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In medicine, silver nanoparticles (AgNPs) are employed often. They do, however, have negative impacts, particularly on the reproductive organs. This research aimed to assess AgNP impact on the testis and the possible intracellular mechanisms to induce testicular deteriorations in rats at various concentrations and different time intervals. Sprague Dawley rats (n = 40) were allocated into four equal groups: the control one, and three other groups injected intra-peritoneally with AgNP solution 0.25, 0.5, and 1 mg/kg b.w. respectively for 15 and 30 days. Our findings revealed that AgNPs reduced body and testicular weights, estradiol (E2) and testosterone (T) hormone levels, and sperm parameters while elevating the nitric oxide and malondialdehyde levels with inhibition of reduced glutathione contents in testicular tissue. Interestingly, AgNPs significantly upregulated the testicular inducible nitric oxide synthase, B cell lymphoma 2 (Bcl-2)-associated X, transforming growth factor, and alpha-smooth muscle actin (α-SMA) expression levels. However, apurinic/apyrimidinic endo deoxyribonuclease 1 (APE1), NAD (P) H quinone dehydrogenase 1 (NQO1), and Bcl-2 expression levels were all downregulated indicating exhaustion of body antioxidant and repairing defense mechanisms in testicles in comparison with the control rats. Various histological alterations were also detected which dramatically increased in rats sacrificed after 30 days such as loss of the lining cells of seminiferous tubules with no spermatozoa and tubular irregularities associated with thickening of their basement membranes. Immunolabeling implicated in the apoptotic pathway revealed a negative expression of Bcl-2 and marked immunoreactivity for caspase-3 after 30 days of AgNP treatment in comparison to the control rats. To our knowledge, there have been no previous publications on the role of the α-SMA, APE1, and NQO1 genes in the molecular pathogenesis of AgNP testicular cytotoxicity following AgNP acute and chronic exposure. Graphical Abstract

Recently, the high cost of aquafeed affected fish farming feasibility in some countries, including Egypt. The imported soybean meal and corn ingredients consume a large amount of the hard currency, thereby increasing feed prices. Thus, the current study investigated the different sources of soybean and corn on the performances of Nile tilapia. Fish fed with the diet I (based on Egyptian soybean meal cultivar and cornmeal cultivar) or diet II (based on imported soybean meal cultivar and cornmeal cultivar) in a 90-day feeding trial. The results showed no marked effects on the growth performance, protein efficacy ratio, and FCR in the case of fish-fed diet I or diet II. No histological alterations were observed in the skeletal muscle, hepatopancreas, spleen, and intestines, while the diet I-fed group showed normal architecture of the above-listed organs. The expression of liver and muscle IGF-1 showed no changes in fish-fed diet I or diet II. No diet-related variations were observed in IL-1β expression in the spleen but increased regulation in the liver of the diet II group compared to the diet I group. Furthermore, significant upregulation of SOD and HSP70 genes was seen in the spleen and liver of the diet II-fed group. We conclude that the inclusion of the Egyptian soybean meal cultivar and cornmeal cultivar (diet I) did not reduce the growth performance and immune-related genes compared with the imported soybean meal cultivar and cornmeal cultivar (diet II).

