Svetlana Zivanovic

Morphological and Surface Properties of Electrospun Chitosan Nanofibers

Nonwoven fiber mats of chitosan with potential applications in air and water filtration were successfully made by electrospinning of chitosan and poly(ethyleneoxide) (PEO) blend solutions. Electrospinning of pure chitosan was hindered by its limited solubility in aqueous acids and high degree of inter- and intrachain hydrogen bonding. Nanometer-sized fibers with fiber diameter as low as 80 +/- 35 nm without bead defects were made by electrospinning high molecular weight chitosan/PEO (95:5) blends. Fiber formation was characterized by fiber shape and size and was found to be strongly governed by the polymer molecular weight, blend ratios, polymer concentration, choice of solvent, and degree of deacetylation of chitosan. Weight fractions of polymers in the electrospun nonwoven fibers mats were determined by thermal gravimetric analysis and were similar to ratio of polymers in the blend solution. Surface properties of fiber mats were determined by measuring the binding efficiency of toxic heavy metal ions like chromium, and they were found to be related with fiber composition and structure.

Efficient Reduction of Chitosan Molecular Weight by High-Intensity Ultrasound : Underlying Mechanism and Effect of Process Parameters

The degradation of chitosan by high-intensity ultrasound (HIU) as affected by ultrasound parameters and solution properties was investigated by gel permeation chromatography coupled with static light scattering. The molecular weight, radius of gyration, and polydispersity of chitosan were reduced by ultrasound treatment, whereas chitosan remained in the same random coil conformation and the degree of acetylation did not change after sonication. The results demonstrate that (1) the degradation of chitosan by ultrasound is primarily driven by mechanical forces and the degradation mechanism can be described by a random scission model; (2) the degradation rate is proportional to M w (3); and (3) the degradation rate coefficient is affected by ultrasound intensity, solution temperature, polymer concentration, and ionic strength, whereas acid concentration has little effect. Additionally, the data indicate that the degradation rate coefficient is affected by the degree of acetylation of chitosan and independent of the initial molecular weight.

Molecular weight of chitosan influences antimicrobial activity in oil-in-water emulsions.

The objective of this study was to evaluate the antimicrobial efficiency of chitosans in oil-in-water emulsions. Model emulsions were prepared with 20% corn oil, 1.5% Tween 20, 1.5% Trypticase soy broth, 0.58% acetic acid, and chitosan polysaccharide or chitosan oligosaccharide in concentrations of 0, 0.1, 0.2, 0.5, and 0.7%. A control containing HCl was included to determine the role of acetic acid in the overall antibacterial activity. The pH of samples and controls was adjusted to 4.5. Emulsions were inoculated with Listeria monocytogenes (strains Scott A and 310) or Salmonella Typhimurium DT104 (strains 2486 and 2576) at a level of 10(7) CFU/ml. Inoculated emulsions were incubated at 10 and 25 degrees C for 4 days and analyzed for bacterial count every 24 h. Both tested Salmonella strains were more susceptible to acetic acid than Listeria. However, L. monocytogenes was more affected by chitosan than either Salmonella strain. During the storage at 25 degrees C, initial inoculum in the emulsions with 0.58% acetic acid and 0.1% chitosan polysaccharide was reduced to below the detection limits after 24, 48, 72, or 96 h for L. monocytogenes 310, Salmonella Typhimurium DT104 2576, Salmonella Typhimurium DT104 2486, or L. monocytogenes Scott A, respectively. Chitosan oligosaccharide was less effective against all tested bacteria and showed a concentration-dependent effect. The antimicrobial efficacy of chitosan was reduced at 10 degrees C, and reduction of microbial loads was delayed for approximately 24 h compared with 25 degrees C. Results suggest that addition of 0.1% chitosan polysaccharide would be sufficient to ensure the microbial safety of oil-in-water emulsions regardless of storage temperature.

Application of Chitosan Films Enriched with Oregano Essential Oil on Bologna – Active Compounds and Sensory Attributes:

Chitosan films prepared with oregano essential oil were applied on bologna slices. Release of the essential oil compounds during film preparation and application on the meat product and consumer acceptability of bologna enriched with oregano essential oil were tested. Oregano essential oil compounds were quantified by gas chromatography mass spectroscopy (GCMS) after extraction from the filmforming solution, films before and after application on bologna and from bologna slices before and after application of the films. The results indicated that the concentration of components of the essential oil sharply decreased during film preparation, e.g. from 757.7 ppm carvacrol in film-forming solution to 2.1 ppm in dried films. No carvacrol was detected in the films after application on bologna for 5 days at 4°C, mainly due to its diffusion into bologna. It seemed that the moisture and high lipid content of bologna helped the diffusion of the oregano essential oil from the chitosan film matrix into the product. Sensory evaluation suggested that addition of 45 ppm or less of oregano oil to bologna would be acceptable to consumers. Results support the potential use of chitosan–oregano essential oil films as an antimicrobial packaging material for processed meat.

