Haemonchus contortus con resistencia múltiple a los antihelmínticos de corta y larga acción y consideraciones sobre el impacto sanitario-productivo de este fenómeno en una majada de ovinos lecheros de la provincia de Santa Fe

Entre Octubre y Noviembre del 2012 se realizaron evaluaciones sobre la actividad de cuatro antihelmínticos con diferente mecanismo de acción (levamisol, ivermectina, closantel y febendazol) en ovinos lecheros naturalmente parasitados por nematodes gastrointestinales en un establecimiento del área central de Santa Fe. Las determinaciones de susceptibilidad o resistencia se realizaron a través del test de reducción en el conteo de huevos (TRCH) y como animales experimentales se utilizaron 40 corderos destetados de la raza Pampinta de aproximadamente dos meses y medio de edad. Estos animales fueron asignados a cuatro grupos de tratamientos de diez animales cada uno en función del número de huevos de nematodes por gramo de heces o hpg (p>0,005). El TRCH se determinó estableciendo el porcentaje de reducción observado en el promedio del hpg en las muestras de materia fecal pre y post tratamiento de los mismos animales. Los resultados indicaron la presencia predominante de Haemonchus contortus con resistencia múltiple a ivermectina, febendazol y clo-santel así como de Nematodirus spp con resistencia a las dos primeras de estas drogas. El levamisol presentó una eficacia muy alta (> al 99 %) para controlar cualquiera de estos aislamientos. Ante la presencia de resistencia con los otros antihelmínticos, todos los corderos experimentales fueron re-tratados con levamisol observándose en este segundo TRCH una eficacia similar al primero. No obstante, debido a su baja actividad larvicida y su corta acción, aproximadamente 30 días posteriores al tratamiento de todos los corderos con levamisol se produjo un severo caso de haemonchosis con elevados índices de mortalidadBetween October and November 2012 a field trial was carried out in dairy sheep naturally parasi-tized by gastrointestinal nematodes in central Santa Fe (Argentina) to investigate the activity of four anthelmintic with different mechanism of action (levamisole, ivermectin, fenbendazole and closantel). The status of susceptibility or resistance was performed through the test of reduction in egg count (TRCH) on 40 Pampinta lambs about two and half months old. These lambs were assigned to four treatment groups of ten animals each according to the number of nematode eggs per gram of feces or hpg (p> 0.005). TRCH was determined by establishing the percentage reduction in the average observed epg in fecal samples before and after treatment of the same animals. The results indicated the predominance of Haemonchus contortus with multiple resistance to ivermectin, febendazole and closantel as well as Nematodirus spp with resistance to the first two of these drugs. Levamisole showed a very high efficacy (> 99%) to control any of these isolates. Due to the presence of Haemonchus spp with resistance to ivermectin, febendazole and closantel, all experimental lambs were re-treated with levamisole and similar efficacy in the TRCH was observed in this second trial. However, due to its low larvicidal activity and short action, approximately 30 days after levamisol treatment, a severe outbreak of acute haemonchosis with high mortality rates was observedEEA RafaelaFil: Muchiut, Sebastian. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias; ArgentinaFil: Mildenberger, M. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias; ArgentinaFil: Pujato, Andrés. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias; ArgentinaFil: Anziani, Oscar Sergio. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentin

Leuconostoc mesenteroides and Leuconostoc pseudomesenteroides bacteriophages: Genomics and cross-species host ranges

Unveiling virus-host interactions are relevant for understanding the biology and evolution of microbes globally, but in particular, it has also a paramount impact on the manufacture of fermented dairy products. In this study, we aim at characterizing phages infecting the commonly used heterofermentative Leuconostoc spp. on the basis of host range patterns and genome analysis. Host range of six Leuconostoc phages was investigated using three methods (efficiency of plaquing, spot and turbidity tests) against Ln. mesenteroides and Ln. pseudomesenteroides strains. Complete genome sequencing from four out of the six studied Leuconostoc phages were obtained in this work, while the remaining two have been sequenced previously. According to our results, cross-species host specificity was demonstrated, as all phages tested were capable of infecting both Ln. pseudomesenteroides and Ln. mesenteroides strains, although with different efficiency of plaquing (EOP). Phage adsorption rates and ability of low-EOP host strains to propagate phages by crossing the Leuconostoc species' barrier confirm results. At the genome level, phages CHA, CHB, Ln-7, Ln-8 and Ln-9 revealed high similarity with previously characterized phages infecting mostly Ln. mesenteroides strains, while phage LDG was highly similar to phages infecting Ln. pseudomesenteroides. Additionally, correlation between receptor binding protein (RBP) and host range patterns allowed us to unveil a finer clustering of Leuconostoc phages studied into four groups. This is the first report of overlapped phage host ranges between Leuconostoc species.This work was supported by the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET; Project PIP 112-201201-00046; Argentina), the Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT; Project PICT 2010-0138; Argentina) and the Universidad Nacional del Litoral (UNL, Project CAI+D PI 501 201101 00039 LI; Argentina). S.A.P. was the recipient of an international scholarship awarded by BEC.AR (Becas de formación en el exterior en Ciencia y Tecnología, Presidencia de la Nación, Argentina). M.M.G. thanks Ministerio de Economía y Competitividad (refs. CGL2013-40564-R and SAF2013-49267-EXP), Generalitat Valenciana (ACOMP/2015/133) and Gordon and Betty Moore Foundation (Grant award ref. 5334). F.J.M.M. is funded by the Spanish Ministerio de Economía y Competitividad (BIO2014-53029P) and the European Commission/Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (291815 Era-Net ANIHWA)

