Systematic review on the biology, ecology, genetic diversity and parasite transmission potential of Panstrongylus geniculatus (Latreille 1811) in Latin America.

Panstrongylus geniculatus (Latreille, 1811) is the triatomine with the largest geographic distribution in Latin America. It has been reported in 18 countries from southern Mexico to northern Argentina, including the Caribbean islands. Although most reports indicate that P. geniculatus has wild habitats, this species has intrusive habits regarding human dwellings mainly located in intermediate deforested areas. It is attracted by artificial light from urban and rural buildings, raising the risk of transmission of Trypanosoma cruzi. Despite the wide body of published information on P. geniculatus, many knowledge gaps exist about its biology and epidemiological potential. For this reason, we analysed the literature for P. geniculatus in Scopus, PubMed, Scielo, Google Scholar and the BibTriv3.0 databases to update existing knowledge and provide better information on its geographic distribution, life cycle, genetic diversity, evidence of intrusion and domiciliation, vector-related circulating discrete taxonomic units, possible role in oral T. cruzi transmission, and the effect of climate change on its biology and epidemiology.

A phased genome assembly of a Colombian Trypanosoma cruzi TcI strain and the evolution of gene families.

Chagas is an endemic disease in tropical regions of Latin America, caused by the parasite Trypanosoma cruzi. High intraspecies variability and genome complexity have been challenges to assemble high quality genomes needed for studies in evolution, population genomics, diagnosis and drug development. Here we present a chromosome-level phased assembly of a TcI T. cruzi strain (Dm25). While 29 chromosomes show a large collinearity with the assembly of the Brazil A4 strain, three chromosomes show both large heterozygosity and large divergence, compared to previous assemblies of TcI T. cruzi strains. Nucleotide and protein evolution statistics indicate that T. cruzi Marinkellei separated before the diversification of T. cruzi in the known DTUs. Interchromosomal paralogs of dispersed gene families and histones appeared before but at the same time have a more strict purifying selection, compared to other repeat families. Previously unreported large tandem arrays of protein kinases and histones were identified in this assembly. Over one million variants obtained from Illumina reads aligned to the primary assembly clearly separate the main DTUs. We expect that this new assembly will be a valuable resource for further studies on evolution and functional genomics of Trypanosomatids.

The identification of two Trypanosoma cruzi I genotypes from domestic and sylvatic transmission cycles in Colombia based on a single polymerase chain reaction amplification of the spliced-leader intergenic region.

A single polymerase chain reaction (PCR) reaction targeting the spliced-leader intergenic region of Trypanosoma cruzi I was standardised by amplifying a 231 bp fragment in domestic (TcIDOM) strains or clones and 450 and 550 bp fragments in sylvatic strains or clones. This reaction was validated using 44 blind coded samples and 184 non-coded T. cruzi I clones isolated from sylvatic triatomines and the correspondence between the amplified fragments and their domestic or sylvatic origin was determined. Six of the nine strains isolated from acute cases suspected of oral infection had the sylvatic T. cruzi I profile. These results confirmed that the sylvatic T. cruzi I genotype is linked to cases of oral Chagas disease in Colombia. We therefore propose the use of this novel PCR reaction in strains or clones previously characterised as T. cruzi I to distinguish TcIDOMfrom sylvatic genotypes in studies of transmission dynamics, including the verification of population selection within hosts or detection of the frequency of mixed infections by both T. cruzi I genotypes in Colombia.

Aedes albopictus distribution in Ibagué, Colombia: Potential risk of arboviral outbreaks / Distribución de Aedes albopictus en Ibagué: potencial riesgo de brotes de arbovirosis

Introduction: Aedes albopictus is a vector for arboviruses, such as dengue, Zika, chikungunya, and yellow fever. The first A. albopictus reports on the American continent date back to 1985. It has spread rapidly throughout Colombia since its first report in 1998 due to its ecological and physiological adaptation capability. Objective: To determine A. albopictus distribution in the 13 communes of Ibagué, Colombia. Materials and methods: Samples were collected between May and November 2022 in the 13 communes of Ibagué. Vacuum sampling and sweep-netting entomological nets were used in areas with abundant vegetation. The mosquitoes were transported to the Laboratorio de Investigaciones en Parasitología Tropical at the Universidad del Tolima for taxonomic determination. Results: We identified 708 A. albopictus specimens distributed throughout Ibague's 13 communes. The highest vector abundance occurred in communes 10, 11, 7, 8, 2, and 9; communes 3, 4, 5, 6, 12, and 13 had a relative abundance of around 3%, while commune 1 had 2% of relative abundance. Conclusions: Aedes albopictus is distributed throughout all the communes of Ibague. Its dispersion has probably been favored by this region's environmental and social conditions. We recommend annual monitoring of these vectors populations and molecular characterization of the found arboviruses. Ascertaining this mosquito's distribution throughout the city will enable focusing entomological control strategies and preventing future arbovirus outbreaks.

