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Article

Effects of Bacillus licheniformis Feeding on the Growth Performance, Blood Parameters and Intestinal Microbiota of Adult Hybrid Sturgeon

by
Quansen Xie
1,2,*,
Yu Wang
3,
Xinyu Huang
3,
Yiran Liu
1,
Mingjian Yang
1,
Haochun Xing
1,
Caimei Yang
2,
Caihong Hu
4,
Mingzhu Pan
3 and
Zhitao Qi
3,*
1
School of Life Science and Engineering, Handan University, Handan 056005, China
2
Postdoctoral Workstation of Zhejiang Huijia Biotechnology Co., Ltd., Huzhou313000, China
3
School of Marine and Bioengineering, Yancheng Institute of Technology, Yancheng 224000, China
4
School of Animal Science and Technology, Zhejiang University, Hangzhou 310027, China
*
Authors to whom correspondence should be addressed.
Fishes 2025, 10(5), 189; https://doi.org/10.3390/fishes10050189
Submission received: 21 March 2025 / Revised: 16 April 2025 / Accepted: 18 April 2025 / Published: 22 April 2025
(This article belongs to the Special Issue Dietary Supplementation in Aquaculture)
Browse Figure
Versions Notes

Abstract

:
In this study, we added Bacillus licheniformis to the diet of hybrid sturgeon (Acipenser baerii ♀ Acipenser schrenkii ♂) to determine its effects on growth performance, blood physical and chemical indices and intestinal microflora composition. One hundred and sixty adult hybrid sturgeon were selected and fed with four types of diets (equal nitrogen and fat levels) that were respectively supplemented with 0.00% (control group), 0.10% (Group B), 0.20% (Group C) and 0.40% (Group D) B. licheniformis for 120 days. Results showed that the fish in group C had the highest final body weight, weight gain rate and specific growth rate (p < 0.05). The feed coefficients, crude protein and crude fat of group B, group C and group D were significantly lower than that of group A (p < 0.05). And the crude protein (CP) and crude fat (EE) in groups B, C and D were significantly higher than the control group (p < 0.05). The serum TC and TG, ALP, ALT, AST and GLU contents in the B. licheniformis-added groups were also significantly higher than those of the control group (p < 0.05). In addition, Cetobacterium was the dominant bacterial taxon in each group. With increasing the content of B. licheniformis in the diet, the Cetobacterium content decreased and the Plesiomonas content increased correspondingly. Adding B. licheniformis to the diet greatly decreased the abundance of Streptococcus, Candidatus Competibacter and Lactococcus. Our results indicated that appropriately adding (0.20%) B. licheniformis could increase growth, reduce the feed coefficient and increase the diversity of the intestinal microbiota of hybrid sturgeon.
Keywords:
Bacillus licheniformis; sturgeon; growth performance; intestinal microbiota
Key Contribution: Appropriately adding (0.20%) B. licheniformis to the diet can increase the growth, reduce the feed coefficient and increase the diversity of the intestinal microbiota of hybrid sturgeon.

1. Introduction

Sturgeon, the oldest and most primitive cold-water fish on Earth, is known as a “living fossil” and has important economic and scientific research values [1]. It is widely cultured for its delicious meat and rich nutritional content. China is the largest sturgeon producer, accounting for 86% of the total sturgeon production worldwide [2,3], and mainly breeds hybrid sturgeons. Among them, A. schrenckii has a fast growth rate, but poor disease resistance, intolerance to transportation and difficulty in domestication. The Acipenser baerii, on the other hand, grows slowly but has strong disease resistance and transportation tolerance. After crossbreeding Siberian sturgeon and Chinese sturgeon, their hybrid offspring showed superior adaptability, growth rates, disease resistance and survival rates during transportation compared to their parents [4]. However, more and more reports on diseases occurred with the expansion of intensive culture models, which has become a bottleneck problem restricting the healthy development of the sturgeon culture industry [5,6,7].
Probiotics are beneficial microorganisms that can colonize the intestines of aquatic animals or improve aquaculture water quality. They can enhance the growth performance of aquatic animals and maintain the aquaculture environment, and they usually do not have a negative impact on consumers [8]. Multiple studies have shown that Lactococcus, Saccharomyces, Lactobacillus and Enterococcus are commonly used in aquaculture to alleviate the pressure of intensive aquaculture and are considered effective methods for promoting sustainable development of aquaculture [9,10,11]. Probiotic preparations can be metabolized and digested in the digestive tract of fish to produce organic acids, amino acids and other substances, which can promote fish growth and fish immunity [12]. The most direct and efficient way to administer probiotic preparations to fish is to add them directly to feed as additives.
With the increase in the global population and the continuous expansion of demand for aquatic products, the use of antibiotics in fish is becoming increasingly widespread. However, excessive use of antibiotics has also brought a series of problems, such as antibiotic residues, increased drug resistance, water pollution, etc. [9]. Probiotics play a crucial role in enhancing the overall sustainability of sturgeon farming and aquaculture, such as reducing stress, promoting digestion, improving growth and water quality, increasing the survival rate of aquatic animals after infection, reducing parasite infestation, and minimizing the environmental impact of aquaculture [13,14]. Bacillus subtilis is an effective alternative to antibiotics [15]. Among Bacillus species, B. licheniformis is the most promising, which can easily form spores and be added to aquatic feed to degrade large particulate nutrients in the feed [16,17,18]. It has been found that Bacillus licheniformis could promote disease resistance and growth in fish. For example, dietary fructooligosaccharide and B. licheniformis increased the innate immunity, antioxidant capability and disease resistance against Aeromonas hydrophila of triangular bream (Megalobrama terminalis) [19]. Also, the administration of B. licheniformis could enhance the resistance of rainbow trout (Oncorhynchus mykiss) against the Yersinia ruckeri challenge [20]. At present, there is not much research on the application of Bacillus licheniformis in sturgeon. In the present study, B. licheniformis was added to the diets of sturgeons, and its effects on growth performance, blood physicochemical indicators and gut microbiota were evaluated. Blood is an important tissue in the animal body that is closely related to the body’s metabolism, nutritional status and diseases. A review of the research on blood indicators in fish was conducted by analyzing physiological and biochemical indicators of fish blood. Our results provide a basis for the administration of B. licheniformis in the feed of sturgeon.

