The synergistic effects of plant polysaccharide and Pediococcus acidilactici as a synbiotic additive on growth, antioxidant status, immune response, and resistance of Nile tilapia (Oreochromis niloticus) against Aeromonas hydrophila.

This study evaluated the growth performance, immune responses, and disease resistance of Nile tilapia upon pistachio hulls derived polysaccharide (PHDP) and Pediococcus acidilactici (PA) separately or as synbiotic. Fish received four types of diets: T1, control; T2, PHDP (0.1%); T3, PA (0.2%); T4, PHDP (0.1%) +PA (0.2%) for 56 days. The results showed that final weight and weight gain were markedly higher in fish fed T4 diet than that given T1 and T2 diets (P ≤ 0.05). In addition, a significantly greater specific growth rate was obtained by the T4 diet compared to the control. Fish survival was significantly improved in all supplemented diets compared to the control. On the other hand, the activities of lipase, protease, and amylase showed significant increases in the T4 group compared with other feeding groups. The total leucocytes and lymphocytes proportion significantly elevated in T3 and T4 than remaining groups (P ≤ 0.05). Further, fish fed T3 diet presented significantly higher serum total protein, total immunoglobulin, lysozyme activity (LYZ), alternative complement activity (ACH50), and alkaline phosphatase activity compared to fish fed T1 and T2 diets, while the mentioned indices were found significantly highest in T4 group than others. Fish received T3 and T4 diets had higher skin mucus LYZ and ACH50 than those fed T1 and T2 diets (P ≤ 0.05). The malondialdehyde levels were significantly declined in T3 and T4 when compared to the control. Fish fed T3 and T4 diets demonstrated significantly enhanced superoxide dismutase, catalase, and glutathione peroxidase activities compared to the control. The intestinal propionic acid significantly increased by T2 and T4 diets, while the highest levels of acetic acid detected in fish given T4 diet. The expression levels of tumor necrosis factor α (TNF-α), interleukin 1ß (IL-1ß), and interleukin 10 (IL-10) were significantly affected by T3 and T4 supplements. The efficacy of T4 diet against Aeromonas hydrophila infection was documented by a significantly lower mortality rate. In conclusion, the combination of PHDP and PA presented promising results as a synbiotic feed additive for Nile tilapia.

Pistachio hull polysaccharide protects Nile tilapia against LPS-induced excessive inflammatory responses and oxidative stress, possibly via TLR2 and Nrf2 signaling pathways.

Polysaccharides are polymeric carbohydrates found in living organisms, which have several physiological functions. In the present study, Nile tilapia (Oreochromis niloticus) were fed diets containing three levels (0%, 0.2%, and 0.6%) of Pistacia vera hull polysaccharide (PHP) for 45 days and then injected with PBS or bacterial lipopolysaccharide (LPS). Before the LPS challenge, Nile tilapia fed 0.2% and 0.6% PHP showed significantly increased mean final weight and weight gain compared to those received 0% PHP. The specific growth rate and feed conversion ratio were significantly improved in the treatment fed 0.6% PHP compared to the remaining groups. After LPS challenge, the activities of liver antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase exhibited the highest values in the 0.6% PHP group. Malondialdehyde (MDA) levels were significantly augmented in the model (fed 0% PHP diet and injected with LPS) and 0.2% PHP groups compared to the control. However, MDA showed decreased levels in the 0.6% PHP group. LPS induced higher mRNA and/or protein levels of Toll-like receptor 2 (TLR2), nuclear factor kappa B (NF-κB), myeloid differentiation primary response protein 88 (Myd88), tumor necrosis factor α (TNF-α), interleukin 1ß (IL-1ß), and interferon γ (IFN-γ) in Nile tilapia liver. However, PHP administration significantly upregulated the expression of interleukin 10 (IL-10), nuclear erythroid 2-related factor 2 (Nrf2), SOD, and CAT, but markedly suppressed TLR2, NF-κB, Myd88, and pro-inflammatory cytokine expression and/or production in the liver. The findings of the current study indicated that PHP has positive effects on growth performance, immune gene-related expression, and antioxidative activities. We can conclude that PHP can attenuate LPS-induced oxidative stress and inflammatory responses in vivo, possibly via induction of Nrf2 and blockade of TLR2/Myd88/NF-κB pathways in Nile tilapia.