Introducción: Aedes albopictus es un vector de arbovirus como dengue, Zika, chikungunya y fiebre amarilla. Los primeros reportes en el continente americano datan de 1985 y dada su capacidad de adaptación ecológica y fisiológica, se ha distribuido rápidamente en el territorio colombiano desde su primer reporte en 1998. OBJETIVO: Determinar la distribución de A. albopictus en las comunas de Ibagué, Colombia. Materiales y métodos: Los muestreos se realizaron entre  mayo y noviembre de 2022 en zonas con abundante  vegetación de las 13 comunas de Ibagué. Se emplearon aspiradores y redes entomológicas. Los mosquitos fueron transportados al Laboratorio de Investigaciones en  Parasitología Tropical de la Universidad del Tolima para su determinación taxonómica. RESULTADOS: Se identificaron 708 ejemplares de A.  lbopictus, distribuidos en las comunas de Ibagué. La mayor abundancia del vector se presentó en las comunas 10, 11, 7, 8, 2 y 9. Las comunas 3, 4, 5, 6, 12 y 13 presentaron abundancias relativas cercanas al 3 %, y la comuna 1 tuvo una abundancia del 2 %. CONCLUSIONES: Aedes albopictus está distribuido en todas las comunas de Ibagué, probablemente su dispersión se ha visto favorecida por las condiciones ambientales y sociales de esta región. Se recomienda hacer seguimiento anual a las poblaciones de este vector y realizar una caracterización molecular de los arbovirus encontrados. Además, el conocer la distribución de este mosquito en la ciudad permitirá focalizar las estrategias de control entomológico y prevenir futuros brotes de arbovirosis.

Molecular detection of Toxoplasma gondii in meat for human consumption in Ibague, Colombia / Detección molecular de Toxoplasma gondii en carnes para consumo humano en Ibagué, Colombia

Introduction: Toxoplasma gondii is a parasite with great zoonotic potential. It can infect a broad range of warm-blooded hosts (including livestock) and causes significant losses in the industry. In humans, it has been described as a pathogen in immunosuppressed people, it affects the fetus development in congenital infections, and is associated with various behavioral disorders in healthy people. Humans can acquire T. gondii by consuming undercooked, contaminated meat. Objective: To determine T. gondii positivity (currently unknown) in meat for human consumption (i.e., beef, chicken, and pork) in the city of Ibague, Colombia. Materials and methods: We used conventional nested PCR and the T. gondii B1 gene sequence as amplification target. We collected samples of meat (N=186) sold in the urban area of Ibagué (62 beef, 62 chicken, and 62 pork samples) and determined the T. gondii positivity percentage for each type of meat. Results: The study found an average of 18.8% positivity for all meat samples, pork having the highest percentage (22.5%; 14/62), followed by beef (19.3%; 12/62) and chicken (14.5%; 9/62). The best-amplified products were sequenced by macrogen and aligned with the B1 gene sequences in GenBank, thereby confirming their identity. Conclusions: This is the first study of T. gondii prevalence in meat for human consumption carried out in the city of Ibagué and the department of Tolima. All three types of meat sampled represent a risk for local human infection

Introducción. Toxoplasma gondii es un parásito con gran potencial zoonótico que puede infectar un amplio rango de huéspedes de sangre caliente, incluidos los animales del sector pecuario, lo que causa pérdidas a la industria. En el humano, es patógeno en personas inmunosuprimidas y afecta el desarrollo del feto en infecciones congénitas. Además, se asocia con diversos trastornos del comportamiento en personas sanas. El humano puede adquirir T. gondii al consumir carnes contaminadas mal cocidas. Objetivo. Determinar la positividad de T. gondii en carnes de consumo humano (res, pollo y cerdo) en Ibagué, Colombia. Materiales y métodos. Se utilizó la PCR convencional anidada y la secuencia del gen B1 de T. gondii como blanco de amplificación. Se tomaron 186 muestras de carne comercializada en la zona urbana de Ibagué (62 de res, 62 de pollo y 62 de cerdo) y se obtuvo el porcentaje de positividad en cada tipo de carne evaluada. Resultados. Se encontró un porcentaje de positividad de 18,8 % en las muestras, siendo la carne de cerdo la del mayor porcentaje (22,5 %; 14/62), seguida por las muestras de carne de res (19,3 %; 12/62) y de pollo (14,5 %; 9/62). Los mejores productos amplificados fueron secuenciados en Macrogen, y alineados con las secuencias del gen B1 depositadas en el GenBank, con lo que se confirmó su identidad. Conclusiones. Este es el primer estudio sobre prevalencia de T. gondii en carnes para consumo humano en Ibagué y el departamento del Tolima. Se demostró que los tres tipos de carne representan un riesgo para la infección en humanos a nivel local.

