A scanning electron microscopy study of Anisakis physeteris molecularly identified: from third stage larvae from fish to fourth stage larvae obtained in vitro

The development of the fourth larval stage (L4) of Anisakis physeteris was studied using scanning electron microscopy (SEM), comparing it with third larval stage (L3) recently obtained from the host fish, blue whiting (Micromesistius poutassou), from the western Mediterranean Sea (east coast of Spain, zone FAO 37.1.1). After molting to L4, samples of the parasite were examined at different times in order to observe their development. Following collection of the L4, a small portion was taken from the middle of the larva for molecular identification, confirming in all cases that it was A. physeteris. The anterior and posterior sections of the larvae were prepared for morphological study by SEM. The development of a row of denticles on each of the three prominent lips, almost reaching the buccal commisures, was observed in the L4. Pores of unknown function were found in the upper external part of each lip. Clearly developed cephalic papillae, amphids, and deirids were also observed in L4, while, although present in L3, these were beneath the cuticle. Phasmids were detected in L4 but not in L3. The L4 tail finished in a conical lobe with a blunt point, absent in L3. In the oldest L4, some preanal papillae were observed beneath the cuticle in males, while, in females, the vulva could be seen by light microscopy, apparently still covered by the cuticle.This work has been funded by the Agencia Estatal de Investigación (Spanish State Research Agency) and European Regional Development Fund (ERDF), grant number CGL2013-47725-P

Results of a computer application for self-evaluation of the practical classes of Parasitology in the Pharmacy Degree

Se ha desarrollado una aplicación para los alumnos de “Parasitología” (2º curso de Farmacia) que contiene información e imágenes de parásitos obtenidas de las mismas preparaciones que observan en las clases prácticas. Con ella pueden repasar y autoevaluar los conocimientos adquiridos. Para evaluar el proyecto, se ha permitido el acceso ilimitado a la aplicación a 74 alumnos, durante los 9 días en que realizaron sus prácticas. Las calificaciones obtenidas han sido comparadas con un grupo control de 75 alumnos que realizó las prácticas el curso anterior en fechas coincidentes con las del grupo de este trabajo y con los mismos profesores. La comparación de las calificaciones arroja un incremento medio de 0,7 puntos (sobre 10). Estudiando estos datos según rangos de puntuación, se observa que el grupo experimental ha disminuido el porcentaje de suspensos (2,50%), incrementándose 9,95% los alumnos con calificación ≥9,0 (sobre 10). Estos alumnos han contestado una encuesta anónima en la que nos dan su opinión acerca de la usabilidad, organización, diseño y calidad de la aplicación, suficiencia de la información aportada, utilidad de la autoevaluación, mejora del aprendizaje y resultados obtenidos en la autoevalución de la aplicación, así como su opinión sobre posibles mejoras de la misma, valorando entre 1 (pésimo) y 5 (óptimo). La encuesta, respondida por 50 alumnos de 74, arrojó una media global de 4,28 sobre 5. El alumnado expresó su satisfacción por contar con esta herramienta de apoyo a las prácticas.A computer application for students in "Parasitology (2nd year of Pharmacy) that contains information and pictures of the parasites obtained from the same preparations that observed in practical classes has been developed. With this computer tool, the students can review and self-evaluation their learning. For the evaluation of the project, a 74 students group was allowed unlimited access to the computer application during the 9 days that did their practices. The scores obtained have been compared with a control group of 75 students who did the practice the previous year on dates coinciding with the group of this study and with the same teachers. The comparison of the ratings shows an average increase of 0.7 points (on 10). Studying these data according to score ranges, it appears that the experimental group decreased the failure rate (2.50%), increasing 9.95% rating students with ≥ 9.0 (on 10). These students have answered an anonymous survey in which they give their opinion about the usability, organization, design and quality of implementation, fitness of provided information, the utility of selfevaluation, improvement of learning and evaluation their results in the implementation as well as their views on possible improvements to it, from 1 (very poor) to 5 (excellent). The survey was answered by 50 students from 74, giving an overall average of 4.28 points on 5. The students expressed their satisfaction with this computer tool available to support practices.Este trabajo ha sido realizado gracias al proyecto PID 08-50, subvencionado por la Unidad de Innovación Docente de la Universidad de Granada

