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Cystic echinococcosis (CE) is a zoonosis caused by species of the complex Echinococcus granulosus, sensu lato in their larval stage. It is an endemic disease in the province of Río Negro, where small farmers generally have both sheep and goats. Lamb vaccination with EG95 was incorporated in 2009 with very good results: in fact, it contributed to a significant drop in prevalence of infection in both sheep and goats, when determined by necropsy and serology in 2018. In the design of the activity, it was decided not to vaccinate goats in order to minimize the operational requirements of vaccination and comments from producers about the rarity of observing hydatid cysts in goat viscera were considered.To identify causes which can still generate infection in dogs, and to detect species/genotypes in circulation in the province of Río Negro.In indigenous reserves comprised within the area of lamb vaccination with 3 doses of EG95, (dose 1 in December, dose 2 in January and dose 3 in December of the year following, at the time of application of dose 1 to the new lambs). Prevalence in adult goats and sheep was determined by necropsy and serology (ELISA). Infective species/genotypes present in the work area and in the rest of the province of Río Negro were identified by Cox1 mitochondrial gene sequencing. Epidemiological analysis was completed with surveys among farmers about slaughter habits for human consumption.Through serology and necropsy, infection rates in vaccinated and nonvaccinated sheep were significantly different (21% versus 66%). Non-vaccinated sheep and non-vaccinated goats were also significantly different in that there was less infection in goats compared to sheep (7% versus 66% for necropsy, 30% versus 61% for serology); After many years of sheep vaccination the infection positives were low, and differences between vaccinated sheep and non-vaccinated goats turned out non-significant (21% versus 7%). With reference to epidemiology and control along the period 2018-2022, PZQ dosing of dogs 4 times a year was maintained, and 2 extra deworming tasks were introduced together with dose 1 and 2 of EG95, performed by the veterinary vaccination team, ensuring the ingestion of PZQ by dogs. Assessment of animal slaughter for consumption in 41 producers showed that 21 of them slaughter a monthly average of 18 goats (an average of 0.43 goat per month per farm) and 36 in all slaughter 35 old sheep in a year (average of 0.85 sheep per month per farm). With respect to identification of species/genotypes as from 2010, genotypes G1 have been found in 11 sheep (out of which 6 belong to vaccination zone) and genotypes G7, in one pig. A goat cyst within vaccination zone turned out unfertile and it was not possible to sequence it.Design and implementation of a vaccine programme combined with the use of PZQ resulted as cost-effective, since it was possible to maintain the vaccine over time, with clear impact on prevalence decrease in sheep and goats.

Background Cystic echinococcosis (CE) is a parasitic zoonosis caused by infection with the larval stage of Echinococcus granulosus (s.l.). This study investigated the prevalence and potential risk factors associated with human CE in the towns and rural areas of Ñorquinco and Ramos Mexia, Rio Negro province, Argentina. Methods To detect abdominal CE cysts, we screened 892 volunteers by ultrasound and investigated potential risk factors for CE using a standardized questionnaire. Prevalence ratio (PR) with 95% confidence intervals (CI) was used to measure the association between CE and the factors investigated, applying bivariate and multivariate analyses. Results Abdominal CE was detected in 42/892 screened volunteers (4.7%, 95% CI 3.2–6.1), only two of whom were under 15 years of age. Thirteen (30.9%) CE cases had 25 cysts in active stages (CE1, CE2, CE3a, according to the WHO Informal Working Group on Echinococcosis [WHO-IWGE] classification). The most relevant risk factors identified in the bivariate analysis included: living in rural areas (P = 0.003), age > 40 years (P = 0.000), always drinking water from natural sources (P = 0.007), residing in rural areas during the first 5 years of life (P = 0.000) and having lived more than 20 years at the current address (P = 0.013). In the multivariate final model, the statistically significant risk factors were: frequently touching dogs (P = 0.012), residing in rural areas during the first 5 years of life (P = 0.004), smoking (P = 0.000), age > 60 years (P = 0.002) and living in rural areas (P = 0.017). Conclusions Our results point toward infection with CE being acquired since childhood and with constant exposure throughout life, especially in rural areas with a general environmental contamination. Graphical Abstract

