Study of the helminth fauna in eagle owl (Bubo bubo) in the south of Spain.

In this study we show the results of the eagle owls' (Bubo bubo) helminthfauna found in Andalusia. A total number of 50 specimens have been analysed in a period of 10 years (from 2011 to 2020). Prevalence ( P % ), mean intensity (IM) and mean abundance (AM) of parasitation have been obtained. The percentage of parasitation in the total sample was 80% (40 out of 50 eagle owls): 78% nematodes, 8% trematodes, 6% cestodes and 4% acantocephalans. 7 species of helminths were identified: 6 nematodes, and 1 trematode. In the case of cestodes and acantocephalans it was not possible to determine species and only the genus was identified. The intestinal nematode Capillaria tenuissima ( P %  = 58% (44-71.2); IM = 11,52 (5.83-28.9)) was the core species whereas Synhimantus laticeps (P% = 16 (7.5-28.8); IM = 4 (1.75-7.25)) and Hartertia hispanica (P% = 16 (7.5-28.8); IM = 1,5 (1-2)) were the secondary species. The remainder species were considered satellite species, with low prevalence and average abundance. Likewise, descriptive parameters of the helminth community were determined: species richness, 1.56 (1.29-1.94), total abundance, 12 (7.24-26.40), Brillouin's diversity index, 0.18 (0.10-0.29) and Berger-Parker dominance index, 0.88 (0.81-0.93). The data from this study show a non-diverse helminthic community, without species dominance with C. tenuissima as the central species, followed by S. laticeps and H. hispanica as secondary species. Worth mentioning is the presence of H. hispanica, which is considered an endemic species in Spain and specifically in Andalusia. To the authors' knowledge, this is the largest population sample taken in parasitological studies about helminths of this raptor in Europe and the first one carried out in the south of Spain (Andalusia).

COVID 19 and Psychological Problems in Mexican Mothers

COVID-19 seriously affected people’s mental health. Possibly symptoms of anxiety, depression and insomnia occur with most frequency in various sectors of the population, especially in mothers. The objective of this research was to evaluate levels of depression, anxiety and insomnia in Mexican mothers who had COVID-19, who care for or cared for patients with COVID-19, or in neither of these two conditions. A sample of 540 Mexican mothers was gathered, a questionnaire was applied to obtain data on sociodemographic variables, as well as instruments to measure depression, anxiety and insomnia. The 70% of the participants had insomnia, 77% had depressive symptoms, while 80.2% showed anxiety. Working mothers presented more severe symptoms of insomnia than housewives. Caring for a COVID-19 patient was significantly associated with the psychological variables evaluated. Mexican mothers who care or cared for COVID-19 patients have high rates of depression, anxiety, and insomnia (AU)

Trabajadores de centros deportivos fitness percepción de liderazgo, engagement y estrés / Sports centre workers fitness effects of leadership, engagement and stress

El objetivo del estudio fue determinar la influencia del liderazgo, engagement y estrés percibido en trabajadores de centros deportivos fitness. Realizando un estudio cuantitativo, descriptivo de corte transversal. La muestra compuesta por 106 trabajadores, con una edad media de 35 años (35.2 ± 8.5). Los instrumentos utilizados fueron, Multifactor Leadership Questionnaire (MLQ-6S) para medir el tipo de liderazgo; la Utrecht Work Engagement Scale (UWES) para saber el nivel de engagement y Perceived Stress Scale (PSS), para conocer la percepción del estrés. Se comparan y correlacionan variables, t-Student, ANOVA y correlación lineal de Pearson (r) y regresión lineal múltiple. Los principales resultados resaltan que el nivel de estrés percibido es mayor en trabajadores fijos (20.97) y mujeres (21.54). A modo de conclusión indicar que un mayor nivel de engagement se traduce en un menor nivel de estrés percibido. (AU)

The aim of the study was to determine the influence of leadership, engagement and perceived stress on workers in fitness centres. Conducting a quantitative, descriptive cross-sectional study. The sample consisted of 106 workers, with an average age of 35 years (35.2 ± 8.5). The instruments used were, Multifactor Leadership Questionnaire (MLQ-6S) to measure the type of leadership; the Utrecht Work Engagement Scale (UWES) to know the level of engagement and Perceibed Stress Scale (PSS) to know the perception of stress. Variables, t-Student, ANOVA and Pearson (r) linear correlation and multiple linear regression are compared and correlated. The main results highlight that the level of perceived stress is higher in permanent workers (20.97) and women (21.54). In conclusion, a higher level of engagement translates into a lower level of perceived stress. (AU)

