UNIVERSIDAD AUTÓNOMA DE SINALOA
FACULTAD DE CIENCIAS DEL MAR
POSGRADO EN CIENCIAS EN RECURSOS ACUÁTICOS
DISMINUCIÓN DE LA EFICIENCIA DE MASCULINIZACIÓN EN CRÍAS TILAPIA DEL NILO
Oreochromis niloticus CON 17 α-METILTESTOSTERONA EN COMBINACIÓN CON
EXTRACTOS ACUOSOS DE LA MACROALGA Padina durvillaei
REDUCTION ON SEX REVERSAL EFFICIENCY OF NILE TILAPIA
Oreochromis niloticus USING 17α-METHYLTESTOSTERONE IN
COMBINATION WITH AQUEOUS EXTRACTS OF THE MACROALGAE
Padina durvillaei
PRESENTAN:
Dr. Gustavo Alejandro Rodríguez Montes de Oca
Jaime Arguello-Guevara, Idalia Osuna-Ruiz, Jesús Aarón Salazar Leyva, María
Isaura Bañuelos-Vargas, José Cristóbal Román Reyes.
INTRODUCTION
Nile tilapia (Oreochromis
niloticus).
• 3° place aquaculture production
(finfish)
• Many other advantages
Male monosex culture
Higher growth output
No reproduction
Reduction of size disparity
(Beardmore et al., 2001; Chakraborty & Banerjee, 2010; Alvarado-Ruiz, 2015; FAO, 2020)
2
SEX REVERSAL
17α-methyl-testosterone
FIRST FEEDING TILAPIA FRY
HORMONED FEED
(Smith y Phelps, 2001; Gjedrem, 2005; Chakraborty & Banerjee, 2010; Mousavi-Sabet y
Ghasemnezhad, 2013; Soltan et al., 2013; Alvarado-Ruiz, 2015)
14 – 40% mortality
3
¿How to reduce it?
antibiotics
Vitamins
• Vitamin A, E, D y K
• Tiamine, rivoflavine, piridoxine.
Natural extracts
• Chamomile and quinone (15 g/Kg), 87.7%
survival.
• Pine pollen (16 mg/Kg), 77.3% survival
(Phelps y Popma, 2000; Hurtado, 2005; Enciso-Lizarraga, 2008; Diaz-Neto et al., 2017;
Nian et al., 2017; Delgadillo, 2020)
4
Bioactive compounds
Sulfated
polysaccharides,
Phenolic compounds,
flavonoids and
carotenoids
•
•
•
•
Antifungal
Antibacterial
Antioxidants
Inmunoestimulants
(Aguilar-Briseño et al., 2015; Osuna et al., 2016; Murthy et al., 2017; Gomez-Zavaglia et al., 2019)
5
Reported mortality rates at different MT dosages
Species
MT
Male %
Mortality % Source
Oreochromis spirulus
70 mg/Kg
90.3%
60%
Ridha y Lone (1990)
Oreochromis spp
60 mg/Kg
100%
23%
Lopez et al. (2007)
Xiphophorus hellerii
40 mg/Kg
99%
15%
Mousavi-Sabet y Ghasemnezhad
(2013)
O. niloticus
60 mg/Kg
99%
17%
Soltan et al. (2013)
X. variatus
80 mg/Kg
96%
20%
Faghani-Langroudi et al. (2014)
6
Reported benefits of macroalgae administration in finfish
Fish species
Algae
Modality
Inclusion
rate
Benefits
Source
Oreochromis
niloticus
Ascophyllum
nodosum
Dry meal
20 g/Kg
41% weight gain
Costa et al.
(2013)
Dicentrarchus
labrax
Fucus spp.
Dry meal
7.5%
20% antioxidant activity
increment
Peixoto et al.
(2016)
Oreochromis
niloticus
Sargassum
cristaefolium
HCl extract
and ETOH
precipitate
Higher phagocitic activity,
leucocytes and plasma total
protein
Inansetyo et al.
