RESUMO
Colombia aims to eliminate malaria by 2030 but remains one of the highest burden countries in the Americas. Plasmodium vivax contributes half of all malaria cases, with its control challenged by relapsing parasitaemia, drug resistance and cross-border spread. Using 64 Colombian P. vivax genomes collected between 2013 and 2017, we explored diversity and selection in two major foci of transmission: Chocó and Córdoba. Open-access data from other countries were used for comparative assessment of drug resistance candidates and to assess cross-border spread. Across Colombia, polyclonal infections were infrequent (12%), and infection connectivity was relatively high (median IBD = 5%), consistent with low endemicity. Chocó exhibited a higher frequency of polyclonal infections (23%) than Córdoba (7%), although the difference was not significant (P = 0.300). Most Colombian infections carried double pvdhfr (95%) and single pvdhps (71%) mutants, but other drug resistance mutations were less prevalent (< 10%). There was no evidence of selection at the pvaat1 gene, whose P. falciparum orthologue has recently been implicated in chloroquine resistance. Global population comparisons identified other putative adaptations. Within the Americas, low-level connectivity was observed between Colombia and Peru, highlighting potential for cross-border spread. Our findings demonstrate the potential of molecular data to inform on infection spread and adaptation.
Assuntos
Antimaláricos , Malária Falciparum , Malária Vivax , Humanos , Plasmodium vivax/genética , Antimaláricos/farmacologia , Colômbia/epidemiologia , Malária Vivax/epidemiologia , Malária Vivax/tratamento farmacológico , Proteínas de Protozoários/genética , Resistência a Medicamentos/genética , GenômicaRESUMO
The human malaria parasite Plasmodium falciparum is globally widespread, but its prevalence varies significantly between and even within countries. Most population genetic studies in P. falciparum focus on regions of high transmission where parasite populations are large and genetically diverse, such as sub-Saharan Africa. Understanding population dynamics in low transmission settings, however, is of particular importance as these are often where drug resistance first evolves. Here, we use the Pacific Coast of Colombia and Ecuador as a model for understanding the population structure and evolution of Plasmodium parasites in small populations harboring less genetic diversity. The combination of low transmission and a high proportion of monoclonal infections means there are few outcrossing events and clonal lineages persist for long periods of time. Yet despite this, the population is evolutionarily labile and has successfully adapted to changes in drug regime. Using newly sequenced whole genomes, we measure relatedness between 166 parasites, calculated as identity by descent (IBD), and find 17 distinct but highly related clonal lineages, six of which have persisted in the region for at least a decade. This inbred population structure is captured in more detail with IBD than with other common population structure analyses like PCA, ADMIXTURE, and distance-based trees. We additionally use patterns of intra-chromosomal IBD and an analysis of haplotypic variation to explore past selection events in the region. Two genes associated with chloroquine resistance, crt and aat1, show evidence of hard selective sweeps, while selection appears soft and/or incomplete at three other key resistance loci (dhps, mdr1, and dhfr). Overall, this work highlights the strength of IBD analyses for studying parasite population structure and resistance evolution in regions of low transmission, and emphasizes that drug resistance can evolve and spread in small populations, as will occur in any region nearing malaria elimination.
