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SummaryStandard models of plant speciation assume strictly dichotomous genealogies in which a species, the ancestor, is replaced by two offspring species. The reality in wind-pollinated trees with long evolutionary histories is more complex: species evolve from other species through isolation when genetic drift exceeds gene flow; lineage mixing can give rise to new species (hybrid taxa such as nothospecies and allopolyploids). The multi-copy, potentially multi-locus 5S rDNA is one of few gene regions conserving signal from dichotomous and reticulate evolutionary processes down to the level of intra-genomic recombination. Therefore, it can provide unique insights into the dynamic speciation processes of lineages that diversified tens of millions of years ago. Here, we provide the first high-throughput sequencing (HTS) of the 5S intergenic spacers (5S-IGS) for a lineage of wind-pollinated subtropical to temperate trees, theFagus crenata – F. sylvaticas.l. lineage, and its distant relativeF. japonica.The observed 4,963 unique 5S-IGS variants reflect a complex history of hybrid origins, lineage sorting, mixing via secondary gene flow, and intra-genomic competition between two or more paralogous-homoeologous 5S rDNA lineages. We show that modern species are genetic mosaics and represent a striking case of ongoing reticulate evolution during the past 55 million years.Significance statementThe evolution of extra-tropical wind-pollinated tree genera involves dynamic speciation processes. High-throughput sequencing of the multi-copy, potentially multi-locus 5S rDNA reveals a complex history of hybrid origins, lineage sorting and mixing, and intra-genomic competition between paralogous-homeologous loci in the core group of Eurasian beech trees (genusFagus) and their distant relative,F. japonica. The modern species are genetic mosaics and represent a striking case of at least 55 million years of ongoing reticulate evolution.

The aim of this study is to better understand the phylogeography and evolution of the Lebanese oaks and to assess the convergence between their morpho-ecological and molecular characteristics. We i...

Gastridium P. Beauv. is a Mediterranean-Paleotropical member of the Poaceae family, whose species number and diversity are still imperfectly known. We examined patterns of DNA diversity at two plas...

This study aimed to disentangle the structure, composition, and co-occurrence relation-ships of the banana (cv. Dwarf Cavendish) root endophytome comparing two phenological plant stages: mother plants and suckers. Moreover, a collection of culturable root endophytes (>1000) was also generated from Canary Islands. In vitro antagonism assays against Fusarium oxysporum f.sp. cubense (Foc) races STR4 and TR4 enabled the identification and characterization of potential biocontrol agents (BCA). Eventually, three of them were selected and evaluated against Fusarium wilt of banana (FWB) together with the well-known BCA Pseudomonas simiae PICF7 under controlled conditions. Culturable and non-culturable (high-throughput sequencing) approaches provided concordant information and showed low microbial diversity within the banana root en-dosphere. Pseudomonas appeared as the dominant genus and seemed to play an important role in the banana root endophytic microbiome according to co-occurrence networks. Fungal communities were dominated by the genera Ophioceras, Cyphellophora, Plecosphaerella, and Fusarium. Overall, significant differences were found between mother plants and suckers, suggesting that the phe-nological stage determines the recruitment and organization of the endophytic microbiome. While selected native banana endophytes showed clear antagonism against Foc strains, their biocontrol performance against FWB did not improve the outcome observed for a non-indigenous reference BCA (strain PICF7)., This study was supported by Project ‘MUSA’ (Microbial Uptakes for Sustainable management of major bananA pests and diseases), funded by the EU’s Horizon 2020 Research and Innovation Program (Grant Agreement Identifier: 727624).

[Background] Verticillium wilt of olive (VWO) is caused by the soilborne fungal pathogen Verticillium dahliae. One of the best VWO management measures is the use of tolerant/resistant olive cultivars. Knowledge on the olive-associated microbiome and its potential relationship with tolerance to biotic constraints is almost null. The aims of this work are (1) to describe the structure, functionality, and co-occurrence interactions of the belowground (root endosphere and rhizosphere) microbial communities of two olive cultivars qualified as tolerant (Frantoio) and susceptible (Picual) to VWO, and (2) to assess whether these communities contribute to their differential disease susceptibility level., [Results] Minor differences in alpha and beta diversities of root-associated microbiota were detected between olive cultivars regardless of whether they were inoculated or not with the defoliating pathotype of V. dahliae. Nevertheless, significant differences were found in taxonomic composition of non-inoculated plants’ communities, “Frantoio” showing a higher abundance of beneficial genera in contrast to “Picual” that exhibited major abundance of potential deleterious genera. Upon inoculation with V. dahliae, significant changes at taxonomic level were found mostly in Picual plants. Relevant topological alterations were observed in microbial communities’ co-occurrence interactions after inoculation, both at structural and functional level, and in the positive/negative edges ratio. In the root endosphere, Frantoio communities switched to highly connected and low modularized networks, while Picual communities showed a sharply different behavior. In the rhizosphere, V. dahliae only irrupted in the microbial networks of Picual plants., [Conclusions] The belowground microbial communities of the two olive cultivars are very similar and pathogen introduction did not provoke significant alterations in their structure and functionality. However, notable differences were found in their networks in response to the inoculation. This phenomenon was more evident in the root endosphere communities. Thus, a correlation between modifications in the microbial networks of this microhabitat and susceptibility/tolerance to a soilborne pathogen was found. Moreover, V. dahliae irruption in the Picual microbial networks suggests a stronger impact on the belowground microbial communities of this cultivar upon inoculation. Our results suggest that changes in the co-occurrence interactions may explain, at least partially, the differential VWO susceptibility of the tested olive cultivars., Supported by grant AGL2016-75729-C2-1-R from the Spanish Ministerio de Economía, Industria y Competitividad/Agencia Estatal de Investigación, and co-financed by the European Regional Development Fund (ERDF).

Agriculture and urban development have been the main drivers of loss and fragmentation of grasslands worldwide. The Argentine Pampas has been highly transformed by human activities. However how birds respond to the size of grassland patches and landscape matrices is unknown. We studied the effects of these on the abundance and richness of both specialist and opportunist grassland birds. In addition, we evaluated the patch size effect in contrast with unfragmented grasslands. We surveyed birds in small and large patches of Cortaderia selloana grasslands embedded within agricultural, planted forest, dune and urban landscape matrices and, specifically in spring, in unfragmented grasslands. The abundance and richness of specialist grassland birds in small patches were lower than in large patches, but richness depended on the type of matrix and was lowest in patches surrounded by a forest matrix. Extensive grasslands are a key habitat for grassland specialists during the breeding season. In contrast, the abundance and richness of opportunist grassland birds were higher in patches than in unfragmented grasslands, and showed a negative effect of dune matrix in winter. Our results enable prediction of how bird species with different habitat requirements may vary in abundance and richness depending on the size of grassland patches and the type of land use following grassland replacement. El desarrollo agrícola y urbano han sido las principales causas de pérdida y fragmentación de los pastizales en todo el mundo. La región Pampeana de Argentina ha sido enormemente transformada por las actividades humanas; sin embargo se desconoce cómo las aves responden al tamaño de los parches de pastizal remanentes y a las matrices de paisaje. Estudiamos ambos efectos sobre la abundancia y riqueza de aves especialistas y oportunistas de pastizal. Además, evaluamos el efecto del tamaño del parche contrastándolo con pastizales sin fragmentar. Muestreamos aves en parches pequeños y grandes de Cortaderia selloana inmersos en matrices agrícolas, forestales, de dunas y urbanas, y específicamente en primavera, en pastizales sin fragmentar. La abundancia y riqueza de aves especialistas en parches pequeños fueron menores que en parches grandes, pero la riqueza dependió del tipo de matriz, y la menor ocurrió en parches rodeados de matriz forestal. Los extensos pastizales son un hábitat clave para las aves especialistas durante la estación reproductiva. Por el contrario, la abundancia y riqueza de aves oportunistas fueron mayores en parches que en pastizales sin fragmentar, y mostraron un efecto negativo de la matriz de dunas durante el invierno. registramos un efecto del tamaño del parche sobre aves especialistas; sin embargo, este efecto estuvo modulado por el uso del suelo que domina el paisaje alrededor de los parches. Nuestros resultados permiten predecir cómo las aves, con diferentes requerimientos de hábitat, pueden variar en abundancia y riqueza dependiendo del tamaño del parche de pastizal y del uso de la tierra derivado de su reemplazo. Fil: Pretelli, Matías Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Isacch, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Cardoni, Daniel Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina

Between 2013-2015, we conducted a study on the breeding biology of the Wren-like Rushbird (Phleocryptes melanops) in wetlands of the Pampas region, Argentina. The search for rushbird nests was conducted by several researchers walking into the wetlands, in three ways: by observing adults carrying material, through singing adults next to the nest or from the active search for nests in the vegetation. Nests were visited at intervals of 3-5 days. Of the 245 nests that we encountered (153 nests in 2013-2014 and 92 nests in 2014-2015), we found eggs in 97. The egg-laying period lasted almost 3 months (late Sept-late Dec). The mean clutch size was 2.7 ± 0.5 eggs (mean ± SD) (range = 1-3; n = 63), and the total nesting period was 34 ± 2.5 days (mean ± SD), with incubation and chick-rearing periods of 18 ± 1.63 days and 16 ± 0.50 days, respectively. From the total number of nests found, 43% of them were abandoned during building, 38% of nests were depredated, 14% of nests (n = 33) were successful, while 5% of nests were destroyed. Nest predation was higher during the incubation (80%) than during chick-rearing period (20%). The nesting success was on average of 23% (25% for the 2013-14 and 21% for the 2014-2015) for the entire nesting period. We also observed relatively higher plasticity than previously reported in relationship to vegetation used to fix nests (80% rushes, 20% other plants). Finally, we observed an abrupt termination of the reproductive period after desiccation of the wetlands; 90% of active nests failed (75% depredated and 25% abandoned) when the wetland was dried. Fil: Chiaradia, Nicolas Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Cardoni, Daniel Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Pretelli, Matías Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Isacch, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina

Incomplete taxonomic knowledge may seriously hamper biodiversity conservation efforts that are crucial in a context of global change.GastridiumP.Beauv. is a Mediterranean-Paleotropical member of the Poaceae family, inhabiting ephemeral grass habitats, whose species number and diversity are still imperfectly known. In order to progress towards a comprehensive taxonomic treatment of this genus, we examined patterns of DNA diversity in the four taxa (Gastridium lainzii,G. phleoides,G. scabrum, andG. ventricosum) that have been recently advanced by different authors, based on new morpho-ecological descriptors. We explored nucleotide sequence variation at two plastid (trnH-psbA, trnL-F) and one nuclear (ITS) DNA markers in 44 total individuals. Diversity data were treated with multiple statistical and phylogenetic tools, and integrated with available GenBank sequences ofGastridiumand other closely related genera. Despite the limited variability detected, evidence of within-taxon genetic cohesion and estimates of molecular divergence comparable with those of species in the same subtribal lineage (Agrostidinae) were recovered. The identified plastid genealogies appeared congruent with a subdivision of the genus into (at least) three distinct entities, and coherent with collected morphological descriptors. Phylogenetic reconstructions with ITS were less corresponding to taxa identities, likely due to reticulation and polyploidization. Once placed in a broader taxonomic context, the investigated dataset produced plastid and nuclear tree topologies consistent with previous assessments, highlighting the overall little resolution of species and genera within Agrostidinae. In the plastid tree, a sister relationship betweenGastridiumandTriplachnewas weakly supported. In the ITS tree, relationships among these genera were unresolved. The hypothesis of closely related but separately evolving lineages withinGastridiumis discussed, suggesting a re-evaluation of its current assessment in taxonomic authorities to enhance our knowledge of the grass family, and assist future biodiversity surveys of key Mediterranean grassland ecosystems.

Diverse factors may limit nest-site selection; however, nest predation has long been considered a main factor driving nest-site selection in birds. Marsh-nesting birds face a particularly important trade-off between habitat features that impede their vision of the approach of potential predators and nest concealment from predators. Here, we analyse how different ecological variables influence nest-site selection in the habitat-specialist Wren-like Rushbird (Phleocryptes melanops) breeding in three different wetlands of the Pampas region of Argentina over 3 years. Rushbirds selected nest sites with higher heights and denser vegetation, compared to random sites. Nest height was positively associated with vegetation height and lower visibility from the point of view of aerial predators, and the nest entrance was oriented towards the region with greatest visibility of the nest from the point of view of Rushbirds. Rushbirds seem to flexibly adjust their nest location in response to high predation risk, building their nests under structures built by other birds when breeding colonies of herons and gulls (potential nest predators) are present. Initial nest-site selection is performed by males, which then attempt to attract females to a rudimentary nest. Females sometimes reject such sites and the male then selects a new nesting location. Our study highlights the potential for flexible sex differences in nest-site selection. Fil: Chiaradia, Nicolas Mariano. Universidad Nacional de Mar del Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Isacch, Juan Pablo. Universidad Nacional de Mar del Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Pretelli, Matías Guillermo. Universidad Nacional de Mar del Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Cardoni, Daniel Augusto. Universidad Nacional de Mar del Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina

Verticillium wilt of olive (VWO), caused by the soil-borne pathogen Verticillium dahliae, is considered one of the most important diseases affecting this tree crop. One of the best VWO management measures is the use of tolerant cultivars. Remarkably, no information is available about olive functional traits and their potential relationship with tolerance to V. dahliae. Twenty-five selected functional traits (for leaf, stem, root and whole plant) were evaluated in six olive varieties differing in their VWO tolerance level to identify possible links between this phenotype and functional traits’ variation. High intervarietal diversity was found among cultivars and several functional traits were related with VWO tolerance. Tolerant varieties showed higher leaf area, dry matter content (leaf, stem and plant) and mass fraction for stems, but lower for leaves. Significant differences were also detected for root functional traits, tolerant cultivars displaying larger fine root diameter and lignin content but smaller specific length and area of thick and fine roots. Correlations were found among functional traits both within varieties and between levels of tolerance/susceptibility to VWO. Associations were observed between biomass allocation, dry matter content and VWO tolerance. The most relevant difference between tolerant and susceptible cultivars was related to root system architecture., This work was supported by the Spanish Ministerio de Ciencia e Innovación (grant ECOMEDIT CGL2014-53236-R), Ministerio de Economía, Industria y Competitividad (grant AGL2016-75729-C2-1-R), Ministerio de Ciencia e Innovación/Agencia Estatal de Investigación (grant PID2019-106283RB-I00), and by the Junta de Andalucía (Spain) grants ‘For-Change’ (UCOFEDER 18 REF 27943 MOD B) and P18-RT-3455, all of them cofunded with European FEDER funds.

The application of anaerobically digested cattle manure on agricultural land for both improving its quality and recycling a farm waste is an increasingly frequent practice in line with the circular economy. However, knowledge on the potential risk of spreading antibiotic resistance through this specific practice is quite scarce. The antibiotic sulfamethoxazole (SMX) is one of the most heavily prescribed in veterinary medicine. In this study, SMX dissipation and the possible effects on natural microorganisms were investigated in a soil amended with an anaerobically digested cattle manure produced from a biogas plant inside a livestock farm. Microcosm experiments were performed using amended soil treated with SMX (20 mg/kg soil). During the experimental time (61 days), soil samples were analysed for SMX and N4-acetylsulfamethoxazole, microbial abundance, activity and structure. Furthermore, the prevalence of the intI1 gene was also determined. The overall results showed that, although there was an initial negative effect on microbial abundance, SMX halved in about 7 days in the digestate-amended soil. The intI1 gene found in both the digestate and amended soil suggested that the use of anaerobically digested cattle manure as fertilizer can be a source of antibiotic resistant bacteria (ARBs) and genes (ARGs) in agroecosystems.

