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The present research amid to investigate the toxic effect of diazinon (DZN) and assess the modulatory role of chitosan nanoparticle (ChNP), orange peel extract (OPE), spirulina platensis (SP) and curcumin nanoparticles (CNP) as feed additives in O. niloticus fish. A total of 900 fish were divided into six equal groups, each with 3 replicates by random distribution. The 1st group was kept as the control group which was fed a basal diet. The 2nd group (DZN) was given a basic diet and exposed to 1/10th of the 96-hour LC50 of DZN. The 3rd group (DZN+ChNP) was exposed to DZN (0.28 mg/L) + 1 gm ChNP/kg diet. The 4th group (DZN+OPE) was exposed to DZN (0.28 mg/L) + 2 gm OPE/kg diet. The 5th group (DZN+SP) was exposed to DZN (0.28 mg/L) + 5 gm SP/kg diet, while, the 6th group (DZN+SP) was exposed to DZN (0.28 mg/L) + 150 mg CNP/kg diet. Apart from feed conversion, DZN decreased fish's growth performance parameters as compared to the control, while these values were modulated with ChNP, SP or CNP supplement to fish with DZN exposure. Fish exposed to DZN had the lowest values Hb, RBCs WBCs, total protein, albumin and globulin and the highest values of ALT, AST and uric acid while dietary supplementation with ChNP, OPE, SP and CNP to DZN exposed fish restored their levels to the control values. Fish group fed diet supplemented with DZN+OPE or DZN+ChNP recorded the lowest value of ALT among all studied groups. DZN altered the lipid profile of exposed fish while the DZN+SP group had the best values of all lipid profile among all groups followed by DZN+CNP group. Fish exposed to DZN had lower serum lysozyme activity, immunoglobulin (M, A and G) levels and antioxidant parameters (SOD, GSH, CAT and TAC) while dietary supplementation of ChNP or OPE to DZN exposed fish had higher values for these traits but still under control values. The highest level of lipid peroxidation marker malondialdehyde (MDA) was observed in DZN group while dietary supplementation of ChNP or OPE to DZN exposed fish decreased the MDA level. According to the genetic distance and dendrogram for the analysis of genetic diversity between treatments, compared to control; it is clear that the highest genetic diversity was found with DZN group samples (4.80) while dietary treatments with ChNP, OPE or SP to DZN exposed fish showed the lowest diversity of (3.00, 3.32 and 3.87), respectively. It could be concluded that long-term exposure to a sub-lethal DZN dose resulted in growth retardation, haemato-biochemical alteration, immune depression, oxidative stress and genetic material damage of Nile tilapia. Furthermore, the antioxidant and protective properties of ChNP, OPE, SP, and CNP induced nearly total protection in fish exposed to DZN through diet.

SUMMARY: This study evaluated the effects of pumpkin oil (PKO), a novel feed additive, on growth, carcass, digestive enzymes, blood metabolites, and cecal bacterial load in broiler chickens. 250 one-wk-old chicks were randomly assigned into 5 equal groups. Group 1(control) was fed the basal diet (BD). Groups 2, 3, 4, and 5 were fed the basal diet (BD) with 0.50, 1.00, 1.50, and 2.00 ml of PKO/kg diet, respectively. According to GC-MS analysis, pumpkin oil contains Anthracene, Pyrene, Anethole, Estragole, Oleic Acid, Eugenol, 1-Dodecanamine, N, N-dimethyl- and Hexadecanoic acid, etc. The broilers fed the PKO-supplemented diet had better growth and feed conversion ratio (FCR) than those fed the control diet, with no effect on feed intake (P > 0.05). The highest body weight gain (P < 0.05) was obtained with 1 and 2 ml of PKO/kg diet compared to 0.5 and 1.5 ml/kg. The best FCR (P < 0.05) was achieved with 1 ml of PKO/kg diet compared to other treatments. Ileal lipase, amylase, and protease levels were increased significantly while the reverse was observed for plasma aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine, urea, and uric acid (P < 0.05) by PKO-supplemented diets. PKO supplementation did not, however, alter the percentage of giblets, internal organs, dressing, or carcass (P > 0.05). When compared to control birds, broilers' plasma level of high-density lipoprotein (HDL) was higher and those of triglycerides (TG), total cholesterol (TC), very low-density lipoprotein (VLDL), and low-density lipoprotein (LDL) were lower at every tested level of PKO (P < 0.05). Similarly, all PKO levels increased Lactobacilli and decreased the caecal counts of Salmonella, coliforms, and E. coli (P < 0.05). Total antioxidant capacity (TAC) and superoxide dismutase (SOD) were significantly (P < 0.05) increased while the malondialdehyde (MDA) was decreased in broilers fed PKO-supplemented diet vs control birds. PKO enhanced immune response parameters including total protein, globulin, IgA, IgG, and C3 (P < 0.05) dose dependently. It was concluded that adding PKO in the broilers’ diet positively affected growth performance, induced hypolipidemic effect, enhanced immunity, and antimicrobial and antioxidant activities.

