Research in Microbiology 156 (2005) 457–464

www.elsevier.com/locate/resmic

Mini-review

Anaerobic ammonium oxidation (anammox) in the marine environment

Tage Dalsgaard a,∗ , Bo Thamdrup b , Donald E. Canfield b
a National Environmental Research Institute, Department of Marine Ecology, Vejlsøvej 25, P.O. Box 314, 8600 Silkeborg, Denmark
b Danish Centre for Earth System Science, Institute of Biology, University of Southern Denmark, Campusvej 55, 5230 Odense M, Denmark

Received 27 October 2004; accepted 19 January 2005

Available online 17 March 2005

Abstract
Anammox, anaerobic ammonium oxidation with nitrite, is now recognized as an important process in the marine nitrogen cycle. The
bacteria conducting anammox are highly specialized and appear to belong to the Planctomycetales. The process has now been found in a
range of environments including marine sediments, sea ice and anoxic water columns, and it may be responsible for up to 50% of the global
removal of fixed nitrogen from the oceans.
 2005 Elsevier SAS. All rights reserved.

Keywords: Anaerobic ammonium oxidation; Anammox; Review

1. Introduction process was energetically favorable [2,3], yet another reason

to expect it might occur in nature.
Already decades ago, there were several indications that In the early 1990’s the first direct evidence of the anaer-
NH+ obic oxidation of NH+ 4 came from a waste water treatment
4 could be oxidized anaerobically in nature. For exam-
ple, in 1941 anaerobic NH+ 4 oxidation (now called anammox
facility in Delft, The Netherlands [36]. It proved difficult to
isolate the process in the laboratory, but after a longer period
when coupled to NO− 2 reduction as explained in Section 3)
of trial and error anammox was running in a laboratory-scale
was advanced as a possible source of N2 in the sea [11].
fluidized bed reactor [17]. A few years after the initial dis-
Later observations from anoxic water columns indicated that
covery of anammox, the first direct evidence for anaerobic
NH+ 4 disappeared under anoxic conditions [3,20,21], and the NH+ 4 oxidation in a natural environment, marine sediments,
oxidation of NH+ −
4 with NO3 to form N2 was the explana- was published [33]. Also, the initial suggestions of anaer-
tion most consistent with the data. Similar observations were obic NH+ 4 oxidation in anoxic water columns (e.g., [20])
made in the anoxic sulfide-free portion of the water column have been corroborated by the discovery of anammox in the
in the Black Sea [18]. Here, NH+ 4 transported from below anoxic water column of a coastal bay in Costa Rica, Golfo
disappeared before it met the O2 in the overlying water, and Dulce [6], and in the anoxic water column of the Black Sea
there was a ca. 20 m thick zone free of both O2 , NH+ 4 and [15]. Here, we will review the current knowledge about the
H2 S. Nitrate was present here, and it was suggested that anammox process in the marine environment, also drawing
NH+ −
4 was oxidized with NO3 , preventing the NH4 from
+
from knowledge on anammox in waste water treatment.
meeting the O2 . In marine sediments the oxidation of NH+ 4
with NO− 3 had also been invoked to explain the anoxic dis-
appearance of NH+ 4 [1,16,27]. Moreover, calculations of
G
2. The anammox bacteria
values for the oxidation of NH+ −
4 with NO3 showed that the
2.1. Cell biology

