Levels Of Cellulase Research Articles

Alteration of Cecal Microbiota by Antimicrobial Peptides Enhances the Rational and Efficient Utilization of Nutrients in Holstein Bulls.

We previously observed that supplementation with antimicrobial peptides facilitated the average daily weight gain, net meat, and carcass weights of Holstein bulls. To expand our knowledge of the possible impact of antimicrobial peptides on cecum microbiota, further investigations were conducted. In this study, 18 castrated Holstein bulls with insignificant weight differences and 10 months of age were split randomly into two groups. The control group (CK) was fed a basic diet, whereas the antimicrobial peptide group (AP) was supplemented with 8 g of antimicrobial peptides for 270 days. After slaughter, metagenomic and metabolomic sequencing analyses were performed on the cecum contents. The results showed significantly higher levels of amylase, cellulase, protease, and lipase in the CK than in the AP group (P ≤ 0.05). The levels of β-glucosidase and xylanase (P ≤ 0.05), and acetic and propionic acids (P ≤ 0.01), were considerably elevated in the AP than in the CK group. The metagenome showed variations between the two groups only at the bacterial level, and 3258 bacteria with differences were annotated. A total of 138 differential abundant genes (P < 0.05) were identified in the CAZyme map, with 65 genes more abundant in the cecum of the AP group and 48 genes more abundant in the cecum of the CK group. Metabolomic analysis identified 68 differentially expressed metabolites. Conjoint analysis of microorganisms and metabolites revealed that Lactobacillus had the greatest impact on metabolites in the AP group and Brumimicrobium in the CK group. The advantageous strains of the AP group Firmicutes bacterium CAG:110 exhibited a strong symbiotic relationship with urodeoxycholic acid and hyodeoxycholic acid. This study identified the classification characteristics, functions, metabolites, and interactions of cecal microbiota with metabolites that contribute to host growth performance. Antimicrobial peptides affect the cecal microorganisms, making the use of nutrients more efficient. The utilization of hemicellulose in the cecum of ruminants may contribute more than cellulose to their production performance.

Functional analysis of microorganisms and metabolites in the cecum of different sheep populations and their effects on production traits.

The purpose of this study was to investigate the effects of intestinal microbiota on the growth and production performance of different groups of sheep, focusing on the role of cecal microbiota in regulating intestinal function, enhancing digestion and absorption, and improving feed utilization. The production performance of MG × STH (Mongolia × Small Tailed Han) F1 hybrids and purebred STH (Small Tailed Han) sheep by measuring various factors, including enzyme activities and VFAs (volatile fatty acids), to analyze changes in cecal fermentation parameters across different sheep groups. Metagenomic and metabolomic sequencing combined with bioinformatics to analyze the cecal contents of the two sheep populations. The study findings indicated that the MG × STH F1 hybrids outperformed the purebred STH in terms of body weight, height, oblique body length, and VFAs (p < 0.05). Additionally, the MG × STH F1 higher levels of protease and cellulase in the cecum compared to the purebred sheep (p < 0.05). Metagenomic analysis identified 4,034 different microorganisms at the species level. Five differential organisms (Akkermansiaceae bacterium, Escherichia coli, unclassified p Firmicutes, Streptococcus equinus, Methanobrevibacter millerae) positively regulated sheep performance. Metabolomics identified 822 differential metabolites indoleacetaldehyde, 2-aminobenzoic acid, phenyl-Alanine, enol-phenylpyruvate and n-acetylserotonin were associated with improved performance of sheep. The combined results from the metagenomic and metabolomic studies suggest a positive correlation between specific microbes and metabolites and the performance of the sheep. In conclusion, the MG × STH F1 hybrids demonstrated superior growth performance compared to the purebred STH sheep. The identified microorganisms and metabolites have promising roles in positively regulating sheep growth and can be considered key targets for enhancing sheep performance.