Inactivation of Escherichia coli K-12 in Apple Juice Using Combination of High-Pressure Homogenization and Chitosan

Apple juice and apple cider were inoculated with Escherichia coli K-12 and processed using a high-pressure homogenizer to study bacterial inactivation. Seven levels of pressure ranging from 50 to 350 MPa were used in the high-pressure homogenizer. Two types of chitosan (regular and water soluble) with 2 levels of concentration 0.01% and 0.1% were investigated for synergistic effect with high-pressure homogenization for the bacterial inactivation. E. coli K-12 inactivation was evaluated as a function of homogenizing pressure at different concentration of 2 types of chitosan in apple juice and cider. High-pressure homogenization (HPH) induced significant inactivation in the range of 100 to 200 MPa, while thermal inactivation was the primary factor for the bacterial inactivation above 250 MPa. Significant (P < 0.05) 2-way interactions involving pressure and type of substrate or pressure and chitosan concentration were observed during the study. The homogenization pressure and the incremental quantity of chitosan (both types) acted synergistically with the pressure to give higher inactivation. Significantly (P < 0.05) higher inactivation was observed in apple juice than apple cider at same homogenizing pressure. No effect of type of chitosan was observed on the bacterial inactivation.

Effect of chitosan on the infectivity of murine norovirus, feline calicivirus, and bacteriophage MS2.

Chitosan is known to inhibit microorganisms of concern to plants, animals, and humans. However, the effect of chitosan on human enteric viruses of public health concern has not been extensively investigated. The purpose of this study was to determine the effect of chitosan on three human enteric viral surrogates: murine norovirus 1 (MNV-1), feline calicivirus F-9 (FCV-F9), and (ssRNA) bacteriophage MS2 (MS2). Chitosan oligosaccharide lactate (molecular weight of 5,000) and water-soluble chitosan (molecular weight of 53,000) at concentrations of 1.4, 0.7, and 0.35% were incubated at 37 degrees C for 3 h with equal volumes of each virus at high (approximately 7 log PFU/ml) and low (approximately 5 log PFU/ml) titers. Chitosan effects on each treated virus were evaluated with standardized plaque assays in comparison to untreated virus controls. The water-soluble chitosan at 0.7% decreased the FCV-F9 titer by approximately 2.83 log PFU/ml, with decreasing effects at lower concentrations, and also decreased MS2 at high titers by approximately 1.18 to 1.41 log PFU/ml, regardless of the concentration used. Chitosan treatments at the concentrations studied had no effect on MNV-1 at high titers. Chitosan oligosaccharide showed similar trends against the viruses, but to a lesser extent compared with that of water-soluble chitosan. When lower virus titers (approximately 5 log PFU/ml) were used, plaque reduction was observed for FCV-F9 and MS2, but not MNV-1. The use of higher-molecular-weight chitosan and at higher concentrations with longer incubation may be necessary to inactivate MNV-1. These results in the plaque reduction of human enteric virus surrogates by chitosan treatment show promise for its potential application in the food environment.

Inactivation of human enteric virus surrogates by high-intensity ultrasound.

Foodborne viruses, especially human noroviruses, are recognized as leading causes of nonbacterial gastroenteritis worldwide. Development of effective inactivation methods is of great importance to control their spread. In this study, the effect of high-intensity ultrasound (HIUS) on the infectivity of three foodborne virus surrogates was investigated. The three surrogates, murine norovirus (MNV-1), feline calicivirus (FCV-F9), and MS2 bacteriophage, were diluted in phosphate-buffered saline (PBS) or orange juice to a titer of approximately 6 log(10) PFU/mL or approximately 4 log(10) PFU/mL. The ultrasound treatment was performed in duplicate by immersing the HIUS probe in virus-containing solution that was cooled in ice-water and sonicated at 20 kHz for 2, 5, 10, 15, 20, and 30 min with 30 sec on and 30 sec off. The infectivity of the recovered viruses after each ultrasound treatment was evaluated in duplicate using standardized plaque assays and compared to untreated controls. The results show that HIUS effectiveness depended on the virus type, the initial titer of the viruses, and the virus suspension solution. At titers of approximately 4 log(10) PFU/mL in PBS, feline calicivirus (FCV)-F9, MS2, and murine norovirus (MNV)-1 required 5-, 10-, and 30-min treatment, respectively, for complete inactivation. At initial titers of approximately 4 log(10) PFU/mL in orange juice, FCV-F9 required a 15-min treatment for complete inactivation and only a 1.55 log(10) PFU/mL reduction was achieved for MNV-1 in orange juice after 30-min treatment. Thus, inactivation by HIUS in orange juice was much lower than in PBS. Experiments using titers of approximately 6 log(10) PFU/mL showed decreased effects compared to those using titers of approximately 4 log(10) PFU/mL. These results indicate that HIUS alone is not sufficient to inactivate virus in food. Hurdle technologies that combine HIUS with antimicrobials, heat, or pressure should be explored for viral inactivation.