Maxicircle architecture and evolutionary insights into Trypanosoma cruzi complex.

We sequenced maxicircles from T. cruzi strains representative of the species evolutionary diversity by using long-read sequencing, which allowed us to uncollapse their repetitive regions, finding that their real lengths range from 35 to 50 kb. T. cruzi maxicircles have a common architecture composed of four regions: coding region (CR), AT-rich region, short (SR) and long repeats (LR). Distribution of genes, both in order and in strand orientation are conserved, being the main differences the presence of deletions affecting genes coding for NADH dehydrogenase subunits, reinforcing biochemical findings that indicate that complex I is not functional in T. cruzi. Moreover, the presence of complete minicircles into maxicircles of some strains lead us to think about the origin of minicircles. Finally, a careful phylogenetic analysis was conducted using coding regions of maxicircles from up to 29 strains, and 1108 single copy nuclear genes from all of the DTUs, clearly establishing that taxonomically T. cruzi is a complex of species composed by group 1 that contains clades A (TcI), B (TcIII) and D (TcIV), and group 2 (1 and 2 do not coincide with groups I and II described decades ago) containing clade C (TcII), being all hybrid strains of the BC type. Three variants of maxicircles exist in T. cruzi: a, b and c, in correspondence with clades A, B, and C from mitochondrial phylogenies. While A and C carry maxicircles a and c respectively, both clades B and D carry b maxicircle variant; hybrid strains also carry the b- variant. We then propose a new nomenclature that is self-descriptive and makes use of both the phylogenetic relationships and the maxicircle variants present in T. cruzi.

Salivary proteins electrophoretic patterns enabled differentiating Colombian Rhodnius Trans-Andean and Cis-Andean groups / [Los patrones electroforéticos de proteínas salivales permiten diferenciar los grupos transandino y cisandino de las especies de Rhodnius de Colombia].

Introduction: Rhodnius (Hemiptera: Reduviidae: Triatominae) species are made up of haematophagous insect vectors for Trypanosoma cruzi (Chagas' disease aetiological agent) and T. rangeli, an infective parasite that is not pathogenic for vertebrate hosts. The study of their salivary protein diversity enables the obtention of characteristic one-dimensional electrophoretic profiles of some triatomine species; however, few reports have dealt with Rhodnius species salivary proteins electrophoretic patterns. Objective: To compare R. colombiensis, R. pallescens, R. pictipes, R. prolixus, and R. robustus' salivary proteins one-dimensional electrophoretic profiles. Materials and methods: SDS-PAGE was used for obtaining electrophoretic profiles of saliva from the species under study. The unweighted pair group method with arithmetic mean (UPGMA) was used for constructing a phenogram. Results: Electrophoretic profiles of soluble saliva had protein bands ranging from 15 to 45 kDa, thereby enabling the five species studied to be differentiated. The phenogram revealed two main groups, one formed by the Pictipes and Prolixus cis-Andean groups and another consisting of the Pallescens trans-Andean group. Conclusion: Differences were revealed regarding R. colombiensis, R. pallescens, R. pictipes, R. prolixus, and R. robustus electrophoretic profiles of salivary proteins; their variability facilitated constructing a phenogram which was taxonomically congruent with the groups from the genus Rhodnius.

Introducción. Las especies Rhodnius (Hemiptera: Reduviidae: Triatominae) están conformadas por insectos hematófagos vectores de Trypanosoma cruzi, agente etiológico de la enfermedad de Chagas, y T. rangeli, parásito infectivo pero no patógeno para el vertebrado. El estudio de la diversidad proteica de la saliva de estos insectos permite la obtención de perfiles electroforéticos unidimensionales característicos de algunas especies de triatominos. Sin embargo, el reporte de los patrones electroforéticos de proteínas salivales de las especies de Rhodnius ha sido escaso. Objetivo. Hacer un análisis comparativo de los perfiles electroforéticos unidimensionales de las proteínas salivales de R. colombiensis, R. pallescens, R. pictipes, R. prolixus y R. robustus. Materiales y métodos. Se obtuvieron los perfiles electroforéticos de la saliva de las especies en estudio mediante electroforesis en gel de poliacrilamida con dodecilsulfatosódico (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis, SDS-PAGE) y se construyó un fenograma mediante el método UPGMA (Unweighted Pair Group Method Using Arithmetic Averages). Resultados. Los perfiles electroforéticos de las proteínas solubles de saliva presentaron bandas en un rango de masa aproximado de 15 a 45 kDa, los cuales permitieron diferenciar las cinco especies estudiadas. El fenograma reveló la existencia de dos grupos principales: uno conformado por los grupos cisandinos Pictipes y Prolixus y otro constituido por el grupo transandino Pallescens. Conclusiones. Existen diferencias en los perfiles electroforéticos de las proteínas salivales entre R. colombiensis, R. pallescens, R. pictipes, R. prolixus y R. robustus, cuya variabilidad permitió construir un fenograma congruente con los grupos del género Rhodnius.

