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Brazilian Journal of Biology

Print version ISSN 1519-6984On-line version ISSN 1678-4375

Braz. J. Biol., ahead of print  Epub Feb 17, 2020

http://dx.doi.org/10.1590/1519-6984.228178 

Notes and Comments

Identification of sand flies (Diptera: Psychodidae) and blood meal sources in periurban areas of Ji-Paraná municipality, Western Brazilian Amazon

aPrograma de Pós-graduação em Biologia Experimental, Fundação Universidade Federal de Rondônia, BR 364, Km 9.5, CEP 76801-059, Porto Velho, RO, Brasil

bLaboratório de Entomologia, Fundação Oswaldo Cruz – Fiocruz Rondônia, Rua da Beira, 7671, CEP 76812-245, Porto Velho, RO, Brasil

cInstituto Nacional de Epidemiologia da Amazônia Ocidental – INCT-EpiAMO, Rua da Beira, 7671, CEP 76812-245, Porto Velho, RO, Brasil

dLaboratório de Epidemiologia Genética, Fundação Oswaldo Cruz – Fiocruz Rondônia, Rua da Beira, 7671, CEP 76812-245, Porto Velho, RO, Brasil


Leishmaniases is a disease caused by Leishmania protozoans which are transmitted to vertebrates by the bite of female sand flies. Epidemiological surveillance depends on the identification of vector species and their reservoirs, and identifying sand fly blood meal sources can be an effective means of identifying potential reservoirs in the Leishmaniasis transmission cycle (Roque and Jansen, 2014). In Rondônia State, approximately 1,000 new cases of cutaneous leishmaniasis (CL) occur annually (DATASUS, 2018). In Ji-Paraná municipality, which is located in the eastern mesoregion of Rondônia (see Figure 1), 418 cases of CL have been registered since 2007 (DATASUS, 2018); in spite of this, there is not much information about Leishmania infection, sand fly vectors and, their blood sources that occur there (Biancardi et al., 1982). The aims of our study was to verify these insects and possible blood meals in Ji-Paraná municipality.

Figure 1 (A) South America map with Brazil Map delimited by white line indicating the states limits and in yellow the limits of the Rondônia State; (B) Map of Rondônia state in a large view (yellow line) and in green line indicating the limits of Ji-Paraná municipality; (C) Collection points distributed among periurban areas of Ji-Paraná with the number of Psychodopygus davisi (blue) and Ps. hirsutus hirsutus individuals (red) given for each location. 

Sand fly collections were made using light traps that were set for three consecutive nights in small forest fragments near eight periurban areas (see Figure 1C). Two collections were performed: one in November of 2017 and another in November of 2018. Males and females (head and last segments) were slide-mounted and morphologically identified (Galati, 2018).

In order to screen for the presence of Leishmania DNA, the thoraxes of non-engorged females were separated into pools and subjected to polymerase chain reaction assays using primers targeting kDNA (Pereira Júnior et al., 2019). The Le. amazonensis strain (IFLA/BR/1967/PH8) was used as the positive control and ultrapure water was used as the negative control. Blood meal sources were identified by submitting the thoraxes of engorged females to DNA extraction using the phenol/chloroform protocol; to avoid sample contamination, two samples containing a female with no blood present in the gut and one sample containing DNA-free water were used as negative controls. Extracted DNA was subjected to PCR assays targeting the cytb mitochondrial region (Pereira Júnior et al., 2019).

A total of 1,331 sand flies comprising 28 species were identified (as shown in Table 1). The most abundant species were Psychodopygus davisi (491 individuals, 36.9%) and Ps. hirsutus hirsutus (391 individuals, 29.4%) (see Figure 1C). In Rondônia, Ps. davisi generally occurs in high abundance (Gil et al., 2003; Pereira Júnior et al., 2019). Specimens of both species have been found carrying Leishmania naiffi promastigotes (Gil et al., 2003); however, none of the 80 pools in our study exhibited the presence of Leishmania DNA.

Table 1 Sand fly fauna collected at eight points in Ji-Paraná municipality in November of 2017 and 2018. Numbers in parentheses represent engorged females. 

