Alternative Culture Medium Research Articles

Perspective of Spirulina culture with wastewater into a sustainable circular bioeconomy

Spirulina biomass accounts for 30% of the total algae biomass production globally. In conventional process of Spirulina biomass production, cultivation using chemical-based culture medium contributes 35% of the total production cost. Moreover, the environmental impact of cultivation stage is the highest among all the production stages which resulted from the extensive usage of chemicals and nutrients. Thus, various types of culture medium such as chemical-based, modified, and alternative culture medium with highlights on wastewater medium is reviewed on the recent advances of culture media for Spirulina cultivation. Further study is needed in modifying or exploring alternative culture media utilising waste, wastewater, or by-products from industrial processes to ensure the sustainability of environment and nutrients source for cultivation in the long term. Moreover, the current development of utilising wastewater medium only support the growth of Spirulina however it cannot eliminate the negative impacts of wastewater. In fact, the recent developments in coupling with wastewater treatment technology can eradicate the negative impacts of wastewater while supporting the growth of Spirulina. The application of Spirulina cultivation in wastewater able to resolve the global environmental pollution issues, produce value added product and even generate green electricity. This would benefit the society, business, and environment in achieving a sustainable circular bioeconomy.

Alternative culture media and cold-drying for obtaining high biological value Arthrospira platensis (Cyanobacteria)

ABSTRACT Arthrospira platensis is a source of proteins, polysaccharides, polyunsaturated fatty acids (in tiny amounts) and phenolic components that give it different biological properties. The type of nutrients present in the culture medium can influence the microalga metabolism and, consequently, the growth, composition and content of nutrients, bioactive compounds and biological properties. In this work, A. platensis was cultivated in the laboratory using six culture media (Zarrouk as a standard medium, and Media 1–5), containing different nitrogen sources, and subsequently harvested and dried by freeze-drying or cold-drying. Medium 4, with KNO3 as nitrogen source, proved to optimize the productivity of A. platensis (with a maximum yield of 1.34 ± 0.08 g l–1, better than the Zarrouk medium), to maximize protein synthesis (up to 70% w/w) and to increase the antioxidant activity. Conversely, Medium 3, containing KNO3 and NH4NO3, was the best to optimize carbohydrates and lipids (up to 30% w/w and 2% w/w, respectively). Both Media 3 and 4 were able to increase chlorophyll-a and carotenoid synthesis compared to the other tested media. Cold-drying has proved to be an excellent alternative to the more expensive lyophilization: the content of carotenoids and antioxidant components was better preserved. This research has shown that low-cost media and cold-drying can be a promising strategy for obtaining a high biological value product, usable in both the pharmaceutical and food sectors, reducing production times and costs.

Low sensitivity of conventional fungal agars in fungemia by Rhodotorula mucilaginosa: description of two cases

BackgroundAlthough most bloodstream yeast infections are caused by Candida spp., infections by rare or less common species have increased in recent years. Diagnosis of infections caused by these species is difficult due to the lack of specific symptoms and adequate diagnostic tools.Cases presentationWe describe two cases of fungemia by Rhodotorula mucilaginosa within a few months of each other, in a secondary Spanish hospital. In both cases, diagnosis was challenging. Blood subcultures in conventional fungal media were persistently negatives and the use of non-conventional fungal media was essential for isolating the yeasts and achieving a correct diagnosis. 1–3 beta-d-glucan detection and a panfungal PCR assay were helpful techniques to confirm the diagnosisConclusionIt is highly important to establish an early diagnosis for fungemia. The process is challenging because often non-specific symptoms are presents. When yeasts grow in blood cultures other genera than Candida spp. could be the cause of infection. Patient risk factors should be assessed to incorporate alternative culture media and the available rapid diagnostic test, in order to provide an early recognition of the pathogen.

Experimental Evaluation of Seaweeds Liquid Extracts as an Alternative Culture Medium on the Growth and Proximate Composition of Picochlorum maculatum

Experimental Evaluation of Seaweeds Liquid Extracts as an Alternative Culture Medium on the Growth and Proximate Composition of Picochlorum maculatum

GROWTH IN VITRO OF Schomburgkia crispa Lindley IN THE INCREASED CULTURE OF Salvia hispanica L.