* Corresponding author. Anammox bacteria are extremely difficult to isolate and

E-mail address: tda@dmu.dk (T. Dalsgaard). thus far no pure cultures have been obtained. However,
0923-2508/$ – see front matter  2005 Elsevier SAS. All rights reserved.
doi:10.1016/j.resmic.2005.01.011
458 T. Dalsgaard et al. / Research in Microbiology 156 (2005) 457–464
some highly enriched cultures have been established from
waste water treatment facilities, and from these we under-
stand a great deal of both the biochemistry and the cell
biology of the bacteria. All known anammox bacteria have
a membrane-bound compartment in the cell, known as the
anammoxosome, in which the anammox process is believed
to take place [13,38]. The membrane of the anammoxosome
contains ladderane lipids which form an exceptionally tight
barrier against diffusion and which appear to be unique to
anammox bacteria [8]. The hypothesis is that enzymes in the
membrane catalyze the oxidation of NH+ −
4 with NO2 , with
hydrazine and hydroxylamine as intermediates, and a pro-
ton motive force is created across the membrane, which is
used for ATP production. The very tight membrane structure
limits the diffusion of protons across the membrane enhanc-
ing ATP production. It also prevents loss of the reaction
intermediates, and it confines the very reactive intermediate,
hydrazine, to the anammoxosome and thus prevents it from
doing damage to the rest of the cell [38].
2.2. Identity
The first discovered anammox bacterium belongs to the
Planctomycetales [28] and, while not in pure culture, was
named Candidatus Brocadia anammoxidans. Subsequently,
waste water treatment plants have yielded other anammox
bacteria, and to date three genera have been described:
Brocadia, Kuenenia and Scalindua [26]. Within the genus
Scalindua, two marine molecular isolates have been identi-
fied. One of these, found in the anoxic water column of the
Black Sea [15], has been given a tentative species designa-
tion S. sorokinii, and another has been found in the sediment
of Randers Fjord [22]. In both these locations the anammox
process was documented by the use of 15 N compound la-
belling experiments (see Section 3.1). In the Black Sea the
occurrence of anammox bacteria was further substantiated
by the presence of the unique ladderane lipids. Thus, the
cell structural properties of anammox bacteria appear similar
in waste water treatment plants and in nature. However, the
occurrence of the anammox process and the simultaneous
detection of Scalindua, does not rule out the possibility that
other phylogenetically unrelated organisms may take part in
the anaerobic oxidation of NH+ 4 in nature.
2.3. Growth of anammox bacteria
Nothing is known about the growth rate of marine anam-
mox bacteria, but the related bacteria from waste water treat-
ment facilities grow slowly with doubling times of around
9 days under optimal conditions [29]. The optimal tempera-
ture for the waste water treatment organisms is 37 ◦ C [14],
and one might expect even slower growth for anammox bac-
teria living at much lower temperatures as generally encoun-
tered in the marine environment. However, the temperature
optima for marine anammox populations appear to conform
to the local environment. For example, in the permanently
cold sediment of Young Sound, Greenland, with a temper-
ature of < −1 ◦ C year round, the temperature optimum for
anammox was 12 ◦ C [25], and in the Skagerrak, with annual
temperatures between 4 and 6 ◦ C, the optimum temperature
was 15 ◦ C [7]. We await the isolation or enrichment of ma-
rine anammox bacteria for a better understanding of their
growth characteristics.
3. Characteristics of marine anammox
3.1. Documentation and stoichiometry
The discovery of anammox in marine sediments, and
the subsequent identification of anammox in various ma-
rine environments, was based on experiments using a 15 N
isotope labelling technique [33]. These experiments are
typically performed with anoxic homogenized sediment or
anoxic water, and a preincubation is employed to consume
any O2 , and, in some cases, also the NO− −
3 or NO2 initially
present. Three different amendments are then made in par-
allel: 15 NH+4 (A),
15 NH+ + 14 NO− (B), and 15 NO− (C).
4 3 3
After a suitable incubation time, the N2 gas is extracted and
analyzed on a mass spectrometer for the specific labelling
of 15 N in N2 gas (14 N15 N and 15 N15 N). The production of
15 N labelled N in amendment A would require the oxi-
2
dation of NH+ −
4 with oxidants other than O2 , NO3 or NO2 ,
−
which are not found in the incubation. Such a production has
thus far not been observed (e.g., [32,33]). Any production
of 14 N15 N in amendment B must thus be due to coupling
of the 15 N in NH+ 4 with the
14 N in NO− and that is evi-
3
dence of anaerobic NH4 oxidation. Since NO−
+
3 is readily
reduced to NO− 2 in anoxic marine sediments and waters (see
− −
below) it may be either NO3 or NO2 which is the direct ox-
idant of NH+ 4 . Having identified the anaerobic oxidation of
NH+ 4 with NO − −
3 or NO2 in amendment B, the relative im-
portance of this process for N2 production, as well as that
of denitrification, are estimated with amendment C. Thus,
in amendment C the 15 N labelling of the NO− 3 pool is very
high, and denitrification will mostly produce N2 gas labelled
15 N15 N, combining 2 15 N atoms from the added 15 NO− .
3
Also, in amendment C the oxidation of NH+ −
4 with NO3 or
NO− 14 N15 N by the combination of
2 will mainly produce
14 N from NH+ and 15 N from NO− . There will sometimes,
4 3
however, be some native 14 N-labelled NO− 3 in amendment
C incubations, but by knowing the exact 15 N labelling of
the NO− 3 pool, the contribution of anaerobic ammonium
oxidation and denitrification to N2 production can still be
estimated [33].
The question now is how to identify the oxidant for am-
monium. The reduction of NO− 3 to the oxidation state of N2
requires five electrons, while three electrons are liberated in
oxidizing NH+ 4 to N2 yielding a 5:3 stoichiometry for the
reaction in Eq. (1).
3NO− + +
3 + 5NH4 → 4N2 + 9H2 O + 2H . (1)
 T. Dalsgaard et al. / Research in Microbiology 156 (2005) 457–464 459

