A novel loop-mediated isothermal amplification-based test for detecting Neospora caninum DNA

Background: Neospora caninum is a cyst-forming, coccidian parasite which is known to cause neurological disorders in dogs and abortion and neonatal mortality in cows and other livestock. This study reports the development of a loop-mediated isothermal amplification (LAMP) assay based on the Neospora caninum Nc-5 gene and compares its efficacy for detecting DNA to that of a semi-nested PCR test. Results: Six primers were designed based on the Nc-5 repeat region of N. caninum. Specific LAMP primers led to successful amplification of N. caninum DNA at 63 °C in 30 min. The LAMP assay was highly specific (i.e. it did not reveal cross-reactivity with other parasite species) and had a low N. caninum plasmid DNA limit of detection (1 fg), which is ten times higher than that for the semi-nested PCR. LAMP applicability was evaluated using a set of naturally-infected samples (59 from canine faeces and five from bovine abortions). Thirty-nine percent (25/64) of the naturally-infected samples were positive for N. caninum DNA by LAMP and 36% (23/64) by semi-nested PCR. However, the LAMP assay is much faster to perform than semi-nested PCR and provides results in 30 min. Conclusion: The optimized reaction conditions described in this study resulted in a sensitive, specific and rapid technique for detecting N. caninum DNA. Considering the advantages of LAMP for detecting N. caninum DNA, further assays aimed at testing its usefulness on a wider range of field samples are recommended. © 2017 The Author(s)

Criptosporidiosis y «Una Salud»

Criptosporidiosis es una enfermedad diarreica (zoonótica) causada por el parásito Cryptosporidium, el cual infecta al intestino. Se producen tanto en el ser humano como en diferentes animales de granja, mascotas y animales silvestres. Se subestima su papel y poco se conoce sobre su epidemiología, debido al escaso interés que suscita y la limitada disponibilidad de métodos de diagnóstico. La salud de las poblaciones tanto humana como animal, están relacionadas con una estrecha interacción de tres factores: el ambiente, los agentes patógenos, y las poblaciones, lo cual determina el estado de salud y enfermedad.El enfoque, conocido como «Una Salud», es una estrategia global en crecimiento que se está adoptando por una diversidad de organizaciones y responsables de las políticas en respuesta a la necesidad de enfoques integrados. Este enfoque puede ser relevante para una amplia gama de objetivos mundiales de desarrollo.Cada vez se habla más de salud integral, de salud holística. Parece que somos más conscientes de que la salud no se reduce a algo puramente biológico, sino que afecta a toda la persona. No es sólo ver al enfermo globalmente, es partir de la complejidad del ser humano y del mundo entero atravesado por la vulnerabilidad e interaccionar con la totalidad de los sujetos.Cryptosporidiosis is a diarrheal disease (zoonotic) caused by the parasite Cryptosporidium, which infects the intestine. It occurs in humans and in different pets, farm and wild animals. Its role is underestimated and little is known about its epidemiology, due to the low interest it draws and the limited availability of diagnostic methods. The population health, both human and animal, is related to a close interaction of three factors: the environment, pathogens, and population, which determines the state of health and disease; some of the diseases considered as emerging, that appeared in the last years.The approach, known as “One Health”, is a growing global strategy being adopted by a diversity of organizations and policy makers in response to the need for integrated approaches. This approach may be relevant to a wide range of global development goals.There is more and more talk about holistic health. It seems that we are more aware that health is not reduced to something purely biological, but affects the whole person. It is not only seeing the patient globally, it is starting from the complexity of the human being and the whole world crossed by vulnerability and interacting with the totality of the subjects

Neospora caninum: Relación Biológica con Toxoplasma gondii y su Potencial como Zoonosis

