Composition of bacterial and archaeal communities in the rumen of dromedary camel using cDNA-amplicon sequencing.

The camel is known to survive in harsh environmental conditions, due to its higher digestive efficiency of high-fiber diets compared with other ruminants. However, limited data are available on the microbial community in the rumen of a camel. In this study, the Illumina sequencing of V4 region of 16S rRNA genes based on RNA isolation was employed to get insight into the bacterial and archaeal communities associated with liquid and solid rumen fractions in eight camels under different feeding systems. Camels in group C1 were fed Egyptian clover hay plus concentrates mixture and camels of group C2 were fed fresh Egyptian clover. The results showed that liquid fraction has higher operational taxonomic units (OTUs) than solid fraction, and camel group C1 showed a higher microbial diversity than C2. The UniFrac analysis indicated that the microbial communities in camel groups are distinct. Moreover, phylum Firmicutes and Bacteroidetes dominated the bacterial community and Candidatus Methanomethylophilus dominated the archaeal community with a significant difference in the relative abundance between camel groups. Dominant bacterial genera were Prevotella, Fibrobacteres, Ruminococcus, and Butyrivibrio. There were many negative and positive correlations between and within bacterial and archaeal genera. The composition of microbial community in the rumen of a camel is similar to other ruminants with differences in the abundance.

Community structure and fibrolytic activities of anaerobic rumen fungi in dromedary camels.

Anaerobic fungi colonize the rumen and degrade cellulose and hemicellulose, which enable them to be key players in the lignocellulose fermentation. Consequently, an expansion of knowledge about rumen fungi could increase animal productivity, utilization of lignified forages like alfalfa hay, and enhance fibrolytic enzymes production. Here, we used an Internal Transcribed Spacer 1 (ITS1) clone library to investigate the anaerobic rumen fungi in camel and to investigate their ability to produce cellulase and xylanase in vitro. Rumen fluid was collected from camels fed Egyptian clover (n = 14), and wheat straw (n = 7) and fecal samples were collected from camels fed wheat straw and concentrates (n = 5), or natural grazing plants (n = 10). Neocallimastix and Cyllamyces were the most abundant anaerobic fungi in all camel groups. An anaerobic rumen fungi media containing alfalfa hay as a carbon source was inoculated by rumen and fecal samples to assess the ability of anaerobic rumen fungi in camel gut to produce cellulase and xylanase. The anaerobic gut fungi in the camel is diverse and has cellulolytic and xylanolytic activities, fungal culture from rumen samples of camel fed wheat straw (R2) exhibited highest cellulase production. In addition, many of the sequences in the current study have no equivalent cultured representative, indicating a novel diversity within the camel gut.

Therapeutic enhancement of newly derived bacteriocins against Giardia lamblia.

Trials for identifying efficient anti-giardial agents are still ongoing. Nowadays, bacteriocins have attracted the attention as potential antimicrobial compounds. For the first time, the current study evaluated the therapeutic efficacy of bacteriocins derived from newly isolated Egyptian strains of probiotics Lactobacilli; L. acidophilus (P106) and L. plantarum (P164) against Giardia lamblia. Bacteriocins' efficacy was evaluated both in vitro; by growth inhibition and adherence assays, and in vivo; through estimation of parasite density, intestinal histopathological examination and ultrastructural analysis of Giardia trophozoites. In vivo bacteriocins' clinical safety was assessed. In vitro results proved that 50 µg of L. acidophilus bacteriocin induced reduction of the mean Giardia lamblia trophozoites by 58.3 ± 4.04%, while at lower concentrations of 10 and 20 µg of both L. acidophilus and L. plantarum, non significant reduction of the mean parasite density was achieved. In vitro trophozoites adherence was susceptible to the tested bacteriocins at all studied concentrations with variable degrees, while the highest adherence reduction was demonstrated using 50 µg of L acidophilus bacteriocin. In vivo, oral inoculation of 50 µg/mouse L. acidophilus bacteriocin for 5 successive days resulted in a noteworthy decline of the intestinal parasite density, along with amelioration of intestinal pathology of infected mice. Ultrastructural examination proved thatfive doses of L. acidophilus bacteriocin showed marked changes in cellular architecture of the trophozoites with evident disorganization of the cell membrane, adhesive disc and cytoplasmic components. This is the first reported study of the safe anti-giardial efficacy of L. acidophilus (P106) derived bacteriocin, hence highlighting its great promise as a potential therapeutic safe alternative to existing commercial drugs.

