Abstracts
Analyses of 16S rDNA genes were used to identify the microbiota isolated from the mucus of the zoanthid Palythoa caribaeorum at Porto de Galinhas on the coast of Pernambuco State, Brazil. This study is important as the first report of this association, because of the potential biotechnological applications of the bacterium Alcanivorax dieselolei, and as evidence for the presence of a hydrocarbon degrading bacterium in a reef ecosystem such as Porto de Galinhas.
16S rDNA; microbiota; biodegradation; hydrocarbons; cnidarians
Análises dos genes 16S rDNA foram empregadas para identificar a microbiota isolada do muco do zoantídeo Palythoa caribaeorum de Porto de Galinhas, litoral do estado de Pernambuco, Brasil. Este estudo é importante pelo ineditismo dessa associação, pelas relevantes aplicações biotecnológicas da bactéria Alcanivorax dieselolei e pela indicação da presença de uma bactéria degradadora de hidrocarbonetos em um ecossistema recifal como o de Porto de Galinhas.
Palvras-chave: 16S rDNA; microbiota; biodegradação; hidrocarbonetos; cnidários
1 Introduction
Coral reefs are complex ecosystems that provide microniches for enormous diversities
of microorganisms (Ainsworth et al., 2010Ainsworth, TD., Thurber, RV. and Gates, RD., 2010. The future of
coral reefs: a microbial perspective. Trends in Ecology & Evolution, vol.
25, no. 4, p. 233-240. http://dx.doi.org/10.1016/j.tree.2009.11.001.
PMid:20006405
http://dx.doi.org/10.1016/j.tree.2009.11...
)
associated with marine invertebrates such as sponges, cnidarians, and mollusks
(Sfanos et al., 2005Sfanos, K., Harmody, D., Dang, P., Ledger, A., Pomponi, S.,
McCarthy, P. and Lopez, J., 2005. A molecular systematic survey of cultured
microbial associates of deep-water marine invertebrates. Systematic and Applied
Microbiology, vol. 28, no. 3, p. 242-264.
http://dx.doi.org/10.1016/j.syapm.2004.12.002. PMid:15900971
http://dx.doi.org/10.1016/j.syapm.2004.1...
). Cnidarians,
especially corals, have many microorganisms associated with their tissues and mucus
(Rohwer et al., 2002Rohwer, F., Seguritan, V., Azam, F. and Knowlton, N., 2002.
Diversity and distribution of coral associated bacteria. Marine Ecology Progress
Series, vol. 243, p. 1-10.
http://dx.doi.org/10.3354/meps243001.
http://dx.doi.org/10.3354/meps243001...
; Chimetto et al., 2009Chimetto, LA., Brocchi, M., Gondo, M., Thompson, CC., Gomez-Gil, B.
and Thompson, FL., 2009. Genomic diversity of vibrios associated with the
Brazilian coral Mussismilia hispida and its sympatric zoanthids
(Palythoa caribaeorum, Palythoa variabilis
and ). Zoanthus solanderiJournal of Applied Microbiology, vol.
106, no. 6, p. 1818-1826. http://dx.doi.org/10.1111/j.1365-2672.2009.04149.x.
PMid:19291243
http://dx.doi.org/10.1111/j.1365-2672.20...
; Castro et al., 2010Castro, AP., Araújo JUNIOR, SD., Reis, AMM., Moura, RL.,
Francini-Filho, RB., Pappas JUNIOR, G., Rodrigues, TB., Thompson, FL. and
Krüger, RH., 2010. Bacterial community associated with healthy and diseased reef
coral Mussismilia hispida from eastern Brazil. Microbial Ecology, vol. 59, no.
4, p. 658-667. http://dx.doi.org/10.1007/s00248-010-9646-1.
PMid:20352207
http://dx.doi.org/10.1007/s00248-010-964...
). Coral mucus – a micro-layer of
polysaccharides and glycoproteins deposited over the surfaces of these animals – has
been found to contain representatives of the three primary domains Archaea,
Eubacteria and Eukarya (Meikle et al., 1988Meikle, P., RICHARDS, GN. and YELLOWLEES, D., 1988. Structural
investigations on the mucus from six species of coral. Marine Biology, vol. 99,
no. 2, p. 187-193. http://dx.doi.org/10.1007/BF00391980.
http://dx.doi.org/10.1007/BF00391980...
;
Wegley et al., 2007Wegley, L., Edwards, R., Rodriguez-Brito, B., Liu, H. and Rohwer,
F., 2007. Metagenomic analysis of the microbial community associated with the
coral Porites astreoides. Environmental Microbiology, vol. 9, no. 11, p.
2707-2719. http://dx.doi.org/10.1111/j.1462-2920.2007.01383.x.
PMid:17922755
http://dx.doi.org/10.1111/j.1462-2920.20...
).
While a number of workers studies have examined the microbiota associated with corals
and other marine invertebrates (Castro et al.,
2010Castro, AP., Araújo JUNIOR, SD., Reis, AMM., Moura, RL.,
Francini-Filho, RB., Pappas JUNIOR, G., Rodrigues, TB., Thompson, FL. and
Krüger, RH., 2010. Bacterial community associated with healthy and diseased reef
coral Mussismilia hispida from eastern Brazil. Microbial Ecology, vol. 59, no.
4, p. 658-667. http://dx.doi.org/10.1007/s00248-010-9646-1.
PMid:20352207
http://dx.doi.org/10.1007/s00248-010-964...