O uso de meios de cultura alternativos para o cultivo in vitro de plantas está sendo cada vez mais utilizado para atender as necessidades nutricionais diminuindo os custos de produção. Neste contexto, o presente trabalho teve como objetivo avaliar a influência das concentrações dos sais do meio de cultura MS e do extrato de chia no crescimento in vitro de plântulas de Shamburquia crispa Lindley. O delineamento experimental utilizado foi inteiramente casualizado - DIC, em esquema fatorial 5x5, sendo os tratamentos compostos por diferentes concentrações de sais do meio de cultura MS (0, 25, 50, 75 e 100%) e concentrações do extrato de chia (0; 3,5; 7,0; 10,5 e 14,0 g L-1), com quatro repetições e quatro plântulas por parcela. Os dados foram submetidos à análise de variância e para a comparação das médias dos tratamentos, foi realizado o teste Scott Knott a 5% de significância. Todas as análises foram realizadas com o auxílio do programa SISVAR 5.3. Decorridos 90 dias da instalação, foram analisados: taxa de sobrevivência (%), número de folhas, número de raízes, comprimento da maior raiz e comprimento da parte aérea. A redução dos sais do meio de cultura MS (0, 25 e 50%) associada à adição do extrato de chia (10,5 e 14 g L-1) promovem maior crescimento e desenvolvimento de explantes de Schomburgkia crispa Lindley cultivadas in vitro.

Characterization of Arthrospira sp (Spirulina) Biomass Growth in Hydroponic Waste Solution: A Review

The use of Spirulina (Arthospira sp) as a food source has stood out for being a potential source of proteins, fatty acids, vitamins, and others. However, the cultivation medium for its production is relatively expensive. Then, an alternative culture medium is necessary for reducing costs. So, hydroponics drainage effluent, a solution rich in nutrients, represents an alternative source of substrate for microorganism cultivation. Thus, this work aimed to characterize the biomass of Arthospira sp grown in an alternative culture medium based on hydroponic effluent through a systematic review. We noted a trend to reduce biomass growth and bioactive compounds due to an excess of iron and copper in the effluent. The study concluded that it needs to supplement the residual medium with a synthetic medium for its use to obtain biomass for commercial utilization.

Enhancement of Biomass and Lipid Productivities of Scenedesmus sp. Cultivated in the Wastewater of the Dairy Industry

Microalgae are photoautotrophic microorganisms capable of producing compounds with potential bioenergetic applications as an alternative energy source due to the imminent exhaustion of fossil fuels, their impact on the environment, and the constant population increase. The mass cultivation of these microorganisms requires high concentrations of nutrients, which is not profitable if analytical grade culture media are used. A viable alternative is the use of agro-industrial wastewater, due to the metabolic flexibility of these microorganisms and their ability to take advantage of the nutrients present in these substrates. For the reasons mentioned above, the effect of the cultivation in wastewater from cheese processing on the growth parameters and biomass composition of Scenedesmus sp. was evaluated, and its nutrient removal capacity determined. A high lipid concentration was obtained in the cultures with the dairy effluent (507.81 ± 19.09 mg g−1) compared to the standard culture medium, while the growth parameters remained similar to the control medium. Scenedesmus sp. achieved high percentages of nutrient assimilation of the wastewater used (88.41% and 97.07% for nitrogen and phosphorus, respectively). With the results obtained, the feasibility of cultivating microalgae in agro-industrial wastewater as an alternative culture medium that induces the accumulation of compounds with potential bioenergetic applications was verified.

Production of arachidonic acid by Mortierella alpina using wastes from potato chips industry.

This research aimed to determine the potential use of wastes from the potato chips industry as a carbon source to develop an economical culture medium for the production of biomass, lipids and arachidonic acid (ARA) by Mortierella alpina. A synthetic culture medium was optimized using a Plackett-Burman and central composite rotatable design, and used as a base to evaluate and characterize the potential use of wastes from the potato chips industry as carbon sources for the production of biomass, lipids and ARA by M. alpina. The waste was selected among other solid and liquid hydrolysed residues/by-products, and local low-cost alternatives for nitrogen sources were also evaluated. After 6days of fermentation, the biomass concentration reached 20gl-1 with 40% of total lipids, and a 35% ARA content in the lipids fraction. Savings in production were calculated using a sensitivity analysis for the alternative culture medium in different scenarios. This study showed a 7% savings in culture media expenses in the production of ARA-enriched biomass of M. alpina, compared to the conventional synthetic culture medium, when waste from the potato chips industry was used as an alternative source of carbon and macro/microelements, supplemented with a low-cost yeast extract alternative. The demonstration of the use of potato chips wastes as a low-cost carbon source for the biomass, lipids and ARA production, suggesting an eco-friendly alternative for the use of agri-food wastes for valuable metabolites production.

