Control programme for cystic echinococcosis in Uruguay.

Cystic echinococcosis is a highly endemic parasitic zoonosis that is present in the Southern Cone countries of America. For several decades, various prevention and control programmes have been implemented in different countries and regions, with varying results. In Uruguay, a new control programme was implemented in 2006 that employed new strategies for canine diagnosis and treatment, dog population control, diagnosis in humans, epidemiological surveillance, and health education, including community participation. The control programme in Uruguay addresses the control and surveillance of the disease from a holistic perspective based on Primary Health Care, which has strengthened the community's participation in developing and coordinating activities in an interdisciplinary manner. Similarly, the control programme that is currently implemented is based on a risk-focused approach. The surveillance and control measures were focused on small villages and extremely poor urban areas. In this study, the strategies used and the results obtained from 2008-2013 are analysed and discussed.

Profilin cross-reactive panallergen causes latex sensitization in the pediatric population allergic to pollen.

BACKGROUND: Component-resolved diagnostics (CRD) has been demonstrated to be an excellent new tool for improving the current diagnosis of allergies, and it allows differentiation between polysensitization and cross-reactivity. OBJECTIVE: To demonstrate the role of cross-reactive pollen allergens in pediatric patients living in areas with large amounts of airborne grass pollen grains who are sensitive to grass pollen and latex. METHODS: Serum samples were obtained from 106 children between 3 and 14 years of age diagnosed with allergies to pollen based on clinical history, skin prick tests, and specific immunoglobulin E (IgE). None of them had allergy symptoms to latex or fruits. From these 106 children, 56 patients revealed positive results to Phleum-specific major allergens but not to cross-reactive allergens. The other 50 patients who showed positive specific IgE to Phleum-specific major allergens and to cross-reactive pollen allergens also showed positive results to latex allergens. CRD was carried out by specific IgE quantification using a fluoro-enzyme immunoassay (ImmunoCAPT System). RESULTS: Results demonstrated a positive significant relationship between the specific IgE to Hev b 8 and Phl p 12 and also between the specific IgE to Hev b 8 and latex extract in the group of patients sensitized to species-specific and cross-reactive Phleum allergens. Positive significant relationships were also found between profilin and avocado or peach sensitizations. No other latex allergens gave positive results. CONCLUSION: The apparent sensitization to latex in pediatric patients allergic to grass pollen is caused by the cross-reactive profilin panallergen; however, it is appears not to be clinically relevant.

Identification of allergens homologous to Alt a 1 from Stemphylium botryosum and Ulocladium botrytis.

Several studies have demonstrated that proteins homologous to the Alt a 1 major allergen of Alternaria alternata are expressed in other members of the Pleosporaceae family. However, since no direct biochemical data have been reported concerning the presence of Alt a 1 allergen homologues capable of binding IgE in the excretion-secretion products of Stemphylium and Ulocladium, our objective was to explore their presence in Stemphylium botryosum and Ulocladium botrytis. S. botryosum and U. botrytis culture filtrate extracts were analyzed by two-dimensional (2D)-electrophoresis and 2D-immunoblotting using polyclonal rabbit antibodies raised against recombinant Alt a 1, as well as five human sera from patients allergic to Alternaria. Cross-reactivity immunoassays were performed by ImmunoCAP inhibition and 2D-immunoblotting inhibition. IgE-binding proteins recognized by the rabbit antiserum raised against Alt a 1, with apparent molecular weights of 17-18 kDa and isoelectric points of 4, were identified as Alt a 1-like proteins. Alt a 1 inhibited IgE-specific binding to the Alt a 1 homologues from S. botryosum and U. botrytis. In conclusion, it was demonstrated that allergens which are homologous to Alt a 1 are expressed in the excretory-secretory materials of the phylogenetically-related species S. botryosum and U. botrytis.

Dog echinococcosis in northern Spain: comparison of coproantigen and serum antibody assays with coprological exam.