Determinantes socioeconómicos y sociodemográficos asociados al consumo de frutas y verduras de las madres de familia y los hogares de escolares de Jalisco / Socioeconomic and sociodemographic determinants associated with fruit and vegetable consumption among mothers and homes of schoolchildren in Jalisco

Introducción: incrementar el consumo de frutas y verduras es una prioridad, objetivo fundamental en las políticas públicas a nivel mundial. Dado que dicho consumo en los escolares de Jalisco (México) se encuentra por debajo de las recomendaciones alimentarias, es esencial identificar los determinantes que influyen en su consumo de frutas y verduras, para promover el desarrollo de acciones que contribuyan a mejorarlo. Objetivo: identificar los determinantes socioeconómicos y sociodemográficos del consumo de frutas y verduras en las madres de familia y los hogares de escolares de Jalisco, México. Método: estudio transversal analítico, realizado durante el primer trimestre del año 2020 utilizando una frecuencia de consumo de alimentos validada y un cuestionario sobre factores sociodemográficos y socioeconómicos. Resultados: el bajo nivel educativo de las madres de los escolares se asoció con un menor consumo de verduras y el bajo nivel de ingresos en el hogar se asoció con un menor consumo de frutas por los escolares. Conclusiones: el bajo nivel educativo de las madres y el bajo nivel de ingresos de los hogares fueron determinantes asociados al consumo de frutas y verduras de los escolares. Sin embargo, se presentaron diferencias en cuanto a los determinantes para las frutas y para las verduras. Estos factores y sus diferencias deben de tenerse en cuenta para la planificación de acciones que contribuyan a mejorar la ingesta de frutas y verduras en la población escolar (AU)

Introduction: increasing fruit and vegetable consumption is a priority. It has been prioritized as a fundamental objective of public policies worldwide. Given that such consumption in schoolchildren in Jalisco (Mexico) is below the dietary recommendations it is crucial to identify the determinants that influence this consumption to promote the development of contextualized actions that improve it. Objective: to identify the socioeconomic and sociodemographic determinants of fruit and vegetable consumption among mothers and households of schoolchildren in Jalisco, Mexico. Method: an analytical, cross-sectional study carried out during the first quarter of 2020. A validated food consumption frequency and a questionnaire on sociodemographic and socioeconomic factors were used for its development. Results: a lower educational level of the mothers of schoolchildren was associated with a lower consumption of vegetables by schoolchildren. In turn, a lower household income level was associated with a lower consumption of fruits in schoolchildren. Conclusions: a low educational level of mothers and a low household income were determinants associated with fruit and vegetable consumption in schoolchildren. However, there were differences in the determinants for fruits and vegetables. It is essential to consider these factors and their differences in order to plan actions that contribute to improving fruit and vegetable intake in the school population (AU)

Evaluation of a Cooperia oncophora double-domain ASP-based vaccine against Cooperia spp. infections in cattle and sheep.

A double-domain activation-associated secreted protein (dd-Co-ASP) isolated from the bovine small intestinal parasite Cooperia oncophora was previously shown to be an effective vaccine candidate to protect calves against a homologous challenge infection. The aim of this study was to investigate whether the dd-Co-ASP protein, purified from a Belgian C. oncophora isolate, would offer protection against a C. oncophora isolate from the southern hemisphere as well as other Cooperia species such as C. punctata in cattle and C. curticei in sheep. Two vaccination studies were performed, i.e. one in cattle and one in sheep, in which the protective effects of dd-Co-ASP, supplemented with Quil A as an adjuvant, were compared with an adjuvant control. Whereas our results showed a 75 % reduction in Cooperia spp. cumulative faecal egg counts, the results obtained in sheep demonstrated that dd-Co-ASP was ineffective in raising a protective immune response against a C. curticei challenge infection. Even though sequence analysis of the dd-Co-ASP gene revealed restricted sequence heterogeneity in the double domain ASP within and between bovine Cooperia species, the results of the vaccine study suggest that there is sufficient conservation at the protein level to yield cross-protection, holding promise for the development of a general Cooperia vaccine for use in cattle.

