Monitoring monkeypox virus in saliva and air samples in Spain: a cross-sectional study.

BACKGROUND: The transmission of monkeypox virus occurs through direct contact, but transmission through saliva or exhaled droplets and aerosols has not yet been investigated. We aimed to assess the presence of monkeypox virus DNA and infectious virus in saliva samples and droplets and aerosols exhaled from patients infected with monkeypox virus. METHODS: We did a cross-sectional study in patients with monkeypox confirmed by PCR who attended two health centres in Madrid, Spain. For each patient, we collected samples of saliva, exhaled droplets within a mask, and aerosols captured by air filtration through newly developed nanofiber filters. We evaluated the presence of monkeypox virus in the samples by viral DNA detection by quantitative PCR (qPCR) and isolation of infectious viruses in cell cultures. FINDINGS: Between May 18 and July 15, 2022, 44 patients with symptomatic monkeypox attended two health centres in Madrid and were included in the study. All were cisgender men, with a median age of 35·0 years (IQR 11·3). We identified high loads of monkeypox virus DNA by qPCR in 35 (85%) of 41 saliva samples. Infectious monkeypox virus was recovered from 22 (67%) of 33 saliva samples positive for monkeypox virus DNA. We also found a significant association between the number of affected cutaneous areas or general symptoms and the viral load present in saliva samples. Droplets exhaled from patients with monkeypox, detected inside a mask, contained monkeypox virus DNA in 32 (71%) of 45 samples, with two of the 32 positive samples showing the presence of the infectious virus. Monkeypox virus DNA in aerosols, collected from the medical consultation room, were detected in 27 (64%) of 42 samples, despite patients wearing an FFP2 mask during the visit. Infectious virus was not recovered from aerosol samples. High levels of monkeypox virus DNA were identified in aerosols collected from a hospital isolation room housing a patient with monkeypox. INTERPRETATION: The identification of high viable monkeypox virus loads in saliva in most patients with monkeypox and the finding of monkeypox virus DNA in droplets and aerosols warrants further epidemiological studies to evaluate the potential relevance of the respiratory route of infection in the 2022 monkeypox virus outbreak. FUNDING: EU, Consejo Superior de Investigaciones Científicas, and Ciberinfec.

Development of an Electrospun Patch Platform Technology for the Delivery of Carvedilol in the Oral Mucosa.

The work herein presented aims to develop and characterize carvedilol (CVD) releasable non-water-soluble monolayers and a multilayer patch made of ultrathin micron and submicron fibers for drug delivery into the sublingual mucosa. Firstly, the developed formulations containing CVD within different biopolymers (PDLA, PCL, and PHB) were characterized by scanning electron microscopy (SEM), attenuated total reflectance Fourier transformed infrared spectroscopy (ATR-FTIR), differential scanning calorimetry (DSC), wide-angle X-ray scattering (WAXS), and for their in vitro drug release. SEM micrographs assessed the fiber morphology attained by adding carvedilol. ATR-FTIR spectra revealed good chemical compatibility between CVD and the tested biopolymers, whereas DSC and WAXS confirmed that CVD was in an amorphous state within the biopolymeric fibers. In vitro release studies showed enhanced CVD release kinetics from the electrospun biopolymer monolayers compared to the dissolution rate of the commercial form of the pure drug, except for the slow-releasing PDLA fibers. Finally, the selected CVD-loaded layer, i.e., electrospun PHB, was built into a three-layer patch to tackle mucosa adhesion and unidirectional release, while retaining the enhanced release kinetics. The patch design proposed here further demonstrates the potential of the electro-hydrodynamic processing technology to render unique mucoadhesive controlled delivery platforms for poorly water-soluble drugs.

Photothermal Activatable Mucoadhesive Fiber Mats for On-Demand Delivery of Insulin via Buccal and Corneal Mucosa.

