Org. Geochem.Vol. 20, No. 5, pp. 563-577, 1993 0146-6380/93$6.00+ 0.

00
Printed in Great Britain.All rights reserved Copyright © 1993PergamonPress Ltd

Seasonal variations in the sources and alteration of organic matter

associated with recently-deposited sediments
ELIZABETHA. CANUEL* and CHRISTOPHERS. MARTENS
Curriculum in Marine Sciences, 12-5 Venable Hall, University of North Carolina at Chapel Hill,
Chapel Hill, NC 27599-3300, U.S.A.

(Received 9 July 1992; accepted in revised form 22 December 1992)

Abstract--Analysis of lipid biomarker compounds associated with surface sediment (04).5 or 0-1.0 cm)
deposited monthly in Cape Lookout Bight (CLB), North Carolina, U.S.A. revealed seasonal trends in the
relative importance of various sources of organic matter. Seasonalityin these sources was reflected through
variations of source-specific biomarkers in three classes of extractable lipid components--fatty acids,
sterols and hydrocarbons, over an 18 month period. Samples collected during periods of sediment
accumulation (winter/spring, generally) showed an increase in the relative abundance of algal-derived
components. Summer months were characterized by negligibleaccumulation of sediment and a threefold
increase in fatty acids of bacterial origin (i.e. odd numbered n-alkanoic acids and iso- and anteiso-
branched acids). Further evidence for the activity of bacteria during summer months was seen by increases
in the 5~(H)-cholestan-3fl-ol to cholest-5-en-3fl-ol ratio. These data indicate that bacterially-mediated
processes incorporate organic matter derived from other sources, and accumulated during other times of
the year, into biomass. As a result, in situ processes control the composition of preserved organic matter
at least as much as temporal changes in the delivery of materials derived from allochthonous sources.

Key words--lipids, biomarker compounds, bacteria, organic matter, sediment-water interface, microbial
repackaging, marine sediments

INTRODUCTION near the sediment-water interface are probably more

Despite the overriding importance of organic matter
production and degradation in biogeochemical cy-
cling processes, many basic questions about the
sources and behavior of organic matter in coastal
environments remain to be answered. Questions re-
garding how the nature of organic matter affects the
rate and extent of degradation and what factors are
important in controlling the reactivity of different
types of organic matter have only recently been
addressed (e.g. Henrichs and Doyle, 1986; Westrich
and Berner, 1984). Other aspects of the carbon cycle
which have received little attention are the relative
importance of local sources of organic matter to
coastal sedimentary processes and the importance of
seasonal fluctuations in the delivery of organic matter
to nearshore systems (Hedges et al., 1988a, b). This
includes understanding how rates of delivery to the
sediment-water interface vary over both short (weeks
to seasons) and long (greater than a year) time scales.
Degradation processes occurring at or near the
sediment-water interface are sensitive to temporal
changes in the reactivity of organic matter being
delivered to nearshore environments. Variability in
the physical environment (e.g. 02 delivery, tempera-
ture) can be important as well. Temporal variations
in the delivery and degradation of organic matter
*To whom all correspondence should be addressed at: U.S.
Geological Survey, Water Resources Division, 345 Mid-
dlefield Road, MS-496, Menlo Park, CA 94025, U.S.A.
significant in coastal systems where productivity is
high and water column residence times are short. As
a result, a large percentage of the organic matter
reaching the sediment-water interface is likely to be
composed of "fresher", more metabolizable constitu-
ents. This availability of labile components results in
the rapid turnover of organic matter with up to 85%
of the total near-bottom organic carbon flux being
utilized at or near the sediment-water interface
(Reimers and Suess, 1983). The rapid recycling of
metabolizable organic matter is responsible for the
return of essential nutrients to the overlying waters.
These processes also control redox conditions which,
in turn, determine the pathways by which organic
matter is metabolized. Temporal variations in or-
ganic matter input and burial rate may thus deter-
mine the extent to which organic constituents are
utilized rather than becoming buried and ultimately
preserved.
Information regarding short-term variations in the
composition of organic matter arriving at the sedi-
ment-water interface is often lost due to the time-
integrating nature of marine sedimentation and mix-
ing processes. This is especially true in the nearshore
environment where the rate of delivery, as well as
rates of degradation by microbially-mediated pro-
cesses can be high. The preferential loss of metaboliz-
able substrates by diagenetic processes leaves behind
a source signal enriched in more refractory com-
ponents and results in a biased depositional record.

563
564 EL]ZAnETHA. CANUELand CHRISTOPHERS. MARr~t~S

This paper traces changes in the distribution of
lipid biomarker compounds in a rapidly-accumulat-
ing coastal sediment over seasonal time-scales in an
attempt to understand temporal changes in the im-
portance of various organic matter sources. Because
the accumulation rate of new particulate matter is
well-constrained by a previous Be-7 radiochrono-
metric study (Canuel et aL, 1990), the effects of
changes in input can be separated from those occur-
ring as a result of the in situ diagenetic alteration of
lipid components. Source-specific compounds from
lipid classes with differing chemical reactivities (hy-
drocarbons, fatty acids and sterols) will be used to
show how short-term variations (i.e. monthly to
seasonal) in the relative abundance of organic matter
derived from different sources are reflected in the
sedimentary record of a nearshore environment.
STUDY SITE
The site of the study was Station A-I in Cape
Lookout Bight (CLB), NC (Fig. 1). This semi-
enclosed marine basin is located approx. 105 km
southwest of Cape Hatteras at the southern end of the
Outer Banks island chain in North Carolina, U.S.A.
Work conducted at this site over the past 15 yr has
resulted in well-established carbon and nutrient bud-
gets (Klump and Martens, 1981, 1987; Martens and

N.C. ,~, • ~"~ /

,~7/ ~&© -,-. ,1

. ~_ .¢" ~
I _ oo,,_,..,,._.s "" / x~
...3;
.~Studysite ~ I~''"~~x~, /'',~ x~ ~ ,~

