Wolbachia Infection through Hybridization to Enhance an Incompatible Insect Technique-Based Suppression of Aedes albopictus in Eastern Spain.

The emergence of insecticide resistance in arbovirus vectors is putting the focus on the development of new strategies for control. In this regard, the exploitation of Wolbachia endosymbionts is receiving increasing attention due to its demonstrated effectiveness in reducing the vectorial capacity of Aedes mosquitoes. Here, we describe the establishment of a naïve Wolbachia infection in a wild Aedes albopictus population of eastern Spain through a hybridization approach to obtain males capable of sterilizing wild females. The obtained lines were compared with the Wolbachia donor, Ae. albopictus ARwP, previously artificially infected with Wolbachia wPip, regarding immature and adult survival, female fecundity, egg fertility, and level of induced sterility. Our results did not show significant differences between lines in any of the biological parameters analyzed, indicating the full suitability of the hybrids to be used as a control tool against Ae. albopictus. In particular, hybrid males induced 99.9% sterility in the eggs of wild females without the need for any preliminary treatment. Being harmless to non-target organisms and the environment, the use of this bacterium for the control of Ae. albopictus deserves further exploration. This is especially relevant in areas such as eastern Spain, where this mosquito species has recently spread and may represent a serious threat due to its competence as a vector for dengue, chikungunya, and Zika viruses.

Beneficios del uso de un algoritmo de pruebas recomendadas para el diagnóstico y manejo dietético-nutricional del paciente con trastornos gastrointestinales crónicos / Benefits of using a recommended testing algorithm for the diagnosis and dietary-nutritional management of patients with chronic gastrointestinal disorders

Introducción y objetivo: los trastornos gastrointestinales crónicos como la enfermedad celiaca y la intolerancia a la lactosa o fructosa en la edad adulta son cada vez más frecuentes y se suelen acompañar de sintomatología que repercute en las actividades diarias y limita en gran medida la dieta. El espectro de síntomas que manifiestan los afectados es heterogéneo y poco específico y, además, no existe un protocolo estandarizado y consensuado para el manejo dietético, lo que dificulta un correcto diagnóstico y un adecuado tratamiento. Los trastornos relacionados con malabsorción/intolerancia alimentaria pueden originarse por causas primarias (genéticas) o secundarias (parásitos, alergias, enfermedad inflamatoria intestinal, fármacos, etc.). El empleo de análisis genéticos permite descartar o confirmar causas primarias y, cuando sea necesario, centrar la búsqueda en las secundarias. El objetivo del enfoque algorítmico que proponemos es guiar el manejo dietético-nutricional del paciente con trastornos gastrointestinales crónicos para optimizar el proceso diagnóstico y el tratamiento nutricional. Material y métodos: tras realizar una revisión bibliográfica sobre las patologías más frecuentemente asociadas a estos trastornos, se proponen un algoritmo de pruebas y los sucesivos pasos a seguir en función de los resultados obtenidos, para concretar el diagnóstico y el tratamiento. Resultados: el algoritmo propuesto pretende ser una herramienta para el personal sanitario (gastroenterólogos, endocrinólogos, nutricionistas,etc.) que atiende a este tipo de paciente. Se busca guiar el flujo de pruebas diagnósticas en función de la información aportada por el paciente y la clínica al inicio, así como recomendar el tratamiento (dietético-nutricional y/o farmacológico) más adecuado. Conclusiones: ... (AU)

Introduction and objective: chronic gastrointestinal disorders such as celiac disease and lactose or fructose intolerance in adulthood are becoming more frequent and are usually accompanied by symptoms that affect daily activities and greatly limit diet. The spectrum of symptoms manifested by those affected is heterogeneous and not very specific; in addition, there is no standardized and agreed protocol for dietary management, which makes a correct diagnosis and effective treatment difficult. Disorders related to malabsorption/food intolerance can originate from primary (genetic) or secondary causes (parasites, allergies, inflammatory bowel disease, drugs, etc.). Using genetic data makes it possible to rule out or confirm primary causes, and when necessary, focus the search on secondary ones. The objective of this algorithmic approach is to guide the dietary-nutritional management of the patient with chronic gastrointestinal disease to optimize the diagnostic process and nutritional treatment. Material and methods: after a review of the literature on the pathologies most frequently associated with these disorders, a testing algorithm is proposed and the successive steps to be followed depending on the results obtained, in order to determine the diagnosis and treatment.Results: the proposed algorithm aims to be a tool for health personnel (gastroenterologists, endocrinologists, nutritionists, etc.) who care for these patients. The aim is to guide the flow of diagnostic tests based on the information provided by the patient and the clinic at the beginning, as well as to recommend the most appropriate treatment (dietary-nutritional and/or pharmacological). Conclusions: ... (AU)

Assessment of the Health Status of Spanish Schoolchildren Based on Nutrimetry, Lifestyle and Intestinal Parasites.

