Nigella sativa Extract Potentially Inhibited Methicillin Resistant Staphylococcus aureus Induced Infection in Rabbits: Potential Immunomodulatory and Growth Promoting Properties.

Weaning is the most crucial period associated with increased stress and susceptibility to diseases in rabbits. Methicillin-resistant Staphylococcus aureus (MRSA), a historic emergent pathogen related to post weaning stressors, adversely affects rabbit's growth rate and productive cycle. Since MRSA is rapidly evolving antibiotics resistance, natural products are desperately required to tackle the public health threats posed by antimicrobial resistance. Thus, this study aimed to screen the iin vitro antibacterial activity of Nigella sativa extract (NSE) and its interactions with antibiotics against MRSA isolates. Moreover, 200 weaned rabbits were divided into 4 groups to investigate the iin vivo superiority of NSE graded levels towards growth performance, tight junction integrity, immune responsiveness and resistance against MRSA. Herein, NSE showed promising antimicrobial activities against MRSA isolates from animal (77.8%) and human (64.3%) origins. Additionally, MRSA isolates exposed to NSE became sensitive to all antimicrobials to which they were previously resistant. Our results described that the growth-promoting functions of NSE, especially at higher levels, were supported by elevated activities of digestive linked enzymes. Post-NSE feeding, rabbits' sera mediated bactericidal activities against MRSA. Notably, upregulated expression of occludin, CLDN-1, MUC-2 and JAM-2 genes was noted post NSE supplementation with maximum transcriptional levels in 500 mg/kg NSE fed group. Our data described that NSE constitutively motivated rabbits' immune responses and protected them against MRSA-induced experimental infection. Our results suggest the antimicrobial, growth stimulating and immunomodulation activities of NSE to maximize the capability of rabbits for disease response.

Modulatory Impacts of Multi-Strain Probiotics on Rabbits' Growth, Nutrient Transporters, Tight Junctions and Immune System to Fight against Listeria monocytogenes Infection.

Multi-strain probiotics (MSP) are considered innovative antibiotics' substitutes supporting superior gut health and immunity of farmed rabbits. The promising roles of MSP on performance, intestinal immunity, integrity and transporters, and resistance against Listeria monocytogenes (L. monocytogenes) were evaluated. In the feeding trial, 220 rabbits were fed a control diet or diet supplemented with three MSP graded levels. At 60 days of age, rabbits were experimentally infected with L. monocytogenes and the positive control, enrofloxacin, prophylactic MSP (MSPP), and prophylactic and therapeutic MSP (MSPTT) groups were included. During the growing period, MSP at the level of 1 × 108 CFU/kg diet (MSPIII) promoted the rabbits' growth, upregulated the nutrient transporters and tight-junction-related genes, and modified cytokines expression. Supplementing MSPTT for L. monocytogenes experimentally-infected rabbits restored the impaired growth and intestinal barriers, reduced clinical signs of severity and mortalities, and attenuated the excessive inflammatory reactions. Notably, enrofloxacin decreased L. monocytogenes and beneficial microbial loads; unlike MSPTT, which decreased pathogenic bacterial loads and sustained the beneficial ones. Histopathological changes were greatly reduced in MSPTT, confirming its promising role in restricting L. monocytogenes translocation to different organs. Therefore, our results suggest the use of MSPTT as an alternative to antibiotics, thereby conferring protection for rabbits against L. monocytogenes infection.

Therapeutic Potential of Quercetin Loaded Nanoparticles: Novel Insights in Alleviating Colitis in an Experimental DSS Induced Colitis Model.

Oxidative stress is considered the main etiologic factor involved in inflammatory bowel disease (IBD). Integration of nanocarriers for natural therapeutic agents with antioxidant and anti-inflammatory potential is a novel promising candidate for curing IBD. Herein, the colonic antioxidant and anti-inflammatory effects of different concentrations of quercetin nanoparticles (QT-NPs) were evaluated using a dextran sulfate sodium (DSS)-induced colitis model. Following colitis induction, the efficacy and mechanistic actions of QT-NPs were evaluated by assessing lesion severity, molecular aids controlling oxidative stress and inflammatory response, and histopathological and immunohistochemistry examination of colonic tissues. Administration of QT-NPs, especially at higher concentrations, significantly reduced the disease activity index and values of fecal calprotectin marker compared to the colitic group. Colonic oxidant/antioxidant status (ROS, H2O2, MDA, SOD, CAT, GPX and TAC) was restored after treatment with higher concentrations of QT-NPs. Moreover, QT-NPs at levels of 20 mg/kg and, to a lesser extent, 15 mg/kg reduced Nrf2 and HO-1 gene expression, which was in line with decreasing the expression of iNOS and COX2 in colonic tissues. Higher concentrations of QT-NPs greatly downregulated pro-inflammatory cytokines; upregulated genes encoding occludin, MUC-2 and JAM; and restored the healthy architectures of colonic tissues. Taken together, these data suggest that QT-NPs could be a promising alternative to current IBD treatments.

