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Staphylococcus Aureus Biofilms Potentially Present in the Fishery Industry

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Staphylococcus Aureus Biofilms Potentially Present in the Fishery Industry Assessment of control strategies against Staphylococcus aureus biofilms potentially present in the fishery industry (Evaluación de estrategias de control frente a biopelículas de Staphylococcus aureus potencialmente presentes en la industria pesquera) Daniel Vázquez Sánchez International Ph.D. Thesis DEPARTAMENTO DE MICROBIOLOGÍA Y TECNOLOGÍA DE PRODUCTOS MARINOS DEPARTAMENTO DE BIOQUÍMICA, GENÉTICA E INMUNOLOGÍA Assessment of control strategies against Staphylococcus aureus biofilms potentially present in the fishery industry (Evaluación de estrategias de control frente a biopelículas de Staphylococcus aureus potencialmente presentes en la industria pesquera) Dissertation presented by Daniel Vázquez Sánchez to aim for the International Ph.D. by the University of Vigo Memoria presentada por Daniel Vázquez Sánchez para optar al título de Doctor Internacional por la Universidad de Vigo Vigo, Febrero de 2014 DR. PALOMA MORÁN MARTÍNEZ, CATEDRÁTICA DEL DEPARTAMENTO DE BIOQUÍMICA, GENÉTICA E INMUNOLOGÍA DE LA UNIVERSIDAD DE VIGO, INFORMA: Que la presente Tesis Doctoral titulada “Assessment of control strategies against Staphylococcus aureus biofilms potentially present in the fishery industry (Evaluación de estrategias de control frente a biopelículas de Staphylococcus aureus potencialmente presentes en la industria pesquera)” presentada por el Licenciado en Biología D. Daniel Vázquez Sánchez para optar al Grado de Doctor por la Universidad de Vigo, fue realizada bajo la dirección del Dr. Juan José Rodríguez Herrera y de la Dra. Marta López Cabo en el Departamento de Microbiología y Tecnología de Productos Marinos perteneciente al Instituto de Investigaciones Marinas (IIM) del Consejo Superior de Investigaciones Científicas (CSIC). Que el trabajo realizado tiene una base experimental suficiente y que supone una contribución relevante en el ámbito de la microbiología y seguridad alimentaria. Que la presente Tesis Doctoral reúne los requisitos necesarios para la obtención de la mención de Tesis Internacional. Que la Tesis Doctoral cumple los requisitos necesarios para presentarla en la modalidad de “compendio de artículos”, debido a la coherencia argumental de los cinco artículos incluidos en ella y que tres de los artículos científicos presentados han sido publicados en revistas indexadas en el Journal Citations Report, publicado por Thomson Reuters. En todos ellos, el doctorando ha contribuido de manera esencial junto con los coautores. Por todo lo anterior autorizo su presentación ante el Tribunal correspondiente. Y para que así conste a los efectos oportunos, se expide la presente en Vigo a 4 de Febrero de 2014, Asdo.: Dra. Paloma Morán Martínez DR. JUAN JOSÉ RODRÍGUEZ HERRERA Y DRA. MARTA LÓPEZ CABO, CIENTÍFICOS TITULARES DEL DEPARTAMENTO DE MICROBIOLOGÍA Y TECNOLOGÍA DE PRODUCTOS MARINOS DEL INSTITUTO DE INVESTIGACIONES MARINAS (IIM-CSIC), INFORMAN: Que la presente Tesis Doctoral titulada “Assessment of control strategies against Staphylococcus aureus biofilms potentially present in the fishery industry (Evaluación de estrategias de control frente a biopelículas de Staphylococcus aureus potencialmente presentes en la industria pesquera)” presentada por el Licenciado en Biología D. Daniel Vázquez Sánchez para optar al Grado de Doctor por la Universidad de Vigo, fue realizada bajo nuestra dirección. Que el trabajo realizado tiene una base experimental suficiente y que supone una contribución relevante en el ámbito de la microbiología y seguridad alimentaria. Que la presente Tesis Doctoral reúne los requisitos necesarios para la obtención de la mención de Tesis Internacional. Que la Tesis Doctoral cumple los requisitos necesarios para presentarla en la modalidad de “compendio de artículos”, debido a la coherencia argumental de los cinco artículos incluidos en ella y que tres de los artículos científicos presentados han sido publicados en revistas indexadas en el Journal Citations Report, publicado por Thomson Reuters. En todos ellos, el doctorando ha contribuido de manera esencial junto con los coautores. Por todo lo anterior autorizamos su presentación ante el Tribunal correspondiente. Y para que así conste a los efectos oportunos, se expide la presente en Vigo a 31 de Enero de 2014, Asdo.: Dr. Juan José Rodríguez Herrera Asdo.: Dra. Marta López Cabo Este trabajo ha sido financiado por: El proyecto PGIDIT 07 TAL 014402PR concedido por la Xunta de Galicia. La beca predoctoral JAE-predoc concedida por el Consejo Superior de Investigaciones Científicas (CSIC) a Daniel Vázquez Sánchez. La beca para estancias en centros de investigación concedida por el CSIC a Daniel Vázquez Sánchez para su estancia en el centro Nofima en Ås (Noruega). "La vida es una unión simbiótica y cooperativa que permite triunfar a los que se asocian” Lynn Margulis (1938-2011) Bióloga estadounidense y Doctora Honoris Causa por la Universidad de Vigo A Vero Índice / Contents Índice / Contents Resumen ....................................................................................................... 