Evaluation of Himalayan nettle (Urtica ardens) extract meal in common carp (Cyprinus carpio) feed: Impact on growth performance, digestive enzyme activity and antioxidant capacity

This research aims to evaluate the impact of an extract derived from the leaves of Himalayan nettle (Urtica ardens) leaf extract on common carp, Cyprinus carpio, growth, digestive enzymes, fish body carcass composition, antioxidant system, and water quality parameters. For a duration of 90 days, experimental diets containing nettle extract were given to 150 fish (12.38 ± 0.53 g) in 15 tanks (10 fish per tank). The five groups in triplicate were: control or T0 (untreated), T1 (fed 0.3% extract), T2 (0.6% extract), T3 (0.9% extract), and T4 (1.2% extract). Results revealed that nettle extract in the diet increases common carp growth up to 0.9% concentration of extract. Its feed conversion ratio decreases, while weight gain and specific growth rate rise significantly compared to the control diet. No mortality was observed during the 90-day experiment. Dietary nettle extract did not affect whole-body fibre and crude ash percentages (p > 0.05). Nettle-fed and control groups showed significant differences in crude protein, body fat, and total body moisture (p < 0.05). After a 90-day feeding trial, 0.9% nettle extract-fed fish had higher protease, amylase, lipase, and cellulase levels than the control group (p < 0.05). When nettle content reached 1.2%, enzyme activity decreased. Orthophosphate production peaked at 4 and 8 hours after feeding, while ammonia excretion peaked at 2 and 6 hours. The control group excreted the most ammonia and orthophosphate, while the 0.9% group excreted the least. Liver and kidney SOD levels were higher in common carp fed a 0.6% nettle extract diet. Feeding with a 0.9% nettle extract diet improved hepatic lipid peroxidation, gut and liver glutathione. Gut and kidney SOD, catalase, LPO, and GSH increased dramatically on the 1.2% diet. Statistically, the 0.9% nettle extract (T3) group showed the overall best results compared to the control diet. These findings demonstrate the potential of nettle extract as a dietary supplement in aquaculture.

INFLUENCE OF CULTIVATION CONDITIONS ON CELLULASE AND XYLANASE ACTIVITY OF RHIZOPUS ORYZAE F-1030

The paper examines the relationship between the activity of cellulase and xylanase enzymes and the lactic acid yield which was synthesized by the fungus Rhizopus oryzae F-1030 during its cultivation on a culture medium based on catalytically modified neutral sulfite liquors. It was shown that in cultivating by the deep method the fungus R. oryzae F-1030 synthesizes xylanase and cellulase enzymes that break down polysaccharides in a nutrient medium to simple sugars which are easily accessible for assimilation by the fungus. Accordingly, the fungus R. oryzae F-1030 can be used as a promising biotechnological object for the bioconversion of cellulose production secondary resources. It was found that the level of cellulase and xylanase expression in R. oryzae F-1030 depends on the type of substrate. The xylanase activity of the fungus is more pronounced than the cellulase activity since xylan oligomers predominate in birch wood. It was noted that the lactic acid yield during the cultivation of the fungus R. oryzae F-1030 on a catalytically modified neutral sulfite liquor depends on the action specifics of the hydrolyzing catalyst. As a nutrient medium for the microbiological synthesis of lactic acid the fungus R. oryzae F-1030 can use biocatalytically treated liquors of birch wood neutral sulfite pulping.

Sweet potato (Ipomoea Batatas (L.) Lam.): A study on physiological and biochemical properties

This study investigates the physiological and biochemical properties of different sweet potato (Ipomoea batatas (L.) Lam.) varieties grown in the Samarkand soil-climatic conditions and assesses the impact of these varieties on soil enzyme activities. The red, yellow, and white varieties were evaluated for their carbohydrate content, vitamin levels, antioxidant activities, growth parameters, root system development, and enzyme activities, including protease, amylase, and cellulase. Biochemical analysis revealed that the red variety had the highest carbohydrate content (25%) with significant levels of sucrose, glucose, and fructose. It also exhibited the highest vitamin C (30 mg/100g) and vitamin A (8 mg/100g) concentrations as determined by High-Performance Liquid Chromatography (HPLC). Antioxidant activity, measured using the DPPH assay, was highest in the red variety (85% scavenging activity). Physiological analysis showed that the red variety had superior growth parameters with a height of 35 cm, root length of 20 cm, and biomass of 150 g. It also demonstrated the most developed root system with a root weight of 50 g and root volume of 40 cm³. Enzyme activity assays indicated that the red variety had the highest levels of protease (85 U/mg), amylase (70 U/mg), and cellulase (65 U/mg). The study highlights the significant variations among sweet potato varieties in terms of their nutritional and health-promoting characteristics. The red variety emerged as the most beneficial, exhibiting superior nutritional content, antioxidant activity, and enzyme activities that enhance soil health. These findings underscore the potential of red sweet potatoes in promoting sustainable agriculture and improving food security in regions with challenging environmental conditions.

