Synergistic Effect of Dietary Inactivated <i>Lactobacillus plantarum</i> and Berberine Supplementation on Growth Performance, Antioxidant Capacity, and Immune Function of Juvenile Black Sea Bream (<i>Acanthopagrus schlegelii</i>)

Sagada, Gladstone; Wang, Lei; Xu, Bingying; Tegomo, Fabrice Arnaud; Chen, Kai; Zheng, Lu; Sun, Yuxiao; Liu, Yuechong; Yang, Yifei; Ullah, Sami; Shao, Qingjun

doi:https://doi.org/10.1155/2022/3053724

Aquaculture Nutrition

On this page

Abstract Introduction Materials and Methods Results Discussion Conclusions Data Availability Conflicts of Interest Acknowledgments References Copyright Related Articles

Research Article | Open Access

Volume 2022 | Article ID 3053724 | https://doi.org/10.1155/2022/3053724

Synergistic Effect of Dietary Inactivated Lactobacillus plantarum and Berberine Supplementation on Growth Performance, Antioxidant Capacity, and Immune Function of Juvenile Black Sea Bream (Acanthopagrus schlegelii)

Gladstone Sagada,¹Lei Wang,^1,2Bingying Xu,¹Fabrice Arnaud Tegomo,^3,4Kai Chen,¹Lu Zheng,¹Yuxiao Sun,¹Yuechong Liu,¹Yifei Yang,⁵Sami Ullah,¹and Qingjun Shao^1,3

Academic Editor: Mahmoud Dawood

Received29 Dec 2021

Revised15 Feb 2022

Accepted17 Mar 2022

Published29 Mar 2022

Abstract

The interactive effect of dietary inactivated Lactobacillus plantarum and berberine on black sea bream, Acanthopagrus schlegelii, was investigated with three diets designated as D1 (Con: basal diet), D2 (LP: basal diet + 400 mg/kg L. plantarum), and D3 (LPBB: basal diet + 400 mg/kg L. plantarum + 50 mg/kg berberine) and fed to juvenile black sea bream ( g) for 56 days. The growth performance and feed utilization parameters, as well as intestinal trypsin activity, were significantly improved in the LP and LPBB groups (). Fish fed the LPBB diet showed better serum and hepatic antioxidant capacity, whilst the LP group had better hepatic antioxidant capacity, than the control fish (). Intestinal IgM and C3 levels significantly increased in the LPBB fish than the rest of the groups (). NF-κB was significantly upregulated in the LP group (). Nrf2 and IL-10 were significantly upregulated, whilst Keap1b and NF-κB were significantly downregulated in the LPBB group (). These findings show that a combination of inactivated L. plantarum and berberine in the diet of black sea bream can improve the immune response and antioxidant capacity than a diet with only the inactivated probiotic, whilst both diets can equally improve growth performance.

1. Introduction

Global aquaculture production has been growing consistently over the past decades than all the other food sectors partly due to continuous research and innovation [1]. The use of functional feed additives to alleviate losses to stress, pests, and disease outbreak has been an essential research focus. With the use of antibiotics becoming less accepted due to the detrimental effects of their abuse on aquatic environments, animal natural immune function, and risk of consumers developing super bugs [2, 3], there has been copious research into alternative functional additives such as probiotics [4, 5], plant extracts [6, 7], prebiotics [8, 9], and organic acids [10, 11], which are considered more environmentally friendly and less likely to develop resistant bacteria.

Berberine is an organic compound present in herbs like Coptis chinensis, Berberis thunbergii, and Hydrastis canadensis, with huge popularity in traditional Chinese medicine. This isoquinoline alkaloid has a long history dating back to ancient China in its use against proliferation, inflammation, and hypertension [12, 13]. Natural materials like hydrochloride present in berberine give it the ability to ameliorate the impairment of hepatocyte function and excessive lipid in the blood [14]. The potential of berberine to help improve productivity in aquaculture through its prophylactic effects has been demonstrated in different fish species, including grass carp, Ctenopharyngodon idella [15]; blunt snout bream, Megalobrama amblycephala [16]; and black sea bream, Acanthopagrus schlegelii [17].

According to the FAO/WHO, probiotics are live microorganisms that have beneficial effects on the host when administered in ample amount [18]. Although probiotics are expected to be viable in order to perform their functions, there is evidence of “dead” probiotics and their metabolites eliciting similar responses as their live forms albeit with potential differences [19, 20]. Inactivated probiotics are considered a safer alternative to live forms owing to the risk of live microorganisms distorting the ecosystem of the rearing environs, especially in open waters. Furthermore, inactivated bacteria are easier to transport and store [21], and can withstand the harsh conditions associated with feed processing than live forms, making them more suited for aquafeed. Lactobacillus plantarum belongs to a group of microorganisms called lactic acid bacteria (LAB), which generate lactic acid as the main product of carbohydrate fermentation, with a wide range of application in nutrition [22, 23]. Inactivated L. plantarum was shown to have the potential to improve growth performance and/or health condition in different fish species, including red sea bream, Pagrus major [24]; Nile tilapia, Oreochromis niloticus [25]; bighead catfish, Clarias macrocephalus [26]; snakehead, Channa striata [27]; and black sea bream [28].

Black sea bream is a very important commercial fish species in many parts of Asia and is widely distributed along the Northwest Pacific, from the coasts of Japan and Korea to the South China Sea [29]. The commercial and recreational importance of this species has led to depletion of wild stocks due to pressure from overfishing [30]. Aquaculture production of black sea bream is therefore essential to reduce pressure on the wild stocks. Several studies have investigated the effect of combining dietary probiotics with other functional additives on different fish species: L. plantarum was combined with beta-glucan in diets of red sea bream [31]; Bacillus subtilis was combined with mannanoligosaccharide for Japanese eel, Anguilla japonica [32]; and the interactive effect between L. plantarum and pineapple peel powder was investigated on Nile tilapia [33]. Drawing inference from the earlier reports that explored the possible synergies between dietary probiotics and other functional additives and previous studies in our laboratory, which separately demonstrated positive effects of inactivated L. plantarum (), and berberine (50 mg/kg) on black sea bream [17, 28], this study further explored the possible interactive effects of L. plantarum and berberine on growth performance, antioxidant capacity, and immune response of this species. With no previous studies on a combination between the two additives in the diet of any farmed animal to the best of our knowledge, the study focused on whether adding berberine to L. plantarum could achieve a better impact than the probiotic-only diet. This will add to the alternatives for producing functional feeds that can boost productivity in black sea bream farming.

