Research Article |
Corresponding author: João Aristeu da Rosa ( rosaja@fcfar.unesp.br ) Academic editor: Guanyang Zhang
© 2017 João Aristeu da Rosa, Hernany Henrique Garcia Justino, Juliana Damieli Nascimento, Vagner José Mendonça, Claudia Solano Rocha, Danila Blanco de Carvalho, Rossana Falcone, Maria Tercília Vilela de Azeredo-Oliveira, Kaio Cesar Chaboli Alevi, Jader de Oliveira.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Rosa JA, Justino HHG, Nascimento JD, Mendonça VJ, Rocha CS, Carvalho DB, Falcone R, Azeredo-Oliveira MTV, Alevi KCC, Oliveira J (2017) A new species of Rhodnius from Brazil (Hemiptera, Reduviidae, Triatominae). ZooKeys 675: 1-25. https://doi.org/10.3897/zookeys.675.12024
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A colony was formed from eggs of a Rhodnius sp. female collected in Taquarussu, Mato Grosso do Sul, Brazil, and its specimens were used to describe R. taquarussuensis sp. n. This species is similar to R. neglectus, but distinct characters were observed on the head, thorax, abdomen, female external genitalia and male genitalia. Chromosomal differences between the two species were also established.
Brazil, cytotaxonomy, new species, Rhodnius , taxonomy, Triatominae
In the subfamily Triatominae, the genera Panstrongylus (15 species), Triatoma (74 species) and Rhodnius (20 species) are of particular epidemiological importance, although the other 15 genera (containing 43 species) can also transmit Trypanosoma cruzi, which is the etiological agent of Chagas disease (
The first two species identified as belonging to the genus Rhodnius were described by
Most Rhodnius species live in palm trees, and several cases of transmission of Chagas disease have been associated with the consumption of açaí containing feces of triatomines infected by T. cruzi (Ferreira, Branquinho & Leite, 2014;
Based on morphological, morphometric and cytogenetic characters, this paper describes R. taquarussuensis sp. n., which is similar to R. neglectus. The first collected specimen of R. taquarussuensis was a female that invaded a domicile and laid eight eggs. The colony formed from those eggs resulted in the specimens used in this description.
On 10 November 2010 a female of Rhodnius sp. invaded a rural dwelling (22°29'07.7"S; 53°21'08.9'W) in the city of Taquarussu, Mato Grosso do Sul, Brazil, and was captured (Fig.
A colony was formed from the eight eggs laid by the R. taquarussuensis sp. n. female and identified as Araraquara Triatominae Colony (CTA) 277. The specimens of that colony were used to describe R. taquarussuensis sp. n.
The morphological study by OM and scanning electron microscopy (SEM) consisted of the observation of the head, thorax and abdomen of 30 adult females and 30 adult males, as well as 40 eggs of R. taquarussuensis sp. n. and the same number of specimens of R. neglectus, according to
Female external genitalia were observed from the dorsal, posterior, and ventral sides (Fig.
The Leica MZ APO stereoscope from the Faculty of Pharmaceutical Sciences, UNESP, Araraquara, and the scanning electron microscope Topcon SM-300 located in the Department of Physical Chemistry at the Chemistry Institute, UNESP, Araraquara, were used for observation and capture of images.
In the morphometric study by OM, 15 egg shells, 15 females and 15 males from the colony were measured, the same being done for R. neglectus CTA 229 (Table
Mean of measurement (mm) of 15 females and 15 males of R. taquarussuensis sp. n. and R. neglectus.
Female | Male | |||
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R. taquarussuensis | R. neglectus | R. taquarussuensis | R. neglectus | |
TL | 17,25 | 17,25 | 15,24 | 15,96 |
MLA | 9,86 | 10,03 | 8,41 | 9,05 |
MLT | 5,00 | 4,11 | 4,54 | 3,79 |
R1 | 0,75 | 0,92 | 0,71 | 0,92 |
R2 | 3,38 | 3,58 | 3,16 | 3,59 |
R3 | 0,85 | 0,95 | 0,77 | 0,96 |
HL | 4.44 | 5,81 | 4.20 | 5,24 |
EO | 1.53 | 1,98 | 1.42 | 1,80 |
IE | 0.61 | 0,77 | 0.54 | 0,66 |
PO | 1.00 | 3,67 | 1.04 | 3,37 |
AO | 2.62 | 0,90 | 2.44 | 0,91 |
AT | 1.93 | 2,30 | 1.82 | 2,30 |
SC | 1,96 | 2,03 | 1,69 | 1,86 |
A1 | 0,45 | 0,55 | 0,29 | 0,59 |
A2 | 3,56 | 4,13 | 2,24 | 4,35 |
A3 | 2,03 | 2,43 | 1,28 | 2,50 |
A4 | 1,36 | 1,82 | 0,87 | 1,92 |
Eggs | R. taquarussuensis | R. neglectus | ||
TE | 1,72 | 1,62 | ||
OO | 0,49 | 0,52 |
The observations and measurements were carried out on a Leica MZ APO stereoscope and the Motic Images Advanced System version 3.2.