A large sheep-dog population from the province of Alava (northern Spain) has been investigated in order to determine the prevalence of the cestode parasite Echinococcus granulosus. Worms were detected in 14.0% of 721 dog faecal supernatants by coproantigen ELISA, and in 9.1% of 754 dog serum samples by serum antibody ELISA. A weak but statistically significant correlation (Spearman's rho=0.103, 95% CI: 0.023-0.178) between the two immunoassay results was found. In addition, eggs of the family Taeniidae were detected in 10.3% of 726 faecal samples examined by coproparasitological (flotation and sedimentation) tests. The overall E. granulosus infection rate, based on a Bayesian latent class model that accounts for the imperfect sensitivities and specificities of all diagnostic tests used, was estimated to be 8.0% (95% credible interval: 5.4-11.4%), corroborating that sheep-dog is the dog class most vulnerable to acquiring the infection. Dog sex did not influence the prevalence of E. granulosus, independently of the diagnostic test used or the dog region of origin. No significant linear correlation was found between the coproantigen ELISA OD values and the dog age (Spearman's rho=-0.049, 95% CI: -0.234 to 0.135), suggesting that there were no differences in prevalence of E. granulosus between old and young dogs. The obtained results highlight the importance of initiating a control program based on regular treatment of the sheep-dogs with praziquantel in the province of Alava.

El hospital como dinamizador y difusor de salud regional: la experiencia del Hospital de Tacuarembó, Uruguay / Hospitals as a way to dynamize and disemminate regional health: the case of the Hospital de Tacuarembó, Uruguay / O hospital como habilitador e difusor da saúde regional: a experiência do hospital de Tacuarembó, Uruguai

Resumen: Se describe el desarrollo del Hospital Regional de Tacuarembó (HRT), situado en una ciudad mediana del norte de Uruguay, que lo condujo a convertirse en un referente dinamizador de la salud regional, con una atención médica moderna y eficaz, así como de servicio a la comunidad y a la salud nacional. Todo ello logrado a través de una gestión ambiciosa y optimista en los objetivos, basada en esfuerzos colectivos e individuales así como en apoyos institucionales nacionales y de organismos internacionales. Ese papel dinamizador e influyente a nivel regional, e inclusive nacional, está basado, por una parte, en una gran mejora de su base logística, es decir, el propio hospital, en cuanto a instalaciones, equipamientos y ampliación de las capacidades de atención médica; pero también en un gran apoyo e impulso a la Atención Primaria en Salud, no solo en la ciudad de Tacuarembó, sino a nivel departamental. Además, el HRT ha jugado un papel importante en el control de dos zoonosis parasitarias de importancia nacional: la enfermedad de Chagas y la equinococosis quística. Esa acción dinamizadora ha consistido en plantear al hospital como una "organización de salud", considerando el contexto de un territorio particular junto a los determinantes del sistema nacional de salud. Se puede considerar al HRT como una organización innovadora liderada por "emprendedores sociales" dentro de un marco de emprendimiento social partiendo de un hospital departamental o provincial.

Summary: The study presents the case of the Hospital de Tacuarembó (HRT), located in a medium-size city in the north of Uruguay, which turned into a reference in terms of the dynamization of regional health, offering modern and effective healthcare services, serving the community and contributing to the improvement of national health. All of this was possible as a result of an ambitious and optimistic management of objectives, based on collective and individual effort, as well as relying on the support of national and international bodies. This dynamizing and influential role played by the Hospital de Tacuarembó is based, on the one hand on a great improvement of its logistical base - that is, the hospital itself - in terms of its premises, equipment and upscaling of medical services, and on the other hand on the major support and stimulus to Primary Healthcare Services, not only in the city of Tacuarembó, but also throughout the entire province. Likewise, the HRT has played a significant role in the control of two parasite zoonoses that are extremely important for Uruguay: Chagas disease and cystic echinococcosis. This dynamizing actions have consisted in regarding the hospital as a "health organization" within the context of a certain territory, along with factors defined by the national health system. HRT may be considered as an innovative organization led by "social entrepreneurs" within the framework of social entrepreneurship, stemming from a departmental or province hospital.

