Thesis 2011) 120 2 dpf (p<0.01) Survival (cumulative %) 100 80 60 40 20 0 R² = 0.30 0 100 200 300 400 500 600 dl-pcb (ng/g lipid)

Intersex in other MOR species? Egg Sperm

Environmental Stressors Over harvest Channelization Dams Loss of habitat Contaminants? Photo courtesy of Robb Jacobson

Mean Vitellogenin (ng/ml) ± SD 2000 1800 1600 1400 1200 1000 800 600 400 200 2 1 2 2 1 14 0 Intersex Male Immature Females Mature Females

Mimic endogenous hormone Antagonize hormone effects Disrupt hormone synthesis/metabolism Disrupt receptor synthesis/metabolism/function Plasma Binding Protein Estrogen or Mimic Target Cell Protein Synthesis Altered Functional Response Nucleus DNA Receptor mrna Hormone-Receptor Complex