A sustainable green-approach for biofabrication of chitosan nanoparticles, optimization, characterization, its antifungal activity against phytopathogenic Fusarium culmorum and antitumor activity.

Chitosan is a natural non-toxic, biocompatible, biodegradable, and mucoadhesive polymer. It also has a broad spectrum of applications such as agriculture, medical fields, cosmetics and food industries. In this investigation, chitosan nanoparticles were produced by an aqueous extract of Cympopogon citratus leaves as a reducing agent. According to the SEM and TEM micrographs, CNPs had a spherical shape, and size ranging from 8.08 to 12.01 nm. CNPs have a positively charged surface with a Zeta potential of + 26 mV. The crystalline feature of CNPs is determined by X-ray diffraction. There are many functional groups, including C꞊C, CH2-OH, C-O, C-S, N-H, CN, CH and OH were detected by FTIR analysis. As shown by the thermogravimetric study, CNPs have a high thermal stability. For the optimization of the green synthesis of CNPs, a Face centered central composite design (FCCCD) with 30 trials was used. The maximum yield of CNPs (13.99 mg CNPs/mL) was produced with chitosan concentration 1.5%, pH 4.5 at 40 °C, and incubation period of 30 min. The antifungal activity of CNPs was evaluated against phytopathogenic fungus; Fusarium culmorum. A 100% rate of mycelial growth inhibition was gained by the application of 20 mg CNPs/mL. The antitumor activity of the green synthesized CNPs was examined using 6 different cell lines, the viability of the cells reduced when the concentration of green synthesized CNPs increased, the IC50 dose of the green synthesized CNPs on the examined cell lines HePG-2, MCF-7, HCT-116, PC-3, Hela and WI-38 was 36.25 ± 2.3, 31.21 ± 2.2, 67.45 ± 3.5, 56.30 ± 3.3, 44.62 ± 2.6 and 74.90 ± 3.8; respectively.

Application of edible nanocomposites from chitosan/fenugreek seed mucilage/selenium nanoparticles for protecting lemon from green mold.

Green (Penicillium digitatum) mold can severely endanger the citrus fruits production and quality. Targeting the protection of lemon fruits from green mold infestations with nanobiotechnology approach, the fenugreek seed mucilage (FM) was extracted and exploited for biosynthesis of selenium (SeNPs) nanoparticles; their nanocomposites (NCs) with chitosan (CT) was constructed and employed as antifungal materials and edible coating (ECs) to protect lemon fruits against green mold. The nanoparticles formation and conjugations were verified by infrared (FTIR) analysis and electron microscopy. The FM-synthesized SeNPs had particles average of 8.35 nm, were the NCs of them with CT had size mean of 49.33 nm and charged with +22.8 mV. The CT/FM/SeNPs composite exhibited superior antifungal actions toward P. digitatum isolates, up to 32.2 mm inhibition diameter and 12.5 mg/mL inhibitory concentration, which exceeded the actions of imazilil. The microscopic screening of exposed P. digitatum to NCs clarified their mycelial destructive action within 30 h. The coating of infected lemons with fabricated NCs led to complete elimination of green mold development after 10 days of coating, without any infestation remarks. The innovative fabrication of NCs from CT/FM/SeNPs is strongly suggested to protect citrus crops from green mold and preserve fruits quality.

Fabrication of bioactive nanocomposites from chitosan, cress mucilage, and selenium nanoparticles with powerful antibacterial and anticancerous actions.