Molecular characterization of the ligninolytic Sundarban mangrove isolate Stenotrophomonas maltophilia GD1 and its promising applications in the dye and fiber industry

Lignocellulolytic biocatalysts, including laccase, cellulase, and pectinase, have high potential for degrading lignocellulosic biomass (LCB). Jute and banana are fiber-producing LCBs that hold significant economic value. Nevertheless, certain parts of these plants are utilized as waste material due to their low-quality fiber, which causes losses for the industry. These waste portions can be processed and biologically converted into high-value fiber. A bacterial strain called Stenotrophomonas maltophilia GD1, which can produce several proteins, was isolated from the Sundarban mangrove ecosystem in West Bengal, India, and genetically analyzed via 16S rRNA sequencing. The strain exhibited the ability to produce all three necessary lignocellulolytic enzymes, with the highest levels of laccase and minimal cellulase and pectinase activity. The strain modified the fibers extensively, as determined by Young’s modulus, and removed the color from the toxic industrial dyes. This demonstrates the broad potential of this strain for waste valorization and environmental remediation. Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) confirmed the enhanced fiber quality. Microbial phylogenetic relationships were established using Molecular Evolutionary Genetics Analysis version 11 (MEGA 11).

Enhanced rice plant (BRRI-28) growth at lower doses of urea caused by diazinon mineralizing endophytic bacterial consortia and explorations of relevant regulatory genes in a Klebsiella sp. strain HSTU-F2D4R

Endophytic biostimulant with pesticide bioremediation activities may reduce agrochemicals application in rice cultivation. The present study evaluates diazinon-degrading endophytic bacteria, isolated from rice plants grown in the fields with pesticide amalgamation, leading to increased productivity in high-yielding rice plants. These endophytes showed capabilities of decomposing diazinon, confirmed by FT-IR spectra analysis. Growth promoting activities of these endophytes can be attributed to their abilities to produce an increased level of IAA content and to demonstrate high level ACC-deaminase activities. Furthermore, these endophytes demonstrated enhanced level of extracellular cellulase, xylanase, amylase, protease and lignin degrading activities. Five genera including Enterobacter, Pantoea, Shigella, Acinetobacter, and Serratia, are represented only by the leaves, while four genera such as Enterobacter, Escherichia, Kosakonia, and Pseudomonas are represented only by the shoots. Five genera including, Klebsiella, Enterobacter, Pseudomonas, Burkholderia, and Bacillus are represented only by the roots of rice plants. All these strains demonstrated cell wall hydrolytic enzyme activities, except pectinase. All treatments, either individual strains or consortia of strains, enhanced rice plant growth at germination, seedling, vegetative and reproductive stages. Among four (I-IV) consortia, consortium-III generated the maximum rice yield under 70% lower doses of urea compared to that of control (treated with only fertilizer). The decoded genome of Klebsiella sp. HSTU-F2D4R revealed nif-cluster, chemotaxis, phosphates, biofilm formation, and organophosphorus insecticide-degrading genes. Sufficient insecticide-degrading proteins belonging to strain HSTU-F2D4R had interacted with diazinon, confirmed in molecular docking and formed potential catalytic triads, suggesting the strains have bioremediation potential with biofertilizer applications in rice cultivation.

Influence of light regulation on growth and enzyme production in rare endolichenic fungi.

The influence of light regulation on the growth and enzyme production of three endolichenic fungal isolates, i.e. Pseudopestalotiopsis theae (EF13), Fusarium solani (EF5), and Xylaria venustula (PH22), was determined. The isolates were exposed to blue, red, green, yellow, white fluorescent light (12h light-12h dark photoperiod) (test), and 24h dark (control) conditions. Results revealed that the alternating light-dark conditions resulted in the formation of dark rings in most fungal isolates but was absent in PH22. Red light induced sporulation while yellow light elicited higher biomass in all isolates (0.19 ± 0.01g, 0.07 ± 0.00g, and 0.11 ± 0.00g, for EF13, PH22, and EF5, respectively) as compared to incubation in the dark. Results also showed that blue light induced higher amylase activity in PH22 (15.31 ± 0.45 U/mL) and L-asparaginase activity in all isolates (0.45 ± 0.01 U/mL, 0.55 ± 0.39 U/mL, and 0.38 ± 0.01 U/mL, for EF13, PH22, and EF5, respectively) compared to both control conditions. Green light enhanced the production of xylanase (6.57 ± 0.42 U/mL, 10.64 ± 0.12 U/mL, and 7.55 ± 0.56 U/mL for EF13, PH22, and EF5, respectively) and cellulase (6.49 ± 0.48 U/mL, 9.57 ± 0.25 U/mL, and 7.28 ± 0.63 U/mL, for EF13, PH22, and EF5, respectively). In contrast, red light was the least effective light treatment as production of enzymes was the least, with lower levels of amylase, cellulase, xylanase, and L-asparaginase detected. To conclude, all three endolichenic fungi are light-responsive, with fungal growth regulated with the use of red light and yellow light, and manipulation of enzyme production via blue and green light.

