Description of Sandfly (Diptera: Psychodidae: Phlebotominae) Species from Genus  Sergentomyia (Franca and Parrot, 1920) in Sri Lanka

Gajapathy, K.; Surendran, S.N.

ABSTRACT

Leishmaniasis, mainly Cutaneous Leishmaniasis (CL), is a major public health concern in Sri Lanka. Although, the vector for any form of the disease is yet to be clearly discriminated in the country, a limited study reported the potential role of Phlebotomus argentipes s.l. in the transmission of Leishmania donovani, the causative agent of CL in Sri Lanka. In a survey from three Cutaneous Leishmaniasis (CL) endemic localities in Sri Lanka, a rich diversity of the genus Sergentomyia was discovered. Eleven species belong to three subgenera namely, Parrotomyia, Neophlebotomus and Grassomyia were identified and their detail morphology is described in this report. Only three species of Parrotomyia and one from each Neophlebotomus and Grassomyia have been reported previously from Sri Lanka. Eight species of Parrotomyia, namely Se. (Par.) rudnicki, Se. (Par.) babu insularis, Se. (Par.) modii, Se. (Par.) baghdadis, Se. (Par.) barraudi, Se. (Par.) grekovi, Se. (Par.) bailyi and Se. (Par.) species were recorded during the survey. Sergentomyia (Neophlebotomus) malayae, Sergentomyia (Grassomyia) dreyfussi turkestanica and Se. (Gra.) indica belong to the subgenera Neophlebotomus and Grassomyia were also recorded during this study. This is the first record for Sergentomyia (Neophlebotomus) malayae and Sergentomyia (Grassomyia) dreyfussi turkestanica and for all the species of Parrotomyia except Se. (Par.) babu insularis and Se. (Par.) barraudi. The study warrants for an extensive sandfly survey, especially in endemic areas, as Sergentomyia species are now considered as potential vectors for many parasites including Leishmania.

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INTRODUCTION

Sandflies are vectors of many parasites including arboviruses, bartonelloses and more specifically Leishmania (Protozoa: Zoomastigophora: Kinetoplastida: Trypanosomatidae) (Ashford, 2000; Alexander and Maroli, 2003). The common diseases caused by them are sandfly fever (Tavana, 2007) and leishmaniasis. Leishmania sp. causes various forms of leishmaniasis such as Cutaneous Leishmaniasis (CL), Mucocutaneous Leishmaniasis (MCL) and Visceral Leishmaniasis (VL). More than 80 countries are affected by the disease (Alexander and Maroli, 2003).

Leishmaniasis emerges as a major public health threat in Sri Lanka. The first autochthonous case of CL was reported in 1992 (Athukorale et al., 1992). Several cases have been reported from different parts of the country since then. More than 2000 CL cases were reported in the country until 2009 (Siriwardana et al., 2010). The first case of Visceral Leishmaniasis (VL) was identified from North Central Province in 2006 (Abeygunasekara et al., 2007).

Thirty out of 700 recorded sandfly species are reported to be vectors of leishmaniasis (Ilango, 2011). Many species from the genus Phlebotomus Rondani and Berte in Rondani is generally considered as vector for VL and CL. Members from the genus Sergentomyia Franca and Parrot. 1920 have not drawn much attention as they were generally regarded to feed on reptiles and small mammals (Artemiev, 1978; Lewis, 1978). However, some findings suggest their potential role in the transmission of leishmaniasis in the oriental region (Mutinga, 1986; Maroli et al., 1988; Mukherjee et al., 1997; Geevarghese et al., 2004). Sergentomyia consists of seven subgenera including Parrotomyia (Theodor, 1948), Neophlebotomus (Franca and Parrot, 1920) and Grassomyia (Theodor, 1948). Several species have been described under the subgenus Parrotomyia from the oriental region. Grassomyia consists of two species and few subspecies, while Neophlebotomus consists of as many as 60 species (Lewis, 1978).

The taxonomic studies on Sri Lankan sandflies are fragmentary. Most of the records are dated back to the mid twentieth century (Carter and Antonipulle, 1949; Lewis, 1978) except a few recent studies (Surendran et al., 2005a, b; Ozbel et al., 2011; Gajapathy and Surendran, 2011; Gajapathy et al., 2011). CL is considered as a notifiable disease in Sri Lanka. Proper identification of sandfly species and studying their distribution pattern are essential in formulating an appropriate vector control programme. In addition, cataloging sandfly species in the country is essential to identify potential vectors of CL and VL.

In this background, a survey was conducted in few CL endemic localities in Sri Lana to catalogue Sergentomyia species. The current study reports the presence of subgenera Parrotomyia, Grassomyia and Neophlebotomus.

