A controlled aquarium system and approach to study the role of sponge-bacteria interactions using Aplysilla rosea and Vibrio natriegens.

Sponge-bacteria interactions are very important due to their ecological and biological significance. To understand the impact of interactions between sponges and bacteria (both associated with and external to sponges) on sponge-associated microbial diversity, sponge metabolite profiles and bioactivity, we used a controlled aquarium system and designed an experimental approach that allows the study of sponge-bacteria interactions in a well-defined manner. To test the feasibility of this approach, this system was used to study the interaction between a sponge Aplysilla rosea and a marine bacterium commonly found in seawater, Vibrio natriegens. Sponge explants were exposed to V. natriegens, at 5 × 106 cfu/ml, and changes were monitored for 48 hours. Pyro-sequencing revealed significant shifts in microbial communities associated with the sponges after 24 to 48 hours. Both the control (sponge only without added bacteria) and Vibrio-exposed sponges showed a distinct shift in bacterial diversity and abundance with time. Vibrio exposure significantly increased bacterial diversity, the abundance of a number of taxa compared to control sponges. The result experimentally supports the notion of dynamic and concerted responses by the sponge when interacting with a bacterium, and demonstrates the feasibility of using this controlled aquarium system for the study of sponge-bacteria interactions.

Characterization of amniotic fluid of Dohne Merino ewes (Ovis aries) and its possible role in neonatal recognition.

Amniotic fluid (AF) was investigated as a possible source of the neonatal recognition cue that plays a crucial role in ewe-lamb bonding in sheep. A total of 70 of the 133 volatile organic compounds (VOCs), previously identified in the cranial wool of neonatal Dohne Merino lambs, was also identified in AF collected individually from neonatal twins. As in the case of the wool of neonatal lambs, the VOC profiles (GC-MS) of the AF of twins were remarkably similar. However, the VOC profiles of the AF differed from that of the wool VOCs of the same lambs. The VOCs that best represent the total variance in the AF and wool of neonatal lambs were not identified as ligands of the AF proteins. These observations suggest that it is unlikely that the neonatal recognition cue of sheep could be a maternal label derived from the AF in which a lamb is born, and that the neonatal recognition cue is probably produced by the lamb. Taking cognizance of the temporal changes that take place in the quantitative composition of the cranial wool VOCs, we hypothesize that components of the neonatal recognition cue are already produced by the lamb well before its birth and that the quantitative composition in which these volatiles are released into the AF does not correspond to the composition of the recognition cue of the lamb at its birth. When grooming the newborn lamb, its mother removes AF containing incorrect chemical information from her lamb's body and this affords her enough time to learn its personal recognition cue.

Reuteran and levan as carbohydrate sinks in transgenic sugarcane.

The present study reports the effect of high molecular weight bacterial fructan (levan) and glucan (reuteran) on growth and carbohydrate partitioning in transgenic sugarcane plants. These biopolymers are products of bacterial glycosyltransferases, enzymes that catalyze the polymerization of glucose or fructose residues from sucrose. Constructs, targeted to different subcellular compartments (cell wall and cytosol) and driven by the Cauliflower mosaic virus-35S: maize-ubiquitin promoter, were introduced into sugarcane by biolistic transformation. Polysaccharide accumulation severely affected growth of callus suspension cultures. Regeneration of embryonic callus tissue into plants proved problematic for cell wall-targeted lines. When targeted to the cytosol, only plants with relative low levels of biopolymer accumulation survived. In internodal stalk tissue that accumulate reuteran (max 0.03 mg/g FW), sucrose content (ca 60 mg/g FW) was not affected, while starch content (<0.4 mg/g FW) was increased up to four times. Total carbohydrate content was not significantly altered. On the other hand, starch and sucrose levels were significantly reduced in plants accumulating levan (max 0.01 mg/g FW). Heterologous expression resulted in a reduction in total carbohydrate assimilation rather than a simple diversion by competition for substrate.

The role of bile after Roux-en-Y gastric bypass in promoting weight loss and improving glycaemic control.

