Effects of garlic-supplemented diet on the growth and survival of the whiteleg shrimp (Liptopenaeus vannamei)

Phan Thi Cam Tu, Nguyen Thi Kim Lien, Pham Thi Tuyet Ngan, Truong Quoc Phu and Huynh Truong Giang

College of Aquaculture and Fisheries, Can Tho University, Vietnam
ptctu@ctu.edu.vn

Abstract

This study was conducted to investigate the effects of garlic supplementation on the growth performance and survival of the whiteleg shrimp, Liptopenaeus vannamei. Shrimp were fed with four dietary levels of garlic: 0% (control), 0.5%, 1%, and 1.5%. After the 56-day feeding trial, survival and growth parameters were evaluated. The results showed that weight gain (WG), daily weight gain (DWG) and specific growth rate (SGR) were significantly improved in shrimp fed 1% and 1.5% garlic compared to those fed the control and 0.5% garlic diet (p <0.05). The highest survival rate and biomass of shrimp were obtained in the 1.5% garlic diet, and were significantly different compared with the control and 0.5% garlic diet. However, feed conversion ratio (FCR) in the 1% and 1.5% garlic was lower than that in the control and 0.5% garlic. In conclusion, supplementation of garlic could enhance growth performance and improve survival in L. vannamei, and the supplemented garlic concentration of 1.5% was suitable for L. vannamei.

Keywords: aquaculture, herb extract, shrimp post-larvae

Introduction

The white leg shrimp, Liptopenaeus vannamei is an economically important species and is cultivated around the world, especially in Vietnam. In 2021, the white leg shrimps’ farming area was 121,000 hectares and shrimp production had reached 655, 0000 tons (Directorate of Fisheries 2022). Due to a high/growing market demand, the shrimp cultural area and scale have been continuously expanded in Vietnam. Huong et al (2021) reported that the use of antibiotics has been widely applied in aquaculture industries for growth stimulation or disease treatments. However, the frequent and widespread use of antibiotics could increase the chance of infectious disease outbreaks occurring and the deterioration of rearing water quality (Srinivasan and Ramasamy 2009). Therefore, to mitigate the negative effects of antibiotics, new approaches have been developed not only to ensure a safe environment but also to promote the growth performance of aquaculture animals. The use of herbal plants in aquaculture as feed additives has recently received more interest for commercial applications, which have been applied in different forms, either as crudes, extracts, or active compounds (Awad and Awaad 2017; Marimuthu et al 2022).

Garlic (Allium sativum family Liliaceae) is used as an herbal medicine and has been extensively cultivated on a massive scale in many countries. It contains a variety of chemical compounds that have been shown to possess beneficial effects in enhancing digestion and growth (Lee et al 2012). In aquaculture, the inclusion of garlic supplementation in the diet is one way to promote growth, immune system, appetite, and resistance of aquatic animals to stress (Metwally 2009; Lee and Gao 2012; Zare et al 2021). Many studies have been conducted to evaluate the effects of garlic in fish species; for example, Shalaby et al (2006) and Aly and Mohamed (2010) reported that dietary garlic supplementation can increase specific growth rate and final weight as well as non-specific defense system of Nile tilapia (Oreochromis niloticus). In addition, garlic provides a beneficial effect on growth performance in juvenile starlet sturgeons (Acipenser ruthenus) (Lee et al 2012), Japanese seabass (Lateolabrax japonicus) (Xu et al 2020), rainbow trout ( Oncorhynchus mykiss) (Büyükdeveci et al 2018), and Eurasian Perch (Perca fluviatilis) (Zare et al 2021). To the best of our knowledge, little information is available on the effects of garlic extract on the growth of the white leg shrimp.

The objective of this study is to investigate the effects of different levels of garlic extract supplementation on the growth performance and survival of the whiteleg shrimp. The results from this study provide useful and practical strategies that could be applied in the shrimp culture industry, which is now declining using antibiotics in aquaculture.

