Molecular screening of Leishmania spp. infection and bloodmeals in sandflies from a leishmaniasis focus in southwestern Turkey

Abstract

Leishmaniasis is an arthropod-borne disease that affects approximately 2 million people worldwide annually. The aims of this study were to detect the presence of Leishmania (Kinetoplastida: Trypanosomatidae) DNA and the feeding preferences of probable vector species in an endemic focus of Leishmania infantum in Turkey. Entomological sampling was performed in August and October 2015 in Aydın province, where cases of human and canine leishmaniasis have been reported previously. A total of 1059 sandfly specimens comprising nine species belonging to two genera, Phlebotomus and Sergentomyia (both: Diptera: Psychodidae), and five subgenera of the Phlebotomus genus (Phlebotomus, Paraphlebotomus, Larroussius, Adlerius and Transphlebotomus) were collected in five villages. Among all Phlebotomus specimens, Phlebotomus neglectus (39%) was noted as the most abundant species, followed by Phlebotomus tobbi (18%). Leishmania DNA was detected in pools from P. neglectus, P. tobbi and Sergentomyia dentata by kDNA polymerase chain reaction (PCR). Leishmania DNA from Phlebotomus specimens was identified as L. infantum, but Leishmania DNA from Sergentomyia spp. could not be identified to species level by ITS-1 real-time PCR. The detection of Leishmania DNA in wild-caught P. neglectus and the high percentage (24.2%) of human DNA in engorged specimens suggests that P. neglectus is probably an important vector species for L. infantum in Aydın province.

Real-time PCR applications for diagnosis of leishmaniasis

“…Using qPCR-based approaches, Leishmania DNA has also been detected in sand flies from Albania [ 89 ], Tunisia [ 90 ] and Turkey [ 91 ]. Recently, an interesting approach based on sequential qPCR targeting kDNA followed by ITS1 qPCR was proposed by Karaku et al [ 92 ] to study a leishmaniasis focus in southwestern Turkey. In fact, in this region different Leishmania species coexist, therefore a genus-specific kDNA SYBR-Green qPCR was used to screen for infection in the collected sandflies and qPCR targeting ITS1 region was performed using species-specific primers for detecting L. donovani/infantum complex, L. tropica and L. major .…”

“…In fact, in this region different Leishmania species coexist, therefore a genus-specific kDNA SYBR-Green qPCR was used to screen for infection in the collected sandflies and qPCR targeting ITS1 region was performed using species-specific primers for detecting L. donovani/infantum complex, L. tropica and L. major . Moreover, the DNA extracted from the abdomens of freshly blood-fed female sandflies was amplified using the cytochrome b gene as a target to identify the vertebrate source of the blood meal and to reveal the host preferences in the study area [ 92 ]. Interestingly, the qPCR approach was utilized also to detect L. infantum DNA in ticks ( Rhipicephalus sanguineus ) removed from dogs living in endemic areas in Italy, finding the parasite DNA in a fraction of ticks examined [ 93 , 94 ].…”

Leishmania sp. detection and blood‐feeding behaviour of S ergentomyia minuta collected in the human leishmaniasis focus of southwestern Madrid, Spain (2012–2017)

“…Sand flies from this genus are traditionally involved in the transmission of Sauroleishmania species and are known for their preference to feed on cold‐blooded animals. However, recent studies have detected DNA from other vertebrates, including humans, in blood‐fed females of different species of Sergentomyia (Azizi, Askari, Kalantari, & Moemenbellah‐Fard, ; Berdjane‐Brouk et al, ; Bravo‐Barriga et al, ; Karakuş et al, ; Maia et al, ; Senghor et al, ; Siripattanapipong, Leelayoova, Ninsaeng, & Mungthin, ). Also, several entomological studies carried out in Old World countries have shown the presence of DNA from pathogenic Leishmania species in Sergentomyia genus.…”

Effect of trapping method on species identification of phlebotomine sandflies by MALDI‐TOF MS protein profiling

“…Although they facilitate a partial catch of live specimens, one of the limitations of CDC light traps is their presumably selective attraction of different species and sexes (Alexander, ; Alten et al ., ). Therefore, sticky traps have continued to be used in recent studies that aim to investigate sandfly fauna composition (Sawalha et al ., ) and resting and breeding sites (Khan et al ., ), or to detect the presence of Leishmania in captured females (Karakus et al ., ).…”

“…Although they facilitate a partial catch of live specimens, one of the limitations of CDC light traps is their presumably selective attraction of different species and sexes (Alexander, 2000;Alten et al, 2015). Therefore, sticky traps have continued to be used in recent studies that aim to investigate sandfly fauna composition (Sawalha et al, 2017) and resting and breeding sites (Khan et al, 2017), or to detect the presence of Leishmania in captured females (Karakus et al, 2017). The technique of MALDI-TOF MS protein profiling represents an efficient approach towards species identification as this method has important benefits, including simple sample preparation, and extremely fast data acquisition and evaluation, and requires only inexpensive consumables.…”