Effects of water regulation on biosynthesis of calycosin-7-O-β-D-glucoside in Astragalus membranaceus var. mongholicus / 中国中药杂志

The effects of water regulation on the biosynthesis of calycosin-7-O-β-D-glucoside in 2-year-old Astragalus membranaceus var. mongholicus were studied,and the mechanism was explained from the aspects of key enzyme gene expression and antioxidant enzyme system. The content of calycosin-7-O-β-D-glucoside was determined by HPLC,and the expression levels of six key enzyme genes( PAL,4 CL,CHS,CHI,IFS,13'H) in the synthesis pathway were analyzed by q RT-PCR. The activities of protective enzymes and contents of osmoregulation substances and malondialdehyde were also determined. In the water deficit group,the maximum concentration of calycosin-7-O-β-D-glucoside was 0. 49 mg·g-1 on the 24 th day of treatment. In the whole water regulation,the water deficit group outweighed the water adequate group in osmoregulation substance and MDA contents. The activities of A. membranaceus var.mongholicus antioxidant enzymes SOD,POD,and CAT increased during the initial period of water regulation,but decreased with time.The expression of PAL,CHS,and 13'H in the water deficit group was at a low level,and the 4 CL had active expression,slightly lower than that in the water adequate group. The expression of CHI and IFS elevated rapidly when water deficit occurred. Correlation analysis showed that the content of calycosin-7-O-β-D-glucoside was positively correlated with CHI expression( P<0. 01) and IFS expression( P<0. 05). Therefore,water regulation can change the accumulation pattern of calycosin-7-O-β-D-glucoside,and water deficit may be an effective way to increase its content. CHI and IFS are the key genes in response to water deficit.

Saline water and potassium fertilization in cultivation of grafted west indian cherry ˜BRS 366 Jaburu / Águas salinas e adubação potássica no cultivo de aceroleira 'brs 366 jaburu' enxertada

Agricultural exploitation in the semi-arid region of Northeast Brazil depends on the use of irrigation to guarantee safe production of crops. Nevertheless, the waters commonly used in this region have high levels of salts and require management strategies that make their use possible in agriculture. In this context, the present study aimed to evaluate water relations, photosynthetic pigments and growth of grafted West Indian cherry as a function of saline water irrigation and potassium (K) fertilization. The study was conducted under greenhouse conditions in lysimeters filled with eutrophic Regolithic Neosol with sandy loam texture, in the municipality of Campina Grande-PB, Brazil. Treatments were distributed in randomized blocks and consisted of two factors: two levels of irrigation water electrical conductivity ­ ECw (0.8 and 3.8 dS m-1) and four K doses (50, 75; 100 and 125% of the recommendation), with three replicates and one plant per plot. The 100% dose corresponded to 19.8 g of K2O per plant per year. The West Indian cherry crop was sensitive to water salinity of 3.8 dS m-1 in the post-grafting phase, resulting in a decline in photosynthetic pigment content and growth. Increasing K doses reduced the percentage of cell membrane damage and promoted increase in the synthesis of chlorophyll a and carotenoids in West Indian cherry plants. There was significant interaction between salinity levels and K doses for the leaf osmotic potential, water saturation deficit, percentage of cell membrane damage and chlorophyll b content in West Indian cherry plants.

A exploração agrícola na região semiárida do Nordeste brasileiro depende do uso da irrigação para garantir a produção das culturas com segurança; entretanto, nesta região, as águas utilizadas comumente possuem níveis elevados de sais e necessitam de estratégias de manejo que possibilitem sua utilização na agricultura. Neste contexto, objetivou-se com este trabalho avaliar as relações hídricas, os pigmentos fotossintéticos e o crescimento da aceroleira enxertada em função da irrigação com águas salinas e adubação potássica. A pesquisa foi realizada em condição de casa de vegetação em lisímetros preenchidos com um Neossolo Regolítico Eutrófico de textura franco-arenosa, no município de Campina Grande-PB. Os tratamentos foram distribuídos em blocos casualizados, e consistiram de dois fatores, sendo dois níveis de condutividade elétrica da água de irrigação - CEa (0,8 e 3,8 dS m-1) e quatro doses de potássio (50, 75; 100 e 125% da recomendação), com três repetições e uma planta por parcela. A dose de 100% correspondeu a 33,0 g de K2O por planta por ano. A cultura da aceroleira é sensível a salinidade da água de 3,8 dS m-1, na fase pós enxertia, resultando-se em declínio nos teores de pigmentos fotossintéticos e crescimento. Doses crescentes de potássio diminuíram o percentual de dano na membrana celular e promoveram aumento na síntese de clorofila a e carotenoides nas plantas de aceroleira. Houve interação significativa entre os fatores níveis salinos e doses de potássio para potencial osmótico foliar, déficit de saturação hídrica, percentual de dano na membrana celular e teor de clorofila b da aceroleira.