- N__._ ~ -~ , " .... t _ - 7 w, ~2

.=_~ ,v. ~ , ~.._-.:;//

Atlantic Ocean

N
1.0km
I I

i
l/ Barden
Inlet
*A-1 /

Marshland
Seagrass
habitat

76*35' 76"30'
1

Fig. !. Map of the Cape Lookout Bight, NC site showing the extensive marsh and scagrass habitats
associated with the region. Station A-1 was the location at which our sampling took place.
 Seasonal variations in the sources and alteration of organic matter
Klump, 1984; Martens et al., in press), sediment
accumulation rates (Chanton et al., 1983; Canuel
et al., 1990), organic matter inputs (Haddad and
Martens, 1987) and rates of carbon and nitrogen
remineralization (Crill and Martens, 1987; Burdige
and Martens, 1988).
Local sources of organic matter include highly
productive marsh (Spartina alterniflora) and seagrass
environments ( Zostera marina and Halodule
wrightii), phytoplankton productivity in the offshore
waters and the back-barrier island sounds, eroding
shoreline peats (derived from buried lagoonal marsh
plants and uncovered during barrier island mi-
gration) and terrestrial inputs delivered via the nu-
merous river-estuary systems associated with the
North Carolina coast. Delivery appears to be pre-
dominantly by horizontal transport processes
whereby the Bight acts as a natural "sediment trap"
capturing fine-grained particulate matter, as well as
larger phytoplankton and seagrass debris carried into
the Bight by tidal flushing through Barden Inlet (see
Fig. 1). Vertical flux to the sediment bed appears to
be primarily in the form of large marine snow
aggregates (Wells and Shanks, 1987; Wells, 1988).
Remineralization of organic matter in the Bight's
fine-grained sediments is dominated by anaerobic
processes--sulfate reduction and methanogenesis
(Martens and Klump, 1984). Rates of these processes
are high in response to the availability of organic
matter supplied by the high rates of sediment ac-
cumulation (10.3_+0.7cm yr-l; Chanton et al.,
1983). Rates of remineralization and fluxes of both
oxidants and end-products exhibit pronounced sea-
sonal cycles. This is primarily due to the 20°C average
temperature change occurring over an annual cycle
(Crill and Martens, 1987; Martens and Klump, 1984).
However, conditions approximating annual steady
state have existed at the study site since before 1971.
Evidence for steady state includes annually repro-
ducible pore water nutrient profiles and fluxes, good
agreement between depth-integrated rate measure-
ments of sulfate reduction and sulfate fluxes calcu-
lated from pore water concentration profiles
(Martens and Klump, 1984; Crill and Martens, 1987;
Klump and Martens, 1987) and agreement between
rates of sediment accumulation determined by rela-
tively long- (Pb-210; Chanton et al., 1983) and short-
term (Be-7; Canuel et al., 1990) radiochronometric
tracers.
METHODS
Sample collection
Cores were collected using 50 crn long plastic core
tubes with an outer diameter of 8 crn. All cores
were taken by divers to minimize disturbance of the
flocculent upper 2-3 mm of sediment. Samples were
transported approx. 20 km to the University of N.
Carolina Institute of Marine Sciences (IMS) in ambi-
ent seawater and in darkness. Upon arrival at IMS
565
the cores were sectioned and the uppermost sediment
interval (generally 0.0-0.5 cm) was subsampled for
the analyses described in this study. Further analyses
of underlying deeper sediments in selected cores will
be discussed in a later manuscript. Sediment sections
were placed in previously-combusted (450°C; 4.5 h)
glass jars and sealed with aluminum foil-lined lids.
The samples were placed in a cooler of dry ice and
transported to the University of N. Carolina, Chapel
Hill where they were stored at -80°C.
Organic extraction
Prior to extraction, sediment sections were thawed
and homogenized. Aliquots (,,~ 10 g wet weight) were
placed in combusted jars, and 30 ml methanol added
to each. The solvent-extractable lipids were obtained
by successive extraction of the sediment with CH3OH
(1 h) and CH2C12:CH3OH (2:1, v:v; 3 x 1 h) aided by
sonication. The extractions were conducted at cool
temperatures (~20°C) by adding ice to the sonic
bath. The supernatant from each of the extractions
was combined, washed with a saturated NaC1 sol-
ution and the non-polar phase (CH2C12) was re-
moved. The polar phase was then partitioned into
hexane three times in a separatory funnel. The com-
bined organic (i.e. methylene chloride and hexane)
phases were dried over anhydrous Na2SO4 crystals for
at least overnight. The extract was transferred to
another flask, the Na:SO4 crystals rinsed with hexane
and the extract plus rinse concentrated by rotary
evaporation. The concentrated extract was trans-
ferred to tared vials and the total lipid extract was
weighed after drying under a gentle stream of N2.
Saponification and derivatization
Lipid extracts were dissolved in a known volume of
solvent and an aliquot of the total extract represent-
ing 3--10 mg lipid was transferred to 13 ml centrifuge
tubes. The extract was evaporated to dryness under
a gentle stream of N2. Samples were saponified using
1N KOH in aqueous CH3OH (10% H20) by heating
for I h in a boiling water bath. After cooling, neutral
lipids were extracted using three 5 ml volumes of
hexane. The pH of the saponified extract was then
brought to 2 by the addition of 6N HCI and acidic
lipids were extracted, again using three 5 ml portions
of hexane. Both neutral and acidic lipids were al-
lowed to sit in the presence of anhydrous Na2SO4
crystals overnight in order to remove traces of water.
The acidic lipids were then derivatized to their methyl
ester derivatives using BFa-CHaOH (5%) while
heating at 100°C in a water bath. After cooling, the
methyl esters were extracted into hexane and traces of
water removed by adding Na2SO4 crystals.
Both the neutral and acidic (as fatty acid methyl
esters; FAMEs) lipids were separated into their con-
stituent classes using adsorptive chromatography on
silica gel columns (Bio-Sil A; Bio-Rad; 70-150/~m).
The neutral lipids were separated on 5% deactivated
SiO2 using solvents of increasing polarity from
566 ELIZABETHA. CANUELand CHRISTOPHERS. MARTENS
hexane through 20% ethyl acetate-hexane (Wake-
ham and Canuel, 1990). The fractions containing
hydrocarbons (F1; hexane) and sterols (F5/6; 15 and
20% ethyl acetate in hexane) were collected. The
derivatized acidic lipids were separated on fully acti-
vated SiO2 columns and fractions containing FAMEs
( F 3 / 4 : 5 and 10% ethyl acetate in hexane) were
collected. Fractions collected from the chromato-
graphic separations were concentrated by rotary
evaporation, transferred to vials and stored in a
freezer in a small volume of solvent until analysis by
gas chromatography.
Gas chromatographic analysis
Fatty acid methyl esters (FAMEs), hydrocarbons
and sterols (as TMS ethers) were analyzed on a
30m × 0.32 mm i.d. DB-5 (dimethylphenylsilicone)
fused silica capillary column (J & W Scientific).
Sterols were derivatized to their TMS (trimethylsilyl)
ether derivatives at 80°C using BSTFA [bis(trimethyl-
silyl)trifluoroacetamide] and acetonitrile (Pierce)
prior to analysis by gas chromatography. A Hewlett
Packard 5890 Series II gas chromatograph was used
with injections made in either the on-column or
splitless mode (results for splitless and on-column
injections agreed to within _ 10%). Helium was used
as the carrier gas and detection was by flame ioniz-
ation detector (FID). Temperature programs varied
according to the compound class being analyzed:
100-300°C at 3°C min -l for fatty acid methyl esters,
70-310°C at 4°C min -l for hydrocarbons and
120-315°C at 3°C min i for alcohol/sterol fractions.
Temperatures were held at the upper temperature
until all peaks had eluted.
Data were collected and processed using a micro-
computer and software developed by Waters Chro-
matography (Baseline Program). Response factors
for all components were calculated relative to exter-
nal standards. Quantification was made using in-
ternal standards: 5~(H)-cholestane for the sterols,
hexadecane for hydrocarbons and methyl henei-
cosanoate for the FAMEs, added prior to GC analy-
sis. Identifications were made by comparing relative
retention times with known standard mixtures.
Equivalent chain lengths (ECL) were calculated and
also used for identification purposes. Peak identifi-
cations were verified by G C - M S analysis (Hewlett
Packard 5890A gas chromatograph interfaced with
the 5971 Mass Selective Detector or a Hewlett Pack-
ard 5890 gas chromatograph coupled to a Finnigan
Incos 50 mass spectrometer-data system). Samples
were run in the El (electron impact) mode and spectra
generally collected over the mass range 50-550.
Organic carbon/total nitrogen determination
Samples were prepared for organic carbon and
total nitrogen analyses as follows. Aliquots of the
sediment sample were placed in combusted glass
scintillation vials and freeze-dried. The freeze-dried
samples ( ~ 1 g) were ground to a fine powder with an
agate mortar and pestle and treated with a cold HCi
solution (5% Ultrex HCI in distilled/deionized H20)
until no further release of CO2 was visible (generally
~ 5 m l ) . Acidified samples were sonicated briefly
( ~ 3 0 m i n ) and left in a fume hood overnight.
Samples were again sonicated the following morning
and then freeze-dried. The dry sediments were re-
equilibrated with the atmosphere and weighed to
correct for the addition of H20 to hygroscopic chlor-
ide salts (CaC12 • H20 ). This freeze-drying procedure
minimized losses and results of this method compared
well with previous results (Martens and Klump, 1984;
Martens et al., in press).
Organic carbon and total nitrogen content were
determined using a Carlo Erba NA 1500 elemental
analyzer. Quantitation of nitrogen and carbon was
made using acetanilide as an external standard. The
CO2 resulting from combustion of organic carbon
was collected for subsequent 6 ~3Coc analysis. These
analyses were performed on a Finnigan MAT 251
stable isotope mass spectrometer. All isotopic values
are given in the " 6 " notation in parts per thousand
(%o) deviations and expressed relative to the PDB
standard. Precision of the 613C analyses was
_+0.20%o.
Spatial variability
Two separate studies measured horizontal variabil-
ity in several constituents of interest over 3-28 m 2
areas in CLB. The first spatial variability study took
place in September 1987 and examined differences in
the % organic carbon, total nitrogen contents,
6~3Coc, as well as other constituents (Canuel et al.,
1990). Ten cores (4.5 cm i.d.; 10-15 cm length) were
collected near Station A- 1. A single core was collected
at a central site and three cores spaced approx. 1 m
apart were collected along each of three transects
moving away from the location at which the first core
was collected.
A second variability study was conducted in July
1990 in order to examine spatial variability in only
the lipid constituents. In this study, a single core was
collected near Station A-1 and three additional cores
were collected approx. 1 m away from the central
location. The surface sediment (0-1 cm) was removed
from each of the four cores and placed in individual
glass jars containing solvent (CH2C12: CH3OH; 2 : 1 by
volume) and transported to Chapel Hill on ice. These
samples were processed immediately using the
methods described above except that the initial
CH3OH extraction step was omitted.
RESULTS
Characterization of the Surflcial Sedimentary
Environment
Elemental composition
Percent organic carbon and total nitrogen contents
for the surface sediments ( < 0 . 5 c m ) are given in
 Seasonal variations in the sources and alteration of organic matter 567
Table 1. Cape LookoutBight, NC: surficial sediment (0--0.5crn) characteristics
OrE C Total N Accumulation* Lipid Lipid/OC
Sample (%) (%) C:N (g crn -2) (mE g - ' dry sed) (%)
28 June 87 0.00 1.80
13 Aug 87 0.00 3.30
25 Sept 87 4.00 0.43 9.4 1.39
21 Nov 87 3.40 0.39 8.7 0.00 2.10 6.18
10 Dec 87 3.35 0.40 8.4 0.00 2.33 6.97
27 Feb 88 3.43 0.41 8.1 0.56 4.60 13.40
29 Mar 88 3.15 0.40 8.0 0.96 5.34 16.96
30 Apr 88 2.90 0.34 8.5 0.85 3.10 11.95
19 Jun 88 3.40 0.42 8.2 1.03 4.11 12.07
1 Jul 88 3.40 0.36 9.5 0.00 2.73 8.02
20 Jul 88 3.30 0.38 8.7 0.00 2.90 8.79
17 Aug 88 3.80 0.39 9.8 0.00 5.41 14.24
2 Sept 88 3.40 0.40 8.4 -0.71 3.30 9.71
1 Oct 88 2.80 0.32 8.7 0.97 2.89 10.31
:~ (errorT) 3.4 (0.3) 0.39 (0.03) 8.7 (0.6) 3.4 (1.2) 10.8 (3.3)
*Note that accumulation is for the period of time since the previous core was collected (Canuel et al., 1990).
t E r r o r determined from spatial variability experiment (see Table 2).