Dietary Eugenol Nanoemulsion Potentiated Performance of Broiler Chickens: Orchestration of Digestive Enzymes, Intestinal Barrier Functions and Cytokines Related Gene Expression With a Consequence of Attenuating the Severity of E. coli O78 Infection.

Recently, the use of essential oils (EOs) or their bioactive compounds encapsulated by nanoparticles as alternative supplements for in-feed antimicrobials is gaining attention, especially in organic poultry production. Focusing on eugenol, its incorporation into the nanoformulation is a novel strategy to improve its stability and bioavailability and thus augment its growth-boosting and antimicrobial activities. Therefore, we explored eugenol nanoemulsion activities in modulating growth, digestive and gut barrier functions, immunity, cecal microbiota, and broilers response to avian pathogenic E. coli challenge (APEC) O78. A total of 1,000 one-day-old broiler chicks were allocated into five groups; negative control (NC, fed basal diet), positive control (PC), and 100, 250, and 400 mg/kg eugenol nanoemulsion supplemented groups. All groups except NC were challenged with APEC O78 at 14 days of age. The results showed that birds fed eugenol nanoemulsion displayed higher BWG, FI, and survivability and most improved FCR over the whole rearing period. Birds fed 400 mg/kg of eugenol nanoemulsion sustained a higher growth rate (24% vs. PC) after infection. Likely, the expression of digestive enzymes' genes (AMY2A, CCK, CELA1, and PNLIP) was more prominently upregulated and unaffected by APEC O78 challenge in the group fed eugenol nanoemulsion at the level of 400 mg/kg. Enhanced gut barrier integrity was sustained post-challenge in the group supplemented with higher levels of eugenol nanoemulsion as evidenced by the overexpression of cathelicidins-2, ß-defensin-1, MUC-2, JAM-2, occludin, CLDN-1, and FABP-2 genes. A distinct modulatory effect of dietary eugenol nanoemulsion was observed on cytokine genes (IL-1ß, TNF-α, IL-6, IL-8, and IL-10) expression with a prominent reduction in the excessive inflammatory reactions post-challenge. Supplementing eugenol nanoemulsion increased the relative cecal abundance of Lactobacillus species and reduced Enterobacteriaceae and Bacteriods counts. Notably, a prominent reduction in APEC O78 loads with downregulation of papC, iroN, iutA, and iss virulence genes and detrimental modifications in E. coli morphological features were noticed in the 400 mg/kg eugenol nanoemulsion group at the 3rd-week post-challenge. Collectively, we recommend the use of eugenol nanoemulsion as a prospective targeted delivery approach for achieving maximum broilers growth and protection against APEC O78 infection.

Immunosuppressive Effects of Thallium Toxicity in Nile Tilapia Fingerlings: Elucidating the Rescue Role of Astragalus membranaceus Polysaccharides.

This study evaluated the immunotoxic effects of thallium (Tl) in Nile tilapia fingerlings and the recovery role of dietary Astragalus membranaceus polysaccharides (ASs). An 8-week experiment was designed where 180 fishes were randomly and equally assigned in triplicates into the six groups: the control group (CNT) was reared in unpolluted water and fed a commercial diet, two groups were fed a well-balanced commercial diet plus 1.5 and 3.0 g AS/kg diet (AS0.15 and AS0.30), respectively, the fourth group was exposed to a sublethal dose of Tl (41.9 µg l-1) [equal to 1/10 of 96-h lethal concentration 50 (LC50)], and the last two groups were fed 0.15 and 0.3% AS, respectively, and concurrently exposed to Tl (41.9 µg l-1) (AS0.15+Tl and AS0.30+Tl). Fish hematobiochemical parameters, serum immunity [nitric oxide, total immunoglobulin M (IgM) levels, and lysozyme activity], transcription of hepatic interferon-γ (IFN-γ), interleukin-1ß (IL-1ß), and tumor necrosis factor-α (TNF-α), and resistance to Aeromonas hydrophila (A. hydrophila) were assessed. Hematobiochemical parameters and serum immune indices were significantly decreased in the fish group exposed to sublethal Tl concentration compared to the CNT group. Furthermore, Tl exposure significantly induced overexpression of IL-1ß, TNF-α, and IFN-γ genes (4.22-, 5.45-, and 4.57-fold higher, respectively) compared to CNT values. Tl exposure also increased the cumulative mortality (%) in Nile tilapia challenged with A. hydrophila. Remarkably, the groups fed AS0.15+Tl and AS0.30+Tl significantly ameliorated all the aforementioned parameters, but did not reach CNT values. Our findings suggest the possible immunomodulating roles of dietary AS in recovering the immunotoxic effects of Tl in Nile tilapia. We can conclude that dietary AS would be useful for maintaining the immunity of Nile tilapia fingerlings.

