Novel bacteriocinogenic Enterococcus hirae and Pediococcus pentosaceus strains with antilisterial activity isolated from Brazilian artisanal cheese

We isolated and characterized bacteriocin producers Enterococcus hirae ST57ACC and Pediococcus pentosaceus ST65ACC from raw milk artisanal cheeses. Their bacteriocins were tolerant to temperatures from 4°C to 100°C and under sterilization conditions (121°C for 15 min). Additionally, the tested bacteriocins remained active after being exposed to pH 2.0 to 10.0 for 2 h. The activity of the bacteriocins was affected by proteolytic enzymes but remained stable after treatment with EDTA, sodium dodecyl sulfate, NaCl, skim milk, and Tween 80. Cell-free supernatants were capable of inhibiting Listeria innocua and several strains of Listeria monocytogenes obtained from different sources and belonging to different serotypes. When L. monocytogenes 211 and L. monocytogenes 422 were treated with bacteriocins, growth was completely inhibited over 12 h. Cocultures of bacteriocinogenic strains and L. monocytogenes 422 in skim milk showed that E. hirae ST57ACC could control the growth of the pathogen in the matrix after 48 h. None of the selected isolates presented positive results on a screening panel for 25 bacteriocin-related genes, however, indicating that both strains might express novel bacteriocins.     

'Locked-on' and 'locked-off' signal transduction mutations in the periplasmic domain of the Escherichia coli NarQ and NarX sensors affect nitrate- and nitrite-dependent regulation by NarL and NarP

The Escherichia coli NarX, NarQ, NarL and NarP proteins comprise a two-component regulatory system that controls the expression of many anaerobic electron-transport and fermentation-related genes in response to nitrate and nitrite. Either of the two sensor-transmitter proteins, NarX and NarQ, can activate the response-regulator proteins, NarL and NarP, which in turn are able to bind at their respective DNA regulatory sites to modulate gene expression. NarX contains a conserved 17 amino acid sequence, designated the 'P-box' element, that is essential for nitrate sensing. In this study we characterize narQ mutants that also confer altered nitrate control of NarL-dependent nitrate reductase (narGHJI) and fumarate reductase (frdABCD) gene expression. While some narQ mutations cause the constitutive activation or repression of reporter-gene expression even when the cells are grown in the absence of the nitrate signal (i.e. a 'locked-on' phenotype), other mutations abolish nitrate-dependent control (i.e. a 'locked-off' phenotype). Interestingly the narQ (A42-->T) and narQ (R50-->Q) mutations along with the analogous narX18 (A46-->T) and narX902 (R54-->E) mutations also confer a 'locked-on' or a 'locked-off' phenotype in response to nitrite, the second environmental signal detected by NarQ and NarX. Furthermore, these narQ and narX mutations also affect NarP-dependent gene regulation of nitrite reductase (nrfABCDEFG) and aeg-46.5 gene expression in response to nitrite. We therefore propose that the NarQ sensor-transmitter protein also detects nitrate and nitrite in the periplasmic space via its periplasmic domain. A signal transduction model, which we previously proposed for NarX, is now extended to NarQ, in which a nitrate- or nitrite-detection event in the periplasmic region of the cell is followed by a signal transduction event through the inner membrane to the cytoplasmic domain of NarQ and NarX proteins to modulate their protein kinase/phosphatase activities.     

EFFECT OF SURFACTANT CONCENTRATION ON GAS HOLDUP IN A BUBBLE COLUMN WITH AN ORGANIC LIQUID

This work presents an experimental analysis of the effect of the addition of a surfactant on gas holdup in a bubble column with an organic liquid phase. For the system considered, the addition of surfactant increases the gas holdup by increasing the volume of foam within the column. The surfactant concentration has negligible effect on the intrinsic gas holdup of both the bubbling and foaming regions.     