Malnutrition in Spanish schoolchildren, and its relationship with lifestyles, has been studied, but Nutrimetry (a nutritional status indicator), and data on intestinal parasitism and its risk factors, have never before been taken into account. A total of 206 children aged 3-11 years, from two schools in the Valencian Community, participated. Demographic characteristics, diet, lifestyles, behavioural habits and anthropometric (weight, height) and coproparasitological data were collected. Nutrimetry was used to analyse nutritional status. Statistical analyses were performed to ascertain associations between lifestyle, selected parasite species and nutritional status. Multivariate logistic regression analysis was used to assess the strength of the association of the suspected risk factors with the presence of intestinal parasitism. The prevalence of overweight was 32.6%. A total of 43.9% had a high adherence to the Mediterranean Diet, for which mean daily intake was 2428.7 kcal. Intestinal parasitism was identified in 49.5% of the children (Giardia duodenalis: 28.6%). The source of drinking water was found to be a risk factor for intestinal parasitism. No positive association between the variables analysed and nutritional status could be confirmed. Nutrimetry is a good indicator for a complete analysis of nutritional status. It highlights the prevalence of overweight. Intestinal parasitism was identified in almost half of the participants and is a variable that should not be underestimated.

Prevalence and associated risk factors of intestinal parasites among schoolchildren in Ecuador, with emphasis on the molecular diversity of Giardia duodenalis, Blastocystis sp. and Enterocytozoon bieneusi.

BACKGROUND: Intestinal helminths, including Soil-Transmitted Helminth (STH), and Gastrointestinal Protist (GP) infections are major contributors to the global burden of disease, particularly in low-income countries such Ecuador. Their epidemiology in these settings is largely unknown. METHODOLOGY: This prospective cross-sectional study investigates the carriage of intestinal helminths, including STH, and GP in asymptomatic schoolchildren (3-11 years) in the Chimborazo and Guayas provinces, Ecuador. Single stool samples (n = 372) and epidemiological questionnaires on demographics and potential risk factors were collected from participating schoolchildren. Conventional microscopy examination was used as screening method, and molecular (PCR and Sanger sequencing) assays were used to further investigate the epidemiology of some GP. A multivariate logistic regression analysis was used to evaluate the strength of the association of suspected risk factors with the presence of helminths and GP. PRINCIPAL FINDINGS: At least one intestinal parasite species was observed by microscopy in 63.2% (235/372) of the participating schoolchildren. Enterobius vermicularis (16.7%, 62/372; 95% CI: 13.0-20.9) and Blastocystis sp. (39.2%, 146/372; 95% CI: 34.2-44.2) were the most prevalent among helminths and GP, respectively. Assemblages A (50.0%), B (37.5%) and A+B (12.5%) were detected within Giardia duodenalis and ST3 (28.6%), ST1 and ST2 (26.2% each), and ST4 (14.3%) within Blastocystis sp. Three genotypes, two known (A: 66.7%; KB-1: 16.7%) and a novel (HhEcEb1, 16.7%) were identified within Enterocytozoon bieneusi. Municipality of origin, household overcrowding, and poor sanitation and personal hygiene habits were risk factors for childhood intestinal parasites colonization. CONCLUSIONS/SIGNIFICANCE: Despite massive government drug administration programs, STH and GP infection remain a public health concern in paediatric populations living in poor-resource settings. Molecular analytical methods are required to better understand the epidemiology of these intestinal parasites. This study provides novel information on the occurrence of Blastocystis sp. and E. bieneusi genetic variants circulating in Ecuadorian human populations.

Extracellular vesicles from the trematodes Fasciola hepatica and Dicrocoelium dendriticum trigger different responses in human THP-1 macrophages.

