SciELO - Scientific Electronic Library Online

 
vol.45 número5Abundance estimate of the Amazon River dolphin (Inia geoffrensis) and the tucuxi (Sotalia fluviatilis) in southern Ucayali, PeruAssessment of some nongenetic factors that affect egg mass weight of channel catfish, Ictalurus punctatus índice de autoresíndice de materiabúsqueda de artículos
Home Pagelista alfabética de revistas  

Servicios Personalizados

Revista

Articulo

Indicadores

Links relacionados

  • En proceso de indezaciónCitado por Google
  • No hay articulos similaresSimilares en SciELO
  • En proceso de indezaciónSimilares en Google

Compartir


Latin american journal of aquatic research

versión On-line ISSN 0718-560X

Lat. Am. J. Aquat. Res. vol.45 no.5 Valparaíso nov. 2017

http://dx.doi.org/10.3856/vol45-issue5-fulltext-12 

Research Article

Antioxidant activity and apparent digestibility of amino acids of three macroalgae meals in the diets of Pacific white shrimp (Litopenaeus vannamei)

Ricardo Ernesto Anaya-Rosas1 

Mario Nieves-Soto1 

Martha Elisa Rivas-Vega2 

Anselmo Miranda-Baeza2 

Pablo Piña-Valdez1 

1Universidad Autónoma de Sinaloa, Mazatlán, Sinaloa, México

2Universidad Estatal de Sonora, Carretera a Huatabampo, Sonora, México

ABSTRACT

The antioxidant activity of meals, growth performance, dry matter, crude protein and amino acid apparent digestibility coefficients (ADC) were determined for three macroalgae in Pacific white shrimp juveniles: Gracilaria vermiculophylla, Dictyota dichotoma and Ulva lactuca. For the digestibility determination, test diets included 15% of the test ingredients and 85% of a control diet supplemented with 1% chromic oxide. The amino acid content in the ingredients, diets and feces were analyzed using HPLC. In general, nutrient digestibility values were far higher in the diets with macroalgae meals than in the control diet. In conclusion, diets with the macroalgae U. lactuca and G. vermiculophylla in particular showed high antioxidant activity, high amino acid digestibility and improved Litopenaeus vannamei growth.

Keywords: shrimp; macroalgae; digestibility; amino acid; antioxidant

INTRODUCTION

Macroalgae represent a source of natural bioactive compounds (gallic acid, catechin, epicatechin and others) (López et al., 2011). It has been discovered that the antioxidant capacity of various species of brown algae, which are associated with high levels of phenolic compounds, contributes to the ability to neutralize the effects of oxidative stress that is associated with ageing in living organisms (Cano-Mallo, 2008).

Macroalgae have been considered a source of nutrients in the diet of aquatic organisms. Some studies have reported the growth levels of shrimp fed with diets containing macroalgae meal. Rodríguez-González et al. (2014) evaluated Ulva lactuca and Gracilaria parvispora meals, and they concluded that both macroalgae may be used as a source of protein in balanced diets for shrimp. However, except for a study that evaluated the crude nutrient digestibility of the macroalgae Macrocystis pyrifera and Sargassum sp. in diets for Litopenaeus vannamei (Gutiérrez-Leyva, 2006), no work has been carried out specifically to evaluate the amino acid digestibility of macroalgae in shrimp diets.

The genus of macroalgae most commonly studied in the field of aquaculture nutrition, effluent bioremediation and shrimp immunostimulation are Gracilaria sp. and Ulva sp. In this study, three macroalgae, Gracilaria vermiculophylla, Dictyota dichotoma and Ulva lactuca, were selected to evaluate the antioxidant activity and their effects on shrimp growth and amino acid digestibility.

MATERIALS AND METHODS

Macroalgae meals

Macroalgae (G. vermiculophylla, D. dichotoma and U. lactuca) were collected from Agiabampo Bay, Sonora, Mexico (26°22′31″N, 109°13′37″W), dried in a convection oven (Shell Lab CE3F) at 60°C for 24 h, milled in a pulveriser Pulvex 200, sifted through a 250 μm mesh strainer, and stored at 4°C until its use.

Analysis of macroalgae meals

The moisture, ash, crude fiber, crude protein using Kjeldahl method and crude fat for Soxhlet method according to Association of Official Analytical Chemist (AOAC, 2002). The Nitrogen Free Extract (NFE) was calculated by the difference, using the following equation: NFE = (100 - % crude protein - % crude fat - % crude fiber - % ash). Methanolic extracts of macroalgae were prepared according to Robles-Sánchez et al. (2011). The extraction was carried out for 12 h using 99% methanol (1:12, macroalgae meal:methanol). Mixture was filtered and the supernatant was used for total phenols, DPPH and TEAC assays.

Phenols were measured spectrophotometrically using the Folin-Ciocalteu reagent, with gallic acid as the standard. Briefly, 50 μL of macroalgae extract were added to 3 mL of deionized water plus 250 μL of Folin-Ciocalteu reagent (diluted 2-fold using water as diluent before use). After 5 min, 250 μL of a 7% Na2CO3 solution were added. The mixture was made up to 5 mL with deionized water and incubated for 90 min at room temperature. The absorbance was measured at 750 nm, and the results were reported as mg of gallic acid equivalents (GAE) per 100 g of meal (Robles-Sánchez et al., 2011).

