RESUMO
The intensive use of the antihypertensive losartan potassium (LOS) has culminated in its high occurrence in aquatic environments. However, insufficient studies had investigated its effects in non-target organisms. In this study, ecotoxicity of LOS was assessed in aquatic organisms from distinct trophic levels (Desmodesmus subspicatus, Daphnia magna, and Astyanax altiparanae). Genotoxicity was assessed by the comet assay in D. magna and A. altiparanae, and biochemical biomarkers for the fish. LOS was more toxic to D. subspicatus (EC50(72h) = 27.93 mg L-1) than D. magna (EC50 = 303.69 mg L-1). Subsequently, this drug showed to induce more DNA damage in D. magna than A. altiparanae, when exposed to 2.5 mg L-1. No significant stress responses were observed by the fish biomarkers, suggesting that higher trophic levels organisms are more tolerant to LOS toxicity. LOS showed relatively low toxic potential for a short period of exposure, but with different patterns of toxicity for the organisms from distinct trophic levels, contributing to further risk assessment of LOS.
Assuntos
Anti-Hipertensivos/toxicidade , Losartan/toxicidade , Poluentes Químicos da Água/toxicidade , Acetilcolinesterase/metabolismo , Animais , Organismos Aquáticos/efeitos dos fármacos , Organismos Aquáticos/genética , Organismos Aquáticos/crescimento & desenvolvimento , Organismos Aquáticos/metabolismo , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Characidae/genética , Characidae/metabolismo , Clorofíceas/efeitos dos fármacos , Clorofíceas/crescimento & desenvolvimento , Ensaio Cometa , Daphnia/efeitos dos fármacos , Daphnia/genética , Cadeia Alimentar , Glutationa/metabolismo , Glutationa Transferase/metabolismo , Músculos/efeitos dos fármacos , Músculos/metabolismoRESUMO
In this work, the previously proposed Fibonacci-type photobioreactor is scaled up and evaluated to produce Dunaliella salina. First, the composition of the culture medium was optimized to achieve maximal productivity. Next, the Fibonacci-type reactor was scaled up to 1250 L maintaining high solar radiation interception capacity of this type of reactor. Finally, the performance of the reactor for the production of green cells of Dunaliella salina at the environmental conditions prevailing in the Atacama Desert was evaluated. Data demonstrated that the proposed photobioreactor allows the temperature, pH and dissolved oxygen concentration to be maintained within the optimal ranges recommended for the selected strain. Both better exposure to solar radiation and photonic flow dilution avoids the use of cooling systems to prevent overheating under outdoor conditions. The system allows up to 60% more solar radiation to be intercepted than does the horizontal surface, likewise, allowing to maintain the pH efficiently through CO2 injection and to keep the dissolved oxygen concentration in acceptable ranges, thanks to its adequate mass transfer capacity. The biomass concentration reached up to 0.96 g L-1, three times higher than that obtained in a raceway reactor under the same environmental conditions, whereas productivity was up to 0.12 g L-1 day (2.41 g m-2 day). Maximum specific outdoor growth rates reached up to 0.17 day-1. Undoubtedly, this technology scaled up constitutes a new type of photobioreactor for use at the industrial scale since it is capable of maximizing biomass productivity under high light conditions.
