oceans acidic and low in oxygen: lessons from estuarine...
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Oceans Acidic and Low in Oxygen: Lessons from Estuarine
Organisms
Lou Burnett & Karen Burnett [email protected], [email protected]
Grice Marine Lab Hollings Marine Lab
Points covered
High CO2 and Low O2 often occur together
Lesson 1: oysters Lesson 2: crustaceans
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http://www.riverlaw.us/fishkills.html
http://thewatchers.adorraeli.com/
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http://www.wlox.com/story/22730148/jubilee-fish-kill-lures-families-to-beach-for-easy-to-catch-seafood
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http://www.oceanservice.noaa.gov/education/kits/estuaries/media/supp_estuar10d_disolvedox.html
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World-wide Oxygen Levels
Diaz & Rosenberg 2008 Science
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Low oxygen, the insidious killer…………….. Nearly always accompanied by elevated CO2.
Oxygen Levels
Lerberg et al. 2000 Estuaries
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Oxygen and CO2
Data provided by Phil Dustan, CofC; published in Burnett 1997 Am. Zool.
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11 redrawn from Melzner et al. 2013 Mar Biol
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Melzner et al. 2013 Mar Biol
13 redrawn from Melzner et al. 2013 Mar Biol
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Dissolved oxygen and pH co-vary
Taken from “Acidification in coastal waters – an example from Waquoit Bay, Massachusetts” Daniel C. McCorkle Woods Hole Oceanographic Institution NAML “Briefing on the Hill” October 2014 http://www.naml.org/policy/briefings.php
Ocean Surface Water
pH CO2 (µatm) preindustrial 8.2 250 present 8.1 390 2100 7.7 – 7.8 700-1000
hypercapnia = ↑CO2
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Coastal South Carolina Water
pH CO2 (µatm) preindustrial 8.2 250 present 8.1 390 2100 7.7 – 7.8 700-1000 present 6.0-8.1 300->20,000
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hypercapnia = ↑CO2
Cochran & Burnett 1996 JEMBE http://cdmo.baruch.sc.edu/
How to understand effects
Habitat Natural history
life cycles habits of organisms
oysters close and are exposed to air crabs and shrimp are active; move and migrate
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Crassostrea virginica
Lesson 1: Oysters
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Dwyer & Burnett 1996 Biol. Bull. Allen & Burnett 2008 JEMBE
That’s CO2 at 20,000 µatm!
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[Ca+2] = 4 mM 11 mM
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Dwyer & Burnett 1996 Biol. Bull. Allen & Burnett 2008 JEMBE
Boyd & Burnett 1999 J. Exp. Biol.
Reactive O2 Intermediate Production in Oyster Hemocytes
+hypercapnia
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Milardo Thesis, CofC
“Dermo” Oyster Disease In vitro growth
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24 Milardo Thesis, CofC
“Dermo” Oyster Disease In vitro growth
Oyster Take-Home Lesson
Being hot and air exposed has some disadvantages.
Intertidal air exposure imparts some protection against “Dermo.”
Some field observations support this.
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A note on larval biology
bivalve larvae are sensitive to both hypercapnia and hypoxia
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Treatment “Ambient” CO2 0.25 torr 0.035 kPa 354 µatm
Treatment “Mid” CO2 0.49 torr 0.066 kPa 651 µatm
Treatment “High” CO2 1.14 torr 0.152 kPa 1,500 µatm
%
Surv
ival
veliger pediveliger metamorphosed
%
Surv
ival
%
Su
rviv
al
hard clam Mercenaria mercenaria
bay scallop Argopecten irradians
Eastern oyster Crassostrea virginica
Days Talmage & Gobler 2009 L&O
Oyster Settlement in Hypoxia
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Baker & Mann 1992 Biol Bull
What about crustaceans?
Atlantic Blue Crab Callinectes sapidus
Pacific Whiteleg Shrimp Litopenaeus vannamei
Lesson 2
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Shrimp Survival
Mikulski et al. 2000 JEMBE 30
Shrimp Survival
Mikulski et al. 2000 JEMBE 31
Hemocytes are abundant
gills of shrimp – Litopenaeus vannamei
10 to 50 x 106 mL-1
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lamellae
Photos Courtesy of Don Lovett
Lamellar Gills
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Gill Lamella
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seawater
seawater
hemolymph
Photo Courtesy of Don Lovett
Gill Lamella
hemocyte aggregation
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seawater
seawater
hemolymph
Elimination of Hemocytes
hemocyte aggregation
Jennifer Ikerd
36 Photo Courtesy of Don Lovett
Immune response has some down sides!
Immune response itself decreases O2 transport hemocytes clog gills ↑ vascular resistance ↓ oxygenation at gills ↓ O2 uptake ↓ exercise performance
Burnett et al. 2006 Biol. Bull. Marissa Roman, REU student 2012
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Immune responses & other functions are affected by hypoxia and hypercapnia!