Elaboración de un manual de prácticas para su utilización en asignaturas impartidas por el Departamento de Parasitología

Esta propuesta se presenta como segunda fase del proyecto 20-115, desarrollado durante el periodo 2020-2022 (Plan FIDO UGR). Durante dicho proyecto se han digitalizado 1500 imágenes de parásitos que engloban multitud de especies, incluyendo: protozoos, trematodos, cestodos, nematodos y artrópodos, así como 12000 fotografías necesarias para obtener otras tantas imágenes por la técnica del apilado. Con el ingente material producido se ha elaborado un protocolo de prácticas común para ser utilizado por el alumnado que realice sus prácticas en asignaturas impartidas por el Departamento de Parasitología.Vicerrectorado de Calidad, Innovación Docente y Estudios de Grado-Universidad de Granad

Cultivo, en condiciones experimentales, de Bovicola Caprae Gurlt y Bovicola Limbata Gervais y sus aplicaciones a estudios taxonómicos

Se ha realizado un estudio morfologico y biologico de los malófago bovicola caprae gurlt 1843 y bovicola limbata gervais 1844 parásitos de capra hircus l. estudiándose la supervivencia en estado de ayuno o alimentados con pelo descamaciones dérmicas levadura de cerveza glutámico medio m.e.m. y/o medio grace a 35+-1 5 c y 75+-5% de humedad relativa y/o a 20+-10 c y 55+-10% de h.r. igualmente se ha estudiado el ciclo biológico de ambas especies teniendo en cuenta datos tales como periodo de preovoposicion de embrionamiento duración del ciclo de huevo a huevo descendencia obtenida etc. siendo la 1a vez que se cierra en el laboratorio el ciclo biológico de bovicola caprae gurlt 1843. así mismo se ha estudiado la partenogénesis como vía de reproducción alternativa la existencia de neotenia o cruce entre estas especiesTesis Univ. Granada. Departamento de Parasitología. Leída en 198

Molecular Epidemiology of Anisakis spp. in Wedge Sole, Dicologlossa cuneata (Moreau, 1881), from Fishmarkets in Granada (Southern Spain), Caught in Two Adjacent NE and CE Atlantic Areas

The presence of third stage larvae (L3) of Anisakis spp. in wedge sole, Dicologlossa cuneata (Moreau, 1881), purchased in fishmarkets in the city of Granada (Andalusia, southern Spain) was assessed. The wedge sole were caught in two FAO zones: area 27.IXa NE Atlantic (SW Spain coast) and area 34.1.11 CE Atlantic (NW Morocco coast). Only Anisakis larvae, type I, were detected in the largest fish (>20 cm) from the CE Atlantic. These were molecularly identified as A. simplex s.s. The prevalence (P) of Anisakis in this area was 12.5% and the mean intensity (MI) was 1. The presence of Hysterothylacium spp. larvae was also detected in the fish from both areas, with the prevalence being approximately double in the CE Atlantic area (12.5 vs. 5.7). A comparison between the Anisakisinfected and non-infected fish from this area showed that the former were significantly longer than the latter (p < 0.01). These results show that Anisakis parasitization of wedge sole sold in the markets of the city of Granada is of low prevalence and intensity (P = 4.5, MI = 1), especially in those from area 27.IXa (P = 0), indicating that the risk of human infection is low, particularly as this fish is traditionally prepared by deep-frying in oil in Andalusia (southern Spain)

Anisakis Infection in the Spotted Flounder Citharus linguatula (Pleuronectiformes: Citharidae) Caught in the Gulf of Cadiz (Area FAO 27-ICES IXa) Appears to Negatively Affect Fish Growth

