Search Results (61)

As a pivotal model organism in Lepidoptera research, the silkworm (Bombyx mori) holds significant importance in life science due to its economic value and biotechnological applications. Advancements in proteomics and bioinformatics have enabled substantial progress in characterizing the B. mori proteome. Systematic screening and identification of protein–protein interactions (PPIs) have progressively elucidated the molecular mechanisms governing key biological processes, including viral infection, immune regulation, and growth development. This review comprehensively summarizes traditional PPI detection techniques, such as yeast two-hybrid (Y2H) and immunoprecipitation (IP), alongside emerging methodologies such as mass spectrometry-based interactomics and artificial intelligence (AI)-driven PPI prediction. We critically analyze the strengths, limitations, and technological integration strategies for each approach, highlighting current field challenges. Furthermore, we elaborate on the molecular regulatory networks of Bombyx mori nucleopolyhedrovirus (BmNPV) from multiple perspectives: apoptosis and cell cycle regulation; viral protein invasion and trafficking; non-coding RNA-mediated modulation; metabolic reprogramming; and host immune evasion. These insights reveal the dynamic interplay between viral replication and host defense mechanisms. Collectively, this synthesis aims to provide a robust theoretical foundation and technical guidance for silkworm genetic improvement, infectious disease management, and the advancement of related biotechnological applications. Full article

The black fig fly, Silba adipata McAlpine (Diptera: Lonchaeidae), is a monophagous invasive pest of fig crops. Its recent detection in Mexico has highlighted the urgent need for control strategies. However, efforts to study and manage this pest are constrained by a limited understanding of its basic biology and an inability to rear this insect in the laboratory. Some species of flies are reproductively immature at adult emergence and require specific nutrients for the development of reproductive structures. Given this, we examined the development of ovaries and testes in response to different adult diets, ovary maturation in relation to access to figs and ultraviolet (UV) light exposure and behavioral responses to fig latex. Dietary protein (hydrolyzed yeast) was essential for egg maturation. The highest prevalence of sexual maturity in females was observed at age 21 days and was not influenced by UV light or access to figs. Male testes size decreased over time irrespective of the adult diet. The consumption of latex increased when protein was not available, although the sexes differed in their response to latex over time. These findings help overcome a critical barrier to the laboratory colonization of S. adipata by demonstrating that protein-supplemented diets are essential for sexual maturation. However, the lack of information on the specific role of fig latex in the diet and the absence of sexual behavior during the experiments highlight key knowledge gaps. Future research should focus on identifying those stimuli that promote copulation and oviposition to understand the complete life cycle of this pest under controlled conditions. Full article

This work addresses the technical–economic–environmental analysis of a 1G2G ethanol integrated process using immobilized recombinant Saccharomyces cerevisiae and crude sugarcane bagasse acid hydrolysate mixed with molasses. Three case studies were evaluated and compared with the traditional 1G plants. The minimal ethanol-selling price and the life cycle assessment using CML-IA midpoint indicators were chosen as the economic and environmental metrics, respectively. The values found for the ethanol-selling price ranged from 472.92 USD/m³ to 966.53 USD/m³ for the integrated case studies. Compared to the average sales value of 1G ethanol (673.48 USD/m³), the first and second case studies were interesting for their economic viability, while the third case study would require a 43.5% increase in the price of ethanol to achieve production profitability. In the environmental assessment, the integrated 2G ethanol processes of the first and third case studies allowed for the increase in ethanol production per ton of sugarcane processed without decreasing the environmental performance of the process. The third case study presented the lowest environmental impact indicators, except for global warming potential and photochemical oxidation categories, highlighting the importance of the development of biomass pretreatment strategies with lower carbon footprint. The strategy of integrating the 2G process into a 1G ethanol biorefinery offers interesting economic and environmental values, allows the use of hemicellulose, and contributes to the development of 2G processes in sugarcane biorefineries and to the sustainability of the processes. Full article

