l-Arginine deiminase (ADI) has a powerful anticancer activity against various tumors, via arginine depletion, arresting the cell cycle at G1 phase. However, the current clinically tried bacterial ADI displayed a higher antigenicity and lower thermal stability. Thus, our objective was to purify and characterize this enzyme from thermophilic fungi, to explore its catalytic and antigenic properties for therapeutic uses. ADI was purified from thermophilic Aspergillus fumigatus KJ434941 to its electrophoretic homogeneity by 5.1-fold, with molecular subunit 50 kDa. The purified ADI was PEGylated and covalently immobilized on dextran to explore its catalytic properties. The specific activity of free ADI, PEG-ADI, and Dex-ADI was 26.7, 21.5, and 18.0 U/mg, respectively. At 50°C, PEG-ADI displays twofold resistance to thermal denaturation (t1/2 13.9 h), than free ADI (t1/2 6.9 h), while at 70°C, the thermal stability of PEG-ADI was increased by 1.7-fold, with similar stability to Dex-ADI with the free one. Kinetically, free ADI had the higher catalytic affinity to arginine, followed by PEG-ADI and Dex-ADI. Upon proteolysis for 30 min, the residual activity of native ADI, PEG-ADI, and Dex-AD was 8.0, 32.0, and 20.0% for proteinase K and 10.0, 52.0, and 90.0% for acid protease, respectively. The anticancer activity of the ADIs was assessed against HCT, HEP-G2, and MCF7, in vitro. The free and PEG-ADI exhibits a similar cytotoxic efficacy for the tested cells, lower than Dex-ADI. The free ADI had IC50 value 22.0, 16.6, and 13.9 U/mL, while Dex-ADI had 3.98, 5.18, and 4.43 U/mL for HCT, MCF7, and HEPG-2, respectively. The in vitro anticancer activity of ADI against HCT, MCF7, and HEPG-2 was increased by five-, three-, and threefold upon covalent modification by dextran. The biochemical and hematological parameters of the experimented animals were not affected by ADIs dosing, with no signs of anti-ADI immunoglobulins in vivo. The in vivo half-life time of free ADI, PEG-ADI, and Dex-ADI was 29.7, 91.1, 59.6 h, respectively. The present findings explored a novel thermostable, less antigenic ADI from thermophilic A. fumigatus, with further molecular and crystallographic analyses, this enzyme will be a powerful candidate for clinical trials.

The aim of the present study was to evaluate mannan oligosaccharides (MOS) or glycerol (GLY) as a carbon source on biofloc systems of Nile tilapia (O. niloticus) juveniles. Fish (n = 750) were reared in open flow (Controls) or biofloc systems (B-GLY and B-MOS) fed with a plant or fish protein source over a period of twelve weeks. Total ammonia nitrogen and nitrate decreased in the biofloc groups, while biofloc volume increased in B-MOS. Compared to the controls, B-MOS and B-GLY exhibited higher weight gain and improved feed conversion, irrespectively of the diet. Serum level of C-reactive protein was reduced, while IgM and lysozyme activity was higher in the B-MOS fish, compared to other groups. Intestinal Bacillus spp. count was increased, whereas Vibrio, Aeromonas and Pseudomonas spp. counts decreased in B-MOS reared groups, compared to the other groups. The proinflammatory cytokine (IL-8 and IFN-γ) transcript expression was upregulated in B-MOS more than B-GLY reared groups. Compared to the controls, the virulence of Aeromonas hydrophila was decreased in the B-MOS and B-GLY groups. The results indicate several benefits of using MOS as a carbon source in a biofloc Nile tilapia system; a cost benefit analysis is required to assess the economic viability of this.

This study was conducted to compare the effects of commercially available (C) and green synthesized (GS) Zinc oxide nanoparticles (ZnO-NPs) on immunological responses of common carp (Cyprinus carpio) skin mucus. GS ZnO-NPs were generated using Thymus pubescent and characterized by UV–vis diffuse reflectance spectroscopy (DRS), Fourier-transform infrared spectroscopy (FTIR), X-ray powder diffraction (XRD), scanning electron microscope (SEM), and energy-dispersive X-ray spectroscopy (EDX). Fish (n = 150) were randomly allocated into five groups in triplicate and received a waterborne concentration of 0% (control), 25%, and 50% of LC50 96 h of commercially available (C1 and C2) and green synthesized ZnO-NPs (GS1 and GS2) for 21 days. Results from XRD displayed ZnO-NPs with 58 nm in size and UV-vis DRS, EDX, and FT-IR analysis showed that some functional groups from plant extract bonded to the surface of NPs. The SEM images showed that ZnO-NPs have conical morphology. Acute toxicity study showed a higher dose of LC5096h for green synthesized ZnO-NPs (78.9 mg.L−1) compared to the commercial source (59.95 mg.L−1). The highest activity of lysozyme and alternative complement activity (ACH50) were found in control and GS1 groups. A significant decrease in alkaline phosphatase activity (ALP) was found in C1 and C2 groups compared to other treatments. Protease activity (P) was significantly decreased in the C2 group compared to the control and GS groups. Total immunoglobulin (total Ig) content was the highest in the control. In addition, total Ig in the GS1 group was higher than GS2. The exposure to ZnO-NPs lowered total protein content in all experimental groups when compared to control. Present findings revealed lower induced immunosuppressive effects by green synthesized ZnO-NPs on key parameters of fish skin mucus.