Table 1. Monthly values ranged from 2.8 to 4.0% organic carbon and total nitrogen were 5.0 and 6.3%,
organic carbon and 0.32 to 0.43% total nitrogen. The respectively and 1.4% for the 613Coc. Replicate
carbon concentrations were highest during late sum- %OC, TN and fi~3Coc analyses yielded C.V. similar
mer (September 1987 and August 1988). The nitrogen to the spatial variability study indicating that spatial
concentrations, however, did not exhibit any season- variability was not significantly different from the
ality. C : N weight ratios ranged from 8.0 to 9.8 (Table analytical error. Lipid constituents exhibited greater
1) and were highest during September 1987 and spatial variation than was seen in the elemental
August 1988. analyses. The coefficients of variation for the hydro-
carbon, fatty acid and sterol classes were 17, 27 and
Stable carbon isotopes 22%, respectively. Analytical reproducibility for the
The isotopic composition of the total organic lipid methods was only +__10%, less than that seen in
carbon (6~3Coc) displayed a shift of about 2.5%0 the spatial study. Spatial variability studies in com-
during the study period (Fig. 2). Values were heaviest parable environments (fine-grained anoxic sediments)
during late summer (September 1987 and Septem- have yielded variations (spatial + analytical) on the
ber/October 1988) and lightest in February 1988. order of 20-30% (Barrick et al., 1980; Requejo and
These data appear to reflect a seasonal change in the Quinn, 1983).
balance of organic materials from various sources.
Extractable Lipid Constituents
Horizontal variability in surface sediments Total lipid
Horizontal variability of % organic carbon, total
The amount of total lipid extract was determined
nitrogen, 6 Z3Cocand lipid constituents are presented gravimetrically by weighing the extract after evapo-
in Table 2. The coefficients of variation (C.V.) for
rating excess solvent under a gentle stream of N~.
These results are presented in Table 1 for each of the
Stable i s o t o p e s of s e d i m e n t a r y o r g a n i c c a r b o n
sediment samples. Lipid extracts ranged from 1.8 to
- 1 7 --
5.4mg g-l dry sediment (+20%, as determined in
spatial variability experiment). Lipid normalized to
the organic carbon content of the sediments indicated
?
~o
-18
I t t t tttt t that the lipid content was generally higher during late
winter-early spring months (February and March)
-19
t when new particulate matter was delivered to the
oo
-20
t study site (Table I).

t Fatty acids
The abundance of fatty acids present in the surface
-21 Itll I I I Iit11111
S ND FMA J J A S O D sediments exhibited temporal differences (Table 3).
'87- - - %. . . . '88 . . . . . . . . Concentrations of solvent extractable fatty acids
(C12-C32) ranged from 111-416/ag g-i dry sediment
Fig. 2. Stable carbon isotope ratios (6~3Coc) for total in the surface sediment interval. Total fatty acids
organic carbon in surficial sediments collected monthly from were higher in June-August (~ = 352 +40, n = 5)
Cape Lookout Bight, NC. Months when cores were not
collected have been omitted from the x-axis in this and than at other times of the year (~ = 164 _ 48; n = 7).
subsequent figures. Error bars are as determined during the When normalized to the organic carbon content of
spatial variability study. the surficial sediments, fatty acids were enriched
568 ELIZABETH A . CANUEL and CHRISTOPHER S. MARTENS

Table 2. Results from spatial variability study

Hydrocarbons Sterols Fatty acids

% Org C % Tot N 6~3Coc #g/g dry sediment

2.42 0.30 -18.74 2.44 17.85 284.19
2.59 0.31 --18.54 1.94 12.28 175.05
2.62 0.32 -18.89 2.09 13.05 192.01
2.64 0.32 -18.59 1.63 9.43 213.50
2.48 0.30 - 19.13
2.74 0.34 - 18.56
2.65 0.30 -- 18.80
2.82 0.34 - 18.57
2.73 0.36 - 19.30
2.47 0.31 - 18.97
2.61 0.32 -18.81 2.03 13.15 216.19
s.d. 0.13 0.02 0.26 0.34 3.50 47.99