; Menezes et al., 2010Menezes, CBA., Bonugli-Santos, RC., Miqueletto, PB., Passarini,
MRZ., Silva, CHD., Justo, MR., Leal, RR., Fantinatti-Garboggini, F., Oliveira,
VM., Berlinck, RGS. and Sette, LD., 2010. Microbial diversity associated with
algae, ascidians and sponges from the north coast of São Paulo state, Brazil.
Microbiological Research, vol. 165, no. 6, p. 466-482.
http://dx.doi.org/10.1016/j.micres.2009.09.005. PMid:19879115
http://dx.doi.org/10.1016/j.micres.2009....
; Trindade-Silva et al., 2012Trindade-Silva, AE., Rua, C., Silva, GGZ., Dutilh, BE., Moreira,
APB., Edwards, RA., Hajdu, E., Lobo-Hajdu, G., Vasconcelos, AT., Berlinck, RGS.
and Thompson, FL., 2012. Taxonomic and functional microbial signatures of the
endemic marine sponge Arenosclera brasiliensis. PLoS ONE, vol. 7, no. 7, p.
e39905. http://dx.doi.org/10.1371/journal.pone.0039905.
PMid:22768320
http://dx.doi.org/10.1371/journal.pone.0...
), zoanthids have
been little examined in that respect, although some investigations of the diversity
of bacteria found on zoanthids by Chimetto et al.
(2008Chimetto, LA., Brocchi, M., Thompson, CC., Martins, RCR., Ramos, HR.
and Thompson, FL., 2008. Vibrios dominate as culturable nitrogen-fixing bacteria
of the Brazilian coral . Mussismilia hispidaSystematic and
Applied Microbiology, vol. 31, no. 4, p. 312-319.
http://dx.doi.org/10.1016/j.syapm.2008.06.001. PMid:18678453
http://dx.doi.org/10.1016/j.syapm.2008.0...
, 2009Chimetto, LA., Brocchi, M., Gondo, M., Thompson, CC., Gomez-Gil, B.
and Thompson, FL., 2009. Genomic diversity of vibrios associated with the
Brazilian coral Mussismilia hispida and its sympatric zoanthids
(Palythoa caribaeorum, Palythoa variabilis
and ). Zoanthus solanderiJournal of Applied Microbiology, vol.
106, no. 6, p. 1818-1826. http://dx.doi.org/10.1111/j.1365-2672.2009.04149.x.
PMid:19291243
http://dx.doi.org/10.1111/j.1365-2672.20...
, 2011Chimetto, LA., Cleenwerck, I., Alves JUNIOR, N., Silva, BS.,
Brocchi, M., Willems, A., De Vos, P. and Thompson, FL., 2011. Vibrio communis
sp. nov., isolated from the marine animals Mussismilia hispida,
, . Phyllogorgia dilatataPalythoa caribaeorum,
Palythoa variabilis and Litopenaeus
vannameiInternational Journal of Systematic and Evolutionary
Microbiology, vol. 61, no. 2, p. 362-368.
http://dx.doi.org/10.1099/ijs.0.019729-0. PMid:20305064
http://dx.doi.org/10.1099/ijs.0.019729-0...
) identified 16S rDNA sequences of
bacteria of the genus Vibrio.
The zoanthid Palythoa caribaeorum (Duchassaing and Michelotti, 1860)
is a cnidarian commonly found on reefs in the Caribbean region and Brazil (Mueller and Haywick, 1995MUELLER, E. and HAYWICK, DW., 1995. Sediment assimilation and
calcification by the Western Atlantic reef zoanthid, Palythoa
caribaeorum.Bulletin de l'Institut Oceanographique, vol. 14, p.
89-100.). It can form dense
layers on reefs as it is a strong competitor for space (Pérez et al., 2005PÉREZ, CD., VILA-NOVA, DA. and SANTOS, AM., 2005. Associated
community with the zoanthid (Duchassaing & Michelotti, 1860) (Cnidaria,
Anthozoa) from littoral of Pernambuco, Brazil. Palythoa
caribaeorumHydrobiologia, vol. 548, no. 1, p. 207-215.
http://dx.doi.org/10.1007/s10750-005-5441-2.
http://dx.doi.org/10.1007/s10750-005-544...
), is tolerant of environmental stresses
(Sebens, 1982SEBENS, KP, 1982. Intertidal distribution of zoanthids on the
Caribbean coast of Panama: effects of predation and dessication. Bulletin of
Marine Science, vol. 32, p. 316-335.), demonstrates a high
reproductive capacity (Acosta and Asbahr,
2000Acosta, A. and Asbahr, M., 2000. Reproductive effort in
Palythoa caribaeorum. In Proceedings of
the 9th International Coral Reef Symposium, 2000. Bali. Bali: ICRS. p.
295.), and liberates a potent non-protein toxin called palytoxin (Seemann et al., 2009SEEMANN, P., Gernert, C., Schmitt, S., Mebs, D. and Hentschel, U.,
2009. Detection of hemolytic bacteria from Palythoa caribaeorum
(Cnidaria, Zoantharia) using a novel palytoxin-screening assay. Antonie
Leeuwenhoek International Journal of Micobiology, vol. 96, no. 4, p. 405-411.
http://dx.doi.org/10.1007/s10482-009-9353-4.
http://dx.doi.org/10.1007/s10482-009-935...