2. Materials and Methods

2.1. Bacillus licheniformis and Diet Preparation

Table 1 shows the composition and nutritional level of the basic diet. The B. licheniformis, provided by Zhejiang Huijia Biotechnology Co., Ltd. (Hangzhou, China), was incorporated into the basic diet at ratios of 0.00% (0 g/kg basic diet, control group or group A), 0.10% (1 g/kg basic diet, group B), 0.20% (2 g/kg basic diet, group C) and 0.40% (4 g/kg basic diet, group D) (Table 1). During the feed preparation, each raw material was finely crushed and then accurately weighed and mixed according to the designed formula to prepare feed pellets with a diameter of 1.5 mm. The mixture was placed in an oven (60 °C) for 45 min. After air drying in a cool indoor location, the feed pellets were divided into sealed bags and stored at −20 °C for use. The temperature during the entire preparation process was less than 100 °C.

2.2. Fish and Fish Culture

Two hundred adult hybrid sturgeon (Acipenser baerii, Siberian sturgeon, ♀ A. schrenckii ♂) were purchased from a sturgeon culture farm in Quzhou City, Zhejiang Province, China. The fish were transported to the sturgeon breeding base in Kecheng District, Quzhou City, for temporary cultivation for 7 days. Hybrid sturgeon were measured with a conventional electronic scale (5 kg/0.01 g) and soft ruler (2 m/0.1 cm). The fish with an initial body weight of 850.15 ± 15.00 g were selected and randomly divided into 4 groups, with each group containing 4 replicates (10 hybrid sturgeon each replicate). Each group of sturgeons was raised in a circular bucket (a diameter of 2 m and a volume of 2.51 m3, exchange rate of 12 L·min−1, quantity of water is 2.26 L). After feeding the fish with a basic diet for 1 week, the formal experiment started under natural light/dark, water temperature of (15 ± 2) °C, dissolved oxygen > 6 mg/L, and ammonia nitrogen < 0.1 mg/L. Feed was placed at designated positions twice a day (8:00 and 15:00) with a feeding rate of 3% to 5% fish body weight. The remaining feed was collected in time after feeding. The whole experiment lasted for 120 days.

2.3. Measurement of Growth Indicators

The formulas for calculating growth performance were as follows:
Weight gain (WG, %) = 100 × (Wt − W0)/W0
Specific growth rate (SGR, %·d−1) = 100 × (lnWt − lnW0)/t
Feed conversion ratio (FCR) = F/(Wt − W0)
where W0 and Wt are the initial and final body masses (g) of the experimental hybrid sturgeon, respectively; F is the feed intake (g) and t is the experimental time (d).
Condition factor (CF, g·cm−3) = 100 × final body mass/body length3
Viscerosomatic index (VSI, %) = 100 × visceral weight/terminal body mass
Hepatosomatic index (HSl, %) = 100 × liver weight/terminal body mass

2.4. Measurement of Blood Biochemical Indicators

At the end of the feeding experiment, 10 fish were randomly selected from each group, and 10 mL of tail artery blood was collected via a sterile syringe and placed in a 10 mL centrifuge tube. The mixture was incubated at 4 °C for 12 h, the serum was allowed to precipitate, the mixture was centrifuged for 15 min at −1 °C and the serum was collected; the blood biochemical indicators, including albumin (ALB), alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), serum glucose (GLU), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), total serum cholesterol (TC), triglyceride (TG) and total protein (TP), were measured by an automated biochemical analyzer (Hitachi LABOSPECT 008 AS, Shenzhen New Industry Biomedical Engineering Co., Ltd., Shenzhen, China). The use of animals in this study was approved by the College of Life Sciences and Engineering, Handan University (HDXY SGXY2024-03), which was carried out according to the guidelines for the care and use of experimental animals.

2.5. Diversity of the Gut Microbiota

At the end of the experiment, fecal samples were randomly collected from eight hybrid sturgeon in each group, processed using the QIAGEN reagent kit, and subsequently stored in cryotubes. The fecal samples were stored in liquid nitrogen for genomic DNA amplification and high-variability region sequencing (primers 319F and 806R) on the Illumina MiSeq 2 × 300 bp paired-end sequencing platform at Hangzhou Lianchuan Biotechnology Co., Ltd. The uniformity of the distributions of different bacterial communities at the phylum level was statistically analyzed.

2.6. Statistical Analysis

The experimental data are expressed as the mean ± standard error (mean ± SD). All data were subjected to one-way ANOVA by Statistica 16.0 software, and Duncan’s method was used for intergroup multiple comparisons, with a significance level of 0.05.

3. Results

3.1. Effects of B. licheniformis on the Growth Performance and Body Indices of Sturgeon

As shown in Table 2, the survival rate of sturgeon in each group was over 90%. The final body weight, weight gain rate and specific growth rate tended to first increase and then decrease with the increasing addition of B. licheniformis. The fish in group C had the highest final body weight, weight gain rate and specific growth rate (p < 0.05). The feed coefficients of group B, group C and group D were significantly lower than that of the control group (group A) (p < 0.05). The feed coefficient in group C reached 1.46, which was the lowest compared with that of groups B and D. There was no significant difference in the organ/body ratio and liver/body ratio among groups B, C and D (p > 0.05).

3.2. Effects of B. licheniformis on the Body Composition of Hybrid Sturgeon

As demonstrated in Table 3, hybrid sturgeon in groups B, C and D exhibited significantly higher levels of crude protein (CP) and ether extract (EE) compared to the control group (p < 0.05). No differences were observed for CP and EE in groups B, C and D. There was no significant difference in ash and water content among each group (p > 0.05).

