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The current study evaluated the interactive impacts of water temperature and feeding rate on digestive enzymes, intestine histology, growth and stress-related genes, and cultivable intestinal microbiota of Asian seabass (Lates calcarifer). For this purpose, 180 fish (85.0±3.0 g) were reared at three different temperatures (20, 27, and 33 °C) and two feeding rates (apparent satiation and 2.5% of biomass) with three replications for 6 weeks. The results revealed no significant differences among different treatments regarding the activity of digestive enzymes (P˃0.05) of fish reared under different temperatures and feeding rates. The length, width, and thickness of intestinal villi were unaffected by different temperatures and feeding rates (P˃0.05). In addition, no variations were found in the total aerobic bacterial count of fish gut from different experimental groups (P˃0.05). At the molecular level, IGF-I and HSP70 coding genes were found to be highly expressed in experimental treatments (P<0.05). To conclude, present results showed that temperatures between 27 to 33 °C are more optimal for Asian seabass, and the different temperatures and feeding rates do not affect digestive enzymes, intestine histology, and gut microbiota after 6 weeks. Further molecular research is needed to unravel the complex impact and mechanisms of feeding rate and different rearing temperature on fish physiology.

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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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Biochemical characteristics of α-amylase in the digestive system and head glands of Cryptolaemus montrouzieri, a key predator of citrus mealybug, Planococcus citri (Pseudococcidae), were studied. The major isoform of α-amylase with the same molecular weight was detected in both gut and head glands loaded on polyacrylamide-starch gel electrophoresis. Moreover, a minor band with much lower intensity was observed in zymogram analysis of gut. Results showed that the specific activity of α-amylase from head glands (0.89 ± 0.02 μmol/min/mg protein) was significantly more than that of digestive system (0.76 ± 0.01 μmol/min/mg protein) in common condition (temperature equal to 25 ± 1 ^oC). The optimal pH and temperature for α-amylases activity were determined to be nearly 4 and 50°C in digestive system and 6 and 60 °C in head glands, respectively. EDTA (Ethylenediamine tetra acetic acid), Mg²⁺, Na⁺, Co²⁺, Fe²⁺ and Ca²⁺ inhibited the enzyme activity but Ba²⁺, Zn²⁺, Hg⁺ and K⁺ enhanced enzyme activity in digestive system. EDTA and all tested metal ions except Ba²⁺ inhibited the enzyme activity of head glands. Detectable levels of α-amylase activity in the insect reflect adaptation of the coccinellid for using starch granules or sugars (honeydew; sugary excreta of homopterans; and nectar) as a source of food in addition to predatory habits.

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Background: L-lysine is essential amino acid for human and animal nutrition. L-lysine is useful as medicament, chemical agent, food material (food industry) and feed additive (animal food). The industrial production of lysine has become an economically important industrial process. Several hundred thousands tones of L-lysine are produced annually worldwide, almost exclusively using Corynebacterium glutamicum. Study methods: To amplify LysA gene from C.glutamicum, two primers with NheI and HindIII restriction sites were designed. PCR was performed and PCR product was ligated with pTZ57R/T. Recombinant plasmid sequence was determined. LysA with sticky end was ligated with digested pET28a vector and ligation mixture was transformed in E.coli BL21(DE3). The recombinant plasmid was isolated with enzymatic digestion and sequencing. Results: LysA gene, a fragment with 1.3 kb, was cloned. PCR products and enzymatic digestion of extracted vectors with HindIII and NheI, sequencing and SDS-PAGE confirmed the authenticity of cloning. Recombinant bacterial colonies were investigated and confirmed by two methods (PCR and enzymatic digestion). Conclusion: In this study for the first time, the expression rate of Meso- diaminopimelate decarboxylase enzyme (EC 4.1.1.20) in this expression vector was investigated and was increased significantly.

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The gill Structure and localization of Na⁺, K⁺-ATPase were examined through branchial arches of the Persian sturgeon, Acipenser persicus larvae (25 days-post-hatched, 417.3 mg). Studies were conducted through light microscopy (H&E Staining) and immunofluorescence for Na⁺, K⁺-ATPase. Results showed each gill consisted of four complete holobranches and opercular hemibranch. Each filament carried rows of lamellae consisting of a network of interconnecting blood lacunae, which were lined by pillar cell flanges. Pavement cells covered the outermost layer of the lamella and blood cells were found in lacunae. High density of ionocytes (529.73 per mm² of the gill tissue) was found at the base of the lamella, in the interlamellar regions, on the filaments and the septums. Ionocytes possessed large size and round basal nuclei. Ionocytes possessed strong immunofleurescence in their cytoplasm, especially in the basolateral sides because of high concentration of the enzyme. The results showed that the main structures of the gill has already been formed at this developmental stage of the Persian sturgeon, and along with its respiratory and excretory roles, it also plays an important role in osmoregulation.