Objective. To carry out a systematic review of the literature on the zoonotic potential of Neospora caninum taking into account its biological relationship with Toxoplasma gondii. Materials and methods. A systematic review was carried out, using two databases for the collection of information (Science direct and Scielo), a search engine (Google Scholar) and the National University of Colombia Repository, all documents were organized through the bibliographic manager Mendeley for proper review. Key words like Neosporosis, Neospora caninum, “Toxoplasmosis”, “Toxoplasma gondii”, “Neospora humans”, “Neospora human infection”, “Neospora zoonotic” were used. A total of 8 articles were selected that look for the infection in humans. Results. Eight documents concerning the subject of Neospora in humans were used, whereas these reports show evidence of seropositivity in human samples of anti-N. caninum no studies were found that look for the isolation of the parasite in human tissues (by molecular or in vitro culture). Conclusions. According to the results of these studies, it is not ruled out that the existence of cross-reactions with T. gondii and therefore it should not be ruled out that Neosporosis can behave as a zoonosis. In this regard, more research is required to demonstrate exposure, associated clinical signs, or pathologies related to this infection.Objetivo. Realizar una revisión sistemática de la literatura sobre el potencial zoonótico de Neospora caninum teniendo en cuenta su relación biológica con Toxoplasma gondii. Materiales y métodos. Se realizó una revisión sistemática, utilizando dos bases de datos para la recopilación de la información (Science direct y Scielo), un buscador (Google Scholar) y el Repositorio de la Universidad Nacional de Colombia, todos los documentos fueron organizados por medio del gestor bibliográfico Mendeley para su correcta revisión. Se emplearon palabras claves como “Neosporosis”, “Neospora caninum”, “Toxoplasmosis”, “Toxoplasma gondii”, “Neospora humans”, “Neospora human infection”, “Neospora zoonotic”. Se encontraron ocho documentos que cumplían el criterio de evaluar la infección en humanos.  Resultado. Se utilizaron 8 documentos concernientes al tema de Neospora en humanos, donde se evidencia en algunos de estos reportes seropositividad en muestras de humanos, empleando diferentes técnicas serológicas para la detección de anticuerpos anti- N. caninum y anti-T. gondii, pero ninguna buscó la demostración directa del parásito en tejidos (por técnicas moleculares o cultivo). Conclusiones. Ningún estudio permite concluir definitivamente si existe infección en humanos, pues se utilizaron pruebas serológicas que tiene el riesgo de reacciones cruzadas con Toxoplasma gondii, se requieren estudios que demuestren exposición, signos clínicos asociados o patologías relacionadas con esta infección.

Aspectos económicos de la erradicación del gusano barrenador del ganado, cochliomyia hominivorax (coquere l, 1858), en colombia

La miasis causada por la mosca Cochliomyia hominivorax, no solo es desagradable y perjudicial, también puede facilitar procesos patológicos que conducen a la muerte del individuo afectado. Dentro de las estrategias de control y erradicación de esta enfermedad sobresale la técnica de insecto estéril, diseñada para afectar la eficiencia reproductiva de este parásito mediante la inducción de mutaciones letales dominantes en el ADN de las células sexuales de los machos que generan progenie no viable. El programa de erradicación de esta mosca ha sido un éxito en algunos países de América, pero a un costo elevado. La mayoría de los países, con la infestación natural de esta plaga, corresponden a los denominados en vías de desarrollo. En esta revisión se presentan los estudios efectuados sobre la evaluación económica de los programas de lucha, señalando a su vez la carencia de estudios detallados y continuos sobre la vigilancia epidemiológica y la evaluación económica de la erradicación del parásito, y de su biología, ecología y control en los países endémicos, lo cual en la práctica dificulta un análisis pormenorizado. La sostenibilidad del programa de eliminación demanda la obtención de recursos económicos suficientes por cuenta de capitales públicos, privados y extranjeros, que aseguren la viabilidad del mismo mediante la cooperación internacional, la participación activa de los productores, y una estructura sanitaria que garantice una vigilancia activa. Se deben analizar también el panorama político y otros aspectos de seguridad en el área rural, así como los aspectos ambientales. Una de las alternativas de control integrado debe incluir la adecuada vigilancia epidemiológica, tanto de los casos animales como de los seres humanos, el tratamiento individual oportuno con insecticidas, la implementación permanente del sistema SWASS y, en lo posible, la restricción de aquellas prácticas de manejo animal que ocasionan lesiones dérmicas.The screwworm myiasis (Cochliomyia hominivorax) is unpleasant and harmful zoonotic disease, and it could lead to death in appropriate conditions. From many control strategies, sterile insect technique stand out because its design affects parasite reproductive efficiency through induction of dominant lethal mutations on DNA inside male germ cells producing non viable offspring. Screwworm eradication program has been successful in some countries of Americas, but its costs are considerable. Mostly of countries naturally infested are named in development. In this review, studies of economical evaluation about area-wide programs control are presented, pointed out lack of detailed and continual information as epidemiologic surveillance and economical evaluation of screwworm eradication as its biology, ecology and control in endemic countries that prevents detailed analysis. Eradication program sustainable could demands to get enough economical support of public, particular and foreign capitals. They will secure its viability through international cooperation, active participation of animal producers and sanitary structure for warranty active surveillance. Political panorama and other aspects of rural security should be analyzed. Environmental aspects must be analyzed. An integrated control alternative of screwworm should include epidemiological surveillance appropriate of animal and human cases, timely insecticide treatment of individual cases, permanent carrying out of SWASS method and, as possible as it can be, restriction of injure livestock practices causing dermical lesions

