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Volume 33, Issue 4 (10-2024) |
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Individual and combined effects of the dietary Lactobacillus casei and sodium propionate supplementation on the growth performance and immunity of zebra fish (Danio rerio)
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Ehsan AKBARNEJAD   , Sakineh Yeganeh1 , Sedigheh Mohammadzadeh |
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Abstract: (513 Views) |
Introduction
The emergence of diseases poses significant challenges for the aquaculture sector. While diseases are natural components of aquatic ecosystems, they often lead to economic repercussions for both aquaculture and fisheries. The restrictions on antibiotic use in aquaculture have prompted the development of environmentally friendly alternatives, such as probiotics and organic acids, which serve as immune stimulants. Short-chain fatty acids and organic acids, along with their corresponding salts are emerging as potential alternatives to antibiotic growth promoters in aquaculture, garnering increasing interest. Probiotics, on the other hand, are live microbial feed supplements that help maintain a healthy microbial balance in the host's gut.
Methodology
This study was carried out to evaluate individual and combined effects of the dietary Lactobacillus casei and sodium propionate supplementation on the growth performance and immunity of zebrafish (Danio rerio). A total of 120 zebra fish (335.22 ± 0.23 mg) were placed in 12 tanks with 10 fish per tank and fed with diets containing 0 % Lactobacillus casei and sodium propionate (control, T1) 0.1% Lactobacillus casei (5 x 106 CFU, T2), 1% sodium propionate (T3), and 0.1% Lactobacillus casei bacteria and 1% sodium propionate (T4) for 8 weeks. At the end of the experiment, feeding was halted for 24 hours. Eight fish from each tank were anesthetized with clove oil at a dose of 50 μl/l. After removing the head and tail, the fish were frozen in liquid nitrogen and homogenized using a manual homogenizer. The resulting homogenate was then prepared in 25 mM Tris-HCl buffer (pH 7.2) for the measurement of immune factors.
Results
The experimental diets significantly affected the final weight and weight gain of the fish. Fish fed with Lactobacillus casei and sodium propionate had a higher weight compared to the control group (Table 1, p<0.05). The specific growth rate also varied among the diets, with the lowest growth rate observed in the control group and the highest in the groups receiving sodium propionate and Lactobacillus casei, which differed significantly from the control group (p<0.05). No significant differences were found among the treatments in triglyceride, cholesterol, and glucose levels (Table 2, p<0.05). Serum albumin analysis indicated no significant difference between treatments 1 and 3, nor between treatments 2 and 4 (Table 2, p<0.05). However, significant differences were noted between treatments 1 and 3 and treatments 2 and 4, with the highest levels found in treatments 2 and 4. Regarding the globulin index, there was no significant difference between the control group and treatment 3, but significant differences were observed with the other two treatments (Table 2, p<0.05). The highest activities of superoxide dismutase, catalase, and glutathione peroxidase were recorded in treatment 4, which significantly differed from the control group (p<0.05) but showed no significant difference from treatment 3 (Table 3, p<0.05). Significant differences were also observed in total serum protein and immunoglobulin levels, with the highest levels in treatments containing Lactobacillus casei and sodium propionate (treatments 2, 3, and 4), which differed significantly from the control group (Table 4, p<0.05). Notably, significant variations were found in lysozyme and complement levels among the treatments, with the highest levels in treatments 3 and 4 (Table 4, p<0.05).
Table 1: Growth performance in zebrafish fed with experimental diets for 8 weeks (mean ± SD)
| Fourth treatment |
Third treatment |
Second treatment |
First treatment |
Factors/treatments |
| 458.3 ± 13.1a |
450.1 ± 11.2b |
432.9 ± 13.4b |
387.6 ± 9.4c |
Final weight (mg) |
| 123.9 ± 14.9a |
114.6 ± 10.2b |
96.9 ± 15.7b |
52.4 ± 10.1c |
Weight gain (mg) |
| 37.1 ± 4.7a |
34.2 ± 2.5ab |
28.8 ± 4.8b |
15.6 ± 3.0c |
Body weight gain (%) |
| 0.22 ± 0.02a |
0.21 ± 0.01a |
0.18 ± 0.02b |
0.1 ± 0.01c |
Specific growth rate (%/day) |
| 100 |
100 |
100 |
100 |
Survival rate (%) |
Different letters indicate significant differences between groups (mean ± SD).