Natural bioactive alternatives are the utmost requests from researchers to provide biosafe and effectual health-guarding agents. The biopolymers chitosan nanoparticles (NCT), mucilage of cress seed (GCm; Lepidium sativum), and GCm-mediated selenium nanoparticles (GCm/SeNPs) were innovatively employed for fabricating novel bioactive natural nanocomposites (NCs) with elevated bioactivities as bactericidal (against Salmonella typhimurium and Staphylococcus aureus) and anticancer (against CaCo-2 and HeLa cells). The SeNPs were successfully generated with GCm, and different NCs formulations were fabricated from NCT:GCm/SeNPs amalgam ratios including T1, T2, and T3 with 2:1, 1:1, and 1:2 ratios, respectively. The infrared analysis of synthesized molecules appointed apparent physical interactions among interacted molecules. The average particles' sizes and charges of molecules/NCs were (12.7, 316.4, 252.8, and 127.3 nm) and (-6.9, +38.7, +26.2, and -25.8 mV) for SeNPs, T1, T2, and T3, respectively. The biocidal assessment of NCs indicated that T1 was the strongest antibacterial formulation, whereas T3 was the superior anticancer amalgam. These NCs formulations could exceed the biocidal potentialities of standard biocides. T1-NC could cause severe destructions/deformations in challenged S. typhimurium within 9 h, whereas T3-NCs induced apparent fluorescent apoptosis signs in treated HeLa cells. The prospective applications innovatively designed biocidal natural NCs that are recommended for controlling pathogenic bacteria and fighting cancerous cells.

Amelioration effect of 18ß-Glycyrrhetinic acid on methylation inhibitors in hepatocarcinogenesis -induced by diethylnitrosamine.

Aim: suppression of methylation inhibitors (epigenetic genes) in hepatocarcinogenesis induced by diethylnitrosamine using glycyrrhetinic acid. Method: In the current work, we investigated the effect of sole GA combined with different agents such as doxorubicin (DOX) or probiotic bacteria (Lactobacillus rhamanosus) against hepatocarcinogenesis induced by diethylnitrosamine to improve efficiency. The genomic DNA was isolated from rats' liver tissues to evaluate either methylation-sensitive or methylation-dependent resection enzymes. The methylation activity of the targeting genes DLC-1, TET-1, NF-kB, and STAT-3 was examined using specific primers and cleaved DNA products. Furthermore, flow cytometry was used to determine the protein expression profiles of DLC-1 and TET-1 in treated rats' liver tissue. Results: Our results demonstrated the activity of GA to reduce the methylation activity in TET-1 and DLC-1 by 33.6% and 78%, respectively. As compared with the positive control. Furthermore, the association of GA with DOX avoided the methylation activity by 88% and 91% for TET-1 and DLC-1, respectively, as compared with the positive control. Similarly, the combined use of GA with probiotics suppressed the methylation activity in the TET-1 and DLC-1 genes by 75% and 81% for TET-1 and DLC-1, respectively. Also, GA and its combination with bacteria attenuated the adverse effect in hepatocarcinogenesis rats by altering potential methylomic genes such as NF-kb and STAT3 genes by 76% and 83%, respectively. Conclusion: GA has an ameliorative effect against methylation inhibitors in hepatocellular carcinoma (HCC) by decreasing the methylation activity genes.

Green synthesis of chitosan nanoparticles, optimization, characterization and antibacterial efficacy against multi drug resistant biofilm-forming Acinetobacter baumannii.

Chitosan nanoparticles (CNPs) are promising versatile cationic polymeric nanoparticles, which have received growing interest over last few decades. The biocompatibility, biodegradability, environmental safety and non-toxicity of the chitosan nanoparticles makes it preferred for a wide range of biological applications including agriculture, medical and pharmaceutical fields. In this study, CNPs were biosynthesized by aqueous extract of Eucalyptus globulus Labill fresh leaves as bio-reductant. Box-Behnken design in 29 experimental runs was used for optimization of different factors affecting the production of CNPs. The maximum yield of CNPs was 9.91 mg/mL at pH of 4.5, chitosan concentration of 1%, incubation time of 60 min and temperature of 50 °C. The crystallinity, particle size and morphology of the biosynthesized CNPs were characterized. The CNPs possess a positively charged surface of 31.1 mV. The SEM images of the CNPs confirms the formation of spherical form with smooth surface. The TEM images show CNPs were spherical in shape and their size range was between 6.92 and 10.10 nm. X-ray diffraction indicates the high degree of CNPs crystallinity. FTIR analysis revealed various functional groups of organic compounds including NH, NH2, C-H, C-O, C-N, O-H, C-C, C-OH and C-O-C. The thermogravimetric analysis results revealed that CNPs are thermally stable. The antibacterial activity of CNPs was determined against pathogenic multidrug-resistant bacteria, Acinetobacter baumannii. The diameters of the inhibition zones were 12, 16 and 30 mm using the concentrations of 12.5, 25 and 50 mg/mL; respectively. When compared to previous studies, the biosynthesized CNPs produced using an aqueous extract of fresh Eucalyptus globulus Labill leaves have the smallest particle sizes (with a size range between 6.92 and 10.10 nm). Consequently, it is a promising candidate for a diverse range of medical applications and pharmaceutical industries.

