Citation:

PANGAEA.957629

Name: Seawater carbonate chemistry and larval viability, size, development, and shell mineralization of the eastern oyster (Crassostrea virginica)
Published: 2023-04-19
License: https://creativecommons.org/licenses/by/4.0/
Keywords: Animalia; Bottles or small containers/Aquaria (<20 L); Crassostrea virginica; Development; Gene expression (incl. proteomics); Growth/Morphology; Laboratory experiment; Laboratory strains; Mollusca; Mortality/Survival; Not applicable; Other studied parameter or process; Pelagos; Single species; Zooplankton

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Abstract:

Calcifying marine organisms, including the eastern oyster (Crassostrea virginica), are vulnerable to ocean acidification (OA) because it is more difficult to precipitate calcium carbonate (CaCO3). Previous investigations of the molecular mechanisms associated with resilience to OA in C. virginica demonstrated significant differences in single nucleotide polymorphism and gene expression profiles among oysters reared under ambient and OA conditions. Converged evidence generated by both of these approaches highlighted the role of genes related to biomineralization, including perlucins. Here, gene silencing via RNA interference (RNAi) was used to evaluate the protective role of a perlucin gene under OA stress. Larvae were exposed to short dicer-substrate small interfering RNA (DsiRNA-perlucin) to silence the target gene or to one of two control treatments (control DsiRNA or seawater) before cultivation under OA (pH ~7.3) or ambient (pH ~8.2) conditions. Two transfection experiments were performed in parallel, one during fertilization and one during early larval development (6 h post-fertilization), before larval viability, size, development, and shell mineralization were monitored. Silenced oysters under acidification stress were the smallest, had shell abnormalities, and had significantly reduced shell mineralization, thereby suggesting that perlucin significantly helps larvae mitigate the effects of OA.

Keyword(s):

Animalia; Bottles or small containers/Aquaria (<20 L); Crassostrea virginica; Development; Gene expression (incl. proteomics); Growth/Morphology; Laboratory experiment; Laboratory strains; Mollusca; Mortality/Survival; Not applicable; Other studied parameter or process; Pelagos; Single species; Zooplankton

Supplement to:

Schwaner, Caroline; Espinosa, Emmanuelle Pales; Allam, Bassem (2023): RNAi Silencing of the Biomineralization Gene Perlucin Impairs Oyster Ability to Cope with Ocean Acidification. International Journal of Molecular Sciences, 24(4), 3661, https://doi.org/10.3390/ijms24043661

Documentation:

Gattuso, Jean-Pierre; Epitalon, Jean-Marie; Lavigne, Héloïse; Orr, James; Gentili, Bernard; Hagens, Mathilde; Hofmann, Andreas; Mueller, Jens-Daniel; Proye, Aurélien; Rae, James; Soetaert, Karline (2022): seacarb: seawater carbonate chemistry with R. R package version 3.3.1. https://cran.r-project.org/web/packages/seacarb/index.html

Project(s):

Ocean Acidification International Coordination Centre (OA-ICC)

Comment:

In order to allow full comparability with other ocean acidification data sets, the R package seacarb (Gattuso et al, 2022) was used to compute a complete and consistent set of carbonate system variables, as described by Nisumaa et al. (2010). In this dataset the original values were archived in addition with the recalculated parameters (see related PI). The date of carbonate chemistry calculation by seacarb is 2023-04-18.

Parameter(s):