In this study ten male specimens of R. taquarussuensis sp. n. were used for C and CMA3/DAPI–banding analyses and ten male specimens of R. neglectus were used for CMA3/DAPI–banding analyses. After being lacerated and placed on the slide, the seminiferous tubules underwent cytogenetic procedures following the C-banding (
BRAZIL: Mato Grosso do Sul: Taquarussu; Residence, 22°29'07.7"S; 53°21'08.9'W, 10 November 2010 H. E. G. Justino. UNESP (♀).
BRAZIL: Colony formed from eggs obtained from the holotype: Araraquara: Triatominae Insectarium of the Faculty of Pharmaceutical Sciences, Araraquara, January 3, 2017, J. A. da Rosa, UNESP (25 ♂ 25 ♀).
CTIOC - Collection of Triatomines of the Oswaldo Cruz Institute, Rio de Janeiro - Brazil (2 ♂ 2 ♀). Entomological Reference Collection of the Faculty of Public Health - USP, São Paulo -Brazil (1 ♂ 1 ♀). Collection of the Institute of Entomology of the Metropolitan University of Education Sciences (IEUMCE), Santiago - Chile (2 ♂ 2 ♀).
The name Rhodnius taquarussuensis sp. n. was chosen because this species was found in the city of Taquarussu, Mato Grosso do Sul, Brazil.
Rhodnius taquarussuensis sp. n. is close to R. neglectus, their differences being the color and a variety of morphological, morphometric and cytogenetic characters (Tables
Main distinguishing characters between R. taquarussuensis sp. n. and R. neglectus.
Distinguishing characters | Species | ||
---|---|---|---|
R. taquarussuensis | R. neglectus | ||
Overall color | Brown | Dark brown | |
Genae | Lengthier longer | Longer | |
Vertex | Quite visible | Not visible | |
Ventral triangular furrow | Filamentous way | Rounded way | |
Scutellum | Covers the final portion of the urotergite I process | The apex of the process of the urotergite I is perfectly visible | |
Stridulatory sulcus | Straight | Waisted | |
Mesothorax | Half-moon shaped and regular | Pronounced and slightly irregular | |
Female external genitalia | Dorsal side | 10th segment presents a concavity in the middle portion | 10th segment is straight |
Posterior side | The limits of the 9th segment with gonocoxite VIII are curve | The limits of the 9th segment with gonocoxite VIII are straight | |
Ventral side | There is a concavity in the external limit with the 10th segment | There is a straight line in the external limit with the 10th segment | |
Male genitalia | Phallothecal sclerite | Trapezoidal shape | Rounded shape |
Tip of parameres | Thinner | Thin | |
Heterochromatin in the autosomes | Present | Absent | |
CMA+ in autosomes | Present | Absent |
On the head, differences were noticed on the vertex, genae, antennae and triangular furrow of the first segment of the rostrum. The vertex of the head of R. taquarussuensis sp. n. is quite visible, whereas on R. neglectus it is not (Fig.
Among the 19 characters measured, 12 showed significant differences between R. taquarussuensis sp. n. and R. neglectus in both sexes and also the eggs of both species. Two characters showed differences only between males, and five characters did not show significant differences (Tables
A total of 15 adult females and 15 adult males of R. taquarussuensis sp. n. and R. neglectus were measured, as well as 30 eggs shells of both species. Such measurements are detailed in Table
The head of R. taquarussuensis sp. n. has a prominent brown vertex contrasting with the black sides. The clypeus is well defined. The genae are large, visible and dark brown, moving towards the anteclypeus (Figs
The first segment of the antennae is black with mixes of brown. The articulation between the first and second segment of the antennae is brown. Roughly all the 10th part of the beginning of the second antennal segment is brown. The second segment is mostly black. In the articulation between the second and third antennal segment there is a black ring followed by a brown one. The beginning of the third segment (around 1/3) is black and the remaining portions (2/3) are brown. The articulation between the third and fourth antennal segment is brown. The beginning of the fourth segment is black and the remaining portions are brown with mixes of black (Fig.