Respuesta fisiológica y molecular de Anadara tuberculosa (Arcoida: Arcidae) al estrés de salinidad / Physiological and molecular response of Anadara tuberculosa (Arcoida: Arcidae) to salinity stress

Resumen: La concha negra Anadara tuberculosa es una especie emblemática del ecosistema manglar que está actualmente en condición vulnerable. El desarrollo de su acuicultura requiere identificar biomarcadores moleculares, en particular asociados al estrés por salinidad en mira al inicio de programas de mejoramiento genético. Se recolectaron ejemplares de Anadara tuberculosa del manglar colindante a la Bahía de Puerto Pizarro (Tumbes, Perú) entre enero 2015 y febrero 2016. Estos individuos fueron sometidos a condiciones de estrés hipo-osmótico (extremo: 5 y 10 ppt); (moderado: 15 y 25 ppt) y sin estrés (grupo control: 33 ppt) por 16 días después de haber sido separados en grupos de diez animales y por triplicado. La presencia de biomarcadores del estrés por salinidad fue evaluada a nivel genético con la detección por PCR de 19 genes reportados como actores claves de la osmorregulación en bivalvos como ostras y mejillones y a nivel proteomico con la secuenciación de péptidos expresados en tejidos de animales expuestos a diferentes salinidades por espectrometría de doble masa. Ninguno de los marcadores genéticos probados pudo ser amplificado por PCR lo que sugiere que A. tuberculosa presente diferencias genéticas significativas en comparación con otros moluscos. El análisis proteómico realizado por MALDI TOF/TOF a nivel de tejido branquial de A. tuberculosa permitió identificar 26 péptidos expresados de formas presenciales y diferenciales a las diferentes salinidades evaluadas, resaltando posibles marcadores como la HSP70 y una proteína transmembrana de transporte de cloruro que están relacionadas con la adaptación a la salinidad. Estas secuencias aminoacídicas permitirán diseñar iniciadores nucleotidicos específicos a A. tuberculosa para la puesta en marcha de futuras investigaciones en ecofisiología de este importante recurso.

Abstract: The pustulose ark A. tuberculosa is an emblematic species of mangrove ecosystem that is currently in a vulnerable condition. The development of its aquaculture, to begin with genetic breeding programs, requires the identification of molecular biomarkers, particularly those associated with salinity stress. With this purpose, specimens of A. tuberculosa were collected from the adjacent mangroves of Puerto Pizarro bay (Tumbes, Perú), from January 2015 to February 2016. Different assays (groups of ten animals in triplicate) were undertaken in separated periods of 16 days: hypo-osmotic stress (extreme: 5, 10 ppt); (Moderate: 15, 25 ppt) and no stress (control group: 33 ppt). The presence of salinity stress biomarkers was assessed at the genetic level throughout PCR detection of 19 genes reported to be key actors in osmoregulation, and at the proteomic level with the sequencing of peptides (tandem mass spectrometry MALDI TOF/TOF), expressed in ark tissues exposed to different salinities. None of the tested genetic markers could be amplified by PCR, suggesting that A. tuberculosa has significant genetic differences compared to other mollusks. Proteomic analysis by mass spectrometry on A. tuberculosa gill tissue, allowed to identify 26 peptides expressed in presential and differential forms at different salinities, highlighting possible markers such as HSP70 and trans-membrane chloride channel transportation protein, to be related with salinity adaptation. These amino acid sequences will allow the design of target specific primers for A. tuberculosa, to implement future research in ecophysiology of this important fishery resource. Rev. Biol. Trop. 65 (3): 1142-1151. Epub 2017 September 01.