SYNERGISTIC EFFECT OF HONEY AND LEMONJUICE-ENRICHED MULBERRY DIETS ON THE DIGESTIVE METABOLISM OF THE SILKWORM, BOMBYX MOR

Synergistic impact of honey and lemon juice-enriched mulberry diets has been studied on the digestive metabolism of fth instar larval Bombyx mori. The study focused on the digestibility of proteins, carbohydrates, sucrose and cellulose by their digestive enzymes in the larval midgut wall and midgut lumen compartments and biomass accumulation in the gut wall. The honey and lemon juice-enriched diets showed positive impact on protease activity and protein digestion, but did not show discernable effect on the activity levels of α-amylase, sucrase and cellulase and the digestibility of carbohydrates, sucrose and cellulose. Nevertheless, they signicantly reinforced the digestive mass accumulation in gut wall cells in accordance with the Hutchinson's investment principle. Further, as evidenced by higher growth rates in the digestive parameters during rst ve days of the fth instar regime, the rate of biomass accumulation is ne-tuned by the timing of acquisition of critical larval body size determinants.

Regulator of G Protein Signaling Proteins Control Growth, Development and Cellulase Production in Neurospora crassa.

Heterotrimeric (αβγ) G protein signaling pathways are critical environmental sensing systems found in eukaryotic cells. Exchange of GDP for GTP on the Gα subunit leads to its activation. In contrast, GTP hydrolysis on the Gα is accelerated by Regulator of G protein Signaling (RGS) proteins, resulting in a return to the GDP-bound, inactive state. Here, we analyzed growth, development and extracellular cellulase production in strains with knockout mutations in the seven identified RGS genes (rgs-1 to rgs-7) in the filamentous fungus, Neurospora crassa. We compared phenotypes to those of strains with either knockout mutations or expressing predicted constitutively activated, GTPase-deficient alleles for each of the three Gα subunit genes (gna-1Q204L, gna-2Q205L or gna-3Q208L). Our data revealed that six RGS mutants have taller aerial hyphae than wild type and all seven mutants exhibit reduced asexual sporulation, phenotypes shared with strains expressing the gna-1Q204L or gna-3Q208L allele. In contrast, Δrgs-1 and Δrgs-3 were the only RGS mutants with a slower growth rate phenotype, a defect in common with gna-1Q204L strains. With respect to female sexual development, Δrgs-1 possessed defects most similar to gna-3Q208L strains, while those of Δrgs-2 mutants resembled strains expressing the gna-1Q204L allele. Finally, we observed that four of the seven RGS mutants had significantly different extracellular cellulase levels relative to wild type. Of interest, the Δrgs-2 mutant had no detectable activity, similar to the gna-3Q208L strain. In contrast, the Δrgs-1 and Δrgs-4 mutants and gna-1Q204L and gna-2Q205L strains exhibited significantly higher cellulase activity than wild type. With the exception of sexual development, our results demonstrate the greatest number of genetic interactions between rgs-1 and gna-1 and rgs-2 and gna-3 in N. crassa.