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Brassica napus is an important oilseed crop and the yield loss due to fungal disease stem rot caused by Sclerotinia sclerotiorum is a serious problem in cultivation of this crop. The pathogenesis-related (PR) protein, glucanase, hydrolyzes a major cell wall component, glucan, of the pathogenic fungi and acts as a plant defense barrier. In this study, a β-1,3-glucanase (bgn13.1) gene was isolated from the biocontrol fungus Trichoderma virens-10 (showing the high β-glucanase activity) and cloned in pUC19 cloning vector. The cloned fragment was confirmed by molecular analysis and showed to contain two short introns, 52 and 57 bp and an open reading frame coding 761 amino acids. The bgn13.1 gene was over-expressed under the CaMV35S promoter in B. napus, R line Hyola 308. Transformation of cotyledonary petioles was achieved by pBIKH1 containing bgn13.1 gene via Agrobacterium tumefaciens LBA4404. The insertion of transgene was verified by the polymerase chain reaction (PCR) and genomic DNA Southern dot blotting in T0 generation. RT-PCR analysis indicated that the transgenic canola plants were able to transcribe the β-1,3 glucanase gene. Also, we used transgenic over-expression approach in order to investigate antifungal activity of expressed Bgn13.1 on S. sclerotiorum. The heterologous expressed Bgn13.1 of line # 7 and line # 10 compared with other lines showed stronger inhibition against hyphal growth of S. sclerotiorum with

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The probiotic effects of Saccharomyces cerevisiae and Aspergillus niger on the feeding efficiency, body composition, ammonia excretion, blood serum enzymes and the intestinal microbiota of juvenile beluga, Huso huso was investigated. The fish (31.8±2.81g) were randomly allocated into 12 oval tanks at a density of 30 individuals per tank with three replicates for each treatment,. The fish were fed either a basal diet (as control) or the diet supplemented with S. cerevisiae and A. niger (2×10⁶, 4×10⁶ and 6×10⁶ cells g^-1) for 8 weeks. The results indicated that the probiotic supplemented diet at 6×10⁶ (cells g^-1) significantly improved FCR and other nutritional indicators compared to the control treatment (p<0.05). Significant improvements (p<0.05) were also observed in ammonia excretion and blood serum enzymes between treatments. Total viable fungus and Lactobacillus spp. count were significantly improved in treatment compared to control (p<0.05). These results indicated that S. cerevisiae and A. niger improved feeding performance and blood serum enzymes of beluga.

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The cotton bollworm, Helicoverpa armigera (Hübner), is a serious pest on a wide range of economic crops in many parts of the world. In this study, digestive proteolytic and amylolytic activities of H. armigera larvae (3^rd to 5^th instars) were evaluated on five main host plants including chickpea (cv Hashem), cowpea (cv Mashhad), soybean (cv 033), navybean (cv Dehghan), and corn (cv SC 704) at 25 ± 1 °C, relative humidity of 65 ± 5% and a photoperiod of 16: 8 (L: D) hours. The results indicated that the highest enzyme activity was in 5^th instar. The highest general protease activity of 3^rd, 4^th and 5^thinstar larvae was found on corn. The larvae fed on corn had the lowest trypsin activity. This leads to hyperproduction of chymotrypsin and elastase-like enzymes to compensate the inhibition of trypsin. The larvae of H. armigera reared on cowpea had the highest level of amylase activity, and the lowest activity was in the larvae fed on corn. The results revealed that among host plants tested, corn was more resistant (unsuitable) to H. armigera. Study on digestive proteolytic and amylolytic activities of H. armigera can be used in identifying the antidigestive or antifeedent compounds, which will help us to design appropriate management programs.

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The aim of the present study was to evaluate the effects of enriched Artemia with fish and soybean oils supplemented with vitamin E on growth performance, stress resistance, antioxidant enzymes activity and lipid peroxidation of Persian sturgeon (Acipenser persicus) larvae. Five experimental diets including non-enriched  Artemia (control diet), Artemia enriched with soybean oil supplemented with 15 or 30% vitamin E (S15 and S30 diets) and fish oil supplemented with 15 or 30% vitamin E (F15 and F30 diets) were used. The larvae were fed to apparent satiation for 17 days. The results indicated that fish fed enriched Artemia had no significant differences compared with control group in terms of growth and survival, but increase in vitamin E levels from 15 to 30 % improved growth performance and resistance to salinity stress. Vitamin E content in fish fed S15 and S30 diets was significantly higher compared with the other treatments. Antioxidant enzymes activity in fish fed non-enriched Artemia, F15 and F30 diets were higher. The highest TBA value was observed in fish fed non-enriched Artemia. The results demonstrated that the addition of vitamin E to the fish and soybean oils for Artemia enrichment could reduce oxidation of oils and beneficial for the health and quality of larvae. In conclusion, enrichment of Artemia with soybean oil supplemented with 30 % vitamin E (S30 diet) is recommended for feeding Persian sturgeon larvae.