3.3. Effects of B. licheniformis on the Blood Biochemical Parameters of Sturgeon

It was found that adding B. licheniformis to sturgeon diets could significantly increase the serum TC and TG contents when compared with that of the control group (Table 4) (p < 0.05). In addition, serum ALP, ALT, AST and GLU contents in the B. licheniformis-added groups were also significantly higher than that of the control group (p < 0.05). There was no significant difference in total protein among each group (p > 0.05).

3.4. Effects of B. licheniformis on the Intestinal Microbiota of Sturgeon

Table 5 shows the diversity indices of the intestinal microbiota samples in each group. The OUT and ACE indexes in group B, C and D were significantly lower than that of the control group (p < 0.05). The Chao1, Simpson and Shannon indices in groups B, C and D were significantly higher than that of the control group (p < 0.05). Group C had the highest Chao1, Simpson and Shannon indices, which respectively reached 279.30, 0.87 and 0.79 (Table 5).
Table 6 shows the distribution of intestinal microbial species at the genus level. Cetobacterium was the dominant bacterial taxon in each group. With increasing the contents of B. licheniformis in the diet, the Cetobacterium content decreased and the Plesiomonas content increased correspondingly. Group D had the lowest of the Cetobacterium content and the highest of the Plesiomonas content. Adding B. licheniformis to the diet greatly decreased the abundance of Streptococcus, Candidatus Competibacter and Lactococcus (Table 6). The abundance of Clostridium, Lactobacillus and Bacillus increased with the addition of B. licheniformis, and the highest abundance was observed in group D (Table 6, Figure 1).

4. Discussion

Composite probiotics are considered healthy, green and safe products with the potential to replace antibiotics [25]. Probiotic supplementation in feed could effectively promote the growth and survival of animals [26]. In recent years, Bacillus species have become commonly used probiotics as feed additives. Research on aquatic animals has shown that the addition of Bacillus to the diet promoted the growth of sturgeon [17,18,19]. Gao et al. added different doses of Bacillus powder to the formula feed for juvenile Siberian sturgeon, revealing that the group supplemented with 0.10% Bacillus powder presented the maximum weight gain rate and specific growth rate [25]. The addition of Bacillus powder to the feed promoted the growth of Siberian sturgeon. Zhang et al. evaluated the effect of feeding B. subtilis to hybrid sturgeon juveniles, finding that there was no significant difference in specific growth rates between the B. subtilis-added group and the control group [27]. The main modes of action of Bacillus licheniformis and Bacillus subtilis for antibacterial and disease prevention are different. During the process of growth metabolism, Bacillus subtilis can produce active substances such as Bacillus subtilis, polymyxin, nystatin and clarithromycin, all of which can clearly limit the growth of pathogens or endogenous pathogenic bacteria [28]. After entering the animal intestine in the form of endospores, Bacillus licheniformis can quickly revive and secrete various enzymes such as protease, lipase, amylase, etc. in the upper part of the intestine [29,30]. These enzymes break down feed components that are difficult for the host to digest into small molecule substances that can be absorbed and utilized by the host, promoting the digestion and absorption of nutrients [31]. In this study, we found that the final body weight, weight gain rate and specific growth rate of the B. licheniformis-added groups tended to first increase but then decrease with increasing the B. licheniformis, which reached the highest in group C (0.20% B. licheniformis), indicating that B. licheniformis could promote the growth of adult hybrid sturgeon. Also, the growth of hybrid sturgeon did not increase with the increase in B. licheniformis content. Similar results were also observed in Epinephelus fuscoguttatus ♀ E. lanceolatus ♂ hybrid and Rachycentron canadum [32], indicating that adding an appropriate proportion of probiotics in the diet could better promote the growth of fish species.
The compositions of the diet significantly affect the composition and proportion of nutritional components of fish body and muscle, such as crude protein, fat and ash [33]. The effects of probiotics on the nutritional composition of fish muscles and body varied. Zhou et al. reported that adding lactic acid bacteria to the diet of tilapia could increase the crude protein content of fish bodies and reduce the water content [34]. Hu et al. revealed that adding probiotics to the diet of eel had no effect on crude protein and fat levels but significantly increased the content of crude ash [35]. The addition of probiotics to the diet of turbot significantly increased the crude protein content in the fish body and muscle, whereas it had no effect on the water, crude fat and crude ash [36]. We found that with the increasing amount of B. licheniformis in the diet, the MS, CP, EE and ash contents of the hybrid sturgeon bodies tended to increase first and then decrease. The CP and EE levels in the B. licheniformis-added groups were significantly higher than those in the control group. The effects of probiotics on the nutritional composition of fish bodies and muscles might depend on the type of animals, feeding environment and the vitality of probiotics [37]. Moreover, the addition of probiotics to the diet increased the protein content in the fish body and muscle, which may be due to the promotion of protein absorption by probiotics [38].
Changes in physical and chemical indicators in blood can reflect physiological changes in the body of animals [39]. Blood plays a crucial role in maintaining the homeostasis of the internal environment in animals, and serum biochemical indicators, to a certain extent, reflect the body’s ability to metabolize various nutrients and nonspecific immunity [40]. The TP content in serum can, to some extent, reflect the ability of animals to metabolize proteins, whereas the TC and TG contents can reflect the lipid content in animal serum [32]. Shen et al. reported that adding B. subtilis to the diet could increase the activity of serum superoxide enzymes and the total antioxidant capacity [41]. And, adding B. subtilis to the feed of Japanese flounder could significantly increase the plasma GLU content [42]. Our study revealed that adding 0.10–0.40% B. licheniformis to hybrid sturgeon feed could significantly increase the serum GLU content, indicating that this Bacillus species could enhance the ability of the sturgeon to utilize and transform GLU, which was consistent with the results of Pan et al. [35]. Also, B. licheniformis could improve the antioxidant capacity and lysozyme activity of hybrid adult sturgeon and improve animal immune function by stimulating cellular and humoral immunity [43]. However, Bacillus species have specificity for different animals, and the characteristics of its strain and safety for fish remain to be further verified.
The morphology and tissue structure of the intestine are extremely important for growth, development, nutrient digestion and nutrient absorption in aquatic animals [44,45]. The intestinal villi promote the full utilization and absorption of nutrients by increasing the surface area of the intestine [46]. The intestinal microbiota depends on or restricts the health of the host, and changes in the structure of the intestinal microbiota can regulate growth, immunity and other aspects of animal physiology [47]. Therefore, establishing a normal intestinal microbiota is extremely important [34,48]. It has been found that probiotics, as feed additives, have varying impacts on growth and development, disease resistance, maintenance of the intestinal ecological balance and feed utilization efficiency in fish [3,49,50,51]. Previous studies have shown that adding B. subtilis to feed could improve the intestinal microbiota composition of grass carp (Ctenopharyngodon idella) [52]. Our study revealed that the sturgeon in the group with 0.20% B. licheniformis addition presented an improvement in the intestinal microbiota, exhibiting an increase in intestinal microbial diversity and an increase in the Bacillus content in the intestine. As the B. licheniformis proportion increased to 0.40%, the diversity of the intestinal microbiota significantly decreased, and the proportion of Pseudomonas bacteria significantly increased, which could cause diarrhea in fish.