Materials and methods

Preparation of experimental diets

Fresh whole garlic was purchased from the local market in Can Tho city, Vietnam, which was washed with tap water, peeled, sliced, and dried in the oven at 40^oC. After drying, the garlic was then powdered using a pestle and sieved using a fine wire mesh household sifter. The garlic extract was prepared following the procedure of Ng and See (2019) with a slight modification. Briefly, 10 g of the powdered garlic were extracted with 100 ml of distilled water in a water bath at 80^oC for 20 min. The resulting extract was centrifuged at 5000 rpm for 15 min to remove impurities. The extracts were stored at -4℃ in dark until using.

A commercial basal diet (Grobest Landfound Co. Ltd., Vietnam) containing 40% crude protein, 11% moisture, 4% lipid, and 5% fiber was used as a control diet. Three basal diets were supplemented with different levels of garlic extract (0.5%, 1.0%, and 1.5%). To prepare the diets, the garlic extract was blended thoroughly with an additional 1% binder, and then sprayed on the shrimp diet. Different experimental diets were air-dried at room temperature and stored in plastic bags at 4^oC until use.

Experimental animal and set up

The L. vannamei post-larvae (0.5 g) were obtained from a local private shrimp hatchery in Can Tho city, Vietnam. Shrimp were allowed to acclimatize in 2 m³ tank at a salinity of 20 ppt for 7 days before they were subjected for feeding trial with garlic extract. Throughout the acclimatization period, L. vannamei post-larvae were fed a commercial diet three times a day. Thereafter, 1200 shrimp (0.5 g) were randomly allocated into twelve 500 l composite tanks (100 shrimp per tank) in a recirculating aquaculture system. Each experimental group were conducted with three replicates. All tanks were covered with plastic shade net to reduce light intensity and prevent shrimp escape. Shrimp were fed four times daily at 3-5% of body weight per day for 56 days. Uneaten feed was collected 1 h after each meal and dried in an oven at 60^oC. Water temperature, pH, and dissolved oxygen (DO) were measured daily by an HI9828 multi-parameter portable meter (Romania). Alkalinity, hardness, total ammonia nitrogen (TAN), nitrite (NO₂^--N), and nitrate (NO₃^--N) were determined weekly following the standard methods of APHA (2017).

Determination of growth performance and survival rate

At the end of the feeding trial, and survival rate (SR), weigh gain (WG), daily weight gain (DWG), specific growth rate (SGR), biomass (B), and feed conversion ratio (FCR) were calculated according to the following formulae:

Survival rate (%) = (Final numbers/ Initial numbers) × 100

Weight gain (g/shrimp) = Final weight (g) – Initial weight (g)

Daily weight gain (g/day) = (Final weight – Initial weight)/ Days

Specific growth rate (%/day) = ([Ln final wt - Ln initial wt]/ Days) × 100

FCR = Feed intake / Weight gain

B (kg m^-3) = [Final weight × Final population]/Volume of water

Statistical analysis

All data are expressed as mean values ± standard errors (SE). Results were analyzed using one-way analysis of variance (ANOVA), followed by Tukey’s multiple range test to determine differences among treatment. Before analysis, percentages in data were arcsine transformed. The significance level was 0.05. Statistical analysis was performed using SPSS software (version 22).

Results

Water quality parameters

Water quality parameters recorded in the different treatments during the experiment are presented in Table 1. Statistical analysis revealed that no significant differences in temperature, pH, DO, alkalinity and hardness were found among groups (p>0.05). However, TAN in the control group (0 % garlic extract) was significantly higher than the garlic-supplemented treatments at most sampling days (p<0.05). Similarly, the concentrations of NO₂^--N and NO₃^--N were significantly higher for the control group compared to the other groups.

