IDENTITIES AND ECOLOGICAL EFFECTS OF ECOESTROGENS PRESENT IN THE TIDAL CALOOSAHATCHEE RIVER

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IDENTITIES AND ECOLOGICAL EFFECTS OF ECOESTROGENS
PRESENT IN THE TIDAL CALOOSAHATCHEE RIVER

• Mariah Arnold New College of Florida
• Dr. Jim Gelsleichter Mote Marine Laboratory,
  Center for Shark Research
• Nancy Szabo Analytical Toxicology Core
  Laboratory, University of Florida

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Wastewater Effluent

• Wastewater (WW) effluent impact on coastal and
  riverine ecosystems has been documented in the
  U.S. and abroad
• WW effluent can contain household chemical
  products, pharmaceuticals, heavy metals,
  hormones, etc.

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Wastewater and Estrogens
Kolpin et al., 2002
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Ethynylestradiol Recycling
Schultz et al., 2000
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Estrogens in the Lab and Field

• Ethynylestradiol is one of the few pollutants for
  which scientists are able to link laboratory
  results with field data
• Nash et al. (2004) showed multi-generational
  effects of ethynylestradiol exposure
• Kidd et al. (2007) saw similar results in wild
  populations exposed to environmentally relevant
  concentrations of ethynylestradiol

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Estrogens in the Caloosahatchee River

• The Caloosahatchee River is a 60 km long and
  connects Lake Okeechobee to the Caloosahatchee
  Estuary (sfwmd.gov)
• Receives WW effluent and surges of
  anthropogenically modified fresh water
• There is previous evidence of estrogenic activity
  in the river (unpublished data)

Caloosahatchee River
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Goals

• Determine the concentrations of specific
  estrogens
• Used an animal model to determine if the
  estrogens were at concentrations that would
  affect wildlife

Hogchokers (Trinectes maculatus)
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Methods

• Vitellogenin (VTG)
• Egg yolk precursor phosphoprotein
• Normally expressed only in mature females but can
  be induced in males or juveniles if they are
  exposed to estrogen
• Detected blood plasma VTG using Western blot
  immunostaining
• Any samples confirmed positive for Vtg were
  quantified by an enzyme-linked immunosorbent
  assay (ELISA)

• Hogchokers (Trinectes maculatus) were our model
  species
• Collected by bottom trawl
• Primary collection site near WW outfall

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Vitellogenin
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Histology

• It is impossible to sex hogchokers using external
  examination
• Routine paraffin histology used to make thin
  sections of gonads for analysis

A
B
Hogchoker testis (A) and immature ova (B)
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Histopathology

• Liver sections were also analyzed to look for
  possible histopathology
• Irregular, tear-drop shaped white spots on
  external surface of livers

Normal hogchoker liver tissue
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Vitellogenin Results
M
C
Mature ova from female Myakka hogchoker
VTG 72.7-133.8 ug/mL
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Histopathology Results

• Histopathology data indicates the Caloosahatchee
  River hogchokers are stressed
• 67 hogchokers with white spots on liver had
  foci of cellular alteration
• The observed foci of cellular alteration appear
  to represent areas of parasitic infection

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Histopathology Results Continued
A
B
C
D
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Other Observations

• Examined hogchoker livers for presence of other
  biomarkers for environmental chemicals
• PAHs and CYP1-A1
• No indication of induction
• Metallothionein (Mt)
• No evidence in hogchokers

Control
CYP1-A1
Mt
BNF
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Conclusions and Summary

• No evidence of estrogenic contamination in
  Caloosahatchee River
• Liver histopathology data indicates that the
  Caloosahatchee River hogchokers are stressed
• We are continuing the attempt to identify the
  exact type and cause of the liver foci of
  cellular alteration
• Tentatively identified as parasitic infection

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Acknowledgements

• Charlotte Harbor National Estuary Program
• NSF REU Grant
• Mote Marine Laboratory Staff
• University of Florida

CHNEP Grant
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