Browsing by Autor "Rajal, Verónica B"

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[Comparison of nucleic acid extraction efficiency using different commercial kits and qPCR. Effect of inhibitors].
(2012) Poma, Hugo R; Davies, Carolina; Gutiérrez Cacciabue, Dolores; Mora, María C; Basombrío, Miguel Á; Rajal, Verónica B
The detection of specific nucleic acid (NA) sequences by PCR has revolutionized the biological and medical sciences. Real-time PCR (qPCR) opened up the possibility of obtaining quantitative results. NA extraction is a decisive step prior to qPCR since it may produce either the removal or co-extraction of inhibitory substances of the enzymatic reaction, which in turn affects the amplification efficiency. In the present work we compared the commercial NA extraction kits from Qiagen, Invitrogen and Macherey-Nagel, which were used to extract DNA from mice blood artificially infected with Trypanosoma cruzi and PP7 RNA, Pseudomonas aeruginosa bacteriophage, in spiked aqueous matrices. NA recovery efficiency in samples without inhibitors was similar for the three extraction kits. However, the Invitrogen kit was the only one that remained unaffected in the presence of inhibitors in the samples.
Construction of a combined soil quality indicator to assess the effect of glyphosate application.
(2019) Romano-Armada, Neli; Amoroso, María J; Rajal, Verónica B
Although the use of agrochemicals allowed increasing the crops productivity, in many cases led to soil deterioration. In this study, eight composite samples from different soils of two locations (San Martín and Anta) in Salta, Argentina, were collected and analyzed. All the samples were from loamy Entisols (0-20 cm depth) under reduced tillage without and with direct spray application of glyphosate. Twenty six variables were determined (physical, chemical, and biological soil quality indicators). From them, those of higher specificity and sensitivity to changes following glyphosate application were identified by a stepwise reduction of variables aided by statistical analysis. Samples were grouped regarding location and application of glyphosate, to identify differential effects upon variables, and glyphosate sensitive variables were selected by discarding those influenced by other factors. Thence, they were used to compose a first approximation to a combined soil quality indicator (CSQI) to assess the effect of glyphosate use in agriculture upon the soil. Overall, the set of physical variables showed the same discriminating structure as the biological set. Finally, two biological, two chemical, and two physical indicators resulted as the most specific to quality variations by the application of the herbicide, being the most sensitive the microbial biomass carbon and the (Aminomethyl)phosphonic acid concentration in soil. When these two were considered into a CSQI, it was possible to discriminate samples with the application of glyphosate (lower quality) from those without application (higher quality). To the best of our knowledge, this is the first attempt to propose a CSQI that could play an important role to prevent degradation in soils subjected to glyphosate application, as it could aid in the early detection of soil quality loss. This would provide to land managers a decision tool to let the land rest from glyphosate application, to ensure sustainable practices in agriculture.
Development of a relative risk model for drinking water regulation and design recommendations for a peri urban region of Argentina.
(2015) Rodriguez-Alvarez, María Soledad; Weir, Mark H; Pope, Joanna M; Seghezzo, Lucas; Rajal, Verónica B; Salusso, María Mónica; Moraña, Liliana B
Argentina is a developing Latin American nation that has an aim of achieving the United Nations Millennium Development Goals for potable water supplies. Their current regulations however, limit the continued development of improved potable water quality and infrastructure from a microbiological viewpoint. This is since the current regulations are focused solely to pathogenic Eschericia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa) and fecal indicators. Regions of lower socioeconomic status such as peri-urban areas are particularly at risk due to lessened financial and political ability to influence their environmental quality and infrastructure needs. Therefore, a combined microbiological sampling, analysis and quantitative microbial risk assessment (QMRA) modeling effort were engaged for a peri-urban area of Salta Argentina. Drinking water samples from home taps were analyzed and a QMRA model was developed, results of which were compared against a general 1:10,000 risk level for lack of a current Argentinian standard. This QMRA model was able to demonstrate that the current regulations were being achieved for E. coli but were less than acceptable for P. aeruginosa in some instances. Appropriate health protections are far from acceptable for Giardia for almost all water sources. Untreated water sources were sampled and analyzed then QMRA modeled as well, since a significant number of the community (∼9%) still use them for potable water supplies. For untreated water E. coli risks were near 1:10,000, however, P. aeruginosa and Giardia risks failed to be acceptable in almost all instances. The QMRA model and microbiological analyses demonstrate the need for improved regulatory efforts for the peri-urban area along with improved investment in their water infrastructure.
How long can culturable bacteria and total DNA persist in environmental waters? The role of sunlight and solid particles.
