Innate Predator Recognition and the Problem of Introduced Trout

Innate predator recognition typically only occurs when there is an evolutionary history between predator and prey. Predator introductions thus can pose a substantial threat to native fauna that rely heavily on inherent identification of predators. In permanent aquatic habitats prey often encounter a variety of predatory and non-predatory fish species, and the ability to distinguish between the two is essential to avoid wasted time and energy spent in unnecessary antipredatory efforts. Here, we present a study evaluating the ability of lab-reared larvae of an endangered fully aquatic salamander (hellbenders: Cryptobranchus alleganiensis ) to recognize chemical cues from native and introduced fish predators. We recorded responses of hellbender larvae to chemical stimuli from native and non-native predatory fishes, a non-predatory fish and a blank control. Eastern hellbender larvae ( C. a. alleganiensis ) significantly reduced activity in response to chemical stimuli from native predators ( Micropterus salmoides , Micropterus dolomieu , Ambloplites rupestris , Sander vitreus , and Cottus carolinae ), but responses to non-native rainbow ( Oncorhynchus mykiss ) and brown ( Salmo trutta ) trout were not significantly different from responses to the non-predatory control (redhorse sucker, Moxostoma spp.). Responses of larval Ozark hellbenders ( C. a. bishopi ) to brown trout were similar to that of the native fishes and different from the blank control, but responses to rainbow trout did not differ from the blank control. The generally weak responses of larval hellbenders to chemical cues from introduced predatory trout could lead to increased predation in the wild, which may have exacerbated the decline of hellbender populations.     

Relationship between osmotic stress-induced abscisic acid accumulation,biomass production and plant growth in drought-tolerant and -sensitive wheat cultivars

The effects of increasing osmotic stress induced by 100–400 mOsm (−0.976 MPa) polyethylene glycol (PEG 6000) were investigated in a drought-tolerant (Triticum aestivum L. cv. Mv Emese) and drought-sensitive (cv. GK élet) wheat cultivar at the three-leaf stage. During osmotic stress, the decline of the water potential (ψ _w) was more significant in the leaves, while the abscisic acid (ABA) levels of the roots increased earlier and remained higher in the sensitive than in the tolerant variety. There was an increasing gradient of ABA content toward the youngest leaves in the drought-sensitive GK élet, while more ABA accumulated in the fully developed, older leaves of the tolerant cultivar Mv Emese. In accordance with the rapid and significant accumulation of ABA, the stomatal conductance decreased earlier in the tolerant cultivar. The effect of water stress on the PSII photochemistry was pronounced only 1 week after the exposure to PEG, as indicated by the earlier decrease of the net CO₂ fixation, the effective quantum yield (Φ_PSII) and the photochemical quenching (q _P) in light-adapted samples of the tolerant variety in 400 mOsm PEG 6000. The stress treatment caused more significant reductions in these parameters toward the end of the experiment in the sensitive cultivar. In spite of small differences in the photosynthetic characteristics, the net biomass production was not significantly altered by this osmotic stress. The accumulation of ABA controlled the distribution of the biomass between the shoot and root systems under osmotic stress, and contributed to the development of stronger and deeper roots in the drought-sensitive cultivar GK élet. However, the root elongation did not correlate with the drought sensitivity of these cultivars on the basis of crop yield.     

Use of 17α-methyltestosterone to sex inverse gynogenic female rainbow trout

Oral administration of 17α-methyltestosterone (MT) was used in two experiments to sex inverse gynogenic rainbow trout. In both experiments MT was mixed into the feed in dosages of .5 mg, 1 mg, and 3 mg/kg. In Experiment 1 the treatment lasted for 300° days, 450° days, and 700° days, and in Experiment 2 for 600° days, 750° days and 938° days. Significant Chi-Square tests (P < 0.01, P < 0.001) regarding the number of phenotypic males in both experiments showed that MT dosage and treatment duration did not act independently. At about 1 year of age the highest numbers of phenotypic males were achieved when a lower dosage (.5 mg or 1 mg) was fed for a longer duration (750° days, 938° days); 100% males were found in the 1 mg-750° d and the .5 mg-938° d treatments. In the 1-year-old phenotypic males in Experiment 2, body weight and gonad weight were correlated with r = 0.60 (P < 0.001). The analyses of variance showed that body weight was affected by gonad weight group and MT dosage, while gonad weight was not affected by MT dosage or treatment duration. Data on sex distribution at maturity are available for Experiment 1. Here, too, the lower dosages of .5 mg and 1 mg/kg applied for the longer 700° day duration, as well as the 1 mg-450° day treatment, and achieved the best results of 60% phenotypic males.     

Polyphosphate kinase genes from full-scale activated sludge plants

The performance of enhanced biological phosphorus removal (EBPR) wastewater treatment processes depends on the presence of bacteria that accumulate large quantities of polyphosphate. One such group of bacteria has been identified and named Candidatus Accumulibacter phosphatis. Accumulibacter-like bacteria are abundant in many EBPR plants, but not much is known about their community or population ecology. In this study, we used the polyphosphate kinase gene (ppk1) as a high-resolution genetic marker to study population structure in activated sludge. Ppk1 genes were amplified from samples collected from full-scale wastewater treatment plants of different configurations. Clone libraries were constructed using primers targeting highly conserved regions of ppk1, to retrieve these genes from activated sludge plants that did, and did not, perform EBPR. Comparative sequence analysis revealed that ppk1 fragments were retrieved from organisms affiliated with the Accumulibacter cluster from EBPR plants but not from a plant that did not perform EBPR. A new set of more specific primers was designed and validated to amplify a 1,100 bp ppk1 fragment from Accumulibacter-like bacteria. Our results suggest that the Accumulibacter cluster has finer-scale architecture than previously revealed by 16S ribosomal RNA-based analyses. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Sugar sensing by enterocytes combines polarity, membrane bound detectors and sugar metabolism

