MALDI-TOF MS for the identification of bacteria from Acropora cervicornis samples with White Band Disease
White Band Disease (WBD) is one of the most harmful coral syndromes in the Caribbean, deteriorating the structure of many corals during the eighties and nineties. Since its etiology is not entirely known, the use of techniques aimed a t identifying the associated agents could provide an approach to...
- Autores:
-
Fennix Agudelo, Mary Andrea
Zarza González, Esteban
Sánchez Quitian, Zilpa Adriana
Castellanos, Nuri
Martínez Santos, Juan Carlos
Parra Giraldo, Claudia Marcela
Chavarro Mesa, Edisson
- Tipo de recurso:
- Fecha de publicación:
- 2022
- Institución:
- Universidad Tecnológica de Bolívar
- Repositorio:
- Repositorio Institucional UTB
- Idioma:
- eng
- OAI Identifier:
- oai:repositorio.utb.edu.co:20.500.12585/12380
- Acceso en línea:
- https://hdl.handle.net/20.500.12585/12380
- Palabra clave:
- Anthozoa;
Acropora;
Microorganisms
LEMB
- Rights
- openAccess
- License
- http://creativecommons.org/licenses/by-nc-nd/4.0/
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MALDI-TOF MS for the identification of bacteria from Acropora cervicornis samples with White Band Disease |
| title |
MALDI-TOF MS for the identification of bacteria from Acropora cervicornis samples with White Band Disease |
| spellingShingle |
MALDI-TOF MS for the identification of bacteria from Acropora cervicornis samples with White Band Disease Anthozoa; Acropora; Microorganisms LEMB |
| title_short |
MALDI-TOF MS for the identification of bacteria from Acropora cervicornis samples with White Band Disease |
| title_full |
MALDI-TOF MS for the identification of bacteria from Acropora cervicornis samples with White Band Disease |
| title_fullStr |
MALDI-TOF MS for the identification of bacteria from Acropora cervicornis samples with White Band Disease |
| title_full_unstemmed |
MALDI-TOF MS for the identification of bacteria from Acropora cervicornis samples with White Band Disease |
| title_sort |
MALDI-TOF MS for the identification of bacteria from Acropora cervicornis samples with White Band Disease |
| dc.creator.fl_str_mv |
Fennix Agudelo, Mary Andrea Zarza González, Esteban Sánchez Quitian, Zilpa Adriana Castellanos, Nuri Martínez Santos, Juan Carlos Parra Giraldo, Claudia Marcela Chavarro Mesa, Edisson |
| dc.contributor.author.none.fl_str_mv |
Fennix Agudelo, Mary Andrea Zarza González, Esteban Sánchez Quitian, Zilpa Adriana Castellanos, Nuri Martínez Santos, Juan Carlos Parra Giraldo, Claudia Marcela Chavarro Mesa, Edisson |
| dc.subject.keywords.spa.fl_str_mv |
Anthozoa; Acropora; Microorganisms |
| topic |
Anthozoa; Acropora; Microorganisms LEMB |
| dc.subject.armarc.none.fl_str_mv |
LEMB |
| description |
White Band Disease (WBD) is one of the most harmful coral syndromes in the Caribbean, deteriorating the structure of many corals during the eighties and nineties. Since its etiology is not entirely known, the use of techniques aimed a t identifying the associated agents could provide an approach to the composition of the populations involved in the deterioration of A. cervicornis. Therefore, this research's objective was to isolate and characterize bacteria associated with WBD in A. cervicornis from hanging nurseries of Rosario and San Bernardo Corals National Natural Park, Colombia, using MALDI-TO F MS and PCR. The results showed the presence of Bacillus cereus bacteria in individuals with bleaching signs from nurseries and transplant areas on Isla Tesoro and Isla Ceiner. One of these strains sequenced and phylogenetically analyzed was closely related t o Bacillus thuringiensis, with similarities of up to 99%. The presence of B. cereus could indicate a special relationship between this microorganism and WBD. However, it is imperative to carry out more studies on identifying the bacteria associated with this disease to support these findings. © 2022 Latin American and Caribbean Consortium of Engineering Institutions. All rights reserved. |