(2016)
Oreochromis
niloticus exposed
to methomyle
(4.015 ppm)
Sargassum
spp.
Methanol
extract
0.4 mg/Kg
200 ppm
Augmented eritrocyte count Kilawati e
by 317%.
Islamy (2019)
2,2-diphenyl-1-picrylhydrazyl (DPPH) and ABTS reduction REPORTED BY SEVERAL AUTHORS
7
PLANTEAMIENTO DEL PROBLEMA
Oreochromis niloticus
Padina durvillaei
17α-methyl-testosterone
8
HYPOTHESIS
The inclusion of an aqueous extract of P. durvillaei, reduces O. niloticus
mortality during sex reversal with 17α-MT, and increases stress
tolerance in handling after this process.
General Objetive:
• Evaluate the potential positive effects of an aqueous extract of P. durvillaei
on survival and growth parameters of Nile tilapia fry during 17α-MT sex
reversal.
9
METODOLOGÍA
Macroalgae recolection.
Figure 1. Algae sampling area
Algae extract procesing: Patent WO 2014/167162 Al (SineiroTorres et al., 2014).
10
Tabla. Experimental treatments
Diets
17α-MT conc.
Macroalgae
inclusion rate
C-
0 mg/Kg
0%
C+
60 mg/Kg
0%
E1
60 mg/Kg
0.5%
E2
60 mg/Kg
1.5%
E3
60 mg/Kg
3%
• Proximal analysis AOAC (2000)
• Bioactive compounds: spectophotometry tecniques Técnicas Benítez et al., 2020).
• Antioxidant activity in each diet: radical DPPH (Müller et al., 2011); and radical ABTS (Przygodzka et al.,
2014; modified por Osuna, 2016)
11
28 days feeding
•
•
•
•
•
Triplicate
12 per liter, daily 70% water exchange
9 feedings per day
Temperature (29.2°C)
Natural photoperiod in plastic greenhouse
Stress test por masculinization
• Packing and transport simulation for 6,
12 and 24 h.
• Survival and growth parameters.
• Sexing using aceto-carmine squash
method (Guerrero y Shelton, 1974)
12
Sexing using aceto-carmine squash method
(Guerrero y Shelton, 1974) (45-55 d post hatching)
a)male
b) female
RESULTS Y DISCUSION
Algal Extract
4.15% OUTPUT (2.5%-6%, Sineiro-Torres et al., 2014).
•
•
•
•
•
•
•
•
•
Soluble protein
Total Carbohydrates
Uronic acids
Sulfates
Phenolic compounds
Flavonoids
Clorophyll a
Clorophyll b
Carotenoids
1.07 ± 0.72%
17.41 ± 2.51%
2.29 ± 1.13%
22.69 ± 3.08%
259.74 ± 1.11 µg EAG/mg
47.37 ± 4.76 µg EQ/mg
2.11 ± 0.05 µg/mg
1.18 ± 0.01 µg/mg
2.86 ± 0.12 µg/mg
14
Table. Bioactive compounds per diet
Flavonoids
totales (µg
EQ/mg dry
matter)
Diet
Uronic acids
(%)
Sulfates (%)
Phenolics (µg
EAG/mg dry
matter)
C-
1.45 ± 0.15 ab
1.57 ± 1.03 b
50.24 ± 6.56 b
4.91 ± 0.092
a
C+
1.16 ± 0.46 b
1.51 ± 0.71 b
50.17 ± 7.17 b
E1
1.82 ± 0.06 a
1.58 ± 0.73 b
66.91 ± 7.43 ab 4.55 ± 1.53 a
0.2 ± 0.01 c
