Characterisation of an area of coexistent visceral and cutaneous leishmaniasis transmission in the State of Piauí, Brazil

BACKGROUND In Brazil, transmission of visceral and cutaneous leishmaniasis has expanded geographically over the last decades, with both clinical forms occurring simultaneously in the same area. OBJECTIVES This study characterised the clinical, spatial, and temporal distribution, and performed entomological surveillance and natural infection analysis of a leishmaniasis-endemic area. METHODS In order to characterise the risk of leishmaniasis transmission in Altos, Piauí, we described the clinical and socio-demographic variables and the spatial and temporal distribution of cases of American visceral leishmaniasis (AVL) and American cutaneous leishmaniasis (ACL) cases and identified potential phlebotomine vectors. FINDINGS The urban area concentrated almost 54% of ACL and 86.8% of AVL cases. The temporal and spatial distribution of AVL and ACL cases in Altos show a reduction in the number of risk areas, but the presence of permanent disease transmission foci is observed especially in the urban area. 3,808 phlebotomine specimens were captured, with Lutzomyia longipalpis as the most frequent species (98.45%). Of the 35 females assessed for natural infection, one specimen of Lu. longipalpis tested positive for the presence of Leishmania infantum and Leishmania braziliensis DNA. MAIN CONCLUSION Our results indicate the presence of risk areas for ACL and AVL in the municipality of Altos and highlight the importance of entomological surveillance to further understand a possible role of Lu. longipalpis in ACL transmission.

Viable Leishmania parasites in the absence of an in vitro IFN-γ response in asymptomatic carriers

ABSTRACT Asymptomatic infection (the absence or inapparent signs and symptoms) has been observed in many endemic areas of leishmaniasis, however, little is known about the parasitological and immunological factors associated with this type of infection. This study aimed to identify the in vitro expression of IFN-γ in asymptomatic carriers of viable Leishmania parasites. Asymptomatic infection was identified using the Montenegro skin test in an at-risk population from Yucatan, Mexico. Parasite viability was evinced in the blood by 7SL RNA transcripts amplification. The expression of mRNA IFN-γ was analyzed in peripheral blood mononuclear cells stimulated with soluble Leishmania antigen, using RT-qPCR. Parasite viability was observed in 33.3 % (5/15) of asymptomatic subjects. No differences were found in the expression of IFN-γ between asymptomatic and healthy subjects, and no correlation was found between the presence of viable parasites and the expression of IFN-γ. This study demonstrates the persistence of Leishmania parasites in the absence of an in vitro IFN-γ response in asymptomatic carriers from Mexico.

Perfil epidemiológico das notificações de leishmaniose no estado de Pernambuco no Brasil de 2015-2019 / Epidemiological profile of leishmaniasis notifications in the Pernambuco state in Brazil from 2015-2019

Aqui, fornecemos uma análise epidemiológica da leishmaniose em Pernambuco, Brasil, de 2015 a 2019. Nossos resultados revelaram 1.705 casos de leishmaniose tegumentar americana-LTA e 859 casos de leishmaniose visceral-LV. LV e LTA afetam crianças de 1 a 4 anos (23% e 3,2% dos casos, respectivamente) e adultos de 20 a 39 anos (26% e 28%, respectivamente), principalmente mulheres. O maior percentual (54,6%) de casos de LV foi encontrado na área urbana, enquanto LTA ocorreu principalmente na área rural (63,2%). Indivíduos com maior escolaridade foram menos acometidos pela leishmaniose e dados de prognóstico revelaram boas chances de cura clínica, mas óbitos ainda são registrados. Esses dados mostram que ações preventivas de saúde devem ser desenvolvidas para impedir a disseminação da leishmaniose, reduzir seu impacto na saúde pública e restaurar a qualidade de vida da população. Este estudo pode ser útil na elaboração de estratégias para minimizar os impactos da leishmaniose.

Here, we provide an epidemiological analysis of leishmaniasis in Pernambuco, Brazil, from 2015 to 2019. Our results revealed 1,705 of American cutaneous leishmaniasis-ACL and 859 cases of visceral leishmaniasis-VL. VL and ACL affect children aged 1­4 years (23% and 3.2% of all cases, respectively) and adults aged 20­39 years (26% and 28%, respectively), mainly women. The highest percentage (54.6%) of VL cases was found in urban areas, while ACL occurred mainly in rural areas (63.2%). Individuals with a higher level of education were less affected by leishmaniasis and data on prognosis revealed good chances of clinical cure, but deaths are still recorded. These data show that preventive health actions must be developed to hinder leishmaniasis spread, reduce its impact on public health, and restore the population's life quality. This study can be useful in further elaboration of strategies to minimize leishmaniasis impacts.