Assuntos
Antimaláricos , Malária Falciparum , Parasitos , Animais , Humanos , Plasmodium falciparum/genética , Antimaláricos/farmacologia , Antimaláricos/uso terapêutico , Malária Falciparum/tratamento farmacológico , Malária Falciparum/epidemiologia , Malária Falciparum/parasitologia , Cloroquina/uso terapêutico , Resistência a Medicamentos/genética , América do Sul/epidemiologiaRESUMO
Characterising connectivity between geographically separated biological populations is a common goal in many fields. Recent approaches to understanding connectivity between malaria parasite populations, with implications for disease control efforts, have used estimates of relatedness based on identity-by-descent (IBD). However, uncertainty around estimated relatedness has not been accounted for. IBD-based relatedness estimates with uncertainty were computed for pairs of monoclonal Plasmodium falciparum samples collected from five cities on the Colombian-Pacific coast where long-term clonal propagation of P. falciparum is frequent. The cities include two official ports, Buenaventura and Tumaco, that are separated geographically but connected by frequent marine traffic. Fractions of highly-related sample pairs (whose classification using a threshold accounts for uncertainty) were greater within cities versus between. However, based on both highly-related fractions and on a threshold-free approach (Wasserstein distances between parasite populations) connectivity between Buenaventura and Tumaco was disproportionally high. Buenaventura-Tumaco connectivity was consistent with transmission events involving parasites from five clonal components (groups of statistically indistinguishable parasites identified under a graph theoretic framework). To conclude, P. falciparum population connectivity on the Colombian-Pacific coast abides by accessibility not isolation-by-distance, potentially implicating marine traffic in malaria transmission with opportunities for targeted intervention. Further investigations are required to test this hypothesis. For the first time in malaria epidemiology (and to our knowledge in ecological and epidemiological studies more generally), we account for uncertainty around estimated relatedness (an important consideration for studies that plan to use genotype versus whole genome sequence data to estimate IBD-based relatedness); we also use threshold-free methods to compare parasite populations and identify clonal components. Threshold-free methods are especially important in analyses of malaria parasites and other recombining organisms with mixed mating systems where thresholds do not have clear interpretation (e.g. due to clonal propagation) and thus undermine the cross-comparison of studies.
Assuntos
Genoma de Protozoário/genética , Malária Falciparum/parasitologia , Modelos Genéticos , Plasmodium falciparum/genética , Colômbia/epidemiologia , Frequência do Gene , Técnicas de Genotipagem , Humanos , Malária Falciparum/epidemiologia , Malária Falciparum/transmissão , Cadeias de Markov , Plasmodium falciparum/isolamento & purificação , Polimorfismo de Nucleotídeo Único , Reprodução Assexuada/genética , Análise Espaço-Temporal , IncertezaRESUMO
Self-medication with antimalarial drugs is a major factor in the development of drug resistance, exerting subtherapeutic drug pressure on circulating parasite populations. Data on self-medication with antimalarials from the Southern Pacific coast region of Colombia, where 4-aminoquinolines resistance and political instability prevail, are vital to elimination strategies. We present results of an exploratory study of 254 individuals having malaria symptoms who sought malaria diagnosis in two hospitals in Tumaco, Department of Nariño, Colombia. Thirty-two percent (82/254) of participants had positive Saker-Solomons urine tests, indicating self-medication with chloroquine (CQ) before consultation for diagnosis. Notably, among 30 pregnant women participating in the study, 43% were Saker--Solomons positive. Molecular analysis of the K76T position encoded by the pfcrt gene revealed the mutant allele in all four samples that were both positive for Plasmodium falciparum and positive for the Saker-Solomons test, suggesting persistent CQ pressure. The high frequency of self-medication, particularly among pregnant women merits attention by public health authorities and comprehensive investigation.
Assuntos
Antimaláricos/urina , Cloroquina/urina , Resistência a Múltiplos Medicamentos/genética , Malária Falciparum/tratamento farmacológico , Plasmodium falciparum/efeitos dos fármacos , Automedicação/estatística & dados numéricos , Adolescente , Adulto , Alelos , Antimaláricos/uso terapêutico , Cloroquina/uso terapêutico , Colômbia , Feminino , Humanos , Masculino , Proteínas de Membrana Transportadoras/genética , Pessoa de Meia-Idade , Mutação , Plasmodium falciparum/genética , Gravidez , Proteínas de Protozoários/genética , Adulto JovemRESUMO
BACKGROUND: Resistance to chloroquine and antifolate drugs has evolved independently in South America, suggesting that genotype - phenotype studies aimed at understanding the genetic basis of resistance to these and other drugs should be conducted in this continent. This research was conducted to better understand the population structure of Colombian Plasmodium falciparum in preparation for such studies. RESULTS: A set of 384 SNPs were genotyped in blood spot DNA samples from 447 P. falciparum infected subjects collected over a ten year period from four provinces of the Colombian Pacific coast to evaluate clonality, population structure and linkage disequilibrium (LD). Most infections (81%) contained a single predominant clone. These clustered into 136 multilocus genotypes (MLGs), with 32% of MLGs recovered from multiple (2 - 28) independent subjects. We observed extremely low genotypic richness (R = 0.42) and long persistence of MLGs through time (median = 537 days, range = 1 - 2,997 days). There was a high probability (>5%) of sampling parasites from the same MLG in different subjects within 28 days, suggesting caution is needed when using genotyping methods to assess treatment success in clinical drug trials. Panmixia was rejected as four well differentiated subpopulations (FST = 0.084 - 0.279) were identified. These occurred sympatrically but varied in frequency within the four provinces. Linkage disequilibrium (LD) decayed more rapidly (r2 = 0.17 for markers <10 kb apart) than observed previously in South American samples. CONCLUSIONS: We conclude that Colombian populations have several advantages for association studies, because multiple clone infections are uncommon and LD decays over the scale of one or a few genes. However, the extensive population structure and low genotype richness will need to be accounted for when designing and analyzing association studies.
