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Lutein is widely used in medicine, health care, and food processing due to its antioxidant effects; however, it is difficult for the traditional extraction of lutein using marigolds to meet the increasing market demand for lutein. To achieve high-efficiency lutein production, we investigated the effects of different conditions on the biomass accumulation and lutein yield of Chlorella vulgaris. The optimized cultivation conditions include mixotrophic cultivation using sodium nitrate as a nitrogen source, maintaining a total-organic-carbon-to-total-nitrogen ratio of 12:1, a total-nitrogen-to-total-phosphorus ratio of 10:1, and lighting duration of 24 h. The results of the study indicated that under these specific conditions, Chlorella vulgaris attained a final biomass concentration, biomass productivity, and growth yield of 6.08 g·L−1, 1.00 g·L−1·d−1, and 1.67 g biomass/g TOC, respectively. Additionally, the concentrations of total chlorophyll, carotenoid, lutein, and protein reached 139.20 mg·L−1, 31.87 mg·L−1, 15.02 mg·L−1, and 2.17 g·L−1, respectively, and the content of lutein reached 2.47 mg·g−1. This study supplies a theoretical basis for the industrial application of lutein production using Chlorella vulgaris.

Aggressive forms of periodontitis, especially in young patients, are often associated with an increased proportion of the Gram-negative bacterium Aggregatibacter actinomycetemcomitans of the microbiota of the affected periodontal sites. One of the virulence factors of A. actinomycetemcomitans is a leukotoxin (LtxA) that induces a pro-inflammatory cell death process in leukocytes. A. actinomycetemcomitans exhibits a large genetic diversity and different genotypes vary in LtxA production capacity. The genotype JP2 is a heavy LtxA producer due to a 530-base pair deletion in the promoter for the toxin genes, and this trait has been associated with an increased pathogenic potential. The present study focused on the production and release of LtxA by different A. actinomycetemcomitans genotypes and serotypes under various growth conditions. Four different strains of this bacterium were cultured in two different culture broths, and the amount of LtxA bound to the bacterial surface or released into the broths was determined. The cultures were examined during the logarithmic and the early stationary phases of growth. The JP2 genotype exhibited the highest LtxA production among the strains tested, and production was not affected by the growth phase. The opposite was observed with the other strains. The composition of the culture broth had no effect on the growth pattern of the tested strains. However, the abundant release of LtxA from the bacterial surface into the culture broth was found in the presence of horse serum. Besides confirming the enhanced leucotoxicity of the JP2 genotype, the study provides new data on LtxA production in the logarithmic and stationary phases of growth and the effect of media composition on the release of the toxin from the bacterial membrane.

Tilapia lake virus (TiLV) presents a substantial threat to global tilapia production. Despite the development of numerous cell lines for TiLV isolation and propagation, none have been specifically derived from red hybrid tilapia (Oreochromis spp.). In this study, we successfully established a new cell line, RHTiB, from the red hybrid tilapia brain. RHTiB cells were cultured for 1.5 years through over 50 passages and demonstrated optimal growth at 25 °C in Leibovitz-15 medium supplemented with 10% fetal bovine serum at pH 7.4. Morphologically, RHTiB cells displayed a fibroblast-like appearance, and cytochrome oxidase I gene sequencing confirmed their origin from Oreochromis spp. Mycoplasma contamination testing yielded negative results. The revival rate of the cells post-cryopreservation was observed to be between 75 and 80% after 30 days. Chromosomal analysis at the 25th passage revealed a diploid count of 22 pairs (2n = 44). While no visible cytopathic effects were observed, both immunofluorescence microscopy and RT-qPCR analysis demonstrated successful TiLV propagation in the RHTiB cell line, with a maximum TiLV concentration of 107.82 ± 0.22 viral copies/400 ng cDNA after 9 days of incubation. The establishment of this species-specific cell line represents a valuable advancement in the diagnostic and isolation tools for viral diseases potentially impacting red hybrid tilapia.

Hydrogen is ideal for replacing fossil fuels because upon combustion it generates only water. Dark fermentation (DF) from lignocellulose might be a competitive process for hydrogen production at the industrial scale. However, lignocellulose must be pretreated to obtain fermentable sugars, which is costly and creates pollution. Microorganisms from bovine rumen efficiently degrade lignocellulose. Unfortunately, they have scarcely been explored for the production of hydrogen. Therefore, deeper studies on the culture conditions have to be undertaken to understand the behavior of microbial consortia from the rumen of bovines (MCRB) during hydrogen production. In this work, we evaluated the production of hydrogen by DF with MCRB by varying the incubation time, two culture media (MB and Rhodospirillaceae), headspace (40 and 80 mL), and thermal treatment. It was found that the production of hydrogen was maximum at 16 h MCRB incubation in MB. An amount of 80 mL headspace resulted in a threefold production of hydrogen as compared to 40 mL; the MCRB without heat treatment had a higher H2 yield. The production of hydrogen with 32 MCRB was highly variable, ranging between 21 and 696 mL. Our findings show a different perspective on the treatment of MCRB for the production of hydrogen and give insights on the impact of the culture conditions for increasing hydrogen production.