| publishDate |
2022 |
| dc.date.issued.none.fl_str_mv |
2022 |
| dc.date.accessioned.none.fl_str_mv |
2023-07-21T20:48:29Z |
| dc.date.available.none.fl_str_mv |
2023-07-21T20:48:29Z |
| dc.date.submitted.none.fl_str_mv |
2023 |
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info:eu-repo/semantics/article |
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info:eu-repo/semantics/draft |
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http://purl.org/coar/resource_type/c_6501 |
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draft |
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Fennix-Agudelo, M. A., Zarza-González, E., Sánchez-Quitian, Z. A., Merchán-Castellanos, N., Martinez-Santos, J. C., Parra-Giraldo, C. M., & Chavarro-Mesa, E. MALDI-TOF MS for the identification of bacteria from Acropora cervicornis samples with White Band. |
| dc.identifier.uri.none.fl_str_mv |
https://hdl.handle.net/20.500.12585/12380 |
| dc.identifier.doi.none.fl_str_mv |
10.18687/LACCEI2022.1.1.714 |
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Universidad Tecnológica de Bolívar |
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Repositorio Universidad Tecnológica de Bolívar |
| identifier_str_mv |
Fennix-Agudelo, M. A., Zarza-González, E., Sánchez-Quitian, Z. A., Merchán-Castellanos, N., Martinez-Santos, J. C., Parra-Giraldo, C. M., & Chavarro-Mesa, E. MALDI-TOF MS for the identification of bacteria from Acropora cervicornis samples with White Band. 10.18687/LACCEI2022.1.1.714 Universidad Tecnológica de Bolívar Repositorio Universidad Tecnológica de Bolívar |
| url |
https://hdl.handle.net/20.500.12585/12380 |
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eng |
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eng |
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info:eu-repo/semantics/openAccess |
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Attribution-NonCommercial-NoDerivatives 4.0 Internacional |
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openAccess |
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7 páginas |
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Cartagena de Indias |
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Proceedings of the LACCEI international Multi-conference for Engineering, Education and Technology |
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Fennix Agudelo, Mary Andrea1588a0ca-009d-4aca-8575-f9ce95f54615Zarza González, Esteban3e1d7a4b-51f9-4603-b44a-850b1d95d1e9Sánchez Quitian, Zilpa Adrianac6b988d9-38fb-4e79-9979-0a949652f1a0Castellanos, Nuric039f767-ebbe-4c94-97f8-c708ea1ff549Martínez Santos, Juan Carlos480cc438-b02b-45d2-bfc9-7bfa96f0271bParra Giraldo, Claudia Marcela70133c96-c400-49f2-9a70-df59fee3c0c7Chavarro Mesa, Edisson800ba77d-129a-48f9-bc50-2a8c25be12082023-07-21T20:48:29Z2023-07-21T20:48:29Z20222023Fennix-Agudelo, M. A., Zarza-González, E., Sánchez-Quitian, Z. A., Merchán-Castellanos, N., Martinez-Santos, J. C., Parra-Giraldo, C. M., & Chavarro-Mesa, E. MALDI-TOF MS for the identification of bacteria from Acropora cervicornis samples with White