Electrospun fiber mats loaded with therapeutics have gained considerable attention as a versatile tool in the biomedical field. While these bandages are largely based on fast-dissolving polymers to release the cargo, stimuli-responsive fiber mats have the advantages of providing a timely and spatially controlled drug delivery platform, which can be refilled and reused several times. These benefits make electrospun fiber patches original platforms for painless and convenient on-demand hormone release. Because of the high need of more convenient and non-invasive methods for delivering insulin, a hormone that is currently used to treat hundred million people with diabetes worldwide, we have investigated the tremendous potential of reduced graphene oxide modified poly(acrylic acid) based fiber mats as an original platform for buccal and corneal insulin delivery on-demand. The PAA@rGO hydrogel-like fibers rendered water-insoluble by incorporating ß-cyclodextrin, followed by thermal cross-linking, which showed adequate tensile strength along with high adsorption capacity of insulin at pH 7 and good recyclability. The fiber mats maintained good fibrous morphology and high loading efficiency even after five loading-release cycles. The mucoadhesive nature of the fibers allowed their application for insulin delivery via the eye cornea and the buccal mouth lining, as evidenced in ex vivo studies. Insulin loaded PAA@rGO hydrogel-like fibers showed an insulin flux via buccal lining of pigs of 16.6 ± 2.9 µg cm-2 h-1 and 24.3 ± 3.1 µg cm-2 h-1 for porcine cornea. Testing on healthy adult volunteers confirmed the excellent, mucoadhesive nature of the bandage, with three out of six volunteers feeling completely comfortable (note 8.3) while wearing the patches in the buccal cavity.

Formulation of the biological control yeast Meyerozyma caribbica by electrospraying process: effect on postharvest control of anthracnose in mango (Mangifera indica L.) and papaya (Carica papaya L.).

BACKGROUND: Microorganism for biological control of fruit diseases is an eco-friendly alternative to the use of chemical fungicides. RESULTS: This is the first study evaluating the electrospraying process to encapsulate the biocontrol yeast Meyerozyma caribbica. The effect of encapsulating material [Wey protein concentrate (WPC), Fibersol® and Trehalose], its concentration and storage temperature on the cell viability of M. caribbica, and in vitro and in vivo control of Colletotrichum gloeosporioides was evaluated. The processing with commercial resistant maltodextrin (Fibersol®) 30% (w/v) as encapsulating material showed the highest initial cell viability (95.97 ± 1.01%). The storage at 4 ± 1 °C showed lower losses of viability compared to 25 ± 1 °C. Finally, the encapsulated yeast with Fibersol 30% w/v showed inhibitory activity against anthracnose in the in vitro and in vivo tests, similar to yeast fresh cells. CONCLUSION: Electrospraying was a highly efficient process due to the high cell viability, and consequently, a low quantity of capsules is required for the postharvest treatment of fruits. Additionally, the yeast retained its antagonistic power during storage. © 2021 Society of Chemical Industry.

Development and Characterization of Electrospun Biopapers of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Derived from Cheese Whey with Varying 3-Hydroxyvalerate Contents.