The eyes are black and the ocelli are brown. The neck has a brown central dorsal strip flanked by two (1+1) black, narrower strips. The ventral portion of the neck between the ocelli is dark brown (Fig.
The pronotum of the thorax of R. taquarussuensis sp. n. has a trapezoidal shape and is limited by a brown carina. In the antero posterior direction the pronotum has other two brown carina in the middle portion and six black strips. The three carina and the three brown strips are interspersed with the six black strips, which are larger. The collar (first portion of the pronotum) in the central part is brown and is followed by two (1+1) black glabrous areas and the two (1+1) antero lateral angles. The anterior portion of the pronotum consists of three anterior lobes which are clearly distinct from the posterior portion (hindlobe). Those three anterior lobes are limited by the carina and on each of them there are two black glabrous areas with a lengthy and irregular outline (Fig.
The cuticle involving the veins of the hemelytron is light brown. The corium between the veins of the coriaceous region is dark brown, whereas that of the membrane is brown (Fig.
The prosternum contains the stridulatory sulcus, which moves along that segment in an antero-posterior direction, having a brown color in the background and black on the sides. Two elongated tubercles limit the anterior half of the stridulatory sulcus. In the superior portion and in diagonal direction from the tubercles there are two black glabrous areas surrounded by a set of brown sensilla (Fig.
The mesosternum is limited anteriorly by the prosternum and posteriorly by the metasternum, both limits being brown. The central line dividing two dark brown elevations is also brown. Those two elevations are limited by two (1+ 1) black side glabrous areas diagonally placed. The central region of the posterior limit of the mesosternum has a half-moon shape. The metasternum is brown and resembles an isosceles triangle. Its anterior portion, i.e., its limit with the mesosternum, corresponds to the vertex of the triangle and is narrow, whereas its posterior portion, i.e., its limit with the first abdominal segment, corresponds to the basis of the triangle (Fig.
The three pairs of coxae are brown, except for the black glabrous areas. The trochanters of the anterior pair of legs are brown, but mixed with black glabrous areas. The middle and posterior pairs of trochanters are brown and have no glabrous areas. The three pairs of femora are black and the same color prevails in the three pairs of tibiae, except in the articulations with the femur and the spongy fossula, which are brown. The spongy fossulae are located in the first and second pairs of legs in the final portion of the tibia, alongside the articulations with the tarsi (Fig.
The abdomen of R. taquarussuensis sp. n. presents a brown color in the longitudinal central portion. On the sides of each segment there are (3+3) black glabrous areas, which are mixed with brown and black areas. The connexivum of the dorsal portion lies between the second and seventh segment. For each of those segments the anterior half is black and the posterior one is brown. The dorsal connexivum, also lying between the second and seventh segment, has a black color in 2/3 of the anterior portion, but that black color ends in an irregular way over the remaining 1/3, which is brown. Therefore, the black portion of the connexivum presents two edges moving towards the brown portion: one in the internal limit of the connexivum and the other in the middle portion. However, the connexivum of the second dorsal segment is black in the anterior half and brown in the posterior one, the limit between the portions having a diagonal shape. The seventh segment, on the other hand, is practically all black, except for a small brown strip located in the external posterior half. Type 1 sensilla, which prevail on the head, thorax and abdomen, have a brown color (Fig.
Male genitalia have the typical aspect of the genus Rhodnius. The median process of the pygophore (PrP) is short and triangular, but the base is broad and the sides are elongated with a thin edge. Parameres are hairy with a thin edge. From ventral view, the phallosome (Ph) has a broad plate whose superior region has a trapezoidal shape and occupies the middle region of the aedeagus. The support of the phallosome plate (PrPh) is broad. Conjunctival process I (PrcjI) is present and II (PrcjII) is absent. Endosomal process (En) is well-developed when seen from dorsal and ventral view (Figs
Phallus of R. taquarussuensis sp. n. A dorsal view C ventral view, R. neglectus B dorsal view D ventral view. Cj: conjunctive, En: endosome, EPlb: median extencion of basal plate, P: phallus, Plb: basal plate, PrG: gonopore process, PrPh: phallossoma process, Ph: phallosoma, PrCj: conjunctive process, ll: line limit.