Distribución de garrapatas Rhipicephalus (Boophilus) microplus en bovinos y fincas del Altiplano cundiboyacense (Colombia)

In Colombia, the presence of Rhipicephalus (Boophilus) microplus ticks has been registered in altitudes ranging from zero to 2,600 meters above the sea level. Currently, there are no reports of its distribution in the Altiplano Cundiboyacense, an important dairy region of Colombia. In the present work, ticks of each stage were searched on cattle and pastures from 108 farms ranging from 2,000 to 3,000 meters of altitude. A total of 851 specimens of R. (B.) microplus (100% of total) were collected from 34 farms located between 1,966 and 2,903 meters of altitude. This is the first report in Colombia on the presence of this tick in altitudes higher than 2,600 meters and on its distribution in the Altiplano Cundiboyacense. These results confirm the existence of this tick as an ectoparasite of cattle, vector of haemoparasites, in areas where it was no previously reported, and they present a potential risk of transmission of these pathogens in highly susceptible bovine populations. Future research must define the bioecology and population dynamics of this tick in that region and the prevalence of cattle infected with tick-borne pathogens.En Colombia, la presencia de la garrapata Rhipicephalus (Boophilus) microplus ha sido registrada en altitudes que oscilan entre 0 y 2.600 metros sobre el nivel del mar (msnm). Actualmente, no existen reportes de distribución de esta especie en el Altiplano cundiboyacense, zona geográfica de importancia nacional en la producción de leche. Para el presente trabajo se buscaron especímenes de cada uno de los estadios de la garrapata en bovinos y en potreros de 108 predios situados entre los 2.000 y 3.000 msnm, en las diferentes zonas del Altiplano cundiboyacense. Se colectaron 851 garrapatas, en total, de la especie R. (B.) microplus (100% de las muestras) en 34 predios ubicados entre los 1.966 y 2.903 msnm Este es el primer estudio en Colombia que registra la presencia de esta garrapata a una altitud superior a los 2.600 msnm y que caracteriza su distribución en el Altiplano cundiboyacense. Los resultados obtenidos demuestran la existencia de esta especie, ectoparásito de bovinos y vector de hemoparásitos, en zonas geográficas donde hasta hace algunos años no se registraba su presencia. Lo anterior plantea un panorama de riesgo para la presentación de babesiosis y anaplasmosis bovina en poblaciones altamente susceptibles. Estudios posteriores deberán definir la bioecología de esta garrapata en esta región y la prevalencia de infección en bovinos de los patógenos asociados. 