Extracellular vesicles (EVs) released by the helminths Dicrocoelium dendriticum and Fasciola hepatica are important modulators of the host immune response, contributing to the establishment of the infection. Monocytes and, in particular, macrophages are major regulators of the inflammatory response and are likely responsible for the phagocytosis of most of the parasite EVs. In this study, we isolated EVs from F. hepatica (FhEVs) and D. dendriticum (DdEVs) by size exclusion chromatography (SEC) and characterized them by nanoparticle tracking analysis, transmission electron microscopy and LC-MS/MS, and analyzed the cohort of proteins. The treatment of monocytes/macrophages with FhEVs, DdEVs or EV-depleted fractions from SEC, demonstrated species-specific effects of the EVs. In particular, FhEVs reduce the migratory capacity of monocytes and the analysis of the cytokine profile showed that they induce a mixed M1/M2 response, exerting anti-inflammatory properties in Lipopolysaccharide-activated macrophages. In contrast, DdEVs do not affect monocyte migration and seem to have pro-inflammatory properties. These results correlate with the differences in the life cycle of both parasites, suggesting different host immune responses. Only F. hepatica migrates to the bile duct through the liver parenchyma, driving the host immune response to heal deep erosions. Furthermore, the proteomic analysis of the macrophages upon FhEV treatment identified several proteins that might be involved in FhEV-macrophage interactions.

[Wolbachia pipientis infections in populations of Aedes albopictus in the city of València (Spain): implications for mosquito control.] / Infecciones por Wolbachia pipientis en poblaciones de Aedes albopictus en la ciudad de València (España): implicaciones para el control de mosquitos.

OBJECTIVE: The presence of Aedes albopictus, of high sanitary and social impact, was first reported in Valencia (Eastern Spain) in 2015. Innovative tools for its control include the use of the endosymbiotic bacterium Wolbachia pipientis. The release of mosquito males infected with the wPip strain, has proven very promising for large-scale Incompatible Insect Technique (IIT) applications. Before this strategy can be implemented in Valencia, it is important to know whether the natural local mosquito populations are Wolbachia-infected and, if so, identifying the infecting strains/supergroups, these being the objectives of the present work. METHODS: Eggs were collected from the 19 districts of the València city between May and October 2019. A total of 50 lab-reared adult Ae. albopictus individuals were processed and analyzed for Wolbachia detection and molecular characterization. These actions took place within the framework of a collaboration established with the Department of Health and Consumer Affairs of the city council of Valencia. Fisher's exact test was used to detect the statistical significance of the differences between groups. RESULTS: Our study revealed that 94% of the analyzed samples were naturally infected with Wolbachia. Both wAlbA and wAlbB supergroups were identified, with most samples (72% of the infected ones) carrying co-infections. CONCLUSIONS: These data provide the first characterization of the Wolbachia presence in natural populations of Ae. albopictus in the Mediterranean area of Spain. This information is relevant to evaluate the potential use of Wolbachia strains in order to achieve the suppression of the Asian tiger mosquito populations through massive release of artificially-infected males.

OBJETIVO: La presencia de Aedes albopictus, de alto impacto sanitario y social, se informó por primera vez en Valencia en 2015. Las herramientas innovadoras para su control incluyen el uso de la bacteria endosimbiótica Wolbachia pipientis. La liberación de mosquitos machos infectados con la cepa wPip ha demostrado ser muy prometedora para aplicar la Técnica de Insectos Incompatibles (IIT) a gran escala. Antes de que esta estrategia pueda implementarse, es importante saber si las poblaciones locales de mosquitos silvestres están infectadas por Wolbachia y, de ser así, identificar las cepas/supergrupos infectantes, siendo estos los objetivos del presente trabajo. METODOS: Se recolectaron huevos de los diecinueve distritos de València entre mayo y octubre de 2019, y se mantuvieron en el laboratorio hasta llegar a adultos. Un total de cincuenta individuos adultos de Ae albopictus fueron procesados y analizados para detectar la presencia de Wolbachia y su caracterización molecular. Estas acciones se enmarcaron en la colaboración establecida con la Concejalía de Salud y Consumo del Ayuntamiento de València. La prueba exacta de Fisher fue utilizada para detectar la significación estadística de las diferencias entre grupos. RESULTADOS: El 94% de las muestras analizadas estaban infectadas de forma natural con Wolbachia. Se identificaron los supergrupos wAlbA y wAlbB, y la mayoría de las muestras (72% de las infectadas) presentaban coinfecciones. CONCLUSIONES: Los datos proporcionan la primera caracterización de la presencia de Wolbachia en poblaciones naturales de Ae. albopictus en el área mediterránea de España. Esta información es relevante para evaluar el potencial uso de cepas de Wolbachia de cara a la supresión de poblaciones de mosquito tigre asiático mediante la liberación masiva de machos infectados artificialmente.

