Medium Molecular Weight Chitosan Research Articles

Pectin-chitosan hydrogels with modified properties for the encapsulation of strawberry phenolic compounds

Pectin-chitosan hydrogels with blends of low (50–190 kDa) and medium (310–395 KDa) molecular weight (MW) chitosan (LC and MC, respectively) were developed, and their characteristics were investigated before and after the encapsulation of an aqueous strawberry extract. The pectin to total chitosan mass ratio, the composition of the strawberry extract and the MW of chitosan greatly affected the interactions between pectin and chitosan at different pH values. More specifically, blends of low and medium MW chitosan improved the stability of the strawberry-gels in acidic conditions compared to their corresponding MC-gels, showed better flow and texture profiles, as well as slower release of phenolic compounds during in vitro digestion compared to the only stable LC-gel. Therefore, by manipulating the length range of chitosan chains would allow the formation of pectin-chitosan hydrogels with improved properties for the development of functional food products.

Effect of medium molecular weight chitosan coating on quality of houndfish (Tylosurus crocodilus) during ice storage

Effect of medium molecular weight chitosan coating on quality of houndfish (Tylosurus crocodilus) during ice storage

Chitosan-based mucoadhesive films loaded with curcumin for topical treatment of oral cancer

This study aimed to develop mucoadhesive chitosan-based films capable of enhancing the curcumin penetration into the oral mucosa to treat oral cancers. We developed three films containing medium molecular weight chitosan (190–310 KDa) and other excipients (polyvinyl alcohol, Poloxamer®407, and propylene glycol) that have proven to be compatible with each other and with curcumin in thermal analyses. The films were smooth, flexible, and precipitates free, with uniform weight and thickness, pH compatible with the oral mucosa, resistance to traction, and entrapped curcumin in a high proportion. They also exhibited necessary swelling and mucoadhesion for tissue adherence. Ex vivo penetration studies proved that the films significantly increased the penetration of curcumin into the oral mucosa compared to control, even when the mucosa was subjected to a condition of simulated salivation. Curcumin exhibited cytotoxic activity in vitro in the two head and neck cancer cell lines (FaDu, SCC-9) at doses close to those found in penetration studies with the films. When combined with radiotherapy, curcumin demonstrated superiority over single doses of radiotherapy at 4, 8, and 12 Gy. Therefore, the developed films may represent a promising alternative for the topical treatment of oral tumors.

Development of a chitosan/propolis-based polymeric system: Characterization, biocompatibility, and modulation of transcription factor expression

Chitosan (CHT) and propolis (P) are natural materials with antiseptic, anti-inflammatory, antimicrobial, and mucoadhesive properties with many applications in orthopaedia dentistry. The objective of this study was to develop a low molecular weight (LMW) and medium molecular weight (MMW) chitosan polymeric system with propolis at different concentrations (0.31%, 0.62%, 1.25%, 2.5%, and 5%) and to evaluate the effect in terms of their cell interaction with dental pulp fibroblasts. FTIR spectra of both molecular weights CHT’s films with propolis show increased transmittance bands at 1730–1720 cm−1 and 1460 cm−1, indicating the presence of propolis ethanolic extract in the organic matrix. Contact angles test to determine hydrophobicity CHT films functionalized with propolis exhibit a more hydrophobic surface as propolis concentration increases. Cytotoxicity showed synergy between chitosan and propolis; the highest proliferation was observed in LMW CHT with 0.31% of propolis. Membranes made with CHT of both molecular weights plus propolis in an inflammatory stage (cells previously treated with LPS) can inhibit the expression of NF-k-B, MAPK, and VEGF. A polymeric system based on chitosan/ethanolic extract propolis could be a future alternative treatment for oral ulcers.

An Investigation into the Effects of Processing Factors on the Properties and Scaling-Up Potential of Propranolol-Loaded Chitosan Nanogels.