Cystic echinococcosis (CE) 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 was to assess the inclusion of the EG95 for sheep in the control program and to determine the vaccine's operative feasibility in field conditions. An intervention study was defined in Rio Negro Province in Argentina comprising, in total, an area of 5820 Km2. Lambs received two vaccinations with the EG95 vaccine followed by a single booster injection when the animals were 1-1.5 years of age. Vaccination of lambs born into one trial site was introduced and continued for 8 years. Evidence for Echinococcus granulosus transmission was monitored before and after vaccination by coproantigen ELISA in faecal samples of dog, purgation of dogs to detect E. granulosus worms, necropsy on adult sheep and by ultrasound screening in children of 6-14 years old. 29,323 doses of vaccine were applied between 2009 and 2017, which a vaccination coverage of 80.1%/85.7% (57.3% average for fully vaccinated). Before the introduction of the vaccine 56.3% of the 6-year-old sheep were infected with E. granulosus at necropsy and 84.2% of the farms had infected sheep; 4.3% of the dogs were positive for E. granulosus infection using the arecoline test, and with coproELISA 9.6% of dog fecal samples were positive and 20.3% of the farms had infected dog.After the vaccine was introduced, 21.6% of sheep older than 6 years were found to be infected at necropsy and 20.2% of the farms were found to be infected; in dogs, 4.5% were found positive for E. granulosus using arecoline purgation and with coproELISA 3.7% of samples were positive, with 8.9% of farms having a positive dog. In 2016 only one case of E. granulosus infection was diagnosed by US screening in a 6-14 years old child. Included in the analysis are discussions of difficulties experienced in the field which affected correct vaccine administration as well as social features and practices that may impact on echinococcosis control and the EG95 vaccination program in Rio Negro. 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.

Background Rio Negro Province is endemic for cystic echinococcosis (CE). A CE control program includes early diagnosis in humans. During 1980-1996, screening was done with serology and surgery was the unique choice of treatment. Since 1997, ultrasound (US) has been the method of choice for screening, and new choices of treatment for asymptomatic carriers are discussed in the CE guidelines. Methods Between 1997 and 2016, 42 734 abdominal USs were performed, 192 new asymptomatic cases were diagnosed and underwent a protocol according to the size, location and type of cyst. Treatment options included active surveillance (US monitoring, 83 [43.3%]), antiparasitic (albendazole, 92 [47.9%]) and surgery (17 [8.8%], including percutaneous treatment). Results After 7.7 y of follow-up, of the cases under active surveillance, 28 (33.7%) had to change treatment: 5 (6%) to surgery and 22 (26.5%) to albendazole. Of the patients treated with albendazole, 3 (3.2%) were operated on and 13 (14%) were treated with a second cycle of albendazole. Conclusion As a result of the present study, resolution of CE in a non-surgical way with albendazole is confirmed to be effective in asymptomatic carriers with CE1 or CE3a cysts. An update eliminates the strategy of active surveillance in type CE1 cysts

Background: Cystic echinococcosis (CE) is a parasitic zoonosis caused by infection with the larval stage of Echinococcus granulosus sensu lato This study investigated the prevalence and potential risk factors associated with human CE in the towns and rural areas of Ñorquinco and Ramos Mexia, Rio Negro province, Argentina. Methods: In order to detect abdominal CE cysts, we screened 892 volunteers by ultrasound and investigated potential risk factors for CE using a standardized questionnaire. Bivariate and multivariate analyses were used to estimate the Prevalence Ratio (PR) and their 95% CIs of the association between CE and the factors investigated. Results: Abdominal CE was detected in 42/892 screened volunteers (4.7%, CI 3.2-6.1), only two of who being under 15 years of age. Thirteen CE (30.9%) cases had 25 cysts in active stages (CE1, CE2, CE3) The most relevant risk factors identified in the bivariate analysis included: live in rural area (p=0.003), age >40 years (p=0.000), drinking always water of natural source (p=0.007), residing in rural areas during first five years of life (p=0.000) and live more than 20 years at your current address (p=0.013). In the multivariate model, statistically significant risk factors were: frequently touch dogs (p=0.012), residing in rural areas during first five years of life (p=0.004), smoking (p=0.000), age > 60 years (p­­=0.002) and live in rural areas (p=0.017).Conclusions: our results point toward infection with CE being acquired since childhood and with constant exposure throughout life, especially in rural areas with a general environmental contamination

