Estrutura e dinâmica do fitoplânctone bacterioplâncton em cultivos de camarão no Rio Grande do Norte-Brasil: impacto sobre o ambiente natural

This work aimed to study the structure and dynamic of Phytoplankton and Bacterioplankton in a complete cycle of shrimp cultivation (Litopenaeus vannamei) and determine the environmental factors responsible for the structural changes of these communities. The study was realized in a saltwater shrimp farm (Macaíba, RN), between September/2005 and February/2006, and in a freshwater shrimp farm (Ceará Mirim, RN), between May/2007 and September 2007. The samplings were collected weekly in saltwater farm and every fifteen days in freshwater farm. Total phosphorus, chlorophyll a and environmental parameters (pH, dissolved oxygen, salinity, temperature, depth and water transparency) were measured. Qualitative and quantitative analysis of the phytoplankton and bacterioplankton were carried out. The Shannon-Wiener ecologic indexes of diversity and the Pielou equitability indexes were calculated to the phytoplankton. Bacterial density was determined by epifluorescence microscopy. The data were statistically analyzed by Pearson correlation and t-Test. Chlorophycea were predominat in salt water and in the captation/drainage point (24 to 99%). Diatoms had higher wealth. The species Choricystis minor had the highest occurrence (100%) and dominance (90-100%), thus showing its adaptation to the high temperatures, salinity and low water transparency conditions. Filamentous Cyanobacteria like Oscillatoria sp., Pseudoanabaena sp. and Phormidium sp. had constant levels. The negative correlation between chlorophycea and water transparency, and the positive correlation between chlorophyll a and salinity, showed that the phytoplankton was well adapted to the low transparency and to the high salinity. The bacterioplankton was negatively correlated with the total phosphorus and salinity. In freshwater, Cyanobacteria were predominant (>80%), presenting some producers of toxins species like Microcystis sp., Aphanizomenon sp., Cylindrospermopsis raciborskii e Anabaena circinalis. Cyanobacterial density and total phosphorus and chlorophyll a concentrations exceeded the maximum value allowed by legislation. The means of total phosphorus varied from 264 to 627 Wg.L-1 and the means of chlorophyll a oscillated between 22 and 182 Wg.L-1. The phytoplankton species were selected by low availability of the light, high pH, temperature and high availability of total phosphorus. The bacterioplankton showed high densities (5,13 x 107 to 8,50 x107 Bac.mL-1). The studied environments (ponds and rivers) presented a high level of trophic state based on the high concentrations of chlorophyll a and total phosphorus and cyanobacteria dominance. The composition of species in the ponds and rivers was similar, as well as high concentrations of total phosphorus and chlorophyll a, highlighting the pollution caused by the discharges of the farms in natural environment

Comportamento do camarão de água doce macrobrachium rosenbergii (De Man, 1879) em cultivos misto e monossexo

