Immune response of gilthead seabream (Sparus aurata) after experimental infection with lymphocystis disease virus (LCDV-Sa)

Lymphocystis disease (LCD) is caused by the lymphocystis disease virus (LCDV) (family Iridoviridae), affecting more than 150 fish species from both marine and freshwater environments. A few studies have been focused on the immune defensive mechanisms of fish against LCDV, but only one was conducted during a natural LCD outbreak in gilthead seabream, which is one of the most important cultured fish species in the Mediterranean and the European Atlantic coasts. The aim of this study was the analysis of 23 genes related to the immune response in gilthead seabream specimens after experimental infection with LCDV-Sa using real-time PCR (qRT-PCR) in samples of head kidney and intestine at 1, 3, and 8 dpi. To study the progression of LCDV-Sa infection in gilthead seabreams, the number of viral DNA copies and the expression of mcp were determined in samples of caudal fin, head kidney and intestine. LCDV-Sa was detected by qPCR in all the samples from inoculated fish analysed, whereas no amplification was obtained in samples from the control group. Regarding the gene expression following LCDV-Sa infection, a total of 22 of the 23 genes studied were differentially expressed in head kidney or intestine samples at some time points analysed. Different gene expression profiles were obtained between the organs studied, detecting 18 differentially expressed genes (DEGs) in head kidney samples, four of them exclusively up- or down-regulated (nccrp1, il10, mhcII, and tnfα genes), and 5 genes with a significant change in the expression tendency from 1 to 8 dpi (irf3, isg15, il10, ck10, and c3). In the intestine, 18 DEGs were also detected (14 shared with head kidney), being mx1, casp1, ck3 and tlr9 genes exclusively detected in these samples, and mx1, mx3, irf9 and ighm differentially regulated over time. The results obtained allow us to understand which genes are essential for host-pathogen interactions and could be used as molecular markers for vaccine efficacy evaluation.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech Proyecto de Excelencia Junta de Andalucía Ref. P12-RNM-226

Evaluation of gilthead seabream (Sparus aurata) immune response after LCDV DNA vaccination

A DNA vaccine against Lymphocystis Disease Virus (LCDV) was developed and its protective efficacy in gilthead seabream (Sparus aurata) has been established. The aim of the present study is the evaluation of immune-related gene expression after vaccination to identify which genes could be relevant to control the viral infection. The vaccine was administered intramuscularly to gilthead seabream specimens (100 g weight) at 10 µg/fish dose. In addition, two control groups, injected with the empty plasmid at the same dose or PBS, were established to evaluate non-specific immune response and basal response of fish, respectively. In this study 23 genes related to the immune response (tlr5, tlr9, ifnI, irf1, irf3, irf9, pkr, mx1, mx2, mx3, isg15, tnfα, casp1, il1β, il6, il10, ck3, ck10, c3, nccrp1, mhcII, tcrβ, and ighm) and 2 reference genes (ef1α and actβ) were analysed using real-time PCR (RT-qPCR) in samples of head kidney and intestine at 1, 3, and 8 d post-vaccination. DNA-vaccination of gilthead seabream induced the differential expression of 9 genes in head kidney and 15 genes in intestine samples. Through the course of the experiment, 9 of those genes reached high level of up-regulation comparing to control groups. These genes were related to IFN type I pathway (irf9 and mx3, in head kidney), inflammation (il1β, il6, tnfα, ck10, c3 and nccrp-1, in both organs analysed), and adaptive immune response (mhcII, in intestine). Conclusion: The results obtained allow us to understand which genes could be responsible for the protection against LCDV infection conferred by the DNA vaccine in gilthead seabream. Inflammation is the biological process mainly triggered as a systemic response in vaccinated fish. Different gene expression profiles have been observed in each organ, which may indicate specialized roles relative to immune defensive mechanisms.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