Infecciones por Wolbachia pipientis en poblaciones de Aedes albopictus en la ciudad de València (España): implicaciones para el control de mosquitos / Wolbachia pipientis infections in populations of Aedes albopictus in the city of València (Spain): implications for mosquito control

Fundamentos: La presencia deAedes albopictus, de alto impacto sanitario y social, se informó por primera vez en Valenciaen 2015. Las herramientas innovadoras para su control incluyen el uso de la bacteria endosimbióticaWolbachia pipientis. La liberación de mosquitos machos infectados con la cepawPip ha demostrado ser muy prometedora para aplicar la Técnica de InsectosIncompatibles (IIT) a gran escala. Antes de que esta estrategia pueda implementarse, es importante saber si las poblaciones localesde mosquitos silvestres están infectadas porWolbachia y, de ser así, identificar las cepas/supergrupos infectantes, siendo estos losobjetivos del presente trabajo. Métodos: Se recolectaron huevos de los diecinueve distritos de València entre mayo y octubre de 2019, y se mantuvieron en ellaboratorio hasta llegar a adultos. Un total de cincuenta individuos adultos deAe. albopictus fueron procesados y analizados paradetectar la presencia deWolbachia y su caracterización molecular. Estas acciones se enmarcaron en la colaboración establecida conla Concejalía de Salud y Consumo del Ayuntamiento de València. La prueba exacta de Fisher fue utilizada para detectar la significaciónestadística de las diferencias entre grupos. Resultados: El 94% de las muestras analizadas estaban infectadas de forma natural conWolbachia. Se identificaron los supergruposwAlbA ywAlbB, y la mayoría de las muestras (72% de las infectadas) presentaban coinfecciones. Conclusiones: Los datos proporcionan la primera caracterización de la presencia deWolbachia en poblaciones naturales deAe. albopictus en el área mediterránea de España. Esta información es relevante para evaluar el potencial uso de cepas deWolbachiade cara a la supresión de poblaciones de mosquito tigre asiático mediante la liberación masiva de machos infectados artificialmente.(AU)

Background: The presence ofAedes albopictus, of high sanitary and social impact, was first reported in Valencia (Eastern Spain)in 2015. Innovative tools for its control include the use of the endosymbiotic bacteriumWolbachia pipientis. The release of mosquitomales infected with thewPip strain, has proven very promising for large-scale Incompatible Insect Technique (IIT) applications. Beforethis strategy can be implemented in Valencia, it is important to know whether the natural local mosquito populations areWolbachia-infected and, if so, identifying the infecting strains/supergroups, these being the objectives of the present work. Methods: Eggs were collected from the 19 districts of the València city between May and October 2019. A total of 50 lab-reared adultAe. albopictus individuals were processed and analyzed forWolbachia detection and molecular characterization. These actions tookplace within the framework of a collaboration established with the Department of Health and Consumer Affairs of the city council ofValencia. Fisher’s exact test was used to detect the statistical significance of the differences between groups. Results: Our study revealed that 94% of the analyzed samples were naturally infected withWolbachia. BothwAlbA andwAlbBsupergroups were identified, with most samples (72% of the infected ones) carrying co-infections. Conclusions: These data provide the first characterization of theWolbachia presence in natural populations ofAe. albopictusin the Mediterranean area of Spain. This information is relevant to evaluate the potential use ofWolbachia strains in order to achievethe suppression of the Asian tiger mosquito populations through massive release of artificially-infected males.(AU)

Field study of parasitic contamination of fruits, vegetables and leafy greens in the Ecuadorian Andes.

Background: Raw vegetables have been considered vehicles of enteroparasites. South American countries are among the most important exporters of fresh vegetables, including Ecuador, which has a tropical climate and soils rich in organic matter that allow it to harvest throughout the year for sale to different countries. The aim of the study was to assess the occurrence of the parasitic contamination of fruits, vegetables and leafy greens grown in an agricultural area of the Ecuadorian Andes. Methods: A cross-sectional field study was conducted with snowball sampling on 1,416 samples (516 fruits, 488 vegetables, and 412 leafy greens). Each sample were washed with water, and the resulting solution after removing the vegetables, was subjected to 24-hour sedimentation. The concentrated sediment underwent microscopic analysis. Results: The overall positivity for parasitic contamination was 63.4%, with leafy greens having the highest contamination rate (76.9%) (P<0.0001), surpassing vegetables (67.8%) and fruits (48.4%). Cabbage (100%), onions (84%), and strawberries (60.2%) emerged as the most contaminated within their respective groups. Protozoa were more prevalent (49.6%) than helminths (15.5%) (P<0.0001). Blastocystis sp. (33.5%) ranked highest, followed by Eimeria spp. (26.3%), Entamoeba spp. (10.3%), Giardia spp. (8.3%), Balantidium spp. (6.9%), Cryptosporidium spp. (6.6%), Cyclospora spp. (4.4%), Cystoisospora spp. (0.5%), Strongylida (15.5%), and Ascaris spp. (0.4%). Conclusions: The study reveals that vegetables and fruits for human consumption from this area of the Ecuadorian Andes are highly contaminated with various parasites, constituting a possible source of infection for humans and animals in this area, or in non-endemic areas where these products are marketed. The finding emphasizes the need for strict hygienic measures in agricultural crops, which will be properly achieved through the treatment of soil, manure and water used for cultivation.