MATERIALS AND METHODS

Sampling was done in CL endemic localities in the districts of Vavuniya (8°45' 0" N/80°30' 0" E), Trincomalee (8°34' 0" N/81°14' 0" E) and Anuradhapura (8°21' 0" N/80°23' 0" E) in Sri Lanka (Fig. 1). All these areas are in the dry zone of the country.


Fig. 1:	Sandfly collection sites, Pa: Padaviya, P: Padavi Siripura, V: Vavuniya

Sandflies were collected in and around the houses and warehouses in the areas by a mouth aspirator form 1700 to 0500 h from August 2010 to April 2011. The flies were preserved in 70% ethyl alcohol. Identification was done at the Zoology research laboratory of the University of Jaffna. Each fly was processed and dissected following the techniques described by Lewis (1978), Young and Duncan (1994), Aslamkhan and Aslamkhan (2000), Kakarsulemankhel (2004a) and Singh et al. (2007).

Microscopic analysis was done with Kyowa model SE-L (Japan), Olympus model BX51 (Japan) and a monocular light microscope (Kyowa, Japan) equipped with a moving vernier scale and an ocular micrometer. Permanent slides were prepared with Canada balsam (Ko et al., 2008). Drawings were done in the Camera Lucida attached to BX51 (Olympus, Japan).

Identification of specimens was carried out with the available taxonomic keys (Lewis, 1978; Lane, 1993; Kakarsulemankhel, 2004b-e, 2008). Prepared permanent slides with all the collection details were deposited in the Department of Zoology, University of Jaffna.

RESULTS

Subgenus: Parrotomyia: Thirty eight individual flies, belong to the genus Sergentomyia and the subgenus Parrotomyia, were collected. The presence of morphological characters like pointed wing, the small oval hair sockets in the second and subsequent abdominal tergites and four style spines (males), longer third antennal segment (Aiii) (>labrum) and smooth elliptical to round spermatheca (female) were observed in flies and R₂/R₂₊₃ (wing index value) was less than 1.0.

Identified species
Sergentomyia (Parrotomyia) rudnicki (Lewis, 1978)
Specimen examined
Female: Not found in the collection.

Male (4 specimens): Abdomen length 0.9 mm; abdomen breadth 0.32 mm; thorax length 0.48-0.49 mm; thorax breadth 0.45 mm; head length 0.31-0.32 mm; head breadth 0.22 mm; eye length 0.14 mm; eye breadth 0.09-0.1 mm; inter ocular 0.09 mm; wing length 1.65-1.66 mm; wing breadth 0.33 mm; halter length 0.14 mm. fore-femur: fore-tibia: fore-basi tarsi 4:3.1:1.6; mid-femur: mid-tibia: mid-basi tarsi 3.9:3.5:1.8; hind-femur: hind-tibia: hind-basi tarsi 4:4.0:2.1.

Mouth parts: Labrum is chitinized stout structure with three apical sensilla. The hypopharynx is narrow. Maxillary blade has very fine teeth scattered. Cibarium (Fig. 2b) is with a triangle shaped pigment patch with a very small forward process. 25-26 teeth are found. Pharynx is slender without large constriction. Armature is found but it is faint (Fig. 2c).

Labrum length 0.1 mm; maxillary palp length 0.93 mm; palp formula (1, 2, 3, 4, 5) with a ratio of 7:5:13:18.5:22 proboscis length 0.15 mm; cibarial length 0.4 mm; third antennal segment (Aiii) 0.21-0.22 mm; Aiv 0.11 mm; Av 0.11 mm; antennal ascoid sensilla chaetica on third antennal segment (AaIII) 0.03 mm; Aaiv 0.03 mm; Aav 0.03 mm. Ascoid formula is 1/3-15.

Wing venation: First radial vein (R₁) 0.68 mm; R₂ 0.17 mm; R₃ 0.27-0.28 mm; R₂₊₃ 0.28 mm; R₁ overlap 0.07 mm.

Genitalia: Coxa is slightly broader at middle; style has four spines with equal length; two at terminal and two at sub terminal with the accessory setae in middle. Style is 5 times longer than the breadth.


Fig. 2(a-c):	Male Sergentomyia (Parrotomyia) rudnicki Lewis showing: (a) Terminalia, Pm: Paramere, SS: Surstyle, As: Aedegal sheath, (b) Cibarium, CT: Cibarial teeth and (c) Pharynx, PA: Pharyngeal armature

Paramere is slender and hooked (Fig. 2a). Aedegal sheath is slender tapering downwards with round end (Fig. 2a).

Coxa length 0.25-0.26 mm; coxa breadth 0.04 mm; style length 0.11 mm; style breadth 0.02 mm; paramere lobe length 0.14 mm; aedegal sheath length 0.08 mm; genital pump length 0.08 mm; genital filament length 0.26 mm.

