Analysis of vaccination strategy against cystic echinococcosis developed in the Province of Río Negro, Argentina: 12 years of work.

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. OBJECTIVE: To identify causes which can still generate infection in dogs, and to detect species/genotypes in circulation in the province of Río Negro. MATERIALS AND METHODS: 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. RESULTS: 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. CONCLUSION: 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.

Identification of potential 'hot spots' of cystic echinococcosis transmission in the province of Río Negro, Argentina.

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.

Epidemiology, diagnosis, treatment and follow-up of cystic echinococcosis in asymptomatic carriers.

Background: Río 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 <3 cm and is replaced by treatment with antiparasitic in all asymptomatic cases with CE1 or CE3a cysts <10 cm. The update also limits follow-up to 12-18 months to evaluate those cases with non-response to antiparasitic and switch to a surgical option.

Pilot field trial of the EG95 vaccine against ovine cystic echinococcosis in Rio Negro, Argentina: 8 years of work.

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.

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.

[Cystic echinococcosis in dogs and children in the province of Río 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.

Vigilancia de la equinococosis quística en perros y niños en la provincia de Río Negro, Argentina / Cystic echinococcosis in dogs and children in the province of Río Negro, Argentina

La equinococosis quística (EQ) es una enfermedad endémica en la provincia de Río Negro, Argentina. El programa de control de esta afección desarrolla sistemas de vigilancia epidemiológica basados en las técnicas de coproELISA/Western blot (WB) en muestras de heces de perros recolectadas del suelo, a fin de determinar establecimientos ganaderos (unidades epidemiológicas o UE) con transmisión presente. El objetivo de esta investigación fue evaluar la prevalencia de EQ en establecimientos ganaderos y su relación con la presencia de casos de EQ en niños de 0 a 14 años. Se seleccionaron aleatoriamente UE donde se obtuvieron muestras de materia fecal canina, las que fueron procesadas mediante coproELISA/WB. Asimismo, se identificaron casos nuevos ocurridos en niños del rango etario señalado. Se obtuvieron 571 muestras de 278 UE; 37 (6,5 %) fueron positivas a coproELISA/WB de 37 (13,3 %) UE con transmisión presente. Las diferencias con el relevamiento efectuado en el período 2003-2004 resultaron no significativas (p = 0,9), mientras que las diferencias con el relevamiento efectuado en UE de poblaciones originarias en 2009-2010 resultaron significativas (p = 0,02). Con relación a la densidad animal en el área de trabajo, las UE que dieron un resultado negativo tuvieron un promedio de 2 (DE: 2,1) perros por UE, en las UE con infección presente el promedio fue de 3 (DE 4,2), esta diferencia fue estadísticamente significativa (p = 0,02). Se diagnosticaron 12 casos en niños menores de 15 años. Se estableció que los casos de hidatidosis, en promedio, se situaron a menor distancia de los campos positivos a coproELISA/WB que de los campos negativos (p= 0,00307). La prueba de coproELISA/WB permitió identificar la dispersión de EQ en establecimientos ganaderos y analizar su relación con la ocurrencia de casos en niños.(AU)

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.(AU)

Vigilancia de la equinococosis quística en perros y niños en la provincia de Río Negro, Argentina / Cystic echinococcosis in dogs and children in the province of Río Negro, Argentina

La equinococosis quística (EQ) es una enfermedad endémica en la provincia de Río Negro, Argentina. El programa de control de esta afección desarrolla sistemas de vigilancia epidemiológica basados en las técnicas de coproELISA/Western blot (WB) en muestras de heces de perros recolectadas del suelo, a fin de determinar establecimientos ganaderos (unidades epidemiológicas o UE) con transmisión presente. El objetivo de esta investigación fue evaluar la prevalencia de EQ en establecimientos ganaderos y su relación con la presencia de casos de EQ en niños de 0 a 14 años. Se seleccionaron aleatoriamente UE donde se obtuvieron muestras de materia fecal canina, las que fueron procesadas mediante coproELISA/WB. Asimismo, se identificaron casos nuevos ocurridos en niños del rango etario señalado. Se obtuvieron 571 muestras de 278 UE; 37 (6,5 %) fueron positivas a coproELISA/WB de 37 (13,3 %) UE con transmisión presente. Las diferencias con el relevamiento efectuado en el período 2003-2004 resultaron no significativas (p = 0,9), mientras que las diferencias con el relevamiento efectuado en UE de poblaciones originarias en 2009-2010 resultaron significativas (p = 0,02). Con relación a la densidad animal en el área de trabajo, las UE que dieron un resultado negativo tuvieron un promedio de 2 (DE: 2,1) perros por UE, en las UE con infección presente el promedio fue de 3 (DE 4,2), esta diferencia fue estadísticamente significativa (p = 0,02). Se diagnosticaron 12 casos en niños menores de 15 años. Se estableció que los casos de hidatidosis, en promedio, se situaron a menor distancia de los campos positivos a coproELISA/WB que de los campos negativos (p= 0,00307). La prueba de coproELISA/WB permitió identificar la dispersión de EQ en establecimientos ganaderos y analizar su relación con la ocurrencia de casos en niños.

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.

Vigilancia de la equinococosis quística en perros y niños en la provincia de Río Negro, Argentina. / [Cystic echinococcosis in dogs and children in the province of 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.

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 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.