Parasite Fauna and Coinfections in Urban Rats Naturally Infected by the Zoonotic Parasite Angiostrongylus cantonensis.

When the zoonotic parasite of rodents that can cause human neuroangiostrongyliasis, i.e., Angiostrongylus cantonensis, is found in its natural definitive hosts, it is usually reported in isolation, as if the rat lungworm were the only component of its parasite community. In this study, we report the coinfections found in rats naturally infected by A. cantonensis in urban populations of Rattus norvegicus and Rattus rattus in Valencia, Spain. In addition to the rat lungworms, which were found in 14 of the 125 rats studied (a prevalence of 11.20%), 18 other parasite species (intestinal and tissular protists, microsporidia and helminths) were found, some of them with high burdens. Fourteen of these nineteen species found are potential zoonotic parasites, namely Blastocystis, Giardia duodenalis, Cryptosporidium spp., Enterocytozoon bieneusi, Encephalitozoon hellem, Toxoplasma gondii, Brachylaima spp., Hydatigera taeniaeformis s.l. larvae, Hymenolepis nana, Hymenolepis diminuta, Angiostrongylus cantonensis, Calodium hepaticum, Gongylonema neoplasticum and Moniliformis moniliformis. The total predominance of coinfected rats as well as their high parasite loads seem to indicate a trend towards parasite tolerance.

Do Intestinal Unicellular Parasites Have a Role in the Inflammatory and Redox Status among the Severely Obese?

The diagnosis of obesity comprises subjects with totally different phenotypes and metabolic profiles. Systemic inflammation and oxidative stress derived from the white adipose tissue are suggested as the link between this disease and the development of insulin resistance and metabolic comorbidities. The presence of unicellular eukaryotic parasites colonizing the human gut ecosystem is a common circumstance, and yet their influence on the inflammatory and redox status of the obese host has not been assessed. Herein, a set of inflammatory and redox biomarkers were assessed together with a parasitological analysis of 97 severely obese subjects. Information was also collected on insulin resistance and on the antioxidant composition of the diet. The global prevalence of intestinal unicellular parasites was 49.5%, with Blastocystis sp. the most prevalent protozoan found (42.3%). Colonized subjects displayed a higher total antioxidant capacity and a trend towards higher extracellular superoxide dismutase activity, regardless of their insulin resistance status, along with lower reduced glutathione/oxidized glutathione (GSH/GSSG) ratios in plasma in the insulin-resistant subgroup. No changes in malondialdehyde levels, or in inflammatory cytokines in plasma, were found in regard to the colonization status. In conclusion, enteric eukaryotic unicellular parasites may play an important role in modulating the antioxidant defenses of an obese host, thus could have beneficial effects with respect to the development of systemic metabolic disorders.

Evaluation of School Children Nutritional Status in Ecuador Using Nutrimetry: A Proposal of an Education Protocol to Address the Determinants of Malnutrition.

The education sector is a cornerstone in the battle against malnutrition in children. However, there are still no consolidated protocols that outline strategies for how nutrition programs in low- and middle-income countries can be delivered through the education sector. Establishing the correct community diagnosis is essential prior to the elaboration of an intervention plan for a school population that takes into account more than just traditional variables related to the nutritional status. A total of 574 boys and girls aged 3-11 years from three educational institutions in different municipalities in Ecuador participated in the study. Sociodemographic, anthropometric (weight and height) and coproparasitological data were obtained. Nutrimetry, which is a combination of two classical anthropometrics indicators, was used for the analysis of the nutritional status, and the indicators' frequencies varied among the schools. In order to improve the nutritional status of children, we proposed a framework mainly focusing on establishing alliances with the education sector and taking into account gender equality; respect for the environment; and the customs, beliefs and traditions of each population. The results obtained from the analyses of other variables demonstrated the importance of an adequate diagnosis prior to any type of intervention at the nutritional level, since characteristics could vary by local area and have an impact on the successfulness of the intervention.