In the present study, three different newly developed copolymers of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) with 20, 40, and 60 mol % contents in 3-hydroxyvalerate (3HV) were produced by the biotechnological process of mixed microbial cultures (MMCs) using cheese whey (CW), a by-product from the dairy industry, as feedstock. The CW-derived PHBV copolyesters were first purified and then processed by solution electrospinning, yielding fibers of approximately 2 µm in cross-section in all cases. The resultant electrospun PHBV mats were, thereafter, post-processed by annealing at different temperatures, below their maximum of melting, selected according to their 3HV content in order to obtain continuous films based on coalesced fibers, so-called biopapers. The resultant PHBV films were characterized in terms of their morphology, crystallinity, and mechanical and barrier properties to assess their potential application in food packaging. The CW-derived PHBV biopapers showed high contact transparency but a slightly yellow color. The fibers of the 20 mol % 3HV copolymer were seen to contain mostly poly(3-hydroxybutyrate) (PHB) crystals, the fibers of the 40 mol % 3HV copolymer a mixture of PHB and poly(3-hydroxyvalerate) (PHV) crystals and lowest crystallinity, and the fibers of the 60 mol % 3HV sample were mostly made of PHV crystals. To understand the interfiber coalesce process undergone by the materials during annealing, the crystalline morphology was also assessed by variable-temperature both combined small-angle and wide-angle X-ray scattering synchrotron and Fourier transform infrared experiments. From these experiments and, different from previously reported biopapers with lower 3HV contents, all samples were inferred to have a surface energy reduction mechanism for interfiber coalescence during annealing, which is thought to be activated by a temperature-induced decrease in molecular order. Due to their reduced crystallinity and molecular order, the CW-derived PHBV biopapers, especially the 40 mol % 3HV sample, were found to be more ductile and tougher. In terms of barrier properties, the three copolymers performed similarly to water and limonene, but to oxygen, the 40 mol % sample showed the highest relative permeability. Overall, the materials developed, which are compatible with the Circular Bioeconomy organic recycling strategy, can have an excellent potential as barrier interlayers or coatings of application interest in food packaging.

High-Oxygen-Barrier Multilayer Films Based on Polyhydroxyalkanoates and Cellulose Nanocrystals.

This study reports on the development and characterization of organic recyclable high-oxygen-barrier multilayer films based on different commercial polyhydroxyalkanoate (PHA) materials, including a blend with commercial poly(butylene adipate-co-terephthalate) (PBAT), which contained an inner layer of cellulose nanocrystals (CNCs) and an electrospun hot-tack adhesive layer of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) derived from cheese whey (CW). As a result, the full multilayer structures were made from bio-based and/or compostable materials. A characterization of the produced films was carried out in terms of morphological, optical, mechanical, and barrier properties with respect to water vapor, limonene, and oxygen. Results indicate that the multilayer films exhibited a good interlayer adhesion and contact transparency. The stiffness of the multilayers was generally improved upon incorporation of the CNC interlayer, whereas the enhanced elasticity of the blend was reduced to some extent in the multilayer with CNCs, but this was still much higher than for the neat PHAs. In terms of barrier properties, it was found that 1 µm of the CNC interlayer was able to reduce the oxygen permeance between 71% and 86%, while retaining the moisture and aroma barrier of the control materials.

Blends of Poly(3-Hydroxybutyrate-co-3-Hydroxyvalerate) with Fruit Pulp Biowaste Derived Poly(3-Hydroxybutyrate-co-3-Hydroxyvalerate-co-3-Hydroxyhexanoate) for Organic Recycling Food Packaging.

In the present study, a new poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate) [P(3HB-co-3HV-co-3HHx)] terpolyester with approximately 68 mol% of 3-hydroxybutyrate (3HB), 17 mol% of 3-hydroxyvalerate (3HV), and 15 mol% of 3-hydroxyhexanoate (3HHx) was obtained via the mixed microbial culture (MMC) technology using fruit pulps as feedstock, a processing by-product of the juice industry. After extraction and purification performed in a single step, the P(3HB-co-3HV-co-3HHx) powder was melt-mixed, for the first time, in contents of 10, 25, and 50 wt% with commercial poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV). Thereafter, the resultant doughs were thermo-compressed to obtain highly miscible films with good optical properties, which can be of interest in rigid and semirigid organic recyclable food packaging applications. The results showed that the developed blends exhibited a progressively lower melting enthalpy with increasing the incorporation of P(3HB-co-3HV-co-3HHx), but retained the PHB crystalline morphology, albeit with an inferred lower crystalline density. Moreover, all the melt-mixed blends were thermally stable up to nearly 240 °C. As the content of terpolymer increased in the blends, the mechanical response of their films showed a brittle-to-ductile transition. On the other hand, the permeabilities to water vapor, oxygen, and, more notably, limonene were seen to increase. On the overall, this study demonstrates the value of using industrial biowaste derived P(3HB-co-3HV-co-3HHx) terpolyesters as potentially cost-effective and sustainable plasticizing additives to balance the physical properties of organic recyclable polyhydroxyalkanoate (PHA)-based food packaging materials.

