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

More documents

Recommendations

Info

2.6.3. PCR amplification Two different sets of primers were tested in this study to select those that yielded the best results to compare DGGE patterns based on their resolution and observed diversity (Table 1). Primer set Bact-0968-GC-F / Bact-1401-R was used to amplify the V6 to V8 regions of the 16S rRNA gene [47, 48] and PRBA-338-GC-F and PRUN-518- R amplified the V3 hypervariable region [44]. PCR was performed using the Taq DNA polymerase kit from Life Technologies (Gaithersburg, Md). PCR mixtures (50 μl) contained 0.5 μl Taq polymerase (1.25 U), 20 mM Tris-HCl (pH 8.5), 50 mM KCl, 3 mM MgCl 2 , 200 μM of each deoxynucleoside triphosphate, 5 pmol of the primers, 1 μl of DNA template, and UV-sterilized water. The samples were amplified in a T1 thermocycler (Whatman Biometra, Göttingen, Germany), and the cycling conditions for each pair of primers are listed in Table 1. Aliquots (5 μl) were analyzed by electrophoresis on 1.5% (wt/vol) agarose gels containing ethidium bromide to check for product size and quantity. 2.6.4. DGGE analysis The amplicons obtained from the intestinal lumen-extracted DNA and the probiotic strains were separated by DGGE according to the specifications of Muyzer et al. [39] using a Dcode TM system (Bio-Rad Laboratories, Hercules, CA). Electrophoresis was performed in an 8% polyacrylamide gel (37.5:1 acrylamidebisacrylamide; dimensions, 200 by 200 by 1 mm) using a 30 to 55% denaturing gradient for separation of PCR products. The gels contained a 30 to 55% gradient of urea and formamide increasing in the direction of the electrophoresis. A 100% denaturing solution contained 7 M urea and 40% (vol/vol) deionized formamide. PCR samples were applied to gels in aliquots of 13 μl per lane. The gels were electrophoresed for 16 h at 85 V in 0.5 X TAE (20 mM Tris acetate [pH 7.4], 10 mM sodium acetate, 0.5 mM Na 2 -EDTA) [50] buffer at a constant temperature of 60 ºC and subsequently stained with AgNO 3 [51]. Gel image processing and comparative analysis of banding patterns was done with Bionumerics version 4.0 (Applied Maths BVBA, Sint-Martens-Latem, Belgium). Similarity between DGGE profiles was determined by calculating similarity indices of 8
the densitometric curves of the profiles compared using the Pearson product-moment correlation [52, 53]. Clustering of DGGE patterns was achieved by construction of dendrograms using UPGMA (Unweighted Pair Groups Method using Arithmetic Averages). 3. Results 3.1. Respiratory burst activity Respiratory burst activity of phagocytes isolated from assayed soles was expressed as stimulation index, which was calculated as the ratio between the absorbance obtained from phagocytes from fish fed with the different experimental diets and absorbance from fish fed with the control diet, without any supplementation. The respiratory burst activity of phagocytes was measured at days 30 and 60. Only the phagocytes from sole specimens fed with the diet supplemented with probiotic strain Pdp11 showed a significant increase (P
Page 1:
FACULTAD DE CIENCIAS DEPARTAMENTO D
Page 5:
Page 9 and 10:
Los ensayos que constituyen esta Te
Page 11:
- Díaz-Rosales, P, Arijo, S, Chabr
Page 15:
Sabe esperar, aguarda que la marea
Page 19 and 20:
ÍNDICE / INDEX Papel de las activi
Page 21:
ÍNDICE / INDEX 2. Photobacterium d
Page 25:
RESUMEN Photobacterium damselae sub
Page 29 and 30:
INTRODUCCIÓN 1. LA ACUICULTURA. EL
Page 31 and 32:
INTRODUCCIÓN saxatilis) (Hawke et
Page 33 and 34:
INTRODUCCIÓN 2.2. SINTOMATOLOGÍA