2. Materials and Methods

2.1. Diet Preparation

The inactivated probiotic (Immuno-LP20) is a product of House Wellness Foods Corp. (Itami, Japan). The procedure for preparation was as described previously [34]. It is an off-white powder with cfu g^-1 inactivated L. plantarum (L-137) (20%) and dextrin hydrolysed from tapioca (80%). Berberine () was procured from Spring and Autumn Biotechnology Company (Nanjing, China). The basal diet was formulated to contain the same crude protein and crude lipid levels (Table 1). The main protein ingredients (fishmeal, fermented soybean meal, and soybean protein concentrate) were finely ground and sieved with a 180 μm mesh, and samples were taken from each ingredient for proximate composition analysis. Fish oil, soy lecithin, and corn oil were used as the main lipid sources. The diets were designated as D1 (Con: basal diet, control), D2 (LP: basal diet + 400 mg/kg L. plantarum), and D3 (LPBB: basal diet + 400 mg/kg L. plantarum + 50 mg/k berberine). Methionine and lysine were supplemented, and the basal diet was formulated based on earlier reports on black sea bream [35, 36], whilst the L. plantarum and berberine inclusion levels were based on our previous studies [17, 28]. The ingredients were mixed and homogenized to form a dough, which was extruded via a 2.5 mm mould feed machine (HKJ-218, HUARUI, Wuxi, China). The pellets were then steamed for about 10 minutes and dried at 24°C for about 72 hours in a ventilated room and stored at -20°C. Representative samples were used for proximate composition analysis.

2.2. Experimental Setup and Feeding

The experimental procedures in this study were according to the Guidelines of the Care and Use of Laboratory Animals in China, and the study was approved by the Committee on the Ethics of Animal Experiments of Zhejiang University (Ethics code: ZJU20200076). All fish were handled with care during all the stages of this experiment.

Black sea bream fingerlings were obtained from a hatchery close to the rearing facility and acclimated with a commercial feed (42% crude protein, Ming Hui Co. Ltd.) for a 14-day period. The rearing experiment took place at the Zhejiang University Test Base for Marine Science and Ocean Engineering located on Zhai Ruo Shan Island, East China Sea (Zhoushan, China). After a successful acclimation, the fish were starved for 24 hours, and healthy ones in good physical condition without ragged fins, swollen abdomen, hemorrhages, and exophthalmia were selected () and stocked in 9 cylindrical tanks. Each tank had 40 fish in 700 L seawater, and they were randomly divided into triplicates of three treatments. The tanks were connected to a flow-through system with continuous supply of sea water filtered with biofilter and drum filter (2 L/min flow rate). The tanks were continuously aerated with air stones for optimum dissolved oxygen (7.1–9.5 ppm). Temperature (26.5–28.2°C), pH (8.1–8.3), and salinity (27–29 g/L) were also monitored every day. The fish were fed to satiation twice daily (8:00 and 15:00) during the eight-week rearing period. Each tank was observed regularly to remove dead fish.

2.3. Sampling and Biochemical Analysis

After the feeding period, the fish were anesthetized with 60 mg/L MS-222 after a 24-hour starvation. Thereafter, they were individually counted and measured for weight and length. Blood was drained from the caudal vein with 27-gauge needle and 1 mL syringe, allowed to settle for 2 hours and centrifuged at 10,000 g for 20 minutes (4°C) to obtain the serum. The liver, viscera, and intraperitoneal fat were weighed and recorded after dissection to determine body condition indices, and dorsal muscle, foregut, and liver samples were stored at -80°C with whole body and serum for proximate analysis and assay. The intestine samples were also frozen in liquid nitrogen for gene analysis.

Proximate analysis of fish and feed samples were conducted using the methods of the Association of Official Analytical Chemists [37]. Supernatants of the intestine and liver samples for assays were separated following the process used in Sagada et al. [38]. The diagnostic reagent kits for chemical analyses were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Glutathione S-transferase (GST) activity was assayed as described by Elia et al. [39]. Malondialdehyde (MDA) content and catalase (CAT), glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) activities were determined as described by Han et al. [40]. Concentrations of immunoglobulin (IgM) and complement (C3 and C4) proteins were analysed using the enzyme-linked immunosorbent assay (ELISA) method [41]. Intestinal digestive enzymes (trypsin, amylase, and lipase) were determined according to the assay protocols described by Li et al. [42].

2.4. Quantitative PCR Assay

Total RNA extracted from four replicates of each intestine sample (TaKaRa MiniBEST Universal RNA Extraction Kit, Takara, Japan) were used for reverse transcription with PrimeScript™ RT reagent Kit with gDNA Eraser (Perfect Real Time, Takara, Japan). The primer pairs for the various genes were designed by Primer Premier 5.0 using RNAseq data previously obtained in our lab (Table 2). SYBR Premix Ex Taq™ II Kit (Takara, Japan) was used for the real-time qPCR with the following process: 95°C for 30 s, 40 cycles of 95°C for 5 s, followed by 60°C for 30 s. β-Actin was used as the reference gene, and the fold change was calculated with the 2^−ΔΔCT method [43].

2.5. Statistical Analysis

All the data are shown as the (SD) (). SPSS 20.0 (IBM SPSS, Chicago, IL, USA) was used to conduct one-way analysis of variance (ANOVA), which tested the effects of the diets on the various parameters. Tukey’s multiple range test was used to check for differences between the means (), normality was determined with the Kolmogorov–Smirnov test and homogeneity of variance checked by Levene’s test.

3. Results

3.1. Growth, Body Condition, and Feed Utilization

According to the growth parameters shown in Table 3, fish fed the diets with inactivated L. plantarum-only (LP), and a combination of the inactivated probiotic and berberine (LPBB) had significantly higher final body weight (FBW), weight gain (WG), and specific growth rate (SGR) than the control group (), and the growth parameters did not differ significantly between both of the supplemented groups (). The body condition indicators (viscerasomatic index, condition factor, hepatosomatic index, and intraperitoneal fat ratio) and survival rate were not significantly affected by the dietary treatments (). The impacts of the diets on feeding rate (FR), protein productive value (PPV), protein efficiency ratio (PER), daily feed intake per fish (DFI), total feed intake (TFI), and feed conversion ratio (FCR) are shown in Table 4. FR, PPV, and PER were not significantly affected by the dietary treatments (). DFI and TFI significantly increased, whilst FCR significantly decreased in fish fed LP and LPBB in comparison with the control fish (). There was however no significant difference in the feed intake and utilization parameters between the LP and LPBB groups ().

3.2. Proximate Body Composition

The muscle and whole-body moisture, crude protein, ash, and crude lipid contents of fish fed the test diets were not significantly impacted () as shown in Table 5.

3.3. Serum and Liver Antioxidant Parameters

As shown in Table 6, hepatic GSH-Px activity, serum CAT and GST activities, and serum MDA content were not significantly impacted by the dietary treatments (). Fish fed LPBB had significantly higher Serum SOD activity than the control fish and significantly higher GSH-Px activity than the rest of the groups (). The hepatic SOD activity was significantly higher in fish fed LP than the control group, and the GST activity significantly higher than the rest of the groups (). There was significantly higher liver CAT activity in the LP and LPBB groups than fish fed the control diet ().