Acclimation of juvenile Mugil liza Valenciennes, 1836 (Mugiliformes: Mugilidae) to different environmental salinities

Survival and physiological parameters associated with metabolism and osmoregulation were evaluated in juveniles of the Lebranche mullet Mugil liza acclimated to different water salinities (5, 10, 20, 30, and 40‰) for 15 days. Room temperature (25ºC) and photoperiod (12L:12D) were fixed. Fish were fed twice-a-day with commercial diet (28% crude protein) until satiation. After acclimation, whole body oxygen consumption was measured and fish were euthanized and sampled for blood, gills, and liver. Whole body oxygen consumption and plasma osmolality did not change in the range of salinities tested. The isosmotic point was estimated as 412.7 mOsmol kg-1 (13.5‰). Gill Na+,K+-ATPase activity tended to be lower at 20 and 30‰, while liver glycogen content was significantly higher at 20‰ than at 5 and 40‰. These results indicate that juvenile M. lizais able to acclimate for a short-period of time (15 days) to a wide range of salinities (5-40‰). This condition is achieved through adjustments in gill Na+,K+-ATPase activity and carbohydrate metabolism to regulate plasma osmolality and aerobic/energy metabolism. Therefore, our findings support the idea of catching juveniles M. lizain sea water and rear them in estuarine and marine waters.

A sobrevivência e parâmetros fisiológicos associados ao metabolismo e a osmorregulação foram avaliados em juvenis da tainha Mugil lizaaclimatada à diferentes salinidades (5, 10, 20, 30 e 40‰) por 15 dias. Foram fixadas a temperatura (25ºC) e o fotoperíodo (12L:12D) da sala onde os experimentos foram realizados. Os peixes foram alimentados duas vezes ao dia com ração comercial (28% de proteína bruta) até a saciedade. Após aclimatação, foi medido o consumo corporal de oxigênio e os peixes foram eutanasiados e foram coletadas amostras de sangue, brânquias e fígado. O consumo corporal de oxigênio e a osmolalidade plasmática não variaram na faixa de salinidade testada. O ponto isosmótico foi estimado em 412,7 mOsmol kg-1 (13,5‰). A atividade da Na+,K+-ATPase branquial tendeu a ser menor em 20 e 30‰, enquanto o conteúdo de glicogênio hepático foi significativamente maior em 20‰ do que em 5 e 40‰. Estes resultados indicam que o juvenil de M. liza é capaz de se aclimatar a uma ampla faixa de salinidade (5-40‰) por um curto período de tempo (15 dias). Esta condição é atingida através de ajustes na atividade da Na+,K+-ATPase branquial e no metabolismo de carboidratos para regular a osmolalidade plasmática e o metabolismo aeróbico/energético. Portanto, nossos achados suportam a ideia de que é possível capturar juvenis da tainha M. liza em água do mar e cultivá-los em águas estuarinas e marinhas.

Expression studies on NA+/K+-ATPase in gills of Penaeus monodon (Fabricius) acclimated to different salinities.

The decapod crustacean Penaeus monodon survives large fluctuations in salinity through osmoregulation in which Na+/K+-ATPase (NKA) activity in the gills plays a central role. Adult P. monodon specimens were gradually acclimatized to 5, 25 and 35‰ salinities and maintained for 20 days to observe long term alterations in NKA expression. Specific NKA activity assayed in gill tissues was found to be 3 folds higher at 5‰ compared to 25‰ (isosmotic salinity) and 0.48 folds lower at 35‰. The enzyme was immunolocalized in gills using mouse α-5 monoclonal antibody that cross reacts with P. monodon NKA α-subunit. At 5‰ the immunopositive cells were distributed on lamellar tips and basal lamellar epithelium of the secondary gill filaments and their number was visibly higher. At both 25‰ and 35‰ NKA positive cells were observed in the inter-lamellar region but the expression was more pronounced at 25‰. Gill architecture was normal at all salinities. However, the 1.5 fold increase in NKA α-subunit mRNA at 5‰ measured by quantitative RT-PCR (qRT-PCR) using EF1α as reference gene was not statistically significant. The study confirms the osmoregulating ability of P. monodon like other crustaceans at lower salinities. It is likely that significant increase in NKA transcript level happens at an earlier time point. At higher salinities all three methods record only marginal or no change from isosmotic controls confirming the hypothesis that the animal largely osmoconforms in hyperosmotic environment.