Fabrication and assessment of potent anticancer nanoconjugates from chitosan nanoparticles, curcumin, and eugenol.

In cancer management and control, the most challenging difficulties are the complications resulting from customized therapies. The constitution of bioactive anticancer nanoconjugates from natural derivatives, e.g., chitosan (Ct), curcumin (Cur), and eugenol (Eug), was investigated for potential alternatives to cancer cells' treatment. Ct was extracted from Erugosquilla massavensis (mantis shrimp); then, Ct nanoparticles (NCt) was fabricated and loaded with Cur and/or Eug using crosslinking emulsion/ionic-gelation protocol and evaluated as anticancer composites against CaCo2 "colorectal adenocarcinoma" and MCF7 "breast adenocarcinoma" cells. Ct had 42.6 kDa molecular weight and 90.7% deacetylation percentage. The conjugation of fabricated molecules/composites and their interactions were validated via infrared analysis. The generated nanoparticles (NCt, NCt/Cur, NCt/Eug, and NCt/Cur/Eug composites) had mean particle size diameters of 268.5, 314.9, 296.4, and 364.7 nm, respectively; the entire nanoparticles carried positive charges nearby ≥30 mV. The scanning imaging of synthesized nanoconjugates (NCt/Cur, NCt/Eug, and NCt/Cur/Eug) emphasized their homogenous distributions and spherical shapes. The cytotoxic assessments of composited nanoconjugates using the MTT assay, toward CaCo2 and MCF7 cells, revealed elevated anti-proliferative and dose-dependent activities of all nanocomposites against treated cells. The combined nanocomposites (NCt/Eug/Cur) emphasized the highest activity against CaCo2 cells (IC50 = 11.13 µg/ml), followed by Cur/Eug then NCt/Cur. The exposure of CaCo2 cells to the nanocomposites exhibited serious DNA damages and fragmentation in exposed cancerous cells using the comet assay; the NCt/Eug/Cur nanocomposite was the most forceful with 9.54 nm tail length and 77.94 tail moment. The anticancer effectuality of innovatively combined NCt/Cur/Eug nanocomposites is greatly recommended for such biosafe, natural, biocompatible, and powerful anticancer materials, especially for combating colorectal adenocarcinoma cells, with elevated applicability, efficiency, and biosafety.

Semi-industrial Scale Production of a New Yeast with Probiotic Traits, Cryptococcus sp. YMHS, Isolated from the Red Sea.

A new yeast strain with promising probiotic traits was isolated from the Red Sea water samples. The isolate (YMHS) was subjected to genetic characterization and identified as Cryptococcus sp. Nucleotide sequence analysis of the rRNA gene internal transcribed spacer regions showed 95% sequence similarity between the isolate and Cryptococcus albidus. Cryptococcus sp. YMHS exhibited desirable characteristics of probiotic microorganisms; it has tolerance to low pH in simulated gastric juice, resistance to bile salts, hydrophobic characteristics, broad antimicrobial activity, and in vitro ability to degrade cholesterol. The isolate grew well in a semi-defined medium composed of yeast extract, glucose, KH2PO4, (NH4)2SO4, and MgSO4, yielding cell mass of 2.32 and 5.82 g/l in shake flask and in bioreactor cultures, respectively. Fed-batch cultivation, with controlled pH, increased the biomass gradually in culture, reaching 28.5 g/l after 32 h cultivation. Beside the feasible use as a probiotic, the new strain also could be beneficial in the development of functional foods or novel food preservatives. To our knowledge, this is the first report of yeast with probiotic properties isolated from the Red Sea.