Sergentomyia (Parrotomyia) baghdadis (Adler and Theodor 1929)
Specimen examined
Male: Not found in the collection.

Female (4 specimens): Abdomen length 0.69-0.70 mm; abdomen breadth 0.24-0.26 mm; thorax length 0.45 mm; thorax breadth 0.39-0.40 mm; head length 0.27-0.28 mm; head breadth 0.18-0.20 mm; eye length 0.17 mm; eye breadth 0.15 mm; inter ocular 0.11 mm; wing length 1.40-1.41 mm; wing breadth 0.34-0.35 mm; halter length 0.18 mm. fore-femur: fore-tibia: fore-basi tarsi 6.5:19:15; mid-femur: mid-tibia: mid-basi tarsi 7:19:16; hind-femur: hind-tibia: hind-basi tarsi 8:18:20.

Mouth parts: Labrum is strongly chitinized pointed structure with three apical sensilla. The hypopharynx is broader compared to that of male with a marginal minute serration. Mandible is with serrated teeth at the edge. Maxillary blade has visible teeth. Cibarium (Fig. 3d) is with 20 teeth in a convex plate with a light brown pigment patch with a notch at hind-plate. Pharynx is slender and with scales and minute teeth at base (Fig. 3c).


Fig. 3(a-d):	Female Sergentomyia (Parrotomyia) baghdadis Adler and Theodor showing: (a) Fourth antennal segment, Aa: Antennal ascoid, (b) Spermatheca (Sp), (c) Pharynx, PA: Pharyngeal armature and (d) Cibarium, CT: Cibarial teeth

Labrum length 0.12 mm; maxillary palp length 0.5 mm; palp formula (1, 2, 3, 4, 5) with a ratio of 3.5:4.5:6.5:7:8.5; proboscis length 0.14 mm; cibarial length 0.04 mm; third antennal segment (Aiii) 0.20-0.21 mm (Fig. 3a); Aiv 0.07 mm; Av 0.07-0.08 mm; antennal ascoid/sensilla chaetica on third antennal segment (Aaiii) 0.02 mm; Aaiv 0.02 mm; Aav 0.02-0.03 mm.

Wing venation: First radial vein (R₁) 0.51-0.52 mm; R₂ 0.22 mm; R₃ 0.3-0.33 mm; R₂₊₃ 0.24 mm; R₁ overlap 0.10 mm.

Genitalia: Elliptical spermatheca (Fig. 3b) with bulged middle and a smooth wall. Tip is with minute projections. The individual ducts are fused to form a short common duct and then fall in to the genital atrium. Spermatheca length 0.03 mm; breadth 0.02 mm.

Sergentomyia (Parrotomyia) species (Kaul et al., 1973): The species was described from Kota district, India. Lewis (1978) stated that this could be related to Se. (Par.) grekovi.

Specimen examined
Female: Not found in the collection.

Male (10 specimens): Abdomen length 0.66-0.68 mm; abdomen breadth 0..33-0.34 mm; thorax length 0.46-0.50 mm; thorax breadth 0.36-0.37 mm; head length 0.29-0.32 mm; head breadth 0.22-0.23 mm; eye length 0.15 mm; eye breadth 0.11 mm; inter ocular 0.10 mm; wing length 1.15-1.16 mm; wing breadth 0.24 mm; halter length 0.19 mm. Fore-femur: fore-tibia: fore-basi tarsi 13:10:6; mid-femur: mid-tibia: mid-basi tarsi 13.5:13:7; hind-femur: hind-tibia: hind-basi tarsi 15:17:9.5.

Mouth parts: Labrum is chitinized blunt, broad structure. The hypopharynx is narrow with bristle like serration (soft). Maxillary blade has very fine teeth (2-3). Cibarium (Fig. 4a) is with a dull pigment patch. Convex arrangement of teeth is with larger (0.002 mm) laterals (5-6) and smaller minute central teeth (0.001 mm). Pharynx is broader at base without constriction. Pharynx armature is with scales and ridges.

Labrum length 0.11 mm; maxillary palp length 0.48-0.50 mm; palp formula (1, 2, 3, 4, 5) with a ratio of 2:5:8:8.5:17; proboscis length 0.156 mm; cibarial length 0.024 mm; third antennal segment (Aiii) 0.08 mm (Fig. 4b); Aiv 0.05-0.06 mm; Av 0.06 mm; antennal ascoid/sensilla chaetica on third antennal segment (Aaiii) 0.03 mm; Aaiv 0.02 mm; Aav 0.02-mm; ascoid formula is 1/3-15.

Wing venation: First radial vein (R₁) 0.45-0.47 mm; R₂ 0.12 mm; R₃ 0.24-0.26 mm; R₂₊₃ 0.16-0.17 mm; R₁overlap is not found.

