First estimates of metabolic rate in Atlantic bluefin tuna larvae

Atlantic bluefin tuna is an iconic scombrid species with a high commercial and ecological value. Despite their importance, many physiological aspects, especially during the larval stages, are still unknown. Metabolic rates are one of the understudied aspects in scombrid larvae, likely due to challenges associated to larval handling before and during respirometry trials. Gaining reliable estimates of metabolic rates is essential to understand how larvae balance their high growth needs and activity and other physiological functions, which can be very useful for fisheries ecology and aquaculture. This is the first study to (a) estimate the relationship between routine metabolic rate (RMR) and larval dry weight (DW) (mass scaling exponent) at a constant temperature of 26°C, (b) measure the RMR under light and darkness and (c) test whether the interindividual differences in the RMR are related to larval nutritional status (RNA/DNA and DNA/DW). The RMR scaled nearly isometrically with body size (b = 0.99, 0.60–31.56 mg DW) in contrast to the allometric relationship observed in most fish larvae (average b = 0.87). The results show no significant differences in larval RMR under light and darkness, suggesting similar larval activity levels in both conditions. The size explained most of the variability in RMR (97%), and nutritional condition was unrelated to the interindividual differences in routine metabolism. This is the first study to report the metabolic rates of Atlantic bluefin tuna larvae and discuss the challenges of performing bioenergetic studies with early life stages of scombrids.     

Nectar robbing: a common phenomenon mainly determined by accessibility constraints,nectar volume and density of energy rewards

Nectar robbers use a hole made in the perianth to extract nectar. Since robbers may modify plant fitness, they play an important role by driving evolution on floral traits, shaping population structure and influencing community dynamics. Although nectar robbing is widespread in angiosperms, the causes and ecological implications of this behaviour on large ecological scales are still unexplored. Our aim is to study the frequency of nectar robbing in plants of temperate and tropical regions and examine its association with plant traits. We characterised the levels of nectar robbing in 88 species of Mediterranean, Alpine, Antillean and Andean plant communities and identified the most important nectar robbers. We analysed associations between the levels of robbing and floral morphology, production and density of energy rewards, mechanisms of protection against nectar robbers, plant life form and geographic origin. Nectar robbing was present at all sampling sites. Within communities two patterns of robbing levels related to the diversity and specialization of robbers were detected. In most communities one plant species presented very high levels of robbing while other species had intermediate to low robbing levels. There, nectar robbers are opportunists, robbing highly rewarding plants. In the Andean community the high specialization of several co‐existing flowerpiercers produced an even pattern of robbing levels in the plant community. Plants with long flowers, abundant nectar and a high energy density are more likely to be robbed by both insects and birds. A high aggregation of the flowers within the plants and the presence of long calyxes and bracts are associated to low robbing rates by insects and to a lesser extent by birds. Besides the morphological constraints that operate on a single flower basis, nectar robbing is a phenomenon dependent upon the density of energy rewards reflecting the presence of mechanisms on higher ecological scales.     

Productive infection of Piscirickettsia salmonis in macrophages and monocyte‐like cells from rainbow trout,a possible survival strategy

Piscirickettsia salmonis is the etiologic agent of the salmonid rickettsial septicemia (SRS), an endemic disease which causes significant losses in salmon production. This intracellular bacterium is normally cultured in salmonid epithelial cell lines inducing characteristic cytopathic effects (CPEs). In this study we demonstrate that P. salmonis is able to infect, survive, replicate, and propagate in the macrophages/monocytes cell line RTS11 derived from rainbow trout spleen, without inducing the characteristic CPEs and the host cells showing the same expression levels as non‐infected control cell. On the other hand, bacteria were capable of expressing specific proteins within infected cells. Infected macrophages cease proliferation and a fraction of them detached from the plate, transform to non‐adhesive, monocyte‐like cells with proliferative activity. Productive infection of P. salmonis into salmonid macrophage/monocyte cells in culture provides an excellent model for the study of host–pathogen interactions, almost unknown in the case of P. salmonis. Our results suggest that the infection of cells from the salmonid innate immune system without inducing an important cell death response should lead to the persistence of the bacteria and consequently their dissemination to other tissues, favoring the evasion of the first line of defense against pathogens. J. Cell. Biochem. 108: 631–637, 2009. © 2009 Wiley‐Liss, Inc.     

