3D Bioprinting of Food Grade Hydrogel Infused with Living Pleurotus ostreatus Mycelium in Non-sterile Conditions.

Mycelium is the root-like network of fungi. Mycelium biocomposites prepared by template replication (molding) can function as environmentally friendly alternatives to conventional polystyrene foams, which are energy- and carbon-intensive to manufacture. Recently, several studies have shown that 3D bioprinting technologies can be used to produce high value functional mycelium products with intricate geometries that are otherwise difficult or impossible to achieve via template replication. A diverse range of nutrients, thickeners, and gelling agents can be combined to produce hydrogels suitable for 3D bioprinting. 3D bioprinting with hydrogel formulations infused with living fungi produces engineered living materials that continue to grow after bioprinting is complete. However, a hydrogel formulation optimized for intricate 3D bioprinting of Pleurotus ostreatus mycelium, which is among the strains most commonly used in mycelium biocomposite fabrication, has yet to be described. Here, we design and evaluate a versatile hydrogel formulation consisting of malt extract (nutrient), carboxymethylcellulose and cornstarch (thickeners), and agar (gelling agent), all of which are easily sourced food grade reagents. We also outline a reproducible workflow to infuse this hydrogel with P. ostreatus liquid culture for 3D bioprinting of intricate structures comprised of living P. ostreatus mycelium and characterize the changes in height and mass as well as hardness of the prints during mycelium growth. Finally, we demonstrate that the workflow does not require a sterile bioprinting environment to achieve successful prints and that the same mycelium-infused hydrogel can be supplemented with additives such as sawdust to produce mycelium biocomposite objects. These findings demonstrate that 3D bioprinting using mycelium-based feedstocks could be a promising biofabrication technique to produce engineered living materials for applications such as mushroom cultivation, food preparation, or construction of the built environment.

Asymmetrical Competition between Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae) Coexisting in Breeding Sites.

Aedes aegypti and Culex quinquefasciatus are mosquito vectors for several tropical diseases that represent a current public health problem. The ecological requirements for each species are different, however, both species show high biological adaptability, which promotes their coexistence in the same breeding sites. The purpose of this study was to assess the effect of larval association between Ae. aegypti and Cx. quinquefasciatus under different laboratory conditions of food supply and temperature, and under field simulated conditions like peridomestic containers. Our findings showed that under field simulated conditions there was no asymmetrical competition in mixed cultures with the different Cx. quinquefasciatus/Ae. aegypti ratios tested. However, under laboratory conditions in which different doses of food supply were evaluated, it was observed that competition between the two species takes place. Larval coexistence under food scarcity conditions (0.95 mg/larva) showed that Ae. aegypti had a greater adult emergence than Cx. quinquefasciatus and was capable of depriving Cx. quinquefasciatus of the food needed to complete metamorphosis. In an intermediate dose of food (1.9 mg/larva), the dry weight of Cx. quinquefasciatus adults decreased, and their larval development time increased when Cx. quinquefasciatus/Ae. aegypti ratio was low. Also, a temperature effect was assessed demonstrating that Cx. quinquefasciatus was more vulnerable to changes in temperature. We suggest that Ae. aegypti is more successful in exploiting microhabitats when food is scarce, due to its scrape active feeding habitats and fast larval development times. Therefore, in conditions of food paucity both species will compete, and Ae. aegypti larvae will prevail.