Chitosan-triphosphate (TPP) nanogels are widely studied drug delivery carrier systems, typically prepared via a simple mixing process. However, the effects of the processing factors on nanogel production have not been extensively explored, despite the importance of understanding and standardising such factors to allow upscaling and commercial usage. This study aims to systematically evaluate the effects of various fabrication and processing factors on the properties of nanogels using a Design of Experiment approach. Hydrodynamic size, polydispersity index (PDI), zeta potential, and encapsulation efficiency were determined as the dependent factors. The temperature, stirring rate, chitosan grade, crosslinker choice, and the interaction term between temperature and chitosan grade were found to have a significant effect on the particle size, whereas the effect of temperature and the addition rate of crosslinker on the PDI was also noteworthy. Moreover, the addition rate of the crosslinker and the volume of the reaction vessel were found to impact the encapsulation efficiency. The zeta potential of the nanogels was found to be governed by the chitosan grade. The optimal fabrication conditions for the development of medium molecular weight chitosan and TPP nanogels included the following: the addition rate for TPP solution was set at 2 mL/min, while the solution was then stirred at a temperature of 50 °C and a stirring speed of 600 rpm. The volume of the glass vial used was 28 mL, while the stirrer size was 20 mm. The second aim of the study was to evaluate the potential for scaling up the nanogels. Size and PDI were found to increase from 128 nm to 151 nm and from 0.232 to 0.267, respectively, when the volume of the reaction mixture was increased from 4 to 20 mL and other processing factors were kept unchanged. These results indicate that caution is required when scaling up as the nanogel properties may be significantly altered with an increasing production scale.

Antibacterial wound coating based on chitosan and povidone, obtained by 3D printing

The objective of this study was to develop a method for forming an antimicrobial wound coating based on chitosan and polyvinylpyrrolidone using 3D printing technology.
 The properties of the coating were then studied in vitro and in vivo to improve the treatment outcomes of deep burns. The resulting coating was a 4% hydrogel of medium molecular weight chitosan with the addition of 1% povidone iodine and dermal fibroblasts. After transplantation, the coating was covered with “Foliderm” film. The coating was formed using an extrusion 3D bioprinter, with printing parameters determined experimentally. The samples were first studied in vitro. Scanning electron microscopy was used to evaluate the coating’s microarchitecture and its interaction with dermal fibroblasts. A colorimetric test was conducted to assess cell metabolic activity and cytotoxicity, and antimicrobial activity against reference strains of Staphylococcus aureus was analyzed. An experiment was conducted to evaluate the in vivo properties of the coating. Nineteen male Wistar rats were used in the study. An injury was inflicted that resulted in a deep thermal contact burn, affecting all layers of skin and subcutaneous fatty tissue, with an area of approximately 20 cm2. The animals were divided into three groups: experimental (with the application of the developed coating), comparative (using the traditional and widespread method of treatment with Levomekol ointment) and control (without treatment).
 The study lasted for 38 days and found that the developed coating is highly biocompatible, atraumatic, elastic, and adheres well to wounds. Chitosan was used to create a porous structure with channels running parallel to each other. The coating cells are evenly distributed on the surface of the matrix, specifically on the walls of the pores. The inclusion of 1% povidone iodine in the polymer resulted in high antimicrobial activity without significantly affecting the activity of the cells in the composition. The experiment on applying a coating for treating deep thermal burns demonstrated that the developed coating had a positive effect on the wound healing process. This effect was characterized by a higher rate of epithelization and a significantly lower incidence of infectious complications compared to other experimental groups. In the histological study, the experimental group outperformed the control and comparison groups in the quality of the formed granulation tissue, the number of newly formed capillaries, and the severity of the local inflammatory process.

The Potential of Chitosan-Based Composites for Adsorption of Diarrheic Shellfish Toxins.

Okadaic acid (OA) is one of the most potent marine biotoxins, causing diarrheal shellfish poisoning (DSP). The proliferation of microalgae that produce OA and its analogues is frequent, threatening human health and socioeconomic development. Several methods have been tested to remove this biotoxin from aquatic systems, yet none has proven enough efficacy to solve the problem. In this work, we synthesized and characterized low-cost composites and tested their efficacy for OA adsorption in saltwater. For the synthesis of the composites, the following starting materials were considered: chitosan of low and medium molecular weight (CH-LW and CH-MW, respectively), activated carbon (AC), and montmorillonite (MMT). Characterization by vibrational spectroscopy (FTIR), X-ray diffraction (XRD), and microscopy revealed differences in the mode of interaction of CH-LW and CH-MW with AC and MMT, suggesting that the interaction of CH-MW with MMT has mainly occurred on the surface of the clay particles and no sufficient intercalation of CH-MW into the MMT interlayers took place. Among the composites tested (CH-LW/AC, CH-MW/AC, CH-MW/AC/MMT, and CH-MW/MMT), CH-MW/MMT was the one that revealed lower OA adsorption efficiency, given the findings evidenced by the structural characterization. On the contrary, the CH-MW/AC composite revealed the highest average percentage of OA adsorption (53 ± 11%). Although preliminary, the results obtained in this work open up good perspectives for the use of this type of composite material as an adsorbent in the removal of OA from marine environments.