).This cnidarian is
popularly known as “baba-de-boi” (“cattle spittle”) as it secretes a very viscous
mucus over the surface of the colony during low tides that can shelter other marine
microorganisms (Ainsworth et al., 2010Ainsworth, TD., Thurber, RV. and Gates, RD., 2010. The future of
coral reefs: a microbial perspective. Trends in Ecology & Evolution, vol.
25, no. 4, p. 233-240. http://dx.doi.org/10.1016/j.tree.2009.11.001.
PMid:20006405
http://dx.doi.org/10.1016/j.tree.2009.11...
).
The bacterium Alcanivorax dieselolei was originally isolated from
seawater samples and marine sediments contaminated by oil in the Pacific Ocean and
described by Liu and Shao (2005)Liu, C. and Shao, Z., 2005. Alcanivorax dieselolei sp. nov., a novel
alkane-degrading bacterium isolated from sea water and deep-sea sediment.
International Journal of Systematic and Evolutionary Microbiology, vol. 55, no.
Pt 3, p. 1181-1186. http://dx.doi.org/10.1099/ijs.0.63443-0.
PMid:15879252
http://dx.doi.org/10.1099/ijs.0.63443-0...
. These
bacteria, as well as other species of the same genus, predominate in temperate
marine environments impacted by oil (Cappello et
al., 2007Cappello, S., Denaro, R., Genovese, M., Giuliano, L. and Yakimov,
MM., 2007. Predominant growth of Alcanivorax during experiments
on “oil spill bioremediation” in mesocosms. Microbiological Research, vol. 162,
no. 2, p. 185-190. http://dx.doi.org/10.1016/j.micres.2006.05.010.
PMid:16831537
http://dx.doi.org/10.1016/j.micres.2006....
).
The present work represents the first record of A. dieselolei, a specie capable of degrading petroleum derivatives, in the Atlantic Ocean and associated with the zoanthid P. caribaeorum. The potential biotechnological applications of the bacterium are discussed.
2 Material and Methods
Mucus was collected from exposed reef colonies of P. caribaeorum (Figure 1) during low tide periods in May/2010 at Porto de Galinhas on the southern coast of Pernambuco State, Brazil (8°30’24’’S; 34°59’52’’W) (Figure 2). Mucus samples were scraped and placed in sterile 50 mL centrifuge tubes. Heterotrophic bacteria were isolated from the mucus by inoculating 2 mL of the collected mucus onto Marine Agar 2216 (Difco®) using the pour plate technique and incubating the cultures at 30 °C for five days.
DNA was extracted from individual bacterial colonies using the thermal shock technique, in which a small quantity of material from each colony was collected and resuspended in 100 µl of ultrapure sterilized water, exposed to temperatures of 98 °C for 10 min. and –20 °C for 10 min. and subsequently centrifuged; the supernatants were transferred to sterile tubes.
The total DNA from each sample was used as a template for amplifying the 16S rDNA
segments of Eubacteria using the primers 27F (5’ AGA GTT TGA TCM TGG CTC AG 3’) and
1492R (5’ TAC GCY TAC CTT GTT ACG ACT T 3’) (Rohwer
et al., 2002Rohwer, F., Seguritan, V., Azam, F. and Knowlton, N., 2002.
Diversity and distribution of coral associated bacteria. Marine Ecology Progress
Series, vol. 243, p. 1-10.
http://dx.doi.org/10.3354/meps243001.
http://dx.doi.org/10.3354/meps243001...
). The PCR reactions were performed in a final volume of 50
µl that included: 50 ng of the template DNA, 10 pmol of each primer, 200 µM dNTPs, 5
µl PCR buffer, 5 U of Taq polymerase for DNA (Fermentas®), and 22 µl of
ultrapure sterilized water. The thermocycler program consisted of: (1) 5 minutes at
94 °C; (2) 30 cycles of 1 minute at 94 °C, 1 minute at 62 °C, and 3 minutes at 72
°C, and; (3) 10 minutes at 72 °C. The PCR products were purified using the QIAquick
PCR Purification kit (QIAGEN®).
The sequencing reactions were performed using BigDye® (ABI) and DYEnamic ET (Amersham Biosciences®) chemistry for the ABI Prism (Model 3100). The reactions were performed in a final volume of 10 µl containing: 100 ng of template DNA (the amplification products), 2 µl of marked nucleotides, 25X PCR buffer (20 mM Tris-HCl pH=8.4), 25 pmoles of each primer (forward and reverse), and previously sterilized Milli-Q water (q.s.p. 20 μl). These reactions were performed using the same program described above for the PCR reactions.
The qualities of the resulting DNA sequences were evaluated using Sequencing Analysis
3.4 software (Applied Biosystems®, Foster City, CA, EUA). The sequences
were submitted to nucleotide similarity consultations of the data available at
GenBank, on the the NCBI (National Center for Biotechnology Information) web site
using the BLASTn algorithm (Altschul et al., 1990Altschul, SF., Gish, W., Miller, W., Myers, EW. and Lipman, DJ.,
1990. Basic local alignment search tool. Journal of Molecular Biology, vol. 215,
no. 3, p. 403-410. http://dx.doi.org/10.1016/S0022-2836(05)80360-2.
PMid:2231712
http://dx.doi.org/10.1016/S0022-2836(05)...
).
