CERP: F2 Experiment

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CERP F2 Experiment

• Focused on exposing adult mummichogs to
• 4 levels of contaminated sediments
• Clean and contaminated food

njwrri.rutgers.edu/graduate/
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Objectives I. F2 experiment

• Mortality estimates
• Measure growth in length and weight
• Quantify egg production
• II. Egg performance experiment (EP)
• Measure egg size, hatching success,
  and size at hatch for larvae

ehpnet1.niehs.nih.gov/docs/ 1996/104-9/innov.html
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F2 Experiment

• Sediment treatment
• Elizabeth River mud diluted to 6.25 (ERL) ,
  12.5 (ERM), and 25 (ERH)
• Fishing Bay (FB) mud as control
• Same as F1 experiment
• Clean/contaminated food treatment
• Contaminated clams Live clams were placed out in
  the ER river on 2 occasions for 6-7 weeks (T)
• Clean clams live clams were bought from a local
  seafood market (Chester and Ware Rivers) (C)
• Clams were mixed with fish gel and water to make
  clam jello
• Example
• ERMT Elizabeth River 12.5 (medium) dilution
  and toxic food treatment

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Characteristics of clean contaminated food
PAH Accumulation in Mercenaria (note scale
change)
Control
Elizabeth River
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F2 Experiment

• 24 50-gallon flow-through
• tanks fitted with manifolds that
• released incoming flow just
• above the sediment surface
• to mix water column
• 3 replicates per treatment
• Extra aeration was added to each tank
• 56 fish were added to each tank ranging from
  40-73 mm (F1)
• Fish were fed 0.03 g/g/day of their body weight
  for the first
• 30 days then switched to 0.06 g/g/day after
  day 30 (same as
• low food from F1)

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F2 Experiment

• 4 sampling days Days 0, 30, 59, and 74
• Experiment ran from 6/11/03- 8/22/03
• Sub-samples of fish removed for biomarker
• expression, chemical analysis, bioenergetic
• partitioning, and growth
• Eggs collected from each tank
• Egg production (daily), hatchability (weekly),
  and egg/larvae size estimates (weekly)

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F2 Experiment

• Fish were fed twice a day
• (by hand)
• Salinity, temperature, DO
• measurements taken daily
• Egg collectors and flow
• rates checked daily
• Tanks were checked daily
• for dead/dying fish

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F2 Experiment
Environmental Parameters
Salinity 7.5 11.1 ppt 2002 11.3-20.9
ppt Temperature 20 24 oC 2002 18.3-22.4
oC DO - 5.1-8.4 mg L-1 - No problems
with hypoxia or anoxic mud this
time! Flow rate 300 mL min 1 - same as
higher flow in 2002
www.elizabethriver.org/monitoring/
mummichogfacts.htm
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Day 59
Plt0.0001
Sediment
B B B B A
A A A
Adult Fundulus mortality by treatment
Food
Sediment
Day 74
C C B B A
A AB AB
A B A B A
B A B
Plt0.0001
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Mortality comparisons from the last day of
sampling for F1 and F2 experiments
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Differences between F1 and F2 experiment
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Adult Fundulus mean length by treatment
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Adult Fundulus mean absolute growth rate in
length (/- SD) for day 59 and 74
No sediment or food effect on growth
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Mean growth on the last day of sampling for F1
and F2 experiment
Slightly slower growth in 2003 than 2002
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Mean egg production by week throughout the F2
experiment
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Mean egg production for F1 and F2 experiments
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Total egg production (/- SD) from F2 experiment
Food effect p0.0096 Sediment effect p0.0527
B A B A B
A B A
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Total egg production for F1 and F2 experiments
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Egg Performance (EP) experiment

• Eggs were collected daily from collectors
• in each tank. Eggs were subsequently
• grouped by week of the F2 experiment
• Eggs incubated for 14 days in standard
• culture media, exposed to air for 2 hours
• and then placed into highly aerated seawater for
  30 minutes
• This hatching process occurred for 3 days,
  eggs unhatched on the 3rd day of the hatching
  process were termed unhatched or dead

www.miljolare.no/virtue/img/Species/
pages/Fish20eggs.php
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EP experiment Mean area (/- SD) of eggs by
week collected from F2 experiment
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EP experiment Mean length (/- SD) of larvae
at hatch by week
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EP experiment Mean hatching success (/- SD)
of eggs by week collected from F2 experiment
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• Egg and larval traits from EP experiments
• F2 showed similar results with F1 that there
• appears to be no difference among sediment
• or food treatments
• Larval length
• Egg area
• Hatching success

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Conclusions

• Growth appeared to be affected by the lower
  food level
• at the start of the expt and the presence of
  disease
• Egg production
• Higher and longer time of egg production than
  F1
• Egg production was significantly higher in the
  toxic tanks
• Dose response curve with higher total egg
  production
• in the FB tanks (marginally significant)
• There were no apparent differences among
  sediment or
• food treatments with larval and egg
• size or traits

www.whoi.edu/fishtox/ bonyfish.html