Predation risk on birds is often an important source of natural selection that shapes parental care and may promote behavioral changes. Parents can often estimate certain risks and adjust their behavior to reduce the likelihood of nest predation. The fragmentation of habitats is one of the main consequences of loss of habitats, and in general, for birds breeding in smaller patches, their daily nest-survival rate is lower due to increased nest predation. Since nest survival is an estimate of predation risk in the environment, we evaluated the daily survival rate (DSR) for nests of spectacled tyrants (Hymenops perspicillatus) and parental care behavior on fragmented and unfragmented grasslands. We conducted nest searching and monitoring during the 2012-2013 breeding season in small patches and in a continuous patch of grassland. In addition, parental activity was recorded using video monitoring. We found a lower DSR for the spectacled tyrant in fragmented grasslands, associated with increased nest predation risk; females showed a variation in parental care. This variation was evidenced by larger incubation bouts and lower visitation rate during the incubation period, and by a lower food delivery rate to nestlings, compensated by larger prey sizes. The results show that fragmentation not only reduces the fitness of individuals and impacts adversely on population, but individuals are also subjected to a strong selection pressure, and their reproductive success may depend to some extent on the ability of parents to estimate at least certain predation risk and adjust their behavior in this regard. Fil: Pretelli, Matías Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Isacch, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Cardoni, Daniel Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina

Accurate identification of aquatic species is fundamental to freshwater research. In this paper, we targeted Ephemeroptera, a key taxonomic insect group for biomonitoring of water bodies and present an overview on the efficacy of the DNA barcoding approach to document species identity in the Mediterranean region. We sequenced the mitochondrial cytochrome c oxidase (COI) in 39 nominal species. Sample discrimination and species identification were investigated by evaluating haplotype identity and similarity, intra-/interspecific genetic distances, optimal identification of barcoding gap thresholds, estimates of species monophyly and comparative species matches on available reference libraries. The resolving power of the obtained data was discussed in the light of statistical tools such as Spider R-package and Poisson Tree Processes. High levels of species identification were achieved with all the used methodologies, and the occurrence of cryptic species was suggested. We conclude that DNA barcoding is a powerful tool for taxonomic research in Mediterranean mayflies, with great promise to ameliorate biodiversity inventories of freshwater ecosystems and to provide the necessary accuracy for water quality assessment programs. Our results further indicated we need to upgrade the current regional mayfly diversity knowledge. The development of a Mediterranean reference library could integrate this new information system.

The loss of grasslands in southeastern South America has negatively affected grassland birds, leading to marked declines in their populations. However, the extent to which habitat transformation impacts on their reproductive performance, and whether the magnitude of these effects may be modulated by landscape matrices, is unknown. We assessed the effect of fragmentation on grassland bird reproduction by comparing the combined influence of fragmentation and landscape matrix on nesting success, brood parasitism and productivity of the Spectacled Tyrant Hymenops perspicillatus and the Brown-and-yellow Marshbird Pseudoleistes virescens. Surveys were undertaken in small grassland patches embedded within different landscape matrices (urban and agro-ecosystem) and in a large patch within a reserve. Reproductive performance was adversely affected by fragmentation. However, these effects were conditioned by matrix type, and the response was not the same for the two species. For Brown-and-yellow Marshbird, fragmentation resulted in higher rates of brood parasitism and lower productivity regardless of the matrix type, whereas for Spectacled Tyrant, we found a negative effect only in an agricultural matrix. The lack of extensive grasslands makes small patches important; however, knowing the effects of different matrix types is critical to predicting the conservation value of grassland patches, and the response of different species is not uniform. Fil: Pretelli, Matías Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Isacch, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Cardoni, Daniel Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina

Saltmarshes of Spartina densiflora in south-eastern South America have been modified by anthropogenic activities, mainly production of livestock. We examined the effect of the intensity of cattle production on the structure of saltmarsh vegetation and the effect of these changes to vegetation on the richness, composition and size of the avian populations and the abundance of nests. The levels of cattle production were based on the combined intensity of prescribed burning and cattle grazing, classed as: (1) High grazing - High burning (HH), (2) Low grazing - Low burning (LL) and (3) No grazing - No burning (NN). Cattle production altered the vegetation structure of saltmarshes and indirectly modified the richness, composition and size of their avian populations and the abundance of nests. Saltmarshes with either LL or NN production levels were inhabited by tall grassland specialists and generalists and by species specialised to live in a mosaic of short and tall grassland patches. Conversely, saltmarshes with HH production levels were inhabited by short-grassland specialists. That avian species diversity does not differ between S. densiflora saltmarsh subject to low or no human impacts has several potential interpretations, which are discussed. These findings have implications for management of grasslands to maintain avian diversity. Journal compilation Fil: Cardoni, Daniel Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Isacch, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Iribarne, Oscar Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina

According to the current criteria, the diagnosis of early allograft dysfunction usually cannot be established before the end of the first week after liver transplantation. Thus, early predictive tests for detecting allograft dysfunction are still warranted to prevent allograft failure. This study was undertaken to assess the role of low serum factor V activity as an early prognostic factor (postoperative day 2) after liver transplantation.A retrospective review of all consecutive adult patients who underwent first orthotopic whole-graft liver transplant at our institution between March 2002 and June 2011 was undertaken. Primary endpoint was graft failure within 90 days after transplantation.Of all 105 patients analyzed in this study, 39 (37.1 %) were female and 66 (62.9 %) were male. Mean age was 52.7 ± 11.7 years, and median follow-up period was 2474 ± 164 days. There were overall 33 (31.4 %) deaths, 13 of those occurring on the first 90 post-transplant days. Multivariate analysis demonstrated that serum factor V lower than 41.5 % and female gender had a negative impact not only on allograft failure/death within 90 days after transplantation (RR = 5.30, CI = 1.40-20.2, p = 0.015 and RR = 5.23, CI = 1.53-21.33, p = 0.008) but also on overall mortality. For prediction of allograft failure/death occurring during the first 3 months, serum factor V level of 41.5 % or lower exhibited a specificity of 87.9 %, a sensitivity of 42.9 %, an accuracy of 81.9 %, a positive predictive value of 35.3 %, and a negative predictive value of 90.9 %.Assessment of serum factor V levels on postoperative day 2 might be a promising prognostic tool for early prediction of inferior outcomes after liver transplantation.

Nucleotide sequences from the plastome are currently the main source for assessing taxonomic and phylogenetic relationships in flowering plants and their historical biogeography at all hierarchical levels. One major exception is the large and economically important genusQuercus(oaks). Whereas differentiation patterns of the nuclear genome are in agreement with morphology and the fossil record, diversity patterns in the plastome are at odds with established taxonomic and phylogenetic relationships. However, the extent and evolutionary implications of this incongruence has yet to be fully uncovered. The DNA sequence divergence of four Euro-Mediterranean Group Ilex oak species (Quercus ilexL.,Q. cocciferaL.,Q. aucheriJaub. & Spach.,Q. alnifoliaPoech.) was explored at three chloroplast markers (rbcL, trnK/matK, trnH-psbA). Phylogenetic relationships were reconstructed including worldwide members of additional 55 species representing allQuercussubgeneric groups. Family and order sequence data were harvested from gene banks to better frame the observed divergence in larger taxonomic contexts. We found a strong geographic sorting in the focal group and the genus in general that is entirely decoupled from species boundaries. High plastid divergence in members ofQuercusGroup Ilex, including haplotypes shared with related, but long isolated oak lineages, point towards multiple geographic origins of this group of oaks. The results suggest that incomplete lineage sorting and repeated phases of asymmetrical introgression among ancestral lineages of Group Ilex and two other main Groups of Eurasian oaks (Cyclobalanopsis and Cerris) caused this complex pattern. Comparison with the current phylogenetic synthesis also suggests an initial high- versus mid-latitude biogeographic split withinQuercus. High plastome plasticity of Group Ilex reflects geographic area disruptions, possibly linked with high tectonic activity of past and modern distribution ranges, that did not leave imprints in the nuclear genome of modern species and infrageneric lineages.

Bay-capped Wren-Spinetail Spartonoica maluroides (Furnariidae) nests very close to the ground in dense clumps of grasses or sedges and builds both open and enclosed nest. We describe a proportion and characteristics of both types of nests and evaluate some causes that could force the construction of one nest architecture or another. In order to study the causes of this variation, we assessed the variability of nest types (architecture and size) and vegetation structure, and examined their consequences for breeding parameters in a population of Bay-capped Wren-Spinetail breeding in Spartina densiflora saltmarshes located at a coastal lagoon on Atlantic coast in east Argentina. We found that Bay-capped Wren-Spinetail builds nests enclosed and open with similar frequency within the same population and vegetation type. All nests were built with stems and grass leaves of S. densiflora. We found that open nests were built in sites with denser vegetation than enclosed nests. Nesting success and nest survival were not affected by nest architecture and size. The ability to adjust nest structure according to the vegetation density may be a strategy aimed at increasing nest concealment to reduce the temperature inside the nest or to avoid nest depredation. A flexible nest architecture strategy in Bay-capped Wren-spinetail is a possible adaptation to living in simple, yet structurally variable environments such as saltmarshes. Fil: Cardoni, Daniel Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Isacch, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Baladron Felix, Alejandro Victor. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina

We studied the diet of nestling Spectacled Tyrants (Hymenops perspicillatus) in the southeast Pampas region, Argentina. From November 2012 to January 2013, we used video footage to determine prey items that the parents fed to their chicks. We obtained 54 hrs of recording time to survey 18 nests. We identified a total of 125 prey items, representing 33 different taxa, grouped into four classes: Insecta, Arachnida, Chilopoda, and Amphibia. Insects accounted for 94% of total prey. The most frequent prey items were orthopterans (Caelifera), unidentified lepidopteran larvae, and odonats (Zygoptera). Lepidopterans were numerically the most important prey item in November, odonats in December, and orthopterans in January. Diet of the nestling birds was generalist in terms of the consumption of insects, and the changes in prey consumption during the course of the breeding season support an opportunistic feeding behavior by Spectacled Tyrants. Fil: Pretelli, Matías Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Cardoni, Daniel Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Isacch, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina

We studied variation in bird assemblages with plant associations for three different coastal marshes from Southeastern South America (SESA) and assessed how marsh bird assemblages related to nearby upland bird assemblages. We surveyed bird species and plant structure along the tidal gradient of each locality from the low tide level to the upper habitats bordering coastal marshes. Twenty species frequently used coastal marshes, including relatively few migratory species. We found that birds occurring in SESA coastal marshes do not have distributions constrained to coastal marshes. Nonetheless, four bird assemblages were recognized in association with vegetation types and/or sites. Among the recorded coastal marsh species, the bay-capped wren-spinetail (Spartonoica maluroides) is both the most frequent and the most habitat constrained. Bird richness increases steadily along the tidal gradient associated with the increase in vegetation structure, suggesting that bird richness is directly explained by vegetation and indirectly by the physical conditions influencing vegetation structure. Results highlight the importance of SESA middle marshes as habitat for conservation of some threatened SESA grassland birds. Fil: Isacch, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Cardoni, Daniel Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Iribarne, Oscar Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina

Biparental care is the dominant (81 %) form of parental care in birds. The degree of sexual selection and the resulting mating system and parental care type have been determined, in many cases, by inference from external characters of sexual dimorphism. The lack of information and the biological attributes of the tyrant flycatchers (e.g., low sexual dimorphism) make it difficult to determinate the parental care roles in most species. Tyrant flycatchers comprise a diverse bird family, but information on the reproductive behavior of many Tyrannid species is lacking. Our aim was to describe external sexual characters (morphology and coloration) and determine the parental care behavior at the nest in the Warbling Doradito (Pseudocolopteryx flaviventris), especially considering the potential role of sexual differences in relation to parental roles at nest. We studied the parental care behavior during two breeding seasons. Parents were captured to take morphological measurements and a blood sample for molecular sex determination. We found that parental care in the Warbling Doradito was biparental biased toward females, where the incubation was performed exclusively by females and the 74 % of activities of nestling brooding and feeding was performed by females. We also found sexual differences, slight in size and plumage, but more conspicuous in bill color. Despite the relatively slight sexual dichromatism of the Warbling Doradito, the parental care behavior in the nest was biased towards females, and the assistance of males to nestlings was highly variable and was not correlated with either clutch size or increasing nestling mass. Therefore, the evidence presented here suggest that Pseudocolopteryx genus may have an unusual social system, otherwise uncommon in tyrannids. Fil: Cardoni, Daniel Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Pretelli, Matías Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Isacch, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Madrid, Enrique Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Baladron Felix, Alejandro Victor. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Chiaradia, Nicolas Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina

Fil: Cardoni, Daniel Augusto. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina

Oaks of Quercus Group Ilex are emblematic components of the Mediterranean landscapes and the full extent of their diversity in a geographic context is still poorly assessed. In order to detail differentiation patterns within Group Ilex and to illuminate causes and circumstances that underlie the distribution of its lineages, we examined plastome differentiation in the four species currently representing this oak group across the Mediterranean Basin (Quercus ilex, Quercus coccifera, Quercus aucheri, Quercus alnifolia). Plastid DNA variation was explored with three markers on 124 individuals and spatial analyses were performed with phylogenetic and landscape genetics approaches. A strong geographic sorting, entirely decoupled from species boundaries, was found. Spatial coincidences with phylogenetic splits highlights the presence of four geographic regions (the Middle East, the Aegean region, the central and the western Mediterranean) characterised by different degrees of isolation. Distinct sub-regional structures were identified. Inter- and intra-lineage divergence patterns appear to reflect the complex orogeny of the Mediterranean region and Pleistocene climatic oscillations, while a few widespread haplotypes might represent the remnants of the ancestral Group Ilex range. Regional differentiation in this group is recognised to be triggered by a number of factors, including disruption of the original range, incomplete lineage sorting, repeated phases of asymmetrical introgression and isolation. The key role played by some Mediterranean regions on the range establishment and dynamics of this oak group is discussed.

Spartonoica maluroides (Furnariidae) es el ave más especializada y amenazada que habita marismas costeras de esparto de Spartina densiflora del sudeste de América del Sur. Estos ambientes están siendo modificados a un ritmo acelerado principalmente por el pastoreo del ganado y la quema de espartillares, determinando la vulnerabilidad que presenta este furnárido. Para evaluar cómo el uso del suelo afecta las poblaciones de S. maluroides, se estudió la abundancia, el hábitat de nidificación y el éxito reproductivo en marismas bajo diferentes intensidades de pastoreo por ganado y quema. Además, se llevaron a cabo experimentas de depredación de nidos para determinar cómo la altura del nido afecta las tasas de depredación y el tipo de depredadores. S. maluroides presentó similar abundancia, éxito reproductivo y sitio de nidificación en marismas con bajo o nulo pastoreo. Por otro lado, la especie no fue registrada en marismas con alta presión de pastoreo y quemas. La altura del nido fue la variable que mejor explicó la tasa de supervivencia diaria. La tasa de depredación y el tipo de depredadores difirieron en relación a la altura del nido sobre el suelo. Nuestros resultados indican que el pastoreo con baja presión sería compatible con la presencia y reproducción de S. maluroides. De esta manera, el manejo ganadero con baja intensidad de uso es una alternativa viable para el uso sustentable de las marismas del sudeste de América del Sur. The Bay-capped Wren-Spinetail (Furnariidae, Spartonoica maluroides) is the most specialized and threatened bird inhabiting Spartina saltmarshes in southeastern South America. These marshes are being modified at an increased rate, primarily by cattle grazing and burning, which make this furnarid highly vulnerable. To assess how land use affects Bay-capped Wren-Spinetail populations, we estimated their density under different intensities of burnings and cattle grazing, studied nesting habitat selection and nesting success, and conducted nest predation experiments to determine predation rates and predator type. We found similar population densities, nesting microhabitats, and nest predation rates in marshes with low grazing intensity and marshes with no grazing. In contrast, the species was absent from marshes with high intensity of grazing and burning. Nest height was the best predictor of daily survival rates. Nest predation experiments demonstrated that nest high influences predation intensity and predator type. Our results suggest that low grazing intensities are compatible with the presence and reproduction of the Bay-capped Wren-Spinetail. Thus, low-impact cattle raching is a viable alternative for the sustainable development of saltmarshes in southeastern South America. Fil: Cardoni, Daniel Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Isacch, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Iribarne, Oscar Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina

We evaluated how grazing affects bird assemblages in short and tall coastal salty grasslands at Samborombón Bay, Argentina. We studied ranches with three patterns of grazing (spatial rotation, continuous, and winter grazing) and a control site, at each censusing birds and surveying vegetation monthly. Results allowed us to compare the various treatments and their effects on endangered birds of short and tall grassland. Three species of special conservation concern showed significant responses to grazing. In short grasslands, the Buffbreasted Sandpiper (Tryngites subruficollis) is supported by continuous grazing. In tall grasslands, the Bay-capped Wren-Spinetail (Spartonoica maluroides) uses only grasslands with extensive cover of tall grass, whereas Hudson’s Canastero (Asthenes hudsoni) uses those with high levels of cattle grazing. The diverse responses of grassland birds to grazing of different types of grassland leads us to propose a system of heterogeneous grazing that optimizes both production and conservation of grassland birds. Se evaluó de qué manera el pastoreo afecta los ensambles de aves que habitan pastizales halófitos cortos y altos de la bahía Samborombón, Argentina. El estudio se realizó en establecimientos rurales donde se desarrollaban tres tipos de sistemas pastoriles (rotación espacial, pastoreo continuo e invernal) y en un sitio de control. En cada sitio y ambiente, mensualmente se realizaron censos de aves y muestreos de la fisonomía de la vegetación. Los resultados permitieron determinar un contraste entre los tipos de manejo ganadero y sus efectos sobre diferentes aves amenazadas de pastizales altos y cortos. Tres especies de particular importancia para la conservación mostraron respuestas significativas a los tratamientos de pastoreo. Tryngites subruficollis utiliza principalmente pastizales cortos con pastoreo continuo, Spartonoica maluroides utiliza pastizales altos con bajos niveles de pastoreo, y Asthenes hudsoni utiliza pastizales altos con pastoreo continuo y rotativo. Las diversas respuestas de las aves de pastizal al pastoreo en los diferentes tipos de pastizales nos llevan a proponer un sistema de pastoreo heterogéneo que optimice al mismo tiempo la producción y la conservación de las aves de pastizal. Fil: Isacch, Juan Pablo. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Biología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata; Argentina Fil: Cardoni, Daniel Augusto. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Biología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata; Argentina