The effect of salinity on osmoregulation and development of the juvenile fat snook, Centropomus parallelus (POEY) / O efeito da salinidade sobre a osmoregulacao e o desenvolvimento do juvenil de robalo-peva, Centropomus parallelus (POEY)

Eurihaline fish support waters with different salt concentration. However, numerous studies have shown that salinity can affect fish development. Thus, the effect of salinity change from 20 to 5 and 35 on survival, weight, length, gill chloride cell ultrastructure and gill Na+, K+ ATPase activity was evaluated in Centropomus parallelus following short-term (6, 24 and 96 hours) and long-term exposure (30 and 60 days). Salinity did not affect C. parallelus survival, final weight and length. The quantity of chloride cells increased visibly at salinities of 5 and 35, with the cells exhibiting the typical features of uptake and secretory cells, respectively. Na+, K+ ATPase activity in the gill of the C. parallelus was significantly greater at a salinity of 5 than at a salinity of 20 or 35 after 96 hours, but not after 30 or 60 days. These results indicate that salinity change from high to low salt water induces gill chloride cell and Na+, K+ ATPase activity adaptations after short-term exposure. However, after long-term exposure at salinity 5, gill Na+, K+ ATPase activity is no more necessary at high levels. The increase in salinity to 35 does not induce significant change in gills. Juveniles of C. parallelus may thus be capable of acclimating to salinities of 5 to 35 for 60 days without significant effects on development.

Peixes eurihalinos suportam águas com diferentes concentrações de sal. Contudo, muitos estudos têm mostrado que a salinidade pode afetar o desenvolvimento do peixe. Portanto, o efeito da mudança de salinidade de 20 para 5 e 35 na taxa de sobrevivência, peso, comprimento, morfologia das células de cloreto branquiais e atividade da Na+, K+ ATPase foram avaliadas no Centropomus parallelus após curto (6, 24 e 96 horas) e longo tempo de exposição (30 e 60 dias). A salinidade não afetou a sobrevivência, o peso e comprimento final do robalo-peva. A quantidade de células de cloreto aumentou visivelmente nas salinidades 5 e 35, exibindo morfologias típicas de células que absorvem e secretam sal, respectivamente. A atividade da Na+, K+ ATPase nas brânquias do C. parallelus foi significativamente maior na salinidade 5 do que nas salinidades 20 ou 35 após 96 horas, mas não após 30 e 60 dias. Esses resultados indicam que a mudança de alta para baixa salinidade provoca adaptações nas células de cloreto e na atividade da Na+, K+ ATPase branquial em curto prazo. Contudo, após longa exposição na salinidade 5, a alta atividade da Na+, K+ ATPase branquial não é mais necessária. O aumento de salinidade para 35 não induz mudanças significativas nas brânquias. Portanto, juvenis de C. parallelus possuem a capacidade de aclimatação nas salinidades de 5 a 35 semefeitos significativos no desenvolvimento após 60 dias.

Physiological effects of gasoline on the freshwater fish prochilodus lineatus (characiformes: prochilodontidae)

The purpose of this work was to evaluate the effects of the water-soluble fraction of gasoline (WSFG) on the Neotropical freshwater fish Prochilodus lineatus. The WSFG was prepared by mixing gasoline in water (1:4) and animals were exposed for 6, 24 and 96h to 5% diluted WSFG or only to water. After exposure, blood was collected from the caudal vein and the gills were removed. The following parameters were analyzed: hematological (hemoglobin, hematocrit, number of red blood cells), osmo-ionic (plasma Na+, Cl- and K+ and plasma osmolarity), metabolic (total plasma proteins and glucose), endocrine (cortisol), density and distribution of chloride cells [CC] in the gills (immunohistochemistry), and branchial Na+/K+-ATPase (NKA) activity. Hemolysis was found to occur after 96h exposure to WSFG, as indicated by the decrease in the hematological parameters analyzed, followed by an increase in plasma K+. Secondary stress response was revealed by the occurrence of hyperglycemia in the three periods of exposure, despite the absence of significant increases in the plasma cortisol. The exposure to WSFG also caused an increase in the quantity of CC and in plasma Na+, after 24h, as well as in the enzymatic activity of NKA and plasma osmolarity, after 24h and 96h. These results indicate that fish exposed to the WSFG showed physiological adjusts to maintain their osmotic balance. However, the increase in the quantity of CC in the lamellae may interfere in the gas exchange impairing respiration.