Assuntos
Plasmodium falciparum/genética , Colômbia , Genética Populacional , Humanos , Desequilíbrio de Ligação , Malária/epidemiologia , Polimorfismo de Nucleotídeo ÚnicoRESUMO
OBJECTIVE: To evaluate the dye extent and distribution at the lumbar plexus (LP) of three volumes of local anaesthetic-methylene-blue solution administered close to the femoral nerve (FN) by the use of a ventral ultrasound (US)-guided suprainguinal approach (SIA). STUDY DESIGN: Prospective experimental trial. ANIMALS: Twenty mongrel canine cadavers weighing 17.7 ± 3.8 kg (mean ± SD). METHODS: The left and right LP of two cadavers were dissected to identify the FN, obturator nerve (ON) and lateral femoral cutaneous nerve (LFCN). The extent and distribution of dye at the LP of each of three volumes of injectate of 0.2, 0.4 and 0.6 mL kg(-1) administered close to the FN by a ventral US-guided SIA then were studied in a further 18 dog cadavers (n = 6 per group). Staining of ≥2 cm along the target nerves was indicative of sufficient spread to produce a nerve block. RESULTS: The ventral US-guided SIA allowed the observation of the FN within the iliopsoas muscle (IPM) in a total of 17 cadavers. The assessment of the dye extent and distribution revealed a similar pattern regardless of the injected volume. From the injection site, the spreading of injectate occurred in cranial, lateral and caudal directions. The FN and ON were effectively stained in all the cases. The LFCN was not effectively stained in any case. CONCLUSIONS AND CLINICAL RELEVANCE: A volume of 0.2 mL kg(-1) administered close to the FN by a ventral US-guided SIA produced a sufficient distribution of the injectate within the IPM to produce effective staining of the FN and ON. This US-guided technique may be an appropriate alternative to previously reported techniques based on electrolocation to block the FN and ON in the dog.
Assuntos
Cães , Nervo Femoral/anatomia & histologia , Bloqueio Nervoso/veterinária , Nervo Obturador/anatomia & histologia , Ultrassonografia de Intervenção/veterinária , Anestésicos Locais/administração & dosagem , Animais , Cadáver , Lidocaína/administração & dosagem , Bloqueio Nervoso/métodos , Ultrassonografia de Intervenção/métodosRESUMO
This prospective study assessed a ventral ultrasound-guided suprainguinal approach to block the femoral nerve (FN) in dogs. The anatomical features of the FN were evaluated in four canine cadavers. In another five cadavers, the FN was located by ultrasound-guidance and the accuracy of this technique was evaluated by injection of black ink and posterior evaluation of the degree of staining of the nerves. In five live dogs, the FN was blocked with 2% lidocaine. The distribution of lidocaine around the nerve and the presence of motor deficit were evaluated. The FN was easily located and accurately blocked in all cases. This new ultrasound-guided approach was reliable for blocking the FN and might be a suitable alternative to the traditional approaches described to block the FN in the dog.