Growth performance and survival of L. vannamei

Shrimp fed with garlic-supplemented diets showed a significantly increased growth rate compared with the control (Table 2). At the end of the experiment, the WG, DWG and SGR of shrimps fed with 1% and 1.5 % garlic were significantly higher than those of shrimps fed with the control and 0.5% (p<0.05) (Figure 1). However, no significant differences in WG, DWG and SGR were observed between the control and 0.5% or between 1% and 1.5% (p>0.05). Similarly, the survival rate and biomass of shrimp were significantly increased by the supplementation of garlic, shrimp fed with 1.5% garlic showed the highest survival rate and biomass. FCR was significantly lower in shrimp fed 1% and 1.5% than in the control (p<0.05) (Table 2, Figure 2).

Figure 1. Effect of garlic on weight gain	Figure 2. Effect of garlic on FCR

Discussion

The utilization of garlic in aquaculture as a therapeutic agent has been developed in recent years. In fact, supplementation of diets with garlic has provided various beneficial effects, including growth promotion, appetite stimulation, immunestimulation, antistress as well as antibacterial in finfish and shrimp larviculture (Büyükdeveci et al 2018; Vaseeharan et al 2011; Millet et al 2011; Aly et al 2010; Nya and Austin 2009; Xie et al 2009; Diab et al 2008; Peyghan et al 2008; Sahu et al 2007; Shalaby et al 2006; Sasmal et al 2005). The results of the present study showed that the growth parameters of L. vannamei were significantly increased when fed supplemented diets containing garlic extract. These results are consistent with the findings from a previous study conducted in giant freshwater prawns, Macrobrachium rosenbergii, which also revealed weight gain and SGR were significantly higher among garlic-supplemented groups compared to the control group (Jahanbakhshi et al 2022). Moreover, the FCR values of shrimp fed garlic extracts (1% and 1.5%) were notably lower than those of the control group, suggesting that the garlic diets may act as an appetizer. This observation is similar to the report of Labrador et al (2016), wherein FCR of the white leg shrimp was fed with diets containing 2%, 4%, and 6% garlic powder. Supa-aksorn et al (2017) reported that garlic containing allicin could stimulate the performance of intestinal flora, thereby enhancing the digestive system and energy utilization, leading to growth improvement.

The mean water quality parameters, including temperature, pH, DO, alkalinity and hardness recorded in different treatments, did not vary significantly and were favorable for the growth of L. vannamei during the culture period. According to Briggs et al (2004), the optimal range of temperature for L. vannamei was from 25-30^oC. The optimal levels of dissolved oxygen and pH for shrimp culture should be 4.5-6 mg/L and 7.5-8.5 mg/L, respectively (Labrador et al 2016). David et al (2004) mentioned that the optimal total alkalinity forL. vannamei should be higher than 75 mg/L. Although TAN, NO₂^- and NO₃^- concentrations in the control group were significantly higher than those in the other groups, these values were within the optimum ranges for shrimp growth. In terms of total ammonia and nitrite, concentrations should not be more than 2 mg/L and 3.5 mg/L, respectively (Chanratchakool 2003; Ferreira et al 2017).

Dietary supplementation with garlic extracts increased the survival rate of L. vannamei compared with the control group. The result obtained appears to agree with the previous study by (Labrador et al 2016), where an improved survival rate of L. vannamei fry was observed upon garlic powder supplementation. A possible explanation could be due to the presence of compounds in garlic such as allin, diallysulphides and allicin (Amagase and Milner 1993). Moreover, Aly et al (2008) indicated that the increased survival rate of garlic powder-supplemented treatments may be a result of the stimulatory effect of garlic on the immune system, which enhances the immune response of aquatic animals.

Conclusion

In conclusion, our results demonstrated dietary garlic extract supplementation could increase the growth performance and survival of L. vannamei. Notably, dietary supplementation with 1.5% garlic extract is estimated to be suitable to support growth and feed utilization in L. vannamei juvenile culture, which provides valuable information for the application of garlic in aquaculture. Further study is needed to clarify the effect of garlic extracts on digestive enzymes as well as the intestinal bacterial community in L. vannamei.

Acknowledgements

The authors acknowledge the financial support of Can Tho University (Code: T2022-122).

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