We studied breeding biology, parental roles, and social mating system of the Bay-capped Wren-Spinetail (Spartonoica maluroides), a habitat specialist furnariid, in the Argentinean pampas. We found 42 nests during 2004–2007, two during egg laying, 28 during incubation, and 12 with nestlings. Mean clutch size was 3.17 eggs (n = 29), the incubation period was 13 days, and nestlings remained in the nest for 12 days before fledgling. Bay-capped Wren-Spinetails are socially monogamous; both males and females develop a brood patch and contribute to incubation, brooding, and provisioning of nestlings. Wren-Spinetails are unique among furnariids as they build an open cup nest with a few presenting a loose domed roof. Breeding success of Bay-capped Wren-Spinetails was higher (total probability of nesting success = 0.508) than other species of sympatric passerines because of low nest predation and high nest survival rates during incubation and nestling rearing stages.

Viruses represent one of the major threats for the strawberry industry, causing important economic losses, and their control is based mainly on rigorous certification programs to maintain clean planting stocks; these viruses include SCV (Strawberry Crinkle Virus) the causal agent of a quarantine disease of major economic importance. All strawberry cultivars are susceptible to SCV infection but due to the absence of obvious symptoms, the detection is limited to indexing on strawberry indicator plants, an expensive and time-consuming procedure. The aim of this study is to develop a sensitive, reliable and fast RealTime assay, based on TaqMan chemistry and amplifying a portion of the conserved L gene sequence coding for RNA-dependent RNA polymerase (RdRp). As strawberry is a difficult host for virus detection, an evaluation of the efficiency of 4 different RNA extraction methods and a comparison between conventional nested-PCR and RealTime assay were carried out. RealTime detection of SCV in several symptomatic and asymptomatic strawberry plants, field collected, indexed on strawberry indicator plants and/or maintained under CAV's screen-houses, proved highly efficient and reproducible, and its sensitivity was as high as the nested PCR assay. This new protocol represents an improvement on the existing analytical methods and could be used as a reliable diagnostic procedure for routine tests in certification and control programs.

The increasing popularity of outdoor recreational activities in recent years has resulted in elevated human disturbance of waterbird communities. Anthropogenic disturbance is defined as any human activity that constitutes a stimulus sufficient to disrupt normal activities and/or distribution of animals relative to the situation in the absence of that activity. The goals of this study were (1) to quantify changes in habitat use by waterbirds caused by the proximity of people’s activities to the shoreline, in Los Padres Lagoon Reserve (Argentina), (2) to evaluate differential responses of waterbird groups caused by this human disturbance, and (3) to propose management guidelines to improve waterbird conservation in Pampas lagoon. We performed bird surveys monthly in areas with high levels of disturbance (HD areas) and with no or low levels of disturbance (LD areas) via recreational activities during days with (weekends) and without (weekdays) presence of people close to the lagoon. We recorded 34 bird species using the lagoon. The bird richness and abundance in HD areas was higher in days without recreational activities, conversely, in LD areas we found no differences in these parameters between days. Waders were found to be the group most vulnerable to disturbance, since these birds were only recorded in HD areas in days without human activity. Podicipedidae, as a group were less affected by recreational activities. We detected changes in the waterbird assembles and structure in relation to the presence of people on the shoreline. Our results in this study suggest direct effects of recreational activities on the habitat use of waterbirds. The buffer area defined by the current Reserve management strategy is working properly, and the impact of recreational activities on transitional area has only instantaneous effects on waterbirds because they return to that area in absence of disturbance. However, it should be considered that we only estimated the response to short-term effects of these activities on the waterbird community. Further studies should assess long-term effects.

Objective: This study presents clinical and radiological outcome of a percutaneous technique for the correction of hallux valgus and lesser toe deformities. Methods: We present a 36 months follow-up series of 32 patients who have been treated with the Reverdin- Isham osteotomy for the correction of hallux valgus, and a percutaneous technique for the correction of lesser toes deformities and metatarsalgia. Clinical outcome data were recorded with the AOFAS score. Radiologic evaluation consisted of weight bearing (AP, lateral and Walter-Muller) views pre and postoperatively at 6 weeks, 3, 6, 12 and 36 months after surgery. Results: At three year follow-up, the mean difference of the HVA was 9.2 (p

The burrowing crab,Chasmagnathus granulatus, is the dominant benthic macroinvertebrate of southwest Atlantic salt marshes and tidal flats, having strong ecosystem engineering effects by direct and indirect effects on soil, vegetation, invertebrates, fishes, and birds. Vegetation structure is a main component for bird habitat selection, since greater habitat complexity generates higher niche diversity. This environmental complexity can be modified by species interactions or disturbance events (i.e., by crabs), in turn modifying the associated community. The bird species of salt marshes of the southwest Atlantic are highly dependent on these ecosystems. We assessed the effects of the burrowing crab on the structure of the cordgrass,Spartina densiflora, marshes, and how these changes affected the composition and diversity of birds. This study was conducted at the Mar Chiquita Coastal Lagoon, Argentina (37°40′S, 57°23′W).S. densiflora marshes were classified within three categories: marshes without burrowing crabs, marshes with low burrow densities, and marshes with high burrow densities. We made vegetation and bird surveys during spring beforeS. densiflora produce seeds and in summer when seeds are abundant. We also manipulated inflorescence (i.e., spike) density (a variable affected by crabs) to assess the effect on bird habitat use. The highest inflorescence density ofS. densiflora and highest bird diversity were recorded at low burrow densities. Cover and height ofS. densiflora were negatively correlated with burrow density. The number of bird species and the number of red-capped wren-spinetail,Spartonoica maluroides, were negatively related with cover and height ofS. densiflora. Experiments confirmed that granivorous species used areas with higher spike densities, a variable related to crab burrow density. Burrowing crabs directly and indirectly affect the cover, height, spike density, and morphologic characteristic of seeds ofS. densiflora marshes. These effects indirectly affect the bird community that uses these marshes, being negatively related toS. maluroides and total bird abundance, and positively related to bird diversity.

Trypanosoma cruzi, the causing agent of Chagas disease, leads to an activation of the immune system in congenitally infected infants. In this study, we measured a set of cytokines/chemokines and the levels of parasitemia by quantitative PCR in the circulation of neonates born to T. cruzi-infected mothers to evaluate the predictive value of these mediators as biomarkers of congenital transmission.We conducted a retrospective cohort study of 35 infants with congenital T. cruzi infection, of which 15 and 10 infants had been diagnosed by detection of parasites by microscopy in the first and sixth month after delivery, respectively, and the remaining 10 had been diagnosed by the presence of T. cruzi-specific Abs at 10-12 mo old. Uninfected infants born to either T. cruzi-infected or uninfected mothers were also evaluated as controls. The plasma levels of IL-17A, MCP-1, and monokine induced by IFN-γ were increased in infants congenitally infected with T. cruzi, even before they developed detectable parasitemia or seroconversion. Infants diagnosed between 6 and 12 mo old also showed increased levels of IL-6 and IL-17F at 1 mo of age. Conversely, infants who did not develop congenital T. cruzi infection had higher levels of IFN-γ than infected infants born to uninfected mothers. Monokine induced by IFN-γ, MCP-1, and IFN-γ production induced in T. cruzi-infected infants correlated with parasitemia, whereas the plasma levels of IL-17A, IL-17F, and IL-6 were less parasite load dependent. These findings support the existence of a distinct profile of cytokines and chemokines in the circulation of infants born to T. cruzi-infected mothers, which might predict congenital infection. Fil: Volta, Bibiana Julieta. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”. Instituto Nacional de Parasitología "Dr. Mario Fatala Chaben”; Argentina Fil: Bustos, Patricia Laura. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”. Instituto Nacional de Parasitología "Dr. Mario Fatala Chaben”; Argentina Fil: Cardoni, Rita Liliana. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”. Instituto Nacional de Parasitología "Dr. Mario Fatala Chaben”; Argentina Fil: de Rissio, Ana María. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”. Instituto Nacional de Parasitología "Dr. Mario Fatala Chaben”; Argentina Fil: Laucella, Susana Adriana. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”. Instituto Nacional de Parasitología "Dr. Mario Fatala Chaben”; Argentina Fil: Bua, Jacqueline Elena. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”. Instituto Nacional de Parasitología "Dr. Mario Fatala Chaben”; Argentina