Species P1 P2 P3 P4 P5 P6 P7 P8 Total %
♀/♂ ♀/♂ ♀/♂ ♀/♂ ♀/♂ ♀/♂ ♀/♂ ♀/♂
Bichromomyia flaviscutellata 5/2 1/0 1/5 - 0/2 0/2 2/4 - 24 1.8
Brumptomyia brumpti - - 4/7 1/3 - - - - 15 1.1
Evandromyia saulensis 2/0 - 45/28 5/1 1/0 2/0 5/0 - 89 6.7
Evandromyia walkeri 2/1 1/0 8/7 - - - - - 19 1.4
Lutzomyia sherlocki - 1/0 1/0 - - 3/0 - - 5 0.4
Microppygomyia rorotaensis - - - - 1/0 0/2 2/2 - 7 0.5
Micropygomyia villelai 0/3 - - - - 2/0 2/0 - 7 0.5
Migonemyia migonei - 1/0 1/0 - - - - - 2 0.2
Nyssomyia antunesi 8/0 1/3 24/5 2/0 4/0 7/0 12/3 - 69 5.2
Nyssomyia whitmani 1/0 0/1 2/2 - 1/2 4/1 10/3 0/1 28 2.1
Pintomyia nevesi 5/6 2/5 2/1 - - 2/0 - 2/3 28 2.1
Pintomyia serrana 0/3 - 1/0 - - 1/0 - - 5 0.4
Psathyromyia campelli - - - - - - 1/0 - 1 0.1
Psathyromyia dendrophyla - - 2/0 - - - - - 2 0.2
Psathyromyia hermanlenti - - - 0/1 1/0 0/3 - - 5 0.4
Psathyromyia lutziana 1/0 - 1/0 - - - 0/1 - 3 0.2
Psathyromyia runoides 0/1 - 0/1 - - - - - 2 0.2
Psychodopygus ayrozai - - 1/0 - - - - - 1 0.1
Psychodopygus c. carrerai - 4/4 5/1 2/1 5/0 9/18 5/3 - 57 4.3
Psychodopygus chagasi - 1/0 3/0 3/0 1/0 - 8/0 - 16 1.2
Psychodopygus claustrei 0/4 0/1 1/6 0/1 0/4 0/3 - 0/1 21 1.6
Psychodopygus complexus - - 0/1 - 0/2 - 1/16 - 20 1.5
Psychodopygus davisi 25/82 14/26 38/85 4/3 94(1)/53 19/28 4/10 4/2 491 36.9
Psychodopygus geniculatus - - - - 0/1 2/3 - - 6 0.5
Psychodopygus h. hirsutus 6/10 2/2 49/35 8(1)/14 103/22 65(3)/52 15/6 2/0 391 29.4
Sciopemyia servulolimai - - - - - - 1/1 - 2 0.2
Sciopemyia sordellii - - - - - 0/1 8/2 - 11 0.8
Trichophoromyia clitella - 0/1 0/2 - - 0/1 - - 4 0.3
Total 167 71 375 49 297 230 127 15 1,331 100

Blood meal analysis revealed the DNA of Homo sapiens in one Ps. davisi female and four Ps. hirsutus hirsutus females, while DNA of the wild vertebrates Alouatta seniculus and Dasypus sabanicola was present in the gut of two Ps. hirsutus hirsutus females (as shown in Table 2). Dasypus sabanicola is not found in Brazil (Abba and Superina, 2010) and probably the DNA sequence could be another species of Dasypus genera with distribution in the Brazilian Amazon as Dasypus novemcinctus or Dasypus kappleri.

Table 2 Vertebrate species identified from sand fly females collected in Ji-Paraná municipality. 

Sand fly species Blood
Meal
Accession Location Identity (%) Total score Query cover (%) E-value
Ps. hirsutus Dasypus sabanicola KT818545.1 Point 3 95.74 448 79 1E-121
Ps. hirsutus Homo sapiens KX697544.1 Point 3 99.01 538 90 5E-149
Ps. hirsutus Homo sapiens LCO88150.1 Point 4 98.27 507 89 2E-139
Ps. davisi Homo sapiens KX697544.1 Point 5 96.58 477 85 1E+130
Ps. hirsutus Homo sapiens LCO88148.1 Point 6 97.82 473 84 2E-129
Ps. hirsutus Homo sapiens MH981639.1 Point 6 98.36 532 86 5E-147
Ps. hirsutus Alouatta seniculus KR902387.1 Point 6 96.45 507 99 1E-139

The finding that Ps. hirsutus hirsutus and Ps. davisi feed on wild vertebrates may help to improve our understanding of the hematophagous habits of these species as well as our understanding of the Leishmania transmission cycle in this region. The presence of H. sapiens DNA in engorged sand flies combined with the high abundance of these species indicates that humans have been exposed to bites from potential vectors in four of the eight localities in Ji-Paraná. Recently, Pereira Júnior et al. (2019) identified the DNA of humans and anteaters in sand flies from forest and peridomicile areas in Rondônia. Our results corroborate their findings and demonstrate that sand flies use a variety of blood meal sources, including humans, in Ji-Paraná; these findings increase our epidemiological knowledge of CL in the region.

(With 1 figure)

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Received: September 02, 2019; Accepted: November 06, 2019

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