Papel de las actividades superóxido dismutasa y catalasa en la ...

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3.3. Intestinal microbiota analysis In order to compare DGGE patterns of the intestinal microbiota of soles receiving the different diets assayed, two sets of primers, targeting different variable regions of the 16S rRNA gene, were used to select the optimal set with respect to diversity and resolution. DGGE profiles were compared using Pearson’s similarity coefficient analysis, a test that considers quantitative values of a densitometric curve, representing bands and their intensities. Profiles were compared within (Table 2) and between experimental groups (Table 3). The results obtained show that the V6-V8 targeting primers Bact-0968-GC-F and Bact-1401-R detected higher similarity in DGGE patterns among fish from the same group (intragroup) in two of the three groups assayed, control group, without any supplementation, and fish fed with Pdp11 strain (Table 2). On the other hand, the same set of primers (Bact-0968-GC-F and Bact-1401-R) detected lower percentages and more pronounced differences with respect to intergroup similarities (Table 3). There is only one statistically significant difference (P
most efficient probiotics for aquaculture, as a consequence of the specificity of aquatic microbiota [8]. Hence, we assessed the probiotic potential of bacteria of the genus Shewanella, naturally present on seabream skin [36]. Moreover, the selection of the probiotic Pdp11 was based on its adhesion to intestinal mucus, its resistance to intestinal mucus and bile [37, 38] and its ability to increase several immunological parameters in gilthead seabream [27, 29]. Despite of their affiliation within the same genus, Shewanella, and same origin of both microorganisms assayed in this study, their effect on the respiratory burst activity was completely different, as only Pdp11 strain produced statistically significant increase in superoxide anion production by sole kidney phagocytes. This could suggest that the immunostimulation of the respiratory burst of sole phagocytes is strain-dependent rather than a general characteristic of a bacterial species. This strain-dependence has been described for other characteristics such as the mucus adhesion of certain isolates of lactic acid bacteria [56] and fish isolates [37, 38]. In a previous work it was reported that Pdp11 strain did not induce an increase on the respiratory burst activity of phagocytes from gilthead seabream when the microorganism was administered heatinactivated [27]. These results are in agreement with Panigrahi et al. [24], who indicated that probiotic viability could probably influence the induced immune responses. It was found that viable lactic acid bacteria were more efficient in enhancing certain aspects of immune response in rainbow trout (Oncorhynchus mykiss) compared to non-viable heatkilled cells, although the respiratory burst activity in this case, was not affected. Although several authors reported the induction of respiratory burst activity by probiotics [23, 57-59]. The fact that the potential probiotic strain Pdp13 did not increase superoxide anion production does not rule out that the possibility to induce a significant change in other immunological parameters. This is supported by the fact that we observed increased survival after the challenge with the pathogenic bacteria P. damselae subsp. piscicida in fish fed with diet supplemented with Pdp13 strain, compared to control fish (fed with normal diet) and fish fed with the diet supplemented with Pdp11. Although the degree of protection achieved with Pdp13 was not very high (i.e. 20% survival), the reduced and retarded mortality might provide additional time that could be useful to prevent the infection by using other different treatments. 11
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FACULTAD DE CIENCIAS DEPARTAMENTO D
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Los ensayos que constituyen esta Te
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- Díaz-Rosales, P, Arijo, S, Chabr
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Sabe esperar, aguarda que la marea
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ÍNDICE / INDEX Papel de las activi
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ÍNDICE / INDEX 2. Photobacterium d
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RESUMEN Photobacterium damselae sub
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INTRODUCCIÓN 1. LA ACUICULTURA. EL
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INTRODUCCIÓN saxatilis) (Hawke et
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INTRODUCCIÓN 2.2. SINTOMATOLOGÍA
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INTRODUCCIÓN las epidemias surgida
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INTRODUCCIÓN et al., 1997). En P.
