Mushrooms are added to traditional dishes including banosh and kulesha. The main components of banosh and kulesha are corn flour and polonynska bryndza . Both traditional dishes serve as a base to add either berries or mushrooms, depending on the holiday. Forest mushroom soup is also a very common first course and has long been a part of the Hutsul, traditional diet. During specific Christian holidays, fasting is a practice and “it is important for people to stock with dried mushrooms.” [Katya .] Mushroom hunting is embedded in Ukrainian culture overall but even more so in the Carpathian forests, where these mushrooms grow. The role of polonynas in Hutsul landscape is intertwined with traditional foods, specifically in the making of sheep’s cheese . Polonynska bryndza is made during the summer months and obtained from milk of local Carpathian sheep or cows. The process of making bryndza is at least a 600-year-old tradition, and is deeply intertwined with traditional food and landscape, specifically high meadows, called polonynas . This tradition, passed down from generation to generation, preserves ecocultural memories tied to plant and lichen species found in polonynas as well the process of making polonynska bryndza. As noted in the introduction, the decline of polonynas is linked to cattle population decline after the collapse of the Soviet Union, when keeping cattle became economically difficult and expensive. Due to this decline, it is synergistically changing the landscape and its floral diversity, leading to overgrowth. Without grazers and active tending of the land, this biocultural reservoir faces loss. The decline of livestock numbers and polonyna pasture use is directly related to intergenerational decline of interest and low economic competitiveness, curing cannabis as well as the time constraints on working populations . This has rippled down to demographic shifts and work migration seen Hutsulshchyna.
Migration was observed in many of the villages visited, where residents migrate seasonally to work in Poland, Russia or Western Europe with predominant sectors being seasonal agricultural work, construction, and service . Government subsidies to uphold Hutsul pastoral traditions are nonexistent in Ukraine. One recent positive development in 2020 that works to preserve bryndza, and by proxy, polonynas, is the European Union’s incorporation of bryndza as a geographical indicator. The EU states use a system of protected geographical indicators, which include names that are applied to products made within a specific area . It is the ecological processes within the landscape, climate, and soil that ensures the tradition, and its perpetuation of local economy within the region and unique taste. This is the first product in Ukraine with this geographical indication mark, ensuring its authenticity, promotion on the economic market, and guaranteeing its quality. Traditional foods in Hutsulshchyna are tethered to the landscape and the various habitats that species are found. Many berry species provide critical nutrition in the form of food, as well as medicinal, economic, and ecological importance. Bilberry and raspberry are considered the most culturally important plants. Mushrooms, such as penny bun and chanterelles, are highly sought after and a critical food source, especially during the winter religious season. Polonynas, as a critical and culturally significant habitat in Hutsulshchyna, are concretely linked to the traditional food of bryndza, as well as many other culturally important plants ; their survivals interlinked. The significance of the EU’s incorporation of bryndza as a geographical indicator provides a layer of resilience in maintaining these practices and thus ensuring food sovereignty. Attributes of socio-ecological resilience include adaptive capacity, which consists of both short-term, immediate responses and long-term, culturally valued responses . TEK, formed through ecocultural memories, is an active reflex of acknowledging rootedness to place through language, practice, and local ecologies, ultimately sustaining the adaptive capacity of Hutsul communities to survive world wars, food shortages, shifting borders, long-lasting impacts of colonialism as well as current environmental challenges.
Ecocultural memories thread together to form a dynamic knowledge base called TEK, which provides a continual opportunity for knowledge sharing within communities. It can be seen as a time-tested, repeated, readjusted knowledge base resulting in resilience. Coping strategies include gathering a diversity of foods from a diversity of habitats, mitigating the possibility of food scarcity by redistributing reliance on any one habitat type or food source. Another coping strategy includes modifying and continually adapting harvesting of where, when, and how of culturally important species are gathered, dependent on disturbances and climatic changes. Adaptive strategies include an economy of gathering which provides a diversified way of supplementing income and personal needs, while providing trade between communities. Additionally, fallback foods used in the early 20th century are still used today, with uses transforming from medicine or seasoning to food, under times of stress. Fallback foods provide a built-in coping capacity to overcome future adversities. It is the integration of coping mechanisms and adaptive strategies that provide the pathway to maintaining traditional foods in the region, which explicitly connect people to place through religious holidays, meal sharing, and customs. Food sovereignty is an emergent characteristic of community-driven, sustainably maintained ecosystems that provide culturally relevant sustenance, nurturing both community and landscape. Lab-on-a-chip systems are used to perform personalized health diagnostic tests away from the lab. Implementation of bio-assay automation reduces the cost of medical personnel and diminishes the incidence of human errors. Point-of-care diagnostic platforms are required to have low cost, low power use, be reliably automated, and free of sophisticated detection technologies. POC platforms have already been realized in the form of lateral flow immunoassays and paper fluidic colorimetric assays. Unfortunately, the aforementioned POC devices are based on capillary flow, and therefore do not work well when more complex multi-step bio-assays are performed. In these cases, some other fluid propulsion mechanisms instead of capillary flow are required.