Ochratoxin A (OTA) is one of the most dangerous and that pollute agricultural products, inducing a variety of toxic effects in humans and animals. The current study explored the protective effect of different concentrations of Aspergillus awamori (A. awamori) against OTA (0.3 mg/kg diet) induced renal and cardiac damage by exploring its mechanism of action in 60 New Zealand white male rabbits. Dietary supplementation of A. awamori at the selected doses of 50, 100, and 150 mg/kg diet, respectively, for 2 months significantly improved the rabbit’s growth performance; modulated the suppressed immune response and restored the altered hematological parameters; reduced the elevated levels of renal injury biomarkers such as urea, creatinine, and alkaline phosphatase; and increased serum total proteins concentrations. Moreover, it also declined enzymatic activities of cardiac injury biomarkers, including AST, LDH, and CK-MB. A. awamori alleviated OTA-induced degenerative and necrotic changes in the kidney and heart of rabbits. Interestingly, A. awamori upregulated Nrf2/OH-1 signaling pathway. Therefore enhanced TAC, CAT, and SOD enzyme activities and reduced OTA-induced oxidative and nitrosative stress by declining iNOS gene expression and consequently lowered MDA and NO levels. In addition to attenuating renal and cardiac inflammation via reducing IL-1β, TNF-α gene expressions in a dose-dependent response. In conclusion,this is the first report to pinpoint that dietary incorporation of A. awamori counteracted OTA-induced renal and cardiac damage by potentiating the rabbit’s antioxidant defense system through its potent antioxidant, free radical scavenging, and anti-inflammatory properties in a dose-dependent response. Based on our observations, A. awamori could be utilized as a natural protective agent against ochratoxicosis in rabbits. Graphical abstract

This study focuses on the relationship between myostatin (MyoS), myogenin (MyoG), and the growth hormone/insulin-like growth factor-1 (GH/IGF-1) axis for muscle growth and histopathological changes in muscle after an Aeromonas hydrophila infection. A total number of 90 Nile tilapia (55.85 g) were randomly allocated into two equal groups of three replicates each. The first group was an uninfected control group that was injected intraperitoneally (ip) with 0.2 ml phosphate buffer saline (PBS), while the second group was injected ip with 0.2 ml (1.3 × 10

The trials of finding non-conventional and alternative aquafeed ingredients are increasing. In this sense, this study evaluated the influence of coconut oil on the growth, feed utilization, immune, and antioxidative responses of Nile tilapia. Five test diets were formulated by mixing coconut oil with the other ingredients at 0, 1, 2, 3, and 4% of the total ration and presented for tilapia for 60 successive days. The final weight, SGR, weight gain (WG), and feed intake were superior in fish delivered 2% of coconut oil (P < 0.05). Concurrently, fish that received 2% coconut oil had lower FCR and higher PER than fish of the control and 4% groups (P < 0.05). Higher lipase activity was observed in fish of 2% and 3% levels than the remaining groups (P < 0.05). Besides, the amylase and protease activities of fish in 1%, 2%, and 3% groups were higher than the 0% level (P < 0.05). The total blood cholesterol, RBCs, and PCV showed higher values in Nile tilapia fed 2% and 3% coconut oil (P < 0.05). The lysozyme and phagocytic activities were higher in fish fed 2% and 3% levels than the control (P < 0.05), while the phagocytic index in 2% and 3% levels was higher than 0% and 4% levels. Furthermore, SOD and CAT were higher in fish fed 1%, 2%, and 3% than fish fed 0% and 4% levels while GSH was higher in fish of 1%, 2%, and 3% than fish fed 0% level (P < 0.05). However, the MDA level was markedly lower in fish fed 25, 3%, and 4% coconut oil than the 0% level (P < 0.05). The intestine's histological structure in all groups appeared normal, forming of intestinal villi projecting from the intestinal wall. Also, the structure of the hepatopancreas had a normal architecture in all groups. To sum up, the inclusion of coconut oil at 2 to 3% is recommended as a replacer for fish oil in Nile tilapia diets.