Understanding the behavioral activities of freshwater shrimp in captivity is of paramount importance for the appropriate management of the species. In Brazil, the shrimp Macrobrachium rosenbergii is currently the most widely used species in the freshwater shrimp culture due to its high potential for cultivation and good market acceptance. Thus, the present study aimed to describe and characterize the behavioral activities of M. rosenbergii in monosex and in mixed (male and female) (manuscript 1, 2 and 3) populations and the growth performance of this species in restrictive feeding conditions and in different feeding management (manuscript 4 and 5, respectively) . Juvenile and adult shrimps were collected from ponds of the Aquaculture Station - Unidade Especializada em Ciências Agrárias - Universidade Federal do Rio Grande do Norte (UFRN), Macaíba/RN and then transferred to the Laboratório de Estudos do Comportamento do Camarão LECC (Laboratory for Shrimp Behavioral Studies) of the Universidade Federal do Rio Grande do Norte (UFRN). For each treatment , eight aquaria of 250 L (50 cm x 50 cm x 100 cm) were used in a closed recirculating water system with artificial lighting, constant aeration , continuous filtration through a biochemical and biological filter (canister filter), and fine sand as substrate . The water quality was monitored daily. The lab consisted of two rooms with artificial lighting system , controlled by a timer with dark / light cycle of 12:12 h . In manuscript 1, the behavioral categories of the species were presented through an ethogram, which described 31 behaviors, subdivided into general and agonistic behaviors. Manuscript 2 compared the behavioral profile of shrimps in male and in female monosex and mixed populations over 24 hours in laboratory. In three types (mixed, male monosex and female monosex) of populations during the light and dark phases of the 24 hour cycle, the shrimps showed higher occurrence of cleaning behavior. Manuscript 3 examined the influence of the color of the shelter on the frequency of its use and behavioral activities of shrimp in mixed, in male monosex and in female monosex populations over 24 hours. We observed that the shrimp M. rosenbergii burrow more frequently during the light phase in male monosex and mixed populations; they also tend to choose the black shelters. Female monosex populations tend to use red and orange shelters. In manuscript 4, we evaluated in laboratory the behavioral activities and growth performance of juvenile shrimps under food restriction. We observed that a mild food restriction may be used since there is no loss concerning the growth of the animals; feeding management on alternate days , compared to daily management can be financially productive both reducing labor costs and reducing the amount of feed used . Manuscript 5 evaluated the behavior of shrimps in monosex and in mixed populations, as well as the latency of reach the food according to feed offer (tray or food dispersal) . Our results indicate that animals adjust to both types of feed offer food dispersal as much as tray, but they spend more time to reach the feed when it is offered in trays (feeders). Comparing culture types (mixed, male monosex and female monosex), the latency to reach the food was lower for female monosex population. The data obtained in this study demonstrate the importance of identifying different pressures and environmental stimuli on the behavioral responses of this species. This knowledge would support management improvement to optimize the levels of animals‟ welfare, resulting in a better zootecnical performance

Estrutura e dinâmica do fitoplânctone bacterioplâncton em cultivos de camarão no Rio Grande do Norte-Brasil: impacto sobre o ambiente natural

This work aimed to study the structure and dynamic of Phytoplankton and Bacterioplankton in a complete cycle of shrimp cultivation (Litopenaeus vannamei) and determine the environmental factors responsible for the structural changes of these communities. The study was realized in a saltwater shrimp farm (Macaíba, RN), between September/2005 and February/2006, and in a freshwater shrimp farm (Ceará Mirim, RN), between May/2007 and September 2007. The samplings were collected weekly in saltwater farm and every fifteen days in freshwater farm. Total phosphorus, chlorophyll a and environmental parameters (pH, dissolved oxygen, salinity, temperature, depth and water transparency) were measured. Qualitative and quantitative analysis of the phytoplankton and bacterioplankton were carried out. The Shannon-Wiener ecologic indexes of diversity and the Pielou equitability indexes were calculated to the phytoplankton. Bacterial density was determined by epifluorescence microscopy. The data were statistically analyzed by Pearson correlation and t-Test. Chlorophycea were predominat in salt water and in the captation/drainage point (24 to 99%). Diatoms had higher wealth. The species Choricystis minor had the highest occurrence (100%) and dominance (90-100%), thus showing its adaptation to the high temperatures, salinity and low water transparency conditions. Filamentous Cyanobacteria like Oscillatoria sp., Pseudoanabaena sp. and Phormidium sp. had constant levels. The negative correlation between chlorophycea and water transparency, and the positive correlation between chlorophyll a and salinity, showed that the phytoplankton was well adapted to the low transparency and to the high salinity. The bacterioplankton was negatively correlated with the total phosphorus and salinity. In freshwater, Cyanobacteria were predominant (>80%), presenting some producers of toxins species like Microcystis sp., Aphanizomenon sp., Cylindrospermopsis raciborskii e Anabaena circinalis. Cyanobacterial density and total phosphorus and chlorophyll a concentrations exceeded the maximum value allowed by legislation. The means of total phosphorus varied from 264 to 627 Wg.L-1 and the means of chlorophyll a oscillated between 22 and 182 Wg.L-1. The phytoplankton species were selected by low availability of the light, high pH, temperature and high availability of total phosphorus. The bacterioplankton showed high densities (5,13 x 107 to 8,50 x107 Bac.mL-1). The studied environments (ponds and rivers) presented a high level of trophic state based on the high concentrations of chlorophyll a and total phosphorus and cyanobacteria dominance. The composition of species in the ponds and rivers was similar, as well as high concentrations of total phosphorus and chlorophyll a, highlighting the pollution caused by the discharges of the farms in natural environment