Free radical scavenging activity of macroalgae was evaluated spectrophotometrically against the absorbance of the indicator 2, 2-diphenyl-1-picrylhydrazyl (DPPH) solution. Macroalgae extracts were diluted to a final concentration of 0.06 mg mL−1, and 3.9 mL of a methanolic solution of DPPH (2.5 mg of DPPH.100 mL−1 MeOH) were mixed with 0.1 mL of each sample and shaken vigorously. The tubes were allowed to stand at 27°C for 20 min. The control was prepared as above without any extract, and MeOH was used for the baseline correction. Changes in the absorbance of the samples were measured at 515 nm. Radical scavenging activity (RSA) was expressed as mg Trolox (6- hydroxy-2, 5, 7, 8-tetramethylchroman-2-carboxylic acid) equivalents/g dry macroalgae meal (Robles-Sánchez et al., 2011).

The hydrophilic (TEAC) assay was performed using ABTS (2, 2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid)). This stable radical cation was performed by mixing 10 mL of a 7 mmol L−1 ABTS solution with 10 mL of a 2.45 mmol L−1 K2S2O8 solution. After 24 h at room temperature in the darkness, the ABTS stock solution was ready to use. An ABTS working solution was prepared daily by diluting the ABTS stock solution with a phosphate buffer (75 mmol L−1, pH 7.4) to an absorbance of 0.70 ± 0.05 at 734 nm. Twenty microliters of extract were mixed with 200 mL of an ABTS working solution on a 96-well microplate. Trolox solutions (12.5-250 mmol L−1) were used for constructing a regression line, water acted as blank, and the antioxidant capacities of the samples were expressed as mg Trolox equivalents g−1 dry macroalgae meal (Re et al., 1999, Müller et al., 2010).

Growth trial

Diet formulation and processing

Three experimental diets were elaborated, each with the inclusion of 15% of macroalgae meal and a control diet without macroalgae meal. Experimental diets were isoproteic and isocaloric, formulated with the Nutrion 5 Pro software. Ingredients such as fishmeal, soybean meal and whole-wheat flour were milled with a Cyclotec mill and strained at 250 μ. Subsequently they were mixed with an emulsion of soy lecithin and fish oil, enriched with a premix of vitamins and minerals and distilled water was added at 60°C to form a paste. This was passed through a conventional meat mill, using a 3 mm diameter die with which the pellets were formed.

Experimental diets were dried in an air convection oven at 45°C for 24 h and the product was used to feed the shrimp in culture. Table 1 shows the ingredient composition of the diets that were used in this experiment.

Table 1 Ingredient composition (g kg–1 diet) of the diets used to determine growth in L. vannamei juveniles. 

Ingredient (%) Control G. vermiculophylla D. dichotoma U. lactuca
Fishmeal (sardine)1 150.00 150.00 150.00 150.00
Wheat flour2 515.50 376.40 370.00 376.40
Soybean meal2 260.90 230.00 256.4 250.00
Macroalgae meal3 0.00 150.00 150.00 150.00
Sodium alginate4 25.00 25.00 25.00 25.00
Vitamin premix5 8.00 8.00 8.00 8.00
Mineral premix6 5.00 5.00 5.00 5.00
Choline chloride 62 %7 2.00 2.00 2.00 2.00
Vitamin C8 1.00 1.00 1.00 1.00
Dibasic sodium phosphate9 5.00 5.00 5.00 5.00
Fish oil (sardine)1 17.50 17.50 17.50 17.50
Soybean lecithin 110 10.00 10.00 10.00 10.00
BHT11 0.10 0.10 0.10 0.10

1Industrias Barda, Yavaros, Sonora, Mexico.

2Alimentos Colpac S.A. de C.V.

3Prepared in the laboratory from macroalgae collected at Agiabampo Bay, Sonora, México.

4Química Meyer Cat. Num. 6780. México, D.F.

5Composition of the vitamin premix (g kg−1 premix): Vit. A (20,000 UI g−1) 5.6, D3 (850,000 UI g−1) 0.001, DL-α-tocopheryl acetate (250 UI g−1) 8.9, Menadione 2.2, Thiamin-HCl 0.6, Riboflavin 3.3, Pyridoxine-HCl 1.1, DL-Ca-Pantothenate 5.6, nicotinic acid 5.6, Biotin 0.1, Inositol 5.6, B12 0.002, folic acid 0.2, alpha-cellulose 961.4.

6Composition of the mineral premix (g 100 g−1 premix): CoCl2 0.004, CuSO4.5H2O 0.25, FeSO4.7H2O 4, MgSO4.7H2O 28.398, MnSO4.H2O 0.65, KI 0.067, Na2SeO3 0.01, ZnSO4.7H2O 13.193, alfa-celulose 53.428.

7SIGMA Cat. Num. C1879. SIGMA-ALDRICH chemical company, St. Louis, MO, USA.

8Stay C 35% active agent. Roche, México, D.F.

9SIGMA Cat No. S-0876. SIGMA-ALDRICH Chemical Company, St. Louis, MO, USA

10ODONAJI, distribuidora de alimentos naturales y nutricionales (distributor of natural and nutritional food) S.A. de C.V. México, D.F.

11Butylated hydroxytoluene, ICN Cat. No.101162. Aurora, Ohio, USA.

Selection of shrimp juvenile

Fifteen hundred juvenile of L. vannamei were obtained from a hatchery in Sonora south, a temperature of 20°C was maintained during the transfer to the bioassay laboratory at the Universidad Estatal de Sonora.

