Identification of RSVP14 and RSVP20 components by two-dimensional electrophoresis and Western-blotting.

We have already shown that RSVP14 and RSVP20, two ram seminal plasma (SP) proteins postulated to be involved in sperm capacitation and gamete interaction can protect spermatozoa against cold-shock. In this study, we use two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) for the analysis of SP proteins of Rasa Aragonesa rams, using enhanced protein solubilization in the presence of tributyl phosphine (TBP) and a polyacrylamide linear gradient gel with a narrow pH range (4-7). The image analysis of the 2D map detected 195 protein spots, with isoelectric points (pIs) ranging from 4.5 to 6.6, and molecular weight (M(r)) from 11.7 to 90.4. Staining of 2D gels with Pro-Q Emerald 300 Glycoprotein Stain revealed that most significant proteins in ram SP are glycosylated. The removing of protein N-linked oligosaccharides improved the gel resolution. 2D-PAGE analysis of the whole fraction 6 (F6) separated from ram SP by exclusion chromatography showed six main protein spots, four (a, b, c, d) in the 14 kDa and two (e, f) in the 20 kDa region. Western-blot analyses indicated that the anti-P14 antibody recognized four spots on the SP map, 4, 5, 6 and 7, that matched with spots a, b, c, d of F6 map. The anti-P20 antibody recognized spots 13 and 14 of SP map that corresponded to spots e, f of F6 map. The deduced sequences by de novo sequencing evidenced that protein spots 7 and 13 have significant similarities to BSP family, while protein spots 4 and 14 did not appear to be homologous with any reported protein in the current mammalian Proteinbank databases.     

Effect of Chemical or Electrical Activation of Bovine Oocytes on Blastocyst Development and Quality

Activation of in vitro‐matured (IVM) oocytes is essential for successful embryo production following nuclear transfer (NT) or intracytoplasmic sperm injection (ICSI). This study was designed to compare the rates of blastocyst production and embryo quality (as measured by numbers of viable cells) following parthenogenetic activation with electrical pulse or the use of two different calcium ionophores, A23187 (CA) or ionomycin (IO), with or without the addition of bovine serum albumin (BSA). IVM oocytes with a first polar body were randomly allocated to five treatment groups: CA (5 μm CA, 5 min; n = 88), CA + BSA (5 μm CA, 5 min; BSA, 5 min; n = 90), IO (5 μm IO, 5 min; n = 91), IO + BSA (5 μm IO, 5 min; BSA, 5 min; n = 86) and EL (two pulses of 1.5 kV/cm, 20 μs; n = 120). Blastocyst rates were higher (p < 0.05) for CA (54.4%), IO (51.4%) and EL (54.5%) than for IO + BSA (18.3%). Treatment CA + BSA (39.8%) did not differ from the others. There was no difference (p > 0.05) among treatments in total number of cells. However, the percentage of viable cells was reduced in CA (49.9%), CA + BSA (45.8%), IO (64.9%), IO + BSA (50.9%) compared with EL (82.7%). In summary, the addition of BSA to the IO treatment had an adverse effect on blastocyst production rates. Although there was no difference between electrical stimulation and chemical activation on blastocyst production rates, electrical activation resulted in blastocysts with a higher percentage of viable cells.     

A monolithic photovoltaic-photoelectrochemical device for hydrogen production via water splitting

Direct water electrolysis was achieved with a novel, integrated, monolithic photoelectrochemical-photovoltaic design. This photoelectrochemical cell, which is voltage biased with an integrated photovoltaic device, splits water directly upon illumination; light is the only energy input. The hydrogen production efficiency of this system, based on the short-circuit current and the lower heating value of hydrogen, is 12.4 percent.     

Cross-protective immunity and the serological classification system for Bacteroides nodosus

The relationship between the serological classification system for serogroup B and for serogroup H of Bacteroides nodosus and cross-protection between subgroups within these serogroups was examined. Protection against ovine footrot following vaccination was achieved against other subgroup strains provided sufficient cross-reactive antibody was induced by shared pilus antigens. Within serogroup B, better cross-protection against one subgroup was obtained with a pili vaccine than a whole cell vaccine which correlated with higher pilus antibody titres induced by the former. For serogroup H, a lack of cross-protection and serological reactivity between subgroups was demonstrated, which indicates that the prototype strain of subgroup H2 should be designated a new serogroup.     

Relationship of blood corticosterone, immunoglobulin and haematological values in young crocodiles (Crocodylus porosus) to water temperature, clutch of origin and body weight

Objective To examine whether sub-optimal temperature induced stress and immunosuppression in farmed saltwater crocodile (Crocodylus porosus) hatchlings. Design A clinico-pathological study. Animals A total of 140 hatchlings were used. Procedure Body weight and length, plasma corticosterone and immunoglobulin concentrations and total and differential white blood cell counts were measured in 140 hatchlings from five clutches divided between five water temperature treatment groups. Initially all groups were housed at 32°C for 10 weeks, then two groups (L, LC) were changed to low temperature (28°C) and two groups (H, HC) to high temperature (36°C), while one group (C) remained at 32°C. The LC and HC groups were maintained at these temperatures for 10 days, after which the water temperature of both groups was returned to 32°C. Blood samples were collected twice (at 6 and 9 weeks of age) before the initial temperature change, and at 10 days and 4 weeks after the initial temperature change (at 11.5 and 14 weeks of age). Results Except for an increase in plasma corticosterone in the HC group and a decrease in the L group when the temperature change was first introduced, changes in plasma corticosterone were not significant. There were no significant changes in immunoglobulin concentrations. There were, however, significant decreases in the total white cell and lymphocyte counts in the LC group after the temperature was decreased to 28°C, and an increase in these counts after water temperature was returned to 32°C. Clutch of origin had significant effects on body weight and length gains, and there were negative relationships between body weight and corticosterone concentrations and between body weight and immunoglobulin concentrations. Conclusions As haematological changes indicative of stress were not associated with significant changes in serum corticosterone, immunosuppression in young crocodiles may be independent of the hypothalamic-pituitary-adrenal cortical axis.