Abundância e distribuição sazonal de espécies autóctones de camarões peneídeos no canal de abastecimento do sistema primar de aquicultura orgânica (Laguna de Guaraíras, Tibau do Sul, Rio Grande do Norte, Brasil)

Guaraíras lagoon, located in Tibau do Sul in the eastern littoral of Rio Grande do Norte (Brazil), presents a permanent connection to the sea, which guarantees the occurrence of a rich biodiversity, which includes the autochthonous shrimp species Litopenaeus schmitti, Farfantepenaeus subtilis and Farfantepenaeus brasiliensis. In spite of being subject to a strong human intervention in the last decade, mainly related to the installation of shrimp (Litopenaeus vannamei) farms, the lagoon is still scarcely studied. The present study aims at characterizing the populations of the three autochthonous penaeid shrimp species inhabiting Guaraíras, taking into consideration their abundance and seasonal distribution in the inflow channel of Primar System of Organic Aquaculture (Tibau do Sul, Rio Grande do Norte, Brazil). Twelve monthly samples were carried out from May 2005 to April 2006 with the aid of a circular cast net in the inflow channel, which is daily supplied with water from Guaraíras. Sampling months were grouped in trimesters according to the total pluviosity, thus comprising four trimesters. Water salinity was monitored twice a week and temperature values registered on a daily basis at noon, during the study period. The daily pluviosity data from the municipality of Tibau do Sul were supplied by Empresa de Pesquisa Agropecuária do Rio Grande do Norte (EMPARN). Collected shrimp were identified, weighted, measured and sexed. L. schmitti specimens (0.2 g to 17.8 g) were distributed in 1.3 g weight classes intervals. From the eighth sampling month (December 2005) onwards, males were classified into three categories, in accordance with the development of their petasm: (a) rudimentary petasm, (b) partially formed petasm, and (c) completely formed petasm. Among the ecological variables, rainfall showed the greatest dispersion (s.d.=187.74Rainfall and abundance of L. schmitti were negatively correlated (r = -0.85) whereas its abundance and water salinity were positively correlated (r = 0.63). Among 1,144 collected individuals, 1,127 were L. schmitti, 13 were F. subtilis and 4 were F. brasiliensis, which corresponded to 98.51%, 1.14% and 0.35% of the total of collected individuals. L. schmitti occurred in 100 % of all samples. Differently, the presence of F. subtilis and F. brasiliensis was restricted to 33% and 17% of the collected samples, respectively. The present study confirmed the occurrence of L. schmitti, F. brasiliensis and F. subtilis in Guaraíras. However, this lagoon seems to be primarily inhabited by juvenile Litopenaeus schmitti. The population of L. schmitti analysed showed a seasonal pattern of distribution. In general, in the months of high salinity and absence of rain, the number of individuals was higher than in the wet months. Further studies on the reproductive biology and ecology of L. schmitti, F. brasiliensis and F. subtilis may elucidate questions referring to the abundance, period, and phase of occurrence of these shrimp genera in Guaraíras. Finally, the risks associated to the establishment of L. vannamei in the lagoon provide a novel outlet for studies in this biotope

Potencialidades de cultivo de macroalgas no litoral do Rio Grande do Norte (NE, Brasil): uma perspectiva de integração de dados ambientais em sistemas de informação geográfica (SIG)