Juvenile white shrimp (L. vannamei), with a mean weight of 0.054 ± 0.012 g, were stocked in 60 L rectangular tanks at a density of 10 shrimp/tank. Three replicate tanks were randomly assigned for each diet. Shrimps were maintained in filtered seawater at 28 ± 1°C, 35 ± 0.04 of salinity, and 5.0 ± 0.11 mg L−1 dissolved oxygen. The shrimp were fed ad libitum three times daily (08:00, 13:00 and 18:00 h). At the beginning of the trial, the diet was introduced at a 10% rate of the biomass in each tank. During the following days, the amount was corrected based on the residual food. The shrimp were weighed every 15 days for a period of 45 days. The following response variables were determined for each experimental tank: mean body weight; growth rate expressed as percentage weight gain (%WG) = 100 × (initial weight-final weight)/(initial weight); feed conversion ratio (FCR) = (total feed intake)/(weight gain); and protein efficiency ratio (PER) = (weight gain)/(protein intake); and survival (%S) =100 × (final count)/(initial count).

Apparent digestibility trial

Diet formulation and processing

Four diets for the digestibility bioassay were formulated and elaborated with the same composition and methodology as those developed for the growth bioassay. In contrast, these diets were added 1% chromium oxide (Cr2O3) as an inert marker. Table 2 shows the ingredient composition of the diets that were used in this experiment.

Table 2 Ingredient composition (g kg−1 diet) of diets used to determine digestibility in L. vannamei juveniles. 

Ingredient (%) Control G. vermiculophylla D. dichotoma U. lactuca
Fishmeal (sardine)1 150.00 148.5 148.5 148.5
Wheat flour2 480.00 376.20 366.40 372.60
Soybean meal2 280.00 247.50 258.40 247.50
Macroalgae meal3 0.00 148.50 148.50 148.50
Sodium alginate4 24.70 24.80 24.80 24.80
Vitamin premix5 7.80 7.80 7.80 7.80
Mineral premix6 5.00 5.00 5.00 5.00
Choline chloride 62%7 1.90 1.90 1.90 1.90
Vitamin C8 1.00 1.00 1.00 1.00
Dibasic sodium phosphate9 5.00 5.00 5.00 5.00
Fish oil (sardine)1 17.40 17.40 17.40 17.40
Soybean lecithin 110 9.9 9.9 9.9 9.9
BHT11 0.10 0.10 0.10 0.10
Chromic oxide12 10.00 10.00 10.00 10.00

1Industrias Barda, Yavaros, Sonora, Mexico.

2Alimentos Colpac S.A. de C.V.

3Prepared in the laboratory from macroalgae collected at Agiabampo Bay, Sonora, México.

4Química Meyer Cat. Num. 6780. México, DF,

5Composition of the vitamin premix (g kg-1 premix): Vit. A (20,000 UI g-1) 5.6, D3 (850,000 UI g-1) 0.001, DL-α-tocopheryl acetate (250 UI g-1) 8.9, Menadione 2.2, Thiamin-HCl 0.6, Riboflavin 3.3, Pyridoxine-HCl 1.1, DL-Ca-Pantothenate 5.6, nicotinic acid 5.6, Biotin 0.1, Inositol 5.6, B12 0.002, folic acid 0.2, alpha-celulose 961.4,

6Composition of the mineral premix (g 100 g-1 premix): CoCl2 0.004, CuSO4.5H2O 0.25, FeSO4.7H2O 4, MgSO4.7H2O 28.398, MnSO4.H2O 0.65, KI 0.067, Na2SeO3 0.01, ZnSO4.7H2O 13.193, alfa-celulose 53.428,

7SIGMA Cat. Num. C1879. SIGMA-ALDRICH Chemical Company, St. Louis, MO, USA,

8Stay C 35% active agent. Roche, México, DF,

9SIGMA Cat No. S-0876. SIGMA-ALDRICH Chemical Company, St. Louis, MO, USA,

10ODONAJI, distribuidora de alimentos naturales y nutricionales (distributor of natural and nutritional food) S.A. de C.V. México, DF,

11Butylated hydroxytoluene, ICN Cat. No.101162. Aurora, Ohio, USA,

12Aldrich Cat. No. 20,216-9. SIGMA-ALDRICH Chemical Company, St. Louis, MO, USA.

Selection of shrimp juvenile

Juvenile white shrimp (L. vannamei), with a mean weight of 8.2 ± 1.2 g, were obtained from a shrimp farm located in Sonora south and stocked in 60 L rectangular tanks at a density of 10 shrimp/tank. Three replicate tanks were randomly assigned for each diet. The shrimp were maintained for acclimation in filtered seawater at 29 ± 1°C, 35 of salinity and 6 mg L−1 dissolved oxygen. The shrimp were fed with the experimental diets ad libitum three times daily for 7 days before feces collection began. One hour after each feeding, fecal strands were siphoned, gently rinsed with distilled water, and frozen at −60°C. At the end of the trial, the feces collected from each tank were pooled and freeze- dried. Diets and fecal samples were analyzed for chromic oxide (Olvera-Novoa, 1994), crude protein (AOAC, 2002) and amino acids (Umagat et al., 1982; Jones & Gilligan, 1983). The apparent digestibility coefficients (ADC) for nutrients of test ingredient were determined according to Bureau & Hua (2006) using the following equation:

ADE test ingredient=ADC test diet+[(ADC test dietADC ref.diet)×(0.85*D ref.0.15*D ing.)]

where D ref. = % nutrient of reference diet; D ing. = % nutrient of test ingredient.