Trypanosoma evansi in horses from the Colombia - Associated Infection Factors

Background: Trypanosoma evansi is the most common protozoan in tropical and subtropical regions of the world, due to its ability to maintain and be transmitted by vectors such as Stomoxys spp. and Tabanus spp. This protozoan causes high morbidity and mortality rates in horses in African, American and Asian countries. In the years 2021 and 2022, a high mortality rate was reported among horses with symptoms associated with Trypanosoma spp. in the municipality of Arauca, department of Arauca, Colombia. The investigation described here was therefore carried out, seeking to identifythe pathogens and risk factors that led to the death of the horses in this region of Colombia.Cases: Blood samples were collected from Colombian criollo horses and dogs, as were samples of ticks, flies and horseflies that infested the horses. A variety of tissue samples were removed from the horses a few min after their death for histopathological analysis. Two questionnaires were applied to obtain information about the horses and the environment in which they live. The results of the clinical examination revealed pale mucous membranes, jaundice, high fever, dehydration and lethargy. The horses were also infested with Amblyomma mixtum (17.6%) and Dermacentor nitens (82.4%) ticks, and with Tabanus pungens (74%), Tabanus spp. (26%), and Stomoxys calcitrans flies (100%), while the dogs were infested with Rhipicephalus sanguineus s.l. (77.7%) and Amblyomma mixtum (22.2%) ticks. The blood smear test results revealed the presence of Trypanosoma spp. in 66.6% (n = 4) of the horse blood samples, and in 50% (n = 1) of the dog blood samples. PCR performed to identify the Trypanosoma species confirmed the presence of T. evansi. Histological examination of the spleen revealed the involvement and dissemination of T. evansi in the tissues. The horses also showed the presence of Equine Infectious Anemia Virus (EIAV).Discussion: This is the first updated specific report of T. evansi in criollo horses in the savannah flood zone of the municipality of Arauca, Colombia. The main risk associated with T. evansi infection in horses was found to be infestation with the natural vector T. pungens and the mechanical vector S. calcitrans, which are efficient ectoparasites for the transmission of this parasite. The presence of T. evansi in dogs represents a constant risk to horses, because dogs may serve as a reservoir for the maintenance of the hemoparasite in the population under study. Another risk factor for horses could be the presence of vampire bats (Desmodus rotundus), a species of bat that has been described as a vector and reservoir of T. evansi in Colombia. The presence of EIAV antibodies in the horses under study can be attributed to the exposure of sick horses to vectors of this virus, such as Tabanus spp., S. calcitrans and inanimate needle-shaped fomites. This is the first study that identifies the coinfection of T. evansi and EIAV in horses in the floodplain region of Colombia. In view of the importance of these 2 pathologies to the health of horses, a greater number of tests and a larger animal population will be required to determine if this coinfection is the cause of the death of criollo horses in this region of Colombia. Lastly, the owners reported that pharmacological control with trypanocides has not been successful in most of the outbreaks that occurred during the years 2021 and 2022. This may suggest that Trypanosoma evansi is developing resistance to these drugs; therefore, specific studies will be required in the future to test this hypothesis.Keywords: Arauca, Equine Infectious Anemia Virus, floodplain savannah, mortality, surra

Prevalencia y factores de riesgo de cryptosporidium spp. y giardia spp. en terneros de ganado lechero de la zona noroccidental de la sabana de bogotá

Objetivo Estimar la prevalencia de Cryptosporidium spp. y Giardia spp. en terneros de 0 a 2 meses de ganado lechero de la zona noroccidental de la Sabana de Bogotá. Se estimaron los factores de riesgo de la producción lechera que puedan incurrir en la infección de personas y animales. Métodos Estudio de corte transversal, mediante la toma de materia fecal de terneros. Se determinó la prevalencia de punto para Giardia spp. y Cryptosporidium spp. a través los resultados de los coprológicos (Ritchie para Giardia spp. y Ziehl-Neelsen modificada para Cryptosporidium spp.). Se calcularon Odds Ratio para establecer los factores de riesgo asociados entre estos dos géneros de protozoarios y el manejo de las Buenas Prácticas Ganaderas. Resultados Se evaluaron 33 fincas dedicadas a la producción lechera, donde se muestrearon 308 terneros. La prevalencia para Giardia spp. fue de 37,3 %, 115 animales positivos y para Cryptosporidium spp. fue de 4,9 % 15, animales positivos. Conclusión Existe un foco de Giardia spp. y Cryptosporidium spp. en el noroccidente de la Sabana de Bogotá, del cual no se tenía conocimiento previo. La prevalencia de Giardia spp. para la región estudiada, está en el límite alto del rango reportado para Sudamérica. En el caso de Cryptosporidium spp. la situación es distinta, la prevalencia se encuentra en el limite bajo del rango reportado para Sudamérica.  Los factores de riesgo asociados a Giardia spp. y Cryptosporidium spp. en las explotaciones lecheras del noroccidente de la Sabana de Bogotá, dependen de Buenas Prácticas Ganaderas

