Turkish Journal of Zoology Turk J Zool (2015) 39: 1034-1040 http://journals.tubitak.gov.tr/zoology/ © TÜBİTAK Research Article doi:10.3906/zoo-1401-17

Taxonomic study of subfamily Scymninae (Coleoptera: Coccinellidae) with one new record from Gilgit-Baltistan, Pakistan

1, 2 1 1 Muhammad ASHFAQUE *, Farman ULLAH , Muhammad Ather RAFI , Falak NAZ 1 Department of Plant and Environmental Protection, National Agricultural Research Center, Islamabad, Pakistan 2 Department of Plant Protection, University of Agriculture, Peshawar, Pakistan

Received: 10.01.2014 Accepted/Published Online: 07.07.2015 Printed: 30.11.2015

Abstract: The fauna of the subfamily Scymninae (Coccinellidae: Coleoptera) of Gilgit-Baltistan was explored and studied. This study confirmed the occurrence of 4 species in 2 genera of Scymninae. These include 2 species in genus Scymnus (S. posticalis Sicard, 1913 and S. nubilus Mulstant, 1850), and 2 species in genus Stethorus (Stethorus gilvifrons (Mulstant, 1850) and S. pauperculus (Weise, 1895)). Among these 4 species, S. posticalis Sicard is reported here as a new record from the study area. A description of each species is given, along with color plates of adult specimens and genitalia. Remarks pertaining to taxonomic discussion, history, taxon status, hosts, habitat, identification key, and distribution of each species in the study area are also given.

Key words: Coccinellidae, Scymninae, fauna, Scymnus, Stethorus, taxonomy, distribution, Gilgit-Baltistan