There are many POC devices that rely on active propulsion techniques, like centrifugal propulsion, electrowetting, and magnetic beads. These tests typically entail increased fabrication and material cost, complex automation schemes, and sophisticated hardware. In this work, we developed two distinct approaches for the realization of an inexpensive automated colorimetric immunoassay with multiple wash steps: A fused deposition modeling -printed frame was used and a disposable fluidic chip that includes an elastomeric dome and fluidic channels fabricated using SLA, was printed, where the fluidic movement was facilitated by servomotors pushing on the elastomeric dome, and propelling the reagents or wash from the domed reservoirs through fluidic channels to the test chamber, or an FDM-printed non-disposable frame was used, with servomotors connected to standard inexpensive and readily-available disposable plastic syringes filled with wash and reagents to automate the steps of the assay. Draining of the test chamber was performed with a syringe attached to a servomotor, where negative pressure was created by the pulling on the syringe plunger. All automation was controlled by a program uploaded to the Arduino-based electronic board of the platform via a computer. The test results of the colorimetric bio-assay can be assessed by eye or with a smart phone for quantitative measurement, and e-mailed or texted to a hospital or doctor’s office. In this work, cannabis dryer we created a proof-of-principle platform that utilized elements of both approaches and , as outlined above. We used plastic syringes for most of the platform reagent reservoirs, including drainage and waste. The wash reservoir consisted of an SLA-printed elastomeric dome. Thus, the feasibility of both approaches was tested at the same time. The automated platform demonstrated in the presented work proved the viability of both approaches: Construction of an automated bio-assay platform using syringes only, or a printed dome-based bio-assay platform. To our knowledge, this study constitutes the first demonstration of an SLA-based dome bio-assay platform and a syringe-based automated bio-assay platform. The concepts developed in the present work build on prior research that utilized FDM to fabricate a fluidic chip with embedded microchannels. Fluid leakage was a recurring issue in the design, due to the layer-by-layer deposition nature of FDM. Our present fabrication approach utilizes syringes and photocurable resin cross linked using SLA, thus mitigating the problem of fluid leakage. The present platform employs FDM printing for the fabrication of parts that are not in contact with fluids, such as the non-disposal platform frame and the gear/rack mechanisms. Additionally, the prior fluidic platform design used molded silicone domes that needed to be sealed to the body of 3D printed fluidic chip. Our present fabrication route avoids the need to seal the domes to the plastic chip by 3D-printing the integrated dome. The proposed approach is useful in making POC bio-assays more widespread.
For example, a hospital in rural India with an FDM printer could be sent a stereolithography file to print a frame. The Arduino electronic board and other parts, such as servomotors, can be easily acquired. All the parts and materials to produce our automated platform cost less than $100, while all disposable materials, including syringes, tubing, and reagents, cost around $5 per test. If an exclusively syringe-based platform is utilized, the only disposable parts needed are plastic syringes. The syringes would be filled with a prescribed volume of reagents, and the required program uploaded to the Arduino board. This approach provides the most flexibility to perform a wide range of bio-assays on the spot, as it does not require all the reagents in typical pre-packaged volumes. This simplifies assay logistics, including that of cold storage of reagents. We expect that our proposed recipes and programs will be readily available for download by non-profit organizations and reagent makers. There is a growing interest in manufacturing functional parts inexpensively with high-aspect-ratio geometry and complex topologies. Additive manufacturing , including various types of 3D printing, is an increasingly popular form of fabrication. The AM process requires a part to be designed using a CAD program, such as SolidWorks, followed by the extrapolation of the structural information into an STL file. The STL file partitions the CAD drawing into a series of volumetric pixels, which are digitally sliced into layers along the Z direction. These layers are physically deposited onto a substrate in layer-by-layer manner, using various AM methods, including FDM and SLA. FDM and SLA accommodate a variety of material choices, including elastomeric materials, allowing for the fabrication of flexible parts such as the dome-based elastic pumps presented in this work. In this work, we conducted a brief review of microfluidic device fabrication methods, and emphasized recent developments in additive manufacturing. We followed this review with a description of the fabrication processes selected for our bio-assay platform. Lastly, we detailed an outline of our experimental procedures, and described the experimental results of a malaria bio-assay performed on our automated platform.The first traditional microfluidic devices were fabricated using glass and silicon, utilizing mostly a toolbox of lithographic techniques employed in the semiconductor industry. Equipment selection was subsequently further developed to include femtosecond lasers to fabricate glass microfluidic chips, and the selection of materials employed for fluidic chips was also expanded to include such materials as “liquid Teflon” and environmentally-sensitive materials such as hydrogels. With the advent of soft lithography and 3D printing, there was a push to develop inexpensive and reliable microfluidic platforms with bio-compatible plastics and resins, demonstrated by the recent fabrication trends outlined below.Soft lithography is based on a three-step process where microfabrication techniques are first used to produce a reusable mold, which is then filled with a curable epoxy Polydimethylsiloxane and subsequently separated from the mold after an appropriate curing time. The molded part is typically attached to a glass substrate using plasma treatment. The soft lithography approach allows for the usage of more expensive high-precision clean room lithographic techniques to produce a mold. Once the mold is produced, it can be used multiple times to fabricate identical fluidic chips, dramatically reducing the cost of individual fluidic microdevices. The soft lithography approach has allowed for much more widespread adoption of lab-on-a-chip devices, and for the production of intricate parts such as elastomeric valves. For example, the PDMS valves used by the Quake group have facilitated the implementation of large-scale microfluidic bioreactors for drug discovery and other applications. However, when the device is assembled from separately produced parts, the manufacturability is restricted, due to cumbersome alignment, bonding, and assembly. 3D printing presents an approach where a complete microfluidic device is produced without post-processing alignment and assembly steps.