The Paraguayan Chaco is an isolated environment with its own unique ecosystem. In this region, Chagas disease remains a health problem. Chagas disease is caused by the parasite Trypanosoma cruzi, and it is primarily transmitted by triatomines. In order to identify the blood meal sources of triatomines, specimens of the vector were collected in domestic and peridomestic areas and the PCR-RFLP method was implemented. Cytochrome b was amplified from the samples and later subjected to digestion with two restriction enzymes: Hae III and Xho I.It was possible to generate distinct restriction patterns on the amplified material to identify several blood meal sources for the vectors. We employed the blood from several species as positive controls: human, chicken, canine, feline, and armadillo blood. However, we identified only 3 sources for the blood meals of the insect vectors: human, chicken and canine blood. In total, 76 triatomines were captured. T. cruzi was not found in any of them. In 61% of the captured specimens, the blood meal sources for the vectors could be identified. In 30% of these cases, the presence of DNA from more than one vertebrate was detected in the same triatomine. The most common blood meal source found was chicken blood. The presence of human and chicken blood in triatomines captured in domestic and peridomestic areas strongly suggests that the parasite can freely move amongst both areas regardless of food availability. Free vector movement in these areas constitutes an epidemiological threat for the inhabitants of the community under study.
Surra is a disease caused by the hemoflagellate pathogen Trypanosoma evansi which affects a wide variety of mammals. The only cases reported of the presence of T. evansi in Paraguay were identified in samples from horses and capybaras and they were detected by light microscopy in the 19th century. The aim of this study is to report three autochthonous cases of canine trypanosomiasis caused by T. evansi, for the first time in the country, using molecular techniques and their application for the differential diagnosis of trypanosomatids species. The technique implemented was real-time PCR-HRM, amplifying a fragment of the hsp70 gene, using a pair of primers initially used to discriminate Leishmania species. This is the first report on the usage of these primers to detect T. evansi through HRM analysis, which allows the differentiation of trypanosomatids species simultaneously, making it an efficient tool for differential diagnosis.
Background: Leishmaniasis is a vector-borne disease caused by a parasite protozoon from the genus Leishmania. Among the molecular techniques applied for detecting these parasites, real-time PCR with High Resolution Melting (PCR-HRM) proved advantageous since it simultaneously determines both the presence and species of the pathogen in one step, through amplification and later analysis of curves generated by melting temperature. Methods:Based on this molecular technique, the goal of this study was to estimate the PCR-HRM sensitivity for Leishmania spp. detection in different canine tissues by evaluating biological samples obtained from popliteal, submandibular, and pre-scapular lymph nodes, from bone marrow and ear pinnae of 28 stray dogs captured in the metropolitan area of Asunción (Paraguay). Results:The rk39 immunochromatographic test showed that 25/28 tested dogs (89%) presented antibodies against L. infantum. In 20/25 dogs that tested positive for rk39 (80%), it was possible to detect Leishmania spp. by PCR-HRM and determine that the species corresponded entirely to L. infantum. Regarding the analysis of different tissues, the parasite was detected in all popliteal lymph node samples, followed by high detection in submandibular (at 95%) and pre-scapular lymph nodes (at 90%), bone marrow (at 85%), and ear pinnae (at 85%). Conclusions:This study demonstrated that the use of real-time PCR-HRM using the molecular marker hsp70 was a highly sensitive method for simultaneously detecting and identifying Leishmania species in different tissues taken from infected dogs. In addition, the usefulness of ear pinnae as easily accessible tissue for molecular diagnosis was emphasized.
Leishmaniosis es un conjunto de enfermedades zoonóticas causadas por protozoos del género Leishmania sp. Desde tiempo atrás se vienen registrando numerosas infecciones en caninos y en los últimos años ocurrieron casos aislados en felinos. Algunos gatos que cohabitaron en áreas endémicas de leishmaniosis canina contrajeron la enfermedad. Es probable que el sistema inmunitario del felino sea capaz de controlar la contaminación del parásito, ya sea eliminándolo o bien manteniéndolo en un estado crónico subclínico. Se reporta un caso de leishmaniosis en un felino adulto joven, residente en Asunción (Paraguay), cuya sintomatología clínica era inespecífica. Los estudios de laboratorio permitieron identificar la presencia de amastigotes de Leishmania infantum. El paciente fue sometido a tratamiento con respuesta favorable y remisión total de signos clínicos. La identificación del protozooario es determinante para el diagnóstico de esta enfermedad. Si bien la leishmaniosis es menos frecuente en felinos que en caninos, debe incluirse en el diagnóstico diferencial, especialmente en áreas endémicas.
Cystic fibrosis (CF) is an autosomal recessive disorder and is caused by variants in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene. We aimed to study the frequency of the F508del variant, the most common variant worldwide, in patients with CF from Paraguay. The frequency of the F508del variant in Paraguayan patients with a clinical diagnosis of CF was assessed using a polymerase chain reaction followed by the sequencing of the PCR products. 43 of the 86 patients (50%) were homozygous for the F508del variant, 28 were heterozygous (32.56%), and the remaining 15 (17.44%) were non-carriers. In terms of alleles, there were 114 mutated (114/172 or 66.28%) and 58 did not correspond to this variant (58/172 or 33.72%). This is the first study of the frequency of the F508del variant in patients with CF in Paraguay. This information is of utmost relevance when planning and offering treatments from health services.
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