With this stoichiometry, 1 out of 4 N2 molecules would be

formed solely from NH+ 15 15
4 which would give 25% N N la-
+
belling in amendment B (if no native NH4 is present). This
labelling percentage, however, has not been observed. Al-
ternatively, if NO− − −
3 were reduced to NO2 by NO3 -reducing
−
bacteria in the environment, and if this NO2 was the oxidant
of NH+ 4 , the oxidation state changes would be +3 and −3
respectively and the stoichiometry would be 1:1 (Eq. (2)).
NO− +
2 + NH4 → N2 + 2H2 O. (2)
This would explain the observed 14 N15 N
production stoi-
chiometries as typically found with amendment B, and the
ability for NO− +
2 to act as oxidant of NH4 was confirmed
− +
in experiments where only NO2 and NH4 were present [7].
The overall stoichiometry of anaerobic NH+4 oxidation in the
marine environment is therefore the same as found for the
anammox process operating during waste water treatment
[35]. This indicates that the anammox process is responsible
for anaerobic ammonia oxidation in the marine environment
[6,7,22,25,33,34].
Fig. 1. Schematic representation of the distribution of selected dissolved
3.2. Production of NO−
2
species in the sediment porewater and the water above the sediment (A)
and indication of the depth intervals in which selected processes occur (B).
Concentrations are not drawn to scale.
Nitrate is generally much more abundant in the marine
environment than NO− 2 , and the anammox process must rely
on other processes to reduce NO− − during the later part of the experiment when NO− 3 is limiting
3 to NO2 . Yet, this may
− (e.g., [7]).
not limit the rate of anammox as NO3 is readily reduced to
In natural sediments the situation may be similar, but
NO− 2 in suboxic marine environments. In the sediments of with NO− −
3 limitation occurring spatially. Thus, NO3 con-
the Skagerrak (located between Denmark and Norway), the
production of NO− − centrations are highest in the upper part of the anoxic NO− 3-
2 by NO3 reduction in anoxic sediment in-
cubations was 4 times faster than NO− containing zone of the sediment (Fig. 1), and NO− -utilizing
2 consumption [7]. In
3
similar incubations with arctic sediment, the NO− organisms here are likely NO− 3 saturated. Only in the lowest
2 produc-
tion rate was also faster than its consumption rate [25], while part of the NO− −
3 zone does NO3 limitation probably oc-
in the Thames estuary, NO− cur. Nitrate reducers become NO− 3 limited at concentrations
2 production balanced its con-
sumption [34]. In Randers Fjord sediment anammox rates below 2–3 times their Km value for NO− 3 uptake. Assum-
derived from 15 NO− 15 −
3 and NO2 additions were similar, in- ing Km values on the order of 1 µM, or even less [5], it
dicating that production of NO2 from NO−
−
3 was not limiting
would seem that NO− −
3 reduction becomes NO3 limited be-
−
anammox [22]. low 2–3 µM NO3 . In intact sediment from Randers Fjord
−