Chlorpyrifos (CPF) is an insecticide that is commonly applied in the agriculture sector. However, little is known about the protective role of Spirulina platensis (SP) and/or β-glucan (BG) on African catfish exposed to chronic CPF toxicity. The fish (95 ± 5 g, initial weight) were assigned to 5 fiberglass tanks (500 L, 50 fish/tank) where the 1st and 2nd fed the basal diet, while the 3rd, 4th, and 5th fed diets with SP, BG, and SP+BG at 0.5%, respectively. Fish in 2nd, 3rd, 4th, and 5th groups were exposed to CPF at a dose of 1.5 mg/L and fed the respective diets for 60 days. In comparison with the control group, CPF-exposed fish exhibited significantly lower (P ≤ 0.05) body weights, feed intake, red blood cells count, hemoglobin concentration, packed cell volume (PCV) (%), lymphocytes, monocytes, phagocytic activity, and phagocytic index, while feed conversion ratio, white blood cell count, and neutrophils count were significantly increased. Fish exposed to CPF also revealed a significant elevation in aspartate aminotransferase (AST), alanine aminotransferase (ALT), cholesterol, triglycerides, low-density lipoproteins (LDL), very-low-density lipoproteins (vLDL), glucose concentration, urea, and creatinine as well as low total proteins, albumin, globulins, and high-density lipoprotein (HDL) concentration. Fish exposed to CPF also exhibited a high concentration of malondialdehyde while glutathione content, glutathione peroxidase, and catalase activities were significantly decreased in the liver, gills, brain, and intestine tissues. Moreover, exposure to CPF resulted in higher transcription of cytochrome P450 (CYP1A-P450) gene expression than the 1st group. Histopathological investigations revealed various degrees of pathological lesions in different organs like the liver, kidney, brain, spleen, and intestine tissues. Interestingly, dietary SP supplementation either alone or combined with BG significantly ameliorated the alterations mitigated by CPF-induced organ injuries and genotoxicity. Therefore, it could be concluded that SP or/and BG are able to induce the protective consequences on health status, immunity, and antioxidative response of African catfish exposed to CPF.

The current study investigated how different fasting and refeeding regimes would impact Nile tilapia growth performance, histopathological examination, and gene expression of myostatin, myogenin, GH, IGF-1, and NPYa. Nile tilapia fish (n = 120) were randomly allocated into four groups, including the control group fed on a basal diet for 6 weeks (F6), group A starved for 1 week and then refed for 5 weeks (S1F5), group B starved for 2 weeks and then refed for 4 weeks (S2F4), while group C starved for 4 weeks and then refed for 2 weeks (S4F2). Fasting provoked a decrease in body weight coincided with more extended starvation periods. Also, it induced muscle and liver histological alterations; the severity was correlated with the length of fasting periods. Gene expression levels of GH, MSTN, MYOG, and NPYa were significantly increased, while IGF1 was markedly depressed in fasted fish compared to the control group. Interestingly, refeeding after well-planned short fasting period (S1F5) modulated the histopathological alterations. To some extent, these changes were restored after refeeding. Restored IGF-I and opposing fasting expression profiles of the genes mentioned above thus recovered weights almost like the control group and achieved satisfactory growth compensation. Conversely, refeeding following more extended fasting periods failed to restore body weight. In conclusion, refeeding after fasting can induce a compensatory response. Still, the restoration capacity is dependent on the length of fasting and refeeding periods through exhibiting differential morphological structure and expressions pattern for muscle and growth-related genes. Graphical abstract

Despite the extraordinary use of silver nanoparticles (AgNPs) in medicinal purposes and the food industry, there is rising worry about potential hazards to human health and the environment. The existing study aims to assess the hepatotoxic effects of different dosages of AgNPs by evaluating hematobiochemical parameters, oxidative stress, liver morphological alterations, immunohistochemical staining, and gene expression to clarify the mechanism of AgNPs’ hepatic toxic potential. Forty male Sprague Dawley rats were randomly assigned into control and three AgNPs intraperitoneally treated groups 0.25, 0.5, and 1 mg/kg b.w. daily for 15 and 30 days. AgNP exposure reduced body weight, caused haematological abnormalities, and enhanced hepatic oxidative and nitrosative stress with depletion of the hepatic GSH level. Serum hepatic injury biomarkers with pathological hepatic lesions where cholangiopathy emerges as the main hepatic alteration in a dosage- and duration-dependent manner were also elevated. Furthermore, immunohistochemical labelling of apoptotic markers demonstrated that Bcl-2 was significantly downregulated while caspase-3 was significantly upregulated. In conclusion, the hepatotoxic impact of AgNPs may be regulated by two mechanisms, implying the apoptotic/antiapoptotic pathway via raising BAX and inhibiting Bcl-2 expression levels in a dose-dependent manner. The TGF-β1 and α-SMA pathway which triggered fibrosis with incorporation of iNOS which consequently activates the inflammatory process were also elevated. To our knowledge, there has been no prior report on the experimental administration of AgNPs in three different dosages for short and long durations in rats with the assessment of Bcl-2, BAX, iNOS, TGF-β1, and α-SMA gene expressions. Graphical abstract