Since centuries ago, the Asians use seaweed as an important source of feeding and are their greatest world-wide consumers. The migration of these peoples for other countries, made the demand for seaweed to increase. This increasing demand prompted an industry with annual values of around US$ 6 billion. The algal biomass used for the industry is collected in natural reservoirs or cultivated. The market necessity for products of the seaweed base promotes an unsustainable exploration of the natural banks, compromising its associated biological balance. In this context, seaweed culture appears as a viable alternative to prevent the depletion of these natural supplies. Geographic Information Systems (GIS) provide space and produce information that can facilitate the evaluation of important physical and socio-economic characteristics for the planning of seaweed culture. This objective of this study is to identify potential coastal areas for seaweed culture in the state of Rio Grande do Norte, from the integration of social-environmental data in the SIG. In order to achieve this objective, a geo-referred database composed of geographical maps, nautical maps and orbital digital images was assembled; and a bank of attributes including physical and oceanographical variables (winds, chains, bathymetry, operational distance from the culture) and social and environmental factors (main income, experience with seaweed harvesting, demographic density, proximity of the sheltered coast and distance of the banks) was produced. In the modeling of the data, the integration of the space database with the bank of attributes for the attainment of the map of potentiality of seaweed culture was carried out. Of a total of 2,011 ha analyzed by the GIS for the culture of seaweed, around 34% or 682 ha were indicated as high potential, 55% or 1,101 ha as medium potential, and 11% or 228 ha as low potential. The good indices of potentiality obtained in the localities studied demonstrate that there are adequate conditions for the installation of seaweed culture in the state of Rio Grande do Norte

Efeito da adição de colesterol e ecdisona na produção in vitro do baculovírus spodoptera frugiperda MNPV

Among the pests that attack corn crop in Brazil, there is Spodoptera frugiperda (JE Smith, 1797) (Lepidoptera: Noctuidae), known as fall armyworm, which is the major corn pest. Due to genetic instability during serial passage of baculoviruses in insect cell culture, the viral bioinseticides in vitro production development is the greatest challenge for mass production of this bioproduct. Successive passages of virus using extracellular viruses (BVs), necessary during viral bioinseticides production scaling up, leads to the appearance of aberrant forms of virus, a process so called as "passage effect ". The main consequence of passage effect is the production of occlusion bodies (OB) decrease, preventing its production using in vitro process. In this study, it was carried out a serial passage of baculovirus Spodoptera frugiperda multiple nucleopolyhedrovirus, isolate 18, using Sf21 cells. A decrease in the production of occlusion bodies from 170 to 92 in the third to fourth passage was observed. A factorial experimental design (22) was employed to verify the influence of two input variables, concentration of the hormone 20 - hydroxyecdysone (CH) and cholesterol (CC) on the values of response variables (volumetric and the specific OB production) of the process, seeking to define the optimum operating ranges trying to reverse or minimize the passage effect. The result indicated a negative influence of the cholesterol addition and positive effect in the hormone supplementation which the optimum range found for the concentrations studied were 8 to 10μg/mL and 5 to 6.5 mg / mL, for cholesterol and hormone concentrations respectively. New experiments were performed with addition of hormone and cholesterol in order to check the influence of these additives on the OB production independently. While the best result obtained from the factorial experiment was 9.4 x 107 OB/mL and 128.4 specific OB/cell, with the addition of only 6μg/mL 20-hydroxyecdysone these concentrations increased to 1.9 x 108 OB/mL and 182.9 OB/cell for volumetric and specific OB production, respectively. This result confirms that the addition of the hormone 20-hydroxyecdysone enhances the SfMNPV in vitro production process performance using Sf21 cells

Influência do estresse nutricional programado na composição da microalga isochrysis galbana