Most published studies of anammox in marine environ- (Denmark) the NO− −
3 + NO2 ( NO3 ) concentration at the
ments have been based on anoxic incubations of homoge- oxic–anoxic interface was around 20 µM [22]. Only in the
nized sediment or water, with the NO− 2 originating from the lowermost part of the NO− 3 zone was it at a limiting concen-
reduction of added NO− 3 [7,22,33,34]. The group of organ- tration. Therefore, in this sediment, NO− 3 reducers likely will
isms responsible for this reduction has not been identified, release NO− 2 for the anammox bacteria. Nitrite availability is
although denitrifying bacteria, and those bacteria promot- a prerequisite for anammox to be important in sediments in
ing the dissimilatory reduction of NO− +
3 to NH4 (DNRA), nature.
have NO− 2 as a free intermediate, and thus are likely can-
didates. The excretion of NO− 2 occurs, however, by many 3.3. Kinetics
bacteria which have the ability to reduce NO− 2 further. It
probably occurs when NO− 3 is at, or above, it’s limiting con- The affinity for NO−2 uptake by anammox bacteria and
centration and nitrate reducers can cover all their electron denitrifying bacteria has been investigated in homogenized
acceptor needs with the NO− 3 . This is always the case dur- sediment from the Skagerrak. The ratio of anammox to den-
15
ing the first part of most N-amendment experiments with itrification did not vary as a function of NO− 2 concentra-
relatively large additions of NO− 3 . In this case, nitrate is in tion [7], and it was concluded that the affinity for NO− 2
excess during the first part of the incubation, and a large por- uptake was similar in the 2 groups of bacteria. Furthermore,
tion of it is reduced only to NO− −
2 . The NO2 is then reduced the Km value for NO− 2 uptake was estimated to be below
460 T. Dalsgaard et al. / Research in Microbiology 156 (2005) 457–464
3 µM, possibly as low as 0.1 µM, for both processes in this
sediment where anammox accounted for approximately 65%
of the N2 production.
The Km value for NH+ 4 uptake by marine anammox bac-
teria is difficult to investigate in the type of experiments used
thus far to study sediment anammox. The problem is that
native NH+ 4 concentrations often are high, and NH4 pro-
+
duction during continued mineralization ensures that NH+
never reaches limiting levels. In Skagerrak sediments, in-
creasing the NH+ 4 concentration from 50 to 250 µM did not
change the anammox rate, indicating that the Km value was
well below 50 µM [7]. In natural sediments and anoxic wa-
ter columns, however, anammox may become NH+ 4 -limited.
In sediments anammox may be physically separated from
the major NH+ 4 source, namely the release of NH4 from
+ sediment, and the organic loading of the sediment is thus
anaerobic mineralization deeper in the system. In situations
like the one depicted in Fig. 1, NH+ 4 concentrations may be-
come very low in part of the NO− 3 zone and may thus limit
the anammox process. Alternatively, anammox bacteria may
have to compete for NH+ 4 with organisms conducting nitrifi-
cation or microphytobenthos (when light is available) assim-
ilating NH+ +
4 [22]. In anoxic water columns NH4 limitation
of anammox may occur as demonstrated in Golfo Dulce [6].
Here, NH+ 4 concentrations were close to the detection limit,
and the addition of 10 µM NH+ 4 stimulated the rate of anam-
mox 2- to 4-fold. Thus, the in situ NH+ 4 concentration was
at or below the Km value.
3.4. Effects of oxygen
For anammox bacteria isolated from waste water treat-
ment facilities, the anammox process appears to occur only
under strictly anaerobic conditions. As little as 1.1 µM O2
was sufficient to completely inhibit anammox activity in a
bioreactor [30]. The inhibition was reversible, and in exper-
iments with intermittent aeration, the rate of anammox was
the same before and after aeration [12]. There are no exist-
ing reports on the effects of O2 on anammox in the sea, but
it must be expected that marine anammox is also inhibited
by O2 . However, O2 concentrations likely fluctuate in the
NO− 3 -containing zones where marine anammox occurs and
as a result, marine anammox may have developed a higher
tolerance towards O2 .
4. Relative importance and regulation of marine
anammox
4.1. Marine sediments
The discovery of anammox in marine environments
means that denitrification is no longer synonymous with the
removal of fixed nitrogen. The deep sediments (700 m) of
the Skagerrak, which is part of the Danish belt seaway, sup-
port the maximum reported relative importance of anammox
in N2 production [10,33]. Here, anammox was responsible
for 67–79% of the total N2 production. Thus, in this sedi-
ment, anammox was more important than denitrification in
the removal of fixed nitrogen. However, the relative impor-
tance of anammox in nitrogen removal is highly variable,
and in some cases anammox activity has not been detected
(Fig. 2A). The relative importance of anammox in sediments
was originally correlated with sediment mineralization rate
4
and water depth [33]. Thus, in the initial reports of anammox
activity, anammox was responsible for 67, 24 and 2% of the
N2 production in the sediment of Skagerrak S9 (700 m),
Skagerrak S6 (380 m) and Aarhus Bay (16 m), respectively.
Water depth and sediment mineralization are generally re-
lated because with a deeper water column a larger fraction
of the organic matter is mineralized during transport to the
lower. Although anammox was relatively more important at
the deepest site of the Skagerrak, the absolute rates of anam-
mox were 3–4 times higher at the two shallower sites (see
Fig. 2).
This pattern was also observed in a study of anammox
and denitrification in 7 sediments [10] which ranged from
the deep Skagerrak with a very low sediment mineralization
rate, to the highly active sediment of Long Island Sound.
Overall, a good negative correlation was observed between
the relative importance of anammox and sediment mineral-
ization rate. Also, similar to the original observations, anam-
mox rates only varied by a factor of 6 between the stations,
whereas, denitrification rates varied by a factor of 114, and
was almost linearly dependent on the sediment mineraliza-
tion rate.
In the Thames estuary the relative importance of anam-
mox in N2 formation was linearly correlated with sediment
organic content [34]. Out of six stations, sampled along the
estuary from the discharge area of two very big sewage
treatment works in London, to the North Sea, only one site
deviated from this pattern. The organic content decreased
from the discharge area, where anammox was responsible
for 8% of the N2 production, towards the sea, where anam-
mox accounted for only 1% of the N2 production. Two
explanations were offered for this trend. Since anammox
was originally discovered in sewage treatment it was argued
that the Thames is seeded by anammox bacteria from the
sewage plant discharge. In this model the largest popula-
tion of anammox bacteria is located closest to the discharge.
Alternatively, the relationship between anammox and sed-
iment organic content may arise because a higher organic
content leads to a higher NO− 3 reduction which may lead
to a higher NO2 release and thus more NO−
−
2 available for
anammox [34].
In two other studies, the relative importance of anammox
was linked to the availability of NO− 3 . Risgaard-Petersen
et al. [22] compared 2 sites in 2 shallow Danish estuar-
ies (<1 m deep) and found that anammox accounted for
between 4 and 26% of the N2 production at the Randers
Fjord site but was undetectable in Norsminde