relative to organic carbon during summer months
[Fig. 3(a)]. Values for FAMEs/OC ranged from
3.3-12.2~g mg -l OC ($ = 7.1_ 2.9, n = 1 2 ) with
summer values (June and August 1987 and
June-August 1988) always above the mean (+20%).
In general the fatty acid distributions were domi-
nated by the 14:0, 16:0, 18:0 and 16:1co7 com-
ponents. However, during summer months 13:0,
15:0 and 17:0 n-, iso- and anteiso-branched car-
boxylic acids increased in abundance. Their concen-
trations ranged between 14.2 and 95.8/~g g-I dry
sediment (see Ct3 , C15 and Ct7 BactFAMEs in Table
3), comprising up to 30% of the total fatty acids
present in the surface sediments during summer
months. When normalized to the organic carbon
content of the surficial sediments, bacterial com-
ponents were enriched relative to bulk organic carbon
during summer months, as well [Fig. 4(c)].
Polyunsaturated C20 and C22 fatty acids (PUFAs)
were most abundant in March, April and June (1987
and 1988). The abundance of these components
ranged between 6.2 and 31.2/tg g-l dry sediment
(Table 3). Concentrations of these constituents were
highest during June 1988 although when expressed as
a percentage of the total fatty acids, PUFAs were
relatively more abundant during February at which
time they accounted for 16% of the total fatty acid
distribution. During spring months, PUFAs were
also enriched relative to the organic carbon content
of the surface sediments [Fig. 4(a)]. Long-chain satu-
rated fatty acids (>C22) accounted for a small per-
centage ( < 5%) of the total fatty acids (Table 3).
Sterols
Sterol concentrations varied by a factor of three
over the study period (15.5-49.8/~g g-i dry sediment;
Table 3). Sterol/OC ratios ranged from 0.53 to
1.46/zg mg -l OC. The highest sterol concentrations,
as well as the highest sterol/OC ratios occurred in the
June 1988, August, March and February samples
[Table 3; Fig. 3(b)]. No distinct temporal trends were
evident in the concentration of total extractable
sterols present in the surface sediments, however,
individual sterol components did show seasonal
differences in their absolute abundance.
Generally, either cholest-5-en-3fl-ol (S-5) or 24-
ethylcholest-5-en-3fl-ol (S-19) was the most abundant
sterol. Concentrations of cholest-5-en-3fl-ol were
highest in June and August 1988 (10.5 and 6.8/zg g-~
dry sediment, respectively) and varied by a factor of
four over the entire study period (2.5-10.5/~g g-~ dry
sediment). The concentrations of both 24-ethyl-
cholest-5-en-3fl-ol and 24-methylcholesta-5,22E-
dien-3fl-ol were generally higher in samples where
sediment accumulation was greatest (i.e. February,
March and June 1988; Table 3).
Hydrocarbons
Concentrations of hydrocarbons ranged between
5.9 and 26.8 #g g-l dry sediment with maxima occur-
ring in February and June 1988 (Table 3). Hydrocar-
bons normalized to organic carbon ranged between
0.3 and 0.78/zg mg -l OC with the February and June
1988 samples again having the highest values [Fig.
3(c)]. Hydrocarbon distributions contained n-alkanes
in the Cl~C3s range. The odd to even relationship
within a homologous series is expressed by the carbon
preference index (CPI). Values calculated according
to the method of Bray and Evans (1961) ranged
between 1.1 and 3.3 over the C23--C35 range, but
showed no seasonality in their distribution (Table 3).
Branched C25 isoprenoid hydrocarbons were abun-
dant in several of the samples (Table 3). The assem-
blage was primarily comprised of branched C25 dienes
and trienes with branched tetraenes present as minor
components. Cyclic components were not identified
in any of the samples. Identifications were based on
comparison of GC-MS data with published spectra
(Barrick et aL, 1980; Requejo and Quinn, 1983;
Requejo et al., 1984). The relative abundance of
individual components within this group varied be-
tween samples suggesting multiple sources.
DISCUSSION
Seasonal Variations in Organic Matter Sources
Potential sources of organic matter in sediment
accumulating at our study site include highly
productive marsh and seagrass environments,
 Table 3. Summary of source-specific constituents. Note: Underlined values are greater than the annual mean +_ 20%
28 Jun 87 13 Aug 87 21 Nov 87 10 Dec 87 27 Feb 88 29 Mar 88 30 Apr 88 19 Jun 88 20 Ju188 17 Aug 88 1 Sept 88 2 Oct 88 .f ( + 2 0 % )
Fatty acids (jag g-t dry sediment)
C20 & Cz2 PUFAs 23.._!1 16.7 10.1 14.3 18.6 22.3 20.._.55 31.._.22 16.9 11.0 12.1 6.2 16.9 (3.4)
Ct3,Cts,Ci7 BactFAs 76.5 90.1 20.8 22.6 14.2 20.3 24.5 85.2 95.8 74.8 65.8 44.0 52.9 (10.6)
C23-C32 sat'd FAs 5.--~ 4.'-~ 0.06 0.3 2.1 1.0 2.0 3.-"~6 5.'~5 0."--9 0.---~ 0.7 2.2 (0.4)
Y-Fatty acids 327.4 364.6 111.3 128.7 114.1 173.2 178.1 416.2 328.2 321.4 238.2 203.6 242.1 (48.4) ~,,
Sterols (lag g-i dry sediment)*
S-3 1.4 1.8 1.8 1.2 2....66 2.1 1.1 4.7 1.4 2._5 1.8 0.9 1.9 (0.4) 2.
S-5 3.9 5.7 5.4 4.0 5.1 5.4 2.5 10.__._55 3.7 6._.88 4.9 3.2 5.1 (1.0) ~.
S-6/S-5 0.34 0.35 0.24 0.26 0.24 0.20 0.26 0.23 0.31 0.33 0.29 0.28 0.28 (0.06) o
S-8 2.5 .'-5"-
3 3.6 2.9 7..__0 7._.99 1.9 5.3 2.5 4.2 3.3 1.8 3.9 (0.8)
S-I 2 + S-16 2.2 3.8 2.0 1.9 6.__[ 5.5 3.1 9.0 3.9 6.7 4.8 2.7 4.3 (0.9) ~"
S-19 3.0 4.4 4.4 2.1 6.._66 7.1 2.9 1._..00
l 3.1 5.4 3.8 2.4 4.7(0.9)
]:Sterols 25.2 35.6 29.5 21.3 40._..88 39.__.~5 15.5 49.____88 21.2 40.___22 28.9 16.5 30.3 (6.1)
Hydrocarbons Olg g i dry sediment ) ~_
CI7 O.I 0.4 0.7 O.1 0.3 0.7 0.2 0...55 0.4 .._.22
1 0.4 (0.08)
C19 0.1 0.1 0.1 0.2 0.1 0...~4 0.2 0.05 0.1 0.1 ._..22
1 0.2 (0.04)
C21 0.2 0.4 0.3 .0.9 0.4 . 0.5 . 0.9 O.
. 1 0.3 0.4 0.9 0.4 (0.08) *~
g:l
C23 O.1 0.4 0.9 0.5 I ._.99 1.3 1.5 2.4 O.1 0.5 1.0 1.0 1.0 (0.2) ch
l~br25: 2 0.2 3.._88 1.5 2._._55 2.._66 0.5 2.7 1.7 1.1 1.5 1.7 0.7 1.7 (0.3) ,.~
• ,br 25: 3 1.4 0.4 0.2 2.4 0.1 1.3 0.3 1.2 0.8 1.7 1.0 0.9 (0.2) t~
Ebr25:4 0.-~ 0.2 1.--6 -- 0.2 0.'-1 0."~ .....0
1 0.25 (0.05)
• C23-C33 9.6 8.5 7.4 7.0 14.2 13._..99 7.2 19.___0 1.7 6.4 8.1 6.1 9.1 (1.8) o
CPI23_3st 3.2 2. I 1.3 1.7 1.3 1.1 1.1 1.3 3.._~3 2.0 1.5 2.8 2.0 (0.4)
ZHydrocarbons 10.0 17.1 11.7 12.0 26.._.88 15.2 12.9 25..__99 5.9 11.8 14.5 13.0 14.7 (2.9) o
Lignin ~ o
C/V 0.8 0.9 I. 1 1.2 1.3 1.1 0.9 0.9 1.__66 1.1 1.1 (0.2)
[Ad/Al]v __0.9 0.5 0.3 0.3 0.5 0.2 0.4 0.6 0.6 0.4 0.5 (0. I) ,~.
Stable isotopes
613Coc -18.7 -19.1 -20.3 -19.6 -18.7 -18.4 -18.4 -18.1 -17.8 -18.7 -18.7(0.3)
*Sterol assignments: S-3 cholesta-5,22E-dien-3fl-ol; S-5 cholest-5-en-3fl-ol; S-6 5a(H)-cholestan-3fl-ol; S-8 24-methylcholesta-5,22E-dien-3fl-ol; S-12 24-methylcholest-5-en-3//-ol; S-16 24-ethylcholesta-5,22E-
dien-3~-ol; S-19 24-ethylcholesta-5-en-3/Lol.
2[]~(odd nC23 to nC33)]
tCP123_35= (variation of calculation by Bray and Evans, 1961).
~(even nC22 to nC34 ) q-5Z(even nC24 to nC36)
~:Lignin parameters as defined in Hedges and Mann, 1979: C/V = cinnamyl/vanillyl phenols; [Ad/AI]v = acid/aldehyde ratio of vanillyl phenols.