O objetivo deste trabalho foi avaliar os efeitos da fração solúvel da gasolina (FSG) em Prochilodus lineatus. A FSG foi preparada misturando-se a gasolina à água (1:4) e os animais foram expostos por 6, 24 e 96h à FSG diluída 5% ou apenas à água. Após a exposição, o sangue foi coletado pela veia caudal e as brânquias foram retiradas. Os seguintes parâmetros foram analisados: hematológicos (hemoglobina, hematócrito, quantidade de células vermelhas), osmo-iônicos (concentrações plasmáticas de Na+, Cl-, K+ e osmolaridade), metabólicos (concentrações plasmáticas de proteínas totais e glicose), endócrino (cortisol plasmático), densidade e distribuição de células-cloreto [CC] nas brânquias (imunohistoquímica) e a atividade da enzima Na+/K+-ATPase (NKA) branquial. Hemólise foi observada após 96h de exposição à FSG, indicada pela diminuição dos parâmetros hematológicos analisados, seguido pelo aumento do K+ plasmático. Houve resposta secundária de estresse visualizada pela ocorrência da hiperglicemia nos três tempos de exposição, apesar da ausência de diferenças significativas na concentração plasmática do cortisol. A exposição à FSG também provocou aumento na quantidade das CC e na concentração de Na+, após 24h, e na atividade enzimática da NKA e osmolaridade, após 24h e 96h. Esses resultados indicam que peixes expostos à FSG apresentaram ajustes fisiológicos para manter o equilíbrio osmótico. Entretanto, o aumento na quantidade de CC nas lamelas pode ter interferido nas trocas gasosas prejudicando a respiração.

IMPROVING THE TOLERANCE OF Vicia faba AGAINST ENVIRONMENTAL SALINITY RESULTED FROM THE IRRIGATION WITH SEA WATER BY USING KNO3 AND (NH4)2SO4 AS CHEMICAL OSMOREGULATORS / Mejoramiento de la tolerancia de Vicia faba a salinidad ocasionada por irrigación con agua de mar usando KNO3 AND (NH4)2SO4 como osmoreguladores químicos

The familiar solutes, Na+, K+, Ca2+, Mg2+, Cl-, PO4(3-), SO4(2-), soluble carbohydrates, amino acids and soluble proteins, which play a role in osmotic adjustment were estimated to investigate the role of potassium nitrate and ammonium sulphate as osmoregulators and their effects on the solutes composition. Vicia faba L. was cultivated and irrigated with 5, 10, 15 and 20 % (v:v) sea water. The plants were divided to three groups. The first was irrigated with sea water only. The second was treated with 5 mM KNO3 while the third was treated with 5mM (NH4)2SO4. The plants were left to grow until flowering stage. The results indicated that the non treated group increased the soluble carbohydrates in the roots to avoid the influx of sodium. The treatment with KNO3 decreased the sodicity (SAR) while (NH4)2SO4 treatment decreased the SK:Na value in the shoots at higher salinity. The availability of nitrogen as nitrate or ammonium ions enhances the accumulation of soluble carbohydrates in shoots. The plants of all groups were depended on Ca2+, as compatible solute more than Na+, and K+.

Los solutos Na+, K+, Ca2+, Mg2+, Cl-, PO4(3-), SO4(2-), carbohidratos y proteínas solubles, así como amino ácidos son importantes en ajuste osmótico y fueron estimados para determinar el papel de nitrato de potasio y sulfato de amonio como osmoreguladores y su efecto en la composición de solutos. Vicia faba L. fue cultivada e irrigada con 5, 10, 15 y 20 % (v:v) de agua de mar permitiendo crecimiento hasta el estado de floración. Las plantas fueron divididas en tres grupos. El primero fue irrigado con agua de mar solamente. El segundo fue tratado con 5 mM KNO3, mientras que el tercer grupo fue expuesto a 5mM (NH4)2SO4. Los resultados indican que las plantas del primer grupo (no tratado) incrementan carbohidratos solubles en sus raíces para evitar influjo de sodio. El tratamiento con KNO3 disminuye riqueza de sodio (SAR) mientras que la exposición a (NH4)2SO4 diminuye la de SK:Na en brotes a salinidad alta. La disponibilidad de nitrato o iones amonio aumenta acumulación en las raíces de carbohidratos solubles. Las plantas de todos los grupos dependen de Ca2+ como soluto compatible más que de Na+ o K+.

Plasma ion levels of freshwater and marine/estuarine teleosts from Southern Brazil

The purpose of this study was to investigate Na+, Cl-, K+, Ca2+, and Mg2+ levels in the plasma of freshwater and marine/estuarine teleosts collected at different salinities (0 to 34) from the estuarine and freshwater portions of the São Gonçalo channel in Southern Brazil. Any relationship between plasma ion levels and salinity and the capacity of ionic regulation of teleosts found at three or more different salinities (Genidens barbus and Micropogonias furnieri) was also investigated. Results showed no relationship between plasma ion levels and salinity when considering all species together, but the two species collected from three or more different salinities showed a significant positive relationship between plasma ion levels and salinity, indicating that G. barbus and M. furnieri have a high capacity to regulate plasma ion levels at both low and high salinities.