Study of the Compatibilization Effect of Different Reactive Agents in PHB/Natural Fiber-Based Composites.

Fiber-matrix interfacial adhesion is one of the key factors governing the final properties of natural fiber-based polymer composites. In this work, four extrusion reactive agents were tested as potential compatibilizers in polyhydroxylbutyrate (PHB)/cellulose composites: dicumyl peroxide (DCP), hexamethylene diisocyanate (HMDI), resorcinol diglycidyl ether (RDGE), and triglycidyl isocyanurate (TGIC). The influence of the fibers and the different reactive agents on the mechanical properties, physical aging, and crystallization behavior were assessed. To evaluate the compatibilization effectiveness of each reactive agent, highly purified commercial cellulose fibers (TC90) were used as reference filler. Then, the influence of fiber purity on the compatibilization effect of the reactive agent HMDI was evaluated using untreated (U_RH) and chemically purified (T_RH) rice husk fibers, comparing the results with the ones using TC90 fibers. The results show that reactive agents interact with the polymer matrix at different levels, but all compositions showed a drastic embrittlement due to the aging of PHB. No clear compatibilization effect was found using DCP, RDGE, or TGIC reactive agents. On the other hand, the fiber-polymer interfacial adhesion was enhanced with HMDI. The purity of the fiber played an important role in the effectiveness of HMDI as a compatibilizer, since composites with highly purified fibers showed the greatest improvements in tensile strength and the most favorable morphology. None of the reactive agents negatively affected the compostability of PHB. Finally, thermoformed trays with good mold reproducibility were successfully obtained for PHB/T_RH/HMDI composition.

Improved Process to Obtain Nanofibrillated Cellulose (CNF) Reinforced Starch Films with Upgraded Mechanical Properties and Barrier Character.

Nowadays, the interest on nanofibrillated cellulose (CNF) has increased owing to its sustainability and its capacity to improve mechanical and barrier properties of polymeric films. Moreover, this filler shows some drawbacks related with its high capacity to form aggregates, hindering its dispersion in the matrix. In this work, an improved procedure to optimize the dispersability of CNF in a thermoplastic starch was put forward. On the one hand, CNF needs a hydrophilic dispersant to be included in the matrix, and on the other, starch needs a hydrophilic plasticizer to obtain a thermoformable material. Glycerol was used to fulfil both targets at once. CNF was predispersed in the plasticizer before nanofibrillation and later on was included into starch, obtaining thin films. The tensile strength of these CNF-starch composite films was 60% higher than the plain thermoplastic starch at a very low 0.36% w/w percentage of CNF. The films showed a noticeable correlation between water uptake, and temperature and humidity. Regarding permeability, a ca. 55% oxygen and water vapor permeability drop was found by nanofiller loading. The hydrolytic susceptibility of the composite was confirmed, being similar to that of the thermoplastic starch.

Valorization of Municipal Biowaste into Electrospun Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Biopapers for Food Packaging Applications.