Cystic echinococcosis is endemic in the Rio Negro province of Argentina and, for this, a control program using praziquantel in dogs was developed from 1980. The transmission rate to humans and sheep has decreased significantly, however transmission persists. In 2009 the vaccination of sheep with EG95 was incorporated in some areas of the province. The objective of the study was to evaluate the humoral responses to the vaccine EG95. Lambs received two vaccinations with the EG95 vaccine followed by a single booster injection when the animals were 1-1.5 years of age. Blood samples from 6 vaccinated groups and 4 no vaccinated sheep were obtained for determination of antibody titles against EG95 protein. Anti-EG95 responses were determined as described by Heath and Koolaard, 2012. Responses were evaluated from 331 animals. Median ELISA absorbance values in vaccinated group was 0.828, and in non-vaccinated groups was 0.218. EG95 antibody responses in sheep from different cohorts of non-immunized control groups and vaccinated groups reveal the sustained increase in response seen in animals following the third immunization. (p< 0.0001). Significant differences are also evidenced in ANOVA test (p< 0.001). An anti-EG95 antibody response was induced in all groups of immunized sheep, 4 times higher (0.828) than the mean observed in the control groups (0.218). Data described here indicate that following a third vaccination with the EG95 vaccine at 1 year of age the specific IgG responses detected in the serum of sheep increased to a level greater than that seen following the second immunization and that this response was maintained longitudinally over time, for at least 5 years. Fil: Larrieu, Edmundo Juan. Universidad Nacional de La Pampa. Facultad de Ciencias Veterinarias; Argentina. Universidad Nacional de Río Negro; Argentina Fil: Poggio, Thelma Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Ciencia y Tecnología "Dr. César Milstein". Fundación Pablo Cassará. Instituto de Ciencia y Tecnología ; Argentina Fil: Mujica, Guillermo. Provincia de Río Negro. Ministerio de Salud; Argentina Fil: Gauci, Charles G.. University of Melbourne; Australia Fil: Labanchi, Jose Luis. Provincia de Río Negro. Ministerio de Salud; Argentina Fil: Herrero, Eduardo. Provincia de Río Negro. Ministerio de Salud; Argentina Fil: Araya, Daniel. Provincia de Río Negro. Ministerio de Salud; Argentina Fil: Grizmado, Claudia. Provincia de Río Negro. Ministerio de Salud; Argentina Fil: Calabro, Arnoldo. Provincia de Río Negro. Ministerio de Salud; Argentina Fil: Talmon, Gabriel. Provincia de Río Negro. Ministerio de Salud; Argentina Fil: Crowley, Pablo Ernesto. Universidad Nacional de Río Negro; Argentina Fil: Santillán, Graciela. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud "Dr. C. G. Malbran". Departamento de Parasitología; Argentina Fil: Vizcaychipi, Katherina. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud "Dr. C. G. Malbran". Departamento de Parasitología; Argentina Fil: Seleiman, Marcos. Provincia de Río Negro. Ministerio de Salud; Argentina Fil: Sepulveda, Luis. Provincia de Río Negro. Ministerio de Salud; Argentina Fil: Arezo, Marcos. Provincia de Río Negro. Ministerio de Salud; Argentina Fil: García Cachau, Mariela Alejandra. Universidad Nacional de La Pampa. Facultad de Ciencias Veterinarias; Argentina Fil: Lamberti, Roberto. Universidad Nacional de La Pampa. Facultad de Ciencias Veterinarias; Argentina Fil: Molina, Leonardo. Universidad Nacional de La Pampa. Facultad de Ciencias Veterinarias; Argentina Fil: Gino, Lilia Mabel. Universidad Nacional de La Pampa. Facultad de Ciencias Veterinarias; Argentina Fil: Donadeu, Meritxell. University of Melbourne; Australia Fil: Lightowlers, Marshall W.. University of Melbourne; Australia

Cystic echinococcosis (CE) is a parasitic zoonosis caused by Echinococcus granulosus. The control program of CE of Rio Negro province, Argentina, involves annual surveillance using ultrasound (US) screening in school children, and five-year cross-sectional surveys to detect livestock farms with parasitized dogs by coproELISA with confirmation tests (Western Blot or PCR). Control program is based on deworming of dogs with praziquantel and the aim is to identify areas at risk of Cystic echinococcosis transmission to humans, using all available data sources. The information was spatially distributed in 13 program areas and, at a smaller geographical scale, in 80 Primary Health Care Centers. CoproELISA surveys involved three randomized sampling periods (2003-05, 2009-10, 2017-18), with 1790 canine fecal samples. The US surveys were conducted in 2003-08, 2009-16 and 2017-18 in 34,515 children. Heat maps were created at the smallest geographic scale with QGIS 3.4.6. For the consecutive sampling periods, prevalence of positive canine fecal samples from livestock farms were 14.7, 12.1 and 7.8%, respectively, and children prevalence was 0.4, 0.2 and 0.1%, respectively. The study has been developed on a scale according to which the temporal-spatial distribution of CE allows to adjust control strategies in those areas of potential transmission of the zoonosis to humans.

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.

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 vaccine's 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.

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., Author Summary Hydatid disease, otherwise known as cystic echinococcosis (CE), is caused by Echinococcus granulosus. The disease is common in many pastoral areas, including parts of the Rio Negro province of Argentina, and is formally recognised by the WHO as a Neglected Tropical Disease. We undertook the first scientific evaluation of the EG95 vaccine against transmission of hydatid disease in a field situation. Sheep in remote indigenous communities of Rio Negro were vaccinated over a six year period. Young lambs received two injections of vaccine and a single booster injection as one year old sheep. A similar region acted as a control where vaccination was not implemented. Evaluation of the outcomes of vaccination revealed a significant, 62% decrease in the prevalence of hydatid infection in 6 year old sheep. Our study provides a template for the application of vaccination in future efforts to control transmission of E. granulosus and reduce the burden of human disease caused by the parasite.