Comparación de la eficacia y toxicidad del estibogluconato de sodio y antimoniato de meglumina en el tratamiento de leishmaniasis cutánea en Perú / Efficacy and safety of sodium stibogluconate compared to meglumine antimoniate in the treatment of cutaneous leishmaniasis in Peru

Objetivos : Comparar la eficacia y toxicidad del antimoniato de meglumina (AM) y estibogluconato sódico (EGS) en el tratamiento de leishmaniasis cutánea (LC) en un hospital general. Material y métodos : Serie de casos comparativa de 193 pacientes con LC tratados en tres ensayos clínicos con AM (n=69) y EGS (n=124) durante 2001-2010. La administración de ambas drogas fue vía endovenosa lenta de 20 mg Sb5+/kg/día por 20 días consecutivos siguiendo las normativas de la OPS y OMS. La información clínica, toxicidad y eficacia fue obtenida de las historias clínicas almacenadas en el centro de investigación según la normativa local e internacional. Resultados : Las características demográficas fueron similares entre grupos, pero el tamaño y número de lesiones fueron mayores en el grupo AM. La eficacia del tratamiento con AM fue 76,0% versus 68,4% con EGS (p=0,340) y 55,1% versus 50,8% (p=0,570) en el análisis por protocolo y de intención de tratar, respectivamente. No se observaron efectos adversos inmediatos. Los síntomas más frecuentemente reportados fueron disgeusia (37,0%), mareos (32,0%), cefalea (36,0%), artralgias (31,0%) y linfangitis (21,0%). Los tres primeros síntomas, así como elevación de transaminasas, leucopenia, trombocitopenia y QTc prolongado fueron frecuentes en el grupo EGS, pero clínica y estadísticamente no significativos. El tratamiento fue suspendido definitivamente por toxicidad severa únicamente con EGS por emesis refractaria (2 participantes) y QTc prolongado con extrasístoles (1 participante). Conclusiones : La eficacia del tratamiento con AM y EGS fue comparable. La administración endovenosa de ambos no produjo efectos adversos inmediatos, aunque sí alteraciones clínicas y laboratoriales usuales.

SUMMARY Objectives : To compare the efficacy and safety of sodium stibogluconate (SS) and meglumine antimoniate (MA) in the treatment of cutaneous leishmaniasis (CL) in a general hospital. Methods: Case-series of 193 patients with CL treated in three clinical trials with MA (n=69) and SS (n=124) during 2001-2010. Both study drugs were administered intravenously at a slow speed at 20 mg Sb5+/kg/day for 20 consecutive days following WHO-PAHO recommendations. Clinical and safety data were gathered from clinical files. Results: Demographic characteristics were similar between the study groups, but the size and number of lesions were higher in the MA group. Efficacy was 76.0% in the MA vs. 68.4% in the SS group (p=0.340) and 55.1% vs. 50.8% (p=0.570) in the per protocol and intention to treat analysis. respectively. Side effects more frequently reported were dysgeusia (37.0%). dizziness (32.0%). headache (36.0%). arthralgia (31.0%) and lymphangitis (21.0%). These first three symptoms as well as elevation of transaminases, leukopenia, thrombocytopenia and prolonged QTc were numerically more frequent in the SS group but without reaching statistical significance. Treatment was stopped definitively for severe toxicity in the SS group due to refractory emesis (two patients) and prolonged QTc (one patient). Conclusions: The efficacy of MA and SS is comparable. The intravenous administration of these compounds did not produce immediate reactions, but it was associated with unusual clinical and laboratory abnormalities.

Uso de anfotericina B liposomal en leishmaniasis cutánea diseminada causada por Leishmania braziliensis en un paciente pediátrico con síndrome de Down / Use of liposomal anfotericin B in diseminated cutaneous leishmaniasis caused by Leishmania braziliensis in a pediatric patient with Down syndrome

Se presenta un caso de leishmaniasis selvática cutánea diseminada con manifestación extensa en una paciente pediátrica con síndrome de Down. El caso se confirmó a través de estudios parasitológicos e inmunológicos, mientras que la identificación se realizó mediante la técnica de reacción en cadena de la polimerasa-polimorfismos de longitud de fragmentos de restricción (PCR-RFLP, por sus siglas en inglés), determinándose la especie como Leishmania (Viannia) braziliensis. La manifestación clínica agresiva y prolongada con poca respuesta a estibogluconato y anfotericina desoxicolato pueden deberse al déficit inmunológico que se presenta como parte del síndrome de Down. La paciente eventualmente recibió tratamiento con anfotericina B liposomal y al término de la terapia, mostró mejoría clínica de las lesiones. El presente reporte ilustra los desafíos tanto de diagnóstico como tratamiento de leishmaniasis cutánea en pacientes pediátricos inmunosuprimidos, especialmente en un entorno de difícil acceso social, económico y geográfico, a los servicios de salud. Se recomienda considerar a la leishmaniasis en el diagnóstico diferencial cuando se atienda ulceras crónicas dermatológicas atípicas; así como tener en cuenta el uso de anfotericina liposomal en pacientes inmunocomprometidos.

We present a case of disseminated cutaneous leishmaniasis with extensive manifestation in a pediatric patient with Down syndrome. The case was confirmed by parasitological and immunological tests. The species was identified as Leishmania (Viannia) braziliensis by polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP). The immune deficit that occurs as part of Down syndrome may have been the reason for the aggressive and prolonged clinical manifestations as well as the poor response to stibogluconate and deoxycholate amphotericin. The patient was treated with liposomal amphotericin B and at the end of therapy, showed clinical improvement of the lesions. This report highlights the challenges of the diagnosis and treatment of cutaneous leishmaniasis in immunosuppressed pediatric patients, especially under difficult social, economic and geographic conditions. Leishmaniasis should be considered as a differential diagnosis when treating atypical chronic dermatologic ulcers; the use of liposomal amphotericin in immunocompromised patients should also be considered in these cases.

