Genomic and eco-epidemiological investigations in Uruguay reveal local Chikungunya virus transmission dynamics during its expansion across the Americas in 2023.

Uruguay experienced its first Chikungunya virus outbreak in 2023, resulting in a significant burden to its healthcare system. We conducted analysis based on real-time genomic surveillance (30 novel whole genomes) to offer timely insights into recent local transmission dynamics and eco-epidemiological factors behind its emergence and spread in the country.

Genomics-based timely detection of Dengue Virus type I genotypes I and V in Uruguay.

We report the first whole-genome sequences of Dengue Virus type I genotypes I and V from Uruguay, including the first cases ever reported in the country. Through timely genomic analysis, identification of these genotypes was possible, aiding in timely public health responses and intervention strategies to mitigate the impact of dengue outbreaks.

Genomic and eco-epidemiological investigations in Uruguay reveal local Chikungunya virus transmission dynamics during its expansion across the Americas in 2023.

Uruguay experienced its first Chikungunya virus outbreak in 2023, resulting in a significant burden to its healthcare system. We conducted analysis based on real-time genomic surveillance (30 novel whole genomes) to offer timely insights into recent local transmission dynamics and eco-epidemiological factors behind its emergence and spread in the country.

A Multiplex Nanopore Sequencing Approach for the Detection of Multiple Arboviral Species.

The emergence and continued geographic expansion of arboviruses and the growing number of infected people have highlighted the need to develop and improve multiplex methods for rapid and specific detection of pathogens. Sequencing technologies are promising tools that can help in the laboratory diagnosis of conditions that share common symptoms, such as pathologies caused by emerging arboviruses. In this study, we integrated nanopore sequencing and the advantages of reverse transcription polymerase chain reaction (RT-PCR) to develop a multiplex RT-PCR protocol for the detection of Chikungunya virus (CHIKV) and several orthoflaviviruses (such as dengue (Orthoflavivirus dengue), Zika (Orthoflavivirus zikaense), yellow fever (Orthoflavivirus flavi), and West Nile (Orthoflavivirus nilense) viruses) in a single reaction, which provides data for sequence-based differentiation of arbovirus lineages.

Genome Sequences of SARS-CoV-2 P.1 (Variant of Concern) and P.2 (Variant of Interest) Identified in Uruguay.

Two severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants associated with increased transmission and immune evasion, P.1 and P.2, emerged in Brazil and spread throughout South America. Here, we report genomes corresponding to these variants that were recently detected in Uruguay. These P.1 and P.2 genomes share all substitutions that are characteristic of these variants.

First inter-laboratory comparison of Echinococcus granulosus sensu lato diagnosis in Latin America

[ABSTRACT]. Objective. To compare the performance of polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) tests for diagnosing Echinococcus granulosus in dog feces among national reference laboratories in Argentina, Chile, Peru, and Uruguay. Methods. National laboratories affiliated with the Ministry of Health/Agriculture of each country exchanged panels of 10 positive/negative samples obtained from their regular national surveillance programs in November 2015 – November 2016. All laboratories applied PCR; two also applied ELISA techniques. Sensitivity and specificity were determined for each laboratory and concordance of results among the laboratories was evaluated by Cohen Kappa coefficient. Results. Poor concordance (3 of 10 paired comparisons had values of Kappa > 0.4), low sensitivity and specificity across all laboratories, and poor performance of both techniques in detecting E. granulosus in canine feces was demonstrated in this study. An ex-post comparison of the laboratories’ test protocols showed substantial heterogeneity that could partially explain poor concordance of results. Conclusion. The results underscore the heterogeneity of canine echinococcosis diagnosis across the region and indicate possible sources of variability. Efforts to standardize canine echinococcosis testing must be included in the plan of action for the Regional Initiative for the Control of Cystic Echinococcosis. Future comparisons with fecal samples of known parasite load are needed.

