Evaluating the potential use of a dairy industry residue to induce denitrification in polluted water bodies: a flow-through experiment

Improving the effectiveness and economics of strategies to remediate groundwater nitrate pollution is a matter of concern. In this context, the addition of whey into aquifers could provide a feasible solution to attenuate nitrate contamination by inducing heterotrophic denitrification, while recycling an industry residue. Before its application, the efficacy of the treatment must be studied at laboratory-scale to optimize the application strategy in order to avoid the generation of harmful intermediate compounds. To do this, a flow-through denitrification experiment using whey as organic C source was performed, and different C/N ratios and injection periodicities were tested. The collected samples were analyzed to determine the chemical and isotopic composition of N and C compounds. The results proved that whey could promote denitrification. Nitrate was completely removed when using either a 3.0 or 2.0 C/N ratio. However, daily injection with C/N ratios from 1.25 to 1.5 seemed advantageous, since this strategy decreased nitrate concentration to values below the threshold for water consumption while avoiding nitrite accumulation and whey release with the outflow. The isotopic results confirmed that nitrate attenuation was due to denitrification and that the production of DIC was related to bacterial whey oxidation. Furthermore, the isotopic data suggested that when denitrification was not complete, the outflow could present a mix of denitrified and nondenitrified water. The calculated isotopic fractionation values (ε15NNO3/N2 and ε18ONO3/N2) might be applied in the future to quantify the efficiency of the bioremediation treatments by whey application at field-scale

In vivo potential of recombinant granulysin against human melanoma

9-kDa granulysin is a protein expressed into the granules of human cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. It has been shown to exert cytolysis on microbes and tumors. We showed previously that 9-kDa granulysin exerted cell death by apoptosis in vitro on hematological tumor cell lines and also on cells from B-cell chronic lymphocytic leukemia (B-CLL) patients. In addition, we have shown the anti-tumor efficiency of granulysin as a single agent in two in vivo models of human tumor development in athymic mice, the MDA-MB-231 mammary adenocarcinoma and the NCI-H929 multiple myeloma, without signs of overt secondary effects by itself. In this work, we have tested recombinant 9-kDa granulysin in an in vivo and especially aggressive model of melanoma development, xenografted UACC62 cells in athymic mice. Recombinant granulysin was administered once UACC62-derived tumors were detectable and it substantially retarded the in vivo development of this aggressive tumor. We could also detect apoptosis induction and increased NK cell infiltration inside granulysin-treated tumor tissues. These observations are especially interesting given the possibility of treating melanoma by intra-tumor injection

Nitrate and Nitrite Attenuation by Fe(II) Minerals: Biotic and Abiotic Reactions

Nitrate (NO3-) pollution of groundwaterhas become a relevant issue and anenvironmental priority as it is related toecological and human health problems(Rivett et al. 2008) and its concentration is still above the threshold limit of 50mg/L in many areas (Nitrate Directive, 91/676/EEC). Contamination sources of NO3 - are linked to extensive use of fertilizers, inappropriate placement of animal waste and spills from septic system effluents

Conjugation of the 9-kDa isoform of Granulysin with liposomes potentiates its cytotoxicity

Nine kDa granulysin (GRNLY) is a human cytolytic protein secreted by cytotoxic T lymphocytes (CTL) and NK cells of the immune system whose demonstrated physiological function is the elimination of bacteria and parasites. In previous studies by our group, the anti-tumor capacity of recombinant granulysin was demonstrated, both in vitro and in vivo. In the present work, we developed lipid nanoparticles whose surfaces can bind recombinant granulysin through the formation of a complex of coordination between the histidine tail of the protein and Ni2+ provided by a chelating lipid in the liposome composition and termed them LUV-GRNLY, for granulysin-bound large unilamellar vesicles. The objective of this formulation is to increase the granulysin concentration at the site of contact with the target cell and to increase the cytotoxicity of the administered dose. The results obtained in this work indicate that recombinant granulysin binds to the surface of the liposome with high efficiency and that its cytotoxicity is significantly increased when it is in association with liposomes. In addition, it has been demonstrated that the main mechanism of death induced by both granulysin and LUV-GRNLY is apoptosis. Jurkat-shBak cells are resistant to GRNLY and also to LUV-GRNLY, showing that LUV-GRNLY uses the mitochondrial apoptotic pathway to induce cell death. On the other hand, we show that LUV-GRNLY induces the expression of the pro-apoptotic members of the Bcl-2 family Bim and especially PUMA, although it also induced the expression of anti-apoptotic Bcl-xL. In conclusion, we demonstrate that binding of GRNLY to the surfaces of liposomes clearly augments its cytotoxic potential, with cell death executed mainly by the mitochondrial apoptotic pathway