Mapeando a leishmaniose visceral e tegumentar americana em Birigui-Sp: uma análise epidemiológica retrospectiva e geoespacial / Mapping american visceral and tegumentary leishmaniasis in Birigui-Sp: a retrospective and geospatial epidemiological analysis / Mapeo de la leishmaniasis visceral y tegumentaria americana en Birigui-Sp: un análisis epidemiológico retrospectivo y geoespacial

A leishmaniose é uma doença tropical infecciosa negligenciada com dois espectros clínicos, a Leishmaniose Visceral (LV) e a Leishmaniose Tegumentar Americana (LTA). O objetivo foi determinar a evolução epidemiológica e a distribuição espacial de LV e LTA em Birigui/SP, entre 2010 a 2020. Foi realizado um estudo retrospectivo e observacional, de LV e LTA registrados pelo Serviço de Vigilância Epidemiológica e os endereços foram utilizados para a análise espacial e distribuição dos casos dentro do município. Identificaram-se 233 casos de LV, com maior número em 2012 (15,45%) e 14 óbitos por LV (letalidade de 6,0%), com predomínio de indivíduos do sexo masculino (60,1%), brancos (71,7%), com idades entre 40 e 59 anos (28,8%), ensino médio completo (8,58%) e alta taxa de cura (65,7%) após tratamento. Verificou- se 25 casos de LTA sendo 2015 o ano com maior número de casos (22,2%) e com predomínio em homens (63%), maiores de 60 anos (44,4%), brancos (92,6%) e com ensino fundamental incompleto (18,5%). A manifestação clínica mais observada foi a forma cutânea (59,3%) com mais de 77% de cura após tratamento. Padrões significativos de agregação espacial foram detectados e áreas de cluster acumulam-se na região oriente do município de Birigui, com maior frequência em bairros mais periféricos. Conclui-se que, apesar de ser observada uma leve diminuição nos casos de LV e LTA, reforça-se a necessidade do monitoramento frequente, principalmente nas regiões com maior número de casos.

Leishmaniasis is a neglected tropical infectious disease with two clinical spectrums, Visceral Leishmaniasis (VL) and American Cutaneous Leishmaniasis (ACL). The objective was to determine the epidemiological evolution and spatial distribution of VL and ATL in Birigui/SP, between 2010 and 2020. spatial analysis and distribution of cases within the municipality. A total of 233 cases of VL were identified, with the highest number in 2012 (15.45%) and 14 deaths from VL (6.0% mortality), with a predominance of males (60.1%), whites (71.7%), aged between 40 and 59 years (28.8%), complete high school (8.58%) and high cure rate (65.7%) after treatment. There were 25 cases of ACL, with 2015 being the year with the highest number of cases (22.2%) and with a predominance of men (63%), over 60 years old (44.4%), white (92.6%) and with incomplete primary education (18.5%). The most observed clinical manifestation was the cutaneous form (59.3%) with more than 77% of cure after treatment. Significant patterns of spatial aggregation were detected, and cluster areas accumulated in the eastern region of the municipality of Birigui, with greater frequency in more peripheral neighborhoods. It is concluded that, despite a slight decrease in cases of VL and ATL, the need for frequent monitoring is reinforced, especially in regions with a higher number of cases.

La leishmaniasis es una enfermedad infecciosa tropical desatendida con dos espectros clínicos, la leishmaniasis visceral (LV) y la leishmaniasis cutánea americana (LCA). El objetivo fue determinar la evolución epidemiológica y distribución espacial de la LV y LCA en Birigui/SP, entre 2010 y 2020. análisis espacial y distribución de los casos dentro del municipio. Fueron identificados 233 casos de LV, con el mayor número en 2012 (15,45%) y 14 muertes por LV (6,0% de mortalidad), con predominio del sexo masculino (60,1%), raza blanca (71,7%), edad entre 40 y 59 años (28,8%), enseñanza media completa (8,58%) y alta tasa de cura (65,7%) después del tratamiento. Hubo 25 casos de LCA, siendo 2015 el año con mayor número de casos (22,2%) y con predominio de hombres (63%), mayores de 60 años (44,4%), blancos (92,6%) y con estudios primarios incompletos (18,5%). La manifestación clínica más observada fue la forma cutánea (59,3%) con más del 77% de curación tras el tratamiento. Se detectaron patrones significativos de agregación espacial, acumulándose áreas de conglomerados en la región este del municipio de Birigui, con mayor frecuencia en barrios más periféricos. Se concluye que, a pesar de una leve disminución de los casos de LV y ATL, se refuerza la necesidad de monitoreo frecuente, especialmente en las regiones con mayor número de casos.