The present study reports on the production and characterization of a new biopackaging material made of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) derived from municipal biowaste (MBW) and produced by the mixed bacterial culture technology. After purification and extraction, the MBW-derived PHBV was processed by electrospinning to yield defect-free ultrathin fibers, which were thermally post-treated. Annealing at 130 °C, well below the biopolymer's melting temperature (Tm), successfully yielded a continuous film resulting from coalescence of the electrospun fibrillar morphology, the so-called biopaper, exhibiting enhanced optical and color properties compared to traditional melt compounding routes. The crystallinity and crystalline morphology were comprehensively studied as a function of temperature by attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy and combined time-resolved synchrotron small- and wide-angle X-ray scattering (SAXS and WAXS) experiments, which clearly indicated that the molecular order within the copolyester was improved up to a maximum at 130 °C, and then it decreased at the biopolymer's Tm. It was hypothesized that by annealing at the temperature at which the thermally induced molecular order is maximized, the fibers generated sufficient mobility to align alongside, hence reducing surface energy and porosity. The data suggest that this material shows a good balance between enhanced mechanical and improved barrier properties to vapors and gases in comparison to traditional paper and other currently used petroleum-derived polymers, thus presenting significant potential to be part of innovative food biopackaging designs for the protection and preservation of foods in a circular bioeconomy scenario.

Physicochemical, Antioxidant and Antimicrobial Properties of Electrospun Poly(ε-caprolactone) Films Containing a Solid Dispersion of Sage (Salvia officinalis L.) Extract.

In this study, novel active films made of poly(ε-caprolactone) (PCL) containing a solid dispersion of sage extract (SE) were developed by means of the electrospinning technique and subsequent annealing treatment. Initially, the antioxidant and antimicrobial potential of SE was confirmed. Thereafter, the effect of SE incorporation at different loading contents (5%, 10%, and 20%) on the physicochemical and functional properties of the films was evaluated. The films were characterized in terms of morphology, transparency, water contact angle, thermal stability, tensile properties, water vapor, and aroma barrier performances, as well as antioxidant and antimicrobial activities. Thin, hydrophobic films with good contact transparency were produced by annealing of the ultrathin electrospun fibers. Interestingly, the effect of SE addition on tensile properties and thermal stability of the films was negligible. In general, the water vapor and aroma permeability of the PCL-based films increased by adding SE to the polymer. Nevertheless, a strong 2,2-diphenyl-1-picrylhydrazyl (DPPH·) free radical scavenging ability, and a strong activity against foodborne pathogens Staphylococcus aureus and Escherichia coli were achieved by SE incorporation into PCL matrix. Overall, the obtained results suggest great potential of the here-developed PCL-based films containing SE in active food packaging applications with the role of preventing oxidation processes and microbial growth.

Antibacterial and Barrier Properties of Gelatin Coated by Electrospun Polycaprolactone Ultrathin Fibers Containing Black Pepper Oleoresin of Interest in Active Food Biopackaging Applications.

The present study evaluated the effect of using electrospun polycaprolactone (PCL) as a barrier coating and black pepper oleoresin (OR) as a natural extract on the morphology, thermal, mechanical, antimicrobial, oxygen, and water vapor barrier properties of solvent cast gelatin (GEL). The antimicrobial activity of the developed multilayer system obtained by the so-called electrospinning coating technique was also evaluated against Staphylococcus aureus strains for 10 days. The results showed that the multilayer system containing PCL and OR increased the thermal resistance, elongated the GEL film, and significantly diminished its permeance to water vapor. Active multilayer systems stored in hermetically closed bottles increased their antimicrobial activity after 10 days by inhibiting the growth of Staphylococcus aureus. This study demonstrates that addition of electrospun PCL ultrathin fibers and OR improved the properties of GEL films, which promoted its potential use in active food packaging applications.

On the Use of the Electrospinning Coating Technique to Produce Antimicrobial Polyhydroxyalkanoate Materials Containing In Situ-Stabilized Silver Nanoparticles.

Electro-hydrodynamic processing, comprising electrospraying and electrospinning techniques, has emerged as a versatile technology to produce nanostructured fiber-based and particle-based materials. In this work, an antimicrobial active multilayer system comprising a commercial polyhydroxyalkanoate substrate (PHA) and an electrospun PHA coating containing in situ-stabilized silver nanoparticles (AgNPs) was successfully developed and characterized in terms of morphology, thermal, mechanical, and barrier properties. The obtained materials reduced the bacterial population of Salmonella enterica below the detection limits at very low silver loading of 0.002 ± 0.0005 wt %. As a result, this study provides an innovative route to generate fully renewable and biodegradable materials that could prevent microbial outbreaks in food packages and food contact surfaces.