Microscopical Techniques to Analyze the Hepatic and Peritoneal Changes Caused by Fasciola hepatica Infection.

The helminth parasite Fasciola hepatica modulates the host immune response at early stages of infection (Rodríguez et al., PLoS Negl Trop Dis 9:e0004234, 2015; Vukman et al., J Immunol 190:2873-2879, 2013). Nevertheless, little is known about the cell composition of the peritoneal fluid at these early stages of infection.In this chapter, we describe a method to perform peritoneal lavages and to recover peritoneal fluid from sheep experimentally infected and noninfected with F. hepatica at early stages of infection. In addition, with the aim to characterize the peritoneal fluid immune cell phenotype, we describe a procedure to obtain the total leukocyte count, the differential leukocyte count and the preparation and storage of peritoneal fluid smears, together with the application of an immunocytochemical technique and an automatic method to count the immunoreactive cells. Finally, the present protocol describes the evaluation of the gross and the histopathological lesions together with the immunohistochemical analysis of the hepatic tissue.

Identification of reference genes for real-time PCR cytokine gene expression studies in sheep experimentally infected with Fasciola hepatica.

The aim of this study was to validate reference genes for gene normalisation using qRT-PCR in hepatic lymph nodes (HLN) and livers from sheep infected with Fasciola hepatica during early and late stages of infection. To this end, a comprehensive statistical approach (RefFinder) encompassing four different methods of analysis (geNorm, BestKeeper, ΔCt method and NormFinder) was used to validate ten candidate reference genes. Stability analysis of gene expression followed by pairwise variation (Vn/Vn + 1) analysis revealed that PGK1, HSP90AA1 and GYPC were the most stable reference genes and suitable for qRT-PCR normalisation in both HLN and liver tissues. These three genes were validated against FoxP3, IL-10, TGF-ß, TNF-α and IL-1ß genes in the HLN tissue of sheep vaccinated with Cathepsin L1 from F. hepatica and unvaccinated infected and uninfected controls during early stages of infection. In the liver, the three reference genes were validated against TNF-α and IL-1ß during chronic stages of infection with F. hepatica and in uninfected controls. Our study is the first to evaluate and validate sheep reference genes in order to provide tools for monitoring cytokines in Fasciola hepatica infected sheep target organs. Our results present an approach to elucidate the role of different cytokines in F. hepatica vaccinated and infected sheep.

Pathological, immunological and parasitological study of sheep vaccinated with the recombinant protein 14-3-3z and experimentally infected with Fasciola hepatica.

In this study, the immunogenicity and protective capacity of a new recombinant vaccine candidate, the rFh14-3-3z protein was analysed in sheep experimentally challenged with Fasciola hepatica, in terms of fluke burden, faecal egg counts, hepatic damage and humoral immune response. Three groups of 8 animals each were used for study, group 1 was immunised with the rFh14-3-3z in Montanide adjuvant, whereas group 2 and 3 remained as adjuvant control and infection control groups, respectively. The parasitological analysis showed that no significant reduction in fluke burden, fluke size and faecal egg counts was detected. The extent of hepatic damage was very similar between groups. Nonetheless, animals immunised with the rFh14-3-3z protein induced the development of specific IgG1 and IgG2, being the IgG1 the predominant antibody; which confirms the immunogenicity of this protein in sheep. This is the first report of the 14-3-3z proteins as vaccine against the infection with F. hepatica.

Study of peritoneal macrophage immunophenotype in sheep experimentally infected with Fasciola hepatica.

During Fasciola hepatica infection, the parasite has the capability to modulate the host immune response towards a non-protector Th2 type instead of Th1. This type of immune response is closely related to the alternative activation of macrophages (M2 profile) as has been shown in vivo in murine models. In this study, an experiment was carried out in order to evaluate the expression of CD68, CD14, CD206 and iNOS in cells present in the peritoneal fluid of sheep during early stages of infection with F. hepatica (1, 3, 9 and 18 days post-infection, dpi) by immunocytochemistry. To the authors' knowledge, this is the first report that studies the in vivo immunophenotype of macrophages from the peritoneal fluid of sheep infected with F. hepatica. Throughout the experiments the absolute number of leucocytes progressively increased, reaching its highest value at 18 dpi, mainly due to the increase of eosinophils. This immunocytochemical study had two purposes: 1) CD68 expression was assessed with Hansel counterstaining, to optimally identify peritoneal macrophages, eosinophils and lymphocytes; 2) expression of CD14, CD206 and iNOS was evaluated to identify alternative or classical pathways of macrophage activation. The results showed a significant increase in CD14 from day 3 dpi compared with the non-infected group. CD206 expression at all time-points showed a significant and dramatic increase in comparison with the uninfected group. On the other hand, iNOS expression showed little variation, and was significantly decreased at 18 dpi in comparison with the uninfected group. These results suggest that F. hepatica induces an alternative activation of peritoneal macrophages of sheep from the first day post-infection, which may facilitate parasite survival. This is the first report describing M2 activation of peritoneal macrophages in ruminants infected with F. hepatica.