1. Introduction species from Pakistan and current knowledge on the Coccinellid beetles (Coccinellidae: Coleoptera), commonly predatory coccinellid beetles of Gilgit-Baltistan are mainly known as ladybirds, lady beetles, or ladybugs, are among based on the work of Rafi et al. (2005). In this compilation, the best-known beneficial predatory insects. Many works only 9 species of predatory coccinellids belonging to 3 recognize 6 subfamilies within this family, including subfamilies were listed from this area. Chilocorinae, Coccinellinae, Coccidulinae, Scymninae, Some members of the subfamily Scymninae are Sticholotidinae, and Epilachininae. Of these, only one predacious on tetranychid mites and are known as subfamily (Epilachininae) is phytophagous; the rest are acriphages, while others feed on scale insects and are predacious in nature. Vandenberg (2000) provided a list known as coccidophages. The subfamily is distributed of 6000 species in 370 genera of Coccinellidae worldwide, worldwide. Fürsch (1996) reported 53 genera and 25 while Ślipiński (2007) reported 6000 species in 490 genera. subgenera in 10 tribes, while Kuznetsov (1997) reported They are of economic importance due to their predatory 72 genera in 9 tribes. activity against soft-bodied insects like aphids, jassids, Irshad (2001) reported 71 species of coccinellids from psyllids, whiteflies, scale insects, and mealy bugs. They also Pakistan, but Irshad (2003) reduced this list to 61 species prey on small larvae, insect eggs, and phytophagous mites, by removing the doubtful species. Similarly, Rafi et al. which are injurious to agricultural and forest plantations. (2005) listed 75 species of predatory ladybird beetles from Predation by adults and larvae of the coccinellid beetles Pakistan. All of these compilations are based on external can have a major impact on populations of immature morphology of the adult specimens, which might result in stages of these insects (Rafi et al., 2005). identification mistakes at the species level. Although subfamilies of the Coccinellidae are more or Gilgit-Baltistan lies between 72°N and 75°N and less worldwide in distribution, many species within these between 35°E and 37°E; biogeographically, the area is in subfamilies are restricted to particular biogeographical the Palearctic region and belongs to the dry mountain regions. The Indo-Pakistan subcontinent is known to have subecological zone of Pakistan. The area is highly diverse over 300 species of Coccinellidae (Rahatullah et al., 2011). and climatic conditions vary from extreme cold to Initial documentation of predatory coccinellids of extreme hot spots, with diverse flora and fauna. The area Pakistan was done by Irshad (2001, 2003) and Irshad has many apple and grape plantations, which have mite and Khan (2005). Details regarding predatory coccinellid and scale insect infestations. As already stated, Scymnus * Correspondence: [email protected] 1034 ASHFAQUE et al. / Turk J Zool coccinellids are predators of these pests; therefore, there Male genitalia were mounted onto a plastic strip adhesive is a need to explore the Scymnus coccinellids of Gilgit- with hydrosoluble glue, and the female genitalia onto a Baltistan. Moreover, proper taxonomic studies are strip of transparent plastic with Euparal. necessary for developing biological control-based insect 2.4. Description pest management strategies (Irshad and Haq, 2010). To minimize redundancy, only characteristics peculiar to a given lower-ranking taxon are presented in its description. 2. Materials and methods The species were identified and described following the 2.1. The study area – Gilgit-Baltistan literature of Kapur (1958), Canepari and Milanese (1997), Gilgit-Baltistan (formerly the Northern Areas of Pakistan) Katakura et al. (2001), Inayatullah et al. (2005), and Rafi is spread over an area of 72,496 km2 and is administratively et al. (2005). divided into 7 districts: Astore, Diamer, Gilgit, Ghanche, Ghizer, Hunza-Nager, and Skardu. The area lies between 3. Results 72°N and 75°N and between 34.5°E and 37°E. About 44% The results of the present study revealed that there are 4 of the area is covered by mountains (rocks, wetlands, etc.), species in 2 genera of Scymninae (Coccinellidae) in Gilgit- 50% is alpine pasture, 4% forests, and 1% cultivable waste, Baltistan, Pakistan. Distribution of Scymninae species in while only 1% is under cultivation. Three high mountain the study area is shown in Figure 1. ranges, namely the Himalayas, Karakorum, and Hindu 3.1. Subfamily: Scymninae Mulsant, 1846 Kush, are found in the area. The area is home to 5 of the Synonyms: Scymninae Mulsant, 1846; Scymniens Mulsant, world’s highest peaks of above 8000 m and about 50 peaks 1846; Scymninae Della Beffa, 1912. with elevations of 7000 m. Along with other world-famous 3.2. Diagnosis peaks, Nanga Parbat (known as Killer Mountain) and K2 Body small to medium in size. Eyes and dorsum of (Mount Godwin-Austen) are located in the area. This area body with dense pubescence. Antennae short, 9- to is surrounded by China, India, and Afghanistan, and it is 11-segmented, with distinct club. Terminal segment of separated from Tajikistan by the narrow Wakhan strip. maxillary palpus with truncate apex, cylindrical. Mentum 2.2. Collection and submentum relatively widely articulated. Mandibles For collection of the coccinellid species, surveys were with 1 usually bifid tooth at apex. Pronotum anteriorly conducted in all 7 districts of Gilgit-Baltistan from 2006 to narrowed. Tarsi cryptotetramerous. 2011, mostly in the months of May through September, as 3.3. Key to the genera of subfamily Scymninae of Gilgit- the majority of the areas of Gilgit-Baltistan are accessible Baltistan only during these months due to harsh weather conditions. 1. Pronotum broadly rounded at middle of anterior Each locality was visited fortnightly or monthly depending margin. Last segment of maxillary palpi narrowed ...... upon the accessibility of the area. Standard collection ...... Stethorus procedures were applied. About 120 ladybird beetle 1. Pronotum longitudinally truncate at middle specimens were collected. Field data of each specimen was of anterior margin. Last segment of maxillary palpi recorded. In addition to the fresh collections, the housed cylindrical ...... Scymnus collection of the National Insect Museum (NARC), 3.4. Genus: Scymnus Kugelann, 1794 Islamabad, was also examined. Type species: Scymnus nigrinus Kugelann, 1794, by 2.3. Genitalia extraction and examination subsequent designation of Westwood (1838–1840). For genitalia extraction, the method of Majerus and Synonyms: Scymnus Kugelann, 1794; –Mulsant, 1846; – Kearns (1989) was followed with some modifications. The Crotch, 1874; –Korschefsky, 1931. specimens were softened by putting them into hot water 3.4.1. Scymnus nubilus Mulsant, 1850 for 1 to 2 min. With the help of two entomological needles, Synonyms: Scymnus nubilus Mulsant, 1850; –Bielawski, the abdomen was detached from the body and boiled in 1972; –Booth and Pope, 1989. 10% potassium hydroxide solution for 30 min to dissolve Scymnus (Scymnus) nubilus Korschefsky, 1931. excess tissue and partially clear opaque structures. The Scymnus curtisii Mulsant, 1850 (synonymized by abdomen was cleaned in hot water and transferred to pure Booth and Pope, 1989); Scymnus suturalis Motschulsky, glacial acetic acid for 5 min. To dehydrate the abdomen, 1858; –Crotch, 1874; –Korschefsky, 1931; –Iablokoff- it was first immersed in 80% ethanol and then in absolute Khnzorian, 1972. alcohol for 5 min. Finally, the abdomen was moved to clove Scymnus stabilis Motschulsky, 1866; –Crotch, 1874; – oil. The processed abdomen was put on a cavity slide with Weise, 1900; –Korschefsky, 1931. a drop of glycerin, and the genitalia were extracted with Scymnus lateralis Sicard, 1913; –Korschefsky, 1931; – the help of two entomological needles under a stereoscope. Pang and Gordon, 1984.