Recently, nanomaterials including alumina have been extensively involved in aquaculture for many purposes resulting in potential hazards in fish. However, its effect on the developing metabolic organs and lipid metabolism remains unclear. This study, investigated the effects of aluminium nanoparticles (AL-NPs) on the metabolic organs of the growing zebrafish larvae including intestine, liver, and pancreas in addition to the potential ameliorative role of rosmarinic acid (RA) against AL-NPs - induced toxicity. Briefly, fertilized zebrafish eggs were divided into 4 groups. The 1st group was kept as a control. The 2nd group was exposed to RA (0.5 mg/L). The 3rd group was exposed to AL-NPs (160 mg/L). The 4th group received both RA and AL-NPs at the same concentrations from 1st day post fertilization (dpf) till the 14th dpf. Here, AL-NPs exposed group showed the highest mortality rate, external morphology defects and gastrointestinal malformations after larval-to-juvenile transition. Interestingly, mixing RA to AL-NPs significantly reduced these effects. Additionally, AL-NPs induced intestinal epithelium defects at the histological and genetic levels and abnormal lipid retention via up-regulation of the lipogenesis-related genes and down-regulation of the hepatic fatty acid β-oxidation. However, addition of RA to AL-NPs could reverse these effects. AL-NPs also induce inflammatory response and mediated endoplasmic reticulum ER and cellular oxidative stress indicated by qRT-PCR and western blot analysis which were then be reduced by RA. Collectively, AL-NPs were highly correlated with the induction of steatohepatitis and lipogenesis after the larval-to-juvenile transition and this could be relief in the presence of RA suggesting it as a promising nutrient for fish.

Necrotic enteritis caused by Clostridium perfringens (C. perfringens) is characterized by poor performance and higher mortality rates in poultry farms. Novel dietary intervention involving bioactive molecules loaded into smart magnetized nano-system with a potent antioxidant function (quercetin-loaded Fe3O4-NPs), was evaluated for their impact on growth performance, intestinal immune and antioxidant defenses, and resistance against Clostridium perfringens in a necrotic enteritis challenge model. Four experimental groups comprising a total of 200 one-day-old Ross 308 broiler chickens were fed different diets: a control basal diet, a diet supplemented with quercetin (300 mg/kg), a diet with Fe3O4-NPs (60 mg/kg), and a diet with quercetin-loaded Fe3O4-NPs (300 mg/kg). These groups were then challenged with C. perfringens during the grower period. Dietary inclusion of quercetin-loaded Fe3O4-NPs prominently reduced C. perfringens colonization and its associated virulence genes expression, which subsequently restored the impaired growth performance and intestinal histopathological changes in challenged broilers. Quercetin-loaded Fe3O4-NPs supplemented group displayed higher Lactobacillus and Bifidobacterium counts, upregulation of intestinal host defense antimicrobial peptides related genes (avian β-defensin 6 and 12) and downregulation of intestinal inflammatory regulated genes (Interleukin-1 beta, C-X-C motif chemokine ligand 8, tumor necrosis factor-α, chemokine C–C motif ligand 20, inducible nitric oxide synthase and cycloox-ygenase-2). Intestinal redox balance was boosted via upregulation of catalase, superoxide dismutase, glutathione peroxidase and heme Oxygenase 1 genes along with simultaneous decrease in hydrogen peroxide, reactive oxygen species and malondialdehyde contents in groups fed quercetin-loaded Fe3O4-NPs. Overall, new nutritional intervention with quercetin-loaded Fe3O4-NPs impacted better immune and antioxidant defenses, attenuated C. perfringens induced necrotic enteritis and contributed to better performance in the challenged birds.