The dorsal side of the female external genitalia presents a concavity in the middle portion of the 10th segment. Seen from posterior view, the limits (1+1) of the 9th segment with gonocoxite VIII are curve, whereas the superior line limiting the 10thand 9thsegments is straight. In the central portion of the 10th segment of the ventral side of the female external genitalia there is another concavity that can be noticed from dorsal view. The external limits (1+1) of the 9th segment of the female external genitalia are curve when seen from ventral view (Fig.
Egg shells of R. taquarussuensis sp. n. have a length of 1.72 mm and an opercular opening of 0.49 mm. They present lateral flattening, collar and exochorion cells, most with pentagonal or hexagonal shape (Fig.
Finally, although R. taquarussuensis sp. n. showed the same number of chromosomes as R. neglectus and all the tribe Rhodniini, i.e., 2n = 22 (Figure
A Right wing of R. taquarussuensis sp. n. with the seven landmarks used in morphometric analysis. Following Bookstein (1990), all points correspond to type I landmarks (venation intersections) B Factorial maps in the plane of the two discriminant factors of wing shape variation (canonical variables 1 and 2, or CV1 and CV2) presenting the distribution of specimens of R. taquarussuensis sp. n. (Rta, black cicle) and R. neglectus (Rne, silver cicle).
Constitutive heterochromatin pattern in R. taquarussuensis. A Initial prophases with a chromocenter heterochromatic consisting of sex chromosomes (arrow) and several heterochromatic blocks dispersed in the nucleus B Metaphase I with heterochromatic blocks in both extremities of practically all the autosomes and in the Y sex chromosome. X: X sex chromosome, Y: Y sex chromosome. Bar: 10 μm.
Composition of the pairs of bases of DNA rich in AT and CG in R. neglectus (A, B) and R. taquarussuensis (C, D). A X sex chromosome rich in CG B Y sex chromosome rich in ATC X sex chromosome and various blocks dispersed in the prophase nucleus (arrows) rich in CG D Y sex chromosome rich in AT. X: X sex chromosome, Y: Y sex chromosome. Bar: 10μm.
The subfamily Triatominae include 18 genera comprising 152 species, 20 of which belong to the genus Rhodnius (
In addition to the macroscopic characters, R. taquarussuensis sp. n. and R. neglectus were considered “close to” because the OM study indicated similar characters between them, including: placement of black and brown spots on the dorsal and ventral connexivum, length of the four segments of the antenna, pronotum, antero lateral angles, urotergite I process, geometric morphometry of the hind wings, median process of the pygophore and morphological characters of the eggs.
Rhodnius taquarussuensis sp. n. was considered distinct from R. neglectus on account of the observation of color, eleven morphological characters, twelve morphometric characters and cytogenetic features (Tables
Regarding the color, the distinction between R. taquarussuensis sp. n. and R. neglectus was based on the general aspect, segments of the antenna, hind wings and stridulatory sulcus. The general color of R. taquarussuensis sp. n. is brown, whereas R. neglectus is dark brown, almost black, or “brown dark”, as referred to by
Out of the eleven morphological characters that distinguish R. taquarussuensis sp. n. from R. neglectus, three are located on the head: dorsal vertex, genae and triangular furrow of the first segment of the proboscis. The difference related to the vertex was one of the characters used by
In what refers to the thorax, differences on the scutellum, protothorax, mesothorax, and metathorax were noticed. The scutellum of R. taquarussuensis sp. n. and R. neglectus differs in the shape of the apex and also the position on which that apex reaches urotergite I, since there is no significant difference in terms of length (Table
The differentiation of seven genera of triatomines based on the shape of the prosternal stridulatory sulcus was carried out by
The eggs of R. taquarussuensis sp. n. and R. neglectus showed differences in the measurement of their length and opercular opening, the same as R. montenegrensis and R. marabaensis on the occasion of their description. As for the morphological characters, no differences were recorded. On the other hand, it should be noted that morphological differences were found by
According to Justi and Galvão (2016) the group R. prolixus comprise the following species: R. batretti, R. dalessandroi, R. domesticus, R. marabaensis, R. milesi, R. montenegrensis, R. nasutus, R. neglectus, R. neivai, R. prolixus and R. robustus. Since R. taquarussuensis sp. n. is close to R. neglectus we suggest the inclusion of R. taquarussuensis sp. n. in the R. prolixus group and we present the main differences between the twelve species (Table
Distinguishing characters among twelve species of the group Rhodnius prolixus.