FIRST REPORT OF BURROWING PARROT (CYANOLISEUS PATAGONUS) NESTING IN TREE CAVITIES

Abstract ∙ The Burrowing Parrot Cyanoliseus patagonus is known to breed in burrows mostly on cliffs and ravines in arid or semi‐arid regions of Argentina and Chile. However, during a tree cavity monitoring project we confirmed at least two active nests in tree cavities. Cavity entrances were located between 3.1 and 5.3 m above the ground in live caldén (Prosopis caldenia) trees, Parque Luro, province of La Pampa, Argentina. One nest failed while the other one successfully produced three fledglings. The absence of cliffs and scarcity of ravines in the region, and the presence of a nesting colony of the Blue‐crowned Parakeet (Thectocercus acuticaudatus) in the site may have promoted the adoption of this new nesting substrate for the species. Resumen ∙ Nidificación inusual del Loro Barranquero (Cyanoliseus patagonus) en cavidades naturales de árboles El Loro Barranquero (Cyanoliseus patagonus) nidifica mayoritariamente en cavidades en barrancos y acantilados en regiones áridas y semiáridas de Argentina y Chile. Sin embargo, durante un proyecto de monitoreo de cavidades en árboles, confirmamos al menos dos nidos activos de Loro Barranquero. La entrada de las cavidades estuvo localizada entre 3.1 y 5.3 m de altura en árboles vivos de caldén (Prosopis caldenia), en Parque Luro, provincia de La Pampa, Argentina. Un nido fracasó mientras que el otro tuvo éxito produciendo tres volantones. La escasez de barrancos y la presencia de una colonia de nidificación de Calancate Común (Thectocercus acuticaudatus) en el sitio, podrían haber favorecido la adopción de este nuevo sustrato de nidificación para la especie

Quantifying the available capacity and resource needs for provision of CAR-T therapies in the National Health Service in Spain: a survey-based study

Leukaemia; Organisation of health services; TherapeuticsLeucemia; Organización de servicios de salud; TerapéuticaLeucèmia; Organització dels serveis sanitaris; TerapèuticaObjectives To estimate the readiness of Spanish National Health Service (NHS) hospitals to provide chimeric antigen receptor T cell (CAR-T), and to identify and quantify the different resources needed to provide CAR-T considering three scenarios defined by 10, 25 and 50 patients per centre per year. Design Targeted literature review and quantitative study using a questionnaire and telephone interviews. An algorithm was created to determine hospitals’ readiness based on their capacity and capability. All the requirements for quantification were assessed and validated by the steering committee, formed by members of the Spanish Group of Haematopoietic Transplantation and Cell Therapy. A weighting system (from 0 to 1) was established for capability quantification. For resources quantification, a scoring system was established, with 0 points representing the minimum and 3 points the maximum of additional resources that a hospital indicated necessary. Setting 40 Spanish hospital centres that perform allogeneic haematopoietic stem cell transplantation were invited to complete the questionnaire for capacity quantification, 28 of which provided valid responses. Nine hospitals participated in the interviews for resource quantification, eight of which had previously been designated by the Ministry of Health (MoH) to provide CAR-T. Outcome measure Current capacity of NHS Spanish sites to administer CAR-T under different theoretical scenarios with varying numbers of procedures, and the potential healthcare resources that would be needed to realise the theoretical capacity requirements. Results Four hospitals were optimally ready, 17 were somewhat ready and 7 were not ready. The actual extrapolated capacity of the currently designated MoH CAR-T sites would allow treatment of approximately 250 patients per year. Regarding healthcare resource needs, the numbers of haematologists, nurses and beds were the most important limiting factors, and those requiring further growth as patient numbers increased. Conclusions Increasing the number of CAR-T-qualified centres and/or increasing resources in the current designated sites are two potential strategies that should be considered to treat CAR-T-eligible patients in Spain.This study was conducted with a grant from GETH-TC (grant number: N/A)

Evaluation of immune response after LCDV-Sa infection in DNA-vaccinated gilthead seabream