The bioethanol produced from biomass fermentation is characterised by its renewable nature, as expressed in its life cycle. The Amazonian taro, with 72% starch content, can be an alternative to traditional raw materials such as corn and sugarcane. The objective of this study was to evaluate taro (Colocasia esculenta) as a raw material for bioethanol production through ultrasound-assisted enzymatic hydrolysis. For the enzymatic hydrolysis process, α-amylase and glucoamylase enzymes were used. Active dry yeast Saccharomyces cerevisiae was employed for fermentation. The chemical characterisation analysis of the flour with peel (WP) revealed high levels of protein (8.71%), crude fibre (4.21%) and total ash (5.56%). However, taro flour without peel (NP) revealed higher starch results (27.5%). The evaluation of reducing sugars during enzymatic hydrolysis showed better results in the samples with ultrasonic pretreatment, specifically in the sample with peel (WP-US) (1017.8 mg/L). The ethanol yield was significantly higher in the ultrasound-treated samples, achieving an approximately 35% increase in ethanol yield for both peeled and unpeeled samples compared to untreated samples. These findings open up new perspectives for using this tuber in the biofuel industry. Furthermore, they emphasise the need for continued research on biomass conversion processes. Full article

The recent coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has exerted considerable impact on global health. To prepare for rapidly mutating viruses and for the forthcoming pandemic, effective therapies targeting the critical stages of the viral life cycle need to be developed. Viruses are dependent on the interaction between the receptor-binding domain (RBD) of the viral Spike (S) protein (S-RBD) and the angiotensin-converting enzyme 2 (ACE2) receptor to efficiently establish infection and the following replicate. Targeting this interaction provides a promising strategy to inhibit the entry process of the virus, which in turn has both preventive and therapeutic effects. In this study, we developed a robust and straightforward assay based on the Yeast-Two Hybrid system (Y2H) for identifying inhibitors targeting the S-RBD-ACE2 interaction of SARS-CoV-2. Through high-throughput screening, two compounds were identified as potential entry inhibitors. Among them, IMB-1C was superior in terms of pseudovirus entry inhibition and toxicity. It could bind to both ACE2 and S-RBD and induce conformational change in the S-RBD+ACE2 complex. This is the first study to verify the feasibility of utilizing the Y2H system to discover potent SARS-CoV-2 inhibitors targeting the receptor recognition stage. This approach may also be applied in the discovery of other virus receptor recognition inhibitors. Full article

Clostridium perfringens type F is a spore-forming bacterium that causes human illnesses, including food poisoning (FP) and non-foodborne gastrointestinal diseases. In this study, we evaluated the antimicrobial activities of 15 natural products against C. perfringens spore growth. Among them, garlic, onion juice, and undiluted essential oil constituents (EOCs) of clove, rosemary, and peppermint showed the strongest activity. Therefore, we examined the inhibitory effects of these products on each stage of the life cycle of C. perfringens FP strains, including spore germination, spore outgrowth, and vegetative growth, in laboratory media and chicken meat. Both clove and peppermint oils (at 0.5%; v/v) inactivated C. perfringens spore germination in nutrient-rich trypticase–glucose–yeast extract (TGY) medium. Furthermore, EOCs at (0.1–0.5%) arrested the spore outgrowth of C. perfringens in TGY medium. Interestingly, EOCs at 0.5% completely inhibited the vegetative growth of FP isolates during a 6 h incubation in TGY medium. However, even at 4-fold higher concentrations (2%), EOCs were unable to inactivate C. perfringens spore growth in contaminated chicken meat stored under abusive conditions. Although some of the natural products inhibited C. perfringens spore germination, outgrowth, and vegetative growth in TGY medium, no such inhibitory activity was observed when these products were applied to C. perfringens spore-inoculated chicken meat. Full article