Physicochemical and kinetic properties of collagenase, native and adsorbed on chitosan

One of the modern research areas in biotechnology and medicine is the development of highly stable biopreparations based on immobilized enzymes. Using the method of sorption immobilization, heterogeneous biocatalysts were obtained based on collagenase (EC 3.4.24.3) from Clostridium histolyticum and from medium molecular weight (200 kDa) and high molecular weight (350 kDa) chitosans. The goal of the work was to study the physicochemical and kinetic properties of free and immobilized collagenase from Clostridium histolyticum on a chitosan matrix. The physicochemical and kinetic properties of the enzyme were studied in soluble and immobilized states using classical biophysical and biochemical methods. It was shown that the maximum activity of native collagenase was observed at 37ºС, while in case of the enzyme immobilized on medium and high molecular weight chitosan it was observed at 40ºС. Sorption immobilization of the biocatalyst on a chitosan matrix allowed increasing its stability during incubation within the temperature range of 40-70°C. After incubation at 80 and 90°C, no significant differences in the activity of soluble and immobilized collagenase were found. When studying the dependence of the protease activity of the enzyme on the medium acidity in the pH range from 5.0 to 9.0, it was established that the maximum catalytic ability of soluble collagenase was observed at a pH value of 7.5. The biocatalyst immobilized on medium and high molecular weight chitosans was highly active at pH 6.0-7.5. The catalytic ability of the enzyme in free and in adsorbed states noticeably decreased at pH 8.0, while at pH 9.0 soluble collagenase immobilized on medium and high molecular weight chitosans was completely inactive. The maximum rate of the enzymatic reaction of native and chitosan-immobilized biocatalysts was observed at a substrate concentration of 0.04 mM. Apparent values of V´max и K´m were calculated. The immobilization of collagenase on medium and high molecular weight chitosans did not result in a decrease of the maximum reaction rate or a change in the degree of affinity of the enzyme to the substrate. Using dynamic light scattering, we confirmed that the structure of collagenase in solution was most stable at pH 7.0-8.0, as exactly in this range of [H+] values we discovered the highest content of particles corresponding to the size of native collagenase and the lowest content of its aggregates. The optimal temperature for the best stability value of collagenase (no autolysis processes and minimal particle aggregation) can be considered as being 46°C.

The Effect of Chitosan Incorporation on Physico-Mechanical and Biological Characteristics of a Calcium Silicate Filling Material.

A tricalcium silicate-based cement, Biodentine™, has displayed antibiofilm activity when mixed with chitosan powder. This study aimed to assess the effect of chitosan incorporation on the physico-mechanical and biological properties of Biodentine™. In this study, medium molecular weight chitosan powder was incorporated into Biodentine™ in varying proportions (2.5 wt%, 5 wt%, 10 wt%, and 20 wt%). The setting time was determined using a Vicat apparatus, solubility was assessed by calculating weight variation after water immersion, radiopacity was evaluated and expressed in millimeters of aluminum, the compressive strength was evaluated using an Instron testing machine, and the microhardness was measured with a Vickers microhardness tester. In addition, surface topography of specimens was analyzed using scanning electron microscopy, and the effect of chitosan on the viability of human embryonic kidney (HEK 293) cells was measured by a colorimetric MTT assay. Incorporation of 2.5 wt% and 5 wt% chitosan powder delivered an advantage by speeding up the setting time of Biodentine material. However, the incorporation of chitosan compromised all other material properties and the crystalline structure in a dose-dependent manner. The chitosan-modified material also showed significant decreases in the proliferation of the HEK 293 cells, signifying decreased biocompatibility. Chitosan incorporation into calcium silicate materials adversely affects the physical and biological properties of the material. Despite the increased antimicrobial activity of the modified material, the diminution in these properties is likely to reduce its clinical value.