Worldwide, bacterial resistance is one of the most severe public health problems. Currently, the failure of antibiotics to counteract superbugs highlights the need to search for new molecules with antimicrobial potential to combat them. The objective of this research was to evaluate the antimicrobial activity of Bacillus amyloliquefaciens BS4 against Gram-negative bacteria. Thirty yeasts and thirty-two Bacillus isolates were tested following the agar well-diffusion method. Four Bacillus sp. strains (BS3, BS4, BS17, and BS21) showed antagonistic activity against E. coli ATCC 25922 using bacterial culture (BC) and the cell-free supernatant (CFS), where the BS4 strain stood out, showing inhibitory values of 20.50 ± 0.70 mm and 19.67 ± 0.58 mm for BC and CFS, respectively. The Bacillus sp. BS4 strain can produce antioxidant, non-hemolytic, and antimicrobial metabolites that exhibit activity against several microorganisms such as Salmonella enterica, Klebsiella pneumoniae, Shigella flexneri, Enterobacter aerogenes, Proteus vulgaris, Yersinia enterocolitica, Serratia marcescens, Aeromonas sp., Pseudomonas aeruginosa, Candida albicans, and Candida tropicalis. According to the characterization of the supernatant, the metabolites could be proteinaceous. The production of these metabolites is influenced by carbon and nitrogen sources. The most suitable medium to produce antimicrobial metabolites was TSB broth. The one-factor-at-a-time method was used to standardize parameters such as pH, agitation, temperature, carbon source, nitrogen source, and salts, resulting in the best conditions of pH 7, 150 rpm, 28 °C, starch (2.5 g/L), tryptone (20 g/L), and magnesium sulfate (0.2 g/L), respectively. Moreover, the co-culture was an excellent strategy to improve antimicrobial activity, achieving maximum antimicrobial activity with an inhibition zone of 21.85 ± 1.03 mm. These findings position the Bacillus amyloliquefaciens BS4 strain as a promising candidate for producing bioactive molecules with potential applications in human health.

Manglietiastrum sinicum Y.W. Law is a critically endangered species with great ornamental and commercial value, which urgently requires protection. We tested different combinations of basal media and plant growth regulators to determine (i) the optimal conditions for bud induction and proliferation of explants and (ii) optimal rooting conditions. RAPD- and ISSR-PCR were used to assess the genetic fidelity of regenerated plantlets. Murashige and Skoog medium (MS) supplemented with 0.5 mg/L 6-benzyladenine (BA) and 0.05 mg/L indole-3-butyric acid (IBA) is the optimal medium for bud induction (100% induction). MSM medium (a special basal medium for M. sinicum) was more suitable for the efficient proliferation and rooting of M. sinicum. Maximum bud proliferation rate (446.20%) was obtained on MSM, with 0.4 mg/L BA, 0.5 mg/L kinetin, and 0.06 mg/L IBA, while maximum root induction rate (88.89%) was obtained on MSM supplemented with 0.4 mg/L 1-naphthylacetic acid and 1.0 mg/L IBA with a 7-day initial darkness treatment. The rooted plantlets were transferred to a substrate containing peat soil, perlite, coconut chaff, and bark (volume ratio 2:1:1:1), with a resulting survival rate of 92.2%. RAPD and ISSR markers confirmed the genetic uniformity and stability of regenerated plants.

This review examines the production of the microbial polysaccharide gellan, synthesized by Sphingomonas elodea, on dairy and plant-based processing coproducts. Gellan is a water-soluble gum that structurally exists as a tetrasaccharide comprised of 20% glucuronic acid, 60% glucose and 20% rhamnose, for which various food, non-food and biomedical applications have been reported. A number of carbon and nitrogen sources have been tested to determine whether they can support bacterial gellan production, with several studies attempting to optimize gellan production by varying the culture conditions. The genetics of the biosynthesis of gellan has been explored in a number of investigations and specific genes have been identified that encode the enzymes responsible for the synthesis of this polysaccharide. Genetic mutants exhibiting overproduction of gellan have also been identified and characterized. Several dairy and plant-based processing coproducts have been screened to learn whether they can support the production of gellan in an attempt to lower the cost of synthesizing the microbial polysaccharide. Of the processing coproducts explored, soluble starch as a carbon source supported the highest gellan production by S. elodea grown at 30 °C. The corn processing coproducts corn steep liquor or condensed distillers solubles appear to be effective nitrogen sources for gellan production. It was concluded that further research on producing gellan using a combination of processing coproducts could be an effective solution in lowering its overall production costs.