Genitalia: Coxa is broad from base and tapper; style (Fig. 4d) has four spines with equal length, with two sub-apical and two at apex with accessory setae in middle. Paramere is hooked. Aedegal sheath (Fig. 4c) is thick and slightly curved downwards.

Coxa length 0.18 mm; coxa breadth 0.05 mm; style length 0.06 mm; style breadth 0.02 mm; paramere lobe length 0.14 mm; aedegal sheath length 0.12 mm; genital pump length 0.07 mm; genital filament length 0.23 mm.

Sergentomyia (Parrotomyia) barraudi (Sinton, 1929)
Specimen examined
Female (4 specimens): Abdomen length 0. 80 mm; abdomen breadth 0.35 mm; thorax length 0.48 mm; thorax breadth 0.48 mm; head length 0.24 mm; head breadth 0.18 mm; eye length 0.15 mm; eye breadth 0.10 mm; inter ocular 0.12 mm; wing length 1.3 mm; wing breadth 0.33 mm; halter length 0.54 mm.


Fig. 4(a-d):	Male Sergentomyia (Parrotomyia) species A Kaul, Dhanda and Modi showing: (a) Cibarium, CT: Cibarial teeth, (b) Fourth antennal segment, Aa: Antennal ascoid, (c) Aedegal sheath (AS) and (d) Style, SS: Style spines, Set: Setae


Fig. 5(a-c):	Male Sergentomyia (Parrotomyia) barraudi Sinton showing: (a) Male terminalia, Pm.l: Paramere lobe, (b) Style, S.S.: Style spines and (c) Fourth antennal segment, Aa: Antennal ascoid

Mouth parts: Labrum is strongly chitinized. The hypopharynx is broader compared to that of male with a marginal soft serration. Mandible is broad with serrated teeth found along the edge. Cibarium is with 40 teeth in a straight plate. The pigment patch is bifid. Pharynx is slender. Pointed teeth are found in the armature with a convex at posterior.

Labrum length 0.05 mm; maxillary palp length 0.45 mm; palp formula (1, 2, 3, 4, 5) with a ratio of 12:20:32:38:78; proboscis length 0.07 mm; cibarial length 0.04 mm; third antennal segment (Aiii) 0.12 mm (Fig. 5b); Aiv 0.06 mm; Av 0.05 mm; antennal ascoid/sensilla chaetica on third antennal segment (Aaiii) 0.012 mm; Aaiv 0.01 mm; Aav 0.01 mm.

Wing venation: First radial vein (R₁) 0.55 mm; R₂ 0.14 mm; R₃ 0.24 mm; R₂₊₃ 0.21 mm; R₁ overlap 0.02 mm.

Genitalia: Elliptical spermatheca with smooth wall. Spermatheca length 0.06 mm; breadth 0.02.

Male (4 specimens): Abdomen length 0.75 mm; abdomen breadth 0.27-0.28 mm; thorax length 0.36 mm; thorax breadth 0.39 mm; head length 0.27-0.28 mm; head breadth 0.2 mm; eye length 0.14 mm; eye breadth 0.08 mm; inter ocular 0.1 mm; wing length 1.18-1.19 mm; wing breadth 0.26 mm; halter length 0.18 mm. fore-femur: fore-tibia: fore-basi tarsi 15:12:8; mid-femur: mid-tibia: mid-basi tarsi 15.5.14:9; hind-femur: hind-tibia: hind-basi tarsi 16:17:10.

Mouth parts: Labrum is chitinized broad structure. The hypopharynx is narrow with soft bristle like serration. Maxillary blade has very fine teeth. Cibarium is with a light, wider pigment patch. Convex arrangement of 18 teeth is found. Pharynx is slender. Pharynx armature is with very faint ridge like armature.

Labrum length 0.13 mm; maxillary palp length 0.53-0.54 mm; palp formula (1,2,3,4,5) with a ratio of 76:40:31:23:09; proboscis length 0.15 mm; cibarial length 0.02 mm; third antennal segment (Aiii) 0.15-0.16 mm; Aiv 0.08 mm; Av 0.08 mm; antennal ascoid/sensilla chaetica on third antennal segment (Aaiii) 0.02 mm; Aaiv 0.02 mm; Aav 0.02 mm; ascoid formula is 1/3-15.

Wing venation: First radial vein (R₁) 0.51 mm; R₂ 0.18 mm; R₃ 0.23-0.24 mm; R₂₊₃ 0.17 mm; R₁overlap 0.06 mm.

Genitalia: Coxa is longer with slightly bulged middle (Fig. 5a). Style has four spines with equal length, with two sub-apical and two at apex with accessory setae in middle proximal to the sub-apical spines. Paramere is thin and hooked. Aedegal sheath is slender and curved downwards. Coxa length 0.20 mm; coxa breadth 0.06 mm; style length 0.14 mm; style breadth 0.04 mm; paramere lobe length 0.15-0.16 mm; aedegal sheath length 0.07 mm; genital pump length 0.08 mm; genital filament length 0.24-0.25 mm.

