Graciela Santillán

Immunodiagnosis of human fascioliasis by an enzyme-linked immunosorbent assay (ELISA) and a micro-ELISA.

ABSTRACT Enzyme-linked immunosorbent assay (ELISA) and micro-ELISA were evaluated for their ability to detect anti-Fasciola hepatica antibodies in humans by using excretory-secretory antigen. The sensitivity of each method was 100%, but the specificity was 100% for ELISA and 97% for micro-ELISA. The micro-ELISA could be used as a screening assay and ELISA could be used as a confirmatory method for the serodiagnosis of human fascioliasis.

SEROEPIDEMIOLOGY OF HUMAN HYDATIDOSIS: USE OF DRIED BLOOD SAMPLES ON FILTER PAPER

Programmes for the control of hydatidosis caused by Echinococcus granulosus in Argentina use an enzyme immunoassay (EIA) as a screening test for population surveys aimed at detecting asymptomatic patients. Persons thus selected are referred to health centres for the arc 5 double diffusion test and imaging techniques. One of the most costly procedures of these surveys is the collection of blood samples under field conditions; the possibility of collecting dry blood samples on filter paper was therefore investigated. In a survey of 497 rural inhabitants of an endemic area, the same number of hydatidosis cases (22) were identified by EIA using (i) serum samples and (ii) capillary blood samples obtained by finger prick and collected on filter paper. The latter system was both simpler and cheaper.

Pilot field trial of the EG95 vaccine against ovine cystic echinococcosis in Rio Negro, Argentina: Early impact and preliminary data

Cystic echinococcosis is endemic in the Rio Negro province of Argentina. After 30 years of control using praziquantel in dogs the transmission rate to humans and sheep has decreased significantly, however transmission persists. The objective of the study is to assess the impact of the inclusion of the EG95 vaccine for sheep in the control programme, including analysis of the vaccines operative feasibility in field conditions. The vaccine was applied in an area comprising four communities of native people including 79 farms with 3146 lambs and 311 dogs in total. Seventy one farms were designated as control areas where no vaccinations were undertaken while vaccinations of lambs undertaken on 91 farms. Lambs received two vaccinations with the EG95 vaccine followed by a single booster injection when the animals were 1-1.5 years of age. Farm locations were defined using GPS coordinates for the houses. Evidence for Echinococcus granulosus transmission was monitored by coproantigen ELISA on samples of dog faeces, by E. granulosus-specific PCR using soil samples, and anti-E. granulosus antibody assessments in sera from 2 to 4 teeth lambs, purgation of dogs to detect E. granulosus worms and necropsy on adult sheep. Before the vaccine was introduced, 26.2% of sheep with 2-4 teeth were positive using ELISA/WB, the prevalence decreased to 7.8% at the third year following use of the vaccine. Necropsy of animals older than 6 years (not vaccinated) showed that 66.1% of animals were infected with E. granulosus. In dogs, 4% was found positive for E. granulosus using arecoline purgation and 24.7% of the farms were infected using coproELISA/WB. During the first year of vaccination 2721 lambs received the first vaccine dose and 2448 received a booster. In the second year 2138 lambs were initially vaccinated and 1745 received a booster, and 1308 animals received the third dose. During the third year 1110 lambs received the first dose from which 539 received a booster and 723 animals received the third dose. An analysis of advantages and limitations of the diagnostic techniques used and the ability of the geospatial analysis to detect risk area are included. Based in the immunodiagnostic techniques, the EG95 vaccine has been able to prevent the infection in animals up to 3 years old. Also, the difficulties in the field for the correct vaccine administration and the social features and habits that may impact on echinococcosis control are included in the analysis.

Pilot Field Trial of the EG95 Vaccine Against Ovine Cystic Echinococcosis in Rio Negro, Argentina: Second Study of Impact.

Background Cystic echinococcosis (CE) is an important zoonotic disease caused by the cestode parasite Echinococcus granulosus. It occurs in many parts of the world where pastoral activities predominate, including the Rio Negro province of Argentina. Although CE control activities have been undertaken in the western regions of Rio Negro for more than two decades, the disease continues to remain prevalent in both the human and livestock animal populations. Vaccination of animal intermediate hosts of CE with the EG95 vaccine may provide a new opportunity to improve the effectiveness of CE control measures, although data are lacking about field application of the vaccine. Aims Evaluate the impact of EG95 vaccination in sheep on the transmission of Echinococcus granulosus in a field environment. Methodology Two trial sites were established in western Rio Negro province within indigenous communities. Vaccination of lambs born into one trial site was introduced and continued for 6 years. Prior to initiation of the trial, and at the end of the trial, the prevalence of CE in sheep was determined by necropsy. Weaned lambs received two injections of EG95 vaccine, approximately one month apart, and a single booster injection one year later. Vaccination was not implemented at the second trial site. A total of 2725 animals were vaccinated in the first year. Animals from this cohort as well as age-matched sheep from the control area were evaluated by necropsy. Key results Introduction of the vaccine led to a statistically significant in the number and size of hydatid cysts in comparison to the situation prior to the introduction of the vaccine, or compared to CE prevalence in the control area where the vaccine was not applied. The prevalence of infection in the vaccinated area was also significantly reduced by 62% compared to the re-intervention level, being lower than the prevalence seen in the control area, although the difference from the control area after the intervention was not significant possibly due to limitations in the numbers of animals available for necropsy. Conclusions Vaccination of sheep with the EG95 vaccine provides a valuable new tool which improves the effectiveness of CE control activities. Vaccination was effective even in a difficult, remote environment where only approximately half the lambs born into the communities were fully vaccinated.

