Histopathological changes in the skin and gut mucus layers of rainbow trout (Oncorhynchus mykiss) challenged with Ichthyophthirius multifiliis inactivated by gamma rays and formalin

Ichthyophthirius multifiliis, a protozoan parasite, is a significant problem for fish farmers and Aquarium fish worldwide. This study aimed to evaluate the immunization of rainbow trout with gamma-irradiated, formalin inactive, and live theronts of I. multifiliis. In this study, fish were exposed to gamma-irradiated, formalin-inactivated, and live I. multifiliis theronts. Then, the histopathological changes in the mucous layers of the skin and intestines were studied after 7 and 14 days of exposure. Although no significant morphological changes were observed in the skin and intestines of the treated fish, the number of skin goblet cells increased significantly in fish treated with formalin-inactivated, gamma-inactivated, and live trophonts on 7 and 14 days. Compared to the negative control group, an increase in epidermal thickness on the skin was observed in fish challenged with formalin-inactivated, gamma-inactivated, and live trophonts. The numbers of mucous cells/total enterocytes in the intestinal epithelium of fish exposed to gamma-irradiated, formalin-inactivated, and trophonts live were higher than in non-infected fish. Moreover, a significant increase was found in the mucous cell numbers of the pyloric fold in treated fish with gamma-irradiated and formalin inactive trophonts at the first and second weeks. The results showed that the gamma-irradiated trophonts and formalin inactive trophonts could be safe for use in rainbow trout against I. multifiliis

Immune modulation by virus-encoded secreted chemokine binding proteins

Chemokines are chemoattractant cytokines that mediate the migration of immune cells to sites of infection which play an important role in innate and adaptive immunity. As an immune evasion strategy, large DNA viruses (herpesviruses and poxviruses) encode soluble chemokine binding proteins that bind chemokines with high affinity, even though they do not show sequence similarity to cellular chemokine receptors. This review summarizes the different secreted viral chemokine binding proteins described to date, with special emphasis on the diverse mechanisms of action they exhibit to interfere with chemokine function and their specific contribution to virus pathogenesis.Spanish Ministry of Economy and Competitiviness (grants SAF2009-07857 and SAF2012-38957)Peer Reviewe

Ključna uloga faktora tumorske nekroze alfa (TNF-α) u kalifornijskih pastrva cijepljenih ozračenim trofontom parazita Ichthyophthirius multifiliis.

In this study, in order to characterize the immune response against Ichthyophthirius multifiliis (I. multifiliis) in the skin, liver, gills and head kidney of immunized rainbow trout, with two types of killed vaccines (γ-irradiation and formalin inactivation of trophont), the gene expression levels of the pro-inflammatory cytokines tumour necrosis factor-α (TNF-α1 and TNF-α2) were evaluated. The vaccinated fish showed significant protection against I. multifiliis 30 days after the second vaccination. We showed that the pro-inflammatory cytokine, TNF-α1, was expressed in rainbow trout after vaccination, not only in the skin but also in the head kidney and liver, whereas TNF-α2 expression was seen in the liver. Also, parasite-related changes in TNF-α1 expression could be detected only in the gills of fish that were exposed to live I. multifiliis trophonts during this experiment. Finally, according to previous reports and the current study, TNF-α1 could be involved in an immune mechanism that can control I. multifiliis infection in vaccinated rainbow trout.Istražen je imunosni odgovor u koži, jetri, škrgama i bubregu kalifornijskih pastrva cijepljenih dvjema vrstama inaktiviranih cjepiva pripravljenih od parazita Ichthyophthirius multifiliis. Jedna je bilo pripravljena ozračivanjem trofonta γ-zrakama, a druga njegovim ubijanjem formalinom. Istražena je razina genske ekspresije proupalnih citokina odnosno faktora tumorske nekroze α (TNF-α1 i TNF-α2). Cijepljene ribe pokazivale su značajnu zaštitu protiv I. multifiliis 30 dana nakon drugog cijepljenja. Pokazalo se da je proupalni citokin TNF-α1 bio izražen u pastrva nakon cijepljenja ne samo u koži već i u bubregu i jetri, dok je ekspresija TNF-α2 bila dokazana samo u jetri. U ovom je pokusu također ustanovljeno da se promjene u ekspresiji TNF-α1 mogu dokazati samo u škrgama riba izloženima živim trofontima I. multifiliis. Na osnovi prijašnjih izvješća i ovog istraživanja može se zaključiti da bi TNF-α1 mogao biti upleten u imunosne mehanizme za kontrolu invazije vrstom I. multifiliis u cijepljenih kalifornijskih pastrva