The growth performance, antioxidant capacity, immunological responses, and the resistance against Aeromonas hydrophila in Nile tilapia (Oreochromis niloticus) fed Pistacia vera hulls derived polysaccharide.

The present study aimed at evaluating the supplementation of dietary Pistacia vera hulls derived polysaccharide (PHDP) at varying levels (0, 2.5, 5, and 10 g/kg diet) on the growth rate, digestive enzyme activity, immune response, and antioxidative capacity of Nile tilapia. After the feeding trial (60 days), fish were injected intraperitoneally with Aeromonas hydrophila, and the cumulative mortality was calculated for 10 days. The final body weight, weight gain, specific growth rate, survival rate, amylase activity, red blood cells, hemoglobin, serum total protein, and serum lysozyme activity (LZM) were significantly higher in fish fed PHDP at 5 and 10 g/kg than fish fed 0 and 2.5 g/kg diet (P ≤ 0.05). However, the feed conversion ratio and glucose levels were significantly decreased by 5 and 10 g PHDP/kg diet. Further, the protease digestive activity, serum alkaline phosphatase, and serum alternative complement (ACH50) had higher values in fish fed 5 g PHDP/kg diet than tilapia fed 0 g PHDP/kg diet (P ≤ 0.05). The serum protease and immunoglobulin (Ig) presented the highest values in fish fed 10 g PHDP/kg diet followed by fish fed 5 g PHDP/kg diet (P ≤ 0.05). The LZM, ACH50, and Ig in the mucus samples were significantly higher in fish fed 5 or 10 g PHDP/kg diet than fish fed 0 g PHDP/kg diet (P ≤ 0.05). The levels of superoxide dismutase and catalase in the liver tissue was significantly higher in fish fed 5 or 10 g PHDP/kg diet than fish fed 2.5 g PHDP/kg diet (P ≤ 0.05). On the other hand, malondialdehyde concentration was significantly lower in tilapia fed 5 and 10 g PHDP/kg diet than tilapia fed 0 and 2.5 g PHDP/kg diet (P ≤ 0.05). The lowest mortality rate and the highest relative percentage survival were in fish fed 5 g/kg followed by 10 g level after 10 days of A. hydrophila injection. Accordingly, dietary PHDP at 5-10 g/kg is recommended to improve the growth performance, antioxidative capacity, immune response, and resistance against A. hydrophila in Nile tilapia.

Effects of dietary Lactobacillus plantarum (KC426951) in biofloc and stagnant-renewal culture systems on growth performance, mucosal parameters, and serum innate responses of Nile tilapia Oreochromis niloticus.

Effects of dietary Lactobacillus plantarum (KC426951) on growth and innate responses of Nile tilapia Oreochromis niloticus were evaluated in biofloc technology system and stagnant-renewal culture system (SRCS). The 90-day-long experiment contained four treatments: SRCS without probiotic (T1), SRCS with probiotic (T2), biofloc without probiotic (T3), and biofloc with probiotic (T4). The administration dose of probiotic was 2 × 108 CFU kg-1 diet. At the end of experiment, the mean final weights, specific growth rates, feed conversion ratios, and total biomass were significantly (P < 0.05) better in BFT treatments, with no significant effect of probiotic on these parameters in both culture systems. Meanwhile, skin mucosal parameters including total protein (TP), lysozyme (LYZ), alkaline phosphatase (ALP), and protease (PRO) activity were significantly enhanced following probiotic supplementation. T4 treatment displayed a significantly higher LYZ and ALP activity in mucus versus other treatments. Also, serum alternative complement activity was significantly heightened in probiotic-supplemented fish. Superoxide dismutase activity in T4 was detected higher than that of SRCS groups. The results of the current study demonstrated the enhancement of some mucosal and serum innate responses of Nile tilapia in both culture systems upon L. plantarum (KC426951) supplementation.