Sergentomyia (Parrotomyia) modii (Lewis, 1978)
Specimen examined
Male: Not found in the collection.

Female (6 specimens): Abdomen length 0. 84-0.85 mm; abdomen breadth 0.36 mm; thorax length 0.46-0.50 mm; thorax breadth 0.41-0.42 mm; head length 0.33 mm; head breadth 0.295 mm; eye length 0.13-0.14 mm; eye breadth 0.11 mm; inter ocular 0.10 mm; wing length 1.29 mm; wing breadth 0.39 mm; halter length 0.54 mm. fore-femur: fore-tibia: fore-basi tarsi 4:4:2; mid-femur: mid-tibia: mid-basi tarsi 4:4.2:2.1; hind-femur: hind-tibia: hind-basi tarsi 4.1.4.5:2.3.

Mouth parts: Labrum is chitinized with 2 apical sensilla and 2 laterals at each side. The hypopharynx (Fig. 6b) is broad with 6 marginal teeth. Mandible is broad with serrated teeth found along the edge. Maxillary blade has visible teeth (16 lateral and 10 ventral). Cibarium is with broad teeth in a straight plate. The pigment patch is cone shaped. Pharynx is slender with scale like armature.

Labrum length 0.15 mm; maxillary palp length 0.60 mm; palp formula is 1,2,(3,4),5 with a ratio of 92:40:31:31:8; proboscis length 0.17 mm; cibarial length 0.36 mm; third antennal segment (Aiii) 0.12-0.13 mm (Fig. 6a); Aiv 0.6-0.66 mm; Av 0.66 mm; antennal ascoid/sensilla chaetica on third antennal segment (Aaiii) 0.02 mm; Aaiv 0.02 mm; Aav 0.02 mm.


Fig. 6(a-c):	Female Sergentomyia (Parrotomyia) modii Lewis showing: (a) Fourth antennal segment, Aa: Antennal ascoid, (b) Hypopharynx (HP) and (c) Spermatheca (Sp), ID: Individual duct

Wing venation: First radial vein (R₁) 0.58-0.59 mm; R₂ 0.22 mm; R₃ 0.30 mm; R₂₊₃ 0.24 mm; R₁ overlap 0.1 mm.

Genitalia: Elliptical spermatheca (Fig. 6c) with smooth wall. Spermatheca length 0.04 mm; breadth 0.02 mm. The length of cerci 0.12 mm; length of surstyle 0.06 mm.

Sergentomyia (Parrotomyia) babu insularis (Theodor, 1938)
Specimen examined
Female: Not found in the collection.

Male (1 specimen): Abdomen length 0.9 mm; abdomen breadth 0.27 mm; thorax length 0.42 mm; thorax breadth 0.45 mm; head length 0.32 mm; head breadth 0.27 mm; eye length 0.15 mm; eye breadth 0.11 mm; inter ocular 0.1 mm; wing length 1.29 mm; wing breadth 0.33 mm; halter length 0.57 mm. fore-femur: fore-tibia: fore-basi tarsi 18:18:9; mid-femur: mid-tibia: mid-basi tarsi 19:21:9; hind-femur: hind-tibia: hind-basi tarsi 19:25:11.

Mouth parts: Labrum is chitinized structure with 3 apical sensilla. The hypopharynx is narrow with 16 soft bristle like serration. Maxillary blade has very fine teeth. Cibarium (Fig. 7b) is with a light and wider pigment patch. Concave arrangement of teeth is found; 40 in number. Pharynx is slender. Pharynx armature is with scales.

Labrum length 0.15 mm; maxillary palp length 0.66 mm; palp formula (1,2,3,4,5) with a ratio of 90:46:36:32:13; proboscis length 0.17 mm; cibarial length 0.02 mm; third antennal segment (Aiii) 0.19 mm; Aiv 0.1 mm; Av 0.99 mm; antennal ascoid/sensilla chaetica on third antennal segment (Aaiii) 0.02 mm; Aaiv 0.02 mm; Aav 0.02 mm; ascoid formula is 1/3-15.

Wing venation: First radial vein (R₁) 0.98 mm; R₂ 0.22 mm; R₃ 0.34 mm; R₂₊₃ 0.22 mm; R₁overlap 0.01 mm.

Genitalia: Coxa is longer with slightly bulged middle (Fig. 7a). Style has four spines with equal length; with two sub-apical and two at apex with accessory setae in middle proximal to the sub-apical spines. Paramere is thin and slightly hooked. Aedegal sheath is slender.