Global warming due to Greenhouse Gases (GHG) emissions, especially CO2, has been identified as one of the major problems of the twenty-first century, considering the consequences that could represent to planet. Currently, biological processes have been mentioned as a possible solution, especially CO2 biofixation due to association microalgae growth. This strategy has been emphasized as in addition to CO2 mitigation, occurs the production of biomass rich in compounds of high added value. The Microalgae show high photosynthetic capacity and growth rate higher than the superior plants, doubling its biomass in one day. Its culture does not show seasons, they grow in salt water and do not require irrigation, herbicides or pesticides. The lipid content of these microorganisms, depending on the species, may range from 10 to 70% of its dry weight, reaching 90% under certain culture conditions. Studies indicate that the most effective method to promote increased production of lipids in microalgae is to induce stress by limiting nitrogen content in the culture medium. These evidences justify research continuing the production of biofuels from microalgae. In this paper, it was studied the strategy of increasing the production of lipids in microalgae I. galbana with programmed nutritional stress, due to nitrogen limitation. The physiological responses of microalgae, grown in f / 2 with different concentrations of nitrogen (N: P 15,0-control, N: 5,0 P and N: P 2,5) were monitored. During exponential phase, results showed invariability in the studied conditions. However the cultures subjected to stress in stationary phase, showed lower biomass yields. There was an increase of 32,5% in carbohydrate content and 87.68% in lipids content at N: P ratio of 5,0 and an average decrease of 65% in protein content at N: P ratios of 5, 0 and 2.5. There were no significant variations in ash content, independently of cultivation and growth phase. Despite the limitation of biomass production in cultures with N: P smaller ratios, the increase of lipid accumulation highest lipids yields were observed as compared to the control culture. Given the increased concentration of lipids associated to stress, this study suggests the use of microalgae Isochrysis galbana as an alternative raw material for biofuel production

Monitoramento em tempo real de processos fermentativos por espectroscopia no infravermelho próximo (NIRS)

A chemical process optimization and control is strongly correlated with the quantity of information can be obtained from the system. In biotechnological processes, where the transforming agent is a cell, many variables can interfere in the process, leading to changes in the microorganism metabolism and affecting the quantity and quality of final product. Therefore, the continuously monitoring of the variables that interfere in the bioprocess, is crucial to be able to act on certain variables of the system, keeping it under desirable operational conditions and control. In general, during a fermentation process, the analysis of important parameters such as substrate, product and cells concentration, is done off-line, requiring sampling, pretreatment and analytical procedures. Therefore, this steps require a significant run time and the use of high purity chemical reagents to be done. In order to implement a real time monitoring system for a benchtop bioreactor, these study was conducted in two steps: (i) The development of a software that presents a communication interface between bioreactor and computer based on data acquisition and process variables data recording, that are pH, temperature, dissolved oxygen, level, foam level, agitation frequency and the input setpoints of the operational parameters of the bioreactor control unit; (ii) The development of an analytical method using near-infrared spectroscopy (NIRS) in order to enable substrate, products and cells concentration monitoring during a fermentation process for ethanol production using the yeast Saccharomyces cerevisiae. Three fermentation runs were conducted (F1, F2 and F3) that were monitored by NIRS and subsequent sampling for analytical characterization. The data obtained were used for calibration and validation, where pre-treatments combined or not with smoothing filters were applied to spectrum data. The most satisfactory results were obtained when the calibration models were constructed from real samples of culture medium removed from the fermentation assays F1, F2 and F3, showing that the analytical method based on NIRS can be used as a fast and effective method to quantify cells, substrate and products concentration what enables the implementation of insitu real time monitoring of fermentation processes

Produção de enzimas por fungos em fermentação semi-sólida utilizando bagaço de coco e pedúnculo de caju como substratos

