BUSCADOR RECOLECTA

[EN]Gastrointestinal nematodes (GIN) are a major threat to health and welfare in small ruminants worldwide. Teladorsagia circumcincta is a nematode that inhabits the abomasum of sheep, especially in temperate regions, causing important economic losses. Given that T. circumcincta and microbiome share the same niche, interactions between them and the host are expected. Although it is known that within a sheep breed there are animals that are more resistant than others to infection by GIN, it is not known if the microbiome influences the phenotype of these animals. Under this condition, 12 sheep were classified according to their cumulative faecal egg count (cFEC) at the end of a first experimental infection, 6 as resistant group (RG) and 6 as susceptible group (SG) to T. circumcincta infection. Then, all sheep were experimentally infected with 70,000 L3 of T. circumcincta and at day 7 days post-infection were euthanized. At necropsy, gastric mucosa and gastric content from abomasum were collected to extract bacterial DNA and sequence V3-V4 region from 16S rRNA gene using Ilumina technology. After bioanalysis performed, results showed that α-diversity and β-diversity remained similar in both groups. However, resistant phenotype sheep showed a higher number of bacteria butyrate-fermenting species as Clostridium sensu stricto 1 (abundance in RG: 1.29% and in SG: 0.069%; p = 0.05), and Turicibacter (abundance in RG: 0.31% and in SG: 0.027%; p = 0.07) in gastric content but also Serratia spp in gastric mucosa (abundance in RG: 0.12% and in SG: 0.041%; p = 0.07). A trend towards a significant negative correlation between cFEC and Clostridium sensu stricto 1 abundance in gastric content was detected (r = − 0.537; p = 0.08). These data suggest that microbiome composition could be another factor associated with the development of the resistant phenotype modifying the interaction with the host and the in last instance affecting the individual risk of infection., SI

[EN]Inflammatory bowel disease (IBD) is a complex, chronic, relapsing and heterogeneous disease induced by environmental, genomic, microbial and immunological factors. MCJ is a mitochondrial protein that regulates the metabolic status of macrophages and their response to translocated bacteria. Previously, an acute murine model of DSS-induced colitis showed increased disease severity due to MCJ deficiency. Unexpectedly, we now show that MCJ-deficient mice have augmented tumor necrosis factor α converting enzyme (TACE) activity in the context of chronic inflammation. This adaptative change likely affects the balance between soluble and transmembrane TNF and supports the association of the soluble form and a milder phenotype. Interestingly, the general shifts in microbial composition previously observed during acute inflammation were absent in the chronic model of inflammation in MCJ-deficient mice. However, the lack of the mitochondrial protein resulted in increased alpha diversity and the reduction in critical microbial members associated with inflammation, such as Ruminococcus gnavus, which could be associated with TACE activity. These results provide evidence of the dynamic metabolic adaptation of the colon tissue to chronic inflammatory changes mediated by the control of mitochondrial function., SI

[EN] Constructed wetlands represent a low-cost and highly efficient municipal wastewater treatment alternative, due to their low technological and energy demands. Wetland vegetation releases an amount of carbon to the system, when it is decomposed (winter period). Part of this organic matter could remain in the system, and will be decomposed at very low rates during winter and spring. In this research, a constructed superficial-flow wetland was divided into two equal parts and vegetation (Typha latifolia) was harvested in one of them. The organic load applied to the system was 11.2 gBOD/m2·d. The control of the organic matter was carried out during 141 d (111 d in winter and 30 d in spring). Differences in efficiencies (TSS, BOD and COD), were observed between both types of the wetland, with an important increase in these differences during spring. Vegetation released organic matter to the system, specially suspended and biodegradable matter. BOD and TSS released per dry gram of Typha were 4.24 mgBOD/gTypha and 4.36 mgTSS/gTypha, respectively. Harvest is a recommended practice in systems treating diluted wastewater, especially in productive areas like the Mediterranean. The recommendations can also be applied to a broader geographic area, SI

