Seasonality influence on biochemical and hematological indicators of stress and growth of pirarucu (Arapaima gigas), an Amazonian air-breathing fish.

Environmental factors such as seasonal cycles are the main chronic stress cause in fish increasing incidence of disease and mortality and affecting productive performance. Arapaima gigas (pirarucu) is an Amazonian air-breathing and largest freshwater fish with scales in the world. The captivity development of pirarucu is expanding since it can fatten up over 1 kg per month reaching 10 kg body mass in the first year of fattening. This work was conducted in three periods (April to July 2010, August to November 2010, and December 2010 to March 2011) defined according to rainfall and medium temperatures. Seasonality effect analysis was performed on biochemical (lectin activity, lactate dehydrogenase, and alkaline phosphatase activities) and hematological (total count of red blood cells, hematocrit, hemoglobin, and hematimetric Wintrobe indexes) stress indicators, as well as on growth and wellbeing degree expressed by pirarucu condition factor developed in captivity. All biochemical and hematological stress indicators showed seasonal variations. However, the fish growth was allometrically positive; condition factor high values indicated good state of healthiness in cultivation. These results reinforce the robust feature of pirarucu and represent a starting point for understanding stress physiology and environmental changes during cultivation enabling identification and prevention of fish adverse health conditions.

Immunomodulatory response of mice splenocytes induced by RcaL, a lectin isolated from cobia fish (Rachycentron canadum) serum.

This work reports the isolation of a serum lectin from cobia fish (Rachycentron canadum) named RcaL. Immunomodulatory activity on mice splenocyte experimental cultures through cytotoxic assays and cytokine production were also performed. RcaL was obtained through precipitation with ammonium sulphate and affinity chromatography on a Concanavalin A-Sepharose 4B column. The ammonium sulphate fraction F3 showed the highest specific hemagglutinating activity and was applied to affinity chromatography. The lectin was eluted with methyl-α-D-mannopyranoside. RcaL showed highest affinity for methyl-α-D-mannopyranoside and D-mannose; eluted fractions of RcaL agglutinated rabbit erythrocytes (titre, 128(-1)) retained 66 % of chromatographed lectin activity, and the obtained purification factor was 1.14. Under reducing conditions, a polypeptide band of 19.2 kDa was revealed in sodium dodecyl sulphate polyacrylamide gel electrophoresis (PAGE). PAGE confirmed RcaL as an acidic protein revealed in a single band. Cytotoxic and immunomodulatory assays with RcaL in mice splenocyte cultures showed that the lectin was not cytotoxic and induced higher interferon gamma and nitric oxide production in splenocyte cultures. Purified RcaL induced preferential Th1 response, suggesting that it acts as an immunomodulatory compound.

Purification and characterization of a mannose recognition lectin from Oreochromis niloticus (tilapia fish): cytokine production in mice splenocytes.

The aim of this work was to purify and partially characterize a mannose recognition lectin from Nile tilapia (Oreochromis niloticus) serum, named OniL. OniL was isolated through precipitation with ammonium sulfate and affinity chromatography (Concanavalin A-Sepharose 4B). In addition, we evaluated carbohydrate specificity, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) profiles, and in vitro immunomodulatory activity on mice splenocyte experimental cultures through cytotoxic assays and cytokine production. The ammonium sulfate fraction F2 showed the highest specific hemagglutinating activity (331) and was applied to affinity matrix. Adsorbed proteins (OniL) were eluted with methyl-α-D: -mannopyranoside. OniL, a 17-kDa protein by SDS-PAGE constituted by subunits of 11 and 6.6 kDa, showed highest affinity for methyl-α-D: -mannopyranoside and D: -mannose. Immunological assays, in vitro, showed that OniL did not show cytotoxicity against splenocytes, induced higher IFN-γ production and lower IL-10 as well as nitrite release. In conclusion, OniL lectin was successfully purified and showed a preferential Th1 response in mice splenocytes.

Desenvolvimento ovariano de Plagioscion squamosissimus (Heckel, 1840) (Actinopterygii, Perciformes), no reservatório de Pedra, Rio de Contas, Bahia / Ovarian development of Plagioscion squamosissimus (Heckel, 1840) (Actinopterygii, Perciformes), from the Pedra reservoir, Contas river, Bahia

O presente estudo tem por objetivo caracterizar a organização dos ovócitos e os estádios maturacionais do ovário da pescada Plagioscion squamosissimus. As coletas foram realizadas bimestralmente no reservatório de Pedra, rio de Contas (BA), entre novembro de 2004 e setembro de 2006. As análises microscópicas foram realizadas através de cortes histológicos, corados com hematoxilina - eosina - floxina e mistura tricrômica de Gomori. Através da análise macro e microscópica, foram identificados cinco estádios maturacionais. A coloração das gônadas variou de acordo com o desenvolvimento gonadal. Para o desenvolvimento ovocitário, foram definidas seis fases: ovogônia e perinucleolar; vitelogênica e lipídica inicial; vitelogênica e lipídica intermediária; vitelogênica e lipídica avançada e pré-ovulação. Foram observadas estruturas semelhantes às de peixes marinhos, como fusão de gotículas de óleo nas fases finais de maturação e hidratação pré-ovulatória. Estas características podem ser explicadas pela origem marinha da família Sciaenidae, indicando a necessidade de estudos mais detalhados para compreender o aparecimento, desenvolvimento e função dessas estruturas ao longo do desenvolvimento ovocitário.

This study aimed to characterize oocyte organization and maturation stages of Plagioscion squamosissimus ovary. Samples were collected bimonthly in Pedra reservoir, Contas River (BA), between November 2004 and September 2006. Microscopic analyses were made on histological cuts stained with hematoxylin - eosin - phloxine and Gomori trichromic mixture. Five maturation stages were identified through macro and microscopic analysis. Gonad coloration varied according to maturation stages. Six phases of oocyte development were identified: oogonia and perinucleolar, vitellogenic and early lipidic, vitellogenic and intermediate lipidic, vitellogenic and advanced lipidic, and pre-ovulation. Cell structures were similar to those found among marine species, such as oil droplets fusion at later developmental stages and pre-ovulatory hydration. Such features may be explained by the marine origin of Sciaenidae, thus suggesting the need of more detailed studies for understanding the origin, development and function of these structures along oocyte developmental stages.