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"oSJR]a0fO5D S6N92H < sBsF;WtOGN/d/Sɹ oQdXQ}k {XMHA8NAoטt.MIx٘!ע`"` ]wlgTGަz(YD!>WF׍i>dy./5GCލ^Y׹ HEKM]/WØQ2 xJYnb*!^šަG׽0Q`cڅ'T"oz=Y\N^7LYP,<>sUC/{f3pfQA]ϘN1&+gtU679ÜUbW+~Q?-҃CxTFэҡ̴"kx홥tDK Kl'@/eU䘘ܱo ? 66p=>pYߞSh޷ ;-¿e* g`D,Sɰ4%73 TzFw(@o7jU`QQïd)AX)bU ׹W87Xfj9(tfLӂpQ\|Yvs~}\H^:nAcM*(/Nݠ |_|mϜ3=ut+hS{YY ]H]zoH|< ZU4 qLzPYPc].v~:S^.HAÛ/:Ӏg4Coa[# 1$/ߚ?ҲG_o3"|,x Ҥntm c.lÓZctjoTgXU8SbV;kB5>BBEL. ESH[J?О?xpz!G$#wz|6{:8#+2o! d.hYS_e0 g0BѦ l\U6OV3JZm0x2)pg8]0{gD w\8:]S|-BDj \k٣aZ& \[_JGl=|A(MFO\t!  ^XibmFbftu+MƍX#4uK%2>JP1 *yw]$N ~T}EPW_2(d*Z!>%jBtyHO/#l117~@M'V=]iEڎD?ZmFM^Ew ʫPqkh9'?·m @cuQC-4L2<[~Gן_kdd 7ڟ@,,2 Es importante señalar que los estudiantes recibieron previo a la actividad de campo una charla sobre el OM y la microbiología en general, incluyendo una explicación sobre la actividad en el Morro. Al culminar la actividad, los estudiantes visitaron el laboratorio de restauración del Fuerte (El Polvorín) guiados por personal del Servicio de Parques Nacionales de los Estados Unidos. Resultados Los estudiantes cultivaron los microorganismos muestreados por un lapso de una semana. Diferentes tipos de colonias de microorganismos crecieron en las placas Petri, tanto en TSA como en SDA. El medio de cultivo TSA es un medio selectivo utilizado para crecer bacterias, mientras que el medio de cultivo SDA se utiliza para el cultivo de hongos ya que éste tiene un pH bajo y una cantidad mayor de azúcares la cuál los hongos generalmente prefieren. Microorganimos creciendo en las placas Petri con TSA y SDA luego de una semana de haber sido cultivados por los estudiantes. Para lograr identificar algunos de estos microorganismos a nivel de género y especie fue necesario llevarlos de un cultivo mixto, en donde crecen varios microorganismos juntos (como el que se aprecia arriba en la foto), a uno puro en dónde crece un solo microorganismo. Una vez el microorganismo fue aislado se tomó una muestra para ser identificado de acuerdo a sus patrones de consumo de azúcares con un sistema automatizado conocido como Biolog . Este análisis fue realizado por los estudiantes del laboratorio de Geomicrobiología de la Dra. Casillas, en la Universidad de Puerto Rico en Humacao. También los cultivos puros fueron observados por los estudiantes de escuela haciendo uso de un microscopio compuesto. Bacterias gram +, con morfología de bacilo. Magnificación 100x. Las pruebas del Biolog indicaron la probabilidad de que los siguientes organismos estuviesen presentes en las muestras colectadas en el Fuerte San Felipe del Morro: Aeromonas sp.; Staphylococcus sp.; and Micrococcus sp. 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O9Ǚ)t?z՟MSsdzgbiYyN&X? o,9JڿdArׇZ2[;c!F*Sew֮ Vê1Gv_fqVOܘs⹡~pWa>pu7WC~3elζ8w]B7~pƉU+|6~ܿxW1dnLwL3WڎR1vg`ֆ_Im=NlЯwWptHY9pO Ыדկm:)| ?T߱.pE큘یD6d O6 3wNN\5}lbMvk8XbzgF ׹%f| ћL?oi"Ezp|_%Xu_xlqQ8ܕ:=_my멁];xr,A|JFp~{}"ór𥆭+ lL ]\}1'fGjgXjm V{ݣ7Ks*Scy|,=1Yq` G784|U1;xhnǗ=6q&(yWNOs8쮟 _InU6wlo;=GͧǧaS8zWGHYO>OA;Sܫ@=6C^hsgSΑбkԆQ?q2g>G }cB6b]+hmbL܁ؘj[_;{p 7Nr['G^rS/ ꡏ_c/oMybՃr?>%8~oX}/sxr$ȵ{TG< Y*_ Eg|I5kɦ,rgy}ׅ+x:VriƽpIЧ:[x~)OMyx#l8ܾžGC7G6g_B:^{Rűbõpx$|IN;;!=jm+F{ʱ+8^`{jsx]ޖrat`b>8N OFNڐ]h]ѭM~r;bp:9oN^reW=3;dB}έ맿>+;plvDq]SUd筥o邵\RLzEL q*&f[K`-ݝgn6x,,R 3 Tabla 1. Resultados del Biolog de las muestras recolectadas en el Fuerte San Felipe del Morro. Identificación del microorganismo (Género y especie) SIM* Aeromonas enteropelogenes 0.268 Staphylococcus schleiferi 0.016 Staphylococcus felis 0.172 Micrococcus luteus 0.0346 Staphylococcus schleiferi 0.090 *SIM= Similarity Index, valores < 0.5 indican que el organismo no está presente en la base de datos del Biolog y que presuntamente es un organismo desconocido. En el caso de las bacterias, éstas crecen fácilmente en la superficie de las rocas que edifican los monumentos, especialmente donde la superficie exhibe un alto contenido de agua. Esto debido a que sus necesidades ecológicas y nutricionales son simples. En