Caracterización molecular de parásitos del género Leishmania en lesiones cutáneas con frotis y cultivo negativos

Entre las zoonosis parasitarias endémicas en Panamá, la Leishmaniasis Cutánea (LC) presenta las incidencias más altas con 1,000-3,000 casos anuales. El diagnóstico parasitológico de esta infección se basa en la demostración microscópica del parásito en frotis teñidos y cultivos realizados a partir de las lesiones. Sin embargo, con frecuencia estos análisis resultan negativos aun cuando las pruebas moleculares (PCR) confirman posteriormente la infección. El objetivo principal de este estudio fue detectar y caracterizar molecularmente los parásitos del género Leishmania presentes en lesiones cutáneas con frotis y cultivo negativos. Para ello se evaluaron 123 muestras de ADN procedentes de lesiones de LC de pacientes que asistieron a la Unidad de Diagnóstico, Investigación Clínica y Medicina Tropical del ICGES entre el período del 2015 al 2019. Estos casos presentaron diagnósticos directos negativos, pero fueron confirmados por alguna metodología molecular. Las muestras fueron evaluadas inicialmente con 4 técnicas de PCR que amplifican las siguientes regiones: (1) KDNA-Viannia específico, (2) KDNA-Género específico, (3) Hsp70-Género específico y (4) qPCR 18S ribosomal. Los hallazgos confirman la mayor sensibilidad de la PCR KDNA-Viannia específico (100%). Los resultados de la qPCR18S ribosomal, mostraron una sensibilidad de 77.6%. Adicionalmente, 69 muestras positivas en el PCR-Hsp-70 fueron caracterizadas a nivel de especie mediante un análisis RFLP con las enzimas Hae III y Bcc 1; correspondiendo un 59.4% a L. (y.) panamensis y 40.6% a L. (y.) guyanensis. Finalmente, los productos de amplificación del PCR-Hsp-70 fueron purificados y secuenciados para un análisis filogenético que incluyó 38 secuencias. Veintiuna de las secuencias evaluadas se agruparon con una secuencia referencia de L. (y.) panamensis, mientras que 11 caracterizadas por RFLP como L. (y.) guyanensis se agruparon con el grupo de la variante genética denominada Leishmania sp.1. Según nuestros resultados, en estas lesiones de LO con baja carga parasitaria predomina L. (y.) panamensis, sin embargo, la variante genética Leishmania sp.1 también se vincula con frecuencia con este tipo de lesiones. En ningún caso se logró demostrar una asociación significativa entre estas lesiones con baja carga parasitaria y otra característica clínico-epidemiológica. Los resultados de este estudio refuerzan la utilidad de los métodos moleculares en el diagnóstico de la LO, especialmente en lesiones con baja carga parasitaria. Además, señalan las especies/variantes genéticas de Leishmania involucradas, así como las características clínico/epidemiológicas vinculadas con este tipo de lesión

Sexual and reproductive health and access : Results of a rapid epidemiological assessment among migrant peoples in transit through Darién, Panamá

Background: The foot transit of migrant peoples originating from the Caribbean, South America, Asia, and Sub-Saharan Africa through the Darién Forest (DF) in Eastern Panamá towards North America has increased in recent years from approximately 30,000 people/year to >133,000 in 2021. In the DF, there is no food/housing provision nor healthcare access. Very little is known of sexual and reproductive health (SRH) among this population. This study used rapid epidemiological methods to describe the SRH situation among migrant peoples in transit through the DF. Methods: This cross-sectional study randomly selected migrant people in transit (men and women) at a Migrant Reception Station in Darién, Panamá, between January 4-11, 2022. Data collection included a self-applied questionnaire (≥18 years); clinical screening (≥12 years); and HCG, treponemal antibodies, and HIV(I/II) lateral-flow tests with blood samples (≥12 years). Descriptive analyses were used to report findings. Results: In all, 69 men and 55 women participated in the self-applied questionnaire, 70 men and 51 women in clinical screening; 78 men and 63 women in HCG, treponemal antibody and HIV testing. Overall, 26.1% (18/69) men and 36.4% (20/55) women reported sexual intercourse within the past month. The last sex partner was casual among 43.0% (21/49) of men and 27.8% (10/36) of women; of those, 42.9% (9/21) of men and 80.0% (8/10) of women reported this sex was condomless. Among women, 20.0% (11/55) tested positive for pregnancy; 5 of these pregnancies were planned. Of those screened, a reproductive tract infection symptom was reported by 5.7% (4/70) of men and 58.8% (30/51) of women. A total of 32.7% (18/55) of men and 18.2% (8/44) of women reported no prior HIV testing. Of 78 men, HIV and treponemal antibodies were found among 1.3% (n = 1) and 2.6% (n = 2), and among 63 women, 3.2% (n = 2) and 3.2% (n = 2), respectively. Conclusions: This rapid epidemiological assessment found high recent sexual activity, low condom use with casual partners, and a need for increased HIV and syphilis testing and treatment. There is a need for increased testing, condom provision, and SRH healthcare access at migrant reception stations that receive migrant peoples in transit through Panamá

Characterization of <i>Leishmania</i> spp. Causing Cutaneous Lesions with a Negative Parasitological Diagnosis in Panama

A total of 123 DNA samples from Panamanian patients with cutaneous leishmaniasis (CL) lesions were evaluated. These samples were previously confirmed with CL by a specific KDNA-Viannia PCR but had a negative parasitological diagnosis (Group A). Epidemiological variables, such as age, sex, geographic origin, evolution time, and the number and location of the lesions, were analyzed. No significant differences (p Leishmania (Viannia) panamensis and 40.6% (28/69) to Leishmania (Viannia) guyanensis. Finally, the sequence and phylogenetic analysis of 32 of the samples confirmed the species in 21 (65.6%, 21/32) samples, originally characterized as L. (V.) panamensis. However, 11 samples (34.4%, 11/32), initially identified via RFLP-Hsp70 as L. (V.) guyanensis, matched the sequence of a genetic variant known as Leishmania sp.1. These results point out the species/genetic variants of Leishmania in the case of CL lesions with an apparently low parasite load

U.S. bound journey of migrant peoples InTransit across Dante's Inferno and Purgatory in the Americas

Rapid rise of population migration is a defining feature of the 21st century due to the impact of climate change, political instability, and socioeconomic downturn. Over the last decade, an increasing number of migrant peoples travel across the Americas to reach the United States seeking asylum or cross the border undocumented in search of economic opportunities. In this journey, migrant people experience violations of their human rights, hunger, illness, violence and have limited access to medical care. In the ‘Divine Comedy’, the Italian poet Dante Alighieri depicts his allegorical pilgrimage across Hell and Purgatory to reach Paradise. More than 700 years after its publication, Dante's poem speaks to the present time and the perilious journey of migrant peoples to reach safehavens. By exploring the depths and heights of the human condition, Dante's struggles resonate with the multiple barriers and the unfathomable experiences faced by migrant peoples in transit across South, Central, and North America to reach the United States. Ensuring the safety of migrant peoples across the Americas and elsewhere, and attending to their health needs during their migratory paths represent modern priorities to reduce social injustices and achieving health equity.Fund Neglected Tropical Diseases of the University of Colorado by Mr. Howard Janzen