#	Name	Short Name	Unit	Principal Investigator	Method/Device	Comment
1	Type of study	Study type		Schwaner, Caroline
2	Species, unique identification	Species UID		Schwaner, Caroline
3	Species, unique identification (URI)	Species UID (URI)		Schwaner, Caroline
4	Species, unique identification (Semantic URI)	Species UID (Semantic URI)		Schwaner, Caroline
5	Experiment	Exp		Schwaner, Caroline
6	Treatment	Treat		Schwaner, Caroline
7	Treatment: pH	T:pH		Schwaner, Caroline
8	Gene expression, fold change, relative	Gene expression		Schwaner, Caroline		perlucin
9	Gene expression, fold change, relative, standard deviation	Gene expression std dev	±	Schwaner, Caroline		perlucin
10	Viability	Viab	%	Schwaner, Caroline
11	Viability, standard deviation	Viab std dev	±	Schwaner, Caroline
12	Larval deformity rate	Larval def rate	%	Schwaner, Caroline
13	Larval deformity rate, standard deviation	Larval def rate std dev	±	Schwaner, Caroline
14	Length	l	µm	Schwaner, Caroline
15	Length, standard deviation	l std dev	±	Schwaner, Caroline
16	Day of experiment	DOE	day	Schwaner, Caroline
17	Birefringence intensity	Birefringence I	pixel	Schwaner, Caroline
18	Birefringence intensity, standard deviation	Birefringence I std dev	±	Schwaner, Caroline
19	pH	pH		Schwaner, Caroline		Total scale
20	pH, standard deviation	pH std dev	±	Schwaner, Caroline
21	Partial pressure of carbon dioxide (water) at sea surface temperature (wet air)	pCO2water_SST_wet	µatm	Schwaner, Caroline
22	Partial pressure of carbon dioxide, standard deviation	pCO2 std dev	±	Schwaner, Caroline
23	Calcite saturation state	Omega Cal		Schwaner, Caroline
24	Calcite saturation state, standard deviation	Omega Cal std dev	±	Schwaner, Caroline
25	Aragonite saturation state	Omega Arg		Schwaner, Caroline
26	Aragonite saturation state, standard deviation	Omega Arg std dev	±	Schwaner, Caroline
27	Carbon, inorganic, dissolved	DIC	µmol/kg	Schwaner, Caroline
28	Carbon, inorganic, dissolved, standard deviation	DIC std dev	±	Schwaner, Caroline
29	Carbonate ion	[CO3]2-	µmol/kg	Schwaner, Caroline
30	Carbonate ion, standard deviation	[CO3]2- std dev	±	Schwaner, Caroline
31	Alkalinity, total	AT	µmol/kg	Schwaner, Caroline
32	Alkalinity, total, standard deviation	AT std dev	±	Schwaner, Caroline
33	Temperature, water	Temp	°C	Schwaner, Caroline
34	Salinity	Sal		Schwaner, Caroline
35	Carbonate system computation flag	CSC flag		Yang, Yan	Calculated using seacarb after Nisumaa et al. (2010)
36	pH	pH		Yang, Yan	Calculated using seacarb after Nisumaa et al. (2010)	Total scale
37	Carbon dioxide	CO2	µmol/kg	Yang, Yan	Calculated using seacarb after Nisumaa et al. (2010)
38	Fugacity of carbon dioxide (water) at sea surface temperature (wet air)	fCO2water_SST_wet	µatm	Yang, Yan	Calculated using seacarb after Nisumaa et al. (2010)
39	Partial pressure of carbon dioxide (water) at sea surface temperature (wet air)	pCO2water_SST_wet	µatm	Yang, Yan	Calculated using seacarb after Nisumaa et al. (2010)
40	Bicarbonate ion	[HCO3]-	µmol/kg	Yang, Yan	Calculated using seacarb after Nisumaa et al. (2010)
41	Carbonate ion	[CO3]2-	µmol/kg	Yang, Yan	Calculated using seacarb after Nisumaa et al. (2010)
42	Aragonite saturation state	Omega Arg		Yang, Yan	Calculated using seacarb after Nisumaa et al. (2010)
43	Calcite saturation state	Omega Cal		Yang, Yan	Calculated using seacarb after Nisumaa et al. (2010)

License:

Creative Commons Attribution 4.0 International (CC-BY-4.0)

Status:

Curation Level: Enhanced curation (CurationLevelC)

Size:

1410 data points

Data

Download dataset as tab-delimited text — use the following character encoding:

