Enhancing rotifer Brachionus calyciflorus population growth with commercial probiotics

The effect of a commercial probiotic (NanoCrusta, Altacrusta, Mexico City, Mexico) on the growth performance of Brachionus calyciflorus Pallas, 1766, was evaluated. In a first approach, probiotics were supplied in four densities (2.0 × 10³, 1.1 × 10⁵, 2.1 × 10⁵ and 2.1 × 10⁶ cells/ml), alone and in combination with Chlorella vulgaris (1 × 10⁶ cells/ml). The test rotifer did not grow on the probiotic alone. However, when probiotics + C. vulgaris were added, the maximum densities (D_max; ind/ml) and population growth rates (r) observed were higher. In the second experiment, probiotics were supplied at five higher densities (2.1 × 10⁶, 4.2 × 10⁶, 8.5 × 10⁶, 1.7 × 10⁷ and 3.4 × 10⁷ cells/ml) with C. vulgaris and a control treatment with only C. vulgaris (probiotic‐free). Treatments supplied with probiotics between 2.1 × 10⁶ and 1.7 × 10⁷ cells/ml showed significantly higher D_max and r than the control treatment. The results showed a positive effect of probiotic bacteria when supplied with C. vulgaris. The best outcome showed a D_max 2.16 times and an r 1.63 times higher than the density of the control treatment. Growth rates were higher in the treatments with probiotics compared to the control. We conclude that application of NanoCrusta is feasible to improve B. calyciflorus production, but the effects need to be tested in larger scales.     

Trypanocidal Action of (?)-Elatol Involves an Oxidative Stress Triggered by Mitochondria Dysfunction

Natural compounds have shown good potential for the discovery of new chemotherapeutics for the treatment of Chagas’ disease. Recently, our group reported the effective trypanocidal activity of (−)-elatol, extracted from the red macroalgae Laurencia dendroidea present in the Brazilian coast against Trypanosoma cruzi. However, the mechanism of action of this compound has remained unclear. There are only hypotheses concerning its action on mitochondrial function. Here, we further investigated the mechanisms of action of (−)-elatol on trypomastigotes of T. cruzi. For this, we evaluated some biochemical alterations in trypomastigotes treated with (−)-elatol. Our results show that (−)-elatol induced depolarization of the mitochondrial membrane, an increase in the formation of mitochondrial superoxide anion and loss of cell membrane and DNA integrity. Additionally, (−)-elatol induced formation of autophagic vacuoles and a decrease in cell volume. All together, these results suggest that the trypanocidal action of (−)-elatol involves multiple events and mitochondria might be the initial target organelle. Our hypothesis is that the mitochondrial dysfunction leads to an increase of ROS production through the electron transport chain, which affects cell membrane and DNA integrity leading to different types of parasite death.     

Expression of toll-like receptor 2 (TLR2) in porcine leukocyte subsets and tissues

Toll-like receptors (TLR) are a group of pattern recognition molecules that play a crucial role in innate immunity. TLR2 recognises a variety of microbial components leading to the development of inflammatory and immune responses. To characterise the expression and functional properties of porcine TLR2 (pTLR2), we have raised a panel of monoclonal antibodies (mAb) against this molecule. Mouse 3T3 cell transfectants expressing pTLR2 were used for immunisation of mice. The specificity of these antibodies was confirmed by their reactivity with CHO cells transfected with pTLR2 but not with pTLR4 or with non-transfected cells. Using one of these mAbs, named 1H11, pTLR2 was found on cells of the innate immune system, including monocytes, macrophages, and granulocytes, but not on peripheral blood lymphocytes. Staining of tissue sections showed that pTLR2 is also expressed on epithelial cells lining the tracheobronchial and intestinal tracts, bile ducts in the liver and renal tubules, and on the basal layer of the epidermis. This distribution is consistent with a surveillance function at entry sites, allowing for early detection of microbial invasion.     