5. Conclusions

In summary, feeding hybrid adult sturgeon with different doses of B. licheniformis could significantly reduce the feed coefficient and increase the ability of sturgeon to utilize and transform GLU and fat. When the amount of B. licheniformis added was 0.20%, the microecological environment of the hybrid sturgeon intestine was improved, and the digestive and absorptive functions of the intestine also increased. This addition significantly increased the diversity of the intestinal microbiota in hybrid sturgeon.

Author Contributions

Methodology, Q.X. and Y.L.; data curation, M.Y., X.H. and H.X.; resources, C.Y. and C.H.; writing—original draft preparation, Q.X. and M.P.; writing—review and editing, Z.Q.; validation, Y.W. All authors have read and agreed to the published version of the manuscript.

Funding

The study was supported by the Funding Project for Handan University: Innovative Team for Freshwater Aquaculture in the Southern Hebei Region (Grant No. XKYTD202301).

Institutional Review Board Statement

The use of animal in this study was approved by college of Life Sciences and Engineer, Handan University (HDXY SGXY2024-03),which was carried out according to the guidelines for the carried out according to the guidelines for the care and use of experimental animals.

Informed Consent Statement

Not applicable.

Data Availability Statement

Data will be made available on request.

Conflicts of Interest

Authors Xie Quansen and Yang Caimei were employed by the company Zhejiang Huijia Biotechnology Co., Ltd., Huzhou, China. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

