Preliminary studies on the vitrification of red sea bream (Pagrus major) embryos

The objective was to identify an appropriate cryoprotectant and protocol for vitrification of red sea bream (Pagrus major) embryos. The toxicity of five single-agent cryoprotectants, dimethyl sulfoxide (DMSO), propylene glycol (PG), ethylene glycol (EG), glycerol (GLY), and methyl alcohol (MeOH), as well as nine cryoprotectant mixtures, were investigated by comparing post-thaw hatching rates. Two vitrifying protocols, a straw method and a solid surface vitrification method (copper floating over liquid nitrogen), were evaluated on the basis of post-thaw embryo morphology. Exposure to single-agent cryoprotectants (10% concentration for 15 min) was not toxic to embryos, whereas for higher concentrations (20 and 30%) and a longer duration of exposure (30 min), DMSO and PG were better tolerated than the other cryoprotectants. Among nine cryoprotectant mixtures, the combination of 20% DMSO + 10% PG + 10% MeOH had the lowest toxicity after exposure for 10 min or 15 min. High percentages of morphologically intact embryos, 50.6 +/- 16.7% (mean +/- S.D.) and 77.8 +/- 15.5%, were achieved by the straw vitrifying method (20.5% DMSO + 15.5% acetamide + 10% PG, thawing at 43 degrees C and washing in 0.5 M sucrose solution for 5 min) and by the solid surface vitrification method (40% GLY, thawing at 22 degrees C and washing in 0.5 M sucrose solution for 5 min). After thawing, morphological changes in the degenerated embryos included shrunken yolks and ruptured chorions. Furthermore, thawed embryos that were morphologically intact did not consistently survive incubation. (C) 2007 Elsevier Inc. All rights reserved.

Corrosion of steel structures in sea-bed sediment

Seabed sediment (SBS) is a special soil that is covered by seawater. With the developments in marine oil exploitation and engineering, more and more steel structures have been buried in SBS. SBS corrosion has now become a serious problem in marine environment and an important issue in corrosion science. In this paper, approach in the field of SBS corrosion is reviewed. Electrochemical and microbial corrosion factors, corrosion mechanism, measurement of metal corrosion rate, corrosion evaluation and prediction of corrosion are also discussed here.

Corrosion failure of marine steel in sea-mud containing sulfate reducing bacteria

The corrosion failure behavior of marine steel is affected by stress, which exists in offshore structures at sea-mud region. The sulfate reducing bacteria (SRB) in the sea-mud made the steel more sensitive to stress corrosion cracking (SCC) and weaken the corrosion fatigue endurance. In this paper, a kind of natural sea-mud containing SRB was collected. Both SCC tests by slow strain rate technique and corrosion fatigue tests were performed on a kind of selected steel in sea-mud with and without SRB at corrosion and cathodic potentials. After this, the electrochemical response of static and cyclic stress of the specimen with and without cracks in sea-mud was analyzed in order to explain the failure mechanism. Hydrogen permeation tests were also performed in the sea-mud at corrosion and cathodic potentials. It is concluded that the effect of SRB on environment sensitive fracture maybe explained as the consequences of the acceleration of SRB on corrosion rate and hydrogen entry into the metal.

Effects of anaerobe in sea bottom sediment on the corrosion of carbon steel

The in-situ study of steel corrosion in sea bottom sediment (SBS) was carried out by Transplanting Burying Plate method (TBP method). It was found that the corrosion rate of steel in the sea bottom sediment with sulfate reducing bacteria (SRB) could be as high as ten times of that in sea bottom sediment without SRB. The experiments in simulated sea bottom sediments with different SRB contents by artificial culturing showed that the electrochemical behavior of steel in the sea bottom sediment with SRB was different from that without SRB. SRB altered the polarization behavior of steel significantly. The environment was acidified due to the activity of SRB and the corrosion of steel was accelerated. The corrosion of carbon steel in sea bottom sediment is anaerobic corrosion, and the main factor is anaerobe. There are SRB commonly in SBS, and the amount of SRB decreases along with the depth of sediment. Because of the asymmetry and variation of sea bottom sediment, the most dangerous corrosion breakage of steel in SBS is local corrosion caused by SRB. So the main countermeasure of corrosion protection of sea bottom steel facilities should be controlling of the corrosion caused by anaerobe.

Alloy elements' effect on anti-corrosion performance of low alloy steels in different sea zones

The corrosion rate of low alloy steel in different sea zones has close correlation with the content of the alloy element. From the field data of steel corrosion rates in atmospheric zone, splash zone and immersion zone, regression analysis was used to study the correlation between the corrosion rate of steels and the amount of added alloy elements. Three regression equations were obtained in different sea zones. Based on the equations, the anti-corrosion performance of the alloy elements can be deduced which can be used to screen out low alloy steel with good anti-corrosion performance. (C) 2007 Elsevier B.V. All rights reserved.