Exploiting the use of agro-industrial residues from fruit and vegetables as alternative microalgae culture medium

There is a need for searching new microalgae species, and the most suitable strategy to increase the cost-effectiveness of a microalgae culture system is to use resources of low costs, such as residues. This study aimed to evaluate the cultivation of microalgae isolated from the Brazilian Northeast region (Lagerheimia longiseta, Monoraphidium contortum, and Scenedesmus quadricauda) in an alternative medium of low cost (biocompost of discarded fruits and vegetables) with a view to possible applications in the food industry. Microalgae cultivated in the conventional synthetic medium was used as control. The cultivation of microalgae in the alternative medium allowed suitable cell growth, and improved the antioxidant activity and the levels of monounsaturated fatty acid and polyunsaturated fatty acid compared to the synthetic medium. The cultivation of S. quadricauda and L. longiseta species in the alternative medium resulted in increased protein content and/or total phenolic content, and improved health indices (lower levels of atherogenic, thrombogenic, and hypercholesterolemic saturated fatty acids indices, and higher levels of desired fatty acids index) compared to cultivation in synthetic medium. The cultivation of M. contortum in the alternative medium contributed to the production of higher lipid content, mainly saturated fatty acid (palmitic acid), which contributed negatively to the health indices. This study proved that S. quadricauda and L. longiseta microalga species from freshwaters have significant potential for distinct applications in functional food industries, and the biocompost of discarded fruits and vegetables is a suitable medium for microalgae cultivation.

Technological variables influencing the growth and stability of a silage inoculant based on spray-dried lactic acid bacteria.

To investigate the influence of different culture media and fermentation conditions on growth performance and viability of three lactic acid bacteria (LAB) strains with potential as silage inoculants, and to optimize spray-drying in order to enhance survival to dehydration and storage stability. In house-formulated MRS was a suitable low-cost culture medium for Lactobacillus plantarum Hv75, Pediococcus acidilactici 3903 and L. buchneri B463. Uncontrolled pH biomass production conferred enhanced stability during storage at 4°C after spray-drying. The use of whey protein concentrate 35 (WPC)-maltodextrin (M) as matrix, inlet temperature of 145-150°C and air flow rate of 601lh-1 was adequate for the production of dehydrated LAB. According to the desirability function, at this optimized condition, moisture content, yield and solubility were predicted to be 3·96, 73·68 and 90·36% respectively. Those conditions also showed a decrease of 0·855 log CFU per gram after drying, no loss in viability at 4°C for 6months and 1 log CFU per gram reduction at 25°C. Stable and economically feasible dehydrated LAB cultures were obtained using alternative culture media, fermentation under uncontrolled pH and optimizing spray-drying process conditions through the desirability function method. Our results can be utilized for efficient production and commercialization of several dry LAB.

A review on Spirulina: alternative media for cultivation and nutritive value as an aquafeed

AbstractThe nutritional value of spirulina in aquafeeds has been studied extensively as a fishmeal replacer or as a functional feed additive to increase growth and health performance of fish in the last decade. Despite increased interest in spirulina research in aquaculture, its increased production cost still limits its use in commercial rations. Zarrouk’s medium is the standard substrate used for growing spirulina. Although this medium offers optimal biomass production, it incurs higher costs due to the expensive components needed for concocting the medium. In this regard, development of cost‐effective alternative culture medium or techniques is necessary for the industrial‐scale production of these microalgae. This paper reviews research on different alternative media for spirulina cultivation and its nutritive value as an aquafeed. Numerous alterations in the composition of Zarrouk’s medium with cheaper chemical ingredients have been explored as a potential substitute for cost‐effective microalgae cultivation. Industrial and processing wastes and by‐products with appropriate nutrient profiles for growing spirulina are also being considered as alternative culture media. The use of rice bran or other cheaper agricultural by‐products, as a cost‐effective growth medium for commercial‐scale spirulina production has yet to be explored extensively in agriculture‐based countries, especially in rice‐producing Asian countries.