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Papain (EC2.22.4.3) is a thiol protease with high level of activity that has widespread industrial applications. The use of immobilized papain provides many advantages over its free form. In many applications, cysteine must be added as an activator. On the other hand, certain bivalent metal ions including Ca²⁺ behave as the inhibitors of papaein. In the present study, after preparation of Sepharose 6B with CNBr, a 5 mg/ml-protein solution was added to activate the gel for covalent attachment of enzyme and, subsequently, 2M glycine solution was added to block the remaining active groups on the gel.  The immobilization process brought about significant enhancement of storage, thermal stability, stability at extreme pHs, and resistance against the inhibitory effect of bivalent metal ions with respect to papain. The optimum temperature of papain was increased by 20 °C (from 60 to 80 °C) and its optimum pH was shifted from 7 to 8.0 upon immobilization. Also k_m and k_cat of the enzyme altered due to the immobilization process.These results are important in particular if one considers that the major problem in enzyme immobilization is the loss of enzyme activity and catalytic efficiency.

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A 3*2*2 factorial experiment was conducted to study the effect of feeding whole wheat (0, 10, and 20%), a microbial enzyme source (Arabino-xylanase based enzyme for wheat at 0% and 0.1%) and 0% or 1% insoluble grit (number 3 layer size) on the performance of White Leghorn hens at 19 to 54 weeks of age. Each dietary treatment was replicated four times with ten hens, housed two to a cage, each. No main treatment effects or interactions were observed for egg production, feed efficiency, egg weight, body weight gain, or mortality. Feeding 20% whole wheat produced the lowest incidence of cracked, broken, or soft shelled eggs. Adding the enzyme significantly (P<0.05) reduced daily feed intake from 124.6 to 122.2 g. Enzyme addition also resulted in minor, but significant (P<0.05), increases in egg specific gravity from 1.0800 to 1.0806. Inclusion of 20% whole wheat does not adversely affect the productivity of White Leghorn hens and therefore can be used to reduce the cost of feed processing.

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Background: Parasitic infections can cause different chemical changes in human body. This study was designed to determine the liver aminotransferase changes in Iranian people infected by parasite.
Materials and Methods: In this cross-sectional study, blood samples were collected from 183 parasite-infected patients. After serum isolation in laboratory, Alanine aminotransferase (ALT) and Aspartate aminotransferase (AST) rates of all samples were measured according to IFCC standard method, and after recording in the information form, the results were analyzed using descriptive statistics.
Results: The results of this study showed that patients were infected by 19 different types of parasites. Also, the results showed that the average value ​​of ALT and AST in patients were equal to 32.30 ±23.40 and 38.60±34.80, respectively. The ANOVA test showed remarkable statistical differences between these values ​​and various infections. There was positive and significant correlation between ALT and AST.
Conclusion: The abnormality in liver function in patients with eosinophilia can be caused by parasitic infections in endemic areas; therefore, it can be the cause of a differential diagnosis for physicians.  Generally, non-significant changes in transaminase levels can also be attributed to the infection low intensity in patients tested.

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The use of enzymes in organic solvents has biotechnological and industrial importance. Organic solvents can decrease the stability of enzymes that is a challenge for the use of enzymes in organic media. There are several approaches such as protein engineering, chemical modification, and use of additives for stabilization of enzymes in organic solvents. In this study, activity and stability of trypsin were investigated in the presence of different organic solvents. Then the effect of sucrose on the stability of the enzyme was investigated in the absence and prescence of solvents. The result showed that the activity and stability of trypsin were decreased in the presence of organic solvents. DMF had a lowest effect on the activity and stability of the enzyme. The use of sucrose increased the stability of trypsin in the presence of organic solvents. The stabilization effect of sucrose in the presence of DMF was more than other solvents. Consequently, a mixture of DMF and sucrose is proposed for the use of trypsin in industrial applications.

