National Agricultural Research Institute INIA

Colonia del Sacramento, Uruguay

National Agricultural Research Institute INIA

Colonia del Sacramento, Uruguay
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Harriet J.,Direccion Of Laboratorios Veterinarios | Campa J.P.,Direccion Of Laboratorios Veterinarios | Grajales M.,Apicola Integral Las Piedras | Lheritier C.,Asociacion de Exportadores de Miel | And 3 more authors.
Chemosphere | Year: 2017

Over the last decade, Uruguay has expanded and intensified its rainfed crop production. This process has affected beekeeping in several ways: for example, by reducing the space available. This has increased the density of apiaries, the risk of varroosis and acaricide use. Additionally, the dominance of no-tillage crops has increased the frequencies of application and of loads of pesticides in regions where such crops share the land with beekeeping and honey production. Therefore, the exposure of bees to xenobiotics (agricultural pesticides and veterinary products) has increased in line with pollution of hives and their products. To document pollution from hive exposure to pesticides, we surveyed the presence of 30 xenobiotics normally used in Uruguay, in recycled beeswax (RB) and in honey cappings (HC) from the main Uruguayan beekeeping regions. There was contamination of all the analyzed samples (RB and HC) with the herbicide atrazine at a range of 1–2 ng g−1. At least three or four additional xenobiotics were detected: insecticides (chlorpyrifos-ethyl and thiacloprid); fungicides (azoxystrobin and tebuconazole); and veterinary products (coumaphos, ethion, and tau-fluvalinate). The frequency of detection of chlorpyrifos-ethyl and coumaphos in RB samples was higher than in those of HC. Moreover, the concentrations of azoxystrobin, coumaphos, and tebuconazole in RB samples were higher than in HC samples. Therefore, we suggest the use of HC to produce recycled printed beeswax films for use in hives to minimize pollution transfer. © 2017 Elsevier Ltd


Albicette M.M.,National Agricultural Research Institute INIA | Leoni C.,National Agricultural Research Institute INIA | Ruggia A.,National Agricultural Research Institute INIA | Scarlato S.,National Agricultural Research Institute INIA | And 3 more authors.
Outlook on Agriculture | Year: 2017

There are opportunities to improve livestock family farms (LFFs) sustainability in Uruguay by using the co-innovation approach to change management practices and incorporate technologies. To harness these opportunities, between 2012 and 2015, a research project was implemented in Eastern Uruguay, where three simultaneous processes occurred at three levels: farm, region, and research team. At farm level, the work was carried out in seven LFF as case studies following three steps: (i) characterization and diagnosis; (ii) redesign; and (iii) implementation, monitoring, and evaluation. At the regional level, a participatory approach to planning, monitoring, and evaluating the project’s progress with regional stakeholders was adopted. At the team level, a Participatory Action Research approach was used. Consensus on the objectives and methods allowed for combined knowledge to solve practice-oriented problems. The 3-year project demonstrated the effectiveness in improving LFF sustainability, opened a learning space with stakeholders, and utilized a co-innovation model to improve rural development outcomes. © The Author(s) 2017.


Arruabarrena A.,National Agricultural Research Institute INIA | Rubio L.,National Agricultural Research Institute INIA | Gonzalez-Arcos M.,National Agricultural Research Institute INIA | Maeso D.,Estacion Experimental W. F. Aldunate | And 2 more authors.
Plant Disease | Year: 2016