3.4. Intestinal Digestive Enzyme Activity and Immune Parameters

The activities of intestinal lipase (Figure 1(b)) and amylase (Figure 1(c)) in black sea bream were not significantly impacted by the dietary treatments (), but trypsin activity (Figure 1(a)) was significantly higher in the LP and LPBB groups than the control fish (). C4 content (Figure 2(c)) of the test fish was not significantly affected by the dietary treatments (), but the IgM and C3 protein contents (Figures 2(a) and 2(b)) were significantly higher in fish fed the LPBB diet than the rest of the groups ().

(a)

(b)

(c)

(a)

(b)

(c)

3.5. Intestinal Gene Expression

The relative expression levels of interferon γ (IFN-γ) (Figure 3(a)), interleukin-10 (IL-10) (Figure 3(b)), nuclear factor-kappa B (NF-κB) (Figure 3(c)), nuclear factor erythroid-2-related factor 2 (Nrf2) (Figure 3(d)), and Kelch-like ECH-associated protein 1b (Keap1b) (Figure 3(e)) of black sea bream were assessed. IFN-γ was not significantly affected by the dietary treatments (). The LP fish had significantly upregulated NF-κB than the rest of the groups and significantly downregulated Keap1b than the control fish (). The LPBB fish had significantly upregulated Nrf2 and IL-10 and significantly downregulated Keap1b and NF-κB than the rest of the groups ().

(a)

(b)

(c)

(d)

(e)

Figure 3

Relative expression levels of immune and antioxidant-related genes in foregut of black sea bream fed the experimental diets. Different letters on bars show significant difference (). (a) IFN-γ: Interferon γ; (b) IL-10: interleukin-10; (c) NF-κB: nuclear factor-kappa B; (d) Nrf2: nuclear factor erythroid-2-related factor 2; (d) Keap1b: Kelch-like ECH-associated protein 1b. Con, fish fed D1: basal diet; LP, fish fed D2: basal diet+400 mg/kg L. plantarum; LPBB, fish fed D3: basal diet+400 mg/kg L. plantarum+50 mg/kg berberine.

4. Discussion

The interactive effect of dietary probiotics in combination with other nonantibiotic functional additives is being investigated in recent times in the quest to develop functional feeds to improve production of different aquaculture species. This study sets out to compare the effects of dietary inactivated L. plantarum alone (LP) with that of a combination between L. plantarum and berberine (BB) on black sea bream. No berberine-only treatment was included in this trial because previous studies in our laboratory showed how berberine supplementation in black sea bream diets improved the health condition, evinced in antioxidant function, inflammatory cytokines, and other health-related parameters [17, 44, 45]. The FBW, WG, and SGR results from the study show that the combination (LPBB) significantly improved growth performance, but the synergistic effect of the combination could not significantly improve the growth than the probiotic alone, despite the slightly higher feed intake and feed utilization it produced. The higher growth in fish fed both LP and LPBB can be attributed to better appetite and higher feed intake due to the significantly less feed eaten by the control fish. The ability of dietary probiotics to improve growth by increasing nutrient availability and utilization through synthesizing extracellular enzymes, fatty acids, vitamins, and amino acids [46, 47] was shown in the growth and FCR values of this study. Similar to these results, previous reports showed growth-promoting effects of inactivated probiotics on different fish species, including grouper by Bacillus pumilus [48], tilapia by L. plantarum [49], red sea bream by L. plantarum [24], and black sea bream by L. plantarum [28]. Berberine supplementation did not improve the growth of black sea bream in our previous study [17], and combining L. plantarum with berberine produced only slightly higher growth than the LP diet in the current result. However, berberine supplementation promoted growth and feed utilization in other fish [12, 50, 51], possibly through modulation of intestinal microbiota [52, 53]. In terms of interactive effect, this study is similar to an earlier one in which dietary Lactobacillus rhamnosus GG combined with Jerusalem artichoke, Helianthus tuberosus, improved the growth performance of red tilapia, Oreochromis spp., than the nonsupplemented control diet, but there was no significant difference when compared with fish fed the diet with only the probiotic [54]; and this contradicts another study in which dietary L. plantarum+Jerusalem artichoke significantly improved growth performance than both the control and probiotic-only diets administered to Pangasius catfish, Pangasius bocourti [55]. The current result is also dissimilar to our earlier study, in which a combination of selenomethionine and inactivated L. plantarum caused a decrease in growth of black sea bream [56]. The inability of the combination of both L. plantarum and berberine to improve growth than the LP diet could be because similar to berberine, dietary probiotics can also improve the growth of fish by modulating the intestinal microbiota [57, 58]. The similarity in their growth-promoting mechanisms could have inhibited additivity of their separate effects since they both could have conferred the same benefit in this regard. Considering the additivity of the impacts of various feed supplements could help establish the possibility of their synergy.

Oxidation and antioxidant parameters were measured vis-à-vis the dietary treatments to ascertain the ability of the various diets to provide protection against potential oxidative damage. Hepatic SOD, CAT, and GST activities, as well as serum GSH-Px and SOD activities, were significantly impacted. Interestingly, only the LPBB diet caused significantly increased serum antioxidant enzyme activities than the control fish, but both the LP and LPBB diets conferred significantly higher antioxidative protection on the liver. The improved antioxidant parameters in both the liver and serum of the LPBB fish compared to improvement in only the liver in the LP group could be an indication of better protection by the combination of L. plantarum and berberine against oxidative stress than the probiotic alone, but it is not clear why the LP diet could not improve the parameters in the serum as well. In line with this result, dietary Pseudomonas aerugenosa VSG2 and L. plantarum VSG3 could not significantly affect the serum SOD activity in rohu, Labeo rohita [59, 60]. The antioxidant enzymes GSH-Px, SOD, CAT, and GST all provide antioxidative defence via different mechanisms. The antioxidant defence system of fish works to prevent the buildup of reactive oxygen species (ROS) such as superoxides, hydroxyl radicals, and peroxides, which cause oxidative stress [61]. Comparable to the current results, Bacillus supplementation in diets boosted the antioxidant capacity of grass carp, Ctenopharyngodon idellus [62, 63]; dietary berberine enhanced the antioxidant defence system in blunt snout bream [50] and black sea bream [44]; whilst B. licheniformis and fructooligosaccharide supplemented together in a diet improved the antioxidant defence ability of triangular bream, Megalobrama terminalis [64].