Gastric bypass leads to the remission of type 2 diabetes independently of weight loss. Our hypothesis is that changes in bile flow due to the altered anatomy may partly explain the metabolic outcomes of the operation. We prospectively studied 12 patients undergoing gastric bypass and six patients undergoing gastric banding over a 6-wk period. Plasma fibroblast growth factor (FGF)19, stimulated by bile acid absorption in the terminal ileum, and plasma bile acids were measured. In canine and rodent models, we investigated changes in the gut hormone response after altered bile flow. FGF19 and total plasma bile acids levels increased after gastric bypass compared with no change after gastric banding. In the canine model, both food and bile, on their own, stimulated satiety gut hormone responses. However, when combined, the response was doubled. In rats, drainage of endogenous bile into the terminal ileum was associated with an enhanced satiety gut hormone response, reduced food intake, and lower body weight. In conclusion, after gastric bypass, bile flow is altered, leading to increased plasma bile acids, FGF19, incretin. and satiety gut hormone concentrations. Elucidating the mechanism of action of gastric bypass surgery may lead to novel treatments for type 2 diabetes.

Manipulation of L-ascorbic acid biosynthesis pathways in Solanum lycopersicum: elevated GDP-mannose pyrophosphorylase activity enhances L-ascorbate levels in red fruit.

Ascorbate (AsA) plays a fundamental role in redox homeostasis in plants and animals, primarily by scavenging reactive oxygen species. Three genes, representing diverse steps putatively involved in plant AsA biosynthesis pathways, were cloned and independently expressed in Solanum lycopersicum (tomato) under the control of the CaMV 35S promoter. Yeast-derived GDP-mannose pyrophosphorylase (GMPase) and arabinono-1,4-lactone oxidase (ALO), as well as myo-inositol oxygenase 2 (MIOX2) from Arabidopsis thaliana, were targeted. Increases in GMPase activity were concomitant with increased AsA levels of up to 70% in leaves, 50% in green fruit, and 35% in red fruit. Expression of ALO significantly pulled biosynthetic flux towards AsA in leaves and green fruit by up to 54 and 25%, respectively. Changes in AsA content in plants transcribing the MIOX2 gene were inconsistent in different tissue. On the other hand, MIOX activity was strongly correlated with cell wall uronic acid levels, suggesting that MIOX may be a useful tool for the manipulation of cell wall composition. In conclusion, the Smirnoff-Wheeler pathway showed great promise as a target for biotechnological manipulation of ascorbate levels in tomato.

Olfactory cue mediated neonatal recognition in sheep, Ovis aries.

The strong bond between ewe and lamb formed shortly after parturition is an important factor in lamb survival. Evidence exists that a ewe can distinguish her lamb by its unique smell, but the constituents of such a putative olfactory cue have not yet been identified. We have now identified 133 volatile organic compounds associated with the wool of Döhne Merino lambs that we presume may be involved in neonatal recognition. Quantitative analysis and comparison of odor profiles of the twins of 16 ewes (9.69% sample group) of a flock of 165 twin-bearing ewes revealed that the wool volatiles of twins are qualitatively and quantitatively similar, but differ from those of other twins or non-twin lambs in the flock. The 88 constituents present in at least 20% of the analyzed wool samples were considered as variables for multivariate analysis. A P-value < 0.001 was calculated, indicating that the pairing of twins according to the qualitative and quantitative composition of the wool was significant. Bioassays carried out during the lambing seasons of 2009 and 2010 confirmed the previously established role of lamb odor in ewe-lamb recognition. However, when alien lambs were dressed in jackets sprayed with synthetic mixtures formulated to match the chemical composition of the effluvia of the ewes' own lambs, ewes rejected the aliens. This is possibly because the VOCs were not released in quantitative ratios sufficiently accurate to emulate the odor of the ewes' own lambs.

Exopolysaccharide production by lactose-hydrolyzing bacteria isolated from traditionally fermented milk.

With increasing consumer demands for safer, healthier and more natural products, bacterially produced exopolysaccharides (EPSs) are becoming a viable option as an additive in designer-type foods. Fresh milk samples from cattle and sheep were collected from informal settlements in South Africa. After a three day incubation period at 25 degrees C, 550 bacterial strains were isolated and evaluated for EPS production from lactose as sole carbon source. Strains producing EPS on lactose were identified to species level with 16S rRNA gene sequencing and encompass 11 Gram-positive and 6 Gram-negative bacteria. EPS production was assigned for the first time to members of the species Staphylococcus hominis and Enterococcus lactis, and also to apparently novel species of the genera Sphingomonas and Acinetobacter. The polymers consisted mainly out of galactose and glucose, while a few isolates also incorporated rhamnose. Isolates produced diverse biopolymers as seen by significant differences in monomer ratios.