The first molecular detection of Blastocystis subtypes in human faecal samples from Ibague, Colombia.

Blastocystis spp. has become one of the protozoans arousing the greatest scientific interest because of the controversy surrounding its biology; it is currently considered one of the most prevalent organisms in humans and animals worldwide. Such prevalence increases, especially in tropical countries where infection rates are high, highlighting the need to conduct studies focused on understanding this protozoan's biology. Interestingly, molecular tools are emerging as the best option for diagnosing this infection. This study was thus aimed at conventional PCR molecular detection and characterisation of Blastocystis spp. in human faecal samples from Ibagué, Colombia, using primers targeting the small subunit ribosomal ribonucleic acid (rRNA) gene. One hundred human faecal samples with confirmed Blastocystis spp. were studied, revealing the following subtype genetic diversity: ST1 50%, ST2 33% and ST3 17%. The results contributed to the limited information available regarding Blastocystis spp. in Colombia and created a reference point for further studies in the region.

ITS-RFLP- and RAPD-based genetic variability of Trypanosoma cruzi I, human and vector strains in Santander, Colombia.

Chagas disease is a severe public health problem in Latin-American countries. In Colombia, the predominance of Trypanosoma cruzi I has been described in the literature, with a broad heterogeneity between strains. However, most of the studies carried out centered on isoenzyme analysis, with a smaller number that focus on other molecular methods. In this report, we discuss the results of a molecular analysis of T. cruzi I strains, isolated from the domestic cycle, from the department of Santander, one of the territorial divisions where the prevalence of infection is highest. Internal transcribed spacer-restriction fragment length polymorphism and random amplification of polymorphic DNA were used to characterize 16 strains from human and vector (Triatominae) hosts. The data reveal a clustering based on the biological origin. Human and vector strains grouped separately; however, three vector strains clustered together with human strains. These results indicate that genetic differences exist in the strains that infect both hosts. We conclude that T. cruzi I populations in the domestic cycle of transmission of Chagas disease in Santander are heterogeneous and are composed of different clones. The epidemiological and biological implications are discussed.

Distribución de Aedes albopictus en Ibagué: potencial riesgo de brotes de arbovirosis

Introducción. Aedes albopictus es un vector de arbovirus como dengue, Zika, chikungunya y fiebre amarilla. Los primeros reportes en el continente americano datan de 1985 y dada su capacidad de adaptación ecológica y fisiológica, se ha distribuido rápidamente en el territorio colombiano desde su primer reporte en 1998. Objetivo. Determinar la distribución de A. albopictus en las comunas de Ibagué, Colombia. Materiales y métodos. Los muestreos se realizaron entre mayo y noviembre de 2022 en zonas con abundante vegetación de las 13 comunas de Ibagué. Se emplearon aspiradores y redes entomológicas. Los mosquitos fueron transportados al Laboratorio de Investigaciones en Parasitología Tropical de la Universidad del Tolima para su determinación taxonómica. Resultados. Se identificaron 708 ejemplares de A. albopictus, distribuidos en las comunas de Ibagué. La mayor abundancia del vector se presentó en las comunas 10, 11, 7, 8, 2 y 9. Las comunas 3, 4, 5, 6, 12 y 13 presentaron abundancias relativas cercanas al 3 %, y la comuna 1 tuvo una abundancia del 2 %. Conclusiones. Aedes albopictus está distribuido en todas las comunas de Ibagué, probablemente su dispersión se ha visto favorecida por las condiciones ambientales y sociales de esta región. Se recomienda hacer seguimiento anual a las poblaciones de este vector y realizar una caracterización molecular de los arbovirus encontrados. Además, el conocer la distribución de este mosquito en la ciudad permitirá focalizar las estrategias de control entomológico y prevenir futuros brotes de arbovirosis.

