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BioTech
Volume 14
Issue 4
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BioTech, Volume 14, Issue 4 (December 2025) – 24 articles

Cover Story (view full-size image): Human dental pulp stem cells (DPSCs) represent a valuable alternative to bone marrow-derived mesenchymal stem cells (MSCs) for regenerative medicine because of their easy access and minimal ethical concerns. Gene-engineered DPSCs and their derivatives [i.e., induced pluripotent stem cells (iPSCs) and differentiated progeny] are also promising candidates for future cell-based therapies, including oral regeneration. View this paper
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16 pages, 2055 KB  
Article
Enhancing Nonylphenol Biodegradation: The Role of Acetyl-CoA C-Acetyltransferase in Bacillus cereus
by Fanglian Lu, Deqin Luo, Lian Yang and Ranran Dong
BioTech 2025, 14(4), 99; https://doi.org/10.3390/biotech14040099 - 18 Dec 2025
Viewed by 309
Abstract
Nonylphenol (NP) bioremediation is constrained by the scarcity of efficient and non-pathogenic degrading strains. To clarify the role of acetyl-CoA C-acetyltransferase (AtoB) in NP degradation, we generated an atoB-overexpressed strain (LY-OE) from the environmentally tolerant Bacillus cereus LY and compared its degradation [...] Read more.
Nonylphenol (NP) bioremediation is constrained by the scarcity of efficient and non-pathogenic degrading strains. To clarify the role of acetyl-CoA C-acetyltransferase (AtoB) in NP degradation, we generated an atoB-overexpressed strain (LY-OE) from the environmentally tolerant Bacillus cereus LY and compared its degradation rate with the wild type using HPLC. Untargeted lipidomics was conducted to characterize metabolic responses under NP stress, and key differential lipid metabolites (DELMs) were further validated by ELISA. Additionally, AtoB concentration and ATP content were quantified using commercial assay kits in Bacillus cereus. LY-OE showed a markedly higher NP degradation rate (96%) than LY (85%). Lipidomic analysis identified 34 significant DELMs (VIP > 1, p < 0.05), including elevated cardiolipin (CL) and phosphatidylglycerol (PG), and reduced phosphatidylcholine (PC) and triglycerides (TG). ELISA confirmed these changes (p < 0.01 or p < 0.001), consistent with lipidomic findings. LY-OE showed significantly higher AtoB concentration during the logarithmic growth phase and exhibited higher ATP content during NP degradation. These findings suggest that atoB overexpression enhances NP degradation by both boosting energy supply and remodeling lipid metabolism. This work identifies atoB as a key factor for NP biodegradation and provides a promising strategy for developing high-performance bioremediation strains. Full article
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20 pages, 1197 KB  
Review
Ion Mobility–Mass Spectrometry Imaging: Advances in Biomedical Research
by Mengya Liu, Chi Zhang, Lili Xu, Md. Muedur Rahman, Shoshiro Hirayama, Shuhei Aramaki, Atsushi Baba, Ryo Omagari, Yutaka Takahashi, Tomoaki Kahyo and Mitsutoshi Setou
BioTech 2025, 14(4), 98; https://doi.org/10.3390/biotech14040098 - 18 Dec 2025
Viewed by 897
Abstract
Mass spectrometry imaging (MSI) visualizes the spatial distribution of biomolecules in tissues, whereas ion mobility–mass spectrometry (IM-MS) separates ions through the collision cross-section (CCS) with an inert gas, providing the structural characteristics of isomers. Recent advances have established an integrated workflow, ion mobility–mass [...] Read more.
Mass spectrometry imaging (MSI) visualizes the spatial distribution of biomolecules in tissues, whereas ion mobility–mass spectrometry (IM-MS) separates ions through the collision cross-section (CCS) with an inert gas, providing the structural characteristics of isomers. Recent advances have established an integrated workflow, ion mobility–mass spectrometry imaging (IM-MSI), that couples IM with MSI, uniting molecular discrimination with spatial mapping. This synergy has been widely applied in oncology and neuropsychiatric disorders, offering unprecedented insights into biomarker discovery and disease mechanisms. Here, we summarize the principles and classifications of IM-MSI, review their combined biomedical applications, and discuss data processing workflows and commonly used tools. Full article
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21 pages, 6736 KB  
Review
From Traditional Use to Molecular Mechanisms: A Bioinformatic and Pharmacological Review of the Genus Kalanchoe with In Silico Evidence
by Cristián Raziel Delgado-González, Ashutosh Sharma, Margarita Islas-Pelcastre, Mariana Saucedo-García, Eliazar Aquino-Torres, Jaime Pacheco-Trejo, Silvia Armenta-Jaime, Nallely Rivero-Pérez and Alfredo Madariaga-Navarrete
BioTech 2025, 14(4), 97; https://doi.org/10.3390/biotech14040097 - 12 Dec 2025
Viewed by 664
Abstract
The genus Kalanchoe (Crassulaceae) comprises approximately 125 species of succulents distributed across Madagascar, Africa, Arabia, Australia, Southeast Asia, and tropical America. Traditionally regarded as “miracle plants”, Kalanchoe species are employed for treating inflammatory, infectious, metabolic, and cardiovascular conditions; this is associated with their [...] Read more.