[EN] The control of gastrointestinal nematodes in livestock is becoming increasingly difficult due to the limited number of available drugs and the rapid development of anthelmintic resistance. Therefore, it is imperative to develop new anthelmintics that are effective against nematodes. Under this context, we tested the potential toxicity of three compounds in mice and their potential anthelmintic efficacy in Mongolian gerbils infected with Haemonchus contortus. The compounds were selected from previous in vitro experiments: two diamine (AAD-1 and AAD-2) and one benzimidazole (2aBZ) derivatives. 2aBZ was also selected to test its efficacy in sheep. In Mongolian gerbils, the benzimidazole reduced the percentage of pre-adults present in the stomach of gerbils by 95% at a dose of 200 mg/kg. In sheep, there was a 99% reduction in the number of eggs shed in faeces after 7 days at a dose of 120 mg/kg and a 95% reduction in the number of worm adults present in the abomasum. In conclusion, 2aBZ could be considered a promising candidate for the treatment of helminth infections in small ruminants, SI, Financial support came from MINECO: RETOS (AGL2016-79813-C2-1R/2R) and MICINN/AEI (PID2020- 119035RB-100). EVG was funded by FPU17/00627, FPU17/05346; VCGA, MAB, MCP and LGP are recipients of Junta de Castilla y León (JCyL) (LE082-18, LE051-18, LE135-19, LE096-20, respectively) and MMV by the Spanish “Ramon y Cajal” Programme (Ministerio de Economía y competitividad; MMV, RYC-2015-18368)

[EN] Vaccination can be an efficient method for the control of paratuberculosis in ruminants. However, the official tuberculosis control tests cross-interfere with the animals vaccinated against paratuberculosis. In order to test and compare new antigens that could solve this problem, the production of interferon-gamma (IFN-γ) in peripheral blood at different post-vaccination days in experimental kids and adult goats, in field conditions, using the avian and bovine purified protein derivative (PPD), the johnin, two peptide cocktails of Mycobacterium bovis (PC-EC and PC-HP) and the antigens VK 055 and VK 067 of Mycobacterium avium subspecies paratuberculosis (Map) has been analyzed in vitro. The non-specific production of IFN-γ was observed after blood stimulation with the PC-EC and PC-HP cocktail in any sample from vaccinated animals, whereas it was detected when bovine PPD was used. These results support the possible use of these new Mycobacterium bovis antigens in the in the differentiation of animals vaccinated against paratuberculosis or infected with tuberculosis by improving the specificity of bovine PPD. In contrast, the two Map antigens tested in this study did not improve the sensitivity of johnin or avian PPD in the detection of vaccinated or Map-infected goats, SI, This research was funded by Ministry of Economy, Industry and Competitiveness through the research project with reference: AGL2012-39818-C02-01 titled “Development of pathogenesis and immunization models in paratuberculosis”. Marcos Royo and Noive Arteche-Villasol were recipients of a predoctoral contract for training of doctors by the Ministry of Economy, Industry and Competitiveness

[ES] El cosechado de vegetación es uno de los puntos controvertidos en relación con el funcionamiento y la gestión de humedales construidos. La cosecha generalmente se evita, en la mayor parte de los lugares, debido a la baja cantidad de nutrientes eliminados en relación con el gasto generado. En determinadas condiciones la cosecha podría tener importancia para reducir la producción de materia orgánica y aumentar la inactivación bacteriana. Se cosechó la mitad de la superficie de un humedal artificial de flujo superficial plantado con Typha y esta mitad fue aislada de la parte no cosechada utilizando una membrana impermeable. Los efluentes de ambas partes (cosechada y no cosechada) fueron comparados en relación con la eliminación de indicadores fecales. Coliformes fecales, E. coli, estreptococos, estafilococos, Clostridium perfringens y bacteriófagos fueron analizados en el influente y en el efluente entre los meses de diciembre de 2004 y abril de 2005. Los resultados mostraron diferencias estadísticamente significativas en varios grupos bacteriales. Desde este artículo proponemos el sistema de cosechado como una técnica de gestión apropiada en condiciones mediterráneas sobre todo para aguas residuales de baja carga contaminante, [EN] Harvest of vegetation is one of the controversial points in relation to operation and management of constructed wetlands. Harvesting is generally avoided in most places due to the low amount of nutrients removed and its associated costs. In some conditions harvesting could be important as it reduces organic matter production and increases inactivation of bacteria. A superficial-flow constructed wetland planted with Typha was half-harvested and isolated from the non harvested part using a impermeable layer. Efluents from harvested and non-harvested parts were compared in relation to faecal indicators removal. Faecal coliforms, E. coli, streptococci, staphylococci, Clostridium perfringens and phages were analyzed in the influent and effluents from december 2004 to april 2005. Results showed statistically significant differences in several bacterial groups. From this paper we propose harvesting as an appropriated management technique in Mediterranean conditions especially for low loaded wastewaters, SI

