t i l Notes on Acanthaspis quinquespinosa Complex (Hemiptera: Reduviidae: Reduviinae) with Description of a New Species from Pakistan

Acanthaspis Amyot and Serville, 1843, is the second-largest genus within the subfamily Reduviinae. Acanthaspis quinquespinosa (Fabricius, 1781), is a common species in Pakistan. Due to its variable color patterns, its taxonomic distinction from Acanthaspis flavipes Stål, 1855 has been confounded. The genitalia in Reduviidae are commonly used for species identification and in present studies, because of identical genitalia, A. flavipes Stål, 1855 stat. restit. is restored as a junior synonym of A. quinquespinosa . The authors surveyed different locations of Pakistan for members of Reduviinae. In this context, Acanthaspis rafiqi Shah and Cai sp. nov., is described, including descriptions of genitalia and detailed illustrations giving important diagnostic characters. Interestingly, this species has relatively similar body size and color patterns including the structure of male pygophore and parameres, however, differs from A. quinquespinosa in the structure of the phallus.


INTRODUCTION
T he subfamily Reduviinae is polyphyletic (Hwang and Weirauch, 2012) and one of the most heterogeneous within Reduviidae, including more than 1,070 species under the 141 genera (Melo, 2007). The members of this subfamily are cosmopolitan, reaching their greatest diversity in the old and new world tropics (Schuh and Slater, 1995;Weirauch et al., 2014). The genus Acanthaspis was established by Amyot and Serville (1843) for the type species Acanthaspis sexguttata (Fabricius, 1775) and currently it is the second-largest genus in Reduviinae (Hwang and Weirauch, 2012), with 124 species (ITIS, 2020). Ambrose (2006) listed 98 species in 25 genera of Reduviinae in the Indian checklist of assassin bugs, of which 42 belonged to Acanthaspis.
Acanthaspis quinquespinosa (Fabricius, 1781) is a crepuscular, multivoltine assassin bug (Sahayaraj, 1991), and is the common species of this genus in Pakistan. This species can be easily recognized by its black or piceousblack habitus with light to dark yellowish or orangish markings and castaneous legs. However, as the color O n l i n e

F i r s t A r t i c l e
Islamabad, the National Insect Museum (NIM) at National Agricultural Research Center (NARC), Islamabad, Pakistan and Entomological Museum, China Agricultural University (CAU), Beijing. The external morphological structures were examined by using the Nikon SMZ-745 dissecting microscope. The males pygophores i.e. PMNH-59294, PMNH-59307 and, NIMISB-10020 (previously known as A. flavipes), PMNH-59300, NIMISB-10018 and, NIMISB-10023 from A. quinquespinosa, whereas PMNH-59292, and CAUBJ-1005 from Acanthaspis rafiqi Shah and Cai sp. nov. were extracted, soaked in lactic acid for six hours at 28°C, boiled with 20% lactic acid for 20 minutes, rinsed in distilled water and finally, the internal genitalia were extracted from these pygophores under the microscope. The studied genitalia were placed in vials with glycerol and pinned under the corresponding specimens. Photographs were captured by using Canon 7D Mark II digital camera on Olympus BX51 fluorescent and Olympus SZX7 microscopes. The stacking of images was done with Helicon Focus (version 5.3, method C). The measurements were obtained using a calibrated micrometer eyepiece. Morphological terminology mainly follows that of Distant (1904) and Cao et al. (2014).

Distribution
Indomalayan and Afrotropical regions.