Effect of colored plastic mulch on growth,yield and nutrient status in cucumber under shade house and open field conditions

The aim of this study was to realize whether soil mulching, with different plastic mulch colors, is a suitable practice under shade house (SH) conditions for the culture of cucumber. To do so, cucumber was cultured mulched or not with black, blue, red or white-on-black plastic films under SH, and contrasted against mulched cucumber in open field (OF). Red mulch produced the highest shoot dry weight per plant and bare soil the lowest. However, it was the white mulch which produced the highest commercial yield per plant. Contrastingly, bare soil plants produced the lowest commercial yield. SH plants two folded photosynthetic rates compared to OF plants. Mulch color mainly impacted leaf phosphorus (P) and magnesium (Mg) content while the SH affected nitrogen (K), calcium (Ca) and magnesium (Mg). Our results confirm that soil mulching, and shading positively impact the cucumber yield and quality but also show that soil mulching under SH enhances cucumber crop.     

Application of isotope dilution analysis for the evaluation of extraction conditions in the determination of total selenium and selenomethionine in yeast-based nutritional supplements

Isotope dilution analysis (IDA) has been used to quantify total selenium, total solubilized selenium, and the selenomethionine (SeMet) amount in yeast and yeast-based nutritional supplements after acid microwave digestion and different enzymatic extraction procedures. For this purpose, both a (77)Se-enriched SeMet spike, previously synthesized and characterized in our laboratory, and a (77)Se(VI) spike were used. In the analysis of the nutritional supplements, the SeMet spike was added to the sample and extracted under different conditions, and the (78)Se/(77)Se and (80)Se/(77)Se isotope ratios were measured as peak area ratios after high-performance liquid chromatography (HPLC) separation and inductively coupled plasma mass spectrometry (ICP-MS) detection. The formation of SeH(+) and mass discrimination were corrected using a natural SeMet standard injected every three samples. Similarly, total solubilized selenium was measured in the extracts after enzymatic hydrolysis using the (77)Se-enriched SeMet as a spike by direct nebulization without a chromatographic separation. To establish a mass balance, total selenium was also determined by IDA-ICP-MS on the yeast tablets after microwave digestion using (77)Se(VI) as a spike. Results showed that all enzymatic procedures tested were able to solubilize total selenium quantitatively from the solid. However, the recovery for the species SeMet, the major selenium compound detected, was seriously affected by the enzymatic procedure employed and also by the matrix composition of the supplement evaluated. For the yeast sample, SeMet recovery increased from 68 to 76% by the combined use of driselase and protease. For the nutritional supplements, the two most effective procedures appeared to be protease and driselase/protease, with a SeMet recovery ranging from 49 to 63%, depending upon the supplement evaluated. In the case of in vitro gastrointestinal enzymolysis, the results obtained showed 26-37% SeMet recovery, while the rest of selenium was solubilized as other unknown compounds (probably Se-containing peptides).     

Comparison of lignin and cellulose solubilities in ionic liquids by COSMO-RS analysis and experimental validation

In this work, the solubilities of lignin and cellulose in ionic liquids (ILs) have been analyzed and compared by COSMO-RS simulation. The excess enthalpy of the IL + lignin/cellulose mixtures is evaluated as reference property to describe the affinity of lignin and cellulose for different ionic liquids, in terms of the intermolecular interactions contributing to their mixing properties. Thus, the anion plays the main role in the dissolution process of both lignin and cellulose, in which the hydrogen bonding forces are the major interactions. The most promising anions for the cellulose and lignin dissolution are acetate, formate and chloride, whereas different cations can be employed. Computational results indicated that a rational combination of the anions and cations allows designing ILs able to dissolve with different selectivity lignin and cellulose. Some of these solutions are tested in the laboratory to validate the model employed and the regenerated solids are analyzed by FTIR spectroscopy.     