The genus Kalanchoe (Crassulaceae) comprises approximately 125 species of succulents distributed across Madagascar, Africa, Arabia, Australia, Southeast Asia, and tropical America. Traditionally regarded as “miracle plants”, Kalanchoe species are employed for treating inflammatory, infectious, metabolic, and cardiovascular conditions; this is associated with their abundant content of polyphenols, including phenolic acids and flavonoids such as quercetin, kaempferol, luteolin, rutin, and patuletin. However, robust clinical evidence remains limited. This review integrates pharmacological and bioinformatic perspectives by analyzing more than 70 studies published since 2000 on 15 species, including Bryophyllum. As an in silico complement, the genome of Kalanchoe fedtschenkoi was used to predict genes (AUGUSTUS), perform homology searches against Arabidopsis thaliana, and model three key enzymes: CHS, CYP90, and VEP1. The AlphaFold2/ColabFold models showed conserved catalytic motifs, and molecular docking with representative ligands supported the plausibility of biosynthetic pathways for flavonoids, brassinosteroids, and bufadienolides. The available evidence highlights chemopreventive, antibacterial, anti-inflammatory, antiviral, antioxidant, and cytotoxic activities, primarily associated with flavonoids and bufadienolides. Significant gaps remain, such as the lack of gene–metabolite correlations and the absence of standardized clinical trials. Overall, Kalanchoe represents a promising model that requires multi-omics approaches to enhance its phytopharmaceutical potential. Full article
(This article belongs to the Topic From Plant to Pharmacology: Understanding the Metabolism of Natural Products)
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17 pages, 1048 KB  
Article
Lowly Expressed Toxin Transcripts in Poorly Characterized Myanmar Russell’s Viper Venom Gland
by Khin Than Yee, Jason Macrander, Olga Vasieva and Ponlapat Rojnuckarin
BioTech 2025, 14(4), 96; https://doi.org/10.3390/biotech14040096 - 4 Dec 2025
Viewed by 519
Abstract
In Myanmar, Russell’s viper (Daboia siamensis) bite is a significant public health problem. In this study, we expend upon our previous RNA-sequencing approach to characterize candidate toxin genes encoding D. siamensis toxins. The mRNA was extracted from Myanmar Russell’s viper venom [...] Read more.
In Myanmar, Russell’s viper (Daboia siamensis) bite is a significant public health problem. In this study, we expend upon our previous RNA-sequencing approach to characterize candidate toxin genes encoding D. siamensis toxins. The mRNA was extracted from Myanmar Russell’s viper venom glands. The RNAseq was performed using Illumina next-generation sequencing. Subsequently, candidate toxin transcripts were recognized by the Venomix pipeline. This study focused on 29 unique cDNA sequences representing eight newly identified venom gene families with low-to-moderate expression levels. These transcripts represented 0.088% of the total number of transcripts in the dataset. The translated protein sequences were analyzed for their conserved motifs and domains to predict their functions. They were neprilysins (bioactive peptide inactivators), cystatins (protease inhibitors with anti-metastatic activities), waprin and vipericidin (antimicrobial peptides), veficolin (platelet and complement activation), vespryns and three-finger toxins (elapid toxin homologs causing neurotoxic activity and tissue damage), and endothelial lipases (unknown function). Their functional activities should be further investigated for potential therapeutic applications, for example, in cancer or antibiotic-resistant infections. Full article
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19 pages, 3800 KB  
Article
Maize miRNAs Might Regulate Human Genes Involved in Prostate Cancer: An In Silico Approach
by Ximena Vázquez-Cadena, Oscar Alejandro Faz-Cortez, Benito Pereyra-Alférez, César Ignacio Hernández-Vásquez, Luis Jesús Galán-Wong, Myriam Elías-Santos and Jorge Hugo Garcia-Garcia
BioTech 2025, 14(4), 95; https://doi.org/10.3390/biotech14040095 - 3 Dec 2025
Viewed by 624
Abstract
MicroRNAs (miRNAs) are small single-stranded non-coding RNA molecules that regulate gene expression at the post-transcriptional level. Recent studies have demonstrated that plant miRNAs can survive through dietary intake and act as signaling molecules in intercellular communication, proving a cross-kingdom interaction. The aim of [...] Read more.
MicroRNAs (miRNAs) are small single-stranded non-coding RNA molecules that regulate gene expression at the post-transcriptional level. Recent studies have demonstrated that plant miRNAs can survive through dietary intake and act as signaling molecules in intercellular communication, proving a cross-kingdom interaction. The aim of the present study was to use computational approaches to identify interactions between Zea mays (maize) miRNAs and human coding mRNAs potentially involved in different biological processes. We identified 961 unique genes potentially regulated by maize miRNAs. Furthermore, functional enrichment analysis via GO and KEGG was carried out focusing primarily on the pathway related to prostate cancer where 13 genes were potentially regulated by 15 maize miRNAs. Our findings not only provide an important insight into the potential effects that maize-derived miRNAs could have on the human body, but also highlight the importance of considering these molecules for further research and potential therapeutic applications against major diseases such as cancer. Full article
(This article belongs to the Section Computational Biology)
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23 pages, 3391 KB  
Review
A Comprehensive Review of Plant and Microbial Natural Compounds as Sources of Potential Helicobacter pylori-Inhibiting Agents
by Srichandan Padhi, Swati Sharma, Puja Sarkar, Marco Masi, Alessio Cimmino and Amit Kumar Rai
BioTech 2025, 14(4), 94; https://doi.org/10.3390/biotech14040094 - 26 Nov 2025
Viewed by 1105
Abstract
Helicobacter pylori, the gastric pathogen which colonizes the gastric mucosa of more than half of the world’s population, is considered a risk factor for peptic ulcers and is epidemiologically associated with gastric cancer. Antimicrobial eradication of this pathogen has now become a [...] Read more.
Helicobacter pylori, the gastric pathogen which colonizes the gastric mucosa of more than half of the world’s population, is considered a risk factor for peptic ulcers and is epidemiologically associated with gastric cancer. Antimicrobial eradication of this pathogen has now become a central concern because of its growing resistance to frontline antibiotics such as clarithromycin and metronidazole. Moreover, these antibiotics can have adverse effects on the normal human gut flora and can lead to several health complications. Most times, the antibiotic doses become intolerable to the elderly population and they reject the therapy. This has impelled us to think about alternate effective and safe antimicrobials which can replace antibiotic usage or may reduce their dosage when used together with the antibiotics. Plant and microbial natural products, in view of this, offer an excellent source of novel and potential antimicrobial agents. Herein, we review anti-H. pylori natural compounds from diverse plant and microbial sources and highlight their role in the management of H. pylori infection. Full article
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23 pages, 1788 KB  
Review
Somatic Embryogenesis: A Biotechnological Route in the Production of Recombinant Proteins
by Marco A. Ramírez-Mosqueda, Jorge David Cadena-Zamudio, Carlos A. Cruz-Cruz, José Luis Aguirre-Noyola, Raúl Barbón, Rafael Gómez-Kosky and Carlos Angulo
BioTech 2025, 14(4), 93; https://doi.org/10.3390/biotech14040093 - 26 Nov 2025
Cited by 2 | Viewed by 851
Abstract
Somatic embryogenesis (SE) is a morphogenetic pathway widely employed in the commercial micropropagation of plants. This route enables the generation of somatic embryos from somatic tissues, which give rise to complete (bipolar) plants that develop like zygotic embryos. SE can proceed via direct [...] Read more.