During 2013, a survey of greenhouse tomato plants with virus-like symptoms was conducted in Salto (Northwestern Uruguay), the main greenhouse tomato-producing area in Uruguay. High populations of the whitefly Bemisia tabaci (Hemiptera:Aleyrodidae) were present, with a number of tomato plants displaying mosaic and slight crumpling in young leaves. Leaf samples were collected from six symptomatic plants from three greenhouses (two plants were sampled per greenhouse) as well as from asymptomatic (putative healthy) plants used as negative controls. Total DNA was isolated from each sample and subjected to PCR using PAL1v1978/PAR1c496 primer set (Rojas et al. 1993), since begomovirus (genus Begomovirus, family Geminiviridae) infection was suspected. Amplification of the expected 1,100-bp fragment was observed in all samples from symptomatic plants whereas no such amplification was observed in samples from asymptomatic plants. The PCR fragments were directly sequenced, and 98 to 99% nucleotide sequence identity was found among the six sequences obtained. BLAST sequence analysis revealed ca. 94% identity to the sequence of the begomovirus Tomato yellow veins streak virus (ToYVSV) reported from Argentina (GenBank Accession No. KJ413253). DNA from one of the samples (SLT_43) was used for rolling-circle amplification (RCA) using ϕ29 DNA polymerase (TempliPhi kit, GE Healthcare, Little Chalfont, UK). Restriction pattern of the RCA product using HpaII suggested a bipartite nature for the genome of this begomovirus yielding four fragments that total approximately 5.1 kbp in size (1.9, 1.8, 0.8, and 0.6 kbp each). Putative DNA-A and DNA-B genome components were then cloned using RCA products digested with PstI and BamHI endonucleases that have single-cleavage sites in each component, respectively. The corresponding inserts for each component were cloned and completely sequenced (KR024026 and KR024027, respectively). The cloned components exhibited the typical genome organization of New World bipartite begomoviruses (Brown et al. 2012), with a 160-nt common region exhibiting 98% identity between A and B components and indicating that both represent a cognate pair. The DNA-A sequence was aligned with available begomovirus sequences from the GenBank database using MUSCLE, and pairwise identity scores were calculated using SDT (Sequence Demarcation Tool) (Muhire et al. 2014). The analysis showed that the DNA-A sequence displayed maximum (96%) nucleotide sequence identity with that of a ToYVSV tomato isolate from Argentina (KJ413253). Similar analysis for DNA-B also indicated maximum (89%) sequence identity with that of several isolates of ToYVSV available in databases. Therefore, based on the latest species demarcation criteria for begomoviruses (Brown et al. 2015), the characterized virus is an isolate of ToYVSV. To our knowledge, this is the first report of this virus infecting tomato in Uruguay. © 2016 The American Phytopathological Society.


Couto D.,Norwich Research Park | Stransfeld L.,Norwich Research Park | Arruabarrena A.,National Agricultural Research Institute INIA | Zipfel C.,Norwich Research Park | And 2 more authors.
BMC Research Notes | Year: 2015

Background: Standard molecular biological methods involve the analysis of gene expression in living organisms under diverse environmental and developmental conditions. One of the most direct approaches to quantify gene expression is the isolation of RNA. Most techniques used to quantify gene expression require the isolation of RNA, usually from a large number of samples. While most published protocols, including those for commercial reagents, are either labour intensive, use hazardous chemicals and/or are costly, a previously published protocol for RNA isolation in Arabidopsis thaliana yields high amounts of good quality RNA in a simple, safe and inexpensive manner. Findings: We have tested this protocol in tomato and wheat leaves, as well as in Arabidopsis leaves, and compared the resulting RNA to that obtained using a commercial phenol-based reagent. Our results demonstrate that this protocol is applicable to other plant species, including monocots, and offers yield and purity at least comparable to those provided by commercial phenol-based reagents. Conclusions: Here, we show that this previously published RNA isolation protocol can be easily extended to other plant species without further modification. Due to its simplicity and the use of inexpensive reagents, this protocol is accessible and affordable and can be easily implemented to work on different plant species in laboratories worldwide. © 2015 Couto et al.; licensee BioMed Central.


Carrasco-Letelier L.,National Agricultural Research Institute INIA | Mendoza-Spina Y.,National Agricultural Research Institute | Branchiccela M.B.,Institute Investigaciones Biologicas Clemente Estable
Chemosphere | Year: 2012

Glyphosate-resistant soybean cultivation is expanding rapidly in Uruguay, with its land area having increased by 95 times during the past 10years. Because of the region's Neotropical conditions, insecticide use is required to ensure adequate soybean productivity. However, in areas shared by soybean crops and beekeepers - such as the southwestern zone of Uruguay (SWZU) - the use of insecticides can increase the risks of honeybee death and honey contamination. Uruguayan commercial and legal guidelines set out practices and field doses designed to prevent acute intoxication with insecticides. However, honeybees in the SWZU are predominantly a polyhybrid subspecies different from that used to set international reference values, and hence they may have a different acute toxicity response, thus rendering such precautions ineffective. The aim of this work was to assess the acute toxicity response of polyhybrid honeybees in the SWZU to cypermethrin (commercial formulation: Cipermetrina 25 Agrin®), chlorpyrifos (commercial formulation: Lorsban 48E®), and endosulfan (commercial formulation: Thionex 35®). Acute toxicity bioassays were conducted to determine the median lethal dose (LD50) of each insecticide for the honeybees. The results indicate that, compared with EU reference values, SWZU honeybees have a higher toxicological sensitivity to chlorpyrifos and endosulfan, and a lower toxicological sensitivity to cypermethrin, based on the commercial formulations tested. However, when these results were adjusted according to their field dose equivalents, only chlorpyrifos emerged as a potential problem for beekeeping, as the maximum recommended field dose of Lorsban 48E® for soybean crops in Uruguay is 23 times the corresponding LD50 for honeybees in the SWZU. © 2012 Elsevier Ltd.