Fjord. They
attributed this difference to different NO− 3 availability in
 T. Dalsgaard et al. / Research in Microbiology 156 (2005) 457–464 461

Fig. 2. Relative anammox (anammox/total N2 production ×100%) (A) and absolute rates of anammox (B) and denitrification (C) measured in jar experiments
with homogenized sediment. Rates and relative anammox are estimated from 15 NO− 3 addition experiments. The water depth, station name and reference are
given on the x-axis. nd: not determined; bdl: below detection limit; ud: unpublished data. In Norsminde Fjord, Greece and Spain, only the relative contributions
of the 2 processes was measured.

the sediment. In Randers Fjord, NO− 3 in the water column microprofiles showed that there was no measurable NO− 3
and nitrification rates in the sediment are
high year round, in the anoxic zone of the sediment.
and microsensor profiles of porewater NO− 3 in the sedi- In continental shelf sediments from East and West Green-


ment showed that NO− 3
penetrated into the anoxic part of land, anammox was relatively important, ranging from 1 to
− 35% of the N2 production at 11 sites with water depths rang-
the sediment. In darkness, NO3 concentration decreased
from 23 µM at 1.4 mm depth (oxic–anoxic interface) to ing from 36 to 100 m [25]. In this study, area-based rates
zero at a depth of 3.7 mm. In the of anammox activity were estimated from whole core incu-

light, O2 penetrated to
a depth of 2.8 mm, where the NO− 3 concentration was
bations with 15 NO− 3 [23], and a striking linear correlation