Boleza et al. 2001 Fish Shellfish Immunol. Burgents et al. 2005 Biol. Bull. Tanner et al. 2006 Comp. Biochem. Physiol. Macey et al. 2008 Fish Shellfish Immunol. Hardy et al. 2013 Am. J. Physiol.
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Shrimp on a Treadmill
David Scholnick Pacific University
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Blue Crab Exercise
Thibodeaux et al. 2009 J. Exp. Biol. 40
Walking Exercise
H2O
Stover et al. 2013 Biol. Bull. 41
Walking Exercise
42 Stover et al. 2013 Biol. Bull.
Fatigue Behaviors in Blue Crabs
20% air sat.
50% air sat.
100% air sat.
43 Stover et al. 2013 Biol. Bull.
Fatigue Behaviors in Blue Crabs
20% air sat.
50% air sat.
100% air sat.
50% air sat. + 2% CO2
44 Stover et al. 2013 Biol. Bull.
Adaptations to Low Oxygen are Muted by CO2
Organisms respond to chronic low oxygen in similar ways. ↑ concentration of respiratory pigment
(e.g., hemoglobin, hemocyanin) ↑ oxygen affinity of pigment ↑ vascularization of tissues
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Effects of Hypoxia on the Atlantic Blue Crab, Callinectes sapidus
25 days at Po2 = 30-35% air saturation
Normoxia Hypoxia
Hemocyanin (g mL-1) 3.11 4.40
Oxygen affinity Increase in affinity due to lactate urate calcium
Increase in affinity due to structural change in
hemocyanin
deFur et al. 1990 Biol. Bull.
Hemocyanin O2 Affinity
Po2 (torr)
%O
2 Sa
tura
tion
100
50
0
• P50 the Po2 required to half saturate Hc with O2
• High affinity in hypoxia = good!
P50 is a measure of O2 affinity
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Hemocyanin O2 Affinity
pH
Log
P50
(to
rr)
high affinity
low affinity
better for hypoxia
not so good for hypoxia
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CO2-specific Effect
P 50
(tor
r)
pH
Mangum & Burnett 1986 Biol. Bull.
Pco2 1.5 torr
Pco2 15 torr
large ↑ in O2 affinity due to CO2
Atlantic blue crab Callinectes sapidus
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Transcriptomic Responses
News Flash… Hypoxia-induced production of hemocyanin is muted by CO2!
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CO2 Turns Off Hemocyanin Synthesis
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Construct Stranded Library
~400 bp
Sequenced Transcriptome (Illumina)
RNAseq
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4 h 24 h
Normoxia
Hypoxia
Hypercapnic Hypoxia
Gene networks
Hemocyanin Regulation
High Throughput RNA Sequencing (RNA-Seq)
Johnson et al., in revision, Physiological Genomics
Lessons from Estuarine Organisms
life in O2 and CO2 environments more extreme and more variable than the open ocean
immune functions are compromised by hypoxia and made worse by hypercapnia
immune response itself reduces performance performance is strained by hypoxia hemocyanin function may be limited by hypercapnia molecular approaches will provide answers to many
questions!!! 52
Support
NSF Karen & Lou Burnett & Summer REU Program
Oyster Disease Research Program (NOAA)
US Dept. of Agriculture
SC Sea Grant
Oceans & Human Health Initiative (NOAA)
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Oceans Acidic and Low in Oxygen: Lessons from Estuarine
Organisms Lou Burnett & Karen Burnett
[email protected], [email protected] http://burnettl.people.cofc.edu
Grice Marine Lab Hollings Marine Lab
Citations Allen, S. M., and L. E. Burnett. 2008. The effects of intertidal air exposure on the respiratory physiology and
the killing activity of hemocytes in the Pacific oyster, Crassostrea gigas (Thunberg). J. Exp. Mar. Biol. Ecol. 357:165-171.
Baker, S. M., and R. Mann. 1992. Effects of hypoxia and anoxia on larval settlement, juvenile growth, and juvenile survival of the oyster, Crassostrea virginica. Biol. Bull. 182:265-269.
Boleza, K. A., L. E. Burnett, and K. G. Burnett. 2001. Hypercapnic hypoxia compromises bactericidal activity of fish anterior kidney cells against opportunistic environmental pathogens. Fish Shellfish Immunol. 11:593-610.
Boyd, J. N., and L. E. Burnett. 1999. Reactive oxygen intermediate production by oyster hemocytes exposed to hypoxia. J. Exp. Biol. 202:3135-3143.
Burgents, J. E., K. G. Burnett, and L. E. Burnett. 2005. Effects of hypoxia and hypercapnic hypoxia on the localization and the elimination of Vibrio campbellii in Litopenaeus vannamei, the Pacific white shrimp. Biol. Bull. 208:159-168.
Burnett, L. E. 1997. The challenges of living in hypoxic and hypercapnic aquatic environments. Am. Zool. 37:633-640.
