Browsing by Author "Batish, Inayat"
Now showing 1 - 3 of 3
Results Per Page
Sort Options
- Developing Serum-Free Media Via Bioprocessing For Cultivated Seafood ProductsBatish, Inayat (Virginia Tech, 2022-09-08)Global food production management has become a challenge with an anticipated population of 10 billion people by 2050 and the ongoing COVID-19 epidemic. Seafood is a vital food source due to its widespread consumption, excellent nutrient profile, and low feed conversion ratio, rendering its sustainable production quintessential. Cellular agriculture or cultured meat can increase seafood production; however, the conventional use of Fetal Bovine Serum (FBS) in culture media restricts its utilization at an industrial level. FBS is effective but has many limitations: unethical animal extraction, high demand and low supply, poorly defined ingredients, variable performance, and high cost that impedes the feasibility and commercial viability of cellular agriculture. Thus, employing serum-free media becomes a quintessential need for cellular agriculture. This project aims to replace or reduce the typical 10% serum usage in Zebrafish embryonic stem cell (ESC) production media with protein hydrolysates derived from low-cost natural sources with high protein content. Enzymatic hydrolysis was performed on nine sources: insects (black army fly and cricket), plants (pea), fungi (mushroom and yeast), algae, and marine invertebrates (oyster, mussel, and lugworm). The resulting hydrolysates were evaluated for serum replacement in zebrafish ESCs. All hydrolysates were used at five different concentrations (10, 1 0.1, 0.01 and 0.001 mg/mL) in serum concentrations of 10%, 5%, and 0% with four biological replicates. The best hydrolysate sources and concentrations were selected for further testing at 2.5% and 1% serum concentrations. All hydrolysates, except for cricket, could restore or significantly increase cell growth with 50% less serum at a concentration of 0.1-0.001mg/mL. Protein hydrolysate concentration of 10 and 1mg/mL was toxic for cells. Additionally, the eight hydrolysates could reduce serum concentrations up to 75–90%. However, no protein hydrolysate could completely replace serum, as cells using only protein hydrolysates exhibited morphological aberrations and decreased growth. Replacing serum with protein hydrolysates lowers cellular agriculture's overall cost, thus enabling the commercialization of cultured meat and the development of a sustainable food system. In the future, blending various protein hydrolysate sources with or without the addition of conventional growth factors could be done to create the ideal serum-free media.
- Evaluating the Potential of Marine Invertebrate and Insect Protein Hydrolysates to Reduce Fetal Bovine Serum in Cell Culture Media for Cultivated Fish ProductionBatish, Inayat; Zarei, Mohammad; Nitin, Nitin; Ovissipour, Reza (MDPI, 2022-11-16)The use of fetal bovine serum (FBS) and the price of cell culture media are the key constraints for developing serum-free cost-effective media. This study aims to replace or reduce the typical 10% serum application in fish cell culture media by applying protein hydrolysates from insects and marine invertebrate species for the growth of Zebrafish embryonic stem cells (ESC) as the model organism. Protein hydrolysates were produced from black soldier flies (BSF), crickets, oysters, mussels, and lugworms with a high protein content, suitable functional properties, and adequate amino-acid composition, with the degree of hydrolysis from 18.24 to 33.52%. Protein hydrolysates at low concentrations from 0.001 to 0.1 mg/mL in combination with 1 and 2.5% serums significantly increased cell growth compared to the control groups (5 and 10% serums) (p < 0.05). All protein hydrolysates with concentrations of 1 and 10 mg/mL were found to be toxic to cells and significantly reduced cell growth and performance (p < 0.05). However, except for crickets, all the hydrolysates were able to restore or significantly increase cell growth and viability with 50% less serum at concentrations of 0.001, 0.01, and 0.1 mg/mL. Although cell growth was enhanced at lower concentrations of protein hydrolysates, the cell morphology was altered due to the lack of serum. The lactate dehydrogenase (LDH) activity results indicated that BSF and lugworm hydrolysates did not alter the cell membrane. In addition, light and fluorescence imaging revealed that the cell morphological features were comparable to those of the 10% serum control group. Overall, lugworm and BSF hydrolysates reduced the serum by up to 90% while preserving excellent cell health.
- Prevalence of Microplastics in the Eastern Oyster Crassostrea virginica in the Chesapeake Bay: The Impact of Different Digestion Methods on Microplastic PropertiesAung, Thet; Batish, Inayat; Ovissipour, Reza (MDPI, 2022-01-10)This study aimed to determine the microplastic prevalence in eastern oysters (C. virginica) in three sites in the Chesapeake Bay in Virginia and optimize the digestion methods. The digestion results illustrate that the lowest recovery rate and digestion recovery were related to enzymatic, enzymatic + hydrogen peroxide (H2O2), and HCl 5% treatments, while the highest digestion recovery and recovery rate were observed in H2O2 and basic (KOH) treatments. Nitric acid digestion resulted in satisfying digestion recovery (100%), while no blue polyethylene microplastics were observed due to the poor recovery rate. In addition, nitric acid altered the color, changed the Raman spectrum intensity, and melted polypropylene (PP) and polyethylene terephthalate (PET). In order to determine the number of microplastics, 144 oysters with an approximately similar size and weight from three sites, including the James River, York River, and Eastern Shore, were evaluated. Fragments were the most abundant microplastics among the different microplastics, followed by fibers and beads, in the three sites. A significantly higher number of fragments were found in the James River, probably due to the greater amount of human activities. The number of microplastics per gram of oyster tissue was higher in the James River, with 7 MPs/g tissue, than in the York River and Eastern Shore, with 6.7 and 5.6 MPs/g tissue.