Coxa length 0.25 mm; coxa breadth 0.07 mm; style length 0.11 mm; style breadth 0.02 mm; paramere lobe length 0.15 mm; aedegal sheath length 0.09 mm; genital pump length 0.09 mm; genital filament length 0.32 mm.


Fig. 7(a-b):	Male Sergentomyia (Parrotomyia) babu insularis Theodor showing (a) Male terminalia, St: Style, Set: Setae, S.S.: Style spines, Pm.l.: Paramere and Sst: Surstyle and (b) Cibarium, CT: Cibarial teeth

Sergentomyia (Parrotomyia) grekovi (Khodukin, 1929)
Specimen examined
Male: Not found in the collection.

Female (1 specimen): Abdomen length 0.84-0.85 mm; abdomen breadth 0.36 mm; thorax length 0.46-0.50 mm; thorax breadth 0.41-0.42 mm; head length 0.33 mm; head breadth 0.3 mm; eye length 0.13-0.14 mm; eye breadth 0.1 mm; inter ocular 0.1 mm; wing length 1.29 mm; wing breadth 0.39 mm; halter length 0.54 mm. fore-femur: fore-tibia: fore-basi tarsi 4:4:2; mid-femur: mid-tibia: mid-basi tarsi 4:4.2:2.1; hind-femur: hind-tibia: hind-basi tarsi 4.1.4.5:2.3.

Mouth parts: Labrum is chitinized with 2 apical sensilla and 2 laterals at each side. The hypopharynx is broad with 6 marginal teeth. Mandible is broad with serrated teeth found along the edge. Maxillary blade has visible teeth (16 lateral and 10 ventral). Cibarium (Fig. 8b) is with broad teeth in a straight plate. The pigment patch is cone shaped. Pharynx is slender with scale like armature (Fig. 8a).

Labrum length 0.15 mm; maxillary palp length 0.60 mm; palp formula is 1,2,(3,4),5 with a ratio of 92:40:31:31:8; proboscis length 0.17 mm; cibarial length 0.36 mm; third antennal segment (Aiii) 0.12-0.13 mm; Aiv 0.6-0.66 mm; Av 0.66 mm; antennal ascoid/sensilla chaetica on third antennal segment (Aaiii) 0.02 mm; Aaiv 0.02 mm; Aav 0.02 mm.

Wing venation: First radial vein (R₁) 0.58-0.59 mm; R₂ 0.22 mm; R₃ 0.30 mm; R₂₊₃ 0.24 mm; R₁overlap 0.09-0.1 mm.

Genitalia: Elliptical spermatheca with smooth wall. Spermatheca length 0.04 mm; breadth 0.02 mm. The length of cerci 0.12 mm; length of surstyle 0.06 mm.

Sergentomyia (Parrotomyia) bailyi (Sinton, 1931)
Specimen examined
Male: Not found in the collection.


Fig. 8(a-b):	Female Sergentomyia (Parrotomyia) grekovi Khodukin showing: (a) Pharynx PA: Pharyngeal armature and (b) Cibarium, CT: Cibarial teeth


Fig. 9:	Cibarium of female Sergentomyia (Parrotomyia) bailyi Sinton, CT: Cibarial teeth

Female (4 specimens): Abdomen length 0. 84-0.85 mm; abdomen breadth 0.45-0.46 mm; thorax length 0.45-0.46 mm; thorax breadth 0.41-0.42 mm; head length 0.27-0.28 mm; head breadth 0.25-0.26 mm; eye length 0.13-0.14 mm; eye breadth 0.1-0.12 mm; inter ocular 0.10 mm; wing length 1.35 mm; wing breadth 0.33-0.35 mm; halter length 0.57-0.59 mm. fore-femur: fore-tibia: fore-basi tarsi 14:16:8; mid-femur: mid-tibia: mid-basi tarsi 15:18:8.5; hind-femur: hind-tibia: hind-basi tarsi 15.5:20:10.

Mouth parts: Labrum is chitinized with 3 apical sensilla. The hypopharynx is broad with marginal soft serration. Cibarium (Fig. 9) is with small pointed teeth at slightly convex plate. The pigment patch is dark and overlapping teeth. Pharynx is slender with scale like prominent armature.

Labrum length 0.14-0.15 mm; maxillary palp length 0.51-0.52 mm; palp formula is 1,2,3,4,5 with a ratio of 15:10.5:8:6:3.5; proboscis length 0.16-0.166 mm; cibarial length 0.26 mm; third antennal segment (Aiii) 0.11-0.12 mm; Aiv 0.59-0.6 mm; Av 0.62 mm; antennal ascoid/sensilla chaetica on third antennal segment (Aaiii) 0.02 mm; Aaiv 0.02 mm; Aav 0.02 mm.

Wing venation: First radial vein (R₁) 0.55 mm; R₂ 0.22 mm; R₃ 0.30-0.31 mm; R₂₊₃ 0.22-0.23 mm; R₁ overlap 0.09-0.1 mm.