Amino acid composition

Amino acids were determined after hydrolysis of the macroalgae, fecal and diet samples were placed in 6 N HCl for 6 h at 150°C. Samples were then dried using a rotary evaporator (Yamato, RE301) and re-suspended in 2 mL of 6N HCl. The total amino acids were determined via HPLC (HITACHI L-8900 amino acid analyser) using an ion exchange column (HITACHI # 2622SC-PH) at a flow rate of 1 mL min–1 with a fluorescence detector (Umagat et al., 1982; Jones & Gilligan, 1983).

Statistical analysis

A one-way ANOVA was applied to determine significant differences among treatments. Tukey's multiple range test was used to identify differences among means. All statistical analyses were performed at the 0.05 significance level using Statistica™ 7.0 (StatSoft Inc., Tulsa, OK, USA).

RESULTS

Table 3 shows the results of the total phenol content of macroalgae meals. Concentration of total phenols in G. vermiculophylla was significantly higher than in U. lactuca and D. dichotoma (14.78, 11.59 and 11.68 mg GA100 g−1 dry wt, respectively) and exhibited the greatest antioxidant activity expressed as DPPH radical scavenging activities.

Table 3 Antioxidant activity of macroalgae extracts. Mean ± SD of three replicates. Values in the same column with different superscripts are significantly different (P < 0.05). 

Macroalgae Total phenolic content (mg GA equivalents g−1 dry macroalgae meal) DPPH radical scavenging activities (mg Trolox equivalents g−1 dry macroalgae meal) ABTS (mg Trolox equivalents g−1 dry macroalgae meal)
Gracilaria vermiculophylla 14.78a ± 1.47 0.21a ± 0.01 40.0a ± 0.6
Dictyota dichotoma 11.59b ± 1.16 0.13b ± 0.01 28.4b ± 2.5
Ulva lactuca 11.68b ± 2.19 0.14b ± 0.02 37.1a ± 4.7

In the growth trial, significant differences were found between the means of the treatments, where the U. lactuca fed shrimp obtained a final weight higher than the rest of the treatments. The shrimp cultivated with the control diet, to which no macroalgae were added, grew below the rest of the treatments. On the other hand, in the survival and in the feed conversion factor, no significant differences (P > 0.05) were found between the treatments (Table 4).

Table 4 Growth performance of juvenile L. vannamei fed on different macroalgae meals. Mean ± SD of three replicates. Numbers in the same column with different superscripts are significantly different (P < 0.05). 

Diet Final weight (g) Survival (%) FCR1 RGR2 (%)
Control 2.41c ± 0.28 90c ± 0.03 2.57a ± 0.42 4.319a ± 569
G. vermiculophylla 3.37ab ± 0.28 97b ± 0.01 2.40a ± 0.19 5.469a ± 398
D. dichotoma 3.06bc ± 1.04 100a ± 0.00 2.77a ± 0.54 5.698a ± 1,89
U. lactuca 3.71a ± 0.25 100a ± 0.00 2.54a ± 0.16 6.804a ± 356

1Factor conversion ratio,

2Relative growth ratio.

The ADC of dry matter and crude protein for the experimental diet of 150 g kg−1 of macroalgae meal are shown in Table 5. The apparent digestibility of the diet changed from 41.6 to 72.1%, with G. vermiculophylla recording the best ADC for dry matter followed by U. lactuca and D. dichotoma. The ADC of crude protein was similar for diets containing macroalgae meal, which were higher than the results recorded for the control diet.

Table 5 Apparent digestibility coefficients for dry matter and crude protein of diets containing macroalgae meals. Mean ± SD of three replicates. Values in the same column with different superscripts are significantly different (P < 0.05). 

Diet Dry matter (%) Crude protein (%)
Control 41.60c ± 0.45 56.44b ± 0.34
G. vermiculophylla 72.06a ± 3.68 81.07a ± 2.91
D. dichotoma 60.83b ± 0.82 78.80a ± 2.47
U. lactuca 61.72b ± 1.26 82.19a ± 1.43

Table 6 shows the amino acid ADC of the diets containing macroalgae meal. The highest ADC was obtained by the diet formulated with G. vermicullophylla meal (92.8%), and the lowest was obtained by the control diet (67.3%), to which macroalgae meal was not added. Alanin (54.9%) was the least digestible amino acid in all samples, while arginine and lysine (87.4 and 85.7%, respectively) were generally the most easily digested.

Table 6 Apparent digestibility coefficients for amino acids of diets containing macroalgae meals Mean ± SD of three replicates. Values in the same row with different superscripts are significantly different (P < 0.05). 

Control Gracilaria vermiculophylla Dictyota dichotoma Ulva lactuca
Essential amino acids (EAA)
Arginine 76.82bc ± 3.31 95.28a ± 0.17 88.91b ± 0.69 86.92b ± 0.68
Histidine 67.89c ± 5.34 93.60a ± 1.22 83.68b ± 0.45 81.49b ± 1.73
Isoleucine 68.38c ± 1.01 93.33a ± 1.26 84.43b ± 0.33 78.36b ± 1.87
Leucine 71.85c ± 2.67 93.79a ± 1.17 85.28b ± 0.26 82.33b ± 1.76
Lysine 73.23c ± 0.16 94.91a ± 2.87 87.28b ± 3.75 85.31b ± 1.11
Methionine 59.95c ± 2.50 92.61a ± 1.37 81.71b ± 0.78 76.19b ± 1.10
Phenylalanine 69.36c ± 3.30 93.19a ± 1.50 84.13b ± 0.74 80.54b ± 1.62
Valine 70.04c ± 0.82 93.19a ± 1.45 85.11b ± 0.67 80.84b ± 2.96
Non-essential amino acids (NEAA)
Alanine 54.87c ± 9.94 90.56a ± 1.98 80.59b ± 7.34 79.75b ± 4.27
Glutamic acid 68.68c ± 2.21 88.41b ± 0.00 94.24a ± 5.55 87.75b ± 1.20
Glycine 56.42c ± 8.50 90.96a ± 0.47 77.21b ± 4.72 78.06b ± 2.34
Serine 70.00c ± 0.00 93.14a ± 0.17 76.02b ± 0.00 82.27b ± 2.91
Tyrosine 68.08c ± 1.22 93.25a ± 1.26 82.60b ± 1.78 80.43b ± 1.35