Early nutrition programming with Astragalus membranaceus polysaccharide: its effect on growth, carcasses, immunity, antioxidants, lipid profile and liver and kidney functions in broiler chickens.

This study aimed to determine the influence of in ovo injection of Astragalus membranaceus polysaccharide on growth, feed consumption, feed conversion ratio, carcasses, hematology, and blood metabolites in Cobb 500 chicks. At the 7th day of incubation, a total of 250 eggs were randomly divided into five groups with 5 replications of 10 eggs of each: negative control (normal eggs), positive control (0.5 mL saline), 1.5 mg Astragalus membranaceus, 3.0 mg Astragalus membranaceus, and 4.5 mg in ovo Astragalus membranaceus injection. Live body weight and gain were not significantly (linear and quadratic) affected by in ovo injection of Astragalus membranaceus. Kidney and liver functions were influenced by in ovo injection of Astragalus membranaceus polysaccharides in broiler chickens. Antioxidant enzymes were quadratically increased with Astragalus membranaceus polysaccharides, and the highest values achieved with 4.5 mg. The MDA concentration was linearly and quadratically decreased with in ovo injection of Astragalus membranaceus polysaccharides when compared to negative control. The highest values of IgG and IgM were achieved with 1.5 mg Astragalus membranaceus polysaccharides when compared to all other groups. In conclusion, our results indicate that in ovo injection of Astragalus membranaceus polysaccharides 1.5-4.5 mg in broiler eggs significantly improved serum ALT, AST, AP, creatinine enzymes, antioxidant activity, and immune function.

Quercetin Alleviates the Immunotoxic Impact Mediated by Oxidative Stress and Inflammation Induced by Doxorubicin Exposure in Rats.

Doxorubicin (DOX) is a chemotherapeutic agent against hematogenous and solid tumors with undesirable side effects including immunosuppression. Quercetin (QUR), a natural flavonoid abundant in fruits and vegetables, has a potent antioxidant activity. The aim of the current study was to assess the impact of QUR on DOX-induced hematological and immunological dysfunctions in a rodent model. Randomly grouped rats were treated as follows: control, QUR alone (50 mg/kg for 15 days per os), DOX alone (2.5 mg/kg I/P, three times a week, for two weeks), and co-treated rats with QUR for 15 days prior to and concomitantly with DOX (for two weeks), at the doses intended for groups two and three. DOX alone significantly disrupted the erythrogram and leukogram variables. Serum immunoglobulin (IgG, IgM, and IgE) levels and the activities of catalase (CAT) and superoxide dismutase (SOD) in spleen were declined. The DNA damage traits in spleen were elevated with an upregulation of the expression of the apoptotic markers (p53 and Caspase-3 genes) and the proinflammatory cytokines (IL-6 and TNF-α genes), while the expression of CAT gene was downregulated. These biochemical changes were accompanied by morphological changes in the spleen of DOX-treated rats. Co-treatment with QUR abated most of the DOX-mediated alterations in hematological variables, serum immunoglobulins, and spleen antioxidant status, pro-inflammatory and apoptotic responses, and histopathological alterations. In essence, these data suggest that QUR alleviated DOX-induced toxicities on the bone marrow, spleen, and antibody-producing cells. Supplementation of chemotherapy patients with QUR could circumvent the DOX-induced inflammation and immunotoxicity, and thus prevent chemotherapy failure.

Interactive effects of dietary quercetin nanoparticles on growth, flesh antioxidant capacity and transcription of cytokines and Aeromonas hydrophila quorum sensing orchestrating genes in Nile tilapia (Oreochromis niloticus).

Recently, the concept of incorporating natural products into nanocarriers has been intended to promote fish growth and health via modulating their stability and bioavailability. In this concern, the potential role of reformulated quercetin into nanocarriers was examined, for the first time, on Nile tilapia's performance and immunity, flesh quality and antioxidant indices and disease resistance. Five hundred fish assigned into five experimental groups with formulated diets containing quercetin nanoparticles (QT-NPs) at levels of 0, 100, 200, 300 and 400 mg/kg were challenged with Aeromonas hydrophila (A. hydrophila) after 12 weeks feeding trial. Fish final body weight gain and feed efficiency were significantly maximized in groups enriched with 300 and 400 mg/kg of QT-NPs. Significant reduction in total saturated fatty acids and an elevation in polyunsaturated fatty acids' contents were noticed in fish fed higher QT-NPs doses. The levels of Hydrogen peroxide, reactive oxygen species and malondialdehyde, the markers of meat antioxidant capacity, were reduced by higher inclusion levels of QT-NPs. Accordingly, serum activities and transcriptional levels of GSH-Px, CAT and SOD genes were increased with elevated QT-NPs levels. Immune responses mediated by upregulation of IL-10 and TGF-ß and downregulation of IL-1ß, IL-8 and TNF-α mRNA levels were found to be positively affected by QT-NPs. Dietary QT-NPs downregulated the expression of ahyI and ahyR quorum sensing genes conferring protection against A. hydrophila challenge. This study concluded that supplementation of quercetin in encapsulated nanoparticles could improve its efficacy making it as a compelling approach to improve fish performance and as a promising drug candidate against A. hydrophila virulence.