ABSTRACT: This study evaluated the effects of dietary butyric acid (BA) on the Japanese quail' performance, immunology, lipid profile, cecal microbiota, and antioxidant levels. 250 unsexed, one-week-old quail chicks were divided into 5 groups, each with fifty chicks (5 replicates of 10 chicks). The first group was given the basal diet (BD), while the 2nd to 5th groups were fed BD with 50, 100, 150, and 200 mg BA/kg, respectively. The results indicated that BA improved weight gain and FCR (p < 0.05) and decreased total FI. The 200 mg BA/kg of diet showed the lowest FI (p < 0.05) and the best FCR (p > 0.05). BA boosted immunity through increasing IgA, IgM, IgG, and Complement 3. Significantly lower alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH) were observed at 150 and 200 mg BA/kg (P < 0.05) than the control group. The BA-supplemented quail showed lower total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), and very low-density lipoprotein (VLDL) than the control one. This effect was more pronounced for 100 and 200 mg of BA/kg. However, high low-density lipoprotein (HDL) did not differ from the control group (p > 0.05). BA at ≥100 mg/kg diet reduced malondialdehyde (MDA) and induced greater levels of superoxide dismutase (SOD), total antioxidant capacity (TAC), glutathione peroxidase (GPX), globulin, total protein, digestive enzymes than the control group (P < 0.05). BA decreased cecal E. coli, Salmonella, Enterococcus, and Coliforms and increased Lactic acid bacteria (p < 0.05) compared to non-supplemented group. Collectively, the inclusion of 100 mg BA/kg diet is ideal for Japanese quail production and health.

Abstract 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.

Abstract 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.

Nonylphenol (NP) is a common toxicant that constitutes a potential hazard to the aquatic system. This study assessed the protective role of Salvia officinalis leaves powder (SOLP) against NP toxicity. We addressed mortalities, growth retardation, hepato-renal dysfunction, immune-antioxidant suppression, histopathological degeneration, and gene down-regulation induced by NP-toxicity. And the potential activity of SOLP against these negative impacts in Mirror carp (Cyprinus carpio var specularis). Acute toxicity assay (static 96-h) was primarily performed to investigate the 96-h LC50 of NP in C. carpio and it was recorded at 1.01 mg/L reckoned by Finney probit analysis. Fish (N = 240) were assigned to 4 treatments for 2 months. The first (control) and the second (SOLP) groups were fed on basal diets that were enriched with 0.0 and 4 g/kg of diet SOLP without NP exposure. The third (NP) and fourth (SOLP+NP) groups were provided with the same diets plus exposed to 1/10 of the 96 h-LC50 of NP (0.1 mg/L). The outcomes revealed that NP induced higher mortalities (46), significantly (p

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-1β (IL-1β), 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-1β, 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.

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, β-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-1β, 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.

Silica nanoparticles (SiNPs) are a new class of nanomaterials with unique properties due to their nanoscale size. Currently, SiNPs constitute a potential value in aquaculture; nonetheless, misuse induces toxicity to fisheries and ecosystems. Hence, the present prospective is a pioneer study that illustrates the acute toxicity of SiNPs to African catfish, Clarias gariepinus (B.) and the behavioral response in a renewal static system. Moreover, the influence of SiNPs on neuro-related, stress, immune-related, and oxidative stress indices was investigated. Fish groups of average body weights (73.09 ± 2.20 g) were exposed to 96-h of various concentrations of SiNPs including 0, 50, 100, 150, 200, 250, and 300 mg L−1 without water exchange. The value of 96-h LC50 of SiNPs was computed using Finney's probit analysis. Here we show that the exposure of fish to SiNPs resulted in remarkable ethological alterations such as lethargic, surfacing, inactive, slow swimming manner, and nervousness. By increasing the SiNPs concentration, the behavior scores and mortalities reached the upper limit of top values. The comparison with the control revealed a significant dose-dependent effect by increasing in the activity of 8-hydroxy-2-deoxyguanosine (8-OHdG) and malondialdehyde (MDA) with a concomitant elevation in the stress-controlled parameters (cortisol and nor-epinephrin) of all the fish groups undergone the SiNPs exposure. In contrast, there was a clear reduction in the acetylcholine esterase (AChE) level, immunological biomarkers (lysozymes and nitric oxide), and antioxidant enzymes (glutathione peroxidase (GPX) and superoxide dismutase (SOD)) of the exposed groups and the reduction elevated by increasing the SiNPs exposure concentration in comparison to the control group. Based on the study outcomes, the 96-h LC50 of SiNPs recorded 144.54 mg L−1 in C. gariepinus. We anticipate our assay to be a starting point for further applying the one-tenth of 96-h LC50- as an alternative safe treatment in therapy after considering the acute toxicity to preserve the aquaculture production and sustain human health. Additionally, it was approved that, the toxicity by the exposure to SiNPs observed clear alterations in fish behavior, neurological activity, immune and oxidative responses reflecting physiological disturbance.