170 Improving Nutrient Value of Soybean Meal Using Characterized Novel Microbial Fermentation

Abstract Soybean meal (SBM) is a common plant-based protein source in the swine industry. Its crude protein content is 44 to 50%. However, the presence of antinutritional factors such as high molecular weight, difficult to digest proteins, phytate and non-starch polysaccharides (NSP) limit its use in pigs. We aimed to use fermentation approach to improve the nutrient value of SBM. We have screened, from different sources, and identified bacterial strains that secrete cellulase, xylanase and proteinase as candidates for feedstuff fermentation. Further studies revealed that a few of them are capable of breakdown glycinin and beta-conglycinin, the major proteins in SBM that are difficult to digest and often resulted in intestinal upset in monogastric animal. Molecular characterization revealed that isolate 9 is a Bacillus subtilis strain (B. subtilis 9) secreted high level of cellulase, xylanase, and proteinase comparing to the ATCC control strain and other isolates. It grew significantly faster than the control strain. In addition, it also survived a wide range of concentrations salts and pH. When used for SBM fermentation, protein profiling of fermented SBM using SDS-PAGE revealed a significant reduction of high molecular weight proteins, especially those of glycinin and beta-conglycinin, to small peptides. On a dry matter basis, neutral detergent fiber (NDF) of the fermented SBM was decreased by 34%; crude protein content was increased from 49 to 57%. Pigs fed with a diet formulated with the fermented SBM as the sole source of crude protein had significantly higher apparent ileal digestibility (AID) of dry matter (80 vs. 72 %), ash (56 vs. 36%), crude protein (84 vs. 78%), NDF (71 vs. 66%), and acid detergent fiber (62 vs. 53%) compared with the control pigs fed with unfermented soybean meal. In addition, we have recently explored the possibility of improving feed stuff fermentation efficiency combining microbial species, the effect of this approach will be discussed.

Links between chemical composition of soil organic matter and soil enzyme activity in alpine grassland ecosystems of the Tibetan Plateau

Soil enzymes play a key role in soil organic matter dynamics in terrestrial ecosystems; however, the relationship between soil enzyme activity and the chemical composition of soil organic matter remains unclear. We analysed the soil organic matter (SOM) of five types of alpine grassland ecosystems in the Tibetan Plateau at the molecular level using pyrolysis gas chromatography/mass spectrometry (Py-GC/MS), which allowed us to examine the relationship between the patterns of enzyme activities and SOM composition. We found that the level of activity of carbon (C)-cycling enzymes (invertase, β-1,4-glucosidase, polyphenol oxidase, and peroxidase), nitrogen (N)-cycling enzymes (arylamidase), and phosphorus (P)-cycling enzyme (alkaline phosphatase) was significantly different among the five grassland ecosystems. In addition, we found significant variation among the different grassland ecosystems in terms of the activity of enzymes involved in C-cycling less so with those enzymes involved in N-cycling. In general, SOM was dominated by N-compounds (∼32.47–51.76%), polysaccharides (∼10.67–20.42%), and fatty acids (∼7.03–20.33%) in the different alpine grasslands. The most abundant compounds in SOM were D-alanine (10.06–20.77%), 9-octadecenamide, (Z) (4.43–9.68%), and 13-docosenamide (1.52–14.76%). Correlation analysis showed that the N-compound abundance was negatively correlated with nitrate reductase. Polysaccharides were positively correlated with activity levels of invertase and cellulase. Fatty acids were positively correlated with polyphenol oxidase activity and negatively correlated with alkaline phosphatase activity. Some abundant compounds consistently showed strong correlations with activity levels of invertase and cellulase, including toluene, styrene, benzene, phenol, furfural, pyrrole, p-cresol, 2-methylphenylacetylene and indole (positive correlation), 13-docosenamide, dodecanamide and o-hydroxybiphenyl (negative correlation). Furthermore, Gram-positive and Gram-negative bacteria also play a significant role in regulating SOM-enzyme interactions. These findings confirmed the strong effects of SOM composition on the activity of invertase and cellulase, indicating that invertase and cellulase play an important role in regulating SOM to some extent, resulting in complex interactions among organic-enzyme-microbes.

Effect of shiitake mushrooms polysaccharide and chitosan coating on softening and browning of shiitake mushrooms (Lentinus edodes) during postharvest storage

We investigated the browning and softening of fresh Lentinula edodes (LE) coated with polysaccharides (LEP) isolated from LE stalks and stored at 4 °C for 15 days. The results showed that compared to the chitosan-coated and uncoated LE, the LEP-treated mushrooms showed significant improvements in several qualities during storage, such as reduced weight loss, retention of hardness and springiness, improved soluble protein content, and reduced browning, malondialdehyde content, and electrolyte leakage rate. The best results were obtained with 1.5 % LEP. LEP improved the activities of peroxidase, catalase, superoxide dismutase, ascorbate peroxidase, and phenylalanine ammonialyase and significantly reduced the accumulation of hydrogen peroxide during storage compared to the control samples. In addition, the LEP treatment maintained the high antioxidant activity of LE during storage. Notably, LEP inhibited browning-related enzymes (polyphenol oxidase and tyrosinase) to reduce browning. It also maintained high levels of cellulase, chitinase, and β-1,3 glucanase to improve softening during storage. These findings suggest the potential of LEP to improve the post-harvest quality of mushrooms, allowing a storage period of up to 15 days (extending the shelf life by six days) and indirectly suggesting that the polysaccharide component of LEP can act as a self-defense additive to protect against spoilage during storage.

