3. Loukrakpam (Two) revisi.cdr BIOTROPIA Vol. 21 No. 2, 2014: 91 - 98 TWO NEW FISCHER, 1894 (NEMATODA: TYLENCHINA) FROM MANIPUR, NORTHEAST INDIA Aphelenchoides LOUKRAKPAM BINA CHANU and NAOREM MOHILAL Received 25 February 2014/Accepted 1 October 2014 Two new nematode species of discovered from different localities of Manipur, North East India had been described in the present study. sp. nov. has a long stylet and four lateral lines which are gradually merged into two at the tail region. sp. nov. has a small body and short tail with a stylet length of 8.6 μm. sp. nov., sp. nov., Manipur, North- East India * Parasitology Section, Department of Life Sciences, Manipur University Canchipur - 795003, Manipur Aphelenchoides Aphelenchoides longistylus Aphelenchoides neominoris Aphelenchoides longistylus Aphelenchoides neominoris ABSTRACT INTRODUCTION Keywords: Aphelenchoides Aphelenchoides Aphelenchoides spp. are soil dwelling nematodes and often found in decaying plant materials. Their characteristic features include slender stylet with narrow lumen and usually with small basal knobs or swellings with presence of post uterine sac. Female tails are of medium length, conoid, with a tip pointed or rounded and often mucronate terminus. Most important characteristics of males include paired spicule, separate, rose thorn- shaped or derived from them with absence of gubernaculums. A number of spp. have been reported from India and from throughout the world. But description of new species of the genus is increasing due to intensive survey from different parts of the world including the biodiversity hot spot areas of North East India. The present manuscript deals with descriptions of two new species of spp. from Manipur, North East India. The discovery of the specimens is an added asset to the knowledge of the rich biodiversity of the region. * Corresponding author : bina.chanu@gmail.com DOI: 10.11598/btb.2014.21.2.3 93 MATERIALS AND METHODS RESULTS AND DISCUSSION Nematodes were extracted from collected soil samples by Baermann funnel technique. Collected nematodes were fixed in TAF (triethanolamine, formaldehyde and water) and processed by glycerol-ethanol method of Seinhorst (1959). Specimens were mounted in dehydrated glycerine. After slide preparation, measurements were taken using an ocular micrometer fixed on Nikon, Trinocular Research Microscope, model ECLIPSE E200 and diagrams were drawn under a drawing tube attached to the same microscope. sp. nov.Aphelenchoides longistylus 94 BIOTROPIA Vol. 21 No. 2, 2014 Table 1 Morphometric data of species of sp. nov.. Aphelenchoides longistylus Characters Holotype ♀ Paratypes ♀ s Paratype ♂s n 1 12 4 Stylet 24.22 24.22 24.22 Length 0.59 0.59 – 0.66 (0.625±27.79) 0.562 – 0.620 (0.59 ±0.02) a 34.4 34.4 - 35.85 (35.02±0.61) 38.2 – 42.02 (40.11±1.91) b 8.6 8.33 – 9.81(9.08±0.52) 7.24 –8.33 (7.78±0.54) b´ 6.03 6.03 – 6.49 (6.27±0.18) 3.25 – 5.603 (4.42±1.17) c 14.33 13.67 – 14.58 (14.19±0.38) 13.54 – 17.45 (15.22±1.43) c´ 6.00 5.6 – 6.14 (5.91±0.22) 4 – 6.2 (4.92±0.86) G1 40.69 39.39 – 43.08(41.05±1.52) - V 69.76 67.78 – 69.76 (69.08±0.92) - PVS/ V-A% 11.25 11.25 – 12.25(11.71±0.41) - T - - 92.61 – 102.84 (97.56±3.66) Max. body width 17.3 17.3 – 19.03(17.87±0.815) 13.38 Lip diameter 5.19 5.19 5.13 Lip height 1.73 1.73 1.73 Oesophagus 98.61 98.61 – 102.07 (99.76±1.63) - Median bulb length 13.84 13.84 13.84 – 15.57 (14.55±0.73) Median bulb diam. 10.38 8.65 - 10.38 (9.51±0.86) 8.65 –10.38 (9.53±0.70) Ovary length 242.2 242.2- 285.45 (257.35±19.88) - Testis - - 335.62 – 342.45 (339.14±1.91) Sperm theca 48.44 48.44 - PUS 15.57 8.65 – 18.65 (16.74±1.35) - PUS/VBD 0.9 0.9 – 1.07 (0.96±0.07) - Rectum 6.92 6.92 - Spicule - - 24.22 Tail 41.52 41.52 – 48.44 (44.16±3.05) 41.52 – 46.23 (43.89±1.91) ABD 6.92 6.92 – 8.65 (7.49±0.81) 10.32 