Interplay between Intestinal Bacterial Communities and Unicellular Parasites in a Morbidly Obese Population: A Neglected Trinomial.

Obesity is an epidemic causing a metabolic health crisis. Herein, the interactions between the gut prokaryotic and eukaryotic communities, metabolic comorbidities and diet were studied. Stool samples from 56 subjects, 47 with type III obesity and 9 with type II obesity and cardiovascular risk or metabolic disease, were assessed for the richness, diversity and ecology of the bacterial gut community through metagenomics, together with the study of the presence of common unicellular eukaryote parasites (Blastocystis sp., Dientamoeba fragilis and Giardia intestinalis) by qPCR. Clinical information regarding metabolic comorbidities and non-alcoholic hepatic fatty liver disease was gathered. To assess the quality of the patients' diet, each participant filled in three dietary questionnaires. The most prevalent parasite Blastocystis sp. (46.4%), together with D. fragilis (8.9%), was found to be associated with higher mean diversity indexes regarding non-colonized subjects; the opposite of that which was observed in those with G. intestinalis (16.1%). In terms of phyla relative abundance, with Blastocystis sp. and D. fragilis, very slight differences were observed; on the contrary, G. intestinalis was related to an increase in Bacteroidetes and Proteobacteria, and a decrease in Firmicutes and Actinobacteria, presenting the lowest Firmicutes/Bacteroidetes ratio. At genus level, Blastocystis sp. and/or D. fragilis was accompanied with an increase in Lactobacillus spp., and a decrease in Akkermansia spp., Bifidobacterium spp. and Escherichia spp., while G. intestinalis was associated with an increase in Bacteroides spp., and a decrease in Faecalibacterium spp., Prevotella spp. and Lactobacillus spp., and the highest Bacteroides spp./Prevotella spp. ratio. Participants with non-alcoholic hepatic fatty liver presented a higher Firmicutes/Bacteroidetes ratio, and those with type 2 diabetes displayed a significantly lower Faecalibacterium spp./Escherichia spp. ratio, due to an overrepresentation of the genus Escherichia spp. The presence of parasites was associated with variations in the richness, diversity and distribution of taxa in bacterial communities, confirming a gain in diversity associated with Blastocystis sp. and providing different functioning of the microbiota with a potential positive effect on comorbidities such as type 2 diabetes, insulin resistance and metabolic syndrome. Future basic and clinical studies should assess the beneficial or pathogenic effect of these eukaryotes on obese subjects and focus on deciphering whether they may imply a healthier metabolic profile.

Evaluation of the Immunomodulatory Effect of the Recombinant 14-3-3 and Major Antigen Proteins of Strongyloides stercoralis against an Infection by S. venezuelensis.

Strongyloidiasis, caused by Strongyloides stercoralis, is a neglected parasitic disease that represents a serious public health problem. In immunocompromised patients, this parasitosis can result in hyperinfection or disseminated disease with high levels of mortality. In previous studies, the mRNAs encoding for the 14-3-3 and major antigen proteins were found to be expressed at high levels in S. stercoralis L3 larvae, suggesting potential key roles in parasite-host interactions. We have produced them as recombinant proteins (rSs14-3-3 and rSsMA) in a bacterial protein expression system. The serum levels of anti-rSs14-3-3 and anti-rSsMA IgGs are increased upon infection with S. venezuelensis, validating the use of the mouse model since the native 14-3-3 and MA proteins induce an immune response. Each recombinant protein was formulated in the adjuvant adaptation (ADAD) vaccination system and injected twice, subcutaneously, in CD1 mice that were experimentally infected with 3000 S. venezuelensis L3 to evaluate their protective and immunomodulatory activity. Our results, including the number of parthenogenetic females, number of eggs in stool samples and the analysis of the splenic and intestinal indexes, show that the vaccines did not protect against infection. The immunization with rSs14-3-3 induced changes in the cytokine profile in mice, producing higher expression of IL-10, TGF-ß, IL-13 and TNF-α in the spleen, suggesting a Th2/Treg-type response with an increase in TNF-α levels, confirming its role as an immunomodulator.

Survey of the occurrence of Giardia duodenalis cysts and Cryptosporidium spp. oocysts in green leafy vegetables marketed in the city of Valencia (Spain).