Table 2: Biochemistry index in in zebrafish fed with experimental diets for 8 weeks (mean ± SD)
| Fourth treatment |
Third treatment |
Second treatment |
First treatment |
Factors/treatments |
| 79.9 ± 1.6a |
88.1 ± 5.4a |
83.85 ± 3.65a |
78.6 ± 1.1a |
Triglyceride (mg/100g tissue) |
| 174.5 ± 4.5a |
166.5 ± 11.5a |
181.5 ± 8.8a |
156.8 ± 11.2a |
Glucose (mg/100 g tissue) |
| 24.85 ± 2.35a |
25.6 ± 0.7a |
24.8 ± 0.4a |
23.8 ± 0.3a |
Cholesterol (mg/ 100 g tissue) |
| 17.05 ± 0.25a |
14.6 ± 0.1b |
17.20 ± 0.3a |
13.59 ± 0.65b |
Albumin (mg/ 100 g tissue) |
| 60.9 ± 0.2b |
58.55 ± 0.25c |
64.35 ± 0.65a |
58 ± 0.4c |
Globulin (mg/ 100 g tissue) |
Different letters indicate significant differences between groups (mean ± SD).
Table 3. Antioxidant enzymes in in zebrafish fed with experimental diets for 8 weeks (mean ± SD)
| Fourth treatment |
Third treatment |
Second treatment |
First treatment |
Factors/treatments |
| 69 ± 7.1a |
67.0 ± 0.1a |
65.6 ± 0.1a |
53.5 ± 0.6b |
SOD (u/g) |
| 72.8 ± 0.3a |
73.2 ± 3.5a |
63.85 ± 0.4b |
63.3 ± 0.4b |
CAT (u/g) |
| 235 ± 0.3a |
232 ± 3a |
223.5 ± 1.5b |
221.1 ± 2.5b |
GPx (u/g) |
Different letters indicate significant differences between groups (mean ± SD).
Table 4: immunity index in zebrafish in zebrafish fed with experimental diets for 8 weeks (mean ± SD)
| Fourth treatment |
Third treatment |
Second treatment |
First treatment |
Factors/treatments |
| 87.95 ± 0.45a |
83.15 ± 0.35a |
82.55 ± 0.95a |
71.95 ± 0.25b |
Total protein (mg/100g tissue) |
| 1.46 ± 0.15a |
1.44 ± 0.15a |
1.48 ± 0.01a |
1.4 ± 0.01b |
Immunoglobulin (mg/100 g tissue) |
| 40.95 ± 0.25a |
38.05 ± 1.45a |
20.5 ± 3.2b |
22.70 ± 1.1c |
Lysozyme (u/g/min) |
| 105.1 ± 1.5a |
89.5 ± 0.5b |
84.5 ± 1.5b |
86.5 ± 0.5b |
ACH50 (U%) |
Different letters indicate significant differences between groups (mean ± SD).
Discussion and conclusions
Fish growth and immunity improved in fish fed supplemented diets compare to the control group. Improving fish performance through organic acids and probiotics is possible in several ways, including reducing gastric pH, which increases pepsin activation, reducing intestinal pH, which leads to increased mineral dissolution and subsequent absorption by organic and probiotics. It can be concluded that the use of a diet containing Lactobacillus casei and sodium propionate in zebrafish has numerous positive effects. Overall, the findings suggest that these supplements can be effectively utilized both separately and in combination, specifically at levels of 1% sodium propionate and 0.1% Lactobacillus casei (5 × 106 CFU) in aquaculture.
Conflicts of interest
The authors declare no conflicts of interest. |
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| Keywords: Lactobacillus casei, sodium propionate, zebrafish, immunity, enzyme, biochemical index |
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Full-Text [PDF 706 kb]
(112 Downloads)
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Type of Study: Research |
Subject:
تغذيه
Received: 2024/07/4 | Accepted: 2024/10/31 | Published: 2024/12/23
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aKBARNEJAD E, Yeganeh S, mohammadzadeh S. Individual and combined effects of the dietary Lactobacillus casei and sodium propionate supplementation on the growth performance and immunity of zebra fish (Danio rerio). isfj 2024; 33 (4) :131-142 URL: http://isfj.ir/article-1-2817-en.html
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