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This study evaluated the efficacy of entomopathogenic fungi (EPF) as biocontrol agents against aphids, whiteflies and western flower thrips. The research employed a leaf disc bioassay with various conidia concentrations to determine lethal concentration (LC) and time (LT) for pest eradication. Additionally, the study assessed the activity of cuticle-degrading enzymes produced by EPF (Chitinase, Protease, and Lipase) to understand their pathogenic mechanisms. Molecular identification using ITS region of 18S rDNA identified virulent isolates. Results indicated that four isolates, ENPF-16, 24, 41, and 60, achieved significant mortality rates (95% to 100%) at a concentration of 1x10⁸ conidia/mL after nine days. Akanthomyces sp. (ENPF-41) exhibited the highest enzyme activity, followed by Beauveria sp. (ENPF-60). The virulent fungal isolates were identified as Beauveria bassiana and Akanthomyces lecanii. Among EPFs, Akanthomyces lecanii (MT997935) displayed greater virulence against all three test insects with lower LC₅₀ and LT₅₀ values compared to other EPFs. In summary, all fungal isolates induced mortality in the tested pests, but their effectiveness varied. Akanthomyces lecanii (MT997935) emerged as a promising biocontrol candidate due to its broad host range and strong virulence.
 

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Ganoderma lucidum is one of the best-known medicinal mushrooms in the world. It contains substantial amounts of intra- and extracellular secondary metabolites and polysaccharides each with its own specific medicinal and medical uses. The chitin-glucan complex (CGC) is considered one of the important polysaccharides of this fungus. Among the 10 various culture media that were studied, the one containing PDB at 24g/l, peptone at 1g/l, and with the dry weight of cells of 11.6 g/l, the produced CGC of 3.2g/l, and with 27.6 percent CGC in the dry weight of the cells was selected as the suitable culture medium. FTIR analysis was performed for characterization of the produced CGC and its antibacterial properties were studied. The obtained time profile for CGC growth and production was 20 days and, using the logistic growth model and the Lodding-Pipet equation, the calculated specific growth rate of Ganoderma lucidum (μm) and the volumetric productivity for the product were 2.85 g CGC L-1day-and 0.5274 day-1, respectively. The calculations indicated there were high degrees of conformance between the model and the laboratory data related to kinetic characteristics of cell growth (R2= 0.9679) and to CGC production (R2=0.9901). Therefore, the introduced kinetic model can serve as an effective guide to control the fermentation process in industrial production of the valuable CGC polymer.

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Essential oils of four aromatic plants, Artemisia monosperma Del., Callistemon viminals (Sol.ex Gaertn.) G. Don, Citrus aurantifolia (Christm.) Swingle and Cupressus macrocarpa Hartw. ex Gordon, were evaluated for their anti-nutritional, antifeedant, growth inhibitory and insecticidal activities against Sopdoptera littoralis (Boisduval) (Lepidoptera: Noctuidae). The essential oils of A. monosperma and C. aurantifolia caused the highest reduction in relative growth rate (RGR) at the tested concentrations (125, 250, 500, 1000 and 2000mg/l). The RGR values ranged between 8.63 and 3.05 mg/day for A. monosperma, and between 10.74 and 2.89 mg/day for C. aurantifolia compared with 14.89 mg/day for control after 72 h of treatment. In general, the results showed that the values of relative growth rate (RGR) decreased with increasing the concentration of the tested oils. In addition, the tested oils significantly reduced efficiency of conversion of ingested food (ECI) and efficiency of conversion of digested food (ECD) values, particularly at the higher concentrations of 500, 1000 and 2000mg/l. On the other hand, the tested oils showed antifeedant activity against the larvae of S. littoralis with A. monosperma and C. aurantifolia oils being more active than C. viminals and C. macrocarpa oils. The tested oils showed remarkable growth inhibition effect as the growth inhibition index values were increased from 37.63 to 79.80% for A. monosperma, from 21.69 to 52.12% for C. viminals, from 16.55 to 28.59% for C. aurantifolia and from 37.64 to 52.32% for C. macrocarpa when the concentration increased from 125 to 2000mg/l. Based on chitin formation ratio values, the tested essential oils induced reduction in chitin formation. A. monosperma and C. macrocarpa essential oils revealed the highest insecticidal activity on 4^th instar larvae of S. littoralis. Examination of reproductive tracts of adult females emerged from treated larvae indicated that the tested oils caused undifferentiated ovarioles.