[ES] Brian Moss es un ecólogo de reconocido prestigio en el campo de la limnología y con un amplio interés en la educación ambiental y en la interacción entre el hombre y su medio. Sus estudios limnológicos han cubierto un amplio rango geográfico, desde los lagos tropicales hasta los del norte de Europa, y a lo largo de su carrera ha realizado numerosos experimentos para estudiar el efecto de factores como la eutrofización, la introducción de peces o el calentamiento, sobre la estructura y el funcionamiento de los lagos. Actualmente centra su interés en la legislación europea relacionada con la calidad del agua, intentando concienciar a la sociedad sobre su correcta aplicación. Su inquietud docente le ha llevado a participar en numerosos cursos con estudiantes internacionales en diversos países del tercer mundo. Se doctoró en 1968 en la Universidad de Bristol y ha sido profesor en las universidades de Michigan (USA), East Anglia y Liverpool (Reino Unido), donde actualmente dirige la Facultad de Ciencias Biológicas. Ha publicado unos 150 artículos y 5 libros, de los cuales el más conocido es Ecology of Freshwaters, Man and Medium, Past to Future. Ha sido el editor del Journal of Ecology durante los años 1980 a 1987. Brian se define a sí mismo como alguien al que no le gustaría que le vieran como un científico del "establishment", sino como alguien que piensa que es necesario llevar a cabo cambios muy importantes en la ciencia, en las instituciones académicas y en la sociedad si queremos que la hidrosfera, y la biosfera que la contiene, retenga los pocos bienes y servicios que aún le quedan. Su reseña no podría terminar sin citar algunas de sus pasiones como tocar el contrabajo y la poesía. Eloy Bécares le hizo esta entrevista, SI

[EN] Porcine reproductive and respiratory syndrome (PRRS) is a viral disease defined by reproductive problems, respiratory distress and a negative impact on growth rate and general condition. Virulent PRRS virus (PRRSV) strains have emerged in the last years with evident knowledge gaps in their impact on the host immune response. Thus, the present study examines the impact of acute PRRS virus (PRRSV) infection, with two strains of different virulence, on selected immune parameters and on the gut microbiota composition of infected pigs using 16S rRNA compositional sequencing. Pigs were infected with a low virulent (PRRS_3249) or a virulent (Lena) PRRSV-1 strain and euthanized at 1, 3, 6, 8 or 13 days post-inoculation (dpi). Faeces were collected from each animal at the necropsy time-point. Alpha and beta diversity analyses demonstrated that infection, particularly with the Lena strain, impacted the microbiome composition from 6 dpi onwards. Taxonomic differences revealed that infected pigs had higher abundance of Treponema and Methanobrevibacter (FDR < 0.05). Differences were more considerable for Lena- than for PRRS_3249-infected pigs, showing the impact of strain virulence in the intestinal changes. Lena-infected pigs had reduced abundancies of anaerobic commensals such as Roseburia, Anaerostipes, Butyricicoccus and Prevotella (P < 0.05). The depletion of these desirable commensals was significantly correlated to infection severity measured by viraemia, clinical signs, lung lesions and immune parameters (IL-6, IFN-γ and Hp serum levels). Altogether, the results from this study demonstrate the indirect impact of PRRSV infection on gut microbiome composition in a strain virulence-dependent fashion and its association with selected immune markers, SI, J. G omez-Laguna is supported by a ‘Ram on y Cajal’ contract of the Spanish Ministry of Economy and Competitiveness (RYC-2014-16735). Hector Arg€uello is supported by the ‘Beatriz Galindo’ Programme from the Spanish Ministry of Education (BEAGAL-18-106). This work was partially supported by the Spanish Ministry of Education and Science (AGL2016-76111-R). Research in the Cotter laboratory is funded by Science Foundation Ireland in the form of a centre grants (APC Microbiome Ireland, Grant Number SFI/12/RC/2273, and Vistamilk, Grant Number SFI/16/RC/3835) and by the European Commission under the Horizon 2020 programme under grant number 818368 (MASTER)