Sugar consumption and subsequent sugar metabolism are known to regulate the expression of genes involved in intestinal sugar absorption and delivery. Here we investigate the hypothesis that sugar-sensing detectors in membranes facing the intestinal lumen or the bloodstream can also modulate intestinal sugar absorption. We used wild-type and GLUT2-null mice, to show that dietary sugars stimulate the expression of sucrase-isomaltase (SI) and L-pyruvate kinase (L-PK) by GLUT2-dependent mechanisms, whereas the expression of GLUT5 and SGLT1, did not rely on the presence of GLUT2. By providing sugar metabolites, sugar transporters, including GLUT2, fuelled a sensing pathway. In Caco2/TC7 enterocytes, we could disconnect the sensing triggered by detector from that produced by metabolism, and found that GLUT2 generated a metabolism-independent pathway to stimulate the expression of SI and L-PK. In cultured enterocytes, both apical and basolateral fructose could increase the expression of GLUT5, conversely, basolateral sugar administration could stimulate the expression of GLUT2. Finally, we located the sweet-taste receptors T1R3 and T1R2 in plasma membranes, and we measured their cognate G alpha Gustducin mRNA levels. Furthermore, we showed that a T1R3 inhibitor altered the fructose-induced expression of SGLT1, GLUT5, and L-PK. Intestinal gene expression is thus controlled by a combination of at least three sugar-signaling pathways triggered by sugar metabolites and membrane sugar receptors that, according to membrane location, determine sugar-sensing polarity. This provides a rationale for how intestine adapts sugar delivery to blood and dietary sugar provision.     

Influence of day-night and tidal cycles on phenol content and antioxidant capacity in three temperate intertidal brown seaweeds

The daily variations of phenol contents and antioxidant capacities were surveyed in a 32-h field experiment in three temperate brown seaweeds belonging to Fucales - Pelvetia canaliculata, Ascophyllum nodosum and Bifurcaria bifurcata - living at different intertidal levels — high-, mid- and low-tide level, respectively. Phenolic compounds of brown seaweeds are secondary metabolites involved in many different protection mechanisms, as for example against grazer and pathogen attack as well as UV damage. This study was thus aimed at understanding the influence of both day/night and tidal cycles on the brown seaweed phenol pool with respect to their bathymetric level on the shore. These cycles affect the quantity and quality of light received by intertidal seaweeds (protection via the water layer during immersion), and the photoprotective role of phlorotannins was thus evaluated. Phenol levels and antioxidant capacities were monitored every hour during a tidal cycle and a half at the equinox spring tide, in March 2003. The three species contained rather high phenol levels, i.e. about 3, 6 and 4% DW in P. canaliculata, A. nodosum and B. bifurcata, respectively. Antioxidant capacities globally paralleled phenol contents in the three species under study. Moreover, the measured antioxidant capacities and the phenol contents of the extracts were significantly and positively correlated in the three species. Significant effect of neither cycles nor their interaction was observed in any species despite a trend to follow day/night alternation for P. canaliculata, and emersion/immersion cycle for A. nodosum. No trend was observed for the third species. However, significant correlations between phenol levels of P. canaliculata and A. nodosum and measured air-temperature were found. Our results suggested an effect of aerial, aquatic and both conditions on the phenolic pool of P. canaliculata, B. bifurcata and A. nodosum, respectively, and a fast evolution of the phenolic pool on a day time scale.     

Lipophosphoramidates as lipidic part of lipospermines for gene delivery

The DNA compacting properties of polyamines (especially spermine) are well-known, hence the use of spermine as the cationic part in several synthetic DNA carriers. Here, we describe the synthesis of modified spermines, with a "lipophosphoramidate" as the lipidic part, and their use for efficient in vitro transfection. Physicochemical measurements (particle size, zeta potentials, pKa determination) and gel retardation assays were also performed. Theoretical membrane-disrupting ability was established by FRET. Taken together, our results indicate that lipophosphoramidates constitute an interesting alternative to "classical" lipidic parts of cationic lipids used as DNA carriers.     

Rescue and production of vaccine and therapeutic adenovirus vectors expressing inhibitory transgenes

Expression of certain transgenes from an adenovirus vector can be deleterious to its own replication. This can result in the inhibition of virus rescue, reduced viral yields, or, in the worst case, make it impossible to construct a vector expressing the inhibiting transgene product. A gene regulation system based on the tet operon was used to allow the rescue and efficient growth of adenovectors that express transgenes to high levels. A key advantage to this system is that repression of transgene expression is mediated by the packaging cell line, thus, expression of regulatory products from the adenovector are not required. This provides a simple, broadly applicable system wherein transgene repression is constitutive during vector rescue and growth and there is no effect on adenovector-mediated expression of gene products in transduced cells. Several high-level expression vectors based on first- and second-generation adenovectors were rescued and produced to high titer that otherwise could not be grown. Yields of adenovectors expressing inhibitory transgene products were increased, and the overgrowth of cultures by adenovectors with nonfunctional expression cassettes was prevented. The gene regulation system is a significant advancement for the development of adenovirus vectors for vaccine and other gene transfer applications.