Identification of blood meal sources of Lutzomyia longipalpis using polymerase chain reaction-restriction fragment length polymorphism analysis of the cytochrome B gene

Abstract

An analysis of the dietary content of haematophagous insects can provide important information about the transmission networks of certain zoonoses. The present study evaluated the potential of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the mitochondrial cytochrome B (cytb) gene to differentiate between vertebrate species that were identified as possible sources of sandfly meals. The complete cytb gene sequences of 11 vertebrate species available in the National Center for Biotechnology Information database were digested with Aci I, Alu I, Hae III and Rsa I restriction enzymes in silico using Restriction Mapper software. The cytb gene fragment (358 bp) was amplified from tissue samples of vertebrate species and the dietary contents of sandflies and digested with restriction enzymes. Vertebrate species presented a restriction fragment profile that differed from that of other species, with the exception of Canis familiaris and Cerdocyon thous. The 358 bp fragment was identified in 76 sandflies. Of these, 10 were evaluated using the restriction enzymes and the food sources were predicted for four: Homo sapiens (1), Bos taurus (1) and Equus caballus (2). Thus, the PCR-RFLP technique could be a potential method for identifying the food sources of arthropods. However, some points must be clarified regarding the applicability of the method, such as the extent of DNA degradation through intestinal digestion, the potential for multiple sources of blood meals and the need for greater knowledge regarding intraspecific variations in mtDNA.

Identification of blood meals in field captured sand flies by a PCR-RFLP approach based on cytochrome b gene

“…Amplification by polymerase chain reaction (PCR) and subsequent sequencingis widely used to study bloodmeal preferences in sand flies and other vectors as mosquitoes or ticks (Kirstein and Grey 1996, Oshagi et al 2006, Danabalan et al 2014.Thus, different molecular targets like cytochrome b (cytb), cytochrome oxidase I (COI) and prepronociceptin (PNOC) are being used to detect the origin of blood meals (Kocher et al 1989, Haoaus et al 2007, Abassi et al 2009). On the other hand, other techniques as PCR-restriction fragment length polymorphism (RFLP)or PCRreverse-line blotting (RLB) have been developed in order to study blood meal preferences in mosquitoes, tsetse flies and sand flies (Osaghi et al 2006, Steuber et al 2005, Maleki-Ravasan et al 2009, Quaresma et al 2012, Soares et al 2014, Gebresilassie et al 2015.…”

Contribution to the Knowledgment of Feeding Habbits of Lutzomyia (Lutzomyia) Longipalpis (Lutz & Neiva, 1912) in Association to Natural Infection by Leishmania (Leishmania) Infantum Chagasi (Cunha & Chagas, 1937)

“…Several methods are used to identify eating habits in phlebotomines, such as immunological methods, through the precipitin technique [4,8,11,[28][29][30] and immunoenzymatic tests [7,9,15,[31][32]; and recently molecular methods using PCR [33][34][35][36][37][38][39][40][41][42]. Studies that seek to determine the dietary pattern of phlebotomines are of great ecological and epidemiological relevance, since they may at the same time increase the knowledge about the habits of this vector, but mainly suggest potential reservoirs of Leishmania spp., which can help in the planning of better strategies [11,33,37,40].…”

Identification of abnormal nuclear and mitochondrial genes in esophageal cancer cells

“…Furthermore, mtDNA has hologynic features, in which mutations accumulate continuously along the matriarchal line, and the accumulation of mutations increases the risk of tumorigenesis ( 9 ). In addition, mtDNA has more polymorphisms ( 15 ), appearing as significant sequence differences among different races and in different regions. The function of mtDNA is known to be involved in tumorigenesis in head and neck cancer ( 16 ), bladder cancer ( 10 ), breast cancer ( 17 ) and lung cancer ( 11 ).…”