Performance, Serum Biochemical and Immunological Parameters, and Digestive Enzyme and Intestinal Barrier-Related Gene Expression of Broiler Chickens Fed Fermented Fava Bean By-Products as a Substitute for Conventional Feed.

Improving the nutritional quality of unconventional feed ingredients such as fava bean by-products can enhance their utilization by broiler chickens. Hence, the quality of fermented fava bean by-products (FFB), in addition to growth, nutrient digestibility, digestive enzyme, and intestinal barrier-related gene expression, and serum biochemical and immunological parameters were evaluated in response to different levels of FFB. A total of 500 1-day-old broiler chicks (46.00 ± 0.388 g) were allocated to five groups with 10 replicates each (100 chicks per treatment). The first group was fed a corn-soybean diet (control diet), and the other four groups were fed a diet containing 5, 15, 25, and 35% FFB for 38 days. Birds fed 25% FFB exhibited maximum body weight gain (increase by 12.5%, compared with the control group) and the most improved feed conversion ratio. Additionally, birds fed FFB at 15, 25, and 35% showed improved dry matter and crude protein digestibility. Moreover, birds fed FFB at 25 and 35% exhibited a decrease in ileal pH and an increase in fiber digestibility (p < 0.05). Upregulation of digestive enzyme genes (AMY2A, PNLIP, and CCK) was observed in groups fed with FFB. The most prominent upregulation of genes encoding tight junction proteins (claudin-1, occludin, and junctional adhesion molecules) in the duodenum was observed in chicks fed 25 and 35% FFB (increase of 0.66-, 0.31-, and 1.06-fold and 0.74-, 0.44-, and 0.92-fold, respectively). Additionally, the highest expression level of enterocyte protective genes [glucagon-like peptide (GLP-2), mucin-2 (MUC-2), and fatty acid-binding protein (FABP-6)] was detected in duodenum of chicks fed high levels of FFB. Substitution of corn-soybean diet with FFB had an inhibitory effect on cecal pathogenic microbes (Escherichia coli and Clostridium perfringens) and increased beneficial microflora (Lactobacilli and Bifidobacterium), especially at high levels. Additionally, an increase was observed in IgM and lysozyme activity, with no effect on IgA in all groups fed FFB. All levels of FFB decreased cholesterol levels. Based on our results, we concluded that substitution of corn-soybean diet with FFB can improve the growth rate and nutrient digestibility of broiler chickens, enhance their intestinal barrier functions, and increase the number of beneficial microorganisms. Using FFB at 25% had a positive effect on the growth performance of broiler chickens, and it could be utilized in poultry farms.

Supplementing Garlic Nanohydrogel Optimized Growth, Gastrointestinal Integrity and Economics and Ameliorated Necrotic Enteritis in Broiler Chickens Using a Clostridium perfringens Challenge Model.

Necrotic enteritis (NE) caused by Clostridium perfringens (C. perfringens) results in impaired bird growth performance and increased production costs. Nanotechnology application in the poultry industry to control NE outbreaks is still not completely clarified. Therefore, the efficacy of dietary garlic nano-hydrogel (G-NHG) on broilers growth performance, intestinal integrity, economic returns and its potency to alleviate C. perfringens levels using NE challenge model were addressed. A total of 1200 male broiler chicks (Ross 308) were assigned into six groups; four supplemented with 100, 200, 300 or 400 mg of G-NHG/kg diet and co-challenged with C. perfringens at 21, 22 and 23 d of age and two control groups fed basal diet with or without C. perfringens challenge. Over the total growing period, the 400 mg/kg G-NHG group had the most improved body weight gain and feed conversion efficiency regardless of challenge. Parallel with these results, the mRNA expression of genes encoding digestive enzymes (alpha 2A amylase (AMY2A), pancreatic lipase (PNLIP) and cholecystokinin (CCK)) and intestinal barriers (junctional adhesion molecule-2 (JAM-2), occludin and mucin-2 (Muc-2)) were increased in groups fed G-NHG at higher levels to be nearly similar to those in the unchallenged group. At 14 d post challenge, real-time PCR results revealed that inclusion of G-NHG led to a dose-dependently decrease in the C. perfringens population, thereby decreasing the birds' intestinal lesion score and mortality rates. Using 400 mg/kg of G-NHG remarkably ameliorated the adverse effects of NE caused by C. perfringens challenge, which contributed to better growth performance of challenged birds with rational economic benefits.