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Gulmit

Jaglot

Scymnus nubilus Scymnus posticalis Stethorus gilvifrons Stethorus pauperculus

Figure 1. Distribution of Scymninae species in Gilgit-Baltistan.

Material examined: 3 ♂ 5 ♀ Pak. GB, Gilgit, Jaglot, 22.VI.2006 (Ather) (NIM); 1 ♂ 5 ♀ Pak. GB, Gilgit, Jaglot, 06.VI.2007 (Amjad) (NIM). Diagnosis: Body small, elongate, moderately convex as shown in Figure 2. Head light brown to dark brown. Head deeply inserted, visible from above with eyes large and brown. Pronotum dark brown, darker in middle with lighter reddish brown posterior and anterior margins. Elytra ground color yellowish to reddish brown, punctate with a dark brown to black sutural stripe from basal margin and gradually narrowed toward apex, lateral margins narrowly dark brown to black at middle. Antennae small, segmented. Basal segment is thick, broader than and about twice as long as the second segment. Second segment is as long as and twice as broad as the third segment. Body dark brown from below, densely covered with fine, short, transparent, yellow hairs. Genitalia: Phalobase: Trab long, thick, curved at middle. Basal piece short and rectangular. Parameres thick, slightly longer than median lobe, slightly compressed at base, tips rounded with long sparse hairs. Median lobe broad for most of its length except apex, which is pointed and slightly curved as shown in Figure 3. Figure 2. Adult of Scymnus nubilus.

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Figure 3. Male genitalia of Scymnus nubilus.