3 Results and Discussion
Fifty bacterial isolates were obtained from the mucus of P.
caribaeorum collected on reefs in Porto de Galinhas, Pernambuco State,
Brazil. The dominant group among the isolated bacteria was γ-Proteobacteria, with 36
isolates (72%), followed by α-Proteobacteria and Actinobacteria with six isolates
each (12%), and Firmicutes with two isolates (4%); one isolate of A.
dieselolei was encountered among the γ-Proteobacteria. Comparisons of
the nucleotide sequence of the 16S rDNA gene of the bacterium isolated from
P. caribaeorum mucus with sequences deposited in the GenBank
indicated similarities 100% with the sequences of the group that performed the
complete genome sequence of A. dieselolei Type Strain B5 (Lai et al., 2012Lai, Q., Li, W. and Shao, Z., 2012. Complete genome sequence of type
strain B5. Alcanivorax dieseloleiJournal of Bacteriology, vol.
194, no. 23, p. 6674. http://dx.doi.org/10.1128/JB.01813-12.
PMid:23144414
http://dx.doi.org/10.1128/JB.01813-12...
). The nucleotide sequence was
deposited in GenBank with the accession number KF545933.
We therefore report here the first known association of the bacteria A.
dieselolei with the zoanthid P. caribaeorum. Sequences
of this bacteria had only previously been obtained from seawater and marine sediment
samples (Liu and Shao, 2005Liu, C. and Shao, Z., 2005. Alcanivorax dieselolei sp. nov., a novel
alkane-degrading bacterium isolated from sea water and deep-sea sediment.
International Journal of Systematic and Evolutionary Microbiology, vol. 55, no.
Pt 3, p. 1181-1186. http://dx.doi.org/10.1099/ijs.0.63443-0.
PMid:15879252
http://dx.doi.org/10.1099/ijs.0.63443-0...
; Tapilatu et al., 2010Tapilatu, Y., Acquaviva, M., Guigue, C., Miralles, G., Bertrand, JC.
and Cuny, P., 2010. Isolation of alkane-degrading bacteria from deep-sea
Mediterranean sediments. Letters in Applied Microbiology, vol. 50, no. 2, p.
234-236. http://dx.doi.org/10.1111/j.1472-765X.2009.02766.x.
PMid:19943883
http://dx.doi.org/10.1111/j.1472-765X.20...
), as is true for most of
the other isolates and 16S rRNA gene sequences of other bacterial strains of the
genus Alcanivorax (Liu and Shao,
2005Liu, C. and Shao, Z., 2005. Alcanivorax dieselolei sp. nov., a novel
alkane-degrading bacterium isolated from sea water and deep-sea sediment.
International Journal of Systematic and Evolutionary Microbiology, vol. 55, no.
Pt 3, p. 1181-1186. http://dx.doi.org/10.1099/ijs.0.63443-0.
PMid:15879252
http://dx.doi.org/10.1099/ijs.0.63443-0...
; Cappello et al., 2007Cappello, S., Denaro, R., Genovese, M., Giuliano, L. and Yakimov,
MM., 2007. Predominant growth of Alcanivorax during experiments
on “oil spill bioremediation” in mesocosms. Microbiological Research, vol. 162,
no. 2, p. 185-190. http://dx.doi.org/10.1016/j.micres.2006.05.010.
PMid:16831537
http://dx.doi.org/10.1016/j.micres.2006....
;
Rivas et al., 2007Rivas, R., García-Fraile, P., Peix, A., Mateos, PF.,
Martínez-Molina, E. and Velázquez, E., 2007. Alcanivorax balearicus sp. nov.,
isolated from Lake Martel. International Journal of Systematic and Evolutionary
Microbiology, vol. 57, no. 6, p. 1331-1335.
http://dx.doi.org/10.1099/ijs.0.64912-0. PMid:17551053
http://dx.doi.org/10.1099/ijs.0.64912-0...
; Wu et al., 2009Wu, Y., Lai, Q., Zhou, Z., Qiao, N., Liu, C. and Shao, Z., 2009.
Alcanivorax hongdengensis sp. nov., an alkane-degrading bacterium isolated from
surface seawater of the straits of Malacca and Singapore, producing a
lipopeptide as its biosurfactant. International Journal of Systematic and
Evolutionary Microbiology, vol. 59, no. 6, p. 1474-1479.
http://dx.doi.org/10.1099/ijs.0.001552-0. PMid:19502338
http://dx.doi.org/10.1099/ijs.0.001552-0...
). Some presently
non-identified strains of the genus Alcanivorax have been isolated
from marine invertebrates such as sponges and gorgonians (Sfanos et al., 2005Sfanos, K., Harmody, D., Dang, P., Ledger, A., Pomponi, S.,
McCarthy, P. and Lopez, J., 2005. A molecular systematic survey of cultured
microbial associates of deep-water marine invertebrates. Systematic and Applied
Microbiology, vol. 28, no. 3, p. 242-264.
http://dx.doi.org/10.1016/j.syapm.2004.12.002. PMid:15900971
http://dx.doi.org/10.1016/j.syapm.2004.1...
), but our current report is the first
record of any association between A. dieselolei and P.
caribaeorum, and its first reported occurrence in the Atlantic
Ocean.
Among the hydrocarbonoclastic bacteria, the genus Alcanivorax
comprises Gram-negative, aerobic, and halophytic species that have the capacity to
metabolize alkanes hydrocarbons as a carbon source and for energy and have been used
in bioremediation projects in polluted marine environments (Yakimov et al., 1998YAKIMOV, MM., GOLYSHIN, PN., LANG, S., MOORE, ERB., ABRAHAM, WR.,
LÜNSDORF, H. and TIMMIS, KN., 1998. Alcanivorax borkumensis gen. nov., sp. nov.,
a new hydrocarbon-degrading and surfactant-producing marine bacterium.