Characterisation of the natural attenuation of chromium contamination in the presence of nitrate using isotopic methods. A case study from the Matanza-Riachuelo river basin, Argentina

The groundwater contamination by hexavalent chromium (Cr(VI)) in a site of the Matanza-Riachuelo River basin (MRB), Argentina, has been evaluated by determining the processes that control the natural mobility and attenuation of Cr(VI) in the presence of high nitrate (NO3−) contents. The groundwater Cr(VI) concentrations ranged between 1.9E-5 mM and 0.04 mM, while the NO3− concentrations ranged between 0.5 mM and 3.9 mM

Feasibility of using rural waste products to increase the denitrification efficiency in a surface flow constructed wetland

A surface flow constructed wetland (CW) was set in the Lerma gully to decrease nitrate (NO3−) pollution from agricultural runoff water. The water flow rate and NO3− concentration were monitored at the inlet and the outlet, and sampling campaigns were performed which consisted of collecting six water samples along the CW flow line. After two years of operation, the NO3− attenuation was limited at a flow rate of ~2.5 L/s and became negligible at ~5.5 L/s. The present work aimed to assess the feasibility of using rural waste products (wheat hay, corn stubble, and animal compost) to induce denitrification in the CW, to assess the effect of temperature on this process, and to trace the efficiency of the treatment by using isotopic tools. In the first stage, microcosm experiments were performed. Afterwards, the selected waste material was applied in the CW, and the treatment efficiency was evaluated by means of a chemical and isotopic characterization and using the isotopic fractionation (ε) values calculated from laboratory experiments to avoid field-scale interference. The microcosms results showed that the stubble was the most appropriate material for application in the CW, but the denitrification rate was found to decrease with temperature. In the CW, biostimulation in autumn-winter promoted NO3− attenuation between two weeks and one month (a reduction in NO3− between 1.2 and 1.5 mM was achieved). After the biostimulation in spring-summer, the attenuation was maintained for approximately three months (NO3− reduction between 0.1 and 1.5 mM). The ε15NNO3/N2 and ε18ONO3/N2 values obtained from the laboratory experiments allowed to estimate the induced denitrification percentage. At an approximate average flow rate of 16 L/s, at least 60% of NO3− attenuation was achieved in the CW. The field samples exhibited a slope of 1.0 for δ18O-NO3− versus δ15N-NO3−, similar to those of the laboratory experiments (0.9-1.2). Plant uptake seemed to play a minor role in NO3− attenuation in the CW. Hence, the application of stubble in the CW allowed the removal of large amounts of NO3− from the Lerma gully, especially when applied during the warm months, but its efficacy was limited to a short time period (up to three months)

Use of nitrogen and oxygen isotopes of dissolved nitrate to trace field-scale induced denitrification efficiency throughout an in-situ groundwater remediation strategy

In the framework of the Life+ InSiTrate project, a pilot-plant was established to demonstrate the viability of inducing in-situ heterotrophic denitrification to remediate nitrate (NO3−)-polluted groundwater. Two injection wells supplied acetic acid by pulses to an alluvial aquifer for 22 months. The monitoring was performed by regular sampling at three piezometers and two wells located downstream. In the present work, the pilot-plant monitoring samples were used to test the usefulness of the isotopic tools to evaluate the efficiency of the treatment. The laboratory microcosm experiments determined an isotopic fractionation (ε) for N-NO3− of −12.6 and for O-NO3− of −13.3 . These ε15NNO3/N2 and ε18ONO3/N2 values were modelled by using a Rayleigh distillation equation to estimate the percentage of the induced denitrification at the pilot-plant while avoiding a possible interference from dilution due to non-polluted water inputs. In some of the field samples, the induced NO3− reduction was higher than 50% with respect to the background concentration. The field samples showed a reduced slope between δ18O-NO3− and δ15N-NO3− (0.7) compared to the laboratory experiments (1.1). This finding was attributed to the reoxidation of NO2− to NO3− during the treatment. The NO3− isotopic characterization also permitted the recognition of a mixture between the denitrified and partially or non-denitrified groundwater in one of the sampling points. Therefore, the isotopic tools demonstrated usefulness in assessing the implementation of the field-scale induced denitrification strategy

Scanning Electron Microscope Study of Wohlfahrtia magnifica (Schiner, 1862) (Diptera: Sarcophagidae) I. Structures with Parasitic and Possible Taxonomic Meaning

The larval development of Wohlfahrtia magnifica (the most important dipteran causing sheep myiasis in the Palearctic region) has been studied by means of scanning electron microscopy. The ultrastructure of mouth-hooks, oral ridges, labial lobes, body spines and anterior and posterior peritremes is described for the first time. Their possible adaptations to a parasitic lifeway are also discussed. Thus, the use of new structures in the Sarcophagidae taxonomy is proposed from the point of view of their ultrastructure and adaptative morphology