Sistema de sensorización para el control de clima en cultivos

El presente proyecto pretende ser un acercamiento al mundo de Internet of Things a través de una serie de experimentos que van mutando en su estructura, que no en su funcionalidad, a lo largo del desarrollo del mismo. El objetivo principal ha sido recoger información del entorno más próximo mediante sensores y actuar sobre el entorno a través de actuadores conectados a diferentes elementos eléctricos o electrónicos que puedan ser accionados y puestos en marcha de manera remota. Lo que ofrece Internet of Things, y esta aplicación en concreto, es una especie de ubicuidad al poder situar estos Objetos en cualquier entorno con conectividadWi-Fi y alimentación eléctrica. Esto permite que dichos espacios adquieran cierta inteligencia al tener recuerdo de las situaciones pasadas de manera que podremos actuar sobre ellos con mayor conocimiento. El presente trabajo ha sido ideado para conseguir datos relevantes de un cultivomediante su sensorización y tener la posibilidad de actuar sobre él de manera remota accionando elementos eléctricos o electrónicos. Algo a resaltar de este trabajo, es la polivalencia que presenta el sistema al permitir ser adaptado e implementado a otros entornos con relativa facilidad. Mediante cambios en elModelo de Negocio puede ser utilizado incluso en áreas como la domótica.Grado en Ingeniería Informátic

Caracterización molecular y funcional de la proteína de unión a quimioquinas E163 de ECTV

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 29-10-2015Esta tesis doctoral ha sido realizada en el Centro de Biología Molecular Severo Ochoa con la financiación de una de una ayuda Predoctoral de Formación de Personal Investigador (referencia BES-2010-030612) asociada al proyecto de investigación SAF2009-0785

Molecular Methods for Identification of Acinetobacter Species by Partial Sequencing of the rpoB and 16S rRNA Genes.

BACKGROUND Acinetobacter spp. is a diverse group of Gram-negative bacteria which are ubiquitous in soil and water, and an important cause of nosocomial infections. The purpose of this study was to identify a collection of Acinetobacter spp. clinical isolates accurately and to investigate their antibiotic susceptibility patterns. MATERIALS AND METHODS A total of 197 non-duplicate clinical isolates of Acinetobacter spp. isolates identified using conventional biochemical tests. The molecular technique of PCR-RFLP and sequence analysis of rpoB and 16S rRNA genes was applied for species identification. Antimicrobial susceptibility test was performed with a disk diffusion assay. RESULTS Based on 16S rRNA and rpoB genes analysis separately, most of clinical isolates can be identified with high bootstrap values. However, the identity of the isolate 555T was uncertain due to high similarity of A. grimontii and A. junii. Identification by concatenation of 16S rRNA and rpoB confirmed the identity of clinical isolates of Acenitobacer to species level confidently. Accordingly, the isolate 555T assigned as A. grimontii due to 100% similarity to A. grimontii. Moreover, this isolate showed 98.64% to A. junii. Besides, the identity of the isolates 218T and 364T was confirmed as Genomic species 3 and A. calcoaceticus respectively. So, the majority of Acinetobacter spp. isolates, were identified as: A. baumannii (131 isolates, 66%), A. calcoaceticus (9 isolates, 4.5%), and A. genomosp 16 (8 isolates, 4%). The rest of identified species showed the lower frequencies. In susceptibility test, 105 isolates (53%), presented high antibiotic resistance of 90% to ceftriaxone, piperacillin, piperacillin tazobactam, amikacin, and 81% to ciprofloxacin. CONCLUSION Sequence analysis of the 16S rRNA and rpoB spacer simultaneously was able to do identification of Acinetobacter spp. to species level. A.baumannii was identified as the most prevalent species with high antibiotic resistance. Other species showed lower frequencies ranged from 4 to 9 strains

Varijacije ukupne razine sijalične kiseline u bubrezima, jetri i slezeni imunizirane kalifornijske pastrve (Oncorhynchus mykiss)