The trials of finding non-conventional and alternative aquafeed ingredients are increasing. In this sense, this study evaluated the influence of coconut oil on the growth, feed utilization, immune, and antioxidative responses of Nile tilapia. Five test diets were formulated by mixing coconut oil with the other ingredients at 0, 1, 2, 3, and 4% of the total ration and presented for tilapia for 60 successive days. The final weight, SGR, weight gain (WG), and feed intake were superior in fish delivered 2% of coconut oil (P0.05). Concurrently, fish that received 2% coconut oil had lower FCR and higher PER than fish of the control and 4% groups (P0.05). Higher lipase activity was observed in fish of 2% and 3% levels than the remaining groups (P0.05). Besides, the amylase and protease activities of fish in 1%, 2%, and 3% groups were higher than the 0% level (P0.05). The total blood cholesterol, RBCs, and PCV showed higher values in Nile tilapia fed 2% and 3% coconut oil (P0.05). The lysozyme and phagocytic activities were higher in fish fed 2% and 3% levels than the control (P0.05), while the phagocytic index in 2% and 3% levels was higher than 0% and 4% levels. Furthermore, SOD and CAT were higher in fish fed 1%, 2%, and 3% than fish fed 0% and 4% levels while GSH was higher in fish of 1%, 2%, and 3% than fish fed 0% level (P0.05). However, the MDA level was markedly lower in fish fed 25, 3%, and 4% coconut oil than the 0% level (P0.05). The intestine's histological structure in all groups appeared normal, forming of intestinal villi projecting from the intestinal wall. Also, the structure of the hepatopancreas had a normal architecture in all groups. To sum up, the inclusion of coconut oil at 2 to 3% is recommended as a replacer for fish oil in Nile tilapia diets.

Paracetamol is extensively consumed as an analgesic and antipyretic drug, but at a high dose level, it leads to deleterious side effects, such as hepatic and nephrotoxicity. This research aimed to estimate the prophylactic efficacy of Chlorella vulgaris and/or thiamine against paracetamol (P) induced hepatorenal and cardiac toxicity. Forty-eight female Wistar rats were randomly divided into eight equal groups (n = 6 rats). Group 1, normal control group. Group 2, Paracetamol group. Groups 3, 4 and 5 were treated with Silymarin drug, Chlorella vulgaris alga, Chlorella vulgaris alga supplemented with thiamine, respectively daily for 7 successive days, then all were administered Paracetamol (2gm/kg. bwt.). While, Groups 6, 7 and 8 were treated by Silymarin, Chlorella vulgaris alga, Chlorella vulgaris supplemented with thiamine, respectively daily for 7 successive days without paracetamol administration. Our results clarified that Paracetamol toxicity caused significant adverse effects on hematological, serum biochemical parameters, and oxidant -antioxidant status as well as histopathological picture of heart, liver, and kidney. However, in the Paracetamol intoxicated groups pretreatment either with Chlorella vulgaris alone or plus thiamine successfully improved the undesirable deleterious effects of paracetamol, and restored almost all variables to near their control levels. This study has finished to that oxidative stress participates in the pathogenesis of paracetamol-induced toxicity in rats and using Chlorella vulgaris alga either alone or plus thiamine alongside their health benefits can protect against oxidative harmful effects induced by paracetamol through their free radical scavenging and powerful antioxidant effects, and they can be used as propylactic agents against paracetamol-induced toxicity.