The metaxenia effects of different pollen grains on secondary metabolites enzymes and sugars of ‘Piarom’ date palm fruit

In this research, the characteristics of pollen were studied in eight pollinating cultivars of date palm, namely, ‘Shahani’, ‘Kabkab’, ‘Zahedi’, ‘Beraem’, ‘Faryab’, ‘Sheikhali’, ‘Fard’ and ‘Jarvis’. The characteristics were compared and metaxenia effects were evaluated on secondary metabolites, enzymes and other biochemical compounds of ‘Piarom’ date fruits. The evaluations were carried out during four stages of fruit growth and development. The pollen of these eight pollinating cultivars were compared in terms of carbohydrates, proteins, starch, total phenol, flavonoids, pectin methyl esterase, and amylase enzymes. According to the results, the pollen of ‘Sheikhali’, ‘Fard’, ‘Zahedi’ and ‘Shahani’ cultivars contained more of the above compounds, compared to the other cultivars. Regarding the effects of pollen on the composition of ‘Piarom’ date fruits, ‘Fard’ and ‘Sheikhali’ pollen produced the lowest amount of soluble tannin, which resulted in a better quality of ‘Piarom’ date fruits. Pollen was also obtained from ‘Sheikhali’ and ‘Fard’ cultivars for evaluations, showing that they led to the highest amounts of glucose and fructose in the fruits. Regarding the sucrose amount, ‘Jarvis’ and ‘Shikhali’ produced the best results. Pollen of ‘Sheikhali’ and ‘Fard’ cultivars caused the lowest amount of chlorophyll at the different stages of fruit growth, indicating a better decomposition of fruit chlorophyll and, as a result, better fruit quality. Pollen of ‘Sheikhali’ and ‘Fard’ cultivars produced the highest amounts of secondary metabolites such as total phenol, carotenoids and anthocyanin at the different stages of fruit development. The pollen of ‘Fard’ and ‘Sheikhali’ cultivars produced the highest levels of polygalacturonase, cellulase and invertase enzymes at different growth stages of the ‘Piarom’ date fruit. Regarding cellulase enzyme, fruits of the ‘Zahedi’ cultivar had more cellulase than the fruits of ‘Sheikhali’. In general, the pollen of ‘Fard’ and ‘Sheikhali, in comparison with other cultivars, improved the quantity and quality of ‘Piarom’ date fruits, due to their metaxenia properties.

Fusarium graminearum as a producer of xylanases with low cellulases when grown on wheat bran

The xylanolytic potential of endophytic fungi isolated from leaves of Theobroma cacao was explored for the first time. Four fungal strains showed significant amounts of xylanase activity and low cellulase levels when grown on wheat bran as the sole carbon source. Strain Ec220 of Fusarium graminearum had the highest xylanase production (1.79 U/ml), whereas its cellulase activity was minimal (0.24 U/ml). Optimal conditions for xylanase production were: 154 h of incubation time, pH 5.79 and 29.8 °C. Furthermore, two protein spots detected by two-dimensional gel electrophoresis showed molecular weights (26.05 and 27.70 kDa) and isoelectric points (6.18 and 9.20) corresponding to previously reported F. graminearum xylanases, Xyl A and Xyl B, respectively. Therefore, endophytic fungi of T. cacao can be an important source of xylanolytic activities when cultured on wheat bran, and xylanases with low cellulases found in strain Ec220 require further characterization as they show promise for possible industrial applications.

Ultrastructural Observations of Botrytis cinerea and Physical Changes in Resistant and Susceptible Grapevines.

The necrotrophic fungus Botrytis cinerea is a major threat to grapevine cultivation worldwide. Here, a highly resistant Chinese wild grapevine, Vitis amurensis 'Shuangyou' (SY), and the susceptible V. vinifera 'Red Globe' (RG) were selected for study, and their pathogenic infection and biochemical responses to B. cinerea were evaluated. The results revealed more trichomes on and a thicker cuticle for leaves of SY than RG under scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Both SEM and TEM also showed that conidial germination, appressorium formation, and hyphal development of B. cinerea were delayed on the leaves of resistant SY. Fewer infected hyphae were also observed in leaves of resistant SY when compared with susceptible RG. The infected leaves of resistant SY harbored higher levels of cellulase and pectinase activity during the early infection stages of B. cinerea at 4 h postinoculation (hpi), and higher glucanase and chitinase activity were maintained in the inoculated leaves of SY from 4 through 18 hpi. Lignin was deposited in the infected leaves of susceptible RG but not in resistant SY. Taken together, these results provide insights into the ultrastructural characterizations and physical changes in resistant and susceptible grapevines.

