Detection and genotyping of zoonotic microsporidia in the endangered Iberian lynx (Lynx pardinus).

Microsporidia is a diverse group of obligate, intracellular, and spore-forming parasites that infect a wide range of animals. Among them, Enterocytozoon bieneusi and Encephalitozoon spp. are the most frequently reported species in humans. Limited information is available about the presence and molecular diversity of microsporidian species in the endangered Iberian lynx (Lynx pardinus). Presence of Enterocytozoon bieneusi and Encephalitozoon spp. was investigated by molecular methods in wild and captive Iberian lynxes from Spain. Overall, E. bieneusi was detected in 3.2% (8/251) of the animals examined. None of the samples tested were positive for Encephalitozoon spp. Four known (D, EbfelA, PigEBITS7, and Type IV) and a novel (named as LynxSpEb1) E. bieneusi genotypes were identified. All the genotypes found belonged to the zoonotic Group 1 of E. bieneusi. This study provides the first genotyping data of E. bieneusi in Iberian lynx in Spain. Our result indicate that the Iberian lynx does not seem to play a relevant role in the epidemiology of Encephalitozoon spp., and that this endangered felid is likely acting as spillover host rather than a true reservoir of E. bieneusi. Additional studies should be conducted to assess the impact of this parasite in the health status of the endangered Iberian lynx.

Antibacterial Effect of Spanish Honeys of Different Botanical Origins against Staphylococcus epidermidis.

Honey is traditionally used for its medicinal properties attributed to its antibacterial and antioxidant effects. It is considered a natural alternative to conventional antibiotics. This effect has been attributed to their physico-chemical properties, as various chemical parameters can synergistically influence this effect. The aim of this study is to assess Spanish honeys of diverse botanical origins for their antibacterial efficacy against Staphylococcus epidermidis, correlating their physico-chemical attributes, (poly)phenol content, and antioxidant activity. The methods included colour determination via two methodologies, acidity, pH, moisture content, and sugar concentration. (Poly)phenol content was quantified using the Folin-Ciocalteau method, while antioxidant activity was evaluated via the FRAP method. Subsequently, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against S. epidermidis were investigated with different concentrations of honeys. The results revealed a direct relationship between honey darkness, (poly)phenol concentration, antioxidant activity, and antibacterial efficacy. Darker honeys exhibited higher (poly)phenol levels, greater antioxidant activity, and consequently, lower MIC and MBC values, showing enhanced antibacterial properties. These findings underscore the potential of honey as a therapeutic agent against S. epidermidis, particularly in wound healing applications to avoid infection. Further research into honey's multifaceted properties is warranted to unveil novel therapeutic avenues in healthcare.

Chemical Composition, Functional and Antioxidant Properties of Dietary Fibre Extracted from Lemon Peel after Enzymatic Treatment.

Lemon peel represents an interesting by-product owing to its content of dietary fibre (DF) and (poly)phenols, which is of great importance for its valorisation. Hence, the objective of this study was to characterise the DF, total phenolic content (TPC), and antioxidant capacity of two lemon-peel-derived ingredients using two different methods (drying with warm air and enzymatic hydrolysis with pectinesterase). The analysis included a DF assessment, followed by neutral sugars characterisation through GC-FID and uronic acids determination via colorimetry. Subsequently, TPC and antioxidant capacity using the FRAP method were quantified through spectrophotometry. The swelling capacity (SWC), water retention capacity (WRC), and fat absorption capacity (FAC) were also determined as functional properties. It was observed that pectinesterase treatment led to a reduction in soluble DF and an increase in insoluble DF. This treatment also affected the pectin structure, thereby diminishing its ability to absorb water and fat within its matrix. The TPC was also reduced, resulting in a decrease in antioxidant capacity. Conversely, employing warm air exhibited a noteworthy increase in antioxidant capacity. This underscores its crucial contribution to the valorisation of lemon peel, not only by diminishing the environmental impact but also by enabling the acquisition of fibre ingredients with a noteworthy antioxidant capacity.

Microbial Matryoshka: Addressing the Relationship between Pathogenic Flagellated Protozoans and Their RNA Viral Endosymbionts (Family Totiviridae).

Three genera of viruses of the family Totiviridae establish endosymbiotic associations with flagellated protozoa responsible for parasitic diseases of great impact in the context of One Health. Giardiavirus, Trichomonasvirus, and Leishmaniavirus infect the protozoa Giardia sp., Trichomonas vaginalis, and Leishmania sp., respectively. In the present work, we review the characteristics of the endosymbiotic relationships established, the advantages, and the consequences caused in mammalian hosts. Among the common characteristics of these double-stranded RNA viruses are that they do not integrate into the host genome, do not follow a lytic cycle, and do not cause cytopathic effects. However, in cases of endosymbiosis between Leishmaniavirus and Leishmania species from the Americas, and between Trichomonasvirus and Trichomonas vaginalis, it seems that it can alter their virulence (degree of pathogenicity). In a mammalian host, due to TLR3 activation of immune cells upon the recognition of viral RNA, uncontrolled inflammatory signaling responses are triggered, increasing pathological damage and the risk of failure of conventional standard treatment. Endosymbiosis with Giardiavirus can cause the loss of intestinal adherence of the protozoan, resulting in a benign disease. The current knowledge about viruses infecting flagellated protozoans is still fragmentary, and more research is required to unravel the intricacies of this three-way relationship. We need to develop early and effective diagnostic methods for further development in the field of translational medicine. Taking advantage of promising biotechnological advances, the aim is to develop ad hoc therapeutic strategies that focus not only on the disease-causing protozoan but also on the virus.

