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Background: This study aimed to describe the kinetics of Leishmania parasite load determined using kinetoplast DNA (kDNA)-based quantitative polymerase chain reaction (qPCR) in visceral leishmaniasis (VL) patients., Methods: Parasite load in blood was assessed by qPCR at five time points, up to 12 months post-diagnosis. Sixteen patients were followed up., Results: A significant reduction in the parasite load was observed after treatment (P < 0.0001). One patient had an increased parasite load 3 months post-treatment and relapsed clinically at month six., Conclusions: We have described the use of kDNA-based qPCR in the post-treatment follow-up of VL cases.

The aim of this study was to investigate the level of exposure to Leishmania infection in stray dogs in an area of intense visceral leishmaniasis transmission in the state of Pernambuco, Brazil. Blood samples from 178 dogs were analyzed using serological and molecular assays: rapid immunochromatographic test (ICT), enzyme-linked immunosorbent assay (ELISA), immunofluorescence antibody test (IFAT), and conventional and quantitative polymerase chain reaction (cPCR and qPCR). Positivity values obtained with serological tests were 71.4% (127/178), 70.2% (125/178), and 50.6% (90/178) using ICT, ELISA, and IFAT, respectively, with 38.8% (69/178) of the dogs were simultaneously positive for all three tests. The positivity values obtained with cPCR and qPCR were 20.2% (36/178) and 38.8% (69/178), respectively, with 11.8% (21/178) testing positive in both molecular assays. Overall, 87.1% (155/178) were positive for anti-Leishmania spp. antibodies and/or Leishmania spp. DNA. Positivity to one or more tests was statistically associated with lymphadenomegaly, skin lesions, lymphocytosis, anemia and hyperproteinemia. The results of this study revealed a high level of exposure to Leishmania in stray dogs in an area of intense human visceral leishmaniasis transmission, suggesting that dogs play a role as reservoirs in the transmission cycle of this zoonosis.

Background: The blood-feeding behaviour of female sand flies may increase their likelihood of acquiring and transmitting Leishmania parasites. Studies on the host usage by these insects may thus improve our understanding of the Leishmania transmission risk in leishmaniasis-endemic areas. Here, we developed a fast multiplex real-time PCR assay for simultaneous detection of dog, human and Leishmania DNA in sand flies., Methods: Primers and TaqMan probes targeting the mitochondrial cytochrome c oxidase subunit 1 and cytochrome b genes of dog and human, respectively, were combined in a multiplex assay, which also includes primers and a TaqMan probe targeting the Leishmania minicircle kinetoplast DNA., Results: The multiplex assay was 100% specific, with analytical sensitivities of 10 3 fg/reaction for dog and human and 1 fg for Leishmania. By testing field-collected engorged female sand flies (95 Migonemyia migonei and two Nyssomyia intermedia), 50 M. migonei were positive for one or two targets (positivity rates: 45.4% for human, 4.1% for dog and 12.4% for Leishmania DNA)., Conclusions: This multiplex real-time PCR assay represents a novel fast assay for detecting dog, human and Leishmania DNA in female sand flies and therefore a tool for assessing the risk of Leishmania transmission to these hosts in areas of active transmission.

Cutaneous leishmaniasis (CL) by Leishmania braziliensis Vianna (Kinetoplastida: Trypanosomatidae) is a widespread disease in the western hemisphere and Brazil is the country reporting the majority of the human cases each year. Several phlebotomine sand flies have been regarded as putative or proven vectors of L. braziliensis and multiple vectors may be involved in the transmission of this parasite in the same endemic foci. In this study, we tested a subset of female phlebotomine sand flies collected in the frame of a previous study conducted in a military training camp, where outbreaks of CL by L. braziliensis have sporadically been reported. In total, 1,807 female phlebotomine sand flies were tested (individually or in pools) by real-time PCR and a minimum infection rate of 0.9% (95% CI: 0.6-1.5%) was detected. Positive females belonged to the species Psychodopygus complexus (Mangabeira) or Psychodopygus wellcomei Fraiha, Shaw & Lainson (females of these species are morphologically indistinguishable), Trichopygomyia longispina (Mangabeira), and Sciopemyia sordellii (Shannon & Del Ponte) (Diptera: Psychodidae). By restriction enzyme analysis, 13 samples (nine of Ty. longispina, two of Ps. complexus/wellcomei, and two of Sc. sordellii) presented a HaeIII restriction profile identical to L. braziliensis. The results of this study reinforce the notion that multiple vectors may be involved in the transmission of L. braziliensis in a single focus, ultimately making the epidemiological picture more complex than currently recognized., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)