Somatic embryogenesis (SE) is a morphogenetic pathway widely employed in the commercial micropropagation of plants. This route enables the generation of somatic embryos from somatic tissues, which give rise to complete (bipolar) plants that develop like zygotic embryos. SE can proceed via direct or indirect pathways, and both approaches have been adapted not only for large-scale clonal propagation but also for the regeneration of genetically modified plants. In this context, SE can be harnessed as a versatile platform for recombinant protein production, including vaccine antigens and therapeutic proteins, by combining plant tissue culture with genetic transformation strategies. Successful examples include non-model plants, as Daucus carota and Eleutherococcus senticosus expressing the cholera and heat-labile enterotoxin B subunits, respectively; Oryza sativa, Nicotiana tabacum, and Medicago sativa producing complex proteins such as human serum albumin (HSA), α1-antitrypsin (AAT), and monoclonal antibodies. However, challenges remain in optimizing transformation efficiency, scaling up bioreactor-based suspension cultures, and ensuring proper post-translational modifications under Good Manufacturing Practice (GMP) standards. Recent advances in synthetic biology, modular vector design, and glycoengineering have begun to address these limitations, improving control over transcriptional regulation and protein quality. This review highlights the application of SE as a biotechnological route for recombinant protein production, discusses current challenges, and presents innovative strategies and perspectives for the development of sustainable plant-derived biopharmaceutical systems. Full article
(This article belongs to the Special Issue Plant Biotechnology in the Fight Against Human Diseases)
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18 pages, 3118 KB  
Article
Synergistic Effect of Bacillus subtilis B3 and β-Glucanase on Solid-State Fermentation of Sunflower Meal
by Shuqi Chen, Haoran Shi, Peng Zhao, Zengqiang Ma, Xiaolong Li, Xiangyu Wang and Feiyan Xue
BioTech 2025, 14(4), 92; https://doi.org/10.3390/biotech14040092 - 18 Nov 2025
Viewed by 556
Abstract
Background: Sunflower meal (SFM), a promising feed material, is constrained by its high content of crude fiber (CF) and chlorogenic acid (CGA). Methods: This study utilized a synergistic solid-state fermentation process involving the Bacillus subtilis strain B3 and the enzyme β-glucanase to enhance [...] Read more.
Background: Sunflower meal (SFM), a promising feed material, is constrained by its high content of crude fiber (CF) and chlorogenic acid (CGA). Methods: This study utilized a synergistic solid-state fermentation process involving the Bacillus subtilis strain B3 and the enzyme β-glucanase to enhance SFM’s application potential. Results: The synergistic treatment notably reduced CF by 12.7% and CGA by 99.77%, while simultaneously increasing acid-soluble protein and reducing sugar by 111.3% and 283.1%, respectively. Positive impacts on its physical structure, characterized by a looser network with visible pores, and on its microbial community, evidenced by an enriched abundance of fungal species such as Cyberlindnera and Aspergillus, were also observed. In vitro assays indicated improved digestibility of dry matter, neutral detergent fiber, and crude protein, along with a non-significant reduction in methane production. Conclusions: These results demonstrate that microbial-enzymatic synergy effectively enhances SFM’s nutritional profile. Full article
(This article belongs to the Section Industry, Agriculture and Food Biotechnology)
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9 pages, 672 KB  
Communication
Concurrent Analysis of Antioxidant and Pro-Oxidant Activities in Compounds from Plant Cell Cultures
by Marcela Blažková, Ľubica Uváčková, Mária Maliarová, Jozef Sokol, Jana Viskupičová and Tibor Maliar
BioTech 2025, 14(4), 91; https://doi.org/10.3390/biotech14040091 - 14 Nov 2025
Viewed by 593
Abstract
Oxidative stress reflects an imbalance between pro-oxidants and antioxidants arising from physiological or environmental factors. Here, we applied our previously developed in situ microplate method for the simultaneous determination of antioxidant and pro-oxidant activities to compounds produced by plant cell cultures in vitro. [...] Read more.
Oxidative stress reflects an imbalance between pro-oxidants and antioxidants arising from physiological or environmental factors. Here, we applied our previously developed in situ microplate method for the simultaneous determination of antioxidant and pro-oxidant activities to compounds produced by plant cell cultures in vitro. The primary aim was to evaluate the added value of these compounds, which are widely used as additives in food, cosmetic, and pharmaceutical products. The secondary aim was to assess whether a predominance of pro-oxidant activity could limit their biotechnological production. Thirty-three compounds known to be produced by in vitro cultures (polyphenolic acids, flavonoids, quinones, alkaloids, etc.) were tested, and the pro-oxidant–antioxidant balance index (PABI) was calculated. Sixteen compounds showed measurable activities with DPPH50/FRAP50 values below 2 mM. Within this set, rosmarinic acid exhibited pronounced pro-oxidant behavior, whereas gallic acid, chlorogenic acid, and the anthocyanin cyanidin showed higher antioxidant potency and favorable PABI values. Such compounds may deliver added benefits when incorporated into food or cosmetic products and are unlikely to limit production in cell culture. Full article
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26 pages, 4642 KB  
Article
Acylpyruvates and Their Heterocyclic Derivatives as Growth Regulators in Chlorella vulgaris
by Anastasia D. Novokshonova, Pavel V. Khramtsov, Maksim V. Dmitriev and Ekaterina E. Khramtsova
BioTech 2025, 14(4), 90; https://doi.org/10.3390/biotech14040090 - 10 Nov 2025
Viewed by 796
Abstract
Acylpyruvate derivatives represent a promising yet underexplored class of compounds for modulating microalgal growth and metabolism. Inspired by the metabolic role of pyruvate and the diverse bioactivity of its acylated analogs, this study investigates the structure–activity relationship of a diverse library of 55 [...] Read more.