The experiment investigated the effect of dietary orange pulp (OP) with or without enzymes supplementation on the performance parameters, chemical composition, serum biochemistry, and some liver gene expression in tilapia for twelve weeks. The studied fish (n = 300) were allotted into six groups according to OP level (0%, 10%, and 20%) with or without enzymes supplementation (+/-). The performance results indicated that the inclusion level of OP by 20% negatively affected FCR and decreased final body weight. The dietary enzymes did not alleviate the unfavorable effect of OP on the final body weight. Higher dietary replacement levels of OP reduced serum lipid profile, serum proteins, and carcass fat, but increased carcass protein and ash compared to the control. The dietary enzymes produced higher serum lipids and proteins as compared to non-supplemented groups. Fish-fed the control diet supplemented with enzymes (G2) expressed higher RNA levels of GHR and IGF1 genes compared to the other groups. Excess dietary OP lowered the RNA level of FAS and LPL genes but improved simultaneously when supplemented with enzymes. Overall, the higher dietary OP (20%) with or without enzymes supplementation, did not benefit the tilapia performance, metabolic, and genetic indices, and the chemical composition of the carcass. But, the lower level of OP (10%) supplemented with enzymes could be used in tilapia diets without adverse effects on performance, health, and carcass composition.

The present study assessed the protective aspects of the dietary exposure to black cumin (Nigella sativa, NS) powder on immune response, liver and kidney enzymes, tissue antioxidant capacity, gene expression, and testing the antibacterial activity against Burkholderia cepacia (B. cepacia, BC). Nile tilapia (O. niloticus) (n = 150) were divided into 5 groups, the first was control (Cont) neither challenged nor treated, and fed a basal diet for 59 days. The second one received 30 g kg−1 of NS-incorporated diet and farmed for 45 days (45NS). The third (Cont-BC) and the fourth (14NS-BC) groups were fed a basal diet, while the fifth (59NS-BC) was fed NS-supplemented diet for 45 days. Then, the last three groups were intraperitoneally inoculated with 0.2 mL of virulent BC, and kept under observation for 14 days. During the observation period, Cont-BC group was continuously received a basal diet, while 14NS-BC and 59NS-BC were fed on NS-supplemented diet. Results showed that fish received NS-dietary supplement exhibited significantly (P

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.

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 1β (IL-1β), 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.

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.

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.

In modern aquaculture, enriching Nile tilapia’s diet with omega-3 poly-unsaturated fatty acids (PUFAs) not only plays an important role in its general health but also fortifies its fillet with omega-3-PUFAs. However, the major challenge affecting their delivery is their high instability due to oxidative deterioration. Thus, the prospective incorporation of omega-3-PUFAs into nanocarriers can enhance their stability and bioactivity. In this regard, the effect of reformulated omega-3-NPs was investigated on Nile tilapia’s performance, flesh antioxidant stability, immunity, and disease resistance. Four fish groups supplemented with omega-3-PUFAs-loaded nanoparticles (omega-3 NPs) at levels of 0, 1, 2, and 3 g/kg diet and at the end of feeding trial fish challenged with Aeromonas hydrophila. Fish performance (weight gain and feed conversion) was improved in groups supplemented with omega-3-NPs (2 and 3 g/kg diet). The deposition of omega-3-PUFAs in fish flesh elevated with increasing dietary omega-3-NPs. Simultaneously the oxidative markers (H2O2, MDA, and reactive oxygen species) in fish flesh were reduced, especially with higher omega-3-NPs. Post-challenge, downregulation of IL-1β, IL-6, IL-8, TNF-α, and caspase-1 were noticed after dietary supplementation of omega-3-NPs. Moreover, mRNA expression of autophagy-related genes was upregulated while the mTOR gene was downregulated with higher omega-3 NPs levels. Lower expression of A. hydrophila ahyI and ahyR genes were detected with omega-3 NPs supplementation. In conclusion, omega-3-NPs application can fortify tilapia flesh with omega-3-PUFAs and augment its performance, immunity, and disease resistance against Aeromonas hydrophila.