20 µM, and NO− 3 penetrated to 5.5 mm. In Norsminde (R = 0.96) between bottom water NO−
2
3 concentrations and
Fjord, however, NO− 3 concentrations were low in the water area-based, absolute rates of anammox appeared. The NO− 3
column in the summer months. Also, the sediment was cov- concentrations in arctic bottom waters are rather stable over
ered with benthic microalgae
which, through active uptake, the year, so the NO− 3 concentration at the time of sampling
further suppressed sediment NO− 3 concentrations. Indeed, is a good indicator of the general NO− 3 availability in those
462 T. Dalsgaard et al. / Research in Microbiology 156 (2005) 457–464
sediments [25]. Also, since these sediments are below the
photic zone, there is no competition from microphytoben-
thos, as was the case in Norsminde Fjord [22]. Still, it is
surprising that the anammox rates correlated so closely with
the NO− 3 concentration in the bottom water. Sediment nitrifi-
cation was significant at all sites and the NO− 3 availability in
the sediment was therefore not governed by the bottom water
NO− 3 concentration alone [25]. The relation between rela-
tive anammox and sediment mineralization or water depth
was not investigated as the former was not measured and the
latter varied only between 36 and 100 m.
Can we find a common thread through these various ob-
servations? In Fig. 2 the published data on anammox in ma-
rine sediments have been ranked according to water depth,
and it appears that the relative importance of anammox in N2
production and water depth do correlate, as originally sug-
gested (Fig. 2A). The relationship is not perfect and other
environmental variables may also play a role in this regu-
lation. However, it is clear that the relative significance of
anammox in N2 production is highest at greater water depth.
Even though a high contribution of anammox in N2 forma-
tion can occasionally be found at shallow depth, anammox
is always relatively important for nitrogen removal at greater
depths. If this trend continues, anammox would be respon-
sible for at least 2/3 of the N2 production in sediments at
depths greater than those in Fig. 2.
The absolute rates of anammox, however, do not corre-
late with water depth, although there is a tendency towards
higher rates in shallower water (Fig. 2B). Denitrification
rates, on the other hand, do correlate with water depth, where
the highest rates are found at shallow water depths and rates
are always low for sediment in deep waters (Fig. 2C). In
the sediments so far studied denitrification rates vary over a
wider range (0.10–153 nmol cm−3 h−1 ) than anammox rates
(0.08–11 nmol cm−3 h−1 ) and denitrification is thus appar-
ently more responsive to organic carbon loading than anam-
mox. One might imagine that high rates of NH+ 4 release in
shallow water organic-rich sediments may stimulate anam-
mox, but this is apparently not the case. One reason for this
could be that anammox is already NH+ 4 -saturated in shallow
sediments. It is also possible that the higher electron donor
(organic matter) availability in organic rich sediments cre-
ates a higher demand for electron acceptor (i.e., NO− 2 and
NO− 3 ), and that a smaller fraction of the reduced NO −
3 is lib-
−
erated as NO2 . Anammox may thus not be able to keep up
with denitrification when electron donor availability is high.
These issues are obviously a ripe area for future research.
As suggested in some of the studies mentioned above,
anammox may also be regulated by the supply of NO− 3 from
both bottom waters and from nitrification. Higher NO− 3 con-
centration in the bottom water or higher nitrification rates
will result in a deeper penetration of NO− 3 into the sediment.
Consequently, the zone where denitrification (and nitrate re-
duction to ammonia) will be NO− 3 saturated, and where NO2
−
therefore may be excreted to the porewater (see above), will
be thicker and anammox may become more important. In
situations where denitrification is electron-donor limited, an
increase in the availability of organic matter would stimulate
denitrification. The penetration of NO− 3 into the sediment
would thus decrease and the thickness of the zone in which
NO− 2 would be released, and available for anammox, would
also decrease. Assuming that the rate of anammox per vol-
ume of sediment would remain constant, a thinner zone of
anammox would give a lower overall rate of anammox in
the sediment, and an even lower contribution of anammox to
N2 production.
Another important variable regulating the importance of
anammox may be environmental stability. If anammox bac-
teria in nature grow as slowly as we believe (see above), then
the anammox process is probably only significant in stable
environments where there is a prolonged time for the bac-
terial population to develop. Denitrifiers have much higher
growth rates which gives them a competitive advantage over
the anammox bacteria in fluctuating environments.
Clearly, the factors controlling the environmental signif-
icance of anammox are still poorly understood. The depth
dependence in Fig. 2 may indicate that water depth, and
therefore sediment mineralization rate, is the most important
driver controlling the relative importance of anammox in N2
production. Other factors, then, such as NO− 3 concentration,
microphytobenthos activity, seeding of anammox bacteria,
or local gradients in organic content may provide secondary,