~D
570 ELIZABETHA. CANUELand CHRISTOPHER S. MARTENS

Lipid Compound Classes Normalized of organic matter contributing to sedimentary en-

to Organic Carbon vironments. This is due to the equivocal nature of
some biomarker compounds, difficulties in correcting
o
for the effects of diagenetic processes and limited
~oo 10 information regarding the lipid composition of living
organisms. In the data presented below, source as-
U. ::L signments are based on agreement between multiple
P4
indicators covering a range of geochemical stabilities.
In order to establish objective criteria regarding the
J A ND FMA JJASO relative importance of these sources of organic matter
over time, we have required that the concentration of
3- an established biomarker(s) at any point in our study
0 lie outside its mean concentration over the entire
study period by at least 20%. This threshold value
60 (20%) is based upon the results determined during
our spatial variability study and was used in an
1 attempt to separate the effects of temporal variability
U)
from spatial variability.
o
J A ND FMA d JASO
1.0 Source-Specific Fatty Acids Normalized
1.0 to O r g a n i c C a r b o n
0
~o
o
o
o o
,o
0.8 1 i I
0.5 "< c,
E 0,6
0
3: =L g~ 0.4
~4
0.2
0.0
J A ND FMA J J ASO 0.0
'87 ........... I .......... '88 ...... J A N D FMA J JASO

Fig. 3. Lipid compound classes normalized to organic

carbon. The abundance of each compound class was nor- 0.4
°'s I B
O
malized to the organic carbon content of the surface sedi- OO
0.3
~ o
ments. These data reflect similar trends to those seen when tL
constituents were normalized to g dry sediment (see 0.2
Table 3).

0.0
J A ND FMA JJASO
phytoplankton productivity in the offshore waters
and the back-barrier island sounds, eroding shoreline
peats and terrestrial inputs delivered via the numer-
ous river-estuary systems associated with the North O
2
Carolina coast. Previous work, which concentrated
on the organic matter composition of the upper meter
1
of sediment (Haddad, 1989; Haddad and Martens, m =L

1987), identified these sources as derived from a

mixture of marine algae, vascular plants and bacteria.
However, there should be significant temporal vari-
ations in inputs from these sources because of vari-
ations in rates of primary productivity and plant
senescence, as well as physical transport processes
associated with prevailing wind directions and storm
events. The data presented here were used to eluci-
date seasonal differences in the relative contributions
of various sources of organic matter.
Although biomarker compounds provide better
source information than bulk parameters (elemental
and isotopic, for example), the biomarker approach
is also limited in its ability to discern between sources
0
J A ND FMA JJASO
'87 ........... I .......... '88 ......
Fig. 4. Source-specific fatty acids normalized to organic
carbon. Algal (A), terrestrial (B) and bacterial (C) fatty
acids are normalized to the organic carbon content of the
surficial sediments. Algal components (polyunsaturated C20
and C22 carboxylic acids) are enriched during June and
August 1987 and March, April and June 1988. Terrestrial
components (long-chain saturated fatty acids, i.e. >C22) are
highest during summer months (June and August 1987 and
June and July 1988). Fatty acids derived from bacteria
(normal and iso- and anteiso-branched C~3, C~5 and C~7
acids) are enriched during summer months relative to other
times of the year.
 Seasonal variations in the sources and alteration of organic matter 571

Table 4. Summary of source indicatorsfrom literature*

Source ~ ~3Coc Fatty acids Sterols Hydrocarbons
Algae - 1 8 to - 2 2 polyunsaturated Ci6,Ci8,C20,C22 S-3, S-8 (S-10) CI5,C17,CI9, 21:6, brC25
Terr vascular plants -28 saturated C22~C32 S-12, S-16, S-19 C23-C35, odd predominance
Bacteria iso-, anteiso, normal C~3,C~5,C~7
Scagrasses - 1 0 to - 1 4 16:0, 18:1, 18:2 S-19 CI7,CI9,C21
*References given in text.

Algae algae (Nichols et al., 1990; Volkman, 1986). Like the

A summary of constituents indicative of algal
sources (phytoplankton and macroalgae) is presented
in Tables 4 and 5. High concentrations of C20 and C22
polyunsaturated fatty acids (PUFAs) were found in
surficial sediments collected during March and April
1988 and in June 1987 and 1988 (Table 3). Because
of the reactivity of these polyenoic constituents, their
presence has been used as an indicator of viable algal
cells (Shaw and Johns, 1985). While there is no way
to determine whether this is the case here, the in-
creased abundance of the P U F A s does indicate an
input of relatively "fresh" algal organic matter to the
surface sediments during spring. Algal inputs may
also be important during other times of the year (e.g.
summer). However, due to degradation, either in the
water column or at the sediment-water interface,
these inputs are not recorded in the sediments.
During February, March and June 1988 other algal
indicators, the diatom sterols, cholesta-5,22E-dien-
3fl-ol (S-3) and 24-methylcholesta-5,22E-dien-3fl-ol
(S-8) (Gillan et al., 1981; Volkman et al., 1980; Orcutt
and Patterson, 1975; Tornabene et al., 1974) are
abundant, as well. Like the polyunsaturated fatty
acids, concentrations of these sterol components were
highest during spring months (March and June 1988;
Table 3) supporting the idea that they too are derived
from a spring influx of algal organic matter. These
components were also abundant in the February
sample.
The C29 sterol 24-ethylcholest-5-en-3fl-ol (S-19) is
generally thought to be synthesized by higher plants
(Bae and Mercer, 1970; Wannigama et al., 1981).
However, this component has also been reported in
diatom sterols described in the previous paragraphs,
concentrations of these sterols are highest during
February, March and June 1988 suggesting that this
component (S-19) may be mainly derived from algae
rather than vascular plant-derived organic matter.
The concentration of S-19 was significantly correlated
with the concentrations of both the C20 and C22
PUFAs, as well as S-8 (P = 0.02; P = 0.005, respect-
ively) indicating that S-19 may be derived from algae
in this system. Further support for an algal origin for
the C29 sterols in the surface sediments is shown by
the fact that 24-ethylcholest-5-en-3fl-ol was the most
abundant steroi in particulate matter filtered from
seawater collected from the upper meter of CLB
(E.A. Canuel, unpublished data). The abundance of
lignin oxidation products in the surficial sediments
was also low in February, March and June (Canuel,
1992) further suggesting an algal origin for this
component (S-19) in the surface sediments.
An increase in the abundance of short-chained
n-alkanes (C15--]-C17--[-C19) during February and
June 1988 (Table 3) provides additional evidence for
the importance of algal-derived sources of organic
matter during the late winter-spring months (Blumer
et al., 1971). Short-chained n-alkanes were most
abundant in October, however, when other algal
indicators were low suggesting an alternative source
for these components at this time. Recent work
indicates that branched isoprenoid alkenes may
be derived from diatoms (Nichols et al., 1988;
J. Volkman; personal communication). Like the algal
indicators described above, branched C25 isoprenoid
alkenes were most abundant in February, April and
August 1987 suggesting they are derived from phyto-