Spotted flounder (Citharus linguatula L.) caught in the Gulf of Cadiz (area FAO 27 ICES IXa) were examined for Anisakis larvae and to assess the possible risk of anisakiasis in humans through consumption of this fish. Larvae of the genera Anisakis and Hysterothylacium were identified in the analysis of 128 purchased fish specimens. All Anisakis larvae corresponded to type I. Molecular analysis showed the presence of A. pegreffii, A. simplex s.s., and recombinant genotype between the two. The prevalence of Anisakis was 9.4% with a mean intensity of 1.42, while for Hysterothylacium the values were 12.5% and 1.06. The length and weight of the fish, but not Fulton's condition factor, varied significantly between infected and uninfected fish. The prevalence of Anisakis increased with fish length, with no fish parasitized with Anisakis measuring less than 15.5 cm (2-2.5 years old), which is probably related to the reported dietary change of these fish at around 2 years of age. Fish not parasitized with any of these nematodes showed positive allometric growth, while those parasitized only with Anisakis showed negative allometric growth. When comparing both groups including only fish >= 15.5 cm (the smallest size of Anisakis-infected fish), the difference is shown to be statistically significant (p = 0.01), suggesting that Anisakis infection of spotted flounder negatively affects fish growth even when parasite intensity is low, which may have important economic repercussions. Finally, the low prevalence and, above all, intensity of Anisakis in these fish, as well as the habit of consuming this fish fried in oil in our geographical area, means that the risk of acquiring anisakiasis through consumption of this fish is low.Research Group BIO-243 Junta de Andalucía (Spain

Anisakis infection in anchovies (Engraulis encrasicolus) from Iberian waters, southwestern Europe: Post-mortem larval migration

The Anisakis larvae presence in fish for human consumption is a health risk that needs to be monitored. The anchovy is a fish that is highly appreciated by consumers and that can harbour Anisakis. It is thus necessary to periodically evaluate the presence of anisakid larvae in them. So, anchovies from Iberian Peninsula coasts were analysed. Fish examination for macroscopic nematodes showed L3s of both Anisakis type I and Hysterothylacium aduncum. The Anisakis prevalence varies with the catching area and the fish size. The muscle prevalence was 7.45% (mean intensity 1.75; range 1–5). Molecular analysis showed 110 A. simplex s.s. (17 in muscle), 22 A. pegreffii (3) and 7 hybrid genotype individuals (1). Considering that most of the Iberian Peninsula coasts are a sympatry area between these two Anisakis species, it has been observed that A. simplex s.s./A. pegreffii ratio increases from south to north in a clockwise direction. Also, 19 larvae were detected on the fish surface from the Bay of Biscay, indicating the ability of these larvae to migrate after the fish death. The A. simplex s.s./A. pegreffii larvae proportion found on the anchovy surface is similar to the found in viscera and lower than in muscle, suggesting that most of the larvae migrating to the surface must have come from the visceral package. This confirms the importance of removing fish viscera immediately after capture, for those fish species where this is possible. As both species cause anisakiasis/anisakidosis, these data show a real risk to human health, especially in dishes highly prized in Mediterranean countries prepared with raw or semi-raw anchovies.BIO-243 research group of the Junta de Andalucía (Spain)Open access charge: Universidad de Granada / CBUA

Protocolo de Prácticas de Parasitología General 2022-2023

Contiene información sobre las fases y estructuras identificativas de los parásitos, objetivos de cada práctica, metodología, aumento aconsejable para su búsqueda y una serie de cuestiones que los alumnos deben contestar individualmente antes de entregar los protocolos para su corrección por parte del profesor

DNA methylation of miR-7 is a mechanism involved in platinum response through MAFG overexpression in cancer cells