In vitro effects of lapachol and β-lapachone against Leishmania amazonensis

Leishmaniasis is a neglected disease that affects millions of people worldwide, and special attention should be given to treatment because the available drugs have limitations, which can lead to low therapeutic adherence and parasitic resistance. This study evaluated the activity of the bioactive naphthoquinones, lapachol and β-lapachone, against Leishmania amazonensis. The cell alterations were evaluated in vitro on promastigote and amastigote forms. The lethal dose (LD50) at 24, 48, and 72 h on the promastigote's forms using lapachol was 75.60, 72.82, and 58.85 μg/mL and for β-lapachone was 0.65, 1.24, and 0.71 μg/mL, respectively. The naphthoquinones significantly inhibited the survival rate of L. amazonensis amastigotes at 83.11, 57.59, and 34.95% for lapachol (82.28, 41.14, and 20.57 µg/mL), and 78.49, 83.25, and 80.22% for β-lapachone (3.26, 1.63, and 0.815 µg/mL). The compounds on the promastigote's forms led to the loss of mitochondrial membrane potential, induced changes in the integrity of the membrane, caused damage to cells suggestive of the apoptotic process, and showed inhibition of tumor necrosis factor (TNF)-α and interleukin (IL)-6 production. The results showed that these naphthoquinones are promising candidates for research on new drugs with anti-Leishmania activity derived from natural products.

DC-SIGN receptor is expressed by cells from cutaneous leishmaniasis lesions and differentially binds to Leishmania (Viannia) braziliensis and L. (Leishmania) amazonensis promastigotes

BACKGROUND Dendritic cells (DCs) specific intercellular adhesion molecule (ICAM)-3-grabbing non integrin receptor (DC-SIGN) binds to subgenera Leishmania promastigotes mediating its interaction with DC and neutrophils, potentially influencing the infection outcome. OBJECTIVES In this work, we investigated whether DC-SIGN receptor is expressed in cells from cutaneous leishmaniasis (CL) lesions as well as the in vitro binding pattern of Leishmania (Viannia) braziliensis (Lb) and L. (L.) amazonensis (La) promastigotes. METHODS DC-SIGN receptor was labeled by immunohistochemistry in cryopreserved CL tissue fragments. In vitro binding assay with CFSE-labeled Lb or La promastigotes and RAJI-transfecting cells expressing DC-SIGN (DC-SIGNPOS) or mock-transfected (DC-SIGNNEG) were monitored by flow cytometry at 2 h, 24 h and 48 h in co-culture. RESULTS In CL lesion infiltrate, DC-SIGNPOS cells were present in the dermis and near the epidermis. Both Lb and La bind to DC-SIGNPOS cells, while binding to DC-SIGNNEG was low. La showed precocious and higher affinity to DC-SIGNhi population than to DC-SIGNlow, while Lb binding was similar in these populations. CONCLUSION Our results demonstrate that DC-SIGN receptor is present in L. braziliensis CL lesions and interact with Lb promastigotes. Moreover, the differences in the binding pattern to Lb and La suggest DC-SIGN can influence in a difference way the intake of the parasites at the first hours after Leishmania infection. These results raise the hypothesis that DC-SIGN receptor could participate in the immunopathogenesis of American tegumentary leishmaniasis accounting for the differences in the outcome of the Leishmania spp. infection.

Assessing reservoir host status in leishmaniasis with special reference to the infectiousness of Leishmania (Viannia) braziliensis infections in wild rodents

ABSTRACT Molecular methods have been responsible for a notable increase in the detection of Leishmaniinae infections in wild animals. Determining their infectiousness is of paramount importance in evaluating their epidemiological significance. One of the most efficient ways of determining infectiousness for vector borne diseases is xenodiagnosis with the appropriate vector. However, this is logistically very difficult to accomplish in the field, and an ideal solution is to find a molecular surrogate for xenodiagnosis. In this review we discuss different approaches to the problem by focusing on the infectiousness of Leishmania (Viannia) braziliensis in rodents under laboratory and field conditions. Comparisons with similar studies for other Leishmania species emphasizes that there are pivotal differences in the infectiousness and the importance of asymptomatic infections in different hosts. Potentially the most promising surrogate is the real time quantitative PCR (qPCR). However, its success depends on choosing a tissue that relates to the vector's feeding location and the parasite's tissue tropism. This requires detailed knowledge of the infection of each species in its wild hosts. We conclude that for L. (V.) braziliensis infections in wild rodents the tissue of choice for a molecular xenodiagnostic test, based on the qPCR is blood, providing that a significant number of samples must be examined.

Brief communication: Vitamin D serum levels in American tegumentary leishmaniasis from an endemic area in Northeast Brazil

Abstract Introduction The pathogenesis of cutaneous and mucosal leishmaniasis is associated with different immune responses. Vitamin D may modulate the immune system. Here we evaluate the association of vitamin D levels with the severity of the clinical forms of cutaneous and mucosal leishmaniasis. Methods We conducted an observational study evaluating the association between vitamin D levels, disease severity and therapeutic response in patients with cutaneous and mucosal leishmaniasis. Additionally, we conducted a cross-sectional study to compare vitamin D levels in patients with leishmaniasis and healthy subjects. Hypovitaminosis D was defined as a serum level of 25 (OH) D < 30 ng/mL. Results In patients with leishmaniasis, vitamin D serum levels were 38.5 ± 11.54 ng/mL, and 37.5 ± 10.43 ng/mL in healthy subjects The prevalence of hypovitaminosis D was 23.3% and 20.0%, respectively (p = 0.72). There was no correlation between vitamin D serum levels, disease severity, and healing time in the mucosal leishmaniasis group. Conclusion Vitamin D levels are not associated with neither susceptibility nor severity of tegumentary leishmaniasis.

"It's not all about the disease": do treatment and socioeconomic status affect perceived impact and satisfaction of patients treated for cutaneous leishmaniasis?