Redescription
Coloration: Habitus black to piceous-black with light to dark yellowish or orangish markings; first antennal segment, labium, spot next to ocelli and legs (except coxae and tarsi) castaneous or dark reddish brown (femora medially dark-colored); eyes, antennal segments II-IV, neck, posterior area of prosternum, proacetabula, thoracicsterna and coxae dark to light chocolate-brown; ocelli and tarsi light golden or pale; head, pronotum (excluding area around humeral angles, lateral posterior margins and spines or tubercles at posterior lobe) scutellum and thoracic-pleura black to piceous-black; in male area around the humeral angles, spines or tubercles and lateral impressions of posterior pronotal lobe with a series of light to dark yellowish color (Fig. 5A, C-D), in female, corresponding areas light to dark orangish ( Fig. 6A-C), however, color markings and their patterns can be variable; abdomen with combination of chocolate-brown and dark black to piceous-black; in male connexivum dark black to piceous-black and yellowish (Fig. 8A, B), while, in female piceous-black and dark orangish (Fig. 8C, D); at the base of hemelytra, a small spot, corium with a large light to dark yellowish or sometimes white to creamy spot (Figs. 1A, 2A, 3A, 4A, C) approaching to costal margin; corium slightly dark chocolate-brown while, with membrane light brown; hindwings mostly white with robust brownish veins (Fig. 7B).

F i r s t A r t i c l e
Notes on Acanthaspis quinquespinosa Complex 5 scutellum clothed with reddish-brown or ochraceous, short and long, erect and slightly procumbent stiff setae and long hairs; apically pedicel, flagellomeres and tip of stridulitrum amalgamated with chocolate-brown short stiff setae and long hairs; maxillary plates, thoracic-pleura and ventrally abdomen infested with short adpressed creamywhite or light-brown setae while, most of the dorsum of abdomens glabrous; anterior margin of the collar with stiff erect light-brown setae; antennal sockets posteriorly bearing few stiff bristles; ventrally pro-trochanters and pro-and mesofemora densely furnished with reddishbrown long pilose whereas, pilosity of pro-, meso-and metatibiae short. Structure: Head oblong, slightly longer than first antennal segment; length of anteocular region shorter than postocular region including neck; neck subequal in length with anteocular region whereas, its greatest width individually subequal with basal two labial segments; mandibular plates slightly higher than clypeus (Fig. 5B); maxillary plates feebly rugose; gula smooth shiny; medial lobe of head divided into two plates ( Fig. 5A) separated by interocular sulcus close to mandibular plates; eyes reniform, protruding; ocelli distinctly protuberant ( Fig. 5A-B), slightly apart from each other; neck much distinct; rostrum robust, first visible segment subequal to the second visible segment, medial longitudinal groove very prominent; first antennal segment cylindrical, thirdlongest followed by second.
Male hemelytra of slightly surpassing abdominal tip while, female nearly reaching it, hemelytron at base narrowed, apical margin short, anal margin distinctly long, membrane oval, clavus distinct (Fig. 7A) and outer cell slightly larger than inner cell; anterior margin of hindwing straight, secondary veins (SV) prominently "V-shaped" (Fig. 7B); procoxae longer than meso-and metacoxae; mesofemora less incrassate than pre-femora and more from metafemora; pro-and mesotibiae with fossula spongiosae; metatibiae in male 1.76 times longer than protibiae and 1.58 times than mesotibiae whereas, in female metatibiae 1.73 times than protibiae and 1.57 times from mesotibiae.

Diagnosis
Habitus similar to A. quinquespinosa however, it can be easily differentiated with the following; head distinctly longer than antennal scape; no spines or tubercles at the posterior lobe of pronotum, typically furnished with four spots (Figs. 11, 12, 13C-D); antennae (Fig. 13B), labium (Fig. 13A) and legs (Fig. 15A-D) are light to dark brownish; scent glands in female situated at third tergite while, fourth indistinctly present and fifth reduced (Fig.  16A); dorsal phallothecal sclerite short (Fig. 17A, C) with its apex and basal marginal areas are serrate (Fig. 17A, C), basal, medial, dorsal lobe of the endosoma smaller than A. quinquespinosa.