Introduction. Aedes albopictus is a vector for arboviruses, such as dengue, Zika, chikungunya, and yellow fever. The first A. albopictus reports on the American continent date back to 1985. It has spread rapidly throughout Colombia since its first report in 1998 due to its ecological and physiological adaptation capability. Objective. To determine A. albopictus distribution in the 13 communes of Ibagué, Colombia. Materials and methods. Samples were collected between May and November 2022 in the 13 communes of Ibagué. Vacuum sampling and sweep-netting entomological nets were used in areas with abundant vegetation. The mosquitoes were transported to the Laboratorio de Investigaciones en Parasitología Tropical at the Universidad del Tolima for taxonomic determination. Results. We identified 708 A. albopictus specimens distributed throughout Ibague's 13 communes. The highest vector abundance occurred in communes 10, 11, 7, 8, 2, and 9; communes 3, 4, 5, 6, 12, and 13 had a relative abundance of around 3%, while commune 1 had 2% of relative abundance. Conclusions. Aedes albopictus is distributed throughout all the communes of Ibague. Its dispersion has probably been favored by this region's environmental and social conditions. We recommend annual monitoring of these vectors populations and molecular characterization of the found arboviruses. Ascertaining this mosquito's distribution throughout the city will enable focusing entomological control strategies and preventing future arbovirus outbreaks.

Identifying four Trypanosoma cruzi I isolate haplotypes from different geographic regions in Colombia.

Trypanosoma cruzi has been classified into the groups T. cruzi I and T. cruzi II. The latter is subdivided into five smaller lineages based on multilocus enzyme electrophoresis and random amplified polymorphic DNA, designated as IIa-IIe, which shows correspondence with rRNA/mini-exon lineages. Twelve previously characterised T. cruzi isolates from different hosts, including humans, Didelphis marsupialis, and triatomines were analysed to establish genetic variability in T. cruzi group T. cruzi I isolates from different geographical regions of Colombia. DNA samples were sequenced based on the mini-exon gene intergenic region. Sequences were analysed using Clustal W, Staden 1.5 and MEGA3 software, and using reported sequences from the GenBank as reference. The genetic distances were analysed using Kimura's two-parameter model. The isolates' joint alignment was of 350bp, and the calculated nucleotide divergence was of 17.5%. The differences consisted of 23 transitions (7.2%), 14 transversions (4.4%) and 19 insertion-deletions (5.9%). The Colombian T cruzi I isolates revealed sufficient genetic variability for us to propose the existence of four haplotypes identified through single nucleotide polymorphism (SNP) and insertion/deletion found in the mini-exon gene's non-transcribed spacer intergenic region.

Detection of Trypanosoma cruzi and Trypanosoma rangeli infection in triatomine vectors by amplification of the histone H2A/SIRE and the sno-RNA-C11 genes.

Trypanosoma rangeli is non pathogenic for humans but of important medical and epidemiological interest because it shares vertebrate hosts, insect vectors, reservoirs and geographic areas with T. cruzi, the etiological agent of Chagas disease. Therefore, in this work, we set up two PCR reactions, TcH2AF/R and TrFR2, to distinguish T. cruzi from T. rangeli in mixed infections of vectors based on amplification of the histone H2A/SIRE and the small nucleolar RNA Cl1 genes, respectively. Both PCRs were able to appropriately detect all T. cruzi or T. rangeli experimentally infected-triatomines, as well as the S35/S36 PCR which amplifies the variable region of minicircle kDNA of T. cruzi. In mixed infections, whereas T. cruzi DNA was amplified in 100% of samples with TcH2AF/R and S35/S36 PCRs, T. rangeli was detected in 71% with TrF/R2 and in 6% with S35/S36. In a group of Rhodnius colombiensis collected from Coyaima (Colombia), T. cruzi was identified in 100% with both PCRs and T. rangeli in 14% with TrF/R2 and 10% with S35/S36 PCR. These results show that TcH2AF/R and TrF/R2 PCRs which are capable of recognizing all T. cruzi and T. rangeli strains and lineages could be useful for diagnosis as well as for epidemiological field studies of T. cruzi and T. rangeli vector infections.

[Electrophoretic patterns of salivary hemeproteins (nitrophorines) of Rhodnius colombiensis and R. prolixus (Hemiptera, Reduviidae, Triatominae)]. / Patrones electroforéticos de hemoproteínas salivares (nitroforinas) de Rhodnius colombiensis y Rhodnius prolixus (Hemiptera, Reduviidae, Triatominae).