Acylpyruvate derivatives represent a promising yet underexplored class of compounds for modulating microalgal growth and metabolism. Inspired by the metabolic role of pyruvate and the diverse bioactivity of its acylated analogs, this study investigates the structure–activity relationship of a diverse library of 55 acylpyruvate-derived compounds for stimulation of the green microalga Chlorella vulgaris. The library, encompassing 12 chemotypes including acylpyruvic acids, their esters, and various heterocyclic derivatives, was screened for effects on C. vulgaris growth. Six compounds were identified as active ones that enhanced biomass production in a preliminary microassay. Notably, four of these active compounds were direct acylpyruvate derivatives, highlighting this scaffold as the most promising one. Conversely, a specific subclass, 1,4-benzoxazin-2-ones, exhibited potent, dose-dependent algicidal activity. Detailed assessment of the active compounds under scaled-up culture conditions revealed that while their effect on overall cell density was limited, several compounds significantly enhanced the intracellular content of valuable metabolites: one increased chlorophyll content by 17%, another elevated carotenoids by 40%, and a third boosted neutral lipid accumulation by 44%. Furthermore, control experiments confirmed that the bioactivity of p-ethoxybenzoylpyruvates, which showed the best biological activity, is inherent in the intact framework and is not mediated by their hydrolysis products. Our findings underscore the potential of acylpyruvates as versatile tools for the enhancement of metabolite production in microalgae and as potent candidates for the development of algicides. Full article
(This article belongs to the Special Issue A Sustainable Approach to Macroalgae and Microalgae: From Cultivation to Compound Recovery and Biotechnological Applications)
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35 pages, 1059 KB  
Review
Improving the Antioxidant Potential of Berry Crops Through Genomic Advances and Modern Agronomic and Breeding Tools
by Georgios Mitronikas, Athina Voudanta, Aliki Kapazoglou, Maria Gerakari, Eleni M. Abraham, Eleni Tani and Vasileios Papasotiropoulos
BioTech 2025, 14(4), 89; https://doi.org/10.3390/biotech14040089 - 7 Nov 2025
Viewed by 1029
Abstract
The growing demand for sustainable, health-promoting foods has intensified efforts to improve the antioxidant potential of berry crops through integrative agronomic, genomic, and breeding innovations. Berries are rich dietary sources of bioactive compounds that support human health and provide benefits far beyond basic [...] Read more.
The growing demand for sustainable, health-promoting foods has intensified efforts to improve the antioxidant potential of berry crops through integrative agronomic, genomic, and breeding innovations. Berries are rich dietary sources of bioactive compounds that support human health and provide benefits far beyond basic nutrition. This review explores the diversity of major berry crops, including blueberries, raspberries, cranberries, blackberries, and grapes, with emphasis on their nutritional value and antioxidant profiles. It also examines their domestication history, wild relatives, and commercial cultivars, offering insight into the genetic and phenotypic diversity underlying their rich chemical composition. Furthermore, the review highlights the application of modern tools to enhance antioxidant content. By integrating agronomic practices such as seed priming and grafting, advanced molecular breeding technologies, including multi-omics, genome-wide association studies (GWAS), and genome editing, breeders and researchers can accelerate the development of high-value berry cultivars that combine superior nutritional quality, resilience to environmental stress, and sustainable productivity under the challenges posed by climate change. Full article
(This article belongs to the Section Industry, Agriculture and Food Biotechnology)
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46 pages, 1109 KB  
Review
Engineered Human Dental Pulp Stem Cells with Promising Potential for Regenerative Medicine
by Emi Inada, Issei Saitoh, Masahiko Terajima, Yuki Kiyokawa, Naoko Kubota, Haruyoshi Yamaza, Kazunori Morohoshi, Shingo Nakamura and Masahiro Sato
BioTech 2025, 14(4), 88; https://doi.org/10.3390/biotech14040088 - 3 Nov 2025
Cited by 1 | Viewed by 2119
Abstract
The fields of regenerative medicine and stem cell-based tissue engineering hold great potential for treating a wide range of tissue and organ defects. Stem cells are ideal candidates for regenerative medicine because they are undifferentiated cells with the capacity for self-renewal, rapid proliferation, [...] Read more.
The fields of regenerative medicine and stem cell-based tissue engineering hold great potential for treating a wide range of tissue and organ defects. Stem cells are ideal candidates for regenerative medicine because they are undifferentiated cells with the capacity for self-renewal, rapid proliferation, multilineage differentiation, and expression of pluripotency-associated genes. Human dental pulp stem cells (DPSCs) consist of various cell types (including stem cells) and possess multilineage differentiation potential. Owing to their easy isolation and rapid proliferation, DPSCs and their derivatives are considered promising candidates for repairing injured tissues. Recent advances in gene engineering have enabled cells to express specific genes of interest, leading to the secretion of medically important proteins or the alteration of cell behavior. For example, transient expression of Yamanaka’s factors in DPSCs can induce transdifferentiation into induced pluripotent stem cells (iPSCs). These gene-engineered cells represent valuable candidates for regenerative medicine, including stem cell therapies and tissue engineering. However, challenges remain in their development and application, particularly regarding safety, efficacy, and scalability. This review summarizes current knowledge on gene-engineered DPSCs and their derivatives and explores possible clinical applications, with a special focus on oral regeneration. Full article
(This article belongs to the Section Medical Biotechnology)
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16 pages, 2708 KB  
Article
Comparing Handcrafted Radiomics Versus Latent Deep Learning Features of Admission Head CT for Hemorrhagic Stroke Outcome Prediction
by Anh T. Tran, Junhao Wen, Gaby Abou Karam, Dorin Zeevi, Adnan I. Qureshi, Ajay Malhotra, Shahram Majidi, Niloufar Valizadeh, Santosh B. Murthy, Mert R. Sabuncu, David Roh, Guido J. Falcone, Kevin N. Sheth and Seyedmehdi Payabvash
BioTech 2025, 14(4), 87; https://doi.org/10.3390/biotech14040087 - 2 Nov 2025
Viewed by 1179
Abstract
Handcrafted radiomics use predefined formulas to extract quantitative features from medical images, whereas deep neural networks learn de novo features through iterative training. We compared these approaches for predicting 3-month outcomes and hematoma expansion from admission non-contrast head CT in acute intracerebral hemorrhage [...] Read more.