The production of enzymes by microorganisms using organic residues is important and it can be associated with several applications such as food and chemical industries and so on. The objective of this work is the production of CMCase, Xylanase, Avicelase and FPase enzymes by solid state fermentation (SSF) using as substrates: bagasse of coconut and dried cashew stem. The microorganisms employed are Penicillium chrysogenum and an isolated fungus from the coconut bark (Aspergillus fumigatus). Through the factorial design methodology and response surface analysis it was possible to study the influence of the humidity and pH. For Penicillium chrysogenum and the isolated fungus, the coconut bagasse was used as culture medium. In another fermentation, it was used the mixture of coconut bagasse and cashew stem. Fermentations were conducted using only the coconut bagasse as substrate in cultures with Penicillium chrysogenum fungus and the isolated one. A mixture with 50% of coconut and 50% of cashew stem was employed only for Penicillium chrysogenum fungus, the cultivation conditions were: 120 hours at 30 °C in BOD, changing humidity and pH values. In order to check the influence of the variables: humidity and pH, a 2 2 factorial experimental design was developed, and then two factors with two levels for each factor and three repetitions at the central point. The levels of the independent variables used in ascending order (-1, 0, +1), to humidity, 66%, 70.5% and 75% and pH 3, 5 and 7, respectively. The software STATISTICA TM (version 7.0, StatSoft, Inc.) was used to calculate the main effects of the variables and their interactions. The response surface methodology was used to optimize the conditions of the SSF. A chemical and a physic-chemical characterization of the coconut bagasse have determined the composition of cellulose (%) = 39.09; Hemicellulose (%) = 23.80, Total Lignin (%) = 36.22 and Pectin (%) = 1.64. To the characterization of cashew stem, the values were cellulose (g) = 15.91 Hemicellulose (%) = 16.77, Total Lignin (%) = 30.04 and Pectin (%) = 15.24. The results indicate the potential of the materials as substrate for semisolid fermentation enzyme production. The two microorganisms used are presented as good producers of cellulases. The results showed the potential of the fungus in the production of CMCase enzyme, with a maximum of 0.282 UI/mL and the Avicelase enzyme the maximum value ranged from 0.018 to 0.020 UI/ mL, using only coconut bagasse as substrate. The Penicillium chrysogenum fungus has showed the best results for CMCase = 0.294 UI/mL, FPase = 0.058 UI/mL, Avicelase = 0.010 UI/mL and Xylanase = 0.644 UI/ mL enzyme, using coconut bagasse and cashew stem as substrates. The Penicllium chrysogenum fungus showed enzymatic activities using only the coconut as substrate for CMCase = 0.233 UI/mL, FPase = 0.031 to 0.032 UI/ mL, Avicelase = 0.018 to 0.020 UI/mL and Xylanase = 0.735 UI/ mL. Thus, it can be concluded that the used organisms and substrates have offered potential for enzyme production processes in a semi-solid cultivation

Levantamento e avaliação de critérios para a ampliação de escala da produção de biossurfactantes utilizando melaço como substrato

Biosurfactants are amphiphilic molecules synthesized by microorganisms such as bacteria, yeast or filamented fungi cultivated in various carbon sources among sucrose and hydrocarbons. These molecules are composed by a hydrophilic and hydrophobic part. They operate mostly at interfaces of fluids of different polarities. Because of this characteristic, they are potentially employed in numerous industries, such as the textile, medical, cosmetics, food and mainly in the petrochemical ones. Therefore industry has interest in developing new biosurfactant production processes in high scale, in order to become them economically competitive when compared to synthetic biosurfactants. This work aims to evaluate the biosurfactant production applying a non-conventional substrate sugar cane molasses proceeding from the sugar industry thus reducing the production costs. The strain identified as AP029/GLIIA, isolated from oil wells in Rio Grande do Norte state and used in these experiments belongs to the culture collection of Antibiotics Department of UFPE. The fermentation were carried out using different conditions according to a factorial planning 24 with duplicate at center point, in which the studied factors were molasse concentration, nitrate concentration, agitation and aeration ratio. The experiments were performed in a shaker at 38ºC of temperature. Samples were withdrawn in regular periods of time of up to 72 hours of fermentation in order to analyze substrate consumption, cellular concentration, superficial tension, critical micelle dilution (CMD-1 e CMD-2) as well as extracelullar protein production. The results showed a production of 3,480 g/L of biomass, a reduction of 41% on superficial tension, 67% of substrate consumption and 0,2805 g/L of extracellular protein

Influência das condições de cultivo na produção de antígenos recombinantes de Leishmania i. chagasi utilizando Escherichia coli M15 cultivada em incubador rotativo e biorreator