Effect of sulfate-reducing bacteria on corrosion behavior of mild steel in sea mud

Microbiologically influenced corrosion (MIC) is very severe corrosion for constructions buried under sea mud environment. Therefore it is of great importance to carry out the investigation of the corrosion behavior of marine steel in sea mud. In this paper, the effect of sulfate-reducing bacteria (SRB) on corrosion behavior of mild steel in sea mud was studied by weight loss, dual-compartment cell, electronic probe microanalysis (EPMA), transmission electron microscopy (TEM).combined with energy dispersive X-ray analysis (EDX) and electrochemical impedance spectroscopy (EIS). The results showed that corrosion rate and galvanic current were influenced by the metabolic activity of SRB. In the environment of sea mud containing SRB, the original corrosion products, ferric (oxyhydr) oxide, transformed to iron sulfide. With the excess of the dissolved H2S, the composition of the protective layer formed of FeS transformed to FeS2 or other non-stoichiometric polysulphide, which changed the state of the former layer and accelerated the corrosion process.

The hydrogen permeation investigation of API X56 steel in sea mud

It was found that the corrosion rate of steel in the sea mud with sulfate-reducing bacteria (SRB) could be as high as 10 times of that in the sea mud without SRB. And the hydrogen permeation reaction would occur when metals were corroded. So it is necessary to investigate the effect of living SRB on hydrogen permeation in the sea mud. Cathodic potential was often added to metals in order to protect them. But hydrogen permeation could be affected by the cathodic potential. So it is also necessary to study the effect of cathodic potential on hydrogen permeation. In this paper, the hydrogen permeation actions of APT X56 steel in the sea mud with and without SRB at corrosion and cathodic potential were studied with an improved Devanathan-Stachurski's electrolytic cell. Experimental results showed that during the growth of SRB, the current density curve of hydrogen permeation was accordant with the growth curve of SRB. But the hydrogen permeation current density of APT X56 steel hardly changed in the sterilized sea mud. Compared with the hydrogen permeation current density of APT X56 steel in the sterilized sea mud, the hydrogen permeation of APT X56 steel in the sea mud could be accelerated by living SRB. Experimental results also showed that the hydrogen permeation current density increased rapidly when the cathodic potential was added to the three-electrode system of the cathodic cell, and then the hydrogen permeation current density could obtain a stable value slowly. So the cathodic potential added to the cathodic cell could accelerate hydrogen permeation.

Diversities in hepatic HIF-1, IGF-I/IGFBP-1, LDH/ICD, and their mRNA expressions induced by CoCl2 in Qinghai-Tibetan plateau mammals and sea level mice

Ochotona curzoniae and Microtus oeconomus are the native mammals living on the Qinghai-TibetanPlateau of China. The molecular mechanisms of their acclimatization to the Plateau-hypoxia remain unclear. Expressions of hepatic hypoxia-inducible factor (HIF)-1 alpha, insulin-like growth factor-I (IGF-I)/IGF binding protein (BP)-1(IGFBP-1; including genes), and key metabolic enzymatic genes [lactate dehydrogenase (LDH)-A/isocitrate dehydrogenase (ICD)] are compared in Qinghai-Tibetan- Plateau mammals andsea- level mice after injection of CoCl2 (20, 40, or 60 mg/ kg) and normobaric hypoxia (16.0% O-2, 10.8% O-2, and 8.0% O-2) for 6 h, tested by histochemistry, Western blot analysis, ELISA, and RT-PCR. Major results are CoCl2 markedly increased 1) HIF-1 alpha only in mice, 2) hepatic and circulatory IGF-I in M. oeconomus, 3) hepatic IGFBP-1 in mice and O. curzoniae, and 4) LDH-A but reduced ICD mRNA in mice (CoCl2 20 mg/kg) but were unchanged in the Tibetan mammals. Normobaric hypoxia markedly 1) increased HIF-1 alpha and LDH-A mRNA in mice and M. oeconomus (8.0% O-2) not in O. curzoniae, and 2) reduced ICD mRNA in mice and M. oeconomus (8.0% O-2) not in O. curzoniae. Results suggest that 1) HIF-1 alpha responsiveness to hypoxia is distinct in lowland mice and plateau mammals, reflecting a diverse tolerance of the three species to hypoxia; 2) CoCl2 induces diversities in HIF-1, IGF-I/IGFBP-1 protein or genes in mice, M. oeconomus, and O. curzoniae. In contrast, HIF-1 mediates IGFBP-1 transcription only in mice and in M. oeconomus (subjected to severe hypoxia); 3) differences in IGF-I/IGFBP-1 expressions induced by CoCl2 reflect significant diversities in hormone regulation and cell protection from damage; and 4) activation of anaerobic glycolysis and reduction of Krebs cycle represents strategies of lowland-animals vs. the stable metabolic homeostasis of plateau- acclimatized mammals.