Increasing animal protein, especially from beef, is necessary to satisfy the requirements of the growing global population. However, this is becoming increasingly challenging due to stakeholders prioritizing sustainable systems with low environmental impacts. To answer these challenges, the use of feed additives such as yeast probiotics seems to be an effective route for improvements in feed efficiency and growth performance. The current study was performed to quantify the environmental impacts of the production and use of a yeast probiotic (Actisaf^® Sc 47) through an LCA method in four trials performed during the fattening period: France (French trial), Spain (Spanish trial), and Italy (Italian trials A and B). During each trial, two groups of animals with the same characteristics were compared during the fattening period: the control group without Actisaf^® Sc 47 supplementation used as a baseline, and the experimental group with Actisaf^® Sc 47 supplementation. The calves started the trial at the ages of 224, 92, and 186 days during the French, Spanish, and Italian trials A and B, respectively. Of the sixteen impact category indicators, nine were analyzed. In Italian trials A and B, the Spanish trial, and the French trial, the use of Actisaf^® Sc 47 reduced CC by 4.8, 3.8, 6.6, and 5.4%; FE by 4.8, 3.9, 8.2, and 6.3%; ME by 4.9, 4.0, 6, and 6.4%; and TE by 5.3, 4.1, and 6.2%, respectively. Moreover, during the same trials, Actisaf^® Sc 47 reduced AC by 6.0, 4.1, 6.3, and 7.1%; LU by 4.7, 3.9, 5.8, and 6.1%; WU by 4.5, 3.4, 5.2, and 5.9%; RU (fossils) by 4.8, 3.7, 7.3, and 4.8%; and PM by 5.2, 4.1, 6.3, and 7.1%, respectively. The use of Actisaf^® Sc 47 has beneficial effects on the environment. Regarding Actisaf^® Sc 47 production, the results showed that the production of 1 kg of Actisaf^® Sc 47 emitted 2.1 kg of CO₂ eq, 60% of which came from the fermentation process and 39% from drying. The results demonstrate the interest in using Actisaf^® Sc during the fattening period for its multiple positive outcomes: improving performance and efficiency, while reducing the global carbon footprint. Full article

In this study, butter and the corresponding buttermilk samples were produced with cream fermented by aromatic (A) or probiotic (P) cultures with or without complementary bioprotective culture (BC). The samples were characterised for their composition and colour parameters. Texture and rheological properties were evaluated at 10 and 20 °C. Microbiological (lactobacilli, lactococci, and yeast and mould counts) and sensory (aroma, taste, texture, and global evaluation) analyses were also performed. All butter sample characteristics were in accordance with the Portuguese standard. Regarding colour, the sample obtained with cream fermented by probiotics plus bioprotective culture (PBC) presented higher L* and b* values, indicating a slightly higher yellow chroma. However, colour differences (ΔEab*) in the butter samples were, in most cases, not detectable by a common observer. Butter samples P and PBC presented a significantly higher viscous modulus and consequently higher dynamic viscosity values (ca. log 6.5 Pa.s at 10 °C and log 5 Pa.s at 20 °C). Butter samples presented a pseudoplastic behaviour, and rheological parameters showed a high dependence on temperature. The counts of lactobacilli and lactococci in the butter samples were of the order of log 7–8 CFU/g, while yeast and mould counts were lower than log 2 CFU/g until the 30th day of storage, after which they showed a sharp increase to ca. log 5 CFU/g between the 30th and the 60th days of storage. Regarding sensory attributes of butter, sample P received the highest overall liking, followed by samples ABC and PBC. Sample A was the least appreciated. Buttermilk samples presented significant differences regarding their composition, viscosity, and colour parameters. In all cases, lactobacilli and lactococci counts exceeded log 7 CFU/mL after 30 days of storage, but yeast and mould counts were of the order of log 5–6 CFU/mL at the 15th day of storage. Samples P and PBC presented yeast and mould counts ca. 1–2 log cycles lower than samples A and ABC, indicating the potential of probiotic and bioprotective cultures to extend the shelf life of the product. Regarding the sensory attributes of buttermilk, samples P and PBC received the highest overall liking, followed by sample ABC. Sample A received the lowest scores, as had occurred with the butter samples. However, in all cases, the scores obtained by the buttermilk samples were lower compared to the ones of the corresponding butter. It can be concluded that both probiotic butter and buttermilk present high levels of lactobacilli and lactococci and can maintain their probiotic potential throughout the storage period. Full article