though still important, control.
4.2. Anoxic water columns and sea ice
The anammox process is also important in N2 produc-
tion in anoxic water bodies. In Golfo Dulce, a restricted
basin on the Pacific coast of Costa Rica, anammox was
found to account for 19–35% of the N2 formation in the
anoxic bottom waters [6]. Nitrate-rich surface water from
the Pacific mixes into the anoxic bottom water of the basin,
and the NH+ 4 produced by the mineralization of sinking
organic matter in the water column was immediately con-
sumed by anammox bacteria. As a result, the anoxic water
column was almost free of NH+ +
4 , and the low NH4 concen-
trations limited the rate of anammox. In the anoxic water
column denitrification was the main mineralization process,
and the relative importance of anammox was thus deter-
mined by the relationship between NO− 3 consumption and
NH+ 4 production by the denitrifying community. Theoreti-
cally, anammox should be responsible for 29% of the N2
production provided all NH+ 4 production coupled to den-
itrification was oxidized by anammox [6]. However, this
theoretical limit may be exceeded because of preferential
degradation of nitrogen-rich organic matter by denitrifica-
tion [19,37]. In the Black Sea, anammox was shown to occur
in the anoxic, non-sulfidic, zone where the NH+ 4 from the
−
anoxic bottom water meets the NO3 from the surface waters
[15]. Thus, in this environment anammox is not closely cou-
pled to denitrification and other stoichiometric constraints
may apply.
 T. Dalsgaard et al. / Research in Microbiology 156 (2005) 457–464
The anammox process has also been found active in arctic
sea ice [24]. The process was not detectable in ice less than
1 year old, but it was found to account for up to 19% of
the N2 production in individual layers of a several years old
ice floe. However, anammox was only important in a thin
horizon and was thus only responsible for 0–5% of the total
nitrogen removal in the ice. It is argued that the anammox
process, due to the very slow growth of the bacteria [13],
requires stable conditions and a long time to develop, and
this is why it was not present in the first year ice.
5. Anammox and the marine nitrogen budget
The removal of fixed nitrogen from the marine environ-
ment is believed to occur predominantly in anoxic sediments
and in suboxic waters including the oxygen minimum zones
of the Arabian Sea and the north and south eastern equatorial
Pacific Ocean. By current estimates, sediments account for
about 2/3 of the total N2 production, and the water column
is responsible for the remaining 1/3 [4]. Sediments on the
continental slope (>150 m depth), and in the deep sea are
assumed to be responsible for 53% of the sediment N2 pro-
duction [16]. If anammox is responsible for more than half
of this (as current results thus far indicate), and furthermore,
is responsible for a significant part of the nitrogen removed
at shallower depths, this “new” process may be expected to
carry out 1/3–2/3 of the global sedimentary nitrogen re-
moval. Considering that about 87% of the ocean is deeper
than 1000 m and the average depth of the oceans is 3800 m
[31] it may be expected that anammox will be the dominant
nitrogen removal process in most sediments. Furthermore,
if anammox is as important in the oxygen minimum zones
of the oceans as in Golfo Dulce, it may be expected to
account for 35% of the marine water column nitrogen re-
moval. Anammox may thus be responsible for 1/3–1/2 of
the global removal of fixed nitrogen from the marine en-
vironment [9]. While a very coarse estimate, this number
emphasizes the need for a more detailed mapping of anam-
mox in both benthic and pelagic environments.
6. Conclusions
As shown above, our knowledge of marine anammox bac-
teria, and the magnitude and controls of the process, has
evolved greatly in recent years, but many questions remain.
Indeed, much of our understanding is borrowed from studies
in waste water treatment systems. The overall stoichiome-
try of the anammox process in the marine realm appears
the same as in waste water treatment systems, and closely
related bacteria belonging to the anammox branch of the
Planctomycetales have been identified in these two environ-
ments. The importance of anammox in removing nitrogen in
sediments seems to be highest at greater water depths where
sediment mineralization rates are lower. In shallower waters,
463
increases in sediment organic loading apparently augment
denitrification much more than anammox, and the latter is
of much lower importance. It is suggested that other vari-
ables such as NO− 3 availability, local gradients in sediment
organic content, or the presence of microphytobenthos, may
regulate anammox within the overall frame defined by the
organic loading of the sediment. The anammox process is
also very important in anoxic water columns where it may
account for 35% of the nitrogen removal. It is argued that
anammox in sediments and anoxic water bodies may amount
to 1/3–1/2 of the global marine nitrogen removal.
Acknowledgements
T.D. was supported by the Swedish Foundation for Strate-
gic Marine Research (MISTRA, Dnr 2001-108) and by the
EU research project MedVeg (QLRT-2000-02456). We also
acknowledge Financial support for field work from the Dan-
ish Research Council. D.E.C. and B.T. were further sup-
ported by the Danish National Research Foundation.
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