International Journal of Systematic Bacteriology, vol. 48, no. 2, p. 339-348.
http://dx.doi.org/10.1099/00207713-48-2-339.
http://dx.doi.org/10.1099/00207713-48-2-...
; Liu and
Shao, 2005Liu, C. and Shao, Z., 2005. Alcanivorax dieselolei sp. nov., a novel
alkane-degrading bacterium isolated from sea water and deep-sea sediment.
International Journal of Systematic and Evolutionary Microbiology, vol. 55, no.
Pt 3, p. 1181-1186. http://dx.doi.org/10.1099/ijs.0.63443-0.
PMid:15879252
http://dx.doi.org/10.1099/ijs.0.63443-0...
). The genus Alcanivorax comprises six
described species: Alcanivorax borkumensis (Yakimov et al., 1998YAKIMOV, MM., GOLYSHIN, PN., LANG, S., MOORE, ERB., ABRAHAM, WR.,
LÜNSDORF, H. and TIMMIS, KN., 1998. Alcanivorax borkumensis gen. nov., sp. nov.,
a new hydrocarbon-degrading and surfactant-producing marine bacterium.
International Journal of Systematic Bacteriology, vol. 48, no. 2, p. 339-348.
http://dx.doi.org/10.1099/00207713-48-2-339.
http://dx.doi.org/10.1099/00207713-48-2-...
), Alcanivorax venustensis,
Alcanivorax jadensis (Fernández-Martínez, 2003), Alcanivorax
dieselolei (Liu and Shao,
2005Liu, C. and Shao, Z., 2005. Alcanivorax dieselolei sp. nov., a novel
alkane-degrading bacterium isolated from sea water and deep-sea sediment.
International Journal of Systematic and Evolutionary Microbiology, vol. 55, no.
Pt 3, p. 1181-1186. http://dx.doi.org/10.1099/ijs.0.63443-0.
PMid:15879252
http://dx.doi.org/10.1099/ijs.0.63443-0...
), Alcanivorax balearicus (Rivas et al., 2007Rivas, R., García-Fraile, P., Peix, A., Mateos, PF.,
Martínez-Molina, E. and Velázquez, E., 2007. Alcanivorax balearicus sp. nov.,
isolated from Lake Martel. International Journal of Systematic and Evolutionary
Microbiology, vol. 57, no. 6, p. 1331-1335.
http://dx.doi.org/10.1099/ijs.0.64912-0. PMid:17551053
http://dx.doi.org/10.1099/ijs.0.64912-0...
), and Alcanivorax
hongdengensis (Wu et al.,
2009Wu, Y., Lai, Q., Zhou, Z., Qiao, N., Liu, C. and Shao, Z., 2009.
Alcanivorax hongdengensis sp. nov., an alkane-degrading bacterium isolated from
surface seawater of the straits of Malacca and Singapore, producing a
lipopeptide as its biosurfactant. International Journal of Systematic and
Evolutionary Microbiology, vol. 59, no. 6, p. 1474-1479.
http://dx.doi.org/10.1099/ijs.0.001552-0. PMid:19502338
http://dx.doi.org/10.1099/ijs.0.001552-0...
).
The fact that A. dieselolei was isolated from colonies of P.
caribaeorum is quite intriguing because, while the beach at Porto de
Galinhas has been heavily visited by tourists, it does not appear to be contaminated
to any significant degree by petroleum compounds in spite of its proximity to the
shipping port at Suape. On the other hand, large numbers of swimmers and divers do
visit this beach and use solar protection lotions containing hydrocarbons such as
benzophenone, oxybenzone, or parabens. These compounds may serve as energy and
carbon sources for bacteria that metabolize only alkanes and reject other carbon
resources such as sugars and amino acids (Lorenzo,
2006Lorenzo, V., 2006. Blueprint of an oil-eating bacterium. Nature
Biotechnology, vol. 24, no. 8, p. 952-953.
http://dx.doi.org/10.1038/nbt0806-952. PMid:16900140
http://dx.doi.org/10.1038/nbt0806-952...
).
The presence of the bacteria A. dieselolei in P.
caribaeorum mucus might also be at least partially explained by the
presence of high concentrations of phosphorus and nitrogen compounds (commonly used
in bioremediation projects to break bacterial dormancy) in that milieu (Cappello et al., 2007Cappello, S., Denaro, R., Genovese, M., Giuliano, L. and Yakimov,
MM., 2007. Predominant growth of Alcanivorax during experiments
on “oil spill bioremediation” in mesocosms. Microbiological Research, vol. 162,
no. 2, p. 185-190. http://dx.doi.org/10.1016/j.micres.2006.05.010.
PMid:16831537
http://dx.doi.org/10.1016/j.micres.2006....
). It is also known that
bacteria of the genus Vibrio, which can fix nitrogen, are
associated with this same zoanthid (Chimetto et
al., 2008Chimetto, LA., Brocchi, M., Thompson, CC., Martins, RCR., Ramos, HR.
and Thompson, FL., 2008. Vibrios dominate as culturable nitrogen-fixing bacteria
of the Brazilian coral . Mussismilia hispidaSystematic and
Applied Microbiology, vol. 31, no. 4, p. 312-319.
http://dx.doi.org/10.1016/j.syapm.2008.06.001. PMid:18678453
http://dx.doi.org/10.1016/j.syapm.2008.0...
