Implementing stepwise solvent elution in sequential injection chromatography for fluorimetric determination of intracellular free amino acids in the microalgae Tetraselmis gracilis

The concept of sequential injection chromatography (SIC) was exploited to automate the fluorimetric determination of amino acids after pre-column derivatization with ophthaldialdehyde (OPA) in presence of 2-mercaptoethanol (2MCE) using a reverse phase monolithic C(18) stationary phase. The method is low-priced and based on five steps of isocratic elutions. The first step employs the mixture methanol: tetrahydrofuran: 10 mmol L(-1) phosphate buffer (pH 7.2) at the volumetric ratio of 8:1:91; the other steps use methanol: 10 mmol L-1 phosphate buffer (pH 7.2) at volumetric ratios of 20:80, 35:65, SO:SO and 65:35. At a flow rate of 10 mu L s(-1) a 25 mm long-column was able to separate aspartic acid (Asp), glutamic acid (Glu), asparagine (Asn), serine (Ser), glutamine (Gln), glycine (Gly), threonine (Thr), citruline (Ctr), arginine (Arg), alanine (Ala), tyrosine (Tyr), phenylalanine (Phe), ornithine (Orn) and lysine (Lys) with resolution >1.2 as well as methionine (Met) and valine (Val) with resolution of 0.6. Under these conditions isoleucine (Ile) and leucine (Leu) co-eluted. The entire cycle of amino acids derivatization, chromatographic separation and column conditioning at the end of separation lasted 25 min. At a flow rate of 40 mu L s(-1) such time was reduced to 10 min at the cost of resolution worsening for the pairs Ctr/Arg and Orn/Lys. The detection limits varied from 0.092 mu mol L(-1) for Tyr to 0.51 mu mol L(-1) for Orn. The method was successfully applied to the determination of intracellular free amino acids in the green alga Tetraselmis gracilis during a period of seven days of cultivation. Samples spiked with known amounts of amino acids resulted in recoveries between 94 and 112%. (C) 2008 Elsevier B.V. All rights reserved.

Large scale laboratory cultures of Ankistrodesmus gracilis (Reisch) Korsikov (Chlorophyta) and Diaphanosoma biergei Korinek, 1981 (Cladocera)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cultivo de Ankistrodesmus gracilis (Reisch) Korsikov (Chlorophyta) em laboratório utilizando meio CHU12 e de macrófita com NPK

Two different culture media, namely CHU12 and inorganic fertilizer NPK (20-5-20) associated with macrophyte (Eichhornia crassipes) extract, were used to evaluate the development of A. gracilis. Growth rate, development, nutritional value and medium water quality were analyzed. A. gracilis in macrophyte+NPK medium had mean cell density (333x10(5) cells/mL) higher than medium CHU12 (302x10(5) cells/mL). Protein rate (12.52% PS), dry weight (397x10(7) pg/cell) and ashes (4.08x10(7) pg/cell) in A. gracilis was higher (p<0.05) than medium macrophyte+NPK. on the other hand, lipids, carbohydrates and fibers had the same rate (p>0.05) in both media. When hydrological variables in the culture medium of A. gracilis are taken into account, dissolved oxygen and free CO2 alone failed to have any significant difference (p>0.05) between the media employed. A. gracilis in the macrophyte+NPK medium had the same nutritional rate as the commercial medium CHU12, with excellent results for alga growth. Macrophyte medium may be used for large scale alga culture in the feed of fish larvae.

Carotenoid production by Rhodotorula rubra cultivated in sugarcane juice, molasses, and syrup

A produção de biomassa e de carotenoides por Rhodotorula rubra foi estudada em meios à base de caldo, melaço e xarope de cana. Avaliou-se o efeito da suplementação dos meios com nitrogênio na forma de ureia ou do nutriente comercial Nitrofos KL. O delineamento experimental utilizado foi o inteiramente casualizado, no esquema fatorial 3 × 3, sendo um dos fatores o substrato (caldo, melaço e xarope) e o outro a suplementação (controle, ureia e Nitrofos KL). Os resultados foram submetidos à análise de variância e teste de Tukey a 5% de probabilidade. As maiores produções de massa seca de levedura foram obtidas no meio à base de melaço suplementado com ureia ou Nitrofos KL (15,09 e 14,87 g/L, respectivamente). A produção de carotenoides intracelular foi elevada em todos os meios estudados sem suplementação (0,329 mg/g). Para a produção volumétrica, o melhor meio foi o melaço (2,74 mg/L), enquanto a suplementação com ureia e com Nitrofos KL produziu 2,55 e 2,32 mg/L, respectivamente. Os principais carotenoides produzidos foram toruleno, torularrodina e β-caroteno. No meio à base de caldo de cana sem suplementação, houve o menor consumo de carboidratos, enquanto que o meio com suplementação à base de ureia obteve o maior consumo.