Escherichia coli has been one of the most widely used hosts in recombinant protein production, in both laboratory and industrial scale since the advent of recombinant DNA technology. Despite the substantial progress of studies on the molecular biology and immunology of infections, there is currently no medication-based prophylaxis capable of preventing leishmaniasis. As such, there is a great need to identify specific antigens for the development of vaccines and diagnostic kits against visceral leishmaniasis. Thus, the primary goal of the present study is to assess the influence of cultivation conditions on the production of Leishmania chagasi antigens, carried out in a rotating incubator and bioreactor. To that end, several assays were conducted to evaluate the kinetic behavior of antigens (648, 503) of Leishmania. i. chagasi in two different compositions of media (2xTY, TB), with and without an inducer. In order to improve expression, assays were performed in a benchtop bioreactor using the best conditions obtained in a rotating incubator, in addition to assessing the influence of stirring speed. Results show that high complexity of the cultivation medium favored kinetic growth of clones (648, 503). However, in assays submitted to induction by IPTG, this elevated complexity did not promote the expression of recombinant proteins. Expression of antigens 648 and 503 exhibited behavior associated with growth and, in terms of location, proteins 648 and 503 are intracellularly stored. Lactose may be the most adequate inducer in protein expression, when considering factors, cost, toxicity and stability. Elevated stirring may increase cell growth in clone 53, although it may not result in high concentrations for the protein of interest. On the other hand, positive results were obtained for all recombinant clones (648, 503) tested, confirmed by the electrophoretic profile

Análise citogenética de células-tronco derivadas do subendotélio da veia umbilical humana

Mesenchymal stem cells (MSCs) are known as a population of multi-potential cells able to proliferate and differentiate into multiple mesodermal tissues including bone, cartilage, muscle, ligament, tendon, fat and stroma. Several applications of the study of EC can be emphasized the therapeutic techniques such as guided bone regeneration by implantation of EC in the affected site, without the need for bone grafts, using titanium as a vehicle. The process of cryopreservation is essential for the maintenance of cell cultures, since the cell line is frozen, it can be maintained in liquid nitrogen for an indefinite period and then thawed for therapeutic or experimental purposes. The aim of this study was to isolate a population of MSCs derived from the subendothelium of the umbilical vein human (MSCs-SUVH) to assess cytogenetic analysis by the possibility of appearance of chromosomal changes in two different situations: MSCs-SUVH regarding the process of cryopreservation and MSCs-SUVH grown on the surface of titanium. Flow cytometry analysis revealed that, this cell population was positive for the markers CD29, CD73 and CD90, but there was no expression of hematopoietic lineage markers, such as CD14, CD34 and CD45 and demonstrated capacity for osteogenic differentiation. The chromosomes obtained from the primary culture of MSCs-SUVH were analyzed by GTW banding technique, and results are described as guidelines to ISCN 2005. There was not the emergence of clonal chromosomal changes in the MSCs-SUVH in different situations analyzed. However one of the strings presented a balanced paracentric inversion, probably a cytogenetic constitutional alterations, which was present before and after the experimental situations that the MSCs-SUVH was submitted

Distribuição diária das atividades comportamentais dos camarões marinhos Farfantepenaeus subtilis (Pérez Farfante, 1967) e Litopenaeus schmitti (Burkenroad, 1938) em condições laboratoriais

Marine aquaculture has developed in the last decades all over the world, especially related to shrimp management. In Brazil, the introduction of the species Litopenaeus vannamei has contributed to the success of the activity, even if there are problems associated with the management of a exotic species, such as new diseases and ecological pressure on native species. It has been emphasized the need of research for developing new methodology that will allow native species management, being the most important Farfantepenaeus subtilis and Litopenaeus schmitti. Most knowledge obtained from research with those species has generally used a technical approach and mainly focused feeding process. There are no specific behavioral data on their activity pattern which should be of great importance for the use of native species on commercial culture farms. So, it was our objective to study and compare the daily distribution of behavioral activities of the marine shrimp species Litopenaeus schmitti and Farfantepenaeus subtilis. Forty animals of each species, 5 individuals per aquarium, were maintained in aquaria containing 200L of sea water under continuous aeration and filtration. They were marked individually and were observed by the instantaneous focal time sampling, along 10 continuous days, in 6 daily 15 min observation windows, every two hour. In each window, behaviors and location position of the animals in the aquarium were registered at 1 min intervals. Food was offered 3 times a day, representing 10% of each aquarium biomass. Aquaria were maintained in artificial photoperiod, 12hour light/l2 hour dark, 4 aquaria in light cycle equivalent to the environmental one (light from 06:00 to 17:59 h and dark from 18:00 to 05:59 h) and the other 4 in the reverse light cycle (light from 18:00 to 05:59 h and dark from 06:00 to 17:59 h) to allow sequential behavioral observation in both phases of the 24 hour cycle. There was a clear distinction between the distribution of behavioral activities of F. subtilis and L. schmitti in the two phases. The activity pattern of Farfantepenaeus subtilis demonstrates that species has prominently night habits and a burying pattern during the light cycle. Exploration, inactivity and swimming were the most common activities. The behavioral pattern of Litopenaeus schmitti indicates that species is active along both phases of the light cycle, and the most evident behaviors were exploration, inactivity and swimming