References

  1. Pikitch, E.K.; Doukakis, P.; Lauck, L.; Chakrabarty, P.; Erickson, D.L. Status, trends and management of sturgeon and paddlefish fisheries. Fish Fish. 2005, 6, 233–265. [Google Scholar] [CrossRef]
  2. Chen, X.H.; Li, C.J.; Yang, C.G.; Zhang, S.H.; Wu, J.P. Status and prospects of techniques in the sturgeon aquaculture industry in China. Freshw. Fish. 2017, 47, 108–112. [Google Scholar]
  3. Zhang, Y.; Sun, D.J.; Wang, B.; Xia, Y.T.; Wu, W.; Xu, S.J.; Liu, X.Y. Hybrid sturgeon “Xunlong no. 1”. China Fish. 2017, 5, 62–67. (In Chinese) [Google Scholar]
  4. Chen, X.; Wei, Z. Research progress on hybrid sturgeon in China. Sichuan Agric. Sci. Technol. 2023, 10, 122–124. [Google Scholar]
  5. Padeniya, U.; Larson, E.T.; Septriani, S.; Pataueg, A.; Kafui, A.R.; Hasan, E.; Brown, C.L. Probiotic treatment enhances pre-feeding larval development and early survival in zebrafish Danio rerio. J. Aquat. Anim. Health 2022, 34, 3–11. [Google Scholar] [CrossRef]
  6. Zhang, S.; Huang, J.; Di, J.; Du, H.; Xu, Q.; Zhou, Q.; Wei, Q. The genome sequence of a new strain of Mycobacterium ulcerans ecovar Liflandii, emerging as a sturgeon pathogen. Aquaculture 2018, 489, 141–147. [Google Scholar] [CrossRef]
  7. Di, J.; Zhang, S.; Huang, J.; Du, H.; Zhou, Y.; Zhou, Q.; Wei, Q. Isolation and identification of pathogens causing haemorrhagic septicaemia in cultured Chinese sturgeon (Acipenser sinensis). Aquac. Res. 2018, 49, 3624–3633. [Google Scholar] [CrossRef]
  8. Dawood, M.A.; Koshio, S.; Ishikawa, M.; Yokoyama, S.; El Basuini, M.F.; Hossain, M.S.; Wei, H. Dietary supplementation of β-glucan improves growth performance, the innate immune response and stress resistance of red sea bream, Pagrus major. Aquac. Nutr. 2017, 23, 148–159. [Google Scholar] [CrossRef]
  9. Ringø, E.; Hoseinifar, S.H.; Ghosh, K.; Doan, H.V.; Beck, B.R.; Song, S.K. Lactic acidbacteria in finfish—An update. Front. Microbiol. 2018, 9, 1818. [Google Scholar] [CrossRef]
  10. Wang, Y.B.; Xu, Z.R. Effect of probiotics for common carp (Cyprinus carpio) based on growth performance and digestive enzyme activities. Anim. Feed. Sci. Technol. 2006, 127, 283–292. [Google Scholar]
  11. Ringø, E. Evaluation of probiotic strain Bacillus subtilis C-3102 as a feed supplement for koi carp (Cyprinus carpio). J. Aquac. Res. Dev. 2011, S1, 5. [Google Scholar]
  12. Samat, N.A.; Yusoff, F.M.; Rasdi, N.W.; Karim, M. The efficacy of Moina micrura enriched with probiotic Bacillus pocheonensis in enhancing survival and disease resistance of red hybrid tilapia (Oreochromis spp.) larvae. Antibiotics 2021, 10, 989. [Google Scholar] [CrossRef] [PubMed]
  13. Dawood, M.A.; Koshio, S.; Abdel-Daim, M.M.; Van Doan, H. Probiotic application for sustainable aquaculture. Rev. Aquac. 2019, 11, 907–924. [Google Scholar] [CrossRef]
  14. Onomu, A.J.; Okuthe, G.E. The role of functional feed additives in enhancing aquaculture sustainability. Fishes 2024, 9, 167. [Google Scholar] [CrossRef]
  15. Jiang, M.; Zhang, W.; Luo, L.; Tang, X.; Li, Y.; Yu, X.; Wu, Z. Effects of Bacillus methylotrophicus WM-1 on water quality dynamics and cultural fish in grass carp culture system. Aquac. Res. 2021, 52, 3371–3380. [Google Scholar] [CrossRef]
  16. Li, X.Y.; Kong, J.; Zhao, F.; Yang, M.J. Application and research progress of probiotics in sturgeon feed additive. Guizhou Anim. Sci. Vet. Med. 2016, 40, 61–63. (In Chinese) [Google Scholar]
  17. Alizadeh, A.M.; Hosseini, H.; Meybodi, N.M.; Hashempour-Baltork, F.; Alizadeh-Sani, M.; Tajdar-Oranj, B.; Pirhadi, M.; Khaneghah, A.M. Mitigation of potentially toxic elements in food products by probiotic bacteria: A comprehensive review. Food Res. Int. 2022, 152, 110324. [Google Scholar] [CrossRef]
  18. Li, B.; Yang, J.; Huang, J.H.; Han, W.L.; He, H.C. The effects of three probiotics on the growth and immune system of hybrid sturgeon (Huso dauricus ♀ × Acipenser schrenckii). Freshw. Fish. 2014, 1, 78–83. [Google Scholar]
  19. Zhang, C.N.; Li, X.F.; Xu, W.N.; Jiang, G.Z.; Lu, K.L.; Wang, L.N.; Liu, W.B. Combined effects of dietary fructooligosaccharide and Bacillus licheniformis on innate immunity, antioxidant capability and disease resistance of triangular bream (Megalobrama terminalis). Fish Shellfish. Immunol. 2013, 35, 1380–1386. [Google Scholar] [CrossRef]
  20. Raida, M.K.; Larsen, J.L.; Nielsen, M.E.; Buchmann, K. Enhanced resistance of rainbow trout, Oncorhynchus mykiss (Walbaum), against Yersinia ruckeri challenge following oral administration of Bacillus subtilis and B. licheniformis (BioPlus 2B). J. Fish Dis. 2003, 26, 495–498. [Google Scholar] [CrossRef]
  21. GB 5009.5-2016; National Food Safety Standard Determination of Protein in Food. National Health and Family Planning Commission of the People’s Republic of China: Beijing, China, 2016.
  22. GB 5009.6-2016; National Food Safety Standard Determination of Fat in Food. National Health and Family Planning Commission of the People’s Republic of China: Beijing, China, 2016.
  23. GB 5009.4-2016; National Food Safety Standard Determination of Ash in Food. National Health and Family Planning Commission of the People’s Republic of China: Beijing, China, 2016.
  24. GB 5009.3-2016; National Food Safety Standard Determination of Moisture in Food. National Health and Family Planning Commission of the People’s Republic of China: Beijing, China, 2016.
  25. Gao, X.; Gai, L.Q.; Li, M.Y.; Li, M.Y.; Guo, X.X.; An, R.Y. Effects of Bacillus spp. on the growth performance and intestine bacteria in juvenile Acipenser baeri. J. Hebei Norm. Univ. 2009, 33, 377–382. [Google Scholar]
  26. Olmos, J.; Acosta, M.; Mendoza, G.; Pitones, V. Bacillus subtilis, an ideal probiotic bacterium to shrimp and fish aquaculture that increase feed digestibility, prevent microbial diseases, and avoid water pollution. Arch. Microbiol. 2020, 202, 427–435. [Google Scholar] [CrossRef] [PubMed]
  27. Zhang, S.H.; Wu, J.P.; Chu, Z.P.; Xu, Q.Q.; He, J.Y.; Jin, J.L.; Chen, X. Effects of feeding Bacillus subtilis on growth and intestinal microflora of hybrid sturgeon. Mar. Fish. 2021, 43, 71–80. [Google Scholar] [CrossRef]
  28. Ji, M.; Yao, K.; Li, G.; Wu, X.; Chen, H.; Zhuang, Y. Control effects of Bacillus subtilis DJ-6 and pyraclostrobin alone and in combination against Fusarium oxysporum. Agric. Sci. Technol. 2014, 15, 2020–2025. [Google Scholar] [CrossRef]
  29. Cao, Y.; Li, Z.; Feng, J.; Chen, Y.Q.; Wu, Z.H. Study on the digestive activity of extracellular products of Bacillus licheniformis. J. Trop. Oceanogr. 2005, 6, 6–12. [Google Scholar]
  30. Sangeetha, R.; Geetha, A.; Arulpandi, I. Concomitant production of protease and lipase by Bacillus licheniformis VSG1: Production, purificationand characterization. Braz. J. Microbiol. 2010, 41, 179–185. [Google Scholar] [CrossRef]