Repurposing saffron (Crocus sativus) waste presents a sustainable strategy for generating high-value products within the bioeconomy framework. Typically, flower components are discarded after stigma harvest, resulting in significant waste—350 kg of tepals per kilogram of stigmas. This research employed a comprehensive approach, integrating bioactivity studies (in vitro and in silico) with Life Cycle Assessment (LCA) evaluations, to extract and assess bioactive compounds from C. sativus tepals sourced in Tuscany, Italy. Phytochemical characterization using UPLC-MS/MS revealed a high abundance and variety of flavonoids in the hydro-ethanolic extract (CST). The antioxidant capacity was validated through various assays, and the ability to mitigate H₂O₂-induced oxidative stress and enhance fermentation was demonstrated in Saccharomyces cerevisiae. This study reports that C. sativus tepals extract reduces oxidative stress and boosts ethanol fermentation in yeast, paving the way for applications in the food and biofuels sectors. Further validation in RAW 264.7 macrophages confirmed CST’s significant anti-inflammatory effects, indicating its potential for pharmaceutical, cosmeceutical, and nutraceutical applications. In silico studies identified potential targets involved in antioxidant and anti-inflammatory processes, shedding light on possible interaction mechanisms with Kaempferol 3-O-sophoroside (KOS-3), the predominant compound in the extract. The integration of LCA studies highlighted the environmental benefits of this approach. Overall, this research underscores the value of using waste-derived extracts through “green” methodologies, offering a model that may provide significant advantages for further evaluations compared to traditional methodologies and supporting the circular bioeconomy. Full article

Rad6 is a canonical ubiquitin-conjugating enzyme known for its role in regulating chromosome-related cellular processes in yeast and has been proven to have multiple functions in Beauveria bassiana, including insect-pathogenic lifestyle, UV damage repair, and conidiation. However, previous studies have only reported the key role of Rad6 in regulating conidial production in a nutrient-rich medium, without any deep mechanism analyses. In this study, we found that the disruption of Rad6 leads to a profound reduction in conidial production, irrespective of whether the fungus is cultivated in nutrient-rich or nutrient-poor environments. The absence of rad6 exerts a suppressive effect on the transcription of essential genes in the central developmental pathway, namely, brlA, abaA, and wetA, resulting in a direct downregulation of conidiation capacity. Additionally, mutant strains exhibited a more pronounced decline in both conidial generation and hyphal development when cultured in nutrient-rich conditions. This observation correlates with the downregulation of the central developmental pathway (CDP) downstream gene vosA and the upregulation of flaA in nutrient-rich cultures. Moreover, single-transcriptomics analyses indicated that irregularities in biotin metabolism, DNA repair, and tryptophan metabolism are the underlying factors contributing to the reduced conidial production. Comprehensive dual transcriptomics analyses pinpointed abnormal biotin metabolism as the primary cause of conidial production decline. Subsequently, we successfully restored conidial production in the Rad6 mutant strain through the supplementation of biotin, further confirming the transcriptomic evidence. Altogether, our findings underscore the pivotal role of Rad6 in influencing biotin metabolism, subsequently impacting the expression of CDP genes and ultimately shaping the asexual life cycle of B. bassiana. Full article