The role of vegetables usually consumed without prior culinary treatment is known to contribute to the prevalence of foodborne diseases. Cysts and oocysts can contaminate food, which can then be the source of infection in humans. The aim of the study was to assess the occurrence of Giardia duodenalis and Cryptosporidium spp. (oo)cysts in green leafy vegetables marketed in the city of Valencia (Spain) combining parasitological methods, two real-time qPCRs and light microscopy. An experimental field study was conducted on 129 vegetable samples, 64 from conventional farms and 65 from ecological (organic) farms. The samples were washed with water, and the resulting solution after removing the vegetables, was subjected to 24-hour sedimentation. The concentrated sediment was used for the search for protozoa. A positive result by both real-time PCRs, or a positive result by one qPCR and confirmation by microscopy was established as a positivity criterion. Giardia duodenalis was detected in 23.0 % of the samples, and Cryptosporidium spp. in 7.8 %. G. duodenalis (41.5 %) and Cryptosporidium spp. (20.0 %) were more frequent in ecological crops. The high level of contamination detected in organic vegetables may be due to the type of fertilizers and the quality of the water used for their irrigation and reinforces the need to take extreme hygiene measures in vegetables that are consumed raw.

Nematode Parasites of the European Pilchard, Sardina pilchardus (Walbaum, 1792): A Genuine Human Hazard?

The European pilchard is one of the most frequently consumed fish species in Mediterranean countries, especially in Italy and Spain, and has been reported as the cause of at least eight human anisakidosis cases in Spain. With the aim to shed light on the potential human parasitosis risk posed by nematode larvae belonging to families Anisakidae or Raphidascarididae, a total of 350 sardines captured in the Atlantic Ocean (175 specimens) and the Mediterranean Sea (175 specimens), acquired in various Spanish nationwide supermarket chains, were helminthologically analyzed. The statistical analysis of some helminth parameters revealed a higher presence of nematodes belonging to the genus Hysterothylacium (prevalence 24.29%; mean abundance of 2.36), usually considered non-parasitic for humans (only three cases reported worldwide), when compared to nematodes of the genus Anisakis (5.71%; 0.16). The human anisakidosis risk after the consumption of raw or undercooked sardines and the role of Hysterothylacium, the most frequent nematode, is discussed, providing information to consumers. To avoid human infection by anisakid larval nematodes, the established preventive measures are confirmed and new ones are proposed, such as the consumption of sardines preferably caught in the Mediterranean and of small-sized specimens available, and the immediate evisceration after fishing.

Influence of Environmental Pollution and Living Conditions on Parasite Transmission among Indigenous Ecuadorians.

The purpose of this study was to evaluate the influence of environmental pollution and the living conditions of indigenous Ecuadorians on the transmission of enteroparasites in an Andean agricultural area located at high altitude. Environmental pollution was recorded after observation in each community. The parasites were identified by microscopic sediment analysis using physiological saline solution from macerated arthropods, washed vegetables, and human stools, utilizing four coproparasitological techniques (direct examination, Kato-Katz, ether concentration, and Ziehl-Neelsen). The results show that the inadequate disposal of human and animal excreta that contaminate soil and water, incorrect food hygiene, inadequate sanitary infrastructure in houses, a lack of animal veterinary care, and rodent proliferation are important reservoirs of zoonotic parasites. The use of excrement as fertilizer increases the number of flies, which act as mechanical vectors, and vegetables grown in areas with disperse infective parasitic forms act as vehicles that are marketed at the local, regional, and international levels. These analyses verify contamination levels of 52.7% in mechanical vectors, 70.6% in vegetables, and 98.2% in human stools. The agricultural communities analyzed maintained poor hygienic-sanitary and environmental conditions, which had a significant influence on the transmission of enteroparasites that affect human health.

Anisakid Nematodes and Potential Risk of Human Anisakiasis through the Consumption of Hake, Merluccius spp., Sold Fresh in Spanish Supermarkets.

Nematode parasite species belonging to the Anisakis simplex complex are the most important cause of human anisakiasis through the consumption of (mainly) undercooked, previously not frozen, or conveniently treated fish. In Spain, the consumption of hake has been recognized as an important source of this parasitosis. With the aim of shedding light on the risk factors that can influence the potential risk of human anisakiasis in Spain through the consumption of fresh hake sold by nationwide supermarket chains, a total of 536 small hake specimens belonging to the species Merluccius bilinearis caught off the Northeast American coasts and Merluccius merluccius caught in the Northeast Atlantic and Mediterranean waters was analysed. Anisakids morphologically identified as Anisakis type I were found as the most prevalent and the most abundant species and were considered the main potential cause of human anisakiasis. Intrinsic and extrinsic factors concerning the hake, such as its origin and season of capture, its size, as well as the days passed between its capture and consumption, should be taken into account to avoid this human parasitosis. It is essential that consumers have access to fish label information which should include, as regulated by the European Commission, traceability data.