ABSTRACT Background: This cross-sectional study compared the general impact of cutaneous leishmaniasis (CL) and patient satisfaction with treatment and health services as perceived by those undergoing different therapeutic regimens in an endemic region in South-Eastern Brazil. We also investigated the factors associated with both outcomes (general impact and satisfaction). Methods: We included 84 patients with CL treated between 2018 and 2019 with intravenous meglumine antimoniate, liposomal amphotericin B, or intralesional meglumine antimoniate therapy. Data were collected through interviews that assessed sociodemographic characteristics, comorbidity status, access and use of health services for CL diagnosis and treatment, and the items of the Cutaneous Leishmaniasis Impact Questionnaire (CLIQ). The CLIQ is a psychometric questionnaire previously validated to assess the general impact of CL on patient satisfaction with treatment and health services. Multivariate logistic regression analysis was used to identify the factors associated with high CL impact and low patient satisfaction. Results: The general impact of CL and patient satisfaction with treatment and health services were not significantly associated with the therapeutic regimen. High CL impact was associated with low family income (odds ratio [OR]:3.3; 95% confidence interval [CI]:1.0-10.3), occurrence of complications/adverse effects during treatment (OR:7.7; 95%CI:2.4-25.6), and additional costs during diagnosis and/or treatment (OR:12.1; 95% CI:2.8-52.4). Low satisfaction was associated with high disease impact (OR: 9.5; 95% CI:2.7-33.9), occurrence of complications/adverse effects (OR:4.2; 95% CI:1.3-13.0), and high family income (OR:7.1; 95%CI:1.7-28.2). Conclusions: Our data support public health policies aimed at reducing the impact of CL and its treatment as well as the use of therapy with fewer adverse effects.

Factors associated with mucosal involvement in tegumentary leishmaniasis: a nation-based study using surveillance data from Brazil

ABSTRACT This study aimed to assess the factors associated with mucosal leishmaniasis (ML) within the scope of tegumentary leishmaniasis (TL) cases reported in Brazil. Surveillance data were assessed, and comparisons were made between ML and cutaneous leishmaniasis (CL) cases. Additionally, ML incidence rates for municipalities were depicted through a geographic information system. From 2007 to 2017, 235,489 TL cases were reported, of which 235,232 were classified as follows: 14,204 (6%) were ML cases and 221,028 (94%) were CL cases. Multivariate analysis showed that the proportion of ML cases reached 16.8% among individuals >75 years (adjusted OR = 2.77; 95% CI = 2.41-3.19; p < 0.001), and ML was also more frequent among males (aOR = 1.28; 95% CI = 1.20-1.38; p < 0.001), HIV-positive patients (aOR = 2.15; 95% CI = 1.80-2.56; p < 0.001), patients residing in urban areas (aOR = 1.52; 95% CI = 1.43-1.62; p < 0.001), and imported cases (with respect to county) when compared to autochthonous cases (aOR = 1.84; 95% CI = 1.71-1.98; p < 0.001). A lower proportion of positive results in direct parasitological examinations was observed in ML cases (32.6% vs. 60.8%; p < 0.001). The leishmanin skin test results were more often positive in ML cases (41.7% vs. 25.9%; p < 0.001). In ML, compatible changes in histopathology were more frequent (14.6% vs. 3.9%; p < 0.001). A greater proportion of ML cases were treated with amphotericin B (6.9% vs. 0.9%; p < 0.001). The case-fatality rate was higher in ML (0.6% vs. 0.1%; p < 0.001). A higher incidence of ML was observed in a geographical band extending across the Amazon region from the southern Para State to the Acre State. ML exhibited varying frequencies within specific populations. The definition of predictable factors predisposing Leishmania-infected subjects to develop ML is important for defining strategies to mitigate the mucosal damage caused by leishmaniasis.

Can domestic dogs be considered a good reservoir of Leishmania (L.) infantum chagasi in an endemic area of nonulcerated cutaneous leishmaniasis in Southern Honduras?

ABSTRACT Dogs are considered to be the main domestic reservoir associated with the transmission of Leishmania (L.) infantum chagasi to humans in endemic areas of visceral leishmaniasis in America. However, little is known about the role of canines as a source of infection in endemic areas of nonulcerated cutaneous leishmaniasis (NUCL). Therefore, the objective of the present study was to investigate the role of dogs as a possible reservoir of the parasite in Southern Honduras. Dogs (n = 107) living with individuals affected by NUCL were clinically examined and biological material was collected for parasitological and immunological diagnosis. Most animals showed a healthy appearance and a few presented slight weight loss (64%), alopecia (7%), onychogryphosis (5%) and skin lesions (1%). The overall seroprevalence of Leishmania infection based on the DDP ® quick test and/or in-house ELISA serological test was 41%. The presence of the parasite's DNA was confirmed in 94% of the dogs; however, the average parasite load in the buffy coat was low at 6.09 parasites/µL, ranging between 0.221 and 50.2. The skin of seropositive dogs examined by histopathology using paraffin sections stained by hematoxylin and immunohistochemistry did not show cutaneous lesions or parasite amastigotes. Based on the absence of parasites in the skin and the low parasite load detected in the buffy coat, it seems that the dog does not represent a good source of infection for the vector in the endemic area of NUCL transmission in Southern Honduras. Other domestic and/or wild animals should be investigated.