Burnett, L. E., J. D. Holman, D. D. Jorgensen, J. L. Ikerd, and K. G. Burnett. 2006. Immune defense reduces respiratory fitness in Callinectes sapidus, the Atlantic blue crab. Biol. Bull. 211:50-57.
Cameron, J. N. 1978. Effects of hypercapnia on blood acid-base status, NaCl fluxes, and trans-gill potential in freshwater blue crabs Callinectes sapidus. J. Comp. Physiol. 123:137-141.
Cochran, R. E., and L. E. Burnett. 1996. Respiratory responses of the salt marsh animals, Fundulus heteroclitus, Leiostomus xanthurus, and Palaemonetes pugio to environmental hypoxia and hypercapnia and to the organophosphate pesticide, azinphosmethyl. J. Exp. Mar. Biol. Ecol. 195:125-144.
deFur, P. L., C. P. Mangum, and J. E. Reese. 1990. Respiratory responses of the blue crab Callinectes sapidus to long-term hypoxia. Biol. Bull. 178:46-54.
Diaz, R. J., and R. Rosenberg. 2008. Spreading dead zones and consequences for marine ecosystems. Science 321:926-929.
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Citations Dwyer, J. J., III, and L. E. Burnett. 1996. Acid-base status of the oyster Crassostrea virginica in response
to air exposure and to infections by Perkinsus marinus. Biol. Bull. 190:139-147. Hardy, K. M., K. G. Burnett, and L. E. Burnett. 2013. The effect of hypercapnic hypoxia and bacterial
infection (Vibrio campbellii) on protein synthesis rates in the Pacific whiteleg shrimp, Litopenaeus vannamei. Am. J. Physiol.-Reg. I. 305:R1356-R1366.
Lerberg, S. B., A. F. Holland, and D. M. Sanger. 2000. Responses of tidal creek macrobenthic communities to the effects of watershed development. Estuaries 23:838-853.
Macey, B. M., I. O. Achilihu, K. Burnett, and L. Burnett. 2008. Effects of hypercapnic hypoxia on inactivation and elimination of Vibrio campbellii in the Eastern oyster, Crassostrea virginica. Appl. Environ. Microbiol. 74:6077-6084.
Mangum, C. P., and L. E. Burnett. 1986. The CO2 sensitivity of the hemocyanins and its relationship to Cl- sensitivity. Biol. Bull. 171:248-263.
Melzner, F., J. Thomsen, W. Koeve, A. Oschlies, M. Gutowska, H. Bange, H. Hansen, and A. Körtzinger. 2013. Future ocean acidification will be amplified by hypoxia in coastal habitats. Mar. Biol. 160:1875-1888.
Mikulski, C. M., L. E. Burnett, and K. G. Burnett. 2000. The effects of hypercapnic hypoxia on the survival of shrimp challenged with Vibrio parahaemolyticus. J. Shellfish Res. 19:301-311.
Rathburn, C. K., N. J. Sharp, J. C. Ryan, M. Nealy, M. Cook, R. W. Chapman, L. E. Burnett, and K. G. Burnett. 2013. Transcriptomic responses of juvenile Pacific whiteleg shrimp, Litopenaeus vannamei, to hypoxia and hypercapnic hypoxia. Physiol. Genomics 45:794-807.
Scholnick, D. A., K. G. Burnett, and L. E. Burnett. 2006. Impact of exposure to bacteria on metabolism in the penaeid shrimp Litopenaeus vannamei. Biol. Bull. 211:44-49.
Stover, K. K., K. G. Burnett, E. J. McElroy, and L. E. Burnett. 2013a. Locomotory fatigue and size in the Atlantic blue crab, Callinectes sapidus. Biol. Bull. 224:63-67.
Stover, K. K., K. G. Burnett, E. J. McElroy, and L. E. Burnett. 2013b. Locomotory fatigue during moderate and severe hypoxia and hypercapnia in the Atlantic blue crab, Callinectes sapidus. Biol. Bull. 224:68-78.
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Citations Talmage, S. C., and C. J. Gobler. 2009. The effects of elevated carbon dioxide concentrations on the
metamorphosis, size, and survival of larval hard clams (Mercenaria mercenaria), bay scallops (Argopecten irradians), and Eastern oysters (Crassostrea virginica). Limnol. Oceanogr. 54:2072-2080.
Tanner, C. T., K. G. Burnett, and L. E. Burnett. 2003. Effects of hypoxia and pH on phenoloxidase activity in the blue crab, Callinectes sapidus. Integr. Comp. Biol. 43:1043-1043.
Thibodeaux, L. K., K. G. Burnett, and L. E. Burnett. 2009. Energy metabolism and metabolic depression during exercise in Callinectes sapidus, the Atlantic blue crab: effects of the bacterial pathogen Vibrio campbellii. J. Exp. Biol. 212:3428-3439.
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