Assuntos
Biomassa , Clorofíceas/crescimento & desenvolvimento , FotobiorreatoresRESUMO
Growth and biological conditions of Messastrum gracile were evaluated to compare the effect of photoautotrophic and mixotrophic cultivation on the increase of biomass production and chemical conditions cultured in macrophyte and commercial culture media. The growth rate (k) of M. gracile was different in the culture media, higher in mixotrophic cultivation for Lemna minor culture medium, whilst to Eichhornia crassipes and NPK culture media were higher in photoautotrophic cultivation. Mean lipid contents in photoautotrophic cultivation were 8.2% biomass dry weight, whereas they reached 19% biomass dry weight in mixotrophic cultivation. Protein contents were below 48% biomass dry weight in photoautotrophic cultivation and 30% biomass dry weight in mixotrophic cultivation. Messastrum gracile cultured in macrophyte culture media (E. crassipes and L. minor) and NPK culture medium provided satisfactory results with regard to lipid and protein contents in mixotrophic and photoautotrophic cultivations, respectively. Lipid and protein contents in alternative media were higher or similar to the CHU12 commercial culture medium.(AU)
O crescimento e as condições biológicas da microalga Messastrum gracile foram avaliados para comparar o efeito do cultivo foto-autotrófico e mixotrófico na produção de biomassa e condições químicas em meios de cultura comercial e de macrófitas. A taxa de crescimento (k) de M. gracile foi diferente entre os meios de cultura, sendo maior no cultivo mixotrófico para o meio Lemna minor, enquanto para os meios Eichhornia crassipes e NPK foram maiores no cultivo foto-autotrófico. Os teores de lipídios no cultivo foto-autotrófico foram de 8,2% da biomassa seca, enquanto que no mixotrófico atingiram 19% da biomassa seca. Os teores de proteína em cultivo foto-autotrófico estiveram abaixo de 48% da biomassa seca e 30% de biomassa seca no cultivo mixotrófico. Messastrum gracile cultivada em meios de cultura de macrófitas (E. crassipes e L. minor) e NPK apresentaram resultados satisfatórios em relação aos teores de lipídeos e proteínas nos cultivos mixotróficos e foto-autotróficos, respectivamente. Os teores lipídicos e proteicos em meios alternativos foram maiores ou semelhantes ao meio comercial CHU12.(AU)
Assuntos
Clorofíceas/crescimento & desenvolvimento , Microalgas/crescimento & desenvolvimento , Processos AutotróficosRESUMO
Abstract Microalgae are potential sources of a wide range of bioproducts. It is essential to choose the proper microalgae strain and culture condition to achieve an efficient production. The production yield of carotenoids by Dunaliella salina under the stress-induced culture conditions of nitrogen deprivation and excessive light intensity was evaluated. Also, a survey at laboratorial scale of the growth kinetics under different culture conditions of photoperiod, aeration, and agitation was performed for the seven species of green microalgae Ankistrodesmus fusiformis, Chlamydocapsa bacillus, Desmodesmus brasiliensis, Kirchneriella lunaris, Pseudokirchneriella subcapitata, and Scenedesmus obliquus. As a result, aeration of atmospheric air is enough to improve the growth kinetics of the seven species studied. Production of carotenoids was enhanced under stress by excessive light intensity. Although D. salina does not grow effectively under nitrogen deprivation, this stress condition may be used to quickly stimulate carotenoid production once the culture reaches a high cellular population.
Assuntos
Carotenoides/biossíntese , Clorofíceas/crescimento & desenvolvimento , Clorofíceas/química , Estresse Fisiológico , Biotecnologia , Cinética , BiomassaRESUMO
Microalgae are photosynthetic microorganisms widely used for the production of highly valued compounds, and recently they have been shown to be promising as a system for the heterologous expression of proteins. Several transformation methods have been successfully developed, from which the Agrobacterium tumefaciens-mediated method remains the most promising. However, microalgae transformation efficiency by A. tumefaciens is shown to vary depending on several transformation conditions. The present study aimed to establish an efficient genetic transformation system in the green microalgae Dunaliella tertiolecta using the A. tumefaciens method. The parameters assessed were the infection medium, the concentration of the A. tumefaciens and co-culture time. As a preliminary screening, the expression of the gusA gene and the viability of transformed cells were evaluated and used to calculate a novel parameter called Transformation Efficiency Index (TEI). The statistical analysis of TEI values showed five treatments with the highest gusA gene expression. To ensure stable transformation, transformed colonies were cultured on selective medium using hygromycin B and the DNA of resistant colonies were extracted after five subcultures and molecularly analyzed by PCR. Results revealed that treatments which use solid infection medium, A. tumefaciens OD600â¯=â¯0.5 and co-culture times of 72â¯h exhibited the highest percentage of stable gusA expression. Overall, this study established an efficient, optimized A. tumefaciens-mediated genetic transformation of D. tertiolecta, which represents a relatively easy procedure with no expensive equipment required. This simple and efficient protocol opens the possibility for further genetic manipulation of this commercially-important microalgae for biotechnological applications.