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Chitin is one of the most abundant renewable polysaccharides in nature that is widely found in the shell of the crustacean, insect cuticle and cell walls of fungi. Due to the unique properties such as biocompatibility, biodegradation and noun toxicity is widely used in various industries. In this study, we used of Banana shrimp, Penaeus merguinsis wastes (particle size 8-10 mm) to extract chitin using microbial fermentation method by Pseudomonas aeruginosa. Demineralization and deproteinization was carried out by incubating shrimp waste inoculated with bacteria at different concentration glucose (0%, 10%, 15% and 20% w/v) and inoculum (10%, 15% and 20% v/v) for 4 day in a shaking incubator (100 rpm) at 30°C. The results showed a direct correlation between the concentration of these parameters and deproteinization and demineralization rate. When studying the effect of these parameters, 20% glucose and 20% of the inoculum was determined as the optimum value, which leads to the production of chitin with a removal of minerals (76%) and protein (86%). Therefore, the microbial fermentation, as an ecofriendly and positive method, can be used to produce a high- quality chitin.

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Chitin and chitosan are two very important biopolymer products that have so many usages in the high cost industries. Chitin Converts into chitosan via de-acetylation of chitin. It occurs by alkaline melting method in the absence of oxygen. Chemical structure change, severe environmental pollution and De-polymerization are of the major problems in producing high quality chitosan. In this study for conversion of chitin into chitosan fungus Aspergillus niger strains (ATHUM-10864), the generator of de-acetylases enzymes were used instead of chemicals. Chitosan quality was determined via elemental analysis infrared spectroscopy, X-ray tomography, molecular weight determination and estimation of crystallinity percent, color and molecular structure.The results showed 80±5% efficiency in the conversion of chitin into chitosan or de-acetylation degree of chitin. The gained chitosan contained of 44.4 % carbon, 8.9 % nitrogen, 2.7 % hydrogen and 39.5 % oxygen. The physical characteristics were as 94.5% Crystallinity and pale brown color. The chemical structure of per unit of chitosan was obtained as C6H12NO4. The results showed that replacing biological methods instead of chemicals was possible to access well quality products. It also eliminates the use of chemical materials such as concentrate sodium hydroxide that is damaging the environment.

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The chitinase producing ability of Pseudomonas fluorescens strains viz., PF1, PB2 and FP7 was evaluated in a culture medium with and without a chitin source. The addition of 1% (v/v) chitin in culture medium significantly increased the bacterial population and chitinase activity. Among three strains tested, FP7 responded well to the addition of chitin by producing 31.2% increased chitinase in culture. Western blot analysis with chitinase antibody detected six and five chitinase isoforms in culture inoculated with FP7 and PF1, respectively.

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The trend toward sustainable development of the environment and economy has led to a large-scale debate on the use of seafood wastes. In recent years shrimp has been a major part of the food industry. The accumulated waste of shrimp without proper use has resulted in the destruction of the resources and problems of waste disposal and environmental pollution. Shrimp waste fermentation with microorganisms is a method for recovering biologically active material. Bacterial chitinase is considered as a degenerate enzyme .In this study, chitin degrading bacteria were isolated from different environment and then the most efficient strain was selected. The isolate identified by Microscopic, physiological and molecular characteristics and sequencing the 16SrRNA gene and compared with the Bacillus licheniformis strain, the highest rate of chitinase has been reported so far. The isolated strain identified as Bacillus altitudinis can ferment shrimp shell as the only sources of energy and produce high-temperature chitinase, with a 5.1 U/mL activity of over a period of 4 days, and 65.6 mg/l protein on semisolid shrimp shell. While it does not grow on the agar under normal conditions, therefore, its use can't cause pollution to the environment. As a result, the activity of chitinase, its simple and inexpensive method of concentration by heat, high enzyme resistance at high temperatures, activity in a wide range of pH and the use of cheap shrimp shell substrate show the superior functional quality of this strain in shrimp shell fermentation.
 

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Chitinases are essential enzymes in crustaceans that play an important role in the molting cycle and digestion of chitin. Based on the present study, the chitinase encoding cDNA of Penaeus mergueinsis with a length of 1440 bp containing 467 amino acids was sequenced by PCR and then its phylogenetic and bioinformatics analysis was performed. The new sequence was registered in the gene bank with the accessition number MT250539 and the molecular weight of the protein resulting from this sequence was predicted to be 51.84 KDa and the theoretical isoelectric point of 4.79. Comparison of amino acid sequences among penaeid chitinases showed the highest identification (about 97 to 92%) with P. mondon chi-3, F. chinensis, P. vannamei and P. japonicus chi-3, respectively. Phylogenetic studies showed that chitinase in the present study belongs to group 3 chitinases. Revealed protein pattern analyzes showed that chitinase from P. mergueinsis contained the catalytic domain Glyco-18 at position 2-347, a chitin-binding site of pritrophin A at position 403-456, a disulfide bridge formed by two cysteines at position 436-421 is a chitin-binding domain type 2, active site (¹¹7FDGLDMDWE¹²⁵), a proline / threonine-rich region at positions 376-412, and a putative N-glycosylation site at position 427-424 (NTSG). The present study shows that the P. mergueinsis sequence contains active chitinase motifs similar to previously sequenced chitinases, and in the case of cloning, expression and purification probably has functional and structural features similar to the enzymes of the above species.