Resíduos agroindustriais como insumos promissores para obtenção de bioprodutos por leveduras - uma breve revisão

As leveduras desempenham um papel fundamental na natureza e, dada a sua versatilidade bioquímica e fisiológica, muitas delas podem ter suas rotas metabólicas direcionadas e otimizadas para a biossíntese de produtos específicos. Frente a essa potencialidade, novas contribuições que se debrucem sobre o estudo de processos fermentativos envolvendo leveduras como protagonistas, em especial para a produção de produtos de valor agregado a partir de vegetais perdidos e seus resíduos, revestem-se de importância. Dentre esses produtos, cita-se particularmente, nesta breve revisão, a produção de enzimas pectinolíticas e o potencial probiótico de leveduras em meios de cultura alternativos, como uma maneira de oportunizar esses processos de forma econômica e ambientalmente viável.

Optimization of a culture medium based on forage palm for δ-endotoxin production

An alternative culture medium based on forage palm (Cactaceae) and inexpensive nitrogen sources for δ-endotoxin production by Bacillus thuringiensis var. berliner (Btb) was optimized using response surface methodology and the influence of the nitrogen sources on biological activity and morphological properties of the δ-endotoxin has been studied. Effects of forage palm extract concentration (15–25%), ammonium sulphate (0–1 g/L), ammonium chloride (0–1 g/L) and urea concentrations (0–1 g/L) on δ-endotoxin production were studied by a 2 4 full factorial design and optimized by a 2 2 central composite design (CCD). Statistical analysis showed that forage palm and ammonium sulphate were the main factors to enhance δ-endotoxin production. Different nitrogen sources influenced on biological activity and morphology of crystals. Culture medium constituted by forage palm extract and ammonium sulphate increased δ-endotoxins production in the form of bipyramidal crystals and was more toxic to Spodoptera frugiperda . Forage palm extract and ammonium sulphate were considered to optimization of δ-endotoxin production. Maximum δ-endotoxin production (356 mg/L) took place at 30.7% (w/v) of forage palm extract concentration and 1.016 g/L of ammonium sulphate. • δ-endotoxins were produced in an inexpensive forage palm-based culture medium. • N. cochenillifera is more appropriate as a carbon source to δ-endotoxin production. • Ammonium sulphate increases the δ-endotoxin production and its toxicity. • The optimum medium had 30.7% N. cochenillifera and 1.016 g/L ammonium sulphate. • Optimum culture medium allowed the production of 356 mg/L of δ-endotoxin.

Biosynthesis of vitamin B12 by Propionibacterium freudenreichii subsp. shermanii ATCC 13673 using liquid acid protein residue of soybean as culture medium.

Vitamin B12 deficiency still persists, mainly caused by low intake of animal food products affecting vegetarians, vegans, and populations of underdeveloped countries. In this study, we investigate the biosynthesis of vitamin B12 by potential probiotic bacterium using an agroindustry residue, the liquid acid protein residue of soybean (LAPRS), as a low-cost, animal derivate-free alternative culture medium. Cultures of Propionibacterium freudenreichii subsp. shermanii ATCC 13673 growing in LAPRS for vitamin B12 biosynthesis were studied using the Plackett-Burman experimental approach, followed by a central composite design 22 to optimize the concentration of significant variables. We also performed a proteolytic treatment of LAPRS and evaluated the optimized-hydrolyzed medium influence on the microbial growth and metabolism in shaker flask and bioreactor experiments. In this all-plant source medium, P. freudenreichii subsp. shermanii produced high concentrations of cells and high amounts of vitamin B12 (0.6 mg/g cells) after process optimization. These results suggest the possibility of producing vitamin B12 by a potential probiotic bacterium in a very cheap, animal derivate-free medium to address the needs of specific population groups, at the same time reducing the production costs of this essential vitamin.