  31. Xu, Y. Research Progress on Application of Bacillus licheniformis in aquaculture. Fish. Res. 2018, 40, 83–88. [Google Scholar] [CrossRef]
  32. Sanders, M.E.; Merenstein, D.J.; Reid, G.; Gibson, G.R.; Rastall, R.A. Probiotics and prebiotics in intestinal health and disease: From biology to the clinic. Nat. Rev. Gastroenterol. Hepatol. 2019, 16, 605–616. [Google Scholar]
  33. Ahmed, I.; Jan, K.; Fatma, S.; Dawood, M.A.O. Muscle proximate composition of various food fish species and their nutritional significance: A review. J. Anim. Physiol. Anim. Nutr. 2022, 106, 690–719. [Google Scholar] [CrossRef]
  34. Yi, Z.; Liu, X.; Liang, L.; Wang, G.; Xiong, Z.; Zhang, H.; Song, X.; Ai, L.; Xia, Y. Antrodin A from Antrodia camphorate modulates the gut microbiome and liver metabolome in mice exposed to acute alcohol intake. Food Funct. 2021, 12, 2925–2937. [Google Scholar] [CrossRef]
  35. Cheng, Y.; Huang, K.; Huang, X.Y.; Zhong, L.; Wu, L.; Huang, Q.; Tang, L. Effects of dietary Bacillus subtilis on growth performance, immunity and antioxidation function of juvenile genetic improvement of farmed tilapia (GIFT, Oreochromis niloticus). Chin. J. Anim. Nutr. 2014, 1503–1512. [Google Scholar]
  36. Liu, Y.; Zhao, W.X.; Yue, X.L.; Sun, X.; Guan, S.G.; Yu, C.Y. Effects of compound feeds on growth performance of Scophthalmus Maximus at different growth stages. J. Aquac. 2014, 45, 335–492. [Google Scholar]
  37. Wang, A.; Ran, C.; Wang, Y.; Zhang, Z.; Ding, Q.; Yang, Y.; Olsen, R.E.; Ringø, E.; Bindelle, J.; Zhou, Z. Use of probiotics in aquaculture of China—A review of the past decade. Fish Shellfish. Immunol. 2019, 86, 734–755. [Google Scholar] [CrossRef] [PubMed]
  38. Ding, H.; Huang, P.; Zhang, W.M.; He, J.H.; Kong, X.F. Effects of dietary Bacillus subtilis on growth performance and plasma biochemical parameters of nursery piglets. Chin. J. Anim. Nutr. 2020, 32, 1–8. [Google Scholar]
  39. Qiao, Z.G.; Jia, W.; Zhang, X.G.; Shen, F.F. Differences in growth, blood physiology and biochemical components between female and male catfish Silurus asotus. J. Dalian Ocean. Univ. 2019, 34, 376–380. [Google Scholar]
  40. Zhu, P.; Tang, Y.; Fan, J.; Fang, J.; Peng, X.; Cui, H. Hematological parameters and blood cell morphology of male and female Schizothorax (Racoma) davidi (Sauvage). J. World Aquac. Soc. 2017, 48, 821–830. [Google Scholar] [CrossRef]
  41. Shen, W.-Y.; Fu, L.-L.; Li, W.-F.; Zhu, Y.-R. Effect of dietary supplementation with Bacillus subtilis on the growth, performance, immune response and antioxidant activities of the shrimp (Litopenaeus vannamei). Aquac. Res. 2010, 41, 1691–1698. [Google Scholar] [CrossRef]
  42. Wang, W.J.; Sun, D.Y.; Pan, B.H.; Sun, X.F. Effects of Bacillus subtilis added in feed on growth, immunity and specific intestinal microflora of grass carp. Feed. Res. 2014, 19, 43–44. [Google Scholar]
  43. Qin, L.; Xiang, J.; Xiong, F.; Wang, G.; Zou, H.; Li, W.; Li, M.; Wu, S. Effects of Bacillus licheniformis on the growth, antioxidant capacity, intestinal barrier and disease resistance of grass carp (Ctenopharyngodon idella). Fish Shellfish. Immunol. 2020, 97, 344–350. [Google Scholar] [CrossRef]
  44. Sun, N.; Wang, T.; Han, H.; Zhang, M.; Wang, F.; Jiang, H. Isolation, identification and characteristics of two Bacillus associated from intestine of marine fishes. Mar. Fish. 2019, 41, 606–615. [Google Scholar]
  45. Lutfi, E.; Basili, D.; Falcinelli, S.; Morillas, L.; Carnevali, O.; Capilla, E.; Navarro, I. The probiotic Lactobacillus rhamnosus mimics the dark-driven regulation of appetite markers and melatonin receptors’ expression in zebrafish (Danio rerio) larvae: Understanding the role of the gut microbiome. Comp. Biochem. Physiol. Part B Biochem. Mol. Biol. 2021, 256, 110634. [Google Scholar] [CrossRef]
  46. Yin, Z.; Liu, Q.; Liu, Y.; Gao, S.; He, Y.; Yao, C.; Ai, Q. Early life intervention using probiotic Clostridium butyricum improves intestinal development, immune response, and gut microbiota in large yellow croaker (Larimichthys crocea) larvae. Front. Immunol. 2021, 12, 640767. [Google Scholar] [CrossRef]
  47. Biedermann, L.; Rogler, G. The intestinal microbiota: Its role in health and disease. Eur. J. Pediatr. 2015, 174, 151–167. [Google Scholar] [CrossRef] [PubMed]
  48. Chen, M.; Liu, C.; Dai, M.Z.; Wang, Q.; Li, C.; Hung, W. Bifidobacterium lactis Bacillus licheniformis-99 modulates intestinal inflammation and functions in zebrafish models. PLoS ONE 2022, 17, e0262942. [Google Scholar]
  49. Xiao, F.; Zhu, W.; Yu, Y.; Huang, J.; Li, J.; He, Z.; Yan, Q. Interactions and stability of gut microbiota in zebrafish increase with host development. Microbiol. Spectr. 2022, 10, e0169621. [Google Scholar] [CrossRef] [PubMed]
  50. Hu, C.; Huang, Z.; Sun, B.; Liu, M.; Tang, L.; Chen, L. Metabolomic profiles in zebrafish larvae following probiotic and perfluorobutanesulfonate coexposure. Environ. Res. 2022, 204, 112380. [Google Scholar] [CrossRef]
  51. He, Y.; Guo, X.; Tan, B.; Dong, X.; Yang, Q.; Liu, H.; Chi, S. Replacing fish meal with fermented rice protein in diets for hybrid groupers (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂): Effects on growth, digestive and absorption capacities, inflammatory related gene expression, and intestinal microbiota. Aquac. Rep. 2021, 19, 100603. [Google Scholar] [CrossRef]
  52. Wu, Z.X.; Feng, X.; Qu, Y.; Chen, X.X. Effects of Bacillus subtilis on the growth, digestion and intestinal flora of grass carp. Abstract Collection of Papers from the 9th World Chinese Fish and Shrimp Nutrition Academic Symposium, Xiamen, China, 12–16 November 2013; Volume 366, p. 383. [Google Scholar]
Fishes 10 00189 g001
Figure 1. Histogram of the intestinal microbial genus distribution. Note: A: without Bacillus subtilis; B, C, D: Bacillus subtilis supplementation levels of 0.10%, 0.20% and 0.40%, respectively. Group A served as the control without B. licheniformis supplementation, while experimental groups B, C and D received graded concentrations of B. licheniformis at 0.10%, 0.20% and 0.40% (w/w), respectively.
Figure 1. Histogram of the intestinal microbial genus distribution. Note: A: without Bacillus subtilis; B, C, D: Bacillus subtilis supplementation levels of 0.10%, 0.20% and 0.40%, respectively. Group A served as the control without B. licheniformis supplementation, while experimental groups B, C and D received graded concentrations of B. licheniformis at 0.10%, 0.20% and 0.40% (w/w), respectively.
Fishes 10 00189 g001
Table 1. Composition and nutrient levels of the basal diet (in DM, %).
Table 1. Composition and nutrient levels of the basal diet (in DM, %).
ItemsContent
IngredientsGroup AGroup BGroup CGroup D