Note: All measurements are in μm except Length is in mm Description Female: Male: Type habitat and locality: Type material: Differential diagnosis: Body is straight to curved ventrally upon fixation, 0.59 - 0.66 (0.62±27.79) mm in length and about 34.4 - 35.85 (35.02±0.61) times greatest body width long. Cuticle is finely annulated which is about 0.7 μm wide. Lateral fields with 4 longitu- dinal lines which merge into 2 lines at around tail regions. Cephalic region is indistinctly set off from body, appearing smooth and with 6 equal lips. Stylet is 24.22 μm long with indistinct basal knobs. Cylindrical portion of the stylet is slightly longer than anterior conical section. Procorpus is wider anteriorly, gradually narrowing at posterior part, 48.44 - 59.1 (52.48±4.15) μm long. Median bulb is spherical to pyriform in shape and occupying four-fifth of body width, 13.84 μm in length and 8.65 - 10.38 (9.51±0.86) μm in diameter. Oesophageal glands forms a non overlapping lobe with intestine, about 34.6 - 38.06 (36.60±1.28) μm in length. Nerve ring is about one-half of body width behind median bulb, encircling the anterior end of oesophageal glands. Excretory pore is at base of median bulb, 74.39 - 83.04 (78.33±3.22) μm long from anterior body region forming a lobe at dorsal side of nerve ring. Hemizonid, deirids and phasmids were not seen. Vulva is a transverse slit, two-fifth of body width long and is located approximately at one-half body length from anterior end. Genital tract is monoprodelphic, outstretched, usually extending up to the oesophageal gland lobe. Oocytes are arranged in a single row, post uterine sac about 2.15 - 2.31 (2.21±0.07) μm times anal body diameter long. Tail is 5.6 - 6.2 (5.9±0.22) μm times anal body diameter long, tapering gradually into a cylindrical tube terminating in a ventral prong tip. Found in abundance as is female, is slightly smaller than female and is more straight upon fixation. Cephalic region, stylet and oesophagus are as described for females. Tail is slender with a single terminal mucro. Spicules are about 24.22 μm long. Testis is single, 335.62 - 342.45 (339.14±2.51) μm long. Collected in December 2012 from soil around rhizospheric regions of coconut plant, Linn. at Ninghsing Khul, Jiri, Imphal West District, Manipur, India. Holotype female on the slide FSB - A -1/♀ sp. nov., paratype females on the slides FSB - A - 2- 12/♀ sp. nov., paratype males on the slides FSB - A - 1- 4/ ♂ sp. nov. and deposited in the Nematode collection center of Parasitology Section, Department of Life Sciences, Manipur University, Canchipur - 795003, Manipur, India. sp. nov. differs from all other species of Fischer, 1894 in possession of the longest stylet and 4 lateral lines which gradually merged into 2 at the tail regions. Cocos nucifera Aphelenchoides longistylus Aphelenchopides longistylus Aphelenchoides longistylus Aphelenchoides longistylus Aphelenchoides 3 3 3 95 Two new Fischer, 1894 (nematoda: tylenchina) Loukrakpam Bina ChanuAphelenchoides – et al. However, it comes close to (Hussain & Khan 1967); (Chawla & Khan 1979); (Tandon & Singh 1974) and (Hooper 1958). sp. nov. differs from (Hussain & Khan 1967) in having 4 lateral lines throughout body length, longer body length, larger values of a, b, c´, stylet and longer spicule; smaller values of c, V and indistinct stylet knobs (lateral lines =3, body length = 0.39 - 0.45, a= 30 - 33, b= 4 - 4.5, c´=3, stylet = 11 - 13, spicule = 15 - 19, c= 16 - 20, V = 69 - 77 and presence of stylet knobs in (Hussain & Khan 1967). sp. nov. comes close to (Chawla & Khan 1979) in having similar range of a, b, c and V values. But, it differs from (Chawla & Khan 1979) in having 4 lateral lines gradually merging into 2, larger values of body length, c value, stylet length and absence of stylet knobs (Lateral lines= 3, body length = 0.4 - 0.5 mm, stylet = 10 -11 μm and presence of stylet knobs in (Chawla & Khan 1979). sp. nov. also comes close to (Tandon & Singh 1974) in having similar range of body length, a, b, c values and spicule length. But, it differs from Tandon & Singh, 1974 in having larger value of c´, smaller value of V, longer stylet and 4 lateral lines which merged into 2 and indistinct stylet knobs (c´= 2.9, V = 71 - 73, stylet = 11 - 14 μm, 4 lateral lines and presence of stylet knobs in (Tandon & Singh 1974). Similarly, sp. nov. comes close to (Hooper 1958) in having similar range of body length, a, b, c and V values. But it differs from (Hooper 1958) in having 4 lateral lines, smaller values of b and longer stylet with indistinct stylet knobs and larger values of spicules (Lateral lines = 3, b= 10 - 14, stylet = 11 μm, spicules = 15 - 20 μm and presence of stylet knobs in (Hooper 1958). sp. nov. Body is finely annulated, tapering towards both extremities, straight and slightly curved ventrally on the tail region upon relaxation. Lateral fields have 4 incisures. Cephalic framework is smooth, set off from the body, about 5.19 μm wide and 2.53 μm high. Stylet is prominent with distinct stylet guards, 8.65 μm in length with distinct stylet knobs. Oesophagus has zig-zagly coiled procorpus, has strongly developed and rounded corpus with median sclerotised plates and elongated gland lobe overlying the intestine dorsally at a distance behind the bulb which is equal to about 3 - 4 times body width long. Neither deirids nor phasmids were seen. Excretory pore is close behind the nerve ring. Vulva is prominent, protruding both lips with vagina inclined. Large and elongated spermatheca, about 65.74 - 70.24 (67.99±2.25) μm long, oocytes are arranged in single row reaching up to the oesophageal bulb. Post-vulval uterine sac is about one-half vulval body width and is empty. Anterior lip of anus is protruding, tail is bluntly rounded, 12.11 - 25.95 (19.03±6.92) μm in length with a small hair like mucro. Aphelenchoides absari Aphelenchoides chalonus Aphelenchoides lanceolatus Aphelenchoides sacchari Aphelenchoides longistylus A. absari A. absari Aphelenchoides longistylus A. chalonus A. chalonus A. chalonus Aphelenchoides longistylus A. lanceolatus A. lanceolatus A. lanceolatus Aphelenchoides longistylus A. sacchari A. sacchari A. sacchari Aphelenchoides neominoris Description Female: 96 BIOTROPIA Vol. 21 No. 2, 2014 97 Male: Type habitat and locality: Type material: Differential diagnosis: Not found. Collected in September 2013 from soil around the rhizospheric regions of orange plant, from Sibilong, Chandel District, Manipur, India. Holotype female on the slide FSB-A - 2/ ♀ sp. nov., paratype females on the slides FSB - A - 1, 3 - 10/♀ sp. nov. and deposited at the Nematode collection center of Parasitology Section, Department of Life Sciences, Manipur University, Canchipur - 795003, Manipur, India. sp. nov. differs from all other species of Fischer, 1894 in having the shortest body length. But, it comes close to (Ebsary 1991); (Hussain & Khan 1967) and (Masleen 1979) in several other characters. 4 4 Aphelenchoides neominoris Aphelenchoides neominoris Aphelenchoides neominoris Aphelenchoides Aphelenchoides minoris Aphelenchoides absari Aphelenchoides vaughani Two new Fischer, 1894 (nematoda: tylenchina) Loukrakpam Bina ChanuAphelenchoides – et al. Characters Holotype Paratypes n 1 10 Stylet 8.65 8.65 Length 0.439 0.351 – 0.439 (383.48 ± 39.71) a 36.29 36.29 – 40.6 (38.44± 2.16) b 6.51 4.23 – 6.51 (5.37± 1. 