Effects of different feeding regimens with protease supplementation on growth, amino acid digestibility, economic efficiency, blood biochemical parameters, and intestinal histology in broiler chickens.

BACKGROUND: This study was conducted to estimate the impacts of using varied feeding regimens with or without protease supplementation on the growth performance, apparent amino acid ileal digestibility (AID%), economic efficiency, intestinal histology, and blood biochemical parameters of broiler chickens. Three hundred one-day-old chicks (Ross 308 broiler) were randomly allotted to a 3 × 2 factorial design. The experimental design consisted of three feeding regimens; FR1: a recommended protein SBM diet, FR2: a low-protein SBM diet, and FR3: a low-protein diet with the inclusion of 5% DDGS and 5% SFM, with or without protease supplementation (250 mg/kg). RESULTS: Increased feed intake and feed conversion ratio were observed in the FR3 treatment during the starter stage and decreased body weight and body weight gain during the grower stage. However, there was no significant effect of the different feeding regimens, protease supplementation, or interaction on the overall performance. The economic value of diets also remained unaffected by the different feeding regimens, protease supplementation, or interaction. Protease supplementation resulted in lowering the AID% of tryptophan and leucine. Reduced AID% of methionine was evident in the FR2 + VE and FR3 - VE treatments. Histological findings substantiated the FR3 treatment mediated a decrease in the duodenal and jejunal villous height (VH), jejunal villous width (VW), and ileal VW, whereas, increase in the ileal crypt depth (CD). The FR2 + VE treatment reduced the VH:CD ratio in the duodenum. The duodenal CD and the jejunal goblet cell count were reduced as a consequence of protease supplementation. The FR3 + VE treatment documented a rise in duodenal CD, while an increase in the jejunal goblet cell count was observed in the FR3 - VE treatment. The FR3 treatment enhanced the IgM serum levels compared to the FR1 and FR2 treatments. IgM serum levels were also elevated following protease supplementation. FR3 + VE treatment increased IgM serum levels. The highest serum ALP was found in the FR3 treatment, whereas the lowest level was obtained in the FR2 treatment. CONCLUSION: Low-protein SBM-based diets could be used without affecting the birds' growth. Altered morphometric measures of the intestine and increased IgM and ALP levels indicated the low-protein SBM/DDGS-SFM diet-induced damage of the intestinal histoarchitecture and immune system of birds. These different diets and protease supplementation failed to affect economic efficiency positively.

Promising Role of Growth Hormone-Boosting Peptide in Regulating the Expression of Muscle-Specific Genes and Related MicroRNAs in Broiler Chickens.

Appropriate skeletal muscle development in poultry is positively related to increasing its meat production. Synthetic peptides with growth hormone-boosting properties can intensify the effects of endogenous growth hormones. However, their effects on the mRNA and miRNA expression profiles that control muscle development post-hatching in broiler chicks is unclear. Thus, we evaluated the possible effects of synthetic growth hormone-boosting peptide (GHBP) inclusion on a chicken's growth rate, skeletal muscle development-related genes and myomiRs, serum biochemical parameters, and myofiber characteristics. A total of 400 one-day-old broiler chicks were divided into four groups supplied with GHBP at the levels of 0, 100, 200 and 300 µg/kg for 7 days post-hatching. The results showed that the highest levels of serum IGF-1 and GH at d 20 and d 38 post-hatching were found in the 200 µg/kg GHBP group. Targeted gene expression analysis in skeletal muscle revealed that the GHBP effect was more prominent at d 20 post-hatching. The maximum muscle development in the 200 µg/kg GHBP group was fostered by the upregulation of IGF-1, mTOR, myoD, and myogenin and the downregulation of myostatin and the Pax-3 and -7 genes compared to the control group. In parallel, muscle-specific myomiR analysis described upregulation of miR-27b and miR-499 and down-regulation of miR-1a, miR-133a, miR-133b, and miR-206 in both the 200 and 300 µg/kg GHBP groups. This was reflected in the weight gain of birds, which was increased by 17.3 and 11.2% in the 200 and 300 µg/kg GHBP groups, respectively, when compared with the control group. Moreover, the maximum improvement in the feed conversion ratio was achieved in the 200 µg/kg GHBP group. The myogenic effects of GHBP were also confirmed via studying myofiber characteristics, wherein the largest myofiber sizes and areas were achieved in the 200 µg/kg GHBP group. Overall, our findings indicated that administration of 200 µg/kg GHBP for broiler chicks could accelerate their muscle development by positively regulating muscle-specific mRNA and myomiR expression and reinforcing myofiber growth.