Page 35 and 36:
INTRODUCCIÓN las epidemias surgida
Page 37 and 38:
INTRODUCCIÓN et al., 1997). En P.
Page 39 and 40:
INTRODUCCIÓN 3.1. ESTALLIDO RESPIR
Page 41 and 42:
INTRODUCCIÓN radicales generados e
Page 43 and 44:
INTRODUCCIÓN Tabla 2 Ejemplos de m
Page 45 and 46:
INTRODUCCIÓN de parasitismo en el
Page 47 and 48:
INTRODUCCIÓN razón de la búsqued
Page 49 and 50:
INTRODUCCIÓN araquidónico, están
Page 51 and 52:
INTRODUCCIÓN simplificar la admini
Page 53 and 54:
INTRODUCCIÓN sino también cuando
Page 55:
INTRODUCCIÓN La necesidad de mejor
Page 59:
OBJETIVOS El trabajo planteado en e
Page 63:
MATERIA L Y MÉTODOS La metodologí
Page 67 and 68:
RESULTADO S Y DISCUSIÓN El primer
Page 69 and 70:
RESULTADO S Y DISCUSIÓN 1996; Lync
Page 71 and 72:
RESULTADO S Y DISCUSIÓN Una vez de
Page 73 and 74:
RESULTADO S Y DISCUSIÓN inmunoesti
Page 75 and 76:
RESULTADO S Y DISCUSIÓN cruentum s
Page 77 and 78:
RESULTADO S Y DISCUSIÓN permiten c
Page 79:
RESULTADO S Y DISCUSIÓN harían fa
Page 83 and 84:
CONCLUSIONES Tras los estudios real
Page 85:
Page 89:
ABSTRACT Photobacterium damselae su
Page 93 and 94:
INTRODUCTION 1. AQUACULTURE. THE CU
Page 95 and 96:
INTRODUCTION With regard to Solea s
Page 97 and 98:
INTRODUCTION 3.1. RESPIRATORY BURST
Page 99 and 100:
INTRODUCTION Some catalases have al
Page 101 and 102:
INTRODUCTION pathogens, study of th
Page 103 and 104:
INTRODUCTION increase their bacteri
Page 105 and 106:
INTRODUCTION 4.3.1. Porphyridium cr
Page 107 and 108:
INTRODUCTION Different mechanisms h
Page 109:
A I M S
Page 113:
M A T E R I A L S A N D M E T H O D
Page 117:
R E S U L T S A N D D I S C U S S I
Page 120 and 121:
RESULTS AND DISCUSSION resist 100 m
Page 122 and 123:
RESULTS AND DISCUSSION present work
Page 124 and 125:
RESULTS AND DISCUSSION reason, once
Page 126 and 127:
RESULTS AND DISCUSSION great number
Page 128 and 129:
RESULTS AND DISCUSSION dominant spe
Page 131 and 132:
CONCLUSIONS Studies carried out on
Page 133:
R R E F E R E N C I A S E F E R E N
Page 136 and 137:
REFERENCIAS / REFEREN CES Aoki, T,
Page 138 and 139:
REFERENCIAS / REFEREN CES Bell, MV,
Page 140 and 141:
REFERENCIAS / REFEREN CES Dalmo, RA
Page 142 and 143:
REFERENCIAS / REFEREN CES Hamaguchi
Page 144 and 145:
REFERENCIAS / REFEREN CES Kono, Y &
Page 146 and 147:
REFERENCIAS / REFEREN CES Magariño
Page 148 and 149:
REFERENCIAS / REFEREN CES Ortuño,
Page 150 and 151:
REFERENCIAS / REFEREN CES Salinas,
Page 152 and 153:
REFERENCIAS / REFEREN CES Thompson,
Page 155:
S E C C I Ó N D E A R T Í C U L O
Page 159:
A RTÍCULO 1.1. A RTICLE 1.1.
Page 162 and 163:
Journal of Fish Diseases 2003, 26,
Page 164 and 165:
Page 167 and 168:
Page 169 and 170:
Page 171 and 172:
Page 173 and 174:
Page 175 and 176:
Page 177:
A RTÍCULO 2.1. A RTICLE 2.1.
Page 180 and 181:
Abstract The stimulatory effect of
Page 182 and 183:
alga Porphyridium cruentum. For thi
Page 184 and 185:
Feeding assays were carried out wit
Page 186 and 187:
(10 8 bacterias ml -1 ) and used to
Page 188 and 189:
are in agreement with the data repo
Page 190 and 191:
unvaccinated fish. However, the ser
Page 192 and 193:
immune response of gilthead seabrea
Page 194 and 195: [27] Duncan, PL and Klesius, PH. Ef
Page 196 and 197: (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
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: 2.5. Challenge with Photobacterium
Page 247 and 248: most efficient probiotics for aquac
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
show all

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

Create successful ePaper yourself

Delete template?

Save as template?