) so that A. dieselolei may be using these
compounds as energy sources as these bacteria show denitrification activity and
Alcanivorax strains are known to contribute to the degradation
of nitrogen and phosphorus compounds that accumulate in organically enriched coastal
areas (Nakano et al., 2009Nakano, M., Okunishi, S., Tanaka, R. and Maeda, H., 2009.
Denitrifying activity and homologous enzyme analysis of Alcanivorax dieselolei
strain N1203. Biocontrol Science, vol. 14, no. 3, p. 97-105.
http://dx.doi.org/10.4265/bio.14.97. PMid:19785282
http://dx.doi.org/10.4265/bio.14.97...
).
Another possible explanation for the presence of this bacterium in association with
P. caribaeorum would be the natural production of aliphatic
compounds by that animal. According to Yakimov et
al. (2007)Yakimov, MM., Timmis, KN. and Golyshin, PN., 2007. Obligate
oil-degrading marine bacteria. Current Opinion in Biotechnology, vol. 18, no. 3,
p. 257-266. http://dx.doi.org/10.1016/j.copbio.2007.04.006.
PMid:17493798
http://dx.doi.org/10.1016/j.copbio.2007....
, the isolation of the bacteria A. borkumensis
from marine invertebrates may reflect the existence of specific ecological niches
containing hydrocarbons produced by those animals themselves. The presence of
A. borkumensis in non-polluted environments (Kasai et al., 2001Kasai, Y., Kishira, H., Syutsubo, K. and Harayama, S., 2001.
Molecular detection of marine bacterial populations on beaches contaminated by
the . Nakhodka tanker oil-spill accidentEnvironmental
Microbiology, vol. 3, no. 4, p. 246-255.
http://dx.doi.org/10.1046/j.1462-2920.2001.00185.x.
PMid:11359510
http://dx.doi.org/10.1046/j.1462-2920.20...
) indicates that A.
dieselolei might likewise naturally exist in environments not polluted
by oil. It will be necessary to monitor the Porto de Galinhas beach area for
possible environmental impacts in any case, as these bacteria can be used as
bioindicators of water contamination by either high or low levels of long-chain
hydrocarbons (Fernández-Martínez et al.,
2003Fernández-Martínez, J., Pujalte, MJ., García-Martínez, J., Mata, M.,
Garay, E. and Rodríguez-Valeral, F., 2003. Description of Alcanivorax
venustensis sp. nov. and reclassification of Fundibacter jadensis DSM 1 21 78T
(Bruns and Berthe-Corti 1999) as comb. nov., members of the emended genus .
Alcanivorax
jadensisAlcanivoraxInternational Journal of Systematic
and Evolutionary Microbiology, vol. 53, no. 1, p. 331-338.
http://dx.doi.org/10.1099/ijs.0.01923-0. PMid:12656192
http://dx.doi.org/10.1099/ijs.0.01923-0...
).
Other genera (similarity indices ≥ 97%) with bacterial species with known capabilities to degrade alkanes derived from petroleum were also found associated with the secreted mucus of P. caribaeorum in the present work, including Altererytrobacter, Pseudomonas, Rhodococcus, and Stappia – which is interesting from the point of view of possible biotechnological applications. The present study also represents the first report of the association of these bacteria with the zoanthid P. caribaeorum, a cnidarian that has been only poorly examined in terms of its associated microbiota in spite of its abundance in most reef environments in the western Atlantic.
Acknowledgements
The authors would like to thank Fundação de Amparo à Ciência e Tecnologia do Estado de Pernambuco for their financial support to research and for the Master’s grant awarded to F.F.C., as well as Dra. Michelle Oliveira for her help in cultivating the bacteria in the laboratory.
References
- Acosta, A. and Asbahr, M., 2000. Reproductive effort in Palythoa caribaeorum. In Proceedings of the 9th International Coral Reef Symposium, 2000. Bali. Bali: ICRS. p. 295.
- Ainsworth, TD., Thurber, RV. and Gates, RD., 2010. The future of coral reefs: a microbial perspective. Trends in Ecology & Evolution, vol. 25, no. 4, p. 233-240. http://dx.doi.org/10.1016/j.tree.2009.11.001. PMid:20006405
» http://dx.doi.org/10.1016/j.tree.2009.11.001 - Altschul, SF., Gish, W., Miller, W., Myers, EW. and Lipman, DJ., 1990. Basic local alignment search tool. Journal of Molecular Biology, vol. 215, no. 3, p. 403-410. http://dx.doi.org/10.1016/S0022-2836(05)80360-2. PMid:2231712
» http://dx.doi.org/10.1016/S0022-2836(05)80360-2 - Cappello, S., Denaro, R., Genovese, M., Giuliano, L. and Yakimov, MM., 2007. Predominant growth of Alcanivorax during experiments on “oil spill bioremediation” in mesocosms. Microbiological Research, vol. 162, no. 2, p. 185-190. http://dx.doi.org/10.1016/j.micres.2006.05.010. PMid:16831537
