bs_bs_banner

Environmental Microbiology Reports (2012)

doi:10.1111/1758-2229.12000

Prismatic magnetite magnetosomes from cultivated

Magnetovibrio blakemorei strain MV-1: a magnetic
fingerprint in marine sediments?
Luigi Jovane,1* Fabio Florindo,2
Dennis A. Bazylinski3 and Ulysses Lins4
1
Instituto Oceanogrfico, Universidade de So Paulo,
So Paulo, Brazil.
2
Istituto Nazionale di Geofisica e Vulcanologia, Rome,
Italy.
3
School of Life Sciences, University of Nevada at Las
Vegas, Las Vegas, Nevada, USA.
4
Instituto de Microbiologia, Universidade Federal do Rio
de Janeiro, Rio de Janeiro, Brazil.
Summary
The magnetic properties (first-order reversal curves,
ferromagnetic resonance and decomposition of saturation remanent magnetization acquisition) of Magnetovibrio blakemorei, a cultivated marine magnetotactic
bacterium, differ from those of other magnetotactic
species from sediments deposited in lakes and marine
habitats previously studied. This finding suggests that
magnetite produced by some magnetotactic bacteria
retains magnetic properties in relation to the crystallographic structure of the magnetic phase produced
and thus might represent a magnetic fingerprint for a
specific magnetotactic bacterium. The use of this fingerprint is a non-destructive, new technology that
might allow for the identification and presence of specific species or types of magnetotactic bacteria in
certain environments such as sediments.
Introduction
Magnetotactic bacteria are prokaryotic microorganisms
which internally biomineralize magnetite (Fe3O4) or greigite
(Fe3S4) crystals (nanoparticles) or both enveloped in a lipid
bilayer membrane. These organelles, called magnetosomes, contribute significantly to the magnetic properties
of sediments (Bazylinski and Frankel, 2004). Magnetosomes have very interesting biotechnological characteristics because they are made of singular, perfect,
single-magnetic-domain (SD) crystals of magnetite or
Received 7 June, 2012; revised 19 September, 2012; accepted 23
September, 2012. *For correspondence. E-mail jovane@usp.br; Tel.
(+55) 11 31096609; Fax (+55) 11 31096610.

2012 Society for Applied Microbiology and Blackwell Publishing Ltd

greigite that are usually aligned in chains within the cell

(Schler, 2006).
Here we present the results of a study using Magnetovibrio blakemorei strain MV-1: a cultivated magnetotactic marine vibrio that produces prismatic magnetite
magnetosomes (~50100 nm diameter) and was originally isolated from sediment collected from a salt marsh
pool near the Neponset River (Milton, MA, USA) (Bazylinski et al., 1988; 2012). In this report, we focus on the
magnetosome magnetite crystals of M. blakemorei and
their importance in future studies of sediments and their
environmental significance in oceanographic changes.
Because M. blakemorei lives naturally in the sediment
and water column and produces pseudo-hexagonal prismatic magnetite magnetosomes under microaerobic and
strictly anaerobic conditions, its magnetosome crystals
may provide a magnetic fingerprint of anaerobic conditions that can be identified in the geological record.
The presence of magnetotactic bacteria in sediments,
detected as magnetosome crystals, namely magnetofossils, may be the primary source of natural remanence in
these sediments and is able to maintain the noninteracting SD bulk magnetic properties because of the
preservation of the chain structure (Moskowitz et al.,
1993). Recently, their presence has been associated to
hyperthermal events and high-productivity climatic events
(Schumann et al., 2008; Roberts et al., 2011; Chang
et al., 2012) outlining the importance of magnetotactic
bacteria as a proxy for reconstructing paleoenvironments.
Magnetic analyses provide a quick and efficient tool to
determine the presence of magnetotactic bacteria and/or
magnetosome crystals in recent or ancient sediments or
in the water columns of aquatic habitats (Roberts et al.,
2011; 2012). In particular, first-order reversal curves
(FORC) (Roberts et al., 2000; Egli et al., 2010), ferromagnetic resonance (FMR) measurements (e.g. Weiss et al.,
2004) and decomposition of saturation remanent magnetization (Mr) acquisition (DAM) (Heslop et al., 2002) are
well-understood, rapid, inexpensive and non-destructive
measurements which can be used to determine magnetic
properties, magnetic grain-size distributions, interactions
and crystallographic anomalies. FORC diagrams are
contour plots of the second derivative of magnetic fields
Hu and Hc, which are related to the coercivity (Hu) and bias