Design, synthesis and QSAR study of certain isatin-pyridine hybrids as potential anti-proliferative agents.

A hybrid pharmacophore approach was adopted to design and synthesize new series of isatin-pyridine hybrids. All the newly prepared hybrids (5a-o, 8 and 11a-d) were in vitro evaluated for their anti-proliferative activity against three human cancer cell lines, namely HepG2 hepatocellular carcinoma, A549 lung cancer and MCF-7 breast cancer. Compound 8 emerged as the most active member against HepG2 cell line (IC50 = 2.5 ± 0.39 µM), with 2.7-fold increased activity than the reference drug, doxorubicin (IC50 = 6.9 ± 2.05 µM). Whilst, compound 11c was found to be the most potent counterpart against A549 and MCF-7 cell lines with IC50 values of 10.8 ± 1.15 and 6.3 ± 0.79, respectively. The weightiness of the utilization of non-cleavable linker, as the chalcone linker, and simplification of the first group, was explored via the SAR study. Furthermore, a QSAR model was built to explore the structural requirements controlling the cytotoxic activities. Notably, the predicted activities by the QSAR model were very close to those experimentally observed, hinting that this model could be safely applied for prediction of more efficacious hits comprising the same skeletal framework. Finally, a theoretical kinetic study was established to predict the ADME of the active hybrids.

Evaluation of newly isolated probiotics in the protection against experimental intestinal trichinellosis.

The potential use of probiotics in controlling enteric infections has generated tremendous interest in the last decade. The protective efficacy of seven oral doses of two newly isolated Egyptian probiotic strains; Lactobacillus acidophilus P110 (L. acidophilus) and Lactobacillus plantarum P164 (L. plantarum) versus Lactobacillus casei ATCC 7469 (L. casei) - against experimental intestinal trichinellosis - was assessed via parasitological, immunological and histopathological parameters, after verifying their in vivo safety and intestinal colonization. Parasitologically, the highest adult count reduction was observed in L. plantarum-fed infected sub-subgroup (56.98, 65.42 and 69.02%) - on the 5th, 12th and 17th days post infection (P.I.), respectively. Lesser percentage reductions were recorded in both the L. casei-fed infected sub-subgroup (36.19, 23.68 and 31.58%) and L. acidophilus-fed infected sub-subgroup (36.50, 11.8 and 7.61%) at the same intervals. On the 28th day post challenge, the highest larval count reduction was in L. plantarum-fed infected sub-subgroup (87.92%). While lower percentage yet still significant were observed in the L. casei-fed infected (74.88%) and L. acidophilus-fed infected sub-subgroups (60.98%). Immunologically, serum IFN-γ levels in the probiotic-fed non infected sub-subgroups were higher than those in the probiotic-fed infected sub-subgroups. Both showed higher levels of IFN-γ than the non probiotic-fed sub-subgroups. Histopathologically, intestinal sections of the probiotic-fed infected sub-subgroups showed amelioration of the inflammation and damage resulting from Trichinella spiralis (T. spiralis) infection. Results indicate that, through mechanical and immunological mechanisms, L. plantarum showed parasitological and histopathological protective superiority with respect to both L. casei and L. acidophilus against murine T. spiralis infection.

Analogue-based design, synthesis and biological evaluation of 3-substituted-(methylenehydrazono)indolin-2-ones as anticancer agents.

The docking studies on CDK2 and GSK-3ß inspired us to synthesis a series of indoline-2,3-dione hydrazones 10a-l. Treatment of indoline-2,3-dione derivatives 7a-d with hydrazine gave 3-hydrazonoindolin-2-ones 8a-d which were reacted with the appropriate aldehydes 9a-c to yield 3-substituted-(methylenehydrazono)indolin-2-ones 10a-l. Compounds 10a-l showed a significant anticancer activity against human breast cell line MCF-7. Compounds 10c, f, i exhibited the highest activity almost the same of doxorubicin (IC50 = 6.10 µM) with IC50 = 7.75, 6.75, 6.25 µM, respectively.