INTRODUCTION: Salivary hemeprotein electrophoresis in starch gels is a recent technique used for differentiation of Rhodnius species with great phenotypic similarity. Furthermore, populations of the same species can be differentiated from geographically separated locales. Of the 15 described Rhodnius species in Latin America, at least eight have been reported in Colombia. OBJECTIVE: To use the salivary hemeproteins electrophoresis for R. prolixus and R. colombiensis identification. These two species are phenotypically similar and have overlapping domestic and sylvatic cycles where they occur in the upper basin of the Magdalena river, Central Colombia. MATERIAL AND METHODS: The content of salivary glands of each insect was subjected to starch gel electrophoresis using glycine buffer, and the bands were revealed with 3,3',5,5'-tetramethylbenzidine. Band patterns were photographically recorded. RESULTS: Electrophoretic patterns of salivary hemeproteins of R. prolixus and R. colombiensis were able to unequivocally differentiate the two species. CONCLUSION: The usefulness of the starch gel technique for distinguishing between R. prolixus and R. colombiensis was demonstrated as an additional tool to the morphometric and molecular methods already in use for differentiation of these two species.

[Comparison of feeding and defecation patterns of Rhodnius colombiensis and Rhodnius prolixus(Hempitera, Reduviidae, Triatominae) under laboratory conditions]. / Comparación de los patrones de alimentaci6n y defecación de Rhodnius colombiensis y Rhodnius prolixus (hemiptera, Reduviidae, triatominae) en condiciones de laboratorio.

INTRODUCTION: Rhodnius colombiensis occasionally comes into human dwellings and consequently its role as an important potential vector in the transmission of American trypanosomiasis has been suggested. OBJECTIVE: The potential role of R. colombiensis as vector was defined by comparing the feeding and defecation patterns between R. colombiensis and R. prolixus, the main domiciliary vector of Trypanosoma cruzi in Colombia. MATERIALS AND METHODS. For each developmental stage of R. colombiensis and R. prolixus the following data were collected: (1) time of feeding initiation, (2) the time for reaching the repletion, (3) the number of interruptions and defecations during the feeding, (4) the time between the end of the feeding and the first defecation, (5) the number of defecations during 10, 60 and 95 minutes of observation after feeding, and (6) the quantity of blood ingested. RESULTS: The mean time of feeding initiation of the fifth instar nymphs, males and females, showed significant differences between the two species. The average of insects that defecated within 10 minutes after feeding was higher for each successive stage of R. prolixus and showed significant differences with Rhodnius colombiensis. In contrast, the mean weight of blood ingested by each stage of R. colombiensis and R. prolixus was significantly different between the N1, N2, N5 and females of these species. CONCLUSION: Rhodnius colombiensis produced fewer defecations than R. prolixus during feeding. A higher percentage of R. prolixus defecated within 10, 60 and 95 minutes after feeding. However, R. colombiensis remains a longer time in contact with the vertebrate host, thus raising the probability of its role in transmission considering its occasional entry to human dwellings and its higher prevalences of infection withT. cruzi and T. rangeli.

[Comparison of the life cycles of Rhodnius colombiensis Moreno, Jurberg & Galvão, 1999 and R. prolixus Stal, 1872 (Hemiptera, Reduviidae, Triatominae) under laboratory conditions]. / Comparación del ciclo de vida de Rhodnius colombiensis Moreno, Jurberg & Galvão, 1999 y Rhodnius prolixus Stal, 1872 (Hemiptera, Reduviidae, Triatominae) en condiciones de laboratorio.

INTRODUCTION: Rhodnius colombiensis is a sylvatic triatomine associated with wine palm trees (Attalea butyracea) in the high basin of the Magdalena river (Colombia). The frequent invasion of these vectors into human dwellings and the high prevalences of natural infection with Trypanosoma cruzi of these insects suggest an important role in the transmission of Chagas disease. OBJECTIVE: The length of the life cycles of R. colombiensis and R. prolixus under laboratory conditions were compared. MATERIALS AND METHODS: Ninety-two individuals for each species were studied. The mean duration time of each stage, the number of bloodmeals for each stage, the percentage of mortality, the cause of death, the mean of eggs laid by female, the number of fertile eggs and the longevity of adults were recorded. RESULTS: The mean duration time of all stages of R. colombiensis was higher than in R. prolixus, producing significant differences in the overall time from egg to adult. The mean of total eggs and fertile eggs showed significant differences, being higher in R. prolixus than in R. colombiensis. The total mortality was 31.5% for R. colombiensis and 6.5% for R. prolixus. The longevity of females was higher in R. prolixus. CONCLUSIONS: The stages of R. prolixus are of relatively short duration. In general, the nymphs take fewer bloodmeals than R. colombiensis, the adults take more bloodmeals and oviposit a larger number of fertile eggs, and females have a greater longevity. These parameters indicated that R. prolixus has superior reproductive success in comparison with R. colombiensis under the experimental conditions used. These new life cycle data of R. colombiensis will be useful for maintenance of laboratory colonies.