L. Jovane, F. Florindo, D. A. Bazylinski and U. Lins

or interactions (Hc) measured in a series of partial hysteresis curves. FMR and acquisition of magnetization
saturation are the measurements of the absorption of
magnetic anisotropy and the magnetic moment while a
magnetic field is applied respectively.
The presence of magnetofossils is much more common
than has been reported in the literature (Roberts et al.,
2012) and their abundance in marine sediments is related
to concentration of organic matter in sediments, and, most
importantly, is directly related to phases of high primary
productivity in oceans (Schumann et al., 2008; Roberts
et al., 2011; Chang et al., 2012). Magnetofossils may be
the main driver of magnetization in marine sediments. If
we are able to discern and recognize different environmental conditions and ecosystems from the magnetic fingerprints left by magnetotactic bacteria in the sedimentary
record, it provides a further and substantial leap forward in
terms of studying the geological record (Yamazaki and
Kawahata, 1998; Housen and Moskowitz, 2006; Schumann et al., 2008; Roberts et al., 2011).
Results and discussion
Cells of M. blakemorei were cultivated in anaerobic liquid
medium as previously described (Dean and Bazylinski,
1999). Transmission electron microscope images of a
single magnetosome and a cell of M. blakemorei are
shown in Fig. 1A and B respectively. The magnetosome
membrane surrounding the magnetite crystal is clearly
evident (Fig. 1A). The FORC diagram of M. blakemorei
cells (Fig. 1C) is a horizontal ridge beginning at ~ 30 mT
with its maximum peak above 50 mT on the Hc axis, but is
only a few mT above 0 on the Hu axis. The FORC diagram
is slightly wider around 40 mT and narrower at 60 mT.
FORC diagrams of magnetosomes (e.g. Pan et al., 2005;
Fischer et al., 2008) show a horizontal central ridge developed from the origin and highest peak below 40 mT.
These specifications demonstrate that the FORC diagram
of MV-1 we identified differ from any other FORC of magnetosomes, and are directly related to the crystallographic
shape of the magnetosomes produced by this bacterium.
Although FMR (Fig. 1D) and DAM (Fig. 1E and F) measurements of M. blakemorei are very similar to those of
Weiss and colleagues (2004) and Roberts and colleagues
(2011), respectively, they show slight differences that are
directly related to the singular shape of the magnetic
crystals. Interestingly, the FORC diagram for M. blakemorei is significantly different from those of other magnetotactic bacteria, magnetosomes and magnetofossils.
We must consider that sediments recovered from environmental samples typically retain a much lower concentration of bacteria (magnetosomes) as unique species or
mixtures of species (e.g. Kopp et al., 2006; Yamazaki,
2008; 2009; Chang et al., 2012); however, the interaction/

non-interaction ratio, FMR, DAM and other methods

should be able to distinguish the various shapes of magnetosomes produced by different magnetotactic bacteria.
Nevertheless, the magnetosomes might be subjected to
reducing diagenetic processes that corrode partially the
original shape or dissolve completely selected species
(Yamazaki and Kawahata, 1998).
Each species of magnetotactic bacteria biomineralizes
magnetic particles with a singular crystal composition
(e.g. Faivre et al., 2008; Popa et al., 2009) which may
relate to external conditions thereby producing magnetosome crystals (Yamazaki and Kawahata, 1998) with magnetic properties that can be often distinguished in FORC
diagrams. Common FORCs of magnetofossils (Yamazaki,
2008; 2009; Abrajevitch and Kodama, 2009; 2011; Kind
et al., 2011; Roberts et al., 2011; 2012; Yokoyama et al.,
2011) show, like most magnetosomes, a horizontal ridge
initiating at the origin with the main peak between 20 and
40 mT.
First-order reversal curve results on cultured magnetotactic bacteria have been published (e.g. Fischer et al.,
2008), and most of these involve magnetotactic bacteria
that are not marine. In this case the species belong to the
genus Magnetospirillum which produces cuboctahedral
magnetosomes instead of the prismatic ones produced by
M. blakemorei. The FORC diagram for M. blakemorei is
significantly different; consequently, we assume that the
magnetic characteristics observed here are directly associated with this species and the environment in which it
lives and represent the fingerprint needed to detect the
presence of this magnetotactic bacterium and its magnetofossils in sediments.