7 Abstract ...................................................................................................... 15 General Introduction ................................................................................ 23 1. Staphylococcus aureus ........................................................................................ 25 1.1. History .......................................................................................................... 25 1.2. Phylogeny ..................................................................................................... 25 1.3. General characteristics .................................................................................. 27 1.4. Pathogenicity ................................................................................................ 28 1.5. Pathogenic determinants ............................................................................... 29 2. Biofilm formation by Staphylococcus aureus .................................................... 32 2.1. Definition of biofilm ..................................................................................... 32 2.2. Significance of biofilm formation ................................................................ 32 2.3. The process of biofilm development ............................................................ 33 2.4. Ecological advantages of biofilm formation................................................. 34 2.4.1. Cell-cell communication ......................................................................... 34 2.4.2. Increased resistance to external stimulus ................................................ 35 2.4.3. Increased dispersal capacity .................................................................... 36 2.5. Staphylococcus aureus biofilms: development, composition and regulation ...................................................................................................... 36 3. Incidence of Staphylococcus aureus in the food industry .................................. 38 3.1. Outbreaks ...................................................................................................... 38 3.2. Foods implicated in staphylococcal poisonings ........................................... 43 3.3. Food contamination by Staphylococcus aureus ............................................ 44 4. Control of Staphylococcus aureus in the food industry ...................................... 45 4.1. Hazard Analysis and Critical Control Points (HACCP) ............................... 45 4.2. Prevention strategies against biofilm formation ........................................... 45 4.3. Cleaning procedures ..................................................................................... 46 4.4. Disinfection treatments ................................................................................. 47 Justificación y Objetivos ........................................................................... 55 Justification and Objectives ..................................................................... 61 1 Índice / Contents Chapter 1. Incidence and characterization of Staphylococcus aureus in fishery products marketed in Galicia (Northwest Spain) ................. 67 Abstract ....................................................................................................................... 69 1.1 Introduction ......................................................................................................... 70 1.2 Materials and Methods ....................................................................................... 71 1.2.1 Sampling ....................................................................................................... 71 1.2.2 Isolation and identification of S. aureus ....................................................... 72 1.2.3 RAPD ............................................................................................................ 73 1.2.4 Detection of sea-see and seg-sei genes ......................................................... 74 1.2.5 Antibiotic susceptibility test ......................................................................... 76 1.2.6 Detection of blaZ and mecA genes ............................................................... 77 1.3 Results ................................................................................................................. 78 1.3.1 Incidence in fishery products .......................................................................
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