Probiotics are believed to boost the antioxidant defence system by enhancing the secretion of antioxidant enzymes [63, 64], whilst berberine was shown to attenuate oxidative stress through improving mitochondria density and increasing activities of complex I and complex II by upregulating SirT3 expression [65], or via the p38 mitogen-activated protein kinase (MAPK) pathway [66]. Furthermore, probiotics and berberine can both regulate antioxidant capacity through the Nrf2/Keap1 signaling pathway. Nrf2 is a transcription factor, which binds with antioxidant response element (ARE) sequences to produce antioxidant enzymes, and Keap1 is a molecular switch that holds Nrf2 from translocating into the nucleus to bind with ARE [67]. L. plantarum modulated the Nrf2-Keap1-ARE pathway in mice [68], whilst berberine was also shown to activate the Nrf2 antioxidant pathway in rats [69]. In this study, the LPBB diet significantly upregulated Nrf2 and downregulated Keap1b in the intestine of black sea bream than the rest of the diets, and the LP diet significantly downregulated Keap1b than the control diet. This indicates the tendency of a diet with both L. plantarum and berberine to modulate the Nrf2/Keap1 signaling pathway than L. plantarum alone. Interestingly, this bears similarity to the antioxidant enzyme activity in this study, since fish fed the LPBB diet had significantly higher antioxidant indices in both the serum and liver, whilst the LP fish only had significantly higher hepatic antioxidant parameters, than the control fish. From the results of the antioxidant indices, as well as the expression of Nrf2 and Keap1b genes, it can be posited that supplementing both berberine and L. plantarum in the diet of black sea bream could confer better protection against oxidative stress than the inactivated probiotic alone.

The tendency of probiotics to increase digestive enzyme activity possibly through secretion of extracellular enzymes [70] has been well documented in different fish species: B. subtilis supplementation in diets of Nile tilapia increased protease and amylase activities [71]; B. subtilis and Lactococcus lactis improved trypsin activity in Nile tilapia [72]; and Enterococcus faecalis inclusion in diets increased protease and lipase activities in Javanese carp, Puntius gonionotus [73]. The ability of berberine to modulate intestinal microbiota could somewhat influence digestive enzymes, but there is paucity of empirical evidence to corroborate that. The significantly higher trypsin activity in fish fed both the LP and LPBB diets correlates with the growth and feed utilization results and can be attributed to enzyme-secreting ability of L. plantarum and the influence of the probiotic and berberine on intestinal microbiota and antioxidative status, which are beneficial to intestinal integrity and digestive enzymes. Continuous exposure to probiotics could inhibit endogenous enzyme activity [58] and may have affected lipase and amylase activities in this study.

The interaction between ingested feed and the enteric sensory neurons highly influences intestinal health, which encompasses immune function, structure, and microbiome. The intestine in animals is relatively large in comparison to other major immune organs; hence, it plays a vital role in helping fish resist pathogen invasion. The IgM and C3 protein contents in the foregut of black sea bream in this study were significantly increased by inclusion of inactivated L. plantarum and berberine together. The immunoglobulin and complement system are essential parts of an animal’s immune system. As the predominant antibody in fish, the IgM helps provide protection against different pathogens [64]. C3 is part of the complement cascade system in teleost fish and can cause the death of pathogens by perforating their surface membrane [74]. The higher levels of C3 and IgM in fish fed the LPBB diet than both the control and LP groups means L. plantarum and berberine could be functionally synergistic in modulating immune response in black sea bream. Similarly, inactivated L. plantarum supplementation did not significantly impact intestinal IgM and C4 in our previous study on the same fish species [28] and is attributable to immunity fatigue from the probiotic supplementation [60, 75], which may have been ameliorated by berberine in this study. The immunostimulant effect of berberine in this study is similar to earlier reports on blunt snout bream [12, 50], and Nile tilapia [51]. The immune-stimulating ability of berberine is attributable to its influence on enteric glial cells (EGC), bone marrow-derived dendritic glial cells (BMDCs), T cells, and gut epithelial cells (IEC) [76]. Another mechanism by which berberine modulates immune response could be by attenuating the TLR4/NF-κB-p65 signaling pathway and activating the adenosine monophosphate- (AMP-) activated protein kinase (AMPK) signaling [77].

As a first line of defense against pathogens, IECs crucially sense signals from different sources of stimuli, including beneficial bacteria, which it in turn uses to produce cytokines and chemokines for maintenance of gut homeostasis [78, 79]. The integrity of IECs can be compromised by proinflammatory cytokines, leading to disorders, and the anti-inflammatory cytokines serve to counteract this effect. IFN-γ, a proinflammatory cytokine, was not significantly affected by the dietary treatments in this study. Fish fed the LPBB diet had significantly downregulated NF-κB and upregulated IL-10, an anti-inflammatory cytokine than the rest of the groups, an indication of the ability of the combination of berberine and L. plantarum to confer higher anti-inflammatory protection on black sea bream than the probiotic alone. Furthermore, IL-10 improves immune function by aiding in differentiating immune cells like natural killer (NK) cells, dendritic cells, B cell, and granulocytes [80]. This is in line with the intestinal IgM and C3 contents in this study, which indicated that the LPBB diet conferred higher immunity than L. plantarum alone. NF-κB is a transcription factor for proinflammatory cytokines, and fish fed the LP diet had the highest NF-κB expression, indicating the possibility of inactivated L. plantarum to confer inflammatory response against pathogen invasion as seen in slightly higher IFN-γ expression than the control fish [81]. This is consistent with reports where Bacillus spp. [48, 82], and L. plantarum [83] increased the expression of proinflammatory cytokines. Berberine has been proven to have anti-inflammatory tendencies and can influence NF-κB pathways [84–86]. These results indicate that inactivated L. plantarum alone could increase intestinal proinflammatory response, and berberine inclusion together with L. plantarum can help ameliorate the possibility of excessive inflammation damage to EICs and maintain tissue homeostasis.

5. Conclusions

This study determined that combining berberine with inactivated L. plantarum can improve the growth, feed utilization, and intestinal trypsin activity of juvenile A. schlegelii, albeit not better than the probiotic-only diet. However, the synergy between the functional additives could have a better influence on the immune response as seen in higher intestinal IgM and C3 levels, as well as upregulated IL-10 and downregulated NF-κB. Combining the additives also had better impact on antioxidant capacity, which was shown in higher serum SOD and GSH-Px activities, as well as upregulated Nrf2 and downregulated Keap1b in the intestine.

Data Availability

All data for this study are available from the corresponding author on reasonable request.

Conflicts of Interest

No conflict of interest exists in this paper.

Acknowledgments

This work was funded by the Ministry of Science and Technology, China (Project No. 2020YFD0900801), and the Zhejiang Provincial Bureau of Science and Technology (Project No. 2015C02020). Our gratitude goes to the Zhejiang University Test Base for Marine Science and Ocean Engineering, Zhai Ruo Shan Island (Zhoushan, China), for making their rearing facility available and logistics support during the feeding trial. We would also like to thank the House Wellness Foods Corp., Itami, Japan, for sponsoring the L. plantarum product.