Conclusions
We show a series of specific non-destructive techniques
to determine the magnetic properties of a cultivated
marine magnetotactic bacterium originally recovered from
sediments (e.g. Egli, 2004) in this case the bacterium:
M. blakemorei. We show that the magnetic properties of
this bacterium, a species that lives in marine sedimentary
ecosystems, are different from those of other marine and
freshwater magnetotactic bacteria and thus could be used
as a magnetic fingerprint for the presence of M. blakemorei in natural habitats such as sediments. The nondestructive technique presented here allows for an
alternative microbiological method of studying magnetotactic bacteria in their natural habitats.

Acknowledgements
We thank Evandro Luz Duarte and Leandro Barbosa of Instituto de Fsica da Universidade de So Paulo helping in FMR
measurements. D. A. B. is supported by US National Science

2012 Society for Applied Microbiology and Blackwell Publishing Ltd, Environmental Microbiology Reports

 2012 Society for Applied Microbiology and Blackwell Publishing Ltd, Environmental Microbiology Reports

1m

Derivate of absorption

-8000

-6000

-4000

-2000

2000

Sample

geff

80

246.6

160

240
320
400
Magnetic Field (mT)

Value (G)

480

560

421.9

Bhigh
(mT)

Blow
(mT)

20

Ferromagnetic Resonance (FMR)

Ca.M.b. 1.802

Field increment = 0.5 mT

Smoothing Factor = 5

-50
150

50

100

150

200

250

300

64.51

138.5

Ms
(Am2)

Mr
(Am2)

100

200

400

600

800

31.6 mT

50
100
Magnetic Field (mT)

50.7 mT

gradient
grad-synt1 (89%)
grad-synt2 (11%)
grad-synt-sum

Decomposition of Mr acquisition (DAM)

43.53

35.71

350

Hcr
(mT)

Hc

80

(mT)

-10
0

50
100
Magnetic Field (mT)

Mr
synt1 (Mr=58 mT,
log (B1/2)=1.71,
DP=0.09)
synt2 (Mr=7 mT,
log(B1/2)=1.5,
DP=0.11)
synt-sum

0.251

175.4

(mT)

BFWHM

60

Mr acquisition

386.7

(mT)

Beff

Hc (mT)

10

20

30

40

50

60

70

35.1

140.3

Bhigh
(mT)

Blow
(mT)

40

First order reversal curve (FORC)

150

Fig. 1. A. Transmission electron microscope (TEM) image of a single magnetosome of Magnetovibrio blakemorei. This magnetosome contains a pseudo-hexagonal prismatic crystal of
magnetite surrounded by the magnetosome membrane (arrow).
B. TEM image of an unstained, whole cell illustrating a magnetosome chain traversing the cell along its long axis.
C. First-order reversal curve (FORC) diagram of cells of M. blakemorei strain MV-1, which is a horizontal ridge beginning at ~ 30 mT with its maximum peak above 50 mT on the Hc axis, but
is only a few mT above 0 on the Hu axis. The FORC diagram is slightly wider around 40 mT and narrower at 60 mT.
D. Ferromagnetic resonance (FMR) shows a curve with three peaks at 180, 320 and 370 mT, and a shape similar to that of Weiss and colleagues (2004) for the same organism.
E. Saturation remanent magnetization (Mr) acquisition, and (F) decomposition of Mr (DAM) which show a main and narrow component at 50 mT and a minor and wider at ~ 30 mT. The table
shows main FMR and hysteresis magnetic parameters. TEM images were produced with a FEI Morgagni TEM. FMR was run at Instituto de Fsica da Universidade de So Paulo (Brazil) with
X-band Bruker EMX microspectrometer emitting a microwave frequency of ~ 9.4 GHz, microwave power of ~ 0.632 mW, and integrated over three magnetic field sweeps from 0 to 700 mT.
FORC smoothing parameters are shown in the left corner. DAM components parameters are shown on the lower right corner of the figure. Hysteresis parameters, FORC and DAM were
performed at the Istituto Nazionale di Geofisica e Vulcanologia (Italy) with a vibrating sample magnetometer (VSM, Princeton Measurements Corporation Model 3900 Micromag).