Thermosensitive composite based on agarose and chitosan saturated with carbon dioxide. Preliminary study of requirements for production of new CSAG bioink

This study introduces a method for producing printable, thermosensitive bioink formulated from agarose (AG) and carbon dioxide-saturated chitosan (CS) hydrogels. The research identified medium molecular weight chitosan as optimal for bioink production, with a preferred chitosan hydrogel content of 40–60 %. Rheological analysis reveals the bioink's pseudoplastic behavior and a sol-gel phase transition between 27.0 and 31.5 °C. The MMW chitosan-based bioink showed also the most stable extrusion characteristic. The choice of chitosan for the production of bioink was also based on the assessment of the antimicrobial activity of the polymer as a function of its molecular weight and the degree of deacetylation, noting significant cell reduction rates for E. coli and S. aureus of 1.72 and 0.54 for optimal bioink composition, respectively. Cytotoxicity assessments via MTT and LDH tests confirm the bioink's safety for L929, HaCaT, and 46BR.1 N cell lines. Additionally, XTT proliferation assay proved the stimulating effect of the bioink on the proliferation of 46BR.1 N fibroblasts, comparable to that observed with Fetal Bovine Serum (FBS). FTIR spectroscopy confirms the bioink as a physical polymer blend. In conclusion, the CS/AG bioink demonstrates the promising potential for advanced spatial cell cultures in tissue engineering applications including skin regeneration.

Comparative study of the effect of cross‐linking degree on chitosan hydrogels synthesized with low and medium molecular weight chitosan

AbstractChitosan (CS) is often used in hydrogels due to its natural origin, sustainability, non‐toxicity, and biodegradability. However, the physicochemical properties of CS hydrogels generated via physical cross‐linking are relatively poor. CS hydrogels were prepared by cross‐linking with glutaraldehyde (GA) at different ratios ranging from 1% to 10% (w/w), using two different molecular weight CS. The swelling degree of hydrogels was investigated at pH 2.0, 5.6, and 7.4. The structure, morphology, and thermal characteristics of hydrogels were examined using Fourier transform infrared, scanning electron microscopy, differential scanning calorimetry, and thermogravimetric analysis, respectively. Compression tests were conducted using Zwick/Roell universal testing equipment following ASTM D695. In vitro degradation studies were performed in enzyme and enzyme‐free mediums at pH 7.4. The study demonstrated that low molecular weight CS (L‐CS) hydrogels have a lower sol content and higher swelling values than medium molecular weight CS (M‐CS) hydrogels. Both L‐CS and M‐CS hydrogels cross‐linked with 1% GA showed maximum swelling degrees at pH 2.0. Thermal analysis concluded that M‐CS hydrogels have higher thermal stability than L‐CS hydrogels. It was also found that the M‐CS hydrogels have higher compressive moduli. The degradation experiments in both enzyme and enzyme‐free media showed that L‐CS hydrogels have higher degradation rates.Highlights Chemically cross‐linked hydrogels synthesized using low and medium molecular weight chitosan. The mechanical and physical properties of medium and low molecular weight chitosan hydrogels were compared. Degradation experiments were performed in both enzyme and enzyme‐free media. Swelling values decreased noticeably with the increasing molecular weight. Low molecular weight chitosan improved the biodegradability of hydrogels.

Effects of chitosan with different molecular weights on storage quality and fungi inhibition of mini-cucumber