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The production of bioethanol from lignocellulosic biomass could be considered as an appropriate and economic option to remove environmental disasters and improve energy security. In fact, lignocellulosic material is mainly composed of cellulose, hemicellulose and lignin. Lignin works as the adhering prevents the bioconversion of cellulose into sugars and ultimately to ethanol. To address the problem, various chemical, physical, physicochemical and biological methods have been suggested. Enjoying convenient operating conditions, production of non-hazardous wastes, and having no harmful side effects, make the biological methods a potentially proper option. Unfortunately, the biological methods are slower and less efficient in comparison with the other processes. In the present study, an attempt is made to resolve this problem in an enzymatic degradation of lignin of a rice straw sample. Several peroxidase enzymes were produced by a white rot fungus, and their effects on lignin removal from the biomass samples were investigated in shaking flasks. Lignin concentration and enzymes' activity were measured by the acetyl bromide-soluble lignin spectrophotometric method and optical density method using special reagents, respectively. The results revealed that the enzymatic treatment could remove at least 30% of the lignin content of the lignocellulosic biomass. To achieve the maximum activity of the enzymes, The chemical composition of the culturing medium was optimized for the concentration of important metal ions including Cu2+, Mn2+ and Zn2+ through Box Behnken response surface methodology. The enzymes' activity at the obtained optimal conditions increased four times for Manganese peroxidase, and lignin peroxidase.

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The effect of simultaneous application of probiotic and digestive enzyme on the intestinal digestive activity of the whiteleg shrimp, Litopenaeus vannamei, was evaluated. For this purpose, the juvenile shrimps were fed for 30 days with a dietary probio-enzyme (containing a combination of six exogenous enzymes and four probiotic bacterial strains) at four concentrations of 0, 0.25, 0.5, and 1 g kg^-1 feed. Shrimps (5.04±0.39 g ind^-1) were randomly distributed in 12 aquaria (4 treatments × 3 replications); each aquarium contained 15 individual shrimps. Results indicated significantly (p≤0.05) higher growth performance, amylase and lipase activity at 0.5 g kg^-1 treatment as compared to other treatments. Protease activity was, however, significantly (p≤ 0.05) higher at 1.0 g kg^-1treatment as compared to other groups. Results also indicated that increase in the concentration of probiotic and enzymes supplementation was not associated with increase in all the digestive enzyme activity. In other words, probiotics and enzymes onlywithin specific range can have positive effect on growth performance and digestive enzyme activity of L. vannamei, above or below.

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Effects of feeding Artemianauplii enriched with PHB (0, 0.1, 0.3 and 1 g/L concentration) on the growth performance, body composition, digestive enzyme activity and hindgut bacterial community in the Persian sturgeon hatchlings were investigated. PHB treatment significantly (p≤ 0.05) decreased growth performances of the hatchlings. The PHB also significantly increased the total saturated fatty acids (SFA) and n6, but decreased the total MUFAs, C18:3n3, n3 and n3/n6.  PHB also altered digestive enzyme by significantly decreasing  the total protease, amylase, and lipase. Based on molecular analysis, PHB changed the microbial community in the hindgut of the hatchlings where less dominant bands were observed. Our results show that PHB has negative effects on the Persian sturgeon hatchlings.  Further studies are needed to find out the optimal concentration of PHB to apply in early larval rearing of sturgeon.

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The effects of starvation periods on physiological response of the juvenile Persian sturgeon, Acipencer persicus, was assessed through such stress factors as glucose, cortisol and hepatic enzymes for a period of 8 weeks. For this purpose, in a randomly designed experiment, five groups of fish (108.04 ± 0.28 gr) in 3 replicates were starved for a period of 0 (control), 1, 2, 3 and 4 weeks and fed them to satiation after their starvation lags. Blood plasma glucose and cortisol during starvation periods did not significantly change (p>0.05). This indicates high performance of this specific in maintenance of blood glucose during starvation and recovery of glucose level after feeding. However, plasma hepatic enzymes level in fasting treatments increased (p<0.05), but reached the control level after 4 weeks of feeding. Considering the role of liver enzymes in gluconeogenesis and taking into consideration the constant plasma glucose and cortisol during periods of food deprivation, it seems that liver enzymes in the Persian sturgeon during food deprivation play a key role in stabilizing blood glucose. Considering the role of liver enzymes in gluconeogenesis, and taking into consideration the constant plasma glucose and cortisol during periods of food deprivation, In this study, it seems that liver enzymes in the Persian sturgeon during food deprivation play a key role in stabilizing blood glucose.

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Laccase enzymes are polyphenol oxidase that catalyze the oxidation of wide range of phenolic components including phenols, polyphenols, aromatic amines and non-phenolic substitution with molecular oxygen as electron acceptor. So these enzymes have biotechnological application such as wastewater treatment system, bioremediation of soil pollution and etc. Result from previous studies showed an increase in thermal stability of bacterial laccase from Bacillus sp. HR03 using site directed mutagenesis and the effect of E188 residue on the surface regions at the interface between domain 1 and 2 in stability was confirmed. The aim of the present work was to investigate the effect of this amino acid substitution on enzyme activity in the presence of dimethyl sulfoxide and dimethylformamide as organic solvents. Compression of kinetic parameters including Kcat / Km ، ∆∆G‡, C50 showed significant increases in the mutant enzyme than wild type enzyme, that industrial application of the enzyme will be easy.