Resumo: Descreve-se o desenvolvimento do Hospital Regional de Tacuarembó (HRT), localizado em uma cidade de médio porte do norte do Uruguai, o que o tornou uma referência dinâmica de saúde regional, com atendimento médico moderno e eficaz, além de atendimento à comunidade e a saúde nacional. Tudo isto alcançado através de uma gestão ambiciosa e otimista dos objectivos, baseada no esforço coletivo e individual, bem como no apoio institucional nacional e de organismos internacionais. Este papel dinâmico e influente a nível regional, e mesmo nacional, está baseado, por um lado, numa grande melhoria da sua base logística, ou seja, do próprio hospital, ao nível das instalações, equipamentos e ampliação das capacidades de assistência médica; mas também num grande apoio e dinamização da Atenção Primária à Saúde (APS) não só na cidade de Tacuarembó mas também a nível departamental. Além disso, o HRT tem desempenhado um papel importante no controle de duas zoonoses parasitárias de importância nacional: a doença de Chagas e a equinococose cística. Essa dinâmica de ação consistiu em considerar o hospital como uma "organização de saúde", considerando o contexto de um determinado território e os determinantes do sistema nacional de saúde. O HRT pode ser considerado uma organização inovadora liderada por "empreendedores sociais" dentro de um quadro de empreendedorismo social a partir de um hospital departamental ou provincial.

Control programme for cystic echinococcosis in Uruguay

Cystic echinococcosis is a highly endemic parasitic zoonosis that is present in the Southern Cone countries of America. For several decades, various prevention and control programmes have been implemented in different countries and regions, with varying results. In Uruguay, a new control programme was implemented in 2006 that employed new strategies for canine diagnosis and treatment, dog population control, diagnosis in humans, epidemiological surveillance, and health education, including community participation. The control programme in Uruguay addresses the control and surveillance of the disease from a holistic perspective based on Primary Health Care, which has strengthened the community’s participation in developing and coordinating activities in an interdisciplinary manner. Similarly, the control programme that is currently implemented is based on a risk-focused approach. The surveillance and control measures were focused on small villages and extremely poor urban areas. In this study, the strategies used and the results obtained from 2008-2013 are analysed and discussed.

A new application of Streptavidin ImmunoCAP for measuring IgG antibodies against non-available commercial antigens.

BACKGROUND: Enzyme immunoassay has been successfully used for the diagnosis of hypersensitivity pneumonitis (HP), but there are difficulties in standardising this method because of the difficulty in obtaining stable solid phases from a huge variety of heterogeneous antigenic sources. With this in mind, the aim of this work was to demonstrate the usefulness of the standard available fluoro-enzyme-immuno-assay (FEIA) to measure specific IgG to uncommon, commercially unavailable HP-causing antigens. METHODS: Serum samples from patients suffering from HP caused by canary and pigeon protein exposure with positive immunoprecipitation values, as well as cystic fibrosis patients, patients with respiratory allergies, asymptomatic subjects exposed to canary proteins (canary breeders) and healthy unexposed individuals were evaluated by FEIA using biotinylated serum proteins coupled to streptavidin-activated solid phases. Statistical analyses were performed to compare the different groups and to evaluate the performance of this technique. RESULTS: The group of patients with hypersensitivity pneumonitis caused by exposure to canary proteins showed the highest value of specific IgG (35.70 mg IgG/mL). ROC analysis demonstrated an optimal cut-off value of 8.44 mg IgG/mL with a sensitivity of 100% and a specificity of 91.76%. Comparisons with other groups revealed statistically significant differences. Comparison between ELISA and FEIA results revealed a strong positive relationship between values obtained in both assays. CONCLUSION: The results demonstrate that Streptavidin ImmunoCAP is a sensitive, specific and efficient laboratory method for routine diagnostic testing of HP caused by protein antigens that are not commercially available.