First report of Leishmania (Mundinia) martiniquensis in South American territory and confirmation of Leishbunyavirus infecting this parasite in a mare

BACKGROUND Epidemiological data related to leishmaniases or Leishmania infection in horses are scarce. However, studies carried out in different regions in the world showed equids parasitised by Leishmania braziliensis, L. infantum and L. martiniquensis. OBJECTIVES Identify the Leishmania species causing cutaneous leishmaniasis in a mare, living in Rio de Janeiro State (Brazil), and search the presence of Leishmania viruses in the isolated parasite. METHODS Isoenzymes and polymerase chain reaction (PCR) targeting ITSrDNA region followed by sequencing were conducted for typing the isolated parasite. A search for Leishmania virus infection was also performed. FINDINGS The mare presented skin nodules and ulcers in the left pinna caused by Leishmania spp. that was detected by culture and PCR. The parasite was identified as Leishmania (Mundinia) martiniquensis, infected by Leishbunyavirus (LBV), representing the first description of this species in South America. The animal travelled to different Brazilian regions, but not to outside the country. MAIN CONCLUSIONS The worldwide distribution of L. martiniquensis and its infection by LBV were confirmed in this study, indicating the autochthonous transmission cycle in Brazil. The clinical profile of the disease in the mare, showing fast spontaneous healing of cutaneous lesions, may indicate that skin lesions related to L. martiniquensis infection in horses might be underdiagnosed.

Prácticas y saberes de excombatientes de las FARC-EP sobre el tratamiento de la leishmaniasis cutánea / Practices and knowledge of ex-combatents of the FARC-EP on the treatment of cutaneous leishmaniasis

RESUMEN Introducción: La Leishmaniasis es una enfermedad infecciosa parasitaria zoonótica que afecta la piel, las mucosas y las vísceras. Es endémica en 98 países y hay más de 350 millones de personas en riesgo de transmisión. En Colombia, los campamentos de las antiguas Fuerzas Armadas Revolucionarias de Colombia, se ubicaban en zonas selváticas donde la enfermedad está presente. El objetivo del presente trabajo está dirigido a describir las prácticas y saberes de los excombatientes de las FARC-EP sobre la Leishmaniasis cutánea y su manejo en escenarios de combate al convivir con esta enfermedad. Métodos: El estudio es cualitativo/exploratorio, donde se abordaron los conocimientos sobre la enfermedad y su forma de transmisión, como también aquellas prácticas relacionadas con el uso de medicamentos para su tratamiento. Resultados: En el análisis de las entrevistas se encontró que los excombatientes conocen el ciclo de la enfermedad desde sus vivencias y por transmisión oral, como también de prácticas curativas propias adaptadas al contexto del conflicto armado, esta experiencia los llevó a desarrollar estrategias de gestión del conocimiento basado en la evidencia. Discusión: Las entrevistas dan cuenta del conocimiento empírico y clínico de los excombatientes sobre el ciclo de la enfermedad; no existe un registro documental, ni manuales de procedimientos realizados en la selva por las FARC-EP sobre tratamiento de enfermedades; los miembros de las FARC-EP a desarrollar estrategias de gestión del conocimiento basados en la evidencia; el contexto de conflicto afectó de manera importante el tiempo y los recursos técnicos para manejar la Leishmaniasis, esto posibilitó las respuestas alternativas a la enfermedad documentadas en este trabajo.

ABSTRACT Introduction: Leishmaniasis is a zoonotic parasitic infectious disease that affects the skin, mucous membranes and viscera. It is endemic in 98 countries and there are more than 350 million people at risk of transmission. In Colombia, the camps of the former Revolutionary Armed Forces of Colombia were located in jungle areas where the disease is present. The objective of this work is aimed at describing the practices and knowledge of ex-combatants of the FARC-EP about cutaneous Leishmaniasis and its management in combat scenarios when living with this disease. Methods: The study is qualitative/exploratory, where knowledge about the disease and its form of transmission were addressed, as well as those practices related to the use of medications for its treatment. Results: In the analysis of the interviews it was found that the ex-combatants know the cycle of the disease from their experiences and by oral transmission, as well as their own healing practices adapted to the context of the armed conflict, this experience led them to develop knowledge management strategies based on the evidence. Discussion: The interviews give an account of the empirical and clinical knowledge of the ex-combatants about the cycle of the disease; There is no documentary record, nor manuals of procedures carried out in the jungle by the FARC-EP on the treatment of diseases; the members of the FARC-EP to develop evidence-based knowledge management strategies; the context of conflict significantly affected the time and technical resources to manage Leishmaniasis, this made possible the alternative responses to the disease documented in this work.

American Cutaneous Leishmaniasis in Eastern Minas Gerais: veiled reality of a neglected disease / Leishmaniose Tegumentar Americana no Leste de Minas Gerais: realidade velada de uma doença negligenciada

Objective: to analyze the notified and confirmed cases of ACL in a municipality in east Minas Gerais, from 2007 to 2020. Methods: a combined study was carried out as a cross-sectional and an ecological approach of time series type, using notified and confirmed ACL cases, from 2007 to 2020. Primary and secondary data were used. Data were analyzed using descriptive and inferential statistics (simple linear regression, T-test, Mann-Whitney, chi-square (χ2) at a 5% significance level). Results: a total of 219 cases were reported with a decreasing temporal trend, with a higher frequency observed for the cutaneous form (82.6%), age group 40 to 59 years (32.1%), black race (56.4%), and completed elementary school (47.7%). Individuals with the mucosal clinical form had lesions for a longer time, a greater chance of not progressing to cure, and used more vials of meglumine antimoniate when compared to patients with the cutaneous form. Conclusions: different correlations were observed between the variables studied and the profile of involvement described in the scientific literature, with the clinical form predominantly cutaneous and with a good prognosis.