Blue-green microalga Spirulina platensis (SP) gained more attention for its antioxidant and/or anti-inflammatory properties magnifying its beneficial effects as a feed additive and for cosmetic and biomedical applications. This study was performed to examine the impact of SP on the cutaneous wound and burn healing and to develop an understanding of the correlation between the sequelae of wound healing and the molecular expression patterns of wound healing-related genes as angiogenic basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) and fibrosis-related genes as transforming growth factor-β (TGF-β) and α-smooth muscle actin (α-SMA) in rat wound models. To achieve these goals, two experiments were performed on 32 Wister male rats that were divided into 4 groups of 8 rats each. Each experiment was represented by 2 groups; the control group (CG) and the Spirulina group (SG). A full-thickness wound (1.5 × 1.5 cm) and burn wound (2 × 2 cm) were made on the back of each generally anaesthetized rat and the areas of wound and burn were measured on days of 0, 3, 6, 9, 12, and 15 and 0, 3, 6, 9, 12, 15, 18, and 21 post-wound and post-burn respectively. In both experiments, SP was topically applied on the backs of wounded and burned rats in Spirulina treated groups. The phases of wound granulation tissues were detected histopathologically. Immunohistochemistry was used to determine the expressions of (TGF-B1) and (VEGF). Furthermore, the relative quantification of gene expression was implemented using the (bFGF), (VEGF), (TGF-Ɓ1), and (α-SMA) as target genes. Histopathological examination revealed inflammatory cell infiltration, angiogenesis, epithelialization, and extracellular matrix deposition and wound contraction in SG as compared to CG in both experiments. Immunohistochemistry results showed a significant improvement in the VEGF and TGF-β1 expression levels of SG in both experiments. Interestingly, SG in both experiments revealed upregulation of angiogenic genes (bFGF and VEGF) and downregulation of fibrotic genes (TGF-β1 and α-SMA). In conclusion, our findings suggest that the topically applied Spirulina promoted wound healing. Thus, SP can be used as a biomedical application to treat various skin wounds and may reveal a potential molecular basis for future promising antifibrotic agents against scar formation.

The current study aimed to evaluate whether clenbuterol, a β2-adrenergic agonist, supplementation in Nile tilapia (Oreochromis niloticus) diets can influence growth and blood parameters. Besides, assessment of adipogenic genes as fatty acid synthase (FAS) and lipoprotein lipase (LPL) which is a key enzyme in the regulation of the flux of fatty acids in liver, muscle, and adipose tissue as well as muscle growth-regulating genes as myostatin (MYO) in muscle and insulin-like growth factor-1 (IGF-1) in liver. The fish were allocated into three equal groups; control group that fed basal diet only and the other two groups fed a basal diet containing clenbuterol at two doses 5 ppm and 10 ppm/kg diet for 30 consecutive days. Results revealed that clenbuterol supplementation significantly increased body weight, decreased liver, spleen and abdominal fat weights, and decreased total circulatory cholesterol and triacylglycerol levels. Moreover, clenbuterol inhibits lipogenesis by downregulation of FAS gene expression by dose and time-dependent manner in the liver while enhanced lipolysis in both the liver and in the adipose tissue. Moreover, lipolysis was reduced in muscle by dose 10 ppm on day 30. Furthermore, clenbuterol presented higher gene expression of MYO and IGF-1 in muscle and liver respectively by dose 5 ppm at day 15 on the other hand, these findings were reversed by day 30 compared with control. In conclusion, clenbuterol efficacy was apparent in a dose and time response pattern to boost growth and reduce fat deposition rates, indicating for the first time that clenbuterol has a profitable growth impact on Nile tilapia.