The effect of low (LWC, 30 kD), medium (MWC, 150 kD) and high molecular weight chitosan (HWC, 300 kD) coatings on storage quality and fungi inhibition of mini-cucumber was evaluated. Chitosan coatings significantly reduced the weight loss, and delayed the color change and sensory score of mini-cucumber when compared with the control. Chitosan coatings significantly retarded the decrease of DPPH free radical scavenging rate and total phenol content, and inhibited the total number of bacterial colonies and the viable numbers of yeast and molds in mini-cucumber. Chitosan coatings also significantly reduced the disease incidence and disease severity of mini-cucumber artificial inoculated with Botrytis cinerea. LWC coating showed superior preservation effectiveness and antifungal activity to MWC and HWC coating. Compared with the control, LWC coating prolonged the shelf life by 6 days which was evaluated by sensory score, and reduced the total number of bacterial colonies and viable numbers of yeast and molds in mini-cucumber by 4.20 log10 CFU/g and 2.72 log10 CFU/g, respectively. These results suggested that chitosan molecular weights affected preservation effectiveness and antifungal activity against B. cinerea and LWC could be used to prolong the shelf life of mini-cucumber and maintain its quality after harvest.

Characterization of alginate from Antarctic Himantothallus grandifolius (Phaeophyceae) and preparation of polyelectrolyte complexes with chitosan

AbstractHydrogels are three‐dimensional polymeric materials with applications in various areas such as biomedicine, tissue engineering, pollutant capture systems, and so forth. However, its application is limited due to the reproducibility of its properties. Factors such as the composition of the constituents, the type of union between the polymer chains, and the crosslinking time play a crucial role in ensuring its reproducibility. In this study, the objective is to prepare a matrix with well‐defined properties that ensure controlled release processes. Treatment of Himantothallus grandifolius with Na2CO3 aqueous solution afforded in 18.9% yield alkaline extract. Hydrogels were prepared by ionic complexation of alginate from H. grandifolius, and polyguluronate enriched alginate from Desmarestia ligulata with chitosan stabilized by immersion in 0.1 M CaCl2 solution; better swelling ratio in water was found for hydrogels with low molecular weight chitosan (MW 125,000 g/mol) than those with medium molecular weight chitosan (MW450,000–500,000 g/mol). Liberation of the model drug metformin hydrochloride from alginates: chitosan complexes (3:1) at pH 1.5 was especially low for hydrogel from H. grandifolius during the first 2 h; 70% of metformin was released at pH 6.5 after 24 h. This polyelectrolyte complex constitutes a good material for potential application in biomedicine considering its cationic and anionic characteristics.

Application of low-, and medium-molecular weight chitosan for preparation of spray-dried microparticles loaded with Ferulago angulata essential oil: Physicochemical, antioxidant, antibacterial and in-vitro release properties

The present work aimed at spray-drying encapsulation of Chavir (Ferulago angulata) essential oil (EO) using low-, and medium-molecular weight chitosan. The obtained EO was observed to be mainly composed of β-ocimene, α-pinene, and bornyl acetate with antioxidant, and antimicrobial activity. The results indicated that stable emulsions with uniform particle size distribution and encapsulation efficiencies higher than 93 % could be prepared using chitosan as feed for spray-drying. In addition, spray-drying resulted in fabricating stable microspheres with yields higher than 50 %, uniform particle size, and encapsulation efficiency exceeding 70 %. The microspheres were fairly soluble and hygroscopic, and exhibited antioxidant and bacteriostatic activities with a biphasic release pattern. FTIR characterisation confirmed successful encapsulation of EO and thermal properties of microspheres indicated enhanced stability of EO after microencapsulation. Overall, it was revealed that molecular weight of chitosan and EO:chitosan ratio affects some physicochemical properties of obtained chitosan microspheres.

Synthesized carboxymethyl-chitosan variant composites for cyclic adsorption-desorption based removal of Fe, Pb, and Cu