[Trypanosoma rangeli parasite-vector-vertebrate interactions and their relationship to the systematics and epidemiology of American trypanosomiasis]. / Interacción tripanosoma-vector-vertebrado y su relación con la sistemática y la epidemiología de la tripanosomiasis Americana.

INTRODUCTION: Trypanosoma rangeli is a species of trypanosome second to T. cruzi, that is infective to humans in Latin America. Variability in the biological, biochemical and molecular characteristics between different isolates isolates of this parasite have been recorded. OBJECTIVE: Morphological and molecular characteristics were recorded from strains of T. rangeli that were isolated from different species of Rhodnius and maintained in different vertebrate species. MATERIALS AND METHODS: Nineteen strains of T. rangeli were isolated from R. prolixus, R. pallescens and R. colombiensis in Colombia, R. ecuadoriensis in Peru and R. pallescens in Panama. Polymorphism of blood trypomastigotes in ICR mice was evaluated and pleomorphism of P53 strain of T. rangeli KP1(-) inoculated in mouse, marsupial and canine was studied. RAPD analysis (randomly amplified polymorphic DNA analysis) of 12 strains isolated from four species of Rhodnius was performed. RESULT: Based on the total length of blood trypomastigotes, three discrete groups were observed. The P53 strain showed significant differences in the size of blood trypomastigotes in mouse, marsupial and canine. RAPD analysis showed that the strains segregated into two branches corresponding to strains of T. rangeli KP1(+) and T. rangeli KP1(-). All strains of T. rangeli KP1(-) clustered according to the species of Rhodnius from which they were isolated. CONCLUSION: These data reveal, for the first time, a close association amongst T. rangeli strains and Rhodnius species, confirming that each species of Rhodnius transmits to vertebrate hosts a parasite population with clear phenotypic and genotypic differences. This is further evidence that supports the concept of clonal evolution of these parasites.

Identifying Trypanosoma cruzi discreet typing units in triatomines collected in different natural regions of Perú.

INTRODUCTION: Trypanosoma cruzi has been divided by international consensus into six discrete typing units (DTU): TcI, TcII, TcIII, TcIV, TcV y TcVI. The factors determining the dynamics of T. cruzi genotypes vector transmission of Chagas' disease in the different geographical regions of Perú are still unknown. OBJECTIVE: To detect and type T. cruzi DTUs from the faeces of seven species of triatomines (Panstrongylus chinai, P. geniculatus, P. herreri, Rhodnius robustus, R. pictipes, Triatoma carrioni and T. infestans) captured in eight departments from different natural regions of Perú. MATERIALS AND METHODS: We examined 197 insects for detecting trypanosomes. DNA was extracted from each insect intestinal contents and PCR amplification of kDNA, SL-IR, 24Sα rRNA and 18Sα RNA was performed for detecting T. cruzi DTUs. RESULTS: Five T. rangeli and 113 T. cruzi infections were detected; 95 of the latter were identified as TcI (two in P. chinai, one in P. geniculatus, 68 in P. herreri, four in R. pictipes, seven in R. robustus, one in T. carrioni, 12 in T. infestans), five as TcII (four in P. herreri, one in T. infestans), four as TcIII (three in P. herreri, one in R. robustus) and four TcIV infections in P. herreri. CONCLUSIONS: This is the first study which has attempted a large-scale characterization of T. cruzi found in the intestine of epidemiologically important vectors in Perú, thus providing basic information that will facilitate a better understanding of the dynamics of T. cruzi vector transmission in Perú.

Detección molecular de Toxoplasma gondii en carnes para consumo humano en Ibagué, Colombia / Molecular detection of Toxoplasma gondii in meat for human consumption in Ibague, Colombia

Introducción. Toxoplasma gondii es un parásito con gran potencial zoonótico que puede infectar un amplio rango de huéspedes de sangre caliente, incluidos los animales del sector pecuario, lo que causa pérdidas a la industria. En el humano, es patógeno en personas inmunosuprimidas y afecta el desarrollo del feto en infecciones congénitas. Además, se asocia con diversos trastornos del comportamiento en personas sanas. El humano puede adquirir T. gondii al consumir carnes contaminadas mal cocidas. Objetivo. Determinar la positividad de T. gondii en carnes de consumo humano (res, pollo y cerdo) en Ibagué, Colombia. Materiales y métodos. Se utilizó la PCR convencional anidada y la secuencia del gen B1 de T. gondii como blanco de amplificación. Se tomaron 186 muestras de carne comercializada en la zona urbana de Ibagué (62 de res, 62 de pollo y 62 de cerdo) y se obtuvo el porcentaje de positividad en cada tipo de carne evaluada. Resultados. Se encontró un porcentaje de positividad de 18,8 % en las muestras, siendo la carne de cerdo la del mayor porcentaje (22,5 %; 14/62), seguida por las muestras de carne de res (19,3 %; 12/62) y de pollo (14,5 %; 9/62). Los mejores productos amplificados fueron secuenciados en Macrogen, y alineados con las secuencias del gen B1 depositadas en el GenBank, con lo que se confirmó su identidad. Conclusiones. Este es el primer estudio sobre prevalencia de T. gondii en carnes para consumo humano en Ibagué y el departamento del Tolima. Se demostró que los tres tipos de carne representan un riesgo para la infección en humanos a nivel local.

