02.08. Área de Salud VIII Mar Menor

Molecular epidemiology of Dientamoeba fragilis coinfections with Blastocystis sp. and Giardia duodenalis in symptomatic patients from Valencia, Spain

2026-08-03T10:47:20Z

Molecular epidemiology of Dientamoeba fragilis coinfections with Blastocystis sp. and Giardia duodenalis in symptomatic patients from Valencia, Spain García-Hita, Marta; Ferriz, Jorge; Cifre, Susana; Tapia-Veloz, Gabriela; Molina, Araceli; Carmena, David; Trelis, María PURPOSE: Dientamoeba fragilis is a cosmopolitan intestinal protist with an insufficiently understood life cycle and pathogenicity. Diagnosis by light microscopy is challenging, leading to significant underdiagnosis. Two genotypes (1 and 2) have been described, with genotype 1 being the most frequently detected worldwide. As no molecularly characterized isolates from Spain have been reported, we performed a molecular characterization and evaluated sociodemographic factors of D. fragilis in patients coinfected with Blastocystis sp. and/or Giardia duodenalis. METHODS: Using a multiplex qPCR assay (Allplex? GI-Parasite Assay, Seegene), we analyzed 354 stool samples previously identified as positive for Blastocystis sp. (n = 276), G. duodenalis (n = 63), or both (n = 15) by light microscopy. Eligible positive samples were selected for conventional PCR targeting the SSU rRNA gene and subsequent Sanger sequencing. RESULTS: D. fragilis was detected in 34.5% of samples by qPCR; 32.3% of Blastocystis-positive, 44.9% of Giardia-positive and 46.7% of coinfected samples. We successfully sequenced 22 isolates, all corresponding to genotype 1, identifying two distinct intra-genotypic sequence variants. Younger age was positively associated with D. fragilis, with individuals coinfected with Blastocystis sp. being markedly younger than those monoinfected. Patients with G. duodenalis and D. fragilis were mainly under 15 years of age. CONCLUSION: This study provides the first genotyping of D. fragilis in Spain, demonstrating the circulation of two genotype 1 variants. Findings highlight the underdiagnosis of D. fragilis and its high prevalence in young patients with other fecal-orally transmitted protists. Evaluating coinfections is crucial to understanding parasite relationships and should be considered for clinical management, especially when symptoms persist.

Gene editing of the GJB2 locus in porcine embryos using CRISPR/Cas9 and cytosine base editors: toward a model of congenital deafness

2026-08-03T10:48:13Z

Gene editing of the GJB2 locus in porcine embryos using CRISPR/Cas9 and cytosine base editors: toward a model of congenital deafness Pineiro-Silva, Celia; Bermejo-Álvarez, Pablo; García-Purrinos, Francisco-José; Gadea-Mateos, Joaquín Mutations in the GJB2 gene, which encodes Connexin 26 (Cx26), are responsible for the majority of cases of non-syndromic congenital hearing loss in humans. While murine GJB2 knockout models have provided mechanistic insight, anatomical and physiological differences limit their translational relevance. Pigs represent a valuable large-animal model because their auditory anatomy and maturation closely resemble those of humans. This study compared two genome-editing approaches to disrupt GJB2 in porcine oocytes before fertilization: (1) electroporation with CRISPR/Cas9 ribonucleoprotein and (2) microinjection with cytosine base editor (BE3) and single-guide RNAs (sgRNAs). Electroporation produced high mutation rates (70-90%) across three concentrations of Cas9/sgRNA but yielded mostly heterozygous or mosaic blastocysts, with limited homozygous knockouts (< 4%). BE3 achieved precise cytosine-to-thymine conversions that introduced premature stop codons, reaching up to 47% total editing and 20% homozygous nonsense alleles. However, blastocyst formation declined at higher component concentrations. Overall, BE3 produced more predictable mutations than conventional CRISPR/Cas9, although embryo developmental competence was dose-dependent. Both methods effectively targeted GJB2 and demonstrated feasibility of pre-fertilization genome editing in porcine oocytes. These findings establish the groundwork for generating GJB2-deficient pigs as translational models of Cx26-related congenital deafness and for future evaluation of gene-therapy strategies in a large-animal system.

Monitoring contamination of hazardous drug compounding surfaces at hospital pharmacy departments. A consensus Statement. Practice guidelines of the Spanish Society of Hospital Pharmacists (SEFH)

2026-05-13T11:04:53Z

Monitoring contamination of hazardous drug compounding surfaces at hospital pharmacy departments. A consensus Statement. Practice guidelines of the Spanish Society of Hospital Pharmacists (SEFH) Valero-García, Silvia; González-Haba, Eva; Queralt-Gorgas, María; Alonso-Herreros, José-María; Cercos-Lleti, Ana-Cristina; Poveda-Andrés, José-Luis; Calleia-Hernández, Miguel-Ángel; Delgado-Sánchez, Olga OBJECTIVE: To establish a series of recommendations based on available evidence for monitoring surface contamination in the areas devoted to compounding hazardous drugs in pharmacy departments. METHOD: Based on a literature search in the Medline and Embase databases (search period: January 2009 to July 2019), as well as on a review of standards and recommendations issued by different healthcare organizations, a committee of experts from the Spanish Society of Hospital Pharmacists defined a series of safe practices for handling hazardous drugs and monitoring compounding work surfaces. Recommendation decisions were adopted by consensus among the members of the expert group, considering the recommendations reviewed, the monitoring situation in Spanish hospital departments, and the associated costs. RESULTS: Ten recommendations were formulated, structured into eight sections. They include aspects related to the drugs to be monitored; the areas to be monitored; when samples should be taken; risk determination and preparation of a sampling protocol; analytical techniques; contamination thresholds; and design of an action plan based on the sampling and decontamination results obtained. CONCLUSIONS: Surface monitoring allows hazardous drugs detection and evaluation of the effectiveness of current protocols for the safe handling of such drugs in hospital pharmacy departments. The evaluation should include an analysis of the efficacy of engineering controls, work practices and cleaning and decontamination processes.

The importance of quality control in raw materials used in pharmaceutical formulations

2026-05-13T11:04:52Z

The importance of quality control in raw materials used in pharmaceutical formulations Ramos-Martínez, Beatriz; Alonso-Herreros, José-María; Martin-de-Rosales-Cabrera, Ana-María