Band.https://hdl.handle.net/20.500.12585/1238010.18687/LACCEI2022.1.1.714Universidad Tecnológica de BolívarRepositorio Universidad Tecnológica de BolívarWhite Band Disease (WBD) is one of the most harmful coral syndromes in the Caribbean, deteriorating the structure of many corals during the eighties and nineties. Since its etiology is not entirely known, the use of techniques aimed a t identifying the associated agents could provide an approach to the composition of the populations involved in the deterioration of A. cervicornis. Therefore, this research's objective was to isolate and characterize bacteria associated with WBD in A. cervicornis from hanging nurseries of Rosario and San Bernardo Corals National Natural Park, Colombia, using MALDI-TO F MS and PCR. The results showed the presence of Bacillus cereus bacteria in individuals with bleaching signs from nurseries and transplant areas on Isla Tesoro and Isla Ceiner. One of these strains sequenced and phylogenetically analyzed was closely related t o Bacillus thuringiensis, with similarities of up to 99%. The presence of B. cereus could indicate a special relationship between this microorganism and WBD. However, it is imperative to carry out more studies on identifying the bacteria associated with this disease to support these findings. © 2022 Latin American and Caribbean Consortium of Engineering Institutions. All rights reserved.7 páginasapplication/pdfenghttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessAttribution-NonCommercial-NoDerivatives 4.0 Internacionalhttp://purl.org/coar/access_right/c_abf2Proceedings of the LACCEI international Multi-conference for Engineering, Education and TechnologyMALDI-TOF MS for the identification of bacteria from Acropora cervicornis samples with White Band Diseaseinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/drafthttp://purl.org/coar/resource_type/c_6501http://purl.org/coar/version/c_b1a7d7d4d402bccehttp://purl.org/coar/resource_type/c_2df8fbb1Anthozoa;Acropora;MicroorganismsLEMBCartagena de IndiasShinzato, C., Mungpakdee, S., Satoh, N., Shoguchi, E. A genomic approach to coral-dinoflagellate symbiosis: Studies of Acropora digitifera and Symbiodinium minutum (2014) Frontiers in Microbiology, 5 (JULY), art. no. 336. Cited 32 times. http://journal.frontiersin.org/Journal/10.3389/fmicb.2014.00336/full doi: 10.3389/fmicb.2014.00336Randall, C.J., Van Woesik, R. Some coral diseases track climate oscillations in the Caribbean (2017) Scientific Reports, 7 (1), art. no. 5719. Cited 22 times. www.nature.com/srep/index.html doi: 10.1038/s41598-017-05763-6Alvarez-Filip, L., Dulvy, N.K., Gill, J.A., Côté, I.M., Watkinson, A.R. Flattening of Caribbean coral reefs: Region-wide declines in architectural complexity (2009) Proceedings of the Royal Society B: Biological Sciences, 276 (1669), pp. 3019-3025. Cited 606 times. http://rspb.royalsocietypublishing.org/content/276/1669/3019.full.pdf doi: 10.1098/rspb.2009.0339Sweet, M.J., Séré, M.G. Ciliate communities consistently associated with coral diseases (2016) Journal of Sea Research, 113, pp. 119-131. Cited 38 times. www.elsevier.com/inca/publications/store/6/0/0/3/1/8 doi: 10.1016/j.seares.2015.06.008Popović, N.T., Kazazić, S.P., Strunjak-Perović, I., Čož-Rakovac, R. Differentiation of environmental aquatic bacterial isolates by MALDI-TOF MS (Open Access) (2017) Environmental Research, 152, pp. 7-16. Cited 44 times. http://www.elsevier.com/inca/publications/store/6/2/2/8/2/1/index.htt doi: 10.1016/j.envres.2016.09.020Singhal, N., Kumar, M., Kanaujia, P.K., Virdi, J.S. MALDI-TOF mass spectrometry: An emerging