Handcrafted radiomics use predefined formulas to extract quantitative features from medical images, whereas deep neural networks learn de novo features through iterative training. We compared these approaches for predicting 3-month outcomes and hematoma expansion from admission non-contrast head CT in acute intracerebral hemorrhage (ICH). Training and cross-validation were performed using a multicenter trial cohort (n = 866), with external validation on a single-center dataset (n = 645). We trained multiscale U-shaped segmentation models for hematoma segmentation and extracted (i) radiomics from the segmented lesions and (ii) two latent deep feature sets—from the segmentation encoder and a generative autoencoder trained on dilated lesion patches. Features were reduced with unsupervised Non-Negative Matrix Factorization (NMF) to 128 per set and used—alone or in combination—for six machine-learning classifiers to predict 3-month clinical outcomes and (>3, >6, >9 mL) hematoma expansion thresholds. The addition of latent deep features to radiomics numerically increased model prediction performance for 3-month outcomes and hematoma expansion using Random Forest, XGBoost, Extra Trees, or Elastic Net classifiers; however, the improved accuracy only reached statistical significance in predicting >3 mL hematoma expansion. Clinically, these consistent but modest increases in prediction performance may improve risk stratification at the individual level. Nevertheless, the latent deep features show potential for extracting additional clinically relevant information from admission head CT for prognostication in hemorrhagic stroke. Full article
(This article belongs to the Special Issue Advances in Bioimaging Technology)
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13 pages, 1451 KB  
Article
Exposure to Sulfur Hexafluoride Influences Viability in Cell Transplant Suspensions
by Laura Martínez-Alarcón, Sergio Liarte, Juana M. Abellaneda, Juan J. Quereda, Livia Mendonça, Antonio Muñoz, Pablo Ramírez and Guillermo Ramis
BioTech 2025, 14(4), 86; https://doi.org/10.3390/biotech14040086 - 31 Oct 2025
Viewed by 623
Abstract
Cell transplantation is often performed with ultrasonographic guidance for accurate delivery through injection. In such procedures, using ultrasonographic contrast greatly improves target delivery. However, accumulating evidence suggests that exposure to such contrast agents may have negative effects on transplanted cells. No study so [...] Read more.
Cell transplantation is often performed with ultrasonographic guidance for accurate delivery through injection. In such procedures, using ultrasonographic contrast greatly improves target delivery. However, accumulating evidence suggests that exposure to such contrast agents may have negative effects on transplanted cells. No study so far has researched this issue. Stabilized sulfur hexafluoride (SF6) microbubbles are a widely used sonographic contrast agent. Skin hCD55 porcine transgenic fibroblasts and mesenchymal stem cells from human bone marrow (hMSCs) were exposed in vitro to SF6 in concentrations ranging from 1.54 µM to 308 µM. The effects on viability and cell growth were registered using an impedance-based label-free Real-Time Cell Analyzer (RTCA). Data was recorded every 15 min for 50 h of total study time. Both cell lines behave distinctly when exposed to SF6. Porcine fibroblast growth showed relevant alterations only when exposed to higher concentrations. In contrast, hMSCs showed progressive growth decrease in relation to SF6 concentration. Taken together, while SF6-based contrast agents pose no threat to patient safety, our results indicate that exposure of suspended stem cells to the contrast agent could affect the effective dose administered in cell therapy procedures. This prompts specific cell lineage testing, adjusting methods and properly compensating for cell loss, with a potential impact on procedural cost and success rates. Full article
(This article belongs to the Section Medical Biotechnology)
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12 pages, 2048 KB  
Article
Recombinant Clostridium acetobutylicum Endoxylanase for Xylooligosaccharide Production from Pretreated Lignocellulosic Biomass
by Afifa Husna, Agustin Krisna Wardani, Chun-Yi Hu and Yo-Chia Chen
BioTech 2025, 14(4), 85; https://doi.org/10.3390/biotech14040085 - 30 Oct 2025
Viewed by 699
Abstract
Xylooligosaccharides (XOS) are functional oligosaccharides with recognized prebiotic properties and growing industrial relevance, typically obtained through enzymatic depolymerization of xylan-rich lignocellulosic substrates. In this study, a recombinant endo-β-1,4-xylanase (XynA) from Clostridium acetobutylicum was employed for XOS production. The xynA gene was cloned into [...] Read more.
Xylooligosaccharides (XOS) are functional oligosaccharides with recognized prebiotic properties and growing industrial relevance, typically obtained through enzymatic depolymerization of xylan-rich lignocellulosic substrates. In this study, a recombinant endo-β-1,4-xylanase (XynA) from Clostridium acetobutylicum was employed for XOS production. The xynA gene was cloned into the expression vector pET-21a(+) and heterologously expressed in Escherichia coli BL21(DE3) under induction with isopropyl β-D-1-thiogalactopyranoside (IPTG). The recombinant protein, with an estimated molecular mass of 37.5 kDa, was verified by SDS-PAGE and Western blot analysis. Functional characterization via thin-layer chromatography revealed that XynA efficiently hydrolyzed beechwood xylan and rye arabinoxylan, predominantly yielding xylobiose. Additionally, the enzyme catalyzed the conversion of xylotriose into xylobiose and trace amounts of xylose. Notably, XynA demonstrated hydrolytic activity against autohydrolysed and alkali-pretreated coconut husk biomass, facilitating the release of XOS. These results underscore the potential of C. acetobutylicum XynA as a biocatalyst for the valorization of lignocellulosic residues into high-value oligosaccharides. Full article
(This article belongs to the Special Issue BioTech: 5th Anniversary)
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12 pages, 827 KB  
Communication
Enhanced Succinate Production in Actinobacillus succinogenes via Neutral Red Bypass Reduction in a Novel Bioelectrochemical System
by Julian Tix, Fernando Pedraza, Roland Ulber and Nils Tippkötter
BioTech 2025, 14(4), 84; https://doi.org/10.3390/biotech14040084 - 29 Oct 2025
Viewed by 770
Abstract
Carbon capture and power-to-X are becoming increasingly relevant in the context of decarbonization and supply security. Actinobacillus succinogenes is capable of transforming CO2 into succinate, whereby product formation is significantly limited by the availability of NADH. The aim of this work was [...] Read more.