1 Study type	2 Species UID	3 Species UID (URI)	4 Species UID (Semantic URI)	5 Exp	6 Treat	7 T:pH	8 Gene expression (perlucin)	9 Gene expression std dev [±] (perlucin)	10 Viab [%]	11 Viab std dev [±]	12 Larval def rate [%]	13 Larval def rate std dev [±]	14 l [µm]	15 l std dev [±]	16 DOE [day]	17 Birefringence I [pixel]	18 Birefringence I std dev [±]	19 pH (Total scale)	20 pH std dev [±]	21 pCO2water_SST_wet [µatm]	22 pCO2 std dev [±]	23 Omega Cal	24 Omega Cal std dev [±]	25 Omega Arg	26 Omega Arg std dev [±]	27 DIC [µmol/kg]	28 DIC std dev [±]	29 [CO3]2- [µmol/kg]	30 [CO3]2- std dev [±]	31 AT [µmol/kg]	32 AT std dev [±]	33 Temp [°C]	34 Sal	35 CSC flag	36 pH (Total scale)	37 CO2 [µmol/kg]	38 fCO2water_SST_wet [µatm]	39 pCO2water_SST_wet [µatm]	40 [HCO3]- [µmol/kg]	41 [CO3]2- [µmol/kg]	42 Omega Arg	43 Omega Cal
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Seawater	8.2	0.02	-0.45										8.2	0.01	402	0.05	4.2	0.10	2.7	0.10	1909	38	161	3.0	2126	39	25.20	25	15	8.05	11.98	402.69	403.98	1732.19	164.82	2.74	4.29
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Negative Control	8.2	0.37	0.41										8.2	0.00	407	4.00	4.2	0.03	2.7	0.02	1914	19	160	1.0	2131	19	24.60	25	15	8.06	11.93	394.80	396.07	1737.27	164.80	2.73	4.28
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Gene Silencing	8.2	-1.05	-1.24										8.2	0.01	411	12.00	4.1	0.10	2.6	0.10	1927	35	158	5.0	2141	40	24.67	25	15	8.05	12.27	406.71	408.02	1751.62	163.12	2.70	4.24
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Seawater	7.3	0.75	0.94										7.3	0.03	3508	151.00	0.6	0.10	0.4	0.04	2062	82	22	2.0	1987	87	24.50	25	15	7.15	107.07	3534.04	3545.47	1932.26	22.68	0.38	0.59
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Negative Control	7.3	0.54	0.52										7.3	0.02	3335	229.00	0.6	0.01	0.4	0.01	2081	80	24	0.4	2013	72	24.33	25	15	7.18	102.17	3357.64	3368.51	1954.58	24.24	0.40	0.63
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Gene Silencing	7.3	-0.77	-1.25										7.3	0.04	341	338.00	0.6	1.00	0.4	0.04	2088	64	24	2.0	2018	65	24.76	25	15	7.17	103.96	3454.78	3465.91	1959.90	24.13	0.40	0.63
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Seawater	8.2	0.20	0.05										8.2	0.02	380	11.00	4.4	0.20	2.8	0.10	1936	48	170	9.0	2167	58	24.67	25	15	8.08	11.38	377.39	378.60	1749.29	175.32	2.91	4.56
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Negative Control	8.2	0.13	-0.28										8.2	0.01	368	3.00	4.3	0.20	2.8	0.10	1891	32	160	1.0	2131	19	24.60	25	15	8.11	10.45	345.73	346.85	1700.29	180.26	2.99	4.68
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Gene Silencing	8.2	-0.51	-0.34										8.2	0.00	386	2.00	4.1	0.02	2.6	0.01	1867	9	159	1.0	2084	10	25.00	25	15	8.06	11.49	384.09	385.32	1692.04	163.48	2.71	4.25
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Seawater	7.3	0.56	0.49										7.3	0.10	3861	436.00	0.6	0.10	0.4	0.03	2149	33	22	2.0	2062	23	24.43	25	15	7.13	118.05	3889.60	3902.18	2008.75	22.20	0.37	0.58
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Negative Control	7.3	0.30	0.44										7.3	0.02	3571	120.00	0.6	0.04	0.4	0.02	2164	15	24	1.0	2088	121	24.40	25	15	7.17	109.93	3618.98	3630.69	2029.74	24.33	0.40	0.63
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Gene Silencing	7.3	-1.55	-1.83										7.3	0.01	3708	239.00	0.5	0.02	0.3	0.01	2017	97	20	1.0	1934	90	24.83	25	15	7.12	112.41	3742.09	3754.14	1883.94	20.65	0.34	0.54