Bovine serum albumin contained in culture medium used in artificial insemination is an important anaphylaxis risk factor.

OBJECTIVE: To analyze the cause of the anaphylactic reaction after a standard artificial insemination process in a patient diagnosed with asthma. DESIGN: Case report. SETTING: Residencia Sanitaria Virgen de la Arrixaca (Murcia, Spain) and University of the Basque Country (Vitoria, Spain). PATIENT(S): A 30-year-old woman with a previous medical history compatible with respiratory allergy who suffered an anaphylactic reaction after an artificial insemination with spermatozoids in capable medium (Upgraded B2 INRA medium; Laboratories CCD, Paris, France). INTERVENTION(S): Cutaneous tests and specific IgE levels to inhalant allergens, grass and Olea pollens, and insemination medium were performed. MAIN OUTCOME MEASURE(S): Specific IgE levels to mammal epithelia and bovine serum albumin (BSA). RESULT(S): Skin prick tests were positive for inhalant allergens such as mites, cat, dog, horse, and rabbit epithelia, grasses and Olea pollens, and the insemination medium. The beta-lactamic tests were negative. The determination of specific IgE demonstrated positive values to mammal epithelia and mammal serum albumins including BSA. CONCLUSION(S): We report a case of an anaphylactic reaction to the BSA included in the insemination culture medium induced by a subclinical sensitivity to serum albumins of mammal epithelia. A previous testing with the medium is recommended and specific testing might be needed in women who have a history of animal epithelium allergies.

Enzymatic activities of Alternaria alternata allergenic extracts and its major allergen (Alt a 1).

Several allergens from Alternaria alternata have been isolated allowing some of them to be identified and characterised. Despite the fact that the major allergen of A. alternata (Alt a 1) has been extensively produced by recombinant technology, its biological activity still remains unknown. In the present study, extracts from culture filtrates were used to evaluate the intra-specific variability of the enzymes and also as a source for isolating and purifying native Alt a 1. This was purified by affinity chromatography using antibody anti-recombinant Alt a 1 (produced in Escherichia coli). Enzyme activities were analysed by the API-ZYM System screening method. Results demonstrated the high variability of enzyme activities among the different strains. Only activities corresponding to phosphatases, esterases and beta-glucosidase were expressed by 100% of the strains. Both native and recombinant Alt a 1 showed phosphatase and esterase activities, suggesting that the glucidic moiety of this allergen does not significantly affect its enzyme activity.

Shared and non-shared antigens from three different extracts of the metacestode of Echinococcus granulosus.

Hydatid cyst fluid (HCF), somatic antigens (S-Ag) and excretory-secretory products (ES-Ag) of Echinococcus granulosus protoscoleces are used as the main antigenic sources for immunodiagnosis of human and dog echinococcosis. In order to determine their non-shared as well as their shared antigenic components, these extracts were studied by ELISA-inhibition and immunoblot-inhibition. Assays were carried out using homologous rabbit polyclonal antisera, human sera from individuals with surgically confirmed hydatidosis, and sera from dogs naturally infected with E. granulosus. High levels of cross-reactivity were observed for all antigenic extracts, but especially for ES-Ag and S-Ag. Canine antibodies evidenced lesser avidity for their specific antigens than antibodies from human origin. The major antigenic components shared by HCF, S-Ag, and ES-Ag have apparent molecular masses of 4-6, 20-24, 52, 80, and 100-104 kDa, including doublets of 41/45, 54/57, and 65/68 kDa. Non-shared polypeptides of each antigenic extract of E. granulosus were identified, having apparent masses of 108 and 78 kDa for HCF, of 124, 94, 83, and 75 kDa for S-Ag, and of 89, 66, 42, 39, 37, and 35 kDa for ES-Ag.