[EN] Bovine paratuberculosis (PTB) is a chronic enteritis caused by Mycobacterium avium subspecies paratuberculosis (Map) that causes a heavy economic impact worldwide. Map infected animals can remain asymptomatic for years while transmitting the mycobacteria to other members of the herd. Therefore, accurate detection of subclinically infected animals is crucial for disease control. In a previous RNA-Seq study, we identified several mRNAs that were overexpressed in whole blood of cows with different PTB-associated histological lesions compared with control animals without detected lesions. The proteins encoded by two of these mRNAs, ATP binding cassette subfamily A member 13 (ABCA13) and Matrix Metallopeptidase 8 (MMP8) were significantly overexpressed in whole blood of animals with focal histological lesions, the most frequent pathological form in the subclinical stages of the disease. In the current study, the potential of sensitive early diagnostic tools of commercial ELISAs, based on the detection of these two biomarkers, was evaluated in serum samples of 704 Holstein Friesian cows (566 infected animals and 138 control animals from PTB-free farms). For this evaluation, infected animals were classified into three groups, according to the type of histological lesions present in their gut tissues: focal (n = 447), multifocal (n = 59), and diffuse (n = 60). The ELISA based on the detection of ABCA13 was successfully validated showing good discriminatory power between animals with focal lesions and control animals (sensitivity 82.99% and specificity 80.43%). Conversely, the MMP8-based ELISA showed a poor discriminatory power between the different histological groups and non-infected controls. The ABCA13-based ELISA showed a higher diagnostic value (0.822) than the IDEXX ELISA (0.517), the fecal bacterial isolation (0.523) and the real-time PCR (0.531) for the detection of animals with focal lesions. Overall, our results indicate that this ABCA13 ELISA greatly improves the identification of subclinically infected animals with focal lesions that are undetectable using current diagnostic methods, SI, This study was part of the I+D+i project (RTI2018- 094192-R-C22) and was funded by the Spanish MCIN/AEI/10.13039/501100011033/ Ministry of Science, Innovation and the European Regional Development Funds (FEDER Una manera de hacer Europa) and by the Gobierno del Principado de Asturias, Regional funds PCTI 2021– 2023 (GRUPIN: IDI2021-000102) co-funded by FEDER. We acknowledge the National Institute for Agricultural Research (INIA) for the scholarships of CB-V (Ayuda CPD2016-0142 financiada por MCIN/AEI/ 10.13039/501100011033 y FSE El FSE invierte en tu future) and MC

Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/)., [EN] Glaesserella parasuis is the etiological agent of Glässer’s disease (GD), one of the most important diseases afflicting pigs in the nursery phase. We analyzed the genetic and immunological properties of the TbpB protein naturally expressed by 27 different clinical isolates of G. parasuis that were typed as serovar 7 and isolated from pigs suffering from GD. All the strains were classified as virulent by LS-PCR. The phylogenetic analyses demonstrated high similarity within the amino acid sequence of TbpB from 24 clinical strains all belonging to cluster III of TbpB, as does the protective antigen TbpBY167A. Three G. parasuis isolates expressed cluster I TbpBs, indicating antigenic diversity within the SV7 group of G. parasuis. The antigenic analysis demonstrated the presence of common epitopes on all variants of the TbpB protein, which could be recognized by an in vitro analysis using pig IgG induced by a TbpBY167A-based vaccine. The proof of concept of the complete cross-protection between clusters I and III was performed in SPF pigs immunized with the TbpBY167A-based vaccine (cluster III) and challenged with G. parasuis SV7, strains LM 360.18 (cluster I). Additionally, pigs immunized with a whole-cell inactivated vaccine based on G. parasuis SV5 (Nagasaki strain) did not survive the challenge performed with SV7 (strain 360.18), demonstrating the absence of cross-protection between these two serovars. Based on these results, we propose that a properly formulated TbpBY167A-based vaccine may elicit a protective antibody response against all strains of G. parasuis SV7, despite TbpB antigenic diversity, and this might be extrapolated to other serovars. This result highlights the promising use of the TbpBY167A antigen in a future commercial vaccine for GD prevention., SI, This research was funded by Secretaria de Desenvolvimento Econômico Ciência e Tecnologia do Rio Grande do Sul (SDECT, Grant 328–2500/14-0). S.R.P. was supported by Foundation of University of Passo Fundo Master fellowship. S.C. was supported by University of Calgary Eyes High Doctoral Recruitment Award., We thank AFK Imunotech for the generous gift of pigs for this study