Proteomic Analysis of Extracellular Vesicles From Fasciola hepatica Hatching Eggs and Juveniles in Culture.

The identification of extracellular vesicles (EVs) in Fasciola hepatica has provided a new way to understand parasite-host communication. Most of the studies on EVs have focused on the adult stage of F. hepatica, but recently, the presence of EVs from different developmental stages has been reported. To better understand the potential role of EVs in the biology of the parasite and in the infection process, the protein cargo of EVs from embryonated eggs and newly-excysted juvenile (NEJs) flukes cultured up to 28 days, has been analyzed. EVs were isolated by size exclusion chromatography and evaluated by nanoparticle tracking analysis and transmission electron microscopy. LC-MS/MS proteomic analysis of EVs revealed the presence of 23 different proteins from embryonated egg-derived EVs and 29 different proteins from NEJ-derived EVs. Most of the identified proteins had been previously described in EVs from F. hepatica adults, including cytoskeletal proteins, glycolytic enzymes, stress-related proteins and tetraspanins. Nevertheless, EVs from hatching eggs and NEJs exhibited qualitative differences in composition, when compared to EVs form adults, including the absence of cathepsin cysteine peptidases. The differential content of the EVs released by the different developmental stages of the parasite reflect the intense activity of NEJs at this early stage, with several proteins involved in membrane traffic and cell physiology. This new set of identified proteins could help to understand key metabolic, biochemical and molecular mechanisms mediated by EVs that take place upon egg hatching and after parasite excystment.

Presence of Blastocystis in gut microbiota is associated with cognitive traits and decreased executive function.

Growing evidence implicates the gut microbiome in cognition. Blastocystis is a common gut single-cell eukaryote parasite frequently detected in humans but its potential involvement in human pathophysiology has been poorly characterized. Here we describe how the presence of Blastocystis in the gut microbiome was associated with deficits in executive function and altered gut bacterial composition in a discovery (n = 114) and replication cohorts (n = 942). We also found that Blastocystis was linked to bacterial functions related to aromatic amino acids metabolism and folate-mediated pyrimidine and one-carbon metabolism. Blastocystis-associated shifts in bacterial functionality translated into the circulating metabolome. Finally, we evaluated the effects of microbiota transplantation. Donor's Blastocystis subtypes led to altered recipient's mice cognitive function and prefrontal cortex gene expression. In summary, Blastocystis warrant further consideration as a novel actor in the gut microbiome-brain axis.

Ocurrence of enteroparasites with zoonotic potential in animals of the rural area of San Andres, Chimborazo, Ecuador.

OBJECTIVE: The aim of this research was the identification of the enteroparasites harbored by the animals of the San Andrés community, to evaluate their role as susceptible hosts and sources of infection for other animals, humans (zoonoses), as well as parasite forms spreaders to the environment in this rural area, located in the province of Chimborazo, Ecuadorian Andean region. MATERIAL AND METHODS: The study was carried out combining 3 coproparasitological techniques: direct examination, Ritchie and Ziehl-Neelsen in 300 animal stool samples RESULTS: Blastocystis sp., Entamoeba spp., Giardia spp., Balantidium spp., Cryptosporidium spp., Ascaris spp., Toxocara spp., Ancylostoma spp., Strongylida, Hymenolepis nana and Echinococcus spp., were detected. Infection by protozoa (87.3%) was higher than helminths (31.0%). All cattle, sheep and guinea pigs were found parasitized, and the presence of Blastocystis sp., Entamoeba spp. and Cryptosporidium spp. by all groups of animals stands out. It is also remarkable the presence of Giardia spp. in swine (19.2%), big herbivores-livestock (11.5%), leporids (8.3%) and carnivores (5.9%); Balantidium spp. in swine (19.2%), big herbivores-livestock (5.8%) and carnivores (1.2%); Hymenolepis nana in guinea pigs (2.1%); and Toxocara spp. (15.7%), Echinococcus spp. (9.6%) and Ancylostoma spp. (6.0%) in dogs. CONCLUSION: Animals from San Andrés have a wide spectrum of intestinal parasitic forms in their feces, being a source of infection to other animals and humans, and a source of contamination of the environment, posing a risk factor and reinforcing the idea of the need for more effective treatments and hygienic measures to improve livestock production and cutting its transmission.