Diagnóstico de situación de la equinococosis quística en heces dispersas en las zonas de Quebrada y Puna, provincia de Jujuy, Argentina

Resumen es: La equinococosis es una enfermedad parasitaria comun en el ganado, causada por el ces- tode Echinococcus granulosus; el perro es su principal hospedador ...

Evaluación de un antígeno purificado para el diagnóstico de toxocariosis

La toxocariosis es una zoonosis causada por la ingestion de huevos infectivos de Toxocara spp. El diagnostico de la enfermedad se basa en la deteccion de anticuerpos en el suero u otros fluidos biologicos. La tecnica serologica mas utilizada es el ELISA, que usa como antigeno los productos de excrecion-secrecion de larvas de tercer estadio (ES/L3). Estos productos antigenicos son glicoproteinas que se originan en los organos secretorios del parasito y no son especificos de especie. Para evaluar la especificidad de la tecnica de ELISA con el antigeno ES/L3, se emplearon sueros de personas con otras helmintiasis y con patologias no parasitarias. Se observo que estos sueros presentaron reactividad entre el 11 y el 70 % de los casos. El Western blot con suero de los mismos pacientes revelo que la glicoproteina que corresponde al triplete de 120 kDa fue la mas inespecifica. Teniendo en cuenta estos resultados y con el proposito de purificar el antigeno se realizo una cromatografia de intercambio ionico. Cuando se analizaron los sueros de los pacientes con diferentes enfermedades parasitarias y no parasitarias con el antigeno ES/L3 purificado, solo fueron reactivos entre un 10 y un 20 % de ellos. La sensibilidad del test de ELISA determinada por el programa Epidat 3. 0 para los dos antigenos fue del 100 %, pero se observaron diferencias en la especificidad: para el antigeno ES/L3 total esta fue del 84 % y para el ES/L3 purificado del 99 %. Empleando el antigeno ES/L3 purificado se puede considerar que los sueros que son reactivos, en presencia de una sintomatologia compatible, corresponden a pacientes que fueron o estan parasitados con Toxocara canis.

Detección de coproantígenos para el diagnóstico de echinococosis canina en la zona fronteriza de La Quiaca-Villazón

Cystic Echinococcosis is a major public health issue. Immunodiagnostic techniques based on the identification of parasite antigens in dog dry faeces have been developed as alternatives for the surveillance of canine Echinococcosis. The environmental conditions favouring the parasite cycle were met in the border of La Quiaca-Villazon, given the presence of definitive (dog) and intermediate hosts (sheep and goats). The most important activity in La Puna is sheep and goat rearing; slaughtering is carried out almost exclusively in the field by sheeps owners, and preventive health measures do not apply. The aim of this study was to determine the presence of dogs parasitized by Echinococcus granulosus in this border region. A hundred and sixty eight (168) stool specimens were collected during 2006. La Quiaca samples were taken from the following selected areas: the semi-rural area of Barrios, the rural areas of Santa Catalina, Yavi Chico, El Portillo, Pumahuasi and Cara Cara and the urban area of La Quiaca; selected urban areas in Villazon and the semi-rural area of Ojo de Agua, Lampaya and Matancillas. The samples were analyzed by copro-ELISA -Western blot test. The cities of San Francisco and Barrios had a prevalence of 14.3% and 6.7%, respectively. A prevalence of 3.4% was observed in the urban area of Villazon, which indicates that dogs become infected in the rural areas and bring the risk into the city. Lampaya showed a prevalence of 30%. These findings suggest the need to implement strategies for the control of hydatidosis, both in urban and rural areas to avoid the increase and spread of Echinococcosis in the region.

Pilot field trial of the EG95 vaccine against ovine cystic echinococcosis in Rio Negro, Argentina: Humoral response to the vaccine

Fil: Larrieu, Edmundo Juan. Universidad Nacional de La Pampa. Facultad de Ciencias Veterinarias; Argentina. Universidad Nacional de Rio Negro; Argentina

Vigilancia de la equinococosis quística en perros y niños en la provincia de Río Negro, Argentina

Cystic echinococcosis (CE) is an endemic disease in the province of Río Negro, Argentina. The control program against CE has developed monitoring surveillance systems. Currently, the coproELISA/Western blot (WB) test is used to determine transmission in livestock farms (epidemiological units or EU) from collected dry field-dispersed dog feces. The objective was to evaluate the prevalence of CE on livestock farms and its relationship with CE cases in children aged 0-14 years. Canine fecal samples were obtained from randomly selected livestock farms and processed by the coproELISA/WB test. Furthermore, new cases in children in the same age group mentioned above were identified. In 278 EU, 571 samples of canine feces were obtained. There were 37 positive samples for coproELISA/WB (6.5%) and the presence of transmission was demonstrated in 37 EU (13.3%). There were no significant differences (p=0.9) with the survey conducted in the period 2003-2004 while there were significant differences (p=0.02) with the EU survey of native populations conducted in 2009-2010. With respect to animal density in the work area, the EU yielding negative results had an average of 2 dogs (SD 2.1) per EU while in the EU having positive results the average was 3 dogs (SD 4.2), showing statistically significant differences (p=0.02). In children under 15 years of age, 12 cases were diagnosed. This study has identified, on average, that the cases of hydatid disease are closer in the positive fields than in the negative fields (p=0.00307).The coproELISA/WB test allowed to identify the dispersion of CE on livestock farms and its relationship with the occurrence of cases in children in 2009-2010.Resumen es: La equinococosis quistica (EQ) es una enfermedad endemica en la provincia de Rio Ne- gro, Argentina. El programa de control de esta afeccion desarrolla s...