Fasciola and fasciolosis in ruminants in Europe: Identifying research needs.

Fasciola hepatica is a trematode parasite with a global distribution, which is responsible for considerable disease and production losses in a range of food producing species. It is also identified by WHO as a re-emerging neglected tropical disease associated with endemic and epidemic outbreaks of disease in human populations. In Europe, F. hepatica is mostly associated with disease in sheep, cattle and goats. This study reviews the most recent advances in our understanding of the transmission, diagnosis, epidemiology and the economic impact of fasciolosis. We also focus on the impact of the spread of resistance to anthelmintics used to control F. hepatica and consider how vaccines might be developed and applied in the context of the immune-modulation driven by the parasite. Several major research gaps are identified which, when addressed, will contribute to providing focussed and where possible, bespoke, advice for farmers on how to integrate stock management and diagnosis with vaccination and/or targeted treatment to more effectively control the parasite in the face of increasing the prevalence of infection and spread of anthelmintic resistance that are likely to be exacerbated by climate change.

Th1/Th2 balance in the liver and hepatic lymph nodes of vaccinated and unvaccinated sheep during acute stages of infection with Fasciola hepatica.

The expression of IFNγ and IL4 was quantified using q-PCR in the liver and hepatic lymph nodes (HLN) of sheep during early stages of infection with Fasciola hepatica (1, 3, 9 and 18days post-infection, dpi). A group of animals (Group 1) were vaccinated with Fasciola hepatica recombinant cathepsin L1 (FhCL1) in montanide 70 VG prior to infection, a second group (group 2) was used as infected control and a third (group 3) was used as uninfected control. To study vaccine efficacy three additional groups were sacrificed 19 weeks post-infection (group 4 immunized with CL1, group 5 with the adjuvant and group 6 was used as infected control). The vaccinated group did not show significant fluke reduction compared to the adjuvant group and infected control group. IL4 expression was observed to increase at 9 dpi and was further elevated at 18 dpi in the liver and HLN of vaccinated and infected control groups compared to the uninfected group. IFNγ expression exhibited different dynamics in the liver and HLN compared to IL4; thus, in the liver this cytokine increased at 9 dpi in the vaccinated and at 18 dpi in vaccinated and infected control groups, while in the HLN it decreased gradually and significantly from 1 dpi onwards. These results suggest that a marked Th2 polarization is present from 9 dpi in HLN and from 18 dpi in the liver. The increase of IFNγ in the liver may correspond with tissue damage response with granuloma formation. The FhCL1 vaccine did not alter the Th1/Th2 balance when compared to unvaccinated and infected sheep. The study of IFNγ and IL4 in the various tissue compartments in sheep could facilitate selection of new adjuvants inducing a strong Th1 response for a more rationale vaccine formulation.

Apoptosis of peritoneal leucocytes during early stages of Fasciola hepatica infections in sheep.