The global issue of environmental contamination from industrial wastewater comprising Cu, Fe and Pb demands effective treatment strategies. In this article, a functional composite sorbent was devised to selectively remove copper, iron, and lead from a real-world mimicking wastewater system. For the purpose, high, medium, and low molecular weight chitosan with amine and hydroxyl functional groups were used as a substrate, and glutaraldehyde was used to anchor the organic compound with carboxymethyl groups. Characterization with X-ray diffraction, Fourier transform infrared spectroscopy, thermogravimetric analysis, X-ray analyzer accompanied by field emission scanning electron microscope, and Brunauer-Emmett-Teller were conducted for the synthesized adsorbent. Accordingly, the properties of the adsorbent were evaluated to infer that the synthesis assured a purified and functionalized system. The surface area of the medium carboxymethyl chitosan derivative was analyzed as 31.43 m2 g−1. Various adsorption parameters were examined methodically to assess upon optimal removal requirements. The effect of adsorbent dosage, contact time and concentration on the adsorption of the studied metal ions were conducted and the optimum values were achieved at pH 3.82, 540 min contact duration and 1.2 g L−1 sorbent dose. Maximum adsorbent capacities of 344.83 mg g−1, 9.59 mg g−1, and 90.09 mg g−1 were realized for Cu, Pb, and Fe, respectively. The experimental measurements of the studied heavy metal ions inferred the best fitness of Langmuir isotherm equilibrium and pseudo second order kinetic models. Further, elution studies with easy-to-deploy low-cost acidic and basic eluents (HCl, HNO3, H2SO4, KOH, and NaOH) were conducted with cyclic adsorption-desorption strategies. These investigations confirmed the adsorbent's good reusability up to 3 cycles of adsorption and its proximity to serve as a potential material for multi-heavy metal ions elimination from complex adsorbate systems.

Chitosan: Properties and Its Application in Agriculture in Context of Molecular Weight.

Chitosan is a naturally occurring compound that can be obtained from deacetylated chitin, which is obtained from various sources such as fungi, crustaceans, and insects. Commercially, chitosan is produced from crustaceans. Based on the range of its molecular weight, chitosan can be classified into three different types, namely, high molecular weight chitosan (HMWC, >700 kDa), medium molecular weight chitosan (MMWC, 150-700 kDa), and low molecular weight chitosan (LMWC, less than 150 kDa). Chitosan shows several properties that can be applied in horticultural crops, such as plant root growth enhancer, antimicrobial, antifungal, and antiviral activities. Nevertheless, these properties depend on its molecular weight (MW) and acetylation degree (DD). Therefore, this article seeks to extensively review the properties of chitosan applied in the agricultural sector, classifying them in relation to chitosan's MW, and its use as a material for sustainable agriculture.

Comparison of shrimp waste-derived chitosan produced through conventional and microwave-assisted extraction processes: Physicochemical properties and antibacterial activity assessment

Depending on its physicochemical properties and antibacterial activities, chitosan can have a wide range of applications in food, pharmaceutical, medicine, cosmetics, agriculture, and aquaculture. In this experimental study, chitosan was extracted from shrimp waste through conventional extraction, microwave-assisted extraction, and conventional extraction under microwave process conditions. The effects of the heating source on the physicochemical properties and antibacterial activity were investigated. The results showed that the heating process parameters affected the physicochemical properties considerably. The conventional procedure yielded high molecular weight chitosan with a 12.7 % yield, while the microwave extraction procedure yielded a porous medium molecular weight chitosan at 11.8 %. The conventional extraction under microwave process conditions led to medium molecular weight chitosan with the lowest yield (10.8 %) and crystallinity index (79 %). Antibacterial assessment findings revealed that the chitosan extracted using the conventional method had the best antibacterial activity in the agar disk diffusion assay against Listeria monocytogenes (9.48 mm), Escherichia coli. (8.79 mm), and Salmonella Typhimurium (8.57 mm). While the chitosan obtained by microwave-assisted extraction possessed the highest activity against E. coli. (8.37 mm), and Staphylococcus aureus (8.05 mm), with comparable antibacterial activity against S. Typhimurium (7.34 mm) and L. monocytogenes (6.52 mm). Moreover, the minimal inhibitory concentration and minimal bactericidal concentration assays demonstrated that among the chitosan samples investigated, the conventionally-extracted chitosan, followed by the chitosan extracted by microwave, had the best antibacterial activity against the target bacteria.

Biocatalysts based on papain associates with chitosan nanoparticles

The research purpose was to develop and study biocatalysts based on papain associates with chitosan nanoparticles. We obtained medium and high molecular weight chitosan nanoparticles, both with and without ascorbic acid . When the papainna-noparticles complexes with ascorbic acid were formed, the catalytic activity of the enzyme increased by 3 % for medium molecular weight chitosan and by 16 % for high molecular weight chitosan. After 168 hours of incubation in 0.05 M of Tris-HCl buffer (pH 7.5) at 37 °C, the free enzyme retained 15 % of its catalytic activity, whereas its associates with chitosan nanoparticles exhibited ~ 30 %. The papain complex with chitosan nanoparticles and ascorbic acid exhibited 40 % of the enzyme catalytic activity. We simulated the bonds and interactions within the chitosan-ascorbic acid-papain complex. The proposed biocatalysts have high prospects for effective use in cosmetology, biomedicine, and pharmacy.