One of the major limitations associated with platinum use is the resistance that almost invariably develops in different tumor types. In the current study, we sought to identify epigenetically regulated microRNAs as novel biomarkers of platinum resistance in lung and ovarian cancers, the ones with highest ratios of associated chemo-resistance. [Methods]: We combined transcriptomic data from microRNA and mRNA under the influence of an epigenetic reactivation treatment in a panel of four paired cisplatin -sensitive and -resistant cell lines, followed by real-time expression and epigenetic validations for accurate candidate selection in 19 human cancer cell lines. To identify specific candidate genes under miRNA regulation, we assembled “in silico” miRNAs and mRNAs sequences by using ten different algorithms followed by qRT-PCR validation. Functional assays of site-directed mutagenesis and luciferase activity, miRNAs precursor overexpression, silencing by antago-miR and cell viability were performed to confirm their specificity in gene regulation. Results were further explored in 187 primary samples obtained from ovarian tumors and controls. [Results]: We identified 4 candidates, miR-7, miR-132, miR-335 and miR-148a, which deregulation seems to be a common event in the development of resistance to cisplatin in both tumor types. miR-7 presented specific methylation in resistant cell lines, and was associated with poorer prognosis in ovarian cancer patients. Our experimental results strongly support the direct regulation of MAFG through miR-7 and their involvement in the development of CDDP resistance in human tumor cells. [Conclusion]: The basal methylation status of miR-7 before treatment may be a potential clinical epigenetic biomarker, predictor of the chemotherapy outcome to CDDP in ovarian cancer patients. To the best of our knowledge, this is the first report linking the regulation of MAFG by miRNA-7 and its role in chemotherapy response to CDDP. Furthermore, this data highlights the possible role of MAFG as a novel therapeutic target for platinum resistant tumors.FIS (ISCIII) and FEDER/FSE funds (PI12/00386, PI12/01463, PI14/01495, PI15/00186, and FEDER/FSE, Una manera de hacer Europa support the study and contracts of I.I.C. and O.V. PTA2012/7141-I funds support OP contract and I.I.C. was financed by the ''Miguel Servet'' program (CP 14/00008). The authors gratefully acknowledge the Colombian Department for Science, Technology and Innovation (COLCIENCIAS), Cod.568-2012, for partially funding this work to J.S.Peer reviewe

Imagen digital aplicada a la docencia teórica y práctica en asignaturas de Parasitología

Este proyecto permitirá obtener un repositorio digital de imágenes de parásitos que serán clasificadas y puestas a disposición de profesionales para su uso en el diseño de clases teóricas y prácticas, y para el estudio de asignaturas relacionadas por el alumnado. Para su desarrollo nos beneficiaremos de las numerosas preparaciones parasitológicas disponibles en el Dpto. de Parasitología de la UGR y de diferentes dispositivos de microscopía óptica que serán de gran utilidad en el desarrollo del proyecto. Las muestras de los parásitos, una vez clasificadas siguiendo criterios diferenciales, será utilizadas para la captación de imágenes digitales de alta resolución que serán incluidas en una base de datos donde se registrarán las principales estructuras morfológicas diferenciales que pudiesen ayudar a cualquier persona interesada en la identificación de dichas especies. La base de datos recopilada se empleará para la elaboración de una guía que sirva como soporte para la realización de las prácticas de aquellas asignaturas con contenidos afines a los referidos. Dicha guía contendrá imágenes identificadas y detalladas (p.e. distintos caracteres diferenciales) de las especies parásitas más relevantes desde los puntos de vista clínico y alimentario. Con todo ello el alumnado y/o profesionales interesados tendrán la opción de estudiarlas sobre el texto impreso y recurrir al repositorio digital para su visualización con mayor detalle. Por ello, se contempla la posibilidad de adjuntar al texto un DVD con las imágenes en mayor calidad. Las imágenes serán distribuidas con licencia Creative commons para su libre distribución y uso no comercial. En el diseño del proyecto se considerará una segunda fase, posterior a los dos años de la presente convocatoria, que incluiría la creación de una página web interactiva (o actualización de los contenidos digitales ya existentes) con las imágenes recopiladas. Esta fase garantizaría la continuidad del proyecto dado que la mejora de los contenidos interactivos siempre admite actualizaciones. En este sentido se contempla que: además de nuevas imágenes se incluirían protocolos de diagnóstico parasitológico, protocolos de manejo y calibración de útiles para dicho diagnóstico como puede ser el microscopio óptico, e imágenes de nuevos especímenes o de mayor calidad, entre otros.Universidad de Granada Unidad de Calidad, Innovación Docente y Prospectiva Plan FIDO 20-22 Código 20-11