Genitalia: Elliptical spermatheca with striated wall. The ducts are striated. Spermatheca length 0.09 mm; breadth 0.06 mm. The length of cerci 0.13 mm; length of surstyle 0.05 mm.

Subgenera: Grassomyia and Neophlebotomus: Ten flies belonging to the sub genera Grassomyia and Neophlebotomus from the genus Sergentomyia were collected.

Identified species
Sergentomyia (Grassomyia) dreyfussi turkestanica (Theodor and Mesghali, 1964)
Specimen examined
Male: Not found in the sample.

Female: 4 specimens: Abdomen length 1.5. mm; abdomen breadth 0.520-0.540 mm; thorax length 0.460-0.480 mm; thorax breadth 0.510 mm; head length 0.300 mm; head breadth 0.200 mm; eye length 0.130 mm; eye breadth 0.100 mm; inter ocular 0.110 mm; wing length 0.140-0.150 mm; wing breadth 0.360 mm; halter length 0.140 mm. fore-femur: fore-tibia: fore-basi tarsi 17:13:7; mid-femur: mid-tibia: mid-basi tarsi 17:17:11; hind-femur: hind-tibia: hind-basi tarsi 21:25:11. First femora are with spines.

Mouth parts: Labrum is strongly chitinized. Mandible is with serrated teeth at the edge. Maxillary blade has visible teeth (18 laterals and 6 ventrals). Cibarium (Fig. 10a) is with 40 long pointed teeth in a convex plate with a brown pigment patch. Pharynx (Fig. 10b) is bottle shaped and with minute teeth at base (Fig. 10b).

Labrum length 140 mm; maxillary palp length 0.540 mm; palp formula (1,2,3,4,5) with a ratio of 10:15:32:41:45; proboscis length 0.150 mm; cibarial length 0.400 mm; third antennal segment (Aiii) 0.120 mm; Aiv 0.070 mm; Av 0.070-0.075 mm; antennal ascoid/sensilla chaetica on third antennal segment (Aaiii) 0.033 mm; Aaiv 0.035 mm; Aav 0.035 mm.

Wing venation: First radial vein (R₁) 0.700 mm; R₂ 0.210-0.220 mm; R₃ 0.320-0.325 mm; R₂₊₃ 0.240 mm; R₁ overlap 0.030 mm.

Genitalia: Round small spermatheca (Fig. 10c) with a tip with small projections. The individual ducts are fused to form a short common duct and then fall in to the genital atrium. Spermatheca length 0.024 mm; breadth 0.030 mm.

Sergentomyia (Grassomyia) indica (Theodor); (Theodor and Mesghali, 1964)
Specimen examined
Male: Not found in the collection.

Female: (2 specimens): Abdomen length 0.900 mm; abdomen breadth 0.380 mm; thorax length 0.440 mm; thorax breadth 0.420 mm; head length 0.420 mm; head breadth 0.170 mm; eye length 0.120 mm; eye breadth 0.120 mm; inter ocular 0.100 mm; wing length 0.125 mm; wing breadth 0.120 mm; halter length 0.180 mm. fore-femur: fore-tibia: fore-basi tarsi 15:12:8; mid-femur: mid-tibia: mid-basi tarsi 15:15:11; hind-femur: hind-tibia: hind-basi tarsi 20:24:13.

Mouth parts: Labrum is chitinized pointed structure with three apical sensilla. Mandible is with serrated teeth at the edge. Maxillary blade has visible teeth (17 laterals and 9 ventrals). Cibarium is with 40 teeth in a convex plate with a brown pigment patch. Pharynx is bottle shaped and with sharp teeth at base (Fig. 11b).


Fig. 10(a-b):	Female Sergentomyia (Grassomyia) dreyfussi turkestanica Theodor and Mesghali showing: (a) Cibarium, CT: Cibarial teeth and (b) Spermatheca (Sp)

Labrum length 0.110 mm; maxillary palp length 0.480 mm; palp formula (1,2,3,4,5) with a ratio of 3:7:10:11:33; proboscis length 0.120 mm; cibarial length 0.390 mm; third antennal segment (Aiii) 0.072 mm (Fig. 11a); Aiv 0.023 mm; Av 0.045 mm; antennal ascoid/sensilla chaetica on third antennal segment (Aaiii) 0.024 mm; Aaiv 0.050 mm; Aav 0.026 mm.

Wing venation: First radial vein (R₁) 0.500 mm; R₂ 0.270 mm; R₃ 0.540 mm; R₂₊₃ 0.600 mm; R₁ overlap 00 (No overlap).

Genitalia: Round egg shaped spermatheca (Fig. 11c). Tip with minute projections. The individual ducts are fused to form a short common duct and then fall in to the genital atrium. Spermatheca length 0.036 mm; breadth 0.020 mm.

