Chitosan/starch beads as bioinoculants carrier: long-term survival of bacteria and plant growth promotion.

Immobilization of microorganisms in biodegradable polymeric matrices constitutes a promising technology for plant growth promoting to overcome the challenging conditions of the rhizosphere. Previously, we demonstrated that beads prepared from blends of chitosan/starch of analytical grades ionically cross-linked are useful carriers for Azospirillum brasilense and Pseudomonas fluorescens. The aims of this work were to study A. brasilense Az39 and P. fluorescens ZME4 immobilization in industrial quality beads produced with a blend of chitosan/starch, to assess bacterial survival during long-term storage and biofilm distribution in the beads. We also proposed to analyze the consortia root colonization and its performance as plant growth-promoting bioinoculants compared to liquid counterpart. Our results revealed that A. brasilense Az39 and P. fluorescens ZME4 can coexist in industrial grade chitosan/starch beads, and this mixed immobilization benefits the survival rates of both species, even for more than a year under shelf storage. Confocal laser scanning microscopy with fluorescent dyed strains showed that both species remain mainly in different locations inside and over the beads. Additionally, maize seed treatment with beads-loaded bacteria resulted in growth promotion of roots in a similar manner than traditional liquid-based inoculation. The evidence collected here demonstrate that low-cost chitosan/starch beads are a suitable carrier for bacteria consortia and could be a reliable alternative to liquid inoculation in agronomic practices with additional benefits for industrial management. KEY POINTS: • Mixed immobilization increases bacterial survival in chitosan/starch industrial beads • Beads increase competence of bacteria in rhizosphere of maize • Inoculation mediated by beads promotes plant growth of maize.

Chitosan-starch beads prepared by ionotropic gelation as potential matrices for controlled release of fertilizers.

The present study examines the agrochemical application of macrospheres prepared with chitosan and chitosan-starch blends by an easy dripping technique, using a sodium tripolyphosphate aqueous solution as the crosslinking agent. These biopolymers form hydrogels that could be a viable alternative method to obtain controlled-release fertilizers (CRFs). Three different concentrations (ranging from 20 to 100wt/wt% of chitosan) and two crosslinking times (2 or 4h) were used. The resulting polymeric matrices were examined by scanning electron microscopy coupled with energy dispersive X-ray, X-ray diffraction, Fourier transform infrared spectroscopy, solid-state nuclear magnetic resonance, thermogravimetric analysis and differential scanning calorimetry. Ionotropic gelation and neutralization induced the formation of the macrospheres. The crosslinking time and the composition of the polymeric hydrogel controlled the crosslinking degree, the swelling behavior and the fertilizer loading capability. Potassium nitrate-loaded beads were shown to be useful as a controlled-release fertilizer. After 14days of continuous release into distilled water, the cumulative concentration in the release medium reached between 70 and 93% of the initially loaded salt, depending on the matrix used. The prepared beads showed properties that make them suitable for use in the agrochemical industry as CRFs.

Sclerotium rolfsii scleroglucan: the promising behavior of a natural polysaccharide as a drug delivery vehicle, suspension stabilizer and emulsifier.

Gel matrices of scleroglucans from Sclerotium rolfsii ATCC 201126 (EPS I and EPS II, from 48-h and 72-h fermentations, respectively) were evaluated on their release kinetics of theophylline (Th). Equivalent polymer (2%, w/w) and Th (0.2%, w/w) concentrations showed almost coincident drug release patterns, independently of polymer molecular weight or the microstructural properties of gel matrices. Dynamic rheological studies of scleroglucan hydrogel structures (storage, G', and loss, G'', moduli) indicated a solid-like behavior. Differences on pore size dimensions (EPS I=20 microm and EPS II=7 microm) were in accordance to the differences in G' (EPS I=113 Pa and EPS II=161 Pa), a fact likely related to variations in the cross-linking density of polymer networks. Compared to already known biopolymers, EPS I and EPS II at 0.5 g/L showed a good dispersing ability against particulate suspensions of activated charcoal, bentonite, CaCO(3), celite and quartz powder. Emulsifying ability of both EPSs at 2g/L was high (E=56-60%) when tested with kerosene, moderate ( approximately 30%) with hexadecane, and negligible in the presence of olive oil-in-water emulsions.

Dynamic rheological measurements and drug release kinetics in swollen scleroglucan matrices.

The structure of scleroglucan gel matrices was characterized by dynamic rheological studies. The results were compared with the release kinetics of theophylline in analogous samples using a Franz diffusion cell, fitting the drug release data with a semi-empirical power law. Dynamic rheology gave information about the viscous and elastic components (loss and storage moduli, respectively) of the gel which could influence the drug-release profiles. Scleroglucan gels showed two structural transitions within the gel regime that coincided with changes in the release pattern. It was found that the introduction of 0.4% (w/w) of theophylline decreased the loss and storage moduli in the 2% (w/w) scleroglucan gels by 50%. The influence of the same wt.% theophylline in other gels was strongly dependent on the gel concentration. These results demonstrated the value of rheological studies to detect matrix structural changes produced by the inclusion of drugs which may modify the drug-release profile.

Correlation between gel structural properties and drug release pattern in scleroglucan matrices.

The drug delivery behavior of scleroglucan hydrogels was correlated with the structural features found by means of steady-state fluorescence studies. Theophylline at 0.4% in weight was used as the model drug and its release was measured using a Franz cell device. The results were fitted with a semi-empirical power law equation, finding significant differences between the 0.5% and higher than 0.8% scleroglucan concentrations (w/w). Four molecular fluorescent probes were used to test microenvironmental properties of the gel cavities, which will play a key role on the release of the pharmaceutical drugs. The results found by the fluorescence analysis are in good agreement with the ones obtained by release kinetics.