Today, one of the major challenges of dairy farmers is to reduce their environmental footprint to establish more effective, efficient, and sustainable production systems. Feed additives such as yeast probiotics could potentially allow them to achieve these objectives through the improvement of milk production, feed efficiency, and ration valorization, hence mitigating the environmental impacts of milk production. In this study, the life cycle assessment (LCA) principle was performed to estimate the environmental impact of the production and supplementation of a commercial yeast probiotic (Actisaf Sc 47) in three trials performed in three different countries that are representative for around 50% of the milk production in Europe: France (French trial), United Kingdom (UK trial), and Germany (German trial). For each trial, two groups of animals were compared: control, without Actisaf Sc 47 supplementation, used as baseline; and experimental, with Actisaf Sc 47 supplementation at 5 or 10 g/cow/day. Different impact categories were analyzed for each group to calculate the impact of producing 1 kg of fat- and protein-corrected milk. An initial analysis was done only during the period of Actisaf Sc 47 supplementation and showed than the supplementation with Actisaf Sc 47 reduced, on average by 5%, the carbon footprint during the three trials. A second analysis was done via the extrapolation of all the data of each trial to an annual farm level, including the lactation period (305 days), dry period (60 days), and the period with and without Actisaf Sc 47 supplementation. Reported at a farm annual scale, the average reduction allowed by Actisaf Sc 47 supplementation was 2.9, 2.05, 2.47, 1.67, 2.28, 2.18, 2.14, and 2.28% of the carbon footprint, land use, water use, resource use, acidification, freshwater eutrophication, marine eutrophication, and terrestrial eutrophication, respectively. On average, the production of 1 kg of fat- and protein-corrected milk by using Actisaf Sc 47 was shown to improve environmental impacts compared to control. Regarding Actisaf Sc 47 production, the LCA showed that the production of 1 kg of Actisaf Sc 47 emitted 2.1 kg CO₂ eq with a negligible contribution to total the carbon footprint of milk ranging from 0.005 to 0.016%. The use of Actisaf Sc 47 in dairy cows could then result in different positive outcomes: improving performance and efficiency while reducing the global carbon footprint. Full article

Dietary supplementation of yeast-derived mannan-rich fraction (MRF) could improve the gastrointestinal health and production efficiency of broilers, and, consequently, lower the environmental impacts of chicken production. The objective of this meta-analysis was to quantify the retrospective effects of feeding MRF (Actigen^®, Alltech Inc., Nicholasville, KY) on the production performance of broilers. The meta-analysis database included 27 studies and consisted of 66 comparisons of MRF-supplemented diets vs. basal (i.e., negative control) and antibiotic-supplemented (i.e., positive control) diets. A total of 34,596 broilers were involved in the comparisons and the average final age of the birds was 35 days. Additionally, the impact of feeding MRF on the carbon footprint (feed and total emission intensities) of chicken production was evaluated using the meta-analysis results of broiler performance (MRF vs. basal diets) to develop a scenario simulation that was analyzed by a life cycle assessment (LCA) model. A database of all trials (MRF vs. basal and antibiotic diets) indicated that feeding MRF increased (p < 0.01) average daily feed intake (ADFI; +3.7%), final body weight (FBW; +3.5%), and average daily gain (ADG; 4.1%) and improved (p < 0.01) feed conversion ratio (FCR; −1.7%) without affecting (p > 0.05) mortality. A subdatabase of MRF vs. basal diets indicated that dietary MRF increased ADFI (+4.5%), FBW (+4.7%), and ADG (+6.3%) and improved FCR (−2.2%) and mortality (−21.1%). For the subdatabase of MRF vs. antibiotic diets, both treatments exhibited equivalent effects (p > 0.05) on broiler performance parameters, suggesting that MRF could be an effective alternative to in-feed antibiotics. Subgroup analysis revealed that different study factors (year of study, breed/strain, production challenges, and MRF feeding duration) influenced the effect of dietary MRF on broiler performance. Simulated life cycle analysis (LCA) indicated that feeding MRF decreased feed and total emission intensities, on average, by −2.4% and −2.1%, respectively. In conclusion, these results demonstrate that dietary MRF is an effective nutritional solution for improving broiler performance, an effective alternative to in-feed antibiotic growth promoters, and reduces the environmental impact of poultry meat production. Full article