Exopolysaccharides are important metabolites of Monascus with healthy activities. However, the low production level limits their applications. Hence, the aim of this work was to increase the yield of exopolysaccharides (EPS) and optimize liquid fermentation by adding flavonoids. The EPS yield was optimized via both medium composition and culture conditions. The optional fermentation conditions achieved for EPS production of 7.018 g/L were 50 g/L sucrose, 3.5 g/L yeast extract, 1.0 g/L MgSO4·7H2O, 0.9 g/L KH2PO4, 1.8 g/L K2HPO4·3H2O, 1 g/L quercetin, and 2 mL/L Tween-80, with pH 5.5, inoculum size 9%, seed age 52 h, shaking speed 180 rpm, and fermentation culture 100 h, respectively. Furthermore, the addition of quercetin increased EPS production by 11.66%. The results also showed little citrinin residue in the EPS. The exopolysaccharides’ composition and antioxidant capacity of quercetin-modified exopolysaccharides were then preliminarily investigated. The addition of quercetin changed the composition of the exopolysaccharides and the molecular weight (Mw). In addition, the antioxidant activity of Monascus exopolysaccharides was monitored using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS+), and -OH. Monascus exopolysaccharides have good scavenging ability of DPPH and -OH. Furthermore, quercetin increased the scavenging ABTS+ ability. Overall, these findings provide a potential rationale for the application of quercetin in improving the EPS yield.

Pluripotent stem cells (PSCs) are characterized by the ability to self-renew as well as undergo multidirectional differentiation. Culture conditions have a pivotal influence on differentiation pattern. In the current study, we compared the fate of mouse PSCs using two culture media: (1) chemically defined, free of animal reagents, and (2) standard one relying on the serum supplementation. Moreover, we assessed the influence of selected regulators (WNTs, SHH) on PSC differentiation. We showed that the differentiation pattern of PSCs cultured in both systems differed significantly: cells cultured in chemically defined medium preferentially underwent ectodermal conversion while their endo- and mesodermal differentiation was limited, contrary to cells cultured in serum-supplemented medium. More efficient ectodermal differentiation of PSCs cultured in chemically defined medium correlated with higher activity of SHH pathway while endodermal and mesodermal conversion of cells cultured in serum-supplemented medium with higher activity of WNT/JNK pathway. However, inhibition of either canonical or noncanonical WNT pathway resulted in the limitation of endo- and mesodermal conversion of PSCs. In addition, blocking WNT secretion led to the inhibition of PSC mesodermal differentiation, confirming the pivotal role of WNT signaling in this process. In contrast, SHH turned out to be an inducer of PSC ectodermal, not mesodermal differentiation.

Cyanobacteria are microorganisms able to adapt to a wide variety of environmental conditions and abiotic stresses. They produce a large number of metabolites that can participate in the dynamic adaptation of cyanobacteria to a range of different light, temperature, and nutrient conditions. Studying the metabolite profile is one way to understand how the physiological status of cells is related to their adaptive response. In this study, we sought to understand how the diversity and dynamics of the whole metabolome depended on the growth phase and various abiotic factors such as light intensity and temperature. The cyanobacterium, Aliinostoc sp. PMC 882.14, was selected for its large number of biosynthetic gene clusters. One group of cells was grown under normal conditions as a control, while other groups were grown under higher light or temperature. Metabolomes were analyzed by mass spectrometry (qTOF-MS/MS) combined with untargeted analysis to investigate metabolite dynamics, and significant variation was found between exponential and stationary phases, regardless of culture conditions. In the higher light group, the synthesis of several metabolites, including shinorine, was induced while other metabolites, such as microviridins, were synthesized under higher temperature conditions. Among highly regulated metabolites, we observed the presence of mycosporine-like amino acids (MAAs) and variants of somamides, microginins, and microviridins. This study demonstrated the importance of considering the physiological state of cyanobacteria for comparative global metabolomics and studies of the regulatory processes involved in production of specific metabolites. Our results also open up new perspectives on the use of organisms such as cyanobacteria for the targeted production of bioactive metabolites.