Table 5. Biomarker components of N. Carolina Flora

613Coc Fatty acids* Sterols* Hydrocarbons* Lignint
Macroalgae
Codium sp. -- 12.7 18:1,16:0,16:1,20:5,18:0 S19,S10,S16~ ND
Dicyota sp. -- 18.2 18:1,16:0,16:1,20:5,14:0 S19,$5,S12 ND
G. tikvahiae - 15.9 20:5,16:0,18:0,14:0,18:1 S5,S11,S9 ND
G. verracosa - 14.3 18 :l,16:0,14:1,20:1,14:0 $5,$9,SI0 ND
Hypnea sp. -17.1 16: 2,16:0,16:1,14:0,18:1 $5,S16,S19 ND
Neoghardiella sp. -18.5 16:0,20: 5,20:1,16:l,18:0 SI0,S8,SI l ND
Ulva sp. -22.2 16:0,18 : 1,15:0,18:0,14:1 S19,$5,$8 ND
Seagrasses
H. wrightii -9.3 18:1,16:1,16:0,18:0,14:0 S19,S16,S12 C23,C24,C25 C phenols§
Z. marina - 10.0 18:2,16:0,18:1,16:2,16:1 S19,S16,S12 CI9,C21,C23 C phenols
Marshgrasses
S. alterniflora - 12.6 18:1,16:0,18:0,14:0,20:0 S19,S12,S16 ND C phenols
*Data are given for most abundant constituentsin descendingorder of abundance.
"~Datafrom Haddad and Martens, 1987.
:[:Sterol assignments: S-5 cholest-5-en-3fl-ol;S-8 24-methylcholesta-5,22E-dien-3/Lol;S-9 24-methyl-5a(H)-cholest-22E-en-3fl-ol;S-10
24-methylcholesta-5,24(28)-dien-3fl-ol;S-I 1 24-methyl-5,,(H)-cholest-24(28)-dien-3/~-ol; S-12 24-methylcholest-5-en-3//-ol;S-16 24-ethyl-
cholesta-5,22E-dien-3fl-ol;S-19 24-ethylcholest-5-en-3fl-ol.
§Cinnamyl phenols as defined in Hedgesand Mann, 1979.
572 ELIZABETHA. CANUELand ~ p n E l t
plankton. Lastly, an increase in algal organic matter
is supported by the stable isotope data (Table 3;
Fig. 2). A 6~3Coc shift towards a lighter isotopic
signal is seen in late winter/spring months with the
313Coc signal reaching its lightest values in February
and March ( - 2 0 . 3 and -19.6%o, respectively).
Marine plankton in the southeast have a mean 6 t3Coc
value of - 2 2 + 1.6%o (Haines, 1976).
The relative contribution of this "fresh" algal
material to the organic carbon content of the surface
sediments can be estimated making the following
assumptions and using the concentration of the PU-
FAs in the surficial sediments. Polyunsaturated fatty
acids (PUFAs) make-up approx. 4-19% of the total
fatty acids in diatoms (10%, on average) and fatty
acids account for about 40% of their lipid content
S e d i m e n t a c c u m u l a t i o n / d e l i v e r y of
" 1.5 A organic matter
o
.~ 1.0
ig
~ 0.5
m 0.0
J AS ND FMA
15
B event is excluded from the calculation of a correlation
0 % OC ( A L G )
• % OC (VASC)
1o
o
0
o
0 Terrestrially-derived organic matter has been
0
5 -- 0 o
0
o 0
0 o
• Q
I I I I I_1_1 I*l.l*l
A ND FMA
'87 . . . . -4 . . . . .
Fig. 5. Sediment accumulation and % OC derived from
marine algal vs terrestrial vascular plant sources. The ac-
cumulation of new sediment occurs episodically (A) with
most of the annual accumulation occurring during win-
ter/spring months. Months labelled "0" indicate periods
when cores were collected but there was no significant
accumulation as determined using Be-7 (Canuel et al., 1990).
During September 1988 the Be-7 inventory was less than
expected after correcting the previous month's inventory for
radioactive decay indicating removal (R). Organic carbon
derived from algal sources (©) appears to increase during
periods of sediment accumulation (B). This does not appear
to be the case for organic carbon derived from terrestrial
vascular plant sources (0). Calculation of these values (%
OC by source) is explained in the text.
S. MARTENS
(Volkman et al., 1989). If we assume that diatom
biomass is 10% lipid and that the carbon content of
this biomass is 50% on a dry weight basis, the
concentration of PUFAs can be used to quantify the
relative contribution of this "reactive" algal-derived
component to the sedimentary organic carbon [Fig.
5(b)]. "Fresh" aigal-derived organic matter makes-up
2-10% of the organic carbon content of the surface
sediments with values significantly ( > 20%) above the
mean (~ = 5.0, n = 12) occurring in the March, April
and June 1988 samples. Using a more stable indicator
of algal inputs, 24-methylcholesta-5,22E-dien-3fl-ol
to quantify the relative abundance of organic carbon
derived from algal sources indicates an increase in
algal organic matter during February, March and
June 1988. In both cases sediments collected in March
and June 1988 appear to be enriched in this "fresh"
algal source. Months in which either of the algal
indicators were enriched were also characterized by
sediment accumulation (Table 1; Fig. 5) suggesting
that the delivery of algal organic matter may be
associated with horizontal transport processes
(advection from the back-barrier island sounds, for
example). However, it is important to keep in mind
that the delivery of algal organic matter is dependent
on both the abundance of algal organic matter avail-
able for transport, as well as the physical conditions
appropriate for the delivery of sediment. As a result,
other periods of time characterized by sediment ac-
cumulation (October 1988 for example) may not be
characterized by increases in algal organic matter
J JASO
(Fig. 5) if algal-derived organic matter is not available
for delivery. In fact, when the October accumulation
coefficient for %OC from algae vs sediment accumu-
lation, we find there is a significant relationship be-
tween these parameters (r = 0.83; n = 11; p < 0.01).
Terrestrial vascular plants
shown to be a minor source of organic matter
o accumulating in CLB sediments (Haddad, 1989;
Haddad and Martens, 1987). Surficial sediment
o
samples collected during this study contained low
I°l I.I I.I
JJASO
concentrations of lignin oxidation products (Canuel,
1992), long-chained ( > C22) saturated fatty acids and
88 . . . . odd-numbered n-alkanes in agreement with these
earlier findings. Lignin is exclusively associated with
vascular plant tissues where it acts as a structural
component. Long-chain n-alkanes and fatty acids
are generally associated with waxy leaf coatings
(Wannigama et aL, 1981) although they may be
present in diatoms at low concentrations (Volkman
et al., 1980). Long-chain fatty acids (>~nC22) are low
in concentration ( < 6 #g g-t dry sediment, Table 3)
and exhibit temporal variations in their abundance in
the surface sediments (Table 3). These components
accounted for less than 5% of the total fatty acid
distribution throughout the 18 month study period.
Relative to organic carbon, these components were
 Seasonal variations in the sources and alteration of organic matter
most abundant during summer months [June and
August 1987 and June and July 1988; Fig. 4(b)]
suggesting that their preservation was enhanced rela-
tive to more reactive constituents removed during
microbiaily-mediated degradation processes.
The distribution of the long-chain fatty acids
(C23-C32) with a strong even/odd predominance can
be used to estimate the terrestrial component of the
organic carbon in CLB surface sediments as was done
in the previous section for the algal component.
Again, this calculation requires some assumptions
regarding the composition of terrestrial vascular
plants. We assume, for example, that long-chain acids
make-up about 4% of the extractable lipids in higher
plants (Shaw and Johns, 1985) and that the biomass
of higher plants is ~0% carbon with 10% of this
carbon represented by lipids. Estimating the portion
of the organic carbon derived from terrestrial
sources in this way indicates that a relatively low
percentage of the organic carbon is represented by
terrestrial vascular plants ( < 2 % total OC). Unlike
organic carbon derived from algal sources, the
abundance of terrestrial organic carbon does not
appear to be related to sediment accumulation
(Fig. 5).
Seagrasses
Using a combination of lignin oxidation product
techniques and stable carbon isotopes, Haddad and
Martens (1987) estimated that 23 _ 17% of the or-
ganic carbon supplied to this site was derived from
non-woody angiosperms (probably S. alterniflora and
the seagrasses Z. marina and H. wrightii). The prox-