14) b´ 6.03 6.03 – 6.49 (6.27±0.18) c 36.29 13.53 – 36.29 (24.90± 11.37) c´ 1.75 1.75 – 3.75 (2.75±1) G1 50 41.87 – 50.0 (45.93±4.06) V 71.65 69.95 – 71.65 (70.8±0.85) Lip width 5.19 5.19 Lip height 2.53 2.53 Oesophagus 67.47 67.47 – 83.04 (75.25±7.78) Median bulb width 8.65 6.92 – 8.63 (7.49±0.81) Ovary 219.71 147.05 – 219.71 (183.38±36.33) Spermatheca 65.74 65.74 – 70.24 (67.99±2.25) PUS 15.57 12.11 – 15.57 (13.84±1.73) PUS/VBD 1.28 1.28 – 1.4 (1.34±0.06) Rectum 5.19 5.19 Tail 12.11 12.11 – 25.95 (19.03±6.92) ABD 6.92 6.92 Table 2. Morphometric data of female species of sp. nov.Aphelenchoides neominoris Note: All measurements are in μm except Length is in mm 98 BIOTROPIA Vol. 21 No. 2, 2014 Legends to figures A - Female anterior body B - Male anterior body C - Female entire body D - Female reproductive system E - Female lateral lines F - Female tail region G - Male entire body H - Male tail region I - L. sp. nov. I - Female anterior body J - Female reproductive system K - Female lateral lines and L - Female tail region. Aphelenchoides neominoris Figure 1: A H. sp.nov.Aphelenchoides longistylus 99 Aphelenchoides neominoris Aphelenchoides minoris Aphelenchoides minoris A. minoris Aphelenchoides neominoris A. absari A. absari A. absari Aphelenchoides neominoris Aphelenchoides vaughani A. vaughani A. vaughani NO. SB/FT/LS-113/2013 sp. nov. comes close to (Ebsary 1991) in having smaller body length and a ventral mucro. But, it differs from (Ebsary 1991) in having 4 lateral lines, larger values of a, c; smaller values of b, PUS, stylet and indistinct stylet knobs (lateral lines = 3, a= 26 - 29, b= 8- 9, c= 15, PUS= 5 VBD, stylet= 10 μm and presence of stylet knobs in (Ebsary 1991). sp. nov. comes close to (Hussain & Khan 1967) in having almost similar body length, 4 lateral lines and presence of ventral mucro. But, it differs from Hussain & Khan, 1967 in having larger values of a, b and c but smaller values of c´, stylet length, presence of post uterine sac and indistinct stylet knobs (a= 30 - 33, b= 4.0 - 4.5, c= 16 - 20, c´=3, stylet = 11- 13 μm, absence of post uterine sac and presence of stylet knobs in (Hussain & Khan 1967). sp. nov. is similar to (Masleen 1979) in having almost similar body length, presence of 4 lateral lines and ventral mucros. But, the species differs from (Masleen 1979) in having larger values of a, c and G and smaller values of b, stylet length, tail length and in having a simple ventral mucro (a= 24- 31, b= 6.3 - 9.2, c= 12.1 - 16.9, G = 32 - 44, stylet = 9.5 - 11 μm, tail = 23.5 - 38.5 μm with a ventral mucro minutely multi-papillate at its tip in (Masleen 1979). The authors thanked the Head, Department of Life Sciences, Manipur University, Canchipur for providing necessary laboratory facilities. The corresponding author also acknowledged the Science and Engineering Research Board, Department of Science & Technology, Govt. of India for financial support in the form of Fast Track Young Scientist fellowship (SERB ). 1; 1 ACKNOWLEDGEMENTS REFERENCES AEbsary BA. 1991. . Canada: Agriculture Canada. IV. Chawla ML, Khan E.1979. Two nomenclatorial corrections. Indian J Nematol 7: 100. Fisher M. 1894. Über eine Clematis -krankheit. Bericht aus dem Physiolischen Laboratorium des Landwirthschaftlichen, Instituts der Universitat Halle (11)3: 1 - 11. Hooper DJ. 1958. n. sp. and n. sp. (Nematoda: Aphelenchoidea). Nematologica 3: 228 - 35. Hussain SI, Khan AM.1967. On the status of the genera of the superfamily Aphelenchoidea (Fuchs, 1937). Proc Helm Soc Wash 34: 167 - 74. Maslen NR. 1979. Six new nematode species from maritime Antarctic. Nematologica 25: 288 - 308. Tandon RS, Singh SP. 1974. Nematode parasites of the common sponge gourd, from Lucknow Geobios 1: 24 - 7. Seinhorst JW.1959. A rapid method for the transfer of nematodes from fixative to anhydrous glycerine. Nematologica 4: 67 - 9. Catalog of the Order Tylenchida (Nematoda) Aphelenchoides dactylocercus A. sacchari Luffa cylindfrica . Two new Fischer, 1894 (nematoda: tylenchina) Loukrakpam Bina ChanuAphelenchoides – et al.