A proposta deste estudo foi investigar os níveis de Na+, Cl-, K+, Ca2+, and Mg2+ no plasma de teleósteos de água doce e marinhos/estuarinos coletados em diferentes salinidades (0 a 34) nas porções de água doce e estuarinas do Canal São Gonçalo, sul do Brasil. Uma possível relação entre os níveis iônicos plasmáticos e a salinidade também foi investigada bem como a capacidade de regulação iônica dos teleósteos coletados em três ou mais salinidades (Genidens barbus e Micropogonias furnieri). Os resultados mostraram que não houve relação entre os níveis iônicos no plasma com a salinidade quando considerando todas as espécies juntas, mas as duas espécies coletadas em três ou mais salinidades mostraram uma relação significativamente positiva entre níveis iônicos no plasma e salinidade, mostrando que G. barbus e M. furnieri apresentaram uma grande capacidade para regular os íons do plasma em baixas e altas salinidades.

Dissolved oxygen and ammonia levels in water that affect plasma ionic content and gallbladder bile in silver catfish / Níveis de oxigênio dissolvido e amônia na água afetam o conteúdo iônico do plasma e da bile vesicular em jundiá

Ionic contents (Na+, K+ and Cl-) of plasma and gallbladder bile (GB) of juveniles silver catfish, Rhamdia quelen (156.1±0.2g, 28.2±0.3cm), were determined in three different times (0, 6 and 24h) after exposure to: a) control or high dissolved oxygen (DO = 6.5mg L-1) + low NH3 (0.03mg L-1); b) low DO (3.5mg L-1) + low NH3; c) high DO + high NH3 (0.1mg L-1); and d) low DO + high NH3. High waterborne NH3 or low DO levels increased plasma and GB ion levels. These parameters might have followed different mechanisms to affect osmoregulation since a synergic effect of these variables was detected.

O conteúdo iônico (Na+, K+ e Cl-) do plasma e da bile vesicular (BV) de juvenis de jundiá, Rhamdia quelen (156,1±0,2g, 28,2±0,3cm), foi determinado em três diferentes tempos (0, 6 e 24h) após exposição a: a) controle ou alto oxigênio dissolvido (OD = 6,5mg L-1) + baixa NH3 (0,03mg L-1); b) baixo OD (3,5mg L-1) + baixa NH3; c) alto OD + alta NH3 (0,1mg L-1); e baixo OD + alta NH3 . Alta concentração de amônia ou baixo oxigênio dissolvido na água aumentaram os níveis iônicos no plasma e na BV. Aparentemente, os efeitos osmorregulatórios desses parâmetros podem estar relacionados a mecanismos distintos, pois foi detectado efeito sinérgico sobre essa alteração osmorregulatória.

Metabolic substrates are not mobilized from the osmoregulatory organs (gills and kidney) of the estuarine pufferfishes Sphoeroides greeleyi and S. testudineus upon short-term salinity reduction

The marine-estuarine species of pufferfishes Sphoeroides testudineus and S. greeleyi are very efficient osmoregulators. However, they differ with respect to their tolerance of salinity reduction. During low tide S. testudineus remains in diluted estuarine waters, whereas S. greeleyi returns to seawater (SW). The hypothesis tested here was that the short-term mobilization of metabolic substrates stored in their main osmoregulatory organs would correlate with this differential tolerance. Fishes exposed to 5ë (for 6 h) were compared to those kept in 35ë. Branchial and renal contents of triglycerides, protein and glycogen were evaluated, and total ATPase activity accounted for the tissues' metabolism. Plasma osmolality, chloride and glucose, hematocrit, and muscle water content were also measured. Total triacylglycerol content was higher in S. greeleyi than in S. testudineus in both salinities and in both organs. Kidney glycogen contents were higher in S. greeleyi than in S. testudineus in 5 and 35ë. Total ATPase activity was reduced in 5ë when compared to 35ë in the kidney of S. greeleyi, and was higher in the gills of S. greeleyi than in those of S. testudineus, in both salinities. Upon exposure to dilute SW, both species displayed a similar osmoregulatory pattern: plasma osmolality and chloride were reduced. Again in both species, stability in muscle water content indicated cellular water content control. Although the metabolic substrates stored in the osmoregulatory organs of both species were not mobilized during these short-term sea water dilution events, some differences could be revealed between the two species. S. greeleyi showed more metabolic reserves (essentially triacylglycerols) in these organs, and its gills showed higher total ATPase activity than those S. testudineus.