[RESUMEN]. Objetivo. Comparar el rendimiento de los ensayos de la reacción en cadena de la polimerasa y el enzimoinmunoanálisis de adsorción en fase sólida (o ELISA, por su sigla en inglés) para diagnosticar Echinococcus granulosus en heces caninas en los laboratorios de referencia nacionales de Argentina, Chile, Perú y Uruguay. Métodos. Los laboratorios nacionales, afiliados a los ministerios de salud y agricultura y ganadería de cada país, intercambiaron paneles de diez muestras positivas y negativas obtenidas de sus respectivos programas nacionales de vigilancia desde el mes de noviembre del año 2015 hasta el mismo mes del año siguiente. Todos los laboratorios emplearon la reacción en cadena de la polimerasa y dos emplearon también técnicas de ensayo inmunoenzimático (ELISA). Se determinó la sensibilidad y la especificidad de cada laboratorio y se evaluó la concordancia entre los resultados de los laboratorios mediante el coeficiente kappa de Cohen. Resultados. Este estudio descubrió una escasa concordancia (3 de 10 comparaciones de pares obtuvieron valores de kappa > 0,4), una sensibilidad y especificidad bajas en todos los laboratorios y un rendimiento deficiente de ambas técnicas de diagnóstico de Echinococcus granulosus en heces caninas. La comparación ex post de los protocolos de ensayo de los laboratorios mostró una heterogeneidad sustancial que podría explicar parcialmente la escasa concordancia de los resultados. Conclusiones. Los resultados subrayan la heterogeneidad del diagnóstico de equinococosis canina en toda la región e indican posibles fuentes de esta variabilidad. Deben incluirse medidas para estandarizar la prueba de equinococosis canina en el plan de acción de la Iniciativa Sudamericana para el Control de la Equinococosis Quística. En el futuro serán necesarias comparaciones adicionales con muestras fecales con una carga de parásitos conocida.

[RESUMO]. Objetivo. Comparar o desempenho dos métodos de reação em cadeia da polimerase (PCR) e ensaio imunoenzimático (ELISA) no diagnóstico de infecção pelo Echinococcus granulosus em fezes de cães entre laboratórios de referência nacional na Argentina, Chile, Peru e Uruguai. Métodos. Laboratórios nacionais conveniados ao Ministério da Saúde/Agricultura de cada país participante intercambiaram grupos de 10 amostras positivas/negativas coletadas rotineiramente pelos programas nacionais de vigilância no período de novembro de 2015 a novembro de 2016. Todos os laboratórios empregaram o método de PCR e dois empregaram também o método de ELISA. A sensibilidade e a especificidade dos métodos foram determinadas em cada laboratório, e a concordância dos resultados entre os laboratórios participantes foi avaliada com o coeficiente kappa de Cohen. Resultados. Observou-se fraca concordância (3 de 10 comparações pareadas com kappa >0,4), baixa sensibilidade e especificidade e fraco desempenho de ambos os métodos na identificação do E. granulosus em amostras fecais de cães nos laboratórios participantes do estudo. Uma comparação retroativa revelou considerável heterogeneidade dos protocolos de análise laboratorial, o que poderia em parte explicar a fraca concordância entre os resultados. Conclusões. Os resultados deste estudo apontam para a falta de uniformidade no diagnóstico de equinococose canina em toda a Região e indicam possíveis causas para variabilidade. A padronização da análise laboratorial da equinococose canina deve constar do plano de ação para a Iniciativa Regional para Controle da Hidatidose. Outras comparações de amostras fecais de parasitas conhecidos devem ser realizadas.

First inter-laboratory comparison of Echinococcus granulosus sensu lato diagnosis in Latin America.

OBJECTIVE: To compare the performance of polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) tests for diagnosing Echinococcus granulosus in dog feces among national reference laboratories in Argentina, Chile, Peru, and Uruguay. METHODS: National laboratories affiliated with the Ministry of Health/Agriculture of each country exchanged panels of 10 positive/negative samples obtained from their regular national surveillance programs in November 2015 - November 2016. All laboratories applied PCR; two also applied ELISA techniques. Sensitivity and specificity were determined for each laboratory and concordance of results among the laboratories was evaluated by Cohen Kappa coefficient. RESULTS: Poor concordance (3 of 10 paired comparisons had values of Kappa > 0.4), low sensitivity and specificity across all laboratories, and poor performance of both techniques in detecting E. granulosus in canine feces was demonstrated in this study. An ex-post comparison of the laboratories' test protocols showed substantial heterogeneity that could partially explain poor concordance of results. CONCLUSION: The results underscore the heterogeneity of canine echinococcosis diagnosis across the region and indicate possible sources of variability. Efforts to standardize canine echinococcosis testing must be included in the plan of action for the Regional Initiative for the Control of Cystic Echinococcosis. Future comparisons with fecal samples of known parasite load are needed.