Stabilizing proteins from sequence statistics: the interplay of conservation and correlation in triosephosphate isomerase stability

Understanding the determinants of protein stability remains one of protein science's greatest challenges. There are still no computational solutions that calculate the stability effects of even point mutations with sufficient reliability for practical use. Amino acid substitutions rarely increase the stability of native proteins; hence, large libraries and high-throughput screens or selections are needed to stabilize proteins using directed evolution. Consensus mutations have proven effective for increasing stability, but these mutations are successful only about half the time. We set out to understand why some consensus mutations fail to stabilize, and what criteria might be useful to predict stabilization more accurately. Overall, consensus mutations at more conserved positions were more likely to be stabilizing in our model, triosephosphate isomerase (TIM) from Saccharomyces cerevisiae. However, positions coupled to other sites were more likely not to stabilize upon mutation. Destabilizing mutations could be removed both by removing sites with high statistical correlations to other positions and by removing nearly invariant positions at which "hidden correlations" can occur. Application of these rules resulted in identification of stabilizing mutations in 9 out of 10 positions, and amalgamation of all predicted stabilizing positions resulted in the most stable yeast TIM variant we produced (+8 °C). In contrast, a multimutant with 14 mutations each found to stabilize TIM independently was destabilized by 2 °C. Our results are a practical extension to the consensus concept of protein stabilization, and they further suggest the importance of positional independence in the mechanism of consensus stabilization.     

Effects of citronella oil (Cymbopogon winterianus Jowitt ex Bor) on Spodoptera frugiperda (J. E. Smith) midgut and fat body

The armyworm, Spodoptera frugiperda, is the principal pest of corn in Brazil. Control is achieved primarily by synthetic insecticides, which cause problems for the agro-ecosystem. Alternative methods of control are under investigation and citronella (Cymbopogon winterianus) essential oil appears to be a promising agent. We investigated the effects of citronella oil using histological, histochemical and immunohistochemical methods. The midgut of larvae treated with citronella exhibited altered epithelium including cytoplasmic protrusions, columnar cell extrusion, pyknotic nuclei, and increased periodic acid-Schiff positive granules. Regenerative cells in the epithelium of the midgut increased in number, which facilitated subsequent regeneration of this tissue. After exposure to citronella, trophocytes, the principal cell type of the fat body, possessed enlarged vacuoles and mitotic bodies, and contained reduced amounts of glycogen, lipid, and protein. Citronella oil caused morphological changes of the midgut and reduction of stored resources in the fat body, which may adversely affect insect reproduction and survival.     

Identifying Domesticated and Wild Kañawa (<Emphasis Type="Italic">Chenopodium pallidicaule</Emphasis>) in the Archeobotanical Record of the Lake Titicaca Basin of the Andes

Here, we present new data on how to identify both the wild and domesticated forms of kañawa (Chenopodium pallidicaule Aellen) in the archeological record of Andean South America using characteristics of their seed morphology. The ability to identify both the domesticated and wild forms of kañawa is an essential step in advancing our understanding of the processes of its domestication, diversification, and the role it has played in past food systems throughout the Andes.     

Modifications of Golgi complex in chondrocytes from osteoarthrotic (OA) rat cartilage.