The effects of silver nanoparticles (Ag-NPs) sublethal concentrations on common carp (Cyprinus carpio): Bioaccumulation, hematology, serum biochemistry and immunology, antioxidant enzymes, and skin mucosal responses.

The present study aimed to evaluate the effects of different waterborne sublethal concentrations of Ag-NPs LC50 (96h) on common carp Cyprinus carpio using a multi-biomarker approach. Fish (9.22 ± 0.12 g) were stocked in fiberglass tanks and exposed to concentrations of 0 (control), 12.5%, 25% and 50% of Ag-NPs LC50 (96h) or Ag-NO3 LC50 (96h), as the source of Ag+ ion, for a period of 21 days. At the end of study, tissue Ag contents were significantly (P < 0.05) higher and different in fish exposed to concentrations of 25% and 50% compared to the control. The numbers of RBCs, hematocrit, and MCHC values at these concentrations differed significantly in respect to the control. No significant effects were observed for hemoglobin, MCH, and MCV values. The number of WBCs was significantly higher at concentrations of 12.5% and 25% compared to the control. Meanwhile, the percentage of neutrophils significantly elevated at concentrations of 25% and 50%. Serum total protein at concentration of 50% detected significantly lower than that of 12.5% or the control. The serum albumin and globulin levels significantly declined in Ag-NPs-exposed groups versus the control. The serum ACH50 and total immunoglobulins showed significantly lower values in the treatments of 25% and 50% compared to the control. The serum glucose, cortisol, ALT, and ALP values significantly escalated upon Ag-NPs exposure. The serum SOD and CAT showed enhanced activity in the treatment of 12.5% vice versa significantly diminished at concentrations of 25% and 50% compared to the control. The exposure to the concentrations of 25% and 50% significantly dwindled the lysozyme activity and total immunoglobulin levels in skin mucus. In conclusion, sublethal concentrations of Ag-NPs LC50 (96h) impaired fish health status at higher concentrations and 12.5% of Ag-NPs LC50 (96h) was presumably safe for common carp aquaculture.

Ginger (Zingiber officinale) extract affects growth performance, body composition, haematology, serum and mucosal immune parameters in common carp (Cyprinus carpio).

The present study was designed to investigate the dietary effects of ginger extract (Zingiber officinale) on common carp (Cyprinus carpio). Three hundred and sixty fish weighing 10.9 ± 0.17 g were randomly divided into four experimental treatments in triplicates. Four experimental diets were designed containing different inclusion levels of ginger extract as 0% (control), 0.1%, 0.2%, and 0.4% named T1, T2, T3, and T4, respectively, and fish were fed 3% of body weight daily for 60 days. Feed conversion ratio, weight gain, and specific growth rate parameters were measured higher in the groups fed with ginger extract compared to the control, especially in T3 (P < 0.05). Also, fish treated with ginger extract showed lower carcass moisture and lipid, but higher protein contents compared to the control. The results showed significant increase in the number of erythrocytes and leucocytes, as well as the level of hematocrit and hemoglobin in fish fed with supplemented diets. In addition, significant enhancement was observed in cases of serum parameters, including total protein, albumin, globulin, lysozyme, superoxide dismutase, catalase, and total immunoglobulins in T3 and T4 compared to the control. The lowest serum cortisol level was noticed in T3. Furthermore, the T3 group showed the highest activities of skin mucus lysozyme, alkaline phosphatase, and protease (P < 0.05). In conclusion, the present results demonstrated that Zingiber officinale extract at a 0.2% level can effectively improve the growth and health status of Common Carp (Cyprinus carpio).

Effect of different types and levels of fat addition and pellet binders on physical pellet quality of broiler feeds.