Wound healing is a public health concern. Licorice gained a great attention for its antioxidant and anti-inflammatory properties which expand its valuable effects as a herbal medicine. In this study, we pointed out to the wound healing potential and the mechanism by which licorice alcoholic extract can modulate cutaneous wound healing through immune, antioxidant, histopathological, immunohistochemical (IHC) and molecular studies. 24 Wister rats were assigned into 3 groups (n = 8 each); control group, topical and oral supplied groups. Licorice extract administration significantly increased total and differential leucocyte counts, phagocytic activity of neutrophils, antioxidant biomarkers as superoxide dismutase (SOD), glutathione peroxidase activities (GPx) and reduced glutathione (GSH) content with a notable reduction in oxidative stress marker malondialdehyde (MDA). Moreover, histopathological findings detected complete re-epithelialization with increasing collagen synthesis while IHC results revealed a significant enhancement in the expression of α-SMA, PDGFR-α, FGFR1 and Cytokeratin 14 in licorice treated groups compared with the control group. Licorice extract supplementation accelerated wound healing by increasing angiogenesis and collagen deposition through up-regulation of bFGF, VEGF and TGF-β gene expression levels compared with the control group. UPLC-PDA-MS/MS aided to authenticate the studied Glycyrrihza species and recognized 101 potential constituents that may be responsible for licorice-exhibited potentials. Based on our observations we concluded that licorice enhanced cutaneous wound healing via its free radical-scavenging potential, potent antioxidant activities, and anti-inflammatory actions. Therefore, licorice could be used as a potential alternative therapy for wound injury which could overcome the associated limitations of modern therapeutic products.

This study aimed to explore some fertility biomarkers in seminal plasma (SP) and blood of rams with different field fertility, based on their conception and lambing rates. Ejaculates were collected from fertile (n = 20) and subfertile (n = 10) Ossimi rams by electroejaculation for 16 replicates. Each ejaculate was analyzed for volume and sperm characteristics before centrifugation at 12,000 × g for 30 min at 4 °C to collect SP. Blood samples were collected from all rams and centrifuged to collect serum which was frozen stored at – 30 °C until analysis. Seminal and serum levels of superoxide dismutase, nitric oxide, testosterone, thyroxin and malondialdehyde as well as seminal fructose and serum glucose were determined. Ovine SP proteins were extracted and separated by gel electrophoresis. Both ejaculate volume (P

The present trial investigated the effects of dietary fenugreek on the histopathology, oxidative status, and immune-related gene expression in Nile tilapia (Oreochromis niloticus) (15 ± 0.5 g) infected with Aeromonas hydrophila. Fish were fed two diets with or without fenugreek supplementation at 3%. The trial was lasted for 8 weeks then fish were divided into four groups; the first and fourth groups were intra-peritoneally injected with phosphate buffer saline (PBS) while the second and third groups were challenged with sublethal dose of A. hydrophila. At 6 h, 24 h and 1-week post challenge, several serum immune and immune-related gene expressions and antioxidant defense parameters as well as the histopathological alterations were evaluated. The results indicated that administration of fenugreek supplemented diets could significantly increase the growth performance parameters and decreased the feed conversion ratio as compared to the non-supplemented groups (P

Antioxidants have valuable effects on the process of spermatogenesis, particularly with diabetes mellitus (DM). Therefore, the present study investigated the impact and the intracellular mechanisms by which thymoquinone (TQ) works against diabetes-induced testicular deteriorations in rats. Wistar male rats (n = 60) were randomly allocated into four groups; Control, Diabetic (streptozotocin (STZ)-treated rats where diabetes was induced by intraperitoneal injection of STZ, 65 mg/kg), Diabetic + TQ (diabetic rats treated with TQ (50 mg/kg) orally once daily), and TQ (non-diabetic rats treated with TQ) for 12 weeks. Results revealed that TQ significantly improved the sperm parameters with a reduction in nitric oxide (NO) and malondialdehyde (MDA) levels in testicular tissue. Also, it increased testicular reduced glutathione (GSH) levels and superoxide dismutase (SOD) activity. Interestingly, TQ induced downregulation of testicular inducible nitric oxide synthase (iNOS) and nuclear factor kappa-B (NF-κB) and significantly upregulated the aromatase protein expression levels in testicles in comparison with the diabetic rats. In conclusion, TQ treatment exerted a protective effect against reproductive dysfunction induced by diabetes not only through its powerful antioxidant and hypoglycemic effects but also through its downregulation of testicular iNOS and NF-κB along with upregulation of aromatase expression levels in diabetic rats.