Dominant individuals often grow faster than subordinates because they gain a greater share of important resources. However, dominants should also strategically adjust their growth rates, relative to the size of subordinates, if this improves their reproductive success. Here, we show that individuals in breeding pairs of the coral-dwelling fish Gobiodon histrio regulate their growth to reduce the size difference between partners. In pairs where one individual was larger than the other, the smaller individual increased its growth rate and the larger individual decreased its growth rate, compared to individuals in size-matched pairs. The reproductive success of breeding pairs is limited by the size of the smallest individual in the pair. Therefore, it appears that the larger individual trades-off its own growth against that of the smaller individual, thereby improving the reproductive success of both individuals in the pair. This demonstrates a remarkable ability of individuals to strategically adjust their body size to suit the local social environment, and reveals a novel mechanism for size-assortative mating.

Mother-to-child transmission of intracellular parasites could be related to the production of immunoregulatory cytokines. The levels of gamma interferon (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha) and lnterleukin (IL)-10 were evaluated during pregnancy in sera of women chronically infected with Trypanosoma cruzi that delivered infected or non-infected children. The levels of IL-10 increased in both, women only pregnant and only infected, compared to non-infected non-pregnant women. However, in pregnant women chronically infected with T. cruzi, IL-10 did not increase significantly, neither in the mothers of infected nor in the mothers of non-infected children. The levels of the inflammatory cytokine TNF-alpha were not affected in normal pregnancy but increased in the infected mothers of non-infected children. The levels of IFN-gamma did not increase in the groups studied, indicating that the production of this pro-inflammatory cytokine was controlled, even when the levels of IL-10 did not increase, as in pregnant women chronically infected with T. cruzi.

Chagas congenital infection is an important health problem in endemic and non-endemic areas in which Trypanosoma cruzi-infected women can transmit the parasite to their offspring. In this study, we evaluated the antibody levels against the T. cruzi Shed Acute Phase Antigen (SAPA) in 91 binomial samples of seropositive pregnant women and their infected and non-infected children by ELISA. In 70 children without congenital T. cruzi transmission, the titers of anti-SAPA antibodies were lower than those of their seropositive mothers. In contrast, 90.5% of 21 congenitally infected children, at around 1 month of age, showed higher anti-SAPA antibody levels than their mothers. Subtracting the SAPA-ELISA mother OD value to the SAPA-ELISA child OD allowed efficient detection of most T. cruzi congenitally infected children immediately after birth, when total anti-parasite antibodies transferred during pregnancy are still present in all children born to seropositive women. A positive correlation was observed between parasitemia levels in mothers and infants evaluated by quantitative DNA amplification and anti-SAPA antibody titers by ELISA. As SAPA serology has proved to be very efficient to detect T. cruzi infection in mother-child binomial samples, it could be of extreme help for early diagnosis of newborns, in maternities and hospitals where DNA amplification is not available. This prompt diagnosis may prevent drop out of the long-term follow-up for future diagnosis and may ensure early trypanocidal treatment, which has proved to be efficient to cure infants with congenital Chagas disease. Fil: Volta, Bibiana Julieta. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud. Instituto Nacional de Parasitología; Argentina Fil: Russomando, Graciela Mabel. Universidad Nacional de Asunción; Paraguay Fil: Bustos, Patricia Laura. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud. Instituto Nacional de Parasitología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Scollo, Karenina. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud. Instituto Nacional de Parasitología; Argentina Fil: de Rissio, Ana María. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud. Instituto Nacional de Parasitología; Argentina Fil: Sánchez, Zunilda. Universidad Nacional de Asunción; Paraguay Fil: Cardoni, Rita Liliana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud. Instituto Nacional de Parasitología; Argentina Fil: Bua, Jacqueline Elena. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud. Instituto Nacional de Parasitología; Argentina. Universidad Abierta Interamericana; Argentina

Gamma-interferon (IFN-gamma) production, the hallmark of the Th1 immune response, has been shown to play a central role in the resistance to Trypanosoma cruzi infections, in particular when produced in the very early acute infection. BALB/c mice infected with T. cruzi, Tulahuén strain, reach high parasitemias during the acute phase, and their spleen cells release IFN-gamma in the second week of the infection, while those of the resistant C3H strain produce the cytokine earlier, at 2 days post-infection (pi). We studied in the spleen cells supernatants of infected BALB/c and C3H mice, the spontaneous production of cytokines involved in the induction, interleukin (IL)-18 and IL-12 p70, as well as in the downregulation, IL-13 and IL-10, of the Th1 immune response. We found that, at 2 days pi, only C3H mice produced IL-18, while IL-12 p70 was detected in both mouse strains. Moreover, at this time pi splenocytes from BALB/c mice spontaneously produced high amounts of IL-13. At 14 days pi, despite the increased levels of IL-13 and IL-10 detected in C3H mice, they still showed high concentrations of IL-18 and IL-12 p70. In contrast, spleen cells from BALB/c mice did not secrete IL-18, IL-12 p70 and IL-13 at this time pi, but produced higher amounts of IL-10 than C3H mice. Non of these cytokines was found increased in the cell supernatants of chronically infected mice. The addition of lipopolysaccharide (LPS) or Concanavalin A (Con A) to the cell cultures did not enhance the production of IL-18 and IL-12 at the time points tested. On the other hand, at 21 days pi, when parasitemia peaked, an inhibition of both the LPS induced IL-10 release and the IL-13 production upon Con A stimulation was observed in C3H, but not in BALB/c mice. We did not find an increase of IL-18, IL-10, or IL-12 p70 in the serum of the infected mice, despite the high seric IL-12 p40 concentrations reached during the infection. The data show that the different kinetics of the production of these cytokines in the spleen of both mouse strains could have a key role in the in vivo regulation of IFN-gamma production. In these experimental models, early IFN-gamma release and thus resistance to T. cruzi infection, could be related to the combined effect of both IL-18 and IL-12p70 in the absence of IL-13.

Latent mosaic is a widely distributed disease of peach caused by peach latent mosaic viroid (PLMVd). Some strains induce mosaic symptoms on leaves, delay in leaf emergence, flowering and maturity, calico and malformation of fruits with cracked sutures and enlarged pits, bud necrosis and, in more severe cases, early decline of the trees. To prevent the use of infected propagating material, international certification schemes provide for the absence of PLMVd from peach mother plants. Until a few years ago, detection of PLMVd was mainly based on cross-protection tests on GF305 peach seedlings. Recently, improved knowledge of the PLMVd genome and optimization of molecular techniques applied to the diagnosis of plant pathogens have stimulated studies aimed at setting up diagnostic methods that are sensitive, reliable and less time-consuming. A comparative trial among different Italian institutions has been set up to compare the sensitivity, specificity and reproducibility of the currently available diagnostic techniques that are routinely applied for the detection of PLMVd in peach germplasm. In this paper, we compare the results obtained in the diagnosis of PLMVd performed with biological indicators and molecular techniques (‘spot-blot’ and ‘tissue-blot’ hybridization using a non-radioactive probe and reverse transcriptase-polymerase chain reaction) starting from different organs (leaves, buds and bark) collected from infected and healthy peach plants and using two total nucleic acid extraction methods. On the basis of the results obtained, we suggest that a tissue-blot hybridization assay using a PLMVd cRNA digoxigenin-labelled probe and starting from bud tissue is a reliable diagnostic method for inclusion in quarantine or certification protocols that require the absence of PLMVd from propagating material of peach.