Effectiveness of chitosan on the inactivation of enteric viral surrogates

Chitosan is known to have bactericidal and antifungal activity. Although human noroviruses are the leading cause of non-bacterial gastroenteritis, information on the efficacy of chitosan against foodborne viruses is very limited. The objective of this work was to determine the effectiveness of different molecular weight chitosans against the cultivable human norovirus and enteric virus surrogates, feline calicivirus, FCV-F9, murine norovirus, MNV-1, and bacteriophages, MS2 and phiX174. Five purified chitosans (53, 222, 307, 421, ~1150 kDa) were dissolved in water, 1% acetic acid, or aqueous HCl pH = 4.3, sterilized by membrane filtration, and mixed with equal volume of virus to obtain a final concentration of 0.7% chitosan and 5 log(10) PFU/ml virus. Virus-chitosan suspensions were incubated for 3 h at 37 °C. Untreated viruses in PBS, in PBS with acetic acid, and in PBS with HCl were tested as controls. Each experiment was run in duplicate and replicated at least twice. Water-soluble chitosan (53 kDa) reduced phiX174, MS2, FCV-F9 and MNV-1 titers by 0.59, 2.44, 3.36, and 0.34 log(10) PFU/ml respectively. Chitosans in acetic acid decreased phiX174 by 1.19-1.29, MS2 by 1.88-5.37, FCV-F9 by 2.27-2.94, and MNV-1 by 0.09-0.28 log(10) PFU/ml, respectively. Increasing the MW of chitosan corresponded with an increasing antiviral effect on MS2, but did not appear to play a role for the other three tested viral surrogates. Overall, chitosan treatments showed the greatest reduction for FCV-F9, and MS2 followed by phiX174, and with no significant effect on MNV-1.

Composition and Bioactive Properties of Yerba Mate (llex paraguariensis A. St.-Hil.): A Review

Yerba mate es una infusion popular producida y consumida en Argentina, Brasil, Chile, Paraguay y Uruguay. Se procesa a partir de hojas y tallos de Ilex paraguariensis A. St.-Hil., un arbusto perenne de la familia Aquifoliaceae. El procesamiento ocurre en seis etapas: recoleccion de hojas maduras y tallos pequenos, tostado por fuego directo, secado por aire caliente, molienda, envejecimiento (dependiendo de los atributos sensoriales requeridos), y embalaje final. Si bien la yerba mate se ha cultivado y consumido por siglos en America del Sur, su popularidad en los Estados Unidos ha aumentando debido a la demanda por bebidas saludables y alimentos mas naturales y por los potenciales beneficios para la salud de la yerba mate (antioxidante, antimicrobiano, accion contra la obesidad y diabetes, digestivo, estimulante). La yerba mate tambien se ha investigado como agente de prevencion y causa de algunos tipos de cancer, causando controversia entre investigadores. Investigaciones recientes han ampliado el espectro de uso de la yerba mate como agente antimicrobiano, proteccion de cultivos y accion contra patogenos transmitidos por alimentos. Resultados prometedores para el uso de esta planta en la salud humana y animal han llevado a esta revision. Esta revision se centro en la composicion de la yerba mate, y el efecto que el cultivo y el procesamiento puede tener sobre sus propiedades.

The effect of solvent composition on grafting gallic acid onto chitosan via carbodiimide

The primary antioxidant (AOX) activity of chitosan can be introduced by grafting of phenolic compound - gallic acid (GA) to its amino and/or hydroxyl groups. The objective of this study was to investigate the effect of ethanol (EtOH) concentration (0%, 25%, 50%, and 75% in water) on efficiency of grafting GA onto chitosan in the presence of 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC)/N-hydroxysuccinimide (NHS). The grafting was confirmed by FTIR and the efficiency was quantified as Folins total phenolics. When pure deionized water was used as a sole solvent (0% EtOH), GA was grafted to chitosan at the largest extent (285.9mg GA/g chitosan). As the concentration of EtOH increased, the grafting efficiency proportionally decreased. NMR studies showed that EtOH inhibited grafting of GA by prohibiting the production of the intermediate - NHS ester. The results confirm that the concentration of EtOH in grafting solution significantly affects grafting efficiency of GA on chitosan.