DISCUSSION

Antioxidant activity

Wang et al. (2009) found that brown algae generally contained higher amounts of polyphenols than red and green algae, and in our study, we did not find this trend, as the red algae G. vermiculophylla presented the highest content of phenols.

Some authors have found a positive correlation between the content of total phenols and the antioxidant activity of macroalgae (López et al., 2011; Raja et al., 2016), and this behavior was also found in our study; G. vermiculophylla presented the highest content of total phenols as well as antioxidant activity, measured as DPPH radical scavenging activities and ABTS. The high scavenging property of G. vermiculophylla may be due to hydroxyl groups that exist in the phenolic compounds (Raja et al., 2016).

In general, the use of macroalgae as an ingredient in diets for L. vannamei has not been observed to affect negatively the survival (Peñaflorida & Golez, 1996; Cruz-Suárez et al., 2000; Gutiérrez-Leyva, 2006; Da Silva & Barbosa, 2008; Peña-Rodríguez et al., 2010).

Chemical composition of macroalgae meal and shrimp growth

The proximal chemical composition of the macroalgae studied is very similar to that reported by Cruz-Suárez et al. (2010) for U. clathrata, as well as for U. pertusa and U. intestinalis (Benjama & Masniyom, 2011). The proximal analysis of the algae meals studied shows that they have nutritional characteristics superior to those reported by Gutiérrez-Leyva (2006) for Sargassum sp. and kelp, where meals of these algae were used as diet ingredients for L. vannamei with levels of inclusion of 1, 4, 7 and 10% for both algae.

The final weight increased significantly (P < 0.05) in shrimp fed with G. vermiculophylla and U. lactuca.Gutiérrez-Leyva (2006) found similar results with M. pyirifera and Sargassum sp., where the final weight of shrimp was higher than that of shrimp fed with the control diet. Peña-Rodríguez et al. (2010) shows the benefits obtained by the shrimp when fed with macroalgae, especially those of the genus Ulva. Diets prepared with inclusion rates of less than 5% of macroalgae meals give favorable results with respect to untreated organisms, mainly in terms of weight gain and shrimp coloration after a cooking process.

Macroalgae have been associated with the presence of bioactive compounds with antioxidant, antimicrobial and anti-inflammatory effects (Banerjee et al., 2009; Plaza et al., 2010; Raja et al., 2016), an association that can also be made from the results of this study, considering that these macroalgae contained higher contents of total phenols and higher antioxidant activity.

Macroalgae of the genus Gracilaria are an important source of phytocolloid (Troell et al., 2003); the green macroalgae of the genus Ulva containing the polysaccharides ulvans (Sathivel et al., 2008). Algal polysaccharides have been demonstrated to play an important role as free-radical scavengers and antioxidants for the prevention of oxidative damage in living organisms (Kim et al., 2007; Wang et al., 2009; Souza et al., 2012). In the present study the macroalgae were supplied complete to organisms of L. vannamei and it is possibly that this components had an important effect in the growth and survival response.

The feed conversion and the relative growth ratios were unaffected, with Cruz-Suárez et al. (2000) finding similar results in their evaluation of M. pyrifera. While macroalgae represent an important source of protein and energy, information about the availability of nutrients for aquatic organisms is scarce (Cruz-Suárez et al., 2008). Most researches focus on the use of vegetable matter as a substitute for fishmeal.

The protein and energy composition of macroalgae is a viable source to be used as an ingredient in shrimp diets. The few studies on its use make it necessary to know more about the bioavailability of these nutrients for aquatic organisms (Cruz-Suárez et al., 2008). Most of the research is focused on the use of terrestrial vegetable sources in the substitution of fishmeal.

The relationship between weight gain and survival (Table 4) with ADC of dry matter, protein (Table 5) and amino acids (Table 6) in diets, is something to be emphasized in this study, due to the shrimp that were treated with diet containing macroalgae meal as an ingredient, obtained better results than the untreated organisms. This suggests that the nutrient content of macroalgae (Cruz-Suárez et al., 2010) and the presence of antioxidant compounds and other bioactive compounds (Esquer-Miranda et al., 2016) improve the health and well-being of shrimp, which allows efficient nutrition, weight gain and survival greater than the control organisms.