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.

No abstract

Taxol, a diterpenoid was initially isolated from the bark of Taxus brevifolia, approved by FDA in 1994 as a powerful drug for metastatic ovarian carcinoma, breast and lung cancer. However, due to limitations in the production of this drug based on this plant source, the productive potentiality of fungi of this compound opened a new avenue for its commercial production. In this study, among the twenty fungal isolates screened for Taxol production, Aspergillus terreus had the highest potentiality to produce Taxol (131.2 µg/ml). The productivity of Taxol by A. terreus has been maximized by nutritional optimization using inhibitors and growth regulators. The yield of Taxol by A. terreus was maximally obtained (0.663µg/ ml) by growing the fungal isolate on potato dextrose broth medium, amended with addition of biotin at 150 µg/ml for 20 days. The chemical structure Taxol extracted of A. terreus has been verified by proton and carbon NMR, IR and UV analyses. The activity of A. terreus Taxol has been assessed towards multiple cell lines, displaying a strong anticancer activity. In conclusion, the productivity of Taxol by A. terreus has been greatly improved upon using biotin as chemical modulator, that open a new avenue for commercializing the Taxol yield by fungi.

Camel meat is one of the most consumed meats in Arab countries. The use of natural antimicrobial agents to extend the shelf life of fresh camel meat, control Campylobacter jejuni contamination, and preserve meat quality is preferred. In this study, we determined the antimicrobial effects of using 1% or 2% Citrox alone or in combination with 1% chitosan on the survival of C. jejuni in vitro and on camel meat samples during storage at 4 or 10 °C for 30 days in vacuum packaging. We determined the total viable count (TVC (cfu/g)), total volatile base nitrogen (TVB-N) content, and pH of the treated camel meat samples every three days during storage. The shelf lives of camel meat samples treated with 2% Citrox alone or in combination with 1% chitosan were longer than those of camel meat samples treated with 1% Citrox alone or in combination with 1% chitosan at both the 4 and 10 °C storage temperatures, with TVCs of <100 cfu/g after the first ten days and six days of storage at 4 and 10 °C, respectively. The addition of Citrox (1% and 2%) and 1% chitosan to camel meat samples and the application of vacuum storage were more effective than using Citrox (1% and 2%) alone and led to a reduction in C. jejuni in approximately 4.0 and 3.5 log cycles at 4 and 10 °C, respectively. The experimental results demonstrated that using a Citrox-chitosan combination improved the quality of camel meat and enhanced the long-term preservation of fresh meat for up to or more than 30 days at 4 °C.

The Pseudomonas putida strain was primarily identified and tested in vitro against antibiotic sensitivity for several antibiotics using the disc diffusion method. This isolate was also tested against sensitivity to carvacrol oil (c) and formic acid (f). The genotyping of Pseudomonas spp. and virulotyping for P. putida isolate was carried out and verified by 16S rDNA-PCR amplification. Furthermore, we assessed the efficacy of carvacrol oil and formic acid in vivo for treatment of P. Putida infection. For the in vivo challenge, 180 fish (Nile tilapia, Oreochromis niloticus) were divided into six groups: (G1: control (unchallenged), G2: carvacrol prophylaxis (3 g/kg), G3: formic acid prophylaxis (5 mL/kg), G4: control positive (challenged), G5: carvacrol treatment (3 g/kg), and G6: formic acid treatment (5 mL/kg); 30 fish per group) with three replicates. Following the challenge, nitric oxide and lysozyme activity were measured as essential indicators for fish immunity. The antioxidant parameters including SOD and catalase were computed to reflect the antioxidant status. Furthermore, relative percent survival (RPS) and mortality percent were evaluated to indicate functional immunity. The findings of the antibiotic sensitivity test showed that ciprofloxacin exhibited the largest inhibition zone. Additionally, formic acid (f) displayed the greatest inhibition zone compared to carvacrol oil (c) and was more effective in stimulating the immune-antioxidant response compared to carvacrol oil. The tested exotoxin A (tox A), exoenzyme S (exo S), and the nan1 associated-virulence genes were identified in the P. putida isolate. Overall, the current study verified the virulence of P. putida and highlighted the promising role of dietary addition of formic acid for enhancing the immune-antioxidant indicators and for mitigating P. putida infection. Future studies could be devoted to this field.