Bidirectional connectivity via fish ladders in a large Neotropical river

The conservation of migratory fish species worldwide has been threatened by the loss of longitudinal connectivity caused by dams intercepting large rivers. One environmental management strategy for reestablishing connectivity is providing passage through fish ladders. However, ladders in Neotropical rivers have been described as ascending one‐way routes. We analysed the movements of Prochilodus lineatus through a fish ladder at a large dam—Porto Primavera—in the heavily impounded Upper Paraná River, Brazil, to determine whether the ladder connected habitats downstream and upstream of the dam, in both directions. A total of 1,419 specimens of P. lineatus were PIT‐tagged above and below the dam, and continuously monitored for 4 years. We documented bidirectional movements of P. lineatus through the fish ladder. Many individuals repeated these movements annually; one individual as many as six times. It was estimated that the cumulative probability that P. lineatus would return from downstream after descending through the ladder was 0.38, 0.50, and 0.56 in 1, 2, and 3 years, respectively. Correspondingly, return probabilities from upstream were 0.15, 0.22, and 0.26 in 1–3 years, respectively. Although return probabilities from upstream were roughly half, our results suggest the Porto Primavera fish ladder contributes to habitat connectivity, bidirectional passage, and preservation of P. lineatus. These results deviate from the perception that fishways are ineffective in Neotropical rivers. We suggest that fishways can restore the bidirectional connectivity denied to some Neotropical species, and until the services of dams are no longer needed, environmental management through fish ladders could continue to be considered as an integral part of broader conservation strategy designed to preserve native fauna.     

Differential effects of human serum and cells on the growth of Plasmodium falciparum adapted to serum-free in vitro culture conditions

A single Plasmodium falciparum isolate was adapted for growth in serum-free culture medium. The parasitemia increased from 0.5% to 20% on day 7 after thawing. The asexual forms of the parasites appeared morphologically normal and pigment formation was comparable with that seen under standard conditions with serum present. Parasites were coincubated in 96-well plates with serum, peripheral blood mononuclear cells (PBMC), and PBMC in the presence of autologous serum from healthy non-immune individuals (n = 12), healthy semi-immune individuals (n = 12), and malaria patients (n = 7). Growth was monitored for six days. The concentration of interleukin-6 and interferon-gamma (IFN-gamma) in supernatants from the continuous cultures were measured by a bioassay and an enzyme-amplified sensitivity immunoassay. The results of this study showed that parasites cultured in serum-free medium in the presence of PBMC develop more rapidly, particularly with cells from malaria patients, compared with parasites cultured alone. The growth of parasites was different if 10% autologous serum was added to the culture. Parasite growth with sera from acutely infected individuals was similar with that with sera from aparasitemic, nonimmune individuals, and both supported significantly higher parasite growth over the six-day culture period compared with sera from the uninfected semi-immune individuals. Production of IFN-gamma by cells from nonimmune individuals and malaria patients was higher when cultures did not contain autologous serum. Nonimmune donor cells produced high amounts of IFN-gamma, but cells from the semi-immune donors produced little of this cytokine. There was no marked inhibition of parasite growth with any combination of serum and cells over six days of culture. A difference between the groups was observed after two days of culture, when growth with cells and serum from the uninfected, semi-immune group was significantly lower than that from the nonimmune group, but this was not subsequently sustained. The results of the study show that continuous cultivation of P. falciparum in serum-free medium provides a novel in vitro model to study mechanisms of the interplay between components of the human immune system and the malarial parasite, in which any possible influence of human serum is removed.     

DETERMINING THE NUMBER OF REGIMES IN MARKOV SWITCHING VAR AND VMA MODELS

We give stable finite‐order vector autoregressive moving average (p ^* ,q ^* ) representations for M‐state Markov switching second‐order stationary time series whose autocovariances satisfy a certain matrix relation. The upper bounds for p ^* and q ^* are elementary functions of the dimension K of the process, the number M of regimes, the autoregressive and moving‐average orders of the initial model. If there is no cancellation, the bounds become equalities, and this solves the identification problem. Our classes of time series include every M‐state Markov switching multi‐variate moving‐average models and autoregressive models in which the regime variable is uncorrelated with the observable. Our results include, as particular cases, those obtained by Krolzig (1997) and improve the bounds given by Zhang and Stine (2001) and Francq and Zakoïan (2001) for our classes of dynamic models. A Monte Carlo experiment and an application on foreign exchange rates complete the article. Copyright © 2013 John Wiley & Sons, Ltd.