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

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)

References

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Publication Dates

• Publication in this collection
  17 Feb 2020
• Date of issue
  Jan-Feb 2021

History

• Received
  02 Sept 2019
• Accepted
  06 Nov 2019