Several immunomodulatory properties have been described in Fasciola hepatica infections. Apoptosis has been shown to be an effective mechanism to avoid the immune response in helminth infections. The aim of the present work was to study apoptosis in peritoneal leucocytes of sheep experimentally infected with F. hepatica during the early stages of infection. Five groups (n=5) of sheep were used. Groups 2-5 were orally infected with 200 metacercariae (mc) and sacrificed at 1, 3, 9 and 18days post-infection (dpi), respectively. Group 1 was used as the uninfected control (UC). Apoptosis was detected using three different methods 1) immunocytochemistry (ICC) with a polyclonal antibody anti-active caspase-3; 2) an annexin V flow cytometry assay using the Annexin V-FITC/propidium iodide (PI); and 3) transmission electron microscopy (TEM). The differential leucocyte count revealed that the majority of peritoneal granulocytes were eosinophils, which increased significantly at 9 and 18 dpi with respect to the uninfected controls. The ICC study revealed that the percentage of caspase-3+ apoptotic peritoneal leucocytes increased significantly from 3 dpi onwards with respect to the uninfected controls. The flow cytometry annexin V assay detected a very significant (P<0.001) increase of apoptotic peritoneal macrophages, lymphocytes and granulocytes, which remained higher than in the UC until 18 dpi. Transmission electron microscopy studies also confirmed the presence of apoptosis in peritoneal eosinophils at 18 dpi. This is the first report of apoptosis induced by F. hepatica in the peritoneal leucocytes of sheep in vivo. The results of this work suggest the importance of apoptosis induction for the survival of the juvenile parasites in the peritoneal migratory stages of infection.

Calidad en el deporte de élite. Análisis de fortalezas y debilidades psicológicas en jugadores de balonmano / Qualidade no esporte de elite. Análise psicológica dos pontos fortes e fracos em jogadores de handebol / Quality in elite sport. Analysis of strengths and weaknesses in psychological handball players

Determinar el perfil psicológico a través de la baremación por modalidades deportivas, en este caso jugadores de balonmano, permite concretar mejor las habilidades psicológicas de cada deporte, ya que cada uno demanda unas diferentes. Por lo que el objeto de estudio es conocer la influencia de las habilidades psicológicas en la ejecución deportiva en jugadores de balonmano categoría élite y determinar los puntos fuertes/débiles psicológicos. Aplicando el Inventario Psicológico de Ejecución Deportiva en una muestra (n=484) con una edad que osciló entre los 12-16 años con un promedio de 13,8 años (dt=1,0) de los cuales el 47,5% eran chicos y el 52,5 chicas. Se realiza un análisis descriptivo para variables cualitativas, frecuencias absolutas y relativas, además de valores mínimos, máximos, media y desviación típica. Se utiliza el coeficiente de relación de Pearson para conocer las correlaciones entre variables. Para determinar las correlaciones del Inventario según género, edad y años de práctica, se hizo la prueba MANOVA. Los resultados destacan que los jugadores de categoría élite de balonmano del estudio tienen un alto control en: afrontamiento positivo, control viso-imaginativo y control actitudinal, mientras que es bajo su control atencional (AU)

Determine the psychological profile through baremación for different sports, in this case handball players, can better realize the psychological skills of each sport, as each demands a different kind. So the object of study is to determine the influence of psychological skills in sports performance in elite handball players category and identify strengths / weaknesses psychological. Applying the Psychological Inventory Sport Running in a sample (n = 484) with an age ranged from 12-16 years with an average of 13.8 years (SD = 1.0), of which 47.5% were boys and girls 52.5. a media descriptive analysis for qualitative variables, absolute and relative frequencies, plus minimum, maximum, and standard deviation is performed. the Pearson correlation coefficient is used to find correlations between variables. To determine the correlations Inventory as gender, age and years of practice, MANOVA was made. the results highlight that the players of handball elite category of the study have a high control: Positive coping imaginative visual-control and attitudinal control while it is under their attentional control (AU)

Determinar o perfil psicológico através baremación para diferentes esportes, neste caso, jogadores de handebol, pode perceber melhor as habilidades psicológicas de cada esporte, como cada um exige um tipo diferente. Assim, o objeto de estudo é determinar a influência das habilidades psicológicas no desempenho desportivo na elite categoria jogadores de handebol e identificar pontos fortes / fracos psicológico. Aplicando o Psychological Inventory do esporte que funciona em uma amostra (n = 484) com uma idade variou de 12-16 anos, com uma média de 13,8 anos (DP = 1,0), dos quais 47,5% eram meninos e meninas 52,5. uma análise descritiva de mídia para as variáveis qualitativas, as frequências absolutas e relativas, além de mínimo, máximo e desvio-padrão é executada. O coeficiente de correlação de Pearson é usado para encontrar correlações entre variáveis. Para determinar o inventário de correlações como sexo, idade e anos de prática, MANOVA foi feita. Os resultados evidenciam que os jogadores do handebol categoria elite do estudo têm um alto controle: lidar positiva imaginativa visual-controle e controlo de atitude enquanto estiver sob o seu controle da atenção (AU)

Distribution of Foxp3+ T cells in the liver and hepatic lymph nodes of goats and sheep experimentally infected with Fasciola hepatica.