Remarks: This species is very similar to S. (Neopullus) Figure 4. Adult of Scymnus posticalis. hoffmanni in general appearance and coloration, and can be differentiated by the male genitalia. Booth and Pope (1989) synonymized S. curtisii Mulsant and S. lateralis Sicard, and gave the valid name of Scymnus nubilus Genitalia: Phalobase: Trab long, thick, distally curved, Mulsant. and expanded. Basal piece rectangular. Parameres short, Host: It is a general predator of aphids, scale insects, comparatively thin, tips pointed and with long hairs. and mites. Median lobe thick, broad at base, gradually tapering Habitat: It can be found on the lower sides of fruit trees toward apex, tip rounded with long hairs on dorsal and like apple and walnut, and on grape vines. distal portions. Siphon: Capsule unique, asymmetrical, Local distribution: Jaglot, Gilgit. both arms forming 180° angle, outer arm short, inner arm World distribution: Bangladesh, China, India, long. Tube cylindrical, moderate in thickness, abruptly Myanmar, Nepal, Pakistan, Sri Lanka. curved at base, then broadly curved at middle, distal 3.4.2. Scymnus posticalis Sicard, 1913 portion comparatively thin, tip flattened with a narrow Synonyms: Scymnus posticalis Sicard, 1913; Scymnus process directed downward. Male genitalia of Scymnus (Pullus) posticalis Korschefsky, 1931; –Pong, 1984. Scymnus posticalis shown in Figure 5. hilaris Weise, 1879; –Lewis, 1896. Pullus hilaris Ohta, Remarks: This relatively small black coccinellid species 1929. Pullus hilaris ab. awanus Ohta, 1929; –Sasaji, 1971. resembles other members of genus Scymnus like S. nubilus Scymnus (Pullus) hilaris Sasaji, 1971. Scymnus (Scymnus) and S. pauperculus, as all 3 species are similar in size and ishidai Araki, 1963; –Sasaji, 1971. Scymnus (Nephus) inops body coloration. However, this species is remarkably Smirnoff, 1973. different in the male genitalia structure from the other 2 Material examined: 1 ♂ 2 ♀ Pak. GB, Hunza-Nager, species. Thapa (2000) reported this species from Nepal. Gulmit, 04.VI.2007 (M. Ashfaque) (NIM); 2 ♂ 3 ♀ Pak. Agarwala and Yasuda (2001) reported this species as a GB, Hunza-Nager, Gulmit, 12.VI.2008 (Ashraf) (NIM). predator of Aphis gossypii from India. Poorani (2002) Diagnosis: Body small, oval, highly convex, ground also listed S. posticalis in the Indian checklist of predatory color black and densely pubescent (Figure 4). Head coccinellids. brownish black, not visible from above, eyes small, Host: It is a general predator of aphids, scale insects, brownish black. Pronotum black, densely pubescent, and mites. transverse, curved inward, posteriorly pushing scutellum. Habitat: It can be found on fruit trees like apple and Scutellum visible and black. Elytra shiny black except anal walnut, and on grapes. portion, which is light reddish brown. Local distribution: Gulmit, Hunza.

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Figure 5. Male genitalia of Scymnus posticalis.

World distribution: China, India, Japan, Myanmar, Figure 6. Adult of Stethorus gilvifrons. Nepal. 3.5. Genus: Stethorus Weise, 1885 3.5.1. Stethorus gilvifrons (Mulstant, 1850) Synonyms: Scymnus gilvifrons Mulsant, 1850; –Crotch, 1874; –Korschefsky, 1931; –Kapur, 1948; –Mathur and Srivastava, 1966. Scymnus (Stethorus) gilvifrons Weise, 1885. Material examined: 3 ♂ 4 ♀ Pak. GB, Gilgit, Jaglot, 03.VII.2003 (Falak Naz) (NIM); 2 ♂ 2 ♀ Pak. GB, Gilgit, 04.VI.2007 (M. Ashfaque) (NIM); 3 ♀ Pak. GB, Gilgit, Jaglot, 15.VI.2008 (Amjad) (NIM). Diagnosis: Body small, slightly oval, widest at middle, convex, black, coarsely punctate, pubescent with rather long white hairs as shown in Figure 6. Head mostly black with anterior half testaceous and sparsely punctate. Pronotum and elytra black. Dorsal surface with grayish, moderately long, sparse, semierect hairs. Legs testaceous. Male genitalia with thin parameres, shorter than median lobe, median lobe very long, curved at subdistal portion, tip of siphonal tube blunt. Male genitalia: Phalobase: Trab very short and distally expanded. Basal piece: Small and semicircular. Median Figure 7. Male genitalia of Stethorus gilvifrons. lobe: Very long, cylindrical, narrow and tapering toward apex, bent toward parameres at subdistal portion slightly and without hairs. Parameres: Very thin, about three- Remarks: The type locality of this species is Derbent, quarters of median lobe without hairs, apex with a pair of Caucasus (USSR). Kapur (1948), while working on the setae. Siphon: Siphonal capsule simple, without an outer genus Stethorus Weise of Old World species, described this hook but inner hook elongated and apically bifurcated, species. He also reported some materials of this species siphonal tube thin, long, forming a complete loop with from Mastung (Balochistan) and Faisalabad (formerly pointed tip as shown in Figure 7. Lyallpur) (Punjab), Pakistan, collected while feeding on