Sergentomyia (Neophlebotomus) malayae (Lewis, 1973)
Specimen examined
Male: Not found in the collection.

Female: (4 specimens): Abdomen length 0.950-0.960 mm; abdomen breadth 0.220-0.225 mm; thorax length 0.480 mm; thorax breadth 0.510 mm; head length 0.420 mm; head breadth 0.300 mm; eye length 0.150 mm; eye breadth 0.110-0.120 mm; inter ocular 0.080 mm; wing length 0.150-0.160 mm; wing breadth 0.540 mm; halter length 0.190 mm. Legs are damaged.

Mouth parts: Labrum is chitinized. Maxillary blade has very fine teeth (2-3). Cibarium (Fig. 12c) is with a dark pigment patch. Convex arrangement of teeth is with larger medians and smaller laterals (12 teeth in total). Small teeth are found in three rows at the base. Pharynx armature is with scales and ridges at its broader base (Fig. 12a).

Labrum length 0.160-0.170 mm; maxillary palp length 0.880 mm; palp formula (1,2,3,4,5) or (1,(2,3,4),5) with a ratio of 4:13:14:15:27; proboscis length 0.180 mm; cibarial length 0.400 mm and width 0.580 mm; third antennal segment (Aiii) 0.280 mm; Aiv 0.170 mm; Av 0.165 mm; antennal ascoid/sensilla chaetica on third antennal segment (Aaiii) 0.041 mm; Aaiv 0.052 mm; Aav 0.050-0.052 mm.


Fig. 11(a-c):	Female Sergentomyia (Grassomyia) indica Theodor showing: (a) Antennal segments, Aiii: 3rd antennal segment, Aiv: 4th antennal segment, Av: 5th antennal segment, (b) Pharynx, PA: Pharyngeal armature and (c) Spermatheca (Sp)


Fig. 12(a-c):	Female Sergentomyia (Neophlebotomus) malayae A Kaul, Dhanda and Modi showing: (a) Pharynx, PA: Pharyngeal armature, (b) Spermatheca (Sp) and (c) Cibarium, CT: Cibarial teeth

Wing venation: First radial vein (R₁) 0.480 mm; R₂ 0.180 mm; R₃ 0.240-0.245 mm; R₂₊₃ 0.120 mm; R₁overlap 0.120 (large overlap).

Genitalia: Elliptical spermatheca (Fig. 12b) with striations. Spermatheca length 0.071 mm; breadth 0.030 mm.

DISCUSSION

The study sites include villages from Trincomalee, Anuradhapura and Vavuniya districts located in the Eastern, North Central and Northern provinces, respectively. There have been 15 CL cases reported from Trincomalee (Siripura village area) district from January 2009 to March 2010 and 10 cases in 2010 in Vavuniya (personal communication with Regional Health Offices). A total of 11 cases have been reported from Trincomalee district from June 2001 to June 2005 and 8 patients were recorded in the Anuradhapura district during the same period (Navaratna et al., 2007).

The present study and the report by Gajapathy and Surendran (2011) reveal the presence a rich species diversity of Sergentomyia sp. in CL endemic localities of the country. Three species of sandflies [Se. (Par.) franciscana, Se. (Par.) babu insularis (Lewis, 1978) and Se. (Par.) barraudi] belong to the subgenus Parrotomyia have already been reported from Sri Lanka (Ozbel et al., 2011). This is the first report for the presence of the following species in this subgenus; Se. (Par.) rudnicki, Se. (Par.) baghdadis, Se. (Par.) species A, Se. (Par.) modii, Se. (Par.) grekovi and Se. (Par.) bailyi. This is the first report for the presence of Sergentomyia (Neophlebotomus) malayae and Sergentomyia (Grassomyia) dreyfussi turkestanica from the country. The present article provides the first detailed morphological description of the sandfly species belong to these subgenera from Sri Lanka. Many previously undescribed taxonomic features were described in the text.

A recent report revealed the presence of L. donovani, the causative agent of CL in Sri Lanka, in field collected Phlebotomus argentipes s.l. in the country (Senanayake et al., 2011). However, as there are reports that support the vectorial role of Sergentomyia, there could be a possibility for the involvement of multi vector species in the transmission of leishmaniasis in Sri Lanka. Considering the difficulties in morphological based identification a reliable simple DNA based identification (Abaii et al., 2007; Hamarsheh et al., 2007a, b) assay is warranted with detailed morphometric analysis along with the seasonal distribution correlated with the disease cycle (Ibrahim et al., 2005), the abundance pattern (Aldawood et al., 2004; Al-Barrak, 2005) and the insecticide resistance level (Khoobdel, 2008) to study the epidemiology of the disease and to design an appropriate vector control measure in the country.

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