Foxp3 regulatory T cells (Tregs) are now considered to play a key role in modulation of immune responses during parasitic helminth infections. Immunomodulation is a key factor in Fasciola hepatica infection; however, the distribution and role of Foxp3+ Tregs cells have not been investigated in F. hepatica infected ruminants. The aim of this study was to evaluate the presence of Foxp3+ Tregs in the liver and hepatic lymph nodes from experimentally infected sheep and goats during acute and chronic stages of infection. Three groups of goats (n=6) and three groups of sheep (n=6) were used in this study. Goats in groups 1-2 and sheep in groups 4-5 were orally infected with metacercarie of ovine origin. Groups 1 and 4 were killed during the acute stage of the infection, at nine days post infection (dpi); groups 2 and 5 were killed during the chronic stage, at 15 and19 weeks post infection respectively (wpi). Groups 3 (goats) and 6 (sheep) were left as uninfected controls. Fluke burdens and liver damage were assessed and the avidin-biotin-complex method was used for the immunohistochemical study. At nine dpi in acute hepatic lesions, the number of both Foxp3+ and CD3+ T lymphocytes increased significantly in goats and sheep. In the chronic stages of infection (15-19wpi), the number of Foxp3+ and CD3+ T lymphocytes were also significantly increased with respect to control livers, particularly in portal spaces with severely enlarged bile ducts (response to adult flukes) while the increase was lower in granulomas, chronic tracts and smaller portal spaces (response to tissue damage). Foxp3+ Tregs were increased in the cortex of hepatic lymph nodes of sheep (chronic infection) and goats (acute and chronic infection). The estimated proportion of T cells which were Foxp3+ was significantly increased in the large bile ducts and hepatic lymph node cortex of chronically infected goats but not sheep. This first report of the expansion of Foxp3+ Tregs in acute and chronic hepatic lesions in ruminants suggests that these cells may be involved in both parasite survival and modulation of hepatic damage. Future studies should be focused on the investigation of parasite molecules and cytokines involved in this process.

Fasciola hepatica induces eosinophil apoptosis in the migratory and biliary stages of infection in sheep.

The aim of the present work was to evaluate the number of apoptotic eosinophils in the livers of sheep experimentally infected with Fasciola hepatica during the migratory and biliary stages of infection. Four groups (n=5) of sheep were used; groups 1-3 were orally infected with 200 metacercariae (mc) and sacrificed at 8 and 28 days post-infection (dpi), and 17 weeks post-infection (wpi), respectively. Group 4 was used as an uninfected control. Apoptosis was detected using immunohistochemistry with a polyclonal antibody against anti-active caspase-3, and transmission electron microscopy (TEM). Eosinophils were identified using the Hansel stain in serial sections for caspase-3, and by ultrastructural features using TEM. At 8 and 28 dpi, numerous caspase-3(+) eosinophils were mainly found at the periphery of acute hepatic necrotic foci. The percentage of caspase -3(+) apoptotic eosinophils in the periphery of necrotic foci was high (46.1-53.9) at 8 and 28 dpi, respectively, and decreased in granulomas found at 28 dpi (6%). Transmission electron microscopy confirmed the presence of apoptotic eosinophils in hepatic lesions at 8 and 28 dpi. At 17 wpi, apoptotic eosinophils were detected in the infiltrate surrounding some enlarged bile ducts containing adult flukes. This is the first report of apoptosis induced by F. hepatica in sheep and the first study reporting apoptosis in eosinophils in hepatic inflammatory infiltrates in vivo. The high number of apoptotic eosinophils in acute necrotic tracts during the migratory and biliary stages of infection suggests that eosinophil apoptosis may play a role in F. hepatica survival during different stages of infection.

Peripheral blood lymphocyte subsets in Fasciola hepatica infected and immunised goats.