Introduction: Toxoplasma gondii is a parasite with great zoonotic potential. It can infect a broad range of warm-blooded hosts (including livestock) and causes significant losses in the industry. In humans, it has been described as a pathogen in immunosuppressed people, it affects the fetus development in congenital infections, and is associated with various behavioral disorders in healthy people. Humans can acquire T. gondii by consuming undercooked, contaminated meat. Objective: To determine T. gondii positivity (currently unknown) in meat for human consumption (i.e., beef, chicken, and pork) in the city of Ibague, Colombia. Materials and methods: We used conventional nested PCR and the T. gondii B1 gene sequence as amplification target. We collected samples of meat (N=186) sold in the urban area of Ibagué (62 beef, 62 chicken, and 62 pork samples) and determined the T. gondii positivity percentage for each type of meat. Results: The study found an average of 18.8% positivity for all meat samples, pork having the highest percentage (22.5%; 14/62), followed by beef (19.3%; 12/62) and chicken (14.5%; 9/62). The best-amplified products were sequenced by macrogen and aligned with the B1 gene sequences in GenBank, thereby confirming their identity. Conclusions: This is the first study of T. gondii prevalence in meat for human consumption carried out in the city of Ibagué and the department of Tolima. All three types of meat sampled represent a risk for local human infection.

Systematic review on the biology, ecology, genetic diversity and parasite transmission potential of Panstrongylus geniculatus (Latreille 1811) in Latin America

Panstrongylus geniculatus (Latreille, 1811) is the triatomine with the largest geographic distribution in Latin America. It has been reported in 18 countries from southern Mexico to northern Argentina, including the Caribbean islands. Although most reports indicate that P. geniculatus has wild habitats, this species has intrusive habits regarding human dwellings mainly located in intermediate deforested areas. It is attracted by artificial light from urban and rural buildings, raising the risk of transmission of Trypanosoma cruzi. Despite the wide body of published information on P. geniculatus, many knowledge gaps exist about its biology and epidemiological potential. For this reason, we analysed the literature for P. geniculatus in Scopus, PubMed, Scielo, Google Scholar and the BibTriv3.0 databases to update existing knowledge and provide better information on its geographic distribution, life cycle, genetic diversity, evidence of intrusion and domiciliation, vector-related circulating discrete taxonomic units, possible role in oral T. cruzi transmission, and the effect of climate change on its biology and epidemiology.

Trypanosoma rangeli displays a clonal population structure, revealing a subdivision of KP1(-) strains and the ancestry of the Amazonian group.

Assessment of the genetic variability and population structure of Trypanosoma rangeli, a non-pathogenic American trypanosome, was carried out through microsatellite and single-nucleotide polymorphism analyses. Two approaches were used for microsatellite typing: data mining in expressed sequence tag /open reading frame expressed sequence tags libraries and PCR-based Isolation of Microsatellite Arrays from genomic libraries. All microsatellites found were evaluated for their abundance, frequency and usefulness as markers. Genotyping of T. rangeli strains and clones was performed for 18 loci amplified by PCR from expressed sequence tag/open reading frame expressed sequence tags libraries. The presence of single-nucleotide polymorphisms in the nuclear, multi-copy, spliced leader gene was assessed in 18 T. rangeli strains, and the results show that T. rangeli has a predominantly clonal population structure, allowing a robust phylogenetic analysis. Microsatellite typing revealed a subdivision of the KP1(-) genetic group, which may be influenced by geographical location and/or by the co-evolution of parasite and vectors occurring within the same geographical areas. The hypothesis of parasite-vector co-evolution was corroborated by single-nucleotide polymorphism analysis of the spliced leader gene. Taken together, the results suggest three T. rangeli groups: (i) the T. rangeli Amazonian group; (ii) the T. rangeli KP1(-) group; and (iii) the T. rangeli KP1(+) group. The latter two groups possibly evolved from the Amazonian group to produce KP1(+) and KP1(-) strains.