References

FAO and The State of World Fisheries and Aquaculture, “FAO,” Fisheries technical paper, vol. 2020, 2020.
View at: Google Scholar
P. Hernández Serrano, “Responsible use of antibiotics in aquaculture,” Food & Agriculture Organization, vol. 469, pp. 1–97, 2005.
View at: Google Scholar
T. Defoirdt, P. Sorgeloos, and P. Bossier, “Alternatives to antibiotics for the control of bacterial disease in aquaculture,” Current Opinion in Microbiology, vol. 14, no. 3, pp. 251–258, 2011.
View at: Publisher Site | Google Scholar
S. Yilmaz, E. Yilmaz, M. A. O. Dawood, E. Ringø, E. Ahmadifar, and H. M. R. Abdel-Latif, “Probiotics, prebiotics, and synbiotics used to control vibriosis in fish: a review,” Aquaculture, vol. 547, p. 737514, 2022.
View at: Publisher Site | Google Scholar
S. S. Giri, H. J. Kim, S. G. Kim et al., “Effects of dietary Lactiplantibacillus plantarum subsp. plantarum L7, alone or in combination with Limosilactobacillus reuteri P16, on growth, mucosal immune responses, and disease resistance of Cyprinus carpio,” Probiotics and Antimicrobial Proteins, vol. 13, no. 6, pp. 1747–1758, 2021.
View at: Publisher Site | Google Scholar
E. Ahmadifar, M. Yousefi, M. Karimi et al., “Benefits of dietary polyphenols and polyphenol-rich additives to aquatic animal health: an overview,” Reviews in Fisheries Science & Aquaculture, vol. 29, no. 4, pp. 478–511, 2021.
View at: Publisher Site | Google Scholar
M. A. O. Dawood, M. F. El Basuini, A. I. Zaineldin et al., “Antiparasitic and antibacterial functionality of essential oils: an alternative approach for sustainable aquaculture,” Pathogens, vol. 10, no. 2, p. 185, 2021.
View at: Publisher Site | Google Scholar
S. K. Song, B. R. Beck, D. Kim et al., “Prebiotics as immunostimulants in aquaculture: a review,” Fish & Shellfish Immunology, vol. 40, no. 1, pp. 40–48, 2014.
View at: Publisher Site | Google Scholar
B. Xu, G. Zhang, L. Wang, G. Sagada, J. Zhang, and Q. Shao, “The influence of dietary β-1,3-glucan on growth performance, feed utilization, antioxidative and immune status of Pacific white shrimp,Litopenaeus vannamei,” Litopenaeus vannamei, Aquaculture Nutrition, vol. 27, no. 5, pp. 1590–1601, 2021.
View at: Publisher Site | Google Scholar
S. Ullah, J. Zhang, B. Xu et al., “Effect of dietary supplementation of lauric acid on growth performance, antioxidative capacity, intestinal development and gut microbiota on black sea bream (Acanthopagrus schlegelii),” PLoS One, vol. 17, no. 1, article e0262427, 2022.
View at: Publisher Site | Google Scholar
J. A. Volatiana, G. Sagada, B. Xu, J. Zhang, W. K. Ng, and Q. Shao, “Effects of butyrate glycerides supplementation in high soybean meal diet on growth performance, intestinal morphology and antioxidative status of juvenile black sea bream,Acanthopagrus schlegelii,” Acanthopagrus schlegelii, Aquaculture Nutrition, vol. 26, no. 1, pp. 15–25, 2020.
View at: Publisher Site | Google Scholar
Q. Q. Chen, W. B. Liu, M. Zhou et al., “Effects of berberine on the growth and immune performance in response to ammonia stress and high-fat dietary in blunt snout bream Megalobrama amblycephala,” Fish & Shellfish Immunology, vol. 55, pp. 165–172, 2016.
View at: Publisher Site | Google Scholar
X. Feng, A. Sureda, S. Jafari et al., “Berberine in cardiovascular and metabolic diseases: from mechanisms to therapeutics,” Theranostics, vol. 9, no. 7, pp. 1923–1951, 2019.
View at: Publisher Site | Google Scholar
C. Huang, Y. Zhang, Z. Gong et al., “Berberine inhibits 3T3-L1 adipocyte differentiation through the PPARγ pathway,” Biochemical and Biophysical Research Communications, vol. 348, no. 2, pp. 571–578, 2006.
View at: Publisher Site | Google Scholar
H. Pan, Z. Li, J. Xie et al., “Berberine influences blood glucose via modulating the gut microbiome in grass carp,” Frontiers in Microbiology, vol. 10, p. 1066, 2019.
View at: Publisher Site | Google Scholar
C. Yu, J. Zhang, Q. Qin, J. Liu, J. Xu, and W. Xu, “Berberine improved intestinal barrier function by modulating the intestinal microbiota in blunt snout bream Megalobrama amblycephala under dietary high-fat and high-carbohydrate stress,” Fish & Shellfish Immunology, vol. 102, pp. 336–349, 2020.
View at: Publisher Site | Google Scholar
L. Wang, B. Xu, G. Sagada et al., “Dietary berberine regulates lipid metabolism in muscle and liver of black sea bream (Acanthopagrus schlegelii) fed normal or high-lipid diets,” The British Journal of Nutrition, vol. 125, no. 5, pp. 481–493, 2021.
View at: Publisher Site | Google Scholar
FAO/WHO, Joint FAO/WHO working group report on drafting guidelines for the evaluation of probiotics in food, FAO Food and Nutrition Paper, 85, London, Ontario, Canada, 2002.
C. A. Adams, “The probiotic paradox: live and dead cells are biological response modifiers,” Nutrition Research Reviews, vol. 23, no. 1, pp. 37–46, 2010.
View at: Publisher Site | Google Scholar
V. Taverniti and S. Guglielmetti, “The immunomodulatory properties of probiotic microorganisms beyond their viability (ghost probiotics: proposal of paraprobiotic concept),” Genes & Nutrition, vol. 6, no. 3, pp. 261–274, 2011.
View at: Publisher Site | Google Scholar
N. Pique, M. Berlanga, and D. Minana-Galbis, “Health benefits of heat-killed (Tyndallized) probiotics: an overview,” International Journal of Molecular Sciences, vol. 20, no. 10, pp. 1–30, 2019.
View at: Google Scholar
D. M. Linares, C. Gómez, E. Renes et al., “Lactic acid bacteria and bifidobacteria with potential to design natural biofunctional health-promoting dairy foods,” vol. 8, no. 846, pp. 1–11, 2017.
View at: Google Scholar
F. Mozzi, “Lactic acid bacteria,” Encyclopedia of Food and Health, vol. 2016, pp. 501–508, 2016.
View at: Publisher Site | Google Scholar
M. A. O. Dawood, S. Koshio, M. Ishikawa, and S. Yokoyama, “Effects of heat killed Lactobacillus plantarum (LP20) supplemental diets on growth performance, stress resistance and immune response of red sea bream, Pagrus major,” Pagrus major, Aquaculture, vol. 442, pp. 29–36, 2015.
View at: Publisher Site | Google Scholar