-15

-10

-5

10

15

Hu (mT)

-6

Magnetization remanence (Mr) (Am2)

x10

gradient

Transmission electron microscope (TEM)

Magnetotactic bacteria fingerprint in sediments

3

L. Jovane, F. Florindo, D. A. Bazylinski and U. Lins

Foundation (NSF) grant EAR-0920718. Financial support

from Brazilian CNPq, FAPERJ and CAPES is acknowledged
by U. L. The authors wish to thank Ken Timmis (Editor) and
two anonymous reviewers for comments that helped to
improve this paper.

Author contribution
All authors contributed to acquisition, analysis and interpretation of data presented in this paper. L. J.: Overall
coordination of writing. D. A. B.: Sediment sampling.
U. L.: Bacteria cultivation. L. J. and F. F.: Magnetic properties and environmental interpretation.
References
Abrajevitch, A., and Kodama, K. (2009) Biochemical vs. detrital mechanism of remanence acquisition in marine carbonates: a lesson from the K-T boundary interval. Earth Planet
Sci Lett 286: 269277. doi: 10.1016/j.epsl.2009.06.035.
Abrajevitch, A., and Kodama, K. (2011) Diagenetic sensitivity
of paleoenvironmental proxies: a rock magnetic study
of Australian continental margin sediments. Geochem
Geophys Geosyst 12: Q05Z24. doi: 10.1029/
2010GC003481.
Bazylinski, D.A., and Frankel, R.B. (2004) Magnetosome formation in prokaryotes. Nat Rev Microbiol 2: 217230. doi:
10.1038/nrmicro842.
Bazylinski, D.A., Frankel, R.B., and Jannasch, H.W. (1988)
Anaerobic magnetite production by a marine magnetotactic
bacterium. Nature 334: 518519.
Bazylinski, D.A., Williams, T.J., Lefvre, C.T., Trubitsyn, D.,
Fang, J., Beveridge, T.J., et al. (2012) Magnetovibrio
blakemorei, gen. nov. sp. nov., a new magnetotactic bacterium (Alphaproteobacteria: Rhodospirillaceae) isolated
from a salt marsh. Int J Syst Evol Microbiol. In press, online
doi: 10.1099/ijs.0.044453-0.
Chang, L., Roberts, A.P., Williams, W., Fitz Gerald, J.D.,
Larrasoaa, J.C., Jovane, L., and Muxworthy, A.R.
(2012) Giant magnetofossils and hyperthermal events.
Earth Planet Sci Lett 351352: 258269. doi: 10.1016/
j.epsl.2012.07.031.
Dean, A.J., and Bazylinski, D.A. (1999) Genome analysis of
several magnetotactic bacterial strains using pulsed-field
gel electrophoresis. Curr Microbiol 39: 219225.
Egli, R. (2004) Characterization of individual rock magnetic
components by analysis of remanence curves: 1. Unmixing
natural sediments. Stud Geophys Geod 48: 391446. doi:
10.1023/B:SGEG.0000020839.45304.6d.
Egli, R., Chen, A.P., Winklhofer, M., Kodama, K.P., and
Horng, C.S. (2010) Detection of noninteracting single
domain particles using first-order reversal curve diagrams.
Geochem Geophys Geosyst 11: Q01Z11. doi: 10.1029/
2009GC002916.
Faivre, D., Menguy, N., Psfai, M., and Schler, D. (2008)
Environmental parameters affect the physical properties of
fast-growing magnetosomes. Am Mineral 93: 463469.
Fischer, H., Mastrogiacomo, G., Lffler, J.F., Warthmann,
R.J., Weidler, P.G., and Gehring, A.U. (2008) Ferromagnetic resonance and magnetic characteristics of intact