Effect of seed-borne fungi on seed health of native populations of Iranian cumin (Cuminum cyminum L.)

Seed borne diseases caused by fungal pathogens are one of the most important threats that reduce the quantity and quality of cumin production every year all over the world. Hence, the first step in this research was to identify the seed-borne fungi obtained from seeds of native populations of Iranian cumin (Cuminum cyminum L.), their effect on germination and vigor indices, the location of fungi in seeds and their transmission into seedlings. Based on morphological and molecular characters, the fungi were identified as Alternaria alternata, A. burnsii, Fusarium oxysporum and F. solani. Most of the identified isolates were present in the seed coat, while a few isolates of A. alternata was located in embryos. The results revealed that a negative correlation were found between incidences of naturally fungi infected seed, germination, and vigor indices. The pathogenicity test revealed that 20% of the isolates were non-pathogenic and other isolates were pathogenic or weakly pathogenic. In addition, the percentage seed transmission of fungal phytopathogens to cumin seedlings revealed that A. alternata was the more frequently isolated (67.27%) followed by F. oxysporum (52.80%), A. burnsii (32.74%) and F. solani (20.01%). Then, we investigated correlation among the activities of cell wall degrading enzymes (CWDEs) produced by fungal isolates and variation in pathogenicity and aggressiveness in vivo. Different levels of pathogenicity and aggressiveness were observed for various isolates of Fusarium and Alternaria species. Analysis of CWDEs activities, including cellulase, xylanase, pectinase and lipase showed that cellulase and lipase activities were more important than xylanase and pectinase activities for the pathogenicity of isolates. Therefore, these findings suggested that activity levels of cellulase and lipase are correlated with variation in pathogenicity and aggressiveness of seed-borne fungal isolates on seedling. This is the first report on isolation and identification of seed-borne fungi infection of cumin, together with investigating association between pathogenesis and CWDEs production by isolates isolated from seeds of Iranian cumin native populations. It is also the first time that the location of fungi in seeds and its seed-transmitted into cumin seedlings have been identified and evaluated, respectively.

Comparative evaluation of graded levels of untreated and cellulase and pectinase hydrolyzed corncob based diets on performance, carcass yield, haematological and biochemical parameters of broiler chickens

This study was conducted to compare the effects of feeding graded levels of untreated and cellulase and pectinase hydrolyzed corncob based diets on growth, nutrient digestibility, carcass yield, haematological and biochemical parameters of broiler chickens. Three hundred and fifteen day-old broiler chicks with an average initial body weight of 60g were used for the study. Seven dietary treatments were formulated such that Diet 1 was a control. Diets 2, 3 and 4 contained 5%, 10%, and 15% levels of untreated corncob, while diets 5, 6 and 7 contained cellulase and pectinase hydrolyzed corncob at 5%, 10% and 15% levels respectively. The birds were randomly distributed into seven treatment groups in three replicates and each replicate had fifteen birds. Crushed corncobs were pretreated with NaOH to remove the lignin contents and later subjected to locally produced cellulase and pectinase enzymes for the degradation of non-starch polysaccharides (NSPs) for five days. Proximate analyses of untreated corncobs, cellulase and pectinase hydrolyzed corncob, standard diet, diets containing untreated corncobs and diets containing cellulase and pectinase treated corncobs were carried out to determine the nutrient composition of the ingredients and feeds. Data obtained from Growth performance, nutrient digestibility, haematological and serum biochemical profile as well as carcass yield were determined in a 2x4 factorial arrangement. Feeding regime was twice daily for all the treatment groups while diets and water were supplied to birds ad libitum. The results showed that at the end of week eight when the experiment was terminated, the broilers placed on untreated corncob diets consumed higher quantities (2497.28 g) of feed than birds placed on enzyme treated diets (2142.94 g) and conversely recorded lower performance indices in all parameters measured. The result also showed that as the levels of untreated corncob diets increased, the final body weight of birds (1450.00 g) significantly (P&lt;0.05) reduced when compared to 2385.33g obtained for enzyme treated birds. The result equally showed that broilers fed hydrolyzed corncob diets had significantly higher (P&lt;0.05) and better performance in terms of feed conversion efficiency (0.67 as compared to 0.50) in the untreated birds, protein efficiency ratio (4.17 as compared to 1.69) in the untreated birds, feed conversion ratio (1.21 as compared to 2.70) in the untreated birds, nutrient digestibility, final body weight, dressed carcass percentage (80.22 as compared to 73.93%) in the untreated birds and carcass cuts when compared to broilers fed untreated corncob diets. Enzyme treated corncob diets significantly (P&lt; 0.05) lowered the blood cholesterol levels (1.85 g/dL as compared to 2.15 g/dL) in the untreated birds and also improved other haematological indices measured in the birds compared to the control broilers. Generally, broilers fed enzyme hydrolyzed corncob diets performed better with birds fed 15.00% enzyme treated corncob diets having the highest and the best performance record which could be attributed to better utilization of nutrients since the enzymes have hydrolyzed the nutrient releasing monomers and hence are better metabolized. It is concluded that feeding broilers with enzyme hydrolyzed corn cob at the levels 5 to15.0 % improved performance and carcass values. In addition, it elevated 99 albumin, globulin, total protein, Aspartate amino transaminase (AST) and Alanine amino transaminase (ALT) levels in the blood.