Synergetic Action of Forskolin and Mevastatin Induce Normalization of Lipids Profile in Dyslipidemic Rats through Adenosine Monophosphate Kinase Upregulation.

In the present study, we examined the synergetic effect of forskolin and mevastatin administration on lipid profile and lipid metabolism in omental adipose tissue in dyslipidemic rats. The study was conducted on forty male albino rats. The rats were randomly classified into four main groups of ten animals in each group as follows: group A, served as control nontreated; group B, rats that received Triton WR 1339 (500 mg/kg); group C, rats that received Triton WR 1339 with forskolin (100% FSK extract 0.5 mg/kg/day) for four weeks; and group D, dyslipidemic rats received both mevastatin and forskolin. At the end of the experimental period, blood and omental adipose tissue samples were collected, preserved, and used for biochemical determination of lipid profile and mRNA expression profile of adenylate cyclase (AC), hormone-sensitive lipase, respectively (HSL), and adenosine monophosphate-activated protein kinase (AMPK). The results showed a significant decline in the serum concentration of total cholesterol, LDL-cholesterol, and triglycerides, although there was a significant increase in serum levels of HDL-cholesterol and glycerol in rats received forskolin alone or with mevastatin when compared with control and dyslipidemic groups. The mRNA expression levels of AC, HSL, and AMPK were significantly increased in omental adipose tissue of rats received forskolin when compared with other groups. In conclusion, forskolin acts synergistically with mevastatin to lower lipid profile and improve lipid metabolism in dyslipidemic rats through upregulation of AMPK expression.

Immune response and susceptibility of Nile tilapia fish to Aeromonas hydrophila infection following the exposure to Bifenthrin and/or supplementation with Petroselinum crispum essential oil.

Bifenthrin (BF) is a widely used 3rd generation type I pyrethroid with a potential toxic effect in fish. Nevertheless, its effect on the immune system remains unclear. In the present study, Oreochromis niloticus was exposed to BF at 0.68 µg/L for 60 days, followed by evaluating the hematological, biochemical, and immunological responses. Additionally, the potential of parsley (Petroselinum crispum) essential oil (PEO) to ameliorate the BF-induced toxic insults was explored. Our data have shown reductions in the growth performance with alterations observed in the hematological variables, protein profile and serum biomarkers of stress. DNA oxidative damage was evidenced by elevation of serum 8-hydroxy-2-deoxyguanosine (8-OHdG) content. BF-exposed fish presented also decline in serum lysozyme activity and levels of immunoglobulins (IgG and IgM) and nitric oxide (NO), with diminished resistance to Aeromonas hydrophila challenge. Furthermore, the RT-PCR analysis showed an upregulated expression pattern of immune -related genes including interleukin 1ß (IL-1ß), interferon - γ (IFN-γ) and tumor necrosis factor - α (TNF-α) genes in the liver tissue. Dietary co-supplementation of PEO at 1 or 2 mL/kg diet with concomitant BF exposure, alleviated the adverse effects of the insecticide in a dose-dependent manner. The observations from this study demonstrate the immunomodulation by BF and provide further insight into the protective properties of PEO and strengthen its applicability as a promising feed supplement to fish.

Adsorptivity of mercury on magnetite nano-particles and their influences on growth, economical, hemato-biochemical, histological parameters and bioaccumulation in Nile tilapia (Oreochromis niloticus).