Rivas-Rivera N.,University of the Republic of Uruguay | Eguren G.,University of the Republic of Uruguay | Carrasco-Letelier L.,National Agricultural Research Institute INIA | Munkittrick K.R.,University of New Brunswick
Ecotoxicology | Year: 2014

Sediment constitutes an important sink of endocrine disruptor compounds; however, the potential of sediments to act as a source of endocrine disruptors should be more extensively investigated. The main objective of this study was to determine whether exposure of immature common carp to Uruguay River sediments undergo physiological and endocrine alterations. The lower Uruguay River watershed supports intensive agricultural and forest production, receives municipal sewage discharge and industrial effluent, and a new large pulp mill was constructed in 2006. A 30-day semi-static assay was performed using sediments from four sites along the Uruguay River and compared with an unexposed group in dechlorinated water as a negative control. We focused on two upstream and two downstream sites of a new elemental chlorine free pulp mill. The results showed that plasma vitellogenin levels increased in fish along the river and significant differences were found between the exposed and unexposed groups. Condition factor and gonadosomatic index were not different; however, a significant difference in hepatosomatic index was observed in fish exposed to sediment from an industrial site. A significant reduction in primary spermatocyte accumulation was observed in the exposed group compared with that in the control group, and some individuals exposed to sediments from industrial sites presented with testis-ova. Our results suggest that Uruguay River sediments act as an important source of estrogenic compounds that could be responsible for the alterations observed. Future studies are needed to identify the causal agents and determine exposure routes. © 2014 Springer Science+Business Media.


PubMed | National Agricultural Research Institute INIA and Norwich Research Park
Type: | Journal: BMC research notes | Year: 2015

Standard molecular biological methods involve the analysis of gene expression in living organisms under diverse environmental and developmental conditions. One of the most direct approaches to quantify gene expression is the isolation of RNA. Most techniques used to quantify gene expression require the isolation of RNA, usually from a large number of samples. While most published protocols, including those for commercial reagents, are either labour intensive, use hazardous chemicals and/or are costly, a previously published protocol for RNA isolation in Arabidopsis thaliana yields high amounts of good quality RNA in a simple, safe and inexpensive manner.We have tested this protocol in tomato and wheat leaves, as well as in Arabidopsis leaves, and compared the resulting RNA to that obtained using a commercial phenol-based reagent. Our results demonstrate that this protocol is applicable to other plant species, including monocots, and offers yield and purity at least comparable to those provided by commercial phenol-based reagents.Here, we show that this previously published RNA isolation protocol can be easily extended to other plant species without further modification. Due to its simplicity and the use of inexpensive reagents, this protocol is accessible and affordable and can be easily implemented to work on different plant species in laboratories worldwide.


Stewart S.,National Agricultural Research Institute INIA | Abeysekara N.,Iowa State University | Robertson A.E.,Iowa State University
Plant Disease | Year: 2014

Changes in pathotype structure of Phytophthora sojae populations have been attributed to deployment of race-specific resistant Rps genes in soybean that have been incorporated into commercial cultivars to reduce losses due to Phytophthora root and stem rot. To test this hypothesis, a cultivar rotation study was established from 2007 through 2010 in microplots at a site in Iowa with no history of soybean cultivation. All microplots were inoculated with P. sojae isolate PR1, race 1 (vir 7) prior to planting in year 1. Six rotations were tested: (i) continuous planting of a P. sojae-susceptible cultivar, (ii) continuous planting of a cultivar with high partial resistance to the pathogen, (iii) continuous planting of a cultivar with the Rps 1k gene, (iv) annual rotation of a susceptible with a resistant cultivar, (v) annual rotation of a partially resistant cultivar with a cultivar with the Rps 1k gene, and (vi) 4-year rotation of cultivars with Rps 1k, 1c, 3a, and 1k genes in year one, two, three, and four, respectively. The diversity of 121 isolates of P. sojae that were recovered by baiting from soil samples collected from the experiment were assessed using pathotyping and eight microsatellite markers, and compared with PR1. Changes in pathotype and multilocus genotypes (MLGs) were recorded at the second sampling date, indicating that P. sojae has the ability to evolve quickly. In total, 14 pathotypes and 21 MLGs were recovered over the 4-year experiment, and only 49 and 22% of the isolates had the same pathotype and MLG, respectively, as PR1. The number of isolates of P. sojae recovered varied among rotations, with more isolates recovered from rotations that included a cultivar with partial resistance. Gain of virulence was detected on Rps 1a, 1b, 1c, 1d, and 3a and was not dependent on rotation. Using simple-sequence repeat analysis, 10 alleles that were different from those of PR1 were detected throughout the 4-year period. Cultivar rotation affected the genetic structure of the P. sojae population. Recovery of isolates with different MLGs, genotypic diversity (G = 4.7), and gene diversity (UHe = 0.45) were greater under continuous rotation with partial resistance. Phytophthora root and stem rot causes economic losses in the north-central region of the United States annually. An improved understanding of the effect of Rps gene deployment on P. sojae diversity would lead to improved management practices and reduced losses. © 2014 The American Phytopathological Society.


Ibanez F.,National Agricultural Research Institute INIA | Zoppolo R.,National Agricultural Research Institute INIA
Allelopathy Journal | Year: 2011

In plantations of B. dracunculifolia, a noticeable decrease in diversity and development of monocots species was observed. Hence, phytotoxic studies were done to determine the in vitro toxicity of essential oils of B. dracunculifolia on monocot (Lolium multiflorum) and dicot (Lactuca sativa). The essential oil at concentration of 1000 mg/L drastically decreased the hypocotyl growth of L. multiflorum (90%) and L. sativa (60%). The analysis of essential oil by gas chromatography showed a high percentage of compounds with phytotoxic effects, such as limonene, α and β-pinene. The field experiments showed that the inhibition under B. dracunculifolia was not due to the effect of shading nor to nutrient competition. The reduction in germination of L. multiflorum under B. dracunculifolia was > 90%. In the soil under the plantation, a high concentration of phenolics was measured (15mg/100g of soil). These results indicate that allelopathic activity of B. dracunculifolia may be explained through a combination of phytotoxic effects (emission of volatile compounds and the accumulation of phenolics in the soil profile).


PubMed | National Agricultural Research Institute INIA
Type: Journal Article | Journal: Chemosphere | Year: 2012

Glyphosate-resistant soybean cultivation is expanding rapidly in Uruguay, with its land area having increased by 95 times during the past 10 years. Because of the regions Neotropical conditions, insecticide use is required to ensure adequate soybean productivity. However, in areas shared by soybean crops and beekeepers - such as the southwestern zone of Uruguay (SWZU) - the use of insecticides can increase the risks of honeybee death and honey contamination. Uruguayan commercial and legal guidelines set out practices and field doses designed to prevent acute intoxication with insecticides. However, honeybees in the SWZU are predominantly a polyhybrid subspecies different from that used to set international reference values, and hence they may have a different acute toxicity response, thus rendering such precautions ineffective. The aim of this work was to assess the acute toxicity response of polyhybrid honeybees in the SWZU to cypermethrin (commercial formulation: Cipermetrina 25 Agrin), chlorpyrifos (commercial formulation: Lorsban 48E), and endosulfan (commercial formulation: Thionex 35). Acute toxicity bioassays were conducted to determine the median lethal dose (LD(50)) of each insecticide for the honeybees. The results indicate that, compared with EU reference values, SWZU honeybees have a higher toxicological sensitivity to chlorpyrifos and endosulfan, and a lower toxicological sensitivity to cypermethrin, based on the commercial formulations tested. However, when these results were adjusted according to their field dose equivalents, only chlorpyrifos emerged as a potential problem for beekeeping, as the maximum recommended field dose of Lorsban 48E for soybean crops in Uruguay is 23 times the corresponding LD(50) for honeybees in the SWZU.

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