Digestibility protein and amino acids

Several aspects determine the nutritional quality of an ingredient as a nutrient source specifically: its palatability, level of the nutrient, anti-nutritional factors and digestibility of the nutrients (Divakaran et al., 2000). Studies conducted to ascertain the nutrients digestibility coefficients are most commonly conducted by feeding a diet containing fixed levels of a nutrient source, the inclusion commonly used of test ingredients in experimental diets is 30%, however, some authors had used different levels of inclusion: 88% (Akiyama et al., 1989), 50% (Kumaraguru et al., 2007) and 15% (Divakaran et al., 2000). Found that the dietary soybean meal level did not consistently affect the ADC values calculated for this ingredient, in Litopenaeus vannamei diets. In the present study, the level of macroalgae meals is 15% and is assumed that the estimation of digestibility is not affected by the inclusion.

Non-significant differences among treatments were found for the amino acid ADC of the macroalgae meal. Terrazas-Fierro et al. (2010) evaluated marine foodstuffs and found that these ingredients are good sources of available protein and amino acids for juvenile white leg shrimp. Despite being a vegetable protein source, macroalgae presented high amino acid digestibility, in contrast with non-marine vegetable foodstuffs (Oujifard et al., 2012).

The in vivo ADC of dry matter and crude protein for the experimental diets added with 15% of meal from each studied macroalgae are shown in Table 5. The apparent digestibility of diets varied from 41.60 to 82.19%. It was found that the diets with macroalgae were significantly (P < 0.05) more digestible than the control diet, being concurrent with a higher growth of the cultivated organisms comparable to that reported by Oujifard et al. (2012), in diets used for L. vannamei by partially replacing fishmeal with rice protein concentrate. The results for ADC of dry matter and protein are similar to those reported by Oujifard et al. (2012), with values for dry matter between 60 and 85% between the experiments.

Table 7 Apparent digestibility coefficients for essential and non-essential amino acids in macroalgae meals. Mean ± SD of three replicates. Only non-significant differences among treatments were found (P > 0.05). 

Gracilaria vermiculophylla Dictyota dichotoma Ulva lactuca
Essential amino acids (EAA)
Arginine 68.6 ± 7.7 71.9 ± 9.1 66.1 ± 2.2
Histidine 88.5 ± 5.8 85.9 ± 1.5 71.8 ± 6.0
Isoleucine 75.4 ± 4.0 63.3 ± 2.3 68.9 ± 6.5
Leucine 84.2 ± 2.1 88.5 ± 0.9 70.6 ± 2.8
Lysine 82.8 ± 1.9 80.3 ± 4.4 69.3 ± 3.5
Methionine 74.7 ± 2.5 76.5 ± 1.2 82.3 ± 3.6
Phenylalanine 76.0 ± 2.1 92.0 ± 2.4 72.3 ± 3.4
Valine 82.4 ± 3.9 89.1 ± 2.0 65.2 ± 2.2
Non-essential amino acids (NEAA)
Alanine 69.6 ± 2.6 68.8 ± 2.0 81.7 ± 1.1
Glutamic acid 83.6 ± 2.7 78.5 ± 1.4 79.1 ± 2.9
Glycine 84.6 ± 2.7 77.6. ± 2.0 76.6 ± 3.5
Serine 66.4 ± 0.9 71.6 ± 1.8 67.7 ± 1.5
Tyrosine 89.5 ± 1.6 92.1 ± 2.8 74.8 ± 1.3

Similar studies have been performed on shrimp culture using M. pyrifera, Ascophylum nodosum and Ulva Clathrata as ingredients in diets, using 3.33% inclusion (Cruz-Suárez et al., 2008), studying the zootechnical parameters of the culture. Wong et al. (2001) studied the in vitro protein digestibility and amino acid profile of three species of macroalgae (Hypnea charoides, Hypnea japonica and Ulva lactuca) where they found that the digestibility of the U. lactuca protein (85.7%) was very similar to what was found in this study (82.2%). In relation to the reports made by Ramos-Díaz et al. (2001), for a dry matter and protein CDA of 78.6% and 94%, respectively, for the diet formulated with the macroalga Lessonia sp., with a percentage of inclusion of 30.9%, in the present study, lower results were obtained, of 64.8% and 80.7%, respectively, when using an inclusion level of 15%.

The results obtained in the amino acid digestibility of the tested diets are shown in Table 6, the best CDA for amino acids, was obtained in the diet added with G. vermiculophylla meal, while the control diet had the lowest amino acid digestibility.

The amino acid ADC, reported by Nieto-López et al. (2011), was slightly lower than that found in the digestibility of the amino acids in this experiment for the diet formulated with G. vermiculophylla meal. For the diets added with D. dichotoma and U. lactuca CDA were equivalent, while the control diet of this experiment resulted in an amino acid digestibility below from that reported by the previously mentioned author. The reports by Oujifard et al. (2012), where L. vannamei was grown with balanced diets, in which fishmeal was partially replaced by a rice flour protein concentrate, show the CDA results of amino acids similar to those found in this study, (81.35 to 92.9%).

CONCLUSIONS

Macroalgae, especially U. lactuca and G. vermiculophylla, demonstrated high antioxidant activity, high amino acid digestibility and improved L. vannamei growth rates. The ADC of dry matter, proteins and amino acids found in this study show the benefit of using macroalgae as an ingredient in diet for L. vannamei, where percentages between 60 and 72% for dry matter, of 78 amounted to 82% for protein and 82 to 93% for the amino acids of the experimental diets.