Transcription factor motif quality assessment requires systematic comparative analysis [version 2; referees: 2 approved]

Transcription factor (TF) binding site prediction remains a challenge in gene regulatory research due to degeneracy and potential variability in binding sites in the genome. Dozens of algorithms designed to learn binding models (motifs) have generated many motifs available in research papers with a subset making it to databases like JASPAR, UniPROBE and Transfac. The presence of many versions of motifs from the various databases for a single TF and the lack of a standardized assessment technique makes it difficult for biologists to make an appropriate choice of binding model and for algorithm developers to benchmark, test and improve on their models. In this study, we review and evaluate the approaches in use, highlight differences and demonstrate the difficulty of defining a standardized motif assessment approach. We review scoring functions, motif length, test data and the type of performance metrics used in prior studies as some of the factors that influence the outcome of a motif assessment. We show that the scoring functions and statistics used in motif assessment influence ranking of motifs in a TF-specific manner. We also show that TF binding specificity can vary by source of genomic binding data. We also demonstrate that information content of a motif is not in isolation a measure of motif quality but is influenced by TF binding behaviour. We conclude that there is a need for an easy-to-use tool that presents all available evidence for a comparative analysis

Phages of dairy Leuconostoc mesenteroides: Genomics and factors influencing their adsorption

Phages infecting Leuconostoc mesenteroides strains can be overlooked during milk fermentation because they do not slowdown the acidification process. However, they can negatively impact the flavor profile of the final product. Yet, the information about these phages is still scarce. In this work, we investigated diverse factors influencing the adsorption of seven virulent Ln. mesenteroides phages, isolated from blue cheese manufacture in Argentina, to their host cells. The addition of calcium ions was generally necessary to observe complete cell lysis and plaque formation for four of the seven phages, but adsorption was very high even in the absence of this cation for all phages. The temperature barely influenced the adsorption process as it was high within the temperature range tested (0 to 50. °C). Moreover, the kinetics of adsorption were similar on viable and non-viable cells, revealing that phage adsorption does not depend on physiological state of the bacterial cells. The adsorption rates were also high at pH values from 4 to 9 for all Ln. mesenteroides phages. We also analyzed the complete genome sequences of two of these phages. Complete nucleotide analysis of phages Ln-8 and Ln-9 showed dsDNA genomes with sizes of 28.5 and 28.9. kb, and the presence of 45 and 48 open reading frames (ORFs), respectively. These genomes were highly similar to those of previously characterized Φ1-A4 (USA, sauerkraut, fermentation) and ΦLN25 (England, whey), both virulent Ln. mesenteroides phages. A detailed understanding of these phages will lead to better control strategies.Fil: Pujato, Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; ArgentinaFil: Mercanti, Diego Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; ArgentinaFil: Guglielmotti, Daniela Marta. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; ArgentinaFil: Rousseau, Geneviève M.. Laval University; CanadáFil: Moineau, Sylvain. Laval University; CanadáFil: Reinheimer, Jorge Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; ArgentinaFil: Quiberoni, Andrea del Lujan. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; Argentin

Blocking UBE2N abrogates oncogenic immune signaling in acute myeloid leukemia.

Dysregulation of innate immune signaling pathways is implicated in various hematologic malignancies. However, these pathways have not been systematically examined in acute myeloid leukemia (AML). We report that AML hematopoietic stem and progenitor cells (HSPCs) exhibit a high frequency of dysregulated innate immune-related and inflammatory pathways, referred to as oncogenic immune signaling states. Through gene expression analyses and functional studies in human AML cell lines and patient-derived samples, we found that the ubiquitin-conjugating enzyme UBE2N is required for leukemic cell function in vitro and in vivo by maintaining oncogenic immune signaling states. It is known that the enzyme function of UBE2N can be inhibited by interfering with thioester formation between ubiquitin and the active site. We performed in silico structure-based and cellular-based screens and identified two related small-molecule inhibitors UC-764864/65 that targeted UBE2N at its active site. Using these small-molecule inhibitors as chemical probes, we further revealed the therapeutic efficacy of interfering with UBE2N function. This resulted in the blocking of ubiquitination of innate immune- and inflammatory-related substrates in human AML cell lines. Inhibition of UBE2N function disrupted oncogenic immune signaling by promoting cell death of leukemic HSPCs while sparing normal HSPCs in vitro. Moreover, baseline oncogenic immune signaling states in leukemic cells derived from discrete subsets of patients with AML exhibited a selective dependency on UBE2N function in vitro and in vivo. Our study reveals that interfering with UBE2N abrogates leukemic HSPC function and underscores the dependency of AML cells on UBE2N-dependent oncogenic immune signaling states

MOESM3 of The antibody response in the bovine mammary gland is influenced by the adjuvant and the site of subcutaneous vaccination

Additional file 3. Differences in least square means (Log 2 ) of Îą-toxin specific antibody isotype titers and neutralization titers