The immune-related gene expression in vaccinated gilthead seabream after Lymphocystis Disease Virus 3 (LCDV-Sa) infection was analysed by using an OpenArray based on TaqMan qPCR. The DNA vaccine used in this study encodes the viral major capsid protein and confers protection against LCDV-Sa infection in juvenile gilthead seabream. Gilthead seabream juveniles were distributed into four experimental groups and intramuscularly injected with the vaccine (vaccinated group), the empty-plasmid (mock-vaccinated group), or PBS (control groups). Thirty days after vaccination, vaccinated and mock-vaccinated fish, as well as one of the control groups, were injected intraperitoneally with LCDV-Sa (106 TCID50/fish). Samples of head-kidney (HK) from 6 fish were individually collected 1 and 3 days post-infection (dpi). The relative expression levels of 49 genes related to the immune response and 4 reference genes were analysed using an OpenArray. Samples from the non-infected control group were used as calibrator. The number of genes differentially expressed (DEG) in HK at 1 dpi was higher in vaccinated fish compared with both mock-vaccinated and non-vaccinated animals. At 3 dpi, most DEG were upregulated, and the differences in their number among groups were minimized. The recombination-activating gene 1 (rag1), a mediator of development of B and T lymphocytes, was the only gene upregulated in HK samples at 1 dpi. This gene was also upregulated in non-vaccinated animals but at 3 dpi. In contrast, early mx induction was observed in non-vaccinated animals (upregulation of mx2 at 1 dpi) in comparison to vaccinated seabreams (upregulation of mx1 and mx2 at 3 dpi). The results that will be discussed could evidence the role of the DNA vaccine as regulator of the primary lymphoid tissues (HK) in gilthead seabream against LCDV-Sa infection, through downregulation of inflammation related-genes, early upregulation of rag1, and a later expression of interferon stimulated genes.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tec

Immune response of vaccinated juvenile gilthead seabream (Sparus aurata) after LCDV-Sa infection

Lymphocystis disease is one of the main viral pathologies affecting cultured gilthead seabream in the Mediterranean area. In our group, a DNA-vaccine has been developed based on the major capsid protein (MCP) of the Lymphocystis Disease Virus 3 (LCDV-Sa). The aim of the present study is the evaluation of immune-related gene expression in vaccinated fish after viral infection to identify immunogenes involved in the vaccine-induced protection. To fulfil this objective an OpenArray® platform has been developed to study 49 genes related to the immune response. Reference and viral genes were also evaluated. Gilthead seabream specimens (5 g mean weight) were distributed into 3 experimental groups, inoculated with the vaccine at 0.1 µg/g fish dose, the empty plasmid at the same dose or PBS. Thirty days post-vaccination, fish were intramuscularly injected with the virus at 106 TCID50/fish dose. Samples of head-kidney, spleen, intestine and caudal fin from 6 fish were individually collected at 1, 2 and 3-days post-injection in all groups. The quantification of viral DNA in fins of fish challenged with LCDV-Sa were carried out by a qPCR assay targeting a viral structural gene (putative myristoylated membrane protein, MMP) alternative to the mcp gene contained in the vaccine. The results obtained showed an increase of genes deregulated within the haematopoietic organs between vaccinated and non-vaccinated fish. However, in the intestine and fin, the results showed the opposite trend. The global effect of fish vaccination was a diminished immune response compared to non-vaccinated fish, being 83 and 99 genes differentially expressed through the experiment, respectively. Moreover, viral replication decreased in groups of fish previously vaccinated. The modulation of the immune response provoked by the vaccination trial seems to control the progression of the disease.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tec

Biomedical point-of-care microanalyzer for potentiometric determination of ammonium ion in plasma and whole blood