One of the most extensive grassland ecosystems in the Neotropics is located in the southeastern South America region. Here grasslands once dominated the Pampas but these are now mostly reduced to a mosaic of patches with different land-uses, largely croplands and pasturelands. Native Cortaderia selloana grasslands are widely distributed in the eastern Pampas region but relatively little is known of the bird assemblage inhabiting this habitat. We studied this bird assemblage addressing the overall richness and presence of species of conservation concern, seasonal variation in species composition, breeding phenology and the importance of C. selloana habitats for birds. Species richness was high: 54 species belonging to 22 families, including six species of conservation concern: three classified as vulnerable to extinction (dot-winged crake Porzana spiloptera, black-and-white monjita Xolmis dominicanus and pampas meadowlark Sturnella defilippii), and three near threatened (greater rhea Rhea americana, bay-capped wren-spinetail Spartonoica maluroides and Hudson’s canastero Asthenes hudsoni). The highest species richness was observed in spring and summer, with peaks in spring and marked drops in autumn. The insectivore guild was the most numerous throughout the year. We identified 21 nesting species, four of which were of conservation concern, with a peak of reproductive activity during November. Owing to the high species richness and of the large number of threatened species, C. selloana grasslands should be considered of outstanding conservation priority in the Pampas region Los pastizales del sureste de Sudamérica conforman uno de los mayores ecosistemas de pastizales en el Neotrópico. La vastedad de pastizales que una vez dominó la región actualmente es un mosaico de parcelas sometidas a diferentes usos de la tierra, especialmente destinadas a cultivos pastos. Los pastizales nativos de Cortaderia selloana se encuentran ampliamente distribuidos al este de la región pampeana; sin embargo, a pesar de su dominancia, es relativamente poco lo que se conoce sobre el ensamble de aves que utilizan este hábitat. Hemos estudiado el ensamble de aves que utilizan los pastizales de C. selloana, describiendo la riqueza total, la presencia de especies de interés de conservación, la variación estacional de las especies, la fenología de la reproducción y la importancia de este hábitat para las aves pampeanas. Encontramos una alta riqueza específica conformada por 54 especies pertenecientes a 22 familias, entre las cuales hay seis especies con interés de conservación: la polluela overa Porzana spiloptera, la loica pampeana Sturnella defilippii y la monjita dominicana Xolmis dominicanus, clasificadas como vulnerables a la extinción, y el ñandú común Rhea americana, el canastero enano Spartonoica maluroides y el canastero pampeano Asthenes hudsoni como cercanos a la amenaza. Los mayores valores de riqueza fueron observados durante la primavera y el verano, con picos de abundancia en primavera, y un marcado descenso en otoño. El gremio insectívoro fue dominante en abundancia durante todo el año. Identificamos 21 especies anidando, cuatro de ellas con interés de conservación. La mayor actividad de reproducción se concentró durante noviembre. Debido a su elevada riqueza específica, y particularmente por el número de especies de interés de conservación, el pastizal de C. selloana debería ser destacado por su prioridad de conservación para la región pampeana. Fil: Pretelli, Matías Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Isacch, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Cardoni, Daniel Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina

Background According to the Chagas congenital transmission guides, the diagnosis of infants, born to Trypanosoma cruzi infected mothers, relies on the detection of parasites by INP micromethod, and/or the persistence of T. cruzi specific antibody titers at 10–12 months of age. Methodology and Principal Findings Parasitemia levels were quantified by PCR in T. cruzi-infected children, grouped according to the results of one-year follow-up diagnosis: A) Neonates that were diagnosed in the first month after delivery by microscopic blood examination (INP micromethod) (n = 19) had a median parasitemia of 1,700 Pe/mL (equivalent amounts of parasite DNA per mL); B) Infants that required a second parasitological diagnosis at six months of age (n = 10) showed a median parasitemia of around 20 Pe/mL and 500 Pe/mL at 1 and 6 months old, respectively, and C) babies with undetectable parasitemia by three blood microscopic observations but diagnosed by specific anti - T. cruzi serology at around 1 year old, (n = 22), exhibited a parasitemia of around 5 Pe/mL, 800 Pe/mL and 20 Pe/mL 1, 6 and 12 month after delivery, respectively. T. cruzi parasites were isolated by hemoculture from 19 congenitally infected children, 18 of which were genotypified as DTU TcV, (former lineage TcIId) and only one as TcI. Significance This report is the first to quantify parasitemia levels in more than 50 children congenitally infected with T. cruzi, at three different diagnostic controls during one-year follow-up after delivery. Our results show that the parasite burden in some children (22 out of 51) is below the detection limit of the INP micromethod. As the current trypanocidal treatment proved to be very effective to cure T. cruzi - infected children, more sensitive parasitological methods should be developed to assure an early T. cruzi congenital diagnosis., Author Summary Chagas or American Trypanosomiasis is a disease that affects around 8–10 million people in Latin America, and can be transmitted by congenital infection. In areas where the vector insect and blood transfusions are controlled, this mode of transmission is epidemiologically important, as 15,000 Trypanosoma cruzi-infected children are born each year. Parasitological methods are available to detect T. cruzi but our results suggest that more sensitive diagnostic methods should be developed, as not all newborns have high parasite burdens to be detected by microscopy. If congenital infected babies are promptly diagnosed after delivery they can be cured, as the trypanocidal treatment is very effective in children and with no significant side effects.

The relationship between parasite burden and vertical transmission of Trypanosoma cruzi was studied in pairs of chronically infected women and their children in a non-endemic area. Parasitemia was quantified by quantitative polymerase chain reaction (qPCR) in the peripheral blood amplifying a nuclear T. cruzi DNA and expressed as equivalent amounts of CL Brener parasites DNA per ml (eP/ml). Similar levels of parasitemia were found in non-transmitting pregnant women and in non-pregnant women: 1.8 ± 0.5 and 1.5 ± 0.7 eP/ml, respectively. In women pregnant with infected children parasitemia was 11.0 ± 2.7 eP/ml (n=20). In 12 of their neonates the infection was detected by microscopic observation of the parasites in peripheral blood in the 1(st) month of age. These children had variable levels of parasitemia (13,000 ± 7000 eP/ml), that were about 600-fold higher than that found in their mothers. To our knowledge, this is the first quantitative evaluation of parasitemia in these three groups of women and in their congenitally infected children. These parasite quantifications could be a basis to plan the control of mother-to-child transmission of T. cruzi.

Eutrophication increases the biomass of opportunistic green macroalgae that covers intertidal zones, and macroalgal blooms may affect the intertidal invertebrate community and predation of invertebrates by shorebirds. In San Antonio Bay, Argentina, eutrophication from the discharge of wastewater from a coastal town produces periodic macroalgal blooms. Our aim was to assess if macroalgal blooms affect the foraging behaviour and diet of the American Oystercatcher (Haematopus palliatus). A macroalgal transplant experiment was performed in order to evaluate how epifaunal species respond to a macroalgal canopy. The availability of prey for Oystercatchers, and their foraging behaviour and diet, were analysed in two paired channels with different nutrient loadings. Oystercatchers generally ate the most profitable prey and avoided prey with a profitability value lower than the mean rate of energy intake. During the macroalgal blooms, Oystercatchers avoided two prey species with high profitability values, shifting their foraging strategy and feeding onto a suboptimal prey but with a high encounter rate. Our results suggest that nutrient loadings and the macroalgal blooms that they generate have effects on the diet and foraging behaviour of Oystercatchers, which results in an increase of the average rate of energy intake of Oystercatchers foraging along the channel subject to a macroalgal bloom. Fil: García, Germán Oscar. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Biología; Argentina Fil: Isacch, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Biología; Argentina Fil: Gómez Laich, Agustina Marta. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico; Argentina Fil: Albano, Mariano Javier. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Biología; Argentina Fil: Favero, Marco. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Biología; Argentina Fil: Cardoni, Daniel Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Biología; Argentina Fil: Luppi, Tomas Atilio. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Biología; Argentina Fil: Iribarne, Oscar Osvaldo. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Biología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Most congenital transmissions of Trypanosoma cruzi are not detected. As the levels of mediators regulating the immune response might be different in the absence or in the presence of transmission, we explored the levels of tumor necrosis factor (TNF) and soluble TNF receptors TNF-R1 and -R2 in T. cruzi-infected pregnant women and the neonates. We previously found that the circulating levels of TNF were higher in non-transmitting than in transmitting pregnant women. This observation has now been extended to the spontaneous release of TNF by peripheral blood leukocytes (PBLs) that was also higher in non-transmitting than in transmitting pregnant women. As their mothers, non-infected neonates had higher circulating levels of TNF than congenitally infected children. The circulating levels of sTNF-R1 increased in non-transmitting and transmitting mothers and in infected and non-infected neonates. The circulating levels of sTNF-R2 were approximately 60% higher in infected than in non-infected neonates (1,635 +/- 101 and 1,027 +/- 100 pg/mL, respectively) and remained higher at 1 year of age. This important increase, only observed in infected neonates, could be useful to orientate to the presence of vertical transmission of T. cruzi infection.

