NEW RECORD OF CALYPTRAEID HOSTS FOR THE PEA CRAB CALYPTRAEOTHERES GARTHI (FENUCCI, 1975) (DECAPODA, PINNOTHERIDAE) IN ARGENTINA

Calyptraeotheres garthi (Fenucci, 1975) is one of nine species of Pinnotheridae occurring in Argentina and the only species of that genus located in this region. This species can be found from the San Maths Gulf, Argentina to Rio Grande do Sul, Brazil where it lives in symbiotic association with limpets of the family Calyptraeidae. Currently, two species of limpets have been reported as hosts of this species: Crepidula protea (d'Orbigny, 1841) and Crepidula argentina Simone, Pastorino & Penchaszadeh, 2000. We report here four more species of limpets serving as hosts of C. garthi, namely Bostrycapulus odites Collin, 2005, Crepidula cachimilla Cledon, Simone & Penchaszadeh, 2004, Crepidula plana Say, 1822 and Trochita pileus (Lamarck, 1822). Thus, this pinnotherid appears to be, as others members of the family, a generalist symbiotic species since it dwells on almost all the potential limpet species reported for Argentina. However, there are notable differences in prevalence of C. garthi on each host, which suggest that the symbiont might prefer species of Crepidula over the others genera (Trochita and Bostrycapulus). Additionally, there exists an apparent relationship between host size and prevalence (e.g., Crepidula cachimilla, the limpet with the highest prevalence, is clearly larger than the others hosts). Perhaps it is premature to conclude that the size of the host might be of benefit for the symbiont, but it is a hypothesis worthy of being explored.

Effects of bovine sperm cryopreservation using different freezing techniques and cryoprotective agents on plasma, acrosomal and mitochondrial membranes

The success of semen cryopreservation is influenced by several factors, such as freezing curves and cryoprotectants. These two factors are of special interest once they may lead to many important physical-chemical changes resulting in different degrees of damage in spermatozoa structure. This experiment was designed to compare the effect of bull semen cryopreservation using two freezing techniques: conventional (CT cooling rate of -0.55 degrees C min-1 and freezing rate of -19.1 degrees C min-1) and automated (AT cooling rate of -0.23 degrees C min-1 and freezing rate of -15 degrees C min-1), performed with different curves, and with three cryoprotectants (glycerol, ethylene glycol and dimethyl formamide) on bovine sperm motility and integrity of plasma, acrosomal and mitochondrial membranes. These variables were simultaneously evaluated using the fluorescence probes propidium iodide, fluorescein-conjugated Pisum sativum agglutinin and MitoTracker Green FM. The effects of freezing techniques, as well as of different cryoprotectants were analysed by the analysis of variance. The means were compared by Fishers test. There were no significant differences between freezing techniques (P > 0.05). Glycerol showed higher percentages of motility, vigour and integrity of plasma, acrosomal and mitochondrial membranes than other two cryoprotectants (P < 0.05). Ethylene glycol preserved higher motility and integrity of plasma and mitochondrial membranes than dimethyl formamide (P < 0.05). Sperm motility with glycerol was 30.67 +/- 1.41% and 30.50 +/- 1.06%, with ethylene glycol was 21.17 +/- 1.66% and 21.67 +/- 1.13% and with dimethyl formamide was 8.33 +/- 0.65% and 9.17 +/- 0.72% to CT and AT curves, respectively. The percentage of spermatozoa with simultaneously intact plasma membrane, intact acrosome and mitochondrial function (IPIAH) was 14.82 +/- 1.49% (CT) and 15.83 +/- 1.26% (AT) to glycerol, 9.20 +/- 1.31% (CT) and 9.92 +/- 1.29% (AT) to ethylene glycol 4.65 +/- 0.93% (CT) and 5.17 +/- 0.87% (AT) to dimethyl formamide. Glycerol provided the best results, although nearly 85% of spermatozoa showed some degree of injury in their membranes, suggesting that further studies are required to improve the results of cryopreservation of bovine semen.

Effects of discontinuous Percoll gradient centrifugation on the quality of bovine spermatozoa evaluated with computer-assisted semen analysis and fluorescent probes association

The objective of this study was to evaluate the quality of bovine frozen-thawed sperm cells after Percoll gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Four ejaculates per bull were evaluated before and after discontinuous Percoll gradient centrifugation. Sperm motility was assessed by computer-assisted semen analysis and the integrity of the plasma and acrosomal membranes, as well as mitochondrial function, were evaluated using a combination of fluorescent probes propidium iodide, fluorescein isothiocyanate-conjugated Pisum sativum agglutinin and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide. The procedure of Percoll gradient centrifugation increased the percentage of total and progressive sperm motility, beat frequency, rectilinear motility, linearity and rapidly moving cells. In addition, the percentage of cells with intact plasma membrane and mitochondrial membrane potential was increased in post-centrifugation samples. However, the percentage of sperm cells with intact acrosomal membrane was markedly reduced. The method used selected the motile cells with intact plasma membrane and higher mitochondrial functionality in frozen-thawed bull semen, but processing, centrifugation and/or the Percoll medium caused damage to the acrosomal membrane.

Biochemical, physicochemical and molecular characterization of a genuine 2-Cys-peroxiredoxin purified from cowpea [Vigna unguiculata (L.) Walpers] leaves

Background: Peroxiredoxins have diverse functions in cellular defense-signaling pathways. 2-Cys-peroxiredoxins (2-Cys-Prx) reduce H2O2 and alkyl-hydroperoxide. This study describes the purification and characterization of a genuine 2-Cys-Prx from Vigna unguiculata (Vu-2-Cys-Prx). Methods: Vu-2-Cys-Prx was purified from leaves by ammonium sulfate fractionation, chitin affinity and ion exchange chromatography. Results: Vu-2-Cys-Prx reduces H2O2 using NADPH and DTT. Vu-2-Cys-Prx is a 44 kDa (SDS-PAGE)/46 kDa (exclusion chromatography) protein that appears as a 22 kDa molecule under reducing conditions, indicating that it is a homodimer linked intermolecularly by disulfide bonds and has a pI range of 4.56-4.72; its NH2-terminal sequence was similar to 2-Cys-Prx from Phaseolus vulgaris (96%) and Populus tricocarpa (96%). Analysis by ESI-Q-TOF MS/MS showed a molecular mass/pI of 28.622 kDa/5.18. Vu-2-Cys-Prx has 8% alpha-helix, 39% beta-sheet, 22% of turns and 31% of unordered forms. Vu-2-Cys-Prx was heat stable, has optimal activity at pH 7.0, and prevented plasmid DNA degradation. Atomic force microscopy shows that Vu-2-Cys-Prx oligomerized in decamers which might be associated with its molecular chaperone activity that prevented denaturation of insulin and citrate synthase. Its cDNA analysis showed that the redox-active Cys(52) residue and the amino acids Pro(45), Thr(49) and Arg(128) are conserved as in other 2-Cys-Prx. General significance: The biochemical and molecular features of Vu-2-Cys-Prx are similar to other members of 2-Cys-Prx family. To date, only one publication reported on the purification of native 2-Cys-Prx from leaves and the subsequent analysis by N-terminal Edman sequencing, which is crucial for construction of stromal recombinant 2-Cys-Prx proteins. (C) 2012 Elsevier B.V. All rights reserved.

Potencial fisiológico de sementes armazenadas e desempenho de plantas de ervilha

Os efeitos do vigor de lotes de sementes sobre a emergência das plântulas e estabelecimento do estande, principalmente sob condições menos favoráveis de ambiente, estão bem documentados na literatura. Porém, há necessidade de intensificar a pesquisa para esclarecer as relações entre o potencial fisiológico das sementes e o desempenho das plantas em campo; este foi o principal objetivo deste estudo.Utilizaram-se duas cultivares de ervilha, 'Telefone Alta' (crescimento indeterminado) e 'Itapuã' (determinado), cada uma representada por quatro lotes armazenados, durante oito meses, em três ambientes: laboratório, câmara fria e seca (10 ºC e 30% de umidade relativa do ar) e ambiente controlado (20 ºC e 70% de umidade relativa do ar); este procedimento permitiu criar diferenças entre o potencial fisiológico dos lotes de cada cultivar. Após a determinação da germinação e do vigor (condutividade elétrica, envelhecimento acelerado, comprimento e emergência de plântulas), foi conduzido ensaio de campo, realizando-se avaliações do estande inicial e final, altura e área foliar de plantas, número de vagens e produção de grãos verdes e secos. O vigor das sementes de ervilha influencia a emergência de plântulas e o estabelecimento do estande em campo, especialmente em lotes pouco vigorosos. O vigor de sementes de ervilha afeta negativamente o desenvolvimento das plantas e a produção final, quando há redução acentuada do estande; a extensão desses efeitos é proporcional à intensidade dessa redução.