Os baiacus marinhos Sphoeroides testudineus e S. greeleyi são freqüentadores de estuários e eficientes osmorreguladores. No entanto, eles diferem quanto à sua capacidade de tolerar a redução de salinidade. Durante a descida de maré S. testudineus permanece no estuário com salinidade reduzida e S. greeleyi retorna para a água do mar. A hipótese testada neste estudo foi a de que a mobilização em curto prazo dos substatos metabólicos armazenados nos órgãos osmorregulatórios poderia explicar esta tolerância diferencial a redução de salinidade. Peixes expostos a 5ë (por 6 h) foram comparados com os mantidos em 35ë . O conteúdo branquial e renal de triacilglicerol, proteína e glicogênio foram avaliados, e a atividade ATPásica total foi quantificada para representar o metabolismo do tecido. Osmolalidade, cloreto e glicose plasmáticos, o hematócrito e o conteúdo de água no músculo também foram mensurados. O conteúdo total de triacilglicerol foi maior em S. greeleyi do que em S. testudineus em ambas as salinidades e em ambos os órgãos. O conteúdo renal de glicogênio foi maior em S. greeleyi quando comparado a S. testudineus na duas salinidades. A atividade ATPásica total foi reduzida em 5ë quando comparada a 35ë no rim de S. greeleyi, e foi maior nas brânquias de S. greeleyi quando comparada aos valores para S. testudineus em ambas as salinidades. Após a exposição a água do mar diluída, ambas as espécies apresentaram padrão osmorregulatório similar: osmolalidade e cloreto plasmáticos foram reduzidos. Em ambas as espécies a estabilidade do conteúdo de água no músculo indicou capacidade de regular o conteúdo de água intracelular. Os substratos metabólicos estocados nos órgãos osmorregulatórios de ambas as espécies não foram mobilizados durante a exposição de curto-prazo a diluição da água do mar, porém algumas diferenças foram reveladas entre as duas espécies. S. greeleyi apresentou maior reserva metabólica (essencialmente triacilglicerol) em seus...

Clonamiento y expresión heterologa en saccharomyces cerevisiae de la acuaporina 2 de trypoanosoma cruzy (TcAQP2) / Cloning and heterologous expression in saccharomyces cerevisiae of the aquaporin 2 of trypanosoma cruzi (TcAQ2)

Trypanosoma cruzi es el agente causal de la Enfermedad de Chagas, una entidad endémica no controlada y en crecimiento en Latinoamérica. El parásito, durante su ciclo de vida, se ve sometido a cambios bruscos en la osmolaridad. Tales situaciones de estrés osmótico ameritan sistemas fisiológicos que le permitan a estas células adaptarse y sovrevivir a la nueva situación. En distintos organismos, no de los elementos primordiales que intervienen en osmorregulación son las acuaporinas, proteínas de membrana pertenecientes a la familia MIP ("major intrinsec protein"), que permiten el paso selectivo de agua (acuaparinas clásicas) y otras moléculas no cargadas (acuagliceroporinas) a través de membranas biológicas, en función de gradientes osmóticos. En el genoma de T. cruzi existen cinco genes que presentan similitud con proteínas de la familia MIP. Una de estás ya ha sido caracterizada, TcAQP1 (Trypanosoma cruzi, acuaporina 1), un canal permeable solo al agua. En este artículo de investigación se inició el estudio de un segundo gen con similitud a acuaporina, al cual hemos denominado TcAQP2. El gen TcAQP2 codifica para una proteína de 276 aminoácios. Por medio de herramientas de bioinformática se realizó la predicción de las estructura de dicha proteína; posee características que permiten ubicarla dentro de la familia MIP. Se realizó el clonamiento y secuenciación de la TcAQP2, detectándose diez mutaciones silentes. Este hecho sugiere que se trata de mutaciones propias de la cepa de T. cruzi CL Brener utilizada en el desarrollo experimental de este trabajo. Posteriormente se realizó el subclonamiento de TcAQP2 en el plásmido de expresión en el modelo de levadura pYES.2 y se transformó en las células de saccharomyces cerevisiae. La expresión funcional de la TcAQP2 en dichas células, muestra evidencias de que está proteína interviene en osmorregulación cuando las células son expuestas a choques hiper-osmóticos e hipo-osmóticos...