Manejo de macrófitas proveniente de infestação em ambientes naturais como suplemento para produção da microalga de interesse econômico Scenedesmus acuminatus

Este estudo avaliou a influência de seis meios de cultura alternativos (um a base de NPK, dois contendo extrato de Pistia stratiotes e Eichhornia crassipes enriquecido de NPK, e outros três com diferentes diluições de extrato puro de E. crassipes, (Eic 40, Eic 85, Eic 130) em comparação ao meio de cultura comercial ASM-1, no desenvolvimento da microalga Scenedesmus acuminatus. Foram avaliados taxa de crescimento (K), densidade, volume celular, biovolume e qualidade nutricional dos meios de cultura. Os meios de culturas testados apresentaram concentração de nutrientes suficiente para o crescimento da microalga. As taxas de crescimento de ASM-1 (0,439) foram similares (p>0,05) a NPK, Pis + NPK e Eic + NPK. Eic 85 e Eic 130 apresentaram os valores mais baixos (0,201; 0,150, respectivamente) e similares entre si. Os volumes dos tratamentos não diferiram de ASM-1 (135,427 μm3), somente o Eic NPK e Eic 130 apresentaram valores menores (86,780 e 100,401 μm3, respectivamente). O maior biovolume e densidade celular foram encontrados em Eic NPK (1,273 mm3.L-1 e 1047 x 104 cél.mL-1). Os tratamentos apresentaram capacidade de nutrição muito similar ao ASM-1, podendo substitui-lo no cultivo de S. acuminatus, em especial Eic + NPK que apresentou maior densidade e biovolume. Os tratamentos com extrato puro, apesar de apresentarem resultados mais baixos, ainda assim possibilitaram o desenvolvimento da alga.

Production of Paper Using Bacterial Cellulose and Residue from the Sugar and Alcohol Industry

The production of a fibrous surface formed from a composite of sugarcane bagasse and bacterial cellulose was the objective of this research. For this process the bagasse was grinded and cleaned, followed by the addition of a chemical substance capable of separating the cellulose fiber from the natural binders. The vegetable (VC) fiber originated from sugarcane bagasse (SCR) was mixed with bacterial cellulose (BC) fiber, which was produced in an alternative culture medium that used maize maceration liquor as a carbon and nitrogen source. The celluloses were ground, mixed and sieved to dryness. The resulting dry composite of the mixture was characterized by FTIR, DRX and TGA. The biomaterial obtained from the combination of BC and VC showed the same quality as a dense and rigid paper, with the advantage of not requiring the addition of chemicals and avoiding the consumption of a large volume of water. Infrared confirms the same chemical groups for BC and VC. Crystallinity and degradation did not show a large change for crystallinity index and degradation temperature when SCR was incorporated into BC. The pulp production process does not harm the environment at any time, since there is no emission of pollutants. An environmentally friendly product was obtained from sugarcane bagasse, a byproduct of ethanol and sugar manufacturing in BC mills and nanofibrils, producing high quality fibers, high purity and biodegradability, which makes the paper 100% recyclable.Keywords: Bacterial Cellulose, Gluconacetobacter hansenii, Sugarcane Bagasse,

Isolation of a freshwater microalgae and its application for the treatment of wastewater and obtaining fatty acids from tilapia cultivation.

In the present study, isolated strains of the microalgae Chlamydomonas sp. (CH) and Chlorella vulgaris (CV) were used to treat aquaculture wastewater and to obtain fatty acids and from a fattening culture of tilapia. The microalgae were cultivated for 11days in tubular photobioreactors with an operating volume of 2L, constant aeration and illumination. High removal rates of NO3- and PO43- were achieved for both Chlamydomonas sp. (84.7% and 96%, respectively) and Chlorella vulgaris (94.6 and 97.9%, respectively). The maximum biomass productivity achieved by Chlamydomonas sp. was 0.06 and 0.10 gL-1d-1 for Chlorella vulgaris. Therefore, tilapia wastewater contained the necessary nutrient concentration for algal growth and development. Chlamydomonas sp. biomass lipid content was 69%, while that of Chlorella vulgaris was 40%. The lipid profile of both microalgae was abundant in palmitic acid (78% for Chlamydomonas sp. and 35% for Chlorella vulgaris). This fatty acid is suitable for biodiesel production. Tilapia wastewater is low-cost alternative culture medium as it contains the necessary nutrient concentration for microalgae development and growth.

Fibrinogen-Depleted Equine Platelet Lysate Affects the Characteristics and Functionality of Mesenchymal Stem Cells.