Carbon capture and power-to-X are becoming increasingly relevant in the context of decarbonization and supply security. Actinobacillus succinogenes is capable of transforming CO2 into succinate, whereby product formation is significantly limited by the availability of NADH. The aim of this work was to further develop a bioelectrochemical system (BES) in order to provide additional reduction equivalents and thus increase yield and titer. To this end, a new BES configuration was established. A conventional stirred tank reactor (STR) is coupled via a bypass to an H-cell, in which the redox mediator neutral red (NR) is electrochemically reduced and then returned back to the bioreactor. The indirect electron transfer decouples the electrochemical reduction from the biology and results in increased intracellular availability of NADH. The present approach resulted in an increase in yield from 0.64 g·g−1 to 0.76 g·g−1, corresponding to an increase of 18%. At the same time, a titer of 16.48 ± 0.19 g·L−1 was achieved in the BES, compared to 12.05 ± 0.18 g·L−1 in the control. The results show that the mediator-assisted, partially decoupled BES architecture significantly improves CO2-based succinate production and opens up a scalable path to the use of renewable electricity as a reduction source in power-to-X processes. Full article
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15 pages, 3966 KB  
Article
Preparation of Suaeda Tea Through Semi-Solid Fermentation Utilizing Kluyveromyces marxianus, Komagataeibacter europaeus, and Acetobacter schutzenbachii: Physicochemical Characteristics, Process Optimization, and Antioxidant Activity
by Aoqi Dong, Xiaoying Dong, Xinying Dai, Yanru Gao, Yuewen Ning, Xiya Fan and Haiyan Liu
BioTech 2025, 14(4), 83; https://doi.org/10.3390/biotech14040083 - 28 Oct 2025
Viewed by 471
Abstract
Suaeda salsa, an annual herb belonging to the genus Suaeda within the Chenopodiaceae family, is highly salt-tolerant and can thrive in large quantities on saline and alkaline soils. This study presents a novel fermentation technique to produce Suaeda tea, utilizing a synergistic [...] Read more.
Suaeda salsa, an annual herb belonging to the genus Suaeda within the Chenopodiaceae family, is highly salt-tolerant and can thrive in large quantities on saline and alkaline soils. This study presents a novel fermentation technique to produce Suaeda tea, utilizing a synergistic blend of microbial agents: Kluyveromyces marxianus, Komagataeibacter europaeus, and Acetobacter schutzenbachii. The resulting tea demonstrates a potent antioxidant capacity, with a hydroxyl radical scavenging rate of 64.2% and an exceptional 1,1-diphenyl-2-picrylhydrazyl radical scavenging capacity of 83.3%, along with increased ferric ion reduction/antioxidant power (FRAP) reducing power (1.82), indicating its superior antioxidant profile. Through the comparison of different microbial strain combinations under varying process parameters such as fermentation temperature and duration, the experiment revealed that fermentation at 37 °C for 24 h results in the highest concentrations of tea polyphenols (TPs) (≥10.87 mg/mL) and free amino acids (26.32 mg/100 mL). The quality of the fermented Suaeda tea meets the stringent GB/T 21733-2008 standards for tea beverages, exhibiting excellent physicochemical indices and sensory attributes. The antioxidant efficacy of the fermented Suaeda tea persists significantly throughout a 180-day duration. The optimization of the fermentation process for Suaeda tea not only provides a theoretical framework for large-scale production but also establishes a foundation for Suaeda salsa in the tea beverage sector. This innovation enriches the market with a diverse range of health-promoting teas, catering to the growing consumer demand for nutritious and beneficial beverages. Full article
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23 pages, 3638 KB  
Article
Extracts of Argemone mexicana L. Contain Antifungal Compounds for the In Vitro Control of Monilinia fructicola, Colletotrichum gloeosporioides, Fusarium oxysporum, and Sclerotinia sclerotiorum: Preliminary Evidence for Field Application
by Iridiam Hernández-Soto, Antonio Juárez-Maldonado, Alfredo Madariaga-Navarrete, Ashutosh Sharma, Antonio de Jesus Cenobio-Galindo, Jose Manuel Pinedo-Espinoza, Aracely Hernández-Pérez and Alma Delia Hernández-Fuentes
BioTech 2025, 14(4), 82; https://doi.org/10.3390/biotech14040082 - 26 Oct 2025
Viewed by 1548
Abstract
Argemone mexicana L. is considered a weed; however, it contains secondary metabolites that can control phytopathogenic fungi in vitro, with the potential to adapt its effectiveness in the field. In the present study, leaf extracts of A. mexicana (hexane and methanol) were prepared, [...] Read more.
Argemone mexicana L. is considered a weed; however, it contains secondary metabolites that can control phytopathogenic fungi in vitro, with the potential to adapt its effectiveness in the field. In the present study, leaf extracts of A. mexicana (hexane and methanol) were prepared, and their chemical profiles were analyzed using gas chromatography–mass spectrometry (GC-MS). The in vitro antifungal activity of each extract was evaluated at different concentrations (500, 1000, 2000, 4000, and 8000 mg L−1) against phytopathogens such as Monilinia fructicola, Colletotrichum gloeosporioides, Fusarium oxysporum, and Sclerotinia sclerotiorum. Based on their chemical profiles, 14 compounds were identified in the hexanic extract, and 11 compounds were identified in the methanolic extract. These compounds included those with antifungal activity, such as Benzene; 1.3-bis(1.1-dimethylethyl)-; pentanoic acid; 5-hydroxy-, 2,4-di-1-butylphenyl esters; 1,2,4-Triazol-4-amine; and N-(2-thienylmethyl). The hexanic extract demonstrated fungistatic activity on the four fungi tested, while the methanolic extract exhibited fungicidal activity against C. gloeosporioides and F. oxysporum. The results of the Probit analysis showed variations in the sensitivity of phytopathogenic fungi to the treatments evaluated. In M. fructicola, the hexane extract presented an EC50 of 317,146 mg L−1 and an EC90 of 400,796 mg L−1. For C. gloeosporioides, the EC50 was 2676 mg L−1 and the EC90 was 888,177 mg L−1, while in F. oxysporum an EC50 of 34,274 mg L−1 and an EC90 of 1528 mg L−1 were estimated. In the case of S. sclerotiorum, an EC50 of 560 mg L−1 and an EC90 of 7776 mg L−1 were obtained. Finally, for the commercial fungicide Captan®, an EC50 of 1.19 mg L−1 and an EC90 of 1.67 mg L−1. These results suggest that extracts from A. mexicana could provide a natural alternative for the control of phytopathogenic fungi. Full article
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11 pages, 6634 KB  
Communication
A Simple and Safe Protocol for Intra-Testicular Gene Delivery in Neonatal Mice Using a Convenient Isoflurane-Based Anesthesia System
by Kazunori Morohoshi, Miho Ohba, Masahiro Sato and Shingo Nakamura
BioTech 2025, 14(4), 81; https://doi.org/10.3390/biotech14040081 - 22 Oct 2025
Viewed by 726
Abstract
Newborn mice (up to 6 d after birth) are suitable for genetic manipulations, such as facial vein-mediated injection, owing to their hairless and thin skin. Their small body volumes also facilitate the rapid dissemination of injected solutions, supporting gene engineering-related experiments. However, anesthesia [...] Read more.