Associated risk factors to myiasis by cochliomyia hominivorax on cattle farms in puerto Boyacá (Colombia)

Se realizó un estudio descriptivo-transversal en el municipio de Puerto Boyacá para identificar los factores de riesgo asociados a la miiasis por Cochliomyia hominivorax en bovinos. En 44 fincas ganaderas seleccionadas en un muestreo por conveniencia, se efectuó una entrevista personal a cada ganadero acerca del uso del suelo, sistema de producción ganadera y productos para el tratamiento de la miiasis traumática, entre el 6 y el 15 de febrero del 2007. El material suministrado para recolección de larvas fue recogido entre la última semana de abril y la primera semana de mayo del 2007. Análisis de frecuencias, medidas de asociación y pruebas no paramétricas fueron utilizados. Se detectaron 49 casos (0,4%) de miiasis por C. hominivorax en 12.325 bovinos registrados, en 18 (41%) de 44 fincas ganaderas. La causa de herida más frecuente fue abrasión por alambre de púas en 16 casos (32,7%). Los lugares de infestación más frecuente fueron el ombligo y el escroto con 9 casos (18,4%) cada uno. Se encontró asociación entre grupos etarios en bovinos y la presentación de casos por C. hominivorax (P=0,000; =0,05). Se halló asociación entre el sistema de producción bovina y la presentación de casos por C. hominivorax (P=0,044; =0,05). Se registró el uso de productos no adecuados en el tratamiento de la miiasis traumática como creolina (31 fincas), gasolina (4 fincas), aceite quemado (2 fincas), Baygon® (1 finca) y Vanodine® (1 finca). Se concluye que las fincas ganaderas seleccionadas compartieron factores de riesgo, según el sistema de producción ganadera. Se requiere intervención para evitar perjuicios en animales y consumidores por la utilización de productos no [email protected] Cross-sectional and descriptive study was carried out in countryside of Puerto Boyacá town for identification of Cochliomyia hominivorax myiasis risk factors on cattle. Fourty four cattle farms were selected with a not-random sampling. Personal interviews were carried out to livestock farmers. Land use, livestock production system, and products for treatment of screwworm myiasis were investigated on February 6-15 of 2007. Stuff and instructions were supplied for taking larvae from animal wounds afterwards register survey application. Those materials were collect since last week on April of 2007 until first week on May of 2007. Frequency analysis, cross-tabs measures and non parametric tests were used. 49 screwworm myiasis cases (0.4%) out of 12,325 registered cattle were found, in 18 (41%) out of 44 selected cattle farms. The main cause of wounds recorded was injury by barbed wire with 16 cases (32.7%). Body sites more frequently infested were navel and scrotum with 9 cases (18.4%) each one. Association between age group in cattle and screwworm myiasis cases was founded (P=0.000; =0.05). Association between cattle production system and screwworm myiasis cases was founded (P=0.044; =0.05). Usual handling of non appropriate products for screw worm myiasis treatment like “creolina” (31 farms), gasoline (4 farms), used motor lubricant oil (2 farms), Baygon® (1 farm) and Vanodine® (1 farm) was registered. It may be concluded that selected cattle farms shared risk factors, depending on cattle production system. It demands epidemiological intervention to avoid damages to animals and consumers for using of inappropriate products for treatment of traumatic myiasis