Preliminary study of the presence of antibodies against excretory-secretory antigens from protoscoleces of Echinococcus granulosus in dogs with intestinal echinococcosis.

The aim of the present study was to analyze the antibody response against excretory-secretory antigens (ES-Ag) from Echinococcus granulosus protoscoleces, using sera from dogs infected with E. granulosus and other helminths. ES-Ag were obtained from the first 50 h maintenance of protoscoleces in vitro. Immunochemical characterization was performed by immunoblotting with sera from dogs naturally infected with E. granulosus (n = 12), sera from dogs infected with helminths other than E. granulosus (n = 30), and helminth-free dog sera (n = 20). These findings were compared to those obtained from a somatic extract of protoscoleces (S-Ag). ES-Ag only showed four cross-reacting proteins of 65, 61, 54, and 45-46 kDa. Antigens with apparent masses of 89 and 50 kDa in ES-Ag and of 130 and 67 kDa in S-Ag were identified by sera of dogs infected with E. granulosus only, whereas a protein of 41-43 kDa was recognised by the majority of the sera from dogs with non-echinococcal infection. Employing ELISA to study the same sera, S-Ag revealed higher immunoreactivity than ES-Ag, but also showed higher cross-reactivity levels when sera from dogs with non-echinococcal infection were assayed in immunoblotting.

Shared and non-shared antigens from three different extracts of the metacestode of Echinococcus granulosus

Hydatid cyst fluid (HCF), somatic antigens (S-Ag) and excretory-secretory products (ES-Ag) of Echinococcus granulosus protoscoleces are used as the main antigenic sources for immunodiagnosis of human and dog echinococcosis. In order to determine their non-shared as well as their shared antigenic components, these extracts were studied by ELISA-inhibition and immunoblot-inhibition. Assays were carried out using homologous rabbit polyclonal antisera, human sera from individuals with surgically confirmed hydatidosis, and sera from dogs naturally infected with E. granulosus. High levels of cross-reactivity were observed for all antigenic extracts, but especially for ES-Ag and S-Ag. Canine antibodies evidenced lesser avidity for their specific antigens than antibodies from human origin. The major antigenic components shared by HCF, S-Ag, and ES-Ag have apparent molecular masses of 4-6, 20-24, 52, 80, and 100-104 kDa, including doublets of 41/45, 54/57, and 65/68 kDa. Non-shared polypeptides of each antigenic extract of E. granulosus were identified, having apparent masses of 108 and 78 kDa for HCF, of 124, 94, 83, and 75 kDa for S-Ag, and of 89, 66, 42, 39, 37, and 35 kDa for ES-Ag.

Preliminary study of the presence of antibodies against excretory-secretory antigens from protoscoleces of Echinococcus granulosus in dogs with intestinal echinococcosis

The aim of the present study was to analyze the antibody response against excretory-secretory antigens (ES-Ag) from Echinococcus granulosus protoscoleces, using sera from dogs infected with E. granulosus and other helminths. ES-Ag were obtained from the first 50 h maintenance of protoscoleces in vitro. Immunochemical characterization was performed by immunoblotting with sera from dogs naturally infected with E. granulosus (n = 12), sera from dogs infected with helminths other than E. granulosus (n = 30), and helminth-free dog sera (n = 20). These findings were compared to those obtained from a somatic extract of protoscoleces (S-Ag). ES-Ag only showed four cross-reacting proteins of 65, 61, 54, and 45-46 kDa. Antigens with apparent masses of 89 and 50 kDa in ES-Ag and of 130 and 67 kDa in S-Ag were identified by sera of dogs infected with E. granulosus only, whereas a protein of 41-43 kDa was recognised by the majority of the sera from dogs with non-echinococcal infection. Employing ELISA to study the same sera, S-Ag revealed higher immunoreactivity than ES-Ag, but also showed higher cross-reactivity levels when sera from dogs with non-echinococcal infection were assayed in immunoblotting.