technology for microbial identification and diagnosis (2015) Frontiers in Microbiology, 6 (AUG), art. no. 791. Cited 848 times. http://journal.frontiersin.org/article/10.3389/fmicb.2015.00791/full doi: 10.3389/fmicb.2015.00791Vidal, L.M.R., Venas, T.M., Gonçalves, A.R.P., Mattsson, H.K., Silva, R.V.P., Nóbrega, M.S., Azevedo, G.P.R., (...), Thompson, C.C. Rapid screening of marine bacterial symbionts using MALDI-TOF MS (2020) Archives of Microbiology, 202 (8), pp. 2329-2336. Cited 4 times. link.springer.de/link/service/journals/00203/index.htm doi: 10.1007/s00203-020-01917-9Brauge, T., Trigueros, S., Briet, A., Debuiche, S., Leleu, G., Gassilloud, B., Wilhelm, A., (...), Midelet, G. MALDI-TOF Mass Spectrometry Fingerprinting Performance Versus 16S rDNA Sequencing to Identify Bacterial Microflora From Seafood Products and Sea Water Samples (Open Access) (2021) Frontiers in Marine Science, 8, art. no. 650116. Cited 4 times. https://www.frontiersin.org/journals/marine-science# doi: 10.3389/fmars.2021.650116Daniels, C.A., Baumgarten, S., Yum, L.K., Michell, C.T., Bayer, T., Arif, C., Roder, C., (...), Voolstra, C.R. Metatranscriptome analysis of the reef-building coral Orbicella faveolata indicates holobiont response to coral disease (Open Access) (2015) Frontiers in Marine Science, 2 (SEP), art. no. 62. Cited 54 times. http://journal.frontiersin.org/article/10.3389/fmars.2015.00062/full doi: 10.3389/fmars.2015.00062Garcia, G.D., Gregoracci, G.B., Santos, E.O., Meirelles, P.M., Silva, G.G.Z., Edwards, R., Sawabe, T., (...), Thompson, F.L. Metagenomic Analysis of Healthy and White Plague-Affected Mussismilia braziliensis Corals (Open Access) (2013) Microbial Ecology, 65 (4), pp. 1076-1086. Cited 66 times. link.springer.de/link/service/journals/00248/index.htm doi: 10.1007/s00248-012-0161-4Kalimutho, M., Ahmad, A., Kassim, Z. Isolation, characterization and identification of bacteria associated with mucus of Acropora cervicornis coral from Bidong Island, Terengganu, Malaysia (Open Access) (2007) Malaysian Journal of Science, 26 (2), pp. 27-39. Cited 12 times.Rodriguez-Temporal, D., Perez-Risco, D., Struzka, E.A., Mas, M., Alcaide, F. Evaluation of two protein extraction protocols based on freezing and mechanical disruption for identifying nontuberculous mycobacteria by matrix-assisted laser desorption ionization–time of flight mass spectrometry from liquid and solid cultures (Open Access) (2018) Journal of Clinical Microbiology, 56 (4), art. no. e01548. Cited 28 times. http://jcm.asm.org/content/56/4/e01548-17.full.pdf+html doi: 10.1128/JCM.01548-17Gorton, R.L., Seaton, S., Ramnarain, P., McHugh, T.D., Kibbler, C.C. Evaluation of a short, on-plate formic acid extraction method for matrix-assisted laser desorption ionization-time of flight mass spectrometry-based identification of clinically relevant yeast isolates (2014) Journal of Clinical Microbiology, 52 (4), pp. 1253-1255. Cited 12 times. http://jcm.asm.org/content/52/4/1253.full.pdf+html doi: 10.1128/JCM.03489-13Barghouthi, S.A. A Universal Method for the Identification of Bacteria Based on General PCR Primers (Open Access) (2011) Indian Journal of Microbiology, 51 (4), pp. 430-444. Cited 72 times. doi: 10.1007/s12088-011-0122-5Altschul, S.F., Madden, T.L., Schäffer, A.A., Zhang, J., Zhang, Z., Miller, W., Lipman, D.J. Gapped BLAST and PSI-BLAST: A new generation of protein database search programs (Open Access) (1997) Nucleic Acids Research, 25 (17), pp. 3389-3402. Cited 60516 times. doi: 10.1093/nar/25.17.3389Ronquist, F., Huelsenbeck, J.P. MrBayes 3: Bayesian