The proportions of CD4(+), CD8(+) and WC1+ T lymphocytes from peripheral blood using flow cytometry were investigated in goats infected with Fasciola hepatica and previously immunised with recombinant Cathepsin-L1 (rCL1) and Glutathione-S-transferase sigma class (GST). The immunisation trial did not induce protective responses, and no significant differences were recorded between immunised and non-immunised groups. However, there was a significant decrease in the proportion of CD4(+) T lymphocytes in the infected groups both at 5 weeks post-infection (wpi), coinciding with the migratory stage of the infection, and at 12 wpi in the biliary stage of the infection. The proportional decrease in this circulating population may be related to the recruitment of CD4(+) T cells in liver and hepatic lymph nodes and also to the immunomodulatory effect of the parasite through the interaction of F. hepatica excretory-secretory products (FhESP) with this cell population. To date, this is the first report about the effect of F. hepatica infection in peripheral lymphocyte subsets in goats.

Early hepatic and peritoneal changes and immune response in goats vaccinated with a recombinant glutathione transferase sigma class and challenged with Fasciola hepatica.

Changes and local immune response were evaluated in the peritoneal cell populations, duodenal lamina propria and liver from goats immunized with recombinant glutathione transferase sigma class (rFhGST-S1) during early stages of infection with Fasciola hepatica. Group 1 (n=7) was unimmunized and uninfected; group 2 (n=10) was immunized with adjuvant Quil A and infected; group 3 (n=10) was immunised with rFhGST-S1 and infected. Three goats from each group were killed at 7-9 days post-infection (dpi) to evaluate early changes and immune response. The remaining goats were killed at 15 weeks post-infection (wpi). rFhGST-S1 vaccination induced variable response: three goats showed low fluke burden at 15 wpi and two goats showed low hepatic damage at early infection stages. This response was associated to a severe infiltrate of eosinophils in peritoneal fluid and hepatic necrotic foci, high iNOS expression in peritoneal cells and abundant infiltrate of eosinophils surrounding hepatic migrating flukes. T lymphocyte subsets were found in the vicinity of necrotic areas but they were absent in the vicinity of migrating larvae. No significant variation for T cell subsets, except for CD4 and γδ T lymphocytes, that were higher in the Quil A group compared to the rFhGST-S1 group. Expression of IL4 and IFN-γ in the hepatic inflammatory infiltrates was very occasional.

Early and late peritoneal and hepatic changes in goats immunized with recombinant cathepsin L1 and infected with Fasciola hepatica.

The aim of the present study was to study peritoneal and hepatic changes during early [7-9 days postinfection (dpi)] and late [15 weeks postinfection (wpi)] infection of goats immunized with recombinant F. hepatica pro cathepsin L1 (rCL1) in Quil A and challenged with Fasciola hepatica. Despite finding no significant reduction in fluke burdens between the control and immunized group, at 15 dpi the rCL1-vaccinated group showed significantly higher weight gain and reduced severity of hepatic lesions compared with the control group that received only Quil A. In the rCL1-vaccinated group, two of three goats sacrificed at 7-9 dpi had little hepatic damage and had a higher percentage of peritoneal eosinophils and elevated induced nitric oxide synthase (iNOS) expression in peritoneal cells than the goats from the control group. Moreover, while these two goats showed a heavy infiltration of eosinophils surrounding migrating flukes, the remaining animals examined at 7-9 dpi had no inflammatory infiltration surrounding migrating flukes. Two out of seven goats in the rCL1-vaccinated group had low fluke burdens and little hepatic damage at 15 wpi, suggesting an effective protective response in some of the vaccinated goats. This protective response did not correlate with peripheral eosinophilia or with serum titres of anti-rCL1 immunoglobulin (Ig) G. The results of the present work suggest that an eosinophil-mediated immune response plays a crucial role in the early effective host response against F. hepatica in goats. Adjuvants designed to increase cell-mediated immunity should be tested in future vaccine trials against F. hepatica.

Toxoplasma gondii in ruminant species (cattle, sheep, and goats) from southern Spain.

Seroprevalences of Toxoplasma gondii antibodies were assessed with the use of a commercial indirect ELISA in 1,501 domestic ruminants in southern Spain. Antibodies against T. gondii were detected in 420 (83.3%) of 504 cattle, 248 (49.3%) of 503 sheep, and 124 (25.1%) of 494 goats. The herd seroprevalence was 100% (72/72), 84.7% (61/72), and 72.2% (52/72) for cattle, sheep, and goats, respectively. Seropositivity was significantly higher in herds with a low density of animals (P < 0.001). Significant differences (P < 0.05) among municipalities were also found. The seroprevalence observed in the present study indicates a widespread exposure to T. gondii in livestock in southern Spain.