Two experiments were conducted to evaluate the effects of different types and levels of mixer-added fat (soybean oil: SO and calcium fat powder: CFP) and pellet binders (PBs: calcium lignosulfonate (CaLS) and bentonite (Ben)) on physical pellet quality (PPQ) parameters. PPQ included pellet durability index (PDI), pellet hardness, and pellet length of broiler diets processed under short-term conditioning. The first experiment had 4 treatments arranged as a 2 × 2 factorial with 2 types (SO and CFP) and 2 levels (1.5 and 3%) of mixer-added fat. In the second experiment, 22 treatments, combinations of 2 types of mixer-added fat (SO and CFP) at 3 levels (0, 1.5 and 3%) and 2 types of PB (CaLS = 0, 0.5, and 1% and Ben = 0, 1, and 2%), were arranged by a completely randomized design. PDI was measured by 2 devices: Pfost Tumbling box (PDIT) and Holmen NHP tester (PDIH). The results showed that the diets containing 1.5% CFP without PB had significant differences in all PPQ parameters. The results revealed that adding 0.5% CaLS to the 3% SO diets significantly enhanced PDIH, pellet hardness, and pellet length compared to other treatments. Moreover, 1.5% CFP diets with 2% Ben had significantly higher PDIT, PDIH, and pellet hardness among the treatments. Based on contour plots, different levels of Ben in the diets containing SO failed to create optimum PDIT values (>96%). However, 1.5 to 2.50% CFP diets without Ben had the optimum PDIT values. The optimum PDIT value was achieved by the diets containing 3% SO in the range of 0.21 to 0.56% CaLS. Furthermore, adding 0.5% CaLS to the diets containing less than 2.86% SO resulted in suboptimal PDIT values (<96%). The diets containing 1.5 to 2.50% CFP without CaLS had the optimum PDIT values. However, increasing CaLS levels more than 0.38% led to suboptimal PDIT values. Overall, these results indicated that the selection of appropriate PBs should be based on type and level of mixer-added fat.

Effects of feed form and particle size, and pellet binder on performance, digestive tract parameters, intestinal morphology, and cecal microflora populations in broilers.

This study was conducted to assess the effects of feed form (FF), particle size (PS), and pellet binder (PB) on performance, digestive tract parameters, intestinal morphology, and cecal microflora populations in broiler chicks. Two FF (mash and pellet), two feed PS (fine and coarse), and two levels of PB (0 and 3%) were evaluated in a completely randomized design with a 2 × 2 × 2 factorial arrangement of treatments. A total of 256 one-day-old male Ross 308 chicks were allocated into 8 treatments consisting of 8 replicates (4 birds/cage) with equal weight from 25 to 42 d of age. The results showed a three-way interaction among FF, PS, and PB for growth performance in broilers. The growth performance parameters were significantly improved by feeding the pellet-coarse PS-3% PB diet in comparison to the other treatments (P < 0.05). Birds fed pelleted diets had lower relative weight of the gizzard and pancreas, goblet cell (GC) number of the duodenum, crypt depth (CD) of the jejunum, higher villus height (VH) of the duodenum, and cecal spore-forming bacteria population (P < 0.05). Coarse PS resulted in lower jejunum CD and higher cecal count of lactic acid bacteria (LAB) and spore-forming bacteria (P < 0.05). Adding PB significantly improved pellet durability index (PDI) (P < 0.05). In addition, coarse PS-3% PB diets had higher pellet hardness. Also, adding PB to diets significantly (P < 0.05) decreased cecal spore-forming bacteria population. Pellet-coarse PS-3% PB diets fed broilers had a lower (P < 0.05) cecal Clostridium spp. bacterial population. The main factors had no significant effect (P > 0.05) on cecal Coliform spp. levels. Birds fed pellet-3% PB diets had higher VH:CD ratio both in the duodenum and jejunum, and those fed mash-3% PB diets had significantly (P < 0.05) lower VH in the jejunum than other treatments. Overall, pellet-coarse PS-3% PB diets improved growth performance through the positive changes on intestinal morphology of broilers under short-term conditioning (STC).