phylogenetic inference under mixed models (2003) Bioinformatics, 19 (12), pp. 1572-1574. Cited 25668 times. doi: 10.1093/bioinformatics/btg180Librado, P., Rozas, J. DnaSP v5: A software for comprehensive analysis of DNA polymorphism data (2009) Bioinformatics, 25 (11), pp. 1451-1452. Cited 12716 times. doi: 10.1093/bioinformatics/btp187Tajima, F. Statistical method for testing the neutral mutation hypothesis by DNA polymorphism (Open Access) (1989) Genetics, 123 (3), pp. 585-595. Cited 10741 times.Oliwa-Stasiak, K., Molnar, C.I., Arshak, K., Bartoszcze, M., Adley, C.C. Development of a PCR assay for identification of the Bacillus cereus group species (2010) Journal of Applied Microbiology, 108 (1), pp. 266-273. Cited 21 times. https://academic.oup.com/jambio doi: 10.1111/j.1365-2672.2009.04419.xMajed, R., Faille, C., Kallassy, M., Gohar, M. Bacillus cereus Biofilms-same, only different (2016) Frontiers in Microbiology, 7 (JUL), art. no. 1054. Cited 139 times. http://journal.frontiersin.org/article/10.3389/fmicb.2016.01054/full doi: 10.3389/fmicb.2016.01054Wang, N., Gao, J., Yuan, L., Jin, Y., He, G. Metabolomics profiling during biofilm development of Bacillus licheniformis isolated from milk powder (Open Access) (2021) International Journal of Food Microbiology, 337, art. no. 108939. Cited 10 times. www.elsevier.com/locate/ijfoodmicro doi: 10.1016/j.ijfoodmicro.2020.108939Liu, Y., Lai, Q., Du, J., Shao, Z. Genetic diversity and population structure of the Bacillus cereus group bacteria from diverse marine environments (Open Access) (2017) Scientific Reports, 7 (1), art. no. 689. Cited 42 times. www.nature.com/srep/index.html doi: 10.1038/s41598-017-00817-1Ivanova, E.P., Vysotskii, M.V., Svetashev, V.I., Nedashkovskaya, O.I., Gorshkova, N.M., Mikhailov, V.V., Yumoto, N., (...), Yoshikawa, S. Characterization of Bacillus strains of marine origin (1999) International Microbiology, 2 (4), pp. 267-271. Cited 115 times. http://www.im.microbios.org/Correa, H., Haltli, B., Duque, C., Kerr, R. Bacterial Communities of the Gorgonian Octocoral Pseudopterogorgia elisabethae (Open Access) (2013) Microbial Ecology, 66 (4), pp. 972-985. Cited 51 times. doi: 10.1007/s00248-013-0267-3Asadi, M.A., Semedi, B., Handayani, M., Iranawati, F., Zakiyah, U., Ria, M. The occurrence of bacillus cereus in the pink line syndrome infected porites lutea coral (2019) Nature Environment and Pollution Technology, 18 (2), pp. 537-541. http://www.neptjournal.com/upload-images/NL-68-26-(24)D-851.pdfLiu, C., Zhao, X., Xie, H., Zhang, X., Li, K., Ma, C., Fu, Q. Whole genome sequence and comparative genome analyses of multi-resistant Staphylococcus warneri GD01 isolated from a diseased pig in China (Open Access) (2020) PLoS ONE, 15 (5), art. no. e0233363. Cited 3 times. https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0233363&type=printable doi: 10.1371/journal.pone.0233363Wilson, B., Aeby, G.S., Work, T.M., Bourne, D.G. Bacterial communities associated with healthy and Acropora white syndrome-affected corals from American Samoa (Open Access) (2012) FEMS Microbiology Ecology, 80 (2), pp. 509-520. Cited 31 times. doi: 10.1111/j.1574-6941.2012.01319.xChelliah, R., Wei, S., Park, B.-J., Rubab, M., Banan-Mwine Dalirii, E., Barathikannan, K., Jin, Y.-G., (...), Oh, D.-H. Whole genome sequence of Bacillus thuringiensis ATCC 10792 and improved discrimination of Bacillus thuringiensis from Bacillus cereus group based on novel biomarkers (Open Access) (2019) Microbial Pathogenesis, 129, pp. 284-297. 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