Among toxic pollutants, Mercury (Hg) is a toxic heavy metal that induces harmful impacts on aquatic ecosystems directly and human being's health indirectly. This study confirmed the in vitro magnetic potential of magnetite Nano-Particles (Fe3O4 NPs) against waterborne Hg exposure-induced toxicity in Nile tilapia (Oreochromis niloticus). We further evaluate the safety profile of Fe3O4 NPs on fish growth, hemato-biochemical, histological parameters, bioaccumulation in muscles, and economy. Magnetite nanoparticles were characterized, adsorption loading to Hg ions was investigated, and testing different concentrations of Fe3O4 NPs (0.2, 0.4, 0.6, 0.8, and 1.0 mg/L) was applied to determine the highest concentration of adsorption. An in vivo experiment includes 120 fish with an average weight of 26.2 ± 0.26 g were randomly divided into 4 equal groups, each group had three replicates (n = 30 fish/group; 10 fish/ replicate). All groups were fed on a reference basal diet and the experiment was conducted for 30 days. The first group (G1) was allocated as a control. The second group (G2) received 1.0 mg/L aqueous suspension of Fe3O4 NPs. The third group (G3) was exposed to an aqueous solution of Hg ions at a concentration of 0.025 mg/L. Meanwhile, the fourth group (G4) acquired an aqueous suspension composed of a mixture of Hg ions and Fe3O4 NPs as previously mentioned. Throughout the exposure period, the clinical signs, symptoms, and mortalities were recorded. The Hg ions-exposed group induced the following consequences; reduced appetite resulting in reduced growth and less economic efficiency; microcytic hypochromic anemia, leukocytosis, lymphopenia, and neutrophilia; sharp and clear depletion in the immune indicators including lysozymes activity, immunoglobulin M (IgM), and Myeloperoxidase activities (MPO); significant higher levels of ALT, AST, urea, creatinine, and Superoxide dismutase (SOD); histological alterations of gill, hepatic and muscular tissues with strong expression of apoptotic marker (caspase 3); and a higher accumulation of Hg ions in the muscles. Surprisingly, Fe3O4 NPs-supplemented groups exhibited strong adsorption capacity against the Hg ions and mostly removed the Hg ions accumulation in the muscles. Also, the hematological, biochemical, and histological parameters were recovered. Thus, in order to assess the antitoxic role of Fe3O4 NPs against Hg and their safety on O. niloticus, and fill the gap of the research, the current context was investigated to evaluate the promising role of Fe3O4 NPs to prevent Hg-exposure-induced toxicity and protection of fish health, which ascertains essentiality for sustainable development of nanotechnology in the aquatic environment.

Growth, Hemato-Biochemical Parameters, Body Composition, and Myostatin Gene Expression of Clarias gariepinus Fed by Replacing Fishmeal with Plant Protein.

In this study, we evaluated the consequences of replacement of fishmeal with plant protein sources (soybean and sunflower meal) on fish growth parameters, haemato-biochemical factors, body composition, and myostatin gene expression of Clarias gariepinus. A total of 150 C. gariepinus were organized in glass aquaria into five investigational groups, with each group in triplicate (30 fish/group; 10 fish/replicate). Group 1 was fed a control diet (15% fishmeal). The other groups were fed diets where fishmeal was replaced gradually with plant protein, with 10% fishmeal in the second group (D1), 7.5% fishmeal in the third group (D2), 5% fishmeal in the fourth group (D3), and 0% fishmeal in the fifth group (D4). There were no significant differences regarding growth performances and body composition among the groups, except that the feed conversion ratio was improved in D4. The different diet types did not affect hematologic parameters and blood indices. Serum growth hormone and amylase levels also revealed no significant (p = 0.09 and 0.55, respectively) differences among the groups, while serum lipase levels decreased significantly (p = 0.000) due to partial (D2) or complete (D4) substitution of fishmeal with plant protein. The replacement of fishmeal had no effects on liver (p = 0.51) and kidney functions (p = 0.34). However, D4 showed the best profit and economic efficiency compared to the other groups. Altogether, we concluded that substitution of fishmeal with plant protein sources is economically beneficial and may be without any adverse effects on growth parameters, body composition, or hematologic and biochemical parameters, but with the addition of synthetic amino acids.

Thymol nanoemulsion promoted broiler chicken's growth, gastrointestinal barrier and bacterial community and conferred protection against Salmonella Typhimurium.

The present study involved in vivo evaluation of the growth promoting effects of thymol and thymol nanoemulsion and their protection against Salmonella Typhimurium infection in broilers. One-day old 2400 chicks were randomly divided into eight groups; negative and positive control groups fed basal diet without additives and thymol and thymol nanoemulsion groups (0.25, 0.5 and 1% each). At d 23, all chicks except negative control were challenged with S. Typhimurium. Over the total growing period, birds fed 1% thymol nanoemulsion showed better growth performance even after S. Typhimurium challenge, which came parallel with upregulation of digestive enzyme genes (AMY2A, PNLIP and CCK). Additionally, higher levels of thymol nanoemulsion upregulated the expression of MUC-2, FABP2, IL-10, IgA and tight junction proteins genes and downregulated IL-2 and IL-6 genes expression. Moreover, 1% thymol nanoemulsion, and to lesser extent 0.5% thymol nanoemulsion and 1% thymol, corrected the histological alterations of cecum and liver postinfection. Finally, supplementation of 1% thymol, 0.5 and 1% thymol nanoemulsion led to increased Lactobacilli counts and decreased S. Typhimurium populations and downregulated invA gene expression postinfection. This first report of supplying thymol nanoemulsion in broiler diets proved that 1% nano-thymol is a potential growth promoting and antibacterial agent.

Preventive effect of artemisinin extract against cholestasis induced via lithocholic acid exposure.