magnetosome chains in Magnetospirillum gryphiswaldense. Earth Planet Sci Lett 270: 200208. doi: 10.1016/
j.epsl.2008.03.022.
Heslop, D., Dekkers, M.J., Kruiver, P.P., and van Oorschot,
I.H.M. (2002) Analysis of isothermal remanent magnetisation acquisition curves using the expectation-maximization
algorithm. Geophys J Int 148: 5864.
Housen, B.A., and Moskowitz, B.M. (2006) Depth distribution
of magnetofossils in near surface sediments from the
Blake/Bahama Outer Ridge, western North Atlantic Ocean,
determined by low-temperature magnetism. J Geophys
Res 111: G01005.
Kind, J., Gehring, A.U., Winklhofer, M., and Hirt, A.M. (2011)
Combined use of magnetometry and spectroscopy
for identifying magnetofossils in sediments. Geochem
Geophys Geosyst 12: Q08008.
Kopp, R.E., Nash, C.Z., Kobayashi, A., Weiss, B.P., Bazylinski, D.A., and Kirschvink, J.L. (2006) Ferromagnetic resonance spectroscopy for assessment of magnetic
anisotropy and magnetostatic interactions: a case study of
mutant magnetotactic bacteria. J Geophys Res 111:
B12S25. doi: 10.1029/2006JB004529.
Moskowitz, B.M., Frankel, R.B., and Bazylinski, D.A. (1993)
Rock magnetic criteria for the detection of biogenic magnetite. Earth Planet Sci Lett 120: 283300. doi: 10.1016/
0012-821X(93)90245-5.
Pan, Y.X., Petersen, N., Winklhofer, M., Davila, A.F., Liu,
Q.S., Frederichs, T., et al. (2005) Rock magnetic properties
of uncultured magnetotactic bacteria. Earth Planet Sci Lett
237: 311325. doi: 10.1016/j.epsl.2005.06.029.
Popa, R., Fang, W., Nealson, K.H., Souza-Egipsy, V.,
Berquo, T.S., Banerjee, S.K., and Penn, L.R. (2009) Effect
of oxidative stress on the growth of magnetic particles in
Magnetospirillum magneticum. Int Microbiol 12: 4957.
Roberts, A.P., Pike, C.R., and Verosub, K.L. (2000) FORC
diagrams: a new tool for characterizing the magnetic properties of natural samples. J Geophys Res 105: 28461
28475.
Roberts, A.P., Florindo, F., Villa, G., Chang, L., Jovane, L.,
Bohaty, S.M., et al. (2011) Magnetotactic bacterial abundance in pelagic marine environments is limited by organic
carbon flux and availability of dissolved iron. Earth Planet
Sci Lett 310: 441452.
Roberts, A.P., Chang, L., Heslop, D., Florindo, F., and
Larrasoaa, J.C. (2012) Searching for single domain magnetite in the pseudo-single-domain sedimentary haystack:
implications of biogenic magnetite preservation for sediment magnetism and relative paleointensity determinations. J Geophys Res 117: B08104. doi: 10.1029/
2012JB009412.
Schler, D. (2006) Magnetoreception and Magnetosomes in
Bacteria Microbiological Monographs. New York, NY,
USA: Springer. 3, p. 90.
Schumann, D., Raub, T.D., Kopp, R.E., Guerquin-Kerne, J.L.,
Wu, T.D., Rouiller, I., et al. (2008) Gigantism in unique
biogenic magnetite at the PaleoceneEocene Thermal
Maximum. Proc Natl Acad Sci USA 105: 1764817653.
Weiss, B.P., Kim, S.S., Kirschvink, J.L., Kopp, R.E., Sankaran, M., Kobayashi, A., and Komeili, A. (2004) Ferromagnetic resonance and low temperature magnetic tests for
biogenic magnetite. Earth Planet Sci Lett 224: 7389.

2012 Society for Applied Microbiology and Blackwell Publishing Ltd, Environmental Microbiology Reports

Magnetotactic bacteria fingerprint in sediments

Yamazaki, T. (2008) Magnetostatic inter actions in deep-sea
sediments inferred from first-order reversal curve diagrams: implications for relative paleointensity normalization. Geochem Geophys Geosyst 9: Q02005. doi: 10.1029/
2007GC001797.
Yamazaki, T. (2009) Environmental magnetism of Pleistocene sediments in the North Pacific and Ontong-Java
Plateau: temporal variations of detrital and biogenic components. Geochem Geophys Geosyst 10: Q07Z04. doi:
10.1029/2009GC002413.

Yamazaki, T., and Kawahata, H. (1998) Organic carbon flux

controls the morphology of magnetofossils in marine sediments. Geology 26: 10641066.
Yokoyama, Y., Webster, J.M., Cotterill, C., Braga, J.C.,
Jovane, L., Mills, H., et al., and the IODP 325 Scientists
(2011) Great Barrier Reefs reveals past sea-level, climate
and environmental changes during the end of the last ice
age. Sci Drill 12: 3245.

2012 Society for Applied Microbiology and Blackwell Publishing Ltd, Environmental Microbiology Reports