Newborn mice (up to 6 d after birth) are suitable for genetic manipulations, such as facial vein-mediated injection, owing to their hairless and thin skin. Their small body volumes also facilitate the rapid dissemination of injected solutions, supporting gene engineering-related experiments. However, anesthesia in newborns is challenging because of the potential risks associated with anesthetic agents. Isoflurane inhalation anesthesia is an option, although its effects on brain development remain under investigation. In this study, we established a reproducible protocol for delivering nucleic acids to juvenile mouse testes using a simple isoflurane-based anesthetic system prepared from common laboratory equipment. Using this system, nucleic acids were successfully delivered to juvenile mouse testes via intra-testicular injection, followed by in vivo electroporation. The present isoflurane-based method achieved >90% postoperative survival with normal maternal nursing observations. Gene delivery resulted in limited transfection of seminiferous tubules but efficient interstitial Leydig cell transfection. Thus, gene engineering in somatic and germ cells in neonatal mice will be facilitated using the anesthetic protocol established in this study. Full article
(This article belongs to the Section Biotechnology Regulation)
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25 pages, 5782 KB  
Review
Molecular Docking as a Key Driver of Biocontrol for Agri-Food Security
by María Isabel Iñiguez-Luna, Jorge David Cadena-Zamudio, Marco A. Ramírez-Mosqueda, José Luis Aguirre-Noyola, Daniel Alejandro Cadena-Zamudio, Jorge Cadena-Iñiguez and Alma Armenta-Medina
BioTech 2025, 14(4), 80; https://doi.org/10.3390/biotech14040080 - 14 Oct 2025
Viewed by 1416
Abstract
Molecular docking has emerged as a pivotal computational approach in agri-food research, offering a rapid and targeted means to discover bioactive molecules for crop protection and food safety. Its ability to predict and visualize interactions between natural or synthetic compounds and specific biological [...] Read more.
Molecular docking has emerged as a pivotal computational approach in agri-food research, offering a rapid and targeted means to discover bioactive molecules for crop protection and food safety. Its ability to predict and visualize interactions between natural or synthetic compounds and specific biological targets provides valuable opportunities to address urgent agricultural challenges, including climate change and the rise in resistant crop pathogens. By enabling the in silico screening of diverse chemical entities, this technique facilitates the identification of molecules with antimicrobial and antifungal properties, specifically designed to interact with critical enzymatic pathways in plant pathogens. Recent advancements, such as the integration of molecular dynamics simulations and artificial intelligence-enhanced scoring functions, have significantly improved docking accuracy by addressing limitations like protein flexibility and solvent effects. These technological improvements have accelerated the discovery of eco-friendly biopesticides and multifunctional nutraceutical agents. Promising developments include nanoparticle-based delivery systems that enhance the stability and efficacy of bioactive molecules. Despite its potential, molecular docking still faces challenges related to incomplete protein structures, variability in scoring algorithms, and limited experimental validation in agricultural contexts. This work highlights these limitations while outlining current trends and future prospects to guide its effective application in agri-food biotechnology. Full article
(This article belongs to the Topic From Plant to Pharmacology: Understanding the Metabolism of Natural Products)
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24 pages, 2566 KB  
Review
Valorization of Second Cheese Whey Through Microalgae-Based Treatments: Advantages, Limits, and Opportunities
by Gloria Sciuto, Nunziatina Russo, Cinzia L. Randazzo and Cinzia Caggia
BioTech 2025, 14(4), 79; https://doi.org/10.3390/biotech14040079 - 9 Oct 2025
Cited by 2 | Viewed by 1468
Abstract
The dairy sector produces considerable amounts of nutrient-rich effluents, which are frequently undervalued as simple by-products or waste. In particular, Second Cheese Whey (SCW), also known as scotta, exhausted whey, or deproteinized whey, represents the liquid fraction from ricotta cheese production. Despite its [...] Read more.
The dairy sector produces considerable amounts of nutrient-rich effluents, which are frequently undervalued as simple by-products or waste. In particular, Second Cheese Whey (SCW), also known as scotta, exhausted whey, or deproteinized whey, represents the liquid fraction from ricotta cheese production. Despite its abundance and high organic and saline content, SCW is often improperly discharged into terrestrial and aquatic ecosystems, causing both environmental impact and resource waste. The available purification methods are expensive for dairy companies, and, at best, SCW is reused as feed or fertilizer. In recent years, increasing awareness of sustainability and circular economy principles has increased interest in the valorization of SCW. Biological treatment of SCW using microalgae represents an attractive strategy, as it simultaneously reduces the organic load and converts waste into algal biomass. This biomass can be further valorized as a source of proteins, pigments, and bioactive compounds with industrial relevance, supporting applications in food, nutraceuticals, biofuels, and cosmetics. This review, starting from analyzing the characteristics, production volumes, and environmental issues associated with SCW, focused on the potential of microalgae application for their valorization. In addition, the broader regulatory and sustainability aspects related to biomass utilization and treated SCW are considered, highlighting both the promises and limitations of microalgae-based strategies by integrating technological prospects with policy considerations. Full article
(This article belongs to the Special Issue A Sustainable Approach to Macroalgae and Microalgae: From Cultivation to Compound Recovery and Biotechnological Applications)
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24 pages, 13022 KB  
Article
Development of PCR Methods for Detecting Wheat and Maize Allergens in Food
by Tata Ninidze, Tamar Koberidze, Kakha Bitskinashvili, Tamara Kutateladze, Boris Vishnepolsky and Nelly Datukishvili
BioTech 2025, 14(4), 78; https://doi.org/10.3390/biotech14040078 - 1 Oct 2025
Viewed by 3762
Abstract
The detection of allergens is essential for ensuring food safety, protecting public health, and providing accurate information to consumers. Wheat (Triticum aestivum L.) and maize (Zea mays L.) are recognized as important food allergens. In this study, novel PCR methods were [...] Read more.
