Does Nest Predation Influence Colony Movements of Saunders's Gulls (Saundersilarus saundersi) in a Reclaimed Land Area?

Some seabirds commonly use artificially reclaimed lands, which are frequently located next to mainland environments, for breeding. Nest predation risk caused by birds or mammals from the mainland has negative influence on fitness-related costs and distribution of seabirds. Here, we sought to link potential factors, specifically those related to nest predation and nest environment, with breeding performance and colony movements of the Saunders's gull (Saundersilarus saundersi), a vulnerable species, on a large reclaimed area (1350 ha) in Incheon in Republic of Korea. This reclaimed area has experienced rapid changes in communities of nest predators from the mainland and vegetation ranging from halophytes to terrestrial plants after reclamation. Additionally, changes in the surrounding of used nest sites were retrospectively examined to determine whether colony movement was reversible in this reclaimed area. Our results indicated that high nest predation in a previous year induced colony movements in a consecutive year while the breeding colony exhibited a gradual reduction in clutch size. However, such movement after high nest predation seemed to be irreversible due to ongoing habitat degradation caused by construction and vegetation alteration. This study highlights that high nest predation may exert strong pressure on breeding colonies of Saunders's gulls. It also has anthropogenic impacts, leading to continuous dispersal of colonies to new areas for this vulnerable seabird in a reclaimed land.

Nest predation risk influences a cavity-nesting passerine during the post-hatching care period.

Some nest predators visually assess parental activities to locate a prey nest, whereas parents modify fitness-related traits to reduce the probability of nest predation, and/or nestlings fledge early to escape the risky nest environment. Here, we experimentally tested if the parental and fledging behaviours of oriental tits (Parus minor) that bred in the nest-box varied with cavity conditions associated with nest predation risk during the nestling period. The entrance of experimental nest-boxes was enlarged to create a long-term risk soon after clutch competition. A short-term risk, using simulated playbacks with a coexisting control bird and avian nest predator sound, was simultaneously applied to the nest-boxes whether or not the long-term risk existed. We found that the parents reduced their hourly feeding trips, and the nestlings fledged early with the long-term risk, although the nest mortality of the two nest-box types was low and did not differ. While this study presents a portion of prey-predator interactions with the associated uncertainties, our results highlight that the entrance size of cavities for small hole-nesting birds may play an important role in determining their fitness-related traits depending upon the degree of perceived risk of nest predation.

Sexual differences in post-hatching Saunders's gulls: size, locomotor activity, and foraging skill.

Various selection pressures induce the degree and direction of sexual size dimorphism in animals. Selection favors either larger males for contests over mates or resources, or smaller males are favored for maneuverability; whereas larger females are favored for higher fecundity, or smaller females for earlier maturation for reproduction. In the genus of Larus (seagulls), adult males are generally known to be larger in size than adult females. However, the ontogeny of sexual size dimorphism is not well understood, compared to that in adults. The present study investigates the ontogeny of sexual size dimorphism in Saunders's gulls (Larus saundersi) in captivity. We artificially incubated fresh eggs collected in Incheon, South Korea, and measured body size, locomotor activity, and foraging skill in post-hatching chicks in captivity. Our results indicated that the sexual differences in size and locomotor activity occurred with the post-hatching development. Also, larger males exhibited greater foraging skills for food acquisition than smaller females at 200 days of age. Future studies should assess how the adaptive significance of the sexual size dimorphism in juveniles is linked with sexual divergence in survival rates, intrasexual contests, or parental effort in sexes.

A proteomic approach for identifying cellular proteins interacting with erythropoietin in recombinant Chinese hamster ovary cells.

Identification of the cellular proteins interacting with incompletely folded and unfolded forms of erythropoietin (EPO) in recombinant CHO (rCHO) cells leads to better insight into the possible genetic manipulation approaches for increasing EPO production. To do so, a pull-down assay was performed with dual-tagged (N-terminal GST- and C-terminal hexahistidine-tagged) EPO expressed in E. coli as bait proteins and cell lysates of rCHO cells (DG44) as prey proteins. Cellular proteins interacting with dual-tagged EPO were then resolved by two-dimensional gel electrophoresis (2DE) and identified by MALDI-TOF MS/MS. A total of 27 protein spots including glucose-regulated protein 78 (GRP78) were successfully identified. Western blot analysis of GRP78 confirmed the results of the MS analyses. Taken together, a pull-down assay followed by a proteomic approach is found to be an efficient means to identify cellular proteins interacting with foreign protein in rCHO cells.