Bovine whey cheese (WC) is a product from southern European countries that presents some challenges: its production process involves high energy inputs; the yield is low; and WC has a short shelf life. The application of ultrafiltration (UF) to bovine whey before manufacture of WC and the employment of protective cultures can reduce these disadvantages. The objective of this research was the production of whey cheeses using ultrafiltrated bovine cheese whey with added probiotics or probiotics plus protective cultures. Three types of WC were produced: control CW without any addition (C); CW with the addition of the probiotic Lactobacillus acidophilus (LA5); and CW with the addition of Lactobacillus acidophilus plus a protective culture containing Lacticaseibacillus paracasei and Lacticaseibacillus rhamnosus (LA5FQ4). The WCs were stored under refrigerated conditions for 28 days. The products with added cultures presented lower pH values and higher titratable acidities when compared to the control. Sample LA5 presented the lowest pH and the highest titratable acidity, while LA5FQ4 presented intermediate values. Slight differences were observed between products regarding color parameters, chiefly resulting from storage time. The samples with added cultures were firmer when compared to the control, with LA5 cheeses showing the highest values at the end of the storage. Lactic acid bacteria (LAB) counts were on the order of log 8–9 CFU/g for the products with added cultures. Lower levels of yeasts and molds were detected on the sample with the protective culture (LA5FQ4), so that by the end of storage they presented counts one log cycle lower than C and LA5. Hence, the beneficial impact of the protective culture on the shelf life of the product is evident. Regarding sensory evaluation, LA5FQ4 cheeses obtained the highest scores for all parameters evaluated. It can be concluded that the use of UF associated with the addition of protective cultures can be very useful to reduce the energy consumption of the manufacturing process, to prolong the shelf life of WC and to improve its sensory properties. Full article

Chaperonin containing tailless complex polypeptide 1 (CCT) is a molecular chaperone protein that consists of eight completely different subunits and assists in the folding of newly synthesized peptides. The zeta subunit of CCT is a regulatory factor for the folding and assembly of cytoskeletal proteins as individuals or complexes. In this study, the zeta subunit of Nosema bombycis (NbCCTζ) is identified for the first time. The complete ORF of the NbCCTζ gene is 1533 bp in length and encodes a 510 amino acid polypeptide. IFA results indicate that NbCCTζ is colocalized with actin and β-tubulin in the cytoplasm during the proliferative phase and that NbCCTζ is completely colocalized with NbCCTα in the cytoplasm of N. bombycis throughout the entire life cycle. Furthermore, the yeast two-hybrid assay revealed that the NbCCTζ interacts with NbCCTα. The transcriptional level of NbCCTζ is significantly downregulated by knocking down the NbCCTα gene, while the transcriptional level of NbCCTα is downregulated after knocking down the NbCCTζ gene. These results suggest that NbCCTζ may play a vital role in the proliferation of N. bombycis by coordinating with NbCCTα. Full article

Reversed-flow gas chromatography (R.F.G.C.) was employed to assess the impact of genetic modification on Saccharomyces cerevisiae yeast strains during the process of alcoholic fermentation, utilizing fig syrup. Multiple fermentations were carried out at various temperatures to evaluate the influence of genetic modifications on yeast strain efficiency. The study involved a wild-type yeast strain, W303, as a control and two genetically modified strains, W_M4_533 and W_M4_558, sharing the same genetic background as the wild type. Notably, the genetic modifications in the Msn4p transcription factor involved the substitution of serine residues with alanine at positions 533 and 558, resulting in the development of psychrophilic or ethanol-resistant strains. Utilizing the R.F.G.C. method enabled the differentiation of the duration of alcoholic fermentation phases, providing insights correlated to the yeast cell life cycle. The values of rate constants (k) for each phase, conducted with both wild-type and genetically modified cells using RFGC, aligned with the existing literature. Additionally, the calculation of activation energies for distinct phases revealed lower values for genetically modified strains compared to wild-type strains. This decrease in activation energies suggests enhanced efficiency in the alcoholic fermentation process for the genetically modified strains. Full article