The status of the Golgi complex in normal vs osteoarthrotic (OA) cartilage has not yet been studied. A monoclonal antibody, MAb 58-K-9, allowed scoring of Golgi labeling intensity. In addition, ultrastructural assessment enabled us to focus on the distribution and relation between the endoplasmic reticulum (ER) and Golgi membranes. The study was performed in both normal and partially menisectomized OA-induced rat cartilage 20 and 45 days after surgery. Comparing Golgi immunolabeling intensities (mean +/- SEM) revealed a highly significant difference between normal (9.98 +/- 1.25), 20-day (2.49 +/- 0.34), and 45-day (0.82 +/- 0.22) cartilage. Moreover, chondrocytes from normal cartilage displayed 71.18% of labeling intensity in contrast to OA cartilage, in which chondrocyte labeling intensities were 24.95% (20 days) and 8.11% (45 days). OA chondrocytes appeared to display an overall reduction in Golgi labeling intensity, suggesting disruption of this organelle as the OA damage progressed. Interestingly, many 20-day OA-induced chondrocytes exhibited bubble-like Golgi immunolabeling compartmentalizing the cytoplasm, concomitant with putative apoptotic nuclear changes. At the same time, OA chondrocytes with a typical ultrastructural apoptotic pattern revealed a prominent ER gathered together with Golgi vesicles and saccules, also appearing to compartmentalize chondrocyte cytoplasm. We speculate about the role of Golgi modifications and apoptosis in OA pathogenesis.     

Large-scale cultivation ofSolanum chrysotrichum cells: Production of the antifungal saponin SC-1 in 10-I airlift bioreactors

Cells of two different cell lines:ccvx (cotyledon derived) andccvz (hypocotyl derived) ofSolanum chrysotrichum were cultivated in 10-1 airlift bioreactors for the production of the human antimycotic compound SC-1. When using 3 g l^-1 dry weight inoculum in a batch culture, higher levels of biomass were achieved with theccvx cell line (14.6 g l^-1) than withccvz (7.7 g l^-1), resulting in 23 and 12 mg g^-1 of SC-1 after 17 days in culture forccvx andccvz, respectively. The maximum productivity of SC-1 in bioreactors was 0.025 g l^-1 day^-1 after 9 days in culture. When using a draw-fill mode, the productivity increased by 60% to a value of 0.041 g l^-1 day, 4 days after 50% of the cell suspension was removed and replaced with fresh medium. This latter bioreactor system is a feasible alternative for the production of the antimycotic metabolite ofS. chrysotrichum on a large scale.Abbreviations DW Cell dry weight - FW Fresh weight - MS Murashige and Skoog (1962) medium - T _d Doubling time     

Synthesis and secretion of active lipoprotein lipase in Chinese-hamster ovary (CHO) cells.

Cultured Chinese-hamster ovary cells (CHO cells) were found to produce and secrete a lipase, which was identified as a lipoprotein lipase by the following criteria. Its activity was stimulated by serum and apolipoprotein CII, and was inhibited by high salt concentration. The lipase bound to heparin-agarose and co-eluted with 125I-labelled bovine lipoprotein lipase in a salt gradient. A chicken antiserum to bovine lipoprotein lipase inhibited the activity and precipitated a labelled protein of the same apparent size as bovine lipoprotein lipase from media of CHO cells labelled with [35S]methionine. The lipase activity and secretion were similar in growing cells and in cells that had reached confluency. Hence, lipoprotein lipase appears to be expressed constitutively in CHO cells and is not linked to certain growth conditions, as in pre-adipocyte and macrophage cell lines. At 37 degrees C, but not at 4 degrees C, heparin increased the release of lipase to the medium 2-4-fold. This increased release occurred without depletion of cell-associated lipase activity, suggesting that heparin enhanced release of newly synthesized lipase.     

Greengenes, a chimera-checked 16S rRNA gene database and workbench compatible with ARB

A 16S rRNA gene database (http://greengenes.lbl.gov) addresses limitations of public repositories by providing chimera screening, standard alignment, and taxonomic classification using multiple published taxonomies. It was found that there is incongruent taxonomic nomenclature among curators even at the phylum level. Putative chimeras were identified in 3% of environmental sequences and in 0.2% of records derived from isolates. Environmental sequences were classified into 100 phylum-level lineages in the Archaea and Bacteria.