This study describes a successful protocol for establishing cell lines from the threatened Triturus cristatus in terms of collection, preparing, establishing, cryopreserving, thawing and quality checking. Different parameters such as media, media change, fresh vs. cryopreserved tissue and seeding density were tested to optimize culture conditions for this species. With fresh tissue, no considerable differences in the use of two different media were found, but with cryopreserved tissue, a combination of ITS (insulin/transferrin/selenite) and 2-mercaptoethanol had a positive effect on growth. Real-time measurements on the cell lines were used, for the first time in amphibian cells, to investigate the effect of different treatments such as media change with or without washing. Media change had a positive impact on the cells, whereas the effect was negative when combined with washing. It is concluded that establishment of cell lines is possible from the great crested newt, especially when using fresh tissue, but much more challenging if the tissue has been cryopreserved. Real-time measurement during cell culture is a useful tool to visualize the sensitivity of amphibian cells during different culture treatments.

Antibacterial peptides or bacteriocins produced by many strains of lactic acid bacteria have been used as food preservatives for many years without any known adverse effects. Bacteriocin titres can be modified by altering the physiological and nutritional factors of the producing bacterium to improve the production in terms of yield and productivity. The effects of culture conditions (initial pH, inoculum age and inoculum size) and medium compositions (organic and inorganic nitrogen sources; carbon sources) were assessed for the production of bacteriocin-like inhibitory substances (BLIS) by Lactococcus lactis Gh1 in shake flask cultures. An inoculum of the mid-exponential phase culture at 1% (v/v) was the optimal age and size, while initial pH of culture media at alkaline and acidic state did not show a significant impact on BLIS secretion. Organic nitrogen sources were more favourable for BLIS production compared to inorganic sources. Production of BLIS by L. lactis Gh1 in soytone was 1.28-times higher as compared to that of organic nitrogen sources ((NH4)2SO4). The highest cell concentration (XmX = 0.69 ± 0.026 g·L−1) and specific growth rate (μmax = 0.14 h−1) were also observed in cultivation using soytone. By replacing carbon sources with fructose, BLIS production was increased up to 34.94% compared to BHI medium, which gave the biomass cell concentration and specific growth rate of 0.66 ± 0.002 g·L−1 and 0.11 h−1, respectively. It can be concluded that the fermentation factors have pronounced influences on the growth of L. lactis Gh1 and BLIS production. Results from this study could be used for subsequent application in process design and optimisation for improving BLIS production by L. lactis Gh1 at larger scale.

Umbilical cord mesenchymal stromal cells (UCMSCs) have shown an ability to modulate the immune system through the secretion of paracrine mediators, such as extracellular vesicles (EVs). However, the culture conditions that UCMSCs are grown in can alter their secretome and thereby affect their immunomodulatory potential. UCMSCs are commonly cultured at 21% O2 in vitro, but recent research is exploring their growth at lower oxygen conditions to emulate circulating oxygen levels in vivo. Additionally, a pro-inflammatory culture environment is known to enhance UCMSC anti-inflammatory potential. Therefore, this paper examined EVs from UCMSCs grown in normal oxygen (21% O2), low oxygen (5% O2) and pro-inflammatory conditions to see the impact of culture conditions on the EV profile. EVs were isolated from UCMSC conditioned media and characterised based on size, morphology and surface marker expression. EV protein cargo was analysed using a proximity-based extension assay. Results showed that EVs had a similar size and morphology. Differences were found in EV protein cargo, with pro-inflammatory primed EVs showing an increase in proteins associated with chemotaxis and angiogenesis. This showed that the UCMSC culture environment could alter the EV protein profile and might have downstream implications for their functions in immunomodulation.

Overproduction of biomass and hyper-accumulation of lipids endow microalgae with promising characteristics to realize the cost-effective potential of advanced bioenergy. This study sought to heuristically optimize the culture conditions on a rarely reported Golenkinia sp. The results indicate that Golenkinia SDEC-16 can withstand the strong light intensity and grow in a modified BG11 medium. The optimal culture conditions for the favorable tradeoff between biomass and lipid accumulation were suggested as follows, 25,000 lux of light intensity, 9 mM of initial nitrogen concentration, and 20 mM of initial sodium chloride concentration. Under these conditions, the biomass concentration and productivity reached 6.65 g/L and 545 mg/L/d, and the synchronous lipid content and productivity reached 54.38% and 296.39 mg/L/d. Hypersalinity significantly promoted lipid contents at the cost of biomass and resulted in an increase of cell size but loss of spines of Golenkinia SDEC-16. The results shed new light on optimizing biomass and lipid productivity.