Cultivo de algas marinhas como desenvolvimento de comunidades costeiras

In northeastern Brazil, the extraction of agar has increased considerably in recent decades mainly using macroalgae of the genus Gracilaria. The pressure of the harvest of seaweed of this genus has compromised the sustainability of this natural resource. Given the current framework of decline in production of algae in the state of Rio Grande do Norte, it is necessary the deployment of crops that will assist in the development of coastal areas. This research aimed to develop studies on growth, productivity, efficiency and quality of agar of G. birdiae so as to provide subsidies for crops on commercial scale. The study was carried out in dry and rainy periods using rafts of cultivation. Algal biomass and the physical and chemical parameters of water were measured every fifteen days. In laboratory, the resistance and quality of agar were analyzed. The relative growth rate (TCR) was determined by the formula: RGR = ln (final weight / weight initial). 100 / interval time of sampling. During the study, environmental factors as salinity and temperature remained relatively constant (around 35 PSU and 28°C, respectively). The mean values of biomass ranged around 1952.67 ± 576g in the rainy period and 1925.67 ± 450g in the dry period, and they presented no significant variations. The maximum value of growth (TCR) was recorded in the dry season (7.45%.day-1), with an average over the study of 4.35%.day-1. The yield of agar ranged from 22% to 15%, and its resistance ranged from 850 to 650g. cm². The average obtained for the two periods was approximately 750 g cm². These results demonstrate the great potential of Gracilaria birdiae for mariculture can be used as an sustainable activity for coastal communities

Estratégias de Produção in vitro de Bioinseticida Viral: influências do Isolado, da Cinética e do Modo de operação

Societal concerns about environmental sustainability has lead to the development of ecologically-friendly alternatives to chemical insecticides for crop protection. One such alternative is biological pest control. In particular, baculoviruses are well suited as insect biopesticides due to their narrow host specificity and relative ease of propagation. In Brazil, the baculovirus Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) is the main biological control agent employed for the soybean pest, Anticarsia gemmatalis. This baculovirus biopesticide is currently produced using caterpillars, but increasing market demand for the product has encouraged the development of an in vitro manufacturing process, which can be scaled up to much higher virus productivities. In this study, three wild-type AgMNPV isolates (AgMNPV-2D, AgMNPV-MP2 and AgMNPV-MP5) and a recombinant form (vAgEGT-LacZ) were characterised in terms of occlusion body (OB) production and infection kinetics, to enable future optimisation of the in vitro production process. These viruses were propagated using a Spodoptera frugiperda (IPLB-SF21) insect cell line grown in shaker-flask batch cultures. Among the virus isolates tested, AgMNPV-MP5 was found to be the best producer, yielding (5.3±0.85)x108 OB/mL after 8 days post infection. The characterisation of vAgEGT-LacZ propagation in suspension cell cultures has not been previously reported in the literature; hence it became the main focus for this thesis. In particular, it was carried out a study on the effect of the multiplicity of infection (MOI) on OB production. Five successive batches were performed getting a final production (8.9±1.42)x1014 occlusion bodies, considering that production is related for a bioreactor with final volume of 10m3. A low MOI associated with a fed-batch process for vAgEGT-LacZ production was found to support a 3-fold higher OB yield when compared to the default batch process (1.8x107 and 5.3x107 OB/mL, respectively). This yield is competitive with regards to the production process.