The detection of allergens is essential for ensuring food safety, protecting public health, and providing accurate information to consumers. Wheat (Triticum aestivum L.) and maize (Zea mays L.) are recognized as important food allergens. In this study, novel PCR methods were developed for the reliable detection of wheat and maize allergens, including wheat high-molecular-weight glutenin subunit (HMW-GS) and low-molecular-weight glutenin subunit (LMW-GS), as well as three maize allergens, namely, Zea m 14, Zea m 8, and zein. Wheat and maize genomic DNA, as well as allergen genes, were examined during 60 min of baking at 180 °C and 220 °C. Agarose gel electrophoresis revealed degradation of genomic DNA and amplified PCR fragments in correlation with increasing baking temperature and time. For each target gene, the best primers were identified that could detect HMW-GS and LMW-GS genes in wheat samples and Zea m 14, Zea m 8, and zein genes in maize samples after baking at 220 °C for 60 min and 40 min, respectively. The results indicate that these PCR methods can be used for the reliable and sensitive detection of wheat and maize allergens in processed foods. Full article
(This article belongs to the Section Industry, Agriculture and Food Biotechnology)
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15 pages, 3351 KB  
Article
Biotic Elicitor-Driven Enhancement of In Vitro Micropropagation and Organogenesis in Solanum tuberosum L. cv. Fianna
by Mario James-Forest, Ma del Carmen Ojeda-Zacarías, Alhagie K. Cham, Héctor Lozoya-Saldaña, Rigoberto E. Vázquez-Alvarado, Emilio Olivares-Sáenz and Alejandro Ibarra-López
BioTech 2025, 14(4), 77; https://doi.org/10.3390/biotech14040077 - 24 Sep 2025
Cited by 1 | Viewed by 939
Abstract
This study evaluates the impact of biotic elicitors and hormone regimes on the in vitro establishment, shoot multiplication, and organogenesis of Solanum tuberosum L. cv. Fianna under controlled laboratory conditions. Explants derived from pre-treated tubers were cultured on Murashige and Skoog (MS) medium [...] Read more.
This study evaluates the impact of biotic elicitors and hormone regimes on the in vitro establishment, shoot multiplication, and organogenesis of Solanum tuberosum L. cv. Fianna under controlled laboratory conditions. Explants derived from pre-treated tubers were cultured on Murashige and Skoog (MS) medium supplemented with vitamins and varying concentrations of growth regulators or elicitors. Aseptic establishment achieved a high success rate (~95%) using a 6% sodium hypochlorite disinfection protocol. Multiplication was significantly enhanced with a combination of 0.2 mg L−1 naphthaleneacetic acid (NAA) and 0.5–1.0 mg L−1 benzylaminopurine (BAP), producing the greatest number and length of shoots and roots. Direct organogenesis was stimulated by bio-elicitors Activane®, Micobiol®, and Stemicol® in (MS) basal medium at mid-level concentrations (0.5 g or mL L−1), improving shoot number, elongation, and root development. Activane®, Micobiol®, and Stemicol® are commercial elicitors that stimulate plant defense pathways and morphogenesis through salicylic acid, microbial, and jasmonic acid signaling mechanisms, respectively. Indirect organogenesis showed significantly higher callus proliferation in Stemicol® and Micobiol® treatments compared to the control medium, resulting in the highest fresh weight, diameter, and friability of callus. The results demonstrate the potential of biotic elicitors as alternatives or enhancers to traditional plant growth regulators in potato tissue culture, supporting more efficient and cost-effective micropropagation strategies. Full article
(This article belongs to the Section Industry, Agriculture and Food Biotechnology)
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15 pages, 1133 KB  
Article
Evaluating Scale-Up Cultivation Modes for Aspergillus oryzae Biomass Production Using VFA-Rich Effluents from Agro-Industrial Residues
by Taner Sar, Clarisse Uwineza, Mohammad J. Taherzadeh and Amir Mahboubi
BioTech 2025, 14(4), 76; https://doi.org/10.3390/biotech14040076 - 24 Sep 2025
Viewed by 1218
Abstract
Organic-waste-derived volatile fatty acids (VFAs) are promising substrates for fungal biomass cultivation, offering a nutrient-rich medium capable of meeting microbial growth requirements. However, the growth and biomass productivity are highly influenced by the VFAs’ composition and mode of operation. This study investigated the [...] Read more.
Organic-waste-derived volatile fatty acids (VFAs) are promising substrates for fungal biomass cultivation, offering a nutrient-rich medium capable of meeting microbial growth requirements. However, the growth and biomass productivity are highly influenced by the VFAs’ composition and mode of operation. This study investigated the cultivation of Aspergillus oryzae fungal biomass using agro-industrial-derived VFA effluent, employing repeated-batch and fed-batch (stepwise and continuous-feeding) cultivation modes to evaluate fungal growth and biomass composition. The highest dry biomass yield of 0.41 dry biomass/gVFAsfed (g/g) was achieved in fed-batch mode with continuous feeding, where the biomass exhibited pellet morphology, facilitating ease of harvesting. The crude protein content varied according to the cultivation strategy, reaching 45–53% in continuous-feeding fed-batch mode, while it was 34–42% in stepwise fed-batch mode. Additionally, the fungal biomass contained significant levels of essential macronutrients and trace elements, including Mg, Ca, K, Mn, and Fe, which are crucial if the biomass is intended to be used in animal feed formulations. This study highlights the effects of cultivation modes on biomass composition and the potential of VFA-derived fungal biomass as a sustainable feed ingredient. Full article
(This article belongs to the Special Issue Volatile Fatty Acids (VFAs) and Their Potential in Waste-Based Biorefineries)
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