N. Van Nguyen, S. Onoda, T. Van Khanh et al., “Evaluation of dietary heat-killed Lactobacillus plantarum strain L-137 supplementation on growth performance, immunity and stress resistance of Nile tilapia Oreochromis niloticus,” Aquaculture, vol. 498, pp. 371–379, 2019.
View at: Publisher Site | Google Scholar
T. T. T. Hien, T. T. T. Hoa, P. T. Liem, S. Onoda, and P. M. Duc, “Effects of dietary supplementation of heat-killed Lactobacillus plantarum L-137 on growth performance and immune response of bighead catfish (Clarias macrocephalus),” Aquaculture Reports, vol. 20, p. 100741, 2021.
View at: Publisher Site | Google Scholar
T. T. T. Hien, L. C. T. Tu, H. Saw, S. Carris, N. T. Onoda, and M. P. D. Tuan, “Dietary supplemetation with heat-killed Lactobacillus plantarum L-137 improves growth, immune response, and disease resistance of snakehead (Channa striata), Aquaculture, Aquarium, Conservation & Legislation,” International Journal of the Biofux Society, vol. 14, no. 4, pp. 2229–2240, 2021.
View at: Google Scholar
G. Sagada, N. Gray, L. Wang et al., “Effect of dietary inactivated Lactobacillus plantarum on growth performance, antioxidative capacity, and intestinal integrity of black sea bream Acanthopagrus schlegelii fingerlings.,” Aquaculture, vol. 535, p. 736370, 2021.
View at: Publisher Site | Google Scholar
B. Shan, Y. Liu, N. Song et al., “Genetic diversity and population structure of black sea bream Acanthopagrus schlegelii based on mitochondrial control region sequences: the genetic effect of stock enhancement,” Regional Studies in Marine Science, vol. 35, p. 101188, 2020.
View at: Publisher Site | Google Scholar
E. B. Gonzalez, K. Nagasawa, and T. Umino, “Stock enhancement program for black sea bream Acanthopagrus schlegelii in Hiroshima Bay: monitoring the genetic effects,” Aquaculture, vol. 276, no. 1-4, pp. 36–43, 2008.
View at: Publisher Site | Google Scholar
M. A. Dawood, S. Koshio, M. Ishikawa, and S. Yokoyama, “Interaction effects of dietary supplementation of heat-killed Lactobacillus plantarum and β-glucan on growth performance, digestibility and immune response of juvenile red sea bream, Pagrus major,” Pagrus major, Fish Shellfish Immunol, vol. 45, no. 1, pp. 33–42, 2015.
View at: Publisher Site | Google Scholar
S. Lee, K. Katya, A. Hamidoghli, J. Hong, D. J. Kim, and S. C. Bai, “Synergistic effects of dietary supplementation of Bacillus subtilis WB60 and mannanoligosaccharide (MOS) on growth performance, immunity and disease resistance in Japanese eel, Anguilla japonica,” Anguilla japonica, Fish Shellfish Immunol, vol. 83, pp. 283–291, 2018.
View at: Publisher Site | Google Scholar
H. Van Doan, C. Lumsangkul, S. H. Hoseinifar et al., “Modulation of growth, innate immunity, and disease resistance of Nile tilapia Oreochromis niloticus culture under biofloc system by supplementing pineapple peel powder and Lactobacillus plantarum,” Fish & Shellfish Immunology, vol. 115, pp. 212–220, 2021.
View at: Publisher Site | Google Scholar
S. Murosaki, Y. Yamamoto, K. Ito et al., “Heat-killed Lactobacillus plantarum L-137 suppresses naturally fed antigen- specific IgE production by stimulation of IL-12 production in mice,” Journal of Allergy and Clinical Immunology, vol. 102, no. 1, pp. 57–64, 1998.
View at: Publisher Site | Google Scholar
F. Zhou, J. Shao, R. Xu, J. Ma, and Z. Xu, “Quantitative l-lysine requirement of juvenile black sea bream (Sparus macrocephalus),” Aquaculture Nutrition, vol. 16, no. 2, pp. 194–204, 2010.
View at: Publisher Site | Google Scholar
F. Zhou, J. X. Xiao, Y. Hua, B. O. Ngandzali, and Q. J. Shao, “Dietary l-methionine requirement of juvenile black sea bream (Sparus macrocephalus) at a constant dietary cystine level,” Aquaculture Nutrition, vol. 17, no. 5, pp. 469–481, 2011.
View at: Publisher Site | Google Scholar
AOAC, Official Methods of Analysis of the Association of Analytical Chemistry, Association of Official Analytical Chemists Inc, Washington, DC, 1995.
G. Sagada, J. Chen, B. Shen et al., “Optimizing protein and lipid levels in practical diet for juvenile northern snakehead fish Channa argus,” Animal Nutrition, vol. 3, no. 2, pp. 156–163, 2017.
View at: Publisher Site | Google Scholar
A. C. Elia, M. Prearo, N. Pacini, A. J. Dorr, and M. C. Abete, “Effects of selenium diets on growth, accumulation and antioxidant response in juvenile carp,” Ecotoxicology and Environmental Safety, vol. 74, no. 2, pp. 166–173, 2011.
View at: Publisher Site | Google Scholar
D. Han, S. Xie, M. Liu et al., “The effects of dietary selenium on growth performances, oxidative stress and tissue selenium concentration of Gibel carp Carassius auratus gibelio,” Aquaculture Nutrition, vol. 17, no. 3, pp. e741–e749, 2011.
View at: Publisher Site | Google Scholar
J. Nunez Rodriguez, O. Kah, M. Geffard, and F. Le Menn, “Enzyme-linked immunosorbent assay (ELISA) for sole Solea vulgaris vitellogenin,” Comparative Biochemistry and Physiology, vol. 92, no. 4, pp. 741–746, 1989.
View at: Publisher Site | Google Scholar
P. Y. Li, J. Y. Wang, Z. D. Song et al., “Evaluation of soy protein concentrate as a substitute for fishmeal in diets for juvenile starry flounder Platichthys stellatus,” Aquaculture, vol. 448, pp. 578–585, 2015.
View at: Publisher Site | Google Scholar
K. J. Livak and T. D. Schmittgen, “Analysis of relative gene expression data using real-time quantitative PCR and the 2^{−ΔΔ _C_}_T method,” Methods, vol. 25, no. 4, pp. 402–408, 2001.
View at: Publisher Site | Google Scholar
L. Wang, Y. Sun, B. Xu et al., “Effects of berberine supplementation in high starch diet on growth performance, antioxidative status, immune parameters and ammonia stress response of fingerling black sea bream Acanthopagrus schlegelii,” Aquaculture, vol. 527, p. 735473, 2020.
View at: Publisher Site | Google Scholar
L. Wang, G. Sagada, B. Xu, J. Zhang, Q. Shao, and E. Li, “Influence of dietary berberine on liver immune response and intestinal health of Black Sea bream (Acanthopagrus schlegelii) fed with normal and high-lipid diets,” Aquaculture Nutrition, vol. 2022, pp. 1–15, 2022.
View at: Publisher Site | Google Scholar