imity of marshes and seagrass beds to the study site
(Fig. 1) and high rates of export from areas near CLB
(Bach et al., 1986) suggests that these plants may be
important sources of organic matter.
A study of lipid constituents present in various
coastal macrophytes collected from the vicinity of
Cape Lookout yielded specific biomarkers for the
seagrasses (Table 5). In general, fatty acids and
hydrocarbons derived from the seagrasses were simi-
lar to those expected for algae rather than for terres-
trial vascular plants. Sterol constituents were
dominated by 24-ethylcholest-5-en-3fl-ol with the
stereochemistry at C-24 unknown. The hydrocarbon
distribution of Zostera marina contained relatively
large concentrations of short-chained n-alkanes
(C17-C23), more usually attributed to phytoplankton
(Table 5). High concentrations of these low molecular
weight n-alkanes have been measured in Zostera
marina during its flourishing stage and these hydro-
carbons have also been found to be important con-
stituents in Australian seagrasses (Khandekar and
Johns, 1990; Nichols et al., 1982 and references cited
therein; Volkman et al., 1980).
The C~7-C23 n-alkanes were distinctly more abun-
dant in surface sediments collected during October
1988 than for any other time during the study (Table
3). However, in contrast to other times of the year
573
when these short-chained n-alkanes were enriched
(i.e. February and June), other algal indicators (i.e.
the C20 and C22 PUFAs and algal sterols, S-8 for
example) were not enriched in October. This suggests
an alternative source for these n-alkanes in the
October sample.
During both years of the study, the 6 ~3Coc values
were heaviest during late summer-early fall (Septem-
ber 1987 and October 1988; Fig. 2; Table 3). Sea-
grasses fractionate carbon similarly to C-4 plants,
resulting in a heavier isotopic signature for the sea-
grasses ( - 10.0 to 14.0%o;Thayer et al., 1978) than for
most of the other sources of organic matter available
to the study site. Thus, an increase in the input of
seagrass-derived organic matter should shift the iso-
topic signature of the total organic carbon towards a
heavier signal as is seen in the October sample
(-17.8%o). Further support for a seagrass input
during autumn is seen by the increase in cinnamyl
phenols, the most abundant lignin-derived phenols in
these plants, during September/October 1988 (see
C/V or cinnamyl/,anillyl ratios in Table 3). Lastly,
although the seagrasses die-off during warm summer
months (Thayer et al., 1984), their delivery into CLB
is dependent on physical forces. Autumn months are
characterized by an increase in north/northeast winds
relative to summer months which are dominated by
winds from the south/southwest. The frequency of
north/northeast storm activity has been proposed as
a mechanism for delivering particulate matter to CLB
(Canuel et al., 1990). Sediment accumulation was
measured during September 1987 and October 1988
and removal (which is not inconsistent with storm
activity) in September 1988 [Table 1; Fig. 5(a)]. Thus,
several source indicators: the heavy isotopic signature
of the surficial sediment organic carbon pool, the
increase in cinnamyl phenols and the increased abun-
dance of the C~7-C23 n-alkanes, as well as the appro-
priate conditions for delivery to this site, suggest that
the seagrasses may be an important source of organic
matter during autumn months (e.g. September/
October). Other techniques (compound specific
isotope ratio mass spectrometry) are presently
being used to assess the potential importance of sea-
grasses as sources of organic matter to the sediments
of CLB.
Bacteria
Bacterially-derived fatty acids exhibited strong sea-
sonality with their maxima consistently occurring
during summer months (June-September; see Table
3). These high concentrations of bacterial fatty acids
reflect the high rates of microbial processes such as
sulfate reduction measured by others at this site
during summer months [Fig. 6(a), Crill and Martens,
1983, 1987]. The abundance of the odd-numbered
normal and branched bacterially-derived carboxylic
acids ranged from 6.0 to 42.3 and 4.6 to 22.1/~g g-~
dry weight sediment, respectively. During summer
574
Sulfate reduction/ %bacterial Org. carbon
40- A
30
%.. ~ 0 ° 0 •
20 - o
0 0
_ • ~ . ~
[.., tO o o Temp
• SRR
I I Ill II)l I I Illll
J A SONDJ FMAMJ JASO
15 m
B sulfate reduction (Fig. 6; Crill and Martens, 1987).
U 10 m dependent rates of sulfate reduction increase (Fig. 6).
2
© 5 --
) I Ill II)t I I IIIlll
A ND F M A J J AS O
'87 . . . . -I . . . . .
Fig. 6. Sulfate reduction rates and % organic carbon from
bacteria. Sulfate reduction rates (SSR; data from Crill and
Martens, 1987) and temperature (data from Chanton, 1985)
exhibit dramatic seasonal cyclicity in the sediments of Cape
Lookout Bight, NC (A). The fraction of the organic carbon
that can be attributable to bacterial sources increases four-
fold during summer months in response to increases
in temperature-dependent rates of bacterial sulfate reduc-
tion (B).
months, a single peak (consisting of the co-eluting
anteiso-17:0 and 17:1 acids)accounted for 40% of
these bacterial fatty acids. It is thought that these
components are derived from the abundant sulfate
reducers present in the surface sediments (Taylor and
Parkes, 1983; Parkes and Taylor, 1983; Boon et al.,
1977; Edlund et al., 1985).
Bacterial components such as the odd-numbered
normal and branched (iso- and anteiso-) fatty acids
(see Cl3, Cls, Cl7 BactFAs: Table 3) can be used to
estimate the relative importance of bacterial organic
carbon. These components make-up approx. 35% of
the fatty acid content of sulfate-reducing bacteria
(Parkes and Taylor, 1983; Taylor and Parkes, 1983).
If we assume bacterial biomass is 50% carbon
with 20% of the carbon present as lipids we can
estimate the bacterial contribution to the sedimentary
organic carbon pool [Fig. 6(b)]. From these cal-
culations, we see that bacterial sources make-
up approx. 10% of the organic carbon during sum-
mer when rates of sulfate reduction are highest
[Fig. 6(b)].
ELIZABETH A. CANUEL and CHRISTOPHER S. MARTENS
Seasonal Importance of Microbially-Mediated
Processes
The distribution of bacterial fatty acids suggests that
~ 0 O° • microbial biomass may be a significant component of
the organic carbon content of these sediments during
~ summer months. Fatty acids specific to bacteria
increase during summer months when there was little
_
or no new accumulation of sediments. For example,
::1 x - ~ there was no sediment accumulation during Summer
1987 and accumulation during only May/June in the
Summer 1988 [Canuel et al., 1990; Fig. 5(a)]. The
concentration of bacterially-derived components in
the surficiai sediments was also strongly correlated
(r2=0.86) with seasonality in integrated rates of
Organic carbon associated with bacterial sources
increases during summer months when temperature-
These data suggest that changes in the fatty acid
distribution reflect remineralization processes result-
ing from microbial activity within the CLB sedi-
ments.
Bacteria not only contribute their own metabolic
constituents but alter the signature of organic matter
via microbially-mediated diagenetic transformation
reactions. One such reaction is the reduction of
cholest-5-en-3fl-oi to 5~t(H)-cholestan-3fl-ol (Gaskell
88 . . . . and Eglinton, 1975; Nishimura and Koyama, 1977).
The ratio of 5~t(H)-cholestan-3fl-ol to cholest-5-en-
3fl-ol in the surficial sediments of CLB was higher
during summer (2 -- 0.32 + 0.04, n --- 6; see Table 3,
Fig. 7) than at other times of the year (~ = 0.25 ___
0.03, n = 6). Although direct inputs of 5ct(H)-stanols
from marine organisms such as diatoms are possible
(Volkman et al., 1990), the increase in the 5ct(H)-
cholestan-3fl-ol to cholest-5-en-3fl-ol ratio correlates