» http://dx.doi.org/10.1016/j.micres.2006.05.010 - Castro, AP., Araújo JUNIOR, SD., Reis, AMM., Moura, RL., Francini-Filho, RB., Pappas JUNIOR, G., Rodrigues, TB., Thompson, FL. and Krüger, RH., 2010. Bacterial community associated with healthy and diseased reef coral Mussismilia hispida from eastern Brazil. Microbial Ecology, vol. 59, no. 4, p. 658-667. http://dx.doi.org/10.1007/s00248-010-9646-1. PMid:20352207
» http://dx.doi.org/10.1007/s00248-010-9646-1 - Chimetto, LA., Brocchi, M., Gondo, M., Thompson, CC., Gomez-Gil, B. and Thompson, FL., 2009. Genomic diversity of vibrios associated with the Brazilian coral Mussismilia hispida and its sympatric zoanthids (Palythoa caribaeorum, Palythoa variabilis and ). Zoanthus solanderiJournal of Applied Microbiology, vol. 106, no. 6, p. 1818-1826. http://dx.doi.org/10.1111/j.1365-2672.2009.04149.x. PMid:19291243
» http://dx.doi.org/10.1111/j.1365-2672.2009.04149.x - Chimetto, LA., Brocchi, M., Thompson, CC., Martins, RCR., Ramos, HR. and Thompson, FL., 2008. Vibrios dominate as culturable nitrogen-fixing bacteria of the Brazilian coral . Mussismilia hispidaSystematic and Applied Microbiology, vol. 31, no. 4, p. 312-319. http://dx.doi.org/10.1016/j.syapm.2008.06.001. PMid:18678453
» http://dx.doi.org/10.1016/j.syapm.2008.06.001 - Chimetto, LA., Cleenwerck, I., Alves JUNIOR, N., Silva, BS., Brocchi, M., Willems, A., De Vos, P. and Thompson, FL., 2011. Vibrio communis sp. nov., isolated from the marine animals Mussismilia hispida, , . Phyllogorgia dilatataPalythoa caribaeorum, Palythoa variabilis and Litopenaeus vannameiInternational Journal of Systematic and Evolutionary Microbiology, vol. 61, no. 2, p. 362-368. http://dx.doi.org/10.1099/ijs.0.019729-0. PMid:20305064
» http://dx.doi.org/10.1099/ijs.0.019729-0 - Fernández-Martínez, J., Pujalte, MJ., García-Martínez, J., Mata, M., Garay, E. and Rodríguez-Valeral, F., 2003. Description of Alcanivorax venustensis sp. nov. and reclassification of Fundibacter jadensis DSM 1 21 78T (Bruns and Berthe-Corti 1999) as comb. nov., members of the emended genus . Alcanivorax jadensisAlcanivoraxInternational Journal of Systematic and Evolutionary Microbiology, vol. 53, no. 1, p. 331-338. http://dx.doi.org/10.1099/ijs.0.01923-0. PMid:12656192
» http://dx.doi.org/10.1099/ijs.0.01923-0 - Kasai, Y., Kishira, H., Syutsubo, K. and Harayama, S., 2001. Molecular detection of marine bacterial populations on beaches contaminated by the . Nakhodka tanker oil-spill accidentEnvironmental Microbiology, vol. 3, no. 4, p. 246-255. http://dx.doi.org/10.1046/j.1462-2920.2001.00185.x. PMid:11359510
» http://dx.doi.org/10.1046/j.1462-2920.2001.00185.x - Lai, Q., Li, W. and Shao, Z., 2012. Complete genome sequence of type strain B5. Alcanivorax dieseloleiJournal of Bacteriology, vol. 194, no. 23, p. 6674. http://dx.doi.org/10.1128/JB.01813-12. PMid:23144414
» http://dx.doi.org/10.1128/JB.01813-12 - Liu, C. and Shao, Z., 2005. Alcanivorax dieselolei sp. nov., a novel alkane-degrading bacterium isolated from sea water and deep-sea sediment. International Journal of Systematic and Evolutionary Microbiology, vol. 55, no. Pt 3, p. 1181-1186. http://dx.doi.org/10.1099/ijs.0.63443-0. PMid:15879252
» http://dx.doi.org/10.1099/ijs.0.63443-0 - Lorenzo, V., 2006. Blueprint of an oil-eating bacterium. Nature Biotechnology, vol. 24, no. 8, p. 952-953. http://dx.doi.org/10.1038/nbt0806-952. PMid:16900140
» http://dx.doi.org/10.1038/nbt0806-952 - Meikle, P., RICHARDS, GN. and YELLOWLEES, D., 1988. Structural investigations on the mucus from six species of coral. Marine Biology, vol. 99, no. 2, p. 187-193. http://dx.doi.org/10.1007/BF00391980.
» http://dx.doi.org/10.1007/BF00391980 - Menezes, CBA., Bonugli-Santos, RC., Miqueletto, PB., Passarini, MRZ., Silva, CHD., Justo, MR., Leal, RR., Fantinatti-Garboggini, F., Oliveira, VM., Berlinck, RGS. and Sette, LD., 2010. Microbial diversity associated with algae, ascidians and sponges from the north coast of São Paulo state, Brazil. Microbiological Research, vol. 165, no. 6, p. 466-482. http://dx.doi.org/10.1016/j.micres.2009.09.005. PMid:19879115
» http://dx.doi.org/10.1016/j.micres.2009.09.005 - MUELLER, E. and HAYWICK, DW., 1995. Sediment assimilation and calcification by the Western Atlantic reef zoanthid, Palythoa caribaeorum.Bulletin de l'Institut Oceanographique, vol. 14, p. 89-100.
- Nakano, M., Okunishi, S., Tanaka, R. and Maeda, H., 2009. Denitrifying activity and homologous enzyme analysis of Alcanivorax dieselolei strain N1203. Biocontrol Science, vol. 14, no. 3, p. 97-105. http://dx.doi.org/10.4265/bio.14.97. PMid:19785282
» http://dx.doi.org/10.4265/bio.14.97 - PÉREZ, CD., VILA-NOVA, DA. and SANTOS, AM., 2005. Associated community with the zoanthid (Duchassaing & Michelotti, 1860) (Cnidaria, Anthozoa) from littoral of Pernambuco, Brazil. Palythoa caribaeorumHydrobiologia, vol. 548, no. 1, p. 207-215. http://dx.doi.org/10.1007/s10750-005-5441-2.