Limitations to the comparative proteomic analysis of thrombopoietin producing Chinese hamster ovary cells treated with sodium butyrate.

Sodium butyrate (NaBu) is known to enhance the specific productivity of Chinese hamster ovary cells expressing human thrombopoietin. In order to better understand the intracellular responses of these cells resulting from NaBu treatment, the proteomic profiles of cells treated with various concentrations of NaBu (0-3mM) were compared using two-dimensional electrophoresis (2-DE). Based on spot intensities, 80 high intensity protein spots were selected. Fifty-six of the 80 protein spots, which represent 28 different kinds of proteins, were identified by MALDI-TOF-MS and MS/MS. Compared to control without NaBu treatment, the expression levels of 2 proteins (glucose regulated protein 78 (GRP 78) and peroxiredoxin 4) were increased over two fold with NaBu treatment and the expression level of phosphopyruvate hydratase was decreased over two fold with NaBu treatment. Due to multiplicity (multiple spots for one protein), a change in one single spot intensity from a 2-DE gel image may not represent the total change in expression level for that protein. Western blot analyses of GRP78, HSC70 and ERp57 confirmed the results of the MS analyses. However, a degree of change in expression level differed between the two methods, suggesting the necessity of a validating method to determine the total amount of the protein.

Proteome analysis of cellular response of Pseudomonas putida KT2440 to tetracycline stress.

Tetracycline-induced proteome of Pseudomonas putida KT2440 was analyzed by 2-D gel electrophoresis and matrix-assisted laser desorption ionization-time of flight/mass spectrum (NALDI-TOF/MS) in order to understand cellular response to tetracycline. Of the proteins upregulated in a culture medium containing subinhibitory concentration of tetracycline (50 mug/mL), we identified 38 proteins from cytosol and precipitated fractions by peptide mass fingerprinting and mass spectrum/mass spectrum analysis. Various amino acids ABC transporters, a ribose ABC transporter, and a sulfate ABC transporter were found to be upregulated. Protein synthesis-related proteins, stress proteins, energy metabolic enzymes, and unknown proteins were also strongly induced. Of the identified upregulated proteins, several proteins (isocitrate lyase, branched-chain amino acid ABC transporter, superoxide dismutase, etc.) were also upregulated under phenol-induced stress condition. These results demonstrate that tetracycline at a high concentration induced comprehensive stress in P. putida KT2440 and the global induction of proteins related to bacteria survival. Proteome analysis was found to be a useful tool for the elucidation of antibiotic-induced proteins in the present study.

Initial transcriptome and proteome analyses of low culture temperature-induced expression in CHO cells producing erythropoietin.

Low culture temperature is known to enhance the specific productivity of Chinese hamster ovary (CHO) cells expressing erythropoietin (EPO) (LGE10-9-27). Genomic and proteomic approaches were taken to better understand the intracellular responses of these CHO cells resulting from use of low culture temperature (33 degrees C). For transcriptome analysis, commercially available rat and mouse cDNA microarrays were used. The data obtained from the rat and mouse cDNA chips were only somewhat informative in understanding the gene expression profile of CHO cells because of their different sequence homologies with CHO transcriptomes. Overall, transcriptome analysis revealed that low culture temperature could lead to changes in gene expression in various cellular processes such as metabolism, transport, and signaling pathways. Proteome analysis was carried out using 2-D PAGE. Based on spot intensity, 60 high intensity protein spots, from a total of more than 800, were chosen for MS analysis. Forty of the 60 protein spots, which represent 26 different kinds of proteins, were identified by MALDI-TOF-MS and validated by MS/MS. Compared to the reference temperature (37 degrees C), the expression levels of seven proteins (PDI, vimentin, NDK B, ERp57, RIKEN cDNA, phosphoglycerate kinase, and heat shock cognate 71 kDa protein) were increased over twofold at 33 degrees C and those of two proteins (HSP90-beta and EF2) were decreased over twofold at 33 degrees C. Taken together, the results demonstrate the potential of combined analysis of transcriptome and proteome analyses as a tool for the systematic comprehension of cellular mechanisms in CHO cells.