Hybrid encapsulation structures based on ß-carotene-loaded nanoliposomes within electrospun fibers.

Hybrid encapsulation structures based on ß-carotene-loaded nanoliposomes incorporated within the polymeric ultrathin fibers produced through electrospinning were developed to improve the photostability of the antioxidant. These novel materials were intended to incorporate ß-carotene into water-based food formulations, overcoming the existing limitations associated with its hydrophobic character. Initially, both empty and antioxidant-loaded nanoliposomes were developed and incorporated into polyvinyl alcohol (PVOH) and polyethylene oxide (PEO) solutions. The changes in the solution properties were evaluated to determine their effects on the electrospinning processing. The mixed polymer solutions were subsequently electrospun to produce hybrid nanoliposome-loaded ultrathin fibers. FTIR analysis confirmed the presence of phospholipid molecules inside the electrospun fibers. These ultrathin fibers were evaluated regarding their morphology, diameter, internal ß-carotene distribution and stability against UV irradiation. Liposomal release studies from the electrospun fibers were also undertaken, confirming the presence of the liposomal structures after dissolving the electrospun fibers in water.

Antimicrobial beeswax coated polylactide films with silver control release capacity.

Although the application of silver based antimicrobial systems is a widespread technology, its implementation in areas such as food packaging is still challenging. The present paper describes the fabrication of poly(lactic acid) (PLA) coated with beeswax with controlled release properties for sustained antimicrobial performance. Release of silver ions from the polymers was monitored voltammetrically under various conditions (surface contact, immersion in various liquid media and at different pH values) throughout at least 7days. A higher release was noted with decreasing pH while surface release was much slower than the release when immersed in liquid medium. While uncoated films demonstrated a high burst release which in some instances implied surpassing some current migration restrictions (<0.05mg/kg food), the addition of a beeswax layer allowed a sustained release of the antimicrobial compound. Increasing the thickness of the beeswax layer resulted in an increase in the water barrier properties of the films while reducing the relatively constant values of sustained release. Antimicrobial performance was correlated with the release of silver ions, indicating threshold concentrations for biocide action of <6µg/L and 9-14µg/L for surface contact and in liquid media, respectively. Either by surface contact or by immersion in growth medium or vegetable soup, the coated films displayed a strong bactericidal effect against Salmonella enterica. The application of this functional barrier thus offers the possibility of tuning the release profiles of the films to suit a specific application and puts forth the possible suitability of these materials for food packaging or other migration sensitive applications.

Morphology and water barrier properties of nanobiocomposites of κ/ι-hybrid carrageenan and cellulose nanowhiskers.

The current study presents the development and characterization of novel carrageenan nanobiocomposites showing enhanced water barrier due to incorporation of cellulose nanowhiskers (CNW). CNW, prepared by acid hydrolysis of highly purified α cellulose microfibers, were seen to have a length of around 25-50 nm and a cross section of ca. 5 nm when dispersed in the matrix. The nanobiocomposites were prepared by incorporating 1, 3, and 5 wt % of the CNW into a carrageenan matrix using a solution casting method. Morphological data (TEM and optical microscopy) of the nanocomposites containing CNW were compared with the morphology of the corresponding biocomposites containing the original cellulose microfibers and the differences discussed. Thermal stability by TGA, water vapor permeability, and percent water uptake were also determined. The main conclusion arising from the analysis of the results is that the nanobiocomposites containing 3 wt % of CNW exhibited the lowest reduction in water vapor permeability, that is, ca. 71%, and that this reduction was largely attributed to a filler-induced water solubility reduction. This fully biobased nanoreinforced carrageenan can open new opportunities for the application of this biopolymer in food-packaging and -coating applications.