A novel application of the ecological field theory to the definition of physiographic and climatic potential areas of forest species

A new approach to the definition of physiographic and climatic potential areas for forest species, based on the ecological field theory, is outlined in this paper. The proposed formulation is tested on the Spanish juniper (Juniperus thurifera L.), using data from 883 permanent and temporary plots throughout its distribution area in the Spanish autonomous region of Castilla y León. The suitability of the territory for the species is assessed by previously studying its habitat, which in turn is analyzed through physiographic and climatic parameters. This new method is rooted in an additive index that depends on the Mahalanobis distance in the parametric space that evaluates the ecological resemblance between the studied site and each of the points defining the parametric habitat. Thereby the ecological potential of any site within the territory can be established, integrated in a geographical information systems and accordingly charted. The results are compared with those obtained with the methodology traditionally used by Spanish foresters (factorial index), showing that the overall potential area is similar in size but quite different in its distribution.     

Shiga toxin-producing Escherichia coli O157:H7 from extensive cattle of the fighting bulls breed

Shiga toxin-producing Escherichia coli (STEC) O157:H7 represents a major public health concern worldwide, with cattle recognized as their main natural reservoir. The aim of this work was to determine the prevalence and the pheno-genotypic characteristics of STEC O157:H7 in a herd with 268 cattle of the fighting bulls breed (De Lidia breed) managed under extensive conditions in the South-West of Spain. Rectal-anal swabs of all animals were collected and examined for STEC O157:H7 by performing an immunomagnetic concentration and separation procedure combined with PCR, and the resulting isolates were characterized by both phenotypic and genotypic methods. Overall, STEC O157:H7 was isolated from seven animals (2.6%) in the herd. The PCR procedure indicated that all seven isolates displayed stx₂, eae-γ1, ehxA, O157 rfbE, and fliCh7 genes. They belonged to phage types 4 (one isolate) and 42 (two isolates), and four isolates reacted with typing phages but did not conform to a recognized pattern. Among the seven isolates there were five indistinguishable PFGE patterns and other two which differed only in ?2 restriction fragments, supporting the existence of horizontal transmission among animals in the herd. The present study demonstrates that cattle managed under extensive conditions in Spain can excrete STEC O157:H7 with their faeces. To our knowledge this is the first isolation of this pathogen from De Lidia cattle.     

Artificial neural network implementation in single low-cost chip for the detection of insecticides by modeling of screen-printed enzymatic sensors response

This work presents a hardware implementation of artificial neural networks (ANNs) using a dsPIC^® microcontroller to resolve mixtures of pesticides measured by amperometric acetylcholinesterase (AChE) biosensors. The response of three biosensors with different concentrations of Chlorpyrifos Oxon (CPO) and Chlorfenvinfos (CFV) was modeled by two ANNs, which were implemented on the dsPIC^®. The performance of the ANNs was good, the prediction ability was better than 0.986 when the obtained values were compared with those expected for a set of eight external test samples, which were not used for training. This implementation is proposed to develop low-cost analytical chemical specialized tools.     

Manufacture and properties of bifidogenic saccharides derived from wood mannan

Pinus pinaster wood samples were subjected to double hydrothermal processing. The liquors coming from the second stage, containing soluble saccharides of polymeric or oligomeric nature from hemicelluloses (POHs), were subjected to membrane processing (operating in discontinuous diafiltration) for refining and fractionation. Refined POH fractions were characterized by matrix-assisted laser desorption/ionization time of flight mass spectrometry and chromatographic techniques. The most complex POH component was made up of 14 hexoses and contained 4 acetyl groups. The fermentability of purified POHs by human fecal inocula was assessed by measuring both carbon source consumption and formation of short-chain fatty acids. The bifidogenic ability of POHs was confirmed by fluorescence in situ hybridization. The stimulatory effects on the bifidobacterial population reached by POHs were of the same order as those obtained with commercial fructooligosaccharides.