Description
Coloration: Generally body black to piceous-black and dark chocolate-brown; first antennal and labial segments, a spot next to ocellar area and legs (excluding tibiae and tarsi) dark chocolate-brown; antennal segments II-IV, thoracic-sterna, membrane of hemelytra and ventrally abdomen brown; second and third labial segments light brownish, tarsi ochraceous and tibiae brownish; dorsum of abdomen piceous-black with brown-tinged; head, stridulitrum, pronotum (excluding posterior pronotal lobe posterior area and humeral spines) and thoracic-pleura piceous-black; humeral spines and posteriorly posterior lobe of pronotum furnished with four beige color spots (in male light beige, Fig. 13C while, in female with slightly darker beige, Fig. 13D); basal spot of hemelytra and spot to corium light pale in male whereas, in female it darker O n l i n e

F i r s t A r t i c l e
Notes on Acanthaspis quinquespinosa Complex pale; hindwings white with soil type brownish veins (Fig.  14C), connexiva precisely with combination of pale, piceous-black and dark chocolate-brown. Fig. 11. Acanthaspis rafiqi Shah and Cai, sp. nov., ♂ habitus. Scale bar: 3.00 mm. Vestiture: Male and female both macropterous; body medium-sized, generally elliptic (Figs. 11, 12) and heavily ochraceous pilose; clypeus, area posterior to antennifers, gena, labium, lateral margins of pronotum, basal and lateral margins of coria, scutellum, apical tip of prosternum, connexival margins and ventrally abdomen conflated with short and long, erect and decumbent hairs but, dorsum of abdomen mostly glabrous; antennae with short stiff setae; thoracic-sterna and gula smooth shiny; protrochanters densely clothed with short hairs while, pilosity of pro and mesofemora more than metafemora and pro-, meso-and metatibiae longer pilose.
Structure: Head oblong, oviform and distinctly longer than the first antennal segment whereas, its greatest width individually subequal with basal two labial segments; length of anteocular region smaller than the postocular region including neck (Fig. 13A, B) while, its neck equal or subequal in length with anteocular region; mandibular plates elevated than clypeus; maxillary plates and area of buccula distinctly separated from gula; medial lobe of head divided into two promising sclerotized plates separated by a distinct interocular suture extended to mandibular plates (Fig. 13B), each plate bearing a transverse dark chocolatebrown spot; antennifers tuberculate; eyes reniform, protruding outside; ocelli protuberant, occurring apart from each other (Fig. 13B); rostrum robust, first visible segment subequal to second visible segment, slightly curved, (Fig. 13A) and medially split with a longitudinal groove; first antennal segment cylindrical.
Collar process small but, distinct; male posterior pronotal lobe 1.92 times wider than anterior pronotal lobe while, in case of female it is 1.96 times; anterior lobe of pronotum strongly sculptured and elevated whereas, posterior lobe rugose and slightly declivous anteriorly; humeral angles spinously produced (Fig. 13C, D); posterior pronotal lobe furnished with four spots, two to each close to humeral spines and two posteriorly adjacent to humeral spots, each of posterior spot medially bearing a very indistinct small tubercle (sometimes might be reduced) or much acute indistinct spines; scutellum "Y-shaped", its spine inclined to the abdomen; proepisternum and mesopleura smooth while, proepimeron and metapleura feebly rugose; proacetabula sulcated while, meso-and metaacetabula mostly smooth; stridulitrum "V-shaped" with 1.5 mm long, deep prosternal groove (Fig. 14B), its apical tip almost extending beyond the procoxal cavities.
Hemelytra of male and female not surpassing abdominal tip; outer cell of hemelytron rectangular, inner cell ovate, basal junction bearing a small indistinct spot, apically corium with a large round spot approaching anterior margin; clavus distinct (Fig. 14A); hindwing anterior margin straight (Fig. 14C), humus short but distinct, secondary veins distinctly separated; pro-coxae about two times as long as wide; meso-femora less incrassate than profemora and more from metafemora (Fig. 15); pro-and mesotibiae with spongy furrows (  Female abdomen oblong, ovoid (Fig. 16A, B), second tergite subdivided into three subparts, connexivum conspicuously exposed at repose; ventrally central ridge in male abdomen prominent than female; in female, scent glands situated at third tergite while, fourth indistinct, seventh tergite concave, eighth narrowed and convex, ninth and tenth triangular and distinctly fused (Fig. 16C, D), first valvifer plates triangular with anterior margin convex and posterior margin straight, first valvula indistinct than first valvifer and styloid long pointed.