Casein28.00 28.00 28.00 28.00
Gelatin10.00 10.00 10.00 10.00
Fish meal8.00 8.00 8.00 8.00
Soybean meal8.00 8.00 8.00 8.00
Fish oil7.00 7.00 7.00 7.00
Soybean oil7.00 7.00 7.00 7.00
Dextrin21.50 21.40 21.30 21.10
Choline0.20 0.20 0.20 0.20
Ca(H2PO4)21.00 1.00 1.00 1.00
Premix a2.50 2.50 2.50 2.50
Cellulose6.80 6.80 6.80 6.80
probiotics0.00 0.10 0.20 0.40
Total100.00 100.00 100.00 100.00
Nutrients level b
Crude protein43.61 43.60 43.60 43.60
Crude lipid12.11 12.10 12.10 12.10
(MJ·kg−1) Gross energy19.49 19.48 19.41 19.39
Lysine3.03 3.03 3.03 3.03
Methionine1.02 1.02 1.02 1.02
Note: a. The premix contained the following components per kg of feed: 55 mg vitamin B1, 215 mg vitamin B2, 45 mg vitamin B6, 25 mg vitamin B12, 20 mg folic acid, 15 mg vitamin C3 (30%), 380 mg pantothenic acid, 500 mg inositol, 5 mg D-biotin (2%), 700 mg niacin, 25 mg vitamin A, 100 mg vitamin E (50%), 35 mg vitamin D, 20 mg vitamin K, 850 mg Ca(H2PO4)2·H2O, 340 mg KH2PO4, 510 mg NaCl, 800 mg MgSO4·7H2O, 660 mg NaH2PO4·2H2O, 1.5 mg KI, 15 mg CuSO4·5H2O, 340 mg ZnSO4·7H2O, 260 mg FeSO4·7H2O, 70 mg MnSO4·4H2O, 5 mg Na2SeO3, 3.5 mg CoSO4·6H2O. b. The nutrient levels are calculated values. Group A served as the control without B. licheniformis supplementation, while experimental groups B, C and D received graded concentrations of B. licheniformis at 0.10%, 0.20% and 0.40% (w/w), respectively.
Table 2. Effects of B. licheniformis on the growth performance and body indices of hybrid sturgeon (n = 10).
Table 2. Effects of B. licheniformis on the growth performance and body indices of hybrid sturgeon (n = 10).
ItemsGroup AGroup BGroup CGroup Dp Value
% SR92.50 ± 2.0195.00 ± 2.1197.50 ± 2.2297.50 ± 2.040.847
g IBW850.10 ± 12.37851.31 ± 11.22853.24 ± 11.35853.44 ± 13.110.782
g FBW1442.33 ± 14.77 a1468.45 ± 15.64 a1552.11 ± 15.41 b1502.12 ± 14.50 ab0.045
% WG69.67 ± 3.34 a72.49 ± 2.85 a81.90 ± 2.88 b76.01± 2.93 ab0.032
SGR
(%·d−1)		0.44 ± 0.02 a0.45 ± 0.02 a0.50 ± 0.02 b0.47 ± 0.01 ab0.022
FCR1.88 ± 0.03 a1.55 ± 0.04 b1.46 ± 0.05 b1.50 ± 0.04 b0.018
% VSI13.75 ± 0.4313.40 ± 0.2613.02 ± 0.2913.11 ± 0.300.523
% HSl8.88 ± 0.158.63 ± 0.148.22 ± 0.188.33 ± 0.160.634
Note: Values in the same row with no letter or the same superscripted letter had no significant difference (p > 0.05), whereas those with different superscripted letters had significant differences (p < 0.05). Group A served as the control without B. licheniformis supplementation, while experimental groups B, C and D received graded concentrations of B. licheniformis at 0.10%, 0.20% and 0.40% (w/w), respectively.
Table 3. Effect of B. licheniformis on the body composition of sturgeon (n = 10).
Table 3. Effect of B. licheniformis on the body composition of sturgeon (n = 10).
ItemsGroup AGroup BGroup CGroup Dp Value
CP18.44 ± 0.64 a19.95 ± 0.61 b20.21 ± 0.62 b20.11 ± 0.57 b0.032
EE5.58 ± 0.51 a6.61 ± 0.52 b6.96 ± 0.55 b6.87 ± 0.47 b0.044
Ash 3.33 ± 0.373.45 ± 0.443.61 ± 0.413.57 ± 0.400.511
MS79.67 ± 0.2580.19 ± 0.2180.44 ± 0.2380.32 ± 0.250.722
Note: Values in the same rows with no letter or the same superscripted letter had no significant difference (p > 0.05), whereas those with different superscripted letters had significant differences (p < 0.05). CP is crude protein. EE is ether extract. MS is water content. The content of crude protein (CP) was determined by Kjeldahl method (GB 5009.5-2016, [21]), the content of crude fat (EE) was determined by Soxhlet extractor method (GB 5009.6-2016, [22]), the ash was determined by Muffle furnace ashing method (GB 5009.4-2016, [23]) and the water content (MS) was Direct drying method (GB 5009.3-2016, [24]). Group A served as the control without B. licheniformis supplementation, while experimental groups B, C and D received graded concentrations of B. licheniformis at 0.10%, 0.20% and 0.40% (w/w), respectively.
Table 4. Effects of B. licheniformis on the plasma biochemical parameters of sturgeon (n = 3).
Table 4. Effects of B. licheniformis on the plasma biochemical parameters of sturgeon (n = 3).
ItemsGroup AGroup BGroup CGroup Dp Value
TP (g/L)19.11 ± 0.2219.43 ± 0.2219.75 ± 0.2119.61 ± 0.150.931
TG (mmol/L)7.70 ± 0.23 a8.93 ± 0.25 b9.22 ± 0.21 b9.11 ± 0.24 b0.021
TC (mmol/L)2.42 ± 0.11 a3.55 ± 0.13 b3.87 ± 0.14 b3.41 ± 0.15 b0.029
ALP (U/L)151.13 ± 7.12154.22 ± 7.03158.41 ± 6.80156.10 ± 7.400.755
ALT (U/L)7.02 ± 0.68 a7.85 ± 0.54 ab8.07 ± 0.51 b8.00 ± 0.54 b0.041
AST (U/L)556.41 ± 23.11 a600.23 ± 24.13 ab677.81 ± 25.96 b660.18 ± 26.22 b0.038
GLU (mmol/L)2.50 ± 0.14 a3.11 ± 0.15 b3.74 ± 0.13 ab2.94 ± 0.12 ab0.026
Note: In the same row, values with no letter or the same superscripted letter are not significantly different (p > 0.05), whereas those with different superscripted letters are significantly different (p < 0.05). Group A served as the control without B. licheniformis supplementation, while experimental groups B, C and D received graded concentrations of B. licheniformis at 0.10%, 0.20% and 0.40% (w/w), respectively.
Table 5. Diversity indices of the intestinal microbiota samples.
Table 5. Diversity indices of the intestinal microbiota samples.
ItemsGroup AGroup BGroup CGroup Dp Value
OTU265.11 ± 7.61 a211.01 ± 8.42 a185.44 ± 8.32 ab143.24 ± 7.61 b0.011
ACE277.19 ± 3.15 a265.40 ± 2.41 a211.42 ± 2.11 ab175.10 ± 2.32 b0.045
Chao1188.11 ± 4.12 a205.44 ± 3.16 ab279.30 ± 3.02 b231.22 ± 3.22 b0.033
Simpson0.68 ± 0.02 a0.81 ± 0.02 b0.87 ± 0.02 b0.84 ± 0.01 b0.022
Shannon0.48 ± 0.03 a0.65 ± 0.03 b0.79 ± 0.14 b0.68 ± 0.05 b0.041
Note: In the same row, values with no letter or the same superscripted letter are not significantly different (p > 0.05), whereas those with different superscripted letters are significantly different (p < 0.05). Group A served as the control without B. licheniformis supplementation, while experimental groups B, C and D received graded concentrations of B. licheniformis at 0.10%, 0.20% and 0.40% (w/w), respectively.
Table 6. Community composition of intestinal microbial at the genus level.
Table 6. Community composition of intestinal microbial at the genus level.
Gut MicrobeGroup AGroup BGroup CGroup D
Cetobacterium91.2289.2387.6583.22
Plesiomonas3.014.165.029.21
Streptococcus2.021.551.331.01
Clostridium0.10.190.551.1
Lactococcus0.850.570.520.42
Lactobacillus0.891.021.781.98
Bacillus0.011.752.00 2.12
Candidatus Competibacter0.550.310.270.19
Sphingomonas1.351.220.880.75
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Xie, Q.; Wang, Y.; Huang, X.; Liu, Y.; Yang, M.; Xing, H.; Yang, C.; Hu, C.; Pan, M.; Qi, Z. Effects of Bacillus licheniformis Feeding on the Growth Performance, Blood Parameters and Intestinal Microbiota of Adult Hybrid Sturgeon. Fishes 2025, 10, 189. https://doi.org/10.3390/fishes10050189