Fetal bovine serum (FBS) is widely used to culture mesenchymal stem cells (MSCs) in the laboratory; however, FBS has been linked to adverse immune-mediated reactions prompting the search for alternative cell culture medium. Platelet lysate (PL) as an FBS substitute has been shown to promote MSCs growth without compromising their functionality. Fibrinogen contained in PL has been shown to negatively impact the immune modulating properties of MSCs; therefore, we sought to deplete fibrinogen from PL and compare proliferation, viability, and immunomodulatory capacities of MSCs in FBS or PL without fibrinogen. We depleted fibrinogen from equine platelet lysate (ePL) and measured platelet-derived growth factor-beta (PDGF-β), transforming growth factor-beta (TGF-β) and tumor necrosis factor-alpha (TNF-α) through ELISA. First, we determined the ability of 10% ePL or fibrinogen-depleted lysate (fdePL) compared with 10% FBS to suppress monocyte activation by measuring TNF-α from culture supernatants. We then evaluated proliferation, viability, and immunomodulatory characteristics of bone marrow-derived MSCs (BM-MSCs) cultured in FBS or ePL with or without fibrinogen. Growth factor concentrations decreased in ePL after fibrinogen depletion. Lipopolysaccharide (LPS)-stimulated monocytes exposed to ePL and fdePL produced less TNF-α than LPS-stimulated monocytes in 10% FBS. BM-MSCs cultured in fdePL exhibited lower proliferation rates, but similar viability compared with BM-MSCs in ePL. BM-MSCs in fdePL did not effectively suppress TNF-α expression from LPS-stimulated monocytes compared with BM-MSCs in FBS. Depleting fibrinogen results in a lysate that suppresses TNF-α expression from LPS-stimulated monocytes, but that does not support proliferation and immune-modulatory capacity of BM-MSCs as effectively as nondepleted lysate.

Dinámica del crecimiento in vitro de Phytophthora cinnamomi en medios de cultivo alternativos

México es el principal país productor, consumidor y exportador de aguacate el en mundo. El estudio de la macrobiótica del suelo cobra mayor importancia en el complejo marchitez del aguacate debido que en esta patología pueden estar implicados agentes causales de tipo biótico y abiótico, siendo el Oomycete Phytophthora cinnamomi Rands la principal enfermedad radical del aguacate. El objetivo de la presente investigación fue evaluar la dinámica del crecimiento in vitro de Phytophthora cinnamomi en medios de cultivo alternativos. En los meses de octubre y noviembre de 2016, se recolectaron muestras de raíces en arboles de aguacate (Persea americana Mill. var. Hass) en la huerta experimental del INIFAP ubicada en San Juan Nuevo Parangaricutiro, Michoacán. Los aislados se identificaron morfológica y molecularmente. Se registró las medidas de diámetros de crecimiento de cada cepa in vitro en función al tiempo. Se efectúo una regresión lineal mediante modelos lineales generalizados (GLM) para obtener los parámetros de intercepto y pendiente, significancia de la regresión y el valor de R2 ajustada. Los análisis se realizaron con el programa estadístico R 3.4. De acuerdo con los resultados obtenidos, el medio de cultivo Centeno-agar fue el más eficiente para la proliferación de P. cinnamomi.

Producción de celulosa bacteriana por Komagataeibacter xylinus utilizando desperdicio de mango como fuente de carbono alternativa

Bacterial cellulose (BC) is a high value-added nano-structured biopolymer with important biomedical applications.  Its biosynthesis from waste carbon sources might modify BC structure and properties, thus Mango pulp waste (MPW) was evaluated as an alternative culture medium for its production by the bacteria Komagataeibacter xylinus. The use of MPW could also decrease BC cost since the use pure sugars for its obtaining is expensive. The effect of different nitrogen sources and buffer addition to MPW-medium on the polymer yield was also investigated. Using MPW and yeast extract as a nitrogen source, a production of 6.32 g/L of BC was obtained after 16 days of static culture.  BC was characterized by SEM, XRD, TGA, FTIR and Water holding capacity (WHC). Chemical structure and thermal degradation of BC produced from MPW were similar to those of BC obtained with pure sugars (350°C). Crystallinity index of BC produced in mango-based medium was lower (62.7 % vs. 77.2 %); WHC was higher (108.7 % vs.  88.7 %); and fiber diameter was smaller (98.8 nm vs. 50.6 nm).