A. Wang, C. Ran, Y. Wang et al., “Use of probiotics in aquaculture of China--a review of the past decade,” Fish & Shellfish Immunology, vol. 86, pp. 734–755, 2019.
View at: Publisher Site | Google Scholar
M. J. Zorriehzahra, S. T. Delshad, M. Adel et al., “Probiotics as beneficial microbes in aquaculture: an update on their multiple modes of action: a review,” The Veterinary Quarterly, vol. 36, no. 4, pp. 228–241, 2016.
View at: Publisher Site | Google Scholar
Y. Y. Yan, H. Q. Xia, H. L. Yang, S. H. Hoseinifar, and Y. Z. Sun, “Effects of dietary live or heat-inactivated autochthonousBacillus pumilusSE5 on growth performance, immune responses and immune gene expression in grouperEpinephelus coioides,” Aquaculture Nutrition, vol. 22, no. 3, pp. 698–707, 2016.
View at: Publisher Site | Google Scholar
M. A. O. Dawood, F. I. Magouz, M. F. I. Salem, and H. A. Abdel-Daim, “Modulation of digestive enzyme activity, blood health, oxidative responses and growth-related gene expression in GIFT by heat-killed Lactobacillus plantarum,” Aquaculture, vol. 505, pp. 127–136, 2019.
View at: Publisher Site | Google Scholar
W. N. Xu, D. H. Chen, Q. Q. Chen, and W. B. Liu, “Growth performance, innate immune responses and disease resistance of fingerling blunt snout bream, Megalobrama amblycephala adapted to different berberine-dietary feeding modes,” Megalobrama amblycephala adapted to different berberine-dietary feeding modes, Fish Shellfish Immunol, vol. 68, pp. 458–465, 2017.
View at: Publisher Site | Google Scholar
H. V. Doan, S. H. Hoseinifar, S. Jaturasitha, M. A. O. Dawood, and R. Harikrishnan, “The effects of berberine powder supplementation on growth performance, skin mucus immune response, serum immunity, and disease resistance of Nile tilapia Oreochromis niloticus fingerlings,” Aquaculture, vol. 520, p. 734927, 2020.
View at: Publisher Site | Google Scholar
H. Wang, L. Guan, J. Li, M. Lai, and X. Wen, “The effects of berberine on the gut microbiota in Apc (min/+) mice fed with a high fat diet,” Molecules, vol. 23, no. 9, p. 2298, 2018.
View at: Publisher Site | Google Scholar
X. Zhang, Y. Zhao, M. Zhang et al., “Structural changes of gut microbiota during berberine-mediated prevention of obesity and insulin resistance in high-fat diet-fed rats,” PLoS One, vol. 7, no. 8, article e42529, 2012.
View at: Publisher Site | Google Scholar
M. Sewaka, C. Trullas, A. Chotiko et al., “Efficacy of synbiotic Jerusalem artichoke and Lactobacillus rhamnosus GG- supplemented diets on growth performance, serum biochemical parameters, intestinal morphology, immune parameters and protection against Aeromonas veronii in juvenile red tilapia Oreochromi spp,” Fish & Shellfish Immunology, vol. 86, pp. 260–268, 2019.
View at: Publisher Site | Google Scholar
H. Van Doan, S. Doolgindachbaporn, and A. Suksri, “Effect of Lactobacillus plantarumand Jerusalem artichoke Helianthus tuberosus on growth performance, immunity and disease resistance of Pangasius catfish (Pangasius bocourti,Sauvage 1880),” Aquaculture Nutrition, vol. 22, no. 2, pp. 444–456, 2016.
View at: Publisher Site | Google Scholar
G. Sagada, L. Wang, B. Xu et al., “Effect of dietary inactivatedLactobacillus plantarumand selenomethionine supplementation on growth performance and health-related indices of black sea bream (Acanthopagrus Schlegelii) fingerlings,” Aquaculture Nutrition, vol. 27, no. 5, pp. 1529–1543, 2021.
View at: Publisher Site | Google Scholar
Y. Xia, M. Lu, G. Chen et al., “Effects of dietary Lactobacillus rhamnosus JCM1136 and Lactococcus lactis subsp. lactis JCM5805 on the growth, intestinal microbiota, morphology, immune response and disease resistance of juvenile Nile tilapia, Oreochromis niloticus,” Fish & Shellfish Immunology, vol. 76, pp. 368–379, 2018.
View at: Publisher Site | Google Scholar
C. Fang, M. Ma, H. Ji, T. Ren, and S. D. Mims, “Alterations of digestive enzyme activities, intestinal morphology and microbiota in juvenile paddlefish, Polyodon spathula, fed dietary probiotics,” Polyodon spathula, fed dietary probiotics, Fish Physiol Biochem, vol. 41, no. 1, pp. 91–105, 2015.
View at: Publisher Site | Google Scholar
S. S. Giri, S. S. Sen, and V. Sukumaran, “Effects of dietary supplementation of potential probiotic Pseudomonas aeruginosa VSG-2 on the innate immunity and disease resistance of tropical freshwater fish, Labeo rohita,” Labeo rohita, Fish Shellfish Immunol, vol. 32, no. 6, pp. 1135–1140, 2012.
View at: Publisher Site | Google Scholar
S. S. Giri, V. Sukumaran, and M. Oviya, “Potential probiotic _Lactobacillus plantarum_ VSG3 improves the growth, immunity, and disease resistance of tropical freshwater fish, Labeo rohita,” Fish Shellfish Immunology, vol. 34, no. 2, pp. 660–666, 2013.
View at: Publisher Site | Google Scholar
D. R. Tocher, G. Mourente, A. Van Der Eecken et al., “Effects of dietary vitamin E on antioxidant defence mechanisms of juvenile turbot Scophthalmus maximusL halibut Hippoglossus hippoglossusL and sea bream Sparus aurataL,” Aquaculture Nutrition, vol. 8, no. 3, pp. 195–207, 2002.
View at: Publisher Site | Google Scholar
L. Qin, J. Xiang, F. Xiong et al., “Effects of Bacillus licheniformis on the growth, antioxidant capacity, intestinal barrier and disease resistance of grass carp Ctenopharyngodon idella,” Fish & Shellfish Immunology, vol. 97, pp. 344–350, 2020.
View at: Publisher Site | Google Scholar
L. Weifen, Z. Xiaoping, S. Wenhui et al., “Effects of bacillus preparations on immunity and antioxidant activities in grass carp (Ctenopharyngodon idellus),” Fish Physiology and Biochemistry, vol. 38, no. 6, pp. 1585–1592, 2012.
View at: Publisher Site | Google Scholar
C. N. Zhang, X. F. Li, W. N. Xu et al., “Combined effects of dietary fructooligosaccharide and Bacillus licheniformis on innate immunity, antioxidant capability and disease resistance of triangular bream Megalobrama terminalis,” Fish & Shellfish Immunology, vol. 35, no. 5, pp. 1380–1386, 2013.
View at: Publisher Site | Google Scholar
K. L. Lu, L. N. Wang, D. D. Zhang, W. B. Liu, and W. N. Xu, “Berberine attenuates oxidative stress and hepatocytes apoptosis via protecting mitochondria in blunt snout bream Megalobrama amblycephala fed high-fat diets,” Fish Physiology and Biochemistry, vol. 43, no. 1, pp. 65–76, 2017.