AMA Style

Xie Q, Wang Y, Huang X, Liu Y, Yang M, Xing H, Yang C, Hu C, Pan M, Qi Z. Effects of Bacillus licheniformis Feeding on the Growth Performance, Blood Parameters and Intestinal Microbiota of Adult Hybrid Sturgeon. Fishes. 2025; 10(5):189. https://doi.org/10.3390/fishes10050189

Chicago/Turabian Style

Xie, Quansen, Yu Wang, Xinyu Huang, Yiran Liu, Mingjian Yang, Haochun Xing, Caimei Yang, Caihong Hu, Mingzhu Pan, and Zhitao Qi. 2025. "Effects of Bacillus licheniformis Feeding on the Growth Performance, Blood Parameters and Intestinal Microbiota of Adult Hybrid Sturgeon" Fishes 10, no. 5: 189. https://doi.org/10.3390/fishes10050189

APA Style

Xie, Q., Wang, Y., Huang, X., Liu, Y., Yang, M., Xing, H., Yang, C., Hu, C., Pan, M., & Qi, Z. (2025). Effects of Bacillus licheniformis Feeding on the Growth Performance, Blood Parameters and Intestinal Microbiota of Adult Hybrid Sturgeon. Fishes, 10(5), 189. https://doi.org/10.3390/fishes10050189

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