Growth and nitrogen uptake by Salicornia europaea and Aster tripolium in nutrient conditions typical of aquaculture wastewater.

The increasing need for environmentally sound aquaculture development can, in part, be addressed by using halophytic plants in integrated multitrophic aquaculture systems (IMTA) to remove waste dissolved nitrogen (N). However, knowledge of plant ability to take up nitrogen is of foremost importance to predict plants performance in such systems. Two species, Salicornia europaea and Aster tripolium, have been identified as potential candidates for IMTA due to their salt tolerance, potential N removal capabilities and their high commercial value as an additional crop. This study investigated the growth and N uptake rates of these two species under different N supply (NH4(+), NO3(-), NH4NO3). S. europaea plants produced a lower biomass when grown in NH4(+) compared to NO3(-) or NH4NO3, while A. tripolium biomass was not affected by the form in which N was supplied. N uptake in plants incubated at different concentrations of (15)N enriched solution (up to 2 mmol l(-1)) fitted the Michaelis-Menten model. While S. europaea NH4-N maximum uptake did not differ between starved and non-starved plants, A. tripolium NH4-N uptake was higher in starved plants when supplied alone. When NO3(-) was supplied alone, NO3-N maximum uptake was lower, for both species, when the plants were not starved. Comparison of starved and non-starved plants N uptake demonstrates the need for cautious interpretation of N uptake rates across different conditions. According to the observed results, both S. europaea and A. tripolium are capable of significantly high biomass production and N removal making them potential species for inclusion in efficient IMTA.

Halophyte filter beds for treatment of saline wastewater from aquaculture.

The expansion of aquaculture and the recent development of more intensive land-based marine farms require efficient and cost-effective systems for treatment of highly nutrient-rich saline wastewater. Constructed wetlands with halophytic plants offer the potential for waste-stream treatment combined with production of valuable secondary plant crops. Pilot wetland filter beds, constructed in triplicate and planted with the saltmarsh plant Salicornia europaea, were evaluated over 88 days under commercial operating conditions on a marine fish and shrimp farm. Nitrogen waste was primarily in the form of dissolved inorganic nitrogen (TDIN) and was removed by 98.2 ± 2.2% under ambient loadings of 109-383 µmol l(-1). There was a linear relationship between TDIN uptake and loading over the range of inputs tested. At peak loadings of up to 8185 ± 590 µmol l(-1) (equivalent to 600 mmol N m(-2) d(-1)), the filter beds removed between 30 and 58% (250 mmol N m(-2) d(-1)) of influent TDIN. Influent dissolved inorganic phosphorus levels ranged from 34 to 90 µmol l(-1), with 36-89% reduction under routine operations. Dissolved organic nitrogen (DON) loadings were lower (11-144 µmol l(-1)), and between 23 and 69% of influent DON was removed during routine operation, with no significant removal of DON under high TDIN loading. Over the 88-day study, cumulative nitrogen removal was 1.28 mol m(-2), of which 1.09 mol m(-2) was retained in plant tissue, with plant uptake ranging from 2.4 to 27.0 mmol N g(-1) dry weight d(-1). The results demonstrate the effectiveness of N and P removal from wastewater from land-based intensive marine aquaculture farms by constructed wetlands planted with S. europaea.

Inhibitory activity of cheese whey fermented with kefir grains.

We investigated the chemical and microbiological compositions of three types of whey to be used for kefir fermentation as well as the inhibitory capacity of their subsequent fermentation products against 100 Salmonella sp. and 100 Escherichia coli pathogenic isolates. All the wheys after fermentation with 10% (wt/vol) kefir grains showed inhibition against all 200 isolates. The content of lactic acid bacteria in fermented whey ranged from 1.04 × 10(7) to 1.17 × 10(7) CFU/ml and the level of yeasts from 2.05 × 10(6) to 4.23 × 10(6) CFU/ml. The main changes in the chemical composition during fermentation were a decrease in lactose content by 41 to 48% along with a corresponding lactic acid production to a final level of 0.84 to 1.20% of the total reaction products. The MIC was a 30% dilution of the fermentation products for most of the isolates, while the MBC varied between 40 and 70%, depending on the isolate. The pathogenic isolates Salmonella enterica serovar Enteritidis 2713 and E. coli 2710 in the fermented whey lost their viability after 2 to 7 h of incubation. When pathogens were deliberately inoculated into whey before fermentation, the CFU were reduced by 2 log cycles for E. coli and 4 log cycles for Salmonella sp. after 24 h of incubation. The inhibition was mainly related to lactic acid production. This work demonstrated the possibility of using kefir grains to ferment an industrial by-product in order to obtain a natural acidic preparation with strong bacterial inhibitory properties that also contains potentially probiotic microorganisms.

Isolation and characterization of bacteriophages for avian pathogenic E. coli strains.

AIMS: To isolate and characterize bacteriophages, and to evaluate its lytic performance against avian pathogenic Escherichia coli (APEC) strains with high patterns of antibiotic resistance, in order to select phages for a therapeutic product to treat colibacillosis in chickens. METHODS AND RESULTS: Bacteriophages were isolated from poultry sewage and tested against 148 O-serotyped APEC strains. The morphological characterization of the bacteriophages was made by transmission electronic microscopy (TEM) observations and the genetic comparison between bacteriophages DNA was performed by restriction fragment length polymorphism (RFLP) patterns. Results showed that 70.5% of the tested E. coli strains were sensitive to a combination of three of the five isolated phages, that seemed to be virulent and taxonomically belong to the Caudovirales order. Two of them look like 16-19, T4-like phages (Myoviridae) and the third is a T1-like phage and belongs to Syphoviridae family. All of them are genetically different. CONCLUSIONS: It was possible to obtain a combination of three different lytic bacteriophages with broad lytic spectra against the most prevalent O-serotypes of APEC. SIGNIFICANCE AND IMPACT OF THE STUDY: Data reported in this study, presents an in vitro well studied phage product to be used as antimicrobial agent to treat colibacillosis in poultry industry.

Gene expression profiling in vLINCL CLN6-deficient fibroblasts: Insights into pathobiology.

The CLN6 vLINCL is caused by molecular defects in CLN6 gene coding for an ER resident transmembrane protein whose function is unknown. In the present study gene expression profiling of CLN6-deficient fibroblasts using cDNA microarray was undertaken in order to provide novel insights into the molecular mechanisms underlying this neurodegenerative fatal disease. Data were validated by qRT-PCR. Statistically significant alterations of expression were observed for 12 transcripts. The two most overexpressed genes, versican and tissue factor pathway inhibitor 2, are related to extracellular matrix (ECM), predicting changes in ECM-related proteins in CLN6-deficient cells. Transcript profiling also suggested alterations in signal transduction pathways, apoptosis and the immune/inflammatory response. Up-regulated genes related to steroidogenesis or signalling, and the relationship between cholesterol dynamics and glycosphingolipid sorting, led to investigation of free cholesterol and gangliosides in CLN6-deficient fibroblasts. Cholesterol accumulation in lysosomes suggests a homeostasis block as a result of CLN6p deficiency. The cholesterol imbalance may affect structure/function of caveolae and lipid rafts, disrupting signalling transduction pathways and sorting cell mechanisms. Alterations in protein/lipid intracellular trafficking would affect the composition and function of endocytic compartments, including lysosomes. Dysfunctional endosomal/lysosomal vesicles may act as one of the triggers for apoptosis and cell death, and for a secondary protective inflammatory response. In conclusion, the data reported provide novel clues into molecular pathophysiological mechanisms of CLN6-deficiency, and may also help in developing disease biomarkers and therapies for this and other neurodegenerative diseases.