with high rates of sulfate reduction and the pro-
duction of bacterial fatty acids rather than PUFAs or
other algal biomarkers suggesting it is the result of
microbial processes. Dramatic differences other in-
vestigators have measured across redox boundaries
such as those occurring at oxic-anoxic interfaces
(Wakeham, 1989) are not evident here, however, it
should be noted that we are evaluating this process
over relatively short timescales (i.e. on the order of
months to seasons). A t-test of the ratio of 5ct(H)-
cholestan-3fl-ol to cholest-5-en-3fl-ol indicates that
values for summer months are significantly different
than those seen during other seasons (P = 0.01).
Microbial activity appears to influence the organic
carbon content of the uppermost surface sediments
over relatively short periods of time (i.e. during
summer, see Fig. 6). This is shown by high concen-
trations of bacterial fatty acids during summer
months when ailochthonous inputs of particulate
matter are negligible, rates of sulfate reduction are
high and there is good evidence for microbially-medi-
ated transformation reactions. It should be noted that
concentrations of total fatty acids track the temporal
 Seasonal variations in the sources and alteration of organic matter
5a(H)-Cholostan-30-ol/Cholost-5-on-38-ol
A from the uppermost 0.5 cm during this study. In the
o 0.35"
o 17:1). This is not surprising considering that the core
•. 0.25'
0.15
JASONDJ FMAMJ JASO
'87 .......... I ........ '88 ........
Summer vs. Non-Summer Months
0.4
o
=(B 0.3
w
"6
0,2
=o
0.1
(n
0.o
Summer non-Summer
Fig. 7. The ratio of 5ct(H)-cholestan-3fl-ol to cholest-5-en-
3fl-ol in the surficial sediments (0-0.5 cm) of cores collected
during this study are presented for each month when
samples were analyzed (A). Values are higher during sum-
mer than in non-summer months. This is further illustrated
in (B) where the mean of the ratio of these components is
presented for summer (~ = 0.32) vs non-summer (£ = 0.25)
sample periods.
variations seen for the bacterial fatty acids suggesting
that other fatty acids are probably derived from
bacterial sources [see Fig. 3(a) and 4(c)]. The data
suggest that bacteria are capable of altering the
source signature of organic components present in
the surficial sediments of CLB and indicate that the
depositional record can be biased by microbial ac-
tivity even over geochemically short timescales (i.e.
months). It appears that in situ diagenetic processes
may be more important than temporal variations in
delivery in determining the overall composition of
sedimentary organic matter at this site.
A Comparison with the Longer Depositional Record
Previous work conducted in CLB sediments exam-
ined source inputs over a 10yr time period using
hydrocarbon, fatty acid and lignin oxidation product
distributions (Haddad, 1989). The steady-state
nature of the study site on an annual timescale allows
us to compare data collected during our relatively
short study (18 months) with data collected from the
upper 5 cm of a core spanning several decades.
OG20/;~-E
575
Ratio Data collected from the surface interval (0-5 cm) of
Haddad's core were compared with data collected
0-5 cm section, 28% of the fatty acids are the bac-
terial-derived components (i.e. iso- and anteiso-
branched acids, C13-C19 odd-numbered acids and
was collected during July, when our data showed that
30% of the fatty acids were these bacterial com-
ponents. However, higher inputs of polyunsaturated
fatty acids (PUFAs) during winter-spring months are
averaged out when the sediments are examined over
a longer timescale (i.e. with less depth resolution). In
the two surfacemost sediment intervals of Haddad's
core (0-5 and 5-10cm), PUFAs comprised only
3-5% of the total fatty acids indicating that substan-
tial degradation of these compounds may have
occurred.
Haddad (1989) concluded that the major sources of
the large unreactive fraction ( ~ 7 9 % ) of organic
matter accumulating in the sediments of CLB were
reworked marine algae and bacteria, as well as a
small input of terrestrially-derived organic matter.
From the distribution of lipid components, he pro-
posed that the remaining reactive portion ( ~ 2 1 % )
was derived from marine algae/bacterial sources. The
results presented here suggest that most of the reac-
tive fraction of the organic matter accumulating in
the sediments of Cape Lookout Bight, NC is algal in
origin and that this portion is delivered primarily
during spring. Bacterial organic matter produced
during in situ processes may also make-up a portion
of the "reactive" carbon at this site. An additional
source of labile organic matter may be contributed by
seagrasses during late summer/fall. It appears that it
is the availability of "reactive" carbon (algae, bac-
teria and/or seagrasses) which "fuels" the high rates
of microbial activity seen during summer months.
This results in the transfer of algal/seagrass carbon
into microbial biomass and it is a combination of
these altered algal and bacterial signals which is
subsequently preserved in the sedimentary record.
CONCLUSIONS
Seasonal variations in the sources of organic mat-
ter accumulating in coastal sediments may provide an
important control on biogeochemical processes oc-
curring at or near the sediment-water interface. In
the surficial sediments of CLB, lipid constituents
indicative of algal organic matter were more abun-
dant during spring months. These months were also
characterized by higher sediment accumulation. This
spring influx of labile organic matter appears to
"fuel" the high rates of microbial activity seen during
subsequent summer months.
Summer months were characterized by high con-
centrations of bacterially-derived lipids and evidence
for microbially-mediated transformation processes.
Bacterial fatty acids (normal and branched Cl3, C~5
576 ELIZABETHA. CANUELand CHRISTOPHERS. MARTENS
and C17 ) were threefold more abundant during sum-
mer months. High concentrations of these biomarker
compounds and an increase in the ratio of 5at(H)-
cholestan-3fl-ol to cholest-5-en-3/~-ol were highly
correlated with integrated rates of sulfate reduction.
Organic carbon derived from bacteria was also most
abundant during months when temperature-depen-
dent rates of sulfate reduction were highest.
Microbially-mediated degradation processes ap-
pear to have a significant effect on the composition of
sedimentary organic matter in coastal sediments. The
data presented in this paper demonstrate that lipid
constituents can be diageneticaily altered over short
geochemical timescales. As a result, the source signa-
ture of labile constituents may be lost and/or altered
and the sedimentary record significantly biased at
timescales of a year. Our data indicate that micro-
bially-mediated processes occurring in situ may con-
trol the composition of preserved organic matter to
a greater extent than do temporal changes in the
delivery of materials derived from allochthonous
sources.
Acknowledgements--We would like to extend our thanks to
Dan Albert, Marc Alperin, Jeff Chanton, Emmett Duffy and
Frank Wilson for their assistance with field work and
monthly core collections using SCUBA. Larry Benninger,
Mary Jo Baedecker and Stuart Wakeham provided
thorough reviews of earlier versions of this manuscript, as
well as helpful discussions during the development of this
project. Comments from M. I. Venkatesan and an anony-
mous reviewer improved the manuscript. Special thanks are
extended to Cheryl Kelley and Neal Blair for their assistance
in obtaining the ~13Coc data. Financial assistance was
provided by NSF grants OCE87-16528 and OCE90-17979
(CSM), a Sigma Xi grant-in-aid of research (EAC) and a
Limited Service Traineeship from the University of N.
Carolina (EAC).
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