Production and characterization of cellulases derived from saprophytic fungi Penicillium bilaiae InaCC F16

Cellulases are the enzymes that hydrolyze β-1,4 glycosidic bonds in cellulose molecules into simpler molecules such as glucose. The cellulase complexes derived from microbes have been considered as a potential source for biochemical, biotechnology, and bioindustry. Thus, they are broadly used in industries for leather tanning, food, medicine, and detergent formulations. The important group of cellulase-producing fungi, such as Penicillium, is reportedly able to produce high levels of cellulase and hemicellulase complexes. This study aimed to determine the characterization and activity of cellulase enzymes produced by fungi Penicillium and analyze their cellulase activities, both qualitatively and quantitatively. The fungal strain used in this study was obtained from the Indonesian Culture Collection (InaCC) with the accession number of InaCC F15, InaCC F16, and InaCC F17. The parameters used for investigating the enzyme characteristic and activity were incubation time, index of substrate concentration on carboxymethyl cellulose (CMC), temperature, pH, and the effect of the addition of several metal ions as activators and/or inhibitors. The semi-qualitative analysis showed that the clear zone was formed in the surrounding colony of Penicillium InaCC F16 with a cellulolytic index was at 3.3. The optimum cellulase activity was obtained on two days incubation period, the temperature of 40°C, pH 5, and substrate concentration at 2%. The addition of metal ions at a concentration of 1mM showed that enzymes were activated by cations CoCl2, CuCl2, MgCl2, and inhibited by NaCl and ZnCl2.

Role of lignin and thermophilic lignocellulolytic bacteria in the evolution of humification indices and enzymatic activities during compost production

This study aimed to evaluate the effect of lignin content and thermophilic lignocellulolytic bacteria bioaugmentation on composting process. Treatments including bioaugmentation with thermophilic lignocellulolytic bacteria isolates such as Paenibacillus validus, Paenibacillus koreensis, Bacillus nealsonii, a mixture of the three mentioned bacterial isolates and control were compared at two level of organic media (high lignin content and low lignin content) in the form of nested factorial design. Several indices such as humification and enzymatic activities were monitored to evaluate the composting rate. The results revealed that high lignin treatments displayed higher ligninase, xylanase, protease and urease enzymatic activities compared to low lignin treatments. On the other hand, low lignin treatments showed higher level of humification indices, cellulase, beta-glucosidase and alkaline phosphomonoesterase enzymatic activities in comparison with high lignin treatments. Also, all measured enzymatic activities are at their highest between the second and the tenth weeks; however, this trend decreased to reach a steady point from the 18th weeks to the 24th weeks, but for urease enzymatic activity, a totally different trend in high and low lignin treatments was observed. Moreover, the highest humification indices as well as the cellulase and β-glucosidase enzymatic activities were associated to the Bacillus nealsonii isolate and the full consortium. They also displayed the highest ligninase, xylanase, protease, and urease and phosphatase activities. The efficient isolates shortened the time required for completing the composting process for about 2 to 4 weeks compared to the control treatments. For all measured indices, the control treatment had the lowest values.