Distribución de garrapatas Rhipicephalus (Boophilus) microplus en bovinos y fincas del Altiplano cundiboyacense (Colombia)

En Colombia, la presencia de la garrapata Rhipicephalus (Boophilus) microplus ha sido registrada en altitudes que oscilan entre 0 y 2.600 metros sobre el nivel del mar (msnm). Actualmente, no existen reportes de distribución de esta especie en el Altiplano cundiboyacense, zona geográfica de importancia nacional en la producción de leche. Para el presente trabajo se buscaron especímenes de cada uno de los estadios de la garrapata en bovinos y en potreros de 108 predios situados entre los 2.000 y 3.000 msnm, en las diferentes zonas del Altiplano cundiboyacense. Se colectaron 851 garrapatas, en total, de la especie R. (B.) microplus (100% de las muestras) en 34 predios ubicados entre los 1.966 y 2.903 msnm Este es el primer estudio en Colombia que registra la presencia de esta garrapata a una altitud superior a los 2.600 msnm y que caracteriza su distribución en el Altiplano cundiboyacense. Los resultados obtenidos demuestran la existencia de esta especie, ectoparásito de bovinos y vector de hemoparásitos, en zonas geográficas donde hasta hace algunos años no se registraba su presencia. Lo anterior plantea un panorama de riesgo para la presentación de babesiosis y anaplasmosis bovina en poblaciones altamente susceptibles. Estudios posteriores deberán definir la bioecología de esta garrapata en esta región y la prevalencia de infección en bovinos de los patógenos asociados.  In Colombia, the presence of Rhipicephalus (Boophilus) microplus ticks has been registered in altitudes ranging from zero to 2,600 meters above the sea level. Currently, there are no reports of its distribution in the Altiplano Cundiboyacense, an important dairy region of Colombia. In the present work, ticks of each stage were searched on cattle and pastures from 108 farms ranging from 2,000 to 3,000 meters of altitude. A total of 851 specimens of R. (B.) microplus (100% of total) were collected from 34 farms located between 1,966 and 2,903 meters of altitude. This is the first report in Colombia on the presence of this tick in altitudes higher than 2,600 meters and on its distribution in the Altiplano Cundiboyacense. These results confirm the existence of this tick as an ectoparasite of cattle, vector of haemoparasites, in areas where it was no previously reported, and they present a potential risk of transmission of these pathogens in highly susceptible bovine populations. Future research must define the bioecology and population dynamics of this tick in that region and the prevalence of cattle infected with tick-borne pathogens

A novel loop-mediated isothermal amplification-based test for detecting Neospora caninum DNA

"Background: Neospora caninum is a cyst-forming, coccidian parasite which is known to cause neurological disorders in dogs and abortion and neonatal mortality in cows and other livestock. This study reports the development of a loop-mediated isothermal amplification (LAMP) assay based on the Neospora caninum Nc-5 gene and compares its efficacy for detecting DNA to that of a semi-nested PCR test. Results: Six primers were designed based on the Nc-5 repeat region of N. caninum. Specific LAMP primers led to successful amplification of N. caninum DNA at 63 °C in 30 min. The LAMP assay was highly specific (i.e. it did not reveal cross-reactivity with other parasite species) and had a low N. caninum plasmid DNA limit of detection (1 fg), which is ten times higher than that for the semi-nested PCR. LAMP applicability was evaluated using a set of naturally-infected samples (59 from canine faeces and five from bovine abortions). Thirty-nine percent (25/64) of the naturally-infected samples were positive for N. caninum DNA by LAMP and 36% (23/64) by semi-nested PCR. However, the LAMP assay is much faster to perform than semi-nested PCR and provides results in 30 min. Conclusion: The optimized reaction conditions described in this study resulted in a sensitive, specific and rapid technique for detecting N. caninum DNA. Considering the advantages of LAMP for detecting N. caninum DNA, further assays aimed at testing its usefulness on a wider range of field samples are recommended. © 2017 The Author(s).