Acord transformatiu CRUE-CSICSome inborn errors of metabolism and other diseases can result in increasing blood ammonium (hyperammonemia episodes), which can cause serious neurological complications in patients or even death. Early diagnosis, follow up and treatment are essential to minimize irreversible damages in brain. Currently, adequate analytical instrumentation for the necessary ammonium bedside determination is not available in all health centers but only in clinical laboratories of reference hospitals. We therefore have developed a low cost and portable potentiometric Point-of-Care microanalyzer (POC) to address this problem. It consists of a cyclic olefin copolymer-based microanalyzer, the size of a credit card and working in continuous flow, which integrates microfluidics, a gas-diffusion module and a potentiometric detection system. The analytical features achieved are a linear range from 30 to 1000 μmol L NH , a detection limit of 18 μmol L NH and a required sample volume of 100 μL, which comply with the medical requirements. Plasma and blood samples are analyzed with no significant differences observed between ammonium concentrations obtained with both the proposed microanalyzer and the reference method. This demonstrates the value of the developed POC for bedside clinical applications

Los programas de educación náutica en la educación obligatoria en España: revisión sistemática

En los últimos años, los deportes náuticos están adquiriendo notoriedad en el ámbito educativo, pudiendo demostrar que son una herramienta valiosa para el desarrollo integral de los estudiantes. Sin embargo, no parece existir evidencia científica sobre el desarrollo y efecto de estos en los programas de educación náutica en España. Por ello, el objetivo de este estudio fue la realización de una revisión sistemática de los programas relacionados con la educación náutica en la educación Primaria y Secundaria en España. Además, proponer un meta-programada en la línea de esta temática. A partir de estos objetivos, se realizó la búsqueda en las bases de datos PubMed, Scopus, Dialnet, Web of Science y, posteriormente, en Google Académico y en el buscador de Google, entre 2008 hasta 2023 empleando las siguientes palabras clave: "programa escolar", "educación", "actividades náuticas".  Los resultados de la búsqueda muestran que los principales objetivos de estos programas son el fomento de actividades y deportes náuticos y el conocimiento del propio entorno. En cuanto a los contenidos, destaca el uso de los deportes de vela, y otros deportes como son piragüismo, windsurf, kayak o paddle surf, que empiezan por clases teóricas, para posteriormente, realizar las clases prácticas. En conclusión, se ha podido comprobar que, en la actualidad, se está haciendo un uso deportes náuticos en el ámbito educativo, lo que nos ha llevado a realizar un meta-programa donde se recojan la información más relevante de los objetivos, contenidos, metodología y evaluación

Análisis de la expresión de inmunogenes en doradas vacunadas en respuesta a la infección por LCDV-Sa

El objetivo del presente estudio ha sido evaluar la respuesta inmune frente a la infección por Lymphocystis Disease Virus 3 (LCDV-Sa) en ejemplares de dorada (Sparus aurata) vacunados con un plásmido que codifica la proteína principal de la cápside del virus, con el fin de identificar inmunogenes relacionados con la protección inducida por la vacuna. Se utilizaron juveniles de dorada inyectados intramuscularmente con la vacuna (grupo vacunado) o el plásmido vacío (grupo placebo), así como con PBS (dos grupos). A los 30 d post-vacunación, los peces se inocularon con un aislado de LCDV-Sa (106 TCID50/pez), excepto uno de los grupos originalmente inyectado con PBS que se mantuvo como control no-infectado. Se tomaron muestras de riñón cefálico a los 1 y 3 d post-infección (dpi) (6 peces por tiempo) y se realizó un análisis de la expresión relativa de 49 inmunogenes de dorada utilizando la plataforma OpenArray®, basada en qPCR con sondas TaqMan, usando las muestras del control no-infectado como calibrador. Los resultados mostraron una mayor expresión diferencial de inmunogenes en los peces vacunados en comparación a los peces que recibieron el plásmido vacío o a los no vacunados. En todos los grupos experimentales se observó una sub-regulación génica a 1 dpi, que cambió a sobre-regulación a los 3 dpi. Además, en los peces vacunados se observó la estimulación temprana de la expresión del gen activador de recombinación (rag1) y una sobre-regulación tardía de mx1 y mx2.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech