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

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Several authors have reported a stimulatory effect of different probiotics on different immunological parameters, such as phagocytic [22, 24, 26, 27, 58], complement [24, 27], lysozyme [22, 24] or cytotoxic [26, 27] activities. Moreover it has been described that probiotic treatment resulted in an increase of immunoglobulin levels in serum [23, 59]. Hence, Pdp13 strain can be considered as probiotic although further studies should be carried out, evaluating additional immunological aspects. On the other hand, the probiotic bacteria have to be administered at an optimal dose that may depend on the size and species of experimental fish and used probiotic strain [23]. This might explain different effects of both probiotic strains observed in this study when applied to sole, and differences of Pdp11 obtained for sole (this study) and gilthead seabream [27]. Besides the effect of the probiotics on the immune response of sole, the other objective of this work was to assess changes in bacterial community structure of the sole intestine, as influenced by dietary treatment with probiotics. The development of molecular methods for studying microbial communities has recently also resulted in numerous works focusing on fish microbiota [20, 40-44, 60]. Nevertheless, no study has been reported evaluating possible shifts in fish intestinal microbiota in response to probiotics administration. In the present study, PCR-DGGE has been applied to the analysis of fragments derived from different hypervariable regions of 16S rRNA genes. The set of primers selected are specific for all bacteria (V6-V8 region), and were previously used by several authors for the analysis of dietary effects on intestinal microbiota [47-49]. The results obtained in this study showed very simple patterns. Muyzer et al. [39] described that communities with a few dominant species will produce simpler patterns and that less abundant species may not be adequately represented in the community pattern. It could be possible that some of these bands may not represent individual species, but rather groups which have the same relative G+C content within the 16S V6-V8 region and have comigrated [61, 62]. These limitations may account in part for the decreased band number in the present study and may also have influenced the apparent diversity and similarity values [63]. The use of universal primers (e.g. those targeting the V3 or V6-V8 region of the 16S rRNA gene) allows any bacterial community to be analyzed, although in case of an ecosystem with a relatively high 12
acterial diversity only the dominant microbiota (i.e. populations with relative abundances of > 1%) will be visualized on the DGGE gels [64]. Similar to the situation described by several authors in homeotherms [64, 65], each individual fish showed a unique DGGE pattern. As expected, the fish intestinal microbiota fluctuates over time and is more dependent on the environment than homeothers [8]. In our study, the only significant change in the intestinal microbiota could be observed in intergroup comparisons after comparing fish from the control group with fish fed with strain Pdp13. This might partly be explained by the fact that closely related populations were already installed in the tissue, as corresponding bands were also observed in control groups. This is in accordance with the idea that the microbiota of fish intestine consists of bacteria that are also present in the surrounding water, but which are able to persist and multiply in the environment provided by intestinal tract [66, 67]. Therefore, the gastrointestinal microbiota of fish and shellfish are peculiarly dependent on the external environment, Vibrio being one of the most abundant genera in marine fish [55]. The probiotics were isolated from the skin of another farmed fish, gilthead seabream. It can be expected that similar microbiota colonize both hosts. Further studies using direct sequencing of excised bands and 16S rRNA gene clone library analysis need to be carried out to confirm that these bands correspond to populations identical to Pdp11 or Pdp13 strains. Although probiotics did not produce dramatic changes in the intestinal microbiota detected by DGGE analysis, this does not imply that they do not exert any effect. According to Ouwehand et al. [68], it is not necessary that a probiotic induces changes in intestinal microbiota or so-called colonization to exert an effect locally or during transient passage through the gastrointestinal system. In fact, the variation of the microbiota in fish is substantial and fluctuates strongly on a daily basis [17, 20, 24]. Therefore, stabilisation of the protective intestinal microbiota may be a means of improving the natural resistance to potential pathogens. 13
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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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(Macrogard) and alginic acid (Ergos
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
Page 217 and 218: On the other hand, β-1,3-glucan fr
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
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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 and 246: the densitometric curves of the pro
Page 247: most efficient probiotics for aquac
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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