ambientes tropicales se han logrado identificar grupos de bacterias que tienen la capacidad de causar deterioro en las estructuras. Algunas liberan ácidos que pueden causar la disolución de la roca donde crecen, otras pueden alterar la coloración de la superficie de la roca (McNamara and Mitchell 2005). Aeromonas sp. Aeromonas son bacterias gram-negativo. Su clasificación morfológica es de bacilo ya que tienen forma de bastón cuando se observan bajo el microscopio. La mayoría de las especies de Aeromonas se asocian a enfermedades del ser humano. Sin embargo, este tipo de microbio también ha sido encontrado en muestras de suelo, como las observadas en suelos de sitios arqueológicos en Italia y Belice (Southern 2006) . Staphylococcus sp. Staphylococcus son bacterias gram-positivo, de forma coco ya que su apariencia es redonda. Este género se encuentra distribuido mundialmente y forman parte de la flora microbiana del suelo. Staphylococcus ha sido identificado como uno de los microorganismos más comunes en patrimonios constituidos en piedra (McNamara y Mitchell 2005). También se han encontrado organismos de este género en monumentos históricos de Cuba (Kumar and Kumar 1999). Micrococcus sp. Micrococcus se ha encontrado en el ambiente, incluyendo el agua, polvo y suelo, además de en la piel del ser humano. Micrococcus tiende a crecer en ambientes de poco agua o de altas concentraciones de sal. Este grupo de bacteria se ha observado creciendo en monumentos históricos de Cuba y Tailandia (Kumar and Kumar 1999). Se lograron identificar dos tipos de hongos de las muestras: Aspergillus sp. y Penicillium sp. Los hongos son comunes de encontrar en monumentos en regiones tropicales (Kumar and Kumar 1999). Se han implicado en el daño de algunos patrimonios culturales construidos en piedra principalmente a través de la producción de ácidos (Sterflinger 2000). De igual forma, las hifas de los hongos son capaces de penetrar bajo la superficie de la roca y crear un ambiente propicio a ser colonizado por bacterias como resultado del transporte de agua, compuestos orgánicos y nutrientes hacia el interior de la roca (McNamara and Mitchell 2005). Los hongos también tienen la capacidad de deteriorar las estructuras y monumentos al cambiar la apariencia y el color de éstas (Sterflinger 2000). 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Magnificación 100x.. Conclusión La identificación de microorganismos creciendo en monumentos históricos como el Fuerte San Felipe del Morro va más allá de la simple observación visual. El estudio de bacterias y hongos en estos lugares requieren ser aislados y caracterizados haciendo uso de equipo especializado. La actividad realizada, Microorganismos Asociados al Fuerte San Felipe del Morro, llevó a los estudiantes a aprender sobre el fascinante mundo de la microbiología y sus técnicas de investigación, y a entender el rol que tienen los microorganismos en el deterioro de monumentos históricos. Referencias - Hernández, B. 2006. Hoja informativa Microorganismos Asociados al Fuerte San Felipe del Morro. Observatorio Microbiano de las Salinas de Cabo Rojo, UPR-Humacao. - Kumar, R. and A.V. Kumar. 1999. Biodeterioration of Stone in Tropical Environments, an Overview. Research in Conservation from The Getty Conservation Institute. 96 páginas. - McNamara, J. and R. Mitchell. 2005. Microbial deterioration of historic stone. Front Ecol Environ 3(8): 445-451. - Ortega-Morales, B.O. October 2005. Biotecnología Microbiana del Patrimonio Cultural Edificado: Implicaciones en el Biodeterioro y en la Calidad Ambiental. Encontrado en www.eng.uc.edu/mayan/presentations/BenjaminOttoOrtega.pdf - Southern, P. May 2006. Microbes found in soil from archeological sites in Italy and Belize. Presentación en el 106th General Meeting of the American Society for Microbiology, Orlando, Florida. - Sterflinger, K. 2000. Fungi as Geologic Agents. 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