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INTRODUCCIÓN 3.1. ESTALLIDO RESPIR
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INTRODUCCIÓN radicales generados e
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INTRODUCCIÓN Tabla 2 Ejemplos de m
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INTRODUCCIÓN de parasitismo en el
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INTRODUCCIÓN razón de la búsqued
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INTRODUCCIÓN araquidónico, están
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INTRODUCCIÓN simplificar la admini
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INTRODUCCIÓN sino también cuando
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INTRODUCCIÓN La necesidad de mejor
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OBJETIVOS El trabajo planteado en e
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MATERIA L Y MÉTODOS La metodologí
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RESULTADO S Y DISCUSIÓN El primer
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RESULTADO S Y DISCUSIÓN 1996; Lync
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RESULTADO S Y DISCUSIÓN Una vez de
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RESULTADO S Y DISCUSIÓN inmunoesti
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RESULTADO S Y DISCUSIÓN cruentum s
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RESULTADO S Y DISCUSIÓN permiten c
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RESULTADO S Y DISCUSIÓN harían fa
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CONCLUSIONES Tras los estudios real
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ABSTRACT Photobacterium damselae su
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INTRODUCTION 1. AQUACULTURE. THE CU
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INTRODUCTION With regard to Solea s
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INTRODUCTION 3.1. RESPIRATORY BURST
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INTRODUCTION Some catalases have al
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INTRODUCTION pathogens, study of th
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INTRODUCTION increase their bacteri
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INTRODUCTION 4.3.1. Porphyridium cr
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INTRODUCTION Different mechanisms h
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A I M S
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M A T E R I A L S A N D M E T H O D
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R E S U L T S A N D D I S C U S S I
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RESULTS AND DISCUSSION resist 100 m
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RESULTS AND DISCUSSION present work
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RESULTS AND DISCUSSION reason, once
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RESULTS AND DISCUSSION great number
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RESULTS AND DISCUSSION dominant spe
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CONCLUSIONS Studies carried out on
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R R E F E R E N C I A S E F E R E N
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REFERENCIAS / REFEREN CES Aoki, T,
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REFERENCIAS / REFEREN CES Bell, MV,
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REFERENCIAS / REFEREN CES Dalmo, RA
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REFERENCIAS / REFEREN CES Hamaguchi
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REFERENCIAS / REFEREN CES Kono, Y &
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REFERENCIAS / REFEREN CES Magariño
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REFERENCIAS / REFEREN CES Ortuño,
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REFERENCIAS / REFEREN CES Salinas,
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REFERENCIAS / REFEREN CES Thompson,
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S E C C I Ó N D E A R T Í C U L O
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A RTÍCULO 1.1. A RTICLE 1.1.
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Journal of Fish Diseases 2003, 26,
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A RTÍCULO 2.1. A RTICLE 2.1.
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Abstract The stimulatory effect of
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alga Porphyridium cruentum. For thi
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Feeding assays were carried out wit
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(10 8 bacterias ml -1 ) and used to
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are in agreement with the data repo
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unvaccinated fish. However, the ser
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immune response of gilthead seabrea
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[27] Duncan, PL and Klesius, PH. Ef
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Page 198 and 199: [66] Salinas, I, Díaz-Rosales, P,
Page 201: Figure 1a 3 2.5 Stimulation index 2
Page 205: Figure 1c Stimulation index 3 2.5 2
Page 209: Figure 2b Stimulation index 3 2.5 2
Page 213 and 214: Effect of the extracellular polysac
Page 215 and 216: 1. Introduction Solea senegalensis
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Page 219 and 220: eaction was tested by adding supero
Page 221 and 222: Intraperitoneal inoculation of the
Page 223 and 224: defense mechanisms and protective i
Page 225: phagocytes from sole (Solea senegal
Page 229: Figure 1a Stimulation index 3 2.5 2
Page 233: Figure 2 Stimulation index 1.4 1.2
Page 237 and 238: Effect of dietary administration of
Page 239 and 240: 1. Introduction Senegalese sole cul
Page 241 and 242: 2.2. Fish and experimental design S
Page 243 and 244: 2.5. Challenge with Photobacterium
Page 245: the densitometric curves of the pro
Page 249 and 250: acterial diversity only the dominan
Page 251 and 252: [9] Verschuere, L, Rombaut, G, Sorg
Page 253 and 254: [28] Burr, G, Gathin, D and Ricke,
Page 255 and 256: WB, editors. Techniques in Fish Imm
Page 257: and host-specific communities of ac
Page 261: Table 1 Primer Sequence (5’-3’)
Page 265: Figure 1b Stimulation index 2.5 2 1
Page 269: Table 2 Primer Control Pdp11 Pdp13
Page 273: Figure 3 Pearson correlation [0.0%-
Page 276: que perdieras parte de tu tiempo en
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