(2016) Gutiérrez-Cacciabue, Dolores; Cid, Alicia G; Rajal, Verónica B
In this work, sunlight inactivation of two indicator bacteria in freshwater, with and without solid particles, was studied and the persistence of culturable cells and total DNA was compared. Environmental water was used to prepare two matrices, with and without solid particles, which were spiked with Escherichia coli and Enterococcus faecalis. These matrices were used to prepare microcosm bags that were placed in two containers: one exposed to sunlight and the other in the dark. During one month, samples were removed from each container and detection was done by membrane filter technique and real-time PCR. Kinetic parameters were calculated to assess sunlight effect. Indicator bacteria without solid particles exposed to sunlight suffered an immediate decay (<4h) compared with the ones which were shielded from them. In addition, the survival of both bacteria with solid particles varied depending on the situation analyzed (T99 from 3 up to 60days), being always culturable E. coli more persistent than E. faecalis. On the other side, E. faecalis DNA persisted much longer than culturable cells (T99>40h in the dark with particles). In this case active cells were more prone to sunlight than total DNA and the protective effect of solid particles was also observed. Results highlight that the effects caused by the parameters which describe the behavior of culturable microorganisms and total DNA in water are different and must be included in simulation models but without forgetting that these parameters will also depend on bacterial properties, sensitizers, composition, type, and uses of the aquatic environment under assessment.
Potential of Bioremediation and PGP Traits in Streptomyces as Strategies for Bio-Reclamation of Salt-Affected Soils for Agriculture.
(2020) Romano-Armada, Neli; Yañez-Yazlle, María Florencia; Irazusta, Verónica P; Rajal, Verónica B; Moraga, Norma B
Environmental limitations influence food production and distribution, adding up to global problems like world hunger. Conditions caused by climate change require global efforts to be improved, but others like soil degradation demand local management. For many years, saline soils were not a problem; indeed, natural salinity shaped different biomes around the world. However, overall saline soils present adverse conditions for plant growth, which then translate into limitations for agriculture. Shortage on the surface of productive land, either due to depletion of arable land or to soil degradation, represents a threat to the growing worldwide population. Hence, the need to use degraded land leads scientists to think of recovery alternatives. In the case of salt-affected soils (naturally occurring or human-made), which are traditionally washed or amended with calcium salts, bio-reclamation via microbiome presents itself as an innovative and environmentally friendly option. Due to their low pathogenicity, endurance to adverse environmental conditions, and production of a wide variety of secondary metabolic compounds, members of the genus Streptomyces are good candidates for bio-reclamation of salt-affected soils. Thus, plant growth promotion and soil bioremediation strategies combine to overcome biotic and abiotic stressors, providing green management options for agriculture in the near future.
Quantification of viable protozoan parasites on leafy greens using molecular methods.
(2021) Kim, Minji; Shapiro, Karen; Rajal, Verónica B; Packham, Andrea; Aguilar, Beatriz; Rueda, Lezlie; Wuertz, Stefan
Protozoan contamination in produce is of growing importance due to their capacity to cause illnesses in consumers of fresh leafy greens. Viability assays are essential to accurately estimate health risk caused by viable parasites that contaminate food. We evaluated the efficacy of reverse transcription quantitative PCR (RT-qPCR), propidium monoazide coupled with (q)PCR, and viability staining using propidium iodide through systematic laboratory spiking experiments for selective detection of viable Cryptosporidium parvum, Giardia enterica, and Toxoplasma gondii. In the presence of only viable protozoa, the RT-qPCR assays could accurately detect two to nine (oo)cysts/g spinach (in 10 g processed). When different proportions of viable and inactivated parasite were spiked, mRNA concentrations correlated with increasing proportions of viable (oo)cysts, although low levels of false-positive mRNA signals were detectable in the presence of high amounts of inactivated protozoa. Our study demonstrated that among the methods tested, RT-qPCR performed more effectively to discriminate viable from inactivated C. parvum, G. enterica and T. gondii on spinach. This application of viability methods on leafy greens can be adopted by the produce industry and regulatory agencies charged with protection of human public health to screen leafy greens for the presence of viable protozoan pathogen contamination.
The impact of point source pollution on shallow groundwater used for human consumption in a threshold country.
(2012) Cruz, Mercedes Cecilia; Cacciabue, Dolores Gutiérrez; Gil, José F; Gamboni, Oscar; Vicente, María Soledad; Wuertz, Stefan; Gonzo, Elio; Rajal, Verónica B
Many developing and threshold countries rely on shallow groundwater wells for their water supply whilst pit latrines are used for sanitation. We employed a unified strategy involving satellite images and environmental monitoring of 16 physico-chemical and microbiological water quality parameters to identify significant land uses that can lead to unacceptable deterioration of source water, in a region with a subtropical climate and seasonally restricted torrential rainfall in Northern Argentina. Agricultural and non-agricultural sources of nitrate were illustrated in satellite images and used to assess the organic load discharged. The estimated human organic load per year was 28.5 BOD(5) tons and the N load was 7.5 tons, while for poultry farms it was 9940-BOD(5) tons and 1037-N tons, respectively. Concentrations of nitrates and organics were significantly different between seasons in well water (p values of 0.026 and 0.039, respectively). The onset of the wet season had an extraordinarily negative impact on well water due in part to the high permeability of soils made up of fine gravels and coarse sand. Discriminant analysis showed that land uses had a pronounced seasonal influence on nitrates and introduced additional microbial contamination, causing nitrification and denitrification in shallow groundwater. P-well was highly impacted by a poultry farm while S-well was affected by anthropogenic pollution and background load, as revealed by Principal Component Analysis. The application of microbial source tracking techniques is recommended to corroborate local sources of human versus animal origin.