REFERENCES

Akiyama, D.M., S.R. Coelho, A.L. Lawrence & E.H. Robinson. 1989. Apparent digestibility of feedstuffs by the marine shrimp Penaeus vannamei Boone. Nippon Suisan Gakkaishi, 55: 91-98. [ Links ]

Association of Official Analytical Chemist (AOAC). 2002. Official Methods of Analysis. Association of Official Analytical Chemist, Washington, DC, 1094 pp. [ Links ]

Banerjee, K., R. Ghosh, S. Homechaudhuri & A. Mitra. 2009. Biochemical composition of marine macroalgae from Gangetic Delta at the apex of Bay of Bengal. Afri. J. Basic Appl. Sci., 1: 96-104. [ Links ]

Benjama, O. & P. Masniyom. 2011. Nutritional composition and physicochemical properties of two green seaweeds (Ulva pertusa and U. intestinalis) from the Pattani Bay in Southern Thailand. Songklanakarin J. Sci. Technol., 35(5): 575-583. [ Links ]

Bureau, D.P. & K. Hua. 2006. Letter to the Editor of Aquaculture. Aquaculture, 252: 103-105. [ Links ]

Cano-Mallo, M. 2008. Bases biológicas de Ulva fasciata Delile, (Chlorophyta) para su posible explotación, al oeste de la Habana, Cuba. Ph.D. Thesis, Universidad de la Habana, La Habana, 110 pp. [ Links ]

Cruz-Suárez, L.E., D. Rique-Marie, M. Tapia-Salazar & C. Guajardo-Barbosa. 2000. Uso de la harina de kelp (Macrocystis pyrifera) en alimentos para camarón. Avances en nutrición acuícola V. Memorias del V Simposium Internacional de Nutrición Acuícola. Universidad Autónoma de Nuevo León, San Nicolás de los Garza, 12-22 Noviembre 2000, pp. 227-266. [ Links ]

Cruz-Suárez, L.E., M. Tapia-Salazar, M. Nieto-López & D. Rique-Marie. 2008. A review of effects of macroalgae in shrimp feeds and co-culture. Simposio Internacional de Nutrición Acuícola. Nuevo León, Monterrey, 24-27 Noviembre 2008, 304-333 pp. [ Links ]

Cruz-Suárez, L.E., A. León, A. Peña-Rodríguez, G. Rodríguez-Peña, B. Moll & D. Ricque-Marie. 2010. Shrimp/Ulva co-culture: a sustainable alternative to diminish the need for artificial feed and improve shrimp quality. Aquaculture, 301: 64-68. [ Links ]

Da Silva, R.L. & J.M. Barbosa. 2008. Seaweed meal as a protein source for the white shrimp Litopenaeus vannamei. J. Appl. Phycol., 21: 193-197. [ Links ]

Divakaran, S., M. Velasco, E. Beyer, I. Forster & A.G. Tacon. 2000. Soybean meal apparent digestibility for Litopenaeus vannamei, including a critique of methodology. In: L.E. Cruz- Suárez, D. Ricque-Marie, M. Tapia-Salazar, M.A. Olvera-Novoa & R. Civera-Ceveredo (eds.). Avances en nutrición acuícola. Memorias del V Simposium Internacional de Nutrición Acuícola. Universidad Autónoma de Nuevo León, San Nicolás de los Garza, 12-22 Noviembre 2000, pp. 227-266. [ Links ]

Esquer-Miranda, E., M. Nieves-Soto, M.E. Rivas-Vega, A. Miranda-Baeza & P. Piña-Valdez. 2016. Effects of methanolic macroalgae extracts from Caulerpa sertularioides and Ulva lactuca on Litopenaeus vannamei in the presence of Vibrio bacteria. Fish Shellfish Immunol., 51: 346-350. [ Links ]

Gutiérrez-Leyva, R. 2006. Uso de harinas de Macrocystis pyrifera y Sargassum sp. en alimentos balanceados para camarón Litopenaeus vannamei: efectos sobre el crecimiento y la digestibilidad in vivo. Master Thesis, Centro de Investigaciones Biológicas del Noroeste, La Paz, B.C., México, 94 pp. [ Links ]

Jones, B.B. & J.P. Gilligan. 1983. O-Ophthalaldehyde precolumn derivatization and reversed-phase high-performance liquid chromatography of polypeptide- hydrolisates and physiological fluids. J. Chromatogr., 26: 471-482. [ Links ]

Kim, S.H., D.S. Choi, Y. Athukorala, Y.J. Jeon, M. Senevirathne & C.K. Rha. 2007. Antioxidant activity of sulfated polysaccharides isolated from Sargassum fulvellum. J. Food Sci. Nutr., 12: 65-73. [ Links ]

Kumaraguru, V., T. Balasubramanian & R. Venkatesan. 2007. Apparent digestibility of differently processed grain legumes, cow pea and mung bean in black tiger shrimp, Penaeus monodon Fabricius and associated histological anomalies in hepatopancreas and midgut. Anim. Feed Sci. Technol., 132(3): 250-266. [ Links ]

López, A., M. Rico, A. Rivero & M. Suárez de Tangil. 2011. The effects of solvents on the phenolic contents and antioxidant activity of Stypocaulon scoparium algae extracts. Food Chem., 125: 1104-1109. [ Links ]

Müller, L., S. Gnoyke, A.M. Popken & V. Böhma. 2010. Antioxidant capacity and related parameters of different fruit formulations. Food Sci. Technol., 43: 992-999. [ Links ]

Nieto-López, M., M. Tapia-Salazar, D. Ricque-Marie, D. Villarreal-Cavazos, A. Lemme & L.E. Cruz-Suárez. 2011. Digestibility of different wheat products in white shrimp Litopenaeus vannamei juveniles. Aquaculture, 319: 369-376. [ Links ]