» http://dx.doi.org/10.1007/s10750-005-5441-2 - Rivas, R., García-Fraile, P., Peix, A., Mateos, PF., Martínez-Molina, E. and Velázquez, E., 2007. Alcanivorax balearicus sp. nov., isolated from Lake Martel. International Journal of Systematic and Evolutionary Microbiology, vol. 57, no. 6, p. 1331-1335. http://dx.doi.org/10.1099/ijs.0.64912-0. PMid:17551053
» http://dx.doi.org/10.1099/ijs.0.64912-0 - Rohwer, F., Seguritan, V., Azam, F. and Knowlton, N., 2002. Diversity and distribution of coral associated bacteria. Marine Ecology Progress Series, vol. 243, p. 1-10. http://dx.doi.org/10.3354/meps243001.
» http://dx.doi.org/10.3354/meps243001 - SEBENS, KP, 1982. Intertidal distribution of zoanthids on the Caribbean coast of Panama: effects of predation and dessication. Bulletin of Marine Science, vol. 32, p. 316-335.
- SEEMANN, P., Gernert, C., Schmitt, S., Mebs, D. and Hentschel, U., 2009. Detection of hemolytic bacteria from Palythoa caribaeorum (Cnidaria, Zoantharia) using a novel palytoxin-screening assay. Antonie Leeuwenhoek International Journal of Micobiology, vol. 96, no. 4, p. 405-411. http://dx.doi.org/10.1007/s10482-009-9353-4.
» http://dx.doi.org/10.1007/s10482-009-9353-4 - Sfanos, K., Harmody, D., Dang, P., Ledger, A., Pomponi, S., McCarthy, P. and Lopez, J., 2005. A molecular systematic survey of cultured microbial associates of deep-water marine invertebrates. Systematic and Applied Microbiology, vol. 28, no. 3, p. 242-264. http://dx.doi.org/10.1016/j.syapm.2004.12.002. PMid:15900971
» http://dx.doi.org/10.1016/j.syapm.2004.12.002 - Tapilatu, Y., Acquaviva, M., Guigue, C., Miralles, G., Bertrand, JC. and Cuny, P., 2010. Isolation of alkane-degrading bacteria from deep-sea Mediterranean sediments. Letters in Applied Microbiology, vol. 50, no. 2, p. 234-236. http://dx.doi.org/10.1111/j.1472-765X.2009.02766.x. PMid:19943883
» http://dx.doi.org/10.1111/j.1472-765X.2009.02766.x - Trindade-Silva, AE., Rua, C., Silva, GGZ., Dutilh, BE., Moreira, APB., Edwards, RA., Hajdu, E., Lobo-Hajdu, G., Vasconcelos, AT., Berlinck, RGS. and Thompson, FL., 2012. Taxonomic and functional microbial signatures of the endemic marine sponge Arenosclera brasiliensis. PLoS ONE, vol. 7, no. 7, p. e39905. http://dx.doi.org/10.1371/journal.pone.0039905. PMid:22768320
» http://dx.doi.org/10.1371/journal.pone.0039905 - Wegley, L., Edwards, R., Rodriguez-Brito, B., Liu, H. and Rohwer, F., 2007. Metagenomic analysis of the microbial community associated with the coral Porites astreoides. Environmental Microbiology, vol. 9, no. 11, p. 2707-2719. http://dx.doi.org/10.1111/j.1462-2920.2007.01383.x. PMid:17922755
» http://dx.doi.org/10.1111/j.1462-2920.2007.01383.x - Wu, Y., Lai, Q., Zhou, Z., Qiao, N., Liu, C. and Shao, Z., 2009. Alcanivorax hongdengensis sp. nov., an alkane-degrading bacterium isolated from surface seawater of the straits of Malacca and Singapore, producing a lipopeptide as its biosurfactant. International Journal of Systematic and Evolutionary Microbiology, vol. 59, no. 6, p. 1474-1479. http://dx.doi.org/10.1099/ijs.0.001552-0. PMid:19502338
» http://dx.doi.org/10.1099/ijs.0.001552-0 - YAKIMOV, MM., GOLYSHIN, PN., LANG, S., MOORE, ERB., ABRAHAM, WR., LÜNSDORF, H. and TIMMIS, KN., 1998. Alcanivorax borkumensis gen. nov., sp. nov., a new hydrocarbon-degrading and surfactant-producing marine bacterium. International Journal of Systematic Bacteriology, vol. 48, no. 2, p. 339-348. http://dx.doi.org/10.1099/00207713-48-2-339.
» http://dx.doi.org/10.1099/00207713-48-2-339 - Yakimov, MM., Timmis, KN. and Golyshin, PN., 2007. Obligate oil-degrading marine bacteria. Current Opinion in Biotechnology, vol. 18, no. 3, p. 257-266. http://dx.doi.org/10.1016/j.copbio.2007.04.006. PMid:17493798
» http://dx.doi.org/10.1016/j.copbio.2007.04.006
-
(With 2 Figures)
Publication Dates
-
Publication in this collection
May 2015
History
-
Received
13 Sept 2013 -
Accepted
20 Jan 2014