Detection and Molecular Diversity of Cryptosporidium spp. and Giardia duodenalis in the Endangered Iberian Lynx (Lynx pardinus), Spain.

Cryptosporidium spp. and Giardia duodenalis are the main non-viral causes of diarrhoea in humans and domestic animals globally. Comparatively, much less information is currently available in free-ranging carnivore species in general and in the endangered Iberian lynx (Lynx pardinus) in particular. Cryptosporidium spp. and G. duodenalis were investigated with molecular (PCR and Sanger sequencing) methods in individual faecal DNA samples of free-ranging and captive Iberian lynxes from the main population nuclei in Spain. Overall, Cryptosporidium spp. and G. duodenalis were detected in 2.4% (6/251) and 27.9% (70/251) of the animals examined, respectively. Positive animals to at least one of them were detected in each of the analysed population nuclei. The analysis of partial ssu rRNA gene sequences revealed the presence of rodent-adapted C. alticolis (n = 1) and C. occultus (n = 1), leporid-adapted C. cuniculus (n = 2), and zoonotic C. parvum (n = 2) within Cryptosporidium, and zoonotic assemblages A (n = 5) and B (n = 3) within G. duodenalis. Subgenotyping analyses allowed for the identification of genotype VaA19 in C. cuniculus (gp60 locus) and sub-assemblages AI and BIII/BIV in G. duodenalis (gdh, bg, and tpi loci). This study represents the first molecular description of Cryptosporidium spp. and G. duodenalis in the Iberian lynx in Spain. The presence of rodent/leporid-adapted Cryptosporidium species in the surveyed animals suggests spurious infections associated to the Iberian lynx's diet. The Iberian lynx seems a suitable host for zoonotic genetic variants of Cryptosporidium (C. parvum) and G. duodenalis (assemblages A and B), although the potential risk of human transmission is regarded as limited due to light parasite burdens and suspected low excretion of infective (oo)cysts to the environment by infected animals. More research should be conducted to ascertain the true impact of these protozoan parasites in the health status of the endangered Iberian lynx.

Asociación de hemoglobinopatía S y alfatalasemia. Análisis de 45 casos / Association of hemoglobinopathy and alpha thalassemia. Study of 45 patients

FUNDAMENTO Y OBJETIVO: La hemoglobina (Hb) S y la alfatalasemia son, junto con la betatalasemiay las Hb C, E y D, las denominadas hemoglobinopatías comunes, las cuales presentan una alta prevalencia en las zonas históricamente endémicas de paludismo. En este estudio analizamos la frecuencia de la asociación de estas 2 hemoglobinopatías, así como su expresión fenotípica. PACIENTES Y MÉTODO: Entre enero de 1995 y marzo de 2003 hemos realizado un estudio molecular para descartar la presencia de una alfatalasemia asociada de un total de 83 casos diagnosticados de hemoglobinopatías S por criterios electroforéticos y cromatográficos. El estudio molecular se realizó por Southern blot con las endonucleasas de restricción Bam HI y Bgl II, y lassondas α (1,5 Kb Pst I) y ζ (1,8 Kb Sac Y).RESULTADOS: De los 83 casos estudiados, 45 (54,2%) presentaban alfatalasemia (37 -α3,7/αα; 1-α4,2/αα; 6 -α3,7/-α3,7; 1 -α4,2/-α4,2); 36 no tenían alfatalasemia y en 2 había una triplicación degenes α. Los pacientes con HbS heterocigota asociada a alfatalasemia presentaron el porcentajede HbS (p 50%en nuestra experiencia). El cribado de esta asociación es de gran interés, ya que puede modularla expresión clínica de los casos de HbS homocigota. Dicha asociación se debe sospecharen los casos de HbS heterocigota con valores más bajos de HbS de lo esperado

BACKGROUND AND OBJECTIVE: Hemoglobin S (HbS) α-thalassemia together with β-thalassemia and hemoglobins C, E and D are named common hemoglobinopathies. In this study, we analyzed the frequency of the association between α-thalassemia and HbS and its phenotypic expression. PATIENTS AND METHOD: Since January 1995 to March 2003 we studied 83 cases of HbS, which were diagnosed by electrophoretic and chromatographic criteria. The molecular study was carried out by Southern blot with the restriction enzymes Bam HI and Bgl II and a (1.5 Kb Pst)and z (1.8 Kb Sac Y) probes. RESULTS: 45 cases (54.2%) had a-thalassemia (36 -α3,7/aa; 1-α4,2/αα; 6 -α3,7/-α3,7; 1-α4,2/-α4,2); 36 had not α-thalassemia and in two cases there were a triplication of a genes. The patients with HbS heterozygote associated with α-thalassemia showed a percentage of HbS(p 50%). Screening for this association is of great interest because the clinical expression in the cases of HbS homozygote will be modulated. This association must be suspected in cases of homozygous HbSin which levels of HbS are lower than expected