Olvera-Novoa, M.A. 1994. Cuantificación de óxido de cromo en heces y alimentos. Nutrition of fish and crustaceans. In: FAO (eds.). A laboratory manual project. FAO, Rome, Field Document 19, pp. 1-33. [ Links ]

Oujifard, A., J. Seyfabadi, A.A. Kenari & M. Rezaei. 2012. Growth and apparent digestibility of nutrients, fatty acids and amino acids in Pacific white shrimp, Litopenaeus vannamei, fed diets with rice protein concentrate as total and partial replacement of fish meal. Aquaculture, 342-343: 56-61. [ Links ]

Peñaflorida, V.D. & N.V. Golez. 1996. Use of seaweed meals from Kappaphycus alvarezii and Gracilaria heteroclada as binders in diets of juvenile shrimp Penaeus monodon. Aquaculture. 143: 393-401. [ Links ]

Peña-Rodríguez, A., A. León, B. Moll, M. Tapia-Salazar, M.G. Nieto-López, D. Villarreal-Cavazos, D. Ricque-Marie & L.E. Cruz-Suárez. 2010. Uso de Ulva clathrata en la nutrición del camarón blanco: revisión. Avances en nutrición acuícola. Memorias del X Simposio Internacional de Nutrición Acuícola. Universidad Autónoma de Nuevo León, San Nicolás de los Garza, 8-10 Noviembre 2010, Monterrey, pp. 700-712. [ Links ]

Plaza, M., S. Santoyo, L. Jaime, R.G. García-Blairsy, M. Herrero, F.J. Sennoráns & E. Ibánnez. 2010. Screening for bioactive compounds from algae. J. Pharm. Biomed. Anal., 5: 450-455. [ Links ]

Raja, R., S.K. Hemaiswarya, K. Arunkumar & I. Carvalho. 2016. Antioxidant activity and lipid profile of three seaweeds of Faro, Portugal. Braz. J. Bot., 39: 1-9. [ Links ]

Ramos-Díaz, R., I. Miranda-Valdés & C. Molina-Segovia. 2001. Intake and apparent digestibility of three marine ingredients by white shrimp Litopenaeus vannamei (Boone, 1931). Estud. Oceanol., 20: 43-50. [ Links ]

Re, R., N. Pellegrini, A. Proteggente, A. Pannala, M. Yang & C. Rice-Evans. 1999. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biol. Med., 26: 1231-1237. [ Links ]

Robles-Sánchez, M., H. Astiazarán-García, O. Martín-Belloso, S. Gorinstein, E. Alvarez-Parrilla, L.E. de la Rosa, G. Yepiz-Plascencia & G. González-Aguilar. 2011. Influence of whole and fresh-cut mango intake on plasma lipids and antioxidant capacity of healthy adults. Food Res. Int., 44: 1386-1391. [ Links ]

Rodríguez-González, H., J. Orduña-Rojas, J.P. Villalobos-Medina, M. García-Ulloa, A. Polanco-Torres, E.S. López-Álvarez, M. Montoya-Mejía & A. Hernández-Llamas. 2014. Partial inclusion of Ulva lactuca and Gracilaria parvispora meal in balanced diets for white leg shrimp (Litopenaeus vannamei). J. Appl. Phycol. 26: 2453-2459. [ Links ]

Sathivel, A., H.R.B. Raghavendran, P. Srinivasan & T. Devaki. 2008. Anti-peroxidative and anti-hyperlipidemic nature of Ulva lactuca crude polysaccharide on D-Galactosamine induced hepatitis in rats. Food Chem. Toxicol., 46: 3262-3267. [ Links ]

Souza, W.S.B., M.A. Cerqueira, A.I. Bourbon, A.C. Pinheiro, J.T. Martins, J.A. Teixeira, M.A Coimbra & A.A. Vicente. 2012. Chemical characterization and antioxidant activity of sulfated polysaccharide from the red seaweed Gracilaria birdiae. Food Hydrocolloids, 27: 287-292. [ Links ]

Terrazas-Fierro, M., R. Civera-Cerecedo, L. Ibarra-Martínez, E. Goytortúa-Bores, M. Herrera-Andrade & A. Reyes-Becerra. 2010. Apparent digestibility of dry matter, protein, and essential amino acid in marine feedstuffs for juvenile white leg shrimp Litopenaeus vannamei. Aquaculture, 308: 166-173. [ Links ]

Troell, M., C. Halling, A. Neori, T. Chopin, A.H. Buschmann & N. Kautsky. 2003. Integrated mariculture: asking the right questions. Aquaculture, 226: 69-90. [ Links ]

Umagat, H., P. Lucera & L.F. Wen. 1982. Total aminoacid analysis using pre-column fluorescence derivatizacion. J. Chromatogr., 239: 463-474. [ Links ]

Wang, T., R. Jónsdóttir & G. Ólafsdóttir. 2009. Total phenolic compounds, radical scavenging and metal chelation of extracts from Icelandic seaweeds. Food Chem., 116: 240-248. [ Links ]

Wong, K.H., C. Peter & K. Cheung. 2001. Nutritional evaluation of some subtropical red and green seaweeds Part II. In vitro protein digestibility and amino acid profiles of protein concentrates. Food Chem., 72: 11-17. [ Links ]

Received: October 31, 2016; Accepted: June 06, 2017

Corresponding author: Martha Elisa Rivas Vega. martha.rivas@ues.mx

Corresponding editor: Alvaro Bicudo

Creative Commons License This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.