Objetivo: analisar os casos notificados e confirmados de LTA em um município do leste de Minas Gerais, no período de 2007 a 2020. Métodos: foi realizado um estudo combinado com abordagem transversal e ecológica do tipo série temporal, utilizando casos notificados e confirmados de LTA, de 2007 a 2020. Foram utilizados dados primários e secundários. Os dados foram analisados por meio de estatística descritiva e inferencial (regressão linear simples, teste T, Mann-Whitney, qui-quadrado (χ2) com nível de significância de 5%). Resultados: foram notificados 219 casos com tendência temporal decrescente, com maior frequência observada para a forma cutânea (82,6%), faixa etária de 40 a 59 anos (32,1%), raça negra (56,4%) e ensino fundamental completo (47,7%). Indivíduos com a forma clínica mucosa apresentaram maior tempo de lesão, maior possibilidade de não evoluir para cura e utilizaram mais ampolas de antimoniato de meglumina quando comparados aos pacientes com a forma cutânea. Conclusões: foram observadas diferentes correlações entre as variáveis estudadas e o perfil de acometimento descrito na literatura científica, com a forma clínica predominantemente cutânea e com bom prognóstico.

Análisis del comportamiento epidemiológico de Leishmaniasis cutánea en Perú 2016-2021 / Analysis of the epidemiological behavior of cutaneous leishmaniasis in Peru 2016-2021

La leishmaniasis cutánea (LC) es una patología cutánea inmunomediada causada principalmente por Leishmania (L.) major, Leishmania tropica, Leishmania braziliensis, L. mexicana y L. amazonensis. La carga de la LC en términos de morbilidad y estigmas sociales se concentra en ciertos países en desarrollo de Asia, África y Sudamérica. Las personas con LC asintomática representan una gran proporción de los individuos infectados en las zonas endémicas que no presentan ninguna lesión y pueden controlar la infección mediante mecanismos aún no del todo conocidos. En la actualidad, no existe ninguna medida profiláctica de control aprobada para la LC. Este reporte epidemiológico reveló que la LC prevalecía en las áreas geográficas de Selva y Sierra de Perú, mientras que departamentos como Ancash y La Libertad, áreas de la Costa, también estuvieron involucradas en el reporte de casos en los últimos años. El patrón de casos de LC reportados en los distintos departamentos del país tuvo un comportamiento muy parecido en el periodo comprendido entre 2016 y 2021, predominando en las primeras 4 o 5 semanas epidemiológicas la mayor frecuencia de pacientes enfermos con lesiones dérmicas compatibles y corroboradas con LC. La enfermedad es endémica en muchas partes del país, por eso se necesitan más esfuerzos y enfoques integrados para combatir esta patología(AU)

Cutaneous leishmaniasis (CL) is an immune-mediated skin pathology caused mainly by Leishmania (L.) major, Leishmania tropica, Leishmania braziliensis, L. mexicana and L. amazonensis. The burden of CL in terms of disease and social stigma is concentrated in certain developing countries in Asia, Africa, and South America. People with asymptomatic CL represent a large proportion of infected individuals in endemic areas who do not have any lesions and can control the infection through mechanisms that are not fully understood. Currently, there is no approved prophylactic control measure for CL. This epidemiological report revealed that CL was prevalent in the Selva and Sierra geographic areas of Peru, while departments such as Ancash and La Libertad, coastal areas, were also involved in the reporting of cases in recent years. The pattern of CL cases reported in the different departments of the country had a very similar behavior in the period between 2016 and 2021, with the highest frequency of sick patients with skin lesions compatible and corroborated with CL predominating in the first 4 or 5 epidemiological weeks. The disease is endemic in many parts of the country, so more efforts and integrated approaches are needed to combat this pathology(AU)

Eficacia de la artemisina en modelos murinos de leishmaniasis cutánea experimental causada por Leishmania amazonensis / Artemisine efficacy in murine models of experimental cutaneous leishmaniasis caused by Leishmania amazonensis

Los limitados tratamientos disponibles para enfrentar la leishmaniasis requieren el desarrollo de investigaciones para buscar nuevos agentes terapéuticos. Una estrategia recomendada es el reposicionamiento farmacológico, en el que la artemisina figura como un posible candidato. El objetivo de este estudio es evaluar las potencialidades de la artemisina en dos modelos murinos de leishmaniasis cutánea experimental. Para ello, se emplearon ratones BALB/c (susceptibles) y C57BL/6 (resistentes) infectados con Leishmania amazonensis. El tratamiento se realizó por vía oral o intralesional con cinco dosis de artemisina a 30 mg/kg cada 4 días. Se determinó el comportamiento del peso, la evolución del tamaño de la lesión y la carga parasitaria. En ambos modelos animales se observó que el tratamiento con artemisina (oral e intralesional) disminuyó el tamaño de la lesión y la carga parasitaria con respecto a los grupos infectados sin tratamiento (p 0,05). Los ratones C57BL/6 tratados por vía oral fueron los únicos capaces de controlar las lesiones hasta el final del experimento. Se demuestra la eficacia in vivo de la artemisina en dos modelos de leishmaniasis cutánea inducida por L. amazonensis y se destaca la administración por vía oral en el control de la enfermedad. Se sugiere el futuro desarrollo de este fármaco para el tratamiento de la leishmaniasis cutánea.