1038 ASHFAQUE et al. / Turk J Zool mites of apple and castor oil plants. It was also reported by Rafi et al. in 2005 from northern Pakistan without consulting genitalia. This species may be confused with another Stethorus species, S. pauperculus. Both species are very similar in general appearance and coloration. However, it can be distinguished from S. pauperculus by the structure of the male genitalia. The siphon of S. gilvifrons forms a complete loop, while that of S. pauperculus is somewhat straight. Some authors have incorrectly illustrated the male genitalia of this species with S. siphonulus. This species is also confused with S. wollastoni Kapur (Syn. S. minimus) due to similarities in coloration, especially the testaceous interior part of head. Host: It is a general predator of mites. Habitat: It can be found in fruit orchards and on isolated fruit trees like pear, apple, and walnut, and on grapes. Local distribution: Jaglot, Gilgit. World distribution: Cyprus, India, Iran, Italy, Pakistan, Saudi Arabia, Yemen. 3.5.2. Stethorus pauperculus (Weise, 1895) Synonyms: Scymnus (Stethorus) pauperculus Weise, 1895; Figure 8. Adult of Stethorus pauperculus. Stethorus pauperculus Weise, 1900; –Kapur, 1948. Material examined: 2 ♂, 2 ♀ Pak. GB, Gilgit, 03.VI.2003 (Ather) (NIM); 1 ♂ 5 ♀ Pak. GB, Gilgit, Jaglot, 03.VII.2003 (Falak Naz) (NIM); 3 ♂ 1 ♀ Pak. GB, Gilgit, 04.VI.2007 (M. Ashfaque) (NIM). Diagnosis: Body small with yellow mouth parts. Elytra and thorax wholly black, devoid of spots, pubescence distinctly apparent, legs brownish, short, and profusely setose. Adult is shown in Figure 8. Genitalia: Phalobase: Trab with short apex slightly expanded. Basal piece: Semicircular. Parameres: Long, slender, gradually tapering toward apex forming hairs like tip. Median lobe: Very long cylindrical thickness up to three-quarters of its length, then slightly turned and tapering toward apex. Siphonal tip: abruptly turned and pointed. Siphon capsule absent, small tube replacing capsule. Siphonal tube turned abruptly, slightly bent at middle, tip blunt. Male genitalia are given in Figure 9. Remarks: This species also resembles other Stethorus species. Its distinguishing characters from S. gilvifrons are given under the remarks for S. gilvifrons. This species can be distinguished from S. yunnanensis by comparing male genitalia. Siphonal tube in S. pauperculus is somewhat straight at the base, while that of S. yunnanensis is semicircular at the base. Median lobe of S. pauperculus is comparatively thin with rounded tip, whereas it is broad Figure 9. Male genitalia of Stethorus pauperculus. with notched tip in S. gilvifrons. Host: It is a general predator of mites. Habitat: It can be found in fruit orchards and on Local distribution: Jaglot, Gilgit. isolated fruit trees like pear, apple, and walnut, and on World distribution: India, Malaysia, Pakistan, grapes. Thailand, Yemen.

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4. Discussion and S. pauperculus. Among these, Scymnus posticalis is a Previously, Irshad (2001, 2003) and Rafi et al. (2005) new record for the Gilgit-Baltistan region, a high-altitude reported 3 species of subfamily Scymninae from the dry mountain area of Pakistan. The taxonomic position Northern Areas of Pakistan (now Gilgit-Baltistan). These of these species was confirmed by genitalia morphology; include Scymnus nubilus Mulstant, 1850; Stethorus gilvifrons previously, identification was carried out only through (Mulstant, 1850); and Stethorus pauperculus (Weise, external morphology of the adults, which may result in 1895). During the present study, surveys were conducted misidentification. Micrococcinellids, especially species of throughout the Gilgit-Baltistan region for ladybird beetles genera Scymnus and Stethorus, are similar in appearance of the subfamily Scymninae. The taxonomic study of and need to be identified through genitalia examination. the collected specimens confirmed the occurrence of 4 The findings are significant, with 4 confirmed records of species in 2 genera of subfamily Scymninae in the region. Scymninae, and could serve as a baseline study for the The identified species occurring in Gilgit-Baltistan are region of Gilgit-Baltistan. Scymnus nubilus Mulstant, S. posticalis, Stethorus gilvifrons,

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