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Gene Silencing	8.2			75	11.0	70.700	9.100	54.099	2.251				8.2	0.01	411	12.00	4.1	0.10	2.6	0.10	1927	35	158	5.0	2141	40	24.67	25	15	8.05	12.27	406.71	408.02	1751.62	163.12	2.70	4.24
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Gene Silencing	7.3			70	10.0	92.300	5.500	44.368	3.310				7.3	0.04	341	338.00	0.6	1.00	0.4	0.04	2088	64	24	2.0	2018	65	24.76	25	15	7.17	103.96	3454.78	3465.91	1959.90	24.13	0.40	0.63
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Negative Control	8.2			52	3.0	17.000	8.000	62.623	1.960				8.2	0.00	407	4.00	4.2	0.03	2.7	0.02	1914	19	160	1.0	2131	19	24.60	25	15	8.06	11.93	394.80	396.07	1737.27	164.80	2.73	4.28
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Negative Control	7.3			72	21.0	58.700	4.900	79.980	1.960				7.3	0.02	3335	229.00	0.6	0.01	0.4	0.01	2081	80	24	0.4	2013	72	24.33	25	15	7.18	102.17	3357.64	3368.51	1954.58	24.24	0.40	0.63
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Seawater	8.2			63	19.0	0.000	0.000	93.829	3.800				8.2	0.01	402	0.05	4.2	0.10	2.7	0.10	1909	38	161	3.0	2126	39	25.20	25	15	8.05	11.98	402.69	403.98	1732.19	164.82	2.74	4.29
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Seawater	7.3			73	15.0	54.700	10.200	71.095	2.170				7.3	0.03	3508	151.00	0.6	0.10	0.4	0.04	2062	82	22	2.0	1987	87	24.50	25	15	7.15	107.07	3534.04	3545.47	1932.26	22.68	0.38	0.59
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Gene Silencing	8.2			85	2.0	19.667	8.505	90.291	2.754				8.2	0.00	386	2.00	4.1	0.02	2.6	0.01	1867	9	159	1.0	2084	10	25.00	25	15	8.06	11.49	384.09	385.32	1692.04	163.48	2.71	4.25
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Gene Silencing	7.3			85	2.0	28.000	10.149	85.844	4.583				7.3	0.01	3708	239.00	0.5	0.02	0.3	0.01	2017	97	20	1.0	1934	90	24.83	25	15	7.12	112.41	3742.09	3754.14	1883.94	20.65	0.34	0.54
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Negative Control	7.3			85	4.0	7.000	12.124	90.587	1.422				7.3	0.02	3571	120.00	0.6	0.04	0.4	0.02	2164	15	24	1.0	2088	121	24.40	25	15	7.17	109.93	3618.98	3630.69	2029.74	24.33	0.40	0.63
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Negative Control	8.2			89	0.8	17.333	10.408	96.565	4.550				8.2	0.01	368	3.00	4.3	0.20	2.8	0.10	1891	32	160	1.0	2131	19	24.60	25	15	8.11	10.45	345.73	346.85	1700.29	180.26	2.99	4.68
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Seawater	8.2			73	1.3	0.000	0.000	97.370	3.844				8.2	0.02	380	11.00	4.4	0.20	2.8	0.10	1936	48	170	9.0	2167	58	24.67	25	15	8.08	11.38	377.39	378.60	1749.29	175.32	2.91	4.56
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Seawater	7.3			75	6.0	0.000	0.000	89.845	0.498				7.3	0.10	3861	436.00	0.6	0.10	0.4	0.03	2149	33	22	2.0	2062	23	24.43	25	15	7.13	118.05	3889.60	3902.18	2008.75	22.20	0.37	0.58
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Gene Silencing	8.2									3	61.500	4.600	8.2	0.00	386	2.00	4.1	0.02	2.6	0.01	1867	9	159	1.0	2084	10	25.00	25	15	8.06	11.49	384.09	385.32	1692.04	163.48	2.71	4.25
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Gene Silencing	7.3									3	43.700	12.400	7.3	0.01	3708	239.00	0.5	0.02	0.3	0.01	2017	97	20	1.0	1934	90	24.83	25	15	7.12	112.41	3742.09	3754.14	1883.94	20.65	0.34	0.54
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Negative Control	7.3									3	62.500	0.630	7.3	0.02	3571	120.00	0.6	0.04	0.4	0.02	2164	15	24	1.0	2088	121	24.40	25	15	7.17	109.93	3618.98	3630.69	2029.74	24.33	0.40	0.63