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Aims: The purpose of the present study was to investigate the antioxidant and anti-inflammatory properties of glucosamine hydrochloride (G-HCl), glucosamine sulfate sodium chloride (GS-Na) and glucosamine sulfate potassium chloride (GS-K) isolated from the shells of Litopenaeus vannamei obtained from a shrimp processing plant.
Materials &Methods: G-HCl was synthesized via hydrolysis of chitin with concentrated HCl followed by several sequential decolorization, crystallization and washing steps. Using G-HCl as the precursor, addition of sodium and potassium sulfates at 40 ºC for 1 h resulted in production of GS-Na and GS-K.
Findings: The yield of chitin was found 19.9% and those of glucosamine products ranged between 75.5%-82.5%. The HPAEC-PAD indicated the presence of glucosamine monomers, as compared with commercial standard, with different elution time to that of glucose. The appearance of characteristic signals of O-H, N-H and C-O-C in the FT-IR spectra provided further support of glucosamine successful isolation. SEM images and EDX spectra of glucosamines confirmed the elemental compositions of samples and their polyhedral crystalline structures. DSC and TGA thermograms indicated endothermic and exothermic peaks specific to glucosamine products. Relatively low DPPH and ABTS radical scavenging activities and ferric reducing power was obtained for all glucosamine products. all the glucosamine derivatives indicated an anti-inflammatory effect on LPS-simulated RAW264.7 cells.
Conclusion: Glucosamine products showed no cytotoxicity and down-regulated the release of NO in RAW264.7 murine macrophage cells induced by LPS. Overall, the present results indicated the successful production of glucosamine from the waste of L. vannamei processing plant with antioxidant and anti-inflammatory properties.
 

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This study aimed to produce and investigate an emulsion film of gelatin by emulsion pickering method containing chitin nanoparticles. Different concentrations of nanochitin (0.2, 0.5, 1, and 2 g/g dry gelatin) and two different concentrations of corn oil (20% and 30% based on dry matter weight) were used to prepare the gelatin emulsion film by the emulsion pickering method. Then the properties of the films were investigated by examining the zeta potential, thickness, moisture and solubility, water vapor permeability (WVP), surface hydrophobicity, mechanical and thermal properties. The results showed that the gelatin emulsion containing 0.5 g/g of chitin nanoparticles was more stable than other samples. Nanochitin-stabilized emulsion films reduced moisture, solubility, and water vapor permeability compared to control films and tween-containing films, and nanochitin-containing films improved mechanical properties. In addition, the addition of nanochitin up to 0.5 g/g to the gelatin emulsion film improved the thermal properties because it led to an increase in melting temperature and enthalpy. The addition of nanochitin also increased the hydrophobic properties of the film. 30% oil concentration had a better effect on film properties than 20% oil. Therefore, the use of nanochitin as a stabilizing emulsion for pickering in gelatin films with a concentration of 30% corn oil can lead to the formation of biodegradable polymers with more acceptable properties for food packaging.

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Trichoderma species are known as effective agents used for biological control of plant pathogenic fungi. The Trichoderma harzianum and its mutant isolates were cultured and their traits including, mycelial growth, antagonistic activity and extracellular proteins and enzymes production (Chitinase and Cellulase) were investigated to select the most effective mutant isolates against plant pathogenic fungus Rhizoctonia solani. Also, the purity and composition of enzyme-rich protein samples were evaluated under denaturing gel electrophoresis. This study clearly showed the possibility of improving mycelia growth rate (from 1.18 to 1.33 cm d^-1), the antagonistic capability of Trichoderma (from 54.9% growth inhibition of R. solani to 66%), extracellular proteins and enzymes production for biological control of plant diseases through mutation with γ-radiation. Also, compared to wild type strain, protein production in the mutant isolates increased. Moreover, the highest specific chitinase enzyme activities were observed in mutant isolates T. h M8 (42.48 U mg^-1) and T. h M15 (38.25 U mg^-1). Trichoderma mutant of T. h M8 maintained higher mycelia growth rate and higher ability to inhibit growth of R. solani. The SDS-PAGE profiles had several enzyme protein bands such as CelloBioHydrolases (CBHs), EndoGlucanases (EGs), β-Glucosidases (BGLs), endochitinases, and β-(1, 4)-N-acetyl glucoaminidases. SDS-PAGE analysis indicated the presence of different protein bands in the range of 10.5 to 245 KDa. Interestingly, expression of chitinase in 95 percent of mutants was higher than wild type of T. harzianum. The results showed that gamma mutation could increase the efficiency and amount of enzymes in T. harzianum, while these enzymes are involved in antagonistic properties of T. harzianum.