Evaluation of antifungal efficacy of chitosan against Aspergillus fumigatus of stored rice (Oryza sativa)

Contamination of stored rice with fungal pathogens results in rice of poor quality and economic value, and may also have harmful effects on human and animal health. This study aimed at evaluating the antifungal efficacy of chitosan against Aspergillus fumigatus of stored rice (Oryza sativa). Four forms of chitosan were used in this study viz; Purchased Low, Medium and high molecular weight chitosan and chitosan synthesized from Crab Shell using deproteinization, demineralization, decolouration and deacetylation. Degree of deacetylation of the synthesized chitosan was determined using Fourier transform infrared (FTIR) analysis. The in vitro antifungal activity of all forms of chitosan were determined using food poisoning method against Aspergillus fumigatus isolated from stored rice seeds using agar plate methods. The percentage inhibition of mycelial radial growth Aspergillus fumigatus by the different forms of chitosan was determined. Fourier Transfrom InfraRed Spectra of synthesized chitosan showed major absorption bands from 3444.72, 2966.17, 2512.60, 2144.84, 1429.74, 1258.12, 1160.05, 1025.2, 869.92, 710.50, 608.40 to 559.36. Absorption peak at 559 was assigned to glucopyranose ring in the chitosan matrix. Degree of deacetylation of synthesized chitosan was at 98.6% degree. Highest mycelia radial growth of 8.00 cm was recorded in control, while 2.00, 0.00, 5.23, and 1.33cm were recorded in 2.0% concentration of High Molecular Weight Chitosan (HMWC), Medium Molecular Weight Chitosan (MMWC), Low Molecular Weight Chitosan (LMWC) and Chitosan synthesized from Crab shell (CSCS) treatment respectively. The result of the percentage inhibition reveals that A. fumigatus had 100% inhibition in 1.5% HMWC and 2.0% MMWC while 85% inhibition in 2.0% CSCS which were significantly different from 2.0% LMWC with 36.5% inhibitions. Results obtained from this study has shown the effectiveness of chitosan as an effective antifungal agent which can improve and maintain the quality of stored rice and ensure a safer environment due to its nontoxic and environmental friendly properties.

Cultivar-Specific Effect of Chitosan on Chitinase and Glucanase Activity in the Roots of Garlic Allium sativum L.

Chitosan is a natural polysaccharide and, when applied externally, is able to stimulate both growth and defense of the plant, enhancing its resistance to abiotic stresses and suppressing the development of many phytopathogens. Immune response includes the activation of defense proteins, carbohydrases such as chitinases and glucanases, which are also known to participate in the regulation of morphogenesis. In this study, for the first time, the effect of treatment with unfractionated (hydrolysate) chitosan of low (CH1) and medium (CH2) molecular weight on chitinase and glucanase activities, as well as on the expression of chitinase and -1,3-glucanase genes in the roots of two cultivars of garlic Allium sativum L. differing by resistance to Fusarium rot was examined. It was shown that the effect of chitosans on the enzymatic activity and expression of the genes encoding -1,3-glucanases (AsPR2a, AsPR2b, and AsPR2c) and chitinases (AsCHI1, AsCHI3, AsCHI7, AsCHI17, and AsCHI23) is cultivar-specific, which may be due to different susceptibility of the cultivars to Fusarium. The expression pattern of chitinase genes AsCHI10, AsCHI27, and AsCHI34, similar between varieties, suggested their involvement in root tissue morphogenesis. The results indicated a greater stimulatory effect of CH2 in comparison with CH1 on chitinase and glucanase activity. The stronger inhibitory influence of CH2 (as compared with CH1) on the expression of chitinase and -1,3-glucanase genes correlated with the lower fungicidal effect of CH2 on Fusarium proliferatum. The findings may be used in breeding biotechnology to increase the resistance of garlic to Fusarium.