This review examines the production of the biopolymer curdlan, synthesized by Agrobacterium species (sp.), on processing coproducts and plant lignocellulosic hydrolysates. Curdlan is a β-(13)-D-glucan that has various food, non-food and biomedical applications. A number of carbon sources support bacterial curdlan production upon depletion of nitrogen in the culture medium. The influence of culture medium pH is critical to the synthesis of curdlan. The biosynthesis of the β-(13)-D-glucan is likely controlled by a regulatory protein that controls the genes involved in the bacterial production of curdlan. Curdlan overproducer mutant strains have been isolated from Agrobacterium sp. ATCC 31749 and ATCC 31750 by chemical mutagenesis and different selection procedures. Several processing coproducts of crops have been utilized to support the production of curdlan. Of the processing coproducts investigated, cassava starch waste hydrolysate as a carbon source or wheat bran as a nitrogen source supported the highest curdlan production by ATCC 31749 grown at 30 °C. To a lesser extent, plant biomass hydrolysates have been explored as possible substrates for curdlan production by ATCC 31749. Prairie cordgrass hydrolysates have been shown to support curdlan production by ATCC 31749 although a curdlan overproducer mutant strain, derived from ATCC 31749, was shown to support nearly double the level of ATCC 31749 curdlan production under the same growth conditions.

Genus Mentha, a member of Lamiaceae family, encompasses a series of species used on an industrial scale and with a well-described and developed culture process. Extracts of this genus are traditionally used as foods and are highly valued due to the presence of significant amounts of antioxidant phenolic compounds. Many essential oil chemotypes show distinct aromatic flavor conferred by different terpene proportions. Mint extracts and their derived essential oils exert notable effects against a broad spectrum of bacteria, fungi or yeasts, tested both in vitro or in various food matrices. Their chemical compositions are well-known, which suggest and even prompt their safe use. In this review, genus Mentha plant cultivation, phytochemical analysis and even antimicrobial activity are carefully described. Also, in consideration of its natural origin, antioxidant and antimicrobial properties, a special emphasis was given to mint-derived products as an interesting alternative to artificial preservatives towards establishing a wide range of applications for shelf-life extension of food ingredients and even foodstuffs. Mentha cultivation techniques markedly influence its phytochemical composition. Both extracts and essential oils display a broad spectrum of activity, closely related to its phytochemical composition. Therefore, industrial implementation of genus Mentha depends on its efficacy, safety and neutral taste.

Violaxanthin is a major xanthophyll pigment in the microalga Eustigmatos cf. polyphem, but the amount produced after propagation can vary depending upon culture conditions. In this study, the effects of cultivation time, nitrogen concentration, light intensity, and culture mode on violaxanthin production were investigated. The results showed that this microalga vigorously grew and maintained a high level of violaxanthin in the fed-batch culture, and the highest violaxanthin productivity of 1.10 ± 0.03 mg L−1 d−1 was obtained under low light illumination with 18 mM of initial nitrogen supply for ten days. Additionally, violaxanthin was purified from E. cf. polyphem by silica gel chromatography and preparative high-performance liquid chromatography (PHPLC), and identified with high-resolution mass spectrometry (HRMS). The antioxidant activity of the purified violaxanthin was evaluated by three tests in vitro: reducing power assay, 2,2-diphenyl-1-picrylhydrazyl (DPPH), and 2,2-azobis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) radical assays. The strongest inhibition of purified violaxanthin occurred during the scavenging of ABTS+ radicals, with EC50 of 15.25 μg mL−1. In conclusion, this is the first report to investigate the effects of different culture conditions on violaxanthin accumulation in E. cf. polyphem and provide a novel source for the production of violaxanthin that can be used for food and pharmaceutical applications.

Natural polysaccharides have received much attention due to their wide range of applications. Although most microbial exopolysaccharides (EPSs) use sugars as the major carbon source, such as glucose or sucrose, in this study, EPSs were induced from a squid pen powder (SPP)-containing medium by Paenibacillus mucilaginosus TKU032, a bacterial strain isolated from Taiwanese soil. Under the optimal culture conditions, the maximum EPS yield (14.8 g/L) was obtained. MALDI-TOF MS analysis of an EPS fraction purified by gel filtration revealed two mass peaks with molecular weights of ∼1.05 × 104 and ∼1.35 × 104 Da, respectively. The analysis of the hydrolysates of TKU032 EPS with cellulase, pectinase or α-amylase indicated that the glycosidic bond of TKU032 EPS is most likely an α-1,4 glycosidic bond and the hydrolysates are similar to those of starch. In addition, the purified EPS demonstrated strong antioxidant abilities.