Trypanosoma cruzi, the etiological agent of changes disease, es prevalent throughout Latin America. During its life cycle, the parasite undergoes extreme fluctuations in osmolarity, consequentrly physiological systems are required to assure its survival. In many organisms one of the more important systems involved in osmoregulation are aquaporins. These proteins are members of the major intrinsic protein family (MIP). Aquaporins can be divided in two groups according to its permeablity: the first one is only permeable to water (orthodox aquiaporins and the second one is permeable to water, glycerol, and other small, uncharged molecules (aquaglyceroporins). In the T cruzi genome there are 5 genes that encode proteins similar to aquaporins. One of these genes (Trypanosoma cruzi aquaporin 1, TcAQp1) has been already characerized as a channel that is only permeable to water. It was also demonstrated that TcAQP1 is involved in parasite osmoregulation. In this work, we started to investigate other gene with aquaporin similarity, which we named Trypanosoma cruzi aquaporin 2 (TcAQP2). The gene TcAQP2 encodes a protein of 276 aminoacids. The protein has six putative transmembrane domains and the two signature motifs asparagine-proline-alanine (NPA), which is the classical conserve motif in the MIP family proteins. Cloning and sequencing of the TcAQP2 gene allowed the detection of ten silent mutations in all clones analyzed, as compare to the sequence reported in data bank of the parasite, suggesting that they belong to the CL Brener strain used in this study. In order to analyze the TcAQP2 function, subcloning in the plasmid for heterologous expression in yeast (pYES.2) of this gene was performed. After transformation and functional expression alf TcAQP2 in Saccharomyces cerevisiae, cells were exposed to osmotic stress. The results indicate that this protein is involved in osmoregulation, and suggest that TcAQP2 participate as a channer for water and/or glycerol in...

The significance of cortisol on acclimation to salinity in pejerrey Odontesthes bonariensis

The role of cortisol on the osmoregulation of pejerrey Odontesthes bonariensis at different salinities was investigated in adult fish injected with 0.7mg hydrocortisone per 100g body weight of fish, and transferred to 0, 5 and 20ppt of NaCl. Blood cortisol was 566ng/ml at the beginning of the experiment (0h) but surged to 1250ng/ml within 3h in cortisol-injected fish. Cortisol levels were influenced not only by treatment but also by time, being higher at 3h compared to 24h. Salinity level, time of exposure and their interaction, but not cortisol treatment, significantly affected plasma osmolality and the concentration of ions Cl- and Na+. This study showed that exogenous cortisol does not seem to play a significant role on the regulation of plasma osmolality and concentration of individual ions in pejerrey

Investigou-se a participação do cortisol na osmoregulação de peixe-rei Odontesthes bonariensis, em diferentes salinidades, em peixes adultos injetados com 0,7mg hidrocortisona por 100g de peso corporal, e transferidos para 0, 5 e 20ppt de NaCl. No inicio do experimento (0h), o cortisol encontrado no plasma foi de 566ng/ml, aumentando para 1250ng/ml em 3h em peixes injetados com cortisol. A concentração de cortisol foi influenciada não somente pelo tratamento, mas também pelo tempo, sendo maior 3h após a inoculação, comparada à 24h. A salinidade, o tempo de exposição e a interação desses dois fatores, mas não o tratamento com o cortisol, afetaram significativamente a osmolaridade e a concentração dos íons Cl- e Na+ do plasma. Este estudo mostrou que, o cortisol exógeno não influi significativamente na regulação da osmolaridade e da concentração de íons Na+ e Cl- no plasma em peixe-rei

Osmoregulation of a Halophilic Bacteria Strain Halomonas sp.BYS-1 / 微生物学通报

Halomonas sp.BYS-1 was a moderately halophilic bacteria strain isolated from activated sludge,It could grow on MM with NaCl concentration from 0.1～2.6 mol/L and phenyl acetic acid as sole carbon souce. When BYS-1 grew in the media with different concentrations of NaCl, there was no obvious change in its intracellular Na+ contents , it accumulated K+, glutamic acid and betaine as osmoprotectants. Its intracellular contents of K+,glutamic acid and betaine increased by 1.9,2.4 and 13.6 times, respectively, when the concentration of NaCl increased from 0.1 mol/L to 2mol/L.