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.

The objectives of the current study were first to determine the prevalence of non O157:H7E. coli, especially Shiga toxin-producing Escherichia coli (STEC) in retailed milk and dairy products in Egypt. Second, the antimicrobial resistance profiling and virulence genes of the isolated E. coli strains were screened. Third, evaluation of the inhibitory effects of D-tryptophan against E. coli O26:H11 was further performed. The results revealed that 20% (30/150) of the samples were STEC positive, with 64 isolates harboring some virulent genes, such as Stx1, Stx2, eaeA, and hylA. Serological identification revealed four different pathotypes belonging to EPEC, ETEC, EHEC, and EIEC. Antimicrobial susceptibility testing revealed that 100%, 98.44%, 92.19%, 71.87%, 65.63% and 64.06% of the isolates had a resistance against tetracycline, oxacillin, erythromycin, nalidixic acid, sulphamethoxazol, and ampicillin, respectively. D-tryptophan addition (40 mM) to E. coli O26:H11-inoculated soft cheese and ice cream revealed a significant reduction (p < 0.05) in bacterial growth, especially when accompanied with other food stressors. D-Tryptophan is considered as an effective food preservative and as a promising alternative candidate in the dairy industry.

Optimal combinations of essential oils (EOs) can enhance performance and maintain poultry productivity. The effects of EOs with black pepper oil (BPO) or radish seed oil (RSO) on performance and the expression of digestive enzymes, lipogenesis, immunity, and autophagy-related genes in broiler chickens were explored. Six dietary treatments for 300 one-day-old chicks were allocated as follows: controls were fed a basal diet, one group was fed an EO-supplemented diet (1.5 g/kg diet of parsley, mint, and carrot seed oils (1:1:1)), and other groups received Eos + BPO0.25, Eos + BPO0.5, Eos + RSO0.25, and Eos + RSO0.5 treatments, with a basal diet containing EOs plus BPO or RSO at the level of 0.25 or 0.5 g/kg, respectively. Supplementation with 0.5 g/kg of EOs plus BPO or RSO resulted in the most improved maximum BWG and FCR in broiler chickens. The lactobacilli population was increased in Eos + BPO0.5, followed by EOs + RSO0.5, unlike in the control. The highest expression of the CCK and PNLIP genes was identified in the Eos + BPO group. The FAS and ACC genes were upregulated, while the IgA and IL-10 genes were downregulated, with EOs plus RSO or BPO. The group that received Eos + BPO0.5, followed by Eos + RSO0.5, displayed patterns of higher expression for atg5, atg7, and atg12, with lower expression of mTOR. In summary, a new combination of EOs with 0.5 g/kg BPO had potential growth-promoting and immune-boosting effects in broiler chickens.

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.

This study recommends Carbopol/zinc oxide (ZnO) hybrid nanoparticles gel as an efficient antibacterial agent against different bacterial species. To this end, ZnO nanoparticles were synthesized using chemical precipitation derived from a zinc acetate solution with ammonium hydroxide as its precipitating agent under the effect of ultrasonic radiation. The synthesized ZnO nanoparticles were stabilized simultaneously in a freshly prepared Carbopol gel at a pH of 7. The chemical composition, phase identification, particle size and shape, surface charge, pore size distribution, and the BET surface area of the ZnO nanoparticles, as well as the Carbopol/ZnO hybrid Nanoparticles gel, were by XRD, SEM, TEM, AFM, DLS, Zeta potential and BET instruments. The results revealed that the synthesized ZnO nanoparticles were well-dispersed in the Carbopol gel network, and have a wurtzite-crystalline phase of spherical shape. Moreover, the Carbopol/ZnO hybrid nanoparticles gel exhibited a particle size distribution between ~9 and ~93 nm, and a surface area of 54.26 m2/g. The synthesized Carbopol/ZnO hybrid nanoparticles gel underwent an antibacterial sensitivity test against gram-negative K. pneumonia (ATCC 13883), Bacillus subtilis (ATCC 6633), and gram-positive Staphylococcus aureus (ATCC 6538) bacterial strains, and were compared with ampicillin as a reference antibiotic agent. The obtained results demonstrated that the synthesized Carbopol/ZnO hybrid nanoparticles gel exhibited a compatible bioactivity against the different strains of bacteria.

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.

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.

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.