View at: Publisher Site | Google Scholar
J. M. Hur, M. S. Hyun, S. Y. Lim, W. Y. Lee, and D. Kim, “The combination of berberine and irradiation enhances anti-cancer effects via activation of p38 MAPK pathway and ROS generation in human hepatoma cells,” Journal of Cellular Biochemistry, vol. 107, no. 5, pp. 955–964, 2009.
View at: Publisher Site | Google Scholar
Y. Wang, Y. Wu, Y. Wang et al., “Antioxidant properties of probiotic bacteria,” Nutrients, vol. 9, no. 5, p. 521, 2017.
View at: Publisher Site | Google Scholar
L. X. Wang, K. Liu, D. W. Gao, and J. K. Hao, “Protective effects of two Lactobacillus plantarum strains in hyperlipidemic mice,” World Journal of Gastroenterology, vol. 19, no. 20, pp. 3150–3156, 2013.
View at: Publisher Site | Google Scholar
P. Dinesh and M. Rasool, “Berberine, an isoquinoline alkaloid suppresses TXNIP mediated NLRP3 inflammasome activation in MSU crystal stimulated RAW 264.7 macrophages through the upregulation of Nrf2 transcription factor and alleviates MSU crystal induced inflammation in rats,” International Immunopharmacology, vol. 44, pp. 26–37, 2017.
View at: Publisher Site | Google Scholar
A. K. Ray, K. Ghosh, and E. Ringø, “Enzyme-producing bacteria isolated from fish gut: a review,” Aquaculture Nutrition, vol. 18, no. 5, pp. 465–492, 2012.
View at: Publisher Site | Google Scholar
H. Liu, S. Wang, Y. Cai et al., “Dietary administration of _Bacillus subtilis HAINUP40 enhances growth, digestive enzyme activities, innate immune responses and disease resistance of tilapia, Oreochromis niloticus,” Fish & shellfish Immunology, vol. 60, pp. 326–333, 2017.
View at: Publisher Site | Google Scholar
S. Won, A. Hamidoghli, W. Choi et al., “Effects of Bacillus subtilis WB60 and Lactococcus lactis on growth, immune responses, histology and gene expression in Nile tilapia, Oreochromis niloticus,” vol. 8, no. 1, pp. 67–82, 2020.
View at: Publisher Site | Google Scholar
S. K. Allameh, E. Ringø, F. M. Yusoff, H. M. Daud, and A. Ideris, “Dietary supplement ofEnterococcus faecalison digestive enzyme activities, short-chain fatty acid production, immune system response and disease resistance of Javanese carp (Puntius gonionotus,Bleeker 1850),” Aquaculture Nutrition, vol. 23, no. 2, pp. 331–338, 2017.
View at: Publisher Site | Google Scholar
M. C. Holland and J. D. Lambris, “The complement system in teleosts,” Fish & Shellfish Immunology, vol. 12, no. 5, pp. 399–420, 2002.
View at: Publisher Site | Google Scholar
Y. Z. Sun, H. L. Yang, R. L. Ma, and W. Y. Lin, “Probiotic applications of two dominant gut Bacillus strains with antagonistic activity improved the growth performance and immune responses of grouper Epinephelus coioides,” Fish & Shellfish Immunology, vol. 29, no. 5, pp. 803–809, 2010.
View at: Publisher Site | Google Scholar
H. Li, C. Fan, H. Lu et al., “Protective role of berberine on ulcerative colitis through modulating enteric glial cells-intestinal epithelial cells-immune cells interactions,” Acta Pharmaceutica Sinica B, vol. 10, no. 3, pp. 447–461, 2020.
View at: Publisher Site | Google Scholar
J. Zhao, Y. Wang, X. Wu et al., “Inhibition of CCL19 benefits non‑alcoholic fatty liver disease by inhibiting TLR4/NF‑κB‑p65 signaling,” Molecular Medicine Reports, vol. 18, no. 5, pp. 4635–4642, 2018.
View at: Publisher Site | Google Scholar
T. Alenghat, L. C. Osborne, S. A. Saenz et al., “Histone deacetylase 3 coordinates commensal-bacteria-dependent intestinal homeostasis,” Nature, vol. 504, no. 7478, pp. 153–157, 2013.
View at: Publisher Site | Google Scholar
J. Mao, S. Qi, Y. Cui et al., “Lactobacillus rhamnosus GG attenuates lipopolysaccharide-induced inflammation and barrier dysfunction by regulating MAPK/NF-κB signaling and modulating metabolome in the piglet intestine,” The Journal of Nutrition, vol. 150, no. 5, pp. 1313–1323, 2020.
View at: Publisher Site | Google Scholar
K. W. Moore, R. de Waal Malefyt, R. L. Coffman, and A. O'Garra, “Interleukin-10 and the interleukin-10 receptor,” Annual Review of Immunology, vol. 19, no. 1, pp. 683–765, 2001.
View at: Publisher Site | Google Scholar
S. S. Iyer and G. Cheng, “Role of interleukin 10 transcriptional regulation in inflammation and autoimmune disease,” Critical Review in Immunology, vol. 32, no. 1, pp. 23–63, 2012.
View at: Publisher Site | Google Scholar
W. J. Jang, J. M. Lee, M. T. Hasan, B. J. Lee, S. G. Lim, and I. S. Kong, “Effects of probiotic supplementation of a plant-based protein diet on intestinal microbial diversity, digestive enzyme activity, intestinal structure, and immunity in olive flounder Paralichthys olivaceus,” Fish & Shellfish Immunology, vol. 92, pp. 719–727, 2019.
View at: Publisher Site | Google Scholar
A. M. Hamdan, A. F. El-Sayed, and M. M. Mahmoud, “Effects of a novel marine probiotic, lactobacillus plantarum AH 78, on growth performance and immune response of Nile tilapia Oreochromis niloticus,” Journal of Applied Microbiology, vol. 120, no. 4, pp. 1061–1073, 2016.
View at: Publisher Site | Google Scholar
C. Chen, Z. Yu, Y. Li, J. Fichna, and M. Storr, “Effects of berberine in the gastrointestinal tract - a review of actions and therapeutic implications,” The American Journal of Chinese Medicine, vol. 42, no. 5, pp. 1053–1070, 2014.
View at: Publisher Site | Google Scholar
B.-H. Choi, I.-S. Ahn, Y.-H. Kim et al., “Berberine reduces the expression of adipogenic enzymes and inflammatory molecules of 3T3-L1 adipocyte,” Experimental and Molecular Medicine, vol. 38, no. 6, pp. 599–605, 2006.
View at: Publisher Site | Google Scholar
T. Lou, Z. Zhang, Z. Xi et al., “Berberine inhibits inflammatory response and ameliorates insulin resistance in hepatocytes,” Inflammation, vol. 34, no. 6, pp. 659–667, 2011.
View at: Publisher Site | Google Scholar

Copyright

Copyright © 2022 Gladstone Sagada et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

PDF Download Citation

Download other formats

Order printed copies

Views

483

Downloads

466

Citations