The aim of this study is to evaluate the effects of irradiated trophonts of Ichthyophthirius multifiliis and alginate/calcium phosphate nanoparticles on total sialic acid levels in immunized rainbow trout kidney, liver and spleen tissues. The 15 fish tissues of the treated and control groups were sampled at 30 days following the first immunization. Tissue samples were homogenized and centrifuged. The total sialic acid levels of the tissue samples were determined using a commercially available Quantitation Kit. Significant increases in kidney, spleen and liver TSA levels were determined in the treated groups in comparison to the controls. The immunosupportive effects of sialic acid were illustrated by the elevation in the amount of TSA in the kidneys, liver and spleen of the immunized fish. Also, the significant increase in the TSA levels in the rainbow trout treated with calcium phosphate nanoparticles could be attributed to the Ca-binding capacity of the glycerol side chain of the sialic acid. Finally, the results of this study showed that sialic acid may be a mediator in the development of long-circulating nanocarriers to advance the field of vaccine delivery in fish.Cilj ovoga istraživanja bio je procijeniti učinak ozračenog trofonta Ichthyophthirius multifiliis i nanočestica alginata / kalcijeva fosfata na ukupnu razinu sijalične kiseline u bubrezima, jetri i slezeni imunizirane kalifonijske pastrve. Uzeti su uzorci 15 riba iz pokusne i kontrolne skupine 30 dana nakon imunizacije. Uzorci tkiva su homogenizirani i centrifugirani. Ukupna razina sijalične kiseline u uzorcima tkiva utvrđena je komercijalnim kitom za kvantifikaciju. U pokusnim skupinama u usporedbi s kontrolnom utvrđen je statistički znakovit porast ukupne razine sijalične kiseline u bubregu, jetri i slezeni. Imunopotporni učinci sijalične kiseline prikazani su porastom ukupne razine sijalične kiseline u bubrezima, jetri i slezeni imunizirane ribe. Također, statistički znakovit porast ukupne razine sijalične kiseline u kalifonijske pastrve kojoj su davane nanočestice kalcijeva fosfata, mogao bi se povezati sa sposobnošću postranog lanca glicerola sijalične kiseline za vezanje kalcija. Rezultati ovoga istraživanja pokazali su da sijalična kiselina može posredovati u razvoju dugocirkulirajućih nanonositelja kako bi se pospješila imunizaciju riba

Rapid Detection of Rifampicin- and Isoniazid-Resistant Mycobacterium tuberculosis using TaqMan Allelic Discrimination

Objectives: Multidrug-resistant tuberculosis (MDR-TB) is a global problem that many countries are challenged with. Rapid and accurate detection of MDR-TB is critical for appropriate treatment and controlling of TB. The aim of the present study was to evaluate the TaqMan allelic discrimination without minor groove binder (MGB) as a rapid, efficient, and low-cost method for detection of drug resistant strains of Mycobacterium tuberculosis. Methods: A total of 112 M. tuberculosis isolates from cases with diagnosed TB were subjected to drug susceptibility testing (DST), using the proportion method. Resistant isolates were tested for characterization of mutations in the rpoB and KatG genes by TaqMan genotyping. Results: Of 112 M. tuberculosis isolates for which DST was performed, three, one, and two isolates were MDR, rifampin (RIF) resistant, and isoniazid (INH) resistant, respectively. According to the threshold cycle (Ct) and curve pattern of mutants, TaqMan probes detect all of the mutations in the analyzed genes (katG 315, AGC�ACC, rpoB 531, TCG�TTG, and rpoB 531, TCG�TGG). Conclusion: The present study suggests that drug-resistant strains of M. tuberculosis can be detected by pattern's curve or Ct with TaqMan probes without MGB in real-time polymerase chain reaction (PCR). © 2016

Chemokines cooperate with TNF to provide protective anti-viral immunity and to enhance inflammation

The role of cytokines and chemokines in anti-viral defense has been demonstrated, but their relative contribution to protective anti-viral responses in vivo is not fully understood. Cytokine response modifier D (CrmD) is a secreted receptor for TNF and lymphotoxin containing the smallpox virus-encoded chemokine receptor (SECRET) domain and is expressed by ectromelia virus, the causative agent of the smallpox-like disease mousepox. Here we show that CrmD is an essential virulence factor that controls natural killer cell activation and allows progression of fatal mousepox, and demonstrate that both SECRET and TNF binding domains are required for full CrmD activity. Vaccination with recombinant CrmD protects animals from lethal mousepox. These results indicate that a specific set of chemokines enhance the inflammatory and protective anti-viral responses mediated by TNF and lymphotoxin, and illustrate how viruses optimize anti-TNF strategies with the addition of a chemokine binding domain as soluble decoy receptors.We thank Javier Salguero for help with animal experimentation and immunohistochemistry, Rocío Martín and Carolina Sánchez for technical assistance and Daniel Rubio for discussions on the project. This work was funded by Grants from the Spanish Ministry of Economy and Competitiviness and European Union (European Regional Development’s Funds, FEDER) (grant SAF2015-67485-R), and the Wellcome Trust (grant 051087/Z97/Z). M.B.R.-A. and A. Alejo were recipients of a Ramón y Cajal Contract from the Spanish Ministry of Science and Innovation