[Detection of stool antigens of Echinococcus granulosus in dogs belonging to slaughterhouse workers and offal merchants in Metropolitan Lima]. / Detección de coproantígenos de Echinococcus granulosus en canes de trabajadores de camales y comercializadores de vísceras en Lima metropolitana.

OBJECTIVE: To demonstrate the presence of Echinoccocus granulosus in the definitive host in the city of Lima, Perú, by detecting parasite antigens in the stool of dogs belonging to offal handlers and merchants in authorized slaughterhouses in Metropolitan Lima. METHODS: Stool samples were collected from 58 dogs and examined using the coproELISA technique for the detection of secretory/excretory antigens of E. granulosus. A survey was conducted to obtain information on pet feeding and handling practices. RESULTS: Positivity to E. granulosus was detected in 13.8% (8/58) of the dogs. In 27.8% (5/18) of the homes, at least one animal showed positivity, and in families that had more than four dogs the chances of finding positivity in at least one dog were higher (P < 0.05). In all homes where at least one dog tested positive the pets were fed on offal. Of study participants, 94.4% (17) knew nothing about the routes of transmission of hydatid disease. CONCLUSIONS: Results show the presence of definitive hosts in the urban area of Lima and underscore the need to more widely disseminate practices for the prevention of parasite transmission.

Detección de coproantígenos de Echinococcus granulosus en canes de trabajadores de camales y comercializadores de vísceras en Lima metropolitana / Detection of stool antigens of Echinococcus granulosus in dogs belonging to slaughterhouse workers and offal merchants in Metropolitan Lima

Objetivo. Demostrar la presencia de Echinoccocus granulosus en el hospedero definitivo en la ciudad de Lima, Perú, mediante la detección de antígenos del parásito en heces de canes pertenecientes a trabajadores y comercializadores de vísceras de centros de beneficio autorizados en Lima metropolitana. Métodos. Se recolectaron muestras de heces de 58 canes, que fueron evaluadas utilizando la técnica coproELISA para detectar antígenos secretorio/excretorio de E. granulosus. Mediante una encuesta se obtuvo información sobre las prácticas de alimentación y el manejo de las mascotas. Resultados. El 13,8% (8/58) de canes fue positivo a E. granulosus. En 27,8% (5/18) de los hogares se encontró al menos un animal positivo y se estimó que en las familias que tenían más de cuatro canes las posibilidades de encontrar al menos uno positivo eran mayores. En todos los hogares con al menos un can positivo sus mascotas se alimentaban con vísceras. El 94,4% (17) de los participantes no tenía conocimiento de las formas de contagio de la equinococosis. Conclusiones. Los resultados muestran la presencia de hospederos definitivos en la zona urbana de Lima y subrayan la necesidad de aumentar la difusión de las prácticas para evitar la transmisión del parasito.

Objective. To demonstrate the presence of Echinoccocus granulosus in the definitive host in the city of Lima, Perú, by detecting parasite antigens in the stool of dogs belonging to offal handlers and merchants in authorized slaughterhouses in Metropolitan Lima. Methods. Stool samples were collected from 58 dogs and examined using the coproELISA technique for the detection of secretory/excretory antigens of E. granulosus. A survey was conducted to obtain information on pet feeding and handling practices. Results. Positivity to E. granulosus was detected in 13.8% (8/58) of the dogs. In 27.8% (5/18) of the homes, at least one animal showed positivity, and in families that had more than four dogs the chances of finding positivity in at least one dog were higher (P < 0.05). In all homes where at least one dog tested positive the pets were fed on offal. Of study participants, 94.4% (17) knew nothing about the routes of transmission of hydatid disease. Conclusions. Results show the presence of definitive hosts in the urban area of Lima and underscore the need to more widely disseminate practices for the prevention of parasite transmission.