Assuntos
Anestésicos Locais/administração & dosagem , Nervo Femoral/anatomia & histologia , Lidocaína/administração & dosagem , Bloqueio Nervoso/veterinária , Ultrassonografia de Intervenção/veterinária , Animais , Cadáver , Cães , Bloqueio Nervoso/métodosRESUMO
Drug resistance is one of the principal obstacles blocking worldwide malaria control. In Colombia, malaria remains a major public health concern and drug-resistant parasites have been reported. In vitro drug susceptibility assays are a useful tool for monitoring the emergence and spread of drug-resistant Plasmodium falciparum. The present study was conducted as a proof of concept for an antimalarial drug resistance surveillance network based on in vitro susceptibility testing in Colombia. Sentinel laboratories were set up in three malaria endemic areas. The enzyme linked immunosorbent assay-histidine rich protein 2 and schizont maturation methods were used to assess the susceptibility of fresh P. falciparum isolates to six antimalarial drugs. This study demonstrates that an antimalarial drug resistance surveillance network based on in vitro methods is feasible in the field with the participation of a research institute, local health institutions and universities. It could also serve as a model for a regional surveillance network. Preliminary susceptibility results showed widespread chloroquine resistance, which was consistent with previous reports for the Pacific region. However, high susceptibility to dihydroartemisinin and lumefantrine compounds, currently used for treatment in the country, was also reported. The implementation process identified critical points and opportunities for the improvement of network sustainability strategies.
Assuntos
Antimaláricos/farmacologia , Resistência a Medicamentos , Plasmodium falciparum/efeitos dos fármacos , Colômbia , Humanos , Concentração Inibidora 50 , Malária Falciparum/parasitologia , Testes de Sensibilidade Parasitária/métodosRESUMO
Drug resistance is one of the principal obstacles blocking worldwide malaria control. In Colombia, malaria remains a major public health concern and drug-resistant parasites have been reported. In vitro drug susceptibility assays are a useful tool for monitoring the emergence and spread of drug-resistant Plasmodium falciparum. The present study was conducted as a proof of concept for an antimalarial drug resistance surveillance network based on in vitro susceptibility testing in Colombia. Sentinel laboratories were set up in three malaria endemic areas. The enzyme linked immunosorbent assay-histidine rich protein 2 and schizont maturation methods were used to assess the susceptibility of fresh P. falciparum isolates to six antimalarial drugs. This study demonstrates that an antimalarial drug resistance surveillance network based on in vitro methods is feasible in the field with the participation of a research institute, local health institutions and universities. It could also serve as a model for a regional surveillance network. Preliminary susceptibility results showed widespread chloroquine resistance, which was consistent with previous reports for the Pacific region. However, high susceptibility to dihydroartemisinin and lumefantrine compounds, currently used for treatment in the country, was also reported. The implementation process identified critical points and opportunities for the improvement of network sustainability strategies.
Assuntos
Humanos , Antimaláricos , Resistência a Medicamentos , Plasmodium falciparum , Colômbia , Malária Falciparum , Testes de Sensibilidade Parasitária/métodosRESUMO
Most rapid diagnostic tests (RDTs) available use histidine-rich protein 2 (HRP2) as a target. However, it has been reported that sequence variations of this protein affects its sensitivity. Currently, there is insufficient evidence for HRP2 variability in Plasmodium falciparum isolates from Colombia and its relationship with RDT performance. To determine possible geographic differences and their effects on the performance of RDTs, 22 blood samples from patients with P. falciparum malaria from Tumaco and Buenaventura, Colombia were assessed by measurement of HRP2 concentration by an HRP2 enzyme-linked immunosorbent assay, RDTs, and thick blood smear. Statistical analysis showed an association between RDT performance and HRP2 concentrations. No significant difference was found between locations. A large variation of antigen concentration in samples was found at same parasitemia. In contrast to previously reports, there was no correlation between initial parasitemia and HRP2 concentration. Our results indicate that antigen quantity should be studied more carefully because the sensitivity of the RDT is affected more by antigen concentration than by parasitemia.