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Negative Control	8.2									3	75.000	5.000	8.2	0.01	368	3.00	4.3	0.20	2.8	0.10	1891	32	160	1.0	2131	19	24.60	25	15	8.11	10.45	345.73	346.85	1700.29	180.26	2.99	4.68
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Seawater	8.2									3	79.800	8.500	8.2	0.02	380	11.00	4.4	0.20	2.8	0.10	1936	48	170	9.0	2167	58	24.67	25	15	8.08	11.38	377.39	378.60	1749.29	175.32	2.91	4.56
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Seawater	7.3									3	61.700	3.500	7.3	0.10	3861	436.00	0.6	0.10	0.4	0.03	2149	33	22	2.0	2062	23	24.43	25	15	7.13	118.05	3889.60	3902.18	2008.75	22.20	0.37	0.58
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Gene Silencing	8.2									5	57.455	1.150	8.2	0.00	386	2.00	4.1	0.02	2.6	0.01	1867	9	159	1.0	2084	10	25.00	25	15	8.06	11.49	384.09	385.32	1692.04	163.48	2.71	4.25
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Gene Silencing	7.3									5	51.489	1.760	7.3	0.01	3708	239.00	0.5	0.02	0.3	0.01	2017	97	20	1.0	1934	90	24.83	25	15	7.12	112.41	3742.09	3754.14	1883.94	20.65	0.34	0.54
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Negative Control	7.3									5	63.415	0.897	7.3	0.02	3571	120.00	0.6	0.04	0.4	0.02	2164	15	24	1.0	2088	121	24.40	25	15	7.17	109.93	3618.98	3630.69	2029.74	24.33	0.40	0.63
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Negative Control	8.2									5	61.442	2.400	8.2	0.01	368	3.00	4.3	0.20	2.8	0.10	1891	32	160	1.0	2131	19	24.60	25	15	8.11	10.45	345.73	346.85	1700.29	180.26	2.99	4.68
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Seawater	8.2									5	70.456	2.190	8.2	0.02	380	11.00	4.4	0.20	2.8	0.10	1936	48	170	9.0	2167	58	24.67	25	15	8.08	11.38	377.39	378.60	1749.29	175.32	2.91	4.56
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	2	Seawater	7.3									5	61.507	1.600	7.3	0.10	3861	436.00	0.6	0.10	0.4	0.03	2149	33	22	2.0	2062	23	24.43	25	15	7.13	118.05	3889.60	3902.18	2008.75	22.20	0.37	0.58
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Gene Silencing	8.2									3	58.885	22.321	8.2	0.01	411	12.00	4.1	0.10	2.6	0.10	1927	35	158	5.0	2141	40	24.67	25	15	8.05	12.27	406.71	408.02	1751.62	163.12	2.70	4.24
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Gene Silencing	7.3									3	28.609	15.403	7.3	0.04	341	338.00	0.6	1.00	0.4	0.04	2088	64	24	2.0	2018	65	24.76	25	15	7.17	103.96	3454.78	3465.91	1959.90	24.13	0.40	0.63
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Negative Control	7.3									3	22.181	6.754	7.3	0.02	3335	229.00	0.6	0.01	0.4	0.01	2081	80	24	0.4	2013	72	24.33	25	15	7.18	102.17	3357.64	3368.51	1954.58	24.24	0.40	0.63
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Negative Control	8.2									3	50.791	10.429	8.2	0.00	407	4.00	4.2	0.03	2.7	0.02	1914	19	160	1.0	2131	19	24.60	25	15	8.06	11.93	394.80	396.07	1737.27	164.80	2.73	4.28
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Seawater	8.2									3	63.289	12.789	8.2	0.01	402	0.05	4.2	0.10	2.7	0.10	1909	38	161	3.0	2126	39	25.20	25	15	8.05	11.98	402.69	403.98	1732.19	164.82	2.74	4.29
laboratory	Crassostrea virginica	marinespecies.org	urn:lsid:marinespecies.org:taxname:140657	1	Seawater	7.3									3	37.429	21.067	7.3	0.03	3508	151.00	0.6	0.10	0.4	0.04	2062	82	22	2.0	1987	87	24.50	25	15	7.15	107.07	3534.04	3545.47	1932.26	22.68	0.38	0.59