0.42 ± 0.03 c
0.02 ± 0.005 b
E2
1.78 ± 0.09 a
2.53 ± 0.51 b
68.91 ± 5.74 ab 4.52 ± 1.85 a
0.27 ± 0.02 b
0.56 ± 0.03 b
0.09 ± 0.02 a
E3
1.98 ± 0.11 a
5.47 ± 1.08 a
0.44 ± 0.02 a
0.87 ± 0.02 a
0.018 ± 0.016 b
86.02 ±6.11 a
Clorophyll a
(µg /mg dry
matter)
Clorophyll b
(µg /mg dry
matter)
Carotenoides
(µg /mg dry
matter)
0.32 ± 0.03 b
0.78 ± 0.06 a
0.04 ± 0.01 b
4.55 ± 0.19 a 0.24 ± 0.02 bc 0.53 ± 0.05 bc
0.04 ± 0.01 b
5.87 ± 1.83 a
IC50 carotenoides de 457 µg /mg
(Alcalá-Hernandez et al., 2017)
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Table. Growth parameters
Diets
Final ind
weigth
Final total
lenght
SGR
K
FCR
C-
0.63 ± 0.03 a
3.34 ± 0.05 a
15.02 ± 0.20 a
1.69 ± 0.02 a
0.99 ± 0.05 a
C+
0.53 ± 0.03 a
3.24 ± 0.1 a
14.59 ± 0.25 a
1.57 ± 0.06 ab
1.18 ± 0.08 a
E1
0.57 ± 0.07 a
3.29 ± 0.12 a
14.82 ± 0.48 a
1.59 ± 0.02 ab
1.11 ± 0.15 a
E2
0.56 ± 0.03 a
3.34 ± 0.07 a
14.78 ± 0.22 a
1.51 ± 0.02 b
1.12 ± 0.07 a
E3
0.57 ± 0.04 a
3.3 ±0.1 a
14.81 ± 0.31 a
1.58 ± 0.02 b
1.11 ± 0.1 a
16
0
0
0
0
4
13
28
37
42
Frecuencia (no. de peces)
1
0
6
19
0
0
0
0
3
6
34
34
Frecuencia (no. de peces)
0
2
4
12
0
1
0
0
0
8
28
0.10.20.30.40.50.60.70.80.9 1 1.11.21.3
Peso (g)
0.10.20.30.40.50.60.70.80.9 1 1.11.21.3
Peso (g)
55
e)
40
Frecuencia (no. de peces)
0
0
0
26
d)
c)
0.10.20.30.40.50.60.70.80.9 1 1.11.21.3
Peso (g)
47
0.10.20.30.40.50.60.70.80.9 1 1.11.21.3
Peso (g)
b)
Frecuencia (no. de peces)
0
3
6
23
50
47
14
3
4
0
0
0
0
Frecuencia (no. de peces)
1
0
3
4
36
43
34
16
9
2
1
1
0
a)
0.10.20.30.40.50.60.70.80.9 1 1.11.21.3
Peso (g)
Figura. Frecuency histogram for final individual weight
17
Tabla 8. Final survival after stress test and sex ratio por treatment
Diet
Survival rate
% Male ratio
C-
97.73 ± 3.46 a
67% c
C+
97.98 ± 1.58 a
100% a
E1
99.27 ± 0.82 a
89% b
E2
99.12 ± 0.28 a
89% b
E3
96.8 ± 3.39 a
87% b
18
Paradoxical feminization
Androgen aromatization
(Solar et al., 1984; Beardmore et al., 2001; Fenske y Segner, 2004)
19
Receptor biding competition
17α-MT
(Vega-Galarza, 2018)
(Eng et al., 2001; Rodriguez-Montes de Oca, 2005; Omeje et al., 2020).
20
Competencia
Compuestos fitoquímicos
17α-metil-testosterona
•
•
•
•
Fenoles (Controles < Dietas suplementadas)
Flavonoides
Taninos
Saponinas
21
Conclusions
●Modification of polysacchares extraction to
improve expected results.
●What specific extracted fraction or compound has
an endocrine disrupture protective effect?.
22
¡GRACIAS!
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