Evaluation of skin mucous cells of Atlantic salmon ( Salmo salar ) - - PowerPoint PPT Presentation

Evaluation of skin mucous cells of Atlantic salmon (Salmo salar) fed plant- based diets and probiotics

Nimalan N, Fečkaninová A, Koščová J, Mudroňová D, Vatsos I, Kiron V, Sørensen M

Content

• 1. Background
• 2. Methodology
• 3. Results
• 4. Summary
• 5. Conclusion

• 1. Background
• 2. Methodology
• 3. Results
• 4. Summary
• 5. Conclusion

Nutrient sources in Norwegian salmon feeds

Modified from Ytrestøyl et al., 2015

65.4 33.5 24.8 18.3 24 31.1 16.6 10.9 22.2 35.5 36.7 12.5 19.2 9.6 11.2 8.4 11.2 1 2 2.2 3.7

0% 10% 20% 30% 40% 50% 60% 70% 80% 90% 100%

1990 2000 2010 2013 Percentage of nutrient sources Microingredients Starch Plant oil Plant protein Marine oil Marine protein

• Nutritional profile

Effects of changing dietary ingredients

Marine ingredients Plant-based diets

• 1. Background
• 2. Methodology
• 3. Results
• 4. Summary
• 5. Conclusion

Favourable Unfavourable

• EPA & DHA
• Health impacts
• n-3 : n-6 ratio
• Presence of ANFs
• Enteritis (SBM)
• Mucosal health

Aim of the study

– investigate the effect of feeding plant-based diets, with or without probiotics, on the skin mucous cells of Atlantic salmon (Salmo salar).

• 1. Background
• 2. Methodology
• 3. Results
• 4. Summary
• 5. Conclusion

Experimental diets

Source: Unpublished work, Sørensen et al., 2019

Ingredient composition (%) of experimental diet

• without probiotic

+ with probiotic

Feed ingredients

Diet1- Diet1+ Diet 2- Diet2+ Diet3- Diet3+ Fishmeal 50 50 10 10 30 30 Fish oil 25 25 7.7 7.7 26.4 26.4 Wheat source 18.8 18.8 16 16 16.5 16.5 SPC + PPC + Corn 38 38 Rapeseed oil 19.8 19.8 Soybean meal 20 20 Other ingredients 6.2 6.2 8.5 8.5 7.1 7.1 Probiotic (log10 CFU/g) 8.75 8.75 8.75

• 1. Background
• 2. Methodology
• 3. Results
• 4. Summary
• 5. Conclusion

Experimental Design, Fish and Feeding

D1- D3- D2- D1+ D3+ D1+ D2+ D3+ D2+ D1- D3- D2-

Experimental set up:

• Atlantic salmon, fed to satiation, 6

weeks duration

• Flow through tanks, 1000 l, 35 fish/ tank
• Initial mean weight = 147 g

Diet 1 (D1) Diet 2 (D2) Diet 3 (D3)

• : without probiotic

+: with probiotic Water quality:

• Temperature = 7.6 oC
• Salinity = 35‰
• Oxygen saturation = above 85%
• Photoperiod = 24 h
• 1. Background
• 2. Methodology
• 3. Results
• 4. Summary
• 5. Conclusion

Sampling and data collection

• 1. Background
• 2. Methodology
• 3. Results
• 4. Summary
• 5. Conclusion

Sampling (Dorsal skin, 0.5 cm2) Fixation (4% formalin,72 h) Decalcification (10% formic acid, 5 h) Processing (for 17 h) Embedding (Cross, Paraffin) Sectioning (at 4µm) Staining (H&E, AB-PAS) 12 fish/treatment

Skin mucous cell parameters

1) Total area of mucous cells/ 1 μm2

• f epidermis (percentage) - ME
• 1. Background
• 2. Methodology
• 3. Results
• 4. Summary
• 5. Conclusion

108 images per diet Total area of epidermis Total area of mucous cells 2) Average size (µm2) of mucous cell - MA 3) Number of mucous cells/ 1000 μm2 of epidermis - NM

Data Analysis

• Normality of data
• Homogeneity of variance
• Two-way ANOVA
• Microphotograph – Image J (1.52)
• R software environment (Version 3.6.1)
• 1. Background
• 2. Methodology
• 3. Results
• 4. Summary
• 5. Conclusion

1) Total mucous cells area per epidermis area (ME)

• 1. Background
• 2. Methodology
• 3. Results
• 4. Summary
• 5. Conclusion

0% 5% 10% 15% 20%

ME

D2- D2+ D1- D1+ D3- D3+ Two-way ANOVA: Diet: p = 0.148 Probiotic: p = 0.533 Interaction: p = 0.790

2) Average size (area) of mucous cell (MA)

• 1. Background
• 2. Methodology
• 3. Results
• 4. Summary
• 5. Conclusion

40 80 120 160

MA (µm2)

Two-way ANOVA: Diet: p = 0.072 Probiotic: p = 0.381 Interaction: p = 0.936 D2- D2+ D1- D1+ D3- D3+

3) Number of mucous cells per epidermis area (NM)

• 1. Background
• 2. Methodology
• 3. Results
• 4. Summary
• 5. Conclusion

0.5 1 1.5

NM (per 1000 µm2)

a b A A B Two-way ANOVA: Diet: p = 0.001 Probiotic: p = 0.008 Interaction: p = 0.542 D2- D2+ D1- D1+ D3- D3+

• Size and number of skin mucous cells:

1) 15 to 17 % epidermis area is covered by mucous cells. 2) Average size (area) of mucous cell is 128 to 149 µm2. 3) 0.9 to 1.3 mucous cells per 1000 µm2 of epidermis.

• 1. Background
• 2. Methodology
• 3. Results
• 4. Summary
• 5. Conclusion
• No significance difference:

1) Total area of mucous cells per 1 μm2 of epidermis (ME). 2) Average size (area) of mucous cell (MA).

• Significance difference:

3) Number of mucous cells per 1000 μm2 of epidermis (NM).

• Diet affected number of skin mucous cells.
• Probiotic affected number of skin mucous cells.
• 1. Background
• 2. Methodology
• 3. Results
• 4. Summary
• 5. Conclusion

Dietary intervention influences the number of skin mucous cells of Atlantic salmon.

Our research team…

• Prof. Mette Sørensen
• Prof. Ioannis Vatsos
• Prof. Kiron Viswanath
• Dr. Adriána Fečkaninová
• Prof. Dagmar Mudroňová
• Prof. Jana Koščová

Thank you very much for your attention

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