Species | Distinctive characters | References |
---|---|---|
R. barretti | The third antennal segment appears to be relatively shorter. The scutellar process is narrowly pointed. |
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R. dalessandroi | Antenniferous tubercle slightly pilose and with triangular glabrous depression in the upper region. Semicircular spot on the posterior end of the neck. | Carcavallo and Barreto 1976 |
R. domesticus | Head comparatively long, distinctly longer that pronotum. Process of pygophore rectangular. |
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R. marabaensis | The second antennal segment is 10.3 times larger than the first. The scutellum is larger and includes two prominent internal lateral carinea. |
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R. milesi | The male genitalia presents a second process of the phallosoma. Divergent antennal tubercle with an apical denticle. | Valente et al. 2001 |
R. montenegrensis | Anterior wings with well-demarcated veins, notable the Sc by a yellow tonality. Abdomen presents yellow spots interposed with dark ones over the ventral abdomen lengthwise. |
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R. nasutus | Overall color light reddish brown, trochantera not contrasting conspicuously with femora. Median process of pygophore wide at base. |
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R. neglectus | Overall color dark brown, trochantera very light colored. Median process of pygophore narrow at base. |
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R. neivai | Pronotum entirely dark brown or black, including the carine. Connexivum blackish, with very small reddish spots. |
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R. prolixus | Anteocular region slightly over three times as long as postocular. Distance between eyes dorsally larger than width of eyes in dorsal view. |
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R. robustus | Anteocular region about four times as long as postocular. Specimens distance between eyes dorsally smaller than, or equal to, width of eye in dorsal view. |
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R. taquarussuensis sp. n. | Head with a prominent brown vertex contrasting with the black sides. The phallosome (Ph) has a broad plate whose superior region has a trapezoidal shape and occupies the middle region of the aedeagus. | This work |
Cytogenetic analyses of R. taquarussuensis sp. n. made it possible to describe the karyotype (2n = 22) and observe the constitutive heterochromatin pattern in the chromosomes (extremities of most autosomes), which are rich in CG. All the species in the tribe Rhodniini have 22 chromosomes (
Although the evolutionary process in triatomine is disruptive (
Conceived the study: JAR, HHGJ, JO and KCCA.Colected the bugs:HHGJ.
Prepared samples: JAR, JDN, JO, DBC and JDN. Analysed data: JAR, JO,VJM, RF,CSR and MTAO. Interpreted data: JAR, JO, KCCA, DBC. Wrote the manuscript: JAR, JO and KCCA.
All authors read and approved the final version of the manuscript.
Marcelo Ornaghi Orlandi, Mario Cilense e Sebastião Dametto from the Institute of Chemistry at São Paulo State University (UNESP) by the support and help regarding the use of the scanning electron microscope. Heloisa Pinotti, Eder dos Santos Souza, Lucas Abrantes da Silva, Tiago Belintani, Fábio Regis Garcia postgraduates of the Bioscience and Biotechnology Applied to Pharmacy program (FCF / UNESP) by the constant help in the maintenance of triatomine colonies. Nelson Papavero from the Zoological Museum of USP and Hélcio Reinaldo Gil-Santana of Diptera Laboratory, Oswaldo Cruz Institute for suggesting the scientific name of the new species, to Dennys Ortiz postgraduate of Animal Biology (UNICAMP) for the support with maps, to Davi Antonio da Rosa for the photos. The authors are also very grateful to Dr. Guanyang Zhang (Editor of Zookeys), Dr. Daniel A. Frias Lasserre (Laboratory of Entomology, Metropolitan University of Education Sciences, Santiago, Chile), Jean-Michel Bérenger (Laboratory Diagnostic Insect, Bouc-Bel-Air, France) and an anonymous reviewer for their valuable comments and suggestions. Financial Support: Scientific Support and Development Program of School of Pharmaceutical Sciences (UNESP);São Paulo Research Foundation (FAPESP) grant process number 2009/52236-2 and 2013/19764-0; Brazilian Federal Agency for the Support and Evaluation of Graduate Education (CAPES), grant process number 23038-005285/2011-2012; Brazilian National Council for Scientific Technological Development (CNPq process number 142284/2015-7).