Cyclic-loading of the human gracilis and semitendinosus muscle tendons: Study of young adult cadavers

During knee ligament reconstruction, the tendon graft is tensioned to prevent the occurrence of excessive graft elongation during the postoperative period. Tensioning may be achieved by applying a cyclic or static load to the graft during fixation. Although this procedure is part of the surgery, there is no consensus in international literature regarding ideal tension levels to be used in this procedure. This study was conducted on 10 tendons of the human gracilis muscle and 10 tendons of semitendinosus muscle removed from five male cadavers whose mean age was 20.8 years. These tendons underwent 10 in vitro strain cycles at three levels of deformation (2.5, 3, and 4%) and the value of the deforming load used for each cycle was recorded. The statistical analysis demonstrated that in order to attain the same level of deformation during the 10 cycles there was a reduction in the value of strain applied to the graft, observed at the three levels of deformation. It was concluded that the semitendinosus tendon presents a more uniform mechanical behavior and that there is a need for new graft tensioning protocols that consider the force associated with deformation.

ZOEAL MORPHOLOGY OF PACHYGRAPSUS-GRACILIS (SAUSSURE, 1858) (DECAPODA, GRAPSIDAE) REARED IN THE LABORATORY

The larval development of Pachygrapsus gracilis was studied in the laboratory under conditions of 25-degrees-C temperature and 20, 24, 28, 32 and 34 parts per thousand salinity. The objective of the study was to characterize the plankton phase of life of this species.

Utilização de meios alternativos e diferentes tipos de recipientes no cultivo de Ankistrodesmus gracilis (Reinsch) Korshikov (Chlorophyceae) em laboratório

A laboratory culture of Ankistrodesmus gracilis algae was evaluated by studying the biology of the species and its chemical composition in a traditional medium (CHU 12) and in two alternative culture media, NPK (20-5-20) and macrophyte (Eichhornia crassipes) + NPK, in three different types of recipients (fiberglass, carboy and plastic bag). First peak in the growth curve of Ankistrodesmus gracilis occurred on the ninth day in macrophyte + NPK medium (74.16 x 10 5 cells mL -1) in a fiberglass recipient. However, highest density (p < 0.01) was reported in medium CHU 12 (122.87 x 10 5 cells mL -1) in a plastic bag on the twelfth day. Cell density was over 70 x 10 5 cells mL -1 starting on the twelfth day. Growth rate of A. gracilis was similar (p > 0.05) in culture media in the three recipients. Protein and fiber were similar (p > 0.05) in the treatments, but lipids were higher (p < 0.05) in NPK. Nitrate, ammonia, total phosphorus and orthophosphate contents were over 1 mg L -1 in NPK (p < 0.01). Results show that alternative media, such as NPK and macrophyte + NPK, are possible for large-scale culture of A. gracilis cultured in three types of recipients. Costs are low, occupying less space when cultured in plastic bags and in the laboratory.

Uso de meio a base de esterco suíno no cultivo de Ankistrodesmus gracilis (Chlorophyta) em laboratório

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Morfoanatomia do fruto, semente e plântulas de Acalypha gracilis (Spreng.) Müll. Arg., Euphorbia cotinifolia L. e Jatropha gossypiifolia L. (Euphorbiaceae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Redescription of megalopa and juvenile development of Pachygrapsus gracilis (Decapoda: Grapsidae) from the Amazon region, reared in the laboratory

ABSTRACT: This present study re-describes the megalopa stage and provides detailed morphological descriptions and main growth changes observed in stages I through VII of the juvenile instars of the dark shore crab Pachygrapsus gracilis (Saussure, 1858), from the Amazon region. The specimens in this study were reared in the laboratory and the megalopae were collected at Ajuruteua beach in northeastern Pará, Brazil. Previous studies had described the megalopa of P. gracilis from Mexican waters, as well as those of Pachygrapsus transversus (Gibbes, 1850) and Pachygrapsus marmoratus (Fabricius, 1787). A comparison between the Mexican and Amazonian populations of P. gracilis revealed significant morphological differences. The main difference is the presence of 3 elongated setae on the 5th pereiopods of individuals of the Amazonian population. The setal number and their arrangement in the appendages also differed. In P. gracilis, the male and female genital openings are observed from the juvenile instar III, whereas differentiation in male pleopods is observed only in juvenile instar V. In females, the pleopods undergo rapid differentiation during juvenile instar VI. These morphological comparisons and other observations on development are briefly compared and discussed with reports for other species.

Desempenho de Ankistrodesmus gracilis (Reischi) Korsikov (Chlorophyta) cultivada em três meios de cultura em laboratório

Pós-graduação em Aquicultura - FCAV

IS CLEISTOSPERMY AND COENOSPERMY RELATED TO SPERM TRANSFER? A COMPARATIVE STUDY OF THE MALE REPRODUCTIVE SYSTEM OF PACHYGRAPSUS TRANSVERSUS AND PACHYGRAPSUS GRACILIS (BRACHYURA: GRAPSIDAE)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)