The limited treatments available for leishmaniasis require the development of research for new therapeutic agents. One recommended strategy is the pharmacological repositioning, where artemisinin stands out as a possible candidate. The aim of this study is to evaluate the potential of artemisinin in two murine models of experimental cutaneous leishmaniasis. For this purpose, BALB/c (susceptible) and C57BL/6 (resistant) mice infected with Leishmania amazonensis were used. Oral or intralesional treatment was performed with five doses of artemisinin at 30 mg/kg every four days. Weight behavior, evolution of lesion size, and parasitic load were determined. In both animal models it was observed that treatment with artemisinin (oral and intralesional) decreased lesion size and parasitic load with respect to the untreated infected groups (p 0.05). Orally treated C57BL/6 mice were the only ones able to control lesions until the end of the experiment. The in vivo efficacy of artemisinin in two models of cutaneous leishmaniasis induced by L. amazonensis is demonstrated and oral administration is highlighted in the control of the disease. Further development of this drug for the treatment of cutaneous leishmaniasis is suggested.

Bases para un tratamiento con hipertermia moderada en leishmaniosis cutánea / Bases for a treatment of cutaneous leishmaniasis with moderate hyperthermia

Resumen Introducción: La hipertermia regional entre 38 y 39.5 °C ha sido empleada para tratar procesos inflamatorios y, ocasionalmente, infecciones cutáneas. En zonas endémicas de leishmaniosis se aplican compresas calientes como tratamiento antiparasitario. Objetivo: Conocer las bases del tratamiento térmico de la leishmaniosis para regularlo adecuadamente. Métodos: Parásitos Leishmania mexicana cultivados in vitro fueron incubados por periodos variables de 37 °C y después a 25 °C.. Los parásitos se tiñeron con anexina V-FITC y yoduro de propidio para detectar inducción de apoptosis y su viabilidad. Se realizaron curvas de crecimiento postratamiento e identificación del ciclo celular con anticuerpos anticiclinas. Resultados: Después de 30 minutos de exposición a una temperatura de 37 °C, un porcentaje variable de parásitos perdieron su característica forma ovalada y se tornaron esféricos, sin refringencia y con núcleos condensados, cambios que sugirieron apoptosis, la cual fue confirmada mediante tinción con anexina V-FITC. La cantidad de parásitos en proceso de apoptosis fue proporcional al tiempo de exposición. Los parásitos en los que se observó apoptosis se tiñeron con anticuerpos anticiclinas. Conclusiones: La elevación constante, regulada y fisiológica de la temperatura por más de 30 minutos induce apoptosis de parásitos Leishmania mexicana cultivados in vitro, cuando se encuentran en fase activa en el ciclo celular.

Abstract Introduction: Regional hyperthermia at between 38 and 39.5 °C has been used to treat inflammatory processes and, occasionally, skin infections. In areas where leishmaniasis is endemic, hot compresses are applied as anti-parasitic treatment. Objective: To identify the bases of leishmaniasis thermal treatment in order to properly regulate it. Methods: In vitro-cultured Leishmania mexicana parasites were incubated for variable periods at 37 and then, to 25 °C. The parasites were then stained with Annexin V-FITC to detect apoptosis induction and with propidium iodide for viability. Post-treatment growth curves and cell cycle identification with anti-cyclin antibodies were performed. Results: After 30 minutes of exposure to a temperature of 37 °C, a variable proportion of parasites lost their characteristic oval shape and became spherical, without refringence and with condensed nuclei, with these changes suggesting apoptosis, which was confirmed by Annexin V-FITC staining. The number of parasites that underwent apoptosis was proportional to exposure time. Parasites in which apoptosis was observed were stained with anti-cyclin antibodies. Conclusions: Constant, regulated and physiological elevation of temperature for more than 30 minutes induces apoptosis of in vitro-cultured L. mexicana parasites when they are in an active phase of the cell cycle.

New developments in the treatment of cutaneous leishmaniasis

Leishmaniasis, including the cutaneous form, poses an important public health threat around the world, while no vaccine is currently available against any form of leishmaniasis. The drugs used in the first line treatment of cutaneous leishmaniasis (CL) are commonly pentavalent antimonials despite their toxicities, long-term treatment duration and increasing resistance rates. Other alternatives are amphotericin B, pentamidine, miltefosine and paromomycine. Movement of the population, especially in endemic regions, increases the spread of the parasite and affectes the distribution of causative species, which requires re-evaluation the treatment regimen. Extensive researches are carried out on the treatment of leishmaniasis. The immunotherapeutic and targeted therapeutic approaches, formulations of carrier-loaded active drugs, local thermotherapeutic applications, the combination of antileishmanial drugs/compounds, the use of new synthetic and natural products are promising therapeutic options in the future. Herein, the author reviews the potential treatment modalities of CL with a brief overview of current treatments in the light of ongoing studies around the world.