Detection of Leishmania infantum kinetoplast DNA by Real Time PCR in hair of wild rabbits

The study of potential wild mammal reservoirs is necessary for the surveillance of leishmaniosis, as Leishmania protozoans have been isolated from a wide range of wild and domestic animal species, including Leporidae. Recently, it has been demonstrated that both hares and wild rabbits can act as sylvatic reservoirs of Leishmania. In Spain, most of the research involving wild rabbits has been developed in the central area of Madrid and in the southeastern Mediterranean coast. We studied the presence of Leishmania infantum in 116 wild rabbits (Oryctolagus cuniculus) captured in Santovenia de Pisuerga, Valladolid, Spain. Hair samples were analyzed by real time PCR. L. infantum kinetoplast DNA (kDNA) was detected and quantified in 4 out of 116 analyzed animals. The estimated number of parasites obtained were quite variable, ranging from 2.60 to 276.60. Hair samples can be collected by non-invasive methods, being a proper sample for Leishmania detection in wild Leporidae, which have an important role as reservoirs of Leishmania. Our findings enhance the need for more extensive studies in different geographical areas.S

Acuerdo diagnóstico entre equipos de emergencias y servicios hospitalarios Diagnostic agreement between emergency teams and hospital services

Objetivos: Desde hace 10 años, la Empresa Pública de Emergencias Sanitarias de Andalucía estudia la concordancia de sus equipos con la atención primaria y los hospitales. Se investiga el acuerdo diagnóstico entre estos equipos y los hospitalarios, y en caso de encontrar diferencias, se valora si éstas podrían estar en función del equipo de emergencias, el recurso de traslado o el centro hospitalario. Pacientes y método: Estudio descriptivo. Se incluyó a 510 pacientes (seleccionados al azar) cuyos datos se conocía, atendidos por equipos 061 de Málaga en 2001 y que precisaron traslado a algún hospital público. Se recabaron datos sobre la filiación, la asistencia recibida, el traslado, el centro hospitalario y el diagnóstico. El máximo de diagnósticos permitidos fue 3, codificados según la clasificación CIE-9 MC. Para la comparación se utilizó el índice kappa. Resultados: Se perdieron 10 casos. La media de diagnósticos del 061 por paciente fue de 1,48, mientras que en los informes hospitalarios fue de 1,59. Los más frecuentes hacían referencia a traumatismos y enfermedades cardiovasculares (los inespecíficos se aproximaron al 20%). El 59% de los pacientes tenía al menos un diagnóstico coincidente. Para un nivel de confianza del 95% obtuvimos un índice kappa de 0,478 (proporción de conformidad del 73,9%). Conclusiones: Obtuvimos una concordancia global moderada, con mejores resultados para el Equipo de Coordinación Avanzada y traslados en ambulancia convencional por la simplicidad de los diagnósticos. Sólo llegó a niveles considerados como «buenos» el Hospital Costa del Sol, con el que se comparten las guías de trabajo. El porcentaje de diagnósticos poco precisos fue alto. Las propuestas de mejora deberán ir desde la revisión de los modos de trabajo hasta la aplicación de nuevas tecnologías.Objectives: Over the last 10 years the Public Health Emergency Service of Andalusia (Spain) has been conducting a study into the diagnostic agreement among its teams (061 teams) and those of primary care and hospitals. Diagnostic agreement between these teams and hospital teams was evaluated. When discrepancies were found, an assessment was made of whether these corresponded to the emergency team, transfer resources or hospital. Patients and method: A descriptive study was performed. Five hundred ten patients whose particulars were already known were randomly selected. The patients, who required transfer to a public hospital, received assistance from 061 teams in Malaga in 2001. Data were gathered on personal details, the assistance received, transfer, hospital and diagnosis or diagnoses. The maximum number of diagnoses permitted was three, coded in accordance with the CIE-9 CM classification. The Kappa index was used for comparisons. Results: Ten cases were lost due to missing data. The mean number of diagnoses per patient was 1.48 for 061 teams and was 1.50 in hospital reports. The most common of diagnoses related to injuries and cardiovascular diseases (non-specific diagnoses accounted for approximately 20%). Fifty-nine percent of the patients had at least one diagnosis that coincided. We obtained κ = 0.478 for a confidence level of 95% (the agreement rate was 73.9%). Conclusions: Overall agreement was moderate, with better results in the Advanced Coordination Team and conventional ambulance transfer due to the simplicity of the diagnoses.Results classified as «good» were achieved only in the Hospital Costa del Sol, which uses working guidelines similar to those of the Public Health Emergency Service. The percentage of inexact diagnoses was high. Proposals for improvement should range from revising the working methods used to applying new technologies

New insights into experimental visceral leishmaniasis: Real-time in vivo imaging of Leishmania donovani virulence.

Visceral leishmaniasis is an insidious neglected disease with worldwide distribution. It is caused by parasites from the Leishmania donovani complex, which are able to be transmitted by different species of phlebotomine sand flies and to infect numerous mammal hosts. Despite the high number of people infected or at risk, and the remarkable quantity of studies focusing on this disease, a proper experimental model to efficiently decipher the infectious process of visceral leishmaniasis taking into account the nuances of parasite’s virulence and the duration of the infection is still lacking. Therefore, using golden Syrian hamsters and BALB/c mice, state-of-the-art genetic manipulation applied on a fully virulent L. donovani strain and in vivo imaging approaches, we describe herein three benefits for experimental visceral leishmaniasis: (i) the development of a double transfected bioluminescent (firefly luciferase) and fluorescent (E2-crimson) virulent strain of L. donovani (Ld1S_luci_E2-crimson), favoring a wide range of both in vivo and in vitro investigations, (ii) the establishment of a non-invasive mouse model to evaluate the infectious process during visceral leishmaniasis and the parasite’s virulence in real time, allowing longitudinal studies with the same animals, and (iii) the elaboration of a suitable method to reinstate (and verify anew) the virulence in a population of attenuated parasites, by recovering persistent parasites from chronic infected mice. Consequently, these results open up new perspectives on the study of visceral leishmaniasis, especially in the fields of therapeutics and vaccinology, since the model described herein renders now possible long-lasting follow up studies, with easy and accurate day-by-day verifications of the infection status along with a reduced number of laboratory animals.ClinicalTrials.gov 2013-0047