Obstructive cholestasis characterized by biliary pressure increase leading to leakage of bile back that causes liver injury. The present study aims to evaluate the effects of artemisinin in obstructive cholestasis in mice. The present study was carried out on 40 adult healthy mice that were divided into 4 groups, 10 mice each; the negative control group didn't receive any medication. The normal group was fed normally with 100 mg/kg of artemisinin extract orally. The cholestatic group fed on 1% lithocholic acid (LCA) mixed into control diet and cholestatic group co-treated with 100 mg/kg of artemisinin extract orally. Mice were treated for 1 month then killed at end of the experiment. A significant increase in alanine aminotransferase, aspartate aminotransferase, and total and direct bilirubin was detected in mice exposed to LCA toxicity. That increase was significantly reduced to normal values in mice co-treated with artemisinin. LCA toxicity causes multiple areas of necrosis of irregular distribution. However, artemisinin co-treatment showed normal hepatic architecture. Moreover, LCA causes down-regulation of hepatic mRNA expressions of a set of genes that are responsible for ATP binding cassette and anions permeability as ATP-binding cassette sub-family G member 8, organic anion-transporting polypeptide, and multidrug resistance-associated protein 2 genes that were ameliorated by artemisinin administration. Similarly, LCA toxicity significantly down-regulated hepatic mRNA expression of constitutive androstane receptor, OATP4, and farnesoid x receptor genes. However, artemisinin treatment showed a reasonable prevention. In conclusion, the current study strikingly revealed that artemisinin treatment can prevent severe hepatotoxicity and cholestasis that led via LCA exposure.

Chronic effects of soft drink consumption on the health state of Wistar rats: A biochemical, genetic and histopathological study.

The present study was performed to examine the effects of chronic soft drink consumption (SDC) on oxidative stress, biochemical alterations, gene biomarkers and histopathology of bone, liver and kidney. Free drinking water of adult male Wistar rats was substituted with three different soft drinks: Coca­Cola, Pepsi and 7­Up, for three consecutive months. The serum and organs were collected for examining the biochemical parameters associated with bone, liver and kidney functions. Semi­quantitative reverse transcription polymerase chain reaction was used to observe the changes in the expression of genes in the liver and kidney, which are associated with oxidative stress resistance. Histopathological investigations were performed to determine the changes in bone, liver and kidney tissues using hematoxylin and eosin stains. SDC affected liver, kidney and bone function biomarkers. Soft drinks increased oxidative stress, which is represented by an increase in malondialdehyde and a decrease in antioxidant levels. SDC affected serum mineral levels, particularly calcium and phosphorus. Soft drinks downregulated the expression levels of glutathione­S­transferase and super oxide dismutase in the liver compared with that of control rats. Rats administered Coca­Cola exhibited a hepatic decrease in the mRNA expression of α2­macroglobulin compared with rats administered Pepsi and 7­Up. On the other hand, SDC increased the mRNA expression of α1­acid glycoprotein. The present renal studies revealed that Coca­Cola increased the mRNA expression levels of desmin, angiotensinogen and angiotensinogen receptor compared with the other groups, together with mild congestion in renal histopathology. Deleterious histopathological changes were reported predominantly in the bone and liver of the Coca­Cola and Pepsi groups. In conclusion, a very strict caution must be considered with SDC due to the increase in oxidative stress biomarkers and disruption in the expression of certain genes associated with the bio­vital function of both the liver and kidney.

Origanum Majoranum Extract Modulates Gene Expression, Hepatic and Renal Changes in a Rat Model of Type 2 Diabetes.

The present study was conducted to test the effect of Origanum MajoranumExtract (OME) of leaves on alterations induced in a model of type 2 diabetic rats. Adult male Wistar rats were fed high fat diet for 3 weeks and injected a single dose of streptozotocin (35 mg/kg) intraperitoneally to induce type 2 diabetic rats. Diabetic rats were given aqueous extract of OME in a dose of 20 mg/kg orally for 3 weeks. Changes in lipid profiles, glucose, insulin, expression of some genes related to glucose metabolism and histopathological changes in liver and kidney were examined. Administration of OME improved and normalized dyslipidemia recorded in type 2 diabetic rats together with reduction in glucose and insulin levels. OME induced up-regulation in gene expression of glucose [adiponectin and glucose transporter-2 (GLUT-2)] and lipid metabolism [lipoprotein lipase (LPL)]. Moreover, OME normalized histopathological changes occurred in liver and kidney of diabetic rats. OME decreased lipids accumulation in liver and kidney and increased regeneration of hepatic parenchyma and restored normal renal architecture with disappearance of fat droplets. In conclusion, OME improved dyslipidemia associated with type 2 diabetes through regulation of genes related to glucose and lipid metabolism.