Detección de coproantígenos de Echinococcus granulosus en canes de trabajadores de camales y comercializadores de vísceras en Lima metropolitana / Detection of stool antigens of Echinococcus granulosus in dogs belonging to slaughterhouse workers and offal merchants in Metropolitan Lima

RESUMEN Objetivo Demostrar la presencia de Echinoccocus granulosus en el hospedero definitivo en la ciudad de Lima, Perú, mediante la detección de antígenos del parásito en heces de canes pertenecientes a trabajadores y comercializadores de vísceras de centros de beneficio autorizados en Lima metropolitana. Métodos Se recolectaron muestras de heces de 58 canes, que fueron evaluadas utilizando la técnica coproELISA para detectar antígenos secretorio/excretorio de E. granulosus. Mediante una encuesta se obtuvo información sobre las prácticas de alimentación y el manejo de las mascotas. Resultados El 13,8% (8/58) de canes fue positivo a E. granulosus. En 27,8% (5/18) de los hogares se encontró al menos un animal positivo y se estimó que en las familias que tenían más de cuatro canes las posibilidades de encontrar al menos uno positivo eran mayores. En todos los hogares con al menos un can positivo sus mascotas se alimentaban con vísceras. El 94,4% (17) de los participantes no tenía conocimiento de las formas de contagio de la equinococosis. Conclusiones Los resultados muestran la presencia de hospederos definitivos en la zona urbana de Lima y subrayan la necesidad de aumentar la difusión de las prácticas para evitar la transmisión del parasito.

ABSTRACT Objective To demonstrate the presence of Echinoccocus granulosus in the definitive host in the city of Lima, Perú, by detecting parasite antigens in the stool of dogs belonging to offal handlers and merchants in authorized slaughterhouses in Metropolitan Lima. Methods Stool samples were collected from 58 dogs and examined using the coproELISA technique for the detection of secretory/excretory antigens of E. granulosus. A survey was conducted to obtain information on pet feeding and handling practices. Results Positivity to E. granulosus was detected in 13.8% (8/58) of the dogs. In 27.8% (5/18) of the homes, at least one animal showed positivity, and in families that had more than four dogs the chances of finding positivity in at least one dog were higher (P < 0.05). In all homes where at least one dog tested positive the pets were fed on offal. Of study participants, 94.4% (17) knew nothing about the routes of transmission of hydatid disease. Conclusions Results show the presence of definitive hosts in the urban area of Lima and underscore the need to more widely disseminate practices for the prevention of parasite transmission.

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.

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 monoclonal antibody-based copro-ELISA kit for canine echinococcosis to support the PAHO effort for hydatid disease control in South America.

Cystic echinococcosis is still a major concern in South America. While some regions show advances in the control of the disease, others have among the highest incidence in the world. To reverse this situation the Pan American Health Organization (PAHO) has launched a regional project on cystic echinococcosis control and surveillance. An early concern of the program was the lack of a standardized diagnostic tool to monitor infection in dogs, a key target of control programs. Under this premise, we have developed a new copro-ELISA test after extensive screening of a large panel of monoclonal antibodies (MAbs) and polyclonal sera, which performs with high standards of sensitivity (92.6%) and specificity (86.4%) as established by necropsy diagnosis of dogs. The key component of the test, MAbEg9 has a convenient IgG isotype and reacts with a periodate-resistant epitope found in high molecular weight components of the worm. Time-course analysis of experimentally infected dogs showed that even animals with a very low number of parasites could be detected as early as day 20 post infection. The test was formulated in a ready-to-use kit format with proven stability of each component for a minimum of 3 months at room temperature. This characteristic facilitates its standardized use and shipping to other laboratories, which was demonstrated by the identical results obtained by two different laboratories in Peru and our own laboratory when a large number of field samples were analyzed independently in a blind fashion.