Cellular populations involved in resistance against T. cruzi infection were characterized from mice chronically infected with the parasite. Mice transfused with spleen cells (SC), nylon-wool-non-adherent spleen cells (NWNA) or sera from mice chronically infected with T. cruzi , showed an enhanced resistance against challenge with the parasite. The protective activity of NWNA but not of SC was completely abrogated by treatment with anti-Thyl.2 monoclonal antibodies (mAb) and complement (C). Pretreatment of NWNA cells from chronically infected mice with either anti-L3T4 or anti-Lyt 2.2 mAb partially reduced the transfer of resistance. When both L3T4 + and Lyt2.2 + cells were depleted from NWNA populations, transfer of resistance was abolished. These results appear to indicate that L3T4 + , Lyt2.2 + T cell subsets and non-T cells are involved in the immunity to T. cruzi .

TRYPANOSOMA cruziinduces inflammatory reactions in several tissues. The production of prostaglandin F2α, 6-keto-prostaglandin F1αand thromboxane B2, known to regulate the immune response and to participate in inflammatory reactions, was studied in mice experimentally infected withT. cruzi.The generation of nitric oxide (NO), which could be regulated by cyclooxygenase metabolites, was also evaluated. In the acute infection the extension of inflammatory infiltrates in skeletal muscle as well as the circulating levels of cyclooxygenase metabolites and NO were higher in resistant C3H mice than in susceptible BALB/c mice. In addition, the spontaneous release of NO by spleen cells increased earlier in the C3H mouse strain. In the chronic infections, the tissue inflammatory reaction was still prominent in both groups of mice, but a moderate increase of thromboxane B2concentration and in NO released by spleen cells was observed only in C3H mice. This comparative study shows that these mediators could be mainly related to protective mechanisms in the acute phase, but seem not to be involved in its maintenance in the chronicT. cruziinfections.

T and NK cells play a key role in resistance to Trypanosoma cruzi infections, mainly through IFN-gamma production. The expression of T and NK cells surface markers was studied in NWNA spleen cells of resistant C3H and susceptible BALB/c mice that release IFN-gamma in the early and late acute infection, respectively. In the progressively enlarged spleens, we found: (a) an increased percentage and number of NK blast cells as early as at 2 days post-infection (pi), (b) an enrichment of T and NK cells, in both the total and blast populations, during the late acute phase. At 17 days pi, there was also an accumulation of TCR- alphabeta+DX5+, NKT cells, mainly in resistant mice. At 21 days pi, the enrichment of NK cells ceased, while spleen cells and the T cell compartment continued their expansion. In the chronic stage, TCR-alphabeta+ blasts were expanded in both mouse strains, but NK blasts increased only in BALB/c that, unlike C3H mice, release IFN-gamma. As T and NK cell proliferation is not always associated to IFN-gamma release the experimental downregulation of their expansion to avoid tissue damage could be explored.

The changes in the T cell subsets of the Peyer's patches and the thymus were analyzed in BALB/c mice infected with Trypanosoma cruzi, Tulahuen strain. During the acute stage of the infection both lymphoid organs drastically reduced their cellularity. This was mainly due to the decrease in the immature CD4 + CD8 + T cell population in the thymus and in both T and B cells in the Peyer's patches. In the acute infection, few Peyer's patches were found and the histological studies revealed a depletion of the thymic-dependent areas, paralleling the decreased number of cells expressing CD4 and αβ T cell receptor. After 14 weeks, in the late stage of the infection, the cellularity and the levels of the T cell subsets studied returned to values similar to those of noninfected mice.

The release of reactive oxygen intermediates (ROI), mediators of inflammatory reactions, was evaluated in murine Trypanosoma cruzi infection. In acutely infected BALB/c mice, spleen cells were stimulated, either with epimastigote or trypomastigote forms of the parasite, and the effect was enhanced by serum from infected mice. Only opsonized parasites triggered the release of ROI by normal mouse cells and this response was several times lower than in infected mice. This seems to indicate that cells from acutely infected mice reacted to T. cruzi and that neither parasites nor serum factors blocked the release of ROI. During the acute stage of the infection, both the parasitemia and the release of ROI by spleen cells were higher in BALB/c than in C3H mice (ROI generated in response to a phagocytic stimulation was 12 and 3 times the normal levels, respectively). In addition, in BALB/c mice infected with different numbers of parasites, the production of ROI was related to parasitemia. On the other hand, during the chronic stage of the infection, the inflammatory reaction in myocardium was greater in C3H than in BALB/c mice, and the increase in ROI production was 30% and 100% above the normal levels in BALB/c and C3H mice, respectively. This suggests that the increased ROI production paralleled the parasite burden in the acute phase, and could be related to inflammatory processes after the control of the parasitemia.

OBJECTIVE: The presence of autoantibodies with adrenergic and cholinergic activity, capable of triggering neurotransmitter receptor-mediated effects, has been associated with pathogenesis in T. cruzi-infected hosts. The goal of this study was to investigate the production of anti-M2 muscarinic receptor autoantibodies (Anti-M2R AAbs) as well as the IFN-γ profile in children at the early stage of Chagas disease, and to examine whether trypanocidal chemotherapy with benzonidazole (BZ) could modify both response patterns. METHODS: This study comprised 30 T. cruzi-infected children (mean age: 13.8 years) and 19 uninfected controls (mean age: 12.7 years). Infected patients were treated with BZ and followed-up. Blood samples collected at diagnosis-T0, end of treatment-T1, and six months later-T2 were analysed by ELISA for detection of Anti-M2R AAbs and circulating levels of IFN-γ. RESULTS: At T0, anti-M2R AAbs were demonstrated in 56.7% of T. cruzi-infected patients, whereas uninfected controls were 100% negative. The average age of Anti-M2R AAbs(+) patients was higher than that from negative population. Infected children also displayed significantly stronger serum IFN-γ responses than controls. Upon BZ treatment, a significant linear decreasing trend in Anti-M2R AAb reactivity was recorded throughout the follow-up, with 29.7-88.1% decrease at T2. IFN-γ circulating levels also declined by T2. CONCLUSION: Anti-M2R AAbs and IFN-γ raise early during chagasic infection in children and are downmodulated by BZ therapy. These findings reinforce the usefulness of early BZ treatment not only to eliminate the parasite but also to reduce potentially pathogenic immune responses.

Effector mechanisms of resistance exerted by T cells from BALB/c mice chronically infected with Trypanosoma cruzi, Tulahuen strain, were studied. Spleen cells from chronically infected mice (Chro-SC) prestimulated with heat-killed trypomastigotes (HKT) and/or IL-2 destroyed PHA-labeled p-815 mastocytoma cells, HKT-pulsed macrophages, and normal peritoneal macrophages. However, HKT-stimulated Chro-SC did not affect the infectivity of free bloodstream forms of the parasite. Upon HKT stimulation, Chro-SC or their culture supernatant activated peritoneal macrophages for the destruction of intracellular amastigotes. The effect was abolished after Thy 1.2+ cell depletion. The addition of Cyclosporin A (CyA), which blocks T-cell activation, during HKT-stimulation of Chro-SC, diminished their ability to activate the trypanocidal activity of macrophages. CyA also inhibited the production of both macrophage-activating factors and interferon-gamma by HKT-stimulated Chro-SC. CyA administration to recipients of nylon-wool nonadherent spleen cells from chronically infected mice inhibited their adoptively acquired resistance against T. cruzi, suggesting that the conferred resistance depended on the effect of specifically activated cells. When administered during the chronic stage of the infection, CyA abrogated the antigen-specific delayed type hypersensitivity response but increased the levels of anti-T. cruzi IgG antibodies. Neither parasitemia, tissular parasitism in myocardium or skeletal muscle, nor mortality were detected after CyA treatment, suggesting the presence of a CyA nonsensitive mechanism(s) in the control of T. cruzi during the chronic phase of the infection.