Etymology
This new species is named in honor of Muhammad Rafiq, former head of Entomology Section and Principal Scientific Officer, Central Cotton Research Institute, Multan, Punjab, Pakistan for his long distinguished and superior service to the institution.

Biology
The holotype was collected at night from the boulderclay and stones (Fig. 18A, B), which was nearby covered with dense natural vegetation and trees. The paratype female was collected close to the male holotype, while, one paratype male was collected from a dead tree trunk at early night time.
Lateral and basal spines of pronotal regions of type A. flavipes (Fig. 4C, D), A. quinquespinosa and A. flavipes illustrated by Chandra et al. (2012) including the image of A. quinquespinosa (UCR-ENT-00014950, India, 1949) uploaded on http://research.amnh.org/pbi/ heteropteraspeciespage/speciesdescriptionall.php as well as specimens collected by us (Figs. 5A, C, D, 6A,C) had a series of light to dark yellowish or dark orangish color patches. Whereas, syntype A. quinquespinosa (Fig. 4A, B) and the similar species illustrated by Cao et al. (2014) and Chandra et al. (2014) had a spot around the lateral and basal spines. Almost similar findings are mentioned by Cao et al. (2014) regarding the coloration, who recorded six kinds of variations on posterior lobe of pronotum in Chinese individuals of Acanthaspis cincticrus Stål (1859), and those of Ambrose and Livingstone (1987), and Sahayaraj (2007), who observed color polymorphism and different ecotypes of the genus.
In accordance with images of types species and key provided by Distant (1904), the specimens of A. quinquespinosa complex were grouped based on spines (Fig. 5A, C), tubercles (distinct/indistinct, Figs. 5D, 6A, B) and tubercles surmounted with very short tips (Fig.  6C). A pair of spines at pronotal lobe in Acanthaspis quinquespinosa were mentioned by Distant (1904), Afzal (2005), Khot and Hegde (2010), Cao et al. (2014) and, Mukherjee (2015), whereas, A. flavipes was mentioned with discal tubercles (Distant, 1904;Afzal, 2005). Moreover, distinct tubercles were observed in specimens of A. flavipes (collected by M.S.K., a data label preserved at the entomological museum, Center for Agriculture and Biosciences International (CABI), Rawalpindi, Pakistan. The specimens having distinct/indistinct tubercles or tubercles surmounted with short tips had genitalia identical to A. quinquespinosa. The dissected genitalia of A. quinquespinosa and A. flavipes completely match with those illustrated by Cao et al. (2014). Therefore, we strongly agree with Maldonado Capriles (1990), Ambrose (2006) and Afzal (2005), who followed Stål (1874), about synonymy of A. flavipes with A. quinquespinosa. We could not find any other differences in morphological characters and thus, A. flavipes Stål (1855) stat. restit. is restored as a junior synonym of A. quinquespinosa.
Moreover, Acanthaspis rafiqi Shah and Cai sp. nov., the pygophore and parameres are the same in structure with A. quinquespinosa, but differ in the structure of the phallus. The pedicel of A. quinquespinosa is split and curved (Fig. 10B, C), while in A. rafiqi, it is fused, slightly curved and thickened (Fig. 17B). The dorsal phallothecal sclerite of A. quinquespinosa is long (Fig. 10A, D), medial apical process apically concave and fused with strut; posterior to the dorsal phallothecal sclerite, a club-shaped basal, medial dorsal lobe of the endosoma (Fig. 10A, D). In the case of A. rafiqi, the dorsal phallothecal sclerite is short (Fig. 17A, C), its apex and marginal areas over the strut are serrate (Fig. 17A, C) and sclerotized. The basal, medial, dorsal lobe of the endosoma is 5.45 times smaller than that of A. quinquespinosa. Fig. 19. Distribution of Acanthaspis quinquespinosa (Fabricius, 1781) and A. rafiqi Shah and Cai, sp. nov., in various localities of Pakistan.

ACKNOWLEDGMENTS
We are grateful to Dr. Khalid Abdullah (Cotton O n l i n e