Metabolism of [U-(13)C]aspartate by astroglial cultures: nuclear magnetic resonance analysis of the culture media.

In brain the amino acid L-aspartate serves roles as: (1) putative transmitter, (2) protein precursor, (3) donor of atoms for the biosynthesis of pyrimidine and purine bases, and (4) fuel for energy metabolism. Astrocytes dominate aspartate clearance in brain, and in culture they take up aspartate and quickly metabolize it. In brain, only astrocytes were shown to express the enzymes for de novo pyrimidine biosynthesis. To gain more details about the spectrum of metabolites generated from aspartate and subsequently released by cultured astrocytes a (13)C-nuclear magnetic resonance analysis was performed of [U-(13)C]aspartate supplemented incubation media exposed to astroglial cultures. The results show that astrocytes readily metabolize aspartate and release into their culture media (13)C-isotopomers of lactate, glutamine, citrate and alanine. Despite the presence in astroglial cells of two tandem enzymes of pyrimidine biosynthesis and their mRNAs, pyrimidine nucleotide-related heterocyclic compounds such as dihydroorotate and orotate could not be detected in the culture media.

Glial metabolism of valine.

The three essential amino acids, valine, leucine and isoleucine, constitute the group of branched-chain amino acids (BCAAs). BCAAs are rapidly taken up into the brain parenchyma, where they serve several distinct functions including that as fuel material in brain energy metabolism. As one function of astrocytes is considered the production of fuel molecules that support the energy metabolism of adjacent neural cells in brain. Astroglia-rich primary cultures (APC) were shown to rapidly dispose of the BCAAs, including valine, contained in the culture medium. While the metabolisms of leucine and isoleucine by APC have already been studied in detail, some aspects of valine metabolism remained to be determined. Therefore, in the present study an NMR analysis was performed to identify the (13)C-labelled metabolites that are generated by APC during catabolism of [U-(13)C]valine and that are subsequently released into the incubation medium. The results presented show that APC (1) are potently disposing of the valine contained in the incubation medium; (2) are capable of degrading valine to the tricarboxylic acid (TCA) cycle member succinyl-CoA; and (3) release into the extracellular milieu valine catabolites and compounds generated from them such as [U-(13)C]2-oxoisovalerate, [U-(13)C]3-hydroxyisobutyrate, [U-(13)C]2-methylmalonate, [U-(13)C]isobutyrate, and [U-(13)C]propionate as well as several TCA cycle-dependent metabolites including lactate.

Glial metabolism of isoleucine.

Isoleucine, together with leucine and valine, constitutes the group of branched-chain amino acids (BCAAs). BCAAs are transported from the blood into the brain parenchyma, where they can serve several distinct functions. Since brain tissue is known to oxidatively metabolize BCAAs to CO(2), they are considered as fuel material in brain energy metabolism. Also, in the case of leucine, cultured astrocytes have been reported to be able to completely oxidize BCAA. While the metabolism of leucine by astroglia-rich primary culture (APC) has already been studied in detail, the metabolic fates of isoleucine and valine in these cells remained to be identified. Therefore, in the present study an NMR analysis was performed of (13)C-labelled metabolites generated in the catabolism of [U-(13)C]Ile by astrocytes and released by them into the incubation medium. APC potently removed isoleucine from the medium and metabolized it. The major isoleucine metabolites released from APC are 2-oxo-3-methylvalerate, 2-methylbutyrate, 3-hydroxy-2-methylbutyrate and propionate. To a lesser extent, APC generate and release also [2,3-(13)C]glutamine, [4,5-(13)C]glutamine and (13)C-labelled isotopomers of lactate and citrate. These results show that APC can release into the extracellular milieu catabolites and several TCA cycle dependent metabolites resulting from the degradation of isoleucine.